Multicenter Retrospective Analysis of Intradiscal Condoliase Injection Therapy for Lumbar Disc Herniation.

Background and Objectives: Intradiscal injection of Condoliase (chondroitin sulfate ABC endolyase), a glycosaminoglycan-degrading enzyme, is employed as a minimally invasive treatment for lumbar disc herniation (LDH) and represents a promising option between conservative treatment and surgical intervention. Since its 2018 approval in Japan, multiple single-site trails have highlighted its effectiveness, however, the effect of LDH types, and influences of patient age, sex, etc., on treatment success remains unclear. Moreover, data on teenagers and elderly patients has not been reported. In this retrospective multi-center study, we sought to classify prognostic factors for successful condoliase treatment for LDH and assess its effect on patients < 20 and ≥70 years old. Materials and Methods: We reviewed the records of 137 LDH patients treated through condoliase at four Japanese institutions and assessed its effectiveness among different age categories on alleviation of visual analog scale (VAS) of leg pain, low back pain and numbness, as well as ODI and JOA scores. Moreover, we divided them into either a "group-A" category if a ≥50% improvement in baseline leg pain VAS was observed or "group-N" if VAS leg pain improved <50%. Next, we assessed the differences in clinical and demographic distribution between group-A and group-N. Results: Fifty-five patients were classified as group-A (77.5%) and 16 patients were allocated to group-N (22.5%). A significant difference in Pfirrmann classification was found between both cohorts, with grade IV suggested to be most receptive. A posterior disc angle > 5° was also found to approach statical significance. In all age groups, average VAS scores showed improvement. However, 75% of adolescent patients showed deterioration in Pfirrmann classification following treatment. Conclusions: Intradiscal condoliase injection is an effective treatment for LDH, even in patients with large vertebral translation and posterior disc angles, regardless of age. However, since condoliase imposes a risk of progressing disc degeneration, its indication for younger patients remains controversial.

Development of DS-5573a: A novel afucosylated mAb directed at B7-H3 with potent antitumor activity.

B7-H3 is highly overexpressed in a variety of human clinical tumors, and its expression is significantly associated with poor outcomes. In our study, we aimed to develop new antitumor mAbs by employing cancer cell immunization, and succeeded in generating a mouse anti-human B7-H3 antibody (M30) that shows antitumor activity. M30 was humanized (Hu-M30), and an afucosylated Hu-M30 (DS-5573a) was also generated. To assess the potency of DS-5573a as a therapeutic mAb, we characterized this mAb and evaluated its antitumor activity in vitro and in vivo. Flow cytometry analysis showed that B7-H3 proteins were expressed on various types of cancer cell lines broadly, and DS-5573a binds to IgC1 and IgC2 domains of human B7-H3. Antibody-dependent cellular cytotoxicity activity of DS-5573a was drastically enhanced against medium to high B7-H3-expressing cancer cell lines MDA-MB-231 and NCI-H322. DS-5573a also induced high antibody-dependent cellular cytotoxicity activity against low B7-H3-expressing cancer cell line COLO205, whereas Hu-M30 induced little activity against it. In addition, DS-5573a was found to be a novel anti-B7-H3 antibody which showed antibody-dependent cellular phagocytosis activity. Furthermore, DS-5573a showed dose-dependent and significant antitumor efficacy (0.03-3 mg/kg) in MDA-MB-231-bearing SCID mice (which have functional natural killer cells and macrophages), but little antitumor efficacy in NOG mice (which lack natural killer cells and have reduced macrophage function). These results suggest that antitumor activity of DS-5573a is mediated by effector cells, and this mAb could be a promising antitumor therapy for patients with a wide range of B7-H3-expressing tumors.

Neural cell adhesion molecule 2 as a target molecule for prostate and breast cancer gene therapy.

