Is the Cis-Element CACCC-Box a Master Regulatory Element during Cardiovascular Disease? A Bioinformatics Approach from the Perspective of the Krüppel-like Family of Transcription Factors.

The CACCC-box motif emerges as a pivotal cis-regulatory element implicated in diverse developmental processes and diseases, particularly cardiovascular diseases (CVDs). This study centers on the intricate interplay between the CACCC-box and its binding proteins such as: the Krüppel-Like Family (KLF) of transcription factors as primary effectors in the context of CVDs. Our analysis was through a bioinformatics approach, which revealed significant transcriptional activity among KLF subgroup 2, exhibiting the highest number of interactions focusing on the established roles: pluripotency, cancer, and cardiovascular development and diseases. Our analysis reveals KLF's interactions with GATA4, MEF2C, NKX2.5 and other ~90 potential genes that participate in the regulation of the hypertrophic environment (or CVDs' Environment). Also, the GO analysis showed that genes containing the motif CACCC were enriched for multiple CVDs; in combination with STRING analysis, these results pointed to a link between KLFs and these diseases. The analysis further identifies other potential CACCC-box binding factors, such as SP family members, WT1, VEZF1, and -SALL4, which are implicated in cardiac contraction, remodeling, and inflammation processes.

ZO-2 favors Hippo signaling, and its re-expression in the steatotic liver by AMPK restores junctional sealing.

ZO-2 is a peripheral tight junction (TJ) protein whose silencing in renal epithelia induces cell hypertrophy. Here, we found that in ZO-2 KD MDCK cells, in compensatory renal hypertrophy triggered in rats by a unilateral nephrectomy and in liver steatosis of obese Zucker (OZ) rats, ZO-2 silencing is accompanied by the diminished activity of LATS, a kinase of the Hippo pathway, and the nuclear concentration of YAP, the final effector of this signaling route. ZO-2 appears to function as a scaffold for the Hippo pathway as it associates to LATS1. ZO-2 silencing in hypertrophic tissue is due to a diminished abundance of ZO-2 mRNA, and the Sp1 transcription factor is critical for ZO-2 transcription in renal cells. Treatment of OZ rats with metformin, an activator of AMPK that blocks JNK activity, augments ZO-2 and claudin-1 expression in the liver, reduces the paracellular permeability of hepatocytes, and serum bile acid content. Our results suggest that ZO-2 silencing is a common feature of hypertrophy, and that ZO-2 is a positive regulator of the Hippo pathway that regulates cell size. Moreover, our observations highlight the importance of AMPK, JNK, and ZO-2 as therapeutic targets for blood-bile barrier dysfunction.

An increase in O-GlcNAcylation of Sp1 down-regulates the gene expression of pi class glutathione S-transferase in diabetic mice.

Oxidative stress is a key factor contributing to the development of diabetes complications. Glutathione S-transferases (GSTs) protect against products of oxidative stress by conjugating glutathione to electrophilic substrates, producing compounds that are generally less reactive and more soluble. The expression and activity of GSTs during diabetes have been extensively studied, but little is known about regulation mechanisms of Pi-class GST (GSTP). The aim of the present study was to evaluate how GSTP is regulated in a Streptozotocin (STZ)-induced murine diabetes model. GST activity and GSTP expression were determined in adult male mice diabetized with STZ. Specificity protein 1 (Sp1) expression and O-glycosylation, as well as the role of AP-1 members Jun and Fos in the regulation of GSTP expression, were also assessed. The results showed that GST total activity and GSTP mRNA and protein levels were decreased in the diabetic liver, and returned to normal values after insulin administration. The insulin-mimetic drug vanadate was also able to restore GST activity, but failed to recover GSTP mRNA/protein levels. In diabetic animals, O-glycosylated Sp1 levels were increased, whereas, in insulin-treated animals, glycosylation values were similar to those of controls. After vanadate administration, Sp1 expression levels and glycosylation were lower than those of controls. Our results suggest that hyperglycemia could lead to the observed increase in Sp1 O-glycosylation, which would, in turn, lead to a decrease in the expression of Sp1-dependent GSTP in the liver of diabetic mice.

