Increased methane emissions from oil and gas following the Soviet Union's collapse.

Global atmospheric methane concentrations rose by 10 to 15 ppb/y in the 1980s before abruptly slowing to 2 to 8 ppb/y in the early 1990s. This period in the 1990s is known as the "methane slowdown" and has been attributed in part to the collapse of the former Soviet Union (USSR) in December 1991, which may have decreased the methane emissions from oil and gas operations. Here, we develop a methane plume detection system based on probabilistic deep learning and human-labeled training data. We use this method to detect methane plumes from Landsat 5 satellite observations over Turkmenistan from 1986 to 2011. We focus on Turkmenistan because economic data suggest it could account for half of the decline in oil and gas emissions from the former USSR. We find an increase in both the frequency of methane plume detections and the magnitude of methane emissions following the collapse of the USSR. We estimate a national loss rate from oil and gas infrastructure in Turkmenistan of more than 10% at times, which suggests the socioeconomic turmoil led to a lack of oversight and widespread infrastructure failure in the oil and gas sector. Our finding of increased oil and gas methane emissions from Turkmenistan following the USSR's collapse casts doubt on the long-standing hypothesis regarding the methane slowdown, begging the question: "what drove the 1992 methane slowdown?"

Targeting ON-bipolar cells by AAV gene therapy stably reverses LRIT3-congenital stationary night blindness.

SignificanceCanine models of inherited retinal diseases have helped advance adeno-associated virus (AAV)-based gene therapies targeting specific cells in the outer retina for treating blinding diseases in patients. However, therapeutic targeting of diseases such as congenital stationary night blindness (CSNB) that exhibit defects in ON-bipolar cells (ON-BCs) of the midretina remains underdeveloped. Using a leucine-rich repeat, immunoglobulin-like and transmembrane domain 3 (LRIT3) mutant canine model of CSNB exhibiting ON-BC dysfunction, we tested the ability of cell-specific AAV capsids and promotors to specifically target ON-BCs for gene delivery. Subretinal injection of one vector demonstrated safety and efficacy with robust and stable rescue of electroretinography signals and night vision up to 1 y, paving the way for clinical trials in patients.

Techniques for subretinal injections in animals.

Subretinal injections are not commonly performed during clinical treatment of animals but are frequently used in laboratory animal models to assess therapeutic efficacy and safety of gene and cell therapy products. Veterinary ophthalmologists are often employed to perform the injections in the laboratory animal setting, due to knowledge of comparative ocular anatomy between species and familiarity with operating on non-human eyes. Understanding the different approaches used for subretinal injection in each species and potential complications that may be encountered is vital to achieving successful and reproducible results. This manuscript provides a summary of different approaches to subretinal injections in the most common animal model species, along with information from published literature and experience of the authors to educate novice or experienced surgeons tasked with performing these injections for the first time.

Exploring 3D leaf anatomical traits for C4 photosynthesis: chloroplast and plasmodesmata pit field size in maize and sugarcane.

Volume and surface area of chloroplasts and surface area of plasmodesmata pit fields are presented for two C4 species, maize and sugarcane, with respect to cell surface area and cell volume. Serial block face scanning electron microscopy (SBF-SEM) and confocal laser scanning microscopy with the Airyscan system (LSM) were used. Chloroplast size estimates were much faster and easier using LSM than with SBF-SEM; however, the results were more variable than SBF-SEM. Mesophyll cells were lobed where chloroplasts were located, facilitating cell-to-cell connections while allowing for greater intercellular airspace exposure. Bundle sheath cells were cylindrical with chloroplasts arranged centrifugally. Chloroplasts occupied c. 30-50% of mesophyll cell volume, and 60-70% of bundle sheath cell volume. Roughly 2-3% of each cell surface area was covered by plasmodesmata pit fields for both bundle sheath and mesophyll cells. This work will aid future research to develop SBF-SEM methodologies with the aim to better understand the effect of cell structure on C4 photosynthesis.

Characterization and computational simulation of human Syx, a RhoGEF implicated in glioblastoma.

