The Nasal Solitary Chemosensory Cell Signaling Pathway Triggers Mouse Avoidance Behavior to Inhaled Nebulized Irritants.

The nasal epithelium houses a population of solitary chemosensory cells (SCCs). SCCs express bitter taste receptors and taste transduction signaling components and are innervated by peptidergic trigeminal polymodal nociceptive nerve fibers. Thus, nasal SCCs respond to bitter compounds, including bacterial metabolites, and these reactions evoke protective respiratory reflexes and innate immune and inflammatory responses. We tested whether SCCs are implicated in aversive behavior to specific inhaled nebulized irritants using a custom-built dual-chamber forced-choice device. The behavior of mice was recorded and analyzed for the time spent in each chamber. Wild-type (WT) mice exhibited an aversion to 10 mm denatonium benzoate (Den) or cycloheximide and spent more time in the control (saline) chamber. The SCC-pathway knock-out (KO) mice did not exhibit such an aversion response. The bitter avoidance behavior of WT mice was positively correlated with the concentration increase of Den and the number of exposures. Bitter-ageusic P2X2/3 double KO mice similarly showed an avoidance response to nebulized Den, excluding the taste system's involvement and pointing to an SCC-mediated major contributor to the aversive response. Interestingly, SCC-pathway KO mice showed an attraction to higher Den concentrations; however, chemical ablation of the olfactory epithelium eliminated this attraction attributed to the smell of Den. These results demonstrate that activation of SCCs leads to a rapid aversive response to certain classes of irritants with olfaction, but not gustation, contributing to the avoidance behavior during subsequent irritant exposures. This SCC-mediated avoidance behavior represents an important defense mechanism against the inhalation of noxious chemicals.

Prolonged Amphetamine Exposures Increase the Endogenous Human Dopamine Receptors 2 at the Cellular Membrane in Cells Lacking the Dopamine Transporter.

The dopamine 2 receptors (D2R) are G-protein coupled receptors expressed both in pre- and post-synaptic terminals that play an important role in mediating the physiological and behavioral effects of amphetamine (Amph). Previous studies have indicated that the effects of Amph at the D2R mainly rely on the ability of Amph to robustly increase extracellular dopamine through the dopamine transporter (DAT). This implies that the effects of Amph on D2R require the neurotransmitter dopamine. However, because of its lipophilic nature, Amph can cross the cellular membrane and thus potentially affect D2R expression independently of dopamine and DAT, e.g., in post-synaptic terminals. Here we used an in vitro system to study whether Amph affects total expression, cellular distribution, and function of the human D2R (hD2R), endogenously expressed in HEK293 cells. By performing Western blot experiments, we found that prolonged treatments with 1 or 50 µM Amph cause a significant decrease of the endogenous hD2R in cells transfected with human DAT (hDAT). On the other hand, in cells lacking expression of DAT, quantification of the hD2R-mediated changes in cAMP, biotinylation assays, Western blots and imaging experiments demonstrated an increase of hD2R at the cellular membrane after 15-h treatments with Amph. Moreover, imaging data suggested that barbadin, a specific inhibitor of the ßarrestin-ßadaptin interaction, blocked the Amph-induced increase of hD2R. Taken together our data suggest that prolonged exposures to Amph decrease or increase the endogenous hD2R at the cellular membrane in HEK293 cells expressing or lacking hDAT, respectively. Considering that this drug is often consumed for prolonged periods, during which tolerance develops, our data suggest that even in absence of DAT or dopamine, Amph can still alter D2R distribution and function.

Robotic Assay for Drought (RoAD): an automated phenotyping system for brassinosteroid and drought responses.

Brassinosteroids (BRs) are a group of plant steroid hormones involved in regulating growth, development, and stress responses. Many components of the BR pathway have previously been identified and characterized. However, BR phenotyping experiments are typically performed in a low-throughput manner, such as on Petri plates. Additionally, the BR pathway affects drought responses, but drought experiments are time consuming and difficult to control. To mitigate these issues and increase throughput, we developed the Robotic Assay for Drought (RoAD) system to perform BR and drought response experiments in soil-grown Arabidopsis plants. RoAD is equipped with a robotic arm, a rover, a bench scale, a precisely controlled watering system, an RGB camera, and a laser profilometer. It performs daily weighing, watering, and imaging tasks and is capable of administering BR response assays by watering plants with Propiconazole (PCZ), a BR biosynthesis inhibitor. We developed image processing algorithms for both plant segmentation and phenotypic trait extraction to accurately measure traits including plant area, plant volume, leaf length, and leaf width. We then applied machine learning algorithms that utilize the extracted phenotypic parameters to identify image-derived traits that can distinguish control, drought-treated, and PCZ-treated plants. We carried out PCZ and drought experiments on a set of BR mutants and Arabidopsis accessions with altered BR responses. Finally, we extended the RoAD assays to perform BR response assays using PCZ in Zea mays (maize) plants. This study establishes an automated and non-invasive robotic imaging system as a tool to accurately measure morphological and growth-related traits of Arabidopsis and maize plants in 3D, providing insights into the BR-mediated control of plant growth and stress responses.

