RESUMO
BACKGROUND: Alzheimer's disease (AD) is a neurological condition that may lead to dementia as well as a slow and steady decline in cognitive ability. Finding early signs that may be used in the diagnosis of AD is still a difficult aim to achieve in the field of medical practice. METHODS AND RESULTS: The purpose of this research was to investigate to determine any differences in the gene expression patterns of crystallin mu (CRYM) and sialic acid-binding immunoglobulin-like lectin 10 (SIGLEC10) in whole blood samples obtained from fifty individuals who were diagnosed with AD and fifty individuals as a control group. When compared with controls, it was discovered that the expression of the CRYM gene was substantially decreased in AD patients, but the expression of the SIGLEC10 gene was significantly higher. A positive correlation between CRYM and SIGLEC10 was noticed solely in patients with AD. Furthermore, assessing the diagnostic value of these genes, CRYM and SIGLEC10 transcript levels displayed an area under the curve (AUC) of 0.74 and 0.81, respectively. CONCLUSIONS: These results suggest that alterations in CRYM and SIGLEC10 expression may be implicated in AD pathology and that these genes expression levels can potentially serve as biomarkers for early detection and diagnosis of AD. Nevertheless, further validation of these findings requires the inclusion of more extensive and heterogeneous cohorts. The findings derived from this study possess the capability to offer a significant contribution towards the progression of innovative diagnostic and therapeutic strategies for AD.
Assuntos
Doença de Alzheimer , Cristalinas mu , Humanos , Doença de Alzheimer/diagnóstico , Doença de Alzheimer/genética , Biomarcadores , Expressão Gênica , Lectinas/genética , Receptores de Superfície CelularRESUMO
BACKGROUND: Alzheimer's disease (AD) is a multifaceted neurological ailment affecting more than 50 million individuals globally, distinguished by a deterioration in memory and cognitive abilities. Investigating neurotrophin growth factors could offer significant contributions to understanding AD progression and prospective therapeutic interventions. METHODS AND RESULTS: The present investigation collected blood samples from 50 patients diagnosed with AD and 50 healthy individuals serving as controls. The mRNA expression levels of neurotrophin growth factors and their receptors were measured using quantitative PCR. A Bayesian regression model was used in the research to assess the relationship between gene expression levels and demographic characteristics such as age and gender. The correlations between variables were analyzed using Spearman correlation coefficients, and the diagnostic potential was assessed using a Receiver Operating Characteristic curve. NTRK2, TrkA, TrkC, and BDNF expression levels were found to be considerably lower (p-value < 0.05) in the blood samples of AD patients compared to the control group. The expression of BDNF exhibited the most substantial decrease in comparison to other neurotrophin growth factors. Correlation analysis indicates a statistically significant positive association between the genes. The ROC analysis showed that BDNF exhibited the greatest Area Under the Curve (AUC) value of 0.76, accompanied by a sensitivity of 70% and specificity of 66%. TrkC, TrkA, and NTRK2 demonstrated considerable diagnostic potential in distinguishing between cases and controls. CONCLUSION: The observed decrease in the expression levels of NTRK2, TrkA, TrkC, and BDNF in AD patients, along with the identified associations between specific genes and their diagnostic capacity, indicate that these expressions have the potential to function as biomarkers for the diagnosis and treatment of AD.
Assuntos
Doença de Alzheimer , Humanos , Doença de Alzheimer/diagnóstico , Doença de Alzheimer/genética , Teorema de Bayes , Fator Neurotrófico Derivado do Encéfalo/genética , Receptores Proteína Tirosina Quinases , BiomarcadoresRESUMO
Charcot-Marie-Tooth (CMT) comprises a group of hereditary neuropathies with clinical, epidemiological, and molecular heterogeneity in which variants in more than 80 different genes have been reported. One of the important genes which cause 5% of all CMT cases is Myelin protein zero (P0, MPZ). Variants in this gene have been reported in association with different forms of CMT including classical CMT1, severe DSS (CMT3B), DI-CMT, CMT2I and CMT2J with autosomal dominant (AD) inheritance. To our knowledge, MPZ variants have not been described in autosomal recessive (AR) form of CMT in previous studies. Moreover, its complete deletion has not been reported in human. Here, we described clinical characteristics of a patient with CMT symptoms who demonstrated manifestations of the disease late in his life. We performed exome sequencing for identifying CMT subtype and its associated gene, and follow that co-segregation analysis has been done to characterize inheritance pattern of the disorder. Through using exome sequencing, we identified a novel 4074 bp homozygote deletion which encompasses all 6 exons of the MPZ gene in this patient. After identifying the alteration, variant confirmation and co-segregation analysis have been performed by using specific primers. Our result revealed that the patient's parents were heterozygous for the alteration and they did not show any symptoms of CMT. Although most MPZ variants have been described with early onset CMT with AD pattern of inheritance, the reported patient in our study had late onset form and his parents did not show any symptoms. Considering substantial role of MPZ protein in the biogenesis of peripheral nervous system (PNS) myelin, we proposed that there should be another protein in PNS that compensates for lack of MPZ protein. Taken together, our finding is the first report of MPZ association with AR form of CMT with late onset features. Moreover, our results propose the presence of another protein in PNS myelin biogenesis and its assembly. However, functional studies alongside with other molecular studies are needed to confirm our results and identify the proposed protein.
