Skin Fibroblasts Research Articles

Species differences in glycerol-3-phosphate metabolism reveals trade-offs between metabolic adaptations and cell proliferation

The temperate climate-adapted brown hare (Lepus europaeus) and the cold-adapted mountain hare (Lepus timidus) are closely related and interfertile species. However, their skin fibroblasts display distinct gene expression profiles related to fundamental cellular processes. This indicates important metabolic divergence between the two species. Through targeted metabolomics and metabolite tracing, we identified species-specific variations in glycerol 3-phosphate (G3P) metabolism. G3P is a key metabolite of the G3P shuttle, which transfers reducing equivalents from cytosolic NADH to the mitochondrial electron transport chain (ETC), consequently regulating glycolysis, lipid metabolism, and mitochondrial bioenergetics. Alterations in G3P metabolism have been implicated in multiple human pathologies including cancer and diabetes. We observed that mountain hare mitochondria exhibit elevated G3P shuttle activity, alongside increased membrane potential and decreased mitochondrial temperature. Silencing mitochondrial G3P dehydrogenase (GPD2), which couples the conversion of G3P to the ETC, uncovered its species-specific role in controlling mitochondrial membrane potential and highlighted its involvement in skin fibroblast thermogenesis. Unexpectedly, GPD2 silencing enhanced wound healing and cell proliferation rates in a species-specific manner. Our study underscores the pivotal role of the G3P shuttle in mediating physiological, bioenergetic, and metabolic divergence between these hare species.

Evaluation of the cytotoxic activity of essential oils from Malaysian herbal plants against A375 and A431 skin cancer cell lines

Skin cancer incidences is rising due to prolonged harmful exposure to UV rays. Current therapies can adversely affect healthy cells. Conventional chemotherapy administered orally and intravenously poses risks of off targets, leading to severe and debilitating side effects. This study explored the anticancer potential of essential oils (EOs) extracted by hydrodistillation from seven local plant leaves (Piper betle, Persicaria minor, Cosmos caudatus, Citrus aurantifolia, Citrus hystrix, Citrus microcarpa, and Piper sarmentosum) on melanoma (A375), non-melanoma (A431), and fibroblasts skin cells (HFF1). Cytotoxicity assessment using MTT assay showed that all EOs tested had inhibitory effects of all EOs towards both cancer cell lines. Out of all seven plant EOs tested, P. sarmentosum EO displayed the most potent cytotoxicity at 80 ± 0.27 μg/mL with the extraction yield of 0.63%. Morphological changes in cellular size were observed after 4 h of EO treatment, leading to cell death coherently with declination of viability. This research signifies Malaysian plant’s role for topical application, as demonstrated by viability and morphology study.

New 5,6,7,8-Tetrahydro-isoquinolines Bearing 2-Nitrophenyl Group Targeting RET Enzyme: Synthesis, Anticancer Activity, Apoptotic Induction and Cell Cycle Arrest.

In this work, we synthesized new 5,6,7,8-tetrahydroisoquinolines and 6,7,8,9-tetrahydrothieno[2,3-c]isoquinolines derivatives and the structures of these new compounds were confirmed with different spectroscopic techniques. Furthermore, the anticancer activities of these compounds were assessed against eight tumor cell lines and one normal human skin fibroblast cell line (HSF). Subsequently, IC50 values of the synthesized compounds were determined for two specific cancer cell lines. Compound 3 exhibited the most potent antiproliferative activity against the HEPG2 cell line, whereas compound 9c demonstrated superior efficacy against the HCT116 cell line. Moreover, the mechanism of action for compound 3 on HEPG2 cells using flow cytometry and Annexin V-FITC apoptosis analysis was studied. Compound 3 caused cell cycle arrest at the G2/M with a 50-fold increase in apoptosis of the HEPG2 cell line. Finally, a molecular docking study was conducted to assess the inhibitory potential of compounds 3 and 7 against the RET enzyme. Results indicated that compounds 3 and 7 bind to the RET enzyme with binding energies of -5.2 and -5.6 kcal/mol, respectively. While these values suggest inhibitory activity, they are less potent than the standard inhibitor, alectinib, which exhibits a binding energy of -7.2 kcal/mol.

