DigitalUM :: Browsing by Subject "Gene expression"

Browsing by Subject "Gene expression"

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Open Access
Aberrant expressions of delta-protocadherins in the brain of Npc1 mutant mice
(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2014) Yan, Xin; Lukas, Jan; Lin, Juntang; Ernst, Mathias; Koczan, Dirk; Witt, Martin; Fuellen, Georg; Wree, Andreas; Rolfs, Arndt; Luo, Jiankai
Niemann-Pick type C1 (NPC1) disease is an autosomal recessive disorder characterized by dysmyelination and neurodegeneration, which can result in the death of patients in early childhood in some cases. Members of the delta-protocadherins (Pcdhs) play important roles in neurogenesis and brain development. In this study, we compared expression profiles of Pcdhs in the brain between wild-type and Npc1 mutant mice from postnatal day (P) 9 onwards by in situ hybridization. Our data show that laminar distribution of some Pcdhs in the cerebral cortex of Npc1 mutated mice is different from that of wild-type mice. Furthermore, expressions of Pcdhs by oligodendrocytes in the corpus callosum and by Purkinje cells and granular cells in the cerebellum are strongly decreased in Npc1 mutated mice at later stages. Taken together, our data suggest that aberrant expression of Pcdhs is a pathological process accompanied by neurodegeneration in Npc1 mutant mice.
Open Access
Bioactive compounds of tomato: Cancer chemopreventive effects and influence on the transcriptome in hepatocytes
(Elsevier, 2018-01-03) Periago, María Jesús; Navarro-González, Inmaculada; García-Alonso, Javier; Tecnología de Alimentos, Nutrición y Bromatología
The mechanisms of the effects of tomato extracts (containing lycopene and phenolic compounds) and these pure bioactive compounds on the viability, apoptosis, cell cycle, and the scavenging of ROS of HepG2 cells were investigated. Simultaneously, a transcriptomic analysis in human hepatocytes from obese patients, after the intake of tomato juice, was carried out using Human Gene Expression Array (Affymetrix). The viability of HepG2 decreased after exposure to tomato extracts and pure compounds, but none showed apoptotic activity; however, lycopene was able to inhibit cell cycle progression in phase S. Tomato extracts and pure compounds were able to decrease the ROS generation, the phenolic compounds being more effective than lycopene. Transcriptomic analysis suggested a involvement in the regulation of the cell cycle and apoptosis pathways after the intake of tomato juice, through the lycopene accumulation. These results showed that tomato juice have potential as functional foods for maintenance of liver health.
Open Access
Characterization and mRNA expression in an unusual odontogenic lesion in a patient with tricho-dento-osseous syndrome
(Murcia : F. Hernández, 2003) Dodds, A.P.; Cox, S.A.; Suggs, C.A.; Boyd, C.; Hart, T.C.; Wright, J.T.; Ruiz, R.
Odontogenic lesions are rare, but can be associated with significant morbidity. While their molecular determinants are unknown, they likely express many genes common to normal odontogenesis. This study evaluated the histology and mRNA expression of an unusual odontogenic lesion in a patient with a confirmed history of tricho-dento-osseous syndrome. Methods: Decalcified, frozen 8 µm sections of the lesion were cut and mounted on glass slides and stained with hematoxylin/eosin for analysis. The expression of multiple genes associated with normal odontogenesis and related pathologies were evaluated by RT-PCR, where possible in samples of the hard and soft tissue components of the lesion. Results: Histological examination showed the lesion had large areas of irregular, dentine-like material, enamel matrix, areas of woven immature bone and multiple fully mineralised tooth crowns. Although most of the gene transcripts were amplified from both samples, some, including DLX3/7 and Collagen I demonstrated differential expression. Conclusions: This study shows the gene expression profile of aberrant odontogenesis with associated odontoma formation is similar to that of normal tooth and the genes expressed in other odontogenic lesions. While the role of altered gene expression in the development of such lesions has previously been postulated from transgenic models, this is the only report of an odontogenic lesion in a patient with TDO, and begins to elucidate possible gene interactions key to its development.
