Chlorophyll a and novel synthetic derivatives alleviate atopic dermatitis by suppressing Th2 cell differentiation via IL-4 receptor modulation.

Atopic dermatitis (AD) treatment has largely relied on non-specific broad immunosuppressants despite their long-term toxicities until the approval of dupilumab, which blocks IL-4 signaling to target Th2 cell responses. Here, we report the discovery of compound 4aa, a novel compound derived from the structure of chlorophyll a, and the efficacy of chlorophyll a to alleviate AD symptoms by oral administration in human AD patients. 4aa downregulated GATA3 and IL-4 in differentiating Th2 cells by potently blocking IL-4 receptor dimerization. In the murine model, oral administration of 4aa reduced the clinical severity of symptoms and scratching behavior by 76% and 72%, respectively. Notably, the elevated serum levels of Th2 cytokines reduced to levels similar to those in the normal group after oral administration of 4aa. Additionally, the toxicological studies showed favorable safety profiles and good tolerance. In conclusion, 4aa may be applied for novel therapeutic developments for patients with AD.

Isobolographic Analysis of Drug Combinations With Intrathecal BRL52537 (κ-Opioid Agonist), Pregabalin (Calcium Channel Modulator), AF 353 (P2X3 Receptor Antagonist), and A804598 (P2X7 Receptor Antagonist) in Neuropathic Rats.

BACKGROUND: Neuropathic pain should be treated with drug combinations exhibiting multiple analgesic mechanisms of action because the mechanism of neuropathic pain involves multiple physiological causes and is mediated by multiple pathways. In this study, we defined the pharmacological interaction of BRL52537 (κ-opioid agonist), pregabalin (calcium channel modulator), AF 353 (P2X3 receptor antagonist), and A804598 (P2X7 receptor antagonist). METHODS: Animal models of neuropathic pain were established by spinal nerve ligation (SNL) in male Sprague-Dawley rats, and responses to the mechanical stimulation using von Frey filaments were measured. Drugs were administered by intrathecal route and were examined for antiallodynic effects, and drug interactions were evaluated using isobolographic analysis. The mRNA expression levels of pain-related receptors in each spinal cord or dorsal root ganglion of naïve, SNL, and drug-treated SNL rats were evaluated using real-time polymerase chain reaction. RESULTS: Intrathecal BRL52537, pregabalin, AF 353, and A804598 produced antiallodynic effects in SNL rats. In the drug combination studies, intrathecal coadministration of BRL52537 with pregabalin or A804598 exhibited synergistic interactions, and other drugs combinations showed additivity. The rank order of potency was observed as follows: BRL52537 + pregabalin > BRL52537 + A804598 > pregabalin + AF 353 > A804598 + pregabalin > BRL52537 + AF 353 > AF 353 + A804598. Real-time polymerase chain reaction indicated that alterations of P2X3 receptor and calcium channel mRNA expression levels were observed, while P2X7 receptor and κ-opioid receptor expression levels were not altered. CONCLUSIONS: These results demonstrated that intrathecal combination of BRL52537, pregabalin, AF 353, and A804598 synergistically or additively attenuated allodynia evoked by SNL, which suggests the possibility to improve the efficacy of single-drug administration.

Transnasal endoscopic ultrasound-guided reduction of maxillary sinus wall fracture.

