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1.
Curr Microbiol ; 80(1): 12, 2022 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-36459233

RESUMO

This study aimed to increase cordycepin production by over-expressing bio-synthetic enzyme genes, including the adenylosuccinate synthase, adenylosuccinate lyase, and 5'-nucleotidase genes. Research data showed that the extracellular and intracellular cordycepin concent of 24 recombinant strains were higher than those of C. militaris WT, indicating that over-expression of key enzyme genes increased cordycepin production. Among them, the CM-adss-5 strain had highest cordycepin production, and the extracellular and intracellular cordycepin concent were 1119.75 ± 1.61 and 65.56 ± 0.97 mg/L, which were 1.26 and 2.61 times that of C. militaris WT. This study also optimized the culture conditions of CM-adss-5 strain through single factor experiments to obtain the best culture conditions. The best culture condition was 25 °C constant temperature, 180-rpm shaking culture, fermentation period 12 days, inoculate amount 5%, initial pH 6, seed age 108 h, and liquid volume 110/250 mL. Then, the extracellular and intracellular cordycepin content of CM-adss-5 strain reached 2581.96 ± 21.07 and 164.08 ± 1.44 mg/L, which were higher by 130.6% and 150.3%, respectively. Therefore, our research provides a way to efficiently produce cordycepin for the development of cordycepin and its downstream products.


Assuntos
Desoxiadenosinas , Sementes , Fermentação , Temperatura
2.
Genome Biol ; 23(1): 249, 2022 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-36461076

RESUMO

BACKGROUND: DNA N6-methyldeoxyadenosine (6mA) is rarely present in mammalian cells and its nuclear role remains elusive. RESULTS: Here we show that hypoxia induces nuclear 6mA modification through a DNA methyltransferase, METTL4, in hypoxia-induced epithelial-mesenchymal transition (EMT) and tumor metastasis. Co-expression of METTL4 and 6mA represents a prognosis marker for upper tract urothelial cancer patients. By RNA sequencing and 6mA chromatin immunoprecipitation-exonuclease digestion followed by sequencing, we identify lncRNA RP11-390F4.3 and one novel HIF-1α co-activator, ZMIZ1, that are co-regulated by hypoxia and METTL4. Other genes involved in hypoxia-mediated phenotypes are also regulated by 6mA modification. Quantitative chromatin isolation by RNA purification assay shows the occupancy of lncRNA RP11-390F4.3 on the promoters of multiple EMT regulators, indicating lncRNA-chromatin interaction. Knockdown of lncRNA RP11-390F4.3 abolishes METTL4-mediated tumor metastasis. We demonstrate that ZMIZ1 is an essential co-activator of HIF-1α. CONCLUSIONS: We show that hypoxia results in enriched 6mA levels in mammalian tumor cells through METTL4. This METTL4-mediated nuclear 6mA deposition induces tumor metastasis through activating multiple metastasis-inducing genes. METTL4 is characterized as a potential therapeutic target in hypoxic tumors.


Assuntos
RNA Longo não Codificante , Neoplasias da Bexiga Urinária , Animais , Metilação , RNA Longo não Codificante/genética , Cromatina , Hipóxia , Desoxiadenosinas , Mamíferos
3.
Molecules ; 27(23)2022 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-36500262

RESUMO

Cordyceps spp. (belonging to the Ascomycota group) are entomopathogenic mushrooms that have traditionally been used in ethnomedicine in Asian countries such as China, Japan, Korea, and India. They are unique parasites of larvae of selected species of moths. Cordyceps militaris is one of the best sources of cordycepin. Worldwide, osteoporosis is one of the most common bone diseases, whose pharmacotherapy includes various medical interventions; however, the research and development of new molecules and new drugs is required. The impact of adenosine receptors (ARs) on the purinergic signaling pathway may regulate proliferation, differentiate dental pulp stem cells and bone marrow, and modulate osteogenesis and bone repair. The aim of the review was to collect and analyze the available data on the effects of Cordyceps spp. or cordycepin on bone function and related processes. To the best of our knowledge, this is the first systematic review in this perspective, not necessarily using mushroom raw material or even the isolated parent compound cordycepin, but new molecules that are analogs of nucleosides, such as those from C. militaris. This review found that Cordyceps spp. or isolated cordycepin interacts via the AR, 5' adenosine monophosphate-activated protein kinase (AMPK), and adenosine-5'-triphosphate (ATP) signaling pathway and evaluated their impact on bones, teeth, and dental pulp. Cordyceps spp. was found to have the potential to develop regenerative medicines, thus providing an opportunity to expand the treatment or intervention methods in the recovery after traumatic injuries, convalescence, and terminal-stage or devastating diseases.


