Self-Renewal Inhibition in Breast Cancer Stem Cells: Moonlight Role of PEDF in Breast Cancer.

Breast cancer is the leading cause of death among females in developed countries. Although the implementation of screening tests and the development of new therapies have increased the probability of remission, relapse rates remain high. Numerous studies have indicated the connection between cancer-initiating cells and slow cellular cycle cells, identified by their capacity to retain long labeling (LT+). In this study, we perform new assays showing how stem cell self-renewal modulating proteins, such as PEDF, can modify the properties, percentage of biomarker-expressing cells, and carcinogenicity of cancer stem cells. The PEDF signaling pathway could be a useful tool for controlling cancer stem cells' self-renewal and therefore control patient relapse, as PEDF enhances resistance in breast cancer patient cells' in vitro culture. We have designed a peptide consisting of the C-terminal part of this protein, which acts by blocking endogenous PEDF in cell culture assays. We demonstrate that it is possible to interfere with the self-renewal capacity of cancer stem cells, induce anoikis in vivo, and reduce resistance against docetaxel treatment in cancer patient cells in in vitro culture. We have also demonstrated that this modified PEDF protein produces a significant decrease in the percentage of expressed cancer stem cell markers.

Cadaveric and Ultrasound Validation of Percutaneous Electrolysis Approach at the Achilles Tendon as a Potential Treatment for Achilles Tendinopathy: A Pilot Study.

Achilles tendon tendinopathy (AT) is a musculoskeletal condition characterized by pain in the Achilles tendon and impaired physical performance or sport activities. AT is difficult to treat, and the results are variable. Preliminary evidence suggests a positive effect for pain of percutaneous electrolysis in patients with tendinopathy. Our aim was to determine the validity and safety of a percutaneous electrolysis approach targeting the interphase between the Achilles tendon and the Kager's fat with ultrasound imaging in both healthy individuals and on a fresh cadaver model (not ultrasound guiding). A needle was inserted from the medial to the lateral side under the body of the Achilles tendon, just between the tendon and the Kager's triangle, about 5 cm from the insertion of tendon in the calcaneus in 10 healthy volunteers (ultrasound study) and 10 fresh cadaver legs. An accurate needle penetration of the interphase was observed in 100% of the approaches, in both human and cadaveric models. No neurovascular bundle of the sural nerve was pierced in any insertion. The distance from the tip of the needle to the sural nerve was 5.28 ± 0.7 mms in the cadavers and 4.95 ± 0.68 mms in the volunteer subjects, measured in both cases at a distance of 5 cm from the insertion of the Achilles tendon. The results of the current study support that percutaneous electrolysis can be safely performed at the Kager's fat-Achilles tendon interphase if it is US guided. In fact, penetration of the sural nerve was not observed in any needle approach when percutaneous needling electrolysis was performed by an experienced clinician. Future studies investigating the clinical effectiveness of the proposed intervention are needed.

Endogenous Pain Modulation in Response to a Single Session of Percutaneous Electrolysis in Healthy Population: A Double-Blinded Randomized Clinical Trial.

The purpose of this double-blinded randomized controlled trial was to investigate whether percutaneous electrolysis (PE) is able to activate endogenous pain modulation and whether its effects are dependent on the dosage of the galvanic current. A total of 54 asymptomatic subjects aged 18-40 years were randomized into three groups, receiving a single ultrasound-guided PE intervention that consisted of a needle insertion on the lateral epicondyle tendon: sham (without electrical current), low-intensity (0.3 mA, 90 s), and high-intensity (three pulses of 3 mA, 3 s). Widespread pressure pain thresholds (PPT), conditioned pain modulation (CPM), and temporal summation (TS) were assessed in the elbow, shoulder, and leg before and immediately after the intervention. Both high and low intensity PE protocols produced an increase in PPT in the shoulder compared to sham (p = 0.031 and p = 0.027). The sham group presented a significant decrease in the CPM (p = 0.006), and this finding was prevented in PE groups (p = 0.043 and p = 0.025). In addition, high-intensity PE decreased TS respect to sham in the elbow (p = 0.047) and both PE groups reduced TS in the leg (p = 0.036 and p = 0.020) without significant differences compared to sham (p = 0.512). Consequently, a single PE intervention modulated pain processing in local and widespread areas, implying an endogenous pain modulation. The pain processing effect was independent of the dosage administrated.

