Androgen receptor variant-7 regulation by tenascin-c induced src activation.

BACKGROUND: Bone metastatic prostate cancer does not completely respond to androgen-targeted therapy and generally evolves into lethal castration resistant prostate cancer (CRPC). Expression of AR-V7- a constitutively active, ligand independent splice variant of AR is one of the critical resistant mechanisms regulating metastatic CRPC. TNC is an extracellular matrix glycoprotein, crucial for prostate cancer progression, and associated with prostate cancer bone metastases. In this study, we investigated the mechanisms that regulate AR-V7 expression in prostate cancer cells interacting with osteogenic microenvironment including TNC. METHODS: Prostate cancer/preosteoblast heterotypical organoids were evaluated via immunofluorescence imaging and gene expression analysis using RT-qPCR to assess cellular compartmentalization, TNC localization, and to investigate regulation of AR-V7 in prostate cancer cells by preosteoblasts and hormone or antiandrogen action. Prostate cancer cells cultured on TNC were assessed using RT-qPCR, Western blotting, cycloheximide chase assay, and immunofluorescence imaging to evaluate (1) regulation of AR-V7, and (2) signaling pathways activated by TNC. Identified signaling pathway induced by TNC was targeted using siRNA and a small molecular inhibitor to investigate the role of TNC-induced signaling activation in regulation of AR-V7. Both AR-V7- and TNC-induced signaling effectors were targeted using siRNA, and TNC expression assessed to evaluate potential feedback regulation. RESULTS: Utilizing heterotypical organoids, we show that TNC is an integral component of prostate cancer interaction with preosteoblasts. Interaction with preosteoblasts upregulated both TNC and AR-V7 expression in prostate cancer cells which was suppressed by testosterone but elevated by antiandrogen enzalutamide. Interestingly, the results demonstrate that TNC-induced Src activation regulated AR-V7 expression, post-translational stability, and nuclear localization in prostate cancer cells. Treatment with TNC neutralizing antibody, Src knockdown, and inhibition of Src kinase activity repressed AR-V7 transcript and protein. Reciprocally, both activated Src and AR-V7 were observed to upregulate autocrine TNC gene expression in prostate cancer cells. CONCLUSION: Overall, the findings reveal that prostate cancer cell interactions with the cellular and ECM components in the osteogenic microenvironment plays critical role in regulating AR-V7 associated with metastatic CRPC. Video Abstract.

Silicosis.

Occupational exposures to dust with elevated levels of respirable crystalline silica in artificial stone increase workers' risk for silicosis.

Experimental Evolution of Campylobacter jejuni Leads to Loss of Motility, rpoN (σ54) Deletion and Genome Reduction.

Evolution experiments in the laboratory have focused heavily on model organisms, often to the exclusion of clinically relevant pathogens. The foodborne bacterial pathogen Campylobacter jejuni belongs to a genus whose genomes are small compared to those of its closest genomic relative, the free-living genus Sulfurospirillum, suggesting genome reduction during the course of evolution to host association. In an in vitro experiment, C. jejuni serially passaged in rich medium in the laboratory exhibited loss of flagellar motility-an essential function for host colonization. At early time points the motility defect was often reversible, but after 35 days of serial culture, motility was irreversibly lost in most cells in 5 independently evolved populations. Population re-sequencing revealed disruptive mutations to genes in the flagellar transcriptional cascade, rpoN (σ54)-therefore disrupting the expression of the genes σ54 regulates-coupled with deletion of rpoN in all evolved lines. Additional mutations were detected in virulence-related loci. In separate in vivo experiments, we demonstrate that a phase variable (reversible) motility mutant carrying an adenine deletion within a homopolymeric tract resulting in truncation of the flagellar biosynthesis gene fliR was deficient for colonization in a C57BL/6 IL-10-/- mouse disease model. Re-insertion of an adenine residue partially restored motility and ability to colonize mice. Thus, a pathogenic C. jejuni strain was rapidly attenuated by experimental laboratory evolution and demonstrated genomic instability during this evolutionary process. The changes observed suggest C. jejuni is able to evolve in a novel environment through genome reduction as well as transition, transversion, and slip-strand mutations.

Pelvic Ring Fracture During a Professional Surfing Event at the Banzai Pipeline.

We present a case of a 32-y-old male professional surfer who sustained an isolated pelvic ring fracture after wiping out on a large wave and striking the ocean floor during a world championship tour surfing competition in Hawaii. The surfer was rescued by the water patrol lifeguards, evaluated by onsite medical staff, and stabilized for transfer and subsequent surgical management. As surfing and surfing competitions become increasingly popular, medical staff and event organizers must be aware of the possibility for severe, life-threatening injuries during surfing events. Although infrequent, staff must be prepared to manage these injuries, including immediate resuscitation, stabilization, analgesia, and transfer to definitive care. We hope this case encourages not only surf event organizers and medical staff, but also staff of all professional and recreational water sports, to increase their preparedness to stabilize and treat both life-threatening and minor injuries. Expeditious and appropriate treatment of an injured athlete has the potential to decrease morbidity and mortality while maximizing the athletes' functional outcome after injury.