Assuntos
Cordyceps , Osteoporose , Cordyceps/metabolismo , Desoxiadenosinas/farmacologia , Desoxiadenosinas/metabolismo , Transdução de Sinais , China , Ásia
4.
Biochem Biophys Res Commun ; 637: 127-135, 2022 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-36399798

RESUMO

Fatigue, a most commonly sub-health condition, may cause people more susceptible to many diseases. Cordycepin, a principal active ingredient from Cordyceps militaris, exerts various pharmacological activities including anti-diabetes, anti-inflammatory, immunomodulatory and antioxidant effects. However, the anti-fatigue effect of cordycepin and specific mechanism remained unclear. This study aimed to investigate the beneficial effect of cordycepin on physical fatigue and elucidate the potential mechanism. 20 mg/kg, 40 mg/kg of cordycepin and 500 mg/kg taurine were respectively treated to mice for 28 days before weight-loaded swimming test. The results revealed that cordycepin significantly prolonged the weight-loaded swimming time of mice. Meanwhile, cordycepin decreased the levels of lactic acid, blood uric nitrogen, and malondialdehyde, and increased the contents of superoxide dismutase, glutathione, nicotinamide adenine dinucleotide phosphate, hepatic glycogen, muscle glycogen and ATP. The metabolomic study by GC-MS showed that eight biomarkers were found in livers, including L-lactic acid, L-asparagine, 3-phosphoglyceric acid, inosine, D-galactose, L-tyrosine, glyceric acid and L-threonine. There were seven biomarkers in gastrocnemius, including D-ribose-5-phosphate, acetic acid, propionic acid, butyric acid, palmitic acid, oxaloacetic acid and citric acid. The results of metabolomics indicated that cordycepin might relieve fatigue by regulating energy metabolism and pentose phosphate pathway. Furthermore, we found cordycepin significantly enhanced the protein levels of TIGAR, SIRT1, PGC-1α, NRF1 and TFAM in gastrocnemius of weight-loaded swimming mice. Taken together, the present study demonstrated that cordycepin possessed an anti-fatigue effect via activating TIGAR/SIRT1/PGC-1α signaling pathway. Our study indicated that cordycepin may be a potentially efficient candidate for fatigue.


Assuntos
Desoxiadenosinas , Sirtuína 1 , Camundongos , Animais , Desoxiadenosinas/farmacologia , Desoxiadenosinas/uso terapêutico , Transdução de Sinais , Ácido Butírico , Monoéster Fosfórico Hidrolases , Proteínas Reguladoras de Apoptose
5.
Nat Commun ; 13(1): 6138, 2022 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-36253381

RESUMO

Poly-ADP-ribosylation (PARylation) is regarded as a protein-specific modification. However, some PARPs were recently shown to modify DNA termini in vitro. Here, we use ultrasensitive mass spectrometry (LC-MS/MS), anti-PAR antibodies, and anti-PAR reagents to show that mammalian DNA is physiologically PARylated and to different levels in primary tissues. Inhibition of PAR glycohydrolase (PARG) increases DNA PARylation, supporting that the modification is reversible. DNA PARylation requires PARP1 and in vitro PARP1 PARylates single-stranded DNA, while PARG reverts the modification. DNA PARylation occurs at the N1-position of adenosine residues to form N1-Poly(ADP-ribosyl)-deoxyadenosine. Through partial hydrolysis of mammalian gDNA we identify PAR-DNA via the diagnostic deamination product N1-ribosyl-deoxyinosine to occur in vivo. The discovery of N1-adenosine PARylation as a DNA modification establishes the conceptual and methodological framework to elucidate its biological relevance and extends the role of PARP enzymes.


Assuntos
Poli ADP Ribosilação , Inibidores de Poli(ADP-Ribose) Polimerases , Adenosina , Difosfato de Adenosina , Animais , Cromatografia Líquida , DNA/metabolismo , DNA de Cadeia Simples , Desoxiadenosinas , Glicosídeo Hidrolases/metabolismo , Mamíferos/genética , Poli(ADP-Ribose) Polimerase-1/metabolismo , Poli Adenosina Difosfato Ribose/metabolismo , Espectrometria de Massas em Tandem
6.
Bioelectrochemistry ; 148: 108264, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36122426