In adenovirus-derived gene therapy, one of the problems is the difficulty in specific targeting. We have recently demonstrated that monoclonal antibody (mAb) libraries screened by fiber-modified adenovirus vector (Adv-FZ33), which is capable of binding to immunoglobulin-G (IgG), provide a powerful approach for the identification of suitable target antigens for prostate cancer therapy. Hybridoma libraries from mice immunized with androgen-dependent prostate cancer cell line LNCaP were screened and mAb were selected. Through this screening, we obtained one mAb, designated LNI-29, that recognizes a glycoprotein with an apparent molecular mass of 100 kD. It was identified as neural cell adhesion molecule 2 (NCAM2). Some prostate and breast cancer cell lines highly expressed NCAM2 whereas normal prostate cell lines expressed NCAM2 at low levels. In contrast to the low efficiency of gene transduction by Adv-FZ33 with a control antibody, LNI-29-mediated Adv-FZ33 infection induces high rates of gene delivery in NCAM2-positive cancers. NCAM2-mediated therapeutic gene transduction of uracil phosphoribosyltransferase (UPRT) had a highly effective cytotoxic effect on NCAM2-positive cancer cells, whereas it had less of an effect in cases with a control antibody. In conclusion, NCAM2 should be a novel gene therapy target for the treatment of prostate and breast cancer.

Datopotamab Deruxtecan, a Novel TROP2-directed Antibody-drug Conjugate, Demonstrates Potent Antitumor Activity by Efficient Drug Delivery to Tumor Cells.

Trophoblast cell surface antigen 2 (TROP2) is highly expressed on various epithelial tumors and correlates with poor prognosis. We developed the novel TROP2-directed antibody-drug conjugate (ADC), datopotamab deruxtecan (Dato-DXd, DS-1062a), with a potent DNA topoisomerase I inhibitor (DXd), and evaluated its antitumor activity and safety profiles in preclinical models.The pharmacologic activity and mechanism of action of Dato-DXd were investigated in several human cancer cell lines and xenograft mouse models including patient-derived xenograft (PDX) models. Safety profiles were also assessed in rats and cynomolgus monkeys.Dato-DXd bound specifically to TROP2 and was internalized into tumor cells followed by intracellular trafficking to lysosome and DXd release, which induced DNA damage and apoptosis in TROP2-expressing tumor cells in vitro. Dato-DXd exhibited in vivo antitumor activity with DNA damage induced by the accumulated DXd in TROP2-expressing xenograft tumors, but neither isotype control IgG-ADC nor anti-TROP2 antibody had this effect. Dato-DXd also showed potent antitumor activity with tumor regression in several TROP2-expressing xenograft tumors including NSCLC PDX models. Safety profiles of Dato-DXd in rats and cynomolgus monkeys were acceptable.Dato-DXd demonstrated potent antitumor activity against TROP2-expressing tumors by efficient payload delivery into tumors and acceptable safety profiles in preclinical models. These results suggest Dato-DXd could be a valuable treatment option for patients with TROP2-expressing tumors in the clinical setting.

Health Beneficial Food Emulsifier Produced from Fishery Byproducts.

The bioavailability of DHA-bound phospholipids, especially the DHA-bound lysophospholipid (DHA-LPL) could be considered the most effective DHA chemical forms for DHA accretion in the brain. Such a DHA-LPL should also have very high emulsifying stability performance based on its analogy with conventional soy LPL. Therefore, in this study, we describe two fishery byproducts, rich in DHA-bound phospholipids, to derive DHA-LPL via sn-1 positional specific lipase partial hydrolysis of the phospholipids. Through this reaction, the DHA composition increased to 43.8 % from 29.1 % in the salmon head phospholipid-derived DHA-LPL, and to 84.0 % from 47.4 % in the squid meal phospholipid-derived DHA-LPL. In fact, these obtained DHA-LPLs exhibited far higher emulsifying stability than the conventional food emulsifiers in the market. For example, the prepared high-purity squid meal phospholipid-derived LPL sustained an emulsion form for a week even under 80°C. Thus, food emulsifiers produced from fishery byproducts are considered to exhibit very high values of both in a sense of outstandingly high health benefits and sustaining emulsions even under very high temperatures.

