Characterization of a bovine intestinal myofibroblast cell line and stimulation using phytoglycogen-based nanoparticles bound to inosine monophosphate.

The goal of the present study was to characterize a novel bovine intestinal myofibroblast (BT-IMF) cell line isolated from a fetal bovine intestine. This cell type is of importance as intestinal myofibroblasts play a key role in controlling intestinal epithelial cell proliferation, intestinal regulation, wound healing, epithelial cell turnover, and structural support. The present work demonstrates that BT-IMF cells could be successfully cryopreserved and thawed and cultured past 25 passages. Immunocytochemical staining of the BT-IMF cell line was positive for vimentin and smooth muscle actin (α-SMA) and negative for pancytokeratin, suggesting that the cells are myofibroblastic in type. Growth kinetic experiments demonstrate that hydrocortisone negatively impacts BT-IMF growth and non-essential amino acids enhance its proliferation. Inosine monophosphate (IMP) is a dietary nucleotide and is essential for supporting animal health. Stimulation with IMP bound to a novel phytoglycogen-based nanocarrier (IMP-NP) showed enhanced cell proliferation. BT-IMF provides a new tool for studying bovine cells in vitro and may be of particular interest for cultured meat manufacturing in the future.

A pilot study augmenting cognitive behavioral therapy for panic disorder with attention bias modification: Clinical and psychophysiological outcomes.

BACKGROUND AND OBJECTIVES: Studies show that attentional bias towards threat is a key maintenance factor for panic disorder (PD). Attentional bias may be an important mechanism of symptom reduction, and thus, a useful target for optimizing outcomes. The current study examined whether an attention bias modification (ABM) task enhanced CBT outcomes. Multiple methods for assessing PD were used, including physiological measurement and clinician-rated assessment. METHODS: Adults with panic disorder (N = 24) received seven sessions of CBT with either ABM or sham attention tasks. Psychophysiological reaction to a loud tones startle paradigm was assessed before and after treatment. RESULTS: Across both groups, panic symptom severity decreased with CBT. The ABM group showed greater reductions in PD symptoms compared to the placebo group. Notably, however, changes in attentional bias were not associated with symptom reductions across groups. No significant group differences on psychophysiological assessment were observed. LIMITATIONS: This study is limited by the small sample size, which rendered our power to investigate mediators and moderators insufficient. More research is needed to validate the effect of attention bias modification on attentional bias to threat. CONCLUSIONS: This pilot study suggests that future research should investigate attentional bias in conjunction with CBT. A larger sample would provide opportunity to further investigate the mechanisms through which ABM works, along with potential moderating factors and the use of psychophysiological measurements in panic disorder.

Extracellular dsRNA induces a type I interferon response mediated via class A scavenger receptors in a novel Chinook salmon derived spleen cell line.

Despite increased global interest in Chinook salmon aquaculture, little is known of their viral immune defenses. This study describes the establishment and characterization of a continuous cell line derived from Chinook salmon spleen, CHSS, and its use in innate immune studies. Optimal growth was seen at 14-18 °C when grown in Leibovitz's L-15 media with 20% fetal bovine serum. DNA analyses confirmed that CHSS was Chinook salmon and genetically different from the only other available Chinook salmon cell line, CHSE-214. Unlike CHSE-214, CHSS could bind extracellular dsRNA, resulting in the rapid and robust expression of antiviral genes. Receptor/ligand blocking assays confirmed that class A scavenger receptors (SR-A) facilitated dsRNA binding and subsequent gene expression. Although both cell lines expressed three SR-A genes: SCARA3, SCARA4, and SCARA5, only CHSS appeared to have functional cell-surface SR-As for dsRNA. Collectively, CHSS is an excellent cell model to study dsRNA-mediated innate immunity in Chinook salmon.

PTSD in women is associated with a block in conversion of progesterone to the GABAergic neurosteroids allopregnanolone and pregnanolone measured in plasma.