Open Access
Consumption of spinach and tomato modifies lipid metabolism, reducing Hepatic Steatosis in rats
(MDPI, 2020-10-24) Elvira Torales, Laura Inés; Navarro González, Inmaculada; Rodrigo García, Joaquín; Seva Alcaraz, Juan; García Alonso, Javier; Periago Castón, María Jesús; Anatomía y Anatomía Patológica Comparada
Non-alcoholic fatty liver disease (NAFLD) is currently a serious and growing clinical problem in developed and developing countries and is considered one of the most frequent chronic liver diseases in the world. The aim of this study was to evaluate the functionality of dietary carotenoids provided by tomato and spinach in the dietary treatment of steatosis. Twenty-two Sprague-Dawley rats with induced steatosis were grouped into three groups and fed standard diet (CD group) and two experimental diets supplemented with 12.75% (LC12.75 group) and 25.5% (HC25.5 group) of a mixture of spinach and tomato powder. Rats fed carotenoid-rich feeds showed an improvement in the plasma biomarkers of steatosis, with lower levels of glucose, total cholesterol, VLDL, TG, proteins, ALT and AST. Likewise, a decrease in oxidative stress was observed, with a significant reduction of malondialdehyde (MDA) in plasma (up to 54%), liver (up to 51.42%) and urine (up to 78.89%) (p < 0.05) and an increase in plasma antioxidant capacity (ORAC) (up to 73.41%) (p < 0.05). Furthermore, carotenoid-rich diets led to an accumulation of carotenoids in the liver and were inversely correlated with the content of total cholesterol and hepatic triglycerides, increasing the concentrations of MUFA and PUFA (up to 32.6% and 48%, respectively) (p < 0.05). The accumulation of carotenoids in the liver caused the modulation of genes involved in lipid metabolism, and we particularly observed an overexpression of ACOX1, APOA1 and NRIH2 (LXR) and the synthesis of the proteins. This study suggests that dietary carotenoids from spinach and tomato aid in the dietary management of steatosis by reversing steatosis biomarkers.
Open Access
Cyclodextrins increase triterpene production in Solanum lycopersicum cell cultures by activating biosynthetic genes
(MDPI, 2022-10-20) Sabater Jara, Ana Belén; Marín Marín, María Jesús; Almagro, Lorena; Pedreño, María Angeles; Biología Vegetal
In this work, Solanum lycopersicum cv. Micro-Tom suspension-cultured cells were used to analyze the effect of different elicitors including β-cyclodextrins (CD), methyl jasmonate (MJ), β-glucan (Glu) and 3-hexenol (Hex) separately and the combined treatments of CD + MJ, CD + glu and CD + Hex on triterpene compound production after 24, 72 and 96 h. Moreover, we studied the changes induced by elicitors in the expression of key biosynthetic genes to elucidate the regulation of the triterpene biosynthetic pathway. The relative abundance of the triterpene compounds identified in the extracellular medium after elicitation (squalene, fucosterol, avenasterol, β-sitosterol, cycloartenol and taraxasterol) was determined by gas chromatography coupled to mass spectrometry, and the expression level of genes in treated-cells was analyzed by real-time quantitative polymerase chain reaction (qRT-PCR). Results showed that, in CD-treated cells (CD, CD + MJ, CD + Glu, CD + Hex), specialized metabolites were accumulated mainly in the extracellular medium after 72 h of elicitation. Moreover, qRT-PCR analysis revealed that the highest triterpene levels in CD-treated cells (CD, CD + MJ, CD + Glu, CD + Hex) were highly correlated with the expression of cycloartenol synthase, 3-hydroxy-3-methylglutaryl-CoA reductase and squalene epoxidase genes at 24 h of treatment, whereas the expression of sterol methyltransferase was increased at 72 h. According to our findings, CD acts as a true elicitor of triterpene biosynthesis and can promote the release of bioactive compounds from the tomato cells into the extracellular medium. The results obtained provide new insights into the regulation of the triterpene metabolic pathway, which might be useful for implementing metabolic engineering techniques in tomato.