Doxorubicin-loaded calcium phosphate cement in the management of bone and soft tissue tumors.

Several materials have been used as drug delivery systems to maintain a high concentration of anticancer drugs at localized sites. The feasibility of using doxorubicin-loaded calcium phosphate cement (CPC) as a new material, which can release the drug as well as fill a postoperative bony defect, was investigated. The mechanical strength of cylinders of doxorubicin-loaded CPC was not lower than that of CPC alone. Culture medium incubated with doxorubicin-loaded CPC from 1 to 7 days suppressed the proliferation of RMT-1 E4 rat breast cancer cells. In rabbits with doxorubicin-loaded CPC in their femurs, histological examinations showed diffuse edematous changes in the medullary space, but neither fracture nor skin necrosis occurred. Doxorubicin-loaded CPC markedly inhibited the proliferation of sarcoma 180 cells in the mouse air-pouch model. These results indicate that doxorubicin-loaded CPC may be useful in the local treatment of malignant bone and soft tissue tumors.

Electricity generation from rice bran in microbial fuel cells.

BACKGROUND: Rice bran is a by-product of the rice milling process and mostly discarded in Japan. Although many studies have shown that microbial fuel cells (MFCs) are able to generate electricity from organic wastes, limited studies have examined MFCs for generating electricity from rice bran. FINDINGS: Laboratory-scale single-chamber MFCs were inoculated with paddy field soil and supplied with rice bran for examining electricity generation. Power outputs and microbiome compositions were compared between MFCs containing pure water as the liquid phase (MFC-W) and those containing mineral solution (MFC-M). Polarization analyses showed that both MFCs successfully generated electricity with the maximum power densities of 360 and 520 mW m-2 (based on the projected area of anode) for MFC-W and MFC-M, respectively. Amplicon-sequencing analyses revealed that Trichococcus and Geobacter specifically occurred in anode biofilms in MFC-W and MFC-M, respectively. CONCLUSIONS: The results suggest that rice bran is a feasible fuel by itself for generating electricity in MFCs.

Interaction of the parathyroid hormone receptor with the 14-3-3 protein.

The receptor for parathyroid hormone (PTH) and PTH-related protein (PTHrP) regulates calcium homeostasis, bone remodeling and skeletal development. 14-3-3 proteins bind to signaling proteins and act as molecular scaffolds and regulators of subcellular localization. We show that the parathyroid hormone receptor (PTHR) interacts with 14-3-3 and the proteins colocalize within the cell. 14-3-3 interacts with the C-terminal tail of the receptor containing a consensus 14-3-3 binding motif, but additional binding sites are also used. Protein kinase-A treatment of the receptor and especially the C-terminal tail reduces 14-3-3 binding. The expressed C-terminal tail is primarily localized in the nucleus, supporting the function of a putative nuclear localization signal that could be involved in the previously described nuclear localization of PTHR. The observed interaction between PTHR and the 14-3-3 protein implies that 14-3-3 could contribute to regulation of PTHR signaling.

Upregulation of SR-PSOX/CXCL16 and recruitment of CD8+ T cells in cardiac valves during inflammatory valvular heart disease.