Antibacterial and anticancer activities of three novel lectin-conjugated chitosan nanoparticles

To the best of our knowledge, this is the first attempt to synthesize, characterize, and determine the antibacterial and anticancer effects of three novel conjugates of plant lectins: phytohemagglutinin lectin (PHA), soybean agglutinin (SBA), and peanut agglutinin (PNA) with chitosan nanoparticles (CHNPs). The lectin concentration within prepared conjugates was estimated using nannodrop, and the highest concentration was 0.96 mg/ml in PHA-CHNPs. SDS-PAGE showed the molecular weights of conjugates ranged from 26.9 to 63.9 kDa. UV spectrophotometer recorded the absorbance peaks of conjugates somewhere between 200 and 230 nm. Hemagglutination analysis verified the presence of actively binding lectins. The three conjugates showed strong antibacterial activity against Gram-positive and Gram-negative bacteria compared to pure lectins and chitosan nanoparticles. The highest inhibition zone was 55.67 ± 4.04, 38.67 ± 5.51, and 37.33 ± 2.52 for PHA-CHNPs against Enterococcus faecalis, Salmonella typhimurium, and Shigella sonnei, respectively, followed by 36.3 ± 0.15 for PNA-CHNPs against Staphylococcus aureus. The lowest MIC was 1.5 µg/ml for PHA-CHNPs against Enterococcus faecalis, followed by 12 µg/ml for PNA-CHNPs and SBA-CHNPs against Salmonella typhimurium and Enterococcus faecalis, respectively. TEM microphotographs show the conjugation pattern between lectins and chitosan nanoparticles and the morphological differences between control, treated bacteria, and cancer cells. Moreover, 100 μg/ml of PHA-CHNPs affect tongue carcinoma (HNO-97), colorectal cancer (HT-29), and human melanoma (A375) cancer cell lines, reducing cell viability by 38.78 ± 1.85%, 49.88 ± 1.11%, and 66.92 ± 3.60%, respectively. This study develops three innovative conjugates of lectin chitosan nanoparticles that need to be tested as potential antibacterial and anticancer agents for medical and cancer therapy applications.Key points• Lectin-conjugated chitosan nanoparticles exhibit antibacterial activity.• All conjugates are safe for oral epithelial cells and human skin fibroblasts.• The PHA-CHNP conjugates have anticancer activity against HNO-97, HT-29, and A375.

Human Skin Fibroblasts as an In Vitro Model Illustrating Changes in Collagen Levels and Skin Cell Migration Under the Influence of Selected Plant Hormones

Human skin fibroblasts are an excellent in vitro model for tracking the processes occurring in human skin and studying the potential impact of various biologically active substances on these processes. Two plant hormones, which are included in the cytokinins group—kinetin (K) and N-6-benzyladenine (BA)—have a positive effect on human skin. Therefore, an attempt was made to examine the effect they have on key skin functions, cell proliferation, and migration, as well as collagen synthesis in them. The effect of phytohormones was studied at selected concentrations for kinetin—10 μM and 1 μM—and for N-6-benzyladenine—1 μM and 0.1 μM. A wound-healing assay was used in order to analyze cell migration and proliferation. The content of total protein and collagen in cells and culture medium was determined. The obtained results confirm that the studied compounds induce cell migration and proliferation, as well as collagen biosynthesis. The positive effect of kinetin and N-6-benzyladenine on fibroblast metabolism that we have demonstrated allows us to indicate them as compounds with potentially therapeutic properties. Therefore, we conclude that they should be subjected to further molecular and in vivo studies focusing on pathologies connected with skin diseases and aging.