Structural discovery of guanine nucleotide exchange factor (GEF) protein complexes is likely to become increasingly relevant with the development of new therapeutics targeting small GTPases and development of new classes of small molecules that inhibit protein-protein interactions. Syx (also known as PLEKHG5 in humans) is a RhoA GEF implicated in the pathology of glioblastoma (GBM). Here we investigated protein expression and purification of ten different human Syx constructs and performed biophysical characterizations and computational studies that provide insights into why expression of this protein was previously intractable. We show that human Syx can be expressed and isolated and Syx is folded as observed by circular dichroism (CD) spectroscopy and actively binds to RhoA as determined by co-elution during size exclusion chromatography (SEC). This characterization may provide critical insights into the expression and purification of other recalcitrant members of the large class of oncogenic-Diffuse B-cell lymphoma (Dbl) homology GEF proteins. In addition, we performed detailed homology modeling and molecular dynamics simulations on the surface of a physiologically realistic membrane. These simulations reveal novel insights into GEF activity and allosteric modulation by the plekstrin homology (PH) domain. These newly revealed interactions between the GEF PH domain and the membrane embedded region of RhoA support previously unexplained experimental findings regarding the allosteric effects of the PH domain from numerous activity studies of Dbl homology GEF proteins. This work establishes new hypotheses for structural interactivity and allosteric signal modulation in Dbl homology RhoGEFs.

Effects of Altering HSPG Binding and Capsid Hydrophilicity on Retinal Transduction by AAV.

Adeno-associated viruses (AAVs) have recently emerged as the leading vector for retinal gene therapy. However, AAV vectors which are capable of achieving clinically relevant levels of transgene expression and widespread retinal transduction are still an unmet need. Using rationally designed AAV2-based capsid variants, we investigate the role of capsid hydrophilicity and hydrophobicity as it relates to retinal transduction. We show that hydrophilic, single amino acid (aa) mutations (V387R, W502H, E530K, L583R) in AAV2 negatively impact retinal transduction when heparan sulfate proteoglycan (HSPG) binding remains intact. Conversely, addition of hydrophobic point mutations to an HSPG binding deficient capsid (AAV2ΔHS) lead to increased retinal transduction in both mouse and macaque. Our top performing vector, AAV2(4pMut)ΔHS, achieved robust rod and cone photoreceptor (PR) transduction in macaque, especially in the fovea, and demonstrates the ability to spread laterally beyond the borders of the subretinal injection (SRI) bleb. This study both evaluates biophysical properties of AAV capsids that influence retinal transduction, and assesses the transduction and tropism of a novel capsid variant in a clinically relevant animal model.ImportanceRationally guided engineering of AAV capsids aims to create new generations of vectors with enhanced potential for human gene therapy. By applying rational design principles to AAV2-based capsids, we evaluated the influence of hydrophilic and hydrophobic amino acid (aa) mutations on retinal transduction as it relates to vector administration route. Through this approach we identified a largely deleterious relationship between hydrophilic aa mutations and canonical HSPG binding by AAV2-based capsids. Conversely, the inclusion of hydrophobic aa substitutions on a HSPG binding deficient capsid (AAV2ΔHS), generated a vector capable of robust rod and cone photoreceptor (PR) transduction. This vector AAV2(4pMut)ΔHS also demonstrates a remarkable ability to spread laterally beyond the initial subretinal injection (SRI) bleb, making it an ideal candidate for the treatment of retinal diseases which require a large area of transduction.

Increased bundle-sheath leakiness of CO2 during photosynthetic induction shows a lack of coordination between the C4 and C3 cycles.

Use of a complete dynamic model of NADP-malic enzyme C4 photosynthesis indicated that, during transitions from dark or shade to high light, induction of the C4 pathway was more rapid than that of C3 , resulting in a predicted transient increase in bundle-sheath CO2 leakiness (ϕ). Previously, ϕ has been measured at steady state; here we developed a new method, coupling a tunable diode laser absorption spectroscope with a gas-exchange system to track ϕ in sorghum and maize through the nonsteady-state condition of photosynthetic induction. In both species, ϕ showed a transient increase to > 0.35 before declining to a steady state of 0.2 by 1500 s after illumination. Average ϕ was 60% higher than at steady state over the first 600 s of induction and 30% higher over the first 1500 s. The transient increase in ϕ, which was consistent with model prediction, indicated that capacity to assimilate CO2 into the C3 cycle in the bundle sheath failed to keep pace with the rate of dicarboxylate delivery by the C4 cycle. Because nonsteady-state light conditions are the norm in field canopies, the results suggest that ϕ in these major crops in the field is significantly higher and energy conversion efficiency lower than previous measured values under steady-state conditions.

