Outbreaks of Acute Gastroenteritis Transmitted by Person-to-Person Contact, Environmental Contamination, and Unknown Modes of Transmission--United States, 2009-2013.
Acute gastroenteritis (AGE) is a major cause of illness in the United States, with an estimated 179 million episodes annually. AGE outbreaks propagated through direct person-to-person contact, contaminated environmental surfaces, and unknown modes of transmission were not systematically captured at the national level before 2009 and thus were not well characterized. 2009-2013. The National Outbreak Reporting System (NORS) is a voluntary national reporting system that supports reporting of all waterborne and foodborne disease outbreaks and all AGE outbreaks resulting from transmission by contact with contaminated environmental sources, infected persons or animals, or unknown modes. Local, state, and territorial public health agencies within the 50 U.S. states, the District of Columbia (DC), five U.S. territories, and three Freely Associated States report outbreaks to CDC via NORS using a standard online data entry system. A total of 10,756 AGE outbreaks occurred during 2009-2013, for which the primary mode of transmission occurred through person-to-person contact, environmental contamination, and unknown modes of transmission. NORS received reports from public health agencies in 50 U.S. states, DC, and Puerto Rico. These outbreaks resulted in 356,532 reported illnesses, 5,394 hospitalizations, and 459 deaths. The median outbreak reporting rate for all sites in a given year increased from 2.7 outbreaks per million population in 2009 to 11.8 outbreaks in 2013. The etiology was unknown in 31% (N = 3,326) of outbreaks. Of the 7,430 outbreaks with a suspected or confirmed etiology reported, norovirus was the most common, reported in 6,223 (84%) of these outbreaks. Other reported suspected or confirmed etiologies included Shigella (n = 332) and Salmonella (n = 320). Outbreaks were more frequent during the winter, with 5,716 (53%) outbreaks occurring during December-February, and 70% of the 7,001 outbreaks with a reported setting of exposure occurred in long-term-care facilities (n = 4,894). In contrast, 59% (n = 143) of shigellosis outbreaks, 36% (n = 30) of salmonellosis outbreaks, and 32% (n = 84) of other or multiple etiology outbreaks were identified in child care facilities. NORS is the first U.S. surveillance system that provides national data on AGE outbreaks spread through person-to-person contact, environmental contamination, and unknown modes of transmission. The increase in reporting rates during 2009-2013 indicates that reporting to NORS improved notably in the 5 years since its inception. Norovirus is the most commonly reported cause of these outbreaks and, on the basis of epidemiologic data, might account for a substantial proportion of outbreaks without a reported etiology. During 2009-2013, norovirus accounted for most deaths and health care visits in AGE outbreaks spread through person-to-person contact, environmental contamination, and unknown modes of transmission. Recommendations for prevention and control of AGE outbreaks transmitted through person-to-person contact, environmental contamination, and unknown modes of transmission depend primarily on appropriate hand hygiene, environmental disinfection, and isolation of ill persons. NORS surveillance data can help identify priority targets for the development of future control strategies, including hygiene interventions and vaccines, and help monitor the frequency and severity of AGE outbreaks in the United States. Ongoing study of these AGE outbreaks can provide a better understanding of certain pathogens and their modes of transmission. For example, certain reported outbreak etiologies (e.g., Salmonella) are considered primarily foodborne pathogens but can be transmitted through multiple routes. Similarly, further examination of outbreaks of unknown etiology could help identify barriers to making an etiologic determination, to analyze clinical and epidemiologic clues suggestive of a probable etiology, and to discover new and emerging etiologic agents. Outbreak reporting to NORS has improved substantially since its inception, and further outreach efforts and system improvements might facilitate additional increases in the number and completeness of reports to NORS.


Effects of thermal modulation on diffraction in liquid crystal composite gratings.
A microperiodic structure composed of polymer and liquid crystal (LC) phases, called holographic polymer dispersed LC, is fabricated by a photo-induced phase separation technique using LC composites with different physical properties, such as refractive indices and clearing points. Effects of thermal modulation on diffraction properties of LC composite gratings are experimentally investigated in the viewpoints of polarization and temperature dependences. The diffractions based on the change of refractive index induced by the nematic-isotropic transition of LCs with the increase of temperature are applied for a holographic image reconstruction.


A comparison of open and laparoscopic cholecystectomy done by a surgeon in training.
Laparoscopic cholecystectomy has virtually replaced conventional open cholecystectomy as the gold standard for symptomatic cholelithiasis. The laparoscopic approach brings numerous advantages at the expense of higher complication rates, especially in training facilities. This comparative 18-month review examines the outcomes of 52 cholecystectomies performed by a single surgical resident at the University Hospital of the West Indies--a teaching hospital in Jamaica. The advantages of laparoscopic cholecystectomy have been demonstrated and it has been found to be safe and effective in this training facility.


Serum endocan levels in patients with stable COPD.
Endothelial cell specific molecule-1, also called as endocan, is a dermatan sulfate proteoglycan, which is expressed by endothelial cells in alveolar walls of the lung and kidney. High endocan levels are found associated with endothelial dysfunction and inflammation. We hypothesize that endocan level is also high in COPD due to systemic inflammation and endothelial dysfunction. We aimed to investigate the expression of endocan in patients with stable COPD. The study included patients with COPD and control subjects. COPD patients were classified according to the Global Initiative for Chronic Obstructive Lung Disease (GOLD) 2017 criteria. Demographics, body mass index, smoking history, and comorbidities were recorded. Endocan levels of COPD patients and controls were compared. Totally, 88 subjects (47 stable COPD patients, 41 controls) were evaluated. Endocan levels were significantly higher in COPD patients than control group (860.1±259.8 vs 647.3±316.9 pg/mL, P=0.001). There was no relationship between GOLD COPD categories and endocan levels. Also endocan levels were similar between COPD patients with or without hypoxemia. Serum endocan level was significantly higher in patients with stable COPD. Further studies should be performed to better understand the relationship between endocan and COPD.


Identification and characterization of the anti-methicillin-resistant Staphylococcus aureus WAP-8294A2 biosynthetic gene cluster from Lysobacter enzymogenes OH11.
Lysobactor enzymogenes strain OH11 is an emerging biological control agent of fungal and bacterial diseases. We recently completed its genome sequence and found it contains a large number of gene clusters putatively responsible for the biosynthesis of nonribosomal peptides and polyketides, including the previously identified antifungal dihydromaltophilin (HSAF). One of the gene clusters contains two huge open reading frames, together encoding 12 modules of nonribosomal peptide synthetases (NRPS). Gene disruption of one of the NRPS led to the disappearance of a metabolite produced in the wild type and the elimination of its antibacterial activity. The metabolite and antibacterial activity were also affected by the disruption of some of the flanking genes. We subsequently isolated this metabolite and subjected it to spectroscopic analysis. The mass spectrometry and nuclear magnetic resonance data showed that its chemical structure is identical to WAP-8294A2, a cyclic lipodepsipeptide with potent anti-methicillin-resistant Staphylococcus aureus (MRSA) activity and currently in phase I/II clinical trials. The WAP-8294A2 biosynthetic genes had not been described previously. So far, the Gram-positive Streptomyces have been the primary source of anti-infectives. Lysobacter are Gram-negative soil/water bacteria that are genetically amendable and have not been well exploited. The WAP-8294A2 synthetase represents one of the largest NRPS complexes, consisting of 45 functional domains. The identification of these genes sets the foundation for the study of the WAP-8294A2 biosynthetic mechanism and opens the door for producing new anti-MRSA antibiotics through biosynthetic engineering in this new source of Lysobacter.


Restorative aspects of oral cancer reconstruction.
The oral rehabilitation of head and neck cancer patients is a vital and pivotal part of their care pathway, helping to restore function, aesthetics and improve patient well-being and quality of life. Patient-reported outcome data demonstrates that such patients consider that many of their continued and unmet needs relate to their oral and dental rehabilitation. It is therefore incumbent on all dental team members to be aware of the entire spectrum of restorative management options that can be utilised in both the prevention and rehabilitation of this patient cohort. The literature highlights that maxillary and mid-face tumours are challenging, as patients often suffer both profound functional and aesthetic consequences with significant physiological and psychological impact. Conversely, mandibular and floor of mouth tumours often result in issues primarily affecting oral function. This paper will summarise commonly encountered functional and aesthetic issues facing the restorative dental team and highlight widespread preventive challenges.


Improving Collaborative Behaviour Planning in Adult Auditory Rehabilitation: Development of the I-PLAN Intervention Using the Behaviour Change Wheel.
The consequences of poorly managed hearing loss can be ameliorated with hearing aid use but rates of use are sub-optimal. The impact of audiologist behaviour on subsequent use, particularly over the long term, is unknown. This study aimed to describe the role of the behaviour change wheel in developing an intervention to introduce and embed particular clinical behaviours into adult hearing aid fitting consultations, within the framework of the Medical Research Council guidance on complex interventions. Following the steps of the behaviour change wheel, audiologist behaviours that might influence hearing aid use were identified based on a systematic review and qualitative work with audiologists. An analysis, using the COM-B model, identified potential drivers of the target behaviours. This was used to select intervention functions and behaviour change techniques likely to influence behaviour in this context. The target behaviours were as follows: giving information about the benefits of hearing aid use and the negative consequences of non-use, providing prompts for use and engaging in collaborative behavioural planning for use. The behavioural analysis suggested that psychological capability, opportunity and motivation were potential drivers of these behaviours. The intervention functions of education, coercion, training, environmental restructuring, modelling and enablement were selected and combined to develop a single complex intervention that seeks to address the target behaviours.


