Differential progression of neonatal diethylstilbestrol-induced disruption of the hamster testis and seminal vesicle.
The synthetic estrogen diethylstilbestrol (DES) is now recognized as the prototypical endocrine disruptor. Using a hamster experimental system, we performed a detailed temporal assessment of how neonatal DES-induced disruption progresses in the testis compared to the seminal vesicle. Both morphological and Western blot analyses confirmed that neonatal DES exposure alters androgen responsiveness in the male hamster reproductive tract. We also determined that the disruption phenomenon in the male hamster is manifest much earlier in the seminal vesicle than in the testis and that testis disruption often occurs differently between the pair of organs in a given animal. In the neonatally DES-exposed seminal vesicle, histopathological effects included: (1) general atrophy, (2) lack of exocrine products, (3) epithelial dysplasia, (4) altered organization of stromal cells and extracellular matrix, and (5) striking infiltration with polymorphonuclear leukocytes. Also, the morphological disruption phenomenon in the seminal vesicle was accompanied by a range of up-regulation and down-regulation responses in the whole organ levels of various proteins.


A theory of frequency domain invariants: spherical harmonic identities for BRDF/lighting transfer and image consistency.
This paper develops a theory of frequency domain invariants in computer vision. We derive novel identities using spherical harmonics, which are the angular frequency domain analog to common spatial domain invariants such as reflectance ratios. These invariants are derived from the spherical harmonic convolution framework for reflection from a curved surface. Our identities apply in a number of canonical cases, including single and multiple images of objects under the same and different lighting conditions. One important case we consider is two different glossy objects in two different lighting environments. For this case, we derive a novel identity, independent of the specific lighting configurations or BRDFs, that allows us to directly estimate the fourth image if the other three are available. The identity can also be used as an invariant to detecttampering in the images. While this paper is primarily theoretical, it has the potential to lay the mathematical foundations for two important practical applications. First, we can develop more general algorithms for inverse rendering problems, which can directly relight and change material properties by transferring the BRDF or lighting from another object or illumination. Second, we can check the consistency of an image, to detect tampering or image splicing.


The three-dimensional structure of Cys-47-modified mouse liver glutathione S-transferase P1-1. Carboxymethylation dramatically decreases the affinity for glutathione and is associated with a loss of electron density in the alphaB-310B region.
The three-dimensional structure of mouse liver glutathione S-transferase P1-1 carboxymethylated at Cys-47 and its complex with S-(p-nitrobenzyl)glutathione have been determined by x-ray diffraction analysis. The structure of the modified enzyme described here is the first structural report for a Pi class glutathione S-transferase with no glutathione, glutathione S-conjugate, or inhibitor bound. It shows that part of the active site area, which includes helix alphaB and helix 310B, is disordered. However, the environment of Tyr-7, an essential residue for the catalytic reaction, remains unchanged. The position of the sulfur atom of glutathione is occupied in the ligand-free enzyme by a water molecule that is at H-bond distance from Tyr-7. We do not find any structural evidence for a tyrosinate form, and therefore our results suggest that Tyr-7 is not acting as a general base abstracting the proton from the thiol group of glutathione. The binding of the inhibitor S-(p-nitrobenzyl)-glutathione to the carboxymethylated enzyme results in a partial restructuring of the disordered area. The modification of Cys-47 sterically hinders structural organization of this region, and although it does not prevent glutathione binding, it significantly reduces the affinity. A detailed kinetic study of the modified enzyme indicates that the carboxymethylation increases the Km for glutathione by 3 orders of magnitude, although the enzyme can function efficiently under saturating conditions.


Prevalence of Trauma-Induced Hypocalcemia in the Prehospital Setting.
Recent data published by the Special Operations community suggest the Lethal Triad of Trauma should be changed to the Lethal Diamond, to include coagulopathy, acidosis, hypothermia, and hypocalcemia. The purpose of this study is to determine the prevalence of trauma-induced hypocalcemia in level I and II trauma patients. This is a retrospective cohort study conducted at a level I trauma center and Special Operations Combat Medic (SOCM) training site. Adult patients were identified via trauma services registry from September 2021 to April 2022. Patients who received blood products prior to emergency department (ED) arrival were excluded from the study. Ionized calcium levels were utilized in this study. Of the 408 patients screened, 370 were included in the final analysis of this cohort. Hypocalcemia was noted in 189 (51%) patients, with severe hypocalcemia identified in two (<1%) patients. Thirty-two (11.2%) patients had elevated international normalized ratio (INR), 34 (23%) patients had pH <7.36, 21 (8%) patients had elevated lactic acid, and 9 (2.5%) patients had a temperature of <35°C. Hypocalcemia was prevalent in half of the trauma patients in this cohort. The administration of a calcium supplement empirically in trauma patients from the prehospital environment and prior to blood transfusion is not recommended until further data prove it beneficial.


Adaptive Symptom Monitoring Using Hidden Markov Models - An Application in Ecological Momentary Assessment.
Wearable and mobile technology provides new opportunities to manage health conditions remotely and unobtrusively. For example, healthcare providers can repeatedly sample a person's condition to monitor progression of symptoms and intervene if necessary. There is usually a utility-tolerability trade-off between collecting information at sufficient frequencies and quantities to be useful, and over-burdening the user or the underlying technology, particularly when active input is required from the user. Selecting the next sampling time adaptively using previous responses, so that people are only sampled at high frequency when necessary, can help to manage this trade-off. We present a novel approach to adaptive sampling using clustered continuous-time hidden Markov models. The model predicts, at any given sampling time, the probability of moving to an 'alert' state, and the next sample time is scheduled when this probability has exceeded a given threshold. The clusters, each representing a distinct sub-model, allow heterogeneity in states and state transitions. The work is illustrated using longitudinal mental-health symptom data in 49 people collected using ClinTouch, a mobile app designed to monitor people with a diagnosis of schizophrenia. Using these data, we show how the adaptive sampling scheme behaves under different model parameters and risk thresholds, and how the average sampling can be substantially reduced whilst maintaining a high sampling frequency during high-risk periods.


Impact of early life exposures to geohelminth infections on the development of vaccine immunity, allergic sensitization, and allergic inflammatory diseases in children living in tropical Ecuador: the ECUAVIDA birth cohort study.
Geohelminth infections are highly prevalent infectious diseases of childhood in many regions of the Tropics, and are associated with significant morbidity especially among pre-school and school-age children. There is growing concern that geohelminth infections, particularly exposures occurring during early life in utero through maternal infections or during infancy, may affect vaccine immunogenicity in populations among whom these infections are endemic. Further, the low prevalence of allergic disease in the rural Tropics has been attributed to the immune modulatory effects of these infections and there is concern that widespread use of anthelmintic treatment in high-risk groups may be associated with an increase in the prevalence of allergic diseases. Because the most widely used vaccines are administered during the first year of life and the antecedents of allergic disease are considered to occur in early childhood, the present study has been designed to investigate the impact of early exposures to geohelminths on the development of protective immunity to vaccines, allergic sensitization, and allergic disease. A cohort of 2,403 neonates followed up to 8 years of age. Primary exposures are infections with geohelminth parasites during the last trimester of pregnancy and the first 2 years of life. Primary study outcomes are the development of protective immunity to common childhood vaccines (i.e. rotavirus, Haemophilus influenzae type B, Hepatitis B, tetanus toxoid, and oral poliovirus type 3) during the first 5 years of life, the development of eczema by 3 years of age, the development of allergen skin test reactivity at 5 years of age, and the development of asthma at 5 and 8 years of age. Potential immunological mechanisms by which geohelminth infections may affect the study outcomes will be investigated also. The study will provide information on the potential effects of early exposures to geohelminths (during pregnancy and the first 2 years of life) on the development of vaccine immunity and allergy. The data will inform an ongoing debate of potential effects of geohelminths on child health and will contribute to policy decisions on new interventions designed to improve vaccine immunogenicity and protect against the development of allergic diseases. Current Controlled Trials ISRCTN41239086.


The preferred traits of mates in a cross-national study of heterosexual and homosexual men and women: an examination of biological and cultural influences.
BBC Internet survey participants (119,733 men and 98,462 women) chose from a list of 23 traits those they considered first, second, and third most important in a relationship partner. Across all participants, the traits ranked most important were: intelligence, humor, honesty, kindness, overall good looks, face attractiveness, values, communication skills, and dependability. On average, men ranked good looks and facial attractiveness more important than women did (d = 0.55 and 0.36, respectively), whereas women ranked honesty, humor, kindness, and dependability more important than men did (ds = 0.23, 0.22, 0.18, and 0.15). Sexual orientation differences were smaller than sex differences in trait rankings, but some were meaningful; for example, heterosexual more than homosexual participants assigned importance to religion, fondness for children, and parenting abilities. Multidimensional scaling analyses showed that trait preference profiles clustered by participant sex, not by sexual orientation, and by sex more than by nationality. Sex-by-nation ANOVAs of individuals' trait rankings showed that sex differences in rankings of attractiveness, but not of character traits, were extremely consistent across 53 nations and that nation main effects and sex-by-nation interactions were stronger for character traits than for physical attractiveness. United Nations indices of gender equality correlated, across nations, with men's and women's rankings of character traits but not with their rankings of physical attractiveness. These results suggest that cultural factors had a relatively greater impact on men's and women's rankings of character traits, whereas biological factors had a relatively greater impact on men's and women's rankings of physical attractiveness.


The effect of delivery mechanisms on the uptake of bed net re-impregnation in Kilifi District, Kenya.
The results of recently completed trials in Africa of insecticide-treated bed nets (ITBN) offer new possibilities for malaria control. These experimental trials aimed for high ITBN coverage combined with high re-treatment rates. Whilst necessary to understand protective efficacy, the approaches used to deliver the intervention provide few indications of what coverage of net re-treatment would be under operational conditions. Varied delivery and financing strategies have been proposed for the sustainable delivery of ITBNs and re-treatment programmes. Following the completion of a randomized, controlled trial on the Kenyan coast, a series of suitable delivery strategies were used to continue net re-treatment in the area. The trial adopted a bi-annual, house-to-house re-treatment schedule free of charge using research project staff and resulted in over 95% coverage of nets issued to children. During the year following the trial, sentinel dipping stations were situated throughout the community and household members informed of their position and opening times. This free re-treatment service achieved between 61-67% coverage of nets used by children for three years. In 1997 a social marketing approach, that introduced cost-retrieval, was used to deliver the net re-treatment services. The immediate result of this transition was that significantly fewer of the mothers who had used the previous re-treatment services adopted this revised approach and coverage declined to 7%. The future of new delivery services and their financing are discussed in the context of their likely impact upon previously defined protective efficacy and cost-effectiveness estimates.


