Association of Preoperative Osteoporotic Vertebral Compression Fractures with Muscle Atrophy in Lumbar Spinal Stenosis: A Retrospective Cohort Study.

Objectives: Osteoporotic vertebral compression fractures (OVCFs) are common in older individuals and lead to pain, spinal deformities, and limited mobility. Paraspinal muscle function correlates with fracture severity, and this association may be more significant in patients with lumbar spinal stenosis (LSS). However, studies on the effects of OVCFs are lacking. This study aimed to investigate the relationship between OVCFs, fat infiltration, and muscle atrophy in patients with LSS. Methods: This study included 177 patients with preoperative LSS, of whom 16 had OVCFs and 161 did not. Lumbar lordosis angle, fat infiltration, and paraspinal muscle atrophy were evaluated in these patients. Information on patient characteristics such as smoking, diabetes, hemodialysis, steroid use, American Society of Anesthesiologists score, and bladder or bowel dysfunction were obtained from medical records. Logistic regression analysis was conducted to identify factors independently associated with OVCF. Results: Patients in the OVCF group were significantly older (P=0.006) than those without fractures, and a higher proportion of the OVCF group showed muscle atrophy (P=0.034). Significant variables and those with moderate effect sizes were included in the logistic regression analysis. Muscle atrophy (P=0.028) was independently associated with OVCF. Conclusions: Muscle atrophy was associated with preoperative OVCFs in patients with LSS. Identifying OVCFs in these patients may underscore the importance of tailored treatment and rehabilitation strategies for the paraspinal muscles.

Effectiveness of an educational approach between adolescents and adults with patellofemoral pain syndrome: a scoping review with meta-analysis.

[Purpose] This study aimed to compare the effectiveness of an educational approach between adolescents and adults with patellofemoral pain syndrome. [Participants and Methods] The PROSPERO protocol number is CRD42022362680. Four databases (PubMed, CINAHL, Scopus, Cochrane) were searched. Randomized control trials on the effectiveness of the educational approach for patellofemoral pain syndrome were compared between adolescents and adults. Two investigators independently screened eligible studies. Two randomized control trials proceeded through meta-analysis with a visual analogue scale in the mid-term (from 18 weeks to 6 months). [Results] The meta-analysis results indicated that no significant difference was observed between education and combined education with exercise. A tendency to improve in the intervention of combined education with exercise for adolescents was noted, whereas no effect of education for adults was observed. [Conclusion] Treatment strategies combining education with exercise for patellofemoral pain syndrome may differ between adolescents and adults.

Immediate Effects of Foam Roller and Stretching to the Lead Hip on Golfers Swing: A Randomized Crossover Trial.

Golfers with decreased range of motion (ROM) of their leading hip internal rotation (IR) have increased lumbar rotation ROM and load. This study investigated the effects of foam roller (FR) applied to their leading hip muscles combined with stretching to the leading hip together with lumbar rotation ROM during the golf swing. The study design was a crossover design. Subjects were allocated to one of two groups comprising FR and dynamic stretching (FR + DS) or practice swing. Motion analysis was used to evaluate hip and lumbar angles during the golf swing. Data were compared using analysis of variance with Bonferroni correction using paired t-test's post hoc. The association between lead hip IR angle and lumbar spine left rotation (Lrot) angle was investigated using correlation analysis. Lead hip IR ROM during the golf swing was significantly greater in the FR + DS group (p = 0.034). The FR + DS group showed a moderate negative correlation between lead hip IR ROM and lower lumbar spine Lrot ROM during the golf swing (r = -0.522). The application of FR + DS might be useful to increase lead hip IR angle during the golf swing. Moreover, the application of FR + DS improves lead hip IR angle and may decrease lumbar spine rotation.

Effects of cross-training on motor function and length of stay after total hip arthroplasty: A randomized controlled trial.