Implication of Different Tumor Biomarkers in Drug Resistance and Invasiveness in Primary and Metastatic Colorectal Cancer Cell Lines.

Protein expression profiles are directly related to the different properties of cells and are conditioned by the cellular niche. As an example, they are the cause of the characteristic cell plasticity, epithelium-mesenchymal transition (EMT), and drug resistance of cancer cells. This article characterizes ten biomarkers related to these features in three human colorectal cancer cell lines: SW-480, SW-620, and DLD-1, evaluated by flow cytometry; and in turn, resistance to oxaliplatin is studied through dose-response trials. The main biomarkers present in the three studied lines correspond to EpCAM, CD-133, and AC-133, with the latter two in low proportions in the DLD-1 line. The biomarker CD166 is present in greater amounts in SW-620 and DLD-1 compared to SW-480. Finally, DLD-1 shows high values of Trop2, which may explain the aggressiveness and resistance of these cells to oxaliplatin treatments, as EpCAM is also highly expressed. Exposure to oxaliplatin slows cell growth but also helps generate resistance to the treatment. In conclusion, the response of the cell lines is variable, due to their genetic variability, which will condition protein expression and cell growth. Further analyses in this area will provide important information for better understanding of patients' cellular response and how to prevent resistance.

Cadaveric and Ultrasound Validation of Percutaneous Electrolysis Approaches at the Arcade of Frohse: A Potential Treatment for Radial Tunnel Syndrome.

Entrapment of the radial nerve at the arcade of Frohse could contribute to symptoms in patients with lateral epicondylalgia or radial tunnel syndrome. Our aim was to determine the validity of applying percutaneous electrolysis, targeting the supinator muscle at the Frohse's arcade with ultrasound imaging and in a Thiel-embalmed cadaver model (not ultrasound-guiding). Percutaneous electrolysis targeting the supinator muscle was conducted in five healthy volunteers (ultrasound study) and three Thiel-embalmed cadaver forearms. Two approaches, one with the forearm supinated and other with the forearm pronated were conducted. The needle was inserted until the tip reached the interphase of both bellies of the supinator muscle. Accurate needle penetration of the supinator muscle was observed in 100% in both US-imaging and cadaveric studies. No neurovascular bundle of the radial-nerve deep branch was pierced in any insertion. The distance from the tip of the needle to the neurovascular bundle was 15.3 ± 0.6 mm with the forearm supinated, and 11.2 ± 0.6 mm with the forearm pronated. The results of the current study support that percutaneous electrolysis can properly target the supinator muscle with either the forearm in supination or in pronation. In fact, penetration of the neurovascular bundle was not observed in any approach when percutaneous needling electrolysis was performed by an experienced clinician.

Effects of Percutaneous Electrolysis on Endogenous Pain Modulation: A Randomized Controlled Trial Study Protocol.

