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Frontiers in Bioscience (Landmark... Jun 2024L-Theanine, a nonproteinogenic amino acid derived from green tea, is being recognized as an anti-cancer candidate. However, it's roles in the development of cancer...
BACKGROUND
L-Theanine, a nonproteinogenic amino acid derived from green tea, is being recognized as an anti-cancer candidate. However, it's roles in the development of cancer chemoresistance is still unknown and the molecular mechanism is urgently to be explored.
METHODS
The effects of L-Theanine on lung cancer chemoresistance were validated by Cell Counting Kit-8 (CCK-8) assay, transwell assay, and tumor spheroid formation assay; the expression of proteins was detected by using polymerase chain reaction (PCR) and western blotting. RNA-sequencing (RNA-seq) and bioinformatics analysis were used to identify differentially expressed genes induced by L-Theanine. knockdown and overexpression were constructed by using a lentivirus-mediated transfection system.
RESULTS
L-Theanine improved the chemoresistance to -diamminedichloroplatinum (DDP) and inhibited stemness of DDP-resistant lung cancer cells but not non-resistant lung cancer cells. The results from RNA-seq analysis showed that STAT3/NOTCH1 pathway was a potential dominant signaling involved in L-Theanine improving the chemoresistance in DDP-resistant lung cancer. Mechanistically, L-Theanine impeded migration and stemness activation of DDP-resistant lung cancer cells via regulating the expression of STAT3/NOTCH1/BMAL1 signaling-induced stemness markers as well as inhibiting the expression levels of drug resistance-related genes. In addition, a combination of L-Theanine and Stat3 blockade synergistically improved the chemoresistance in DDP-resistant lung cancer.
CONCLUSION
L-Theanine improves the chemoresistance by regulating STAT3/NOTCH1/BMAL1 signaling, reducing stemness, and inhibiting the migration of DDP-resistant lung cancer cells. The finding might provide some evidence for therapeutic options in overcoming the chemoresistance in cancers, including lung cancer.
Topics: Humans; Glutamates; Drug Resistance, Neoplasm; Lung Neoplasms; Cisplatin; STAT3 Transcription Factor; Signal Transduction; Receptor, Notch1; Cell Line, Tumor; ARNTL Transcription Factors; Antineoplastic Agents; Gene Expression Regulation, Neoplastic; A549 Cells; Cell Movement
PubMed: 38940036
DOI: 10.31083/j.fbl2906226 -
Frontiers in Bioscience (Landmark... Jun 2024Existing animal models for testing therapeutics in the skin are limited. Mouse and rat models lack similarity to human skin in structure and wound healing mechanism....
BACKGROUND
Existing animal models for testing therapeutics in the skin are limited. Mouse and rat models lack similarity to human skin in structure and wound healing mechanism. Pigs are regarded as the best model with regards to similarity to human skin; however, these studies are expensive, time-consuming, and only small numbers of biologic replicates can be obtained. In addition, local-regional effects of treating wounds that are closely adjacent to one-another with different treatments make assessment of treatment effectiveness difficult in pig models. Therefore, here, a novel nude mouse model of xenografted porcine hypertrophic scar (HTS) cells was developed. This model system was developed to test if supplying hypo-pigmented cells with exogenous alpha melanocyte stimulating hormone (α-MSH) will reverse pigment loss .
METHODS
Dyschromic HTSs were created in red Duroc pigs. Epidermal scar cells (keratinocytes and melanocytes) were derived from regions of hyper-, hypo-, or normally pigmented scar or skin and were cryopreserved. Dermal fibroblasts (DFs) were isolated separately. Excisional wounds were created on nude mice and a grafting dome was placed. DFs were seeded on day 0 and formed a dermis. On day 3, epidermal cells were seeded onto the dermis. The grafting dome was removed on day 7 and hypo-pigmented xenografts were treated with synthetic α-MSH delivered with microneedling. On day 10, the xenografts were excised and saved. Sections were stained using hematoxylin and eosin hematoxylin and eosin (H&E) to assess xenograft structure. RNA was isolated and quantitative real-time polymerase chain reaction (qRT-PCR) was performed for melanogenesis-related genes , , and .