Assuntos
Doença de Charcot-Marie-Tooth , Proteína P0 da Mielina , Humanos , Doença de Charcot-Marie-Tooth/genética , Doença de Charcot-Marie-Tooth/diagnóstico , Éxons , Mutação/genética , Proteína P0 da Mielina/genética , Bainha de Mielina , Proteínas/genéticaRESUMO
Alzheimer's disease (AD) is a global health problem due to its complexity, which frequently makes the development of treatment methods extremely difficult. Therefore, new methodologies are necessary to investigate the pathophysiology of AD and to treat AD. The interaction of immune modulation and neurodegeneration has added new dimensions in current knowledge of AD etiology and offers an attractive opportunity for the discovery of novel biomarkers and therapies. Using quantitative polymerase chain reaction, we compared the expression levels of inhibitory B7 family members (B7-1, B7-2, B7-H1, B7-DC, B7-H3, B7-H4, B7-H5, B7-H7, and ILDR2), as immune regulators, in the peripheral blood of late-onset AD (LOAD) patients (n = 50) and healthy individuals (n = 50). The levels of B7-2, B7-H4, ILDR2, and B7-DC expression were significantly higher in-patient blood samples than in control blood samples. Furthermore, we discovered a substantial positive correlation between all gene expression levels. In addition, the current study indicated that ILDR2, B7-H4, B7-2, and B7-DC might serve as diagnostic biomarkers to identify LOAD patients from healthy persons. The present work provides additional evidence for the significance of inhibitory B7 family members to the etiology of LOAD.
Assuntos
Doença de Alzheimer , Inibidor 1 da Ativação de Células T com Domínio V-Set , Humanos , Inibidor 1 da Ativação de Células T com Domínio V-Set/genética , Inibidor 1 da Ativação de Células T com Domínio V-Set/metabolismo , Doença de Alzheimer/genética , BiomarcadoresRESUMO
Schizophrenia (SCZ) is a severe mental disorder with an unknown pathophysiology. Brain-Derived Neurotrophic Factor (BDNF) is a neurotrophin that has been associated with synapse plasticity, learning, and memory, as well as neurodevelopment and neuroprotection. The importance of neurodevelopmental and neurotoxicity-related components in the pathophysiology of SCZ has been highlighted in research on the neurobiology of this disease. The purpose of this research is to investigate the significant expression of two variables, tristetraprolin (TTP) and miR-16, which are known to be regulators of BDNF expression. Fifty Iranian Azeri SCZ patients were enrolled, and fifty healthy volunteers were age- and gender-matched as controls. A quantitative polymerase chain reaction measured the expression levels of the TTP and miR-16 in the peripheral blood (PB) of SCZ patients and healthy people. TTP expression levels in patients were higher than in controls, regardless of gender or age (posterior beta = 1.532, adjusted P-value = 0.012). TTP and miR-16 expression levels were found to be significantly correlated in both SCZ patients and healthy controls (r = 0.701, P < 0.001 and r = 0.777, P < 0.001, respectively). Due to the increased expression of TTP in SCZ and the existence of a significant correlation between TTP and miR-16, which helps to act on target mRNAs with AU-rich elements, this mechanism can be considered an influencing factor in SCZ.