A Tiered Framework for Organizing and Categorizing Medical Interoperability.

Developing, cultivating, and sustaining medical interoperability strengthens the support we provide to the warfighter by presenting our Commanders options and efficiencies to the way we can enable their operations. As our national security and defense strategies change the way our forces are employed to address our security risks throughout the world, some military commands will find they cannot provide adequate medical care without working in concert with willing and available partners.This article proposes a tiered framework that allows medical personnel to further describe and organize their engagement activities around the concept and practicalities of medical interoperability. As resources become diverted to other theaters or missions expand beyond assigned capabilities, medical interoperability provides Commanders with options to medically enable their missions through their partnerships with others. This framework links and connects activities and engagements to build partner capacity with long-term or regional interoperability among our partners and challenges engagement planners to consider ways to build interoperability at all four tiers when planning or executing health engagements and global health development. Using this framework when planning or evaluating an engagement or training event will illuminate opportunities to develop interoperability that might have otherwise been unappreciated or missed.

Early-life immune activation increases song complexity and alters phenotypic associations between sexual ornaments.

Early-life adversity can have long-lasting effects on physiological, behavioural, cognitive, and somatic processes. Consequently, these effects may alter an organism's life-history strategy and reproductive tactics.In response to early-life immune activation, we quantified levels of the acute phase protein haptoglobin (Hp) during development in male zebra finches (Taeniopygia guttata). Then, we examined the long-term impacts of early-life immune activation on an important static sexual signal, song complexity, as well as effects of early-life immune activation on the relationship between song complexity and a dynamic sexual signal, beak colouration. Finally, we performed mate-choice trials to determine if male early-life experience impacted female preference.Challenge with keyhole limpet hemocyanin (KLH) resulted in increased song complexity compared to lipopolysaccharide (LPS) treatment or the control. Hp levels were inversely correlated with song complexity. Moreover, KLH-treatment resulted in negative associations between the two sexual signals (beak colouration and song complexity). Females demonstrated some preference for KLH-treated males over controls and for control males over LPS-treated males in mate choice trials.Developmental immune activation has variable effects on the expression of secondary sexual traits in adulthood, including enhancing the expression of some traits. Because developmental levels of Hp and adult song complexity were correlated, future studies should explore a potential role for exposure to inflammation during development on song learning.Early-life adversity may differentially impact static versus dynamic signals. The use of phenotypic correlations can be a powerful tool for examining the impact of early-life experience on the associations among different traits, including sexual signals.

Teaching Biopsychosocial Competence and the Principles of the Primary Health Care (PHC) at the Patient's Bedside

The importance of behavioural and social determinants in health was recognised long ago; yet we still grapple with the challenges of developing appropriate teaching pedagogies to bring these principles into routine clinical practice. A teaching pedagogy blending the biopsychosocialapproach and the principles of primary health care (PHC); as expressed in the Alma-Ata Declaration of 1978; is lacking in the literature. This report hopes to address this need.In 1994 the University of Cape Town (UCT); South Africa; adopted a PHC-based approach to health sciences education to equip its graduates with the necessary knowledge; skills and attributes required to meet the challenges of providing health care in a country with vast socio-political inequalities. This paper describes an educational pedagogy which weaves these principles into clinical practice in an undergraduate medical clerkship. The methodology uses real patient encounters linked to an interactive seminar and a portfolio of case studies. Students described the teaching pedagogy as interesting and informative. They recognised the importance of holistic; patient-centeredcare based on a biopsychosocial approach and the importance of the PHC principles. Barriers to implementing this approach were also highlighted. The pedagogy; in use for four years; is being adopted by another department; indicating the sustainability and success of the course

Teaching biopsychosocial competence and the principles of primary health care (PHC) at the patient's bedside