OBJECTIVE: SR-PSOX/CXCL16 is a transmembrane chemokine and is implicated in activated CD8+ T cell trafficking. In the present study, we examined the expression pattern of SR-PSOX/CXCL16 in the heart and investigated a potential role of SR-PSOX/CXCL16 in inflammatory valvular heart disease. METHODS AND RESULTS: Initial expression of SR-PSOX/CXCL16 in murine embryos was detected in endothelial cells lining endocardial cushions in the forming heart at E11.5. From mid-gestation to adult, expression of this gene in the heart was exclusively observed in valvular endothelial cells. Examination of SR-PSOX/CXCL16 expression in human cardiac valves demonstrated that SR-PSOX/CXCL16 was strongly expressed in valvular and neocapillary endothelial cells in patients with infective endocarditis. SR-PSOX/CXCL16 expression in neocapillary endothelial cells was also observed in patients with rheumatic and atherosclerotic valvular disease. Moreover, CD8+ T cells were distributed closely to endothelial cells expressing SR-PSOX/CXCL16. In vitro adhesion assays showed that SR-PSOX/CXCL16 induced adhesion of activated CD8+ T cells to vascular cell adhesion molecule-1 (VCAM-1) through very late antigen-4 (VLA-4) activation. Furthermore, SR-PSOX/CXCL16 stimulated interferon-gamma (IFN-gamma) production by CD8+ T cells. CONCLUSIONS: SR-PSOX/CXCL16 may be involved in CD8+ T cell recruitment through VLA-4 activation and stimulation of IFN-gamma production by CD8+ T cells during inflammatory valvular heart disease.

Cell surface-anchored SR-PSOX/CXC chemokine ligand 16 mediates firm adhesion of CXC chemokine receptor 6-expressing cells.

Direct contacts between dendritic cells (DCs) and T cells or natural killer T (NKT) cells play important roles in primary and secondary immune responses. SR-PSOX/CXC chemokine ligand 16 (CXCL16), which is selectively expressed on DCs and macrophages, is a scavenger receptor for oxidized low-density lipoprotein and also the chemokine ligand for a G protein-coupled receptor CXC chemokine receptor 6 (CXCR6), expressed on activated T cells and NKT cells. SR-PSOX/CXCL16 is the second transmembrane-type chemokine with a chemokine domain fused to a mucin-like stalk, a structure very similar to that of fractalkine (FNK). Here, we demonstrate that SR-PSOX/CXCL16 functions as a cell adhesion molecule for cells expressing CXCR6 in the same manner that FNK functions as a cell adhesion molecule for cells expressing CX(3)C chemokine receptor 1 (CX(3)CR1) without requiring CX(3)CR1-mediated signal transduction or integrin activation. The chemokine domain of SR-PSOX/CXCL16 mediated the adhesion of CXCR6-expressing cells, which was not impaired by treatment with pertussis toxin, a Galphai protein blocker, which inhibited chemotaxis of CXCR6-expressing cells induced by SR-PSOX/CXCL16. Furthermore, the adhesion activity was up-regulated by treatment of SR-PSOX/CXCL16-expressing cells with a metalloprotease inhibitor, which increased surface expression levels of SR-PSOX/CXCL16. Thus, SR-PSOX/CXCL16 is a unique molecule that not only attracts T cells and NKT cells toward DCs but also supports their firm adhesion to DCs.

Effect of retinoic acid on murine preosteoblastic MC3T3-E1 cells.

Retinoic acid (RA) plays an important role in bone metabolism in vivo through osteoclast activation and bone resorption. Retinoid X-activated receptor beta (RXRbeta) has been implicated in the genetic spinal defect of ossification of the posterior longitudinal ligament (OPLL). In this study, we examined the effects of 9-cis RA and all-trans RA (ATRA) on the proliferation, differentiation, and RXRbeta expression of the murine preosteoblastic cell line MC3T3-E1. Both 9-cis RA and ATRA dose-dependently inhibited the increase in total soluble protein content at concentrations of 10 and 100 nM after 4 and 8 d co-culture with MC3T3-E1 cells. The inhibitory effect of 9-cis RA was slightly stronger than that of ATRA. Histone H4 mRNA expression was dose-dependently suppressed by both RAs on day 1. Alkaline phosphatase activity was increased by both RAs at 10 and 100 nM concentrations on day 4, with 9-cis RA-induced activity slightly stronger than that of ATRA. Osteopontin mRNA expression was increased by both RAs on day 1, but was suppressed on day 4. Bone Gla protein mRNA expression was inhibited by 10 and 100 nM 9-cis RA and by 100 nM ATRA on day 14. RXRbeta mRNA expression was increased by 9-cis RA, an RXRbeta ligand, in a dose-dependent manner. Our results suggested that while both RAs suppressed proliferation and stimulated the maturation of preosteoblastic MC3T3-E1 cells, 9-cis RA was slightly more potent than ATRA. It also appeared that RAs may contribute to the development of heterotopic ossification, including OPLL.

