Synthesis and evaluation of antibody-drug conjugates with high drug-to-antibody ratio using dimaleimide-DM1 as a linker- payload.

The notable characteristics of recently emerged Antibody-Drug Conjugates (ADCs) encompass the targeting of Human Epidermal growth factor Receptor 2 (HER2) through monoclonal antibodies (mAbs) and a high ratio of drug to antibody (DAR). The achievements of Kadcyla® (T-DM1) and Enhertu® (T-Dxd) have demonstrated that HER2-targeting antibodies, such as trastuzumab, have shown to be competitive in terms of efficacy and price for development. Furthermore, with the arrival of T-Dxd and Trodelvy®, high-DAR (7-8) ADCs, which differ from the moderate DAR (3-4) ADCs that were formerly regarded as conventional, are being acknowledged for their worth. Following this trend of drug development, we endeavored to develop a high-DAR ADC using a straightforward approach involving the utilization of DM1, a highly potent substance, in combination with the widely recognized trastuzumab. To achieve a high DAR, DM1 was conjugated to reduced cysteine through the simple design and synthesis of various dimaleimide linkers with differing lengths. Using LC and MS analysis, we have demonstrated that our synthesis methodology is uncomplicated and efficacious, yielding trastuzumab-based ADCs that exhibit a remarkable degree of uniformity. These ADCs have been experimentally substantiated to exert an inhibitory effect on cancer cells in vitro, thus affirming their value as noteworthy additions to the realm of ADCs.

Risk Factors for Respiratory Depression Associated with Tramadol Based on the Global Pharmacovigilance Database (VigiBase).

Tramadol, a weak µ-opioid receptor agonist, has been used worldwide for pain management. It is considered to have a favorable safety profile without serious adverse events; however, safety issues of respiratory depression were proposed by regulatory governments. We aimed to examine the risk and contributing factors associated with tramadol-related respiratory depression using a real-world database, VigiBase. Disproportionality analysis of tramadol and tramadol/paracetamol was performed using proportional reporting ratios, reporting odds ratios, and information components for all drugs and opioids. Factors related to respiratory depression, including sex, age, presence of abuse, death, and various concomitant medications, were evaluated. Among 140,721 tramadol reports, respiratory depression was reported in 1126 cases, 81.3% of which were deemed serious. Five adverse events were detected as signals of tramadol-related acute central respiratory depression (ACRD) in 882 reports. A higher proportion of ACRD cases in children and adolescents was observed than all adverse events cases of tramadol. Concomitant users of CYP2D6 inhibitors, opioids, benzodiazepines, and anti-depressant drugs showed a higher proportion in ACRD cases than non-ACRD cases. ACRD was related to drug abuse and death. This pharmacovigilance study, using VigiBase, confirmed a high risk of respiratory depression (a serious, potentially fatal adverse event) secondary to the use of tramadol, especially in pediatric patients, drug abusers, or during concomitant use of opioids, benzodiazepines, or antidepressants.

Effect of Repeated Greater Occipital Nerve Block in Patients with Ocular Neuropathic Pain: A Retrospective Observational Study.

Ocular neuropathic pain (ONP) has various etiologies, and patients have various symptoms. The clinical management of patients with ONP has been debated. We aimed to evaluate the effect of repeated greater occipital nerve block (GONB) on ONP based on convergence in the trigeminocervical complex. In this single-center retrospective study, the medical records of 204 patients who were referred to the pain clinic by the ophthalmology department of our hospital and subsequently underwent repeated GONB for chronic ONP between January 2008 and February 2022 were analyzed. They received GONB every two weeks, up to 10 times. Symptoms of ONP were divided into five categories: eye pain, dysesthesias/allodynia, non-eye pain, visual disturbance, and tearing. The primary outcome of this study was the pain relief scale (PRS) score after repeated injections. The differences and magnitude of decrease in the overall pain relief scale score were statistically significant (estimate = -0.55, p < 0.001). There were two patients who had recurrence of ONP and seven patients who had adverse events. According to our study, repeated GONB can reduce symptom severity in patients with ONP. Therefore, it appears that GONB can be considered a multimodal management method for ONP.