Veterans' Perspectives on Fitbit Use in Treatment for Post-Traumatic Stress Disorder: An Interview Study.
The increase in availability of patient data through consumer health wearable devices and mobile phone sensors provides opportunities for mental health treatment beyond traditional self-report measurements. Previous studies have suggested that wearables can be effectively used to benefit the physical health of people with mental health issues, but little research has explored the integration of wearable devices into mental health care. As such, early research is still necessary to address factors that might impact integration including patients' motivations to use wearables and their subsequent data. The aim of this study was to gain an understanding of patients' motivations to use or not to use wearables devices during an intensive treatment program for post-traumatic stress disorder (PTSD). During this treatment, they received a complementary Fitbit. We investigated the following research questions: How did the veterans in the intensive treatment program use their Fitbit? What are contributing motivators for the use and nonuse of the Fitbit? We conducted semistructured interviews with 13 veterans who completed an intensive treatment program for PTSD. We transcribed and analyzed interviews using thematic analysis. We identified three major motivations for veterans to use the Fitbit during their time in the program: increase self-awareness, support social interactions, and give back to other veterans. We also identified three major reasons certain features of the Fitbit were not used: lack of clarity around the purpose of the Fitbit, lack of meaning in the Fitbit data, and challenges in the veteran-provider relationship. To integrate wearable data into mental health treatment programs, it is important to understand the patient's perspectives and motivations in using wearables. We also discuss how the military culture and PTSD may have contributed to our participants' behaviors and attitudes toward Fitbit usage. We conclude with possible approaches for integrating patient-generated data into mental health treatment settings that may address the challenges we identified.


Oral Health Symptoms and Cognitive Function Among US Community-Dwelling Chinese Older Adults.
Limited research is available on the relationship between oral health symptoms and cognitive function among community-dwelling US Chinese older adults. The purpose of this study was to examine the associations between tooth/gum symptoms and changes in cognitive function. Two-wave epidemiological study. Population Study of Chinese Elderly in Chicago (PINE). US Chinese older adults (N = 2713; mean age = 72.6 y; 58.4% women). We selected self-reported oral (tooth and gum) symptoms as independent variables. To examine changes in cognitive function (wave 2: Baseline), we chose the following three domains: episodic memory (East Boston Memory Test); executive function (Symbol Digit Modalities Test); and working memory (Digit Span Backwards). In addition, we assessed global cognitive function by constructing a composite measure. At baseline, 1297 participants (47.8%) reported having teeth symptoms, and 513 participants (18.9%) reported having gum symptoms. Adjusting for sociodemographic and health-related characteristics, participants who reported having teeth symptoms at baseline experienced their global cognition decrease by 0.07 units (estimate = -0.07; p = .003) and their episodic memory decrease by 0.07 units (estimate = -0.07; p = .026). Participants who reported having teeth symptoms at baseline experienced a faster rate of decline in global cognition for every additional year (estimate = 0.02; p = .047). However, this effect disappeared once we adjusted for all covariates (estimate = 0.02; p = .069). We found no significant relationship between baseline gum symptoms and change of cognitive function. Having teeth symptoms was associated with a decline in cognitive function among US Chinese older adults. Developing policy measures aimed at ameliorating health and improving cognition in this high-risk fast-growing population in the United States would need to include oral health preventive and dental care services. J Am Geriatr Soc 67:S532-S537, 2019.


Fertility control? Middle-aged Australian women's retrospective reports of their pregnancies.
To assess middle-aged Australian women's retrospective reports of how intended and wanted were their pregnancies, and the degree of happiness associated with these pregnancies. A self-administered questionnaire was sent to 1000 participants in the Mid-Age cohort of the Australian Longitudinal Study on Women's Health. Responses from 811 women (81%) showed that, although 32% of first pregnancies were unplanned and 29% were unwanted, most women recall being happy with their pregnancies and termination rates were very low. The second pregnancy was the most planned and wanted and associated with the highest levels of happiness. While the majority of middle-aged women report having been happy to be pregnant, and the majority of pregnancies are described retrospectively as planned and wanted, a significant proportion of pregnancies are unwanted, unplanned or resulting from unintended contraceptive failure. The data support the continuing need for widely available, affordable and sensitive fertility control services.


Different contributions of the angiotensin-converting enzyme C-domain and N-domain in subjects with the angiotensin-converting enzyme II and DD genotype.
Angiotensin-converting enzyme (ACE) insertion/deletion (I/D) polymorphism-related differences in ACE concentration do not result in differences in angiotensin levels. To investigate whether this relates to differences in the contribution of the ACE C-domain and N-domain, we quantified, using the C-domain-selective inhibitors quinaprilat and RXPA380, and the N-domain-selective inhibitor RXP407, the contribution of both domains to the metabolism of angiotensin I, bradykinin, the C-domain-selective substrate Mca-BK(1-8), and the N-domain-selective substrate Mca-Ala in serum of IIs, DDs, and 'hyperACE' subjects (i.e., subjects with increased ACE due to enhanced shedding). During incubation with angiotensin I, the highest angiotensin II levels were observed in sera with the highest ACE activity. This confirms that ACE is rate-limiting with regard to angiotensin II generation. C-domain-selective concentrations of quinaprilat fully blocked angiotensin I-II conversion in DDs, whereas additional N-domain blockade was required to fully block conversion in IIs. Both domains contributed to bradykinin hydrolysis in all subjects, and the inhibition profile of RXP407 when using Mca-Ala was identical in IIs and DDs. In contrast, the RXPA380 concentrations required to block C-domain activity when using Mca-BK (1-8) were three-fold higher in IIs than DDs. The contributions of the C-domain and N-domain differ between DDs and IIs, and RXPA380 is the first inhibitor capable of distinguishing D-allele ACE from I-allele ACE. The lack of angiotensin II accumulation in DDs in vivo is not because of the often quoted concept that ACE is a nonrate-limiting enzyme. It may relate to the fact that in IIs both the N-domain and C- domain generate angiotensin II, whereas in DDs only the C-domain converts angiotensin I.


Deep mutational scanning of hemagglutinin helps predict evolutionary fates of human H3N2 influenza variants.
Human influenza virus rapidly accumulates mutations in its major surface protein hemagglutinin (HA). The evolutionary success of influenza virus lineages depends on how these mutations affect HA's functionality and antigenicity. Here we experimentally measure the effects on viral growth in cell culture of all single amino acid mutations to the HA from a recent human H3N2 influenza virus strain. We show that mutations that are measured to be more favorable for viral growth are enriched in evolutionarily successful H3N2 viral lineages relative to mutations that are measured to be less favorable for viral growth. Therefore, despite the well-known caveats about cell-culture measurements of viral fitness, such measurements can still be informative for understanding evolution in nature. We also compare our measurements for H3 HA to similar data previously generated for a distantly related H1 HA and find substantial differences in which amino acids are preferred at many sites. For instance, the H3 HA has less disparity in mutational tolerance between the head and stalk domains than the H1 HA. Overall, our work suggests that experimental measurements of mutational effects can be leveraged to help understand the evolutionary fates of viral lineages in nature-but only when the measurements are made on a viral strain similar to the ones being studied in nature.


The Fragility of Statistically Significant Findings From Randomized Trials in Sports Surgery: A Systematic Survey.
High-quality, evidence-based orthopaedic care relies on the generation and translation of robust research evidence. The Fragility Index is a novel method for evaluating the robustness of statistically significant findings from randomized controlled trials (RCTs). It is defined as the minimum number of patients in 1 arm of a trial that would have to change status from a nonevent to an event to alter the results of the trial from statistically significant to nonsignificant. To calculate the Fragility Index of statistically significant results from clinical trials in sports medicine and arthroscopic surgery to characterize the robustness of the RCTs in these fields. A search was conducted in Medline, EMBASE, and PubMed for RCTs related to sports medicine and arthroscopic surgery from January 1, 2005, to October 30, 2015. Two reviewers independently assessed titles and abstracts for study eligibility, performed data extraction, and assessed risk of bias. The Fragility Index was calculated using the Fisher exact test for all statistically significant dichotomous outcomes from parallel-group RCTs. Bivariate correlation was performed to evaluate associations between the Fragility Index and trial characteristics. A total of 48 RCTs were included. The median sample size was 64 (interquartile range [IQR], 48.5-89.5), and the median total number of outcome events was 19 (IQR, 10-27). The median Fragility Index was 2 (IQR, 1-2.8), meaning that changing 2 patients from a nonevent to an event in the treatment arm changed the result to a statistically nonsignificant result, or P ≥ .05. Most statistically significant RCTs in sports medicine and arthroscopic surgery are not robust because their statistical significance can be reversed by changing the outcome status on only a few patients in 1 treatment group. Future work is required to determine whether routine reporting of the Fragility Index enhances clinicians' ability to detect trial results that should be viewed cautiously.


Impact of 1% malic acid spray on the oral health-related quality of life of patients with xerostomia.
Dry mouth sensation, also known as xerostomia, is a common clinical problem with an increasing prevalence. Although recent studies have reported promissory results of malic acid, none have evaluated the impact of malic acid on the oral health-related quality of life (OHRQoL) of patients with xerostomia. Thus, this study aimed to evaluate the impact of 1% malic acid, combined with fluoride and xylitol, on the OHRQoL of patients with xerostomia. We enrolled 70 patients and randomly allocated them into two groups: the intervention group (applied topical sialogogue with 1% malic acid) and the control group (applied a placebo). We assessed the OHRQoL and severity of xerostomia before and after treatment with the Spanish version of the Oral Health Impact Profile-14 questionnaire (OHIP-14sp) and a visual analogue scale (VAS), respectively. In addition, stimulated and non-stimulated salivary flow rates before and after treatments were also measured. In total, 60 patients completed the study. According to the VAS, both sprays significantly improved dry mouth sensation (P < 0.001). However, OHIP-14sp total scores decreased significantly in the intervention group from 20.8 ± 10.4 to 16.5 ± 9.5 (P < 0.001), indicating an improvement in the OHRQoL. No significant differences were observed in the control group (P > 0.05). Furthermore, non-stimulated salivary flow rates significantly increased in the intervention group from 0.25 ± 0.22 to 0.33 ± 0.33 mL/min (P < 0.001). Overall, this study demonstrated that malic acid improves the OHRQoL and dry mouth sensation in patients with xerostomia.