Percutaneous electrolysis consists of the application of a galvanic electrical current throughout an acupuncture needle. It has been previously hypothesized that needling procedures' neurophysiological effects may be related to endogenous pain modulation (EPM). This protocol study describes the design of a double-blind (participant, assessor) randomized controlled trial with the aim to investigate whether percutaneous electrolysis is able to enhance EPM and whether the effect is different between two applications depending on the dosage of the galvanic electrical current. Seventy-two asymptomatic subjects not reporting the presence of pain symptoms the previous 6 months before the study, aged 18-40 years, are randomized into one of four groups: a control group who does not receive any intervention, a needling group who receives a needling intervention without electrical current, a low-intensity percutaneous electrolysis group (0.3 mA × 90 s), and a high-intensity percutaneous electrolysis group (three bouts of 3 mA × 3 s). Needling intervention consists of ultrasound-guided insertion of the needle on the common extensor tendon of the lateral epicondyle. The primary outcome is conditioned pain modulation (CPM), and secondary outcomes include widespread pressure pain sensitivity (pressure pain thresholds (PPT) over the lateral epicondyle, the cervical spine, and the tibialis anterior muscle) and temporal summation (TS). We expected that percutaneous electrolysis would have a greater influence on CPM than an isolated needling procedure and no intervention. In addition, we also postulated that there might be differences in outcome measures depending on the intensity of the electrical current during the percutaneous electrolysis application. This study makes a new contribution to the field of neurophysiological effects of percutaneous electrolysis and needling interventions.

Dry Needling of a Healthy Rat Achilles Tendon Increases Its Gene Expressions: A Pilot Study.

BACKGROUND: Tendon dry needling is a potential treatment for tendinopathies. Several hypotheses have been proposed to explain its underlying mechanisms. No studies (to the best of our knowledge) have investigated changes in gene expression. OBJECTIVE: To investigate histological and gene expression changes after the application of dry needling to the healthy Achilles tendons of rats. METHODS: Six Sprague-Dawley male rats were randomly divided into two groups: no intervention or dry needling. Dry needling consisted of three sessions (once per week) to the Achilles tendon. Molecular expression of several genes involved in tendon repair and remodeling (e.g., Cox2, Mmp2, Mmp9, Col1a1, Col3a1, Vefg, and Scx) was assessed 7 days after the last needling session (day 28) or 28 days after for the no-intervention group. Histological tissue changes were determined with hematoxylin-eosin analyses. RESULTS: The hematoxylin-eosin-stained images revealed no substantial differences in collagen structure or the presence of inflammatory cells between the dry needling and no-intervention groups. A significant increase in the molecular expression of Cox2, Mmp2, Col3a1, and Scx genes was observed in Achilles tendons treated with dry needling when compared with the no-intervention group. CONCLUSION: This animal pilot study found that the application of dry needling to the healthy Achilles tendons of rats is able to increase the expression of genes associated with collagen regeneration and tissue remodeling of the extracellular matrix with no further histological damage to the tendon.

Changes in Gene Expression Associated with Collagen Regeneration and Remodeling of Extracellular Matrix after Percutaneous Electrolysis on Collagenase-Induced Achilles Tendinopathy in an Experimental Animal Model: A Pilot Study.

Percutaneous electrolysis is an emerging intervention proposed for the management of tendinopathies. Tendon pathology is characterized by a significant cell response to injury and gene expression. No study investigating changes in expression of those genes associated with collagen regeneration and remodeling of extracellular matrix has been conducted. The aim of this pilot study was to investigate gene expression changes after the application of percutaneous electrolysis on experimentally induced Achilles tendinopathy with collagenase injection in an animal model. Fifteen Sprague Dawley male rats were randomly divided into three different groups (no treatment vs. percutaneous electrolysis vs. needling). Achilles tendinopathy was experimentally induced with a single bolus of collagenase injection. Interventions consisted of 3 sessions (one per week) of percutaneous electrolysis or just needling. The rats were euthanized, and molecular expression of genes involved in tendon repair and remodeling, e.g., Cox2, Mmp2, Mmp9, Col1a1, Col3a1, Vegf and Scx, was examined at 28 days after injury. Histological tissue changes were determined with hematoxylin-eosin and safranin O analyses. The images of hematoxylin-eosin and Safranin O tissue images revealed that collagenase injection induced histological changes compatible with a tendinopathy. No further histological changes were observed after the application of percutaneous electrolysis or needling. A significant increase in molecular expression of Cox2, Mmp9 and Vegf genes was observed in Achilles tendons treated with percutaneous electrolysis to a greater extent than after just needling. The expression of Mmp2, Col1a1, Col3a1, or Scx genes also increased, but did not reach statistical significance. This animal study demonstrated that percutaneous electrolysis applied on an experimentally induced Achilles tendinopathy model could increase the expression of some genes associated with collagen regeneration and remodeling of extracellular matrix. The observed gene overexpression was higher with percutaneous electrolysis than with just needling.