The effect of magnetic stimulation on unloaded soleus muscle of rat: changes in myosin heavy chain mRNA isoforms.

This study assessed the potential application and the effectiveness of functional magnetic stimulation (FMS) for preventing skeletal muscle atrophy in adult rats. FMS using magnetic stimulator was performed to rat soleus muscle by placing a round magnetic coil on the back of 3rd-5th lumbar vertebral level at 20 Hz frequency for 60 min/day up to 10 days. A reverse transcriptase-polymerase chain reaction was applied to evaluate relative amounts of mRNAs specific to four myosin heavy chain (MHC) isoforms [MHCIbeta, MHCIIa, MHCIIb, and MHCIId(x)] in rat soleus muscle during contractile activity by magnetic stimulation. Ten-day unloading by hindlimb suspension induced a drastic decrease of MHCIbeta and MHCIIa mRNA expressions, while MHCIIb and MHCIId(x) mRNA was not decreased. The magnetic stimulation resuscitated the down-regulation of the mRNA levels of MHCIbeta and MHCIIa. These results suggest that magnetic stimulation on acute atrophied muscles is useful for preventing the muscle atrophy.

Osteosarcoma occurring in osteogenesis imperfecta.

We describe a case history of a 24-year-old male with osteogenesis imperfecta (OI) who developed osteosarcoma of the left thigh. High-dose ifosfamide therapy caused marked tumor regression of multiple lung metastases. Immunohistochemically, the tumor cells were diffusely positive for the p53 protein. Mutation of the p53 gene was not detected by direct genomic sequencing of exons 4-8. The radiographic characteristics, including irregularly distributed osteolytic lesions and cortical discontinuity, should not be confused with hyperplastic callus formation, a benign process. A biopsy is critical to establish the differential diagnosis between osteosarcoma and common hyperplastic callus formation in OI; however, it must be applied with great care.

Congenital pseudoarthrosis of the tibia treated with low-intensity pulsed ultrasound stimulation (LIPUS).

A 16-year-old Japanese boy was admitted to our institution in September 2000 because no apparent callus had appeared around a fracture after 6 weeks of cast fixation. Physical examination revealed a tenderness of the right lower leg, and multiple small subcutaneous tumors and café-au-lait spots in extremities and trunk. Radiographs showed the fracture of the right lower tibia with bony sclerosis and a localized fusiform osteolytic lesion at the fracture site. The affected tibia bowed anteriorly and the medullary space in the lower tibia was narrow. A diagnosis of Boyd type IV congenital pseudoarthrosis of the tibia was made. Treatment with low-intensity pulsed ultrasound (US) stimulation (LIPUS) was administered for 20 min/day, and a nonweight-bearing gait was continued with a cast or brace fixation. At 6 months after the treatment, a small amount of bridging callus was seen. We continued the treatment for 1 year until the solid fusion was observed on radiographs and the patient started full-weight-bearing.

Protective effects of HFE7A, mouse anti-human/mouse Fas monoclonal antibody against acute and lethal hepatic injury induced by Jo2.