BACKGROUND: There is no consensus about which training methods will give better early outcomes after total hip arthroplasty (THA). OBJECTIVE: To investigate the short-term effects of cross trainer exercise on physical function and walking ability following THA. METHODS: Fifty patients who underwent THA were randomly allocated into two groups. The intervention program was started 3 days after surgery. The main physical function results were pain, hip range of motion, knee extensor strength, single-leg stance time, and walking performance test. In addition, the number of days of requiring to walk and the length of hospital stay were recorded. RESULTS: In the comparison between groups at discharge, the patients in the cross trainer group had significantly less hip pain while walking, improvement in knee extensor strength, increased single-leg stance time, as well as increased walking speed and stride length at discharge. The number of days required to walk and length of stay were also significantly lower in the intervention group. CONCLUSIONS: Conclusions: Cross trainer exercise commencing 3 days postoperatively improves physical function and walking ability after THA.

Mechanisms of EGFR-TKI-Induced Apoptosis and Strategies Targeting Apoptosis in EGFR-Mutated Non-Small Cell Lung Cancer.

Homeostasis is achieved by balancing cell survival and death. In cancer cells, especially those carrying driver mutations, the processes and signals that promote apoptosis are inhibited, facilitating the survival and proliferation of these dysregulated cells. Apoptosis induction is an important mechanism underlying the therapeutic efficacy of epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs) for EGFR-mutated non-small cell lung cancer (NSCLC). However, the mechanisms by which EGFR-TKIs induce apoptosis have not been fully elucidated. A deeper understanding of the apoptotic pathways induced by EGFR-TKIs is essential for the developing novel strategies to overcome resistance to EGFR-TKIs or to enhance the initial efficacy through therapeutic synergistic combinations. Recently, therapeutic strategies targeting apoptosis have been developed for cancer. Here, we review the state of knowledge on EGFR-TKI-induced apoptotic pathways and discuss the therapeutic strategies for enhancing EGFR-TKI efficiency. We highlight the great progress achieved with third-generation EGFR-TKIs. In particular, combination therapies of EGFR-TKIs with anti-vascular endothelial growth factor/receptor inhibitors or chemotherapy have emerged as promising therapeutic strategies for patients with EGFR-mutated NSCLC. Nevertheless, further breakthroughs are needed to yield an appropriate standard care for patients with EGFR-mutated NSCLC, which requires gaining a deeper understanding of cancer cell dynamics in response to EGFR-TKIs.

Inter-examiner reliability in identifying lumbar paraspinal muscle atrophy by lumbar paraspinal muscle atrophy index, a novel parameter.

[Purpose] To evaluate the inter-examiner reliability of our novel parameter, the lumbar paraspinal muscle atrophy index, in identifying the lumbar paravertebral muscle atrophy. [Participants and Methods] The study group consisted of 225 adults, with a mean age of 64.7 (range, 21-89) years, who underwent posterior lumbar spinal surgery for degenerative spinal disease at our hospital between July 2013 and June 2017. Preoperative axial T2-weighted magnetic resonance images were used to evaluate the lumbar paraspinal muscle atrophy index and observe the presence or absence of severe lumbar paraspinal muscle atrophy. The lumbar paraspinal muscle atrophy index was calculated at each intervertebral level, from L1-2 through L4-5, once by two examiners, and the Cohen's kappa statistic was used to calculate the inter-examiner agreement of the classification of the presence or absence of atrophy at each level. [Results] The agreement was high (kappa, 0.79-0.88) for the lumbar paraspinal muscle atrophy index at all levels, except at the L3-4 level (kappa, 0.49). The lower kappa statistic at L3-4 likely reflects the unique morphological characteristics at this level. [Conclusion] The lumbar paraspinal muscle atrophy index is a new, simple, easy-to-use, and sufficiently reliable parameter to identify lumbar paraspinal atrophy.

Effects of a Groin Pain Prevention Program in Male High School Soccer Players: A Cluster-Randomized Controlled Trial.

