Kasugamycin Is a Novel Chitinase 1 Inhibitor with Strong Antifibrotic Effects on Pulmonary Fibrosis.

Pulmonary fibrosis is a devastating lung disease with few therapeutic options. CHIT1 (chitinase 1), an 18 glycosyl hydrolase family member, contributes to the pathogenesis of pulmonary fibrosis through the regulation of TGF-ß (transforming growth factor-ß) signaling and effector function. Therefore, CHIT1 is a potential therapeutic target for pulmonary fibrosis. This study aimed to identify and characterize a druggable CHIT1 inhibitor with strong antifibrotic activity and minimal toxicity for therapeutic application to pulmonary fibrosis. Extensive screening of small molecule libraries identified the aminoglycoside antibiotic kasugamycin (KSM) as a potent CHIT1 inhibitor. Elevated concentrations of CHIT1 were detected in the lungs of patients with pulmonary fibrosis. In in vivo bleomycin- and TGF-ß-stimulated murine models of pulmonary fibrosis, KSM showed impressive antifibrotic effects in both preventive and therapeutic conditions. In vitro studies also demonstrated that KSM inhibits fibrotic macrophage activation, fibroblast proliferation, and myofibroblast transformation. Null mutation of TGFBRAP1 (TGF-ß-associated protein 1), a recently identified CHIT1 interacting signaling molecule, phenocopied antifibrotic effects of KSM in in vivo lungs and in vitro fibroblasts responses. KSM inhibits the physical association between CHIT1 and TGFBRAP1, suggesting that the antifibrotic effect of KSM is mediated through regulation of TGFBRAP1, at least in part. These studies demonstrate that KSM is a novel CHIT1 inhibitor with a strong antifibrotic effect that can be further developed as an effective and safe therapeutic drug for pulmonary fibrosis.

Self-assembled Micelle Interfering RNA for Effective and Safe Targeting of Dysregulated Genes in Pulmonary Fibrosis.

The siRNA silencing approach has long been used as a method to regulate the expression of specific target genes in vitro and in vivo. However, the effectiveness of delivery and the nonspecific immune-stimulatory function of siRNA are the limiting factors for therapeutic applications of siRNAs. To overcome these limitations, we developed self-assembled micelle inhibitory RNA (SAMiRNA) nanoparticles made of individually biconjugated siRNAs with a hydrophilic polymer and lipid on their ends and characterized their stability, immune-stimulatory function, and in vivo silencing efficacy. SAMiRNAs form very stable nanoparticles with no significant degradation in size distribution and polydispersity index over 1 year. Overnight incubation of SAMiRNAs (3 µm) on murine peripheral blood mononuclear cells did not cause any significant elaboration of innate immune cytokines such as TNF-α, IL-12, or IL-6, whereas unmodified siRNAs or liposomes or liposome complexes significantly stimulated the expression of these cytokines. Last, the in vivo silencing efficacy of SAMiRNAs was evaluated by targeting amphiregulin and connective tissue growth factor in bleomycin or TGF-ß transgenic animal models of pulmonary fibrosis. Intratracheal or intravenous delivery two or three times of amphiregulin or connective tissue growth factor SAMiRNAs significantly reduced the bleomycin- or TGF-ß-stimulated collagen accumulation in the lung and substantially restored the lung function of TGF-ß transgenic mice. This study demonstrates that SAMiRNA nanoparticle is a less toxic, stable siRNA silencing platform for efficient in vivo targeting of genes implicated in the pathogenesis of pulmonary fibrosis.

An Ultrasensitive, Selective, Multiplexed Superbioelectronic Nose That Mimics the Human Sense of Smell.

