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1.
Bol. latinoam. Caribe plantas med. aromát ; 23(4): 636-644, jul. 2024. graf, tab
Article in English | LILACS | ID: biblio-1538072

ABSTRACT

Thechemical composition, antioxidant and antimicrobial activities of the essential oil from aerial parts (leaves and flowers) of Chuquiraga arcuataHarling grown in the Ecuadorian Andes were studied. One hundred and twenty-six compounds were identified in the essential oil. Monoterpene hydrocarbons (45.8%) and oxygenated monoterpenes (44.1%) had the major percentages. The most abundant compounds were camphor (21.6%), myrcene (19.5%), and 1,8-cineole (13.4%). Antioxidant activity was examined using DPPH, ABTS,and FRAP assays. The essential oil had a moderate scavenging effect and reduction of ferric ion capacity through FRAP assay. Antimicrobial activity of the essential oil was observed against four pathogenic bacteria and a fungus. The essential oil exhibited activity against all microorganism strains under test, particularly against Candida albicansand Staphylococcus aureuswith MICs of 2.43-12.10 µg/mL.


Se estudió la composición química, actividades antioxidantes y antimicrobianas del aceite esencial procedente de las partes aérea (hojas y flores) de Chuquiraga arcuataHarling cultivadas en los Andes ecuatorianos. Se identificaron 126 compuestos en el aceite esencial. Los hidrocarburos monoterpénicos (45,8%) y los monoterpenos oxigenados (44,1%) tuvieron el mayor porcentaje. Los compuestos más abundantes fueron alcanfor (21,6%), mirceno (19,5%) y 1,8-cineol (13,4%). La actividadantioxidante se examinó mediante ensayos DPPH, ABTS y FRAP. El aceite esencial tuvo un efecto eliminador moderado y una reducción de la capacidad de iones férricos mediante el ensayo FRAP. Se observó actividad antimicrobiana del aceite esencial contra cuatro bacterias y un hongo patógenos. El aceite esencial mostró actividad contra todas las cepas de microorganismos bajo prueba, particularmente contra Candida albicansy Staphylococcus aureuscon CMI de 2,43-12,10 µg/mL.


Subject(s)
Oils, Volatile/chemistry , Plant Extracts/chemistry , Asteraceae/chemistry , Anti-Infective Agents/chemistry , Antioxidants/chemistry , Bacteria/drug effects , Oils, Volatile/pharmacology , Plant Extracts/pharmacology , Chromatography, Gas , Plant Leaves/chemistry , Monoterpenes/analysis , Ecuador , Hydrocarbons/analysis , Anti-Infective Agents/pharmacology , Antioxidants/pharmacology
2.
Front Vet Sci ; 10: 1150617, 2023.
Article in English | MEDLINE | ID: mdl-37456968

ABSTRACT

Electroencephalogram (EEG) is a neurophysiological test, which is widely used in human medicine for epilepsy diagnosis and other neurological disorders. For an adequate interpretation, it is necessary to know the electroencephalogram features for different stages of development. Despite the growing interest in its implementation in veterinary medicine, standardized descriptions of the EEG features of the different stages of brain development in dogs are restricted to studies with limited number of dogs and limited age groups. In this research, the electroencephalographic recording of 72 dogs of different breeds and ages was carried out under xylazine sedation to determine tracing characteristics by visual analysis and through statistical analysis of power spectrum. To establish the EEG features of recordings, 3 essential aspects were selected: (a) the presence or absence of slow waves of 4 to 6-7 Hz; (b) the comparison of the electrical activity recorded in the temporal and dorsal cortex channels; and (c) the visual increase of the alpha activity. Visual analysis on both reference and bipolar montage was performed by the authors and additionally blindly corroborated by two human neurophysiologists. The results allowed us to differentiate 5 age groups: 0-5, 6-11, 12-17, 18-23, and >24 months. Statistical analysis of the power spectrum was performed by analysis of variance (ANOVA) with a completely randomized design (CRD) under factorial arrangement by observing the effect of ages, channels and electroencephalographic rhythms on relative power. The results obtained matched those observed in the visual analysis. According to our results, the characteristics of the EEG corresponding to the adult animal begin to appear at 12 months of age but stabilize after 24 months of age. In this case, the evident differences in the processes of development and maturation of the neopallium and the rhinencephalon play a determining role. Our results differ from those obtained by other authors, probably due to the addition of a deep electrode that facilitates the recording of temporal cortical activity and its deeper rhinencephalic connections.