Progression-Free and Overall Survival in ALK-Positive NSCLC Patients Treated with Sequential Crizotinib and Ceritinib.
Anaplastic lymphoma kinase (ALK) rearrangements are important therapeutic targets in non-small cell lung cancer (NSCLC) that confer sensitivity to the ALK inhibitors crizotinib and ceritinib. To determine the outcome of sequential treatment with crizotinb and ceritinib, we retrospectively evaluated a cohort of ALK-positive patients treated with both agents. We identified 73 ALK-positive NSCLC patients treated with crizotinib followed by ceritinib at four institutions. Medical records were reviewed to determine overall survival (OS) and progression-free survival (PFS) on crizotinib and ceritinib. Among 73 ALK-positive patients, the median PFS (mPFS) on crizotinib was 8.2 months [95% confidence interval (CI), 7.4-10.6]. The median interval from crizotinib discontinuation to initiation of ceritinib was 25 days (range, 1-694). The mPFS on ceritinib was 7.8 months (6.5-9.1). Among 53 patients with no interval therapies between crizotinib and ceritinib, the mPFS on ceritinib was similar at 7.8 months (5.4-9.8). The median combined PFS for sequential treatment with crizotinib and ceritinib was 17.4 months (15.5-19.4). Among 23 patients who underwent post-crizotinib/pre-ceritinib biopsies, there was no difference in PFS on ceritinib between patients with or without ALK resistance mutations (mPFS 5.8 vs. 6.5 months, respectively; P = 0.510). In the overall study population, median OS was 49.4 months (35.5-63.1). Ceritinib has significant antitumor activity in ALK-positive NSCLC-even when crizotinib immediately precedes treatment with ceritinib (median combined PFS 17.0 months). Additional studies are necessary to further define the impact of specific ALK resistance mutations on duration of response to ceritinib.


Effects of a long-acting LHRH agonist preparation on plasma gonadotropin and prolactin levels in castrated male rats and on the release of prolactin from ectopic pituitaries.
We have examined the effects of a single subcutaneous injection of an LHRH agonist, D-Trp-6-LHRH, in biodegradable microcapsules of poly(DL-lactide-co-glycolide) on plasma gonadotropin and prolactin (PRL) levels in castrated and in castrated-hypophysectomized-pituitary grafted (CAST-APX-GRAFT) male rats. The results were compared to the effects of daily injections of the same LHRH agonist dissolved in saline. In castrated rats, there were no significant alterations in plasma LH or PRL levels during the 10 days following the injection of LHRH agonist microcapsules, while FSH levels were generally reduced. In castrated males given daily injections of 6 micrograms of LHRH agonist in saline, plasma LH levels were significantly reduced while plasma PRL levels were not changed. In CAST-APX-GRAFT rats, both D-Trp-6-LHRH microcapsules and daily LHRH agonist injections appeared to increase plasma PRL levels. The pattern of changes in PRL release in both groups was similar, with levels on day 6 being significantly higher than those measured on days 1, 3 and 10 after onset of treatment. As expected, LH and FSH levels in these animals were extremely low. Immunoreactive D-Trp-6-LHRH was consistently detectable in the plasma of CAST-APX-GRAFT animals after microcapsule administration, whereas in animals given daily injections of this agonist in saline, its plasma concentrations were often below the detectability limit of the employed assay. These findings suggest that the LHRH agonist, D-Trp-6-LHRH, is capable of causing a short term stimulation of PRL release from ectopic pituitaries. Elevation of plasma LH levels is apparently not required for this effect.


Electrical stimulation of the left and right human fusiform gyrus causes different effects in conscious face perception.
Neuroimaging and electrophysiological studies across species have confirmed bilateral face-selective responses in the ventral temporal cortex (VTC) and prosopagnosia is reported in patients with lesions in the VTC including the fusiform gyrus (FG). As imaging and electrophysiological studies provide correlative evidence, and brain lesions often comprise both white and gray matter structures beyond the FG, we designed the current study to explore the link between face-related electrophysiological responses in the FG and the causal effects of electrical stimulation of the left or right FG in face perception. We used a combination of electrocorticography (ECoG) and electrical brain stimulation (EBS) in 10 human subjects implanted with intracranial electrodes in either the left (5 participants, 30 FG sites) or right (5 participants, 26 FG sites) hemispheres. We identified FG sites with face-selective ECoG responses, and recorded perceptual reports during EBS of these sites. In line with existing literature, face-selective ECoG responses were present in both left and right FG sites. However, when the same sites were stimulated, we observed a striking difference between hemispheres. Only EBS of the right FG caused changes in the conscious perception of faces, whereas EBS of strongly face-selective regions in the left FG produced non-face-related visual changes, such as phosphenes. This study examines the relationship between correlative versus causal nature of ECoG and EBS, respectively, and provides important insight into the differential roles of the right versus left FG in conscious face perception.


Extensive citrus triploid hybrid production by 2x×4x sexual hybridizations and parent-effect on the length of the juvenile phase.
The citrus fresh market demands the production of seedless citrus fruits, as seedy fruits are not accepted by consumers. The recovery of triploid plants has proven to be the most promising approach to achieve this goal, since triploids have very low fertility, are generally seedless and do not induce seeds in other cultivars by cross pollination. Triploid plants can be recovered by 2x×4x sexual hybridization. In this work, we present an effective methodology to recover triploid plants from 2x×4x hybridizations based on in vitro embryo rescue, ploidy level analysis by flow cytometry and genetic origin of triploid plants. The pollen viability of diploid and tetraploid citrus genotypes was analyzed by comparing the pollen germination rate in vitro. The pollen viability of tetraploid (doubled-diploid) genotypes is generally reduced but sufficient for successful pollination. Triploid embryos were identified in normal and undeveloped seeds that did not germinate under greenhouse conditions. The influence of parents and environmental conditions on obtaining triploid plants was analyzed and a strong interaction was noted between the parents and environmental conditions. The parental effect on the length of the juvenile phase was also demonstrated through observations of a large number of progeny over the last 15 years. The juvenile phase length of the triploid hybrids obtained with 'Fortune' mandarin as female parent and tetraploid 'Orlando' tangelo as male parent was shorter than the juvenile phase obtained with a clementine as female parent and tetraploids of 'Nova', 'W. Leaf' and 'Pineapple' male parents. Effective methodology to recover citrus triploid plants from 2x×4x sexual hybridizations and the parental effect on the length of the juvenile phase.


Many human accelerated regions are developmental enhancers.
The genetic changes underlying the dramatic differences in form and function between humans and other primates are largely unknown, although it is clear that gene regulatory changes play an important role. To identify regulatory sequences with potentially human-specific functions, we and others used comparative genomics to find non-coding regions conserved across mammals that have acquired many sequence changes in humans since divergence from chimpanzees. These regions are good candidates for performing human-specific regulatory functions. Here, we analysed the DNA sequence, evolutionary history, histone modifications, chromatin state and transcription factor (TF) binding sites of a combined set of 2649 non-coding human accelerated regions (ncHARs) and predicted that at least 30% of them function as developmental enhancers. We prioritized the predicted ncHAR enhancers using analysis of TF binding site gain and loss, along with the functional annotations and expression patterns of nearby genes. We then tested both the human and chimpanzee sequence for 29 ncHARs in transgenic mice, and found 24 novel developmental enhancers active in both species, 17 of which had very consistent patterns of activity in specific embryonic tissues. Of these ncHAR enhancers, five drove expression patterns suggestive of different activity for the human and chimpanzee sequence at embryonic day 11.5. The changes to human non-coding DNA in these ncHAR enhancers may modify the complex patterns of gene expression necessary for proper development in a human-specific manner and are thus promising candidates for understanding the genetic basis of human-specific biology.


Protocol for exercise hemodynamic assessment: performing an invasive cardiopulmonary exercise test in clinical practice.
Invasive cardiopulmonary exercise testing (iCPET) combines full central hemodynamic assessment with continuous measurements of pulmonary gas exchange and ventilation to help in understanding the pathophysiology underpinning unexplained exertional intolerance. There is increasing evidence to support the use of iCPET as a key methodology for diagnosing heart failure with preserved ejection fraction and exercise-induced pulmonary hypertension as occult causes of exercise limitation, but there is little information available outlining the methodology to use this diagnostic test in clinical practice. To bridge this knowledge gap, the operational protocol for iCPET at our institution is discussed in detail. In turn, a standardized iCPET protocol may provide a common framework to describe the evolving understanding of mechanism(s) that limit exercise capacity and to facilitate research efforts to define novel treatments in these patients.


Monocyte Subsets and Related Chemokines in Carotid Artery Stenosis and Ischemic Stroke.
Carotid stenosis (CS) is an important cause of ischemic stroke. However, reliable markers for the purpose of identification of high-risk, so-called vulnerable carotid plaques, are still lacking. Monocyte subsets are crucial players in atherosclerosis and might also contribute to plaque rupture. In this study we, therefore, aimed to investigate the potential role of monocyte subsets and associated chemokines as clinical biomarkers for vulnerability of CS. Patients with symptomatic and asymptomatic CS (n = 21), patients with cardioembolic ischemic strokes (n = 11), and controls without any cardiovascular disorder (n = 11) were examined. Cardiovascular risk was quantified using the Essen Stroke Risk Score (ESRS). Monocyte subsets in peripheral blood were measured by quantitative flow cytometry. Plaque specimens were histologically analyzed. Furthermore, plasma levels of monocyte chemotactic protein 1 (MCP-1) and fractalkine were measured. Intermediate monocytes (Mon2) were significantly elevated in symptomatic and asymptomatic CS-patients compared to controls. Mon2 counts positively correlated with the ESRS. Moreover, stroke patients showed an elevation of Mon2 compared to controls, independent of the ESRS. MCP-1 levels were significantly higher in patients with symptomatic than in those with asymptomatic CS. Several histological criteria significantly differed between symptomatic and asymptomatic plaques. However, there was no association of monocyte subsets or chemokines with histological features of plaque vulnerability. Due to the multifactorial influence on monocyte subsets, the usability as clinical markers for plaque vulnerability seems to be limited. However, monocyte subsets may be critically involved in the pathology of CS.


