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Ascites reprograms innate lymphoid immune cells in ovarian cancer by promoting ILC2 enrichment and dysfunctional NK-cell states.

BACKGROUND: High-grade serous ovarian cancer (HGSOC) is commonly accompanied by malignant ascites, a clinically relevant tumor niche that promotes immune evasion, metastasis, and treatment resistance. Although natural killer (NK)-cell dysfunction has been described in ovarian cancer, the broader innate lymphoid landscape of ascites and the mechanisms linking ascites-derived signals to innate immune suppression remain insufficiently resolved. METHODS: We performed single-cell RNA sequencing of NK/innate lymphoid cells from ovarian cancer ascites to define cellular heterogeneity and differentiation states. Functional assays assessed NK-cell cytotoxicity, degranulation, and receptor expression following exposure to patient-derived ascites, with or without transforming growth factor-β (TGF-β) receptor inhibition. Proteomic profiling was used to characterize the soluble ascites milieu, and clinical associations were examined for innate lymphoid subsets. RESULTS: Single-cell analysis identified eight transcriptionally distinct NK/innate lymphoid states, including cytotoxic, precursor, early-like, tolerant/immunoregulatory, regulatory, proinflammatory, and innate lymphoid populations. Ovarian cancer ascites was characterized by depletion of cytotoxic and precursor NK-cell states together with enrichment of early-like, tolerant, regulatory, pro-inflammatory, and innate lymphoid cell (ILC) populations. Trajectory analysis indicated impaired maturation toward terminally differentiated cytotoxic NK cells. Notably, ascites contained an expanded population of programmed cell death protein 1 (PD-1)+ ILC2s, which were more abundant in patients with shorter progression-free survival. In functional assays, short-term exposure of healthy donor NK cells to ascites suppressed degranulation and tumor-cell killing, reduced expression of activating receptors including NKp30 and DNAM-1, increased inhibitory receptor expression, and shifted NK cells toward a CD56highCD16low phenotype. Proteomic profiling supported a soluble milieu consistent with type 2 immune skewing and NK-cell suppression. Importantly, TGF-β receptor inhibition partially restored NK-cell activation and function in the presence of ascites. CONCLUSIONS: HGSOC ascites establishes a type 2-skewed immunoregulatory niche that coordinately drives NK cell dysfunction and PD-1+ ILC2 accumulation. The findings identify TGF-β-linked suppression and ascites-associated immune regulators as candidate immunotherapeutic vulnerabilities for restoring antitumor immunity in ovarian cancer.

Humans↗

Analysis for chemical characterization of atmospheric aerosols application of X-ray microprobe system and double thin film method.

The X-ray microprobe system was applied to ultra trace characterization of single Kosa aerosols and non-Kosa aerosols simultaneously collected at Yasaka, Japan and TaeAnn, Korea. We demonstrated remarkable mass increase of heavy metals as well as soil components in individual particles during the Kosa event compared with the non-Kosa period at Yasaka. Backward trajectory analysis suggested that the Kosa samples were in the mixing state of mineral components and anthropogenic heavy elements. Double thin film method was applied to investigate the seasonal change of the mixing states of single sea-salt aerosols associated with chlorine loss due to the heterogeneous reactions between sea-salt particles and acidic gases. It was revealed that the percentages of both chloride-nitrate mixed particles and sulfate-nitrate ones were larger in wintertime than those in summertime and fraction of chloride-nitrate mixed particles increased with an increase of particle size. Comparison between the size-segregated bulk analysis and the single particle analysis demonstrated that remarkable chloride depletion occurred in coarse particles sampled on May in the former analysis, while chloride depletion in coarse particles was not marked in the latter analysis. The discrepancy attributes to the difference of the sampling time between both analyses because significant change of air mass route occurred during the bulk sampling after completion of the single particle sampling.

Aerosols↗

Automatic tracking of individual fluorescence particles: application to the study of chromosome dynamics.

