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CD147 is an immunoglobulin-like receptor that is highly expressed in various cancers and involved in the growth, metastasis, and activation of inflammatory pathways via interactions with various functional molecules, such as integrins, CD44, and monocarboxylate transporters. Through screening of CD147-targeting antibodies with antitumor efficacy, we discovered a novel rat monoclonal antibody #147D. This humanized IgG4-formatted antibody, h4#147D, showed potent antitumor efficacy in xenograft mouse models harboring the human PDAC cell line MIA PaCa-2, HCC cell line Hep G2, and CML cell line KU812, which featured low sensitivity to the corresponding standard-of-care drugs (gemcitabine, sorafenib, and imatinib, respectively). An analysis of tumor cells derived from MIA PaCa-2 xenograft mice treated with h4#147D revealed that cell surface expression of CD147 and its binding partners, including CD44 and integrin α3β1/α6β1, was significantly reduced by h4#147D. Inhibition of focal adhesion kinase (FAK), activation of multiple stress responsible signal proteins such as c-JunN-terminal kinase (JNK) and mitogen-activated protein kinase p38 (p38MAPK), and expression of SMAD4, as well as activation of caspase-3 were obviously observed in the tumor cells, suggesting that h4#147D induced tumor shrinkage by inducing multiple stress responsible signals. These results suggest that the anti-CD147 antibody h4#147D offers promise as a new antibody drug candidate.

Supplementary Materials and Methods

Supplementary Figure S1. Epitope mapping of clones A-D by alanine scanning mutagenesis. Supplementary Figure S2. Analysis of the selectivity of ET1402L1 (clone B) towards AFP158. Supplementary Figure S3. Analysis of the potential risk for cross-presentation of AFP158 by normal cells. Supplementary Figure S4. Co-incubation of AFP-CAR T cells with antigen-positive cells induces production and release of cytokines. Supplementary Figure S5. AFP-CAR T cells can penetrate laminin-rich Matrigel and specifically kill AFP+/A2+ cells in vitro. Supplementary Figure S6. Representative quality control tests performed on each batch of AFP-CAR T cells manufactured for in vivo studies. Supplementary Figure S7. T cell subset analysis to determine distribution of TNaive, TCM TEM, and TEffector cell populations. Supplementary Figure S8. Analysis of human CD3+ T cells in Hep G2 s.c. tumors from i.v.- treated mice. Supplementary Figure S9. Determination of the number of AFP158/HLA-A*02:01 complexes expressed on the surface of liver cancer cells. Supplementary Table S1. Chimeric mouse IgG1 antibody binding affinity to AFP158/HLAA* 02:01.

Figure Legends for Sup. Figures 1-9 and Sup. Table 1

Purpose: The majority of tumor-specific antigens are intracellular and/or secreted and therefore inaccessible by conventional chimeric antigen receptor (CAR) T-cell therapy. Given that all intracellular/secreted proteins are processed into peptides and presented by class I MHC on the surface of tumor cells, we used alpha-fetoprotein (AFP), a specific liver cancer marker, as an example to determine whether peptide–MHC complexes can be targets for CAR T-cell therapy against solid tumors.Experimental Design: We generated a fully human chimeric antigen receptor, ET1402L1-CAR (AFP-CAR), with exquisite selectivity and specificity for the AFP158–166 peptide complexed with human leukocyte antigen (HLA)-A*02:01.Results: We report that T cells expressing AFP-CAR selectively degranulated, released cytokines, and lysed liver cancer cells that were HLA-A*02:01+/AFP+ while sparing cells from multiple tissue types that were negative for either expressed proteins. In vivo, intratumoral injection of AFP-CAR T cells significantly regressed both Hep G2 and AFP158-expressing SK-HEP-1 tumors in SCID-Beige mice (n = 8 for each). Moreover, intravenous administration of AFP-CAR T cells in Hep G2 tumor-bearing NSG mice lead to rapid and profound tumor growth inhibition (n = 6). Finally, in an established intraperitoneal liver cancer xenograft model, AFP-CAR T cells showed robust antitumor activity (n = 6).Conclusions: This study demonstrates that CAR T-cell immunotherapy targeting intracellular/secreted solid tumor antigens can elicit a potent antitumor response. Our approach expands the spectrum of antigens available for redirected T-cell therapy against solid malignancies and offers a promising new avenue for liver cancer immunotherapy. Clin Cancer Res; 23(2); 478–88. ©2016 AACR.