Assuntos
MicroRNAs , Esquizofrenia , Humanos , Fator Neurotrófico Derivado do Encéfalo/genética , Esquizofrenia/genética , Irã (Geográfico) , MicroRNAs/genéticaRESUMO
Schizophrenia (SCZ) is known as a complicated mental disease with an unknown etiology. The microdeletion of 22q11.2 is the most potent genetic risk factor. Researchers are still trying to find which genes in the deletion region are linked to SCZ. MIR185, encoding microRNA (miR)-185, is present in the minimal 1.5 megabase deletion. Nonetheless, the miR-185 expression profile and its corresponding target genes in animal models and patients with 22q11.2 deletion syndrome (22q11.2DS) imply that more study is required about miR-185 and its corresponding downstream pathways within idiopathic SCZ. The expression of hsa-miR-185-5p and its corresponding target gene, shisa family member 7 (SHISA7), sometimes called CKAMP59, were evaluated in the peripheral blood (PB) samples of Iranian Azeri patients with idiopathic SCZ and healthy subjects, matched by gender and age as control groups by quantitative polymerase chain reaction (qPCR). Fifty SCZ patients (male/female: 22/28, age (mean ± standard deviation (SD)): 35.9 ± 5.6) and 50 matched healthy controls (male/female: 23/27, age (mean ± SD): 34.7 ± 5.4) were enrolled. The expression of hsa-miR-185-5p in the PB samples from subjects with idiopathic SCZ was substantially lower than in that of control groups (posterior beta = -0.985, adjusted P-value < 0.0001). There was also a difference within the expression profile between female and male subgroups (posterior beta = -0.86, adjusted P-value = 0.046 and posterior beta = -1.015, adjusted P-value = 0.004, in turn). Nevertheless, no significant difference was present in the expression level of CKAMP59 between PB samples from patients and control groups (adjusted P-value > 0.999). The analysis of the receiver operating characteristic (ROC) curve suggested that hsa-miR-185-5p may correctly distinguish subjects with idiopathic SCZ from healthy people (the area under curve (AUC) value: 0.722). Furthermore, there was a strong positive correlation between the expression pattern of the abovementioned genes in patients with SCZ and healthy subjects (r = 0.870, P < 0.001 and r = 0.812, P < 0.001, respectively), indicating that this miR works as an enhancer. More research is needed to determine if the hsa-miR-185-5p has an enhancer activity. In summary, this is the first research to highlight the expression of the miR-185 and CKAMP59 genes in the PB from subjects with idiopathic SCZ. Our findings suggest that gene expression alterations mediated by miR-185 may play a role in the pathogenesis of idiopathic and 22q11.2DS SCZ. It is worth noting that, despite a substantial and clear relationship between CKAMP59 and hsa-miR-185-5p, indicating an interactive network, their involvement in the development of SCZ should be reconsidered based on the whole blood sample since the changed expression level of CKAMP59 was not significant. Further research with greater sample sizes and particular leukocyte subsets can greatly make these results stronger.
Assuntos
Síndrome de DiGeorge , MicroRNAs , Esquizofrenia , Adulto , Animais , Síndrome de DiGeorge/genética , Regulação para Baixo , Feminino , Humanos , Irã (Geográfico) , Masculino , MicroRNAs/metabolismo , Esquizofrenia/genéticaRESUMO
AIM AND OBJECTIVE: This study aims to assess the effects of aloe vera toothpaste on dental plaques and gingivitis. MATERIALS AND METHODS: This single-center, single-blind, randomized, two-period crossover study was performed on 20 dental students with a mean age of 24.5 ± 4 years with gingivitis. Subjects were randomly assigned to two groups (n = 10). After 14 days of trial period, plaque index (PI) and gingival index (GI) were assessed for each group. The first group used aloe vera toothpaste for 30 days and then their PI and GI were recorded. A 2-week washout period was allowed and then the subjects used fluoride toothpaste for the next 30 days and underwent PI and GI assessment again. This order was reversed in group 2. RESULTS: Toothpaste-containing aloe vera showed no significant improvement in the GI and PI scores as compared with a fluoride-containing dentifrice. PI was 2.14 ± 1.3 at baseline and 1.84 ± 1.02 in 30 days (p <0.098). GI was 0.62 ± 0.74 at baseline and 0.25 ± 0.46 at 30 days (p <0.068). During the trial, no side effects were seen due to the use of aloe vera or fluoride toothpaste. CONCLUSION: The effect of aloe vera toothpaste on PI and GI was similar to that of fluoride toothpaste and it seems that this toothpaste can be used as an alternative to a chemical toothpaste. CLINICAL SIGNIFICANCE: The use of an aloe vera toothpaste in improving the progression of gingivitis can be evaluated.