Sterols in an intramolecular channel of Smoothened mediate Hedgehog signaling.

Smoothened (SMO), a class Frizzled G protein-coupled receptor (class F GPCR), transduces the Hedgehog signal across the cell membrane. Sterols can bind to its extracellular cysteine-rich domain (CRD) and to several sites in the seven transmembrane helices (7-TMs) of SMO. However, the mechanism by which sterols regulate SMO via multiple sites is unknown. Here we determined the structures of SMO-Gi complexes bound to the synthetic SMO agonist (SAG) and to 24(S),25-epoxycholesterol (24(S),25-EC). A novel sterol-binding site in the extracellular extension of TM6 was revealed to connect other sites in 7-TMs and CRD, forming an intramolecular sterol channel from the middle side of 7-TMs to CRD. Additional structures of two gain-of-function variants, SMOD384R and SMOG111C/I496C, showed that blocking the channel at its midpoints allows sterols to occupy the binding sites in 7-TMs, thereby activating SMO. These data indicate that sterol transport through the core of SMO is a major regulator of SMO-mediated signaling.

Atypical chorioretinal lesions in Siberian Husky dogs with primary angle-closure glaucoma: a case series.

BACKGROUND: A number of etiologies for different canine chorioretinal lesions have been proved or suggested but some fundic lesions remain unclear in terms of an etiologic diagnosis, treatment options and prognosis. The purpose of this case series is to describe atypical chorioretinal lesions observed in dogs with primary angle-closure glaucoma (PACG). CASE PRESENTATION: Two spayed-female Siberian Huskies (3- and 4-year-old) and one Siberian Husky/Australian Shepherd mixed breed dog (11-month-old) that had multifocal depigmented retinal lesions and PACG were included. PROCEDURES: Ophthalmic examination, gross, and histopathologic examination findings are described. One of the dogs underwent further clinical diagnostics. Advanced clinical diagnostics on the fellow, presumed to be non-glaucomatous eye of a dog revealed: pectinate ligament dysplasia by gonioscopy, retinal thinning in the depigmented area and wedge shaped retinal thinning with delayed choroidal vascular perfusion by optical coherence tomography, confocal scanning laser ophthalmoscopy, fluorescein and indocyanine green angiography. Quantifiable maze testing for the same eye revealed mild nyctalopia but the full-field electroretinogram showed no generalized decrease of retinal function. Genetic testing for mutations within the retinitis pigmentosa GTPase regulator gene causing X-linked progressive retinal atrophy in Siberian Huskies was negative. Histopathologic evaluations on enucleated eyes in two dogs confirmed goniodysgenesis, PACG with optic nerve head cupping, and diffuse inner retinal atrophy. In addition, segmental profound retinal atrophy, loss of retinal pigment epithelium, and adhesion of the retina to Bruch's membrane was observed and coincided with multifocal depigmented lesions noted on fundic examination. CONCLUSIONS: To our knowledge, this is the first case series with clinical and histopathologic data of chorioretinal lesions, most likely caused by severely impaired choroidal perfusion. Further studies are warranted to elucidate the etiology and pathophysiology, including its possible association with PACG.

'Against the cult of veganism': Unpacking the social psychology and ideology of anti-vegans.

Despite the established health and ecological benefits of a plant-based diet, the decision to eschew meat and other animal-derived food products remains controversial. So polarising is this topic that anti-vegan communities - groups of individuals who stand vehemently against veganism - have sprung up across the internet. Much scholarship on veganism characterizes anti-vegans in passing, painting them as ill-informed, uneducated, or simply obstinate. However, little empirical work has investigated these communities and the individuals within them. Accordingly, we conducted a study using social media data from the popular platform, Reddit. Specifically, we collected all available submissions (∼3523) and comments (∼45,528) from r/AntiVegan subreddit users (N = 3819) over a five-year period. Using a battery of computerized text analytic tools, we examined the psychosocial characteristics of Reddit users who publicly identify as anti-vegan, how r/AntiVegan users discuss their beliefs, and how the individual user changes as a function of community membership. Results from our analyses suggest several individual differences that align r/AntiVegan users with the community, including dark entertainment, ex-veganism and science denial. Several topics were extensively discussed by r/AntiVegan members, including nuanced discourse on the ethicality and health implications of vegan diets, and the naturalness of animal death, which ran counter to our expectations and lay stereotypes of r/AntiVegan users. Finally, several longitudinal changes in language use were observed within the community, reflecting enhanced group commitment over time, including an increase in group-focused language and a decrease in cognitive processing. Implications for vegan-nonvegan relations are discussed.

