Periodic reciprocating motion of a polymer gel on an aqueous phase synchronized with the Belousov-Zhabotinsky reaction.

A self-oscillating gel induced by the Belousov-Zhabotinsky (BZ) reaction was investigated on an aqueous phase. When the Ru-catalyst in the gel was rapidly oxidized, the gel was accelerated in a direction opposite to the side of oxidation. The gel then returned to its original position while the Ru-catalyst in the gel was slowly reduced. To clarify the mechanism of this periodic reciprocation of the gel, the contact angle between a sessile bubble and the gel and the time-variation of the adhesive force of the gel on the aqueous phase were measured. The experimental results suggest that the periodic reciprocation of the gel is driven by the periodic change in the contact angle of the gel induced by the BZ reaction.

Valgus toe deformity of fourth proximal phalanx due to osteochondroma treated with closing wedge osteotomy.

Osteochondroma is the most common benign tumor of all benign and primary bone tumors. It rarely occurs in the proximal phalanx of the lesser toe. The treatment of osteochondroma usually consists of simple resection. However, if other deformities remain, added procedures may be considered. We report a case of a valgus toe deformity of the fourth proximal phalanx due to osteochondroma. The patient was a 21-year-old man who noticed a valgus deformity of his fourth toe over 10 years earlier. He began to experience pain in his fifth toe because of crossover of the fourth toe when wearing formal shoes. Although resection of osteochondroma was performed, the valgus deformity was not sufficiently corrected. Therefore, closing wedge osteotomy of the proximal phalanx was performed at the same time. A good outcome was achieved for this patient.

Checkrein deformity due to extensor hallucis longus hypotrophy treated with extensor digitorum longus tendon transfer.

Checkrein deformity is a relatively rare condition caused by hypotrophy or adhesion of a tendon after a lower leg injury. The occurrence of this condition due to the dysfunction of the extensor hallucis longus (EHL) is extremely rare. Only a few related case reports have been published, and Z-lengthening of the EHL tendon was performed for almost all patients. We report a case of checkrein deformity due to EHL hypotrophy. The patient was involved in a traffic accident 7 years ago. He sustained left tibial and fibular closed diaphyseal fractures and underwent minimally invasive plate osteosynthesis. He continued to have left great toe symptoms characterized by dorsiflexion of the great toe during ankle plantarflexion. The EHL had become an insufficient power source because of considerable hypotrophy. Therefore, a tendon transfer using the extensor digitorum longus to the second toe was performed as a primary treatment.

Continuous year-round isolation of giant viruses from brackish shoreline soils.

Giant viruses, categorized under Nucleocytoviricota, are believed to exist ubiquitously in natural environments. However, comprehensive reports on isolated giant viruses remain scarce, with limited information available on unrecoverable strains, viral proliferation sites, and natural hosts. Previously, the author highlighted Pandoravirus hades, Pandoravirus persephone, and Mimivirus sp. styx, isolated from brackish water soil, as potential hotspots for giant virus multiplication. This study presents findings from nearly a year of monthly sampling within the same brackish water region after isolating the three aforementioned strains. This report details the recurrent isolation of a wide range of giant viruses. Each month, four soil samples were randomly collected from an approximately 5 × 10 m plot, comprising three soil samples and one water sample containing sediment from the riverbed. Acanthamoeba castellanii was used as a host for virus isolation. These efforts consistently yielded at least one viral species per month, culminating in a total of 55 giant virus isolates. The most frequently isolated species was Mimiviridae (24 isolates), followed by Marseilleviridae (23 isolates), Pandoravirus (6 isolates), and singular isolates of Pithovirus and Cedratvirus. Notably, viruses were not consistently isolated from any of the four samples every month, with certain sites yielding no viruses. Cluster analysis based on isolate numbers revealed that soil samples from May and water and sediment samples from January produced the highest number of viral strains. These findings underscore brackish coastal soil as a significant site for isolating numerous giant viruses, highlighting the non-uniform distribution along coastlines.

Effects of Pyrolysis Temperature and Chemical Modification on the Adsorption of Cd and As(V) by Biochar Derived from Pteris vittata.