Assuntos
Aloe , Placa Dentária , Gengivite , Adulto , Estudos Cross-Over , Índice de Placa Dentária , Método Duplo-Cego , Gengivite/tratamento farmacológico , Gengivite/prevenção & controle , Humanos , Método Simples-Cego , Cremes Dentais/uso terapêutico , Resultado do Tratamento , Adulto JovemRESUMO
Acute myeloid leukemia (AML) is the most common hematological malignancy among adults and is characterized by accumulation of immature myeloid cells. Different genetic factors have role in the occurrence of AML. Among different proteins, RUNX1 and BAALC are involved in the development AML. It has been shown that BAALC overexpression is a factor that indicate shorter disease free survival in a subset of AML patients. RUNX1 has been implicated in the development of breast, prostate, lung, and skin cancers. The aim of this study is determination of the prevalence of common polymorphisms in BAALC (rs6999622 and rs62527607) and RUNX1 (rs13051066 and rs61750222) in AML patients compared with healthy subjects. A total of 100 AML patients and 100 healthy control subjects were included in our study. Genomic DNA was isolated from peripheral blood and the polymorphisms were genotyped by applying ARMS and PCR-RFLP methods. Finally, data was analyzed using SPPSS software. Our results demonstrate a significant association between the RUNX1 rs13051066 and AML in the co-dominant (odd ratio = 6.66, 95% Cl = 1.85-25, p = .006) and dominant (GT + TT versus GG: odd ratio = 6.15, 95% CI = 1.73-21.87, p = .002) models. The RUNX1 rs13051066 polymorphism is associated with risk of AML in Iranian population. Future studies should consider larger sample size for assessment of RUNX1 gene polymorphisms, and employ cytogenetic and molecular analyses in AML patients from different ethnic origins.
Assuntos
Subunidade alfa 2 de Fator de Ligação ao Core/genética , Predisposição Genética para Doença , Leucemia Mieloide Aguda/genética , Mutação/genética , Feminino , Estudos de Associação Genética , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único/genéticaRESUMO
Acute myeloid leukemia (AML) is a complex hematological neoplasm with poor prognosis. At present, overwhelming evidence indicates that different genetic abnormalities are relevant to the pathogenesis of AML. Nevertheless, its exact molecular mechanism is still unknown. Recently, it was reported that lncRNAs play crucial roles in tumorigenesis. But, their role in the molecular pathogenesis of AML has not been extensively explored. GAS5, one of the earliest known lncRNAs, has an essential role in the formation and progression of multiple human cancers. It was recently demonstrated that GAS5 acts as a riborepressor of the Glucocorticoid receptor) GR) and abnormal levels of GAS5 may alter response of hematopoietic cells to glucocorticoids. GAS5 can have interaction with the GR that encoded by NR3C1 gene and inhibit its transcriptional activity. To test whether the genetic variants can be associated with AML risk, we genotyped rs55829688 (T > C) polymorphism in GAS5 and three NR3C1 SNPs namely rs6195, rs41423247 and rs6189/rs6190 in a population of 100 Iranian AML patients and 100 healthy subjects. The analysis of the data showed the frequency of alleles and genotypes of rs55829688 and rs6189/rs6190 polymorphisms did not differ between patients and healthy subjects. But, rs41423247 and rs6195 demonstrated a significant correlation with AML risk. The rs6195 was associated with higher AML susceptibility in the co-dominant (OR = 4.58, 95% CI = 2.11-9.981, P < .0001), dominant (OR = 4.55, 95% CI = 2.155-9.613, P < .0001), and over-dominant (OR = 4.43, 95% CI = 2.042-9.621, P < .0001) models. Also, the rs41423247 polymorphism was associated with higher risk of AML in co-dominant (OR = 2.07, 95% CI = 1.171-4.242, P = .012) and dominant (OR = 2.47, 95% CI = 1.192-5.142, P = .010) models. Furthermore, haplotype analysis (rs41423247, rs6189.rs6190, rs6195, and rs55829688 respectively) demonstrated that GGAT, CGGT, and GGGT haplotypes were associated with higher risk of AML in the studied population (p-values = .007, 0.042 and 0.044, respectively). The present study reveals a possible role for NR3C1 in the pathogenesis of AML.