RESULTS
The seeding of HTSDFs formed a dermis that is similar in structure and cellularity to HTS dermis from the porcine model. When hyper-, hypo-, and normally-pigmented epidermal cells were seeded, a fully stratified epithelium was formed by day 14. H&E staining and measurement of the epidermis showed the average thickness to be 0.11 ± 0.07 µm 0.06 ± 0.03 µm in normal pig skin. Hypo-pigmented xenografts that were treated with synthetic α-MSH showed increases in pigmentation and had increased gene expression of , , and compared to untreated controls (TYR: 2.7 ± 1.1 0.3 ± 1.1; TYRP1: 2.6 ± 0.6 0.3 ± 0.7; DCT 0.7 ± 0.9 0.3 ± 1-fold change from control; n = 3).
CONCLUSIONS
The developed nude mouse skin xenograft model can be used to study treatments for the skin. The cells that can be xenografted can be derived from patient samples or from pig samples and form a robust dual-skin layer containing epidermis and dermis that is responsive to treatment. Specifically, we found that hypo-pigmented regions of scar can be stimulated to make melanin by synthetic α-MSH .
Topics: Animals; Mice, Nude; Cicatrix, Hypertrophic; Mice; Disease Models, Animal; Swine; alpha-MSH; Humans; Skin; Fibroblasts; Melanocytes; Keratinocytes; Transplantation, Heterologous; Wound Healing; Skin Pigmentation
PubMed: 38940034
DOI: 10.31083/j.fbl2906230 -
Frontiers in Bioscience (Landmark... May 2024Osteosarcoma (OS) is a primary malignant bone tumor in the pediatric and adolescent populations. Long non-coding RNAs (LncRNAs), such as plasma-cytoma variant...
BACKGROUND
Osteosarcoma (OS) is a primary malignant bone tumor in the pediatric and adolescent populations. Long non-coding RNAs (LncRNAs), such as plasma-cytoma variant translocation 1 (PVT1), have emerged as significant regulators of OS metastasis. Recent studies have indicated that activation of signal transducer and activator of transcription 3 (STAT3) signaling, which might be controlled by PVT1, inhibits ferroptosis to promote the malignant progression of cancer. Therefore, the present study aimed to determine the role of PVT1 in OS pathogenesis and investigate whether PVT1 affects OS progression by regulating STAT3/GPX4 pathway-mediated ferroptosis.
METHODS
The human OS cell line MG63 were transfected with sh-PVT1 plasmid to inhibit PVT1 expression, with or without co-transfection with a STAT3 overexpression plasmid. The expression of PVT1 was determined by real-time quantitative polymerase chain reaction (RT-qPCR). The proliferation, migration, invasion, and apoptosis of MG63 cells were determined using the cell counting kit-8 (CCK8), Transwell assay, and flow cytometry. The levels of malondialdehyde (MDA), Fe2+, and glutathione (GSH) were determined by ELISA kits, whereas reactive oxygen species (ROS) level was determined by immunofluorescence. The protein expression levels of STAT3, p-STAT3, and glutathione peroxidase 4 (GPX4) were detected by western blot (WB).
RESULTS
PVT1 expression was significantly increased in MG63 cells. When knocking down PVT1 with sh-PVT1 plasmid, the proliferation, migration, and invasion of MG63 cells were markedly inhibited, while the rate of apoptosis was upregulated. Further investigation revealed that MG63 cells with PVT1 knockdown exhibited elevated levels of MDA, Fe2+, and ROS. In addition, the inhibition of PVT1 expression resulted in decreased levels of GSH and inhibited expression of p-STAT3 and GPX4. When sh-PVT1 was co-transfected with STAT3 overexpression plasmid in MG63 cells, the increased levels of MDA, Fe2+, and ROS were downregulated, and the decreased expressions of GSH, p-STAT3, and GPX4 were upregulated.