Hereditary deafness is clinically and genetically heterogeneous. We investigated deafness segregating as a recessive trait in two families. Audiological examinations revealed an asymmetric mild to profound hearing loss with childhood or adolescent onset. Exome sequencing of probands identified a homozygous c.475G>A;p.(Glu159Lys) variant of CLDN9 (NM_020982.4) in one family and a homozygous c.370_372dupATC;p.(Ile124dup) CLDN9 variant in an affected individual of a second family. Claudin 9 (CLDN9) is an integral membrane protein and constituent of epithelial bicellular tight junctions that form semi-permeable, paracellular barriers between inner ear perilymphatic and endolymphatic compartments. Computational structural modeling predicts that substitution of a lysine for glutamic acid p.(Glu159Lys) alters one of two cis-interactions between CLDN9 protomers. The p.(Ile124dup) variant is predicted to locally misfold CLDN9 and mCherry tagged p.(Ile124dup) CLDN9 is not targeted to the HeLa cell membrane. In situ hybridization shows that mouse Cldn9 expression increases from embryonic to postnatal development and persists in adult inner ears coinciding with prominent CLDN9 immunoreactivity in tight junctions of epithelia outlining the scala media. Together with the Cldn9 deaf mouse and a homozygous frameshift of CLDN9 previously associated with deafness, the two bi-allelic variants of CLDN9 described here point to CLDN9 as a bona fide human deafness gene.

The Technological Competency as Caring in Nursing Instrument-Revised (TCCNI-R) was developed from the Technological Competency as Caring in Nursing theory. To assess the psychometric properties, a four-factor questionnaire comprised the TCCNI-R. Only 590 of 1,322 online questionnaire responses by nurses in 11 general hospitals in Chugoku-Shikoku, Japan, were analyzed and interpreted. Exploratory factor analysis and confirmatory factor analysis via SEM justified construct validity, and Cronbach's alpha coefficient established reliability. Goodness of fit was rejected; however fit index value was accepted. Root mean square of error approximation showed reasonable approximation error. The TCCNI-R showed content validity and reliability in measuring technological competency as expression of caring in nursing.

For more than three decades it has been known, that striatal neurons become hyperactive after the loss of dopamine input, but the involvement of dopamine (DA) D1‐ or D2‐receptor‐expressing neurons has only been demonstrated indirectly. By recording neuronal activity using fluorescent calcium indicators in D1 or D2 eGFP‐expressing mice, we showed that following dopamine depletion, both types of striatal output neurons are involved in the large increase in neuronal activity generating a characteristic cell assembly of particular neurons that dominate the pattern. When we expressed channelrhodopsin in all the output neurons, light activation in freely moving animals, caused turning like that following dopamine loss. However, if the light stimulation was patterned in pulses the animals circled in the other direction. To explore the neuronal participation during this stimulation we infected normal mice with channelrhodopsin and calcium indicator in striatal output neurons. In slices made from these animals, continuous light stimulation for 15 s induced many cells to be active together and a particular dominant group of neurons, whereas light in patterned pulses activated fewer cells in more variable groups. These results suggest that the simultaneous activity of a large dominant group of striatal output neurons is intimately associated with parkinsonian symptoms.