Assuntos
Predisposição Genética para Doença , Leucemia Mieloide/genética , RNA Longo não Codificante/genética , Receptores de Glucocorticoides/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Alelos , Criança , Pré-Escolar , Feminino , Estudos de Associação Genética , Genótipo , Haplótipos/genética , Humanos , Irã (Geográfico)/epidemiologia , Leucemia Mieloide/patologia , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único/genéticaRESUMO
Pompe disease (PD) is a rare autosomal recessive multi-systemic lysosomal storage disorder, caused by mutations in the acid alpha-glucosidase (GAA) gene located on 17q25.2-q25.3. It is one of about 50 rare genetic diseases categorized as lysosomal storage disorders. This disease is characterized by a range of different symptoms related to acid alpha-glucosidase deficiency. Mutation recognition in the GAA gene can be very significant for purposes such as therapeutic interference, early diagnosis and genotype-phenotype relationship. In the current study, peripheral blood samples were gathered from patients with PD and healthy members of three families. Enzymatic activity of GAA was checked. Then, mutation detection was performed by polymerase chain reaction followed by direct sequencing of all exons in samples with decreased enzyme activity. The identified mutations were investigated using bioinformatics tools to predict possible effects on the protein product and also to compare the mutated sequence with near species. Three novel mutations (c.1966-1968delGAG, c.2011-2012delAT and c.1475-1481dupACCCCAC) were identified in the GAA gene. Assessment of the effects of these mutations on protein structure and function showed the possibility of harmful effects and their significant alterations in the protein structure. The three novel GAA gene mutations detected in this study expand the information about the molecular genetics of PD and can be used to helpdiagnosis and genetic counseling of affected families.
Assuntos
Doença de Depósito de Glicogênio Tipo II/genética , Mutação , alfa-Glucosidases/genética , Adolescente , Criança , Pré-Escolar , Análise Mutacional de DNA , Feminino , Predisposição Genética para Doença , Glicogênio/metabolismo , Doença de Depósito de Glicogênio Tipo II/metabolismo , Humanos , Lactente , Masculino , Turquia , alfa-Glucosidases/metabolismoRESUMO
Introduction: Multiple sclerosis (MS) as a progressive chronic disease of the central nervous system (CNS) is characterized by demyelination and axonal loss. Results of genetic studies and clinical trials have proved a key role for the immune system in the pathogenesis of MS. Glucocorticoids (GR) are regarded as potent therapeutic compounds for autoimmune and inflammatory diseases which act through their receptors encoded by Nuclear Receptor Subfamily 3 Group C Member 1 (NR3C1) gene. Meanwhile, the long non-coding RNA (lncRNA) growth arrest specific 5 (GAS5) interacts with GR through binding to the DNA-binding domain (DBD) region and reduces GR transcriptional activity.Methods: The purpose of our study was to evaluate the association between MS and polymorphisms within NR3C1 (rs6189/6190, rs56149945, rs41423247) and GAS5 (rs55829688) genes in 300 relapsing-remitting MS patients and 300 healthy subjects.Results: We demonstrated significant differences in distribution of genotype, allele and haplotype frequencies of rs6189, rs41423247 and rs55829688 between the study groups.Conclusion: Our data may suggest that rs6189, rs41423247 and rs55829688 are associated with the increased risk of MS development. Future studies are needed to verify our results in larger sample sizes and elaborate the underlying mechanisms for contribution of these variants in MS disease.
Assuntos
Esclerose Múltipla/genética , RNA Longo não Codificante/genética , Receptores de Glucocorticoides/genética , Adulto , Feminino , Estudos de Associação Genética , Predisposição Genética para Doença , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Fatores de RiscoRESUMO
Alzheimer's disease (AD) is a heterogeneous disorder with multiple patterns of clinical manifestations. Recently, due to the advance of linkage studies, next-generation sequencing and genome-wide association studies, a large number of putative risk genes for AD have been identified using acquired genome mega data. The genetic association between three causal genes, including amyloid precursor protein, presenilin1, and presenilin2 in early-onset AD (EOAD), was discovered over the past few decades. These discoveries showed that there should be additional genetic risk factors for both EOAD and late-onset AD (LOAD) to help fully explain the leading molecular mechanisms in a single pathophysiological entity. This study reviews the clinical features and genetic etiology of LOAD and discusses a variety of AD-mediated genes that are involved in cholesterol and lipid metabolism, endocytosis, and immune response according to their mutations for more efficient selection of functional candidate genes for LOAD. New mechanisms and pathways have been identified as a result.
