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2.
Metabolites ; 14(3)2024 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-38535303

RESUMO

In vitro fertilization (IVF) is a highly effective treatment for infertility; however, it poses challenges for women with decreased ovarian reserve (DOR). Despite the importance of understanding the impact of DOR on IVF outcomes, limited research has explored this relationship, particularly using omics approaches. Hence, we conducted a study to investigate the association between DOR and IVF outcomes, employing a metabolomic approach. We analyzed serum samples from 207 women undergoing IVF treatment, including 89 with DOR and 118 with normal ovarian reserve (NOR). Our findings revealed that DOR was significantly associated with unfavorable IVF outcomes, characterized by a reduced oocyte count, lower embryo quality, and decreased rates of pregnancy and live births. Furthermore, we identified 82 metabolites that displayed significant alterations in DOR patients, impacting diverse metabolic pathways. Notably, a distinct panel of metabolites, including palmitic acid, stearic acid, LysoPC(9:0(CHO)/0:0), PC(18:0/9:0(CHO)), and PC(16:0/9:0(CHO)), exhibited discriminatory power between the DOR and NOR groups, showcasing a strong correlation with IVF outcomes. These findings emphasize the crucial role of metabolomic disruptions in influencing IVF outcomes among women with DOR.

3.
Se Pu ; 42(2): 159-163, 2024 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-38374596

RESUMO

Peak alignment is a crucial data-processing step in untargeted metabolomics analysis that aims to integrate metabolite data from multiple liquid chromatography-mass spectrometry (LC-MS) batches for enhanced comparability and reliability. However, slight variations in the chromatographic separation conditions can result in retention time (RT) shifts between consecutive analyses, adversely affecting peak alignment accuracy. In this study, we present a retention index (RI)-based chromatographic peak-shift correction (CPSC) strategy to address RT shifts and align chromatographic peaks for metabolomics studies. A series of N-acyl glycine homologues (C2-C23) was synthesized as calibrants, and an LC RI system was established. This system effectively corrected RT shifts arising from variations in flow rate, gradient elution, instrument systems, and chromatographic columns. Leveraging the RI system, we successfully adjusted the RT of raw data to mitigate RT shifts and then implemented the Joint Aligner algorithm for peak alignment. We assessed the accuracy of the RI-based CPSC strategy using pooled human fecal samples as a test model. Notably, the application of the RI-based CPSC strategy to a long-term dataset spanning 157 d as an illustration revealed a significant enhancement in peak alignment accuracy from 15.5% to 80.9%, indicating its ability to substantially improve peak-alignment precision in multibatch LC-MS analyses.


Assuntos
Algoritmos , Metabolômica , Humanos , Reprodutibilidade dos Testes , Cromatografia Líquida , Espectrometria de Massa com Cromatografia Líquida
5.
Anal Chem ; 95(35): 13330-13337, 2023 09 05.
Artigo em Inglês | MEDLINE | ID: mdl-37609864

RESUMO

Peak alignment is a crucial step in liquid chromatography-mass spectrometry (LC-MS)-based large-scale untargeted metabolomics workflows, as it enables the integration of metabolite peaks across multiple samples, which is essential for accurate data interpretation. Slight differences or fluctuations in chromatographic separation conditions, however, can cause the chromatographic retention time (RT) shift between consecutive analyses, ultimately affecting the accuracy of peak alignment between samples. Here, we introduce a novel RT shift correction method based on the retention index (RI) and apply it to peak alignment. We synthesized a series of N-acyl glycine (C2-C23) homologues via the amidation reaction between glycine with normal saturated fatty acids (C2-C23) as calibrants able to respond proficiently in both mass spectrometric positive- and negative-ion modes. Using these calibrants, we established an N-acyl glycine RI system. This RI system is capable of covering a broad chromatographic space and addressing chromatographic RT shift caused by variations in flow rate, gradient elution, instrument systems, and LC separation columns. Moreover, based on the RI system, we developed a peak shift correction model to enhance peak alignment accuracy. Applying the model resulted in a significant improvement in the accuracy of peak alignment from 15.5 to 80.9% across long-term data spanning a period of 157 days. To facilitate practical application, we developed a Python-based program, which is freely available at https://github.com/WHU-Fenglab/RI-based-CPSC.


