RESUMEN
The gut microbiota-brain axis allows for bidirectional communication between the microbes in our gastrointestinal (GI) tract and the central nervous system. Psychological stress has been known to disrupt the gut microbiome (dysbiosis) leading to anxiety-like behavior. Pathogens administered into the gut have been reported to cause anxiety. Whether commensal bacteria affect the gut-brain axis is not well understood. In this study, we examined the impact of a commensal sulfate-reducing bacteria (SRB) and its metabolite, hydrogen sulfide (H2S), on anxiety-like behavior. We found that mice gavaged with SRB had increased anxiety-like behavior as measured by the open field test. We also tested the effects of magnesium oxide (MgO) on SRB growth both in vitro and in vivo using a water avoidance stress (WAS) model. We found that MgO inhibited SRB growth and H2S production in a dose-dependent fashion. Mice that underwent psychological stress using the WAS model were observed to have an overgrowth (bloom) of SRB (Deferribacterota) and increased anxiety-like behavior. However, WAS-induced overgrowth of SRB and anxiety-like behavioral effects were attenuated in animals fed a MgO-enriched diet. These findings supported a potential MgO-reversible relationship between WAS-induced SRB blooms and anxiety-like behavior.
RESUMEN
AIMS: Grape seed procyanidin extract (GSE), and milk thistle silymarin extract (MTE) contain structurally distinct polyphenols, and each agent has been shown to exert antineoplastic effects against lung cancer. We hypothesize that combinations of GSE and MTE will additively enhance their anticancer effects against lung cancer. MATERIALS AND METHODS: The anti-proliferative effects of GSE, MTE and combinations were evaluated in lung neoplastic cell lines. A dose range finding (DRF) study to determine safety, bioavailability and bioactivity, followed by human lung cancer xenograft efficacy studies were conducted in female nude mice with once daily gavage of leucoselect phytosome (LP), a standardized GSE, and/or siliphos, a standardized MTE. The roles of tumor suppressors miR-663a and its predicted target FHIT in mediating the additive, anti-proliferative effecs of GSE/MTE were also assessed. KEY FINDINGS: GSE with MTE additively inhibited lung preneoplastic and cancer cell proliferations. Mice tolerated all dosing regimens in the DRF study without signs of clinical toxicity nor histologic abnormalities in the lungs, livers and kidneys. Eight weeks of LP and siliphos additively inhibited lung tumor xenograft growth. Plasma GSE/metabolites and MTE/metabolites showed that the combinations did not decrease systemic bioavailabilities of each agent. GSE and MTE additively upregulated miR-663a and FHIT in lung cancer cell lines; transfection of antisense-miR-663a significantly abrogated the anti-proliferative effects of GSE/MTE, upregulation of FHIT mRNA and protein. LP and siliphos also additively increased miR-663a and FHIT protein in lung tumor xenografts. SIGNIFICANCE: Our findings support clinical translations of combinations of GSE and MTE against lung cancer.
Asunto(s)
Extracto de Semillas de Uva , Neoplasias Pulmonares , MicroARNs , Proantocianidinas , Silimarina , Vitis , Humanos , Femenino , Animales , Ratones , Proantocianidinas/farmacología , Vitis/metabolismo , Silybum marianum , Ratones Desnudos , Extracto de Semillas de Uva/farmacología , Neoplasias Pulmonares/patología , MicroARNs/metabolismoRESUMEN
Grape seed procyanidin extract (GSE) has been shown to exert antineoplastic properties in preclinical studies. Recently, we reported findings from a modified phase I, open-label, dose escalation clinical study conducted to evaluate the safety, tolerability, MTD, and potential chemopreventive effects of leucoselect phytosome, a standardized GSE complexed with soy phospholipids to enhance bioavailability, in heavy active and former smokers. Three months of leucoselect phytosome treatment significantly decreased bronchial Ki-67 labeling index (LI), a marker of cell proliferation on the bronchial epithelium. Because GSE is widely used as a supplement to support cardiovascular health, we evaluate the impact of oral leucoselect phytosome on the fasting serum complex lipid metabolomics profiles in our participants. One month of leucoselect phytosome treatment significantly increased eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), the omega-3 polyunsaturated fatty acids (n-3 PUFA) with well-established anticancer properties. Leucoselect phytosome also significantly increased unsaturated phosphatidylcholines (PC), likely from soy phospolipids in the phytosome and functioning as transporters for these PUFAs. Furthermore, 3-month leucoselect phytosome treatment significantly increased serum prostaglandin (PG) E3 (PGE3), a metabolite of EPA with anti-inflammatory and antineoplastic properties. Such increases in PGE3 correlated with reductions of bronchial Ki-67 LI (r = -0.9; P = 0.0374). Moreover, posttreatment plasma samples from trial participants significantly inhibited proliferation of human lung cancer cell lines A549 (adenocarcinoma), H520 (squamous cell carcinoma), DMS114 (small cell carcinoma), and 1198 (preneoplastic cell line). Our findings further support the potential utility of leucoselect phytosome in reducing cardiovascular and neoplastic risks in heavy former and active smokers. PREVENTION RELEVANCE: In this correlative study of leucoselect phytosome for lung cancer chemoprevention in heavy active and former smokers, we demonstrate for the first time, favorable modulations of n-3PUFA and downstream PGE3 in fasting serum, further supporting the chemopreventive potential of leucoselect phytosome against lung cancer.