The importance of behavioural and social determinants in health was recognised long ago; yet we still grapple with the challenges of developing appropriate teaching pedagogies to bring these principles into routine clinical practice. A teaching pedagogy blending the biopsychosocial appro- oach and the principles of primary health care (PHC); as expressed in the Alma-Ata Declaration of 1978; is lacking in the literature. This report hopes to address this need.In 1994 the University of Cape Town (UCT); South Africa; adopted a PHC-based approach to health sciences education to equip its graduates with the necessary knowledge; skills and attributes required to meet the challenges of providing health care in a country with vast socio-political inequalities. This paper describes an educational pedagogy which weaves these principles into clinical practice in an undergraduate medical clerkship. The methodology uses real patient encounters linked to an interactive seminar and a portfolio of case studies.Students described the teaching pedagogy as interesting and informative. They recognised the importance of holistic; patient-centered care based on a biopsychosocial approach and the importance of the PHC principles. Barriers to implementing this approach were also highlighted. The pedagogy; in use for four years; is being adopted by another department; indicating the sustainability and success of the course

Relationships, trust, decision-making and quality of care in a paediatric intensive care unit.

OBJECTIVE: To assess the care-giving practices of health-care practitioners in the paediatric intensive care unit (PICU) through their qualitative insights, reflections and experience in participatory action research. DESIGN AND METHODS: Qualitative research in the form of 'participatory action research' was used to gather data from three sources within the unit: focus groups within disciplines, observations within the PICU, and semi-structured interviews. All staff members were active collaborators and equal stakeholders in the decision-making process, research and feedback. SETTING: The paediatric intensive care unit (PICU) of the Red Cross War Memorial Children's Hospital (RCWMCH). PARTICIPANTS: All staff members from various disciplines working in the PICU. RESULTS: Staff members described problems with respect to relationships, trust and decision-making within care-giving practices. CONCLUSION: The study qualitatively describes how poor communication amongst staff members in respect of relationships and decision-making impacted on trust and how this tended to compromise care-giving practices in the PICU. The data suggested that this was more evident in informal rather than formal clinical decision-making procedures. The strength of the study was that the participatory action design in the research allowed staff members to address the very dynamics that they themselves cited as problematic.

Comparison of the clinical and pathologic staging in patients undergoing radical cystectomy for bladder cancer.

PURPOSE: Radical cystectomy (RCx) is perhaps the most effective therapeutic approach for patients with muscle-invasive bladder cancer. Unfortunately, clinical staging is imprecise and the degree of understaging remains high. This study retrospectively evaluated patients undergoing RCx with regard to pathologic outcomes and degree of upstaging to better identify features that may lessen clinical understaging. MATERIALS AND METHODS: 141 consecutive patients with urothelial bladder carcinoma who were candidates for RCx with curative intent were retrospectively evaluated. Preoperative clinical and pathological (i.e. TURBT) features were compared to pathological outcomes in the cystectomy specimen. Patients were also evaluated as to whether cystectomy was performed as their primary (n = 91) versus secondary (n = 50) treatment for recurrent/progressive disease. Date of cystectomy ( 5 years prior to study) was also analyzed. RESULTS: Of the 141 patients, 54% were upstaged on operative pathology. The greatest degree of upstaging occurred in those with invasive disease preoperatively (cT2-T3). Twenty-six percent of all patients had node-positive disease, and 75% of cT3 patients were node-positive. Seven of 101 (7%) patients with clinical T2 disease were unresectable at the time of surgery. In the primary (vs. secondary) RCx group, more patients were upstaged (63% vs. 40%), non-organ confined (62% vs. 38%), and LN positive (31% vs. 20%). In the more modern cohort, the degree of upstaging was not improved. CONCLUSIONS: Pathologic findings after RCx often do not correlate with preoperative staging. Over half of patients undergoing cystectomy are upstaged on their operative pathology. An improved understanding of the relative frequency of upstaging in cystectomy patients may have important implications in the decision-making and selection for neoadjuvant and adjuvant therapies for these high-risk populations.

Comparison of the clinical and pathologic staging in patients undergoing radical cystectomy for bladder cancer

PURPOSE: Radical cystectomy (RCx) is perhaps the most effective therapeutic approach for patients with muscle-invasive bladder cancer. Unfortunately, clinical staging is imprecise and the degree of understaging remains high. This study retrospectively evaluated patients undergoing RCx with regard to pathologic outcomes and degree of upstaging to better identify features that may lessen clinical understaging. MATERIALS AND METHODS: 141 consecutive patients with urothelial bladder carcinoma who were candidates for RCx with curative intent were retrospectively evaluated. Preoperative clinical and pathological (i.e. TURBT) features were compared to pathological outcomes in the cystectomy specimen. Patients were also evaluated as to whether cystectomy was performed as their primary (n = 91) versus secondary (n = 50) treatment for recurrent/progressive disease. Date of cystectomy (< 5 years vs. > 5 years prior to study) was also analyzed. RESULTS: Of the 141 patients, 54 percent were upstaged on operative pathology. The greatest degree of upstaging occurred in those with invasive disease preoperatively (cT2-T3). Twenty-six percent of all patients had node-positive disease, and 75 percent of cT3 patients were node-positive. Seven of 101 (7 percent) patients with clinical T2 disease were unresectable at the time of surgery. In the primary (vs. secondary) RCx group, more patients were upstaged (63 percent vs. 40 percent), non-organ confined (62 percent vs. 38 percent), and LN positive (31 percent vs. 20 percent). In the more modern cohort, the degree of upstaging was not improved. CONCLUSIONS: Pathologic findings after RCx often do not correlate with preoperative staging. Over half of patients undergoing cystectomy are upstaged on their operative pathology. An improved understanding of the relative frequency of upstaging in cystectomy patients may have important implications in the decision-making and selection for neoadjuvant and adjuvant therapies ...