Epilepsy, deafness, onychodystrophy, osteodystrophy and intellectual disability are associated with a spectrum of mutations of human TBC1D24. The mechanisms underlying TBC1D24-associated disorders and the functions of TBC1D24 are not well understood. Using CRISPR-Cas9 genome editing, we engineered a mouse with a premature translation stop codon equivalent to human S324Tfs(*)3, a recessive mutation of TBC1D24 associated with early infantile epileptic encephalopathy (EIEE). Homozygous S324Tfs(*)3 mice have normal auditory and vestibular functions but show an abrupt onset of spontaneous seizures at postnatal day 15 recapitulating human EIEE. The S324Tfs(*)3 variant is located in an alternatively spliced micro-exon encoding six perfectly conserved amino acids incorporated postnatally into TBC1D24 protein due to a micro-exon utilization switch. During embryonic and early postnatal development, S324Tfs(*)3 homozygotes produce predominantly the shorter wild-type TBC1D24 protein isoform that omits the micro-exon. S324Tfs(*)3 homozygotes show an abrupt onset of seizures at P15 that correlates with a developmental switch to utilization of the micro-exon. A mouse deficient for alternative splice factor SRRM3 impairs incorporation of the Tbc1d24 micro-exon. Wild-type Tbc1d24 mRNA is abundantly expressed in the hippocampus using RNAscope in situ hybridization. Immunogold electron microscopy using a TBC1D24-specific antibody revealed that TBC1D24 is associated with clathrin-coated vesicles and synapses of hippocampal neurons, suggesting a crucial role of TBC1D24 in vesicle trafficking important for neuronal signal transmission. This is the first characterization of a mouse model of human TBC1D24-associated EIEE that can now be used to screen for antiepileptogenic drugs ameliorating TBCID24 seizure disorders.

The aim of this study was to investigate 3-dimensional (3D) airway volume in patients with unilateral cleft lip and palate (UCLP) using computed tomography (CT). The study population comprised 15 UCLP patients (UCLP group) scheduled to receive alveolar bone grafts and 15 with impacted teeth (control group). The clinical requirements for a CT scan were met in both groups. Measurements were recorded from 3D reconstructions of Digital Imaging and Communications in Medicine data obtained from the CT images. Airway volume, cross-sectional area, and linear and angular measurements were recorded. Airway volume and cross-sectional area showed no significant difference between the two groups. The narrowest section of the airway in the UCLP group was tighter than that in the control group, however (p=0.017). The results of this study suggest that this difference in the measurements of the narrowest section of the airway is involved in the particular maxillofacial morphology found in UCLP patients.

The vital role of the splicing factor SRRM4 in vestibular and inner hair cells of the ear is inactivation of the gene repressor REST; however, in outer hair cells, SRRM4 is dispensable for REST inactivation, which SRRM3 accomplishes independently., In mechanosensory hair cells (HCs) of the ear, the transcriptional repressor REST is continuously inactivated by alternative splicing of its pre-mRNA. This mechanism of REST inactivation is crucial for hearing in humans and mice. Rest is one of many pre-mRNAs whose alternative splicing is regulated by the splicing factor SRRM4; Srrm4 loss-of-function mutation in mice (Srrm4bv/bv) causes deafness, balance defects, and degeneration of all HC types other than the outer HCs (OHCs). The specific splicing alterations that drive HC degeneration in Srrm4bv/bv mice are unknown, and the mechanism underlying SRRM4-independent survival of OHCs is undefined. Here, we show that transgenic expression of a dominant-negative REST fragment in Srrm4bv/bv mice is sufficient for long-term rescue of hearing, balancing, HCs, alternative splicing of Rest, and expression of REST target genes including the Srrm4 paralog Srrm3. We also show that in HCs, SRRM3 regulates many of the same exons as SRRM4; OHCs are unique among HCs in that they transiently down-regulate Rest transcription as they mature to express Srrm3 independently of SRRM4; and simultaneous SRRM4–SRRM3 deficiency causes complete HC loss by preventing inactivation of REST in all HCs. Thus, our data reveal that REST inactivation is the primary and essential role of SRRM4 in the ear, and that OHCs differ from other HCs in the SRRM4-independent expression of the functionally SRRM4-like splicing factor SRRM3.

We investigated the effects of bicarbonate/lactate-buffered peritoneal dialysis fluid (B/L-PDF) and lactate-buffered PDF (L-PDF) on cell viability and apoptosis, focusing on monocarboxylate transporters (MCTs). MCT-1 transports lactate into cells. Cell viability and apoptosis of human peritoneal mesothelial cells (HPMCs) were examined by water-soluble tetrazolium salt-1 and TUNEL assays, respectively. The relative number of viable HPMCs was significantly decreased by L-PDF at 48 h (8.8 ± 0.4%) compared with cells cultured in M199, but not by B/L-PDF (66.7 ± 1.1%). Apoptosis was markedly induced by L-PDF at 48 h (69.3 ± 16.2%), but not by B/L-PDF (2.6 ± 0.3%). Knockdown of MCT-1 by small interfering RNA (siRNA) attenuated the L-PDF-induced reduction of viable cells and increased apoptosis compared with control siRNA, but MCT-4 knockdown had no effect. B/L-PDF had lesser effects on cell viability and apoptosis of HPMCs compared with L-PDF. These results suggest that B/L-PDF biocompatibility occurs by avoiding the induction of apoptosis in HPMCs.

