Generation of calves persistently infected with HoBi-like pestivirus and comparison of methods for detection of these persistent infections.

The identification and elimination of persistently infected (PI) cattle are the most effective measures for controlling bovine pestiviruses, including bovine viral diarrhea virus (BVDV) and the emerging HoBi-like viruses. Here, colostrum-deprived calves persistently infected with HoBi-like pestivirus (HoBi-like PI calves) were generated and sampled (serum, buffy coat, and ear notches) on the day of birth (DOB) and weekly for 5 consecutive weeks. The samples were subjected to diagnostic tests for BVDV--two reverse transcriptase PCR (RT-PCR) assays, two commercial real-time RT quantitative PCR (RT-qPCR), two antigen capture enzyme-linked immunosorbent assays (ACE), and immunohistochemistry (IHC)--and to HoBi-like virus-specific RT-PCR and RT-qPCR assays. The rate of false negatives varied among the calves. The HoBi-like virus-specific RT-PCR detected HoBi-like virus in 83%, 75%, and 87% of the serum, buffy coat, and ear notch samples, respectively, while the HoBi-like RT-qPCR detected the virus in 83%, 96%, and 62%, respectively. In comparison, the BVDV RT-PCR test had a higher rate of false negatives in all tissue types, especially for the ear notch samples (missing detection in at least 68% of the samples). The commercial BVDV RT-qPCRs and IHC detected 100% of the ear notch samples as positive. While ACE based on the BVDV glycoprotein E(rns) detected infection in at least 87% of ear notches, no infections were detected using NS3-based ACE. The BVDV RT-qPCR, ACE, and IHC yielded higher levels of detection than the HoBi-like virus-specific assays, although the lack of differentiation between BVDV and HoBi-like viruses would make these tests of limited use for the control and/or surveillance of persistent HoBi-like virus infection. An improvement in HoBi-like virus tests is required before a reliable HoBi-like PI surveillance program can be designed.

Comparing the stability of diagnosis in full vs. partial posttraumatic stress disorder.

We studied differences in diagnostic stability between patients with full and patients with partial posttraumatic stress disorder (PTSD). We collected self-reported symptoms of PTSD, anxiety, depression, and functioning at a Veterans Affairs mental health clinic (n = 1962). We classified patients as meeting full or partial PTSD based upon their initial assessment. We performed Kaplan-Meier survival analysis to compare stability of diagnosis over time and Cox proportional hazards models to understand how comorbid symptoms and level of functioning confounded the relationship. We performed a chart review to examine differences in treatment received by the two groups. Patients in the partial PTSD group lost their diagnosis significantly faster and at significantly higher rates than did patients with full PTSD. Comorbid symptoms contributed significantly to this difference. Mental health treatments delivered to the two groups were similar. These diagnoses appear to be different, suggesting that people with partial PTSD may benefit from a different clinical approach.

Accumulation of Insensitive Munition Compounds in the Earthworm Eisenia andrei from Amended Soil: Methodological Considerations for Determination of Bioaccumulation Factors.

The present study investigates the bioaccumulation of the insensitive munition compounds 2,4-dinitroanisole (DNAN) and 3-nitro-1,2,4-triazol-5-one (NTO), developed for future weapons systems to replace current munitions containing sensitive explosives. The earthworm Eisenia andrei was exposed to sublethal concentrations of DNAN or NTO amended in Sassafras sandy loam. Chemical analysis indicated that 2- and 4-amino-nitroanisole (2-ANAN and 4-ANAN, respectively) were formed in DNAN-amended soils. The SumDNAN (sum of DNAN, 2-ANAN, and 4-ANAN concentrations) in soil decreased by 40% during the 14-d exposure period. The SumDNAN in the earthworm body residue increased until day 3 and decreased thereafter. Between days 3 and 14, there was a 73% decrease in tissue uptake that was greater than the 23% decrease in the soil concentration, suggesting that the bioavailable fraction may have decreased over time. By day 14, the DNAN concentration accounted for only 45% of the SumDNAN soil concentration, indicating substantial DNAN transformation in the presence of earthworms. The highest bioaccumulation factor (BAF; the tissue-to-soil concentration ratio) was 6.2 ± 1.0 kg/kg (dry wt) on day 3 and decreased to 3.8 ± 0.8 kg/kg by day 14. Kinetic studies indicated a BAF of 2.3 kg/kg, based on the earthworm DNAN uptake rate of 2.0 ± 0.24 kg/kg/d, compared with the SumDNAN elimination rate of 0.87 d-1 (half-life = 0.79 d). The compound DNAN has a similar potential to bioaccumulate from soil compared with trinitrotoluene. The NTO concentration in amended soil decreased by 57% from the initial concentration (837 mg NTO/kg dry soil) during 14 d, likely due to the formation of unknown transformation products. The bioaccumulation of NTO was negligible (BAF ≤ 0.018 kg/kg dry wt). Environ Toxicol Chem 2021;40:1713-1725. © 2021 SETAC. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

A calcium-dependent protein kinase with a regulatory domain similar to calmodulin.