3.
Front Public Health ; 10: 1016353, 2022.
Article in English | MEDLINE | ID: mdl-36276355

ABSTRACT

Objective: This paper identifies varying contemporary and dynamic effects of socio-economic factors on individuals' decisions to allocate their time to physical activities when the intensity of these activities comes into play. Methods: Based on repeated cross-sectional data sourced from the Argentinean National Risk Factor Surveys of 2005, 2009, and 2013, we developed 18 fictitious cohorts to set up a pseudo panel. To address endogeneity problems, four econometric specifications were estimated: OLS, Heckman two-stage model, fixed- and random-effects models. Results: We find that changes in the opportunity cost of time are highly significant and provide shifts in individuals' decisions regarding the allocation of their time to physical activity consumption. When considering the intensity at which physical activities are consumed, increased income impacts less, suggesting that individuals faced with a wage increase reduce the time of consumption but increase its intensity. An interesting finding is that employed people consume more physical activity than inactive individuals. This indicates that the substitution effect produced by an increase in the wage rate is less than the income effect. Additionally, the increase in the coefficient of employed persons is greater when the intensity factor is considered, indicating that for employed individuals a trade-off between time and intensity is generated. We also found that higher levels of education increase the probability of participating in physical activities, but decrease the time spent in such activities. Furthermore, there are heterogeneous impacts on physical activity consumption between males and females, which can be observed in the strong effect of household production for women with at least one child. Finally, such impacts remain in a variety of estimated specifications. Conclusions: These results may be useful in order to suggest some tools for the design of interventions that are aimed at increasing participation in physical activities.


Subject(s)
Exercise , Income , Female , Humans , Male , Cross-Sectional Studies , Economic Factors , Sedentary Behavior
4.
Animals (Basel) ; 12(14)2022 Jul 13.
Article in English | MEDLINE | ID: mdl-35883341

ABSTRACT

The IUCN has listed the long-whiskered owlet (Xenoglaux loweryi) as vulnerable due to the presence of few geographic records, its restricted range, and anthropogenic threats. Its natural history and ecology are largely unknown, and its distribution is widely debated; therefore, there is an urgent need for the real-time conservation of X. loweryi. In this study, 66 geo-referenced records of X. loweryi, 18 environmental variables, and the maximum entropy model (MaxEnt) have been used to predict the current and future (2050 and 2070) potential distribution of X. loweryi in the Amazonas and San Martin regions of northwestern Peru. In fact, under current conditions, areas of "high", "moderate", and "low" potential habitat suitability cover 0.16% (140.85 km2), 0.46% (416.88 km2), and 1.16% (1048.79 km2) of the study area, respectively. Moreover, under future conditions, the "high", "moderate", and "low" probability areas present profits and losses in terms of habitat suitability. Based on the environmental variables, this species mostly inhabits areas with a forest fraction with presence of trees with an emergent tree canopy of ~10-30 metres and depends on Yunga montane forest habitats with high humidity but it is not dependent on bare cover area, crops, or grasslands. Nevertheless, most of the current and future distribution areas are not part of the protected natural areas of Amazonas and San Martin. Additionally, the combination of climate change and anthropogenic activities contribute to further losses of this species habitat. Therefore, from the management point of view, corrective and preventive actions will help to preserve this species over time.

5.
Rev. colomb. quím. (Bogotá) ; 50(3): 16-23, Sep.-Dec. 2021. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1388983

ABSTRACT

Abstract Over time, the effective resistance mechanisms to various first- and second-line drugs against the disease of tuberculosis make its treatment extremely difficult. This work presents a new approach to synthesizing a hybrid of antituberculosis medications: isoniazid (INH) and pyrazinamide (PZA). The synthesis was performed using ultrasound-assisted synthesis to obtain an overall yield of 70%, minimizing the reaction time from 7 to 1 h. The evaluation of the biological activity of the hybrid (compound 2) was tested using the tetrazolium microplate assay (TEMA), showing inhibition in the growth of Mycobacterium tuberculosis H37Rv at a concentration of 0.025 mM at pH 6.0 and 6.7.


Resumen Debido a los grandes mecanismos de resistencia a lo largo del tiempo de diversos fármacos de primera y segunda línea contra la enfermedad de la tuberculosis, el tratamiento sigue dificultándose. Este trabajo presenta un nuevo enfoque para sintetizar un híbrido de fármacos antituberculosos: isoniazida (INH) y pirazinamida (PZA). La síntesis fue asistida por ultrasonido con el fin de obtener un rendimiento global del 70%, minimizando el tiempo de reacción de 7 a ' h. La evaluación de la actividad biológica del híbrido (compuesto 2) se probó usando el ensayo de microplaca de tetrazolio (TEMA), que mostró una inhibición en el crecimiento de Mycobacterium tuberculosis H37Rv a una concentración de 0,025 mM a pH 6,0 y 6,7.


