Automated extraction and LC-MS-MS analysis of 11-nor-9-carboxy-tetrahydrocannabinol isomers and prevalence in authentic urine specimens.

11-Nor-9-carboxy-Δ9-tetrahydrocannabinol (Δ9-THCCOOH) is the most frequently detected illicit drug metabolite in the military drug testing program. An increasing number of specimens containing unresolved Δ8-THCCOOH prompted the addition of this analyte to the Department of Defense drug testing panel. A method was developed and validated for the quantitative confirmation of the carboxylated metabolites of Δ8- and Δ9-THC in urine samples utilizing automated pipette tip dispersive solid-phase extraction and analysis by liquid chromatography-tandem mass spectrometry (LC-MS-MS). Analytes were separated isocratically over an 8.5-min runtime and detected on an MS-MS equipped with an electrospray ionization source operated in negative mode. A single point calibrator (15 ng/mL) forced through zero demonstrated linearity from 3 to 1,000 ng/mL. Intra- and inter-day precision were ≤9.1%, and bias was within ±14.1% for Δ8-THCCOOH and Δ9-THCCOOH. No interferences were found after challenging the method with different over-the-counter drugs, prescription pharmaceuticals, drugs of abuse and several cannabinoids and cannabinoid metabolites, including Δ10-THCCOOH. Urine specimens presumptively positive by immunoassay (n = 2,939; 50 ng/mL Δ9-THCCOOH cutoff) were confirmed with this analytical method. Δ8-THCCOOH and Δ9-THCCOOH were present together above the 15 ng/mL cutoff in 33% of specimens. However, nearly one-third of the specimens analyzed were positive for Δ8-THCCOOH only. This manuscript describes the first validated automated extraction and confirmation method for Δ8- and Δ9-THCCOOH in urine that provides adequate analyte separation in urine specimens with extreme isomer abundance ratios.

Demographic consequences of an extreme heat wave are mitigated by spatial heterogeneity in an annual monkeyflower.

Heat waves are becoming more frequent and intense with climate change, but the demographic and evolutionary consequences of heat waves are rarely investigated in herbaceous plant species. We examine the consequences of a short but extreme heat wave in Oregon populations of the common yellow monkeyflower (Mimulus guttatus) by leveraging a common garden experiment planted with range-wide populations and observational studies of 11 local populations. In the common garden, 89% of seedlings died during the heat wave including >96% of seedlings from geographically local populations. Some populations from hotter and drier environments had higher fitness, however, others from comparable environments performed poorly. Observational studies of local natural populations drastically differed in the consequences of the heat wave-one population was completely extirpated and nearly half had a >50% decrease in fitness. However, a few populations had greater fitness during the heat wave year. Differences in mortality corresponded to the impact of the heat wave on soil moisture-retention of soil moisture throughout the heat wave led to greater survivorship. Our results suggest that not all populations experience the same intensity or degree of mortality during extreme events and such heterogeneity could be important for genetic rescue or to facilitate the distribution of adaptive variants throughout the region.

Unique drought resistance strategies occur among monkeyflower populations spanning an aridity gradient.

PREMISE: Annual plants often exhibit drought-escape and avoidance strategies to cope with limited water availability. Determining the extent of variation and factors underlying the evolution of divergent strategies is necessary for determining population responses to more frequent and severe droughts. METHODS: We leveraged five Mimulus guttatus populations collected across an aridity gradient within manipulative drought and quantitative genetics experiments to examine constitutive and terminal-drought induced responses in drought resistance traits. RESULTS: Populations varied considerably in drought-escape- and drought-avoidance-associated traits. The most mesic population demonstrated a unique resource conservative strategy. Xeric populations exhibited extreme plasticity when exposed to terminal drought that included flowering earlier at shorter heights, increasing water-use efficiency, and shifting C:N ratios. However, plasticity responses also differed between populations, with two populations slowing growth rates and flowering at earlier nodes and another population increasing growth rate. While nearly all traits were heritable, phenotypic correlations differed substantially between treatments and often, populations. CONCLUSIONS: Our results suggest drought resistance strategies of populations may be finely adapted to local patterns of water availability. Substantial plastic responses suggest that xeric populations can already acclimate to drought through plasticity, but populations not frequently exposed to drought may be more vulnerable.