Assuntos
Doença de Alzheimer/classificação , Doença de Alzheimer/genética , Predisposição Genética para Doença , Colesterol/genética , Colesterol/metabolismo , Regulação da Expressão Gênica , HumanosRESUMO
Multiple Sclerosis (MS) is a chronic inflammatory disease causing demyelination and neurodegeneration in the central nervous system (CNS). Although the exact etiology of MS is still unclear, both genetic and environmental elements are regarded as causative factors. Environmental factors can induce a cascade of events in immune system leading to neuronal death and nerve demyelination. This paper aims to compare the peripheral transcript levels of Ermin (ERMN) (a gene with putative role in cytoskeletal rearrangements during myelinogenesis) and Listerin E3 Ubiquitin Protein Ligase 1 (LTN1) (a gene with functions in regulating innate immune system) between relapsing-remitting MS (RR-MS) patients and healthy controls. The results showed a significant decrease in ERMN expression (p = 0.022); whereas, no significant difference was detected in LTN1 expression between two groups (p = 0.935). The reduction in ERMN expression in leukocytes could be the cause of demyelinating process in RR-MS patients. Current findings might also have practical importance in prognosis and targeted therapies.
Assuntos
Regulação para Baixo , Leucócitos/metabolismo , Esclerose Múltipla Recidivante-Remitente/genética , Proteínas da Mielina/genética , Ubiquitina-Proteína Ligases/genética , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla Recidivante-Remitente/metabolismo , Proteínas da Mielina/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Adulto JovemRESUMO
A new Schiff base grafted graphene oxide-magnetic chitosan was utilized as a novel sorbent for extraction and quantification of lead ion in blood samples via dispersive magnetic solid phase extraction. The prepared nanocomposite sorbent was characterized by SEM, TEM, FT-IR, XRD, VSM and EDX and the quantification analysis was performed by microsampling flame atomic absorption spectrometry. The important parameters on the extraction efficiency were thoroughly optimized by means of experimental design. Under the optimized conditions, an aliquot of 50â¯mL of sample (pH 6.3) was extracted utilizing 60â¯mg of magnetic nanoparticles during 30â¯min. The sorbent was afterward desorbed using 1.0â¯mL of 0.8â¯molâ¯L-1 HNO3 under fierce vortex for 6â¯min. A preconcentration factor of 20 and an absolute recovery of 40% were provided by the proposed method. The limits of detection (3S/N) and quantification (10â¯S/N) were 0.06â¯ngâ¯mL-1 and 2.0â¯ngâ¯mL-1, respectively. An excellent linearity was achieved within the range of (10-800) ng mL-1 and the regression coefficient was 0.9903. The intra- and inter-day RSDs% were found to be 1.8% and 7.0%, respectively. Furthermore, the method was applied for analysis of blood samples and good accuracies within the range of 97%-108% were obtained.
Assuntos
Quitosana/química , Grafite/química , Chumbo/sangue , Magnetismo , Bases de Schiff/química , Concentração de Íons de Hidrogênio , Limite de Detecção , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Reprodutibilidade dos Testes , Análise Espectral/métodos , Difração de Raios XRESUMO
Alteration in gene expression levels underlies many of the phenotypic differences across species. Because of their highly mutable nature, proximity to the +1 transcription start site (TSS), and the emerging evidence of functional impact on gene expression, core promoter short tandem repeats (STRs) may be considered an ideal source of variation across species. In a genome-scale analysis of the entire Homo sapiens protein-coding genes, we have previously identified core promoters with at least one STR of ≥ 6-repeats, with possible selective advantage in this species. In the current study, we performed reverse analysis of the entire Homo sapiens orthologous genes in mouse in the Ensembl database, in order to identify conserved STRs that have shrunk as an evolutionary advantage to humans. Two protocols were used to minimize ascertainment bias. Firstly, two species sharing a more recent ancestor with Homo sapiens (i.e. Pan troglodytes and Gorilla gorilla gorilla) were also included in the study. Secondly, four non-primate species encompassing the major orders across Mammals, including Scandentia, Laurasiatheria, Afrotheria, and Xenarthra were analyzed as out-groups. We introduce STR evolutionary events specifically identical in primates (i.e. Homo sapiens, Pan troglodytes, and Gorilla gorilla gorilla) vs. non-primate out-groups. The average frequency of the identically shared STR motifs across those primates ranged between 0.00005 and 0.06. The identified genes are involved in important evolutionary and developmental processes, such as normal craniofacial development (TFAP2B), regulation of cell shape (PALMD), learning and long-term memory (RGS14), nervous system development (GFRA2), embryonic limb morphogenesis (PBX2), and forebrain development (APAF1). We provide evidence of core promoter STRs as evolutionary switch codes for primate speciation, and the first instance of identity-by-descent for those motifs at the interspecies level.