We present a new, robust, computational procedure for tracking fluorescent markers in time-lapse microscopy. The algorithm is optimized for finding the time-trajectory of single particles in very noisy dynamic (two- or three-dimensional) image sequences. It proceeds in three steps. First, the images are aligned to compensate for the movement of the biological structure under investigation. Second, the particle's signature is enhanced by applying a Mexican hat filter, which we show to be the optimal detector of a Gaussian-like spot in 1/omega2 noise. Finally, the optimal trajectory of the particle is extracted by applying a dynamic programming optimization procedure. We have used this software, which is implemented as a Java plug-in for the public-domain ImageJ software, to track the movement of chromosomal loci within nuclei of budding yeast cells. Besides reducing trajectory analysis time by several 100-fold, we achieve high reproducibility and accuracy of tracking. The application of the method to yeast chromatin dynamics reveals different classes of constraints on mobility of telomeres, reflecting differences in nuclear envelope association. The generic nature of the software allows application to a variety of similar biological imaging tasks that require the extraction and quantitation of a moving particle's trajectory.

Algorithms↗

Motor task difficulty and brain activity: investigation of goal-directed reciprocal aiming using positron emission tomography.

Differences in the kinematics and pattern of relative regional cerebral blood flow (rCBF) during goal-directed arm aiming were investigated with the use of a Fitts continuous aiming paradigm with three difficulty conditions (index of difficulty, ID) and two aiming types (transport vs. targeting) in six healthy right-handed young participants with the use of video-based movement trajectory analysis and positron emission tomography. Movement time and kinematic characteristics were analyzed together with the magnitude of cerebral blood flow to identify areas of brain activity proportionate to task and movement variables. Significant differences in rCBF between task conditions were determined by analysis of variance with planned comparisons of means with the use of group mean weighted linear contrasts. Data were first analyzed for the group. Then individual subject differences for the movement versus no movement and task difficulty comparisons were related to each individual subjects' anatomy by magnetic resonance imaging. Significant differences in rCBF during reciprocal aiming compared with no-movement conditions were found in a mosaic of well-known cortical and subcortical areas associated with the planning and execution of goal-directed movements. These included cortical areas in the left sensorimotor, dorsal premotor, and ventral premotor cortices, caudal supplementary motor area (SMA) proper, and parietal cortex, and subcortical areas in the left putamen, globus pallidus, red nucleus, thalamus, and anterior cerebellum. As aiming task difficulty (ID) increased, rCBF increased in areas associated with the planning of more complex movements requiring greater visuomotor processing. These included bilateral occipital, left inferior parietal, and left dorsal cingulate cortices--caudal SMA proper and right dorsal premotor area. These same areas showed significant increases or decreases, respectively, when contrast means were compared with the use of movement time or relative acceleration time, respectively, as the weighting factor. Analysis of individual subject differences revealed a correspondence between the spatial extent of rCBF changes as a function of task ID and the individuals' movement times. As task ID decreased, significant increases in rCBF were evident in the right anterior cerebellum, left middle occipital gyrus, and right ventral premotor area. Functionally, these areas are associated with aiming conditions in which the motor execution demands are high (i.e., coordination of rapid reversals) and precise trajectory planning is minimal. These same areas showed significant increases or decreases, respectively, when contrast means were compared with the use of movement time or relative acceleration time, respectively, as the weighting factor. A functional dissociation resulted from the weighted linear contrasts between larger (limb transport) or smaller (endpoint targeting) type amplitude/target width aiming conditions. Areas with significantly greater rCBF for targeting were the left motor cortex, left intraparietal sulcus, and left caudate. In contrast, those areas with greater rCBF associated with limb transport included bilateral occipital lingual gyri and the right anterior cerebellum. Various theoretical explanations for the speed/accuracy tradeoffs of rapid aiming movements have been proposed since the original information theory hypothesis of Fitts. This is the first report to relate the predictable variations in motor control under changing task constraints with the functional anatomy of these rapid goal-directed aiming movements. Differences in unimanual aiming task difficulty lead to dissociable activation of cortical-subcortical networks. Further, these data suggest that when more precise targeting is required, independent of task difficulty, a cortical-subcortical loop composed of the contralateral motor cortex, intraparietal sulcus, and caudate is activated. This is consistent with the role of motor cortex

Adult↗

Molecular simulation study of structural and dynamic properties of mixed DPPC/DPPE bilayers.