There is a need to identify new and more effective treatments for posttraumatic stress disorder (PTSD). Allopregnanolone and its stereoisomer pregnanolone (together termed ALLO) are metabolites of progesterone that positively and allosterically modulate GABA effects at GABAA receptors, thereby reducing anxiety and depression. Previous research revealed that women with PTSD had low cerebrospinal fluid (CSF) ALLO levels and a low ratio of ALLO to the allopregnanolone precursor 5α-DHP, consistent with deficient activity of the ALLO synthetic enzyme 3α-hydroxysteroid dehydrogenase (3α-HSD). The current study examined ALLO and the ratio of ALLO to 5α-DHP in plasma at rest and in response to psychophysiological stressors in trauma-exposed, medication-free women with and without PTSD. Participants were examined twice in random order during the early follicular phase (eFP) and mid-luteal phase (mLP) of the menstrual cycle. Plasma neurosteroids were measured using gas chromatography-mass spectrometry. Results indicate that the ALLO to 5α-DHP ratio in plasma increases between the eFP and mLP. In addition, women with PTSD have a lower ratio of ALLO to 5α-DHP than trauma-exposed healthy women, as well as blunted increases in this ratio in response to a moderately stressful laboratory procedure, i.e., differential fear conditioning, across the menstrual cycle. Clinically feasible testing for 3α-HSD dysfunction is critical to translating this line of research into clinical care. Measurement of this ratio in plasma could facilitate patient stratification in clinical treatment trials, as well as precision medicine targeting of treatments that address ALLO synthesis deficits in women with PTSD.

Identification of three class A scavenger receptors from rainbow trout (Oncorhynchus mykiss): SCARA3, SCARA4, and SCARA5.

Class A scavenger receptors (SR-As) are a family of five surface receptors whose functions in mammals are associated with innate immunity; however, their role in fish immunity requires further elucidation. The present study identifies, performs sequence analysis, and constitutive transcript expression analysis for three SR-A family members, SCARA3, SCARA4 and SCARA5, from rainbow trout. This work will provide a basis for future studies on SR-A function and their role in innate immunity in this economically important fish.

Diminished medial prefrontal cortex activation during the recollection of stressful events is an acquired characteristic of PTSD.

BACKGROUND: Previous research has shown relatively diminished medial prefrontal cortex activation and heightened psychophysiological responses during the recollection of personal events in post-traumatic stress disorder (PTSD), but the origin of these abnormalities is unknown. Twin studies provide the opportunity to determine whether such abnormalities reflect familial vulnerabilities, result from trauma exposure, or are acquired characteristics of PTSD. METHODS: In this case-control twin study, 26 male identical twin pairs (12 PTSD; 14 non-PTSD) discordant for PTSD and combat exposure recalled and imagined trauma-unrelated stressful and neutral life events using a standard script-driven imagery paradigm during functional magnetic resonance imaging and concurrent skin conductance measurement. RESULTS: Diminished activation in the medial prefrontal cortex during Stressful v. Neutral script-driven imagery was observed in the individuals with PTSD, relative to other groups. CONCLUSIONS: Diminished medial prefrontal cortex activation during Stressful v. Neutral script-driven imagery may be an acquired characteristic of PTSD. If replicated, this finding could be used prospectively to inform diagnosis and the assessment of treatment response.

CHSE-214: A model for studying extracellular dsRNA sensing in vitro.

Double-stranded RNA (dsRNA) is produced by almost all viruses during their replicative cycle and is a potent inducer of the innate antiviral immune response including inducing expression of type I interferons (IFNs) and interferon-stimulated genes (ISGs). During lytic virus infections intracellular dsRNA can escape into the extracellular space, where surface pattern recognition receptors, such as class A scavenger receptors (SR-As) facilitate its binding and entry into neighbouring cells. Studying extracellular dsRNA entry is difficult due to the ubiquitous expression profile and compensatory dsRNA binding characteristics of SR-As; a SR-A deficient cell line has yet to be identified. The present study suggests the Chinook salmon embryonic cell line, CHSE-214, as a model for studying extracellular dsRNA sensing in vitro. CHSE-214 is unable to bind and respond to extracellular dsRNA, can only respond to dsRNA when it is transfected into the cells, and is able to bind dsRNA when overexpressing human SR-AI. The applications for this model could include elucidating: dsRNA binding and entry mechanisms, including sequence and length effects, as well as SR-A and other putative surface dsRNA receptor ligand binding studies.