Phytoremediation can be applied successfully to solve the serious worldwide issue of arsenic (As) and cadmium (Cd) pollution. However, the treatment of biomass containing toxic elements after remediation is a challenge. In this study, we investigated the effective use of biomass resources by converting the As hyperaccumulator P. vittata into biochar to adsorb toxic elements. Plant biomass containing As was calcined at 600, 800, and 1200 °C, and its surface structure and adsorption performances for As(V) and Cd were evaluated. Pyrolysis at 1200 °C increased the specific surface area of the biochar, but it did not significantly affect its adsorption capacity for toxic elements. The calcined biochar had very high adsorption capacities of 90% and 95% for As(V) and Cd, respectively, adsorbing 6000 mmol/g-biochar for As(V) and 4000 mmol/g-biochar for Cd. The As(V) adsorption rate was improved by FeCl3 treatment. However, the adsorption capacity for Cd was not significantly affected by the NaOH treatment. In conclusion, it was found that after phytoremediation using P. vittata biomass, it can be effectively used as an environmental purification material by conversion to biochar. Furthermore, chemical modification with FeCl3 improves the biochar's adsorption performance.

Effect of Asymptomatic Severe Aortic Stenosis on Outcomes of Individuals Aged 80 and Older.

OBJECTIVES: To examine the effect of asymptomatic severe aortic stenosis (AS) on mortality late in life. DESIGN: Retrospective cohort study. SETTING: Large medical center. PARTICIPANTS: Asymptomatic adults aged 80 and older (mean age 86 ± 4; N=1,060, n=569 women) with preserved left ventricular ejection fraction (LVEF; > 50%); 927 (87.5%) with no AS, 70 (6.6%) with nonsevere AS, and 63 (5.9%) with severe AS. MEASUREMENTS: Information was collected on demographic characteristics, comorbidities, and laboratory and echocardiographic data. Survival was assessed according to longest follow-up available. RESULTS: During a mean follow-up of 2.2 ± 2.3 years, there were 203 (19%) deaths, 51 of which were from cardiovascular causes. Four-year estimates of survival were 72% for no AS, 58% for nonsevere AS, and 23% for severe AS (p<.001). Univariable analysis showed that asymptomatic severe AS was significantly associated with greater risk of all-cause mortality (hazard ratio (HR)=3.06, 95% confidence interval (CI)=1.96-4.58, p<.001). After adjustment for age, sex, LVEF, hypertension, dyslipidemia, diabetes, atrial fibrillation, chronic kidney disease, and coronary artery disease, asymptomatic severe AS was an independent predictor of all-cause mortality (HR=3.16, 95% CI=1.97-4.88, p<.001). CONCLUSION: Asymptomatic severe AS has a major effect on prognosis even in very old adults.

Brap2 functions as a cytoplasmic retention protein for p21 during monocyte differentiation.

The cell cycle inhibitor p21 plays an important role in monocytic cell differentiation, during which it translocates from the nucleus to cytoplasm. This process involves the negative regulation of the p21 nuclear localization signal (NLS). Here, we sought to determine the relationship between the cytoplasmic translocation of p21 and another molecule, Brap2, a cytoplasmic protein which binds the NLS of BRCA1 and was recently reported to inactivate KSR in the Ras-activating signal pathway under the name of IMP. We report that p21 and Brap2 directly interact, both in vitro and in vivo, in a manner requiring the NLS of p21 and the C-terminal portion of Brap2. When it is cotransfected with Brap2, p21 is expressed in the cytoplasm. Monocytic differentiation of the promyelomonocytic cell lines U937 and HL60 is associated with the upregulation of Brap2 expression concomitantly with the upregulation and cytoplasmic relocalization of p21. Our results underscore the role played by Brap2 in the process of cytoplasmic translocation of p21 during monocyte differentiation.

Severe Hallux Valgus With Coalition of the Hallux Sesamoids Treated With Modified Lapidus Procedure: A Case Report.

Coalition of the hallux sesamoids is an extremely rare condition. To our knowledge, only 1 case report has been published. We report a case of severe hallux valgus deformities with coalitions of the hallux sesamoids. The coalitions themselves were asymptomatic; however, this severe hallux valgus deformity needed to be surgically treated. The hallux sesamoids in both feet appeared to be fused and heart shaped on anteroposterior radiographs and dumbbell shaped on axial radiographs. It is known that postoperative incomplete reduction of the medial sesamoids can be a risk factor for the recurrence of hallux valgus. The computed tomography scan demonstrated a groove in the bottom of the center of the heart-shaped sesamoid. The flexor hallucis longus tendon was located in the groove. Therefore, a modified Lapidus procedure was performed considering the medial half of the heart-shaped sesamoid as the medial sesamoid. Although delayed union occurred, successful correction of the deformity was achieved. LEVELS OF EVIDENCE: Level IV.