Calcium can function as a second messenger through stimulation of calcium-dependent protein kinases. A protein kinase that requires calcium but not calmodulin or phospholipids for activity has been purified from soybean. The kinase itself binds calcium with high affinity. A complementary DNA clone for this kinase has been identified; it encodes a protein with a predicted molecular mass of 57,175 daltons. This protein contains a catalytic domain similar to that of calmodulin-dependent kinases and a calmodulin-like region with four calcium binding domains (EF hands). The predicted structure of this kinase explains its direct regulation via calcium binding and establishes it as a prototype for a new family of calcium-regulated protein kinases.

PlantsP: a functional genomics database for plant phosphorylation.

The PlantsP database is a curated database that combines information derived from sequences with experimental functional genomics information. PlantsP focuses on plant protein kinases and protein phosphatases. The database will specifically provide a resource for information on a collection of T-DNA insertion mutants (knockouts) in each protein kinase and phosphatase in Arabidopsis thaliana. PlantsP also provides a curated view of each protein that includes a comprehensive annotation of functionally related sequence motifs, sequence family definitions, alignments and phylogenetic trees, and descriptive information drawn directly from the literature. PlantsP is available at http://PlantsP.sdsc.edu.

CDPKs - a kinase for every Ca2+ signal?

Numerous stimuli can alter the Ca2+concentration in the cytoplasm, a factor common to many physiological responses in plant and animal cells. Calcium-binding proteins decode information contained in the temporal and spatial patterns of these Ca2+ signals and bring about changes in metabolism and gene expression. In addition to calmodulin, a calcium-binding protein found in all eukaryotes, plants contain a large family of calcium-binding regulatory protein kinases. Evidence is accumulating that these protein kinases participate in numerous aspects of plant growth and development.

Altered cortical excitability in obsessive-compulsive disorder.

OBJECTIVE: To assess cortical inhibitory and excitatory mechanisms in obsessive-compulsive disorder (OCD). BACKGROUND: Transcranial magnetic stimulation (TMS) studies have found decreased neuronal inhibition and a reduced cortical silent period in the primary motor area in Tourette's syndrome, focal dystonia, and other disorders believed to involve dysfunction of subcortical structures, including the basal ganglia. Dysfunction of the basal ganglia and linked regions also has been implicated in OCD, which has significant clinical and familial overlap with tic disorders. METHODS: We applied the TMS techniques previously used in Tourette's syndrome to a group of 16 OCD patients (seven unmedicated) and 11 age-matched healthy volunteers extensively screened for psychopathology. Measures of motor cortex excitability included resting and active motor threshold, cortical silent period duration, and intracortical inhibition and facilitation using a paired-pulse TMS technique with a subthreshold conditioning stimulus. RESULTS: Similar to recent findings in Tourette's syndrome and focal dystonia, this study reports significantly decreased intracortical inhibition (ICI) relative to the volunteers at interstimulus intervals from 2 to 5 msec. We also found decreased active and resting motor evoked potential threshold in the OCD patients, another indication of increased cortical excitability. Neither abnormality appeared medication related. The decreases in ICI and motor threshold were greatest in OCD patients with comorbid tics, but remained significant in patients without tics. CONCLUSIONS: The data suggest abnormal cortical excitability in obsessive-compulsive disorder. These findings are congruent with the hypothesis that Tourette's syndrome and obsessive-compulsive disorder (OCD) are analogous disorders with overlapping dysfunction in corticobasal circuits. Patients with tic-related OCD may have more abnormal motor cortex excitability than OCD patients without tics.

Social experience and social context alter the behavioral response to centrally administered oxytocin in female Syrian hamsters.

The type of social behavior displayed by an individual is profoundly influenced by its immediate social environment or context and its prior social experience. Although oxytocin is important in the expression of social behavior in several species, it is not known if social factors alter the ability of oxytocin to influence behavior. The purpose of the present study was to test the hypothesis that social experience and social context alter the ability of oxytocin to regulate flank marking (a form of scent marking) in female Syrian hamsters. Oxytocin was microinjected into the medial preoptic anterior hypothalamic continuum (MPOA-AH) of socially experienced, dominant female hamsters which were then tested with either a subordinate partner, with a novel partner, or alone. Oxytocin induced flank marking in a dose-dependent manner but only when the experienced dominant hamsters were tested with their familiar, subordinate partners. Oxytocin did not induce flank marking when injected into socially naive female hamsters that were tested with an opponent or alone. In males, by contrast, oxytocin induced flank marking in dominant hamsters when they were tested with their subordinate partner or alone. These data support the hypothesis that social experience and social context interact to regulate the ability of oxytocin to stimulate flank marking by its actions in the MPOA-AH in female hamsters.