Assuntos
Fabaceae , Cromatografia Líquida , Glicina , Espectrometria de Massas , Metabolômica
6.
Anal Chem ; 95(30): 11550-11557, 2023 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-37471289

RESUMO

Gut microbiota-host co-metabolites serve as essential mediators of communication between the host and gut microbiota. They provide nutrient sources for host cells and regulate gut microenvironment, which are associated with a variety of diseases. Analysis of gut microbiota-host co-metabolites is of great significance to explore the host-gut microbiota interaction. In this study, we integrated chemical derivatization, liquid chromatography-mass spectrometry, and molecular networking (MN) to establish a novel CD-MN strategy for the analysis of carboxylated metabolites in gut microbial-host co-metabolism. Using this strategy, 261 carboxylated metabolites from mouse feces were detected, which grouped to various classes including fatty acids, bile acids, N-acyl amino acids, benzoheterocyclic acids, aromatic acids, and other unknown small-scale molecular clusters in MN. Based on the interpretation of the bile acid cluster, a novel type of phenylacetylated conjugates of host bile acids was identified, which were mediated by gut microbiota and exhibited a strong binding ability to Farnesoid X receptor and Takeda G protein-coupled receptor 5. Our proposed strategy offers a promising platform for uncovering carboxylated metabolites in gut microbial-host co-metabolism.


Assuntos
Microbioma Gastrointestinal , Animais , Camundongos , Microbioma Gastrointestinal/fisiologia , Metaboloma , Fezes/química , Espectrometria de Massas/métodos , Aminoácidos/análise , Ácidos e Sais Biliares/análise
8.
J Chromatogr A ; 1702: 464083, 2023 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-37230052

RESUMO

Central carbon metabolism pathway (CCM) is one of the most important metabolic pathways in all living organisms and play crucial function in aspect of organism life. However, the simultaneous detection of CCM intermediates remains challenging. Here, we developed a chemical isotope labeling combined with LC-MS method for simultaneous determination of CCM intermediates with high coverage and accuracy. By chemical derivatization with 2-(diazo-methyl)-N-methyl-N-phenyl-benzamide (2-DMBA) and d5-2-DMBA, all CCM intermediates obtain better separation and accurate quantification at a single LC-MS run. The obtained limits of detection of CCM intermediates ranged from 5 to 36 pg/mL. Using this method, we achieved simultaneous and accurate quantification of 22 CCM intermediates in different biological samples. Take account of the high detection sensitivity of the developed method, this method was further applied to the quantification of CCM intermediates at single-cell level. Finally, 21 CCM intermediates were detected in 1000 HEK-293T cells and 9 CCM intermediates were detected in mouse kidney glomeruli optical slice samples (10∼100 cells).


Assuntos
Carbono , Camundongos , Animais , Carbono/metabolismo , Marcação por Isótopo/métodos , Espectrometria de Massas , Cromatografia Líquida/métodos
9.
Plant Commun ; 4(4): 100567, 2023 07 10.
Artigo em Inglês | MEDLINE | ID: mdl-36855304

RESUMO

Wheat (Triticum aestivum, BBAADD) is an allohexaploid species that originated from two polyploidization events. The progenitors of the A and D subgenomes have been identified as Triticum urartu and Aegilops tauschii, respectively. Current research suggests that Aegilops speltoides is the closest but not the direct ancestor of the B subgenome. However, whether Ae. speltoides has contributed genomically to the wheat B subgenome and which chromosome regions are conserved between Ae. speltoides and the B subgenome remain unclear. Here, we assembled a high-quality reference genome for Ae. speltoides, resequenced 53 accessions from seven species (Aegilops bicornis, Aegilops longissima, Aegilops searsii, Aegilops sharonensis, Ae. speltoides, Aegilops mutica [syn. Amblyopyrum muticum], and Triticum dicoccoides) and revealed their genomic contributions to the wheat B subgenome. Our results showed that centromeric regions were particularly conserved between Aegilops and Triticum and revealed 0.17 Gb of conserved blocks between Ae. speltoides and the B subgenome. We classified five groups of conserved and non-conserved genes between Aegilops and Triticum, revealing their biological characteristics, differentiation in gene expression patterns, and collinear relationships between Ae. speltoides and the wheat B subgenome. We also identified gene families that expanded in Ae. speltoides during its evolution and 789 genes specific to Ae. speltoides. These genes can serve as genetic resources for improvement of adaptability to biotic and abiotic stress. The newly constructed reference genome and large-scale resequencing data for Sitopsis species will provide a valuable genomic resource for wheat genetic improvement and genomic studies.