CONCLUSION
PVT1 promotes OS metastasis by activating the STAT3/GPX4 pathway to inhibit ferroptosis. Targeting PVT1 might be a novel therapeutic strategy for OS treatment.
Topics: Humans; Osteosarcoma; RNA, Long Noncoding; Ferroptosis; STAT3 Transcription Factor; Cell Line, Tumor; Bone Neoplasms; Phospholipid Hydroperoxide Glutathione Peroxidase; Cell Proliferation; Reactive Oxygen Species; Signal Transduction; Cell Movement; Disease Progression; Apoptosis; Gene Expression Regulation, Neoplastic
PubMed: 38940027
DOI: 10.31083/j.fbl2906207 -
Frontiers in Bioscience (Landmark... Jun 2024Nonalcoholic fatty liver disease (NAFLD) is a prevalent condition characterized by hepatic fat accumulation, often progressing to severe liver injury, for which approved...
BACKGROUND
Nonalcoholic fatty liver disease (NAFLD) is a prevalent condition characterized by hepatic fat accumulation, often progressing to severe liver injury, for which approved treatments are currently lacking. This study explores the potential therapeutic impact of alpha-lipoic acid (ALA), a natural compound crucial in lipid metabolism, on NAFLD using an model.
METHODS
HepG2 cells were treated with a palmitic acid:oleic acid (PA:OA) mixture, representing a cellular model of steatosis. Subsequent treatment with ALA at concentrations of 1 µM and 5 µM aimed to evaluate its effects on lipid content and metabolism. Real-time polymerase chain reaction (PCR), BODIPY staining, cytofluorimetric analysis, and lipidomics were used to assess gene expression, lipid droplet accumulation, and fatty acid profiles.
RESULTS
Our results showed that ALA significantly reduced lipid droplets in PA:OA-treated HepG2 cells, with a concentration-dependent effect. Analysis of fatty acid profiles demonstrated a decrease in palmitic acid levels with ALA treatment, while oleic acid reduction was observed only at the higher concentration. Moreover, ALA modulated the expression of genes involved in cholesterol biosynthesis and low-density lipoprotein (LDL) metabolism, indicating a potential role in lipid homeostasis. Further insights into molecular mechanisms revealed that ALA modulated peroxisome proliferator activated receptors (PPARs), specifically PPAR-alpha and PPAR-gamma, involved in fatty acid metabolism and insulin sensitivity. Finally, ALA counteracted the overexpression of thermogenic genes induced by exogenous fatty acids, suggesting a regulatory role in energy dissipation pathways.
CONCLUSION
In conclusion, this study highlights ALA as a therapeutic agent in mitigating lipid accumulation and dysregulation in NAFLD.
Topics: Humans; Thioctic Acid; Hep G2 Cells; Lipid Metabolism; Non-alcoholic Fatty Liver Disease; Oleic Acid; Palmitic Acid; Gene Expression Regulation; Fatty Acids; PPAR gamma; Lipid Droplets; PPAR alpha; Uncoupling Protein 2
PubMed: 38940024
DOI: 10.31083/j.fbl2906209 -
Frontiers in Bioscience (Landmark... Jun 2024The objective of this research was to identify differentially expressed genes (DEGs) related to ferroptosis in the annulus fibrosus (AF) during intervertebral disc...
BACKGROUND
The objective of this research was to identify differentially expressed genes (DEGs) related to ferroptosis in the annulus fibrosus (AF) during intervertebral disc degeneration (IDD).