Asunto(s)
Extracto de Semillas de Uva/administración & dosificación , Neoplasias Pulmonares/prevención & control , Administración Oral , Alprostadil/análogos & derivados , Alprostadil/sangre , Alprostadil/metabolismo , Bronquios/patología , Línea Celular Tumoral , Ácidos Docosahexaenoicos/sangre , Ácidos Docosahexaenoicos/metabolismo , Ácido Eicosapentaenoico/sangre , Ácido Eicosapentaenoico/metabolismo , Extracto de Semillas de Uva/efectos adversos , Humanos , Neoplasias Pulmonares/sangre , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Resultado del TratamientoRESUMEN
People living in southwest part of United States are exposed to uranium (U) through drinking water, air, and soil. U is radioactive, but independent of this radioactivity also has important toxicological considerations as an environmental metal. At environmentally relevant concentrations, U is both mutagenic and carcinogenic. Emerging evidence shows that U inhibits DNA repair activity, but how U interacts with DNA repair proteins is still largely unknown. Herein, we report that U directly interacts with the DNA repair protein, Protein Poly (ADP-ribose) Polymerase 1 (PARP-1) through direct binding with the zinc finger motif, resulting in zinc release from zinc finger and DNA binding activity loss of the protein. At the peptide level, instead of direct competition with zinc ion in the zinc finger motif, U does not show thermodynamic advantages over zinc. Furthermore, zinc pre-occupied PARP-1 zinc finger is insensitive to U treatment, but U bound to PARP-1 zinc finger can be partially replaced by zinc. These results provide mechanistic basis on molecular level to U inhibition of DNA repair.
Asunto(s)
Reparación del ADN/fisiología , Reparación del ADN/efectos de la radiación , Poli(ADP-Ribosa) Polimerasa-1/metabolismo , Poli(ADP-Ribosa) Polimerasa-1/efectos de la radiación , Uranio/metabolismo , Uranio/toxicidad , Secuencia de Aminoácidos , Células Cultivadas , Exposición a Riesgos Ambientales/efectos adversos , Humanos , Recién Nacido , Queratinocitos/metabolismo , Queratinocitos/efectos de la radiación , Poli(ADP-Ribosa) Polimerasa-1/genética , Unión Proteica/efectos de los fármacos , Unión Proteica/fisiologíaRESUMEN
Cancer mortality is mainly caused by metastasis, which requires dynamic remodeling of cytoskeletal components such as microtubules. Targeting microtubules presents a promising antimetastatic strategy that could prevent cancer spreading and recurrence. It is known that arsenic trioxide (ATO) is able to inhibit the migration and invasion of solid malignant tumors, but its exact molecular mechanism remains unclear. Here, we report a novel molecular target and antimetastatic mechanism of ATO in head and neck squamous cell carcinoma (HNSCC). We found that cytoplasmic linker protein 170 (CLIP170) was overexpressed in HNSCC tissues and cells compared to normal controls. ATO at non-cytotoxic level (1 µM) inhibited the migration and invasion of HNSCC cells by displacing zinc in the zinc finger motif of CLIP170, which is a key protein that controls microtubule dynamics. The antimetastatic effects of ATO were equivalent to those of siRNA-mediated CLIP170 knockdown. Furthermore, ATO dysregulated microtubule polymerization via the CLIP170/LIS1/NDEL1/dynein signaling pathway in Cal27 cells as a functional consequence of CLIP170 zinc finger disruption. The effect was partially reversed by zinc supplementation. Taken together, these findings reveal that CLIP170 is a novel molecular target of ATO and demonstrate the capability and underlying mechanisms of ATO as a potential antimetastatic agent for HNSCC treatment.