RESUMO

Carbon material derived from the waste-based biomass human hair (H), which is naturally rich in pyridinic nitrogen, provides a significant benefit in biosensor applications with its dominant conductivity character. The carbon material was synthesized from human hair waste by the hydrothermal carbonization (HTC) method, which is a promising green synthesis. A morphological characterization of the carbon materials was performed. In this study, H and amine-functionalized multi-walled carbon nanotubes (NH2-MWCNT) were combined for the first time as a modifier, which enhanced the glassy carbon electrode (GCE) surface area for deoxyribonucleic acid (DNA) biosensor studies. Palbociclib (PLB) is clinically used in the treatment of breast cancer. The novel electrochemical nanobiosensor was used to investigate the dsDNA-PLB interaction to evaluate the possibility that PLB causes conformational changes in DNA structure and/or oxidative damage. The interaction was conducted based on the voltammetric signals of deoxyguanosine (dGuo) and deoxyadenosine (dAdo) by differential pulse voltammetry (DPV) on a bare and H + NH2-MWCNT modified GCE. The proposed analytical method was applied to a pharmaceutical dosage form with a satisfactory recovery of 98.25 %. The nanobiosensor was tested in the presence of some interfering agents. The binding mechanism of dsDNA-PLB was also evaluated by spectroscopic and theoretical calculations.


Assuntos
Técnicas Biossensoriais , Nanotubos de Carbono , Aminas , DNA/química , Desoxiadenosinas , Desoxiguanosina , Técnicas Eletroquímicas/métodos , Eletrodos , Cabelo , Humanos , Nanotubos de Carbono/química , Nitrogênio/química , Preparações Farmacêuticas , Piperazinas , Piridinas
7.
Int J Mol Sci ; 23(18)2022 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-36142286

RESUMO

We previously found that cordycepin inhibits the growth and metastasis formation of MDA-MB-231 cells through the Hedgehog pathway but has not validated this in vivo. In this study, we confirmed cordycepin's anti-triple-negative breast cancer (TNBC) effect in nude mice and documented its mechanism. We found that cordycepin reduced the volume and weight of MDA-MB-231 xenografts and affected the expression of proliferation-, apoptosis-, epithelial-mesenchymal transition-, and matrix metalloproteinase-related proteins without side effects. RNA sequencing screening, pathway enrichment, and the protein network interaction analysis revealed enriched pathways and targets mainly concentrated on the Hedgehog pathway and its core components of SHH and GLI2. This indicates that the Hedgehog pathway plays a central role in the cordycepin-mediated regulation of growth and metastasis formation in TNBC. The database analysis of the Hedgehog pathway markers (SHH, PTCH1, SMO, GLI1, and GLI2) revealed that the Hedgehog pathway is activated in breast cancer tissues, and its high expression is not conducive to a patient's survival. Finally, we verified that cordycepin effectively inhibited the Hedgehog pathway in TNBC through Western blotting and immunohistochemistry. This study found that cordycepin could regulate the growth and metastasis formation of TNBC through the Hedgehog pathway in vivo, which provides new insights for targeting and treating breast cancer.


Assuntos
Proteínas Hedgehog , Neoplasias de Mama Triplo Negativas , Animais , Linhagem Celular Tumoral , Proliferação de Células , Desoxiadenosinas , Proteínas Hedgehog/metabolismo , Xenoenxertos , Humanos , Camundongos , Camundongos Nus , Neoplasias de Mama Triplo Negativas/metabolismo , Proteína GLI1 em Dedos de Zinco/genética
8.
Biomed Pharmacother ; 154: 113619, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36081285

RESUMO

Numerous researchers have investigated cordycepin (COR) as an anti-tumor compound. COR has been documented to have cytotoxic effects on several cancer cells. The current work used a Box-Behnken mathematical design to minimize COR's size. The design incorporated COR concentration, phospholipid concentration and sonication time as variables to minimize the vesicles of COR emulsomes (COR-EMLs). To evaluate degree of improvement of COR cytotoxicity against colorectal cancer (HCT116) cells, cell viability, cell cycle analysis and apoptosis have been assessed. In addition, wound scratching and mitochondrial membrane potential were evaluated. Results of Box-Behnken design achieved COR-EMLs sizes in range from 91.54 ± 2.3-343.83 ± 3.7 nm. Moreover, the optimized formula morphology's was evaluated using transmission electron microscope and showed nanospheres in range of 100 nm. COR released from COR-EMLs exceeded 80% after 12 h.The half-maximal inhibitory concentration (IC50) of the refined COR-EML formula was about four times lower than that of COR-raw. The cell cycle study revealed that administration of COR-EML considerably hindered HCT116 cellular propagation in contrast to plain emulsomes (EMLs) and COR-raw with a denser cell compilation in G2/M. Moreover, the optimized formula notably enhanced the proportion of cells in both the initial and late phases of apoptosis. The augmentation of COR cytotoxicity was confirmed by its inhibition of cancer cell wound healing by approximately 40%. The mitochondrial membrane potential was significantly lower than in cells treated with COR-raw and EMLs. Finally, loading COR into the EMLs increased COR's capacity to lower mitochondrial membrane functionality and significantly improved its cytotoxicity.