Open Access
Differential response of phenol metabolism associated with antioxidative network in elicited grapevine suspension cultured cells under saline conditions
(MDPI, 2022-02-15) Almagro Romero, Lorena; Calderón, Antonio A.; Pedreño, María Angeles; Ferrer, María Ángeles; Biología Vegetal
Highly productive trans-resveratrol (t-R) grapevine suspension cultured cells (SCC) and two effective elicitors, methyl jasmonate (MJ) and randomly methylated β-cyclodextrins (CDs), were used to analyze the extent to which salt treatments alter the production of bioactive phenolic compounds. The expression/activity profile of the enzymes involved in phenol metabolism and an-tioxidant networks were also studied. A marked extracellular accumulation of phenolic compounds, especially t-R, was found in SCC elicited with CDs and/or MJ under saline conditions. However, the treatments with MJ alone and all those combined with salt favored the intracellular accumulation of catechin and ferulic acid. The salt-induced accumulation of phenolics was correlated with the higher total antioxidant capacity values found in cells, suggesting that cellular redox homeostasis under saline conditions was largely maintained by increasing phenolic compound production. These higher levels of phenolics found in elicited cells under saline conditions fit well with the highest activity of phenylalanine ammonia-lyase. Moreover, antioxidant enzyme activities were boosted in treatments with MJ and/or in those combined with salt and decreased in those treated with CDs. These results suggest a differential response of the antioxidative network to the presence of elicitors under saline conditions.
Open Access
DNA repair mechanisms in mammalian germ cells
(Murcia: F. Hernández, 2011) Ozturk, Saffet; Demir, Necdet
Mammalian germ cells encounter several types of DNA damage. This damage is almost completely repaired in a short period of time to provide the maintenance of genomic integrity. The main repair mechanisms operating in mammalian germline cells are: nucleotide excision repair (NER), base excision repair (BER), mismatch repair (MMR), DNA double strand break repair (DSBR), and post replication repair (PRR). Currently, there are relatively few publications that summarize basic information and new findings on DNA repair mechanisms used in mammalian germ cells. In the present article, we review the studies that discuss repair mechanisms operating in the female and male germ cells. We then survey some of the recent discoveries made in this field.
Open Access
Effect of silk fibroin microparticles on cellular immunity and liver of gilthead seabream (Sparus aurata L.) with and without experimental skin injuries
(Elsevier, ) Albaladejo-Riad, Nora; Espinosa-Ruiz, Cristóbal; Esteban Abad, María Ángeles; Biología celular e Histología
Silk fibroin microparticles (SF) were administered in the diet of gilthead seabream with or without experimental skin wounds to study the effects on cellular immunity and liver. A commercial diet was enriched with different SF contents: 0, 50 and 100 mg kg-1 (control, SF50 and SF100 diet, respectively). The animals were fed for 30 days and half of them were sampled. Similar experimental wounds were then performed to the rest of fish and they continued to be fed the same diet. At 7 days post-wounding, samples were taken from the wounded fish. Cellular immunity was studied on head kidney leucocytes (phagocytosis, respiratory and peroxidase content) and liver status (histological study and gene expression) were studied. Our results showed that experimental wounds affect both cellular immunity (by decreasing leucocyte respiratory burst and peroxidase activity) and altered liver histology (by inducing vascularization and congestion of blood vessels). In addition, it affects the expression of genes indicative of oxidative stress, endoplasmic reticulum stress and apoptosis. More specifically, it was the highest dose of silk fibroin (SF100) that significantly increased the phagocytic capacity of leucocytes as well as vascularization in the liver. In addition, increased expression of antioxidant genes (cat and gsr) and decreased expression of genes related to reticulum endoplasmic stress (grp94 and grp170) and apoptosis (nos and jnk) were detected in these fish fed with SF100 and wounded. In conclusion, fed fish with SF100 had many beneficial effects as cellular immunostimulant and hepatoprotection in wounded fish. Its use could be of great interest for stress management in farmed fish conditions.