Resumo Devido aos grandes mecanismos de resistência ao longo do tempo a diversos fármacos de primeira e segunda linha contra a tuberculose, o que torna seu tratamento extremamente difícil. Este trabalho apresenta uma nova abordagem para sintetizar um híbrido de fármacos antituberculose: isoniazida (INH) e pirazinamida (PZA) A síntese foi realizada utilizando a síntese assistida por ultrassom de forma a obter um rendimento global de 70%, minimizando o tempo de reação de 7 h para ' h. A avaliação da atividade biológica do híbrido (composto 2) foi testada utilizando o ensaio de microplaca de tetrazólio (TEMA), mostrando uma inibição no crescimento de Mycobacterium tuberculosis H37Rv na concentração de 0,025 mM em pH 6,0 e 6,7.

6.
PLoS Negl Trop Dis ; 15(10): e0009899, 2021 10.
Article in English | MEDLINE | ID: mdl-34705820

ABSTRACT

Poly(A) Binding Proteins (PABPs) are major eukaryotic RNA-binding proteins (RBPs) with multiple roles associated with mRNA stability and translation and characterized mainly from multicellular organisms and yeasts. A variable number of PABP homologues are seen in different organisms however the biological reasons for multiple PABPs are generally not well understood. In the unicellular Leishmania, dependent on post-transcriptional mechanisms for the control of its gene expression, three distinct PABPs are found, with yet undefined functional distinctions. Here, using RNA-immunoprecipitation sequencing analysis we show that the Leishmania PABP1 preferentially associates with mRNAs encoding ribosomal proteins, while PABP2 and PABP3 bind to an overlapping set of mRNAs distinct to those enriched in PABP1. Immunoprecipitation studies combined to mass-spectrometry analysis identified RBPs differentially associated with PABP1 or PABP2, including RBP23 and DRBD2, respectively, that were investigated further. Both RBP23 and DRBD2 bind directly to the three PABPs in vitro, but reciprocal experiments confirmed preferential co-immunoprecipitation of PABP1, as well as the EIF4E4/EIF4G3 based translation initiation complex, with RBP23. Other RBP23 binding partners also imply a direct role in translation. DRBD2, in contrast, co-immunoprecipitated with PABP2, PABP3 and with RBPs unrelated to translation. Over 90% of the RBP23-bound mRNAs code for ribosomal proteins, mainly absent from the transcripts co-precipitated with DRBD2. These experiments suggest a novel and specific route for translation of the ribosomal protein mRNAs, mediated by RBP23, PABP1 and the associated EIF4E4/EIF4G3 complex. They also highlight the unique roles that different PABP homologues may have in eukaryotic cells associated with mRNA translation.


Subject(s)
Leishmania/metabolism , Poly(A)-Binding Proteins/metabolism , Protozoan Proteins/metabolism , RNA, Messenger/metabolism , RNA-Binding Proteins/metabolism , Ribosomal Proteins/metabolism , Leishmania/genetics , Poly(A)-Binding Proteins/genetics , Protein Binding , Protein Biosynthesis , Protozoan Proteins/genetics , RNA, Messenger/genetics , RNA-Binding Proteins/genetics , Ribosomal Proteins/genetics
7.
Metabolomics ; 17(5): 40, 2021 04 16.
Article in English | MEDLINE | ID: mdl-33864540

ABSTRACT

INTRODUCTION: Brain death (BD) is the irreversible cessation of all functions of the entire brain, including the brainstem. Cerebrospinal fluid (CSF) is a biological liquid that circulates in brain and spine. Metabolomics is able to reveal the response of biological systems to diverse factors in a specific moment or condition. Therefore, the study of this neurological condition through metabolic profiling using high resolution Nuclear Magnetic Resonance (NMR) spectroscopy is important for understanding biochemical events. OBJECTIVES: The aim of the current study is to identify the metabolomics signature of BD using 1H-NMR spectroscopy in human CSF. METHODS: 1H-NMR spectroscopy has been employed for metabolomic untargeted analysis in 46 CSF samples: 22 control and 24 with BD. Spectral data were further subjected to multivariate analysis. RESULTS: Statistically significant multivariate models separated subject's samples with BD from controls and revealed twenty one discriminatory metabolites. The statistical analysis of control and BD subjects using Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) model resulted in R2X of 0.733 and Q2 of 0.635. An elevation in the concentration of statistically discriminant metabolites in BD was observed. CONCLUSION: This study identifies a metabolic signature associated with BD and the most relevant enriched selected metabolic pathways.