Assuntos
Neoplasias do Colo , Desoxiadenosinas , Apoptose , Sobrevivência Celular , Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/patologia , Desoxiadenosinas/farmacologia , Células HCT116 , Humanos
9.
Biomed Pharmacother ; 153: 113491, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36076585

RESUMO

Cordyceps militaris is rich in adenosine derivatives, including 3'-deoxyadenosine, also known as cordycepin. It has been reported for antitumor effects, but its underlying molecular mechanism has yet to be elucidated. We investigated how adenosine derivatives exerted antitumor effects against ovarian cancer using human ovarian cancer cells and a xenograft mouse model. Treatment with adenosine derivatives effectively resulted in cell death of ovarian cancer cells through AMPK activation and subsequently mTOR-mediated autophagic induction. Intriguingly, the effect required membrane transport of adenosine derivatives via ENT1, rather than ADORA-mediated cellular signaling. Our data suggest that adenosine derivatives may be an effective therapeutic intervention in ovarian cancer through induction of ENT1-AMPK-mTOR-mediated autophagic cell death.


Assuntos
Adenosina , Morte Celular Autofágica , Cordyceps , Neoplasias Ovarianas , Proteínas Quinases Ativadas por AMP/efeitos dos fármacos , Proteínas Quinases Ativadas por AMP/metabolismo , Adenosina/análogos & derivados , Adenosina/metabolismo , Adenosina/farmacologia , Animais , Morte Celular Autofágica/efeitos dos fármacos , Carcinoma Epitelial do Ovário , Cordyceps/química , Desoxiadenosinas/farmacologia , Transportador Equilibrativo 1 de Nucleosídeo/efeitos dos fármacos , Transportador Equilibrativo 1 de Nucleosídeo/metabolismo , Feminino , Humanos , Camundongos , Neoplasias Ovarianas/tratamento farmacológico , Serina-Treonina Quinases TOR/metabolismo
10.
J Am Chem Soc ; 144(35): 16150-16156, 2022 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-36001794

RESUMO

The formation of unnatural base pairs within duplex DNA would facilitate DNA nanotechnology and biotechnology. Iso-2'-deoxyguanosine (iso-dG) forms base pairs with iso-2'-deoxycytidine, and its use as an unnatural base pair was investigated. Iso-dG is one of the tautomers of 2-hydroxy-2'-deoxyadenosine (2-OH-dA), known as an oxidatively damaged nucleobase, and its selective recognition in DNA plays an important role in the diagnosis and pathogenesis of disease. Therefore, we focused on pseudo-dC (ψdC) as a suitable molecule that recognizes 2-OH-dA in DNA. Since 2-OH-dA shows tautomeric structures in DNA, we designed and used ψdC, which also has a tautomeric structure. We successfully synthesized a ψdC phosphoramidite compound for the synthesis of oligonucleotides (ODNs) as well as its triphosphate derivative (ψdCTP). Tm measurements revealed that ODNs including ψdC showed stable base pair formation with ODNs having 2-OH-dA. In contrast, low Tm values were observed for other bases (dG, dA, dC, and T). The results obtained for the single-nucleotide primer extension reaction revealed that ψdCTP was incorporated into the complementary position of 2-OH-dA in template DNA with high selectivity. In addition, the primer elongation reaction was confirmed to proceed in the presence of dNTPs. The present study reports an artificial nucleic acid that selectively and stably forms unnatural base pairs with 2-OH-dA in DNA.