Formation of Defence Primary Healthcare: a new way of delivering firm base primary healthcare.

Following the Strategic Defence and Security Review of 2010, the UK Surgeon General was directed to merge the delivery of primary healthcare from the three single Service organisations to a unified Defence Primary Healthcare. Although front line clinical staff were to be preserved, considerable savings were to be made in headquarters staff. This was one of the largest UK military medicine changes in delivery for a generation. The changes were completed on time with the transfer of UK and overseas general practice, specialist community services and dentistry, with a later requirement to add healthcare for the Reserves. The first years of this initiative have been remarkably successful, and Defence Primary Healthcare (DPHC) has progressively increased performance in all the QOF criteria measured by Defence Statistics.

Small structural changes of the imidazopyridine diacylglycerol acyltransferase 2 (DGAT2) inhibitors produce an improved safety profile.

Small molecule DGAT2 inhibitors have shown promise for the treatment of metabolic diseases in preclinical models. Herein, we report the first toxicological evaluation of imidazopyridine-based DGAT2 inhibitors and show that the arteriopathy associated with imidazopyridine 1 can be mitigated with small structural modifications, and is thus not mechanism related.

The clinical nurse specialist's role in head and neck cancer care: United Kingdom National Multidisciplinary Guidelines.

This is the official guideline endorsed by the specialty associations involved in the care of head and neck cancer patients in the UK. It discusses the role of the clinical nurse specialist in the head and neck cancer patient journey and provides recommendations on the clinical nurse specialist led assessments and interventions for this group of patients receiving cancer care. Recommendations • All cancer patients should meet a clinical nurse specialist at the point of diagnosis. (R) • Clinical nurse specialists must act as gate keeper to the patients' cancer pathway to provide a seamless journey. (R) • Holistic needs assessment should be completed at different stages of the patient's pathway to reflect the changes of the patients' needs. (R) • Clinical nurse specialists to be part of local and national initiatives for health promotion and raising awareness in the public domain. (G) • Clinical nurse specialists should lead in redesigning of services and policies to ensure they are responsive to patient's needs for the future. (G) • Treatment summaries should become part of practice to provide good communication between primary and secondary care to enable continuity of care for the patient. (G).

Single electron transfer (SET) activity of the dialkyl-amido sodium zincate [(TMEDA)·Na(µ-TMP)(µ-tBu)Zn(tBu)] towards TEMPO and chalcone.

More usually thought of as a base, the sodium zincate [(TMEDA)·Na(µ-TMP)(µ-(t)Bu)Zn((t)Bu)] 1 can undergo single electron transfer with TEMPO to give [(TMEDA)·Na(µ-TMP)(µ-TEMPO(-))Zn((t)Bu)] 2 and [(TMEDA)·Na(µ-TEMPO(-))(2)Zn((t)Bu)] 3; and with chalcone [PhCOCH=CHPh] gives [{(TMEDA)·Na(µ-TMP)Zn((t)Bu)}(2)(µ-OCPhCH=CHPhCHPhCH=CPh-µ-O)] which contains two chalcone units C-C coupled though their benzylic C atoms.

Decreased regional cerebral blood flow in medial prefrontal cortex during trauma-unrelated stressful imagery in Vietnam veterans with post-traumatic stress disorder.