Subject(s)
Brain Death , Metabolomics , Humans , Magnetic Resonance Imaging , Magnetic Resonance Spectroscopy , Metabolic Networks and Pathways
8.
J Fish Biol ; 98(2): 577-582, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33090509

ABSTRACT

We report 24 new records of the Brazilian cownose ray Rhinoptera brasiliensis outside its accepted geographic range. Sequencing of a 442-base pair portion of the mitochondrial NADH dehydrogenase subunit 2 gene for 282 Rhinoptera samples revealed eight records off the east coast of the USA and 16 from the eastern Gulf of Mexico. Both sexes of all life stages were documented in all seasons over multiple years in the Indian River and Lake Worth lagoons, Florida, indicating that their range extends further in the western North Atlantic than previously described.


Subject(s)
Animal Distribution , NADH Dehydrogenase/genetics , Skates, Fish/genetics , Animals , Atlantic Ocean , Female , Florida , Gulf of Mexico , Male , Rivers , Skates, Fish/classification
10.
Front Immunol ; 11: 569358, 2020.
Article in English | MEDLINE | ID: mdl-33117357

ABSTRACT

Atherosclerotic cardiovascular disease is part of chronic immunometabolic disorders such as type 2 diabetes and nonalcoholic fatty liver disease. Their common risk factors comprise hypertension, insulin resistance, visceral obesity, and dyslipidemias, such as hypercholesterolemia and hypertriglyceridemia, which are part of the metabolic syndrome. Immunometabolic diseases include chronic pathologies that are affected by both metabolic and inflammatory triggers and mediators. Important and challenging questions in this context are to reveal how metabolic triggers and their downstream signaling affect inflammatory processes and vice-versa. Along these lines, specific nuclear receptors sense changes in lipid metabolism and in turn induce downstream inflammatory and metabolic processes. The transcriptional activity of these nuclear receptors is regulated by the nuclear receptor corepressors (NCORs), including NCOR1. In this review we describe the function of NCOR1 as a central immunometabolic regulator and focus on its role in atherosclerosis and associated immunometabolic diseases.


Subject(s)
Atherosclerosis/etiology , Atherosclerosis/metabolism , Disease Susceptibility , Metabolic Diseases/etiology , Metabolic Diseases/metabolism , Nuclear Receptor Co-Repressor 1/metabolism , Animals , Atherosclerosis/pathology , Carrier Proteins , Disease Susceptibility/immunology , Energy Metabolism/immunology , Humans , Immunomodulation , Lipid Metabolism , Macrophages/immunology , Macrophages/metabolism , Nuclear Receptor Co-Repressor 1/genetics , Protein Binding , Signal Transduction
11.
Zootaxa ; 4803(2): zootaxa.4803.2.3, 2020 Jun 26.
Article in English | MEDLINE | ID: mdl-33056018

ABSTRACT

Herein we consolidate the information available concerning the biodiversity of batoid fishes in the northern Gulf of Mexico, including nearly 70 years of survey data collected by the National Marine Fisheries Service, Mississippi Laboratories and their predecessors. We document 41 species proposed to occur in the northern Gulf of Mexico. However, the validity of several of these reports and their associated data is questioned. In addition, we provide information and remarks concerning the distribution, conservation status, taxonomy and recorded history for each species covered.


Subject(s)
Elasmobranchii , Fishes , Animals , Biodiversity , Gulf of Mexico
13.
J Adv Vet Anim Res ; 7(2): 320-323, 2020 Jun.
Article in English | MEDLINE | ID: mdl-32607364