Assuntos
DNA , Desoxiadenosinas , Pareamento de Bases , DNA/química , Desoxiadenosinas/química , Conformação de Ácido Nucleico , Oligonucleotídeos/química
11.
Int J Med Mushrooms ; 24(8): 1-20, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35997091

RESUMO

The pharmacological values of Cordyceps spp. are substantially associated with the existence of an extremely potent biometabolite: cordycepin. This component exhibits powerful therapeutic activity against cancer, diabetes, and hyperlipidemia and acts as a strong immunomodulator. Extensive pharmaceutical exploitation of Cordyceps spp. has depleted its natural existence. Therefore, there is a strong need for metabolic engineering-based approaches that could be employed for overproduction of the desired metabolite, which would sustain market demands. Replacement of the old conventional genome editing tools by the newly developed CRISPR technology is considered a suitable alternative for enhancing metabolite production. Another novel approach, POPCORN, optimizes carbon/nitrogen ratios to design synthetic media for Cordyceps production. In fact, the addition of FeSO4 and porcine liver extract and alterations in the dissolved oxygen enhanced cordycepin production in the submerged state. Ultraviolet mutagenesis is another approach for the augmentation of this pharmaceutically potent biometabolite. Therefore, the main objective of this review is to present the outlook on pharmaceutical properties of cordycepin along with the metabolic approaches for enhancing cordycepin production.


Assuntos
Cordyceps , Desoxiadenosinas , Animais , Cordyceps/metabolismo , Meios de Cultura/metabolismo , Preparações Farmacêuticas/metabolismo , Suínos
12.
Biochemistry ; 61(17): 1883-1893, 2022 09 06.
Artigo em Inglês | MEDLINE | ID: mdl-35969806

RESUMO

Enzyme-catalyzed hydrolysis is a fundamental chemical transformation involved in many essential metabolic processes. The enzyme 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (MTAN) catalyzes the hydrolysis of adenosine-containing metabolites in cysteine and methionine metabolism. Although MTAN enzymes contain highly similar active site architecture and generally follow a dissociative (DN*AN) reaction mechanism, substantial differences in reaction rates and chemical transition state structures have been reported. To understand how subtle changes in sequence and structure give rise to differences in chemistry between homologous enzymes, we have probed the reaction coordinates of two MTAN enzymes using quantum mechanical/molecular mechanical and molecular dynamics simulations combined with experimental methods. We show that the transition state structure and energy are significantly affected by the recruitment and positioning of the catalytic water molecule and that subtle differences in the noncatalytic active site residues alter the environment of the catalytic water, leading to changes in the reaction coordinate and observed reaction rate.


Assuntos
N-Glicosil Hidrolases , Água , Catálise , Desoxiadenosinas , Hidrólise , N-Glicosil Hidrolases/química , Purina-Núcleosídeo Fosforilase , Tionucleosídeos
13.
J Biol Chem ; 298(9): 102367, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35963436

RESUMO

Methylthioadenosine phosphorylase (MTAP) is a key enzyme in the methionine salvage pathway that converts the polyamine synthesis byproduct 5'-deoxy-5'-methylthioadenosine (MTA) into methionine. Inactivation of MTAP, often by homozygous deletion, is found in both solid and hematologic malignancies and is one of the most frequently observed genetic alterations in human cancer. Previous work established that MTAP-deleted cells accumulate MTA and contain decreased amounts of proteins with symmetric dimethylarginine (sDMA). These findings led to the hypothesis that accumulation of intracellular MTA inhibits the protein arginine methylase (PRMT5) responsible for bulk protein sDMAylation. Here, we confirm that MTAP-deleted cells have increased MTA accumulation and reduced protein sDMAylation. However, we also show that addition of extracellular MTA can cause a dramatic reduction of the steady-state levels of sDMA-containing proteins in MTAP+ cells, even though no sustained increase in intracellular MTA is found because of catabolism of MTA by MTAP. We determined that inhibition of protein sDMAylation by MTA occurs within 48 h, is reversible, and is specific. In addition, we have identified two enhancer-binding proteins, FUBP1 and FUBP3, that are differentially sDMAylated in response to MTAP and MTA. These proteins work via the far upstream element site located upstream of Myc and other promoters. Using a transcription reporter construct containing the far upstream element site, we demonstrate that MTA addition can reduce transcription, suggesting that the reduction in FUBP1 and FUBP3 sDMAylation has functional consequences. Overall, our findings show that extracellular MTA can inhibit protein sDMAylation and that this inhibition can affect FUBP function.