Molecular dynamics simulations have been used to study structural and dynamic properties of fully hydrated mixed 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE) bilayers at 0, 25, 50, 75, and 100 mol % DPPE. Simulations were performed for 50 ns at 350 K and 1 bar for the liquid-crystalline state of the mixtures. Results show that the average area per headgroup reduces from 0.65 +/- 0.01 nm(2) in pure DPPC to 0.52 +/- 0.01 nm(2) in pure DPPE systems. The lipid tails become more ordered with increasing DPPE concentration, resulting in a slight increase in membrane thickness (3.43 +/- 0.01 nm in pure DPPC to 4.00 +/- 0.01 nm in pure DPPE). The calculated area per headgroup and order parameter for pure DPPE deviates significantly from available experimental measurements, suggesting that the force field employed requires further refinement. In-depth analysis of the hydrogen-bond distribution in DPPE molecules shows that the amine groups strongly interact with the phosphate and carbonyl groups through inter/intramolecular hydrogen bonds. This yields a bilayer structure with DPPE headgroups preferentially located near the lipid phosphate and ester oxygens. It is observed that increasing DPPE concentrations causes competitive hydrogen bonding between the amine groups (hydrogen-donor) and the phosphate/carbonyl groups or water (hydrogen-acceptor). Due to the increasing number of hydrogen-donors from DPPE molecules with increasing concentration, DPPE becomes more hydrated. Trajectory analysis shows that DPPE molecules in the lipid mixtures move laterally and randomly around the membrane surface and the movement becomes more localized with increasing DPPE concentrations. For the conditions and simulation time considered, no aggregation or phase separation was observed between DPPC and DPPE.

1,2-Dipalmitoylphosphatidylcholine↗

Measurement and analysis of the relationship between ammonia, acid gases, and fine particles in eastern North Carolina.

An annular denuder system, which consisted of a cyclone separator; two diffusion denuders coated with sodium carbonate and citric acid, respectively; and a filter pack consisting of Teflon and nylon filters in series, was used to measure acid gases, ammonia (NH3), and fine particles in the atmosphere from April 1998 to March 1999 in eastern North Carolina (i.e., an NH3-rich environment). The sodium carbonate denuders yielded average acid gas concentrations of 0.23 microg/m3 hydrochloric acid (standard deviation [SD] +/- 0.2 microg/m3); 1.14 microg/m3 nitric acid (SD +/- 0.81 microg/m3), and 1.61 microg/m3 sulfuric acid (SD +/- 1.58 microg/m3). The citric acid denuders yielded an average concentration of 17.89 microg/m3 NH3 (SD +/- 15.03 microg/m3). The filters yielded average fine aerosol concentrations of 1.64 microg/m3 ammonium (NH4+; SD +/- 1.26 microg/m3); 0.26 microg/m3 chloride (SD +/- 0.69 microg/m3), 1.92 microg/m3 nitrate (SD +/- 1.09 microg/m3), and 3.18 microg/m3 sulfate (SO4(2-); SD +/- 3.12 microg/m3). From seasonal variation, the measured particulates (NH4+, SO4(2-), and nitrate) showed larger peak concentrations during summer, suggesting that the gas-to-particle conversion was efficient during summer. The aerosol fraction in this study area indicated the domination of ammonium sulfate particles because of the local abundance of NH3, and the long-range transport of SO4(2-) based on back trajectory analysis. Relative humidity effects on gas-to-particle conversion processes were analyzed by particulate NH4+ concentration originally formed from the neutralization processes with the secondary pollutants in the atmosphere.

Acids↗

The characteristics of tropospheric ozone seasonality observed from ozone soundings at Pohang, Korea.