ABSTRACT

OBJECTIVE: Acute hepatopancreatic necrosis disease is an emerging infectious disease of Penaeus species. The causative agent is Vibrio species, which dispels binary toxin similar to pirA and pirB, which causes mortality in infected shrimp. The aim of this research was to investigate the evolutionary relationship of pirA and pirB homologous genes present in this Asia and Costa Rica in silico. MATERIALS AND METHODS: The sequences for in silico analysis were all retrieved from the Basic Local Alignment Search Tool Nucleotide (BLASTN) tool of the National Center for Biotechnology Center. For pirA, a total of 25 sequences submitted from different Asian countries and Costa Rica were retrieved for analysis. Meanwhile, for pirB, a total of 11 sequences submitted from five Asian countries were retrieved. Sequences were aligned using the CLUSTAL W alignment tool under Molecular Evolutionary Genetics Analysis (MEGA) 7 software. The evolutionary history was then estimated using the Unweighted Pair Group Method with Arithmetic mean (UPGMA) method, whereas the evolutionary distances were determined using the maximum composite likelihood model with 1,000 bootstrap replications. RESULTS AND DISCUSSION: The results show that, among 27 DNA sequences analyzed for pirA gene, three groups were generated, while for pirB, 13 DNA sequences yielded only one group. The analysis revealed low genetic variation among isolates for both pirA and pirB genes. CONCLUSION: This result suggests that the low frequency of polymorphism and geographic location cannot be attributed to the differences in V. parahaemolyticus isolates in Asian countries relative to Costa Rican isolates in pirA and pirB genes.

14.
J Pediatr ; 222: 213-220.e5, 2020 07.
Article in English | MEDLINE | ID: mdl-32586526

ABSTRACT

OBJECTIVE: To test whether variants in ADRB1 and CYP2C9 genes identify subgroups of individuals with differential response to treatment for Marfan syndrome through analysis of data from a large, randomized trial. STUDY DESIGN: In a subset of 250 white, non-Hispanic participants with Marfan syndrome in a prior randomized trial of atenolol vs losartan, the common variants rs1801252 and rs1801253 in ADRB1 and rs1799853 and rs1057910 in CYP2C9 were analyzed. The primary outcome was baseline-adjusted annual rate of change in the maximum aortic root diameter z-score over 3 years, assessed using mixed effects models. RESULTS: Among 122 atenolol-assigned participants, the 70 with rs1801253 CC genotype had greater rate of improvement in aortic root z-score compared with 52 participants with CG or GG genotypes (Time × Genotype interaction P = .005, mean annual z-score change ± SE -0.20 ± 0.03 vs -0.09 ± 0.03). Among participants with the CC genotype in both treatment arms, those assigned to atenolol had greater rate of improvement compared with the 71 of the 121 assigned to losartan (interaction P = .002; -0.20 ± 0.02 vs -0.07 ± 0.02; P < .001). There were no differences in atenolol response by rs1801252 genotype or in losartan response by CYP2C9 metabolizer status. CONCLUSIONS: In this exploratory study, ADRB1-rs1801253 was associated with atenolol response in children and young adults with Marfan syndrome. If these findings are confirmed in future studies, ADRB1 genotyping has the potential to guide therapy by identifying those who are likely to have greater therapeutic response to atenolol than losartan.


Subject(s)
Atenolol/therapeutic use , Cytochrome P-450 CYP2C9/genetics , Gene Expression Regulation , Losartan/therapeutic use , Marfan Syndrome/drug therapy , Receptors, Adrenergic, beta-1/genetics , Adolescent , Adrenergic beta-1 Receptor Antagonists/therapeutic use , Adult , Angiotensin II Type 1 Receptor Blockers/therapeutic use , Child , Child, Preschool , Cytochrome P-450 CYP2C9/biosynthesis , DNA/genetics , Female , Follow-Up Studies , Genotype , Humans , Infant , Male , Marfan Syndrome/genetics , Marfan Syndrome/metabolism , Receptors, Adrenergic, beta-1/biosynthesis , Retrospective Studies , Young Adult
15.
J Clin Microbiol ; 58(6)2020 05 26.
Article in English | MEDLINE | ID: mdl-32238433

