RESUMEN
Mulberry is a rapidly growing plant that thrives in diverse climatic, topographical, and soil types, spanning temperature and temperate countries. Mulberry plants are valued as functional foods for their abundant chemical composition, serving as a significant reservoir of bioactive compounds like proteins, polysaccharides, phenolics, and flavonoids. Moreover, these compounds displayed potent antioxidant activity by scavenging free radicals, inhibiting reactive oxygen species generation, and restoring elevated nitric oxide production induced by LPS stimulation through the downregulation of inducible NO synthase expression. Active components like oxyresveratrol found in Morus demonstrated anti-inflammatory effects by inhibiting leukocyte migration through the MEK/ERK signaling pathway. Gallic and chlorogenic acids in mulberry leaves (ML) powder-modulated TNF, IL-6, and IRS1 proteins, improving various inflammatory conditions by immune system modulation. As we delve deeper into understanding its anti-inflammatory potential and how it works therapeutically, it is crucial to refine the extraction process to enhance the effectiveness of its bioactive elements. Recent advancements in extraction techniques, such as solid-liquid extraction, pressurized liquid extraction, superficial fluid extraction, microwave-assisted extraction, and ultrasonic-assisted extraction, are being explored. Among the extraction methods tested, including Soxhlet extraction, maceration, and ultrasound-assisted extraction (UAE), UAE demonstrated superior efficiency in extracting bioactive compounds from mulberry leaves. Overall, this comprehensive review sheds light on the potential of mulberry as a natural immunomodulatory agent and provides insights into its mechanisms of action for future research and therapeutic applications.
Asunto(s)
Morus , Extractos Vegetales , Morus/química , Extractos Vegetales/farmacología , Extractos Vegetales/química , Humanos , Animales , Antiinflamatorios/farmacología , Antiinflamatorios/química , Agentes Inmunomoduladores/farmacología , Agentes Inmunomoduladores/química , Factores Inmunológicos/farmacología , Factores Inmunológicos/química , Hojas de la Planta/químicaRESUMEN
BACKGROUND: Limosilactobacillusmucosae (LM) exerts anti-inflammatory and health-promoting effects. However, its role in the modulation of gut serotonin or 5-hydroxytryptamine (5-HT) metabolism and 5-HT receptors (HTRs) in inflammation requires further investigation. OBJECTIVES: We compared LM with Lactobacillus amylovorus (LA) for the regulation of 5-HT, HTRs, inflammatory mediators, and their correlations in the colon of mice with experimental colitis. METHODS: Male C57BL/6 mice were randomly assigned to 6 groups: control (Con), LM, LA, dextran sodium sulfate (DSS), and DSS with pre-administration of LM (+LM) or LA (+LA). After 7 d of DSS treatment, mice were killed to analyze the expression of inflammatory mediators, HTRs, and concentrations of 5-HT and microbial metabolites in the colon. RESULTS: LM was more effective than LA in alleviating DSS-induced colonic inflammation. Compared with mice in the DSS group, mice receiving DSS + LM or DSS + LA treatment had lower (P < 0.05) colonic mRNA expression of proinflammatory cytokines. DSS + LM treatment had lower mRNA expression of Il1b, Tnfa, and Ccl3, an abundance of p-STAT3, and greater expression of Tgfb2 and Htr4 in the colon (P < 0.05). The expression of inflammatory mediators (including Tgfb-1) was positively correlated (P < 0.05) with 5-HT and Htr2a and negatively correlated (P < 0.05) with Htr4. However, the expression of Tgfb-2 showed reversed correlations with the 5-HT and HTRs described above. Patterns for these correlations were different for LM and LA. Mice receiving the DSS + LM treatment had greater (P < 0.05) concentrations of acetate and valerate and lower (P < 0.05) concentrations of indole-3-acetic acid in the cecal and colonic contents. CONCLUSIONS: LM showed greater efficacy than LA in alleviating DSS-induced colonic inflammation. The coordinated regulation of transforming growth factor-ß subtypes and serotonin receptors in the colon may be one of the most important mechanisms underlying the probiotic effects of lactobacilli in gut inflammation.