Embargo
Effect of terbinafine on the biosynthetic pathway of isoprenoid compounds in carrot suspension cultured cells
(Springer, 2018-04-21) Miras Moreno, Begoña; Almagro, Lorena; Pedreño, María Angeles; Sabater Jara, Ana Belén; Biología Vegetal
Plant sterols are essential components of membrane lipids, which contributing to their fluidity and permeability. Besides their cholesterol-lowering properties, they also have anti-inflammatory, antidiabetic and anticancer activities. Squalene, which is phytosterol precursor, is widely used in medicine, foods and cosmetics due to its anti-tumor, antioxidant and anti-aging activities. Nowadays, vegetable oils constitute the main sources of phytosterols and squalene, but their isolation and purification involve complex extraction protocols and high costs. In this work, Daucus carota cell cultures were used to evaluate the effect of cyclodextrins and terbinafine on the production and accumulation of squalene and phytosterols as well as the expression levels of squalene synthase and cycloartenol synthase genes. D. carota cell cultures were able to produce high levels of extracellular being phytosterols in the presence of cyclodextrins (12 mg/L), these compounds able to increase both the secretion and accumulation of phytosterols in the culture medium. Moreover, terbinafine induced a significant increase in intracellular squalene production, as seen after 168 h of treatment (497.0 ± 23.5 µg g dry weight−1) while its extracellular production only increased in the presence of cyclodextrins.The analysis of sqs and cas gene expression revealed that cyclodextrins did not induce genes encoding enzymes involved in the phytosterol biosynthetic pathway since the expression levels of sqs and cas genes in cyclodextrin-treated cells were lower than in control cells. The results, therefore, suggest that cyclodextrins were only able to release phytosterols from the cells to the extracellular medium, thus contributing to their acumulation. To sum up, D. carota cell cultures treated with cyclodextrins or terbinafine were able to produce high levels of phytosterols and squalene, respectively, and, therefore, these suspension-cultured cells of carrot constitute an alternative biotechnological system, which is at the same time more sustainable, economic and ecological for the production of these bioactive compounds.
Open Access
Evaluating the link between BAFF system gene expression and acute rejection development in kidney transplantation
(MDPI, 2022-07-07) Alfaro, Rafael; Llorente, Santiago; Jiménez Coll, Víctor; Martínez Banaclocha, Helios; Galián, José Antonio; Botella, Carmen; Moya Quiles, María Rosa; Muro Pérez, Manuel; Peña Moral, Jesús de la; Minguela, Alfredo; Legaz Pérez, Isabel; Muro, Manuel; Ciencias Sociosanitarias
B-cell activating factor (BAFF) system signaling is critical for B-cell homeostasis, effector functions, and tolerance maintenance in transplants, but it has not been studied in kidney transplant recipients (KTRs). The aim was to analyze the changes in BAFF system expression in KTRs with/without acute rejection (AR/NAR). The BAFF system expression was analyzed by qPCR in 40 KTRs. A meta-analysis of BAFF system expression and histological renal damage was identified by the Chronic Allograft Damage Index (CADI) and performed from the GEO database. Proliferation-inducing ligand (APRIL) expression increased at three- and six-months post-KT (p = 0.014 and p < 0.001). B-cell maturation antigen (BCMA) expression increased at six-months post-KT (p = 0.038). BAFF expression remained stable in NAR-KTRs, but was increased in CADI concerning the No-CADI group at one year (p = 0.008). BCMA expression increased in the CADI group at one- (p = 0.001) and six-years post-KT (p = 0.024). At three months, the transmembrane activator and calcium modulator interactor (TACI) gene significantly elevated KTRs with DSAs (donor-specific antibody; p = 0.034). KTRs with DSAs significantly increase the B-cell activating factor receptor (R-BAFF; p = 0.021) and TACI (p = 0.018) between pre- and three-month post-KT. Changes in the expression of the BAFF system increase during post-KTR in the development of AR and chronic allograft damage, and could be an important pathological tool to detect and prevent kidney graft outcomes.