The impact of IAIMS at Georgetown: strategies and outcomes.
Integration of multiple information systems of a medical center will change the way physicians work and practice medicine in the future. Several major steps must be taken by an institution to make this a reality. Since 1983, Georgetown has been engaged in an Integrated Academic Information Management System (IAIMS) project to bring together multiple sources of information that reside on different computers and database systems. Georgetown is developing a Biotechnology and Biomedical Knowledge Network that includes informational and clinical databases, scholar workstations, instruction on computer use, a campuswide network with local area network nodes, and a modular approach to systems integration. The IAIMS project, spearheaded by the medical library, has enabled a broad spectrum of health professionals to benefit directly from new, dynamic information services. The network is heavily used; in 1991, more than 2,100 individual users conducted more than 148,500 computer functions and more than 104,000 searches. There is economy of scale in high-volume use. Overall, the average search cost is $1.57; for high use databases the cost is $0.38, and for low use, it is $9.41. As described in this paper, IAIMS offers a cost-effective means of enhancing patient care by improving information services to physicians. At Georgetown, IAIMS has advanced the concept of integration, accelerated use of computers in education, increased user acceptance of advanced technologies, and established cost factors for providing information resources. While progress made in improving the transfer of medical information is impressive, it is clear that IAIMS requires several more years of support to achieve full implementation.


Magnetic resonance imaging features of empty sella in dogs.
In humans, the empty sella is defined as herniation of the subarachnoidal space into the sella turcica with invisible (total empty sella) or reduced (partial empty sella) hypophyseal size. An empty sella can be associated with endocrine disturbances. The purpose of this study was to determine if the empty sella exists in dogs or cats and whether it is associated with endocrinopathy or hydrocephalus. Archived magnetic resonance images of the head of 370 dogs and 77 cats were re-evaluated specifically for the presence or absence of the pituitary gland. Hypophyseal tissue filling more than 50% of the sella was considered normal, between 30% and 50% was defined as partial empty sella and less than 30% was defined as total empty sella. In patients with an empty sella, and in all other imaged dogs of the affected breeds, the ventricle to brain ratio was measured. In patients with an empty sella, the medical record was searched for evidence of endocrinopathy. No cat had an empty sella but 11 dogs (3%) had a small or missing hypophysis. Seven of these dogs had a ventricle to brain ratio greater than 15%. Only small dogs (mean weight of 6.7kg) were affected. One dog with an empty sella had signs of central hyperadrenocorticism while the other 10 had no sign of endocrinopathy. We conclude that the empty sella exists in dogs but is not generally associated with endocrinopathy.


Genetic epidemiology of dengue viruses in phase III trials of the CYD tetravalent dengue vaccine and implications for efficacy.
This study defined the genetic epidemiology of dengue viruses (DENV) in two pivotal phase III trials of the tetravalent dengue vaccine, CYD-TDV, and thereby enabled virus genotype-specific estimates of vaccine efficacy (VE). Envelope gene sequences (n = 661) from 11 DENV genotypes in 10 endemic countries provided a contemporaneous global snapshot of DENV population genetics and revealed high amino acid identity between the E genes of vaccine strains and wild-type viruses from trial participants, including at epitope sites targeted by virus neutralising human monoclonal antibodies. Post-hoc analysis of all CYD14/15 trial participants revealed a statistically significant genotype-level VE association within DENV-4, where efficacy was lowest against genotype I. In subgroup analysis of trial participants age 9-16 years, VE estimates appeared more balanced within each serotype, suggesting that genotype-level heterogeneity may be limited in older children. Post-licensure surveillance is needed to monitor vaccine performance against the backdrop of DENV sequence diversity and evolution.


Are trajectories of self-regulation abilities from ages 2-3 to 6-7 associated with academic achievement in the early school years?
The aim of this study was to estimate the association between two key aspects of self-regulation, 'task attentiveness' and 'emotional regulation' assessed from ages 2-3 to 6-7 years, and academic achievement when children were aged 6-7 years. Participants (n = 3410) were children in the Longitudinal Study of Australian Children. Parents rated children's task attentiveness and emotional regulation abilities when children were aged 2-3, 4-5 and 6-7. Academic achievement was assessed using the Academic Rating Scale completed by teachers. Linear regression models were used to estimate the association between developmental trajectories (i.e. rate of change per year) of task attentiveness and emotional regulation, and academic achievement at 6-7 years. Improvements in task attentiveness between 2-3 and 6-7 years, adjusted for baseline levels of task attentiveness, child and family confounders, and children's receptive vocabulary and non-verbal reasoning skills at age 6-7 were associated with greater teacher-rated literacy [B = 0.05, 95% confidence interval (CI) = 0.04-0.06] and maths achievement (B = 0.04, 95% CI = 0.03-0.06) at 6-7 years. Improvements in emotional regulation, adjusting for baseline levels and covariates, were also associated with better teacher-rated literacy (B = 0.02, 95% CI = 0.01-0.04) but not with maths achievement (B = 0.01, 95% CI = -0.01-0.02) at 6-7 years. For literacy, improvements in task attentiveness had a stronger association with achievement at 6-7 years than improvements in emotional regulation. Our study shows that improved trajectories of task attentiveness from ages 2-3 to 6-7 years are associated with improved literacy and maths achievement during the early school years. Trajectories of improving emotional regulation showed smaller effects on academic outcomes. Results suggest that interventions that improve task attentiveness when children are aged 2-3 to 6-7 years have the potential to improve literacy and maths achievement during the early school years.


CCL25/CCR9 interactions regulate the function of iNKT cells in oxazolone-induced colitis in mice.
Natural killer T (NKT) cells share phenotypic and functional properties with both conventional natural killer cells and T cells. These cells might have an important role in the pathogenesis of ulcerative colitis (UC). The interaction of chemokine ligand 25 (CCL25) with chemokine receptor 9 (CCR9) is involved in gut-specific migration of leukocytes and induces regulatory T cells (Tregs) to migrate to the intestine in chronic ileitis. In UC patients, NKT receptor CD161, CCL25, and CCR9 expression levels were evaluated by qRT-PCR. A murine model of oxazolone-induced colitis was induced in BALB/c mice. The mRNA levels of NK1.1, CCL25 and CCR9, and pro-inflammatory cytokines in mice were evaluated. The CCR9 expression on Type I or invariant NKT (iNKT) cells, and the iNKT cells chemotaxis are observed according to flow cytometry. NKT receptor CD161, CCL25 and CCR9 expression levels were significantly increased in UC patients. And, the mRNA expression levels of NK1.1, CCL25 and CCR9 were increased in oxazolone-induced colitis in mice. The production of pro-inflammatory cytokines was significantly increased, especially interleukin 4 (IL-4), IL-10 and IL-13. We observed significantly increased CCR9 expression on iNKT cells. Furthermore, we found an increased iNKT population and enhanced chemotaxis during oxazolone-induced colitis. Our study suggests that CCL25/CCR9 interactions may promote the induction and function of iNKT cells during oxazolone-induced colitis. These findings may have important implications for UC treatment and suggest a role for CCR9 inhibitors.


An epidemiologic survey of shoulder pain in chiropractic practice in australia.
This survey investigated the demographic characteristics of the responding practitioners, the prevalence of shoulder pain symptoms seen in weekly chiropractic practice, the sources of shoulder pain, the chiropractor-diagnosed prevalence of shoulder clinical syndromes, and the management strategies for Australian chiropractors. A survey was created by the authors consisting of questions seeking demographic information from the respondents, shoulder syndrome, and shoulder management information. The survey was mailed to every chiropractic practitioner based in the Australian state of New South Wales (general population 6.8 million in 2005). Contact details were derived from Yellow Pages online listings. One thousand thirty-seven surveys were mailed to New South Wales-based chiropractors, with 192 (21%) returning a completed survey. The prevalence of shoulder pain symptoms as reported by the practitioners was 12% of the total weekly patients, with the major cause of symptoms related to overuse (32%). The most prevalent working diagnosis of shoulder pain was shoulder impingement syndrome (13%), followed by impingement syndrome with rotator cuff tendinosis (17%), impingement syndrome without rotator cuff tendinosis (14%), and chiropractic shoulder subluxation (12%). Shoulder pain is managed with a combination of manipulation, mainly diversified technique (81%), peripheral joint manipulation (82%), and various soft tissue strategies used by 92% of practitioners. Rehabilitation strategies were also used by 89% of practitioners with a main emphasis placed on rotator cuff strengthening. The results suggest a moderate prevalence of shoulder pain in clinical practice with the most prevalent structure involved being the rotator cuff tendon. Most practitioners use a multimodal therapeutic treatment approach in managing disorders of the shoulder.


Novel therapeutic strategy for neurodegeneration by blocking Aβ seeding mediated aggregation in models of Alzheimer's disease.
Aβ accumulation plays a central role in the pathogenesis of Alzheimer's disease (AD). Recent studies suggest that the process of Aβ nucleated polymerization is essential for Aβ fibril formation, pathology spreading and toxicity. Therefore, targeting this process represents an effective therapeutic strategy to slow or block disease progression. To discover compounds that might interfere with the Aβ seeding capacity, toxicity and pathology spreading, we screened a focused library of FDA-approved drugs in vitro using a seeding polymerization assay and identified small molecule inhibitors that specifically interfered with Aβ seeding-mediated fibril growth and toxicity. Mitoxantrone, bithionol and hexachlorophene were found to be the strongest inhibitors of fibril growth and protected primary cortical neuronal cultures against Aβ-induced toxicity. Next, we assessed the effects of these three inhibitors in vivo in the mThy1-APPtg mouse model of AD (8-month-old mice). We found that mitoxantrone and bithionol, but not hexachlorophene, stabilized diffuse amyloid plaques, reduced the levels of Aβ42 oligomers and ameliorated synapse loss, neuronal damage and astrogliosis. Together, our findings suggest that targeting fibril growth and Aβ seeding capacity constitutes a viable and effective strategy for protecting against neurodegeneration and disease progression in AD.