Initial incision of lateral pelvic fascia and early ligation of vascular pedicles during radical prostatectomy: potential to reduce positive margin rates.

OBJECTIVE: To report on our experience with a recently published technique to reduce positive margin (PM) rates (involving early incision of the lateral pelvic fascia, early release of the prostate and Denonvilliers' fascia off the rectum), with the additional modification of early ligation of the lateral vascular pedicles during radical retropubic prostatectomy (RP), as reducing PM rates continues to be an important oncological goal in RP. PATIENTS AND METHODS: One hundred consecutive men (mean age 61 years, pretreatment prostate-specific antigen level 8.9 ng/mL, and estimated blood loss 502 mL) underwent RP by one surgeon. The initial dissection involves early incision of the lateral pelvic fascia and developing the plane between the prostate and underlying rectum, before any apical dissection. This incision can be made medial to the neurovascular bundles in a nerve-sparing procedure. After this plane is developed, the lateral vascular pedicles to the prostate are also divided. Once these same manoeuvres are used contralaterally, the prostate is lifted off of the rectum and held in place only by the apex and bladder neck. The apical/urethral dissection is then carried out conventionally, followed by dissection/transection of the seminal vesicles and the bladder neck. RESULTS: The pathological stage included T2a (11%), T2b (69%), T3a (17%), T3b (3%), and N+ (2%); 20 patients had capsular penetration, at the posterolateral (in 15) and anterior aspect (in five) of the gland. The PM rate for the 100 consecutive patients was 13%, with PMs at the apex in 10, the base in two and posterolateral gland in one. No patient had a PM at the site of capsular penetration. When patients were stratified by low-moderate risk (pT2 and Gleason sum < or = 7) vs high risk (pT3 or Gleason sum > 7), the PM rates were 7.9% and 29.2%, respectively. CONCLUSIONS: Initial dissection of the lateral pelvic fascia, including developing a "perirectal pocket", and early ligation of the lateral pedicles, resulted in a low PM rate during RP. This experience supports the previous observations that early development of the pre-rectal fat plane may allow for more precise dissection below all layers of Denonvilliers' fascia and with a wider margin of periprostatic tissue.

Laboratory evaluation of the patient with erectile dysfunction.

Erectile dysfunction (ED) as a clinical entity is a problem that more than 50% of men will face as they age, and it can adversely affect overall quality of life and impact sexual partners. Our understanding of the pathophysiology and the multiple risk factors that contribute to ED has led to successful treatments, both non- surgical and surgical, over the past two decades. Now, more nonurologists and primary care providers are seeing patients for their initial evaluation. It is imperative that they approach the workup and evaluation in a logical and efficient manner. After a thorough history and physical examination, followed by a relevant and systematic laboratory evaluation, most cases can be effectively identified and medical treatment can be initiated. However, patients who continue to have erectile difficulties or fail initial oral treatment are candidates for specialized evaluation of vascular, neurogenic, and hormonal dysfunction, which can lead to more specific treatment or possible surgical management.

Urinary tract infections in women.

Urinary tract infections in female patients are exceedingly common. One third of all women with an initial UTI demonstrate recurrence, and one third of those recurrences are in the first 6 months. The potential morbidity of RUTIs is high, especially in infants, the elderly, and pregnant patients. Uropathogenic bacteria reside in the rectal vault, colonize the vagina, and then ascend per the urethra into the bladder. These bacteria avidly adhere to uroepithelial cells using pili and can further ascend to the kidneys. Empiric treatment of an initial uncomplicated UTI is acceptable, but recurrences warrant a urine culture and possible radiographic evaluation. Fluoroquinolones are now first-line medications for UTIs, except in geographic areas where resistance to TMP-SMX is still low. Response to therapy should govern the need for additional treatment. Certain motivated patients with three or more UTIs per year should be considered for prophylaxis therapy.