Intermittent episodes of acute severe encephalomyopathy and early death in two siblings caused by biallelic likely pathogenic variants in FASTKD2: Expanding phenotype and literature review.

Combined oxidative phosphorylation (OXPHOS) deficiency 44 (COXPD44; MIM# 618855) is caused by biallelic pathogenic variants in FAS-activated serine-threonine kinase domain 2 (FASTKD2) (MIM# 612322). COXPD44 is characterized by variable clinical features-developmental delay, chronic epileptic encephalopathy, seizure disorder/status epilepticus and cerebellar ataxia. We ascertained one sibwith episodic acute encephalomyopathy triggered by acute gastroenteritis and associated with haematological abnormalities, rhabdomyolysis leading to acute kidney injury, hypotensive shock leading to early death and a similarly affected sib with early death. Both siblings were normal neurologically in between the acute episodes. Whole exome sequencing (WES) was performed in the elder sibling. Mitochondrial respiratory chain enzyme activity assaywas performed in fibroblast cells and muscle tissue of the elder sibling. Also, Adenosine triphosphate (ATP) determination assay was done in fibroblast cells of the elder sibling. WES revealed compound heterozygous missense likely pathogenic variants in FASTKD2. Mitochondrial respiratory chain enzyme activity in muscle tissue showed reduced complex IV activity and ATP determination assay showed a reduction of ATP in skin fibroblasts. Herein, we report two siblings with novel clinical phenotype associated with COXPD44. Our report further validates the biallelic variants in FASTKD2 associated with the variable phenotypes and mitochondrial OXPHOS defect.

Synthesis of derived Cu(II) complex from Schiff base of Vitamin B6 and antihypertensive hydralazine: DNA interaction, biological application and molecular docking studies

A novel Schiff base ligand (E)-5-(hydroxymethyl)-2-methyl-4-((2-(phthalazin-1-yl)hydrazineylidene)methyl)pyridin-3-ol was synthesized through the condensation of antihypertensive hydralazine and Pyridoxal (Vitamin B6) ligands. This fabricated Schiff base (L) was a precursor for developing a unique Cu(II) complex. TheSchiff base ligand and Cu(II) complex structureswere characterized using various analytical tools, including IR, UV–Vis Spectroscopy and NMR. An octahedron geometry of this complex was proposed for the assigned molecular formula [Cu(L)2].3H2O. The molecular structures of theSchiff base ligand and its Cu(II) complex have been optimized using the B3LYP functional and the 6-311G++(d,p) basis set. Investigating the interaction of this prepared Cu(II) complex with calf thymus DNA (CT-DNA) molecules was operated using UV–Vis spectrophotometry, viscosity measurement, electrochemical studies, gel electrophoresis, and molecular docking. In vitro cytotoxicity was conducted using the MTT assay on four human tumor cell lines including hepatocellular carcinoma, estrogen-dependent breast cancer, melanoma, triple-negative breast cancer as well as healthy human skin fibroblasts.

Formulation, Evaluation, Factorial Optimization, and In-Silico Study of Eplerenone Loaded Pectin Nanoparticles: A New Approach to the Proliferation of Human Skin Fibroblasts for Wound Healing

Formulation, Evaluation, Factorial Optimization, and In-Silico Study of Eplerenone Loaded Pectin Nanoparticles: A New Approach to the Proliferation of Human Skin Fibroblasts for Wound Healing

An Integrated Approach to Develop Innovative, Sustainable, and Effective Cosmetic Ingredients: The Case Report of Fatty-Acids-Enriched Wild Strawberry Waste Extract