Shortening of 3' UTRs in most cell types composing tumor tissues implicates alternative polyadenylation in protein metabolism.
During pre-mRNA maturation 3' end processing can occur at different polyadenylation sites in the 3' untranslated region (3' UTR) to give rise to transcript isoforms that differ in the length of their 3' UTRs. Longer 3' UTRs contain additional cis-regulatory elements that impact the fate of the transcript and/or of the resulting protein. Extensive alternative polyadenylation (APA) has been observed in cancers, but the mechanisms and roles remain elusive. In particular, it is unclear whether the APA occurs in the malignant cells or in other cell types that infiltrate the tumor. To resolve this, we developed a computational method, called SCUREL, that quantifies changes in 3' UTR length between groups of cells, including cells of the same type originating from tumor and control tissue. We used this method to study APA in human lung adenocarcinoma (LUAD). SCUREL relies solely on annotated 3' UTRs and on control systems such as T cell activation, and spermatogenesis gives qualitatively similar results at much greater sensitivity compared to the previously published scAPA method. In the LUAD samples, we find a general trend toward 3' UTR shortening not only in cancer cells compared to the cell type of origin, but also when comparing other cell types from the tumor vs. the control tissue environment. However, we also find high variability in the individual targets between patients. The findings help in understanding the extent and impact of APA in LUAD, which may support improvements in diagnosis and treatment.


Perceived exertion and performance modulation: effects of caffeine ingestion and subject expectation.
It is well established that caffeine has ergogenic effects on endurance performance. This evidence often comes from studies in which subjects receive either caffeine or placebo in double-blind, randomized and counterbalanced order. Here, we propose a new methodology which aims to estimate the effects of participant expectancy of ergogenic or anti-ergogenic effects on performance. Sixteen physically active participants (non-athletes engaged in systematic physical training >3 months, at least three times a week) performed three 30-minute running tests after being told they would be provided with either a harmful treatment (lactic acid), a beneficial treatment (caffeine) or a placebo. In each blinded case, subjects were given caffeine. The velocity and Rating of Perceived Exertion (RPE) during the time trial were examined in light of the participant's expectancy before and after the endurance event using Bayesian multilevel models. For pre-exercise expectancy, there is a 92% probability that caffeine expectation decreases RPE (posterior median±SD -0.65±0.36) and a 79% probability that lactic acid expectation increases RPE (posterior median±SD 0.58±0.47) with expectations for placebo and 'not sure' falling in between (posterior median±SD: -0.37±0.32 and -0.22±0.37, respectively). In general, our interventions suggest an 81% probability that caffeine lowers RPE. However, there was no effect of caffeine supplementation on running velocity (median±SD 0.04±0.08 km.h-1). When a participant believed they are under a potentially positive treatment, their RPE decreased but if they believed themselves to be under a harmful treatment, their RPE increased, regardless of the actual positive intervention; neither caffeine nor the expectancy of a particular intervention improved actual performance as measured by running velocity in a 30-minute period.


A comparison of health access between permanent residents, undocumented immigrants and refugee claimants in Toronto, Canada.
Understanding the immigrant experience accessing healthcare is essential to improving their health. This qualitative study reports on experiences seeking healthcare for three groups of immigrants in Toronto, Canada: permanent residents, refugee claimants and undocumented immigrants. Undocumented immigrants who are on the Canadian Border Services Agency deportation list are understudied in Canada due to their precarious status. This study will examine the vulnerabilities of this particular subcategory of immigrant and contrast their experiences seeking healthcare with refugee claimants and permanent residents. Twenty-one semi-structured, one-on-one qualitative interviews were conducted with immigrants to identify barriers and facilitators to accessing healthcare. The open structure of the interviews enabled the participants to share their experiences seeking healthcare and other factors that were an integral part of their health. This study utilized a community-based participatory research framework. The study identifies seven sections of results. Among them, immigration status was the single most important factor affecting both an individual's ability to seek out healthcare and her experiences when trying to access healthcare. The healthcare seeking behaviour of undocumented immigrants was radically distinct from refugee claimants or immigrants with permanent resident status, with undocumented immigrants being at a greater disadvantage than permanent residents and refugee claimants. Language barriers are also noted as an impediment to healthcare access. An individual's immigration status further complicates their ability to establish relationships with family doctors, access prescriptions and medications and seek out emergency room care. Fear of authorities and the complications caused by the above factors can lead to the most disadvantaged to seek out informal or black market sources of healthcare. This study reaffirmed previous findings that fear of deportation forestalls undocumented immigrants from seeking out healthcare through standard means. The findings bring to light issues not discussed in great depth in the current literature on immigrant health access, the foremost being the immigration status of an individual is a major factor affecting that person's ability to seek, and experience of, healthcare services. Further, that undocumented immigrants have difficulty gaining access to pharmaceuticals and so may employ unregulated means to obtain medication, often with the assistance of a doctor. Also, there exists two streams of healthcare access for undocumented immigrants--from conventional healthcare facilities but also from informal systems delivered mainly through community-based organizations. Finally, within the umbrella term 'immigrant' there appears to be drastically different healthcare utilization patterns and attitudes toward seeking out healthcare between the three subgroups of immigrants addressed by this study.


Supplemental written information improves prenatal counseling: a randomized trial.
To determine if maternal knowledge of prematurity is improved when verbal gestational age-specific counseling is supplemented with written gestational age-specific information. Prospective, randomized study of 60 pregnant participants assessed to be at risk for premature delivery between 23 and 34 weeks' gestation. Counseling in the control group consisted of gestational age-specific verbal information, and counseling in the intervention group consisted of written gestational age-specific information 1 hour before the verbal gestational age-specific information. Both groups completed a Prematurity Knowledge Questionnaire after counseling and the State-Trait Anxiety Inventory before and after counseling. The Prematurity Knowledge Questionnaire consisted of questions regarding short-term problems (immature lungs, intraventricular hemorrhage, retinopathy, feeding problems, infection, apnea, and jaundice), long-term problems (chronic lung disease, postdischarge respiratory infections, visual impairment, hearing impairment, brain damage, and learning and behavior problems), and numerical outcome data (probabilities of survival, survival without significant morbidity, severe intraventricular hemorrhage, severe retinopathy, and chronic lung disease). Knowledge of short-term problems was not statistically different between the intervention (82%) and control groups (67%). Knowledge of long-term problems was better in the intervention (71%) than the control group (45%). Knowledge of numerical data was better in the intervention (48%) than the control group (29%). State-Trait Anxiety Inventory scores decreased after counseling in the intervention group. Supplementation of face-to-face verbal counseling with written information improved knowledge of long-term problems and knowledge of numerical outcome data, and it also decreased anxiety in women expecting a premature delivery.


Pattern Analysis of Sorghum Genotype × Environment Interaction for Leaf, Panicle, and Grain Anthracnose in Mali.
Resistance to anthracnose, caused by Colletotrichum graminicola, in sorghum was identified through field screening at two locations (Samanko and Longorola) in Mali. The occurrence and progress of anthracnose were monitored on 19 sorghum lines plus resistant and susceptible checks in the 1996 to 1998 rainy seasons. Foliar anthracnose severity was assessed at regular intervals throughout the season. Area under the disease progress curve (AUDPC) was calculated for each genotype. Anthracnose severity was also evaluated on the peduncle, rachis and glumes, panicle, and grain. For the characters under study, the site × year and site × year × line interactions accounted for the genotype × environment interactions. Pattern analysis was applied to the environment-standardized matrix of genotype × environment means to analyze these interactions and elucidate genotypic adaptation. None of the lines was completely (hypersensitive) resistant to the disease, but 12 showed high levels of stable resistance to both foliar and panicle anthracnose. Only one was moderately susceptible to both forms of the disease. In addition to identifying varieties that can be grown in zones to which they are adapted, additional genotypes were identified that can serve as sources of resistance in regional breeding programs.


Neural plasticity: how the eye tells the brain about sound location.
Recent studies in owls and ferrets seem to have identified the origin and nature of the visual signals that shape the development of the auditory space map in the midbrain, which ensures that the neural representations of both sensory modalities share the same topographic organization.


Differentiation and stability of T helper 1 and 2 cells derived from naive human neonatal CD4+ T cells, analyzed at the single-cell level.
The development of CD4+ T helper (Th) type 1 and 2 cells is essential for the eradication of pathogens, but can also be responsible for various pathological disorders. Therefore, modulation of Th cell differentiation may have clinical utility in the treatment of human disease. Here, we show that interleukin (IL) 12 and IL-4 directly induce human neonatal CD4- T cells, activated via CD3 and CD28, to differentiate into Th1 and Th2 subsets. In contrast, IL-13, which shares many biological activities with IL-4, failed to induce T cell differentiation, consistent with the observation that human T cells do not express IL-13 receptors. Both the IL-12-induced Th1 subset and the IL-4-induced Th2 subset produce large quantities of IL-10, confirming that human IL-10 is not a typical human Th2 cytokine. Interestingly, IL-4-driven Th2 cell differentiation was completely prevented by an IL-4 mutant protein (IL-4.Y124D), indicating that this molecule acts as a strong IL-4 receptor antagonist. Analysis of single T cells producing interferon gamma or IL-4 revealed that induction of Th1 cell differentiation occurred rapidly and required only 4 d of priming of the neonatal CD4+ T cells in the presence of IL-12. The IL-12-induced Th1 cell phenotype was stable and was not significantly affected when repeatedly stimulated in the presence of recombinant IL-4. In contrast, the differentiation of Th2 cells occurred slowly and required not only 6 d of priming, but also additional restimulation of the primed CD4+ T cells in the presence of IL-4. Moreover, IL-4-induced Th2 cell phenotypes were not stable and could rapidly be reverted into a population predominantly containing Th0 and Th1 cells, after a single restimulation in the presence of IL-12. The observed differences in stability of IL-12- and IL-4-induced human Th1 and Th2 subsets, respectively, may have implications for cytokine-based therapies of chronic disease.