Background: Groin pain is frequently reported by soccer players. A prevention program incorporating the Copenhagen adduction exercise (CAE) has been shown effective in decreasing adductor muscle injury in semi-professional soccer players. However, the effect of such programs on groin pain in high school soccer players is unknown. No reports have examined the effects of combining the CAE with other targeted exercises such as the Nordic hamstrings exercise (NHE). Purpose: To evaluate the preventative effects of exercise on groin pain in high school soccer players in three groups comprising NHE alone, combined NHE and CAE, and a control group. Design: Randomized Controlled Clinical Trial. Methods: A cluster randomized controlled trial spanning 16 weeks was conducted on 202 high school soccer players from seven high schools in a Japanese under 18 Soccer League. Players were allocated to either group A (three schools, 66 players) receiving the CAE alone, or group B (two schools, 73 players) receiving the CAE and NHE, or control group C (two schools, 63 players) who performed their usual training. This study compared the number of injuries, injury rate, and severity of the injury of groin pain in these three groups. Results: The number of players injured was less in group B (4 players) compared to C (18 players), and time-lost to soccer was less in groups A (6 players) and B (3 players) compared to group C (16 players). Injury rates were significantly lower in groups A and B, with the relative risk of injury compared to group C of 0.42 (95% CI 0.19% to 0.90%) in group A and 0.19 (95% CI 0.07 to 0.54) in group B. Conclusion: A 16-week program incorporating the CAE in training sessions in high school soccer players reduced the incidence of groin pain and which may be related to injury severity according to time -lost to soccer, however the combination of both the CAE and NHE may be more effective than the CAE alone. Level of Evidence: 2b.

Diverse Mechanisms of Resistance against Osimertinib, a Third-Generation EGFR-TKI, in Lung Adenocarcinoma Cells with an EGFR-Activating Mutation.

Osimertinib, a third-generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), is used as a first-line treatment for patients with EGFR-mutant non-small cell lung cancer (NSCLC). However, the mechanisms underlying its anticancer activity, particularly the subsequent development of acquired resistance, are unclear. Herein, we investigated the mechanisms underlying the development of osimertinib resistance by treating NSCLC PC-9 cells (harboring an EGFR-activating mutation) with osimertinib, thereby developing five resistant cell lines, i.e., AZDR3, AZDR6, AZDR9, AZDR11, and AZDR14. The amplification of wild-type EGFR in AZDR3 cells and wild-type EGFR and KRAS in AZDR6 cells was also studied. AZDR3 cells showed dependence on EGFR signaling, in addition to afatinib sensitivity. AZDR9 cells harboring KRASG13D showed sensitivity to MEK inhibitors. Furthermore, combination treatment with EGFR and IGF1R inhibitors resulted in attenuated cell proliferation and enhanced apoptosis. In AZDR11 cells, increased Bim expression could not induce apoptosis, but Bid cleavage was found to be essential for the same. A SHP2/T507K mutation was also identified in AZDR14 cells, and, when associated with GAB1, SHP2 could activate ERK1/2, whereas a SHP2 inhibitor, TNO155, disrupted this association, thereby inhibiting GAB1 activation. Thus, diverse osimertinib resistance mechanisms were identified, providing insights for developing novel therapeutic strategies for NSCLC.

Effects of incorporating elliptical trainer exercise during rehabilitation on physical function and self-reported outcomes after total hip arthroplasty: a randomized controlled trial.

[Purpose] To investigate the effects of incorporating elliptical trainer exercise in early rehabilitation after total hip arthroplasty on physical function and self-reported outcomes. [Participants and Methods] Participants with independent gait prior to total hip arthroplasty underwent conventional postoperative physiotherapy and were divided into two groups. The intervention group additionally underwent elliptical trainer exercise, while the control group underwent a walking program. The main outcomes were low back and hip region pain, lower limb muscle strength, single-leg stance time, Timed Up & Go Test results, 10 m walking test results, hip disability and osteoarthritis outcome score, and modified fall efficacy scale score. These outcomes were evaluated preoperatively, at discharge, and at 1 and 3 months postoperatively. [Results] Fifty participants (including 40 females; age, 68.3 ± 10.8 years) participated in this study. Physical function evaluations showed a significant improvement in hip region pain during walking at discharge. Knee extensor strength, single-leg stance time, stride length, and walking speed were significantly greater in the intervention group at discharge and at 1 and 3 months postoperatively. The modified fall efficacy scale score significantly improved in the intervention group 1 month postoperatively. [Conclusion] Elliptical trainer exercise and conventional physiotherapy in the early postoperative period contribute to improved physical function and walking ability and improvement in the fear of falling.