This paper presents the first analysis of vertical ozone sounding measurements over Pohang, Korea. The main focus is to analyze the seasonal variation of vertical ozone profiles and determine the mechanisms controlling ozone seasonality. The maxima ozone at the surface and in the free troposphere are observed in May and June, respectively. In comparison with the ozone seasonality at Oki (near sea level) and Happo (altitude of 1840 m) in Japan, which are located at the same latitude as of Pohang, we have found that the time of the ozone maximum at the Japanese sites is always a month earlier than at Pohang. Analysis of the wind flow at the surface shows that the wind shifts from westerly to southerly in May over Japan, but in June over Pohang. However, this wind shift above boundary layer occurs a month later. This wind shift results in significantly smaller amounts of ozone because the southerly wind brings clean wet tropical air. It has been suggested that the spring ozone maximum in the lower troposphere is due to polluted air transported from China. However, an enhanced ozone amount over the free troposphere in June appears to have a different origin. A tongue-like structure in the time-height cross-section of ozone concentrations, which starts from the stratosphere and extends to the middle troposphere, suggests that the ozone enhancement occurs due to a gradual migration of ozone from the stratosphere. The high frequency of dry air with elevated ozone concentrations in the upper troposphere in June suggests that the air is transported from the stratosphere. HYSPLIT trajectory analysis supports the hypothesis that enhanced ozone in the free troposphere is not likely due to transport from sources of anthropogenic activity.

Atmosphere↗

Analysis of dynamics trajectories of DNA and DNA-drug complexes.

A FORTRAN-77 program is described which was applied for analysis of optimized structures and computer-generated dynamics trajectories of DNA and DNA-drug complexes. The CORDAN program (coordinates analysis) also can be used for various manipulations of DNA-drug complexes, i.e. inversion of asymmetric sites or rebuilding the structure of the intercalator, among others. These procedures can find application in drug design. Analysis of dynamics trajectory of neocarzinostatin antibiotic (NCS) intercalated to the A-DNA form of 5'GGATGGGAG:5'CTCCCATCC is presented. The procedures described can be used for detailed analysis of dynamics structures of DNA and their complexes with intercalating drugs.

Base Sequence↗

Molecular dynamics of sickle and normal hemoglobins.

Molecular dynamics (MD) simulations have been carried out for 62.5 ps on crystal structures of deoxy sickle cell hemoglobin (HbS) and normal deoxy hemoglobin (HbA) using the CHARMM MD algorithm, with a time step of 0.001 ps. In the trajectory analysis of the 12.5-62.5 (50 ps) simulation, oscillations of the radius of gyration and solvent-accessible surface area were calculated. HbS exhibited a general contraction during the simulation, while HbA exhibited a nearly constant size. The average deviations of simulated structures from the starting structures were found to be 1.8 A for HbA and 2.3 A for HbS. The average rms amplitudes of atomic motions (atomic flexibility) were about 0.7 A HbA and about 1.0 A for HbS. The amplitudes of backbone motion correlate well with temperature factors derived from x-ray crystallography. A comparison of flexibility between the alpha- and beta-chains in both HbA and HbS indicates that the beta-chains generally exhibited greater flexibility than the alpha-chains, and that the HbS beta-chains exhibit greater flexibility in the N-terminal and D- and F-helix regions than do those of HbA. The average amplitude of backbone torsional oscillations was about 9 degrees, a value comparable with that of other simulations, with enhanced torsional oscillation occurring primarily at the ends of helices or in loop regions between helices. Comparison of atomic flexibility and torsional oscillation results suggests that the increased beta-chain flexibility results from relatively concerted motions of secondary structure elements. The increased flexibility may play an important role in HbS polymerization.(ABSTRACT TRUNCATED AT 250 WORDS)

Computer Simulation↗

Rapid in vivo labeling of identified zebrafish neurons.