The consumption of milk supplemented with probiotics decreases the occurrence of caries and the salivary concentration of hßD-3 in children.

OBJECTIVES: This study evaluated the effect of milk supplemented with Lactobacillus rhamnosus SP1 on the occurrence of caries and the salivary concentration of human ß-defensin-3 (hßD-3) in preschool children with high caries risk. MATERIALS AND METHODS: A sample of 42 children was randomly assigned to two groups; children in the intervention group were given 150 mL of milk supplemented with 107 CFU/mL of Lactobacillus rhamnosus SP1, while children in the control group were given standard milk, for 10 months. The occurrence of dental caries was assessed using the International Caries Detection and Assessment System (ICDAS), and the concentration of hßD-3 was measured in unstimulated saliva using an ELISA test at baseline and after the intervention. RESULTS: There was an increase in the number of teeth with carious lesions (dICDAS2-6 mft) in the control group, and this increase was statistically significant (p = 0.0489). The concentration of hßD-3 in saliva from the intervention group decreased from 597.91 to 126.29 pg/mL (p = 0.0061), unlike in the control group, where no change in hßD-3 salivary concentration was found. CONCLUSIONS: These findings showed that regular intake of probiotic-supplemented milk in preschool children with high caries risk decreased the occurrence of caries and the salivary levels of hßD-3. CLINICAL RELEVANCE: Our results suggest the need for developing and implementing probiotic supplementation, as adjuvants to the conventional treatments for caries and allow to considerate the salivary levels of hßD-3 as markers of oral tissue homeostasis.

DNA methylation of the RE-1 silencing transcription factor in peripheral blood mononuclear cells and gene expression of antioxidant enzyme in patients with late-onset Alzheimer disease.

Late-onset Alzheimer disease (LOAD) is the most frequent cause of dementia in elderly adults. However, the factors determining disease onset remain unclear. In the elderly, the activation and expression of the gene encoding RE-1 silencing transcription factor (REST) may be a determinant of neuroprotective mechanisms and good amyloidogenic pathway management. In the present study, the minimal promoter region of REST1 was genetically and epigenetically analyzed in blood samples from 21 subjects with LOAD and 20 cognitively healthy elderly subjects. Genomic DNA was isolated, treated with bisulfite and pyrosequenced, and gene expression was determined using real-time PCR. Notably, subjects with LOAD exhibited hypermethylation and significantly diminished expression of REST1 compared with healthy subjects (p = 0.001). In the LOAD group, the gene expression of CAT, SOD2 and GPX also showed a significant decrease and an increase in malondialdehyde. A docking analysis revealed that the first zinc finger protein Sp1 recognized and bound the methylated sequence in subjects with LOAD differently than the binding observed in control subjects. These results reveal that in patients with LOAD the methylation of specific sites in the promoter sequence of REST suppresses its expression and this could be regulating the decreased expression of CAT, SOD and GPX, besides interfering with the action of transcription factors as Sp1.

Occurrence of Podocotyle Dujardin, 1845 (Opecoelidae, Podocotylinae) in three species of deep-sea macrourids from the Gulf of Mexico and Caribbean Sea with an updated key to species and host-parasite checklist.