Consensus statement from the International Consensus Meeting on the Role of Decompressive Craniectomy in the Management of Traumatic Brain Injury : Consensus statement.
Two randomised trials assessing the effectiveness of decompressive craniectomy (DC) following traumatic brain injury (TBI) were published in recent years: DECRA in 2011 and RESCUEicp in 2016. As the results have generated debate amongst clinicians and researchers working in the field of TBI worldwide, it was felt necessary to provide general guidance on the use of DC following TBI and identify areas of ongoing uncertainty via a consensus-based approach. The International Consensus Meeting on the Role of Decompressive Craniectomy in the Management of Traumatic Brain Injury took place in Cambridge, UK, on the 28th and 29th September 2017. The meeting was jointly organised by the World Federation of Neurosurgical Societies (WFNS), AO/Global Neuro and the NIHR Global Health Research Group on Neurotrauma. Discussions and voting were organised around six pre-specified themes: (1) primary DC for mass lesions, (2) secondary DC for intracranial hypertension, (3) peri-operative care, (4) surgical technique, (5) cranial reconstruction and (6) DC in low- and middle-income countries. The invited participants discussed existing published evidence and proposed consensus statements. Statements required an agreement threshold of more than 70% by blinded voting for approval. In this manuscript, we present the final consensus-based recommendations. We have also identified areas of uncertainty, where further research is required, including the role of primary DC, the role of hinge craniotomy and the optimal timing and material for skull reconstruction.


Editorial Commentary: Arthroscopic Outcomes Are Equal to Open Popliteus Tendon Reconstructions, but Do Not Forget That the Goal Is a Stable Posterior Cruciate Ligament Reconstruction.
Arthroscopic popliteus tendon reconstructions were found to be equally as efficacious as open popliteus reconstructions in restoring external rotation stability. However, when the procedure was combined with a posterior cruciate ligament (PCL) reconstruction, the PCL reconstruction still had increases of 4 to 5 mm of posterior tibial translation in side-to-side differences on posterior stress radiographs. Thus, the goal of restoring stability for combined popliteus tendon and PCL reconstructions is still elusive.


Quantification of the amount of dental material removed by selective grinding in wax dentures with photogrammetric measurements.
The use of photogrammetry may be a new method to quantify the amount of artificial dental material removed from the surface of each teeth during the grind procedure (SG). SG is necessary in each denture to reach a correct occlusion. It consists in a refine action on the prosthesis teeth's surface using milling machine tools, aimed to remove the interferences (pre-contacts) between upper and lower teeth during chewing. This measure is achieved after a comparison between pre and post-grinding 3D models. This new application could be of interest for both dentists and dental technicians because it could be used to evaluate, with a accurate numerical description, the action applied on teeth surfaces during the grinding process. Furthermore, results of the analysis could have some value for the dental industry, since the use of photogrammetry can improve the process, reducing costs during the design of artificial teeth and eventually this method could be used as a teaching tool both for dental and "dental technician" high school students. The purpose of this work is to measure the thickness of the artificial enamel removed during grinding phases. Usually, the dental technician adjusts the dental plate on the mount of the patient following the traditional method, without a quantitative evaluation of the material removed. The photogrammetric method (PM) proposed here allows to measure the amount of material removed during the grinding process. This measure is achieved after a comparison between pre and post-grinding 3D models. Under control of three teachers (experts of dentures performed according to the Gerber method) ten complete dentures arrangements (upper and inferior arches) performed by dental students at the Prosthodontic Department of the University of Parma, Italy were analyzed with PM before and after SG. The average thickness variation between the pre and post-grinding dentures is within the range of 0.1÷0.4 mm. For the upper arches, the mean value of the SG process is 223 µm while for the inferior arches is 240 µm. Results show that the most important grind process in all models appear in correspondence of cusps, with values up to 1660 µm. On the other hand, in correspondence of the fossae the results show a moderate grind action: the value is around 200-300 µm. Conversely to guidelines thought to students: cusps undergo a greater grinding process than fossae, consequently cusps should be revisioned at least on their technical and morphological aspects. The average thickness variation between the pre and post-grinding dentures is within the range of 0.1÷0.4 mm, this mean an equal value loss of vertical dimension. Furthermore, the knowledge of the gauge material removed during the SG could be useful for dental industries, giving important information, that could be considered for project and design of artificial teeth. The FM implemented in this article has given satisfactory preliminary results, showing good accuracy, low costs and high versatility. It is necessary to highlight that this is an experimental method and that the present analysis is a pilot study that needs further evaluation. Nevertheless results obtained could be of some value for medical companies, in order to improve the artificial teeth's design and project. Moreover, such a method may serve as educational tool for dental students.


Economic support to improve tuberculosis treatment outcomes in South Africa: a pragmatic cluster-randomized controlled trial.
Poverty undermines adherence to tuberculosis treatment. Economic support may both encourage and enable patients to complete treatment. In South Africa, which carries a high burden of tuberculosis, such support may improve the currently poor outcomes of patients on tuberculosis treatment. The aim of this study was to test the feasibility and effectiveness of delivering economic support to patients with pulmonary tuberculosis in a high-burden province of South Africa. This was a pragmatic, unblinded, two-arm cluster-randomized controlled trial, where 20 public sector clinics acted as clusters. Patients with pulmonary tuberculosis in intervention clinics (n = 2,107) were offered a monthly voucher of ZAR120.00 (approximately US$15) until the completion of their treatment. Vouchers were redeemed at local shops for foodstuffs. Patients in control clinics (n = 1,984) received usual tuberculosis care. Intention to treat analysis showed a small but non-significant improvement in treatment success rates in intervention clinics (intervention 76.2%; control 70.7%; risk difference 5.6% (95% confidence interval: -1.2%, 12.3%), P = 0.107). Low fidelity to the intervention meant that 36.2% of eligible patients did not receive a voucher at all, 32.3% received a voucher for between one and three months and 31.5% received a voucher for four to eight months of treatment. There was a strong dose-response relationship between frequency of receipt of the voucher and treatment success (P <0.001). Our pragmatic trial has shown that, in the real world setting of public sector clinics in South Africa, economic support to patients with tuberculosis does not significantly improve outcomes on treatment. However, the low fidelity to the delivery of our voucher meant that a third of eligible patients did not receive it. Among patients in intervention clinics who received the voucher at least once, treatment success rates were significantly improved. Further operational research is needed to explore how best to ensure the consistent and appropriate delivery of such support to those eligible to receive it.


Analyzing Single Molecule Localization Microscopy Data Using Cubic Splines.
The resolution of super-resolution microscopy based on single molecule localization is in part determined by the accuracy of the localization algorithm. In most published approaches to date this localization is done by fitting an analytical function that approximates the point spread function (PSF) of the microscope. However, particularly for localization in 3D, analytical functions such as a Gaussian, which are computationally inexpensive, may not accurately capture the PSF shape leading to reduced fitting accuracy. On the other hand, analytical functions that can accurately capture the PSF shape, such as those based on pupil functions, can be computationally expensive. Here we investigate the use of cubic splines as an alternative fitting approach. We demonstrate that cubic splines can capture the shape of any PSF with high accuracy and that they can be used for fitting the PSF with only a 2-3x increase in computation time as compared to Gaussian fitting. We provide an open-source software package that measures the PSF of any microscope and uses the measured PSF to perform 3D single molecule localization microscopy analysis with reasonable accuracy and speed.


Perigestational high folic acid: impact on offspring's peripheral metabolic response.
Offspring of dams exposed to excess folic acid during the perigestational period have been shown by us to be predisposed to metabolic dysfunction revealed by hyperglycemia, glucose intolerance, increased insulin and decreased adiponectin in late adulthood. This work aims to characterize adipocyte phenotype and expression profile of genes in the regulation of lipid and glucose metabolism in visceral adipose tissue and in skeletal muscle. From mating until weaning, a recommended dose of folic acid for pregnancy (C, 2 mg of folic acid per kg of diet) or a high folic acid dose (HFA, 40 mg of folic acid per kg of diet) was administered to Sprague-Dawley females. At 10 months of age progeny were divided into groups fed the standard chow (C/STD and HFA/STD) and groups fed the standard chow plus drinking water with 10% fructose (C/FRU and HFA/FRU), as an additional metabolic challenge. Adipocyte morphology and quantification of key genes involved in lipid and glucose metabolism were studied in visceral adipose tissue and skeletal muscle of 13 months old offspring. HFA exposure led to an enlargement of visceral adipose cells most likely mediated by an upregulation of lipoprotein lipase, and it tended to downregulate Glut4 in visceral adipose tissue and skeletal muscle. Fructose exposure in a background of perigestational excess folic acid, but not in controls, induced an upregulation of lipogenesis pathway genes and it decreased jejunal expression of the proton-coupled folate transporter (Pcft1). In addition, fructose exposure led to a downregulation of jejunal Sglt1 in control animals. Our data suggest that high folic acid exposure during the perigestational period caused morphologic and genic alterations related to insulin resistant states indicating that this intervention may act as an effective programmer of long-term metabolic dysfunction.