Effectiveness of Percutaneous Proximal Closing Wedge Osteotomy With Akin Osteotomy to Correct Severe Hallux Valgus Determined by Radiographic Parameters.

Some authors reported the results from percutaneous distal metatarsal osteotomy for hallux valgus recently. On the other hand, there are few reports of percutaneous proximal metatarsal osteotomy. The purpose of the present study was to evaluate the radiographic results of percutaneous proximal closing wedge osteotomy with Akin osteotomy for correction of severe hallux valgus and increasing longitudinal arch height. Consecutive 17 feet (mean age = 70.8 years) were investigated. The mean follow-up was 22 months. Excision of medial eminence, distal soft tissue release, and Akin osteotomy were all performed percutaneously and concurrently. Weight-bearing anteroposterior and lateral radiographs of the feet were acquired preoperatively and at final follow-up. On the anteroposterior radiographs, hallux valgus angle, intermetatarsal angle, and first metatarsal shortening were measured. On the lateral radiographs, talometatarsal angle, calcaneal pitch angle, and first metatarsal dorsiflexion were measured. The average improvements in hallux valgus angle and intermetatarsal angle were 27.6° and 9.9°, respectively. The average first metatarsal shortening was 2.7 mm. The first metatarsal dorsiflexion improved by 2.2°; however, other parameters did not improve significantly. In conclusion, percutaneous proximal closing wedge osteotomy with Akin osteotomy corrects severe hallux valgus; however, the procedure does not increase the medial longitudinal arch. LEVELS OF EVIDENCE: Therapeutic, Level IV: Case series.

Marked prevention of tumor growth and metastasis by a novel immunosuppressive agent, FTY720, in mouse breast cancer models.

FTY720 is a unique immunosuppressive agent that exerts its activity by inducing apoptosis in lymphocytes. We conducted the present study to investigate the effects of FTY720 on cancer growth and metastasis, as well as its mechanism of action. In vitro treatment with FTY720 induced dramatic cancer cell apoptosis in a mouse breast cancer cell line, JygMC(A). Electron microscopy revealed distinct changes on the cell surface with decreased filopodias and microvilli in cancer cells treated with FTY720 at 2 microM and clear evidence of apoptosis at 10 microM. Interestingly, the effect of FTY720 was significantly less in the normal fibroblasts than in the cancer cells, indicating greater susceptibility of cancer cells to the agent. We then tested the in vivo effect of FTY720 in a mouse breast cancer model created by inoculating JygMC(A) cells (s.c.) in the flank region of BALB/c-nu/nu mice at three different dosages (2, 5, and 10 mg/kg/day; n = 30/group). Tumor growth was markedly suppressed at a dosage of 5 mg/kg or more without notable side effects. In addition, tumor metastasis, which was dramatically evident in control mice, was significantly prevented even at a low dose (2 mg/kg/day), resulting in a significant prolongation of animal survival. These data led us to additionally investigate the mechanism of action, especially the prevention of metastasis at a low dose. FTY720 treatment at 2 microM caused a remarkable cytoskeletal change with deformed and decreased filopodias in cancer cells. In addition, it significantly decreased the ability of cancer cells to adhere and migrate to extracellular matrix components, and markedly reduced the expression of integrins on the cancer cell surface. These results indicate that FTY720 is a potent anticancer agent that induces cancer cell apoptosis and is markedly effective for prevention of metastasis. The changes of cellular structure with reduction of integrin expression may be one of its underlying mechanisms of action.

An Isolated Chronic Osteochondral Fracture of the Third Metatarsal Head Treated With Bioabsorbable Pins: A Case Report.