Examining long-term trends in politics and culture through language of political leaders and cultural institutions.

From many perspectives, the election of Donald Trump was seen as a departure from long-standing political norms. An analysis of Trump's word use in the presidential debates and speeches indicated that he was exceptionally informal but at the same time, spoke with a sense of certainty. Indeed, he is lower in analytic thinking and higher in confidence than almost any previous American president. Closer analyses of linguistic trends of presidential language indicate that Trump's language is consistent with long-term linear trends, demonstrating that he is not as much an outlier as he initially seems. Across multiple corpora from the American presidents, non-US leaders, and legislative bodies spanning decades, there has been a general decline in analytic thinking and a rise in confidence in most political contexts, with the largest and most consistent changes found in the American presidency. The results suggest that certain aspects of the language style of Donald Trump and other recent leaders reflect long-evolving political trends. Implications of the changing nature of popular elections and the role of media are discussed.

Emotion regulation across the lifespan: age differences in intrapersonal and interpersonal strategies for the adjustment to the COVID-19 pandemic in four countries.

OBJECTIVES: Studies have shown age differences in adapting to the COVID-19 pandemic. The processes explaining these age differences remain unclear. Intrapersonal and interpersonal emotion regulation play an important role in psycho-social adjustment and develop across the lifespan. This study investigated whether differences in COVID-19-related adjustment disorder symptoms can be explained by age-differences in rumination in a multi-national sample. Furthermore, linguistic indicators of ruminative processing were examined with reference to age. METHODS: N = 1401 participants (from USA, UK, Switzerland, Germany, aged 18-88) completed an online survey and a writing task. Measures included brooding, co-brooding, adjustment disorder symptoms, and language indicators of negative self- and communal-focus . RESULTS: Older participants reported less adjustment disorder symptoms which was mediated by less (co-)brooding. Participants reporting more (co-)brooding wrote about COVID-19 more negatively. While in younger adults more self-focus was associated with higher ruminative brooding, in older adults it was associated with less brooding. CONCLUSION: These findings contribute to a better understanding of regulatory mechanisms that help explain age differences in mental health. They warrant further research considering age-related differences, as our results suggest not only more adaptive emotion regulation as resilience factor in older individuals but also different qualities of self-focus while processing stressful events.

Use of open-ended questionnaires to examine the effects of tinnitus and its relation to patient-reported outcome measures.

OBJECTIVE: The primary aim of the study was to examine the automated linguistic analysis of the open-ended problem (PQ) and life-effects (LEQ) questionnaires to understand the psychological effects of tinnitus. DESIGN: The study used a cross-sectional design. Participants completed online questionnaires which included demographic questions, several standardised patient-reported outcome measures (PROMs), and two open-ended questions focussing on PQ and LEQ related to tinnitus. The response to open-ended questions was analysed using the Linguistic Inquiry Word Count (LIWC) software to identify the frequency of text on various linguistic dimensions relevant to tinnitus. STUDY SAMPLE: 336 individuals with tinnitus. RESULTS: The study results point to two broad findings. First, although PQ and LEQ have some similarities with PROMs (e.g. the linguistic dimension negative emotions having a weak positive correlation with anxiety and depression), no correlation with the number of dimensions suggests that the open-ended questions identify additional elements that are not captured in PROMs. Second, more linguistic dimensions from the PQ correlate with PROMs compared to LEQ suggesting that the current PROMs are problem-oriented. CONCLUSIONS: The study results support the idea that the use of open-ended questions in addition to PROMs may help optimise the efforts in examining the effects of chronic conditions such as tinnitus.

Correlations Between Retinal Optical Coherence Tomography and Histopathology in Preclinical Safety Assessment of Ocular Therapies.