All members of the NOX protein family contain a unique b-type cytochrome that mediates the electron transport that characterizes the activity of the multicomponent oxidase complexes. Referred to as cytochrome b558, because of its signature spectral absorbance at 558 nm in reduced-minus-oxidized difference spectroscopy, or cytochrome b(-245), because of its very low midpoint potential of -245 mV at pH 7.0, the protein possesses two stacked inequivalent hemes ligated by pairs of histidine residues in membrane helices h3 and h5. In a flavin-dependent fashion, cytochrome b558 shuttles electrons from cytoplasmic NADPH across membranes to molecular oxygen and thereby generates superoxide anion. By performing reduced-minus-oxidized difference spectroscopy and using the millimolar extinction coefficient, E

All members of the NOX protein family contain a unique b-type cytochrome that mediates the electron transport that characterizes the activity of the multicomponent oxidase complexes. Referred to as cytochrome b558, because of its signature spectral absorbance at 558 nm in reduced-minus-oxidized difference spectroscopy, or cytochrome b(-245), because of its very low midpoint potential of -245 mV at pH 7.0, the protein possesses two stacked inequivalent hemes ligated by pairs of histidine residues in membrane helices h3 and h5. In a flavin-dependent fashion, cytochrome b558 shuttles electrons from cytoplasmic NADPH across membranes to molecular oxygen and thereby generates superoxide anion. By performing reduced-minus-oxidized difference spectroscopy and using the millimolar extinction coefficient, E 559-540 nm = 21.6 cm-1 mM-1, one can calculate the amount of cytochrome b558 in intact cells or partially purified membrane preparations. Measurements in samples where cytochrome b558 is relatively high and the presence of unrelated heme-containing proteins low, as in neutrophils, are straightforward. However, low levels of cytochrome b558 expression combined with an abundance of mitochondria and other sources of heme proteins make spectral detection of cytochrome b558 in non-phagocytic cells extremely challenging.

Supramolecular polymers based on ureido-pyrimidinone (UPy) represent a promising class of biocompatible materials for medical applications. Here, the chemical modification effect of UV irradiation, used to sterilize these materials, is studied. Besides anticipated crosslinking effects, UV irradiation causes telechelic UPy-polymers to become fluorescent. UPy-model compounds confirm a relation between UV-induced changes and the UPy-moiety. UV-induced fluorescence and IR-spectral changes are (partially) reversible by heat and/or solvent treatment. The results indicate the presence of at least two distinct UV-induced molecular species. UPy-model compounds with specific tautomeric forms directly relate fluorescence to UPy-enol tautomers. Photo-enolization is hypothesized to occur via an excited-state intermolecular double proton transfer. Changes in UPy-tautomeric equilibrium and crosslinking are factors that influence the dynamics of UPy-based materials. Identification and understanding of such factors will aid in the successful application of these materials, for example as biomaterial in tissue engineering applications.

Remarkable technological advances have been made in the field of medicine in recent years, one result of which is that a prenatal diagnosis of cleft lip and/or cleft palate (CL/P) is now possible. In this situation, it is extremely important to provide the parents with mental care from the moment they are informed. Here, we describe cases of CL/P treated at our hospital and how such a diagnosis and prenatal counseling are handled. A survey was carried out on 4 cases seen at our department between April 2013 and March 2014. Patients are referred to our department from local or our own obstetrics clinics on a prenatal diagnosis of CL/P based on findings from ultrasonography. If the case is a referral from outside, the patient will first be seen at our own obstetrics department. Our department may then be subsequently requested to provide the parents with prenatal counseling. Effort is made to reassure the parents that postnatal support will be provided, right from the start. Next, the multidisciplinary nature of the treatment process is explained. However, only the essential outline is given at first so as to avoid inducing unnecessary anxiety. A response is also given to any questions the parents may have. Our experience of giving such care leads us to believe that improvements are required in the way that explanations and assistance are provided. The number of cases in which prenatal counseling is required is expected to increase in future.