The sourcing of raw materials with low environmental impact, e.g., “upcycled” ingredients from short supply chains, has currently become necessary, and agri-food waste represents a very attractive hub to produce innovative cosmetic extracts. In this paper, an integrated approach considering all the different steps, starting from material selection, extraction, chemical characterization, biological activity evaluation, and environmental impact calculation, was adopted to obtain innovative, sustainable, and effective cosmetic raw materials from food waste. As case report, a supercritical CO2 extract obtained from wild-strawberry-processing waste after jam production (WSWSCO2 extract) was developed. The fatty acids profile of the waste material and WSWSCO2 extract was investigated via a GC–MS method, and mainly polyunsaturated fatty acids (PUFAs) such as linoleic and linolenic acids were detected. Furthermore, the ability of the WSWSCO2 extract to inhibit 5α-reductase type 1 expression in skin fibroblasts was assessed, confirming significant efficacy at the dose of 5 mg/mL. Finally, in view of the eco-sustainability approach, the environmental impact related to WSWSCO2 extract was calculated using a life cycle assessment (LCA) analytical approach, considering different parameters and indicators (e.g., carbon footprint) and verifying the eco-friendly approach in extract development and production. Although further research is needed, for example, to check the full composition of the extract and its effect on skin cells, these results suggest that the WSWSCO2 extract may represent an innovative and sustainable ingredient for cosmetic applications especially in topical preparation for the treatment of some androgenic-related discomfort, such as acne and androgenic alopecia, reflecting the potentiality of the holistic and pioneering approach related to ingredient development presented in this study for the cosmetic sector.

Abstract PR003: Sex-dependent changes in the aged melanoma tumor microenvironment influence metastasis and therapeutic responses

Abstract There is documented sex disparity in cutaneous melanoma incidence and mortality, increasing disproportionately with age and in the male sex. However, the underlying mechanisms remain unclear; while biological sex differences and inherent immune response variability have been assessed in tumor cells, the role of the tumor-surrounding microenvironment, contextually in aging, has been overlooked. Dermal fibroblasts are known to have profound impact on melanoma progression. We examined if both sex-dependent and age-related changes in these fibroblasts can alter the course of melanoma tumor growth, visceral metastasis, and responses to therapy. We find that skin fibroblasts undergo age-mediated sex-differential changes in their proliferation, senescence, ROS production and stress response. We find that aged male fibroblasts drive an invasive, therapy-resistant phenotype in melanoma cells and promote metastasis in aged male mice by increasing AXL expression. Intrinsic aging promotes sex- dimorphic BMP2 secretion exclusively by aged male fibroblasts, which in turn drives the slower-cycling, highly invasive, and therapy-resistant melanoma cell phenotype, characteristic of the aged male TME. Inhibition of BMP2 activity blocks the emergence of invasive phenotype and sensitizes melanoma cells to BRAF/MEK inhibition. Our data provide an integrated view of how host factors such as advancing age and biological sex alter the tumor microenvironment and contribute to disease progression. Bridging this knowledge gap will improve patient stratification and assist in tailoring the therapy and achieve gender-equitable healthcare. Citation Format: Yash Chhabra, Ashani T. Weeraratna. Sex-dependent changes in the aged melanoma tumor microenvironment influence metastasis and therapeutic responses [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr PR003.

Development of an at-line coupling of LC-QTOF-ESI-MS/MS to steroid 5-alpha reductase inhibition assay, a fast bioactive targeting and guided purification of natural complex sample, Impatiens balsamina Linn.