Pharmacological treatment of fibromyalgia.
Fibromyalgia (FM) is a common syndrome characterised by widespread pain and at least 11/18 painful tender points that requires multimodal pharmacological treatment also combined with non-pharmacological therapy. Various drugs currently are available to control the complex and different symptoms reported by patients. Only three drugs (duloxetine, milnacipram, pregabalin) are approved by the American Food and Drug Administration (FDA) and none by the European Medicines Agency (EMEA), consequently, off-label use is habitual in Europe. Most of the drugs improve only one or two symptoms; no drug capable of overall symptom control is yet available. Furthermore, different classes of drugs with different mechanisms of action are used off-label, including tricyclic antidepressants (TCAs), selective serotonin reuptake inhibitors (SSRIs), serotonin norepinephrine reuptake inhibitors (SNRIs), opioids, non-steroidal anti-inflammatory drugs (NSAIDs), growth hormone, corticosteroids and sedative hypnotics. As no single drug fully manages FM symptoms, multicomponent therapy should be used from the beginning. Various pharmacological treatments have been used to treat FM with inconclusive results, and gradually increasing low doses is suggested in order to maximise efficacy. The best treatment should be individualised and combined with patient education and non-pharmacological therapy.


Mineralogy and deformation in some lunar samples.
Observations on the mineralogy and deformation in samples of crystalline rocks, breccias, and fines from Tranquillity Base provide evidence for magmatic and impact processes. Overall homogeneity, igneous textures, and absence of xenoliths in the crystalline rocks indicate derivation from a common titanium-rich magma by internal, anorogenic volcanism rather than by impact. Crystallization conditions allowed strong compositional variation in pyroxenes, olivine, and plagioclase and the growth of a new mineral, the iron analog of pyroxmangite. Subsequently, impact produced breccias containing shock-deformed crystals and glasses of varying compositions.


Deconstructing transcriptional heterogeneity in pluripotent stem cells.
Pluripotent stem cells (PSCs) are capable of dynamic interconversion between distinct substates; however, the regulatory circuits specifying these states and enabling transitions between them are not well understood. Here we set out to characterize transcriptional heterogeneity in mouse PSCs by single-cell expression profiling under different chemical and genetic perturbations. Signalling factors and developmental regulators show highly variable expression, with expression states for some variable genes heritable through multiple cell divisions. Expression variability and population heterogeneity can be influenced by perturbation of signalling pathways and chromatin regulators. Notably, either removal of mature microRNAs or pharmacological blockage of signalling pathways drives PSCs into a low-noise ground state characterized by a reconfigured pluripotency network, enhanced self-renewal and a distinct chromatin state, an effect mediated by opposing microRNA families acting on the Myc/Lin28/let-7 axis. These data provide insight into the nature of transcriptional heterogeneity in PSCs.


Aneurysmal bone cyst of the ethmoid sinus: A case report.
Aneurysmal bone cyst is an expansile bone lesion, non-neoplastic in nature, occurring most commonly in long bones. It is uncommon in facial bones and exceptionally rare in ethmoid bone. Ten cases of aneurysmal bone cysts of ethmoid bone have been reported so far. A young adolescent presented with decreased vision and pain in the right eye. MRI revealed an expansile lesion having conspicuous fluid levels with a multiloculated appearance in the right ethmoid bone extending to the right orbit. CT was done to characterize better bone details. Both biopsy of the lesion and histopathology of resected specimen confirmed aneurysmal bone cyst. A characteristic appearance on MRI and CT examinations helped to confidently diagnose a relatively common lesion in an exceedingly rare location.


Efficacy of the nicotine inhaler in smoking reduction: A double-blind, randomized trial.
Many smokers are not ready to quit but are interested in changing their smoking behavior, particularly if such a change is associated with a reduction in health risk. The present study evaluated the efficacy of the nicotine inhaler in reducing smoking. Exploratory studies assessed whether reduction in smoking was associated with reduction in markers of disease risk. A total of 429 healthy smokers (smoking at least 20 cigarettes/day) were randomly assigned to either nicotine-containing or placebo inhalers, which subjects were allowed to use ad libitum for up to 1 year. The nicotine inhaler was significantly superior to placebo in achieving reduction in daily cigarette consumption by at least 50% after 4 months, compared with baseline (18% vs. 8%, p = .004). Active treatment promoted smoking cessation: 8% of subjects in the nicotine group and 1% in the placebo group were abstinent at month 15. Throughout the study, smoking reduction, per se, independent of treatment group, was associated with a statistically significant decrease in exhaled carbon monoxide and serum cotinine and thiocyanate. Smoking reduction also improved established risk markers for cardiovascular disease over 4 months. The incidence of adverse events did not differ significantly between the active and placebo groups. The most common treatment-related adverse events were throat irritation and cough. In conclusion, the nicotine inhaler can help smokers who are unable or unwilling to quit to reduce daily cigarette consumption, which may be a health benefit on its own and may further promote quitting.


The central role of comorbidity in predicting ambulatory care sensitive hospitalizations.
Ambulatory care sensitive hospitalizations (ACSHs) are commonly used as measures of access to and quality of care. They are defined as hospitalizations for certain acute and chronic conditions; yet, they are most commonly used in analyses comparing different groups without adjustment for individual-level comorbidity. We present an exploration of their roles in predicting ACSHs for acute and chronic conditions. Using 1998-99 US Medicare claims for 1 06 930 SEER-Medicare control subjects and 1999 Area Resource File data, we modelled occurrence of acute and chronic ACSHs with logistic regression, examining effects of different predictors on model discriminatory power. Flags for the presence of a few comorbid conditions-congestive heart failure, chronic obstructive pulmonary disease, diabetes, hypertension and, for acute ACSHs, dementia-contributed virtually all of the discriminative ability for predicting ACSHs. C-statistics were up to 0.96 for models predicting chronic ACSHs and up to 0.87 for predicting acute ACSHs. C-statistics for models lacking comorbidity flags were lower, at best 0.73, for both acute and chronic ACSHs. Comorbidity is far more important in predicting ACSH risk than any other factor, both for acute and chronic ACSHs. Imputations about quality and access should not be made from analyses that do not control for presence of important comorbid conditions. Acute and chronic ACSHs differ enough that they should be modelled separately. Unaggregated models restricted to persons with the relevant diagnoses are most appropriate for chronic ACSHs.


Accuracy of pedicle screw placement for upper and middle thoracic pathologies without coronal plane spinal deformity using conventional methods.
There is a growing interest in the implantation of pedicle screws into the middle and upper thoracic spine. However, usage of the technique is still under debate for these levels because the pedicles in these regions are small and exhibit a high degree of inter- and intraspecimen variability. Twenty-four consecutive patients treated for upper and middle thoracic pathologies by pedicle screw instrumentation under biplanar or uniplanar fluoroscopy were evaluated retrospectively. The rate of screw misplacement on postoperative computerized tomography and complications caused by misplaced screws were determined. In 24 cases, a total of 113 upper-middle thoracic pedicle screws were inserted. Fifty-one of them were inserted between T2 and T5 with guidance of biplanar fluoroscopy, and 62 were inserted between T6 and T8 with uniplanar fluoroscopy. The rate of misplacement was 20.3% for 113 screws (27.4% for T2 to T5 screws, and 14.5% for 62 T6 to T8 screws). Four screws were incorrectly inserted which could have clinical significance (3.5%), and 1 of them required revision. The difference between the upper and middle thoracic screws was not statistically significant (P=0.089). Screw insertion with laminectomy did not cause significant difference in both upper and middle thoracic regions. Only 2 complications were seen because of screw insertion. Pedicle screw insertion with guidance of fluoroscopy may be a reliable and safe method for upper and middle thoracic pathologies.


Pituitary Hormones and Orofacial Pain.
Clinical and basic research on regulation of pituitary hormones, extra-pituitary release of these hormones, distribution of their receptors and cell signaling pathways recruited upon receptor binding suggests that pituitary hormones can regulate mechanisms of nociceptive transmission in multiple orofacial pain conditions. Moreover, many pituitary hormones either regulate glands that produce gonadal hormones (GnH) or are regulated by GnH. This implies that pituitary hormones may be involved in sex-dependent mechanisms of orofacial pain and could help explain why certain orofacial pain conditions are more prevalent in women than men. Overall, regulation of nociception by pituitary hormones is a relatively new and emerging area of pain research. The aims of this review article are to: (1) present an overview of clinical conditions leading to orofacial pain that are associated with alterations of serum pituitary hormone levels; (2) discuss proposed mechanisms of how pituitary hormones could regulate nociceptive transmission; and (3) outline how pituitary hormones could regulate nociception in a sex-specific fashion. Pituitary hormones are routinely used for hormonal replacement therapy, while both receptor antagonists and agonists are used to manage certain pathological conditions related to hormonal imbalance. Administration of these hormones may also have a place in the treatment of pain, including orofacial pain. Hence, understanding the involvement of pituitary hormones in orofacial pain, especially sex-dependent aspects of such pain, is essential to both optimize current therapies as well as provide novel and sex-specific pharmacology for a diversity of associated conditions.


Carboxylated nanodiamond-mediated CRISPR-Cas9 delivery of human retinoschisis mutation into human iPSCs and mouse retina.
Nanodiamonds (NDs) are considered to be relatively safe carbon nanomaterials used for the transmission of DNA, proteins and drugs. The feasibility of utilizing the NDs to deliver CRISPR-Cas9 system for gene editing has not been clearly studied. Therefore, in this study, we aimed to use NDs as the carriers of CRISPR-Cas9 components designed to introduce the mutation in RS1 gene associated with X-linked retinoschisis (XLRS). ND particles with a diameter of 3 nm were functionalized by carboxylation of the surface and covalently conjugated with fluorescent mCherry protein. Two linear DNA constructs were attached to the conjugated mCherry: one encoded Cas9 endonuclease and GFP reporter, another encoded sgRNA and contained insert of HDR template designed to introduce RS1 c.625C>T mutation. Such nanoparticles were successfully delivered and internalized by human iPSCs and mouse retinas, the efficiency of internalization was significantly improved by mixing with BSA. The delivery of ND particles led to introduction of RS1 c.625C>T mutation in both human iPSCs and mouse retinas. Rs1 gene editing in mouse retinas resulted in several pathological features typical for XLRS, such as aberrant photoreceptor structure. To conclude, our ND-based CRISPR-Cas9 delivery system can be utilized as a tool for creating in vitro and in vivo disease models of XLRS. STATEMENT OF SIGNIFICANCE: X-linked retinoschisis (XLRS) is a prevalent hereditary retinal disease, which is caused by mutations in RS1 gene, whose product is important for structural organization of the retina. The recent development of genome editing techniques such as CRISPR-Cas9 significantly improved the prospects for better understanding the pathology and development of treatment for this disease. Firstly, gene editing can allow development of appropriate in vitro and in vivo disease models; secondly, CRISPR-Cas9 can be applied for gene therapy by removing the disease-causative mutation in vivo. The major prerequisite for these approaches is to develop safe and efficient CRISPR-Cas9 delivery system. In this study, we tested specifically modified nanodiamonds for such a delivery system. We were able to introduce Rs1 mutation into the mouse retina and, importantly, observed several XLRS-specific effects.