Embargo
Gene expression changes in colon tissues from colorectal cancer patients following the intake of an ellagitannin-containing pomegranate extract: a randomized clinical trial
(Elsevier, 2017-01-27) Nuñez-Sánchez, María A.; González Sarrías, Antonio; García Villalba, Rocío; Monedero Saiz, Tamara; García Talavera, Noelia V.; Gómez Sánchez, María B.; Sánchez Álvarez, Carmen; García Albert, Ana M.; Rodríguez Gil, Francisco J.; Ruiz Marín, Miguel; Pastor Quirante, Francisco A.; Martínez Díaz, Francisco; Tomás Barberán, Francisco A.; Espín, Juan Carlos; García Conesa, María Teresa; Oftalmología, Optometría, Otorrinolaringología y Anatomía Patológica
The clinical evidence of dietary polyphenols as colorectal cancer (CRC) chemopreventive compounds is very weak. Verification in humans of tissue-specific molecular regulation by the intake of polyphenols requires complex clinical trials that allow for the procurement of sufficient pre- and postsupplementation tissue samples. Ellagitannins (ETs), ellagic acid (EA) and their gut microbiota-derived metabolites, the urolithins, modify gene expression in colon normal and cancer cultured cells. We conducted here the first clinical trial with 35 CRC patients daily supplemented with 900 mg of an ET-containing pomegranate extract (PE) and evaluated the expression of various CRC-related genes in normal and cancerous colon tissues before (biopsies) and after (surgical specimens) 5–35 days of supplementation. Tissues were also obtained from 10 control patients (no supplementation) that confirmed a large, gene- and tissue-specific interindividual variability and impact of the experimental protocol on gene expression, with some genes induced (MYC, CD44, CDKN1A, CTNNB1), some repressed (CASP3) and others not affected (KRAS). Despite these issues, the consumption of the PE was significantly associated with a counterbalance effect in the expression of CD44, CTNNB1, CDKN1A, EGFR and TYMs, suggesting that the intake of this PE modulated the impact of the protocol on gene expression in a gene- and tissue-specific manner. These effects were not associated with the individuals' capacity to produce specific urolithins (i.e., metabotypes) or the levels of urolithins and EA in the colon tissues and did not reproduce in vitro effects evidencing the difficulty of demonstrating in vivo the in vitro results.
Open Access
Genetic and molecular alterations in rhabdomyosarcoma: mRNA overexpression of MCL1 and MAP2K4 genes
(Murcia : F. Hernández, 2009) Pazzaglia, Laura; Chiechi, Antonella; Conti, Amalia; Gamberi, Gabriella; Magagnoli, Giovanna; Novello, Chiara; Morandi, Luca; Picci, Piero; Mercuri, Mario; Benassi, M.S.
Rhabdomyosarcoma, the most common soft tissue sarcoma in childhood, belongs to the small round cell tumor family and is classified according to its histopathological features as embryonal, alveolar and pleomorphic. In this study we propose to explore genetic alterations involved in rhabdomyosarcoma tumorigenesis and assess the level of mRNA gene expression of controlling survival signalling pathways. For genetic and molecular analysis, array-based comparative genomic hybridization, combined with Real Time PCR using the comparative method, was performed on 14 primary well-characterized human primary rhabdomyosarcomas. Multiple changes affecting chromosome arms were detected in all cases, including gain or loss of specific regions harbouring cancer progression-associated genes. Evaluation of mRNA levels showed in the majority of cases overexpression of MCL1 and MAP2K4 genes, both involved in cell viability regulation. Our findings on rhabdomyosarcoma samples showed multiple copy number alterations in chromosome regions implicated in malignancy progression and indicated a strong expression of MAP2K4 and MCL1 genes, both involved in different biological functions of complicated signalling pathways.
Open Access
Improved biotechnological production of paclitaxel in Taxus media cell cultures by the combined action of coronatine and calix[8]arenes
(Elsevier, 2021-06) Escrich, Ainoa; Almagro Romero, Lorena; Moyano, Elisabeth; Cusido, Rosa M.; Bonfill, Mercedes; Hosseini, Bahman; Palazón, Javier; Biología Vegetal
Paclitaxel (PTX), a widely used anticancer agent, is found in the inner bark of several Taxus species, although at such low levels that its extraction is ecologically unsustainable. Biotechnological platforms based on Taxus sp. cell cultures offer an eco-friendlier approach to PTX production, with yields that can be improved by elicitation. However, the also limited excretion of target compounds from the producer cells to the medium hampers their extraction and purification. In this context, we studied the effect of treating T. media cell cultures with the elicitor coronatine (COR) and calix[8]arenes (CAL), nanoparticles that can host lipophilic compounds within their macrocyclic scaffold. The highest taxane production (103.5 mg.L−1), achieved after treatment with COR (1 μM) and CAL (10 mg.L−1), was 15-fold greater than in the control, and PTX represented 82% of the total taxanes analyzed. Expression levels of the flux-limiting PTX biosynthetic genes, BAPT and DBTNBT, increased after the addition of COR, confirming its elicitor action, but not CAL. The CAL treatment significantly enhanced taxane excretion, especially when production levels were increased by COR; 98% of the total taxanes were found in the culture medium after COR + CAL treatment. By forming complexes with PTX, the nanoparticles facilitated its excretion to the medium, and by protecting cells from PTX toxicity, its intra-and extra-cellular degradation may have been avoided. The addition of COR and CAL to T. media cell cultures is therefore a bio-sustainable and economically viable system to improve the yield of this important anticancer compound.