Determining a preventive strategy for ankle sprain injury through a questionnaire survey of coaches of junior high school basketball teams.

[Purpose] The purpose of this study was to determine a preventive strategy for ankle sprain injury through a questionnaire survey of coaches of junior high school basketball teams. [Participants and Methods] The questionnaire comprised questions concerning coaches' characteristics, prevention-related content, and collaboration with medical staff. We distributed the questionnaire to 108 coaches, of which 48 (41.7%) responded. [Results] The results revealed that none of the participant coaches had ever collaborated with medical staff. [Conclusion] Additional effort from medical staff is needed to hold seminars and contact coaches of junior high school teams in order to conduct an injury prevention program.

Risk Factor Analysis for Fat Infiltration in the Lumbar Paraspinal Muscles in Patients With Lumbar Degenerative Diseases.

INTRODUCTION: This study aimed to investigate factors related to fat infiltration in patients with lumbar degenerative diseases (lumbar disc herniation and/or spinal stenosis), examining a wide range of potential risk variables. MATERIALS AND METHODS: We studied consecutive adult patients who underwent posterior lumbar spinal surgery for degenerative diseases at our hospital between July 2013 and June 2017. Preoperative magnetic resonance imaging was used to evaluate the presence or absence of fat infiltration at the level of the L4-5 lumbar paraspinal muscles using Kjaer's evaluation method. Patients without fat infiltration (0%-10%) were rated as grade 0, grade 1 for moderate fat infiltration (10%-50%), and grade 2 for severe fat infiltration (>50%). Patients were then divided into two groups: Group A (without fat infiltration, grade 0) and Group B (with fat infiltration, grade 1 or 2). Detailed patient clinical data were collected and analyzed. RESULTS: A total of 205 consecutive patients were enrolled; 54 (26.3%) patients were assigned to Group A and 151 (73.7%) to Group B. Logistic regression analysis revealed two independent risk factors for fat infiltration of the lumbar paraspinal muscles: female sex and older age (P < .001). DISCUSSION: Fat infiltration of the lumbar paraspinal muscles is reported to be associated with the development of pain and dysfunction of the lumbar region and postoperative complications of spinal instrumented fusion surgery. To the best of our knowledge, no previous studies have identified female sex and older age as independent risk factors for fat infiltration in the lumbar paraspinal muscles using multivariate analysis. CONCLUSIONS: Female sex and older age were independent risk factors for fat infiltration in the lumbar paraspinal muscles. The results of the current study may provide useful information for the study of preventive measures for fat infiltration.

Factors affecting early knee-flexion range of motion after total knee arthroplasty.

[Purpose] To investigate the factors affecting the knee-flexion range of motion in the early period after total knee arthroplasty. [Participants and Methods] Ninety-nine patients who had undergone total knee arthroplasty at our hospital between 2016 and 2019 were allocated into two groups based on the presence of a 110° knee-flexion range of motion at 14 days post-surgery. From medical records, we extracted data for the participants' basic attributes and preoperative/postoperative physical function (knee-flexion range of motion, Timed Up & Go Test results, resting/walking pain according to a numerical rating scale, and knee-extension muscle strength). Postoperative physical function was measured 14 days post-surgery. [Results] Preoperative knee-flexion range of motion, preoperative femorotibial angle, postoperative knee-extensor strength, and postoperative Timed Up & Go Test value differed significantly as factors related to achieving a 110° knee-flexion range of motion. Through further statistical analyses, we selected the preoperative knee-flexion range of motion, preoperative femorotibial angle, preoperative Timed Up & Go Test result, and postoperative knee-extension strength as factors affecting the knee-flexion range of motion at 14 days post-surgery. [Conclusion] Preoperative knee-flexion range of motion, preoperative femorotibial angle, preoperative Timed Up & Go Test result, and postoperative knee-extension strength influence knee-flexion range of motion at 14 days after total knee arthroplasty, and our findings indicate the effectiveness of active physiotherapy interventions.