Traditional risk factors of acute coronary syndrome in four different male populations - total cholesterol value does not seem to be relevant risk factor.
Cardiovascular diseases are the most common cause of mortality and morbidity in most populations. As the traditional modifiable risk factors (smoking, hypertension, dyslipidemia, diabetes mellitus, and obesity) were defined decades ago, we decided to analyze recent data in patients who survived acute coronary syndrome (ACS). The Czech part of the study included data from 999 males, and compared them with the post-MONICA study (1,259 males, representing general population). The Lithuanian study included 479 male patients and 456 age-matched controls. The Kazakhstan part included 232 patients and 413 controls. In two countries, the most robust ACS risk factor was smoking (OR 3.85 in the Czech study and 5.76 in the Lithuanian study), followed by diabetes (OR 2.26 and 2.07) and hypertension (moderate risk elevation with OR 1.43 and 1.49). These factors did not influence the ACS risk in Kazakhstan. BMI had no significant effect on ACS and plasma cholesterol was surprisingly significantly lower (P<0.001) in patients than in controls in all countries (4.80+/-1.11 vs. 5.76+/-1.06 mmol/l in Czechs; 5.32+/-1.32 vs. 5.71+/-1.08 mmol/l in Lithuanians; 4.88+/-1.05 vs. 5.38+/-1.13 mmol/l in Kazakhs/Russians). Results from our study indicate substantial heterogeneity regarding major CVD risk factors in different populations with the exception of plasma total cholesterol which was inversely associated with ACS risk in all involved groups. These data reflect ethnical and geographical differences as well as changing pattern of cardiovascular risk profiles.


Adsorption of polyvinyl alcohol from wastewater by sintered porous red mud.
Several types of red mud-based porous materials (RMPM) and other raw minerals via different processes were prepared and characterized using X-ray diffraction (XRD) analyses and scanning electron microscope (SEM) observations. Using the polymer sponge method, a 72% apparent porosity could be reached compared with 64% by adding a pore-forming agent. These materials were tested for their adsorption of polyvinyl alcohol (PVA) from simulated textile wastewater. The best mass ratio of RMPM to PVA solution was 50:100 with a removal maximum of 25.8% after they were in contact for 50 min. The adsorption rate and kinetics could be better described by Lagergren's pseudo-second-order model in comparison with the pseudo-first-order model.


Neuron-interacting satellite glial cells in human trigeminal ganglia have an APC phenotype.
Satellite glial cells (SGC) in sensory ganglia tightly envelop the neuronal cell body to form discrete anatomical units. This type of glial cell is considered neuroectoderm-derived and provides physical support to neuron somata. There are scattered hints in the literature suggesting that SGC have an immune-related function within sensory ganglia. In this study, we addressed the hypothesis that SGC are tissue-resident APC. The immune phenotype and function of a large series (n = 40) of human trigeminal ganglia (TG) were assessed by detailed flow cytometry, in situ analyses, and functional in vitro assays. Human TG-resident SGC (TG-SGC) uniformly expressed the common leukocyte marker CD45, albeit at lower levels compared with infiltrating T cells, and the macrophage markers CD14, CD68, and CD11b. In addition, TG-SGC expressed the myeloid dendritic cell (DC) marker CD11c, the T cell costimulatory molecules CD40, CD54, CD80, and CD86 and MHC class II. However, the mature DC marker CD83 was absent on TG-SGC. Functionally, TG-SGC phagocytosed fluorescent bacteria, but were unable to induce an allogeneic MLR. Finally, TG-infiltrating T cells expressed the T cell inhibitory molecules CD94/NKG2A and PD-1, and the interacting TG-SGC expressed the cognate ligands HLA-E and PD-L1, respectively. In conclusion, the data demonstrate that human TG-SGC have a unique leukocyte phenotype, with features of both macrophages and immature myeloid DC, indicating that they have a role as TG-resident APC with potential T cell modulatory properties.


Chimpanzees' responses to the dead body of a 9-year-old group member.
The social behavior of chimpanzees has been extensively studied, yet not much is known about how they behave in response to the death of a group member. Here, we provide a detailed report of the reactions of a group of chimpanzees to finding the dead body of a 9-year-old male group member. The behavior of the group was characterized by quiet attendance and close inspections punctuated by rare displays. Moreover, the body was continuously attended and closely inspected by several adults and juveniles, including an adult male who formed a close social bond with the deceased individual after the deceased individual's mother died 4 years earlier. When considered with observations of how chimpanzees respond to dead infants and adults in this group and in others, these observations suggest that chimpanzees' responses to death may be mediated by social bonds with the deceased individual. The results are discussed in light of recent reports on chimpanzees' reactions to dead community members and more general primate thanatology. Am. J. Primatol. 78:914-922, 2016. © 2016 Wiley Periodicals, Inc.


Microtubule-dependent transport of secretory vesicles visualized in real time with a GFP-tagged secretory protein.
Biosynthetic transport from the trans-Golgi network (TGN) to the plasma membrane (PM) is mediated by secretory vesicles. We analyzed secretory vesicle transport in real time using a GFP-tagged secretory protein, hCgB-GFP, consisting of human chromogranin B (hCgB) and green fluorescent protein (GFP). The fusion protein was expressed transiently in Vero cells or in a stable clone after induction with butyrate. After arrest of the biosynthetic protein transport at 20 degrees C, fluorescent hCgB-GFP colocalized with TGN38, a marker of the TGN. Subsequent release of the secretion block at 37 degrees C led to the formation of green fluorescent vesicles. Confocal analysis revealed that these vesicles were devoid of TGN38 and of Texas Red-coupled transferrin and cathepsin D, markers of the endosomal/lysosomal pathway. As determined by fluorometry and metabolic labelling hCgB-GFP was secreted from the TGN to the PM with a t(1/2) of 20-30 minutes. Video-microscope analysis of green fluorescent vesicles showed brief periods of rapid directed movement with maximal velocities of 1 microm/second. Vesicle movement occurred in all directions, centrifugal, centripetal and circumferential, and 50% of the vesicles analyzed reversed their direction of movement at least once within an observation period of 45 seconds. In the presence of nocodazole the movement of fluorescent vesicles ceased. Concomitantly, secretion of hCgB-GFP was slowed but not completely blocked. We suggest that microtubules (MT) facilitate the delivery of secretory vesicles to the PM by a stochastic transport, thereby increasing the probability for a vesicle/target membrane encounter.


Isolated growth hormone (GH) deficiency in adult patients: baseline clinical characteristics and responses to GH replacement in comparison with hypopituitary patients. A sub-analysis of the KIMS database.
Isolated growth hormone deficiency (IGHD) provides the ideal model to characterize GHD without interference from other pituitary deficiencies or their treatment. No study has addressed the question whether adult patients with IGHD differ in clinical presentation or in responsiveness to GH replacement from adult patients with multiple pituitary hormone deficiencies (MPHD) receiving conventional replacement therapy. Data were retrieved from the outcomes research database KIMS (Pfizer international metabolic database). Patients with IGHD accounted for 9.6% (274/2868) of all GHD patients. Patients were separated according to the timing of onset. In the adult-onset (AO) group, 167 patients with IGHD were compared to 1992 patients with MPHD. In the childhood-onset (CO) group, 107 patients with IGHD were compared to 602 patients with MPHD. To assess the effect of GH replacement after one year, a longitudinal sub-analysis in the AO group was performed comparing 89 IGHD patients to 1234 MPHD patients. The same study was done in the CO group comparing 66 IGHD patients to 386 MPHD patients. Because IGHD patients were significantly younger than MPHD patients, data analysis was also performed after adjustment for gender and age. In the AO group, non-functioning and secreting pituitary adenomas were the most common primary diagnoses in both IGHD and MPHD. Medical history revealed a high prevalence of hypertension and fractures in both subgroups, but also of non-insulin dependent diabetes mellitus. The prevalence of obesity was high and the waist circumference was elevated. The lipid profile was unfavourable in both IGHD and MPHD. IGF-I concentration and SDS were comparable in both subgroup. Quality of life assessed by QoL-AGHDA was equally poor in both IGHD and MPHD. GH replacement therapy induced favourable changes without distinction. In the CO group, the most common cause in both subgroups was idiopathic. Fracture rate was similarly prevalent in both IGHD and MPHD. Obesity was prominent in both subgroups, but BMI and waist circumference were lower in IGHD. Adverse lipid changes were similarly found in both IGHD and MPHD. IGF-I concentration and SDS were significantly higher in the IGHD subgroup compared to the MPHD subgroup. The QoL-AGHDA score was equally abnormal in both IGHD and MPHD. GH replacement achieved similar significant improvement in both subgroups. GHD patients with AO-IGHD and AO-MPHD present with a similar clinical expression and respond similarly to GH replacement. Patients with CO-IGHD are less severely affected by GHD than CO-MPHD patients, but, nevertheless, both groups show a comparable adverse lipid profile and poor quality of life and respond favourably to GH replacement. These findings support the concept that GH alone is responsible for most if not all metabolic aspects of hypopituitary patients receiving conventional replacement therapy, regardless of age of onset or aetiology. As a consequence, GH replacement therapy not only has potential benefit in GHD patients with additional hormonal deficits, but also the indication of treatment must be extended to patients with isolated GHD.


An intestinal Trojan horse for gene delivery.
The intestinal epithelium forms an essential element of the mucosal barrier and plays a critical role in the pathophysiological response to different enteric disorders and diseases. As a major enteric dysfunction of the intestinal tract, inflammatory bowel disease is a genetic disease which results from the inappropriate and exaggerated mucosal immune response to the normal constituents in the mucosal microbiota environment. An intestine targeted drug delivery system has unique advantages in the treatment of inflammatory bowel disease. As a new concept in drug delivery, the Trojan horse system with the synergy of nanotechnology and host cells can achieve better therapeutic efficacy in specific diseases. Here, we demonstrated the feasibility of encapsulating DNA-functionalized gold nanoparticles into primary isolated intestinal stem cells to form an intestinal Trojan horse for gene regulation therapy of inflammatory bowel disease. This proof-of-concept intestinal Trojan horse will have a wide variety of applications in the diagnosis and therapy of enteric disorders and diseases.


Haplotype tagging for the identification of common disease genes.
Genome-wide linkage disequilibrium (LD) mapping of common disease genes could be more powerful than linkage analysis if the appropriate density of polymorphic markers were known and if the genotyping effort and cost of producing such an LD map could be reduced. Although different metrics that measure the extent of LD have been evaluated, even the most recent studies have not placed significant emphasis on the most informative and cost-effective method of LD mapping-that based on haplotypes. We have scanned 135 kb of DNA from nine genes, genotyped 122 single-nucleotide polymorphisms (SNPs; approximately 184,000 genotypes) and determined the common haplotypes in a minimum of 384 European individuals for each gene. Here we show how knowledge of the common haplotypes and the SNPs that tag them can be used to (i) explain the often complex patterns of LD between adjacent markers, (ii) reduce genotyping significantly (in this case from 122 to 34 SNPs), (iii) scan the common variation of a gene sensitively and comprehensively and (iv) provide key fine-mapping data within regions of strong LD. Our results also indicate that, at least for the genes studied here, the current version of dbSNP would have been of limited utility for LD mapping because many common haplotypes could not be defined. A directed re-sequencing effort of the approximately 10% of the genome in or near genes in the major ethnic groups would aid the systematic evaluation of the common variant model of common disease.