Lysophosphatidic Acid Induces Podocyte Pyroptosis in Diabetic Nephropathy by an Increase of Egr1 Expression via Downregulation of EzH2.

Podocyte damage and renal inflammation are the main features and pathogenesis of diabetic nephropathy (DN). Inhibition of lysophosphatidic acid (LPA) receptor 1 (LPAR1) suppresses glomerular inflammation and improves DN. Herein, we investigated LPA-induced podocyte damage and its underlying mechanisms in DN. We investigated the effects of AM095, a specific LPAR1 inhibitor, on podocytes from streptozotocin (STZ)-induced diabetic mice. E11 cells were treated with LPA in the presence or absence of AM095, and the expression of NLRP3 inflammasome factors and pyroptosis were measured. A chromatin immunoprecipitation assay and Western blotting were performed to elucidate underlying molecular mechanisms. Gene knockdown by transfecting small interfering RNA was used to determine the role of the transcription factor Egr1 (early growth response protein 1) and histone methyltransferase EzH2 (Enhancer of Zeste Homolog 2) in LPA-induced podocyte injury. AM095 administration inhibited podocyte loss, NLRP3 inflammasome factor expression, and cell death in STZ-induced diabetic mice. In E11 cells, LPA increased NLRP3 inflammasome activation and pyroptosis via LPAR1. Egr1 mediated NLRP3 inflammasome activation and pyroptosis in LPA-treated E11 cells. LPA decreased H3K27me3 enrichment at the Egr1 promoter in E11 cells by downregulating EzH2 expression. EzH2 knockdown further increased LPA-induced Egr1 expression. In podocytes from STZ-induced diabetic mice, AM095 suppressed Egr1 expression increase and EzH2/H3K27me3 expression reduction. Collectively, these results demonstrate that LPA induces NLRP3 inflammasome activation by downregulating EzH2/H3K27me3 and upregulating Egr1 expression, resulting in podocyte damage and pyroptosis, which may be a potential mechanism of DN progression.

Investigation of highly luminescent inorganic perovskite nanocrystals synthesized using optimized ultrasonication method.

All-inorganic halide perovskite nanocrystals are next-generation materials with excellent optical and semiconductor properties suitable for display applications. In this study, we introduce an optimized ultrasonication method for the high-capacity synthesis of highly luminescent inorganic perovskite nanocrystals. After the synthesis of CsPbBr3 with superior optical performance by ultrasonication method, halide anion exchange was performed to tune the stable emission wavelength over the entire visible range. In particular, the maximum photoluminescence wavelengths of the red and green perovskite nanocrystals were appropriate for light-emitting diode applications, and their full-width-at-half-maximum were very narrow, showing outstanding color purity. The materials also had excellent thermal and photo-stability, which is a necessary requirement for perovskite nanocrystal/organic light-emitting diode hybrid device applications. We formulated uniformly stable perovskite nanocrystal inks and optimized their physical and rheological properties for successful inkjet-printing. Finally, we fabricated a hybrid device with a color conversion layer based on the red and green perovskite nanocrystals synthesized using the optimized ultrasonication and halide-ion-exchange methods. The color reproduction range of the fabricated devices was 27.3 % wider than that of the National Television System Committee values, indicating very vivid colors.

Lysophosphatidic Acid Promotes Epithelial-Mesenchymal Transition in Kidney Epithelial Cells via the LPAR1/MAPK-AKT/KLF5 Signaling Pathway in Diabetic Nephropathy.