METHODS
We analyzed gene data from degenerated and normal AF obtained from the GSE70362 and GSE147383 datasets. An analysis to determine the functional significance of the DEGs was conducted, followed by the creation of a network illustrating the interactions between proteins. We further analyzed the immune infiltration of the DEGs and determined the hub DEGs using LASSO regression analysis. Finally, we identified the hub ferroptosis-related DEGs (FRDEGs) and verified their expression levels using Real-time quantitative polymerase chain reaction (RT-qPCR), Western blot, Immunohistochemical Staining (IHC), and Immunofluorescence (IF).
RESULTS
By analyzing the GSE70362 and GSE147383 datasets, we identified 118 DEGs. In degenerative AF groups, we observed a significant increase in immune infiltration of resting memory CD4+ T cells. LASSO regression analysis revealed 9 hub DEGs. The construction of a Receiver Operating Characteristic (ROC) curve yielded an Area Under the Curve (AUC) value of 0.762. Furthermore, we found that is a hub gene related to ferroptosis. Our examination of immune infiltration indicated that primarily influences macrophage M0 in different immune cell expression groups. Finally, our observations revealed a marked upregulation of expression in the degenerated annulus fibrosus tissue.
CONCLUSION
Our findings indicate an upsurge in levels within degenerative AF, potentially playing a crucial role in the exacerbation of IDD. These findings provide a foundation for further exploration of the pathological mechanisms underlying IDD and offer potential drug targets for intervention.
Topics: Humans; Annulus Fibrosus; Computational Biology; Databases, Genetic; Ferroptosis; Gene Expression Profiling; Gene Regulatory Networks; Intervertebral Disc; Intervertebral Disc Degeneration; Protein Interaction Maps; Glutathione Transferase
PubMed: 38940022
DOI: 10.31083/j.fbl2906224 -
Frontiers in Bioscience (Scholar... Jun 2024Sickle cell disease (SCD) is a major heritable genetic disease in sub-Saharan Africa, including Mauritania. Fetal hemoglobin (HbF) can affect the pathophysiology,...
BACKGROUND
Sickle cell disease (SCD) is a major heritable genetic disease in sub-Saharan Africa, including Mauritania. Fetal hemoglobin (HbF) can affect the pathophysiology, moderate the clinical course, and offer prospects for curative treatment of SCD. This study aimed to investigate the influence of single nucleotide polymorphisms (SNPs) in the gene on the levels of HbF and hematological parameters in Mauritanian sickle cell () patients.
METHODS
Complete blood count was assessed in 565 patients suspected to have SCD. Polymerase chain reaction (PCR)-restriction fragment length polymorphism was performed to identify the , and sequencing was used for genotyping three SNPs: () and () in the intron 2 and () in the regions of the gene in 50 sickle cell patients.
RESULTS
The prevalence of HbSS among the study population was 8.8% (50/565), and the mean (± standard deviation) of HbF level was 15.0% (± 6.0%). Sequencing showed the presence of three genotypes: AA (13.6%), AG (46.6%), GG (39.6%) in rs4671393; CC (17.6%), CT (48.7%), and TT (33.6%) in . All samples from HbSS individuals displayed a wild-type genotype in the rs1052520 allele. The prevalence of minor alleles () and () were 37% and 39%, respectively. There was a statistically significant association ( = 0.034) between rs4671393 SNP and elevated HbF (mean 12.72 ± 6.26%).
CONCLUSIONS
The study of three SNPs in the locus in Mauritanian patients with SCD showed a significant association of allele with the HbF level. Further research is needed to explore additional SNPs in the locus and investigate other genetic markers reported to modulate HbF levels, such as and , to improve the management of this potentially life-threatening condition in Mauritania.
Topics: Humans; Fetal Hemoglobin; Anemia, Sickle Cell; Polymorphism, Single Nucleotide; Female; Male; Adult; Repressor Proteins; Mauritania; Genotype; Nuclear Proteins; Adolescent; Carrier Proteins; Young Adult; Child
PubMed: 38939975
DOI: 10.31083/j.fbs1602011 -
Journal of Parasitology Research 2024Human toxocariasis (HT) is a zoonotic disease with a global expansion. Contaminated soil with spp. eggs is the main source of human infection, which may lead to severe...