This study provides a rapid and accurate method for screening steroid 5-alpha reductase (S5αR) inhibitors in Impatiens balsamina Linn (IB). using at-line LC-QTOF-ESI-MS/MS coupling S5αR inhibitory assay. IB (Balsaminaceae) is an annual herbaceous plant cultivated in tropical and subtropical regions. It has been used in traditional Chinese and Thai medicine for treatment of hair loss and various skin conditions, potentially through anti-androgenic mechanisms. A combined approach of S5αR inhibitory assay and LC-QTOF-ESI-MS/MS was developed to rapidly screen for target biomarkers and guide their isolation using preparative HPLC. The toxicity of both the extract and isolated biomarkers was evaluated on skin cells, keratinocytes, and fibroblasts. Eight bioactive compounds were identified as two naphthoquinone, two fatty acid derivatives, three nitrogenous compounds and one aromatic derivative. The most potent bioactive markers, identified as 2-methoxy-1,4-naphthoquinone (2MN) and impateinol, were targeted and isolated using preparative HPLC, yielding 5.0 % and 3.5 %, respectively. These compounds exhibited S5αR inhibitory activity higher than that of finasteride drug by 10 and 2 times, respectively. Both the isolated biomarkers and the extract demonstrated a broad therapeutic index. The developed method in this study proved to be both rapid and accurate, making it suitable for screening and targeting S5αR inhibitors in herbal plants or complex matrix samples. It facilitated the fast-guided isolation of bioactive compounds, highlighting its potential for future applications in drug discovery research.

Influence of the Nucleo-Shuttling of the ATM Protein on the Response of Skin Fibroblasts from Marfan Syndrome to Ionizing Radiation.

Marfan syndrome (MFS) is an autosomal dominant connective-tissue disorder affecting multiple systems, such as skeletal, cardiovascular, and ocular systems. MFS is predominantly caused by mutations in the FBN1 gene, which encodes the fibrillin-1 protein, crucial for connective-tissue integrity. FBN1 mutations lead to defective fibrillin, resulting in structurally compromised connective tissues. Additionally, these mutations cause aberrant TGF-β expression, contributing to vascular issues and increased susceptibility to radiation-induced fibrosis. Studies about the potential radiosensitivity of MFS are rare and generally limited to case reports. Here, we aimed to investigate the radiation-induced ATM nucleo-shuttling (RIANS) model to explore the molecular and cellular radiation response in fibroblasts from MFS patients. The results showed that the MFS fibroblast cell lines tested are associated with moderate but significant radiosensitivity, high yield of micronuclei, and impaired recognition of DNA double-strand breaks (DSBs) caused by a diminished RIANS. The diminished RIANS is supported by the sequestration of ATM protein in the cytoplasm not only by mutated FBN1 protein but also by overexpressed TGF-β. This report is the first molecular and cellular characterization of the radiation response of MFS fibroblasts and highlights the importance of the FBN1-TGF-β complex after irradiation.

Aloperine Attenuates UVB-induced Damage in Skin Fibroblasts Via Activating TFE3/Beclin-1-Mediated Autophagy.

Aloperine (ALO) is an important active ingredient in the traditional Chinese medicinal plant Sophora alopecuroides L and has a significant autophagy-stimulating effect. The effect of ALO on cytotoxicity caused by UVB radiation in skin fibroblasts and the potential mechanism remains unclear. The present study aimed to assess the effect of ALO on UVB-induced damage in skin fibroblasts and investigate its possible mechanism. Cell viability, cytotoxicity, caspase-Glo 3/7 activity, apoptosis, and protein expression were measured in UVB-treated skin fibroblasts in the presence or absence of ALO. Autophagy inhibitors (chloroquine and bafilomycin A1) and TFE3 siRNA transfection were used to elucidate the potential mechanisms further. These data demonstrate that ALO attenuated cell viability inhibition, apoptosis, cytotoxicity, and alterations in autophagy-related proteins caused by UVB exposure in skin fibroblasts. ALO stimulates autophagy activation and TFE3 nuclear localization in UVB-treated skin fibroblasts. Furthermore, treatment with autophagy inhibitors and TFE3 siRNA reversed the effects of ALO on UVB-treated skin fibroblasts. These results suggest that ALO protects skin fibroblasts against UVB-induced cytotoxicity by stimulating TFE3/Beclin-1-mediated autophagy.

Investigating p.Ala1035Val in NPC1: New Cellular Models for Niemann-Pick Type C Disease.