We report a simple and rapid method to label individual neurons in live zebrafish embryos and to examine their gene expression profiles. Injection of plasmid DNA encoding an alpha-tubulin promotor driving GFP expression results in mosaic embryos containing a limited number of GFP-positive neurons. Labeled neurons express GFP in their soma and axon, providing the opportunity to analyze pathfinding behaviors of identified neurons in vivo. Moreover, the presence of only a small subset of GFP tagged neurons permits the rapid anatomical identification of these neurons based on soma position and axonal trajectory. Analysis of injected embryos reveals that most, if not all, spinal cord cell types and many other neuronal cell types elsewhere in the nervous system can be GFP tagged. Finally, by combining GFP labeling of individual neurons with fluorescent in situ hybridization, we demonstrate the potential of this method to elucidate gene expression patterns at single cell resolution.

Animals↗

The Effect of Slight Deformation on Thermocapillary-Driven Droplet Coalescence and Growth.

The collision efficiency of two slightly deformable drops in thermocapillary motion at small Reynolds and Marangoni numbers is determined by a trajectory analysis involving methodology from matched asymptotic expansions. The outer solution for two spherical drops which are nearly touching provides the contact force driving the inner solution. Accurate calculation of the contact force and near-contact motion is aided by new solutions for the mobility functions parallel and normal to the drops' line of centers that are valid at very small separations. Governed by a system of integro-differential equations coupling the flow inside the drops and that within the small gap, the inner solution allows demarcation of the regions of drop coalescence and separation. Apart from the driving force, the thin-film equations are unchanged to leading order from the buoyancy-driven case, since no additional singularity is introduced into the tangential stress by the presence of the finite temperature gradient. The interplay of small deformation, as measured by the capillary number (Ca), and attractive van der Waals forces controls the apparent contact motion. Results for the collision efficiency are mapped out for a range of five dimensionless parameters: Ca, size ratio, drop-to-medium viscosity ratio, drop-to-medium thermal conductivity ratio (&kcirc;), and a dimensionless Hamaker parameter. Since the only effect on the inner solution of an increase in the thermal conductivity ratio is an increase in the amount of time the drops spend in close approach, it is possible for the collision efficiency of two slightly deformable drops with higher &kcirc; to be greater than that for two similar drops with lower &kcirc;. This behavior differs from that of spherical drops, where an increase in thermal conductivity ratio always leads to a decrease in the collision efficiency, as a result of greater hydrodynamic interaction between the spherical drops due to the temperature gradient. In addition, collision efficiencies are provided for a model system of ethyl salicylate (ES) drops in diethylene glycol (DEG). The collision efficiency decreases rapidly with increasing drop size above a critical value, due to the increasing role of deformation in retarding the drainage of the thin film between two drops in close approach. Population dynamics simulations are performed for homogeneous suspensions of the ES/DEG system, showing that slight deformation limits droplet growth due to coalescence in dilute dispersions. Copyright 1999 Academic Press.

Journal Article↗

Experimental Investigation of the Orthokinetic Coalescence Efficiency of Droplets in Simple Shear Flow.

The coalescence efficiency of two droplets in simple shear flow was experimentally investigated. The effects of the radius of the colliding droplets, the viscosity of the continuous medium, and the radii ratio on the coalescence efficiency were studied. The coalescence efficiency was determined from the critical angle, alpha(crit), above which the droplets coalesce and below which no coalescence takes place. It was found that the coalescence efficiency decreases and then increases with the flow number. The theories of the drainage models and the trajectory analysis were unable to predict the experimental results. This was attributed to the inadequacy of the existing expression used for the critical film thickness. On the basis of the experimental data obtained a new expression for the critical film thickness was proposed. The new expression predicts a decrease followed by an increase in the critical film thickness. The increase in the critical film thickness is, perhaps, due to instabilities that take place at the interfaces. Implementing the proposed expression for the critical film thickness, the experimentally measured coalescence efficiency, the capture angle and the contact time were successfully predicted. The experimental investigation revealed that the coalescence efficiency does not depend on the radii ratio in the range 0.7 to 1. The experimental investigation also showed that the coalescence efficiency increases as the viscosity of the continuous phase decreases. Measuring the capture angle showed that coalescence can take place either in the compression or in the extensional quadrant of the shear plane. A variation in the capture angle and the contact time was observed for a given fixed set of conditions. The change of the angle alpha with time was measured. The measured values were compared to those calculated from an equation derived by R. S. Allan and S. G. Mason (J. Colloid Interface Sci. 17, pp. 383, 1962). A good agreement was obtained when the droplets were of equal radii and small in size. When the droplets were large in size and of different radii, the agreement was not satisfactory. Measuring the distance between the centers of the colliding droplets showed that the two droplets vibrated when they came close to each other. Copyright 2001 Academic Press.

