Effect of a Combination of Lactiplantibacillus plantarum KC3 and Leonurus japonicus Extracts in Respiratory Discomfort: A Randomized, Double-Blind, Placebo-Controlled Trial.

The increased global prevalence of chronic respiratory diseases in recent years has caused a substantial public health burden. Lactiplantibacillus plantarum KC3 and Leonurus japonicus Houtt. (LJH) extracts can alleviate respiratory symptoms and improve lung function in vitro and in vivo. However, the clinical efficacy and safety profile of this combination in patients with respiratory diseases remain unclear. Therefore, this multicenter, randomized, double-blind, placebo-controlled clinical trial aimed to evaluate the efficacy and safety of L. plantarum KC3 and LJH extracts in adults with respiratory discomfort. This mixture was termed 'CKDB-315'. Participants, randomly assigned to the CKDB-315 or placebo groups, were treated for 12 weeks. Assessments included the St. George's Respiratory Questionnaire (SGRQ) and the Chronic Obstructive Pulmonary Disease Assessment Test (CAT). The CKDB-315 group showed considerably improved SGRQ and CAT scores compared with the placebo group. Secondary outcomes, including dyspnea, pulmonary function, total antioxidant status, and inflammatory cytokine levels, were consistent with the primary outcomes. Exploratory analyses of the gut microbiota and short-chain fatty acid contents revealed the potential mechanisms underlying the effects of CKDB-315. Finally, safety analysis indicated that CKDB-315 was well tolerated and caused few adverse events. Our findings indicate that CKDB-315 is a promising therapeutic option for respiratory discomfort in adults.

Precision Estimation of Crop Coefficient for Maize Cultivation Using High-Resolution Satellite Imagery to Enhance Evapotranspiration Assessment in Agriculture.

The estimation of crop evapotranspiration (ETc) is crucial for irrigation water management, especially in arid regions. This can be particularly relevant in the Po Valley (Italy), where arable lands suffer from drought damages on an annual basis, causing drastic crop yield losses. This study presents a novel approach for vegetation-based estimation of crop evapotranspiration (ETc) for maize. Three years of high-resolution multispectral satellite (Sentinel-2)-based Normalized Difference Vegetation Index (NDVI), Normalized Difference Water Index (NDWI), Normalized Difference Red Edge Index (NDRE), and Leaf Area Index (LAI) time series data were used to derive crop coefficients of maize in nine plots at the Acqua Campus experimental farm of Irrigation Consortium for the Emilia Romagna Canal (CER), Italy. Since certain vegetation indices (VIs) (such as NDVI) have an exponential nature compared to the other indices, both linear and power regression models were evaluated to estimate the crop coefficient (Kc). In the context of linear regression, the correlations between Food and Agriculture Organization (FAO)-based Kc and NDWI, NDRE, NDVI, and LAI-based Kc were 0.833, 0.870, 0.886, and 0.771, respectively. Strong correlation values in the case of power regression (NDWI: 0.876, NDRE: 0.872, NDVI: 0.888, LAI: 0.746) indicated an alternative approach to provide crop coefficients for the vegetation period. The VI-based ETc values were calculated using reference evapotranspiration (ET0) and VI-based Kc. The weather station data of CER were used to calculate ET0 based on Penman-Monteith estimation. Out of the Vis, NDWI and NDVI-based ETc performed the best both in the cases of linear (NDWI RMSE: 0.43 ± 0.12; NDVI RMSE: 0.43 ± 0.095) and power (NDWI RMSE: 0.44 ± 0.116; NDVI RMSE: 0.44 ± 0.103) approaches. The findings affirm the efficacy of the developed methodology in accurately assessing the evapotranspiration rate. Consequently, it offers a more refined temporal estimation of water requirements for maize cultivation in the region.

Prevalence of Keratoconus in the Young Eastern Population of Saudi Arabia.