Motorized 3D Ultrasound and Jugular Vein Dimension Measurement on the International Space Station.

BACKGROUND: Internal jugular vein (IJV) congestion occurs during spaceflight. Historically, IJV distension on the International Space Station (ISS) has been quantified using single slice cross-sectional images from conventional 2D ultrasound with remote guidance. Importantly, the IJV is an irregular shape and highly compressible. Consequently, conventional imaging is susceptible to poor reproducibility due to inconsistent positioning, insonation angle, and hold-down pressure, especially when controlled by novice sonographers (i.e., astronauts). Recently, a motorized 3D ultrasound was launched to the ISS that mitigates angulation errors and has a larger design, allowing for more consistent hold-down pressure and positioning. This short communication compares IJV congestion measured with 2D vs. 3D methods during spaceflight.METHODS: IJV was measured prior to and following a 4-h venoconstrictive thigh cuff countermeasure. Data were acquired from three astronauts approximately halfway through their 6-mo missions.RESULTS: The 2D and 3D ultrasound results were not congruent in all astronauts. 3D ultrasound confirmed that the countermeasure reduced IJV volume in three astronauts by approximately 35%, whereas 2D data were more equivocal. These results indicate that 3D ultrasound provides less error-prone quantitative data.DISCUSSION: These data are the first to compare 2D and 3D methods during spaceflight in the same participants by using a known countermeasure that reduces IJV congestion. The current results demonstrate that 3D ultrasound should be the preferred imaging method when trying to measure venous congestion in the IJV, and that 2D ultrasound results should be interpreted with caution.Patterson C, Greaves DK, Robertson A, Hughson R, Arbeille PL. Motorized 3D ultrasound and jugular vein dimension measurement on the International Space Station. Aerosp Med Hum Perform. 2023; 94(6):466-469.

Assessing jugular venous compliance with optical hemodynamic imaging by modulating intrathoracic pressure.

Significance: The internal jugular veins (IJV) are critical cerebral venous drainage pathways that are affected by right heart function. Cardiovascular disease and microgravity can alter central venous pressure (CVP) and venous return, which may contribute to increased intracranial pressure and decreased cardiac output. Assessing jugular venous compliance may provide insight into cerebral drainage and right heart function, but monitoring changes in vessel volume is challenging. Aim: We investigated the feasibility of quantifying jugular venous compliance from jugular venous attenuation (JVA), a noncontact optical measurement of blood volume, along with CVP from antecubital vein cannulation. Approach: CVP was progressively increased through a guided graded Valsalva maneuver, increasing mouth pressure by 2 mmHg every 2 s until a maximum expiratory pressure of 20 mmHg. JVA was extracted from a 1-cm segment between the clavicle and midneck. The contralateral IJV cross-sectional area (CSA) was measured with ultrasound to validate changes in the vessel size. Compliance was calculated using both JVA and CSA between four-beat averages over the duration of the maneuver. Results: JVA and CSA were strongly correlated (median and interquartile range) over the Valsalva maneuver across participants (r = 0.986, [0.983, 0.987]). CVP more than doubled on average between baseline and peak strain (10.7 ± 4.4 vs. 25.8 ± 5.4 cmH2O; p < 0.01). JVA and CSA increased nonlinearly with CVP, and both JVA- and CSA-derived compliance decreased progressively from baseline to peak strain (49% and 56% median reduction, respectively), with no significant difference in compliance reduction between the two measures (Z = - 1.24, p = 0.21). Pressure-volume curves showed a logarithmic relationship in both CSA and JVA. Conclusions: Optical jugular vein assessment may provide new ways to assess jugular distention and cardiac function.

Implementation of exercise countermeasures during spaceflight and microgravity analogue studies: Developing countermeasure protocols for bedrest in older adults (BROA).