Assuntos
Pão , Triticum , Triticum/genética , Genoma de Planta/genética , Mapeamento Cromossômico , Poaceae/genética
10.
Nat Commun ; 14(1): 836, 2023 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-36788238

RESUMO

Wheat (Triticum aestivum L.) is a major staple food for more than one-third of the world's population. Tiller number is an important agronomic trait in wheat, but only few related genes have been cloned. Here, we isolate a wheat mutant, tiller number1 (tn1), with much fewer tillers. We clone the TN1 gene via map-based cloning: TN1 encodes an ankyrin repeat protein with a transmembrane domain (ANK-TM). We show that a single amino acid substitution in the third conserved ankyrin repeat domain causes the decreased tiller number of tn1 mutant plants. Resequencing and haplotype analysis indicate that TN1 is conserved in wheat landraces and modern cultivars. Further, we reveal that the expression level of the abscisic acid (ABA) biosynthetic gene TaNCED3 and ABA content are significantly increased in the shoot base and tiller bud of the tn1 mutants; TN1 but not tn1 could inhibit the binding of TaPYL to TaPP2C via direct interaction with TaPYL. Taken together, we clone a key wheat tiller number regulatory gene TN1, which promotes tiller bud outgrowth probably through inhibiting ABA biosynthesis and signaling.


Assuntos
Pão , Triticum , Triticum/genética , Repetição de Anquirina/genética , Fenótipo
11.
J Am Coll Health ; 71(5): 1407-1416, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-34242538

RESUMO

OBJECTIVE: Changes in surroundings and social relationships may heighten feelings of loneliness, suggesting the need to measure as a state. This study tested whether loneliness fluctuates within and across days and the resultant associations with psychological distress. Further it tested familism as a moderator as endorsing this cultural value may buffer the negative effects of state loneliness. PARTICIPANTS: Participants (n = 220) were Latinx undergraduate students. METHODS: Students reported their loneliness levels and psychological distress twice a day for two weeks using an ecological momentary assessment approach. RESULTS: Results showed that experiencing a higher than usual level of loneliness predicted greater sadness, stress, and anxiety at both the moment-to-moment and day-to-day level. Familism, measured at baseline, only moderated the relationship between loneliness and sadness. CONCLUSIONS: The findings suggest being in a lonely moment may lead to the initiation or amplification of psychological distress immediately and the effects may linger over the day.Supplemental data for this article can be accessed online at https://doi.org/10.1080/07448481.2021.1927051.


Assuntos
Hispânico ou Latino , Solidão , Angústia Psicológica , Estudantes , Humanos , Solidão/psicologia , Estudantes/psicologia , Universidades
12.
J Colloid Interface Sci ; 630(Pt B): 372-381, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36332430

RESUMO

Although widely used as hybrid capacitive deionization (HCDI) electrode material, the low intrinsic conductivity of metal hexacyanometalate (MHCF) severely hinders the fast insertion/extraction of Na+ in/from its 3D framework structure, damaging its desalination performance. Herein, we design a carbon nanotube (CNT) bridged nickel hexacyanoferrate architecture (NiHCF). The highly conductive CNT not only acts as the skeleton for the uniform growth of NiHCF to provide more ion-accessible surface and active sites but also serves as the conductive bridge to connect the NiHCF particles, which prevents the agglomeration of NiHCF particles and facilitates the charge transfer and ion diffusion during the desalination process. Therefore, the HCDI cell assembled by NiHCF/CNT cathode and AC anode exhibits an excellent desalination performance with a high desalination capacity of 29.1 mg g-1 and a superior desalination rate of 7.2 mg g-1 min-1 in 500 mg L-1 NaCl solution. This work provides a facile method for preparing high-performance MHCF-based electrodes for desalination application.