ABSTRACT

The prevalence of carbapenem-resistant Pseudomonas aeruginosa is increasing. Identification of carbapenemase-producing P. aeruginosa will have therapeutic, epidemiological, and infection control implications. This study evaluated the performance of the EDTA-modified carbapenem inactivation method (eCIM) in tandem with the modified carbapenem inactivation method (mCIM) against a large collection of clinical P. aeruginosa isolates (n = 103) to provide clinicians a phenotypic test that not only identifies carbapenemase production but also distinguishes between metallo-ß-lactamase and serine-carbapenemase production in P. aeruginosa The mCIM test was performed according to Clinical and Laboratory Standards Institute guidelines, while the eCIM was conducted as previously described for Enterobacteriaceae Test performance was compared to the genotypic profile as the reference. mCIM testing successfully categorized 91% (112/123) of P. aeruginosa isolates as carbapenemases or non-carbapenemase producers, with discordant isolates being primarily Guiana extended-spectrum (GES)-type producers. To increase the sensitivity of the mCIM for GES-harboring isolates, a double inoculum, prolonged incubation, or both was evaluated, with each modification improving sensitivity to 100% (12/12). Upon eCIM testing, all Verona integrin-encoded metallo-ß-lactamases (VIM; n = 27) and New Delhi metallo-ß-lactamases (NDM; n = 13) tested had 100% concordance to their genotypic profiles, whereas all Klebsiella pneumoniae carbapenemase (KPC; n = 8) and GES (n = 12) isolates tested negative, as expected, in the presence of EDTA. The eCIM failed to identify all imipenemase (IMP)-producing (n = 22) and Sao Paulo metallo-ß-lactamase (SPM)-producing (n = 14) isolates. KPC-, VIM-, and NDM-producing P. aeruginosa were well defined by the conventional mCIM and eCIM testing methods; additional modifications appear required to differentiate GES-, IMP-, and SPM-producing isolates.


Subject(s)
Pseudomonas aeruginosa , beta-Lactamases , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Brazil , Carbapenems/pharmacology , Edetic Acid/pharmacology , Humans , Microbial Sensitivity Tests , Pseudomonas aeruginosa/genetics , beta-Lactamases/genetics
16.
J Anim Sci ; 98(4)2020 Apr 01.
Article in English | MEDLINE | ID: mdl-32211788

ABSTRACT

Mammals have a circadian rhythm that is synchronized by a master clock located in the hypothalamic suprachiasmatic nucleus (SCN). The SCN regulates additional clocks located in peripheral tissues, including some involved in endocrine or reproductive functions. Studies in humans and mice report that molecular clocks also exist in the placenta. However, little is known about the presence of "Clock genes," namely Circadian Locomotor Output Cycles Kaput (CLOCK), Brain and Muscle Arnt-Like 1 (BMAL1), Period 1 (PER1), Period 2 (PER2), Cryptochrome 1 (CRY1), and Cryptochrome 2 (CRY2), in equine placenta. Pregnancy length in mares varies and shows fluctuations in hormone concentrations throughout pregnancy. We postulate that similar to humans and mice, Clock genes are present in the horse placentas. Our goal was to determine if relative levels of clock genes were different between placentas associated with males and female fetuses or correlated with gestational length. We used polymerase chain reaction and immunofluorescence to study the presence of CLOCK, BMAL1, PER1, PER2, CRY1, and CRY2 in full-term mare placentas. Clock genes were present in all placentas, with significant lower levels of CRY2 and CLOCK in placentas that were associated with male fetuses. There was no association between relative levels of Clock genes and gestational length. These data provide the stage for future studies aimed at uncovering a function for Clock genes in the horse placenta.


Subject(s)
CLOCK Proteins/metabolism , Horses/physiology , Placenta/metabolism , Animals , CLOCK Proteins/genetics , Female , Gene Expression Regulation , Male , Pregnancy
17.
Mol Cell Endocrinol ; 505: 110719, 2020 04 05.
Article in English | MEDLINE | ID: mdl-31972331

ABSTRACT

Thyroglobulin (TG), a large glycosylated protein secreted by thyrocytes into the thyroid follicular lumen, plays an essential role in thyroid hormone biosynthesis. Rattus norvegicus TG (rTG) is encoded by a large single copy gene, 186-kb long, located on chromosome 7 composed of 48 exons encoding a 8461-kb mRNA. Although the TG gene displays sequence variability, many missense mutations do not impose any adverse effect on the TG protein, whereas other nucleotide substitutions may affect its TG stability and/or TG intracellular trafficking. In order to gain a further understanding of the protein domains regulating its intracellular fate, we cloned a full-length cDNA from rTG into the pcDNA6/V5-His B expression vector. However, transient expression of the cDNA in HEK293T cells showed that the encoded protein was not a wild-type molecule, as it was unable to be secreted in the culture supernatant. Sequencing analyses revealed three random mutations, which accidentally emerged during the course of cloning: c.1712T>C [p.L571P] in the linker domain (amino acid positions 360 to 604), c.2027A>G [p.Q676R] in TG type 1-6 repeat and c.2720A>G [p.Q907R] in the TG type 1-7 repeat. Expression of cDNAs encoding a combination of two mutations [p.Q676R-p.Q907R], [p.L571P-p.Q907R] or [p.L571P-p.Q676R] indicated that any TG bearing the p.L571P substitution was trapped intracellularly. Indeed, we expressed the single point mutant p.L571P and confirmed that this point mutation was sufficient to cause intracellular retention of mutant TG in HEK293T cells. Endo H analysis showed that the p.L571P mutant is completely sensitive to the enzyme, whereas the will-type TG acquires full N-glycan modifications in Golgi apparatus. This data suggest that the p.L571P mutant contains the mannose-type N-glycan, that was added at the first stage of glycosylation. Complex-type N-glycan formation in the Golgi apparatus does not occur, consistent with defective endoplasmic reticulum exit of the mutant TG. Moreover, predictive analysis of the 3D linker domain showed that the p.L571P mutation would result in a significant protein conformational change. In conclusion, our studies identified a novel amino acid residue within the linker domain of TG associated with its conformational maturation and intracellular trafficking.