Highly conserved residues in the bZIP domain of yeast GCN4 are not essential for DNA binding.
Yeast GCN4 and the Jun oncoprotein are transcriptional activators that bind DNA via a bZIP domain consisting of a leucine zipper dimerization element and an adjacent basic region that directly contacts DNA. Two highly conserved alanines (Ala-238 and Ala-239 in GCN4) and an invariant asparagine (Asn-235) in the basic region have been proposed to play important roles in DNA sequence recognition by bZIP proteins. Surprisingly, these conserved residues can be functionally replaced in GCN4 and in a derivative containing the Jun basic region (Jun-GCN4). The ability of an amino acid to functionally substitute for Asn-235 does not correlate with its preference for assuming the N-cap position of an alpha helix. This finding argues against the proposal of the scissors grip model that the invariant asparagine forms an N cap that permits the basic region to bend sharply and wrap around the DNA. In contrast to a prediction of the induced fork model, the pattern of functional substitutions of the conserved alanines together with the results of uracil interference experiments suggests that Ala-238 and Ala-239 do not make base-specific DNA contacts. Finally, the Jun-GCN4 chimeric proteins appear much more active in vivo than expected from their DNA-binding properties in vitro. The mechanistic and evolutionary implications of these results are discussed.


Involvement of CCR9 at multiple stages of adult T lymphopoiesis.
The chemokine CCL25 is constitutively expressed in the thymus, and its receptor CCR9 is expressed on subsets of developing thymocytes. Nevertheless, the function of CCL25/CCR9 in adult thymopoiesis remains unclear. Here, we demonstrate that purified CCR9(-/-) hematopoietic stem cells are deficient in their ability to generate all major thymocyte subsets including double-negative 1 (DN1) cells in competitive transfers. CCR9(-/-) bone marrow contained normal numbers of lineage(-) Sca-1+c-kit+, common lymphoid progenitors, and lymphoid-primed multipotent progenitors (LMPP), and CCR9(-/-) LMPP showed similar T cell potential as their wild-type (WT) counterparts when cultured on OP9-delta-like 1 stromal cells. In contrast, early thymic progenitor and DN2 thymocyte numbers were reduced in the thymus of adult CCR9(-/-) mice. In fetal thymic organ cultures (FTOC), CCR9(-/-) DN1 cells were as efficient as WT DN1 cells in generating double-positive (DP) thymocytes; however, under competitive FTOC, CCR9(-/-) DP cell numbers were reduced significantly. Similarly, following intrathymic injection into sublethally irradiated recipients, CCR9(-/-) DN cells were out-competed by WT DN cells in generating DP thymocytes. Finally, in competitive reaggregation thymic organ cultures, CCR9(-/-) preselection DP thymocytes were disadvantaged significantly in their ability to generate CD4 single-positive (SP) thymocytes, a finding that correlated with a reduced ability to form TCR-MHC-dependent conjugates with thymic epithelial cells. Together, these results highlight a role for CCR9 at several stages of adult thymopoiesis: in hematopoietic progenitor seeding of the thymus, in the DN-DP thymocyte transition, and in the generation of CD4 SP thymocytes.


Effect of angiotensin-converting enzyme inhibitors and angiotensin II receptor blockers on all-cause mortality, cardiovascular deaths, and cardiovascular events in patients with diabetes mellitus: a meta-analysis.
Angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin II receptor blockers (ARBs) may have different effects on cardiovascular (CV) events in patients with diabetes mellitus (DM). To conduct a meta-analysis to separately evaluate the effects of ACEIs and ARBs on all-cause mortality, CV deaths, and major CV events in patients with DM. DATA SOURCES Data sources included MEDLINE (1966-2012), EMBASE (1988-2012), the Cochrane Central Register of Controlled Trials, conference proceedings, and article reference lists. We included randomized clinical trials reporting the effects of ACEI and ARB regimens for DM on all-cause mortality, CV deaths, and major CV events with an observation period of at least 12 months. Studies were excluded if they were crossover trials. Dichotomous outcome data from individual trials were analyzed using the risk ratio (RR) measure and its 95% CI with random-effects models. We estimated the difference between the estimates of the subgroups according to tests for interaction. We performed meta-regression analyses to identify sources of heterogeneity. Primary end points were all-cause mortality and death from CV causes. Secondary end points were the effects of ACEIs and ARBs on major CV events. Twenty-three of 35 identified trials compared ACEIs with placebo or active drugs (32,827 patients) and 13 compared ARBs with no therapy (controls) (23,867 patients). When compared with controls (placebo/active treatment), ACEIs significantly reduced the risk of all-cause mortality by 13% (RR, 0.87; 95% CI, 0.78-0.98), CV deaths by 17% (0.83; 0.70-0.99), and major CV events by 14% (0.86; 0.77-0.95), including myocardial infarction by 21% (0.79; 0.65-0.95) and heart failure by 19% (0.81; 0.71-0.93). Treatment with ARBs did not significantly affect all-cause mortality (RR, 0.94; 95% CI, 0.82-1.08), CV death rate (1.21; 0.81-1.80), and major CV events (0.94; 0.85-1.01) with the exception of heart failure (0.70; 0.59-0.82). Both ACEIs and ARBs were not associated with a decrease in the risk for stroke in patients with DM. Meta-regression analysis showed that the ACEI treatment effect on all-cause mortality and CV death did not vary significantly with the starting baseline blood pressure and proteinuria of the trial participants and the type of ACEI and DM. Angiotensin-converting enzyme inhibitors reduced all-cause mortality, CV mortality, and major CV events in patients with DM, whereas ARBs had no benefits on these outcomes. Thus, ACEIs should be considered as first-line therapy to limit excess mortality and morbidity in this population.


Epidemiology of seafood-associated infections in the United States.
Seafood is part of a healthful diet, but seafood consumption is not risk-free. Seafood is responsible for an important proportion of food-borne illnesses and outbreaks in the United States. Seafood-associated infections are caused by a variety of bacteria, viruses, and parasites; this diverse group of pathogens results in a wide variety of clinical syndromes, each with its own epidemiology. Some seafood commodities are inherently more risky than others, owing to many factors, including the nature of the environment from which they come, their mode of feeding, the season during which they are harvested, and how they are prepared and served. Prevention of seafood-associated infections requires an understanding not only of the etiologic agents and seafood commodities associated with illness but also of the mechanisms of contamination that are amenable to control. Defining these problem areas, which relies on surveillance of seafood-associated infections through outbreak and case reporting, can lead to targeted research and help to guide control efforts. Coordinated efforts are necessary to further reduce the risk of seafood-associated illnesses. Continued surveillance will be important to assess the effectiveness of current and future prevention strategies.


A hermeneutic phenomenological exploration of 'last resort' in the use of restraint.
Restraining patients is a practice that dates back at least three centuries. In recent years, there has been a mandate and advocacy in various countries for organizations to shift towards the minimization of restraint, whereby its use is only as a 'last resort'. There is growing evidence internationally indicating the negative impact of the use of restraint. However, to date there is no research specifically focusing on trying to understand the concept of 'last resort'. Further insights to explore this concept among mental health nurses are therefore warranted. The empirical research comprised a hermeneutic phenomenological study. By recruiting and interviewing thirteen mental health nurses from across Canada who had experiences of restraint use, the research aimed to generate a deeper understanding of the meanings and lived experiences of the concept of 'last resort'. Data were collected through fifteen in-depth interviews. Data analysis was undertaken through a hermeneutic phenomenological framework based on van Manen's approach and Heideggerian philosophy. Five Heideggerian concepts were used to illuminate 'last resort' in restraint use by mental health nurses - temporality, inauthenticity, thrownness, leaping in and leaping ahead, and mood (fear). Key findings highlight the influence of nurses' past experiences, how nursing staff adopt a collective (rather than individual) approach, and the dependency on knowledge and skills of others in using restraint as a 'last resort'. Overall, the lived experience of 'last resort' is comprised of many elements. This study provides insights and an initial understanding, which is hoped to advance the field of restraint minimization.


Externalizing behavior in early childhood and body mass index from age 2 to 12 years: longitudinal analyses of a prospective cohort study.
Some evidence suggests that obesity and behavior problems are related in children, but studies have been conflicting and have rarely included children under age 4. An association between behavior problems in early childhood and risk for obesity could suggest that a common set of factors contribute to both. Our research objectives were to determine the extent to which externalizing behavior in early childhood is related to body mass index (BMI) in early childhood and through age 12, and to evaluate whether these associations differ by sex and race. Data from the NICHD Study of Early Child Care and Youth Development were analyzed. Externalizing behaviors at 24 months were assessed by mothers using the Child Behavior Checklist. BMI was calculated from measured height and weight assessed 7 times between age 2 and 12 years. Linear mixed effects models were used to assess associations between 24 month externalizing behavior and BMI from 2 to 12 years, calculate predicted differences in BMI, and evaluate effect modification. Externalizing behavior at 24 months was associated with a higher BMI at 24 months and through age 12. Results from a linear mixed effects model, controlling for confounding variables and internalizing behavior, predicted a difference in BMI of approximately 3/4 of a unit at 24 months of age comparing children with high levels of externalizing behavior to children with low levels of externalizing behavior. There was some evidence of effect modification by race; among white children, the average BMI difference remained stable through age 12, but it doubled to 1.5 BMI units among children who were black or another race. Our analyses suggest that externalizing behaviors in early childhood are associated with children's weight status early in childhood and throughout the elementary school years, though the magnitude of the effect is modest.


