Abstract Background Taraxasterol, a pentacyclic triterpenoid compound, has been widely used in traditional and modern medicine because of its pharmacological properties such as anti-inflammatory, antioxidative and antitumor effects. 2,3-oxidosqualene cyclase (OSC) is highly important for the generation of phytosterols and triterpenoid compounds and the structural diversity of natural products. However, the specific role of TmOSCs in the taraxasterol biosynthesis pathway has not yet been precisely resolved. Results In this study, 10 TmOSC gene family members were identified via the Taraxacum mongolicum (dandelion) genome and classified into three subgroups. Phylogenetic and collinearity analyses revealed evolutionary conservation among OSC proteins from Asteraceae species. RNA-seq data analysis revealed that TmOSC8 and TmOSC10 were highly expressed in nutrient-containing tissues. In addition, methyl jasmonate (MeJA) and abscisic acid (ABA) significantly induced the expression of TmOSC3, and the change in its relative expression was consistent with the taraxasterol content. The relative expression level of the TmOSC8-overexpressioning line was significantly increased by approximately 20 fold compared with that of the wild type, and the content of taraxasterol was significantly increased to 3 fold greater than that of the wild type. Conclusion This study systematically analyzed the evolutionary characteristics and expression patterns of the TmOSC gene family, suggested potential key roles of TmOSC3 and TmOSC8 in sterol synthesis and stress response, and provided a preliminary theoretical basis for the metabolic engineering of medicinal components in dandelion.
Gelatin methacryloyl (GelMA), a biomaterial widely used in tissue engineering, exhibits excellent biocompatibility and cell adhesion properties. However, its poor mechanical strength and functional monotony restrict broader clinical applications of this material. In this study, we introduced sodium acrylate (SA) and tannic acid (TA) into the GelMA system via a two-step crosslinking strategy, successfully fabricating a GelMA/SA–TA (GST) composite hydrogel that achieved dual enhancement of mechanical and antioxidant properties. The incorporation of SA and TA significantly improved the mechanical performance of the hydrogel, which exhibited a maximum tensile modulus of 31.83 ± 2.84 kPa. At the same time, TA endowed the hydrogel with exceptional antioxidant ability, resulting in a free radical scavenging rate of 89.93% ± 0.9% in vitro. Biological tests revealed that the GST hydrogel effectively alleviated oxidative stress damage in rat Schwann cells (RSC96) by suppressing the generation of reactive oxygen species (ROS) and promoting the secretion of brain-derived neurotrophic factor (BDNF). This work presents the first report of an antioxidant hydrogel capable of protecting Schwann cells without compromising their mechanical integrity, highlighting its transformative potential for peripheral nerve injury repair. The synergistic SA–TA modification strategy provides new insights into the design of multifunctional biomaterials for neural regeneration applications.
Daniel Sanchez‐Morillo, Antonio León‐Jiménez, María Guerrero‐Chanivet
et al.
Abstract Engineered stone silicosis (ESS), primarily caused by inhaling respirable crystalline silica, poses a significant occupational health risk globally. ESS has no effective treatment and presents a rapid progression from simple silicosis (SS) to progressive massive fibrosis (PMF), with respiratory failure and death. Despite the use of diagnostic methods like chest x‐rays and high‐resolution computed tomography, early detection of silicosis remains challenging. Since routine blood tests have shown promise in detecting inflammatory markers associated with the disease, this study aims to assess whether routine blood biomarkers, coupled with machine learning techniques, can effectively differentiate between healthy individuals, subjects with SS, and PMF. To this end, 107 men diagnosed with silicosis, ex‐workers in the engineered stone (ES) sector, and 22 healthy male volunteers as controls not exposed to ES dust were recruited. Twenty‐one primary biochemical markers derived from peripheral blood extraction were obtained retrospectively from clinical hospital records. Relief‐F features selection technique was applied, and the resulting subset of 11 biomarkers was used to build five machine learning models, demonstrating high performance with sensitivities and specificities in the best case greater than 82% and 89%, respectively. The percentage of lymphocytes, the angiotensin‐converting enzyme, and lactate dehydrogenase indexes were revealed, among others, as blood biomarkers with significant cumulative importance for the machine learning models. Our study reveals that these biomarkers could detect a chronic inflammatory status and potentially serve as a supportive tool for the diagnosis, monitoring, and early detection of the progression of silicosis.