Summary Skilled motor behavior requires bihemispheric coordination, and participation of striatal outputs originating from two neuronal groups identified by distinctive expression of D1 or D2 dopamine receptors. We trained mice to reach for and grasp a single food pellet and determined how the output pathways differently affected forelimb trajectory and task efficiency. We found that inhibition and excitation of D1-expressing spiny projection neurons (D1SPNs) have a similar effect on kinematics results, as if excitation and inhibition disrupt the whole ensemble dynamics and not exclusively one kind of output. In contrast, D2SPNs participate in control of target accuracy. Further, ex vivo electrophysiological comparison of naive mice and mice exposed to the task showed stronger striatal neuronal connectivity for ipsilateral D1 and contralateral D2 neurons in relation to the paw used. In summary, while the output pathways work together to smoothly execute skill movements, practice of the movement itself changes synaptic patterns.

CD147, a type I membrane protein, is highly expressed in various cancers and is involved in the growth, metastasis, and activation of inflammatory signals in cancer cells via interaction with various molecules, such as integrins, CD44, and monocarboxylate transporters. Additionally, the expression of CD147 in tumors positively correlates with poor prognosis in various cancers. Therefore, CD147 is an attractive target for cancer therapy. However, there have been no successful drugs targeting CD147 so far, possibly because it is a multifunctional protein interacting with multiple molecules, making it challenging to discover a relevant CD147-targeting drug candidate by general in vitro screening methodologies. In this study, we developed novel anti-CD147 antibodies, which showed significant antitumor efficacy in immune-deficient mice bearing human pancreatic cancer cell lines, PANC-1 or MIA PaCa-2. Six anti-CD147 antibodies showing in vivo anti-tumor efficacy were obtained; one mouse IgG3, two rat IgG2b, two rat IgG1, and one rat IgG2a antibodies. Analysis of antibody dependent cellular toxicity, antibody-dependent cellular phagocytosis, and complement-dependent cytotoxicity of chimeric and humanized antibodies with human IgG2 or IgG4P derived from the obtained 6 anti-CD147 antibodies, #147A/B/C/D/E/F, suggested no significant immune-effector function for these antibodies. Despite lacking effector function, these antibodies exhibited significant in vivo antitumor activity, suggesting that the mechanism underlying their anti-tumor efficacy did not involve their effector function. We also found that the anti-tumor efficacy of antibodies #147A and #147B positively correlated with SMAD4 protein expression rather than CD147 protein expression in 10 pancreatic cancer xenograft models (correlation coefficient of #147B: anti-tumor efficacy vs. CD147, 0.245; vs. SMAD4, 0.9328). Moreover, the administration of anti-CD147 antibodies upregulated rhoB expression in an anti-CD147 antibody-sensitive SMAD4-positive MIA PaCa-2 graft mouse model. The stable expression of SMAD4 increased the sensitivity to the anti-CD147 antibody, ch4#147A (chimeric IgG4P derived from #147A) in xenograft mice inoculated with the BxPC-3 pancreatic cancer cell line, which is originally negative for SMAD4 and has low sensitivity to anti-CD147 antibodies. These data suggest that the expression of SMAD4 and activation of SMAD signaling are functionally important for susceptibility to anti-CD147 antibody treatment. The humanized anti-CD147 IgG4P antibody, h4#147D, derived from #147D with cross reactivity to monkey CD147, showed superior anti-tumor efficacy with complete tumor reduction, compared to standard anticancer drugs, including gemcitabine, imatinib, and sorafenib, in xenograft mice bearing corresponding cancer cell lines, MIA PaCa-2, KU812 chronic myeloid leukemia, and HepG2 liver cancer cell lines, respectively. No toxicity of h4#147D was observed in cynomolgus monkeys at a single dose of 100 mg/kg. These data suggest that h4#147D might be a novel anti-CD147 antibody, which could induce SMAD activation and tumor shrinkage in multiple xenograft models. Therefore, it has potential as a promising antitumor therapeutic antibody with superior anti-tumor efficacy to existing therapy. Citation Format: Keisuke Fukuchi, Kayoko Nanai, Shoji Yamamoto, Jun Tsukada, Yusuke Totoki, Koichiro Inaki, Masato Ishigami, Naoyuki Makita, Yoko Nakano, Chigusa Yoshimura, Kozo Yoneda, Masato Amano, Kensuke Nakamura, Yoshiyuki Kanari, Yoko Oda, Haruyuki Nishigohri, Rika Nakano, Atsuko Nishida, Kenji Murakami, Yumi Matsui, Naomi Kasanuki, Shoji Midori, Satoko Funo, Sayako Takahashi, Hironobu Komori, Toshiaki Ohtsuka, Toshinori Agatsuma. Novel anti-CD147 antibodies inducing activation of SMAD signaling and tumor shrinkage in intractable cancer models [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr B095. doi:10.1158/1535-7163.TARG-19-B095