Oxytocin inhibits aggression in female Syrian hamsters.

Dominant subordinate relationships are formed as the result of social conflict and are maintained at least in part by communication. At this time, little is known about the neural mechanisms that are responsible for coordinating the social behaviours (e.g. aggression) that occur in association with the formation and maintenance of these relationships. The purpose of the present study was to investigate the role of oxytocin (OXT) within the medial preoptic anterior hypothalamic continuum (MPOA-AH) in the control of aggression in female hamsters. OXT injected into the MPOA-AH immediately before testing significantly reduced the duration of aggression in a dose-dependent manner. Injection of an OXT antagonist 30 min before testing significantly increased the duration of aggression. In contrast, the duration of aggression was not altered when hamsters were tested either 30 min after injection of OXT or immediately following injection of an OXT-antagonist. These data support the hypothesis that OXT release within the MPOA-AH regulates social behaviours important in the formation and maintenance of dominant subordinate relationships in female hamsters.

Dinoflagellate sterols IV: Isolation and structure of 4 alpha, 23xi, 24xi-trimethylcholestanol from the dinoflagellate Glenodinium hallii.

The dinoflagellate Glenodinium hallii was investigated for its sterol composition. Five of the six sterols were isolated and identified as cholest-5en-3beta-ol, (24xi)-24-methylcholest-5-en-3beta-ol, stigmasta-5,22-dien-3beta-ol, (22E,24R)-4 alpha, 23, 24-trimethyl-5 alpha-cholest-22-en-3 beta-ol, and 4 alpha, 23 xi, 24 xi-trimethyl-5 alpha-cholestan-3 beta-ol.

A cost analysis of a tele-oncology practice.

Costs were monitored for three different types of oncology practice: a telemedicine clinic and a fly-in outreach clinic, both held in rural areas, and a traditional clinic held in a city hospital. Total expenses were calculated over the year May 1995 to April 1996. The average cost per telemedicine visit was $812. The average cost per outreach clinic visit was $897. Flying in oncology support for this practice was therefore about 10% more costly than telemedicine. While the outreach cost may have been inappropriately high due to a slow start-up phase, it was still less expensive during this period to be seen via telemedicine. For comparison, the average cost per traditional oncology clinic visit was $149. However, this figure does not take into account the costs of access to a city-based service by rural patients.

A cost measurement study for a tele-oncology practice.

The costs of providing oncology services in three different ways were measured. Services were provided to a peripheral hospital by: conventional clinics, in which the oncologist worked at the hospital concerned; outreach clinics, in which an oncologist was flown in periodically from a central hospital; telemedicine clinics, in which the oncologist at the central hospital practised via a video-link. During a one-year study period, 2400 patients were seen in conventional clinics, 81 in outreach clinics and 103 in telemedicine clinics. At these workloads the average costs per patient were $149, $897 and $812, respectively. However, the average costs cannot be compared directly without further information about the shape of the unit cost curves.

Preliminary findings for a brief posttraumatic stress intervention in primary mental health care.

A team of clinicians at a small rural Veterans' Health Administration (VHA) medical center piloted a brief psychological intervention for posttraumatic stress in a primary mental health care setting. Symptom measures were completed by veterans before and after receiving the brief trauma treatment (BTT), and were then analyzed using paired t tests. In our uncontrolled study, we found a statistically insignificant improvement in symptoms of posttraumatic stress disorder, though there were statistically significant, but not clinically significant, improvements in depression and general anxiety. The intervention may enhance subsequent specialty mental health engagement. Fifty-one veterans (62.20%) went on to receive psychotherapy in a specialty mental health setting, which represents a substantial increase in specialty psychotherapy engagement compared to reports elsewhere in the literature. Lack of controlled comparison precludes definitive conclusions, but the current preliminary results support future studies of brief psychological interventions in primary care settings, including randomized controlled comparisons.

Immunolocalization of retinoic acid receptor-alpha, -beta, and -gamma, in bovine and canine sperm.

Retinoic acid is an important regulator of cellular proliferation and differentiation. The action of retinoic acid is mediated by retinoic acid receptors (RARs) and the retinoid X receptors. The objective was to elucidate the protein localization and expression of RARα, RARß, and RARγ in bull and dog sperm. Bull and dog sperm were subjected to an immunostaining procedure to determine presence of RARα, RARß, and RARγ. We concluded that all three receptors were present in different regions of bull and dog sperm at varying levels. Protein expression in bull and dog sperm lysates was investigated using protein dot-blot analyses. The protein levels of RARα and RARγ were higher than the protein level of RARß in bull and dog sperm. Protein sequences of RARα, RARß, and RARγ for bull and dog were 98%, 89%, and 98%, respectively, on similarity alignment. In conclusion, the presence of RARα, RARß, and RARγ receptors supported their role in sperm structure and function.