Asunto(s)
1-Alquil-2-acetilglicerofosfocolina Esterasa/metabolismo , Trióxido de Arsénico/farmacología , Proteínas Portadoras/metabolismo , Dineínas/metabolismo , Neoplasias de Cabeza y Cuello/metabolismo , Proteínas Asociadas a Microtúbulos/metabolismo , Proteínas de Neoplasias/metabolismo , 1-Alquil-2-acetilglicerofosfocolina Esterasa/genética , Adulto , Proteínas Portadoras/genética , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Dineínas/genética , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Masculino , Proteínas Asociadas a Microtúbulos/genética , Microtúbulos , Persona de Mediana Edad , Proteínas de Neoplasias/genéticaRESUMEN
Grape seed procyanidin extract (GSE) had been reported to exert antineoplastic properties in preclinical studies. A modified phase I, open-label, dose-escalation clinical study was conducted to evaluate the safety, tolerability, MTD, and potential chemopreventive effects of leucoselect phytosome (LP), a standardized GSE complexed with soy phospholipids to enhance bioavailability, in heavy active and former smokers. Eight subjects ages 46-68 years were enrolled into the study and treated with escalating oral doses of LP for 3 months. Bronchoscopies with bronchoalveolar lavage and bronchial biopsies were performed before and after 3 months of LP treatment. Hematoxylin and eosin stain for histopathology grading and IHC examination for Ki-67 proliferative labeling index (Ki-67 LI) were carried out on serially matched bronchial biopsy samples from each subject to determine responses to treatment. Two subjects were withdrawn due to issues unrelated to the study medication, and a total of 6 subjects completed the full study course. In general, 3 months of LP, reaching the highest dose per study protocol was well tolerated and no dosing adjustment was necessary. Such a treatment regimen significantly decreased bronchial Ki-67 LI by an average of 55% (P = 0.041), with concomitant decreases in serum miR-19a, -19b, and -106b, which were oncomirs previously reported to be downregulated by GSE, including LP, in preclinical studies. In spite of not reaching the original enrollment goal of 20, our findings nonetheless support the continued clinical translation of GSE as an antineoplastic and chemopreventive agent against lung cancer.
Asunto(s)
Antineoplásicos Fitogénicos/administración & dosificación , Biflavonoides/administración & dosificación , Catequina/administración & dosificación , Extracto de Semillas de Uva/administración & dosificación , Neoplasias Pulmonares/prevención & control , Proantocianidinas/administración & dosificación , Administración Oral , Anciano , Antineoplásicos Fitogénicos/efectos adversos , Antineoplásicos Fitogénicos/química , Apoptosis , Biflavonoides/efectos adversos , Biflavonoides/química , Biopsia , Bronquios/diagnóstico por imagen , Bronquios/efectos de los fármacos , Bronquios/patología , Broncoscopía , Catequina/efectos adversos , Catequina/química , Proliferación Celular , Regulación hacia Abajo/efectos de los fármacos , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Extracto de Semillas de Uva/efectos adversos , Extracto de Semillas de Uva/química , Humanos , Antígeno Ki-67/análisis , Antígeno Ki-67/metabolismo , Neoplasias Pulmonares/etiología , Neoplasias Pulmonares/genética , Masculino , MicroARNs/sangre , MicroARNs/metabolismo , Persona de Mediana Edad , Proantocianidinas/efectos adversos , Proantocianidinas/química , Fumar/efectos adversos , Resultado del TratamientoRESUMEN
MiR-106b is an oncomir and a potential target for anti-cancer therapy. We hypothesize that grape seed procyanidin extract (GSE) exerts antineoplastic effects on lung cancer through modulations of miR-106b and its downstream target. We found that GSE significantly down-regulated miR-106b in a variety of lung neoplastic cells and increased cyclin-dependent kinase inhibitor 1A (CDKN1A) mRNA and protein (p21) levels. Transfection of miR-106b mimics reversed the up-regulations of CDKN1A mRNA and p21, abrogated the GSE induced anti-proliferative and anti-invasive properties in lung cancer cells. Oral gavage of leucoselect phytosome (LP), a standardized GSE to athymic nude mice down-regulated MIR106B mRNA and miR-106b expressions, and increased CDKN1A mRNA expression in tumor xenografts, correlating to significant reduction of tumor growth. To assess bioavailability, GSE and metabolites in plasma levels, between 60-90 minutes after gavage of LP were measured by LC/MS at treatment week 4 and 8. A novel bioactivity assay was also developed using lung homogenates from treated mice co-cultured with human lung cancer cells. LP-treated mouse lung homogenates significantly reduced proliferations of various lung cancer cells. Our findings reveal novel antineoplastic mechanisms by GSE, further define the pharmacokinetics and pharmacodynamics of LP, and support the continued investigation of LP against lung cancer.