Objective: This study investigated the factors associated with antisocial behavior (AB) in children with developmental disorder and effective treatments. Methods: Participants were 110 schoolchildren with developmental disorder and with or without accompanying AB who visited our hospital between October 2009 and October 2012. Among the children with AB, those who exhibited one or more symptoms of conduct disorder (CD) were assigned to the CD subgroup. We examined the background characteristics, past history, type of antisocial behavior, and symptom improvement after treatment in the children with AB and compared the relevant factors with children with developmental disorder without AB. Results: Of the 110 participants, 72 (65.5%) did not exhibit AB and 38 (34.5%) did, 7 (5.5%) of whom fulfilled the criteria for CD. Compared to the children without AB, the children with AB showed a significantly higher occurrence of attention deficit/hyperactivity disorder (AD/HD), maltreatment, institutionalization due to maltreatment, parental mental/psychological problems, and family instability. After medical treatment combined with social-skills training and parental education, 22 of the 38 children with AB showed improved behavior. In the CD subgroup, 4 children were diagnosed with AD/HD and 3 with pervasive developmental disorder, and none of the 7 improved with treatment. Conclusion: AB was associated with AD/HD, maltreatment, institutionalization, parental mental/psychological problems, and family instability. The most effective therapy was parental education. Children with AB need early intervention given that those who already exhibited symptoms of CD showed little improvement with treatment.

Introduction The objectives of this study were (1) to investigate the expressions of interleukin (IL)-17, RANKL (the receptor activator of NF-kappaB ligand), and osteoprotegerin (OPG) in root resorption areas during experimental tooth movement in rats, and (2) to determine the effect of IL-17 on the expressions of RANKL and OPG mRNA from human dental pulp cells. Methods Twelve male 6-week-old Wistar rats were subjected to an orthodontic force of 50 g to induce a mesially tipping movement of the maxillary first molars for 7 days. The expression levels of tartrate resistant acid phosphatase (TRAP), interleukin (IL)-17, IL-17 receptor (IL-17R), receptor activator of nuclear factor-kappa B ligand (RANKL), and OPG proteins were determined in dental pulp by immunohistochemical analysis. Furthermore, the effects of IL-17 on the expressions of RANKL and OPG mRNA were investigated using human dental pulp cells in vitro. Results In the experimental tooth movements in vivo, resorption lacunae with multinucleated cells were observed in the 50-g group. The immunoreactivities for IL-17, IL-17R, and RANKL were detected in dental pulp tissues subjected to the orthodontic force on day 7. Moreover, IL-17 increased the mRNA expression of RANKL from human dental pulp cells in vitro. Conclusions The results of this study suggest that IL-17 and RANKL may be involved in the process of orthodontically induced inflammatory root resorption in dental pulp cells.