Embargo
Increased glucosinolate production in Brassica oleracea var. italica cell cultures due to coronatine activated genes involved in glucosinolate biosynthesis
(American Chemical Society, 2018-12-19) Sánchez Pujante, P.J.; Sabater Jara, Ana Belén; Belchi Navarro, Sarai; Pedreño, María Angeles; Almagro, Lorena; Biología Vegetal
In this work, the effect of different elicitors and culture conditions on the production of glucosinolates in broccoli cell cultures was studied. The results showed that 0.5 μM coronatine was the best elicitor for increasing glucosinolate production (205-fold increase over untreated cells after 72 h of treatment). Furthermore, the expression levels of some genes related to the biosynthetic pathway of glucosinolates as well as three Myb transcription factors also have been studied. The highest glucosinolate levels found in coronatine-treated cells were closely correlated with the highest gene expression levels of Cyp79b2, Cyp83b1, St5a, Myb51, and Myb122 after 6 h of treatment. The data shown in this study provide new insight into the key metabolic steps involved in the biosynthesis of glucosinolates, which will be of use for future applications of metabolic engineering techniques in broccoli.
Open Access
Influence of Maternal Supplementation with Vitamins, Minerals, and (or) Protein/Energy on Placental Development and Angiogenic Factors in Beef Heifers during Pregnancy
(MDPI, 2024-03-02) Davila Ruiz, Bethania J; Dahlen, Carl R; McCarthy, Kacie L; Caton, Joel S; Hurlbert, Jennifer L; Baumgaertner, Friederike; Menezes, Ana Clara B; Diniz, Wellison JS; Underdahl, Sarah R; Kirsch, James D; Sedivec, Kevin K; Bochantin, Keri A; Borowicz, Pawel P; Cánovas Bernabé, Sebastián; Reynolds, Lawrence P; Fisiología
Understanding placental vascularity is vital for ensuring the proper nourishment of the fetus and, therefore, a healthy offspring. We aimed to investigate the impact of vitamin and mineral supplementation and/or different rates of body weight gain on placental vascularity in beef heifers. To this end, in the first experiment, heifers were divided into groups that received vitamin and mineral supplementation or did not at least 72 days before breeding. At breeding, they were further divided into low or moderate-weight gain groups, resulting in four different treatments maintained until day 83 of pregnancy when tissue collection was performed. In the second experiment, another group of heifers received a basal diet or a diet with vitamin and mineral supplementation from breeding until parturition. We evaluated placental blood vessel density in both experiments and the placental expression of genes related to blood vessel growth in the first experiment. Results showed that supplementation and the rate of body weight gain during early pregnancy did not significantly affect placental vascularity or the expression of angiogenic factors. On the other hand, placental vascularity measured at parturition was increased in the fetal placenta of the supplemented group. These findings suggest that supplementation with vitamins and minerals throughout all gestation may impact placental function at a later stage of pregnancy.