Anil Philip Kunnath, Mohamed Suodha Suoodh, Dinesh Kumar Chellappan
et al.
The global issue of antimicrobial resistance poses significant challenges to public health. The World Health Organization (WHO) has highlighted it as a major global health threat, causing an estimated 700,000 deaths worldwide. Understanding the multifaceted nature of antibiotic resistance is crucial for developing effective strategies. Several physiological and biochemical mechanisms are involved in the development of antibiotic resistance. Bacterial cells may escape the bactericidal actions of the drugs by entering a physiologically dormant state known as bacterial persistence. Recent findings in this field suggest that bacterial persistence can be one of the main sources of chronic infections. The antibiotic tolerance developed by the persister cells could tolerate high levels of antibiotics and may give rise to persister offspring. These persister offspring could be attributed to antibiotic resistance mechanisms, especially in chronic infections. This review attempts to shed light on persister-induced antibiotic resistance and the current therapeutic strategies.
The <i>Brassica oleracea</i> L. species embrace important horticultural crops, such as broccoli, cauliflower, and cabbage, which are highly valued for their beneficial nutritional effects. However, the complexity of flower emasculation in these species has forced breeders to adopt biotechnological approaches such as somatic hybridization to ease hybrid seed production. Protoplasts entail a versatile tool in plant biotechnology, supporting breeding strategies that involve genome editing and hybridization. This review discusses the use of somatic hybridization in <i>B. oleracea</i> L. as a biotechnological method for developing fusion products with desirable agronomic traits, particularly cytoplasmic male sterile (CMS) condition. These CMS lines are critical for implementing a cost-effective, efficient, and reliable system for producing F1 hybrids. We present recent studies on CMS systems in <i>B. oleracea</i> L. crops, providing an overview of established models that explain the mechanisms of CMS and fertility restoration. Additionally, we emphasize key insights gained from protoplast fusion applied to <i>B. oleracea</i> L. breeding. Key steps including pre-treatments of donor plants, the main tissues used as sources of parental protoplasts, methods for obtaining somatic hybrids and cybrids, and the importance of establishing a reliable plant regeneration method are discussed. Finally, the review explores the incorporation of genome editing technologies, such as CRISPR-Cas9, to introduce multiple agronomic traits in <i>Brassica</i> species. This combination of advanced biotechnological tools holds significant promise for enhancing <i>B. oleracea</i> breeding programs in the actual climate change context.
Bone regeneration and repair are complex processes with the potential of added complications, like delayed repair, fracture non-union, and post-surgical infections. These conditions remain a challenge globally, pressurizing the economy and patients suffering from these conditions. Applications of nanotechnology (NBT) in the field of medicine have provided a medium for several approaches to support these global challenges. Tissue engineering is one such field that has been on the rise in the last three decades through the utilization of NBT for addressing the challenges related to bone regeneration. First, NBT enables the formation of scaffolds at the nanoscale needed for bone tissue engineering (BTE) using natural and synthetic polymers, as well as with minerals and metals. Then, it aids the development of the nano-formulation strategized to deliver antimicrobial drugs and/or growth factors through various ways to enhance bone repair through the scaffold. Third, NBT facilitates the use of specialized nanoparticles to image and track cellular events in vitro as well as in vivo. This review is an effort to bring together the current knowledge in the field of BTE and present the scope of ever-evolving NBT, a contribution towards precision medicine.
Abdul Sattar, Ahmad Sher, Mohammed A. S. Abourehab
et al.