BACKGROUND: Keratoconus (KCN) is a progressive corneal ectasia that manifests at a young age and significantly impacts vision and quality of life. Early diagnosis allows for effective treatment with corneal collagen crosslinking, yet there is a lack of screening methods. This research aims to screen adolescents and young adults for this sight-threatening disease using quick corneal tomography mapping. METHODS: This prospective cross-sectional study is being conducted at Johns Hopkins Aramco Healthcare in Saudi Arabia, focusing on subjects aged 13-23. We are presenting the data from our study as internal pilot study data. Bilateral corneal imaging with Pentacam HR (Oculus, Wetzlar, Germany), utilizing Scheimpflug corneal tomography, was performed. Historical data on allergies, eye rubbing, KCN, family history, previous eye surgery, and contact lens use were collected. The Belin Ambrosio Enhanced Ectasia Display total D value served as an objective criterion for suspect KCN (SKCN) diagnosis. RESULTS: In this study with 110 participants, KCN was identified in 2.75% of participants and SKCN in 11.93%. Systemic allergies or eczema were reported by 2.80%, with no cases in the KCN or SKCN groups. Eye rubbing behavior was observed in 5.50%, with the highest prevalence (33.30%) in the KCN group. A family history of KCN was found in 21.10%, with SKCN having the highest prevalence (30.80%). CONCLUSION: This restricted population study reveals a significant KCN rate of 2.75%. The condition, easily detected and treatable with corneal collagen crosslinking, highlights the need for larger population studies to determine the disease's true prevalence. Efficient screening programs tailored to regional data are essential for early detection and intervention.

Correlation Between the Cost and Safety of Corneal Graft Types.

Background Corneal diseases are the fourth most common cause of blindness worldwide. In the majority of these diseases, vision reduction is reversible and can be restored to a large extent by replacing the cornea through specific surgery and, in particular, transplantation. In Greece, due to a lack of organized eye banks as well as donors, the grafts intended for corneal transplantation usually come from eye banks abroad. This study focuses on the dynamics of cost versus value in the decision-making process for the procurement of corneal grafts, ultimately investigating the safety that the procured grafts provide to patients. Methodology A total of 267 patients with severe vision problems who underwent 301 corneal and amniotic membrane transplants from years 2020 to 2023 at the Transplant Unit of the Athens General Hospital "Georgios Gennimatas" were included in this study. All patients who were deemed appropriate to undergo corneal transplant operations, the diagnosis that led to the specific surgery, and other relevant data were recorded and evaluated. Results There was no significant difference in the ratio between males and females (51.3% male and 48.7% female). The mean age of the patients was 66.5 years (SD = 13.7 years). Graft problems were faced by 13.9% of the patients, with the amniotic membrane by 1.5% (in the total number of surgical operations) and both eyes by 4.5% of patients. The majority of the patients had undergone only one surgery (88.8%). Reoperation was needed in 14% of the cases, and 7.6% of the cases were surgeries that occurred due to graft rejection or non-functioning grafts from surgeries performed at another hospital or clinic. In the majority of surgeries (60.8%), a Descemet's stripping automated endothelial keratoplasty graft was used. The mean cost was 3,167 euro (SD = 960.3 euro). Furthermore, in 35.8% of the surgeries, the graft was preserved with amphotericin. Conclusions The present study draws useful conclusions about the effectiveness of surgical interventions through the correlation of cost and safety of the grafts that are approved and finally used in corneal transplants, as well as the submission of proposals to improve the procedures and lead to patient benefits.

Upregulated Matrisomal Proteins and Extracellular Matrix Mechanosignaling Underlie Obesity-Associated Promotion of Pancreatic Ductal Adenocarcinoma.