Combination of selenium and green tea improves the efficacy of chemoprevention in a rat colorectal cancer model by modulating genetic and epigenetic biomarkers.
Dietary supplementation of selenium and green tea holds promise in cancer prevention. In this study, we evaluated the efficacies of selenium and green tea administered individually and in combination against colorectal cancer in an azoxymethane (AOM)-induced rat colonic carcinogenesis model and determined the underlying mechanisms of the protection. Four-week old Sprague-Dawley male rats were fed with diets containing 0.5% green tea extract, 1 ppm selenium as selenium-enriched milk protein, or combination of 1 ppm selenium and 0.5% green tea extract. Animals received 2 AOM (15 mg/kg) treatments to induce colonic oncogenesis. Rats were killed 8 or 30 wk later after the last AOM to examine the effect of dietary intervention on aberrant crypt foci (ACF) formation or tumor development. On sacrifice, colons were examined for ACF and tumors, the mRNA levels of SFRP5 and Cyclin D1, and the proteins levels of ß-catenin, COX-2, Ki-67, DNMT1 and acetyl histone H3. The combination of selenium and green tea resulted in a significant additive inhibition of large ACF formation, this effect was greater than either selenium or green tea alone, P<0.01; the combination also had a significant additive inhibition effect on all tumor endpoints, the effect of the combination diet on tumor incidence, multiplicity and size was greater than selenium or green tea alone, P<0.01. Rats fed the combination diet showed marked reduction of DNMT1 expression and induction of histone H3 acetylation, which were accompanied by restoration of SFRP5 mRNA in normal-appearing colonic crypts. The combination diet also significantly reduced ß-catenin nuclear translocation, Cyclin D1 expression and cell proliferation. These data show, for the first time, that combination of selenium and green tea is more effective in suppressing colorectal oncogenesis than either agent alone. The preventive effect is associated with regulation of genetic and epigenetic biomarkers implicated in colonic carcinogenesis.


Three-dimensional quantitative analysis of healthy foot shape: a proof of concept study.
Foot morphology has received increasing attention from both biomechanics researches and footwear manufacturers. Usually, the morphology of the foot is quantified by 2D footprints. However, footprint quantification ignores the foot's vertical dimension and hence, does not allow accurate quantification of complex 3D foot shape. The shape variation of healthy 3D feet in a population of 31 adult women and 31 adult men who live in Belgium was studied using geometric morphometric methods. The effect of different factors such as sex, age, shoe size, frequency of sport activity, Body Mass Index (BMI), foot asymmetry, and foot loading on foot shape was investigated. Correlation between these factors and foot shape was examined using multivariate linear regression. The complex nature of a foot's 3D shape leads to high variability in healthy populations. After normalizing for scale, the major axes of variation in foot morphology are (in order of decreasing variance): arch height, combined ball width and inter-toe distance, global foot width, hallux bone orientation (valgus-varus), foot type (e.g. Egyptian, Greek), and midfoot width. These first six modes of variation capture 92.59% of the total shape variation. Higher BMI results in increased ankle width, Achilles tendon width, heel width and a thicker forefoot along the dorsoplantar axis. Age was found to be associated with heel width, Achilles tendon width, toe height and hallux orientation. A bigger shoe size was found to be associated with a narrow Achilles tendon, a hallux varus, a narrow heel, heel expansion along the posterior direction, and a lower arch compared to smaller shoe size. Sex was found to be associated with differences in ankle width, Achilles tendon width, and heel width. Frequency of sport activity was associated with Achilles tendon width and toe height. A detailed analysis of the 3D foot shape, allowed by geometric morphometrics, provides insights in foot variations in three dimensions that can not be obtained from 2D footprints. These insights could be applied in various scientific disciplines, including orthotics and shoe design.


Microbial nar-GFP cell sensors reveal oxygen limitations in highly agitated and aerated laboratory-scale fermentors.
Small-scale microbial fermentations are often assumed to be homogeneous, and oxygen limitation due to inadequate micromixing is often overlooked as a potential problem. To assess the relative degree of micromixing, and hence propensity for oxygen limitation, a new cellular oxygen sensor has been developed. The oxygen responsive E. coli nitrate reductase (nar) promoter was used to construct an oxygen reporter plasmid (pNar-GFPuv) which allows cell-based reporting of oxygen limitation. Because there are greater than 109 cells in a fermentor, one can outfit a vessel with more than 109 sensors. Our concept was tested in high density, lab-scale (5 L), fed-batch, E. coli fermentations operated with varied mixing efficiency - one verses four impellers. In both cases, bioreactors were maintained identically at greater than 80% dissolved oxygen (DO) during batch phase and at approximately 20% DO during fed-batch phase. Trends for glucose consumption, biomass and DO showed nearly identical behavior. However, fermentations with only one impeller showed significantly higher GFPuv expression than those with four, indicating a higher degree of fluid segregation sufficient for cellular oxygen deprivation. As the characteristic time for GFPuv expression (approx 90 min.) is much larger than that for mixing (approx 10 s), increased specific fluorescence represents an averaged effect of oxygen limitation over time and by natural extension, over space. Thus, the pNar-GFPuv plasmid enabled bioreactor-wide oxygen sensing in that bacterial cells served as individual recirculating sensors integrating their responses over space and time. We envision cell-based oxygen sensors may find utility in a wide variety of bioprocessing applications.


Effects of monetary contingencies on smoking relapse: influences of trait depression, personality, and habitual nicotine intake.
Of 56 male smokers, 34 were randomly assigned (by 60% random odds) to quit smoking immediately, whereas the remaining 22 were assigned to quit after an additional 31 days. Compensation ($300) was contingent on abstinence for a minimum of 31 or 2 days (depending on random assignment) and completion of all experimental sessions. Contingencies for the immediate-quit group required 31 days of abstinence; those for the delayed-quit group required only 2 days of abstinence. Contingency duration (31 vs. 2 days) predicted days to relapse. All but 4 of the 31-day contingency participants maintained abstinence for at least 31 days, whereas only 3 of the 2-day contingency group abstained for 31+ days. However, 31-day contingencies did not result in longer postcontingency time to relapse. Higher trait neuroticism, depression, and psychopathic deviate scores predicted decreased time to relapse. Prequit cotinine concentrations and Fagerström Tolerance Questionnaire scores failed to predict time to relapse.


A deep crust-mantle boundary in the asteroid 4 Vesta.
The asteroid 4 Vesta was recently found to have two large impact craters near its south pole, exposing subsurface material. Modelling suggested that surface material in the northern hemisphere of Vesta came from a depth of about 20 kilometres, whereas the exposed southern material comes from a depth of 60 to 100 kilometres. Large amounts of olivine from the mantle were not seen, suggesting that the outer 100 kilometres or so is mainly igneous crust. Here we analyse the data on Vesta and conclude that the crust-mantle boundary (or Moho) is deeper than 80 kilometres.


Inhibitory effects of Opuntia humifusa on 7, 12-dimethyl- benz[a]anthracene and 12-O-tetradecanoylphorbol-13- acetate induced two-stage skin carcinogenesis.
Opuntia humifusa, member of the Cactaceae family, was previously demonstrated to have radical scavenging, anti-inflammatory and anti-proliferative effects in in vitro models. It was suggested that O. humifusa could function in the prevention of carcinogenesis. To investigate the in vivo chemopreventive effect of O. humifusa, mice were fed a diet containing either 1% or 3% following 7, 12-dimethylbenz[a] anthracene (DMBA) and 12-O-tetradecanoylphorbol-13-acetate (TPA) induction of skin carcinogenesis. Significant decrease in the numbers of papilloma and epidermal hyperplasia were observed in mice fed with O. humifusa, compared to the control group. O. humifusa also upregulated high total antioxidant capacity and level of phase II detoxifying enzyme such as superoxide dismutase and glutathione S-transferase activity in the skin. Lipid peroxidation activity level was measured in skin cytosol and significantly inhibited in 3% OH fed group compared to the control group. These results suggest that O. humifusa exerts chemopreventive effects on chemical carcinogenesis in mouse skin and that prevention effects are associated with reduction of oxidative stress via the modulation of cutaneous lipid peroxidation, enhancing of total antioxidant capacity especially in phase II detoxifying enzyme system and partial apoptotic influence.


Dynamics and reliability of bistable neurons driven with time-dependent stimuli.
The reliability of a spiking neuron depends on the frequency content of the driving input signal. Previous studies have shown that well above threshold, regularly firing neurons generate reliable responses when the input signal resonates with the firing frequency of the cell. Instead, well below threshold, reliable responses are obtained when the input frequency resonates with the subthreshold oscillations of the neuron. Previous theories, however, provide no clear prediction for the input frequency giving rise to maximally reliable spiking at threshold, which is probably the most relevant firing regime in mammalian cortex under physiological conditions. In particular, when the firing onset is governed by a subcritical Hopf bifurcation, the frequency of subthreshold oscillations often differs from the firing rate at threshold. The predictions of previous studies, hence, cannot be smoothly merged at threshold. Here we explore the behavior of reliability in bistable neurons near threshold using three types of driving stimuli: constant, periodic, and stochastic. We find that the two natural frequencies of the system, associated with the two coexisting attractors, provide a rich variety of possible locking modes with the external signal. Reliability is determined by the sensitivity to noise of each locking mode and by the transition probabilities between modes. Noise increases the amount of spike time jitter, and minimal jitter is obtained for input frequencies coinciding with the suprathreshold firing rate of the cell. In addition, noise may either enhance or inhibit transitions between the two attractors, depending on the input frequency. The dual role played by noise in bistable systems implies that reliability is determined by a delicate balance between spike time jitter and the rate of transitions between attractors.