Ceritinib versus chemotherapy in patients with ALK-rearranged non-small-cell lung cancer previously given chemotherapy and crizotinib (ASCEND-5): a randomised, controlled, open-label, phase 3 trial.
Ceritinib is a next-generation anaplastic lymphoma kinase (ALK) inhibitor, which has shown robust anti-tumour efficacy, along with intracranial activity, in patients with ALK-rearranged non-small-cell lung cancer. In phase 1 and 2 studies, ceritinib has been shown to be highly active in both ALK inhibitor-naive and ALK inhibitor-pretreated patients who had progressed after chemotherapy (mostly multiple lines). In this study, we compared the efficacy and safety of ceritinib versus single-agent chemotherapy in patients with advanced ALK-rearranged non-small-cell lung cancer who had previously progressed following crizotinib and platinum-based doublet chemotherapy. In this randomised, controlled, open-label, phase 3 trial, we recruited patients aged at least 18 years with ALK-rearranged stage IIIB or IV non-small-cell lung cancer (with at least one measurable lesion) who had received previous chemotherapy (one or two lines, including a platinum doublet) and crizotinib and had subsequent disease progression, from 99 centres across 20 countries. Other inclusion criteria were a WHO performance status of 0-2, adequate organ function and laboratory test results, a life expectancy of at least 12 weeks, and having recovered from previous anticancer treatment-related toxicities. We randomly allocated patients (1:1; with blocking [block size of four]; stratified by WHO performance status [0 vs 1-2] and presence or absence of brain metastases) to oral ceritinib 750 mg per day fasted (in 21 day treatment cycles) or chemotherapy (intravenous pemetrexed 500 mg/m2 or docetaxel 75 mg/m2 [investigator choice], every 21 days). Patients who discontinued chemotherapy because of progressive disease could cross over to the ceritinib group. The primary endpoint was progression-free survival, assessed by a masked independent review committee using Response Evaluation Criteria in Solid Tumors 1.1 in the intention-to-treat population, assessed every 6 weeks until month 18 and every 9 weeks thereafter. This trial is registered with ClinicalTrials.gov, number NCT01828112, and is ongoing but no longer recruiting patients. Between June 28, 2013, and Nov 2, 2015, we randomly allocated 231 patients; 115 (50%) to ceritinib and 116 (50%) to chemotherapy (40 [34%] to pemetrexed, 73 [63%] to docetaxel, and three [3%] discontinued before receiving treatment). Median follow-up was 16·5 months (IQR 11·5-21·4). Ceritinib showed a significant improvement in median progression-free survival compared with chemotherapy (5·4 months [95% CI 4·1-6·9] for ceritinib vs 1·6 months [1·4-2·8] for chemotherapy; hazard ratio 0·49 [0·36-0·67]; p<0·0001). Serious adverse events were reported in 49 (43%) of 115 patients in the ceritinib group and 36 (32%) of 113 in the chemotherapy group. Treatment-related serious adverse events were similar between groups (13 [11%] in the ceritinib group vs 12 [11%] in the chemotherapy group). The most frequent grade 3-4 adverse events in the ceritinib group were increased alanine aminotransferase concentration (24 [21%] of 115 vs two [2%] of 113 in the chemotherapy group), increased γ glutamyltransferase concentration (24 [21%] vs one [1%]), and increased aspartate aminotransferase concentration (16 [14%] vs one [1%] in the chemotherapy group). Six (5%) of 115 patients in the ceritinib group discontinued because of adverse events compared with eight (7%) of 116 in the chemotherapy group. 15 (13%) of 115 patients in the ceritinib group and five (4%) of 113 in the chemotherapy group died during the treatment period (from the day of the first dose of study treatment to 30 days after the final dose). 13 (87%) of the 15 patients who died in the ceritinib group died because of disease progression and two (13%) died because of an adverse event (one [7%] cerebrovascular accident and one [7%] respiratory failure); neither of these deaths were considered by the investigator to be treatment related. The five (4%) deaths in the chemotherapy group were all due to disease progression. These findings show that patients derive significant clinical benefit from a more potent ALK inhibitor after failure of crizotinib, and establish ceritinib as a more efficacious treatment option compared with chemotherapy in this patient population. Novartis Pharmaceuticals Corporation.


NHS dental professionals' evaluation of a child protection learning resource.
The aim of this survey was to evaluate the impact of an educational child protection resource which had been developed and made available, free of charge, to all NHS dental practices and salaried primary dental care services in England and concurrently published online. A postal questionnaire was sent to a random sample of 1000 NHS dental practices to assess whether the learning objectives of the educational resource had been met. A total of 467 questionnaires were completed (46.7% response rate). Almost two thirds of participants (63.4%) remembered receiving the Child Protection and Dental Team (CPDT) handbook or seeing the website and almost all of them had used (looked at or read) it and felt able to access it if needed. Of the 265 users, 76.2% felt it had improved their knowledge of child protection, 60.5% had adopted a child protection policy, 53.7% had identified a child protection lead and 25.8% had arranged further training as a result of using the educational resource. The findings from the evaluation indicated that the learning objectives of the CPDT educational resource had been met and highlighted ways in which the resource could be further improved to effectively meet the needs of dental professionals.


The protective effects of the supercritical-carbon dioxide fluid extract of Chrysanthemum indicum against lipopolysaccharide-induced acute lung injury in mice via modulating Toll-like receptor 4 signaling pathway.
The supercritical-carbon dioxide fluid extract of Chrysanthemum indicum Linné. (CFE) has been demonstrated to be effective in suppressing inflammation. The aim of this study is to investigate the preventive action and underlying mechanisms of CFE on acute lung injury (ALI) induced by lipopolysaccharide (LPS) in mice. ALI was induced by intratracheal instillation of LPS into lung, and dexamethasone was used as a positive control. Results revealed that pretreatment with CFE abated LPS-induced lung histopathologic changes, reduced the wet/dry ratio and proinflammatory cytokines productions (TNF-α, IL-1β, and IL-6), inhibited inflammatory cells migrations and protein leakages, suppressed the levels of MPO and MDA, and upregulated the abilities of antioxidative enzymes (SOD, CAT, and GPx). Furthermore, the pretreatment with CFE downregulated the activations of NF-κB and the expressions of TLR4/MyD88. These results suggested that CFE exerted potential protective effects against LPS-induced ALI in mice and was a potential therapeutic drug for ALI. Its mechanisms were at least partially associated with the modulations of TLR4 signaling pathways.


The effectiveness of acupuncture in the management of persistent regional myofascial head and neck pain: A systematic review and meta-analysis.
Persistent head and neck myofascial pain is among the most frequently reported pain complaints featuring major variability in treatment approaches and perception of improvement. Acupuncture is one of the least invasive complimentary modalities that can optimize conventional treatment. The aim of this review was to determine the evidence for the effectiveness of acupuncture in the management of localized persistent myofascial head and neck pain. Only randomized controlled clinical trials (RCTs) were included. The search was conducted in PubMed, Ovid Medline, Embase, Google Scholar, and Cochrane Library in addition to manual search. The main outcome measure was the comparison of the mean pain intensity score on VAS between acupuncture and sham-needling/no intervention groups. Safety data and adherence rate were also investigated. Six RCTs were identified with variable risk of bias. All included studies reported reduction in VAS pain intensity scores in the groups receiving acupuncture when compared to sham needling/no intervention. Meta-analysis, using a weighted mean difference as the effect estimate, included only 4 RCTs, revealed a 19.04 point difference in pain intensity between acupuncture and sham-needling/no intervention (95 %CI: -29.13 to -8.95). High levels of safety were demonstrated by the low rates of side effects/withdrawal. Inconsistency in reporting of outcomes was a major limitation. In conclusion, moderate-quality evidence suggests that acupuncture may be an effective and safe method in relieving persistent head and neck myofascial pain. Optimizing study designs and standardizing outcome measures are needed for future RCTs.


Effect of age and gender on serum periostin: Relationship to cortical measures, bone turnover and hormones.
Periostin is an extracellular matrix protein, and in bone is expressed most highly in the periosteum. It increases bone formation through osteoblast differentiation, cell adhesion, Wnt signalling and collagen cross-linking. We hypothesised that serum periostin would be high at times of life when cortical modeling is active, in early adulthood and in older age, and that it would correlate with cortical bone measures, bone turnover and hormones that regulate cortical modeling. We conducted a cross-sectional observational study of 166 healthy men and women at three skeletal stages; the end of longitudinal growth (16-18years), peak bone mass (30-32years) and older age (over 70years). We measured serum periostin with a new ELISA optimised for human serum and plasma which recognises all known splice variants (Biomedica). We measured the distal radius and distal tibia with HR-pQCT, and measured serum PINP, CTX, sclerostin, PTH, IGF-1, estradiol and testosterone. Periostin was higher at age 16-18 than age 30-32 (1253 vs 842pmol/l, p<0.001), but not different between age 30-32 and over age 70. Periostin was inversely correlated with tibia cortical thickness and density (R -0.229, -0.233, both p=0.003). It was positively correlated with PINP (R 0.529, p<0.001), CTX (R 0.427, p<0.001) and IGF-1 (R 0.440, p<0.001). When assessed within each age group these correlations were only significant at age 16-18, except for PINP which was also significant over age 70. We conclude that periostin may have a role in IGF-1 driven cortical modeling and consolidation in young adults, but it may not be an important mediator in older adults.


Large herbivores facilitate a dominant grassland forb via multiple indirect effects.
While large herbivores are critically important components of terrestrial ecosystems and can have pronounced top-down effects on plants, our understanding of the underlying mechanisms driving these effects remains incomplete. Large herbivores can alter plant growth, reproduction, and abundance through direct effects (predominantly consumption) and through indirect effects via altered interactions with abiotic factors and other species. We know considerably less about these indirect effects than the direct effects. Here, we integrate medium- and small-scale field experiments to investigate how a large vertebrate herbivore, cattle (Bos taurus), affects the aboveground biomass of a dominant forb species, Artemisia scoparia, via diverse direct and indirect pathways in a temperate grassland in northeast China. Although cattle consumed this forb, its biomass increased significantly in response to grazing, due to multiple indirect positive effects that outweighed the direct negative effects of consumption. Cattle preferentially consumed the competing grass Leymus chinensis, and altered Artemisia microhabitats by reducing total plant cover and litter biomass and by increasing the abundance of co-occurring ant species (e.g., Formica spp. and Lasius spp.). This led to additional indirect positive effects on A. scoparia likely due to (1) increased light availability in understory layers and other limiting resources (e.g., soil nutrients and moisture) caused by removal of competitors and plant litter at the soil surface and (2) the changes in resource availability (e.g., soil nutrients and moisture) associated with ant colonies. Our results show that large herbivores can affect plant growth not only via direct consumption, but also via multiple indirect effects. Focusing on the causes and consequences of herbivore-induced indirect effects will not only help us to better understand the influence of these animals in ecological systems, but will also lead to more effective land management and conservation practices in the regions they inhabit.