Niemann-Pick type C (NPC) is a lysosomal storage disorder (LSD) caused by pathogenic variants in either the NPC1 or NPC2 genes, which encode proteins involved in the lysosomal export of unesterified cholesterol. In patients of Western European descent, the p.Ile1061Thr variant in NPC1 is especially prevalent. However, mounting evidence has positioned p.Ala1035Val as the most common variant in Portugal and the second most prevalent variant worldwide. By analyzing 10 Portuguese NPC patients homozygous for p.Ala1035Val, we found an SNP in cis on position 858 (p.Ile858Val), which we hypothesize could have a disease-modifying effect. To address this query, we created variant-specific in vitro models of NPC by stably transducing NPC1-/- ARPE-19 cells with constructs encoding different fluorescently-tagged variants of NPC1, which we used, alongside patient-derived skin fibroblasts, to investigate lysosomal positioning and the trafficking routes elicited by p.Ile1061Thr and p.Ala1035Val (with and without the p.Ile858Val SNP in cis). Our results corroborate the previously described decrease in p.Ile1061Thr-NPC1 trafficking to the lysosome and suggest a similar, if not worse, scenario for the p.Ala1035Val variant, especially when in cis with p.Ile858Val. This is the first reported functional study addressing the impact of the p.Ala1035Val variant at the cellular level, paving the way for novel therapeutic options.

E'jiao and Cubilose Formula Induced Antioxidant Activity and Improved Collagen Expression of Human Skin Fibroblasts During Oxidation Damage.

As traditional Chinese medicinal materials, E'jiao and cubilose are rich in various bioactive substances with good antioxidant, anti-inflammatory, and immune-regulating effects. To obtain the optimal ratio of synergistic effect between E'jiao and cubilose. The antioxidant capacity of E'jiao and cubilose digestive fluid was evaluated invitro, as well as the intracellular oxidation balance between HSF cells and 3D whole-skin model induced by H2O2. E'jiao, cubilose, and their different ratios of composites had better scavenging ability against free radicals such as DPPH (2,2-Diphenyl-1-picrylhydrazyl). Using HSF cells induced by H2O2 as an oxidative damage model, it was found that the combination of E'jiao and cubilose in a ratio of 2:3 significantly enhanced the activities of SOD, GSH-PX, and CAT enzymes compared to the separate treatments of E'jiao and cubilose, as well as other combination ratios. It effectively reduced the accumulation of reactive oxygen species caused by H2O2 in HSF (Human Skin Fibroblast)cells and protected the integrity of the cells. Further analysis using flow cytometry and a 3D full-thickness skin model revealed that the combination ratio of 2:3 increased the proportion of H2O2-treated cells in the S%+G2% phase from 19.1% + 7.4%-22.1% + 28.8%, helping oxidatively damaged cells partially recover their proliferative capacity. It also promoted the expression of collagen I and collagen IV in the 3D full-thickness skin model, with improvement rates reaching 168.00% and 123.68%, respectively. These findings indicate that the combination of E'jiao and cubilose in a ratio of 2:3 exhibits good synergistic effects, enhancing the ability of cells to resist oxidative damage, promoting cellular renewal and metabolism, and improving skin antiwrinkle capacity.

Enhancement of human native skin fibroblast proliferation in natural salso-bromo-iodic mineral water added to in vitro culture.