Podocotyle pearsei Manter, 1934 is documented from the intestine of Vaillant's grenadier, Bathygadus melanobranchus Vaillant (Macrouridae: Bathygadinae), collected from the northeastern and western Gulf of Mexico from 783-841 m depth. The finding of P. pearsei in B. melanobranchus represents the first originally published report of this genus from this host and the fifth documented host species for P. pearsei. We report three unidentified species of Podocotyle, represented by one individual each, from the intestine of the western Atlantic grenadier, Nezumia atlantica (Parr) (Macrouridae: Macrourinae), and from Bathygadus favosus Goode Bean (Macrouridae: Bathygadinae) found at 637 m, 710 m and 1,143 m depths in the northeastern Gulf of Mexico off Florida and from the Caribbean Sea off Colombia. We provide a checklist of the parasites known from the three macrourid species relevant to this study, comment on the biogeography of the five species of Podocotyle now known from the deep sea and discuss the low host specificity observed across this genus. The high number of fish hosts for Podocotyle (i.e. type hosts include at least 22 piscine families) encompassing a wide phylogenetic diversity and diet makes it unlikely that members of a single genus could evolve such a broad array of life histories (i.e. utilize dissimilar intermediate hosts), and we predict in the future that Podocotyle will be taxonomically divided up. Morphological and especially molecular work is needed for Podocotyle as well as for other digenean genera known to inhabit the deep sea. Podocotyle sp. 1 2 represent the first originally published reports of this genus from N. atlantica while Podocotyle sp. 3 represents the first report of this genus from B. favosus. Podocotyle koshari Nagaty, 1973 is declared a species inquirenda, and a dichotomous key to the 27 species of Podocotyle we recognize is provided.

Expression of specificity protein 1 and collagen I in primary pterygial tissues / Expressão da proteína de especificidade 1 e colágeno I em tecidos pterigiais primários

ABSTRACT Purpose: To determine the expression profiles of the transcription factor specificity protein 1 and collagen I in primary pterygial and normal conjunctival tissues, and to explore the role of specificity protein 1 and collagen I in pterygial development. Methods: The pterygial tissues of 20 patients who underwent resection of primary pterygial tissue in our hospital from June 2016 to December 2017 and the conjunctival tissues of 10 patients with enucleation due to trauma were collected. Reverse transcription quantitative-po lymerase chain reaction and western blot analyses were used to detect the relative expression levels of specificity protein 1 and type I collagen at the mRNA and protein levels. Results: The content of specificity protein 1 and collagen I mRNA and protein was significantly greater in primary pterygial tissue than it was in conjunctival tissue (p<0.05). There was a positive correlation between the mRNA and protein levels of specificity protein 1 and collagen I in primary pterygial tissues (protein: r=1, p<0.05; mRNA: r=1, p<0.05). Conclusion: Specificity protein 1 and collagen I are expressed in normal conjunctival and pterygial tissues, but expression is significantly greater in the latter. Specificity protein 1 and collagen I may be involved in the regulation of the development of primary pterygium.

RESUMO Objetivo: Determinar os perfis de expressão do fator de transcrição da proteína de especificidade 1 e do colágeno I em tecidos pterigiais primários e conjuntivais normais, e explorar o papel da proteína de especificidade 1 e colágeno I no desenvolvimento pterigial. Métodos: Foram coletados os tecidos pterigiais de 20 pacientes submetidos à ressecção de tecido de pterígio primário em nosso hospital no período de junho de 2016 a dezembro de 2017 e os tecidos conjuntivais de 10 pacientes com enucleação por trauma. A reação em cadeia da polimerase quantitativa de transcriptase reversa e a análise de Western blot foram utilizadas para detectar os níveis de expressão relativa da proteína de especificidade 1 e colágeno tipo I nos níveis de mRNA e proteína. Resultados: O conteúdo de especificidade da proteína 1 e do mRNA e proteína do colágeno I foi significativamente maior no tecido de pterígio primário do que no tecido conjuntival (p<0,05). Houve correlação positiva entre os níveis de mRNAs e proteína de especificidade 1 e colágeno I nos tecidos primários do pterígio (proteínas: r=1, p<0,05; mRNA: r=1, p<0,05). Conclusão: A proteína de especificidade 1 e do colágeno I é expressa nos tecidos conjuntivais e pterigiais normais, mas a expressão é significativamente maior no segundo. A especificidade da proteína 1 e do colágeno I pode ser envolvida na regulação do desenvolvimento do pterígio primário.

Transcription Factor Sp1 Regulates the Expression of Calcium Channel α2δ-1 Subunit in Neuropathic Pain.