Accurate geometric characterization of gold nanorod ensemble by an inverse extinction/scattering spectroscopic method.
Aspect ratio, width, and end-cap factor are three critical parameters defined to characterize the geometry of metallic nanorod (NR). In our previous work [Opt. Express 21, 2987 (2013)], we reported an optical extinction spectroscopic (OES) method that can measure the aspect ratio distribution of gold NR ensembles effectively and statistically. However, the measurement accuracy was found to depend on the estimate of the width and end-cap factor of the nanorod, which unfortunately cannot be determined by the OES method itself. In this work, we propose to improve the accuracy of the OES method by applying an auxiliary scattering measurement of the NR ensemble which can help to estimate the mean width of the gold NRs effectively. This so-called optical extinction/scattering spectroscopic (OESS) method can fast characterize the aspect ratio distribution as well as the mean width of gold NR ensembles simultaneously. By comparing with the transmission electron microscopy experimentally, the OESS method shows the advantage of determining two of the three critical parameters of the NR ensembles (i.e., the aspect ratio and the mean width) more accurately and conveniently than the OES method.


Is climate change an unforeseen, irresistible and external factor - A force majeure in marine environmental law?
Several environmental laws include provisions on natural causes or force majeure, which except States from their commitments if it can be proven that the failure to meet the commitment is due to factors outside their control. The European Union Marine Strategy Framework Directive (MSFD) has a pivotal role in managing EU marine waters. This paper analyses natural causes and force majeure provisions of the MFSD and other marine legislation, and addresses their interaction with climate change and its consequences, especially the effect on the obligation of ensuring seas are in Good Environmental Status. Climate change is an exogenic unmanaged pressure in that it emanates from outside the area being managed but in which the management authority has to respond to the consequences of climate change, such as sea level rise and temperature elevation, rather than its causes. It is suggested that a defence by a Member State of force majeure may be accepted if an event was proven to be due to an externality of control, irresistible and unforeseeable. The analysis contends that countering such a legal defence would centre on the fact that climate change is a well-accepted phenomenon, is foreseen with an accepted level of confidence and probability and is due to human actions. However, as yet, this has not been legally tested.


Adiponectin: Serum-saliva associations and relations with oral and systemic markers of inflammation.
This study addresses gaps in our understanding about the validity and utility of using salivary adiponectin to index serum adiponectin levels. Matched blood and saliva samples were collected on a single occasion from healthy adults (n=99; age 18-36 years, 53% male). Serum and saliva was assayed for adiponectin and inflammatory cytokines (IL-1β, IL-6, IL-8, TNFα), and saliva was also assayed for markers of blood contamination (transferrin), total protein (salivary flow rate) and matrix metalloproteinase-8 (MMP-8). We examined the extent to which salivary adiponectin was associated with serum adiponectin, and the influence of potential confounders on the serum-saliva correlation, including age, sex, body mass index, and markers of inflammation, oral health, salivary blood contamination, and flow rate. Findings revealed a modest serum-saliva association for adiponectin, and strong positive associations between salivary adiponectin and salivary levels of inflammatory cytokines, MMP-8, transferrin, and total protein. By contrast, salivary adiponectin was not related to serum levels of inflammatory activity. The magnitude of the serum-saliva association was strengthened when controlling for total protein in saliva, blood leakage into oral fluid, salivary inflammatory cytokines, and MMP-8. The pattern of findings extends our understanding of salivary adiponectin and its potential use as an index of circulating adiponectin levels.


Metabolic efficiency with fast spiking in the squid axon.
Fundamentally, action potentials in the squid axon are consequence of the entrance of sodium ions during the depolarization of the rising phase of the spike mediated by the outflow of potassium ions during the hyperpolarization of the falling phase. Perfect metabolic efficiency with a minimum charge needed for the change in voltage during the action potential would confine sodium entry to the rising phase and potassium efflux to the falling phase. However, because sodium channels remain open to a significant extent during the falling phase, a certain overlap of inward and outward currents is observed. In this work we investigate the impact of ion overlap on the number of the adenosine triphosphate (ATP) molecules and energy cost required per action potential as a function of the temperature in a Hodgkin-Huxley model. Based on a recent approach to computing the energy cost of neuronal action potential generation not based on ion counting, we show that increased firing frequencies induced by higher temperatures imply more efficient use of sodium entry, and then a decrease in the metabolic energy cost required to restore the concentration gradients after an action potential. Also, we determine values of sodium conductance at which the hydrolysis efficiency presents a clear minimum.


Evidence for a (15)N positive excursion in terrestrial foodwebs at the Middle to Upper Palaeolithic transition in south-western France: Implications for early modern human palaeodiet and palaeoenvironment.
The Middle to Upper Palaeolithic transition around 35,000 years ago coincides with the replacement of Neanderthals by anatomically modern humans in Europe. Several hypotheses have been suggested to explain this replacement, one of them being the ability of anatomically modern humans to broaden their dietary spectrum beyond the large ungulate prey that Neanderthals consumed exclusively. This scenario is notably based on higher nitrogen-15 amounts in early Upper Palaeolithic anatomically modern human bone collagen compared with late Neanderthals. In this paper, we document a clear increase of nitrogen-15 in bone collagen of terrestrial herbivores during the early Aurignacian associated with anatomically modern humans compared with the stratigraphically older Châtelperronian and late Mousterian fauna associated with Neanderthals. Carnivores such as wolves also exhibit a significant increase in nitrogen-15, which is similar to that documented for early anatomically modern humans compared with Neanderthals in Europe. A shift in nitrogen-15 at the base of the terrestrial foodweb is responsible for such a pattern, with a preserved foodweb structure before and after the Middle to Upper Palaeolithic transition in south-western France. Such an isotopic shift in the terrestrial ecosystem may be due to an increase in aridity during the time of deposition of the early Aurignacian layers. If it occurred across Europe, such a shift in nitrogen-15 in terrestrial foodwebs would be enough to explain the observed isotopic trend between late Neanderthals and early anatomically modern humans, without any significant change in the diet composition at the Middle to Upper Palaeolithic transition.


Substrate deformation levels associated with routine physical activity are less stimulatory to bone cells relative to loading-induced oscillatory fluid flow.
Although it is well accepted that bone tissue metabolism is regulated by external mechanical loads, it remains unclear to what load-induced physical signals bone cells respond. In this study, a novel computer-controlled stretch device and parallel plate flow chamber were employed to investigate cytosolic calcium (Ca2+i) mobilization in response to a range of dynamic substrate strain levels (0.1-10 percent, 1 Hz) and oscillating fluid flow (2 N/m2, 1 Hz). In addition, we quantified the effect of dynamic substrate strain and oscillating fluid flow on the expression of mRNA for the bone matrix protein osteopontin (OPN). Our data demonstrate that continuum strain levels observed for routine physical activities (< 0.5 percent) do not induce Ca2+i responses in osteoblastic cells in vitro. However, there was a significant increase in the number of responding cells at larger strain levels. Moreover, we found no change in osteopontin mRNA level in response to 0.5 percent strain at 1 Hz. In contrast, oscillating fluid flow predicted to occur in the lacunar-canalicular system due to routine physical activities (2 N/m2, 1 Hz) caused significant increases in both Ca2+i and OPN mRNA. These data suggest that, relative to fluid flow, substrate deformation may play less of a role in bone cell mechanotransduction associated with bone adaptation to routine loads.


Drought severity and all-cause mortality rates among adults in the United States: 1968-2014.
Little is known about the effect of drought on all-cause mortality, especially in higher income countries such as the United States. As the frequency and severity of droughts are likely to increase, understanding the connections between drought and mortality becomes increasingly important. Our exposure variable was an annual cumulative drought severity score based on the 1-month, county-level Standardized Precipitation Evapotranspiration Index. The outcome variables of demographic subgroup-specific all-cause mortality count data per year were obtained from the National Vital Statistics System. Any counts below 10 deaths were censored in that demographic group per county. We modeled county-stratum-year mortality using interval-censored negative binomial regression with county-level random intercepts, for each combined age-race-sex stratum either with or without further stratification by climate regions. Fixed effects meta-regression was used to test the associations between age, race, sex, and region with the drought-mortality regression coefficients. Predictive margins were then calculated from the meta-regression model to estimate larger subgroup (e.g., 'race' or 'sex') associations of drought with mortality. Most of the results were null for associations between drought severity and mortality, across joint strata of race, age, sex and region, but incidence rate ratios (IRRs) for 17 subgroups were significant after accounting for the multiple testing; ten were < 1 indicating a possible protective effect of drought on mortality for that particular subpopulation. The meta-regression indicated heterogeneity in the association of drought with mortality according to race, climate region, and age, but not by sex. Marginal means of the estimated log-incidence rate ratios differed significantly from zero for age groups 25-34, 35-44, 45-54 and 55-64; for the white race group; and for the South, West and Southwest regions, in the analysis that included wet county-years. The margin of the meta-regression model suggested a slightly negative, but not statistically significant, association of drought with same-year mortality in the overall population. There were significant, heterogeneous-direction associations in subpopulation-stratified models, after controlling for multiple comparisons, suggesting that the impacts of drought on mortality may not be monolithic across the United States. Meta-regression identified systematic differences in the associations of drought severity with all-cause mortality according to climate region, race, and age. These findings suggest there may be important contextual differences in the effects of drought severity on mortality, motivating further work focused on local mechanisms. We speculate that some of the estimated negative associations of drought severity with same-year mortality could be consistent with either a protective effect of drought on total mortality in the same year, or with a delayed health effect of drought beyond the same year. Further research is needed to clarify associations of drought with more specific causes of death and with sublethal health outcomes, for specific subpopulations, and considering lagged effects occurring beyond the same year as the drought.