Journal Article↗

The change in O3, SO2 and NO2 concentrations in Lithuania.

Due to the dynamic nature of the atmosphere, substantial amounts of gaseous and particulate pollutants are transported to the areas distant from their sources. In order to determine the regional concentration levels of atmospheric pollutants in Lithuania, concentrations of gaseous O3, SO2, NO2 and other pollutants have been measured at the Preila background station (55 degrees 20' N and 21 degrees 00' E, 5 m a.s.l.) since 1981. The long-term concentration data set enabled us to get temporal trends, both on a seasonal and longer time scale, to identify source areas of pollutants and to relate them to the emission data. Based on the data obtained, the different tendencies in the pollutant concentration changes were revealed. Positive trends for ozone (of 2.9% per year during 1983-2000) and a distinct negative trend for both sulphur dioxide (of 3.8% per year during 1981-2000) and nitrogen dioxide (of 3.8% per year during 1983-2000) were found. The air mass back-trajectory analysis was used to assess the source region of air pollutants transported to Lithuania. The pollutant concentration levels were compared with their emission changes in Europe and Lithuania. The general trends in SO2 as well as in NO2 concentrations observed are consistent with changes in SO2 and NO2 emissions in Europe and Lithuania.

Air Movements↗

Effectiveness of a community-based advocacy and learning program for hmong refugees.

The effectiveness of a community-based advocacy and learning intervention for Hmong refugees was assessed using a comprehensive, multi-method strategy, which included a within-group longitudinal design with four data collection points and in-depth qualitative recruitment and post-intervention interviews. The intervention's impact on five aspects of refugee well-being was examined: Participants' psychological well-being, quality of life, access to resources, English proficiency, and knowledge for the U.S. citizenship exam. Twenty-eight Hmong adults and 27 undergraduate students participated together in the intervention, which had two major components: (1) Learning Circles, which involved cultural exchange and one-on-one learning opportunities for Hmong adults, and (2) an advocacy component that involved undergraduates advocating for and transferring advocacy skills to Hmong families to increase their access to resources in their communities. Undergraduate paraprofessionals and Hmong participants worked together for 6-8 hr per week for 6 months. Growth trajectory analysis revealed promising quantitative findings. Participants' quality of life, satisfaction with resources, English proficiency, and knowledge for the U.S. citizenship test increased and their levels of distress decreased over the course of the intervention. Mediating analyses suggested that participants' increased quality of life could be explained by their improved satisfaction with resources. Qualitative data helped to support and explain the quantitative data, as well as providing insight into other outcomes and processes of the intervention. Policy, practice, and research implications are discussed.

Adolescent↗

Evaluation of carbon uptake and emissions by forests in Korea during the last thirty years (1973-2002).

The contribution of Korean forests to carbon sequestration for anthropogenic carbon emissions was evaluated. In addition, monitoring of carbon species released from forest fires was conducted. Despite a high carbon uptake by Korean forests, a tremendous increase in fossil fuel burning resulted in a small contribution by forests to carbon removal. The removal efficiency had a 5-31% range with an average of 12% during the period 1973-2002. In 2000, the amount of carbon released from burned trees corresponded to 1.6% of carbon uptake by forests. The distribution of surface CO concentration (ppb) derived from MOPITT (Measurement of Pollution in the Troposphere) showed high CO levels over the East/Japan Sea on April 10, 2000 when the largest forest fires occurred along the east coast of Korea. Trajectory analysis and ground CO measurements also indicated that CO levels over the East/Japan Sea were influenced by forest fires. This study suggests that continuous monitoring of carbon emissions from forest fires is needed for a more reliable estimate of carbon flux in the environment.