BACKGROUND
Human toxocariasis (HT) is a zoonotic disease with a global expansion. Contaminated soil with spp. eggs is the main source of human infection, which may lead to severe complications depending on the organs invaded by migrating larvae.
AIM
This study is aimed at eliciting the prevalence of spp. eggs in public parks in Zahedan, southeast Iran, and providing new insight into the soil contamination rate in this area using microscopic and molecular methods.
METHODS
Based on five municipal districts, 240 soil samples were collected from public parks and playgrounds in Zahedan. The modified Sheather's flotation technique was employed to isolate spp. eggs from the soil, followed by microscopic assessment and molecular evaluation of internal transcribed spacer 1 and 2 ribosomal deoxyribonucleic acid (ITS1 and 2 rDNA) using nested polymerase chain reaction (nested PCR) to identify the presence of spp. eggs. The Sanger sequence was used to differentiate the species. Subsequently, all the sequenced data were blasted and compared with other sequences available in the GenBank.
RESULTS
Out of 240 soil samples collected, 7 (2.9%) samples were identified to contain spp. eggs using Sheather's flotation and microscopic techniques. Meanwhile, 19 (7.9%) samples were positive using nested PCR. According to the Sanger sequencing analysis findings, all positive samples were contaminated with .
CONCLUSION
As evidenced by the obtained results, only species were detected in public parks and playgrounds in Zahedan; therefore, control and prevention programs against this species should be considered in human and animal communities.
PubMed: 38939904
DOI: 10.1155/2024/2132696 -
JACC. Advances Aug 2023Prior studies of COVID-19 cardiovascular sequelae include diagnoses made within 4 weeks, but the World Health Organization definition for "postacute phase" is >3 months.
BACKGROUND
Prior studies of COVID-19 cardiovascular sequelae include diagnoses made within 4 weeks, but the World Health Organization definition for "postacute phase" is >3 months.
OBJECTIVES
The purpose of this study was to determine which cardiovascular diagnoses in the postacute phase of COVID-19 are associated with SARS-CoV-2 infection.
METHODS
Retrospective cohort study of all adults in Alberta who had a positive SARS-CoV-2 reverse transcription polymerase chain reaction test between March 1, 2020 and June 30, 2021, matched (by age, sex, Charlson Comorbidity score, and test date) with controls who had a negative reverse transcription polymerase chain reaction test.
RESULTS
The 177,892 patients with laboratory confirmed SARS-CoV-2 infection (mean age 42.7 years, 49.7% female) were more likely to visit an emergency department (5.7% vs 3.3%), be hospitalized (3.4% vs 2.1%), or die (1.3% vs 0.4%) within 1 month than matched test-negative controls. After 3 months, cases were significantly more likely than controls to have an emergency department visit or hospitalization for diabetes mellitus (1.5% vs 0.7%), hypertension (0.6% vs 0.4%), heart failure (0.2% vs 0.1%), or kidney injury (0.3% vs 0.2%). In the 6,030 patients who had survived a hospitalization for COVID-19, postacute phase risks were substantially greater for diabetes mellitus (9.5% vs 3.0%, adjusted odds ratio [aOR]: 3.16 [95% CI: 2.43-4.12]), hypertension (3.5% vs 1.4%, aOR: 2.89 [95% CI: 1.97-4.23]), heart failure (2.1% vs 0.7%, aOR: 3.16 [95% CI: 1.88-5.29]), kidney injury (3.1% vs 0.8%, aOR: 2.70 [95% CI: 1.71-4.28]), bleeding (1.5% vs 0.5%, aOR: 3.40 [95% CI: 1.83-6.32]), and venous thromboembolism (0.8% vs 0.3%, aOR: 3.60 [95% CI: 1.59-8.13]).