Significant progress has been made in the development of countermeasures to attenuate the negative consequences of prolonged exposure to microgravity on astronauts' bodies. Deconditioning of several organ systems during flight includes losses to cardiorespiratory fitness, muscle mass, bone density and strength. Similar deconditioning also occurs during prolonged bedrest; any protracted time immobile or inactive, especially for unwell older adults (e.g., confined to hospital beds), can lead to similar detrimental health consequences. Due to limitations in physiological research in space, the six-degree head-down tilt bedrest protocol was developed as ground-based analogue to spaceflight. A variety of exercise countermeasures have been tested as interventions to limit detrimental changes and physiological deconditioning of the musculoskeletal and cardiovascular systems. The Canadian Institutes of Health Research and the Canadian Space Agency recently provided funding for research focused on Understanding the Health Impact of Inactivity to study the efficacy of exercise countermeasures in a 14-day randomized clinical trial of six-degree head-down tilt bedrest study in older adults aged 55-65 years old (BROA). Here we will describe the development of a multi-modality countermeasure protocol for the BROA campaign that includes upper- and lower-body resistance exercise and head-down tilt cycle ergometry (high-intensity interval and continuous aerobic exercise training). We provide reasoning for the choice of these modalities following review of the latest available information on exercise as a countermeasure for inactivity and spaceflight-related deconditioning. In summary, this paper sets out to review up-to-date exercise countermeasure research from spaceflight and head-down bedrest studies, whilst providing support for the proposed research countermeasure protocols developed for the bedrest study in older adults.

Sex-dependent jugular vein optical attenuation and distension during head-down tilt and lower body negative pressure.

Non-contact coded hemodynamic imaging (CHI) is a novel wide-field near-infrared spectroscopy system which monitors blood volume by quantifying attenuation of light passing through the underlying vessels. This study tested the hypothesis that CHI-based jugular venous attenuation (JVA) would be larger in men, and change in JVA would be greater in men compared to women during two fluid shift challenges. The association of JVA with ultrasound-based cross-sectional area (CSA) was also tested. Ten men and 10 women completed three levels of head-down tilt (HDT) and four levels of lower body negative pressure (LBNP). Both JVA and CSA were increased by HDT and reduced by LBNP (all p < 0.001). Main effects of sex indicated that JVA was higher in men than women during both HDT (p = 0.003) and LBNP (p = 0.011). Interaction effects of sex and condition were observed for JVA during HDT (p = 0.005) and LBNP (p < 0.001). We observed moderate repeated-measures correlations (rrm ) between JVA and CSA in women during HDT (rrm  = 0.57, p = 0.011) and in both men (rrm  = 0.74, p < 0.001) and women (rrm  = 0.66, p < 0.001) during LBNP. While median within-person correlation coefficients indicated an even stronger association between JVA and CSA, this association became unreliable for small changes in CSA. As hypothesized, JVA was greater and changed more in men compared to women during both HDT and LBNP. CHI provides a non-contact method of tracking large changes in internal jugular vein blood volume that occur with acute fluid shifts, but data should be interpreted in a sex-dependent manner.

Suicidal Ideation, Planning, and Attempts Among Synthetic Cannabinoid Users Across Different Demographic Subgroups.

Background: Case studies have linked synthetic cannabinoid (SC) use to suicide risk both during and following acute psychoactive effects. No study has explored whether a meaningful relationship exists between SC use and suicidal ideation, planning, and attempts within a generalizable sample. Aims: This work attempted to determine whether SC use is significantly associated with suicidal thoughts, planning, or attempts and whether the association between SC use and suicide may be stronger in certain demographic groups. Method: Cross-sectional self-report data from the Youth Risk Behavior Surveillance System were utilized. Logistic regression models were explored within a sample of 29,384 adolescents. Results: A clear connection emerged between SC use and suicidal thoughts and attempts. SC use and suicide risk were strongly related within the lesbian, gay, and bisexual and African Americans populations, even after controlling for depressive symptoms. Limitations: The study was cross-sectional and respondents may have underreported substance use or suicidal thoughts. Conclusion: Whether SC use has an impact on suicidal ideation directly, indirectly, or if it is simply a co-occurring phenomenon, it may serve as a flag of enhanced risk. Physicians must remain cognizant of the SC-suicide risk connection, particularly among African American and/or lesbian, gay, or bisexual patients.

Inhibition of DNA amplification caused by metal in extracted bloodstains and in direct amplification.