Effect of extenders on sperm mitochondrial membrane, plasma membrane and sperm kinetics during liquid storage of canine semen at 5°C.

The objective of this study was to evaluate the effect of addition of egg yolk (EY) or soy lecithin (SL) based extenders on dog sperm parameters during 10days storage at 5°C. Four ejaculates of pooled semen from three Mongrel dogs were divided into three aliquots and extended to a final concentration of 200 million sperm/mL using following non-commercial extenders: egg yolk extender (EYE) group - semen+20% Tris-EYE; soy lecithin (SL) group 1 - semen+SL extender with 0.04% SL; SL group 2 - semen+SL extender 0.4% SL. The extended semen samples were stored at 5°C and were evaluated for sperm mitochondrial membrane potential (MMP), plasma membrane integrity (%PMI) and computer assisted sperm analyzer (CASA) sperm motility parameters on days 0, 1, 2, 3, 4, 7 and 10. The extender, days of storage and extender×days of storage affected the MMP, %PMI and CASA sperm motility parameters (P<0.05). The addition of 0.4% SL extender reduced the speed of deterioration of sperm parameters evaluated in this study compared to EY and 0.04% SL extenders (P<0.05). In conclusion, the addition of 0.4% SL extender enhanced sperm preservation based on the variables evaluated in the present study compared to EYE and 0.04% SL extenders and plausibly preserves sperm quality longer.

Characterization of silver nanoparticles using flow-field flow fractionation interfaced to inductively coupled plasma mass spectrometry.

The ability to detect and identify the physiochemical form of contaminants in the environment is important for degradation, fate and transport, and toxicity studies. This is particularly true of nanomaterials that exist as discrete particles rather than dissolved or sorbed contaminant molecules in the environment. Nanoparticles will tend to agglomerate or dissolve, based on solution chemistry, which will drastically affect their environmental properties. The current study investigates the use of field flow fractionation (FFF) interfaced to inductively coupled plasma-mass spectrometry (ICP-MS) as a sensitive and selective method for detection and characterization of silver nanoparticles. Transmission electron microscopy (TEM) is used to verify the morphology and primary particle size and size distribution of precisely engineered silver nanoparticles. Subsequently, the hydrodynamic size measurements by FFF are compared to dynamic light scattering (DLS) to verify the accuracy of the size determination. Additionally, the sensitivity of the ICP-MS detector is demonstrated by fractionation of µg/L concentrations of mixed silver nanoparticle standards. The technique has been applied to nanoparticle suspensions prior to use in toxicity studies, and post-exposure biological tissue analysis. Silver nanoparticles extracted from tissues of the sediment-dwelling, freshwater oligochaete Lumbriculus variegatus increased in size from approximately 31-46nm, indicating a significant change in the nanoparticle characteristics during exposure.

Paradoxical vocal cord motion--a dangerous imitator of airway emergencies.

We report a case of stridor in a 32 year old woman. Initial laryngoscopy demonstrated adduction of the vocal cords on inspiration, which reverted to abduction on induction of general anaesthesia. The airway was structurally normal. The most likely diagnosis was paradoxical vocal cord motion, a condition in which psychological stress can precipitate respiratory symptoms and signs due to involuntary adduction of the vocal cords during inspiration. Its importance to the anaesthetist lies in its ability to masquerade as a serious airway or respiratory condition.

Cancer prevention among rural youth: building a "bridge" to better health with genealogy.

BACKGROUND: The Bridge to Better Health (BRIDGE) project is a program that focuses on providing rural high school youth with motivation, knowledge, and skills essential to cancer prevention. In this pilot intervention, we used instruction in personal health genealogy as a means of increasing awareness and knowledge of health risk and motivation to change several screening and cancer-related behaviors. METHODS: We administered a Bridge to Better Health survey to 173 ninth- and 10th-grade students from a rural Southeastern Virginia high school before and after delivery of the BRIDGE pilot intervention. RESULTS: Significant preintervention to postintervention changes were observed for general genealogy knowledge, personal health genealogy, self-efficacy, and intention to practice self-examinations (breast, testicular, and skin) and eating a high-fiber and low-fat diet. CONCLUSION: These project results demonstrate the importance of theory-driven interventions for increasing cancer knowledge and changing cancer-related dietary and screening behaviors.

Mechanical cardiopulmonary resuscitation.

Mechanical cardiopulmonary resuscitation is used by prehospital and intrahospital personnel to provide basic and advanced life support. It has several advantages over manual CPR. Major advances have been made to produce convenient portable units that provide reliable, improved ventilation and cardiac compression. More sophisticated techniques under development would effect changes in prehospital emergency care procedures because they require endotracheal intubation. Mechanical CPR will logically be important in implementing these techniques.