Reconstruction of burn scar of the upper extremities with artificial skin.
The management of upper-extremity burn contractures is a major challenge for plastic surgeons. After approval by the Food and Drug Administration, artificial skin (Integra) has been available in Taiwan since 1997. From January of 1997 to July of 1999, the authors applied artificial skin to 13 severely burned patients for the reconstruction of their upper extremities, resulting in an increased range of motion in the upper-extremity joints and improved skin quality. An additional benefit was the rapid reepithelialization of the donor sites. There were no complications of infection throughout the therapeutic course, and the overall results were satisfactory. During the 2-year study, scar condition was monitored between 8 and 24 months, and a good appearance and pliable skin were obtained according to the Vancouver Scar Scale. According to this evaluation of Oriental skin turgor, normal pigmentation was restored about 6 months after the resurfacing procedure. For patients with severe burns in whom there is insufficient available skin for a full-thickness skin graft or another appropriate flap for scar revision, Integra is an alternative. The two major concerns in dealing with artificial skin are (1) a 10- to 14-day waiting period for maturation of the neo-dermis, necessitating a two-stage operation, and (2) prevention of infection with antibiotics and meticulous wound care.


Relationship between tail lesions and lung health in slaughter pigs.
Tail lesions are associated with poor health either because they serve as a point of entry for pathogens or because of shared risk factors. This study investigated the relationship between carcass tail lesion and lung lesion severity scores in slaughter pigs. Carcasses were scored after scalding/dehairing for tail lesion severity (0-4). Lungs were scored according to an adapted version of the BPEX pig health scheme. Severity of enzootic pneumonia (EP-like lesions) was recorded on a scale of 0-50. Severity of pleurisy was scored on a 0-2 scale with score 2 equating to severe pleurisy or those lungs that remained attached to the chest wall ('lungs in chest'). The database for assessing pleurisy lesions contained all pleurisy scores (n=5628). Lungs with a score of 2 for pleurisy were excluded from the analysis of all other lung lesions as such lungs could not be assessed for other lesions (n=4491). Associations between tail lesions and different lung lesion outcomes were analysed using generalized linear mixed models (PROC GLIMMIX) with random effect for batch. Males were more affected by moderate (OR=1.9, 95% CI 1.51-2.34) and severe (OR=5.8, 95% CI 3.45-9.70) tail lesions than females. EP-like lesions and pleurisy were most commonly observed. Pigs with severe tail lesions tended to have more 'lungs in chest' than pigs with moderate tail lesions (P=0.1). No other associations between tail lesions and lung lesions were found. Males had higher odds of having EP-like lesions (OR=1.2, 95% CI 1.05-1.36) than females. Tail lesions on the carcass may not be an accurate predictor of lung health. However, tail lesions are important welfare indicators and respiratory disease is a significant infectious condition affecting pigs. Thus, recording of tail and lung lesions at meat inspection provides valuable information regarding on-farm health and welfare of pigs.


Use of a phage display library to identify oligopeptides binding to the lumenal surface of polarized endothelium by ex vivo perfusion of human umbilical veins.
Human endothelial-specific targeting peptides were identified by biopanning within freshly-obtained human umbilical cords. Umbilical veins were cleaned in situ and M13 phage display libraries were passed through the cords. Tightly bound phage were recovered following isolation of endothelial cells by collagenase digestion and homogenisation, allowing production of enriched phage libraries for subsequent rounds of panning. After five rounds of biopanning, five promising sequences were selected and the binding of the corresponding phage clones was compared in perfused umbilical veins. Each of these peptides showed substantial binding, although the clone encoding the heptapeptide KPSGLTY showed the greatest, some 89-times greater than insertless phage. Binding of this phage clone was examined to cells in vitro, where it demonstrated at least five-times greater binding to isolated human umbilical vein endothelial cells than to 911, SKOV3, B16F10 and Cos7 cells. These initial peptides may prove useful targeting agents for endothelial-selective delivery, and this powerful approach should be readily applicable to biopanning in a broad range of human vessels ex vivo.


Upregulated ATM gene expression and activated DNA crosslink-induced damage response checkpoint in Fanconi anemia: implications for carcinogenesis.
Fanconi anemia (FA) predisposes to hematopoietic failure, birth defects, leukemia, and squamous cell carcinoma of the head and neck (HNSCC) and cervix. The FA/BRCA pathway includes 8 members of a core complex and 5 downstream gene products closely linked with BRCA1 or BRCA2. Precancerous lesions are believed to trigger the DNA damage response (DDR), and we focused on the DDR in FA and its putative role as a checkpoint barrier to cancer. In primary fibroblasts with mutations in the core complex FANCA protein, we discovered that basal expression and phosphorylation of ATM (ataxia telangiectasia mutated) and p53 induced by irradiation (IR) or mitomycin C (MMC) were upregulated. This heightened response appeared to be due to increased basal levels of ATM in cultured FANCA-mutant cells, highlighting the new observation that ATM can be regulated at the transcriptional level in addition to its well-established activation by autophosphorylation. Functional analysis of this response using gamma-H2AX foci as markers of DNA double-stranded breaks (DSBs) demonstrated abnormal persistence of only MMC- and not IR-induced foci. Thus, we describe a processing defect that leads to general DDR upregulation but specific persistence of DNA crosslinker-induced damage response foci. Underscoring the significance of these findings, we found resistance to DNA crosslinker-induced cell cycle arrest and apoptosis in a TP53-mutant, patient-derived HNSCC cell line, whereas a lymphoblastoid cell line derived from this same individual was not mutated at TP53 and retained DNA crosslinker sensitivity. Our results suggest that cancer in FA may arise from selection for cells that escape from a chronically activated DDR checkpoint.


Identification of cyclic intermediates in Azorhizobium caulinodans nicotinate catabolism.
In wild-type Azorhizobium caulinodans ORS571, nicotinate served both as anabolic substrate for NAD+ production and as catabolic substrate for use as the N source. Catabolic enzyme activities were greatest from cultures grown with nicotinate as the N source and least when cultures were grown with ammonium as the N source. Vector insertion mutants unable to catabolize nicotinate (nic::Vi mutants) still required micromolar quantities of this compound for growth. Therefore, A. caulinodans wild type is NAD+ auxotrophic. As the first two intermediates in A. caulinodans nicotinate catabolism, two cyclic compounds, 6-hydroxynicotinate and 1,4,5,6-tetrahydro-6-oxonicotinate, were identified. These compounds were purified from the growth medium of strain 61009 (a nic::Vi mutant) by high-performance liquid chromatography; their identities were subsequently confirmed by UV absorbance, nuclear magnetic resonance, and mass spectra. The conversion of 1 mol of nicotinate to 6-hydroxynicotinate consumed 0.5 mol of O2. From 18O isotopic incorporation experiments, water was the hydroxyl-equivalent source. A nicotinate hydroxylase activity proved to be cell wall-membrane associated; this activity served as direct electron donor (not indirect via NADP+) to O2 via membrane electron transport. These catabolic reactions have not previously been witnessed together in the same organism. A. caulinodans nicotinate catabolism seems coupled to N2 fixation, although the explicit mechanism of this coupling remains to be determined.


Anticonvulsant effect of anterior thalamic high frequency electrical stimulation in the rat.
Evidence suggests that a specific subcortical pathway synaptically linking the anterior thalamic nuclear complex (AN) to the hypothalamus and midbrain is important in the expression of pentylenetetrazol (PTZ) seizures. Perturbation of neuronal activity along this path via focal disruption or chemical inhibition significantly raises seizure threshold. Recent data has demonstrated that focal electrical stimulation within the hypothalamic component of this pathway inhibited seizure expression in a current and frequency dependent fashion. Similar experiments were conducted in the AN to investigate the hypothesis that stimulation of this thalamic nuclear region can prevent the propagation of PTZ seizures between cortical and subcortical regions. Our results indicate that high frequency (100 Hz) stimulation of AN did not alter the expression of low dose PTZ induced cortical bursting but did raise the clonic seizure threshold compared to naive animals or those stimulated at sites near, but not in AN (P < 0.01). Low frequency stimulation (8 Hz) was in contrast, proconvulsant and could induce behavioral arrest responses accompanied by rhythmic high voltage EEG even without PTZ challenge. This data further highlights the role of AN in mediating the expression of seizures and provides experimental support for the concept that this thalamic region may be a promising target for focal stimulation to treat intractable seizures in humans.


Are radiogallium-labelled DOTA-conjugated somatostatin analogues superior to those labelled with other radiometals?
Gallium-68 is a metallic positron emitter with a half-life of 68 min that is ideal for the in vivo use of small molecules, such as [68Ga-DOTA,Tyr3]octreotide, in the diagnostic imaging of somatostatin receptor-positive tumours. In preclinical studies it has shown a striking superiority over its 111In-labelled congener. The purpose of this study was to evaluate whether third-generation somatostatin-based, radiogallium-labelled peptides show the same superiority. Peptides were synthesised on solid phase. The receptor affinity was determined by in vitro receptor autoradiography. The internalisation rate was studied in AR4-2J and hsst-HEK-transfected cell lines. The pharmacokinetics was studied in a rat xenograft tumour model, AR4-2J. All peptides showed high affinities on hsst2, with the highest affinity for the Ga(III)-complexed peptides. On hsst3 the situation was reversed, with a trend towards lower affinity of the Ga(III) peptides. A significantly increased internalisation rate was found in sst2-expressing cells for all 67Ga-labelled peptides. Internalisation into HEK-sst3 was usually faster for the 111In-labelled peptides. No internalisation was found into sst5. Biodistribution studies employing [67Ga-DOTA,1-Nal3]octreotide in comparison to [111In-DOTA,1-Nal3]octreotide and [67Ga-DOTA,Tyr3]octreotide showed a significantly higher and receptor-mediated uptake of the two 67Ga-labelled peptides in the tumour and somatostatin receptor-positive tissues. A patient study illustrated the potential advantage of a broad receptor subtype profile radiopeptide over a high-affinity sst2-selective radiopeptide. This study demonstrates that 67/68Ga-DOTA-octapeptides show distinctly better preclinical, pharmacological performances than the 111In-labelled peptides, especially on sst2-expressing cells and the corresponding animal models. They may be excellent candidates for further development for clinical studies.