An Osteopathic Approach to Chronic Pain Management.

Chronic pain reduces quality of life and productivity, costing billions in health care dollars and lost revenue. Physicians routinely prescribe opioids, which has led to opioid addiction and overdose. The US surgeon general recommends nonpharmacologic treatment for patients with chronic pain. A paradigm shift is necessary for patients to partner with physicians to take control of their own health. This article outlines the cognitive behavioral approaches, nonopioid therapies, and nonpharmacologic therapies that osteopathic physicians can integrate in their treatment of patients with chronic pain.

A comparative study of ChIP-seq sequencing library preparation methods.

BACKGROUND: ChIP-seq is the primary technique used to investigate genome-wide protein-DNA interactions. As part of this procedure, immunoprecipitated DNA must undergo "library preparation" to enable subsequent high-throughput sequencing. To facilitate the analysis of biopsy samples and rare cell populations, there has been a recent proliferation of methods allowing sequencing library preparation from low-input DNA amounts. However, little information exists on the relative merits, performance, comparability and biases inherent to these procedures. Notably, recently developed single-cell ChIP procedures employing microfluidics must also employ library preparation reagents to allow downstream sequencing. RESULTS: In this study, seven methods designed for low-input DNA/ChIP-seq sample preparation (Accel-NGS® 2S, Bowman-method, HTML-PCR, SeqPlex™, DNA SMART™, TELP and ThruPLEX®) were performed on five replicates of 1 ng and 0.1 ng input H3K4me3 ChIP material, and compared to a "gold standard" reference PCR-free dataset. The performance of each method was examined for the prevalence of unmappable reads, amplification-derived duplicate reads, reproducibility, and for the sensitivity and specificity of peak calling. CONCLUSIONS: We identified consistent high performance in a subset of the tested reagents, which should aid researchers in choosing the most appropriate reagents for their studies. Furthermore, we expect this work to drive future advances by identifying and encouraging use of the most promising methods and reagents. The results may also aid judgements on how comparable are existing datasets that have been prepared with different sample library preparation reagents.

Treatment outcomes after combination interventional and cognitive motivational counseling on analgesic medication use in patients with chronic spine pain.

BACKGROUND: Pain interventionists can interrupt pain through anesthetic blockade of neural transmission to virtually any part of the body. Temporary pain relief can be achieved by the direct application of targeted anesthetic. Diagnostically, nerve blocks help identify specific pain generators, refine differential diagnosis, and disrupt the neural transmission mechanisms to stop pain generation peripherally. OBJECTIVE: This study of patients with chronic spine pain was conducted to test the hypothesis that decreasing pain through interventional techniques coupled with cognitive motivational counseling can be highly effective in reducing chronic pain interference, reliance on prescription opioids, and enhancing overall function and quality of life. STUDY DESIGN: Retrospective case series. SETTING: Rehabilitation center. PATIENTS: This study involved a retrospective cohort of 78 consecutive patients with spine pain that underwent interventional procedures and cognitive motivational counseling, as well as a comparison group of 77 consecutive patients that underwent interventional procedures only. OUTCOME MEASURES: Pain intensity (DoD VAS), Functional capacity (DoD SS), Global Appraisal (PGIC), Pain site measurement (Drawing), and prescription medication use questionnaires were administered at initial evaluation and after treatment. Pre- and post-treatment changes were compared using paired t-tests. Chi-squared analysis was performed pre- and post-treatment for medication use. RESULTS: The pre- and post-treatment scores for pain intensity, function, and global appraisal demonstrated significant response to treatment (P < 0.001) for the combined interventional and cognitive motivational group (P < 0.001) and the interventional only group (P < 0.05). Compared to initial intake, opioid (P < 0.01), benzodiazepine (P < 0.01), muscle relaxant (P < 0.05), and antidepressant/antianxiolytic (P < 0.05) use only decreased for the combined interventional and cognitive motivational group. LIMITATIONS: This is a retrospective study using medical records and patient self-reported symptoms with possible missed coding and no true random selection, assignment, or genuine control group comparison. CONCLUSION: This study's results support the hypothesis that a combined interventional and cognitive motivational counseling treatment program can be effective in decreasing spine pain, reducing prescription pain medication use, and improving overall quality of life in chronic spine pain patients.

Draft genome sequences of two Campylobacter jejuni clinical isolates, NW and D2600.

The Campylobacter jejuni human clinical isolates NW and D2600 colonized C57BL/6 interleukin-10-deficient (IL-10(-/-)) mice without inducing a robust inflammatory response (J. A. Bell et al., BMC Microbiol. 9:57, 2009). We announce draft genome sequences of NW and D2600 to facilitate comparisons with strains that induce gastrointestinal inflammation in this mouse model.