The DNA-binding protein REST forms complexes with histone deacetylases (HDACs) to repress neuronal genes in non-neuronal cells. In differentiating neurons, REST is downregulated predominantly by transcriptional silencing. Here we report that post-transcriptional inactivation of REST by alternative splicing is required for hearing in humans and mice. We show that, in the mechanosensory hair cells of the mouse ear, regulated alternative splicing of a frameshift-causing exon into the Rest mRNA is essential for the derepression of many neuronal genes. Heterozygous deletion of this alternative exon of mouse Rest causes hair cell degeneration and deafness, and the HDAC inhibitor SAHA (Vorinostat) rescues the hearing of these mice. In humans, inhibition of the frameshifting splicing event by a novel REST variant is associated with dominantly inherited deafness. Our data reveal the necessity for alternative splicing-dependent regulation of REST in hair cells, and they identify a potential treatment for a group of hereditary deafness cases.

Minimally invasive intervention strategies after myocardial infarction use state-of-the-art catheter systems that are able to combine mapping of the infarcted area with precise, local injection of drugs. To this end, catheter delivery of drugs that are not immediately pumped out of the heart is still challenging, and requires a carrier matrix that in the solution state can be injected through a long catheter, and instantaneously gelates at the site of injection. To address this unmet need, a pH-switchable supramolecular hydrogel is developed. The supramolecular hydrogel is switched into a liquid at pH > 8.5, with a viscosity low enough to enable passage through a 1-m long catheter while rapidly forming a hydrogel in contact with tissue. The hydrogel has self-healing properties taking care of adjustment to the injection site. Growth factors are delivered from the hydrogel thereby clearly showing a reduction of infarct scar in a pig myocardial infarction model.

Objective To quantify negative aesthetic ratings in primary unrepaired cleft lip infants by further refining our previously reported paired comparison method. Design The Thurstone paired comparison method was used to quantify negative aesthetic ratings of plaster facial models selected and ordered according to a table of paired random numbers. Patients, Participants A total of 30 facial models of unrepaired incomplete unilateral cleft lip infants were used in this study. Raters comprised 20 oral surgeons and anesthesiologists. Results Quantification of aesthetic ratings for the 30 face models of unrepaired incomplete unilateral cleft lip infants was obtained. The ratings ranged from 0 to 5.08. Conclusion Quantification of the aesthetic ratings for the 30 face models will be used as objective variables in a multivariate analysis in the third stage of this ongoing study.

Objective Postoperative evaluation of bone formation in the alveolar cleft by computed tomography imaging has been reported. We quantitatively evaluated bone grafts in the alveolar cleft preoperatively and postoperatively using three-dimensional data and superimposition of images. Subjects A total of 12 patients with complete unilateral cleft lip and palate (six left-sided and six right-sided) were studied. Methods Helical computed tomography scans were taken immediately before surgery and at 6 months after surgery and the DICOM files obtained were processed using Mimics and 3-matic software for three-dimensional data analysis. The preoperative and postoperative computed tomography data were superimposed, and the position and length of the unerupted canines and width of the alveolar cleft measured. Results Strong and significant correlations were observed between bone formation in the alveolar cleft bone graft region and preoperative canine position ( r = –.766, p < .01) and canine length ( r = .681, p < .05). Stepwise multiple regression analysis demonstrated that only the preoperative canine position was independently and significantly correlated with bone formation in the alveolar cleft. Conclusion These results indicate that the optimal timing for surgery is when the canine cusp is close to the alveolar plane.

In natural systems, highly synergistic non-covalent interactions among biomolecular components exert mesoscopic control over hierarchical assemblies. We herein present a multicomponent self-assembly strategy to tune hierarchical supramolecular polymer architectures in water using highly affine and directional ureidopyrimidinone-poly-(ethylene glycol)s (UPy-PEG). Using scattering methods and oscillatory rheology, we observe the structural and mechanical regulation of entangled monofunctional UPy-PEG fibrils by cross-linking bifunctional UPy-PEG fibrils. This supramolecular mixing approach opens the door to a range of subtly distinct materials for chemical and biological applications.