The epithelial-mesenchymal transition (EMT) is a differentiation process associated with fibrogenesis in diabetic nephropathy (DN). Lysophosphatidic acid (LPA) is a small, naturally occurring glycerophospholipid implicated in the pathogenesis of DN. In this study, we investigated the role of LPA/LPAR1 signaling in the EMT of tubular cells as well as the underlying mechanisms. We observed a decrease in E-cadherin and an increase in vimentin expression levels in the kidney tubules of diabetic db/db mice, and treatment with ki16425 (LPAR1/3 inhibitor) inhibited the expression of these EMT markers. Ki16425 treatment also decreased the expression levels of the fibrotic factors fibronectin and alpha-smooth muscle actin (α-SMA) in db/db mice. Similarly, we found that LPA decreased E-cadherin expression and increased vimentin expression in HK-2 cells, which was reversed by treatment with ki16425 or AM095 (LPAR1 inhibitor). In addition, the expression levels of fibronectin and α-SMA were increased by LPA, and this effect was reversed by treatment with ki16425 and AM095 or by LPAR1 knockdown. Moreover, LPA induced the expression of the transcription factor, Krüppel-like factor 5 (KLF5), which was decreased by AM095 treatment or LPAR1 knockdown. The expression levels of EMT markers and fibrotic factors induced by LPA were decreased upon KLF5 knockdown in HK-2 cells. Inhibition of the extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and serine-threonine kinase (AKT) pathways decreased LPA-induced expression of KLF5 and EMT markers. In conclusion, these data suggest that LPA contributes to the pathogenesis of diabetic nephropathy by inducing EMT and renal tubular fibrosis via regulation of KLF5 through the LPAR1.

Synthesis and Characterization of Diketopyrrolopyrrole-Based Aggregation-Induced Emission Nanoparticles for Bioimaging.

Conventional fluorescent dyes have the property of decreasing fluorescence due to aggregation-caused quenching effects at high concentrations, whereas aggregation-induced emission dyes have the property of increasing fluorescence as they aggregate with each other. In this study, diketopyrrolopyrrole-based long-wavelength aggregation-induced emission dyes were used to prepare biocompatible nanoparticles suitable for bioimaging. Aggregation-induced emission nanoparticles with the best morphology and photoluminescence intensity were obtained through a fast, simple preparation method using an ultrasonicator. The optimally prepared nanoparticles from 3,6-bis(4-((E)-4-(bis(40-(1,2,2-triphenylvinyl)-[1,10-biphenyl]-4-yl)amino)styryl)phenyl)-2,5-dihexyl-2,5-dihydropyrrolo[3,4-c]pyrrole-1,4-dione (DP-R2) with two functional groups having aggregation-induced emission properties and additional donating groups at the end of the triphenylamine groups were considered to have the greatest potential as a fluorescent probe for bioimaging. Furthermore, it was found that the tendency for aggregation-induced emission, which was apparent for the dye itself, became much more marked after the dyes were incorporated within nanoparticles. While the photoluminescence intensities of the dyes were observed to decrease rapidly over time, the prepared nanoparticles encapsulated within the biocompatible polymers maintained their initial optical properties very well. Lastly, when the cell viability test was conducted, excellent biocompatibility was demonstrated for each of the prepared nanoparticles.

Investigation of the Chemical Structure of Ultra-Thin Polyimide Substrate for the Xenon Flash Lamp Lift-off Technology.

Lift-off is one of the last steps in the production of next-generation flexible electronics. It is important that this step is completed quickly to prevent damage to ultrathin manufactured electronics. This study investigated the chemical structure of polyimide most suitable for the Xe Flash lamp-Lift-Off process, a next-generation lift-off technology that will replace the current dominant laser lift-off process. Based on the characteristics of the peeled-off polyimide films, the Xe Flash lamp based lift-off mechanism was identified as photothermal decomposition. This occurs by thermal conduction via light-to-heat conversion. The synthesized polyimide films treated with the Xe Flash lamp-Lift-Off process exhibited various thermal, optical, dielectric, and surface characteristics depending on their chemical structures. The polyimide molecules with high concentrations of -CF3 functional groups and kinked chemical structures demonstrated the most promising peeling properties, optical transparencies, and dielectric constants. In particular, an ultra-thin polyimide substrate (6 µm) was successfully fabricated and showed potential for use in next-generation flexible electronics.

Multiple Ossicular Dislocation Including Stapediovestibular Dislocation Presenting with Conductive Hearing Loss.