Carbon↗

Hemispheric control of the initial and corrective components of aiming movements.

This study examined whether the left and right hemispheres play differential roles in controlling the initial and corrective components of aiming movements. A simple aiming task was administered to 31 normal control subjects and 29 unilateral stroke patients (14 with right hemisphere damage and 15 with left hemisphere damage). Movement amplitude was varied (25, 64 and 100 mm) and reaction time, movement time and accuracy were measured. Through a trajectory analysis, initial and corrective movements were separated. The stroke patients performed the task with their ipsilateral arm which was compared to the normal controls' right or left are performance. Regardless of the movement amplitude the left hemisphere group's reaction time was slower, and the execution of the initial movement component was less accurate than controls. No performance deficits were found on corrective movements. Performance was not impaired for the right hemisphere group on any measures. These results are discussed in terms of the hemispheres' possible roles in controlling movements which are largely open or closed loop.

Aged↗

Complete nucleotide sequence of RNA segment 3 of bluetongue virus serotype 2 (Ona-A). Phylogenetic analyses reveal the probable origin and relationship with other orbiviruses.

The nucleotide sequence of the RNA segment 3 of bluetongue virus (BTV) serotype 2 (Ona-A) from North America was determined to be 2772 nucleotides containing a single large open reading frame of 2703 nucleotides (901 amino acid). The predicted VP3 protein exhibited general physiochemical properties (including hydropathy profiles) which were very similar to those previously deduced for other BTV VP3 proteins. Partial genome segment 3 sequences, obtained by polymerase chain reaction (PCR) sequencing, of BTV isolates from the Caribbean were compared to those from North America, South Africa, India, Indonesia, Malaysia and Australia, as well as other orbiviruses, to determine the phylogenetic relationships amongst them. Three major BTV topotypes (Gould, A.R. (1987) Virus Res. 7, 169-183) were observed which had nucleotide sequences that differed by approximately 20%. At the molecular level, geographic separation had resulted in significant divergence in the BTV genome segment 3 sequences, consistent with the evolution of distinct viral populations. The close phylogenetic relationship between the BTV serotype 2 (Ona-A strain) from Florida and the BTV serotypes 1, 6 and 12 from Jamaica and Honduras, indicated that the presence of BTV serotype 2 in North America was probably due to an exotic incursion from the Caribbean region as previously proposed by Sellers and Maaroof ((1989) Can. J. Vet. Res. 53, 100-102) based on trajectory analysis. Conversely, nucleotide sequence analysis of Caribbean BTV serotype 17 isolates suggested they arose from incursions which originated in the USA, possibly from a BTV population distinct from those circulating in Wyoming.

Amino Acid Sequence↗

Social support, social problem-solving abilities, and adjustment of family caregivers of stroke survivors.

OBJECTIVE: To determine contributions of social support and social problem-solving abilities in prediction of adjustment of family caregivers of stroke survivors. DESIGN: Descriptive. SETTING: Two rehabilitation facilities (1 private, 1 state) in the southeastern United States. PARTICIPANTS: Fifty-two family caregivers (46 women, 6 men) of stroke survivors (28 women, 24 men). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Predictor variables were social support and social problem-solving abilities. Outcome measures of caregiver adjustment were depressive symptomatology, well-being, and general health. Participants completed these measures 1 to 2 days before discharge from inpatient rehabilitation and at 5, 9, and 13 weeks postdischarge in the home. RESULTS: Trajectory analysis indicated higher levels of social support were associated with lower levels of caregiver depressive symptomatology and higher levels of well-being and general health, independent of social problem solving. A greater negative problem orientation was associated with higher levels of depressive symptomatology and lower levels of well-being. A more positive problem orientation was associated with greater increases in general health. The strength or slope of this positive relation lessened over time. CONCLUSIONS: Social support and the emotion-focused component of social problem solving, problem orientation, independently contribute to caregiver adjustment. Interventions that provide social support and assist caregivers to develop more adaptive abilities toward problem solving may be beneficial.

Adaptation, Psychological↗