Diet-induced obesity (DIO) promotes pancreatic ductal adenocarcinoma (PDAC) in mice expressing KRasG12D in the pancreas (KC mice), but the precise mechanisms remain unclear. Here, we performed multiplex quantitative proteomic and phosphoproteomic analysis by liquid chromatography-tandem mass spectrometry and further bioinformatic and spatial analysis of pancreas tissues from control-fed versus DIO KC mice after 3, 6, and 9 months. Normal pancreatic parenchyma and associated proteins were steadily eliminated and the novel proteins, phosphoproteins, and signaling pathways associated with PDAC tumorigenesis increased until 6 months, when most males exhibited cancer, but females did not. Differentially expressed proteins and phosphoproteins induced by DIO revealed the crucial functional role of matrisomal proteins, which implies the roles of upstream regulation by TGFß, extracellular matrix-receptor signaling to downstream PI3K-Akt-mTOR-, MAPK-, and Yap/Taz activation, and crucial effects in the tumor microenvironment such as metabolic alterations and signaling crosstalk between immune cells, cancer-associated fibroblasts (CAFs), and tumor cells. Staining tissues from KC mice localized the expression of several prognostic PDAC biomarkers and elucidated tumorigenic features, such as robust macrophage infiltration, acinar-ductal metaplasia, mucinous PanIN, distinct nonmucinous atypical flat lesions (AFLs) surrounded by smooth muscle actin-positive CAFs, invasive tumors with epithelial-mesenchymal transition arising close to AFLs, and expanding deserted areas by 9 months. We next used Nanostring GeoMX to characterize the early spatial distribution of specific immune cell subtypes in distinct normal, stromal, and PanIN areas. Taken together, these data richly contextualize DIO promotion of Kras-driven PDAC tumorigenesis and provide many novel insights into the signaling pathways and processes involved.

Genetic investigation in a four-generation Chinese family with congenital fibrosis of extraocular muscles and keratoconus.

Here, we have reported the genetic and clinical characteristics of four generations of a family patient from China with congenital fibrosis of extraocular muscles 1 (CFEOM1) and keratoconus (KC). The history of diseases, clinical observations, and blood samples of all family members were collected. A total of 100 healthy participants were recruited as normal controls. The whole exome sequencing of the genomic DNA and polymerase chain reaction were performed on samples obtained from the controls and their family members to verify the gene variants. The functional analyses of the variants were performed by using different software. Two single nucleotide polymorphisms were detected in the proband and other patients in his families, including a heterozygous missense variation, g.39726207C > T (c.2860C > T, p.R954W, rs121912585), in the third highly conserved coiled-coil domain of KIF21A, and a heterozygous missense variant, g.30664732A > C (c.136A > C, p.S46R, rs200111443) in TGFBR2. The variant p.R954W in KIF21A was predicted to be pathogenic using software, whereas p.S46R in TGFBR2 was predicted to be of uncertain significance (VUS). Thus, KC might have occurred in the proband and his daughter because of a combination of genetic mutations and involuntary eye rubbing induced by CFEOM1. This is the first case of concomitant KC in a family having CFEOM1. Thus, the study provides new information about patients with KC having CFEOM1. Furthermore, the study suggests that attention should be paid to the early detection and diagnosis of KC in patients with CFEOM1.

Transcription factor expression landscape in Drosophila embryonic cell lines.

BACKGROUND: Transcription factor (TF) proteins are a key component of the gene regulatory networks that control cellular fates and function. TFs bind DNA regulatory elements in a sequence-specific manner and modulate target gene expression through combinatorial interactions with each other, cofactors, and chromatin-modifying proteins. Large-scale studies over the last two decades have helped shed light on the complex network of TFs that regulate development in Drosophila melanogaster. RESULTS: Here, we present a detailed characterization of expression of all known and predicted Drosophila TFs in two well-established embryonic cell lines, Kc167 and S2 cells. Using deep coverage RNA sequencing approaches we investigate the transcriptional profile of all 707 TF coding genes in both cell types. Only 103 TFs have no detectable expression in either cell line and 493 TFs have a read count of 5 or greater in at least one of the cell lines. The 493 TFs belong to 54 different DNA-binding domain families, with significant enrichment of those in the zf-C2H2 family. We identified 123 differentially expressed genes, with 57 expressed at significantly higher levels in Kc167 cells than S2 cells, and 66 expressed at significantly lower levels in Kc167 cells than S2 cells. Network mapping reveals that many of these TFs are crucial components of regulatory networks involved in cell proliferation, cell-cell signaling pathways, and eye development. CONCLUSIONS: We produced a reference TF coding gene expression dataset in the extensively studied Drosophila Kc167 and S2 embryonic cell lines, and gained insight into the TF regulatory networks that control the activity of these cells.