Surgical morbidity from open reduction and internal fixation (ORIF) of maxillary sinus wall fracture often surpasses the benefits of ORIF. Hence, the authors devised transnasal endoscopic-assisted reduction of maxillary sinus wall fracture (TERM) without internal fixation as a minimally invasive surgery for maxillary sinus wall fracture. The purpose of this study was to investigate the feasibility of TERM in cadavers and patients. Six cadavers were dissected to evaluate the feasibility of TERM. In addition, 20 patients with maxillary sinus wall fractures who underwent TERM in a tertiary hospital from August of 2013 to December of 2015 were enrolled in this study. Demographic factors, type of anesthesia, computed tomography (CT) scans, clinical characteristics of patients, and patient satisfaction with surgery were analyzed. Cadaveric study showed that endoscopic inferior meatus antrostomy is a feasible method of approaching the maxillary sinus wall in cadavers. In addition, counterforce could be applied to the maxillary sinus wall by pushing packed Vaseline-soaked gauze or using a zygomatic process approach via a Gillies incision. Clinical experience revealed that patients experienced good facial contour restoration postoperatively. The extent of fractured bony segments was reduced on postoperative CT without complications. Patient satisfaction with TERM was greater than that with ORIF (p = 0.031). TERM showed its feasibility in both cadaveric study and clinical study. TERM can be a good alternative to ORIF, especially in patients who are reluctant to undergo a facial incision.

Discovery of 3-Phenyl Indazole-Based Novel Chemokine-like Receptor 1 Antagonists for the Treatment of Psoriasis.

Chemokine-like receptor 1 (CMKLR1)─a G protein-coupled receptor─has functional roles in the immune system and related diseases, including psoriasis and metabolic diseases. Psoriasis is a chronic inflammatory disease characterized by skin redness, scaliness, and itching. In this study, we sought to develop novel CMKLR1 antagonists by screening our in-house GPCR-targeting compound library. Moreover, we optimized a phenylindazole-based hit compound with antagonistic activities and evaluated its oral pharmacokinetic properties in a murine model. A structure-based design on the human CMKLR1 homology model identified S-26d as an optimized compound that serves as a potent and orally available antagonist with a pIC50 value of 7.44 in hCMKLR1-transfected CHO cells. Furthermore, in the imiquimod-induced psoriasis-like mouse model, oral administration of S-26d for 1 week significantly alleviated modified psoriasis area and severity index scores (severity of erythema, scaliness, skin thickness) compared with the control group.

Sustainably Powered, Multifunctional Flexible Feedback Implant by the Bifacial Design and Si Photovoltaics.

Advanced design and integration of functioning devices with secured power is of interest for many applications that require complicated, sophisticated, or multifunctional processes in confined environments such as in human bodies. Here, strategies for design and realization are introduced for multifunctional feedback implants with the bifacial design and silicon (Si) photovoltaics in flexible forms. The approaches provide efficient design spaces for flexible Si photovoltaics facing up for sustainable powering and multiple electronic components for feedback functions facing down for sensing, processing, and stimulating in human bodies. The computational and experimental results including in vivo assessments ensure feasibility of the approaches by demonstrating feedback multifunctions, power-harvesting in milliwatts, and mechanical compatibility for operations in live tissues. This work should useful for wide range of applications that require sustainable power and advanced multifunctions.

Multiplexed imaging of therapeutic cells with multispectrally encoded magnetofluorescent nanocomposite emulsions.

Here, we describe the fabrication of multispectrally encoded nanoprobes, perfluorocarbon (PFC)/quantum dots (QDs) nanocomposite emulsions, which could provide both multispectral MR and multicolor optical imaging modalities. Our strategy exploited the combination of the multispectral MR properties of four different PFC materials and the multicolor emission properties of three different colored CdSe/ZnS QDs. The PFC/QDs nanocomposite emulsions were fabricated by exchanging hydrophobic ligands coated onto CdSe/ZnS QDs using 1H,1H,2H,2H-perfluorooctanethiol, which renders the QDs to be dispersible in the PFC liquids. To provide biocompatibility, the PFC liquids containing QDs were emulsified into aqueous solutions with the aid of phospholipids. The distinct (19)F-based MR images of PFC/QDs nanocomposite emulsions were obtained by selective excitation of the nanocomposite emulsions with magnetic resonance frequency of each PFC, while a specific fluorescence image of them could be selected using appropriate optical filters. The uptake of PFC/QDs nanocomposite emulsions was high in phagocytic cells such as macrophages (90.55%) and dendritic cells (85.34%), while it was low in nonphagocytic T cells (33%). We have also shown that the nanocomposite emulsions were successfully applied to differentially visualize immunotherapeutic cells (macrophages, dendritic cells, and T cells) in vivo. The PFC/QDs nanocomposite emulsions are expected to be a promising multimodality nanoprobe for the multiplexed detection and imaging of therapeutic cells both in vitro and in vivo.