An osteochondral fracture of the metatarsal head is generally the result of direct trauma and is associated with additional proximal fractures of the medially adjacent metatarsals. An isolated osteochondral fracture of the metatarsal head is extremely rare, with only 10 published case reports. Open reduction and internal fixation was performed in 6 of 10 cases and in 2 of 4 chronic cases, each with different implants previously. The mechanism of injury was considered to be shear force in 5 of the reported cases. It is difficult to achieve and maintain reduction in these cases, and the best method for treatment of this type of fracture is unclear. We report a case of an isolated chronic osteochondral fracture of the third metatarsal head in a 14-year-old boy. The boy reported pain in his third metatarsophalangeal joint on dismounting from a bicycle 2 months prior to presentation. He could play soccer at first; however, the pain worsened after running long distances. Therefore, he was referred to our department. Magnetic resonance imaging revealed an osteochondral fracture of the metatarsal head without Freiberg's disease. We achieved a good result 12 months postoperatively by using open reduction and internal fixation with 2 bioabsorbable thread pins. LEVELS OF EVIDENCE: Therapeutic, Level IV: Case study.

Redox factor-1/APE suppresses oxidative stress by inhibiting the rac1 GTPase.

Oxidative stress triggered by many environmental and clinical insults results in cellular injury and death. The small GTPase rac1 promotes oxidative stress via the production of reactive oxygen species (ROS). In turn, the homeostatic response to such stress includes up-regulation of the dual function reducing protein/DNA repair enzyme APE/redox factor-1(ref-1). In this report we explore the function and relationship between ref-1 and rac1 in the setting of oxidative stress triggered by re-oxygenation/reperfusion. In a model of mouse hepatic ischemia/reperfusion (I/R), recombinant adenoviral overexpression of ref-1 resulted in suppression of reperfusion-stimulated oxidative stress, NF-kB induction, apoptosis, and acute injury, whereas down-regulation of endogenous ref-1 by adenoviral expression of antisense ref-1 led to an increase in these reperfusion-induced parameters. Ref-1 also mitigated ROS production induced by adenoviral expression of an active form of rac1. Finally, overexpression of ref-1 in primary hepatocytes suppressed reoxygenation-stimulated rac1 activity. This work demonstrates a novel function of ref-1 in inhibition of rac1 activity, and rac1-mediated oxidative stress and injury.

Catalytic RAG1 mutants obstruct V(D)J recombination in vitro and in vivo.

To generate severe combined immunodeficient (SCID) livestocks for xenotransplantation, we have attempted to generate a SCID phenotype without gene knockout. Based on the reported mouse RAG1 mutants, we constructed the corresponding rabbit RAG1 mutants by mutagenesis of three residues within the catalytic domain: D602A, D710A, and E964A. As expected, these mutants each exhibited no catalytic activity on artificial substrates and inhibited recombination by the wild type RAG1. Moreover, replacement of the N-terminus of RAG1 with enhanced green fluorescent protein (EGFP) greatly increased protein stability, and the triple mutant RAG1 showed a twofold increase in its ability to inhibit wild type activity in vitro. We generated mice transgenic for the latter mutant to assess its effect on V(D)J recombination in vivo. Serum IgM levels in four out of seven transgenic mice were reduced to approximately 30-50% of control levels in four out of seven transgenic mice. Our results suggest that immunodeficient animals for regenerative medicine could be generated without gene knockout.

Tibialis Spastic Varus Foot With Calcaneonavicular Coalition: A Case Report and Review of the Literature.

UNLABELLED: Peroneal spastic flat foot is a well-known condition usually occurring with tarsal coalition. Conversely, tibialis spastic varus foot is a rare condition, which can be difficult to diagnose. Moreover, tibialis spastic varus foot with calcaneonavicular coalition is extremely rare, with only a few published case reports. Resection of the calcaneonavicular bar is performed in the majority of patients. We report a case of tibialis spastic varus foot with calcaneonavicular coalition in an 11-year-old boy with intellectual disability. His family noticed his right varus foot deformity 1.5 years earlier. There was no obvious history of trauma. The deformity gradually worsened with running. Because conservative treatment failed, resection of the coalition was performed that facilitated a good outcome. In this article, we report our experience of tibialis spastic varus foot with calcaneonavicular coalition and review the English literature of this condition. LEVEL OF EVIDENCE: Therapeutic, Level IV: Case report.

Prolonged survival in rat liver transplantation with mouse monoclonal antibody against an inducible costimulator (ICOS).