Open Access
Maternal protein restriction changes structural and metabolic gene expression in the skeletal muscle of aging offspring rats
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Valente, Jéssica Silvino; Perez, Érika Stefani; Zanella, Bruna Tereza Thomazini; Gutierrez de Paula, Tassiana; Alcantara dos Santos, Sérgio Alexandre; Oliveira da Silva Duran, Bruno; Carvalho, Robson Francisco; Justulin, Luis Antonio; de Almeida Fantinatti, Bruno Evaristo; Dal-Pai-Silva, Maeli
Maternal protein restriction affects postnatal skeletal muscle physiology with impacts that last through senility. To investigate the morphological and molecular characteristics of skeletal muscle in aging rats subjected to maternal protein restriction, we used aged male rats (540 days old) born of dams fed a protein restricted diet (6% protein) during pregnancy and lactation. Using morphological, immunohistochemical and molecular analyses, we evaluated the soleus (SOL) and extensor digitorum longus (EDL) muscles, muscle fiber cross-sectional area (CSA) (n=8), muscle fiber frequency (n=5) and the gene expression (n=8) of the oxidative markers (succinate dehydrogenase-Sdha and citrate synthase-CS) and the glycolytic marker (lactate dehydrogenase-Ldha). Global transcriptome analysis (n=3) was also performed to identify differentially regulated genes, followed by gene expression validation (n=8). The oxidative SOL muscle displayed a decrease in muscle fiber CSA (*p<0.05) and in the expression of oxidative metabolism marker Sdha (***p<0.001), upregulation of the anabolic Igf-1 (**p<0.01), structural Chad (**p<0.01), and Fmod (*p<0.05) genes, and downregulation of the Hspb7 (**p<0.01) gene. The glycolytic EDL muscle exhibited decreased IIA (*p<0.05) and increased IIB (*p<0.05) fiber frequency, and no changes in muscle fiber CSA or in the expression of oxidative metabolism genes. In contrast, the gene expression of Chad (**p<0.01) was upregulated and the Myog (**p<0.01) gene was downregulated. Collectively, our morphological, immunohistochemical and molecular analyses showed that maternal protein restriction induced changes in the expression of metabolic, anabolic, myogenic, and structural genes, mainly in the oxidative SOL muscle, in aged offspring rats
Open Access
Mechanism of Dihydrofolate Reductase Downregulation in Melanoma by 3-O-(3,4,5-Trimethoxybenzoyl)-(-)-Epicatechin
(WILEY, 2010-06-01) Sánchez del Campo Ferrer, Luis; Chazarra Parres, Soledad; Montenegro Arce, María Fernanda; Cabezas Herrera, Juan; Rodríguez López, José Neptuno; Bioquímica y Biología Molecular A
In our search to improve the stability and cellular absorption of tea polyphenols, we synthesized 3-O-(3,4,5-trimethoxybenzoyl)-(-)-epicatechin (TMECG), which showed high antiproliferative activity against melanoma. TMECG downregulates dihydrofolate reductase (DHFR) expression in melanoma cells and we detail the sequential mechanisms that result from this even. TMECG is specifically activated in melanoma cells to form a stable quinone methide (TMECG-QM). TMECG-QM has a dual action on these cells. First, it acts as a potent antifolate compound, disrupting folate metabolism and increasing intracellular oxidized folate coenzymes, such as dihydrofolate, which is a noncompetitive inhibitor of dihydropterine reductase, an enzyme essential for tetrahydrobiopterin (H4B) recycling. Such inhibition results in H4B deficiency, endothelial nitric oxide synthase (eNOS) uncoupling and superoxide production. Second, TMECG-QM acts as an efficient superoxide scavenger and promotes intra-cellular H2O2 accumulation. Here, we present evidence that TMECG markedly reduces melanoma H4B and NO bioavailability and that TMECG action is abolished by the eNOS inhibitor N-omega-nitro-L-arginine methyl ester or the H2O2 scavenger catalase, which strongly suggests H2O2-dependent DHFR downregulation. In addition, the data presented here indicate that the simultaneous targeting of important pathways for melanoma survival, such as the folate cycle, H4B recycling, and the eNOS reaction, could represent an attractive strategy for fighting this malignant skin pathology
Open Access
MicroRNA expression changes in kidney transplant: diagnostic efficacy of miR-150-5p as potential rejection biomarker, pilot study
(MDPI, 2021-06-22) Alfaro, Rafael; Legaz Pérez, Isabel; Jiménez Coll, Víctor; El kaaoui El band, Jaouad; Martínez Banaclocha, Helios; Galián, José Antonio; Parrado, Antonio; Mrowiec, Anna; Botella, Carmen; Moya Quiles, María Rosa; Boix, Francisco; Peña Moral, Jesús de la; Minguela, Alfredo; Llorente, Santiago; Muro, Manuel; Ciencias Sociosanitarias
Background: The kidney allograft biopsy is considered the gold standard for rejection diagnosis but is invasive and could be indeterminate. Several publications point to the role of miRNA expression in suggesting its involvement in the acceptance or rejection of organ transplantation. This study aimed to analyze microRNAs involved in the differentiation and activation of B and T lymphocytes from kidney transplant (KT) patients’ peripheral blood leukocytes to be used as biomarkers of acute renal rejection (AR). Methods: A total of 15 KT patients with and without acute rejection (AR/NAR) were analyzed and quantified by miRNA PCR array. A total of 84 miRNAs related to lymphocyte differentiation and activation B and T were studied. The functions and biological pathways were analyzed to predict the potential targets of differential expressed miRNAs. Results: Six miRNA were increased in the AR group (miR-191-5p, miR-223-3p, miR-346, miR-423-5p, miR-574-3p, and miR-181d) and miR-150-5p was increased in the NAR group. In silico studies showed a total of 2603 target genes for the increased miRNAs in AR, while for the decrease miRNA, a total of 1107 target-potential genes were found. Conclusions: Our results show that KT with AR shows a decrease in miR-150-5p expression compared to NAR, suggesting that the decrease in miR-150-5p could be related to an increased MBD6 whose deregulation could have clinical consequences.