PD-L1 on tumor cells is induced in ascites and promotes peritoneal dissemination of ovarian cancer through CTL dysfunction.
Ovarian cancer often progresses by disseminating to the peritoneal cavity, but how the tumor cells evade host immunity during this process is poorly understood. Programmed cell death 1 ligand 1 (PD-L1) is known to suppress immune system and to be expressed in cancer cells. The purpose of this study is to elucidate the function of PD-L1 in peritoneal dissemination. Ovarian cancer cases were studied by microarray and immunohistochemistry. PD-L1 expression in mouse ovarian cancer cell line in various conditions was assessed by flow cytometry. PD-L1-overexpression cell line and PD-L1-depleted cell line were generated, and cytolysis by CTLs was analyzed, and alterations in CTLs were studied by means of timelapse and microarray. These cell lines were injected intraperitoneally to syngeneic immunocompetent mice. Microarray and immunohistochemistry in human ovarian cancer revealed significant correlation between PD-L1 expression and peritoneal positive cytology. PD-L1 expression in mouse ovarian cancer cells was induced upon encountering lymphocytes in the course of peritoneal spread in vivo and coculture with lymphocytes in vitro. Tumor cell lysis by CTLs was attenuated when PD-L1 was overexpressed and promoted when it was silenced. PD-L1 overexpression inhibited gathering and degranulation of CTLs. Gene expression profile of CTLs caused by PD-L1-overexpressing ovarian cancer was associated with CTLs exhaustion. In mouse models, PD-L1 depletion resulted in inhibited tumor growth in the peritoneal cavity and prolonged survival. PD-L1 expression in tumor cell promotes peritoneal dissemination by repressing CTL function. PD-L1-targeted therapy is a promising strategy for preventing and treating peritoneal dissemination.


Haemolytic-uraemic syndrome: clinical experience of an outbreak in the West Midlands.
In 1982-3, 35 children from the West Midlands developed the haemolytic-uraemic syndrome. This was a higher incidence than expected and included an epidemic localised to the Wolverhampton area in July 1983 which comprised 11 cases in two weeks. Twenty three children were treated with dialysis, of whom three died. Six patients developed chronic renal failure, four of them from Wolverhampton. Extrarenal manifestations included neurological sequelae in four, two of whom also developed insulin dependent diabetes mellitus and chronic renal failure. Cardiomyopathy occurred in one child, who also had chronic renal failure. The outcome of these 35 patients was not predictable from prognostic criteria derived from previous experience in Britain. This, together with the high prevalence of extrarenal disease and the geographical localisation of the 1983 outbreak, suggested an aetiological agent new to the region. Faeces from 10 patients were examined for verotoxin producing Escherichia coli, and positive strains of serotype O157.H7 were found in three patients during the Wolverhampton outbreak.


A matter of time: improvement of visual temporal processing during training-induced restoration of light detection performance.
The issue of how basic sensory and temporal processing are related is still unresolved. We studied temporal processing, as assessed by simple visual reaction times (RT) and double-pulse resolution (DPR), in patients with partial vision loss after visual pathway lesions and investigated whether vision restoration training (VRT), a training program designed to improve light detection performance, would also affect temporal processing. Perimetric and campimetric visual field tests as well as maps of DPR thresholds and RT were acquired before and after a 3 months training period with VRT. Patient performance was compared to that of age-matched healthy subjects. Intact visual field size increased during training. Averaged across the entire visual field, DPR remained constant while RT improved slightly. However, in transition zones between the blind and intact areas (areas of residual vision) where patients had shown between 20 and 80% of stimulus detection probability in pre-training visual field tests, both DPR and RT improved markedly. The magnitude of improvement depended on the defect depth (or degree of intactness) of the respective region at baseline. Inter-individual training outcome variability was very high, with some patients showing little change and others showing performance approaching that of healthy controls. Training-induced improvement of light detection in patients with visual field loss thus generalized to dynamic visual functions. The findings suggest that similar neural mechanisms may underlie the impairment and subsequent training-induced functional recovery of both light detection and temporal processing.


Classification and Weakly Supervised Pain Localization using Multiple Segment Representation.
Automatic pain recognition from videos is a vital clinical application and, owing to its spontaneous nature, poses interesting challenges to automatic facial expression recognition (AFER) research. Previous pain vs no-pain systems have highlighted two major challenges: (1) ground truth is provided for the sequence, but the presence or absence of the target expression for a given frame is unknown, and (2) the time point and the duration of the pain expression event(s) in each video are unknown. To address these issues we propose a novel framework (referred to as MS-MIL) where each sequence is represented as a bag containing multiple segments, and multiple instance learning (MIL) is employed to handle this weakly labeled data in the form of sequence level ground-truth. These segments are generated via multiple clustering of a sequence or running a multi-scale temporal scanning window, and are represented using a state-of-the-art Bag of Words (BoW) representation. This work extends the idea of detecting facial expressions through 'concept frames' to 'concept segments' and argues through extensive experiments that algorithms such as MIL are needed to reap the benefits of such representation. The key advantages of our approach are: (1) joint detection and localization of painful frames using only sequence-level ground-truth, (2) incorporation of temporal dynamics by representing the data not as individual frames but as segments, and (3) extraction of multiple segments, which is well suited to signals with uncertain temporal location and duration in the video. Extensive experiments on UNBC-McMaster Shoulder Pain dataset highlight the effectiveness of the approach by achieving competitive results on both tasks of pain classification and localization in videos. We also empirically evaluate the contributions of different components of MS-MIL. The paper also includes the visualization of discriminative facial patches, important for pain detection, as discovered by our algorithm and relates them to Action Units that have been associated with pain expression. We conclude the paper by demonstrating that MS-MIL yields a significant improvement on another spontaneous facial expression dataset, the FEEDTUM dataset.


RNA-seq analysis identifies an intricate regulatory network controlling cluster root development in white lupin.
Highly adapted plant species are able to alter their root architecture to improve nutrient uptake and thrive in environments with limited nutrient supply. Cluster roots (CRs) are specialised structures of dense lateral roots formed by several plant species for the effective mining of nutrient rich soil patches through a combination of increased surface area and exudation of carboxylates. White lupin is becoming a model-species allowing for the discovery of gene networks involved in CR development. A greater understanding of the underlying molecular mechanisms driving these developmental processes is important for the generation of smarter plants for a world with diminishing resources to improve food security. RNA-seq analyses for three developmental stages of the CR formed under phosphorus-limited conditions and two of non-cluster roots have been performed for white lupin. In total 133,045,174 high-quality paired-end reads were used for a de novo assembly of the root transcriptome and merged with LAGI01 (Lupinus albus gene index) to generate an improved LAGI02 with 65,097 functionally annotated contigs. This was followed by comparative gene expression analysis. We show marked differences in the transcriptional response across the various cluster root stages to adjust to phosphate limitation by increasing uptake capacity and adjusting metabolic pathways. Several transcription factors such as PLT, SCR, PHB, PHV or AUX/IAA with a known role in the control of meristem activity and developmental processes show an increased expression in the tip of the CR. Genes involved in hormonal responses (PIN, LAX, YUC) and cell cycle control (CYCA/B, CDK) are also differentially expressed. In addition, we identify primary transcripts of miRNAs with established function in the root meristem. Our gene expression analysis shows an intricate network of transcription factors and plant hormones controlling CR initiation and formation. In addition, functional differences between the different CR developmental stages in the acclimation to phosphorus starvation have been identified.


Prediction of amyloid pathology in cognitively unimpaired individuals using voxel-wise analysis of longitudinal structural brain MRI.
Magnetic resonance imaging (MRI) has unveiled specific alterations at different stages of Alzheimer's disease (AD) pathophysiologic continuum constituting what has been established as "AD signature". To what extent MRI can detect amyloid-related cerebral changes from structural MRI in cognitively unimpaired individuals is still an area open for exploration. Longitudinal 3D-T1 MRI scans were acquired from a subset of the ADNI cohort comprising 403 subjects: 79 controls (Ctrls), 50 preclinical AD (PreAD), and 274 MCI and dementia due to AD (MCI/AD). Amyloid CSF was used as gold-standard measure with established cutoffs (< 192 pg/mL) to establish diagnostic categories. Cognitively unimpaired individuals were defined as Ctrls if were amyloid negative and PreAD otherwise. The MCI/AD group was amyloid positive. Only subjects with the same diagnostic category at baseline and follow-up visits were considered for the study. Longitudinal morphometric analysis was performed using SPM12 to calculate Jacobian determinant maps. Statistical analysis was carried out on these Jacobian maps to identify structural changes that were significantly different between diagnostic categories. A machine learning classifier was applied on Jacobian determinant maps to predict the presence of abnormal amyloid levels in cognitively unimpaired individuals. The performance of this classifier was evaluated using receiver operating characteristic curve analysis and as a function of the follow-up time between MRI scans. We applied a cost function to assess the benefit of using this classifier in the triaging of individuals in a clinical trial-recruitment setting. The optimal follow-up time for classification of Ctrls vs PreAD was Δt > 2.5 years, and hence, only subjects within this temporal span are used for evaluation (15 Ctrls, 10 PreAD). The longitudinal voxel-based classifier achieved an AUC = 0.87 (95%CI 0.72-0.97). The brain regions that showed the highest discriminative power to detect amyloid abnormalities were the medial, inferior, and lateral temporal lobes; precuneus; caudate heads; basal forebrain; and lateral ventricles. Our work supports that machine learning applied to longitudinal brain volumetric changes can be used to predict, with high precision, the presence of amyloid abnormalities in cognitively unimpaired subjects. Used as a triaging method to identify a fixed number of amyloid-positive individuals, this longitudinal voxel-wise classifier is expected to avoid 55% of unnecessary CSF and/or PET scans and reduce economic cost by 40%.