BACKGROUND: An inducible costimulator (ICOS), a recently identified costimulatory receptor with a close structural homology to CD28 and CTLA4, is expressed on activated T cells. Interaction with its ligand on antigen-presenting cells stimulates T-cell proliferation to produce a different spectrum of cytokine. The inhibition of ICOS-mediated signal transduction by an anti-ICOS antibody is considered to be capable of protecting against graft rejection in organ transplantation. METHODS: An anti-rat ICOS antibody was intravenously administered into recipients of dark Agouti-to-Lewis liver transplantations. The recipient lymphocytes from mesenteric lymph nodes were harvested on day 7 after transplantation for fluorescence-activated cell sorting analysis, and tissue specimens from the grafts were removed for histologic evaluation. Antigen-specific T-cell proliferation responses were assessed in vitro with anti-ICOS antibody. RESULTS: Monotherapy with the antibody significantly prolonged the graft survival time by inhibiting T-cell activation and its proliferation response. The graft-infiltrating cells, both CD4 and CD8 T cells, were not completely reduced even when rats were administered the antibody, whereas the expression of ICOS almost completely disappeared in these cells. CONCLUSIONS: T-cell activation through the ICOS costimulatory pathway plays an important role in graft rejection, and manipulating its pathway is an effective method for modulating transplantation immunity.

A novel immunosuppressive drug, FTY720, prevents the cancer progression induced by cyclosporine.

Cyclosporine A (CsA), the most frequently used immunosuppressive drug, has been reported to induce cancer by a cell-autonomous mechanism. Herein, we report that FTY720, a novel immunosuppressant, prevents CsA-induced alterations in both morphology and cell motility at a low concentration (0.1 microM) and induces the CsA-treated cancer cells to undergo apoptosis at a higher concentration (more than 5 microM). The inhibitory activity of FTY720 is unrelated to the decrease of TGF-beta. We believe that a combination treatment with FTY720 and CsA not only results in a synergistic effect on allografts, but also, reduces the incidence of cancer in transplant patients.

Simultaneous blockade of co-stimulatory signals, CD28 and ICOS, induced a stable tolerance in rat heart transplantation.

An inducible co-stimulator (ICOS), a recently identified co-stimulatory receptor with a close structural homology of CD28 and CTLA4, is expressed on activated T cells. Anti-ICOS antibody was demonstrated to be effective on prolongation of graft survival after liver transplantation in rats. In this study, we investigated the potency of tolerance induction using the antibody combined with a recombinant adenovirus vector containing CTLA-4Ig cDNA (AdCTLA-4Ig) in rat heart transplantation model. Using a DA-to-Lewis rat heart transplantation model, an anti-rat ICOS antibody and AdCTLA-4Ig were simultaneously administered i.v. into recipients. The tissue specimens from the grafts were removed on various days after transplantation for histological evaluation. Donor-strain skin and heart grafts, and third-party heart allografts were challenged in the recipients with a long-term surviving graft. Splenocytes from the tolerance-induced recipients were used for adoptive transfer study. Anti-ICOS antibody alone did not prolong the survival of heart allograft. AdCTLA-4Ig monotherapy significantly prolonged the survival of heart allograft (Group 4). With a combination of Anti-ICOS antibody and AdCTLA-4Ig, all recipients were resulted in a long-term allograft acceptance for more than 200 days (Group 8). When challenged donor-strain skin grafts in the tolerant rats of Group 4, the skin was rejected, which also lead to a rejection of primary heart allografts. The recipients in Group 8 also rejected donor-strain skin grafts with no rejection of the primary heart grafts. These recipients accepted secondary heart grafts from donor-strain but not third-party. In Group 8 long-term survival recipients showed a high population of CD4+CD25+ regulatory T cell in peripheral blood, and in adoptive transfer study subtraction of these CD4+CD25+ T cells accelerate the rejection of heart graft in secondary irradiated recipients. The present results demonstrated that anti-ICOS antibody combined with AdCTLA-4Ig potently induces a stable immune tolerance after heart allografting in rat, which is mediated by the induction of CD4+CD25+ regulatory T cells. This strategy may be attractive for clinical employment to induce transplantation tolerance.

Prolongation of liver xenograft survival by adenovirus vector-mediated CTLA-4Ig gene transfer.