Human sensory-mimicking systems, such as electronic brains, tongues, skin, and ears, have been promoted for use in improving social welfare. However, no significant achievements have been made in mimicking the human nose due to the complexity of olfactory sensory neurons. Combinational coding of human olfactory receptors (hORs) is essential for odorant discrimination in mixtures, and the development of hOR-combined multiplexed systems has progressed slowly. Here, we report the first demonstration of an artificial multiplexed superbioelectronic nose (MSB-nose) that mimics the human olfactory sensory system, leading to high-performance odorant discriminatory ability in mixtures. Specifically, portable MSB-noses were constructed using highly uniform graphene micropatterns (GMs) that were conjugated with two different hORs, which were employed as transducers in a liquid-ion gated field-effect transistor (FET). Field-induced signals from the MSB-nose were monitored and provided high sensitivity and selectivity toward target odorants (minimum detectable level: 0.1 fM). More importantly, the potential of the MSB-nose as a tool to encode hOR combinations was demonstrated using principal component analysis.

Polypyrrole nanotube embedded reduced graphene oxide transducer for field-effect transistor-type H2O2 biosensor.

We report a rapid-response and high-sensitivity sensor with specificity toward H2O2 based on a liquid-ion-gated field-effect transistor (FET) using graphene-polypyrrole (PPy) nanotube (NT) composites as the conductive channel. The rGO, PPy, NTs, and nanocomposite materials were characterized using Raman spectroscopy, Fourier transform-infrared (FT-IR) spectroscopy, transmission electron microscopy (TEM), and scanning electron microscopy (SEM). On the basis of these results, a well-organized structure is successfully prepared owing to the specific interactions between the PPy NTs and the rGO sheet. Reliable electrical contacts were developed between the rGO/PPy NTs and the microelectrodes, which remained stable when exposed to the liquid-phase analyte. Liquid-ion-gated FETs composed of these graphene nanocomposites exhibited hole-transport behavior with conductivities higher than those of rGO sheets or PPy NTs. This implies an interaction between the PPy NTs and the rGO layers, which is explained in terms of the PPy NTs forming a bridge between the rGO layers. The FET sensor provided a rapid response in real time and high sensitivity toward H2O2 with a limit of detection of 100 pM. The FET-type biosensing geometry was also highly reproducible and stable in air. Furthermore, the liquid-gated FET-type sensor exhibited specificity toward H2O2 in a mixed solution containing compounds found in biological fluids.

High-performance Hg(2+) FET-type sensors based on reduced graphene oxide-polyfuran nanohybrids.

A new type of field-effect transistor (FET) sensor, based on reduced graphene oxide (rGO)-polyfuran (PF) nanohybrids, was strategically developed. The sensing transducer exhibited a rapid response (<1 s) and high sensitivity (10 pM) in a liquid-ion-gated FET-type Hg(2+) sensor. Excellent Hg(2+) discrimination in heavy metal mixtures was also monitored in real time.

Successful treatment of facet joint synovial cyst through percutaneous rupture: a case report.

Facet joint synovial cysts can cause significant back pain and radiculopathy. Treatment options for symptomatic facet joint synovial cysts include surgical excision, facet joint steroid injections, and cyst aspiration. Herein, we report our experience of successfully rupturing a lumbar facet joint synovial cyst through a percutaneous approach with two needles using forceful pressure under C-arm fluoroscopic guidance. The patient experienced immediate symptom improvement that persisted throughout the 24-month follow-up. Our experience highlights that the volume effect technique is a valuable treatment option for symptomatic facet joint synovial cysts under fluoroscopic guidance.

Anesthetic management using remimazolam in a patient with atrial flutter: a case report.

Remimazolam is a new intravenously administered ultra-short-acting benzodiazepine used in anesthesia or sedation. Remimazolam offers several advantages over other short-acting sedatives, including an organ-independent metabolism and rapid and predictable onset and recovery. Furthermore, remimazolam shows less cardiovascular-inhibitory effects than other anesthetics. Atrial flutter is a form of cardiac arrhythmia that is associated with serious health-related outcomes and a substantial economic burden. Acute onset of atrial flutter can cause cardiac dysfunction, hypotension, and myocardial ischemia. Moreover, patients with atrial flutter are likely to have an increased risk of both atrial fibrillation and stroke. In this case report, a patient with a 1-year history of atrial flutter underwent general anesthesia for robot-assisted laparoscopic prostatectomy. Using continuous remimazolam infusion, anesthesia and surgery were successfully completed without sudden changes in the patient's blood pressure, heart rate, or electrocardiogram. This case report describes the first reported use of remimazolam to induce general anesthesia in a patient with atrial flutter. The findings suggest that remimazolam can reduce the hemodynamic risk during anesthesia in patients with arrhythmias such as atrial flutter, and is a suitable option for anesthesia in patients with arrhythmias.