Mapping of cutaneous melanoma by femtosecond laser-induced breakdown spectroscopy.

Surgical excision (Mohs micrographic surgery) is the standard procedure to treat a melanoma, in which an in situ histologic examination of sectioned skin is carried out repeatedly until no cancer cells are detected. The possibility to identify melanoma from the surrounding skin by femtosecond laser-induced breakdown spectroscopy (fs-LIBS) is investigated. For experiments, melanoma induced on a hairless mouse by injection of B16/F10 murine melanoma cell was sampled in the form of frozen tissue sections as in Mohs surgery and analyzed by fs-LIBS (λ = 1030 nm, τ = 550 fs). For analysis, the magnesium signal normalized by carbon intensity was utilized to construct an intensity map around the cancer, including both melanoma and surrounding dermis. The intensity map showed a close match to the optically observed morphological and histological features near the cancer region. The results showed that when incorporated into the existing micrographic surgery procedure, fs-LIBS could be a useful tool for histopathologic interpretation of skin cancer possibly with significant reduction of histologic examination time.

Analysis of major elements in pigmented melanocytic chicken skin using laser-induced breakdown spectroscopy.

The concentration difference of major elements in melanocytic skin with respect to pigmentation level is analysed by laser-induced breakdown spectroscopy (LIBS) to investigate the applicability of LIBS as an in situ feedback tool for selective and complete laser removal of melanocytic skin tissue like nevus. The skin of black silkie chicken which had a characteristic darkly pigmented perifollicular skin surrounded by lightly pigmented extrafollicular skin was used as the sample. The results showed higher LIBS signal intensities of Ca2+ and Mg2+ but lower intensities of Na+ , Cl- and K+ in the perifollicular skin than in the extrafollicular skin, which demonstrated the feasibility to use LIBS as a reliable method to distinguish skin tissues with difference in pigmentation level. Plasma emission of biochemical elements generated with a laser irradiation on melanocytic skin lesion.

Generation of electrical power under human skin by subdermal solar cell arrays for implantable bioelectronic devices.

Medical electronic implants can significantly improve people's health and quality of life. These implants are typically powered by batteries, which usually have a finite lifetime and therefore must be replaced periodically using surgical procedures. Recently, subdermal solar cells that can generate electricity by absorbing light transmitted through skin have been proposed as a sustainable electricity source to power medical electronic implants in bodies. However, the results to date have been obtained with animal models. To apply the technology to human beings, electrical performance should be characterized using human skin covering the subdermal solar cells. In this paper, we present electrical performance results (up to 9.05mW/cm2) of the implantable solar cell array under 59 human skin samples isolated from 10 cadavers. The results indicate that the power densities depend on the thickness and tone of the human skin, e.g., higher power was generated under thinner and brighter skin. The generated power density is high enough to operate currently available medical electronic implants such as pacemakers that require tens of microwatt.

Elemental analysis of tissue pellets for the differentiation of epidermal lesion and normal skin by laser-induced breakdown spectroscopy.

By laser induced breakdown spectroscopy (LIBS) analysis of epidermal lesion and dermis tissue pellets of hairless mouse, it is shown that Ca intensity in the epidermal lesion is higher than that in dermis, whereas Na and K intensities have an opposite tendency. It is demonstrated that epidermal lesion and normal dermis can be differentiated with high selectivity either by univariate or multivariate analysis of LIBS spectra with an intensity ratio difference by factor of 8 or classification accuracy over 0.995, respectively.

Differentiation of cutaneous melanoma from surrounding skin using laser-induced breakdown spectroscopy.