Assuntos
Arginina , Desoxiadenosinas , Purina-Núcleosídeo Fosforilase , Arginina/análogos & derivados , Proteínas de Ligação a DNA/metabolismo , Humanos , Metionina/metabolismo , Metilação , Poliaminas , Proteína-Arginina N-Metiltransferases/genética , Proteína-Arginina N-Metiltransferases/metabolismo , Purina-Núcleosídeo Fosforilase/genética , Purina-Núcleosídeo Fosforilase/metabolismo , Proteínas de Ligação a RNA/metabolismo , Deleção de Sequência , Tionucleosídeos
14.
J Acquir Immune Defic Syndr ; 91(1): 68-72, 2022 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-35972855

RESUMO

BACKGROUND: Islatravir (MK-8591) is a nucleoside reverse transcriptase translocation inhibitor in development for treatment and prevention of HIV-1. We present efficacy and safety data for islatravir and doravirine (DOR) through 96 weeks of the phase 2b trial (NCT03272347). METHODS: In this randomized, double-blind, dose-ranging trial, participants initially received islatravir (0.25, 0.75, or 2.25 mg) with doravirine (100 mg) and lamivudine (3TC, 300 mg) or a fixed-dose combination of doravirine, 3TC, and tenofovir disoproxil fumarate (DOR/3TC/TDF) daily. Beginning at week 24, participants receiving islatravir stopped 3TC if HIV-1 RNA from the prior visit was <50 copies per milliliter and continued taking the assigned islatravir dose (still blinded) with doravirine. All islatravir groups transitioned to open-label use of 0.75 mg between weeks 60 and 84. Efficacy end points at week 96 included the proportion of participants maintaining HIV-1 RNA of <50 copies per milliliter (FDA Snapshot). Safety was assessed by adverse event (AE) reporting. RESULTS: One hundred twenty-one treatment-naive participants received the study drugs and were included in the analyses. Through week 96, HIV-1 RNA<50 copies per milliliter was maintained in 86.2% (25/29), 90.0% (27/30), and 67.7% (21/31) of participants in the 0.25-, 0.75-, and 2.25-mg islatravir groups, respectively, 81.1% (73/90) of the combined islatravir group, and 80.6% (25/31) of the DOR/3TC/TDF group. One participant in the 2.25-mg islatravir group had Protocol-Defined Virologic Failure after week 48. Drug-related AE rates were higher for DOR/3TC/TDF participants (22.6%) compared with islatravir (combined 7.8%). Two participants (2.2%) receiving islatravir with doravirine and one (3.2%) receiving DOR/3TC/TDF discontinued because of an AE. CONCLUSIONS: Treatment regimens containing islatravir and doravirine maintained viral suppression through week 96 and were well tolerated regardless of dose.


Assuntos
Desoxiadenosinas , Infecções por HIV , HIV-1 , Adulto , Fármacos Anti-HIV/uso terapêutico , Desoxiadenosinas/administração & dosagem , Desoxiadenosinas/efeitos adversos , Combinação de Medicamentos , Infecções por HIV/tratamento farmacológico , Humanos , Lamivudina/uso terapêutico , Piridonas/uso terapêutico , RNA , Inibidores da Transcriptase Reversa/efeitos adversos , Tenofovir/uso terapêutico , Triazóis
15.
Anal Chem ; 94(32): 11248-11254, 2022 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-35924299

RESUMO

DNA methylation can occur naturally or be induced by various environmental and chemotherapeutic agents. The regioisomeric N1- and N6-methyldeoxyadenosine (1mdA and 6mdA, respectively) represent an important class of methylated DNA adducts. In this study, we developed a shuttle vector- and next-generation sequencing-based assay to quantitatively assess the effects of 1mdA and 6mdA on the accuracy and efficiency of DNA transcription. Our results revealed that 1mdA can induce multiple types of mutant transcripts and strongly inhibit DNA transcription, whereas 6mdA is a nonmutagenic DNA adduct that can exhibit a subtle but significant inhibitory effect on DNA transcription in vitro and in human cells. Moreover, our results demonstrated that the transcription-coupled nucleotide excision repair pathway is dispensable for the removal of 1mdA and 6mdA from the template DNA strand in human cells. These findings provided new important insights into the functional interplay between DNA methylation modifications and transcription in mammalian cells.


Assuntos
Adutos de DNA , Transcrição Genética , Animais , DNA/genética , DNA/metabolismo , Reparo do DNA , Desoxiadenosinas , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Mamíferos/metabolismo
16.
Bioresour Technol ; 363: 127862, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36041680

RESUMO

Cordycepin is a nucleoside antibiotic with various biological activities, which has wide applications in the area of cosmetic and medicine industries. However, the current production of cordycepin is costly and time-consuming. To construct the promising cell factory for high-level cordycepin production, firstly, the design and construction of cordycepin biosynthetic pathway were performed in Yarrowia lipolytica. Secondly, the adaptivity between cordycepin biosynthetic pathway and Y. lipolytica was enhanced by enzyme fusion and integration site engineering. Then, the production of cordycepin was improved by the enhancement of adenosine supply. Furthermore, through modular engineering, the production of cordycepin was achieved at 3588.59 mg/L from glucose. Finally, 3249.58 mg/L cordycepin with a yield of 76.46 mg/g total sugar was produced by the engineered strain from the mixtures of glucose and molasses. This research is the first report on the de novo high-level production of cordycepin in the engineered Y. lipolytica.