Copper is a metal that is commonly found in many evidence types, such as weapons, and household items, that are often retrieved from crime scenes. We used copper and brass sheets, cartridge cases, and metal décor items as substrates for deposition of blood to study inhibition induced by metals. Another objective was to compare the efficiency of STR profiling following extraction, quantification, amplification, and capillary electrophoresis with a direct amplification method which only requires amplification and capillary electrophoresis. Samples that were designated for extraction, were collected, extracted, and purified using the Qiagen EZ1 DNA Investigator® Kit. Direct amplification was performed using Copan microFLOQ® Direct Swabs to collect minute amount of blood. Complete STR profiles were obtained from 54% of the extracted samples and from 88% of directly amplified samples. Substrate type and amplification method (two-way ANOVA, F(5, 131) = 3.59, p = 4.50 × 10-3) impacted profile completeness but collection time did not (one-way ANOVA, F(1, 131) = 0.017, p = 0.90). Composition of the substrates was determined with scanning electron microscopy (SEM), and inhibition caused by copper was evaluated for each substrate. When substrate composition was analyzed in combination with amplification method, a substrate's copper percent was not associated with increased allelic dropout (one-way ANOVA, F(1, 176) = 0.70, p = 0.40) but amplification alone was statistically significant (one-way ANOVA, F(1, 176) = 40.64, p = 1.56 × 10-9). Using the Copan microFLOQ® Direct Swabs, inhibition induced by metal observed was not as pronounced. In addition, less variability in profile completeness was detected, profiles were generated within a very short period, and only minute amount of bloodstain was needed, allowing for additional analysis of the same samples. Our results directly quantify the effect of copper composition on DNA profiling and suggest that while copper percentage is an observational contributor to allele dropout it is not wholly responsible for these events.

Optical Hemodynamic Imaging of Jugular Venous Dynamics During Altered Central Venous Pressure.

OBJECTIVE: An optical imaging system is proposed for quantitatively assessing jugular venous response to altered central venous pressure. METHODS: The proposed system assesses sub-surface optical absorption changes from jugular venous waveforms with a spatial calibration procedure to normalize incident tissue illumination. Widefield frames of the right lateral neck were captured and calibrated using a novel flexible surface calibration method. A hemodynamic optical model was derived to quantify jugular venous optical attenuation (JVA) signals, and generate a spatial jugular venous pulsatility map. JVA was assessed in three cardiovascular protocols that altered central venous pressure: acute central hypovolemia (lower body negative pressure), venous congestion (head-down tilt), and impaired cardiac filling (Valsalva maneuver). RESULTS: JVA waveforms exhibited biphasic wave properties consistent with jugular venous pulse dynamics when time-aligned with an electrocardiogram. JVA correlated strongly (median, interquartile range) with invasive central venous pressure during graded central hypovolemia (r = 0.85, [0.72, 0.95]), graded venous congestion (r = 0.94, [0.84, 0.99]), and impaired cardiac filling (r = 0.94, [0.85, 0.99]). Reduced JVA during graded acute hypovolemia was strongly correlated with reductions in stroke volume (SV) (r = 0.85, [0.76, 0.92]) from baseline (SV: 79 ± 15 mL, JVA: 0.56 ± 0.10 a.u.) to -40 mmHg suction (SV: 59 ± 18 mL, JVA: 0.47 ± 0.05 a.u.; p 0.01). CONCLUSION: The proposed non-contact optical imaging system demonstrated jugular venous dynamics consistent with invasive central venous monitoring during three protocols that altered central venous pressure. SIGNIFICANCE: This system provides non-invasive monitoring of pressure-induced jugular venous dynamics in clinically relevant conditions where catheterization is traditionally required, enabling monitoring in non-surgical environments.

How discourse constraints influence neurolinguistic mechanisms during the comprehension of proverbs.

We use event-related brain potential (ERP) methodology to examine the influence of the linguistic markers literally speaking and figuratively speaking on the comprehension of proverbs (e.g., The cat is out of the bag). Our results show that slow cortical potentials at anterior electrode sites varied in amplitude across the proverbs as a function of the presence or absence of the markers, the presence and absence of discourse contexts, and the familiarity of the proverbs. The results demonstrate that the integration of literal meaning into context is easier than figurative meaning, and argues against models of figurative language processing that hold that comprehenders are obligated either to first process the literal or figurative sense of the trope.