The role of indirect photochemical degradation in the environmental fate of pesticides: a review.
Photochemical degradation contributes to the environmental fate of many pesticides in surface waters. A better understanding of the role of direct and indirect photochemical degradation of pesticides is necessary in order to predict their environmental fate and persistence. This review includes all major pesticide classes and focuses on the importance of dissolved organic matter (DOM) as a sensitizer in indirect photodegradation within aquatic systems. Photochemical studies conducted under environmentally relevant conditions (i.e., aqueous solutions with irradiation wavelengths >290 nm) are included. Comparisons are made between observed photodegradation rates in pure or buffered water and in water containing DOM to assess the extent of pesticide susceptibility to DOM-sensitized indirect photolysis. When data is available, the role of specific reactive species in indirect photodegradation is described. While it is possible to assess the relative importance of direct and indirect photodegradation on a pesticide-by-pesticide basis in many cases, it is often difficult to make generalizations based on compound class. Knowledge gaps and inconstancies in the current body of literature are discussed and areas that require additional research are described.


The microcircuits of striatum in silico.
The basal ganglia play an important role in decision making and selection of action primarily based on input from cortex, thalamus, and the dopamine system. Their main input structure, striatum, is central to this process. It consists of two types of projection neurons, together representing 95% of the neurons, and 5% of interneurons, among which are the cholinergic, fast-spiking, and low threshold-spiking subtypes. The membrane properties, soma-dendritic shape, and intrastriatal and extrastriatal synaptic interactions of these neurons are quite well described in the mouse, and therefore they can be simulated in sufficient detail to capture their intrinsic properties, as well as the connectivity. We focus on simulation at the striatal cellular/microcircuit level, in which the molecular/subcellular and systems levels meet. We present a nearly full-scale model of the mouse striatum using available data on synaptic connectivity, cellular morphology, and electrophysiological properties to create a microcircuit mimicking the real network. A striatal volume is populated with reconstructed neuronal morphologies with appropriate cell densities, and then we connect neurons together based on appositions between neurites as possible synapses and constrain them further with available connectivity data. Moreover, we simulate a subset of the striatum involving 10,000 neurons, with input from cortex, thalamus, and the dopamine system, as a proof of principle. Simulation at this biological scale should serve as an invaluable tool to understand the mode of operation of this complex structure. This platform will be updated with new data and expanded to simulate the entire striatum.


Computational Glycobiology: Mechanistic Studies of Carbohydrate-Active Enzymes and Implication for Inhibitor Design.
Carbohydrate-active enzymes (CAZymes) are families of essential and structurally related enzymes, which catalyze the creation, modification, and degradation of glycosidic bonds in carbohydrates to maintain essentially all kingdoms of life. CAZymes play a key role in many biological processes underpinning human health and diseases (e.g., cancer, diabetes, Alzheimer's diseases, AIDS) and have thus emerged as important drug targets in the fight against pathogenesis. The realization of the full potential of CAZymes remains a significant challenge, relying on a deeper understanding of the molecular mechanisms of catalysis. Considering numerous unsettled questions in the literature, while with a large amount of structural, kinetic, and mutagenesis data available for CAZymes, there is a pressing need and an abundant opportunity for collaborative computational and experimental investigations with the aim to unlock the secrets of CAZyme catalysis at an atomic level. In this review, we briefly survey key methodology development in computational studies of CAZyme catalysis. This is complemented by selected case studies highlighting mechanistic insights provided by computational glycobiology. Implication for inhibitor design by mimicking the transition state is also illustrated for both glycoside hydrolases and glycosyltransferases. The challenges for such studies will be noted and finally an outlook for future directions will be provided.


The economic benefits of malaria elimination: do they include increases in tourism?
Policy makers have speculated that one of the economic benefits of malaria elimination includes increases in foreign direct investment, particularly tourism. This study examines the empirical relationship between the demand for travel and malaria cases in two countries with large tourism industries around the time in which they carried out malaria-elimination campaigns. In Mauritius, this analysis examines historical, yearly tourist arrivals and malaria cases from 1978-1999, accounting for the background secular trend of increasing international travel. In Dominican Republic, a country embarking upon malaria elimination, it employs a time-series analysis of the monthly, international tourist arrivals from 1998-2010 to determine whether the timing of significant deviations in tourist arrivals coincides with malaria outbreaks. While naïve relationships exist in both cases, the results show that the relationships between tourist arrivals and malaria cases are relatively weak and statistically insignificant once secular confounders are accounted for. This suggests that any economic benefits from tourism that may be derived from actively pursuing elimination in countries that have high tourism potential are likely to be small when measured at a national level. Rather, tourism benefits are likely to be experienced with greater impact in more concentrated tourist areas within countries, and future studies should seek to assess these relationships at a regional or local level.


Economic aspects of infertility care: a challenge for researchers and clinicians.
Infertility care has improved remarkably over the last few decades and has received growing attention from health care providers. Several treatments, including expensive options such as Assisted Reproductive Techniques, are now widely available for routine clinical use. In most cases, adoption of these treatments has occurred without robust cost-effective analyses. IVF for unexplained infertility and ICSI in the absence of semen abnormalities are two examples of this gradual technology creep. More in-depth economic analyses in the field of infertility are undoubtedly warranted. However, performing these analyses is challenging because infertility care poses a number of unique challenges. Studies of cost-effectiveness are open to criticism because there is a lack of consensus about the outcomes of choice and the appropriate perspective. The use of quality adjusted life years (QALYs) to allow comparisons with other clinical conditions is also controversial because the value associated with infertility care cannot be easily captured in QALYs. Moreover, their use triggers the crucial question of whose QALYs merit consideration-an individual's, a couple's or a child's. In conclusion, economic analysis in infertility represents a peculiar but crucial challenge. If management of infertility is to become an integral part of publicly or privately funded health care systems worldwide, better quality data and a shared vision about the costs and benefits of infertility treatments are needed.


Spatial parcellations, spectral filtering, and connectivity measures in fMRI: Optimizing for discrimination.
The analysis of Functional Connectivity (FC) is a key technique of fMRI, having been used to distinguish brain states and conditions. While many approaches to calculating FC are available, there have been few assessments of their differences, making it difficult to choose approaches, and compare results. Here, we assess the impact of methodological choices on discriminability, using a fully controlled data set of continuous active states involving basic visual and motor tasks, providing robust localized FC changes. We tested a range of anatomical and functional parcellations, including the AAL atlas, parcellations derived from the Human Connectome Project and Independent Component Analysis (ICA) of many dimensionalities. We measure amplitude, covariance, correlation, and regularized partial correlation under different temporal filtering choices. We evaluate features derived from these methods for discriminating states using MVPA. We find that multidimensional parcellations derived from functional data performed similarly, outperforming an anatomical atlas, with correlation and partial correlation (p < .05, FDR). Partial correlation, with appropriate regularization, outperformed correlation. Amplitude and covariance generally discriminated less well, although gave good results with high-dimensionality ICA. We found that discriminative FC properties are frequency specific; higher frequencies performed surprisingly well under certain configurations of atlas choices and dependency measures, with ICA-based parcellations revealing greater discriminability at high frequencies compared to other parcellations. Methodological choices in FC analyses can have a profound impact on results and can be selected to optimize accuracy, interpretability, and sharing of results. This work contributes to a basis for consistent selection of approaches to estimating and analyzing FC.


Simultaneous measurement of thrombin generation and fibrin formation in whole blood under flow conditions.
Assays based on the formation of thrombin and fibrin are frequently used, and results are considered exchangeable in research/clinical settings. However, thrombin generation and fibrin formation do not always go hand in hand and flow profoundly influences thrombus formation. We describe the technical/clinical evaluation of an assay to simultaneously measure thrombin generation and fibrin formation under conditions of flow. Introduction of a fluorometer into a 'cone and base principle'-based rheometer allowed the measurement of thrombin generation (using a thrombin-sensitive substrate) and fibrin formation (changes in viscosity), while applying a linear shear flow. Increasing shear rates inversely related with thrombin generation and fibrin formation. Increasing fibrinogen concentrations in defibrinated plasma resulted in increased thrombin generation and fibrin formation. In pre-operative samples of 70 patients undergoing cardiothoracic surgery, fibrin formation and thrombin generation parameters correlated with fibrinogen content, rotational thromboelastometry (ROTEM) and whole blood Calibrated Automated Thrombinography (CAT) parameters, respectively. Upon dividing patients into two groups based on the median clot strength, a significant difference in perioperative/total blood loss was established. In conclusion, we clinically evaluated a method capable of simultaneously measuring thrombin generation and fibrin formation in plasma/whole blood under continuous flow, rendering our method one step closer to physiology. Importantly, our test proved to be indicative for the amount of blood loss during/after cardiothoracic surgery.


Mixed multilayered vertical heterostructures utilizing strained monolayer WS2.
Creating alternating layers of 2D materials forms vertical heterostructures with diverse electronic and opto-electronic properties. Monolayer WS2 grown by chemical vapour deposition can have inherent strain due to interactions with the substrate. The strain modifies the band structure and properties of monolayer WS2 and can be exploited in a wide range of applications. We demonstrate a non-aqueous transfer method for creating vertical stacks of mixed 2D layers containing a strained monolayer of WS2, with Boron Nitride and Graphene. The 2D materials are all grown by CVD, enabling large area vertical heterostructures to be formed. WS2 monolayers grown by CVD directly on Si substrates with SiO2 surface are easily washed off by water and this makes aqueous based transfer methods challenging for creating vertical stacks on the growth substrate. 2D hexagonal Boron Nitride films are used to provide an insulating layer that limits interactions with a top graphene layer and preserve the strong photoluminescence from the WS2. This transfer method is suitable for layer by layer control of 2D material vertical stacks and is shown to be possible for all CVD grown samples, which opens up pathways for the rapid large scale fabrication of vertical heterostructure systems with atomic thickness depth control and large area coverage.