METHODS: Twelve studies that involved intravitreal or subretinal test article administration in multiple laboratory animal species and employed both OCT and histopathologic assessment were identified. These data were compared to determine the degree of correlation for each identified abnormality. RESULTS: Severity of observed OCT and histopathological changes ranged from minimal to severe, with moderate and severe changes having a higher rate of agreement between the 2 techniques. Changes to well-defined structures, including the retinal blood vessels, optic nerve, and retinal pigment epithelium, also showed a strong correlation. CONCLUSIONS: There was a strong correlation between OCT and histopathology in both intravitreal and subretinal injection studies, demonstrating the value of collaboration between the study ophthalmologist and pathologist and offering translatable means to monitor pharmacological or toxicological effects in preclinical toxicological studies.

Novel AAV44.9-Based Vectors Display Exceptional Characteristics for Retinal Gene Therapy.

The majority of inherited retinal diseases (IRDs) are caused by mutations in genes expressed in photoreceptors (PRs). The ideal vector to address these conditions is one that transduces PRs in large areas of retina with the smallest volume/lowest titer possible, and efficiently transduces foveal cones, the cells responsible for acute, daylight vision that are often the only remaining area of functional retina in IRDs. The purpose of our study was to evaluate the retinal tropism and potency of a novel capsid, AAV44.9, and rationally designed derivatives thereof. We found that AAV44.9 and AAV44.9(E531D) transduced retinas of subretinally injected (SRI) mice with higher efficiency than did benchmark AAV5- and AAV8-based vectors. In macaques, highly efficient cone and rod transduction was observed following submacular and peripheral SRI. AAV44.9- and AAV44.9(E531D)-mediated GFP fluorescence extended laterally well beyond SRI bleb margins. Notably, extrafoveal injection (i.e., fovea not detached during surgery) led to transduction of up to 98% of foveal cones. AAV44.9(E531D) efficiently transduced parafoveal and perifoveal cones, whereas AAV44.9 did not. AAV44.9(E531D) was also capable of restoring retinal function to a mouse model of IRD. These novel capsids will be useful for addressing IRDs that would benefit from an expansive treatment area.

Dirty laundry: The nature and substance of seeking relationship help from strangers online.

Interpersonal relationships are vital to our well-being. In recent years, it has become increasingly common to seek relationship help through anonymous online platforms. Accordingly, we conducted a large-scale analysis of real-world relationship help-seeking to create a descriptive overview of the nature and substance of online relationship help-seeking. By analyzing the demographic characteristics and language of relationship help-seekers on Reddit (N = 184,631), we establish the first-ever big data analysis of relationship help-seeking and relationship problems in situ among the general population. Our analyses highlight real-world relationship struggles found in the general population, extending beyond past work that is typically limited to counseling/intervention settings. We find that relationship problem estimates from our sample are closer to those found in the general population, providing a more generalized insight into the distribution and prevalence of relationship problems as compared with past work. Further, we find several meaningful associations between relationship help-seeking behavior, gender, and attachment. Notably, numerous gender differences in help-seeking and romantic attachment emerged. Our findings suggest that, contrary to more traditional contexts, men are more likely to seek help with their relationships online, are more expressive of their emotions (e.g., discussing the topic of "heartache"), and show language patterns generally consistent with more secure attachment. Our analyses highlight pathways for further exploration, providing even deeper insights into the timing, lifecycle, and moderating factors that influence who, what, why, and how people seek help for their interpersonal relationships.

Natural Language Analysis and the Psychology of Verbal Behavior: The Past, Present, and Future States of the Field.

Throughout history, scholars and laypeople alike have believed that our words contain subtle clues about what we are like as people, psychologically speaking. However, the ways in which language has been used to infer psychological processes has seen dramatic shifts over time and, with modern computational technologies and digital data sources, we are on the verge of a massive revolution in language analysis research. In this article, we discuss the past and current states of research at the intersection of language analysis and psychology, summarizing the central successes and shortcomings of psychological text analysis to date. We additionally outline and discuss a critical need for language analysis practitioners in the social sciences to expand their view of verbal behavior. Lastly, we discuss the trajectory of interdisciplinary research on language and the challenges of integrating analysis methods across paradigms, recommending promising future directions for the field along the way.

Temperature response of Rubisco kinetics in Arabidopsis thaliana: thermal breakpoints and implications for reaction mechanisms.