Arsenic (As) contamination in the soil adversely affects crop productivity, grain quality, and human health. A pot experiment was performed to assess the sole and combined effects of Si and biochar on growth, physiological and antioxidant defense mechanisms, yield, and grain quality of maize. Soil treatments comprised of control (no treatment), As, Si, biochar, Si + biochar, As + Si, As + biochar, and As + Si + biochar placed in a completely randomized design with three replications. As toxicity significantly reduced chlorophyll a (5.18%), chlorophyll b (33.87%), chlorophyll a + b (11.67%), and primary metabolites [soluble protein (54.93%), amino acids (24.85%), total soluble sugars (39.77%), and phenolic contents (25.88%)], while increasing the activities of enzymatic antioxidants such as superoxide dismutase (SOD) by 43.51%, peroxidase (POD) by 47.93%, catalase (CAT) by 47.98%, and ascorbate peroxidase (APX) by 59.02%, as well as that of lipid peroxidation in the leaves of maize. In addition, As contamination reduced the grain yield and yield-related attributes relative to the respective controls. Among the soil applications, the interactive effect of Si and biochar improved maize grain yield (12.12%) by triggering activities of enzymatic antioxidants and proline contents and reducing the H2O2 and MDA contents. The combined application of Si and biochar enhanced the Si contents in shoots of the control and As-contaminated plants, while significantly reducing As concentration in shoots (69%) and grains (142%). In conclusion, the combined application of Si and biochar was found to be a fruitful soil amendment strategy to improve the yield of maize and reduce the toxic limit of As under As-contaminated soil. The results of this study may be useful for the cultivation of food crops under AS-contaminated soils, but before commercial recommendation, more trails are required under field conditions.
Tomasz Ligor, Przemysław Adamczyk, Tomasz Kowalkowski
et al.
Bladder cancer is one of most common types of cancer diagnosed in the genitourinary tract. Typical tests are costly and characterized by low sensitivity, which contributes to a growing interest in volatile biomarkers. Head space solid phase microextraction (SPME) was applied for the extraction of volatile organic compounds from urine samples, and gas chromatography time of flight mass spectrometry (GC×GC TOF MS) was used for the separation and detection of urinary volatiles. A cohort of 40 adult patients with bladder cancer and 57 healthy persons was recruited. Different VOC profiles were obtained for urine samples taken from each group. Twelvecompounds were found only in the samples from theBC group.The proposed candidate biomarkers are butyrolactone; 2-methoxyphenol; 3-methoxy-5-methylphenol; 1-(2,6,6-trimethylcyclohexa-1,3-dien-1-yl)-2-buten-1-one; nootkatone and 1-(2,6,6-trimethyl-1-cyclohexenyl)-2-buten-1-one.Since most of the studies published in the field are proving the potential of VOCs detected in urine samples for the screening and discrimination of patients with bladder cancer from healthy, but rarely presenting the identity of proposed biomarkers, our study represents a novel approach.
Objective: Combined hepatocellular-cholangiocarcinoma (cHCC-CC) is a rare type of primary liver cancer with
characteristics of both hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC). The pathogenesis of cHCCCC
is poorly understood due to a shortage of suitable in vitro models. Due to scarce availability of human liver tissue,
induced pluripotent stem cells (iPSCs) are a useful alternative source to produce renewable liver cells. For use in the
development of liver pathology models, here we successfully developed and evaluated iPSCs from liver fibroblasts of
a patient with cHCC-CC.
Materials and Methods: In this experimental study, human liver fibroblasts (HLFs) were obtained from the liver biopsy of
a 69-year-old male patient with cHCC-CC and transduced with a retroviral cocktail that included four factors - OCT4, SOX2,
KLF4, and c-MYC (OSKM). Pluripotency of the iPSCs was determined by alkaline phosphatase (AP) staining, quantitative
real-time polymerase chain reaction (PCR), and immunofluorescence. We induced in vitro embryoid body (EB) formation
and performed an in vivo teratoma assay to confirm their differentiation capacity into the three germ layers.