RESUMEN
Grape seed procyanidin extract (GSE) has been reported to exert antineoplastic properties via the inhibition of cyclooxygenase-2 (COX-2)/prostaglandin E2 (PGE2) eicosanoid pathways. In addition, ample data link carcinogenesis to inflammatory events involving other major eicosanoid metabolic pathways, including prostacyclin (PGI2) and 15-hydroxyeicosatetraenoic acid (15-HETE). We therefore evaluated the effects of GSE on prostacyclin synthase (PTGIS)/PGI2 and 15-lipoxigenase-2 (15-LOX-2)/15-HETE productions by human lung premalignant and malignant cells and correlated the findings with antiproliferative or proapoptotic effects of GSE. The effects of GSE on PGI2 and 15-HETE productions by human bronchoalveolar lavage (BAL) cells ex vivo were also determined. We further evaluated the bioactivity of oral administration of leucoselect phytosome (a standardized GSE) in the lungs of subjects participating in a lung cancer chemoprevention trial, by comparing the antiproliferative effects of coculturing matched pre- versus posttreatment BAL fluids with lung premalignant and malignant cells. GSE significantly increased PGI2 (as measured by 6-keto PGF1α) and 15-HETE productions by these cells. Transfections of PTGIS or 15-LOX-2-specific siRNA partially abrogated the antiproliferative or proapoptotic effects of GSE in lung premalignant and malignant cells, respectively. GSE also increased PTGIS and inhibition of caspase-3, and transfection of 15-LOX-2 siRNA abrogated the GSE-induced apoptosis in A549 cells. In addition, culture supernatants from ex vivo GSE-treated baseline BAL cells, as well as BAL fluids from subjects treated with leucoselect phytosome, significantly decreased proliferations of lung premalignant and malignant cells. Our findings support the continued investigation of GSE as an anti-neoplastic and chemopreventive agent against lung cancer. Cancer Prev Res; 9(12); 925-32. ©2016 AACR.
Asunto(s)
Antineoplásicos Fitogénicos/uso terapéutico , Sistema Enzimático del Citocromo P-450/metabolismo , Ácido Eicosapentaenoico/análogos & derivados , Extracto de Semillas de Uva/uso terapéutico , Oxidorreductasas Intramoleculares/metabolismo , Neoplasias Pulmonares/tratamiento farmacológico , Células A549 , Antineoplásicos Fitogénicos/farmacología , Antioxidantes/farmacología , Antioxidantes/uso terapéutico , Apoptosis/efectos de los fármacos , Biflavonoides/farmacología , Biflavonoides/uso terapéutico , Líquido del Lavado Bronquioalveolar , Broncoscopía , Caspasa 3/metabolismo , Catequina/farmacología , Catequina/uso terapéutico , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Técnicas de Cocultivo , Eicosanoides/análisis , Ácido Eicosapentaenoico/metabolismo , Estudios de Factibilidad , Extracto de Semillas de Uva/farmacología , Humanos , Proyectos Piloto , Proantocianidinas/farmacología , Proantocianidinas/uso terapéutico , ARN Interferente Pequeño/metabolismoRESUMEN
Oncomirs are microRNAs (miRNA) associated with carcinogenesis and malignant transformation. They have emerged as potential molecular targets for anti-cancer therapy. We hypothesize that grape seed procyanidin extract (GSE) exerts antineoplastic effects through modulations of oncomirs and their downstream targets. We found that GSE significantly down-regulated oncomirs miR-19a and -19b in a variety of lung neoplastic cells. GSE also increased mRNA and protein levels of insulin-like growth factor II receptor (IGF-2R) and phosphatase and tensin homolog (PTEN), both predicted targets of miR-19a and -19b. Furthermore, GSE significantly increased PTEN activity and decreased AKT phosphorylation in A549 cells. Transfection of miR-19a and -19b mimics reversed the up-regulations of IGF2R and PTEN gene expression and abrogated the GSE induced anti-proliferative response. Additionally, oral administration of leucoselect phytosome, comprised of standardized grape seed oligomeric procyanidins complexed with soy phospholipids, to athymic nude mice via gavage, significantly down-regulated miR-19a, -19b and the miR-17-92 cluster host gene (MIR17HG) expressions, increased IGF-2R, PTEN, decreased phosphorylated-AKT in A549 xenograft tumors, and markedly inhibited tumor growth. To confirm the absorption of orally administered GSE, plasma procyanidin B1 levels, between 60 and 90 min after gavage of leucoselect phytosome (400 mg/kg), were measured by LC/MS at week 2 and 8 of treatment; the estimated concentration that was associated with 50% growth inhibition (IC50) (1.3 µg/mL) in vitro was much higher than the IC50 (0.032-0.13 µg/ml) observed in vivo. Our findings reveal novel antineoplastic mechanisms by GSE and support the clinical translation of leucoselect phytosome as an anti-neoplastic and chemopreventive agent for lung cancer.