Awareness of Keratoconus and Its Relationship With Eye Rubbing Among the Population of the Eastern Province of Saudi Arabia.

BACKGROUND: Keratoconus is an eye condition where the cornea, the clear, dome-shaped front part of the eye, becomes thinner and gradually bulges outward into a cone shape. Keratoconus usually affects both eyes. The exact cause of keratoconus is unknown, but it is believed to involve a combination of genetic and environmental factors. AIM: This study aims to assess the awareness level of keratoconus and its relation to eye rubbing among the population in the Eastern Province of Saudi Arabia. METHODS: A cross-sectional study was conducted in the Eastern Province of Saudi Arabia, involving all eligible participants via an online pre-designed questionnaire from March to June 2023. The data were collected through social media. RESULTS: A total of 388 eligible participants completed the study questionnaire. Participants' ages ranged from 19 to 60 years, with a mean age of 26.2 ± 13.9 years. There were 265 (68.3%) female participants. Regarding public awareness about keratoconus among participants in the Eastern Province of Saudi Arabia, our research revealed a total of 101 (26%) participants had an overall good awareness of keratoconus, while 287 (74%) had a poor awareness level. The most reported source of information included scientific lectures (21.6%). Precisely, 316 (81.4%) of the research subjects rub their eyes primarily because of allergies (7%), strain headaches (25%), and itching (66.5%). CONCLUSION: Most of the study participants have poor awareness about keratoconus and its relation to eye rubbing. Health education programs for the population should be conducted to enhance public awareness about keratoconus.

Investigation of How Corneal Densitometry Artefacts Affect the Imaging of Normal and Keratoconic Corneas.

PURPOSE: To investigate corneal densitometry artefacts found in Pentacam Scheimpflug scans and their potential effect on assessing keratoconic (KC) corneas compared to normal (N) corneas. METHODS: The current study utilises Pentacam data of 458 N eyes, aged 35.6 ± 15.8 (range 10-87), referred to as the "N group", and 314 KC eyes, aged 31.6 ± 10.8 (range 10-72), referred to as the "KC group", where densitometry data were extracted and analysed via a custom-built MATLAB code. Radial summations of the densitometry were calculated at diameters ranging from 0.5 mm to 5.0 mm. The minimum normalised radial summation of densitometry (NRSD) value and angle were determined at each diameter and then linked. KC cone locations and areas of pathology were determined, and a comparison between N and KC groups was carried out both within the averaged area of pathology and over the corneal surface. RESULTS: Joining minimum NRSD trajectory points marked a clear distortion line pointing to the nasal-superior direction at 65° from the nasal meridian. The findings were found to be independent of eye laterality or ocular condition. Consistency was detected in the right and left eyes among both the N and KC groups. The location of the KC cone centre and the area of pathology were determined, and the densitometry output was compared both within the area of pathology and over the whole cornea. When the average densitometry was compared between N and KC eyes within the KC area of pathology, the N group recorded a 16.37 ± 3.15 normalised grey-scale unit (NGSU), and the KC group recorded 17.74 ± 3.4 NGSU (p = 0.0001). However, when the whole cornea was considered, the N group recorded 16.71 ± 5.5 NGSU, and the KC group recorded 15.72 ± 3.98 NGSU (p = 0.0467). A weak correlation was found between the Bad D index and NGSU when the whole measured cornea was considered (R = -0.01); however, a better correlation was recorded within the KC area of pathology (R = 0.21). CONCLUSIONS: Nasal-superior artefacts are observed in the densitometry Pentacam maps, and analysis shows no significant differences in their appearance between N or KC corneas. When analysing KC corneas, it was found that the cone positions are mostly on the temporal-inferior side of the cornea, opposite to the densitometry artefact NRSD trajectory. The analysis suggests that the corneal densitometry artefacts do not interfere with the KC area of pathology as it reaches its extreme in the opposite direction; therefore, weighting the densitometry map to increase the contribution of the inferior-temporal cornea and decreasing that of the superior-nasal area would improve the classification or identification of KC if densitometry is to be used as a KC metric.