High voltage-activated (HVA) Ca2+ (CaV) channels are oligomeric complexes formed by an ion-conducting main subunit (Cavα1) and at least two auxiliary subunits (Cavß and CaVα2δ). It has been reported that the expression of CaVα2δ1 increases in the dorsal root ganglia (DRGs) of animals with mechanical allodynia, and that the transcription factor Sp1 regulates the expression of the auxiliary subunit. Hence, the main aim of this work was to investigate the role of Sp1 as a molecular determinant of the exacerbated expression of CaVα2δ-1 in the nerve ligation-induced model of mechanical allodynia. Our results show that ligation of L5/L6 spinal nerves (SNL) produced allodynia and increased the expression of Sp1 and CaVα2δ-1 in the DRGs. Interestingly, intrathecal administration of the Sp1 inhibitor mithramycin A (Mth) prevented allodynia and decreased the expression of Sp1 and CaVα2δ-1. Likewise, electrophysiological recordings showed that incubation with Mth decreased Ca2+ current density in the DRG neurons, acting mostly on HVA channels. These results suggest that L5/L6 SNL produces mechanical allodynia and increases the expression of the transcription factor Sp1 and the subunit CaVα2δ-1 in the DRGs, while Mth decreases mechanical allodynia and Ca2+ currents through HVA channels in sensory neurons by reducing the functional expression of the CaVα2δ-1 subunit.

MicroRNA-382 inhibits cell growth and migration in colorectal cancer by targeting SP1.

BACKGROUND: Emerging evidence showed that microRNAs (miRs) play critical roles in human cancers by functioning as either tumor suppressor or oncogene. MIR-382 was found to function as tumor suppressor in certain cancers. However, the role of MIR-382 in colorectal cancer (CRC) is largely unknown. Specificity protein 1 (SP1) is highly expressed in several cancers including CRC and is correlated with poor prognosis, but it is unclear whether or not MIR-382 can regulate the expression of SP1. METHODS: MIR-382 expression level was measured by reverse transcription-quantitative polymerase chain reaction. The connection between MIR-382 and SP1 was validated by luciferase activity reporter assay and western blot assay. Cell counting kit-8 assay and wound-healing assay were conducted to investigate the biological functions of MIR-382 in CRC. RESULTS: In this study, we found MIR-382 expression was downregulated in CRC tissues and cell lines, and the transfection of MIR-382 mimic decreased cell growth and migration. Furthermore, we identified SP1 was a direct target of MIR-382. Overexpression of MIR-382 decreased the expression of SP1, whereas MIR-382 knockdown promoted SP1 expression. We also observed an inversely correlation between MIR-382 and SP1 in CRC tissues. Additionally, we showed that knockdown of SP1 inhibited cell growth and migration and attenuated the effect of MIR-382 inhibitor on cell behaviors. CONCLUSIONS: In conclusion, the present study describes a potential mechanism underlying a MIR-382/SP1 link contributing to CRC development. Thus, MIR-382 may be able to be developed as a novel treatment target for CRC.

Influence of pharmacogenetic polymorphisms and demographic variables on metformin pharmacokinetics in an admixed Brazilian cohort.

AIMS: To identify pharmacogenetic and demographic variables that influence the systemic exposure to metformin in an admixed Brazilian cohort. METHODS: The extreme discordant phenotype was used to select 106 data sets from nine metformin bioequivalence trials, comprising 256 healthy adults. Eleven single-nucleotide polymorphisms in SLC22A1, SLC22A2, SLC47A1 SLC47A2 and in transcription factor SP1 were genotyped and a validated panel of ancestry informative markers was used to estimate the individual proportions of biogeographical ancestry. Two-step (univariate followed by multivariate) regression modelling was developed to identify covariates associated with systemic exposure to metformin, accessed by the area under the plasma concentration-time curve, between 0 and 48 h (AUC0-48h ), after single oral doses of metformin (500 or 1000 mg). RESULTS: The individual proportions of African, Amerindian and European ancestry varied widely, as anticipated from the structure of the Brazilian population The dose-adjusted, log-transformed AUC0-48h 's (ng h ml-1  mg-1 ) differed largely in the two groups at the opposite ends of the distribution histogram, namely 0.82, 0.79-0.85 and 1.08, 1.06-1.11 (mean, 95% confidence interval; P = 6.10-26 , t test). Multivariate modelling revealed that metformin AUC0-48h increased with age, food and carriage of rs12208357 in SLC22A1 but was inversely associated with body surface area and individual proportions of African ancestry. CONCLUSIONS: A pharmacogenetic marker in OCT1 (SLC22A1 rs12208357), combined with demographic covariates (age, body surface area and individual proportion of African ancestry) and a food effect explained 29.7% of the variability in metformin AUC0-48h .