BACKGROUND: Neuroimaging research has demonstrated medial prefrontal cortex (mPFC) hyporesponsivity and amygdala hyperresponsivity to trauma-related or emotional stimuli in post-traumatic stress disorder (PTSD). Relatively few studies have examined brain responses to the recollection of stressful, but trauma-unrelated, personal events in PTSD. In the current study, we sought to determine whether regional cerebral blood flow (rCBF) abnormalities in mPFC and amygdala in PTSD could be observed during the recollection of trauma-unrelated stressful personal events. METHOD: Participants were 35 right-handed male combat veterans (MCVs) and female nurse veterans (FNVs) who served in Vietnam: 17 (seven male, 10 female) with current military-related PTSD and 18 (nine male, nine female) with no current or lifetime PTSD. We used positron emission tomography (PET) and script-driven imagery to study rCBF during the recollection of trauma-unrelated stressful versus neutral and traumatic events. RESULTS: Voxelwise tests revealed significant between-group differences for the trauma-unrelated stressful versus neutral comparison in mPFC, specifically in the anterior cingulate cortex (ACC). Functional region of interest (ROI) analyses demonstrated that this interaction in mPFC represented greater rCBF decreases in the PTSD group during trauma-unrelated stressful imagery relative to neutral imagery compared to the non-PTSD group. No differential amygdala activation was observed between groups or in either group separately. CONCLUSIONS: Veterans with PTSD, compared to those without PTSD, exhibited decreased rCBF in mPFC during mental imagery of trauma-unrelated stressful personal experiences. Functional neuroanatomical models of PTSD must account for diminished mPFC responses that extend to emotional stimuli, including stressful personal experiences that are not directly related to PTSD.

Reducing MCM levels in human primary T cells during the G(0)-->G(1) transition causes genomic instability during the first cell cycle.

DNA replication is tightly regulated, but paradoxically there is reported to be an excess of MCM DNA replication proteins over the number of replication origins. Here, we show that MCM levels in primary human T cells are induced during the G(0)-->G(1) transition and are not in excess in proliferating cells. The level of induction is critical as we show that a 50% reduction leads to increased centromere separation, premature chromatid separation (PCS) and gross chromosomal abnormalities typical of genomic instability syndromes. We investigated the mechanisms involved and show that a reduction in MCM levels causes dose-dependent DNA damage involving activation of ATR & ATM and Chk1 & Chk2. There is increased DNA mis-repair by non-homologous end joining (NHEJ) and both NHEJ and homologous recombination are necessary for Mcm7-depleted cells to progress to metaphase. Therefore, a simple reduction in MCM loading onto DNA, which occurs in cancers as a result of aberrant cell cycle control, is sufficient to cause PCS and gross genomic instability within one cell cycle.

An improved water footprint methodology linking global consumption to local water resources: a case of Spanish tomatoes.

A water footprint (WF) measures the total water consumed by a nation, business or individual by calculating the total water used during the production of goods and services. This paper extends the existing methods for WF to more localised levels for crops grown partly in open systems and partly in plastic-covered houses with multi-seasonal harvesting, such as the horticulture industry in Spain. This improvement makes it possible to visualise the links of EU tomato consumption to precise production sites in Spain and opens a debate to the usefulness of such findings. This paper also compares existing ecological methodologies with WF and argues that both life cycle analysis (LCA) and ecological footprint (EF) models could benefit from WF methods. Our results show that the EU consumes 957,000 tons of Spanish fresh tomatoes annually, which evaporates 71 Mm(3)/yr of water and would require 7 Mm(3)/yr of water to dilute leached nitrates in Spain. In Spain, tomato production alone evaporates 297 Mm(3)/yr and pollutes 29 Mm(3)/yr of freshwater. Depending upon the local agro-climatic character, status of water resources, total tomato production volumes and production system, the impact of EU consumption of fresh tomatoes on Spanish freshwater is very location specific. The authors suggest that business now seek to report and address negative impacts on the environment. WF opens the door to complex water relationships and provides vital information for policy actors, business leaders, regulators and managers to their draw, dependence and responsibilities on this increasingly scarce resource.

Ensuring the success of local public health workforce assessments: using a participatory-based research approach with a rural population.