A 2-year-and-4-month-old female infant, 12 kg in weight and 90 cm in height fell off from a table, which was about 1 m height with a toothbrush in her mouth without her parents noticing. Urgent CT scan showed that it penetrated the left side of her oropharyngeal wall to the bifurcation of her right carotid artery. According to the initial assessment, carotid artery seemed intact and there seemed to be no sign of CNS involvement. She underwent general anesthesia for further investigation and operation. We could detect vocal code with ease by inserting Glidescope between her tongue and the toothbrush. After the intubation, we administered fentanyl 25 μg rocuronium 15 mg and sevoflulane 3-5% to her, and then she underwent arteriography. The neurosurgeon found no sign of major arterial injury nor traumatic aneurysm nor CNS involvement. She went to the ICU intubated after the removal of the toothbrush. She was extubated 5 days after operation. One of the benefits of the Glidescope is that we can share the visual image, and we chose it this time. When we expect a difficult airway during management for oropharyngeal trauma, we have to consider the way to manage the airway.

Purpose: The majority of tumor-specific antigens are intracellular and/or secreted and therefore inaccessible by conventional chimeric antigen receptor (CAR) T-cell therapy. Given that all intracellular/secreted proteins are processed into peptides and presented by class I MHC on the surface of tumor cells, we used alpha-fetoprotein (AFP), a specific liver cancer marker, as an example to determine whether peptide–MHC complexes can be targets for CAR T-cell therapy against solid tumors. Experimental Design: We generated a fully human chimeric antigen receptor, ET1402L1-CAR (AFP-CAR), with exquisite selectivity and specificity for the AFP158–166 peptide complexed with human leukocyte antigen (HLA)-A*02:01. Results: We report that T cells expressing AFP-CAR selectively degranulated, released cytokines, and lysed liver cancer cells that were HLA-A*02:01+/AFP+ while sparing cells from multiple tissue types that were negative for either expressed proteins. In vivo, intratumoral injection of AFP-CAR T cells significantly regressed both Hep G2 and AFP158-expressing SK-HEP-1 tumors in SCID-Beige mice (n = 8 for each). Moreover, intravenous administration of AFP-CAR T cells in Hep G2 tumor-bearing NSG mice lead to rapid and profound tumor growth inhibition (n = 6). Finally, in an established intraperitoneal liver cancer xenograft model, AFP-CAR T cells showed robust antitumor activity (n = 6). Conclusions: This study demonstrates that CAR T-cell immunotherapy targeting intracellular/secreted solid tumor antigens can elicit a potent antitumor response. Our approach expands the spectrum of antigens available for redirected T-cell therapy against solid malignancies and offers a promising new avenue for liver cancer immunotherapy. Clin Cancer Res; 23(2); 478–88. ©2016 AACR.

We investigated the effects of bicarbonate/lactate-buffered peritoneal dialysis fluid (B/L-PDF) and lactate-buffered PDF (L-PDF) on cell viability and apoptosis, focusing on monocarboxylate transporters (MCTs). MCT-1 transports lactate into cells. Cell viability and apoptosis of human peritoneal mesothelial cells (HPMCs) were examined by water-soluble tetrazolium salt-1 and TUNEL assays, respectively. The relative number of viable HPMCs was significantly decreased by L-PDF at 48 h (8.8 ± 0.4%) compared with cells cultured in M199, but not by B/L-PDF (66.7 ± 1.1%). Apoptosis was markedly induced by L-PDF at 48 h (69.3 ± 16.2%), but not by B/L-PDF (2.6 ± 0.3%). Knockdown of MCT-1 by small interfering RNA (siRNA) attenuated the L-PDF-induced reduction of viable cells and increased apoptosis compared with control siRNA, but MCT-4 knockdown had no effect. B/L-PDF had lesser effects on cell viability and apoptosis of HPMCs compared with L-PDF. These results suggest that B/L-PDF biocompatibility occurs by avoiding the induction of apoptosis in HPMCs.

Along with the introduction of different psycholinguistic and neurobiological research paradigms, we review some outcomes of second language (L2) research. In particular, we discuss the probe recognition, cross-modal lexical priming, self-paced reading, and plausibility judgment tasks, typically used to examine the temporal course of online sentence processing. Moreover, we present neurobiological methods such as behavioral, electro- and magneto-physiological, and hemodynamic measures. In considering the various types of research methods, we review the research on L2 sentence processing from extremely divergent perspectives. Finally, we discuss methodological issues to introduce the status quo of L2 in psycholinguistic and neurobiological domains.