Co-occurring nonnative woody shrubs have additive and non-additive soil legacies.

To maximize limited conservation funds and prioritize management projects that are likely to succeed, accurate assessment of invasive nonnative species impacts is essential. A common challenge to prioritization is a limited knowledge of the difference between the impacts of a single nonnative species compared to the impacts of nonnative species when they co-occur, and in particular predicting when impacts of co-occurring nonnative species will be non-additive. Understanding non-additivity is important for management decisions because the management of only one co-occurring invader will not necessarily lead to a predictable reduction in the impact or growth of the other nonnative plant. Nonnative plants are frequently associated with changes in soil biotic and abiotic characteristics, which lead to plant-soil interactions that influence the performance of other species grown in those soils. Whether co-occurring nonnative plants alter soil properties additively or non-additively relative to their effects on soils when they grow in monoculture is rarely addressed. We use a greenhouse plant-soil feedback experiment to test for non-additive soil impacts of two common invasive nonnative woody shrubs, Lonicera maackii and Ligustrum sinense, in deciduous forests of the southeastern United States. We measured the performance of each nonnative shrub, a native herbaceous community, and a nonnative woody vine in soils conditioned by each shrub singly or together in polyculture. Soils conditioned by both nonnative shrubs had non-additive impacts on native and nonnative performance. Root mass of the native herbaceous community was 1.5 times lower and the root mass of the nonnative L. sinense was 1.8 times higher in soils conditioned by both L. maackii and L. sinense than expected based upon growth in soils conditioned by either shrub singly. This result indicates that when these two nonnative shrubs co-occur, their influence on soils disproportionally favors persistence of the nonnative L. sinense relative to this native herbaceous community, and could provide an explanation of why native species abundance is frequently depressed in these communities. Additionally, the difference between native and nonnative performance demonstrates that invasive impact studies focusing on the impact only of single species can be insufficient for determining the impact of co-occurring invasive plant species.

Ant-mediated seed dispersal in a warmed world.

Climate change affects communities both directly and indirectly via changes in interspecific interactions. One such interaction that may be altered under climate change is the ant-plant seed dispersal mutualism common in deciduous forests of eastern North America. As climatic warming alters the abundance and activity levels of ants, the potential exists for shifts in rates of ant-mediated seed dispersal. We used an experimental temperature manipulation at two sites in the eastern US (Harvard Forest in Massachusetts and Duke Forest in North Carolina) to examine the potential impacts of climatic warming on overall rates of seed dispersal (using Asarum canadense seeds) as well as species-specific rates of seed dispersal at the Duke Forest site. We also examined the relationship between ant critical thermal maxima (CTmax) and the mean seed removal temperature for each ant species. We found that seed removal rates did not change as a result of experimental warming at either study site, nor were there any changes in species-specific rates of seed dispersal. There was, however, a positive relationship between CTmax and mean seed removal temperature, whereby species with higher CTmax removed more seeds at hotter temperatures. The temperature at which seeds were removed was influenced by experimental warming as well as diurnal and day-to-day fluctuations in temperature. Taken together, our results suggest that while temperature may play a role in regulating seed removal by ants, ant plant seed-dispersal mutualisms may be more robust to climate change than currently assumed.

Best practices in specialty pharmacy management.