MicroRNA-382 inhibits cell growth and migration in colorectal cancer by targeting SP1

BACKGROUND: Emerging evidence showed that microRNAs (miRs) play critical roles in human cancers by functioning as either tumor suppressor or oncogene. MIR-382 was found to function as tumor suppressor in certain cancers. However, the role of MIR-382 in colorectal cancer (CRC) is largely unknown. Specificity protein 1 (SP1) is highly expressed in several cancers including CRC and is correlated with poor prognosis, but it is unclear whether or not MIR-382 can regulate the expression of SP1. METHODS: MIR-382 expression level was measured by reverse transcription-quantitative polymerase chain reaction. The connection between MIR-382 and SP1 was validated by luciferase activity reporter assay and western blot assay. Cell counting kit-8 assay and wound-healing assay were conducted to investigate the biological functions of MIR-382 in CRC. RESULTS: In this study, we found MIR-382 expression was downregulated in CRC tissues and cell lines, and the transfection of MIR-382 mimic decreased cell growth and migration. Furthermore, we identified SP1 was a direct target of MIR-382. Overexpression of MIR-382 decreased the expression of SP1, whereas MIR-382 knockdown promoted SP1 expression. We also observed an inversely correlation between MIR-382 and SP1 in CRC tissues. Additionally, we showed that knockdown of SP1 inhibited cell growth and migration and attenuated the effect of MIR-382 inhibitor on cell behaviors. CONCLUSIONS: In conclusion, the present study describes a potential mechanism underlying a MIR-382/SP1 link contributing to CRC development. Thus, MIR-382 may be able to be developed as a novel treatment target for CRC.

Probiotic Compared with Standard Milk for High-caries Children: A Cluster Randomized Trial.

The aim of this study was to compare milk supplemented with probiotic lactobacilli with standard milk for the increment of caries in preschool children after 10 mo of intervention. The study was a triple-blind, placebo-controlled randomized trial. Participants were children aged 2 and 3 y (n = 261) attending 16 nursery schools in a metropolitan region in Chile. Nursery schools were randomly assigned to 2 parallel groups: children in the intervention group were given 150 mL of milk supplemented with Lactobacillus rhamnosus SP1 (10(7) CFU/mL), while children in the control group were given standard milk. Interventions took place on weekdays for 10 mo. Data were collected through a clinical examination of participants. The primary outcome measure was the increment of caries in preschool children. This was assessed using the International Caries Detection and Assessment System (ICDAS). The dropout rate was 21%. No differences in caries prevalence were detected between the groups at baseline (P = 0.68). After 10 mo of probiotic intake, the caries prevalence was 54.4% in the probiotic group and 65.8% in the control group. The percentage of new individuals who developed cavitated lesions (ICDAS 5-6) in the control group (24.3%) was significantly higher than that in the probiotic group (9.7%). The increment of dental caries showed an odds ratio of 0.35 (P < 0.05) in favor of the probiotic group. At the cavitated lesion level, the increment of new caries lesions within the groups showed 1.13 new lesions per child in the probiotic group compared with 1.75 lesions in the control group (P < 0.05). The probiotic group showed an increment of 0.58 ± 1.17 new lesions compared with 1.08 ± 1.70 new lesions observed in the control group. The difference in caries increment was significant at the cavitated lesion level (P < 0.01). In conclusion, the regular long-term intake of probiotic-supplemented milk may reduce caries development in high-caries preschool children (ClinicalTrials.gov: NCT01648075).