Cytotoxic T lymphocyte-associated antigen-4/immunoglobulin fusion products (CTLA-4Ig), a structural homologue of CD28, has been shown to inhibit cellular and humoral immune responses. In this study, we investigated the efficacy of an adenovirus vector containing the CTLA-4Ig gene (AdCTLA-4Ig) on recipient survival after hamster-to-rat liver xenografting. AdCTLA-4Ig was administrated intravenously immediately after grafting. Gene expression was achieved a maximum of 7 days after vector injection and continued for more than 4 weeks. The proportion of CD25(+) T-cells in recipient lymph nodes was significantly reduced 7 days after administration of AdCTLA-4Ig, compared to a group given an adenoviral vector containing LacZ gene (AdLacZ) or to an untreated control group. AdCTLA-4Ig markedly reduced CD2(+) T-cell infiltration into the graft and significantly prolonged recipient survival time (9.2+/-4.12 days), compared to the untreated group (5.4+/-0.78 days) (P<0.001) and the AdLacZ-treated group (5.2+/-0.28 days) (P<0.001). These results indicate that a blockade of T-cell co-stimulation by AdCTLA-4Ig inhibited T-cell activation and attenuated CD2(+) T-cell infiltration into the xenograft, resulting in significant prolongation of recipient survival time. Thus, AdCTLA-4Ig therapy may provide a novel approach to immune regulation.

Transplantation of amniotic epithelial cells into fetal rat liver by in utero manipulation.

It has been hoped that amniotic epithelial cells would be a gene carrier to neural and hepatic tissue, because of 1) the presence of neural and hepatic stem-like cells, 2) the ability to cryopreserve them, 3) long-term survival in the transplanted site, and 4) few ethical problems concerning procurement. But transplantation of a sufficient number of cells to adult tissue needs large-scale cell supply and may lead to vascular embolism. We attempted transplantation of amniotic epithelial cells into fetal liver, because 1) the fetal liver is at the proliferative stage, 2) the number of cells required is small, and 3) the fetal stage is advantageous for the induction of immunological tolerance. Amniotic epithelial cells from day 18.5-20.5 fetuses were transfected with adenoviral AdlacZ and harvested to inject into fetal rat liver of the syngeneic strain (day 18.5-20.5). The efficacy of cell transplantation into the liver increased in the order: intraplacental < intraumbilical vein < intrahepatic route. LacZ-transfected amniotic cells (1-8 x 10(5) cells), hepatocytes (5 x 10(5) cells), or AdlacZ vector solution (1.7 x 10(7) pfu) were injected through the uterine membrane into the liver. Transplanted cells formed a cellular mass and survived for up to 14 days after birth, whereas lacZ-transfected cells were rapidly decreased after the injection of AdlacZ vector or rat hepatocytes as a gene carrier so that the use of amniotic epithelial cells as a gene carrier will result in long-term expression of exogenous genes in the liver.

Selective cancer cell apoptosis induced by FTY720; evidence for a Bcl-dependent pathway and impairment in ERK activity.

BACKGROUND: FTY720 is a unique immunosuppressant that induces apoptosis in activated lymphocytes, but not in other hematopoietic cells. We conducted the present study to investigate its anticancer effect and molecular pathway in inducing apoptosis using murine breast cancer models. MATERIALS AND METHODS: The difference in drug susceptibility to FTY720 between cancer cells and non-cancer cells was examined by MTT assay and cell growth assay. FTY720-induced apoptosis was determined by electron microscopy and DNA electrophoresis, and its molecular pathway was evaluated by Western blot analysis. We then tested in vivo the effect of this agent using two murine breast cancer models. RESULTS: FTY720 treatment induced selective cancer cell apoptosis in vitro at a concentration of less than 10 microM. In vivo tumor growth was significantly prevented with induction of apoptosis in both models without any severe systemic adverse reactions. The evaluation of intracellular protease activity demonstrated that FTY720-induced apoptosis was mediated by a Fas-independent, Bcl-associated signal transduction pathway. Inhibition of extracellular signal-regulated kinase (ERK) activity may be involved in its underlying mechanism of action. CONCLUSION: FTY720 may be a promising candidate for a new anticancer therapy, which potentially induces selective apoptosis in cancer cells.