Inter-rater reliability between two examiners with different professional roles in the evaluation of fat infiltration in the lumbar paraspinal muscles using magnetic resonance imaging.

[Purpose] To clarify the inter-rater reliability of the evaluation criteria for paraspinal muscle fat infiltration on magnetic resonance images between two examiners with different professional roles in interdisciplinary physical therapy teams. [Participants and Methods] In this retrospective study, we reviewed the clinical data of 225 patients with degenerative lumbar diseases who underwent posterior lumbar surgery at our hospital. A physical therapist and a spinal surgeon visually quantified fat infiltration of the multifidus muscles at the level of L4/5 on the preoperative magnetic resonance images of the patients using Kjaer's criteria (Grade 0: 0-10%, Grade 1: 10-50%, and Grade 2: >50%). We used the kappa coefficient to assess inter-rater reliability. [Results] The participants included 142 males and 83 females (mean age, 64.7 years; range, 21-89 years). The number of patients with grades 0/1/2 were 50/160/15, respectively, for examiner 1; and 59/155/11, respectively, for examiner 2. The kappa coefficient was 0.69, indicating a substantial agreement. [Conclusion] Our study, which is the first to assess the inter-rater reliability of Kjaer's criteria between examiners with different medical occupations, revealed that these criteria could be a reliable tool for evaluating fat infiltration in the multifidus muscles and sharing information between interdisciplinary physical therapy teams.

Effects of functional movement screen training in high-school baseball players: A randomized controlled clinical trial.

BACKGROUND: In recent years, the functional movement screen (FMS) and FMS training have attracted attention as a means of preventing injury, but no studies have examined the effect of such training in high-school baseball players. The aim of this study was to clarify the effect of FMS training on FMS score, physical function and baseball performance in high-school baseball players. METHODS: Subjects in this randomized controlled clinical trial were high-school male baseball players assigned to either an FMS training group (intervention group) or a control group. The intervention group performed FMS training 4 times per week for 12 weeks. FMS ability, physical function, and baseball performance were measured prior to the intervention, 8, 12, and 24 weeks after the intervention in the subjects' school environment. RESULTS: A total of 71 baseball players aged 15 to 17 years were recruited and assigned to either an intervention group (n = 37) or control group (n = 34). There was no significant difference in the characteristics of participants between the 2 groups. Most FMS scores improved to 12 weeks after continued training. In the intervention group compared with the control group, deep squat, hurdle step, inline lunge, active straight leg raise, trunk stability push-up and rotary stability FMS score, total FMS score and eyes closed single leg stance time significantly increased after 8 weeks of training. While hurdle step, inline lunge, active straight leg raise, trunk stability push-up, total FMS score, and eyes closed single leg stance time significantly increased, pitching ball speed significantly decreased at the end of the 12 week training period. Eyes closed single leg stance time and feeling of fatigue significantly improved 12 weeks after training. The number of subjects who scored less than 14 for the total FMS score in the intervention group compared with control group were significantly less after 8 and 12 weeks of FMS training. CONCLUSION: FMS training for 8 weeks contributes to improving FMS scores for high-school baseball players, but FMS scores go down if FMS training is not continued. TRIAL REGISTRATION: University Hospital Medical Information Network Center, Tokyo, Japan: UMIN000027553. Registered on May 30, 2017.

A Novel, Circulating Tumor Cell Enrichment Method Reduces ARv7 False Positivity in Patients with Castration-Resistant Prostate Cancer.