Mycobacterium tuberculosislpdC, Rv0462, induces dendritic cell maturation and Th1 polarization.

Mycobacterium tuberculosis, the etiological factor of pulmonary tuberculosis, causes significant morbidity and mortality worldwide. Activation of host immune responses for containment of mycobacterial infections involves participation of innate immune cells, such as dendritic cells (DCs). In this study, we demonstrated that the gene encoding lipoamide dehydrogenase C (lpdC) from M. tuberculosis, Rv0462, induce maturation and activation of DCs involved in the MAPKs signaling pathway. Moreover, Rv0462-treated DCs activated naïve T cells, polarized CD4(+) and CD8(+) T cells to secrete IFN-γ in syngeneic mixed lymphocyte reactions, which would be expected to contribute to Th1 polarization of the immune response. Our results suggest that Rv0462 can contribute to the innate and adaptive immune responses during tuberculosis infection, and thus modulate the clinical course of tuberculosis.

Clonorchis sinensis-derived total protein attenuates airway inflammation in murine asthma model by inducing regulatory T cells and modulating dendritic cell functions.

Asthma is characterized by Th2-mediated inflammation, resulting in airway hyperresponsiveness (AHR) through airway remodeling. Recent epidemiological and experimental reports have suggested an inverse relationship between the development of allergy and helminth infections. Infection by Clonorchis sinensis, a liver fluke that resides in the bile duct of humans, is endemic predominantly in Asia including Korea and China. Using a murine model for asthma, we investigated the effects of C. sinensis-derived total protein (Cs-TP) on allergen-induced airway inflammation and the mechanism underlying the protective effects of Cs-TP administration on asthma. Treatment with Cs-TP attenuated OVA-induced airway inflammation and methacholine-induced AHR, as well as eosinophilia development, lymphocyte infiltration into the lung, and goblet cell metaplasia. This protective effect of Cs-TP is associated with markedly reduced OVA-specific IgE and Th1/Th2 cytokine production. Moreover, Cs-TP increased the number of CD4(+)CD25(+)Foxp3(+) regulatory T (Treg) cells as well as their suppressive activity. In fact, proliferation of OVA-restimulated splenocytes was suppressed significantly. Cs-TP also inhibited the expression of such co-stimulatory molecules as CD80, CD86, and CD40 in LPS- or OVA-stimulated dendritic cells (DCs), suggesting that Cs-TP could interfere with the capacity of airway DCs to prime naïve T cells. These data demonstrate the capacity of C. sinensis to ameliorate allergic asthma and broaden our understanding of the paradoxical relationship between the allergic immune response and helminth infection.

Effect of clinical anesthesia preoperative evaluation on the length of preoperative period and total hospitalization of patients undergoing laparoscopic cholecystectomy.

OBJECTIVE: Anesthesia preoperative evaluation clinics (APECs) are currently operating in several South Korean hospitals. While several studies have investigated the impact of APEC operations on the length of total hospital stay (LTHS), few have investigated their impact on the length of preoperative hospital stay (LPHS) for patients. In this study, we aimed to determine whether APEC affected the LPHS and LTHS. METHODS: Data of all patients who underwent surgery at Chungbuk National University Hospital between September 2009 and August 2019 were analyzed retrospectively. All patients who had undergone laparoscopic cholecystectomy over the last 10 years were categorized into two groups: those who visited the APEC (Group A), and those who did not (Group B). The age, sex, American Society of Anesthesiologists physical status score, LPHS, and LTHS of the two groups were compared. RESULTS: The LPHS was 1.03±0.2 days in Group A and 1.61±1.6 days in Group B. The LTHS was 4.77±1.9 days in Group A and 5.63±2.6 days in Group B. The LPHS and LTHS of the two groups differed by 0.58 and 0.9 days, respectively. CONCLUSION: We evaluated the effect of APEC operations on the LPHS and LTHS of inpatients undergoing laparoscopic cholecystectomy and observed a decrease in both the LPHS and LTHS. Understanding and accepting the importance of APEC is significant for physicians and administrators working to improve hospital efficiency and patient outcomes. Further research is needed to investigate the need and benefits of APECs.