CONCLUSIONS
Clinicians should screen COVID-19 survivors for diabetes mellitus, hypertension, heart failure, and kidney dysfunction in the postacute phase.
PubMed: 38939433
DOI: 10.1016/j.jacadv.2023.100391 -
Cureus May 2024Gastroenteritis is a common cause of morbidity and mortality globally. Its cause encompasses a spectrum of agents, including viruses, bacteria, parasites, toxins, and...
BACKGROUND
Gastroenteritis is a common cause of morbidity and mortality globally. Its cause encompasses a spectrum of agents, including viruses, bacteria, parasites, toxins, and drugs. Viruses account for a considerable portion of gastroenteritis cases across all age groups, typically presenting with symptoms like nausea, vomiting, diarrhea, dehydration, anorexia, and weight loss. While sporadic cases occur, viral gastroenteritis is more frequently observed in outbreaks within closely knit communities such as daycare facilities, nursing homes, and cruise ships. Therefore, it becomes necessary to determine when healthcare providers should consider this condition in their differential diagnosis and to develop the most effective strategy to confirm the diagnosis.
METHODS
De-identified data of patients with gastroenteritis were collected over a five-year period utilizing the Patient Cohort Explorer, an electronic health record at the University of Mississippi Medical Center. Confirmatory laboratory tests employed the BioFire® FilmArray® multiplex polymerase chain reaction for gastrointestinal pathogens. Out of the 22 most common agents associated with gastroenteritis, only viral pathogens, specifically adenovirus, astrovirus, norovirus, rotavirus, and sapovirus, were included in the analysis. When available, histopathology was reviewed.
RESULTS
Among the various causes of gastroenteritis, both infectious and non-infectious, our findings revealed that 25.46% of the cases were linked to viral pathogens. This included a significantly higher percentage of pediatric patients (72.73%) when compared to adults (27.07%), with a p-value of 0.015. Norovirus genogroups I and II emerged as the most frequently detected viruses across all age groups, with a significant prevalence among adults. No discernible gender-based differences were observed. The histopathological findings included inflammation, ulceration, erosion, architectural distortion, and the pathognomonic viral inclusion bodies associated with adenovirus.
CONCLUSION
Our comprehensive analysis of viral gastroenteritis cases highlights the substantial burden of this condition, particularly among pediatric patients. Norovirus emerges as a prevalent culprit which emphasizes the importance of vigilant surveillance and timely diagnosis, especially in settings where outbreaks are common.
PubMed: 38939260
DOI: 10.7759/cureus.61197 -
Cureus May 2024is a new genus of anaerobic bacteria discovered in the year 2000. It is one of the most common bacteria present in the host microbial flora of dental plaque. The levels...
Comparative Evaluation of Mogibacterium timidum in the Subgingival Plaque of Periodontally Healthy and Chronic Periodontitis Patients: A Real-Time Polymerase Chain Reaction (PCR) Study.
BACKGROUND
is a new genus of anaerobic bacteria discovered in the year 2000. It is one of the most common bacteria present in the host microbial flora of dental plaque. The levels of are supposedly higher in inflammatory conditions.
AIMS AND OBJECTIVES
This study aimed to quantify the levels of species in the subgingival plaque samples of healthy patients and patients with chronic periodontitis.
MATERIALS AND METHODS
A total of 24 samples of the subgingival plaque, 12 healthy samples and 12 samples of chronic periodontitis patients, were collected in a buffer solution using a sterile Gracey curette. These samples were then sent to a laboratory for real-time polymerase chain reaction (PCR) testing.
RESULTS
was found in higher quantities in plaque samples taken from chronic periodontitis patients when compared to healthy patients.
CONCLUSION
can be said to be associated with chronic periodontitis condition. Further studies are required to know the exact nature of the pathogen.
PubMed: 38939247
DOI: 10.7759/cureus.61211