Assuntos
Evolução Biológica , Especiação Genética , Repetições de Microssatélites/genética , Primatas/genética , Regiões Promotoras Genéticas , Animais , Bases de Dados Genéticas , Cães , Genoma , Gorilla gorilla/genética , Humanos/genética , Mamíferos/genética , Camundongos , Pan troglodytes/genéticaRESUMO
For the first time, ion-pair based emulsification liquid phase microextraction coupled with a novel approach for phase separation followed by high performace liquid chromatgraphy (HPLC) was utilized for trace determination of sulfonamides in water samples. After the formation of ion-pair complex with a cationic surfactant, sulfonamides were extracted into the drops of dispersed organic extracting solvent. Then, the cloudy solution was passed through an in-line filter located in a suitable holder and was separated based on emulsion filtration. By changing the HPLC valve position, the filter was laid in the mobile phase path, and the extraction phase was eluted by the mobile phase and introduced into the separation column for analysis. The effects of important parameters, such as type of extraction solvent, type of ion-pair agent and its concentration, pH of sample solution, ionic strength, and volume of extraction phase, on the extraction efficiency, were investigated and optimized. Under optimal conditions, the linear range, limits of detection, and precision (relative standard deviations) were 0.3-100, 0.1-0.3 µg L(-1), and 4.7-5.8%, respectively. Preconcentration factors (PFs) for the compounds studied were obtained in the range of 268-664. These PFs correspond to extraction recoveries in the range of 41-97%. The sample throughput of the method was 3 samples per hour, regarding 20 min analysis time for a single procedure. Finally, the method was successfully applied to determine the selected sulfonamides in some water samples.
Assuntos
Monitoramento Ambiental/métodos , Sulfonamidas/análise , Poluentes Químicos da Água/análise , Cromatografia Líquida de Alta Pressão/métodos , Limite de Detecção , Microextração em Fase Líquida , Solventes/química , Sulfanilamida , Sulfanilamidas , Tensoativos/químicaRESUMO
In the present work, a simple and portable analysis device was designed for the first time for the determination of lead ions as the model analyte. The basis of the lead analysis is its extraction and pre-concentration in an acceptor droplet via the application of an electrical field. The acceptor droplet is a KI solution and therefore, the formation of a yellow precipitation of PbI2 was a sign of the presence of lead ion in the solution. Following this, digital picture of the final acceptor droplet was analyzed by investigating its Red-Green-Blue (RGB) components. The results show that the RGB intensities of the acceptor phase are proportionate to the lead concentration in the sample solution. Also, a 9.0 V battery was used to apply the electrical field, and other effective parameters, such as the type of organic liquid membrane, pH of the sample solution, and the extraction time, were considered to obtain the optimal conditions. The model analyte was determined by extracting it from a 100 µL sample solution across a thin layer of 1-octanol, immobilized in the pores of a polypropylene membrane sheet, and into the acceptor droplet via applying a 9.0 V electrical potential for 20 min. The device is capable of determining lead ions down to 20.0 ng mL(-1), with admissible repeatability and reproducibility (the intra- and inter-assay precision ranged between 3.8-7.0% and 9.8-11.9%, respectively). Also, we calculated error% for the model analyte in the range of -8.5 to +4.5, which suggests that the chip offers acceptable accuracy for the analysis of lead ions. The linearity was studied in the range of 50.0-1500 ng mL(-1), with a correlation coefficient of 0.9994. Finally, the designed device was used for the analysis of lead in real samples.