Enhancement of Rubisco kinetics could improve photosynthetic efficiency, ultimately resulting in increased crop yield. However, imprecise knowledge of the reaction mechanism and the individual rate constants limits our ability to optimize the enzyme. Membrane inlet mass spectrometry (MIMS) may offer benefits over traditional methods for determining individual rate constants of the Rubisco reaction mechanism, as it can directly monitor concentration changes in CO2, O2, and their isotopologs during assays. However, a direct comparison of MIMS with the traditional radiolabel method of determining Rubisco kinetic parameters has not been made. Here, the temperature responses of Rubisco kinetic parameters from Arabidopsis thaliana were measured using radiolabel and MIMS methods. The two methods provided comparable parameters above 25 °C, but temperature responses deviated at low temperature as MIMS-derived catalytic rates of carboxylation, oxygenation, and CO2/O2 specificity showed thermal breakpoints. Here, we discuss the variability and uncertainty surrounding breakpoints in the Rubisco temperature response and the relevance of individual rate constants of the reaction mechanisms to potential breakpoints.

Phenotype of macular corneal dystrophy in Labrador Retrievers: A multicenter study.

OBJECTIVE: To describe the phenotype of canine macular corneal dystrophy (MCD) including the clinical presentation, multimodal ocular imaging, histopathology, and ultrastructural analysis in ten Labrador Retrievers. PROCEDURE: Multicentered data collection. RESULTS: Labrador Retrievers affected by MCD were presented between the age of 4.5 and 6 years of age with a history of cloudy eyes and/or visual impairment. Findings on ophthalmic examination included a diffuse haze of the corneal stroma and multiple, well-demarcated, off-white to yellow-brown, punctate corneal opacities heterogeneous in size. Corneal vascularization developed in most dogs as the disease progressed. Disease progression was associated with increased density of the corneal haze as well as increased number and size of the focal opacities and dogs developed significant visual impairment. Spectral domain-optical coherence tomography revealed multifocal hyper-reflective regions within the stroma. In vivo confocal microscopy revealed marked alterations in reflectivity throughout the entire stroma. Normal keratocytes could not be identified in affected areas. Histopathology showed stromal collagen fibers separated by acidophilic granular material on hematoxylin and eosin stain. The material stained with periodic acid-Schiff and colloidal iron stain but not with Masson trichrome stain, confirming the accumulation of glycosaminoglycans. On electron microscopic ultrastructural examination, keratocytes presented with vacuolated rough endoplasmic reticulum and multiple electron dense cytoplasmic inclusions. In areas keratocytes appeared ruptured, with cell organelles and proteinaceous material grouped together between collagen fibers. CONCLUSION: MCD in Labrador Retrievers has similarities with the human counterpart of the condition and is an important differential diagnosis in dogs with corneal disease.

Identification of Nonconsensual Sexual Experiences and the Sexual Self-Schemas of Women: Implications for Sexual Functioning.

Many individuals who experience nonconsensual sexual experiences (NSEs) do not identify their experiences with common sexual violence labels (e.g., sexual assault, rape, or abuse), and cognitive mechanisms of identification have yet to be examined. Identification may involve the integration of the experience into sexual self-schemas, which would have implications for sexual well-being. Women were recruited through Amazon's Mechanical Turk (N = 818) to take part in an anonymous online study of sexual experiences. The current study assessed the relationship between textually derived sexual self-schemas and sexual function (measured by the Female Sexual Function Index) in women (M = 35.37 years, SD = 11.27) with NSEs who both did (identifiers, n = 305) and did not (non-identifiers, n = 176) identify with common sexual violence labels, in comparison with those with no NSEs (n = 337). Text analyses revealed nine sexual self-schema themes in participants' essays: Virginity, Openness, Erotophilia, NSEs, Romantic, Sexual Activity, Warmth, Relationships, and Reflection. Analyses demonstrated that identifiers reported significantly poorer sexual functioning and less use of both the Warmth and Openness themes than those with no NSEs. Identifiers also invoked the NSE theme more frequently than both those with no NSE histories and non-identifiers. While greater prominence of the Warmth theme was predictive of greater sexual functioning for both non-identifiers and those with no NSEs, this was not true for identifiers. Instead, the NSE theme was significantly predictive of lower sexual functioning in identifiers. The results suggest that NSE identification may result in greater internalization of the NSE into one's sexual self-schema and, in turn, predict decrements in sexual functioning. The results are discussed in relation to identification interpretation and clinical intervention.