The favorable regenerative effects of some mineral waters on wound healing have long been empirically demonstrated. The aim of this experimental study is to investigate the effects of an Italian salso-bromo-iodic mineral water (Rivanazzano, Italy) on an in vitro human native fibroblast culture model to identify any potential regenerative actions. Human native fibroblasts were cultured under different experimental conditions: - Dulbecco's modified Eagle's medium (DMEM) reconstituted with distilled water (control); - DMEM reconstituted with filtered mineral water collected from the spring; - DMEM reconstituted with filtered mineral water collected at the balneotherapy facility; - DMEM reconstituted with filtered, heated mineral water collected at the balneotherapy facility; - DMEM partially replaced with filtered mineral water collected from the spring at different concentrations (10%, 20%, 30%, 40%, 50%); - DMEM partially replaced with filtered, heated mineral water collected at the balneotherapy facility at different concentrations (10%, 20%, 30%, 40%, 50%); - DMEM partially replaced with filtered mineral water collected at the balneotherapy facility at different concentrations (10%, 20%, 30%, 40%, 50%). Cell proliferation and viability were evaluated using spectrophotometric analysis following staining with the XTT Microculture Tetrazolium Assay. Statistical analyses were performed for each experimental condition at 24, 48 and 72 h. The best outcomes were observed in fibroblasts cultured with DMEM partially replaced with filtered mineral water collected from the spring, within the range of 20-50%. Our research results showed that Rivanazzano salso-bromo-iodic mineral water has a stimulating effect on in vitro human native fibroblast cultures. This activity was most pronounced with water collected from the spring, and it decreased with water collected at the balneotherapy facilities. These findings could form the basis for clinical applications in wound healing and balneotherapy.

Presence of mitochondrial dysfunction in a case of Fanconi syndrome with normoglycemic MODY1.

Maturity-onset diabetes of the young 1 (MODY1) is characterized by macrosomia and transient hypoglycemia in neonates, in addition to diabetes mellitus (DM). Only patients with MODY1 harboring a pathogenic variant (c.187C > T; p.R63W) in HNF4A are sure to develop Fanconi syndrome (FS). Here we report the successful diagnosis of MODY1 in a patient harboring p.R63W before confirmation of DM-related hyperglycemia after being alerted to the presence of abnormal mitochondria in a kidney-biopsy specimen revealed by electron microscopy. The patient was born at 39weeks of gestation with macrosomia, elevated levels of liver enzymes, and transient hypoglycemia. At three years of age, proteinuria was found by chance, and further laboratory investigations revealed metabolic acidosis, mild renal dysfunction, hypouricemia, proteinuria, aminoaciduria, and glycosuria. On this basis, we diagnosed the patient as having FS and performed percutaneous renal biopsy. Light microscopy revealed no evidence of proximal tubule disorder, but electron microscopy demonstrated mitochondria with disordered cristae in glomerular podocytes and giant mitochondria in proximal tubules. Mitochondrial nephropathy was suspected, and skin fibroblasts from the patient grown on galactose medium showed decreased oxygen consumption suggestive of mitochondrial dysfunction. Therefore, genetic testing was performed and a pathogenic variant (c.187C > T; p.R63W) in HNF4A was detected. Mitochondrial dysfunction in a Drosophila and murine model of patients with both MODY1 and FS has already been reported, and we detected it in this human MODY1/FS patient on the basis of functional tests and imaging. We believe mitochondrial dysfunction may be involved in the pathogenesis of MODY1/FS.

Potential of epicatechin as antioxidant and antiaging in UV-induced BJ cells by regulating COL1A1, FGF-2, GPX-1, and MMP-1 gene, protein levels, and apoptosis