Advances in the Treatment of Keratoconus: Epithelial-On (EPI-On) Corneal-Collagen Cross-Linking (CXL) and CXL-Plus Procedures.

Keratoconus (KC) incidence is on the increase. The advent of corneal-collagen cross-linking (CXL) has revolutionized the management of KC. This systematic review looks at the efficacy and complications of two novel treatments within CXL: Epithelial-On (Epi-On) and CXL-plus procedures. Two separate literature searches were carried out up until July 1, 2021. Articles only published in the last two years were included to ensure that only the most recent articles were reviewed. A total of 15 articles were selected for this review. There were varied results regarding the efficacy of Epi-On. No significant difference was found between Epi-On and standard Epithelial-Off (Epi-Off) CXL. However, it was found that Epi-On was inferior to standard CXL in terms of reducing KMAX. There was a higher risk of progression in patients treated with Epi-On CXL, with an increased rate of patients requiring re-treatment due to the advancement of their KC. While some studies report CXL-plus procedures demonstrate long-term efficacy and safety, a considerable number of studies advise caution, reporting a significant deterioration in corrected distance visual acuity (CDVA). Consequently, a question persists regarding the safest and most efficacious approach, given the lack of robust large randomized controlled trials (RCTs) in the current literature.

Protective effect of the mixture of Lactiplantibacillus plantarum KC3 and Leonurus Japonicas Houtt extract on respiratory disorders.

Air pollutants, such as particulate matter (PM) and diesel exhaust particles (DEP), are associated with respiratory diseases. Therefore, preventive and therapeutic strategies against PM-and DEP (PM10D)-induced respiratory diseases are needed. Herein, we evaluate the protective effects of a mixture of Lactiplantibacillus plantarum KC3 and Leonurus Japonicas Houtt (LJH) extract against airway inflammation associated with exposure to PM10D. To determine the anti-inflammatory effects of the LJH extract, reactive oxygen species (ROS) production and the expression of inflammatory pathways were determined in PM10-induced MH-S cells. For the respiratory protective effects, BALB/c mice were exposed to PM10D via intranasal injection, and a mixture of L. plantarum KC3 and LJH extract was administered orally for 12 days. LJH extract inhibited ROS production and the phosphorylation of downstream factors of NF-κB in PM10-stimulated MH-S cells. The mixture of L. plantarum KC3 and LJH repressed the infiltration of neutrophils, reduced the immune cells number, and suppressed the proinflammatory mediators and cyclooxygenase (COX)-2 expressions in PM10D-induced airway inflammation with reduced phosphorylation of downstream factors of NF-κB. In addition, these effects were not observed in an alveolar macrophage depleted PM10D-induced mouse model using clodronate liposomes. The extract mixture also regulated gut microbiota in feces and upregulated the mRNA expression of Foxp3, transforming growth factor (TGF)-ß1, and interleukin (IL)-10 in the colon. The L. plantarum KC3 and LJH extract mixture may inhibit alveolar macrophage- and neutrophil-mediated inflammatory responses and regulate gut microbiota and immune response in PM10D-induced airway inflammation, suggesting it is a potential remedy to prevent and cure airway inflammation and respiratory disorders.

Targeting autophagy and beyond: Deconvoluting the complexity of Beclin-1 from biological function to cancer therapy.

Beclin-1 is the firstly-identified mammalian protein of the autophagy machinery, which functions as a molecular scaffold for the assembly of PI3KC3 (class III phosphatidylinositol 3 kinase) complex, thus controlling autophagy induction and other cellular trafficking events. Notably, there is mounting evidence establishing the implications of Beclin-1 in diverse tumorigenesis processes, including tumor suppression and progression as well as resistance to cancer therapeutics and CSC (cancer stem-like cell) maintenance. More importantly, Beclin-1 has been confirmed as a potential target for the treatment of multiple cancers. In this review, we provide a comprehensive survey of the structure, functions, and regulations of Beclin-1, and we discuss recent advances in understanding the controversial roles of Beclin-1 in oncology. Moreover, we focus on summarizing the targeted Beclin-1-regulating strategies in cancer therapy, providing novel insights into a promising strategy for regulating Beclin-1 to improve cancer therapeutics in the future.