Three-dimensional printed polylactic acid scaffold integrated with BMP-2 laden hydrogel for precise bone regeneration.

BACKGROUND: Critical bone defects remain challenges for clinicians, which cannot heal spontaneously and require medical intervention. Following the development of three-dimensional (3D) printing technology is widely used in bone tissue engineering for its outstanding customizability. The 3D printed scaffolds were usually accompanied with growth factors, such as bone morphometric protein 2 (BMP-2), whose effects have been widely investigated on bone regeneration. We previously fabricated and investigated the effect of a polylactic acid (PLA) cage/Biogel scaffold as a carrier of BMP-2. In this study, we furtherly investigated the effect of another shape of PLA cage/Biogel scaffold as a carrier of BMP-2 in a rat calvaria defect model and an ectopic ossification (EO) model. METHOD: The PLA scaffold was printed with a basic commercial 3D printer, and the PLA scaffold was combined with gelatin and alginate-based Biogel and BMP-2 to induce bone regeneration. The experimental groups were divided into PLA scaffold, PLA scaffold with Biogel, PLA scaffold filled with BMP-2, and PLA scaffold with Biogel and BMP-2 and were tested both in vitro and in vivo. One-way ANOVA with Bonferroni post-hoc analysis was used to determine whether statistically significant difference exists between groups. RESULT: The in vitro results showed the cage/Biogel scaffold released BMP-2 with an initial burst release and followed by a sustained slow-release pattern. The released BMP-2 maintained its osteoinductivity for at least 14 days. The in vivo results showed the cage/Biogel/BMP-2 group had the highest bone regeneration in the rat calvarial defect model and EO model. Especially, the bone regenerated more regularly in the EO model at the implanted sites, which indicated the cage/Biogel had an outstanding ability to control the shape of regenerated bone. CONCLUSION: In conclusion, the 3D printed PLA cage/Biogel scaffold system was proved to be a proper carrier for BMP-2 that induced significant bone regeneration and induced bone formation following the designed shape.

Role of Chitinase 3-Like 1 Protein in the Pathogenesis of Hepatic Insulin Resistance in Nonalcoholic Fatty Liver Disease.

A recently discovered human glycoprotein, chitinase 3-like 1 (Chi3L1), may play a role in inflammation, tissue remodeling, and visceral fat accumulation. We hypothesize that Chi3L1 gene expression is important in the development of hepatic insulin resistance characterized by the generation of pAKT, pGSK, and pERK in wild type and Chi3L1 knockout (KO) murine liver following insulin stimulation. The Chi3L1 gene and protein expression was evaluated by Real Time PCR and ELISA; lipid accumulation in hepatocytes was also assessed. To alter Chi3L1 function, three different anti-Chi3L1 monoclonal antibodies (mAbs) were administered in vivo and effects on the insulin signaling cascade and hepatic lipid deposition were determined. Transmission of the hepatic insulin signal was substantially improved following KO of the CHi3L1 gene and there was reduced lipid deposition produced by a HFD. The HFD-fed mice exhibited increased Chi3L1 expression in the liver and there was impaired insulin signal transduction. All three anti-Chi3L1 mAbs partially restored hepatic insulin sensitivity which was associated with reduced lipid accumulation in hepatocytes as well. A KO of the Chi3L1 gene reduced lipid accumulation and improved insulin signaling. Therefore, Chi3L1 gene upregulation may be an important factor in the generation of NAFLD/NASH phenotype.

A rare case of acute meningitis shortly after lumbar selective nerve root block: a case report.