13.
Int J Mol Sci ; 23(22)2022 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-36430648

RESUMO

Drought is a misfortune for agriculture and human beings. The annual crop yield reduction caused by drought exceeds the sum of all pathogens. As one of the gatekeepers of China's "granary", rice is the most important to reveal the key drought tolerance factors in rice. Rice seedlings of Nipponbare (Oryza sativa L. ssp. Japonica) were subjected to simulated drought stress, and their root systems were analyzed for the non-targeted metabolome and strand-specific transcriptome. We found that both DEGs and metabolites were enriched in purine metabolism, and allantoin accumulated significantly in roots under drought stress. However, few studies on drought tolerance of exogenous allantoin in rice have been reported. We aimed to further determine whether allantoin can improve the drought tolerance of rice. Under the treatment of exogenous allantoin at different concentrations, the drought resistant metabolites of plants accumulated significantly, including proline and soluble sugar, and reactive oxygen species (ROS) decreased and reached a significant level in 100 µmol L-1. To this end, a follow-up study was identified in 100 µmol L-1 exogenous allantoin and found that exogenous allantoin improved the drought resistance of rice. At the gene level, under allantoin drought treatment, we found that genes of scavenge reactive oxygen species were significantly expressed, including peroxidase (POD), catalase (CATA), ascorbate peroxidase 8 (APX8) and respiratory burst oxidase homolog protein F (RbohF). This indicates that plants treated by allantoin have better ability to scavenge reactive oxygen species to resist drought. Alternative splicing analysis revealed a total of 427 differentially expressed alternative splicing events across 320 genes. The analysis of splicing factors showed that gene alternative splicing could be divided into many different subgroups and play a regulatory role in many aspects. Through further analysis, we restated the key genes and enzymes in the allantoin synthesis and catabolism pathway, and found that the expression of synthetase and hydrolase showed a downward trend. The pathway of uric acid to allantoin is completed by uric acid oxidase (UOX). To find out the key transcription factors that regulate the expression of this gene, we identified two highly related transcription factors OsERF059 and ONAC007 through correlation analysis. They may be the key for allantoin to enhance the drought resistance of rice.


Assuntos
Alantoína , Oryza , Estresse Fisiológico , Humanos , Alantoína/metabolismo , Alantoína/farmacologia , Seguimentos , Regulação da Expressão Gênica de Plantas , Oryza/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Estresse Fisiológico/genética , Fatores de Transcrição/metabolismo , Transcriptoma , Ácido Úrico/metabolismo , Metaboloma , Fenômenos Fisiológicos Vegetais/genética
14.
ACS Med Chem Lett ; 13(7): 1159-1164, 2022 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-35859885

RESUMO

Activin receptor-like kinase 2 (ALK2) is a transmembrane kinase receptor that mediates the signaling of the members of the TGF-ß superfamily. The aberrant activation of ALK2 has been linked to the rare genetic disorder fibrodysplasia ossificans progressiva (FOP) and diffuse intrinsic pontine glioma (DIPG) that are associated with severely reduced life expectancy in pediatric patients. ALK2 has also been shown to play an essential role in iron metabolism by regulating hepcidin levels and affecting anemia of chronic disease. Thus, selective inhibition of ALK2 has emerged as a promising strategy for the treatment of multiple disorders. Herein, we report the discovery of a novel pyrazolopyrimidines series as highly potent, selective, and orally bioavailable inhibitors of ALK2. Structure-based drug design and systematic structure-activity relationship studies were employed to identify potent inhibitors displaying high selectivity against other ALK subtypes with good pharmacokinetic profiles.