Open Access
Molecular alterations of monophasic synovial sarcoma: loss of chromosome 3p does not alter RASSF1 and MLH1 transcriptional activity
(Murcia : F. Hernández, 2006) Pazzaglia, L.; Benassi, M.S.; Ragazzini, P.; Gamberi, G.; Ponticelli, F.; Chiechi, A.; Hattinger, C.M.; Morandi, Luca; Alberghini, M.; Zanella, L.; Picci, P.; Mercuri, Mario
Differential diagnosis of monophasic synovial sarcoma requires the detection of specific biological markers. In this study we evaluated the presence of molecular alterations in 15 monophasic synovial sarcomas. Multiple changes affecting chromosome arms were detected by CGH-array in all microdissected cases available, and an association between gain or loss of specific regions harbouring cancer progression-associated genes and aneuploid status was found. The most frequent alteration was loss of 3p including 3p21.3-p23 region that, however, did not involve the promoter regions of the corresponding genes, RASSF1 and MLH1. Using Real-Time PCR, mRNA levels of both resulted moderately high compared to normal tissue; however, the weak to absent protein expression suggests RASSF1 and MLH1 posttranscription deregulation. Moreover, immunohistochemical analysis revealed that both mesenchymal and epithelial antigens were present in diploid tumours. These findings confirm the genetic complexity of monophasic synovial sarcoma and underline the need to integrate different analyses for a better knowledge of this tumour, essential to investigate new diagnostic and prognostic markers.
Open Access
Molecular and cellular aspects and regulation of intestinal lactase-phlorizin hydrolase
(Murcia : F. Hernández, 2001) Naim, H.Y.
Carbohydrates are hydrolyzed in the intestinal lumen by specific enzymes to monosaccharides before transport across the brush border membrane of epithelial cells into the cell interior. The enzymes implicated in the digestion of carbohydrates in the intestinal lumen are membrane-bound glycoproteins that are expressed at the apical domain of the enterocytes. Absent or reduced activity of one of these enzymes is the cause of disaccharide intolerance and malabsorption, the symptoms of which are abdominal pain, cramps or distention, flatulence, nausea and osmotic diarrhea. Lactose intolerance is the most common intestinal disorder that is associated with an absence or drastically reduced levels of an intestinal enzyme, in this case lactase-phlorizin hydrolase (LPH). The pattern of reduction of activity has been temed late onset of lactase deficiency or adult type hypolactasia. It was thought that the regulation of LPH was posttranslational and was associated with altered structural features of the enzyme. Recent studies, however, suggest that the major mechanism of regulation of LPH is transcriptional. Other forms of lactose intolerance include the rare congenital lactase deficiency and secondary forms, such as those caused by mucosa1 injury, due to infectious gastroenteritis, celiac disease, parasitic infection, drug-induced enteritis and Crohn's disease. This review will shed light on important strucural and biosynthetic aspects of LPH, the role played by particular regions of the LPH protein in its transport, polarized sorting, and function, as well as on the gene expession and regulation of the activity of the enzyme.

Browsing by Subject "Gene expression"

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