A 57-year-old man underwent lumbar selective nerve root block (SNRB) for low back pain and lower radiating pain caused by left-sided L4 disc herniation. He presented to the emergency department with fever, headache and aggravated low back pain approximately 3 hours after the procedure. Infection was suspected; hence, blood tests, cerebrospinal fluid (CSF) tests, lumbar magnetic resonance imaging, and brain computed tomography were performed. Imaging findings were not suggestive of infection. The CSF was turbid and yellowish with pleiocytosis; however, the CSF culture was negative. Based on these findings, the patient was diagnosed with acute meningitis. Broad-spectrum antibiotics and steroid therapy were initiated considering the patient's age and general condition. From hospital day (HD) 2, fever and headache were reduced and disappeared completely by HD 5. At the last follow-up, 1 month after discharge, the patient had no symptoms. Acute meningitis is associated with a high mortality and neurologic deficits. Hence, timely tests, diagnosis, and treatment are critical for positive outcomes. Symptoms of meningitis following a nerve block generally occur within 24-48 hours after the procedure. This case is notable, as it involved a quicker and more sudden onset of symptoms; meningitis occurred only a few hours after lumbar selective nerve root block.

Oncostatin M induces dendritic cell maturation and Th1 polarization.

Oncostatin M (OSM) is a pleiotropic cytokine and a member of the gp130/IL-6 cytokine family that has been found to be involved in both pro- and anti-inflammatory responses in cell-mediated immunity. Maturation of dendritic cells (DCs) is crucial for initiation of primary immune responses and is regulated by several stimuli. In this study, the role of OSM in the phenotypic and functional maturation of DCs was evaluated in vitro. Stimulation with OSM upregulated the expression of CD80, CD86, MHC class I and MHC class II and reduced the endocytic capacity of immature DCs. Moreover, OSM induced the allogeneic immunostimulatory capacity of DCs by stimulating the production of the Th1-promoting cytokine IL-12. OSM also increased the production of IFN-gamma by T cells in mixed-lymphocyte reactions, which would be expected to contribute to the Th1 polarization of the immune response. The expression of surface markers and cytokine production in DCs was mediated by both the MAPK and NF-kappaB pathways. Taken together, these results indicate that OSM may play a role in innate immunity and in acquired immunity by enhancing DCs maturation and promoting Th1 immune responses.

Outer membrane protein a of Salmonella enterica serovar Typhimurium activates dendritic cells and enhances Th1 polarization.

BACKGROUND: Typhoid, which is caused by Salmonella enterica serovar Typhimurium, remains a major health concern worldwide. Multidrug-resistant strains of Salmonella have emerged which exhibit increased survivability and virulence, thus leading to increased morbidity. However, little is known about the protective immune response against this microorganism. The outer membrane protein (Omp)A of bacteria plays an important role in pathogenesis. RESULTS: We purified OmpA from S. enterica serovar Typhimurium (OmpA-sal) and characterized the role of OmpA-sal in promoting adaptive and innate immune responses. OmpA-sal functionally activated bone marrow-derived dendritic cells by augmenting expression of CD80, CD86, and major histocompatibility complex classes I and II. Interestingly, OmpA-sal induced production of interferon-γ from T cells in mixed lymphocyte reactions, thus indicating Th1-polarizing capacity. The expression of surface markers and cytokine production in dendritic cells was mediated by the TLR4 signaling pathway in a TLR4 Knock-out system. CONCLUSIONS: Our findings suggest that OmpA-sal modulates the adaptive immune responses to S. enterica serovar Typhimurium by activating dendritic cells and driving Th1 polarization, which are important properties to consider in the development of effective S. enterica serovar Typhimurium vaccines and immunotherapy adjuvant.

Chitinase 1 regulates pulmonary fibrosis by modulating TGF-ß/SMAD7 pathway via TGFBRAP1 and FOXO3.