15.
Talanta ; 239: 123086, 2022 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-34871866

RESUMO

Asymptomatic infection of COVID-19 is a global threat for public health. Unfortunately, the study about metabolic dysregulation of asymptomatic infection is barely investigated. Here, we performed carboxylic submetabolome profiling of serum from 62 asymptomatic and 122 control individuals, by a highly sensitive chemical isotope labelling method. Twenty-one discriminative carboxylic features, including 12-hydroxyeicosatetraenoic acid, cholic acid, glycoursodeoxycholic acid and 15,16-dihydroxyoctadeca-9,12-dienoic acid were discovered to be dysregulated in asymptomatic patients. This panel containing 21 carboxylic features could accurately identify asymptomatic patients based on a random forest model, providing an accuracy of 85.7% with only 3.6% false positive rate and 7.1% false negative rate. The dysregulated metabolites found in asymptomatic patients covered several important pathways, such as arachidonic acid metabolism, synthesis of bile acid, ß-oxidation of fatty acids, activation of macrophage and platelet aggregation. This work provided valuable knowledge about serum biomarkers and molecular clues associated with asymptomatic COVID-19 patients.


Assuntos
COVID-19 , Infecções Assintomáticas , Ácidos Graxos , Humanos , Marcação por Isótopo , SARS-CoV-2
16.
Bioorg Med Chem Lett ; 55: 128452, 2022 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-34780900

RESUMO

Activin receptor-like kinase 2 (ALK2) has been implicated as a key target in multiple rare diseases. Herein, we describe the design of a novel bicyclic lactam series of potent and selective ALK2 inhibitors. This manuscript details an improvement in potency of two orders of magnitude from the initial bicyclic structure as well as a two-fold improvement in cellular potency from the original monocyclic inhibitor. Furthermore, we provide a detailed strategy for progressing this project in the future.


Assuntos
Receptores de Ativinas Tipo I/antagonistas & inibidores , Inibidores de Proteínas Quinases/farmacologia , beta-Lactamas/farmacologia , Receptores de Ativinas Tipo I/metabolismo , Relação Dose-Resposta a Droga , Humanos , Estrutura Molecular , Inibidores de Proteínas Quinases/síntese química , Inibidores de Proteínas Quinases/química , Relação Estrutura-Atividade , beta-Lactamas/síntese química , beta-Lactamas/química
17.
Front Plant Sci ; 13: 1043757, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36699859

RESUMO

Oryza Sativa is one of the most important food crops in China, which is easily affected by drought during its growth and development. As a member of the calcium signaling pathway, CBL-interacting protein kinase (CIPK) plays an important role in plant growth and development as well as environmental stress. However, there is no report on the function and mechanism of OsCIPK17 in rice drought resistance. We combined transcriptional and metabonomic analysis to clarify the specific mechanism of OsCIPK17 in response to rice drought tolerance. The results showed that OsCIPK17 improved drought resistance of rice by regulating deep roots under drought stress; Response to drought by regulating the energy metabolism pathway and controlling the accumulation of citric acid in the tricarboxylic acid (TCA) cycle; Our exogenous experiments also proved that OsCIPK17 responds to citric acid, and this process involves the auxin metabolism pathway; Exogenous citric acid can improve the drought resistance of overexpression plants. Our research reveals that OsCIPK17 positively regulates rice drought resistance and participates in the accumulation of citric acid in the TCA cycle, providing new insights for rice drought resistance.

18.
Front Mol Neurosci ; 15: 1076430, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36618828

RESUMO

The retinal degeneration protein RD3 is involved in regulatory processes of photoreceptor cells. Among its main functions is the inhibition of photoreceptor specific membrane guanylate cyclases during trafficking from the inner segment to their final destination in the outer segment. However, any physiological role of RD3 in non-retinal tissue is unsolved at present and specific protein targets outside of retinal tissue have not been identified so far. The family of membrane bound guanylate cyclases share a high homology of their amino acid sequences in their cytoplasmic domains. Therefore, we reasoned that membrane guanylate cyclases that are activated by natriuretic peptides are also regulated by RD3. We analyzed transcript levels of the rd3 gene and natriuretic peptide receptor genes Npr1 and Npr2 in the mouse retina, cerebellum, hippocampus, neocortex, and the olfactory bulb during development from the embryonic to the postnatal stage at P60. The rd3 gene showed a lower expression level than Npr1 and Npr2 (encoding for GC-A and GC-B, respectively) in all tested brain tissues, but was at least one order of magnitude higher in the retina. RD3 and natriuretic peptide receptor GCs co-express in the retina and brain tissue leading to functional tests. We expressed GC-A and GC-B in HEK293T cells and measured the inhibition of GCs by RD3 after activation by natriuretic peptides yielding inhibitory constants around 25 nM. Furthermore, endogenous GCs in astrocytes were inhibited by RD3 to a similar extent. We here show for the first time that RD3 can inhibit two hormone-stimulated GCs, namely GC-A and GC-B indicating a new regulatory feature of these hormone receptors.