Asunto(s)
Antineoplásicos Fitogénicos/uso terapéutico , Carcinoma de Pulmón de Células no Pequeñas/prevención & control , Suplementos Dietéticos , Extracto de Semillas de Uva/uso terapéutico , Neoplasias Pulmonares/prevención & control , MicroARNs/antagonistas & inhibidores , Proantocianidinas/uso terapéutico , ARN Neoplásico/antagonistas & inhibidores , Animales , Antineoplásicos Fitogénicos/metabolismo , Antioxidantes/metabolismo , Antioxidantes/uso terapéutico , Apoptosis , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/patología , Línea Celular Tumoral , Proliferación Celular , Femenino , Extracto de Semillas de Uva/metabolismo , Humanos , Absorción Intestinal , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Ratones Desnudos , MicroARNs/metabolismo , Proteínas de Neoplasias/agonistas , Proteínas de Neoplasias/metabolismo , Proantocianidinas/metabolismo , ARN Neoplásico/metabolismo , Distribución Aleatoria , Carga Tumoral , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Parasitism genes encode effector proteins that are secreted through the stylet of root-knot nematodes to dramatically modify selected plant cells into giant-cells for feeding. The Mi8D05 parasitism gene previously identified was confirmed to encode a novel protein of 382 amino acids that had only one database homolog identified on contig 2374 within the Meloidogyne hapla genome. Mi8D05 expression peaked in M. incognita parasitic second-stage juveniles within host roots and its encoded protein was limited to the subventral esophageal gland cells that produce proteins secreted from the stylet. Constitutive expression of Mi8D05 in transformed Arabidopsis thaliana plants induced accelerated shoot growth and early flowering but had no visible effects on root growth. Independent lines of transgenic Arabidopsis that expressed a double-stranded RNA complementary to Mi8D05 in host-derived RNA interference (RNAi) tests had up to 90% reduction in infection by M. incognita compared with wild-type control plants, suggesting that Mi8D05 plays a critical role in parasitism by the root-knot nematode. Yeast two-hybrid experiments confirmed the specific interaction of the Mi8D05 protein with plant aquaporin tonoplast intrinsic protein 2 (TIP2) and provided evidence that the Mi8D05 effector may help regulate solute and water transport within giant-cells to promote the parasitic interaction.
Asunto(s)
Arabidopsis/parasitología , Proteínas del Helminto/genética , Proteínas de la Membrana/metabolismo , Enfermedades de las Plantas/parasitología , Proteínas de Plantas/metabolismo , Solanum lycopersicum/parasitología , Tylenchoidea/genética , Secuencia de Aminoácidos , Animales , Arabidopsis/genética , Arabidopsis/fisiología , Transporte Biológico , Susceptibilidad a Enfermedades , Femenino , Flores/genética , Flores/parasitología , Flores/fisiología , Expresión Génica , Células Gigantes , Proteínas del Helminto/metabolismo , Interacciones Huésped-Parásitos , Estadios del Ciclo de Vida , Solanum lycopersicum/genética , Solanum lycopersicum/fisiología , Raíces de Plantas/genética , Raíces de Plantas/parasitología , Raíces de Plantas/fisiología , Brotes de la Planta/genética , Brotes de la Planta/parasitología , Brotes de la Planta/fisiología , Plantas Modificadas Genéticamente , Mapeo de Interacción de Proteínas , Interferencia de ARN , Alineación de Secuencia , Técnicas del Sistema de Dos Híbridos , Tylenchoidea/crecimiento & desarrollo , Tylenchoidea/fisiología , Agua/metabolismoRESUMEN
The biosynthesis of female moth sex pheromone blends is controlled by a number of different enzymes, many of which are encoded by members of multigene families. One such multigene family, the acyl-CoA desaturases, is composed of certain genes that function as key players in moth sex pheromone biosynthesis. Although much is known regarding the function of some of these genes, very little is known regarding how novel genes have evolved within this family and how this might impact the establishment of new sex pheromone blends within a species. We have discovered that several cryptic Delta11 and Delta14 desaturase genes exist in the genomes of the European and Asian corn borers (Ostrinia nubilalis and Ostrinia furnacalis, respectively). Furthermore, an entirely novel class of desaturase gene has arisen in the Ostrinia lineage and is derived from duplication of the Delta11 desaturase gene and subsequent fusion with a retroposon. Interestingly, the genes have been maintained over relatively long evolutionary time periods in corn borer genomes, and they have not been recognizably pseudogenized, suggesting that they maintain functional integrity. The existence of cryptic desaturase genes in moth genomes indicates that the evolution of moth sex pheromone desaturases in general is much more complex than previously recognized.