BACKGROUND: The AR-V7 splice variant is a cause of castration-resistant prostate cancer (CRPC). However, testing for the presence of AR-V7 by real-time polymerase chain reaction (RT-PCR) shows AR-V7 positivity in healthy individuals. We hypothesized that the positivity reflects contamination by hematopoietic cells. We tried a novel circulating tumor cell (CTC) enrichment instrument, using Celsee, to clear hematopoietic cells. METHODS: We tested whole blood or Celsee-enriched samples for AR-V7 by RT-PCR, and included samples from 41 CRPC patients undergoing sequential therapy. We evaluated the associations between AR-V7 status and clinical factors. We evaluated factors affecting AR-V7 positivity. RESULTS: AR-V7 positivity was lower in Celsee-enriched than in whole blood specimens. AR-V7 and clinical factors did not predict the therapy effectiveness. We found no significant differences in the effectiveness of enzalutamide/abiraterone (Enz/Abi) upon AR-V7 evaluation. All AR-V7 positive patients had resistance to Enz/Abi. Docetaxel (DTX), cabazitaxel (CBZ), and Radium223 treatment showed no significant difference in the treatment effectiveness, regardless of AR-V7 presence. AR-V7 was more frequently positive than Extent of disease (EOD) 2 in cases with bone metastases. CONCLUSION: Celsee CTC enrichment suppresses AR-V7 false positivity. All AR-V7 positive patients presented resistance to Enz/Abi. DTX, CBZ, and Radium223 were effective and remain treatment options. AR-V7 positivity should progressively appear in patients with advanced bone metastases.

Effects of Nordic Hamstring Exercise on Hamstring Injuries in High School Soccer Players: A Randomized Controlled Trial.

We evaluated a range of physical characteristics related to hamstring injuries, as well as the Nordic Hamstring Exercise compliance rate, and whether this influenced the rate hamstring injury. Subjects comprised 259 male soccer players from seven high schools randomly clustered into two groups, a Nordic Hamstring Exercise group and a control group. Training and match time were logged, as well as details of hamstring injury, and subsequent time lost to hamstring injury recorded over a period of 27 weeks. The Nordic Hamstring Exercise compliance rate, injury rate per 10000 playing hours and time-lost-to-sport-injury rate were calculated. The relative risk and hamstring injury severity were also calculated. The hamstring injury rate was 1.04/10 000 h in the control group and 0.88/10 000 h in the intervention group. The relative risk for hamstring injury was 1.14. The time-lost to injury rate was 1116.3/10 000 h in the control group and 113.7/10 000 h in the intervention group; with relative risk 9.81. The Nordic Hamstring Exercise in high school soccer players significantly reduced hamstring injury severity compared to a control intervention. Our results indicate that the time-lost to injury rate should be taken into account when analyzing the severity of hamstring injury.

Development of a Highly Sensitive Technique for Capturing Renal Cell Cancer Circulating Tumor Cells.

PURPOSE: Liquid biopsy is becoming increasingly important as a guide for selecting new drugs and determining their efficacy. In urological cancer, serum markers for renal cell and urothelial cancers has made the development of liquid biopsy for these cancers strongly desirable. Liquid biopsy is less invasive than conventional tissue biopsy is, enabling frequent biopsies and, therefore, is considered effective for monitoring the treatment course. Circulating tumor cells (CTCs) are a representative liquid biopsy specimen. In the present study, we focused on developing our novel technology for capturing renal cell cancer (RCC)-CTCs using an anti-G250 antibody combined with new devices. Basic experiments of our technology showed that it was possible to detect RCC-CTC with a fairly high accuracy of about 95%. Also, RCC-CTC was identified in the peripheral blood of actual RCC patients. Additionally, during the treatment course of the RCC patient, change in the number of RCC-CTC was confirmed in one case. We believe that the technology we developed will be useful for determining the treatment efficacy and drug selection for the treatment of renal cell cancer (RCC). In order to solve issues such as thresholds setting of this technology, large-scale clinical trials are expected.

Three-Dimensional Motion Analysis of the 2nd Cervical Spinous Process at End Range Cervical Rotation in Different Scapular Positions Using 3D Digitizer.