Results: HLF iPSCs derived from the cHCC-CC patient displayed typical iPSC-like morphology and pluripotency
marker expression. The proficiency of the iPSCs to differentiate into three germ layers was assessed both in vitro and
in vivo. Compared to normal control iPSCs, differentiated HLF iPSCs showed increased expressions of HCC markers
alpha-fetoprotein (AFP) and Dickkopf-1 (DKK1) and the CC marker cytokeratin 7 (CK7), and a decreased expression
of the CC tumour suppressor SRY-related HMG-box 17 (SOX17).
Conclusion: We established HLF iPSCs using liver fibroblasts from a patient with cHCC-CC for the first time. The HLF
iPSCs maintained marker expression in the patient when differentiated into EBs. Therefore, HLF iPSCs may be a sustainable
cell source for modelling cHCC-CC and beneficial for understanding liver cancer pathology and developing therapies for
cHCC-CC treatment.
Noureddine Bencheikh, Amine Elbouzidi, Loubna Kharchoufa
et al.
Kidney disease is one of the most common health problems and kidney failure can be fatal. It is one of the health disorders associated with extreme pain and discomfort in patients. In developing countries, such as Morocco where socioeconomic and sanitary conditions are precarious, medicinal plants are considered the primary source of medication. In the present work an ethnobotanical survey was conducted in a remote area of North-Eastern Morocco and we focused on (1) establishing a record of medicinal plants used traditionally by local people to treat kidney diseases and (2) correlate the obtained ethnomedical use with well-studied pharmacological evidence. From February 2018 to January2020, information was gathered from 488 informants using semi-structured questionnaires. The data were analyzed using three quantitative indices: The use value (UV), family use value (FUV), and informant consensus factor (ICF). A total of 121 plant species belonging to 57 botanical families were identified to treat kidney diseases. The families most represented were Asteraceae (14 species), followed by Lamiaceae (12 species) and Apiaceae (10 species). The most commonly used plant parts were leaves, followed by the whole plant and they were most commonly prepared by decoction and infusion. The highest value of the (UV) index was attributed to <i>Herniaria hirsuta</i> L. (UV = 0.16), and the highest family use value (FUV) was assigned to Caryophyllaceae with (FUV = 0.163). Regarding the informant consensus factor (ICF), this index’s highest values were recorded for kidney stones (ICF = 0.72). The use of 45% of the selected plants were validated based on literature review. This study helped document and preserve crucial traditional plant knowledge of 121 plant species used to treat kidney problems that can be used in the search for new biologically active compounds through more upcoming pharmacological studies.
Maliheh Vahidinasab, Lars Lilge, Aline Reinfurt
et al.
Abstract Background Plipastatin is a potent Bacillus antimicrobial lipopeptide with the prospect to replace conventional antifungal chemicals for controlling plant pathogens. However, the application of this lipopeptide has so far been investigated in a few cases, principally because of the yield in low concentration and unknown regulation of biosynthesis pathways. B. subtilis synthesizes plipastatin by a non-ribosomal peptide synthetase encoded by the ppsABCDE operon. In this study, B. subtilis 3NA (a non-sporulation strain) was engineered to gain more insights about plipastatin mono-production. Results The 4-phosphopantetheinyl transferase Sfp posttranslationally converts non-ribosomal peptide synthetases from inactive apoforms into their active holoforms. In case of 3NA strain, sfp gene is inactive. Accordingly, the first step was an integration of a repaired sfp version in 3NA to construct strain BMV9. Subsequently, plipastatin production was doubled after integration of a fully expressed degQ version from B. subtilis DSM10T strain (strain BMV10), ensuring stimulation of DegU-P regulatory pathway that positively controls the ppsABSDE operon. Moreover, markerless substitution of the comparably weak native plipastatin promoter (P pps ) against the strong constitutive promoter P veg led to approximately fivefold enhancement of plipastatin production in BMV11 compared to BMV9. Intriguingly, combination of both repaired degQ expression and promoter exchange (P pps ::P veg ) did not increase the plipastatin yield. Afterwards, deletion of surfactin (srfAA-AD) operon by the retaining the regulatory comS which is located within srfAB and is involved in natural competence development, resulted in the loss of plipastatin production in BMV9 and significantly decreased the plipastatin production of BMV11. We also observed that supplementation of ornithine as a precursor for plipastatin formation caused higher production of plipastatin in mono-producer strains, albeit with a modified pattern of plipastatin composition. Conclusions This study provides evidence that degQ stimulates the native plipastatin production. Moreover, a full plipastatin production requires surfactin synthetase or some of its components. Furthermore, as another conclusion of this study, results point towards ornithine provision being an indispensable constituent for a plipastatin mono-producer B. subtilis strain. Therefore, targeting the ornithine metabolic flux might be a promising strategy to further investigate and enhance plipastatin production by B. subtilis plipastatin mono-producer strains.