Temporal bone trauma can cause hearing loss and in case of prolonged conductive hearing loss, traumatic ossicular injury should be considered. Separation of the incudostapedial joint is the most common lesion, and stapediovestibular dislocation is relatively rare but can easily cause perilymphatic fistula. Here, we report a very rare case of external stapediovestibular dislocation after trauma, ending up with successful surgical outcome. A 27-year-old man with non-progressive hearing loss on the right side since childhood visited the clinic. Audiogram showed a conductive hearing loss with air-bone gap of 55 dB on the right side. Temporal bone CT revealed the disruption of ossicular chain. An exploratory tympanotomy identified multiple ossicular disruptions including external stapediovestibular dislocation with shiny fibrous membrane sealing the oval window. Ossicular chain reconstruction was performed using the total ossicular replacement prosthesis of titanium. A postoperative audiogram showed a recovery of air-bone gap less than 10 dB. To the best of our knowledge, this is the first case of external long-standing stapediovestibular dislocation, with oval window completely sealed with fibrous membrane, ending up with successful hearing recovery by surgery. This case would help dealing with such condition which can be encountered in the clinic.

Preimplant Hearing Threshold: An Important Predictor of Hearing Preservation in Cochlear Implantation With Lateral Wall Electrodes.

OBJECTIVE: To evaluate the outcomes and association factors of long-term hearing preservation (HP) in cochlear implantation with lateral wall (LW) electrode arrays. STUDY DESIGN: Retrospective case review. SETTING: Tertiary academic center. PATIENTS: Thirty-four consecutive ears from 32 patients with a ≤ 80 dB HL preoperative low-frequency pure-tone average of 250 to 500 Hz were included. INTERVENTION: Cochlear implantation with LW electrode arrays and the intention of achieving HP. MAIN OUTCOME MEASURES: HP classifications according to the HEARRING group and functional HP methods (≤ 85 dB HL of pure-tone threshold at 250 Hz) at 1 year postoperatively. RESULTS: Based on the HEARRING classification, complete, partial, and minimal HP was achieved in 7 ears (21%), 12 ears (35%), and 4 ears (12%), respectively. Under functional classification, 14 ears (41%) exhibited functional hearing after surgery. The average threshold shift was 17.1 dB HL (SD = 16.9 dB HL). Among various clinical features, a ≤ 60 dB HL preimplant pure-tone threshold of 250 Hz was associated with HP outcome in both classifications (OR = 12.95, 95% CI = 1.29-130.01, p = 0.029 in HEARRING classification; OR = 14.36, 95% CI = 1.07-191.40, p = 0.044 in functional classification). The following parameters were not associated with HP (p > 0.05): patient demographics, surgical aspects (insertion route and depth), electrode array size, scalar electrode position, and presence of comorbidity. CONCLUSION: With LW electrode arrays, HP was achieved in 68% of HEARRRING group patients and 41% of functional classification patients. A ≤ 60 dB HL preimplant pure-tone threshold of 250 Hz was significantly associated with an increased rate of long-term HP.

Neuropilin-2 promotes growth and progression of papillary thyroid cancer cells.

OBJECTIVE: Neuropilin-2 (NRP2) is a coreceptor of vascular endothelial growth factor-C/D (VEGF-C/D) and plays the important role in the development of lymphatic endothelial cells, as well as neuronal development. NRP2 is known to affect aggressiveness by increasing expression in various human cancers, but the role of NRP2 in thyroid cancer is not fully understood. The purpose of this study was to investigate the NRP2 expression and its role in regulating the tumor aggressiveness in the papillary thyroid carcinoma (PTC). METHODS: The NRP2 expression and its clinicopathologic correlation to PTC was determined using the data from the 262 PTC patients at a tertiary referral medical center and The Cancer Genome Atlas (TCGA) database. The potential role of NRP2 in modulating tumor growth, invasion, and metastasis in PTC was examined by using small interfering RNA (siRNA)-mediated knockdown of NRP2. RESULTS: High expression of NRP2 was significantly associated with capsular invasion, lymphovascular invasion, lymph node metastasis, 5 or more metastatic lymph nodes, and recurrence in PTC patients. In TCGA data, the higher NRP2 expression group was significantly associated with extrathyroid extension, lymph node metastasis, and BRAFV600E mutation. The siRNA mediated knockdown of NRP2 in the PTC cells reduced the cell proliferation, migration and invasion. We also have confirmed that NRP2 knockdown suppressed epithelial-mesenchymal transition (EMT) by regulating AKT and ERK phosphorylation signaling pathways. CONCLUSION: Our results suggest that NRP2 regulates tumor progression in PTC and may act as a predictive factor for aggressiveness of PTC.