HFE7A is a mouse anti-human/mouse Fas monoclonal antibody which, protects mice from fulminant hepatitis induced by Jo2. Herein, we report on the mechanism of the protective effect of HFE7A against Jo2-induced acute and lethal hepatic injury. HFE7A reduced the serum aminotransferase level which was elevated after Jo2 injection. HFE7A also inhibited caspase activation and mitochondrial depolarization in hepatocytes derived from apoptosis induced by Jo2 injection. The protective effect of HFE7A against Jo2-induced apoptosis in mouse hepatocytes was reproducible in vitro. The cell death and caspase activation in isolated mouse hepatocytes were induced by incubating these cells with Jo2 in vitro, and HFE7A inhibited the cell death and caspase activation in mouse hepatocytes in a dose-dependent manner. The affinity of HFE7A to mouse Fas was lower than that of Jo2. The binding of Jo2 to neither recombinant mouse Fas nor mouse hepatocytes was inhibited by an excessive amount of HFE7A. Interestingly, HFE7A bound to hepatocytes isolated from Fas knockout mice. From these results, it is suggested that HFE7A may exert a protective effect against Jo2-induced hepatitis not by competitively inhibiting the binding of Jo2 to Fas on hepatocytes, and that a distinct molecule other than Fas may possibly be involved in the protective effect of HFE7A against Jo2-induced hepatic injury.

Pigmented villonodular synovitis of the elbow treated with the Tsuge wide joint exposure technique.

Pigmented villonodular synovitis (PVNS) is a benign, locally aggressive disease of the synovium; its cause remains unclear. The most frequently involved joint is the knee, followed by the hip, ankle, wrist, and shoulder. Pigmented villonodular synovitis of the elbow joint is rare. Synovectomy is currently believed to be the best treatment for PVNS. Open or arthroscopic synovectomy is usually selected. During synovectomy for PVNS, the possibility of local recurrence after surgery must be considered. The recurrence rate after synovectomy of any joint for PVNS is approximately 40%. Therefore, surgical treatment for PVNS of the elbow requires sufficient removal of the lesion. For good functional results, prevention of postoperative joint stiffness is also necessary. This article describes a case of a 29-year-old woman with PVNS of the right elbow who was treated by total synovectomy using the Tsuge technique. Tsuge reported a new surgical technique for debridement arthroplasty using a posterolateral approach to the elbow in 1987. He has also reported using this procedure during arthroplasty for posttraumatic stiff elbow and for synovectomy in rheumatoid arthritis. This approach permits easy dislocation of the elbow and provides a good view of the whole joint. Although the recurrence rate of PVNS of the elbow is high, our patient has retained good elbow function with no evidence of local recurrence at 30 months postoperatively.

Osteosarcoma of the proximal fibula. An analysis of 13 cases in the northern Japan.

Osteosarcoma is the most common form of malignant bone tumor that occurs during childhood and adolescence. The proximal fibula is a relatively rare site for osteosarcoma. We reviewed 305 cases of osteosarcoma registered at the Tohoku Musculoskeletal Tumor Society (TMTS) between 1975 and 1999. Thirteen patients (4.3%) had their osteosarcomas localized in the proximal fibula. Conventional fibroblastic osteosarcoma accounted for 46% of the cases in this series. Limb-sparing surgery was performed in all 13 patients during initial surgery, and the peroneal nerve was preserved in 4 cases. These 4 cases developed local recurrences, but additional wide excision or radiation had a beneficial effect on the recurrences. In our series, the patients showed a 5-year survival rate 76 per cent. The postoperative function of the knee remained good despite various reattachment procedures of lateral co-lateral ligament. As well as resection of the proximal fibula, our results indicate that osteosarcoma of the proximal fibula has a good prognosis for cases who undergo adequate initial surgery.

Discal cyst associated with myxoid change and apoptosis of herniated disc materials: a case report.

Discal cyst is a lumbar intraspinal cyst communicating with intervertebral disc, and previously reported series described the wall of these cysts as consisting of dense fibrous connective tissue. We report a 29-year-old Japanese male with discal cyst showing unusual histological features. Clinical symptoms in the current case as well as imaging features including discography were similar to those previously reported.However, the wall of the cyst consisted of disc material with myxoid degeneration. In addition, apoptosis of chondrocytes was diffusely observed in the herniated disc material. The current case was considered a histological variant of discal cyst. Myxoid degeneration of herniated disc material with diffuse apoptotic change of chondrocytes was probably associated with the formation of discal cyst.