Behshad Eskandari, Maliheh Safavi, Seyede Nargess Sadati Lamardi
et al.
Nowadays, cancer is the second prevalent cause of mortality after cardiovascular diseases in developed and the third one in developing countries. Adverse effects of chemotherapeutic agents bring the necessity of investigating about new medications with fewer side effects. Daphne L. genus is one of the natural sources with valuable reported anticancer effects. This study aimed to assess the cytotoxic effect of some extracts from the aerial parts of Daphne pontica collected from North of Iran on cancer cell lines. Extraction of the plant material was performed by maceration (3×72 h) of 200 g of sample with petroleum ether, chloroform, ethyl acetate, and methanol, respectively. The total extract was also obtained by maceration of the sample with 80% ethanol. Different concentrations of the dried extracts were prepared to assess their cytotoxic effect by 24 h incubation of cell lines with different extracts and then MTT (dimethyl thiazolyl diphenyl tetrazolium) assay on three cancerous cell lines (MDA-MB-231, MCF-7 and T47D), performed in triplicate. IC50 was then estimated from curves constructed by plotting cell survival (%) versus sample concentration (µg/ml). Results indicated that ethyl acetate fraction of D. pontica had the most potent cytotoxic effect in MTT assay with IC50=977.46 µg/ml; while other fractions were weaker in toxicity (IC50>1000 µg/ml). By comparing to potent cytotoxic effects of other Daphne species, it seems that the cytotoxic properties of D. pontica is different from other species of this genus since according to this study, no significant antineoplastic properties against the three breast cancer cell lines were determined. Further studies on other pharmacological activities of this plant are recommended.
Summary: Proper transcriptome reprogramming is critical for hosts to launch an effective defense response upon pathogen attack. How immune-related genes are regulated at the posttranscriptional level remains elusive. We demonstrate here that P-bodies, the non-membranous cytoplasmic ribonucleoprotein foci related to 5′-to-3′ mRNA decay, are dynamically modulated in plant immunity triggered by microbe-associated molecular patterns (MAMPs). The DCP1-DCP2 mRNA decapping complex, a hallmark of P-bodies, positively regulates plant MAMP-triggered responses and immunity against pathogenic bacteria. MAMP-activated MAP kinases directly phosphorylate DCP1 at the serine237 residue, which further stimulates its interaction with XRN4, an exonuclease executing 5′-to-3′ degradation of decapped mRNA. Consequently, MAMP treatment potentiates DCP1-dependent mRNA decay on a specific group of MAMP-downregulated genes. Thus, the conserved 5′-to-3′ mRNA decay elicited by the MAMP-activated MAP kinase cascade is an integral part of plant immunity. This mechanism ensures a rapid posttranscriptional downregulation of certain immune-related genes that may otherwise negatively impact immunity. : Yu et al. show that P-body core components DCP1 and DCP2 positively regulate plant pattern-triggered immunity. DCP1 phosphorylation by immune-activated MAPKs contributes to P-body disassembly and mRNA decay on a subset of immune-regulated genes, revealing mRNA-decay-mediated posttranscriptional regulation is an integral part of plant immunity. Keywords: plant immunity, microbe-associated molecular patterns, processing body, mRNA decay, mRNA decapping complex, MAP kinases