Laser-induced breakdown spectroscopy (LIBS) has the potential to be used as a surgical tool for simultaneous tissue ablation and elemental analysis of the ablated tissue. LIBS may be used to distinguish melanoma lesions from the surrounding dermis based on the quantitative difference of elements within melanoma lesions. Here, we measured the elements in homogenized pellets and real tissues from excised skin samples of melanoma-implanted mice. In addition, statistical analysis of LIBS spectra using principal component analysis and linear discriminant analysis was performed. Our results showed that this method had high detection sensitivity, highlighting the potential of this tool in clinical applications.

Subdermal Flexible Solar Cell Arrays for Powering Medical Electronic Implants.

A subdermally implantable flexible photovoltatic (IPV) device is proposed for supplying sustainable electric power to in vivo medical implants. Electric properties of the implanted IPV device are characterized in live animal models. Feasibility of this strategy is demonstrated by operating a flexible pacemaker with the subdermal IPV device which generates DC electric power of ≈647 µW under the skin.

Myeloid-Derived Suppressor Cells Are Controlled by Regulatory T Cells via TGF-ß during Murine Colitis.

Myeloid-derived suppressor cells (MDSCs) are well known regulators of regulatory T cells (Treg cells); however, the direct regulation of MDSCs by Treg cells has not been well characterized. We find that colitis caused by functional deficiency of Treg cells leads to altered expansion and reduced function of MDSCs. During differentiation of MDSCs in vitro from bone marrow cells, Treg cells enhanced MDSC function and controlled their differentiation through a mechanism involving transforming growth factor-ß (TGF-ß). TGF-ß-deficient Treg cells were not able to regulate MDSC function in an experimentally induced model of colitis. Finally, we evaluated the therapeutic effect of TGF-ß-mediated in-vitro-differentiated MDSCs on colitis. Adoptive transfer of MDSCs that differentiated with TGF-ß led to better colitis prevention than the transfer of MDSCs that differentiated without TGF-ß. Our results demonstrate an interaction between Treg cells and MDSCs that contributes to the regulation of MDSC proliferation and the acquisition of immunosuppressive functions.

Development of visible and NIR imaging equipment for small animals with smart pad.

The portable visible and near-infrared (NIR) imaging equipment for a pre-clinical test with small animals was designed and developed in this paper. The developed equipment is composed of a CCD camera, a focusing lens, an objective lens, a NIR band pass filter and a NIR filter driving motor. An NIR ray is mainly used for imaging equipment because it has high light penetration depth in biological tissue. Therefore, NIR fluorescent agents are available for chemical conjugation to targeting molecules in vivo. This equipment can provide a visible image, NIR image and merged image simultaneously. A communication system was specifically established to check obtained images through a smart pad in real time. It is less dependent on space and time than the conventional system.

A Case of Chronic Bullous Disease of Childhood That Was Reactive to the Antigen of 120 kDa (LAD-1).

Chronic bullous disease of childhood (CBDC) is an autoimmune blistering disease that is characterized by Immunoglobulin A (IgA) deposits at the basement membrane zone. IgA autoantibodies (aAbs) from the serum of patients with CBDC react with antigens of 97 kDa (LABD97) and 120 kDa (LAD-1), and both of which are fragments of the extracellular domain of bullous pemphigoid 180 (BP180, type XVII collagen). The CBDC sera reacts with the immunodominant NC16a domain of BP180, which is the major region recognized by IgG aAbs in patients with bullous pemphigoid. A five-year-old boy presented with multiple pruritic tense blisters on the umbilical and inguinal areas for six weeks. The direct immunofluorescence of the perilesional area demonstrated linear deposits of IgA at the basement membrane zone. Using immunoblotting and an enzyme linked immunosorbent assay (ELISA), we identified the IgA aAbs reactive to antigens with a molecular weight of 120 kDa (LAD-1), which is a fragment of the extracellular domain of BP180.