Background Oxidative stress caused by exposure to ultraviolet (UV) light on the skin can damage deoxyribonucleic acid (DNA) and cause keratinocytes to undergo apoptosis. Endogenous antioxidants which play a role in trapping free radicals are also unable to overcome excess reactive oxygen species (ROS) in the body due to UV exposure, so exogenous antioxidants are needed. Polyphenolic compounds extracted from natural ingredients such as flavonoids, quercetin, and epicatechin have quite strong antioxidant activity. This is influenced by the chemical structure of these compounds which are rich in hydroxyl groups and aromatic groups. This structure allows the compound to become an electron donor so that it can neutralize free radicals. In vitro research was used to see the potential effectiveness of epicatechin as an antiaging and antioxidant. The study aims to confirm the potential of epicatechin as an antiaging by in vitro assay. Methods The viability test of epicatechin on human skin fibroblast (BJ) cells was carried out using the water-soluble tetrazolium (WST) assay. BJ cells were UV-induced as a cell model of premature aging. Epicatechin 6.25, 12.5, and 25 µg/mL were administered to UV-induced BJ cells. The gene expression of Collagen I Alpha 1 (COL1A1), matrix metalloproteinase-1 (MMP-1), fibroblast growth factor-2 (FGF-2), and glutathione peroxidase-1 (GPX-1) were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). Elastin (ELN), hyaluronidase (HAase), cyclooxigenase-2 (COX-2), 8-hydroxydeoxyguanosine (8-OhdG), and melatonin (MT) protein levels were analyzed by enzyme-linked immunosorbent assay (ELISA). The apoptosis of BJ cells was analyzed using flow cytometry. Results Treatment with epicatechin increased relative gene expression including COL1A1 (5.94), FGF-2 (8.34), and GPX-1 (8.09), and also decreased MMP-1 (2.90) relative gene expression compared to the UV-induced BJ cells. Epicatechin also increased levels of ELN (107.7 ng/mg protein) and MT (830 ng/mg protein) levels compared to the UV-induced BJ cells. Epicatechin treatment decreased levels of HAase (505.96 ng/mg protein), COX-2 (33.69 ng/mg protein), and 8-OHdG (97.87 ng/mg protein) compared to the UV-induced BJ cells. Epicatechin also succeeded in maintaining the percentage of live cells and reducing apoptosis, necrotic of UV-induced skin fibroblast cells. Conclusions Epicatechin has the potential to be an antiaging agent by in vitro assay.

Antioxidant Effect of a Plant-Derived Extracellular Vesicles' Mix on Human Skin Fibroblasts: Induction of a Reparative Process.

Plant-Derived Extracellular Vesicles extracellular vesicles (PDEVs) from organic agriculture (without the use of pesticides and microbicides) contain high levels of antioxidants. Organic PDEVs have shown an increased antioxidant power compared to PDEVs from single plants, suggesting a synergistic effect of the bioactives constitutively expressed in the PDEVs from single fruits. With this study, we wanted to investigate the beneficial effects of a mix of PDEVs on human skin cells. We found detectable levels of citric acid, ascorbic acid, glutathione, catalase, and SOD in a mix of PDEVs deriving from five different fruits (grape, red orange, papaya, pomegranate, and tangerine). We then treated H2O2-conditioned fibroblasts with the mix of PDEVs. The results showed that the PDEVs' mixture reverted the H2O2-induced redox imbalance, restoring mitochondrial homeostasis, with a strong reduction of mitochondrial anion superoxide and an increase in sirtuin levels. The antioxidant action was consistent with wound repair on a lesion produced in a fibroblast's monolayer. This result was consistent with an increased level of vimentin and matrix metalloproteinase-9, whose expression is directly related to the efficiency of the reparative processes. These data support a beneficial role of PDEVs in both preventing and treating skin injuries through their potent antioxidant and reparative activities.

Automatic construction of Petri net models for computational simulations of molecular interaction network

Petri nets are commonly applied in modeling biological systems. However, construction of a Petri net model for complex biological systems is often time consuming, and requires expertise in the research area, limiting their application. To address this challenge, we developed GINtoSPN, an R package that automates the conversion of multi-omics molecular interaction network extracted from the Global Integrative Network (GIN) into Petri nets in GraphML format. These GraphML files can be directly used for Signaling Petri Net (SPN) simulation. To demonstrate the utility of this tool, we built a Petri net model for neurofibromatosis type I. Simulation of NF1 gene knockout, compared to normal skin fibroblast cells, revealed persistent accumulation of Ras-GTPs as expected. Additionally, we identified several other genes substantially affected by the loss of NF1’s function, exhibiting individual-specific variability. These results highlight the effectiveness of GINtoSPN in streamlining the modeling and simulation of complex biological systems.