Bubbling With Confidence: A Study of Numerous Deep Anterior Lamellar Keratoplasty Cases Using Anwar's Big Bubble Technique.

PURPOSE: This study aims to assess the postoperative results, variability, and complications of a hundred deep anterior lamellar keratoplasty cases. Study design and duration: This is an observational study. The study was conducted at Pak International Medical College (PIMC) for a duration of four years (January 2019-January 2023). METHODOLOGY: Our study collected information on a hundred cases of deep anterior lamellar keratoplasty (DALK) utilizing Anwar's big bubble technique, consisting of patients with keratoconus, superficial corneal scars, and macular dystrophy. Consenting patients had their pre and postoperative visual acuities and keratometry readings recorded. Overall success and complications were recorded and compared with the present literature. RESULTS: Big bubble formation was achieved in 87% (n = 87) eyes and not achieved in 13% (n=13). There was a significant reduction in keratometry values after the procedures as well as improved vision in all patients, with 84% reporting significant improvement. Descemet membrane exposure was achieved in 91% (n=91). Complications included the failure of Anwar's big bubble formation in 13% (n=13) patients and the failure to expose Descemet's in nine patients (9%). CONCLUSION: DALK using the big bubble technique is a safe and effective procedure in patients with corneal diseases who have a healthy Descemet membrane and endothelium.

Building Automation Pipeline for Diagnostic Classification of Sporadic Odontogenic Keratocysts and Non-Keratocysts Using Whole-Slide Images.

The microscopic diagnostic differentiation of odontogenic cysts from other cysts is intricate and may cause perplexity for both clinicians and pathologists. Of particular interest is the odontogenic keratocyst (OKC), a developmental cyst with unique histopathological and clinical characteristics. Nevertheless, what distinguishes this cyst is its aggressive nature and high tendency for recurrence. Clinicians encounter challenges in dealing with this frequently encountered jaw lesion, as there is no consensus on surgical treatment. Therefore, the accurate and early diagnosis of such cysts will benefit clinicians in terms of treatment management and spare subjects from the mental agony of suffering from aggressive OKCs, which impact their quality of life. The objective of this research is to develop an automated OKC diagnostic system that can function as a decision support tool for pathologists, whether they are working locally or remotely. This system will provide them with additional data and insights to enhance their decision-making abilities. This research aims to provide an automation pipeline to classify whole-slide images of OKCs and non-keratocysts (non-KCs: dentigerous and radicular cysts). OKC diagnosis and prognosis using the histopathological analysis of tissues using whole-slide images (WSIs) with a deep-learning approach is an emerging research area. WSIs have the unique advantage of magnifying tissues with high resolution without losing information. The contribution of this research is a novel, deep-learning-based, and efficient algorithm that reduces the trainable parameters and, in turn, the memory footprint. This is achieved using principal component analysis (PCA) and the ReliefF feature selection algorithm (ReliefF) in a convolutional neural network (CNN) named P-C-ReliefF. The proposed model reduces the trainable parameters compared to standard CNN, achieving 97% classification accuracy.

Prevalence of Psychiatric Illness Among Keratoconus Patients.