Immediate effects of bilateral sacroiliac joint manipulation on plantar pressure distribution in asymptomatic participants.

OBJECTIVE: To investigate the immediate effects of manipulation of bilateral sacroiliac joints (SIJs) on the plantar pressure distribution in asymptomatic participants in the standing position. DESIGN: Randomized, controlled, double-blind clinical trial. PARTICIPANTS: Sixty-two asymptomatic men and women (mean age, 20.66±2.56 years) randomly assigned to 2 groups. INTERVENTIONS: The experimental group underwent mobilization without tension of the hips in the supine position and high-velocity, low-amplitude manipulation in the SIJs bilaterally. The control group underwent only mobilization, without tension of the hips in supine position. OUTCOME MEASURES: Pre- and postintervention outcomes measured by an assessor blinded to the treatment allocation of the participants included a baropodometric analysis performed by using a force platform. Baseline between-group differences were examined with a Kolmogorov-Smirnov test. A chi-square test was used for categorical data. Analysis of covariance (ANCOVA) was used to assess differences between groups, with the preintervention value as covariant (95% confidence level). RESULTS: At baseline, no variables significantly differed between groups. Baropodometric analysis showed statistically significant differences in the location of the maximum pressure point in the experimental group (p=0.028). Pre- and postintervention analysis with ANCOVA showed statistically significant differences between both groups in the left hindfoot load percentage (interaction p=0.0259; ANCOVA p=0.0277), right foot load percentage (ANCOVA p=0.0380), and surface of the right forefoot (interaction p=0.0038). There was also a significant effect in the variables that analyze the entire foot (left foot: surface [interaction p=0.0452], percentage of load [ANCOVA p=0.0295]) and between both groups (right foot: weight [interaction p=0.0070; ANCOVA p=0.0296]). CONCLUSIONS: Sacroiliac joint manipulation applied bilaterally in asymptomatic persons resulted in immediate changes in load distribution on plantar support in the standing position. Study limitations and suggestions for future studies are discussed.

Effects of manual therapy on craniofacial pain in patients with chronic rhinosinusitis: a case series.

OBJECTIVE: Chronic rhinosinusitis (CRS) is thought to develop through an inadequate drainage of nasal and sinus secretions and perpetuated by local mechanical and autonomic nervous system factors. Manual therapy may have an effect on these factors providing symptomatic relief of CRS symptoms. The purpose of this prospective case series was to report the results of manual therapy on a set of patients with craniofacial pain and a diagnosis of CRS. METHODS: Fourteen consecutive patients presenting with a primary report of craniofacial pain and a diagnosis CRS completed self-report questionnaires including the Sinonasal Assessment Questionnaire, Rhinosinusitis Task Force, visual analog scale for craniofacial pain, and pressure pain threshold over 4 sinus points on the face. Patients were seen once a week for 7 consecutive weeks and completed all outcome measures at baseline and subsequent weekly sessions. They received manual therapy interventions only on the second, third, and fifth weekly sessions. RESULTS: No significant changes in outcome measures were observed from baseline to 1 week, where no intervention was applied. Significant improvements were observed on all outcome measures (Ps ≤ .015) for pre- and post-first treatment session, as well as from baseline to 7 weeks (Ps < .001). All patients exhibited a significant decrease in craniofacial pain and increased pressure pain thresholds and reported less severity of their symptoms. CONCLUSION: Patients with craniofacial pain and CRS who were treated with manual therapy demonstrated improvements in all outcome measures only after each treatment session. Our results suggest that manual therapy treatment could be considered as an appropriate alternative treatment of CRS.