OBJECTIVE: The study used a 3D digitizer to determine three-dimensional motion analysis of the 2nd cervical (C2) spinous process at end range cervical rotation with the scapula in different positions. METHODS: 30 healthy adults participated in this study. Different scapula positions were adopted bilaterally and positioned passively at normal resting, depression, adduction, and abduction. Under each scapula position, bilateral end range cervical rotation and displacement of the C2 spinous process were analyzed by a 3D digitizer. RESULTS: Displacement of the C2 spinous process relative to the occiput was significantly correlated with range of cervical rotation under all scapular positions (p<0.05). However, there were no significant differences between end range cervical rotation and displacement of the C2 spinous process relative to the occiput in any scapular position. CONCLUSION: These results suggest that measurement of upper cervical mobility using the 3D digitizer is a reliable method that holds promise in the evaluation of people with cervical spine disorders.

Enhancement of IL-2-induced cytotoxicity by interferon-alpha in renal cell carcinoma.

Metastatic renal cell carcinoma (mRCC) treatment consists of molecular targeted agents and cytokines that have fundamentally different mechanisms of action. Clinical responses also differ; complete response is rare with molecular targeted agents but is sometimes achieved with cytokine therapies. Because of the relatively high efficacy of combination therapy with low-dose interleukin-2 (IL-2) and interferon-alpha (IFN-alpha) against mRCC, it is important to reevaluate cytokine therapies in vitro. Here, we show that when IL-2 is administered in combination with IFN-alpha, a stronger cytotoxic effect of PBMCs on RCC cell lines is observed than when IL-2 is administered alone. The upregulation of TNF-related apoptosis-inducing ligand on NK cell by IL-2 and suppression of regulatory T cells (Tregs) by IFN-alpha were recognized at the same time when cytotoxicity of peripheral blood mononuclear cells (PBMCs) was enhanced. IL-2 is known to activate natural killer cell cytotoxicity; however, IL-2 also stimulates Treg expansion, which enhances immunosuppression. On the other hand, IFN-alpha negatively regulates Treg cells, thereby increasing the function of immune effector cells. Our in vitro results may explain, at least in part, the clinical efficacy of combination low-dose IL-2 and IFN-alpha therapy against mRCC.

Induction of matrix metalloproteinase gene expression in an endothelial cell line by direct interaction with malignant cells.

Mouse endothelial TKD2 cells in monolayers were cocultured with various human cell lines for 24 h, and the expression of several secreted matrix metalloproteinases (MMP) and cell adhesion molecules was examined by real-time reverse transcription-polymerase chain reaction using mouse-specific primers. Coculture with normal fibroblasts did not elicit the expression of these molecules, but coculture with cancer cells induced the expression of MMP-3, MMP-9 and MMP-10 mRNA in endothelial cells, and in normal mouse embryonic fibroblasts. The induction of MMP mRNA was dependent on direct cell adhesion, as separate culture of A549 cells in Boyden chambers did not induce MMP mRNA, and neutralizing antibody against VLA-4 abolished the induction. An inhibitor of phosphatidylinositol-3-phosphate kinase strongly suppressed the induction of MMP-3, MMP-9 and MMP-10 mRNA, and expression of the dominant-negative mutant of phosphatidylinositol-3-phosphate kinase also decreased the induction. It was suggested that intracellular reactive oxygen species (ROS) levels were increased in TKD2 cells following adhesion to cancer cells. ROS scavengers decreased the levels of MMP induction, and roterone, an inhibitor of mitochondrial complex I, strongly suppressed the induction of MMP-3, MMP-9 and MMP-10. The depletion of mitochondria in TKD2 cells decreased the induction of MMP-9, but the induction of MMP-3 and MMP-10 was not affected. These results indicate that the adhesion of cancer cells to endothelial cells activates several distinct signaling pathways to induce MMP gene expression, and the pathways for MMP-3, MMP-9 and MMP-10 are partly different. For the induction of MMP-9, mitochondria participate in induction, possibly through the production of ROS.