Touch--more than a basic science.

The potency of touch in osteopathic manipulative treatment (OMT) is physically realized within the musculoskeletal, immune, nervous, and endocrine systems. Psychologically, touch supports a verbal and tactile interaction that is both diagnostic and therapeutic. Touch is a 2-way street that adds meaning and depth to the patient-physician experience. The relationship between touching and being touched offers a potentially powerful and intense deepening of the patient-physician relationship that emerges within the palpatory examination and treatment. Empathic communication, through word or deed, allows a therapeutic, synchronized healing to occur. In the present article, the authors provide a rationale to sensitize and invigorate osteopathic physicians to routinely evaluate and treat patients using their skillful touch.

Standing genetic variation in contingency loci drives the rapid adaptation of Campylobacter jejuni to a novel host.

The genome of the food-borne pathogen Campylobacter jejuni contains multiple highly mutable sites, or contingency loci. It has been suggested that standing variation at these loci is a mechanism for rapid adaptation to a novel environment, but this phenomenon has not been shown experimentally. In previous work we showed that the virulence of C. jejuni NCTC11168 increased after serial passage through a C57BL/6 IL-10(-/-) mouse model of campylobacteriosis. Here we sought to determine the genetic basis of this adaptation during passage. Re-sequencing of the 1.64 Mb genome to 200-500 X coverage allowed us to define variation in 23 contingency loci to an unprecedented depth both before and after in vivo adaptation. Mutations in the mouse-adapted C. jejuni were largely restricted to the homopolymeric tracts of thirteen contingency loci. These changes cause significant alterations in open reading frames of genes in surface structure biosynthesis loci and in genes with only putative functions. Several loci with open reading frame changes also had altered transcript abundance. The increase in specific phases of contingency loci during in vivo passage of C. jejuni, coupled with the observed virulence increase and the lack of other types of genetic changes, is the first experimental evidence that these variable regions play a significant role in C. jejuni adaptation and virulence in a novel host.

Celebrex offers a small protection from root resorption associated with orthodontic movement.

UNLABELLED: Tooth movement results from alveolar bone resorption/deposition following application of orthodontic forces, and root resorption can be an undesirable complication associated with this process. No treatment for external root resorption is available to date. OBJECTIVE: To determine if COX-2 inhibitors like Celebrex are effective in protecting root resorption associated with orthodontic forces. METHODS: A force of 80 grams was applied to the left maxillary first molars of 7-week-old female Wistar rats using nickel titanium closed coil springs attached to the cervical area of the incisors with 0.010 stainless-steel ligature wires. Twenty animals were divided into three experimental groups: one receiving no treatment, the second receiving 25mg/kg, and the third receiving 50 mg/kg of celecoxib (Celebrex) in their drinking water. Rats were maintained on a soft diet and euthanized two weeks after initial placement of the force. Paraffin-embedded sections of the right (control) and left (experimental) maxillae were stained with H&E and the areas of root resorption were examined by counting the number of lacunaes in the roots. RESULTS: No difference in the distance of tooth movement (0.5 mm/two weeks) was seen in all three groups. The rats that received the low dose of Celebrex showed no statistically significant difference in root resorption than that of the rats that received no dose. The rats that received the high dose of Celebrex showed a lower number of lacunaes (mean = 3.5) than that of the control group (mean 10.2; p=0.02). CONCLUSIONS: Administration of Celebrex during the application of orthodontic forces does not interfere with tooth movement and appears to offer some slight protection against root resorption.

Efficacy of venlafaxine for the long term treatment of chronic pain with associated major depressive disorder.

BACKGROUND: This was an open-label, single-center study of the long-term efficacy and effectiveness of venlafaxine extended release (XR) in the treatment of chronic pain and depression in outpatients. All patients have been diagnosed with major depressive disorder (MDD) of various types, with or without chronic pain, and had previously failed treatment with either tricyclic antidepressants (TCAs) or selective serotonin reuptake inhibitors (SSRIs). METHODS: Efficacy of treatment was determined using the 21-item Hamilton Rating Scale for Depression (HAMD-21), the Visual Analogue Scale (VAS) for the evaluation of pain, and a 12-item quality of life scale (QOL). Patients were treated in an unblended open trial for 1 year with 150 mg or more of venlafaxine XR once daily. RESULTS: After 1 year of treatment, 21-item Hamilton Rating Scale for Depression, Visual Analogue Scale, and quality of life scores were significantly improved from permanent baseline scores. CONCLUSION: These data show long-term efficacy and effectiveness of venlafaxine XR, a serotonin (5-HT) and norepinephrine (NE) and dopamine (DA) reuptake inhibitor antidepressant agent, having analgesic properties.