Assuntos
Eletricidade , Dispositivos Lab-On-A-Chip , Concentração de Íons de Hidrogênio , Padrões de ReferênciaRESUMO
A new type of liquid-phase microextraction based on two immiscible organic solvents was optimized and validated for the quantification of lidocaine, ketamine, and cocaine in human urine samples. A hollow-fiber based microextraction technique followed by gas chromatography coupled with mass spectrometry detection was used to reduce matrix interferences and improve limits of detection. The analytes were extracted from aqueous sample with pH 11.0, into a thin layer of organic solvent (n-dodecane) sustained in the pores of a hollow fiber, and then into a second organic acceptor (acetonitrile) located inside the lumen of the hollow fiber. With the application of optimized values, good linearity was obtained in the range of 1-500 µg/L for lidocaine and ketamine and 2-500 µg/L for cocaine with the determination coefficient values (r(2) ) >0.9943. The preconcentration factors and limits of detection (S/N > 3) were 250-350 and 0.01-0.05 µg/L, respectively. Intra and interassay precision values were <7.3 and 9.3%, respectively. The method was successfully applied for the determination and quantification of target analytes in human urine samples.
Assuntos
Anestésicos/urina , Cromatografia Gasosa/métodos , Cocaína/urina , Ketamina/urina , Lidocaína/urina , Microextração em Fase Líquida/métodos , Espectrometria de Massas/métodos , Anestésicos/isolamento & purificação , Cocaína/isolamento & purificação , Humanos , Ketamina/isolamento & purificação , Lidocaína/isolamento & purificação , Microextração em Fase Líquida/instrumentaçãoRESUMO
RNA sequencing (RNA-seq) has undergone substantial advancements in recent decades and has emerged as a vital technique for profiling the transcriptome. The transition from bulk sequencing to single-cell and spatial approaches has facilitated the achievement of higher precision at cell resolution. It provides valuable biological knowledge about individual immune cells and aids in the discovery of the molecular mechanisms that contribute to the development of autoimmune diseases. Celiac disease (CeD) is an autoimmune disorder characterized by a strong immune response to gluten consumption. RNA-seq has led to significantly advanced research in multiple fields, particularly in CeD research. It has been instrumental in studies involving comparative transcriptomics, nutritional genomics and wheat research, cancer research in the context of CeD, genetic and noncoding RNA-mediated epigenetic insights, disease monitoring and biomarker discovery, regulation of mitochondrial functions, therapeutic target identification and drug mechanism of action, dietary factors, immune cell profiling and the immune landscape. This review offers a comprehensive examination of recent RNA-seq technology research in the field of CeD, highlighting future challenges and opportunities for its application.
Assuntos
Doença Celíaca , Doença Celíaca/genética , Doença Celíaca/imunologia , Humanos , Análise de Sequência de RNA/métodos , Transcriptoma , Perfilação da Expressão Gênica/métodos , Epigênese GenéticaRESUMO
BACKGROUND: Alzheimer's disease (AD) is a genetic and sporadic neurodegenerative disease considered by an archetypal cognitive impairment and a decrease in less common cognitive impairment. Notably, the discovery of goals in this paradigm is still a challenge, and understanding basic mechanisms is an important step toward improving disease management. Polyadenylation (PA) and alternative polyadenylation (APA) are two of the most critical RNA processing stages in 3'UTRs that influence various AD-related genes. METHODS: In this study, we assessed Cleavage and polyadenylation specificity factors 1 and 6 (CPSF1 and CPSF6), cleavage stimulation factor 1 (CSTF1), and WD Repeat Domain 33 (WDR33) genes expression in the periphery of 50 AD patients and 50 healthy individuals with age and gender-matched by quantitative real-time PCR. RESULTS: Comparing AD patients with healthy people using expression analysis revealed a substantial increase in CSTF1 (posterior beta = 0.773, adjusted P-value = 0.042). Significant positive correlations were found between CSTF1 and CPSF1 (r = 0.365, P < 0.001), WDR33 (r = 0.506, P < 0.001), and CPSF6 (r = 0.446, P < 0.001) expression levels. CONCLUSION: Although further research is required to determine their potential contribution to AD, our findings offer a fresh perspective on molecular regulatory pathways associated with AD pathogenic mechanisms associated with PA and APA.