Assuntos
Yarrowia , Adenosina/metabolismo , Antibacterianos/metabolismo , Desoxiadenosinas , Glucose/metabolismo , Engenharia Metabólica , Nucleosídeos , Açúcares/metabolismo , Yarrowia/genética , Yarrowia/metabolismo
17.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 38(6): 513-521, 2022 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-35732609

RESUMO

Objective To explore the inhibitory effect of cordycepin on the proliferation and migration of gastric cancer cells and its molecular mechanism. Methods MGC-803 cells were treated with 0, 25, 50, 100 µmol/L of cordycepin and HGC-27 cells with 0, 5, 25, 50 µmol/L of cordycepin for 48 hours. The proliferation ability of MGC-803 and HGC-27 cells was detected by MTT assay and EdU assay; the colony formation ability of cells was detected by colony formation assay; both wound healing assay and cell migration assay were applied to detect the cell migration ability of MGC-803 and HGC-27 cells; the chromatin agglutination was detected by Hoechst 33342 staining; the apoptosis of gastric cancer cells was detected by annexin V-FITC/PI double labeling combined with flow cytometry; Western blot was used to measure the protein expression levels of lipid metabolism-related proteins including sterol regulatory element binding transcription factor 1 (SREBF1), fatty acid synthase (FASN), and acetyl coA carboxylase 1 (ACC1), epithelial-mesenchymal transition (EMT)-related proteins including E-cadherin, vimentin, Snail, Slug, matrix metalloproteinase 2 (MMP2), MMP9, AMPK, and phosphorylated AMPK (p-AMPK), MAPK signaling pathway-related proteins including JNK, phosphorylated JNK (p-JNK), p38 MAPK, and p-p38 MAPK, and apoptosis-related proteins including cleaved caspase-9 (c-caspase-9), c-caspase-3, and cleaved poly (ADP-ribose) polymerase (c-PARP). Results Cordycepin significantly inhibited the proliferation, colony formation, and migration of gastric cancer cells. After cordycepin treatment, the karyopycnosis, karyorrhexis, and apoptosis rate of cancer cells increased, and the expressions of apoptosis-related proteins c-caspase-3, c-caspase-9, and c-PARP increased. The expression of E-cadherin increased, while the expressions of vimentin, Snail, Slug, SREBF1, FASN, ACC1, MMP2, MMP9 significantly decreased; the phosphorylation levels of AMPK, JNK and P38 proteins significantly increased. Conclusion Cordycepin inhibits the proliferation and migration of gastric cancer cells by suppressing the lipid metabolism and EMT process via activating AMPK and MAPK signaling pathway.


Assuntos
Desoxiadenosinas , Neoplasias Gástricas , Proteínas Quinases Ativadas por AMP/metabolismo , Apoptose , Caderinas/metabolismo , Caspase 3/metabolismo , Caspase 9/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Desoxiadenosinas/farmacologia , Humanos , Metabolismo dos Lipídeos , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Inibidores de Poli(ADP-Ribose) Polimerases/farmacologia , Inibidores de Poli(ADP-Ribose) Polimerases/uso terapêutico , Neoplasias Gástricas/tratamento farmacológico , Neoplasias Gástricas/metabolismo , Vimentina/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
18.
Folia Microbiol (Praha) ; 67(6): 851-860, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35678982

RESUMO

Cordycepin is an essential nucleoside antibiotic with a broad spectrum of physiological functions, which is currently produced by the fermentation of Cordyceps militaris. Even though numerous efforts were made to enhance cordycepin production, the cordycepin yield is still limited. High-cordycepin-yielding strains are still a prerequisite for industrial cordycepin production in large amounts. Screening high-cordycepin-yielding strains from other sources may break new grounds for cordycepin. In this study, Cordyceps hawkesii Gray, with high homology to C. militaris, was selected as the source to screen the cordycepin manufacturing endophytic fungi. Four isolates capable of cordycepin production were successfully obtained among all isolated endophytic fungi. One of the four with better cordycepin yield was identified as Irpex lacteus CHG05, which belongs to the Phlebia species. The response surface methodology was applied to optimize the culture conditions for cordycepin fermentation. 162.05 mg/L of cordycepin with a 53.1% improvement was achieved compared to the original conditions. This study indicates that the endophytic fungi from C. hawkesii Gray could produce cordycepin and served as the first report for cordycepin by the white-rot fungus of I. lacteus. Even though the yield is low compared to C. militaris, this strain provided another choice for enhanced cordycepin in the future.