Clinical characteristics and associated factors of canal switch in benign paroxysmal positional vertigo.

BACKGROUND: Canal switch benign paroxysmal positional vertigo (CS-BPPV) is a transition of BPPV involving one canal to another canal during or after canalith repositioning procedures (CRP). OBJECTIVE: To investigate the clinical characteristics of CS-BPPV and its associated factors. METHODS: The data of 2,303 patients with BPPV involving the lateral canal (LC) or posterior canal (PC) were retrospectively analyzed. Demographics, etiologies, and various clinical parameters related to CRP were compared between patients with and without CS-BPPV. RESULTS: Sixty-eight (2.95%) patients exhibited CS-BPPV. For patients with CS-BPPV from the PC to the LC, as well as those with CS-BPPV from the LC to the PC, the CRP number for the original canal in CS-BPPV was significantly greater than in non-CS-BPPV (P = 0.002). More CRP cycles were required to treat CS-BPPV than non-CS-BPPV involving the same canal. Multivariate analysis showed that CS-BPPV from the LC to the PC was significantly associated with multiple CRP cycles and use of the Gufoni maneuver (P = 0.038 and P < 0.001, respectively). CONCLUSIONS: The use of multiple cycles of CRP and the Gufoni maneuver were significantly associated with the onset of CS-BPPV. Furthermore, more CRP cycles were needed for the treatment of CS-BPPV than for non-CS-BPPV involving the same canal.

New finding and optimal production of a novel extracellular alkaline lipase from Yarrowia lipolytica NRRL Y-2178.

Lipases are industrially useful versatile enzymes that catalyze numerous different reactions including hydrolysis of triglycerides, transesterification, and chiral synthesis of esters under natural conditions. Although lipases from various sources have been widely used in industrial applications, such as in food, chemical, pharmaceutical, and detergent industries, there are still substantial current interests in developing new microbial lipases, specifically those functioning in abnormal conditions. We screened 17 lipase-producing yeast strains, which were prescreened for substrate specificity of lipase from more than 500 yeast strains from the Agricultural Research Service Culture Collection (Peoria, IL, U.S.A.), and selected Yarrowia lipolytica NRRL Y-2178 as a best lipase producer. This report presents new finding and optimal production of a novel extracellular alkaline lipase from Y. lipolytica NRRL Y-2178. Optimal c ulture conditions f orlipase production by Y. lipolytica NRRL Y-2178 were 72 h incubation time, 27.5 degrees C, pH 9.0. Glycerol and glucose were efficiently used as the most efficient carbon sources, and a combination of yeast extract and peptone was a good nitrogen source for lipase production by Y. lipolytica NRRL Y-2178. These results suggested that Y. lipolytica NRRL Y-2178 showsgood industrial potential as a new alkaline lipase producer.

The effect of bio-converted polyunsaturated fatty acids on the oxidation of TAG containing highly unsaturated fatty acids.

Microbial modification of vegetable fatty acids can often lead to special changes in their structure and in biological function. A bacterial strain, Pseudomonas aeruginosa PR3, is known to carry out multiple hydroxylations on polyunsaturated fatty acids containing 1,4-cis, cis diene structural units, resulting in antibacterial activity. In this paper, in an effort to understand the overall mechanism involved in the varied biological functions of the complicated metabolites of bio-converted polyunsaturated fatty acids, we performed bioconversion of several polyunsaturated fatty acids using PR3, and determined their oxidative activities against fish oil. Bio-converted linoleic acid, eicosapentanoic acid, and docosahexanoic acid promoted effectively oxidation of fish oil. It is assumed that this oxidative effect could plausibly play an important role in the antimicrobial function of these bio-converted fatty acids.