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.

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.

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.

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.

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.

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.

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.

Anthracyclines, small-molecule inhibitors of hypoxia-inducible factor-1 alpha activation.

Hypoxia-inducible factor-1 (HIF-1) is a central mediator of cellular responses to low oxygen and has recently become an important therapeutic target for solid tumor therapy. To identify small molecule inhibitors of the HIF-1 transcriptional activation, we have established a high through-put assay system using a stable transformant of mammalian cells that express the luciferase reporter gene construct containing a HIF-1 binding site. Using this system, we screened 5000 cultured broths of microorganisms, and we found that cinerubin (1-hydroxy aclacinomycin B) showed a significant inhibition of the reporter activity induced by hypoxic conditions. In addition, we demonstrated that aclarubicin also inhibited the HIF-1 transcriptional activity under hypoxic conditions, but neither doxorubicin nor daunorubicin inhibited it. Consistent with these results, cinerubin and aclarubicin inhibited the hypoxic induction of the vascular endothelial growth factor (VEGF) protein in HepG2 cells, but neither doxorubicin nor daunorubicin affected it. Thus, our results suggested that some anthracyclines are also acting as angiogenesis inhibitors.

Inhibition of angiogenesis and HIF-1alpha activity by antimycin A1.

We identified antimycin A1 as an inhibitor of the hypoxia-response element (HRE) from screening using a reporter under the control of HRE under hypoxic conditions. Antimycin A1 was effective at 20 pg/ml in inhibiting the reporter activity. The expression of vascular endothelial growth factor (VEGF) mRNA during hypoxia was also inhibited by antimycin A1. Angiogenesis induced by implantation of mouse sarcoma-180 cells was significantly inhibited by non-toxic doses of antimycin A1. Hypoxia inducible factor (HIF)-1alpha protein levels were significantly decreased by antimycin A1, but its mRNA level was not affected. Antimycin A1 is known to be an inhibitor of mitochondrial electron transport system, and depletion of mitochondria abolished antimycin A1-effect, at least in part. Inhibitors of proteasome or protein synthesis did not affect the decrease in HIF-1alpha level induced by antimycin A1. These results indicate that antimycin A1 inhibited angiogenesis through decrease in VEGF production caused by inhibition of HIF-1alpha activation.

Specific inhibition of hypoxia-inducible factor (HIF)-1 alpha activation and of vascular endothelial growth factor (VEGF) production by flavonoids.

Screening using a reporter under the control of the hypoxia-response element (HRE) identified several flavonoids and homoisoflavonoids that inhibit the activation of HRE under hypoxic conditions. Among various compounds, isorhamnetin, luteolin, quercetin, and methyl ophiopogonanone B (MOB) were effective at 3 to 9 microg/ml in inhibiting the reporter activity. The expression of vascular endothelial growth factor (VEGF) mRNA during hypoxia was also inhibited by MOB in HepG2 cells, but the effective doses were 10 to 20 microg/ml. MOB caused destabilization of hypoxia-inducible factor (HIF)-1alpha, as revealed by Western blotting, that was dependent on proteasome activity and the tumor suppressor, p53. The tubular formation and migration of human umbilical vein endothelial cells was also inhibited by MOB. MOB is expected to act as an inhibitor of angiogenesis.