OBJECTIVES: To investigate a process for comprehensive rural public health workforce data collection, and apply this process to a competency and training needs assessment of local health department (LHD) workers in the state of Kansas, USA. STUDY DESIGN: Participatory research methods were used to determine an appropriate process for data collection. Survey instruments included the Council on Linkages public health core competencies and Columbia University public health emergency preparedness competencies. METHODS: LHD workers collaborated with the state health department to develop and pre-test training for LHD directors about the nature and purpose of the survey, as well as instructions for distributing it to their staff members. The final survey instrument included demographics, a workforce competency assessment, and an assessment of training interests, motivators and barriers. Surveys were stratified by occupational type, with employees in professional roles asked to report on additional competencies. RESULTS: All 1501 Kansas LHD employees received the needs assessment survey, and 1141 (76%) were returned. Respondents reported greater mean 'importance to job' than ability across competency domains, indicating potential training needs. Across occupational types, primary training motivators were increased competency and personal satisfaction. Barriers included lack of time, cost and family commitments. CONCLUSIONS: Using participatory research methods, the state of Kansas was able to achieve a high response rate from LHD workers. This process can serve as a model for other rural communities and organizations with limited resources. In addition, the survey results provide information about competency-oriented knowledge and training gaps of sectors of the local public health workforce, which can be used to develop training in a targeted fashion.

Induction of homotypic aggregation in the rainbow trout macrophage-like cell line, RTS11.

The rainbow trout monocyte/macrophage-like cell line, RTS11, has been used to study homotypic aggregation (HA), which is a well-studied feature of leucocytes in mammals but less understood in fish. HA is the aggregation of cells of the same cell type. RTS11 underwent HA in response to polyinosinic:cytidylic acid (poly IC), polyadenylic acid (poly A), lipopolysaccharide (LPS), zymosan, and phorbol 12-myristate 13-acetate (PMA). Poly IC was the best inducer of HA and did so in a dose- and time-dependent manner. The induction of RTS11 aggregation by poly IC required divalent cations but was not blocked by either an inhibitor of lymphocyte function-associated molecule-1 (LFA-1) or the tripeptide integrin adhesion recognition sequence, RGD. Poly IC-induced HA was inhibited by colchicine and latrunculin B, which act on microtubules and microfilaments, respectively, implying the necessity for an intact cytoskeleton. HA induction by poly IC did not occur at 4 degrees C and was blocked by the transcriptional and translational inhibitors, actinomycin D and cycloheximide, respectively, suggesting the requirement for de novo protein synthesis. Poly IC-induced RTS11 aggregation was blocked by two inhibitors of dsRNA-dependent protein kinase (PKR). This is the first indication that PKR could have a role in the HA of leucocytes.

Development of a continuous cell line, PBLE, from an American eel peripheral blood leukocyte preparation.

A continuous cell line, PBLE, was developed from the adherent cells in a culture of peripheral blood leukocytes from the American eel, Anguilla rostrata. The cells were grown in Leibovitz's L-15 basal medium supplemented with 20% fetal bovine serum (FBS). Under normal culture conditions at 18 degrees C, the morphology of PBLE was fibroblast-like. The cultures have been subcultured over 80 times and have been cryopreserved successfully. These cells have a diploid karyotype of 38 chromosomes, survived temperatures from 5 to 36 degrees C, and proliferated at temperatures from 5 degrees C to at least 30 degrees C. PBLE underwent apoptosis in response to gliotoxin, but did not show a respiratory burst. Results suggest that PBLE may have arisen from a circulating mesenchymal stem cell. PBLE was susceptible to Chum salmon reovirus (CSV) and supported CSV replication. Therefore this cell line should be useful in studying eel specific virus-host interactions.

Sex-specific influences of vasopressin on human social communication.

Arginine vasopressin (AVP) and related peptides affect social behaviors in numerous species, but AVP influences on human social functions have not yet been established. Here, we describe how intranasal AVP administration differentially affects social communication in men and women, and we propose a mechanism through which it may exert those influences. In men, AVP stimulates agonistic facial motor patterns in response to the faces of unfamiliar men and decreases perceptions of the friendliness of those faces. In contrast, in women, AVP stimulates affiliative facial motor patterns in response to the faces of unfamiliar women and increases perceptions of the friendliness of those faces. AVP also affected autonomic responsiveness to threatening faces and increased anxiety, which may underlie both communication patterns by promoting different social strategies in stressful contexts in men and women.