TGF-ß1 is a critical mediator of tissue fibrosis in health and disease whose effects are augmented by chitinase 1 (CHIT1). However, the mechanisms that CHIT1 uses to regulate TGF-ß1-mediated fibrotic responses have not been defined. Here, we demonstrate that CHIT1 enhances TGF-ß1-stimulated fibrotic cellular and tissue responses and TGF-ß1 signaling. Importantly, we also demonstrate that these effects are mediated by the ability of CHIT1 to inhibit TGF-ß1 induction of its feedback inhibitor, SMAD7. CHIT1 also interacted with TGF-ß receptor associated protein 1 (TGFBRAP1) and forkhead box O3 (FOXO3) with TGFBRAP1 playing a critical role in CHIT1 enhancement of TGF-ß1 signaling and effector responses and FOXO3 playing a critical role in TGF-ß1 induction of SMAD7. These pathways were disease relevant because the levels of CHIT1 were increased and inversely correlated with SMAD7 in tissues from patients with idiopathic pulmonary fibrosis or scleroderma-associated interstitial lung disease. These studies demonstrate that CHIT1 regulates TGF-ß1/SMAD7 axis via TGFBRAP1 and FOXO3 and highlight the importance of these pathways in the pathogenesis of pulmonary fibrosis.

Laminin α1 is a genetic modifier of TGF-ß1-stimulated pulmonary fibrosis.

The pathogenetic mechanisms underlying the pathologic fibrosis in diseases such as idiopathic pulmonary fibrosis (IPF) are poorly understood. To identify genetic factors affecting susceptibility to IPF, we analyzed a murine genetic model of IPF in which a profibrotic cytokine (TGF-ß1) was expressed in the lungs of 10 different inbred mouse strains. Surprisingly, the extent of TGF-ß1-induced lung fibrosis was highly strain dependent. Haplotype-based computational genetic analysis and gene expression profiling of lung tissue obtained from fibrosis-susceptible and -resistant strains identified laminin α1 (Lama1) as a genetic modifier for susceptibility to IPF. Subsequent studies demonstrated that Lama1 plays an important role in multiple processes that affect the pulmonary response to lung injury and susceptibility to fibrosis, which include: macrophage activation, fibroblast proliferation, myofibroblast transformation, and the production of extracellular matrix. Also, Lama1 mRNA expression was significantly increased in lung tissue obtained from IPF patients. These studies identify Lama1 as the genetic modifier of TGF-ß1 effector responses that significantly affects the development of pulmonary fibrosis.

Size-controllable ultrathin carboxylated polypyrrole nanotube transducer for extremely sensitive 17ß-estradiol FET-type biosensors.

17ß-Estradiol is known as a steroid hormone in the human body but it is also known as a disruptor that can cause disequilibrium and dysfunction of the human immune system. Recently, there has been much interest in developing biosensors to detect low concentrations of 17ß-estradiol. In this work, size-controllable aptamer conjugated ultrathin carboxylated polypyrrole nanotubes (A-UCPPyNTs) were fabricated as transducers in 17ß-estradiol field-effect transistor (FET)-type biosensors. They were manufactured via a self-degradation method under several different conditions to control the diameter of the nanotubes. For targeting 17ß-estradiol, the binding aptamers were immobilized through covalent bonding on its surface. The resulting A-UCPNT FET-type biosensor demonstrated p-type behavior with outstanding electrical conductivity, and exhibited Ohmic contacts between the samples and electrodes. The smaller diameter (40 nm) of ultrathin carboxylated polypyrrole nanotubes (UCPPyNTs) contributed to the biosensor's enhanced performance by generating a larger surface area, thereby increasing the number of conjugated binding aptamers. In conclusion, the A-UCPPyNT FET-type biosensor showed extremely high sensitivity (∼1 fM) toward 17ß-estradiol, approximately 103 times more sensitive than the results found in other reports. Moreover, the A-UCPPyNT FET-type biosensor showed unique selectivity to the 17ß-estradiol molecule, in addition to outstanding reusability and long-term storage stability (4 weeks of duration achieved in this work). These performances concerned with reusability and stability were achieved by the formation of covalent bonding in the anchorage to the substrate electrode. Thus this study can be effectively applied in biological and environmental fields.