Background  Keratoconus is a chronic corneal disorder characterized by progressive thinning of the cornea resulting in visual deterioration. Keratoconus patients have a higher rate of psychiatric morbidities. Therefore, this study will assess the prevalence of psychiatric illness among keratoconus patients. Methods We conducted a quantitative retrospective study in three centers across Saudi Arabia from June 2016 to February 2023. We included all patients previously diagnosed with keratoconus and any psychiatric illness. We excluded patients with other ocular diseases in addition to keratoconus. After we extracted the data, we coded and analyzed them using IBM SPSS Statistics for Windows, Version 21.0 (2012; IBM Corp., Armonk, New York, United States) and GraphPad Prism Version 8.4.2 (Dotmatics, Boston, Massachusetts, United States). Results The study included 57 keratoconus patients, with the majority being Saudi (96.5%), female (2:1), in the age group of 31-40 years (40.4%), and with a high-school level of education. The majority of patients were also obese (42.4%) and non-smokers (82%). Most patients had comorbid medical disorders. Of the 57 patients, 63.2% had anxiety disorders, 56.1% had depression, 10.5% had schizophrenia, and 1.8% had bipolar disorder. Patients with only a primary-school level of education were significantly more likely to be diagnosed with bipolar disorder, whereas those who were single were significantly more likely to be diagnosed with schizophrenia. Corneal transplant was also significantly associated with schizophrenia. Most patients (51%) were diagnosed with a psychiatric illness before keratoconus was diagnosed. Conclusion Among the keratoconus patients, single relationship status, primary-school level of education, and comorbidities were significantly associated with schizophrenia, bipolar disorder, and anxiety disorder, respectively. In addition, corneal transplant was significantly associated with schizophrenia. Lastly, the occurrence of psychiatric illness before keratoconus diagnosis was observed in the majority of patients.

Corynoxine B targets at HMGB1/2 to enhance autophagy for α-synuclein clearance in fly and rodent models of Parkinson's disease.

Parkinson's disease (PD) is the most common neurodegenerative movement disease. It is featured by abnormal alpha-synuclein (α-syn) aggregation in dopaminergic neurons in the substantia nigra. Macroautophagy (autophagy) is an evolutionarily conserved cellular process for degradation of cellular contents, including protein aggregates, to maintain cellular homeostasis. Corynoxine B (Cory B), a natural alkaloid isolated from Uncaria rhynchophylla (Miq.) Jacks., has been reported to promote the clearance of α-syn in cell models by inducing autophagy. However, the molecular mechanism by which Cory B induces autophagy is not known, and the α-syn-lowering activity of Cory B has not been verified in animal models. Here, we report that Cory B enhanced the activity of Beclin 1/VPS34 complex and increased autophagy by promoting the interaction between Beclin 1 and HMGB1/2. Depletion of HMGB1/2 impaired Cory B-induced autophagy. We showed for the first time that, similar to HMGB1, HMGB2 is also required for autophagy and depletion of HMGB2 decreased autophagy levels and phosphatidylinositol 3-kinase III activity both under basal and stimulated conditions. By applying cellular thermal shift assay, surface plasmon resonance, and molecular docking, we confirmed that Cory B directly binds to HMGB1/2 near the C106 site. Furthermore, in vivo studies with a wild-type α-syn transgenic drosophila model of PD and an A53T α-syn transgenic mouse model of PD, Cory B enhanced autophagy, promoted α-syn clearance and improved behavioral abnormalities. Taken together, the results of this study reveal that Cory B enhances phosphatidylinositol 3-kinase III activity/autophagy by binding to HMGB1/2 and that this enhancement is neuroprotective against PD.

Small cytosolic double-stranded DNA represses cyclic GMP-AMP synthase activation and induces autophagy.

The cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway is a major mediator of inflammation following stimulation with >45 bp double-stranded DNA (dsDNA). Herein, we identify a class of ∼20-40 bp small cytosolic dsDNA (scDNA) molecules that compete with long dsDNA (200-1,500 bp herring testis [HT]-DNA) for binding to cGAS, thus repressing HT-DNA-induced cGAS activation. The scDNA promotes cGAS and Beclin-1 interaction, releasing Rubicon, a negative regulator of phosphatidylinositol 3-kinase class III (PI3KC3), from the Beclin-1-PI3KC3 complex. This leads to PI3KC3 activation and induces autophagy, causing degradation of STING and long cytosolic dsDNA. Moreover, DNA damage decreases, and autophagy inducers increase scDNA levels. scDNA transfection and treatment with autophagy inducers attenuate DNA damage-induced cGAS activation. Thus, scDNA molecules serve as effective brakes for cGAS activation, preventing excessive inflammatory cytokine production following DNA damage. Our findings may have therapeutic implications for cytosolic DNA-associated inflammatory diseases.