Asunto(s)
Ácido Graso Desaturasas/genética , Duplicación de Gen , Genoma de Planta , Atractivos Sexuales/genética , Zea mays/genética , Animales , Clonación Molecular , Evolución Molecular , Ácido Graso Desaturasas/fisiología , Genes de Plantas , Modelos Genéticos , Datos de Secuencia Molecular , Filogenia , Retroelementos , Atractivos Sexuales/fisiologíaRESUMEN
Experiments were carried out with two strawberry (Fragaria x nanassa Duch.) cultivars Fengxiang and Hongfeng, with different softening characteristics during growth, ripening and postharvest storage. The fruits were harvested at different stages of growth and ripening, as assessed by size and the coloration of the surface of the fruits. We selected the following stages: small and green (S1), large and green (S2), white (S3), reddish (S4), and fully red (S5). The main results were as follows. Both alpha- and beta-galactosidase activities were changed with ripening of strawberry fruits (Fig.1A, B). Of the strawberry cultivars tested, no correlation was found between glucosidase activity and fruit ripening (Fig.1C, D). Alpha-mannosidase is an enzyme being ionically bound with cell wall and its activities is correlated with the softening of strawberry fruits (Fig.1E, F). No beta-mannosidase has been detected in strawberry. The activities of cellulase increased as the strawberry fruits developed from stage of small and green to stage of overripe (Fig.2A). The activities of PME increased during the development of strawberry fruits (Fig.2B). Endo-PG was not detected in strawberry, and exo-PG was not related to fruit ripening (Fig.2C). Changes in cell wall component contents were clearly related to the changes in the firmness of strawberry fruits. The increase in soluble pectin, together with reduction of ionically bound pectin content, covalently bound pectin content (Fig.3A, B) and cellulose (Fig.3A, B) resulted in softening of strawberry fruits.
Asunto(s)
Pared Celular/metabolismo , Fragaria/enzimología , Frutas/enzimología , Fragaria/crecimiento & desarrollo , Fragaria/metabolismo , Frutas/crecimiento & desarrollo , Frutas/metabolismo , Glicósido Hidrolasas/metabolismo , Pectinas/metabolismo , Factores de Tiempo , alfa-Manosidasa/metabolismo , beta-Manosidasa/metabolismoRESUMEN
Six acyl-CoA desaturase-encoding cDNAs from mRNA isolated from the spotted fireworm moth, Choristoneura parallela (Lepidoptera: Tortricidae) were characterized and assayed for functionality. The expression levels of these cDNAs were determined in the pheromone gland and fat body by real-time PCR and the resulting patterns are in line with results from published studies on other moth sex pheromone desaturases. The cDNAs were found to correspond to six genes. Using both biochemical and phylogenetic analyses, four of these were found to belong to previously characterized desaturase functional groups [the Delta 10,11, the Delta 9 (16>18) and the Delta 9 (18>16) groups]. A desaturase highly expressed in the pheromone gland was a novel E11 desaturase that was specific to 14-carbon precursor acids. The fifth gene [CpaZ9(14-26)] was found to display a novel Z9 activity indicating that it belongs to a new Delta 9 functional group, whereas the sixth gene was determined to be nonfunctional with respect to desaturase activity. In accordance with previous studies, we find that desaturases of the Delta 10,11 and Delta 14 groups, which are the fastest evolving desaturases and possess the novel pheromone biosynthetic function, are expressed primarily in the pheromone gland whereas all other desaturases, which do not possess the novel reproductive function, evolve more slowly and display the ancestral metabolic function and pattern of gene expression.