Assuntos
Cordyceps , Desoxiadenosinas
19.
Curr Opin HIV AIDS ; 17(4): 240-246, 2022 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-35762379

RESUMO

PURPOSE OF REVIEW: To summarize recent updates on the potential role of islatravir for HIV treatment and prevention. RECENT FINDINGS: Islatravir is an investigational antiretroviral agent with unique pharmacologic properties that facilitate flexible dosing regimens. Islatravir has demonstrated potent antiviral activity and a high barrier to resistance when combined with doravirine and lamivudine. A simplified two-drug HIV treatment regimen of islatravir combined with doravirine has also demonstrated comparable efficacy to standard of care three-drug regimens. The long half-life and high potency of islatravir's active metabolite may support its use as a long-acting option for HIV preexposure prophylaxis (PrEP). A once monthly oral dose of islatravir maintains effective concentrations of its active metabolite over the entire dosing interval. Furthermore, an investigational implantable formulation has been projected to provide efficacious concentrations for at least a year and exhibits comparable distribution into vaginal and rectal tissues making it a promising PrEP option for male and female individuals. Islatravir has minimal risks of drug interactions as it is not a substrate, inducer, or inhibitor of major drug metabolizers and transporters. Finally, clinical trials demonstrate islatravir's favorable safety profile revealing only mild and transient adverse events. SUMMARY: Leveraging the unique pharmacological properties of islatravir offers opportunities for simplified HIV treatment regimens and long-acting PrEP making it a valuable addition to the antiretroviral arsenal.


Assuntos
Fármacos Anti-HIV , Infecções por HIV , HIV-1 , Profilaxia Pré-Exposição , Desoxiadenosinas , Feminino , Infecções por HIV/tratamento farmacológico , Infecções por HIV/prevenção & controle , Humanos , Masculino
20.
Mol Biol Rep ; 49(9): 8673-8683, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35763180

RESUMO

BACKGROUND: Hyperthermia induces cancer cell death. However, the cytotoxic effect of hyperthermia is not sufficient. Cordycepin can also induce apoptosis in cancer cells and enhance the antitumoral activity of irradiation. To examine cordycepin-mediated enhancement of hyperthermia-induced apoptosis, this study investigated the combined effects and apoptotic mechanisms of hyperthermia and cordycepin on human leukemia U937 cells. METHODS: Cell viability and apoptosis were measured using MTT assays, Hoechst 33342 staining and Annexin V/PI double staining. The distribution of the cell cycle and sub-G1 phase, reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) were examined by flow cytometry. The expression of related proteins was analyzed by western blotting. RESULTS: Combined treatment with hyperthermia and cordycepin markedly augmented apoptosis by upregulating Bax and suppressing Bcl-2, Bid and activated caspase 3 and 8 expression, and apoptosis was decreased by Z-VAD-fmk (a pan caspase inhibitor). We also found that the MMP was significantly decreased and excessive ROS generation occurred. The combination treatment also induced arrest in the G2/M phase by downregulating cyclin dependent kinase 1 (CDK1) and cyclin B1 protein expression. Furthermore, it was observed that mitogen-activated protein kinase (MAPK) pathway including ERK, JNK and p38 signals was involved in the induction of apoptosis. The phosphorylated p38 and JNK were increased and ERK phosphorylation was decreased by the combined treatment. In addition, N-acetyl-L-cysteine (NAC) significantly protected the cells by restoring ROS levels and the activity of caspase-3, inactivating the MAPK pathway. CONCLUSION: Cordycepin significantly enhanced hyperthermia-induced apoptosis and G2/M phase arrest in U937 cells. The combined treatment enhanced apoptosis through the MAPK pathway and mitochondrial dysfunction, and these effects could be rescued by NAC. We report for the first time that cordycepin can be used as a hyperthermia sensitizer to treat leukemia.


Assuntos
Hipertermia Induzida , Leucemia , Linfoma , Apoptose , Pontos de Checagem do Ciclo Celular , Linhagem Celular Tumoral , Desoxiadenosinas , Humanos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Células U937 , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
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