Potential Therapeutic Functions of PU-91 and Quercetin in Personalized Cybrids Derived from Patients with Age-Related Macular Degeneration, Keratoconus, and Glaucoma.

The aim of this study is to investigate the therapeutic potential of higher doses of PU-91, quercetin, or in combination on transmitochondrial cybrid cell lines with various mtDNA haplogroups derived from patients with age-related macular degeneration (AMD), glaucoma (Glc), keratoconus (KC), and normal (NL) individuals. Cybrids were treated with PU-91 (P) (200 µM) alone, quercetin (Q) (20 µM) alone, or a combination of PU-91 and quercetin (P+Q) for 48 h. Cellular metabolism and the intracellular levels of reactive oxygen species (ROS) were measured by MTT and H2DCFDA assays, respectively. Quantitative real-time PCR was performed to measure the expression levels of genes associated with mitochondrial biogenesis, antioxidant enzymes, inflammation, apoptosis, and senescence pathways. PU-91(P) (i) improves cellular metabolism in AMD cybrids, (ii) decreases ROS production in AMD cybrids, and (iii) downregulates the expression of LMNB1 in AMD cybrids. Combination treatment of PU-91 plus quercetin (P+Q) (i) improves cellular metabolism in AMD, (ii) induces higher expression levels of TFAM, SOD2, IL6, and BAX in AMD cybrids, and (iii) upregulates CDKN1A genes expression in all disease cybrids. Our study demonstrated that the P+Q combination improves cellular metabolism and mitochondrial biogenesis in AMD cybrids, but senescence is greatly exacerbated in all cybrids regardless of disease type by the P+Q combined treatment.

Enhanced osteogenesis of human urine-derived stem cells by direct delivery of 30Kc19α-Lin28A protein.

Urine-derived stem cells (USCs) are a promising source for regenerative medicine because of their advantages such as easy and non-invasive collection from the human body, stable expansion, and the potential to differentiate into multiple lineages, including osteoblasts. In this study, we propose a strategy to enhance the osteogenic potential of human USCs using Lin28A, a transcription factor that inhibits let-7 miRNA processing. To address concerns regarding the safety of foreign gene integration and potential risk of tumorigenicity, we intracellularly delivered Lin28A as a recombinant protein fused with a cell-penetrating and protein-stabilizing protein, 30Kc19α. 30Kc19α-Lin28A fusion protein exhibited improved thermal stability and was delivered into USCs without significant cytotoxicity. 30Kc19α-Lin28A treatment elevated calcium deposition and upregulated several osteoblast-specific gene expressions in USCs derived from multiple donors. Our results indicate that intracellularly delivered 30Kc19α-Lin28A enhances the osteoblastic differentiation of human USCs by affecting the transcriptional regulatory network involved in metabolic reprogramming and stem cell potency. Therefore, 30Kc19α-Lin28A may provide a technical advancement toward developing clinically feasible strategies for bone regeneration.

Diffractive Trifocal Intraocular Lens Implantation in Stable Subclinical and Forme Fruste Keratoconus: A Case Series.

In this retrospective case series, we examine the outcomes of diffractive trifocal intraocular lens implantation after cataract surgery, in patients with subclinical and forme fruste keratoconus. Eight eyes of four patients (aged between 47 and 64) were included in the study and underwent phacoemulsification with implantation of AT LISA tri 839MP or AT LISA tri-toric 939MP intraocular lenses (Carl Zeiss Meditec AG, Jena, Germany). Post-operative evaluation included a visual acuity test at three distances (6m, 80cm, and 40cm), a visual acuity test at three low contrast levels (25%, 12.5%, and 6%), and a questionnaire about the patients' experience with photic phenomena and overall satisfaction with the achieved quality of vision. Our results show that spectacle freedom was achieved in all cases with a high satisfaction rate among participants. We hope our results would encourage surgeons to offer this technology to carefully selected candidates with stable subclinical and forme fruste keratoconus undergoing cataract surgery, giving them the possibility of achieving spectacle independence.