19.
Anal Chem ; 93(28): 9904-9911, 2021 07 20.
Artigo em Inglês | MEDLINE | ID: mdl-34227808

RESUMO

Epoxy/dihydroxy-oxylipins are important biologically active compounds that are mainly formed from polyunsaturated fatty acids (PUFAs) in the reactions catalyzed by the cytochrome P450 (CYP 450) enzyme. The analysis of epoxy/dihydroxy-oxylipins would be helpful to gain insights into their landscape in living organisms and provide a reference for the biological studies of these compounds. In this work, we employed chemical labeling-assisted liquid chromatography (LC) coupled with high-resolution mass spectrometry (CL-LC-HRMS) to establish a highly sensitive and specific method for screening and annotating epoxy/dihydroxy-oxylipins in biological samples. The isotope reagents 2-dimethylaminoethylamine (DMED) and DMED-d4 were employed to label epoxy/dihydroxy-oxylipins containing carboxyl groups so as to improve the analysis selectivity and MS detection sensitivity of epoxy/dihydroxy-oxylipins. Based on a pair of diagnostic ions with a mass-to-charge ratio (m/z) difference of 15.995 originating from the fragmentation of derivatives via high-energy collision dissociation (HCD), the potential epoxy/dihydroxy-oxylipins were rapidly screened from the complex matrix. Furthermore, the epoxy/dihydroxy groups could be readily localized by the diagnostic ion pairs, which enabled us to accurately annotate the epoxy/dihydroxy-oxylipins detected in biological samples. The applicability of our method was demonstrated by profiling epoxy/dihydroxy-oxylipins in human serum and heart samples from mice with high-fat diet (HFD). By the proposed method, a total of 32 and 62 potential epoxy/dihydroxy-oxylipins including 42 unreported ones were detected from human serum and the mice heart sample, respectively. Moreover, the relative quantitative results showed that most of the potential epoxy/dihydroxy-oxylipins, especially the oxidation products of linoleic acid (LA) or α-linolenic acid (ALA), were significantly decreased in the heart of mice with HFD. Our developed method is of high specificity and sensitivity and thus is a promising tool for the identification of novel epoxy/dihydroxy-oxylipins in biological samples.


Assuntos
Isótopos , Oxilipinas , Animais , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Sistema Enzimático do Citocromo P-450 , Humanos , Espectrometria de Massas , Camundongos
20.
Front Psychol ; 12: 647544, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34093329

RESUMO

Adolescent mothers experience poorer sleep than adult mothers, and Latina adolescent mothers are at greater risk of postpartum depression compared with other racial/ethnic groups. However, social support may be protective against the negative effects of poor sleep in this population. The current study examined (1) associations between the quality and quantity of Latina adolescent mothers' sleep and mental health (depressive symptoms and anxiety), and (2) whether social support buffered the effects of poor sleep on mental health. A sample of Latina adolescent mothers (N = 84) from an agricultural region in the United States reported on their sleep duration/quality, social support from family, friends, and significant others, and their depressive and anxiety symptoms. Results showed that adolescent mothers reported poorer sleep than pediatric recommendations, and poorer sleep quality was associated with greater depressive and anxiety symptoms. Interestingly, when adolescent mothers reported better sleep, they had fewer depressive symptoms in the context of high support from friends compared with low support from friends. Sleep is important for mental health in Latina adolescent mothers, and better sleep combined with strong social support has positive associations with mental health in this population. Findings hold implications for improving mental health in adolescent mothers.

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