BACKGROUND: Specialty pharmacy is a growing area of research, utilization, and cost. Because of the unique nature of the diseases treated by specialty pharmaceuticals, such as cancer and rheumatoid arthritis, novel management approaches are needed. Advocate Physician Partners (APP) is an entity within the Advocate Health Care Health System in the Chicago and the central Illinois area. It coordinates the care management and managed care contracting between the Advocate Health Care System and more than 4,000 physicians on the medical staffs of Advocate hospitals. APP has experienced a per-member-per-month (PMPM) increase of less than 3% in oncology intravenous medications spend in 2012. This spend refers to the intravenous medications covered under the medical benefits for APP's health maintenance organization (HMO) population. The spend has consistently been less than national projections, and we believe this is tied to the adoption of several key best practices. Prior to instituting the best practices, the yearly percentage increases for oncology spending were 5.52% (2007 to 2008), 9.39% (2008 to 2009), and 5.29% (2009 to 2010). After instituting best practices during the first quarter of 2011, the increases in PMPM were 3.11% (2010 to 2011) and 2.11% (2011 to 2012), which were below previous years. OBJECTIVE: To describe the best practices of specialty pharmacy management adopted by APP, specifically (a) establishing a content expert and governing bodies, (b) ensuring compliance with policies, and (c) providing educational resources. METHODS: APP has several key result areas (KRAs). One KRA was compliance with appropriate utilization of intravenous oncology protocols for its HMO population. The protocols for each medication outline the appropriate indication and patient population. These protocols were developed and reviewed by the APP Pharmacy and Technology (PT) committee. The PT-approved indications reflect FDA indications and indications found in national guidelines. The APP KRA target for the utilization of protocols was 80%. The compliance for completing the protocols that correspond to these medications was calculated by tabulating the number of paid claims over the number of completed protocols, resulting in a compliance percentage. APP defined noncompliance as any utilization not outlined in the protocol. Another KRA was physician feedback. APP requires that physicians complete a certain number of continuing medical education (CME) programs provided by APP each year. Feedback from physicians, provided at the end of the CME, were tabulated and utilized for further programs. Additionally, APP strived to increase physician engagement by placing them in key roles that oversaw clinical and business aspects of the organization. In order to meet the KRAs, investigate inappropriate utilization, and become better involved in engaging oncology specialists, APP invested in an oncology clinical pharmacist position. The primary responsibility of the oncology clinical pharmacist was to be a specialty drug resource for the organization with a focus on provider education. The oncology clinical pharmacist was deemed the content expert who developed clinical protocols, educated providers, and encouraged compliance with organizational policies and procedures as it pertained to the KRAs. RESULTS: Since establishing the oncology clinical pharmacist position, APP has seen an increase in protocol compliance. Prior to the institution of this position, the protocol compliance was 62%. In less than 1 year after hiring the oncology clinical pharmacist, the compliance percentage spiked to over 80%. APP has several committees and boards that oversee the clinical and business aspects of the organization. By placing physicians in chairmanship positions of the committees, APP has seen the benefit of handling difficult patient decisions and increased compliance with policies. Lastly, extensive provider education has led to an improved physician satisfaction in the educational initiatives of APP. Greater than 80% of the physicians felt the CME content was relevant to their practices; the content would likely have a positive impact on their practices; and the clinical content was evidence based and accepted by the medical community. CONCLUSIONS: By hiring an oncology clinical pharmacist, engaging physicians by placing them in key roles, and providing more specialist-specific education, APP has improved its KRAs and compliance percentages. APP achieved success in containing expenditures for oncology intravenous medications by implementing key best practices combined with traditional management strategies.

Electrophysiological evidence for the time-course of verifying text ideas.

We examined how verb factivity influences the ability of readers to detect and resolve the mismatch of receiving false referents in relation to true referents in discourse contexts. Factive verbs (e.g., know), but not nonfactive verbs (believe), entail the truth of their complements. Recent research by Singer [Singer, M. (2006). Verification of text ideas during reading. Journal of Memory and Language, 54, 574-591] suggests that there are pragmatic costs associated with knowing something that is clearly false and only believing something that is clearly true. However, because Singer measured reading times for full sentences, it could not be determined whether these costs were initiated upon the appearance of the critical target word (i.e., the word that validated or invalidated previous text ideas) or at a later point in the sentences. In the present research we recorded event-related brain potentials while people read the same passages for comprehension and analyzed potentials evoked to the critical target words. Our results demonstrate that the brain distinguishes between true and false target words by at least 200ms after their onset, and that the pragmatic costs identified by Singer lead to interactions between verb factivity and truth in both early (P2) and later occurring brain components (late phase of N400 and late frontal positivity). In general, the results suggest readers had greater difficulty integrating false nouns than true nouns following factive than nonfactive verbs, and that detection of this mismatch also occurred earlier following factive verbs. Our results provide insight into the time-course of the processes that underlie the verification of text ideas, and extend neurocognitive research on anaphoric resolution.