Subject(s)
Intracellular Space/metabolism , Mutation/genetics , Thyroglobulin/chemistry , Thyroglobulin/genetics , Amino Acid Sequence , Animals , Base Sequence , DNA, Complementary/genetics , Glycoside Hydrolases/metabolism , HEK293 Cells , Humans , Male , Mutagenesis/genetics , Mutant Proteins/chemistry , Mutant Proteins/metabolism , Protein Domains , Protein Multimerization , Protein Structure, Secondary , Rats, Wistar
18.
Sci Rep ; 9(1): 15934, 2019 11 04.
Article in English | MEDLINE | ID: mdl-31685909

ABSTRACT

Intertidal communities' composition and diversity usually exhibit strong changes in relation to environmental gradients at different biogeographical scales. This study represents the first comprehensive diversity and composition description of polyplacophoran assemblages along the Peruvian Province (SE Pacific, 12°S-39°S), as a model system for ecological latitudinal gradients. A total of 4,775 chitons from 21 species were collected on twelve localities along the Peruvian Province. This sampling allowed us to quantitatively estimate the relative abundance of the species in this assemblage, and to test whether chitons conform to elementary predictions of major biogeographic patterns such as a latitudinal diversity gradient. We found that the species composition supported the division of the province into three ecoregional faunal groups (i.e. Humboldtian, Central Chile, and Araucanian). Though chiton diversity did not follow a clear latitudinal gradient, changes in species composition were dominated by smaller scale variability in salinity and temperature. Body size significantly differed by ecoregions and species, indicating latitudinal size-structure assamblages. In some localities body size ratios differed from a random assemblage, evidencing competition at local scale. Changes in composition between ecoregions influence body size structure, and their overlapping produce vertical size segregation, suggesting that competition coupled with environmental conditions structure these assemblages.

19.
Front Physiol ; 10: 1238, 2019.
Article in English | MEDLINE | ID: mdl-31649551

ABSTRACT

Competition between same-sex organisms, or intra-sexual selection, can occur before and after mating, and include processes such as sperm competition and cryptic female choice. One of the consequences of intra-sexual selection is that male reproductive traits tend to evolve and diverge at high rates. In benthic octopuses, females often mate with more than one male in a single reproductive event, opening the venue for intra-sexual selection at multiple levels. For instance, males transfer spermatophores through hectocotylus, and can remove the spermatophores left by other males. Considering the limited evidence on post-copula competition in benthic octopuses, and the potential to affect the evolution of reproductive traits within octopodids, we put this hypothesis to a test employing a phylogenetic comparative approach. We combined data on hectocotylized arm length (HAL), ligula length (LL), spermatophore length (SL) with a Bayesian molecular phylogeny of 87 species, to analyze how reproductive traits have diverged across lineages and covary with body size (mantle length; ML). First, additionally to ML, we estimated the phylogenetic signal (λ) and mode of evolution (κ) in each reproductive trait. Second, we performed phylogenetic regressions to quantify the association among reproductive traits and their co-variation with ML. This analysis allowed us to estimate the phenotypic change along a branch into the phylogeny, and whether selection may have played a role in the evolution and diversification of specific clades. Estimations of λ were always high (>0.75), indicating concordance between the traits and the topology of the phylogenetic tree. Low values of κ (<1.0) suggested that evolution depends on branch lengths. All reproductive traits exhibiting a positive relation with ML (ß > 0.5 in all cases). Overall, evolutionary rate models applied to the SL-ML regression suggested that octopuses of the family Megaleledonidae have evolved larger spermatophores than expected for their size. The regression HAL-ML indicated that HAL was more variable in Megaleledonidae than in the remaining clades, suggesting that the high divergence across species within this group might partially reflect intra-sexual selection. These results support the hypothesis that, at least in some lineages, sexual selection may account for the divergence in reproductive traits of male octopuses.

20.
Acta bioquím. clín. latinoam ; Acta bioquím. clín. latinoam;53(2): 255-266, jun. 2019. graf, tab
Article in Spanish | LILACS | ID: biblio-1019259

ABSTRACT

Antecedentes. HDL es cuantitativamente la lipoproteína más importante en la mayoría de las especies y la evidencia mecanicista sugiere que HDL tendría un papel en la función inmunológica normal. Probamos la hipótesis que sugiere que las concentraciones plasmáticas de HDL están asociadas con el riesgo de enfermedades autoinmunes. Métodos. Se incluyeron 107.954 y 9.387 individuos con mediciones basales de colesterol-HDL provenientes de 2 estudios de la población general: el Estudio de la Población General de Copenhague y el Estudio del Corazón de Copenhague. Los pacientes fueron seguidos mediante el Registro Nacional Danés de pacientes desde el inicio del período 2003-2015 o 1991-1994 hasta 2017, tiempo durante el cual 4.078 y 1.101 individuos desarrollaron enfermedad autoinmune en los 2 estudios respectivamente. Resultados. En el Estudio de la Población General de Copenhague, en comparación a los individuos con colesterol de HDL =2,0 mmol/L (77 mg/dL), los índices de riesgo para cualquier enfermedad autoinmune, ajustados de manera multifactorial fueron 1,06 (IC 95%, 0,94-1,19) para individuos con colesterol-HDL entre 1,5 y 1,99 mmol/L (58 a 77 mg/dL), 1,18 (IC 95%, 1,04-1,35) para individuos con colesterol-HDL entre 1,0 y 1,49 mmol/L (39 a 58 mg/dL) y 1,84 (IC 95%, 1,52- 2,22) para individuos con colesterol-HDL <1,0 mmol/L (39 mg/dL) (p<0,001 para tendencia). Estos resultados fueron similares cuando: se excluyeron los eventos dentro de los 5 años del inicio del estudio, tanto en mujeres como hombres por separado, eventos en el inicio del estudio, independientemente de la inflamación de bajo grado o concentraciones de triglicéridos, para diferentes niveles de apolipoproteína A1 y para definiciones de punto final más restrictivas. Finalmente, el Estudio del Corazón de Copenhague proporcionó una confirmación independiente. Conclusiones: Los bajos niveles de colesterol-HDL se asocian con un alto riesgo de enfermedad autoinmune en individuos de la población general. Nuestros hallazgos observacionales no pueden determinar la causalidad.


Background. HDL is quantitatively the most important lipoprotein in most species and mechanistic evidence points toward a role for HDL in normal immune function. We tested the hypothesis that concentrations of HDL cholesterol are associated with risk of autoimmune disease. Methods. From 2 studies of the general population-the Copenhagen General Population Study and the Copenhagen City Heart study-we included 107,954 and 9,387 individuals with baseline measurements of HDL cholesterol. These were followed with the national Danish Patient Registry from baseline in 2003-2015 or 1991-1994 through 2017, during which time 4078 and 1101 individuals developed autoimmune disease in the 2 studies. Results. In the Copenhagen General Population Study, compared to individuals with HDL cholesterol =2.0 mmol/L (77 mg/dL), the multifactorially adjusted hazard ratios for any autoimmune disease were 1.06 (95% CI, 0.94-1.19) for individuals with HDL cholesterol of 1.5-1.99 mmol/L (58-77 mg/dL), 1.18 (95% CI, 1.04-1.35) for individuals with HDL cholesterol of 1.0-1.49 mmol/L (39-58 mg/dL), and 1.84 (95% CI, 1.52-2.22) for individuals with HDL cholesterol <1.0 mmol/L (39 mg/dL) (p for trend <0.001). These results were similar when excluding events within 5 years of baseline, in women and men separately, for events at baseline, irrespective of low-grade inflammation or triglyceride concentrations, for the apolipoprotein A1 part of HDL, and for more restrictive end point definitions. Finally, the Copenhagen City Heart Study provided independent confirmation. Conclusions. Low HDL cholesterol level is associated with high risk of autoimmune disease in individuals from the general population. Our observational findings cannot determine causality.


Subject(s)
Humans , Male , Female , Middle Aged , Aged , Autoimmune Diseases/diagnosis , Cholesterol, HDL/blood , Autoimmune Diseases/blood , Epidemiologic Studies , Denmark , Cholesterol, HDL/urine
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