Asunto(s)
Colitis , Serotonina , Masculino , Animales , Ratones , Serotonina/metabolismo , Lactobacillus acidophilus/metabolismo , Regulación hacia Arriba , Ratones Endogámicos C57BL , Colitis/inducido químicamente , Colitis/prevención & control , Colitis/metabolismo , Colon/metabolismo , Inflamación/metabolismo , ARN Mensajero/metabolismo , Factores de Crecimiento Transformadores/efectos adversos , Factores de Crecimiento Transformadores/metabolismo , Sulfato de Dextran/toxicidad , Modelos Animales de EnfermedadRESUMEN
Recently, the drawbacks arising from the overuse of antibiotics have drawn growing public attention. Among them, drug-resistance (DR) and even multidrug-resistance (MDR) pose significant challenges in clinical practice. As a representative of a DR or MDR pathogen, Staphylococcus aureus can cause diversity of infections related to different organs, and can survive or adapt to the diverse hostile environments by switching into other phenotypes, including biofilm and small colony variants (SCVs), with altered physiologic or metabolic characteristics. In this review, we briefly describe the development of the DR/MDR as well as the classical mechanisms (accumulation of the resistant genes). Moreover, we use multidimensional scaling analysis to evaluate the MDR relevant hotspots in the recent published reports. Furthermore, we mainly focus on the possible non-classical resistance mechanisms triggered by the two important alternative phenotypes of the S. aureus, biofilm and SCVs, which are fundamentally caused by the different global regulation of the S. aureus population, such as the main quorum-sensing (QS) and agr system and its coordinated regulated factors, such as the SarA family proteins and the alternative sigma factor σB (SigB). Both the biofilm and the SCVs are able to escape from the host immune response, and resist the therapeutic effects of antibiotics through the physical or the biological barriers, and become less sensitive to some antibiotics by the dormant state with the limited metabolisms.
Asunto(s)
Antibacterianos/farmacología , Proteínas Bacterianas/metabolismo , Biopelículas/crecimiento & desarrollo , Farmacorresistencia Bacteriana/genética , Infecciones Estafilocócicas/microbiología , Staphylococcus aureus/genética , Animales , Proteínas Bacterianas/genética , Biopelículas/efectos de los fármacos , Regulación Bacteriana de la Expresión Génica , Humanos , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/patogenicidadRESUMEN
Intestinal inflammatory disorders, such as inflammatory bowel disease, are major contributors to mortality and morbidity in humans and animals worldwide. While some native peptides have great potential as therapeutic agents against intestinal inflammation, potential cytotoxicity, anti-inciting action, and suppression of anti-inflammatory activity may limit their development as anti-inflammatory agents. Peptide hybridization is an effective approach for the design and engineering of novel functional peptides because hybrid peptides combine the advantages and benefits of various native peptides. In the present study, a novel hybrid anti-inflammatory peptide that combines the active center of Cecropin A (C) and the core functional region of LL-37 (L) was designed [C-L peptide; C (1-8)-L (17-30)] through in silico analysis to reduce cytotoxicity and improve the anti-inflammatory activity of the parental peptides. The resulting C-L peptide exhibited lower cytotoxicity than either C or L peptides alone. C-L also exerted a protective effect against lipopolysaccharide (LPS)-induced inflammatory responses in RAW264.7 macrophages and in the intestines of a mouse model. The hybrid peptide exhibited increased anti-inflammatory activity compared to the parental peptides. C-L plays a role in protecting intestinal tissue from damage, LPS-induced weight loss, and leukocyte infiltration. In addition, C-L reduces the expression levels of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), IL-1ß, and interferon-gamma (IFN-γ), as well as reduces cell apoptosis. It also reduced mucosal barrier damage caused by LPS. The anti-inflammatory effects of the hybrid peptide were mainly attributed to its LPS-neutralizing activity and antagonizing the activation of LPS-induced Toll-like receptor 4-myeloid differentiation factor 2 (TLR4/MD2). The peptide also affected the TLR4-(nuclear factor κB) signaling pathway, modulating the inflammatory response upon LPS stimulation. Collectively, these findings suggest that the newly designed peptide, C-L, could be developed into a novel anti-inflammatory agent for animals or humans.
Asunto(s)
Antiinflamatorios/farmacología , Inflamación/tratamiento farmacológico , Mucosa Intestinal/efectos de los fármacos , Péptidos/farmacología , Animales , Péptidos Catiónicos Antimicrobianos/farmacología , Línea Celular , Inflamación/inducido químicamente , Inflamación/metabolismo , Interleucina-6/metabolismo , Mucosa Intestinal/metabolismo , Lipopolisacáridos/farmacología , Masculino , Ratones , Ratones Endogámicos C57BL , Membrana Mucosa/efectos de los fármacos , Membrana Mucosa/metabolismo , FN-kappa B/metabolismo , Células RAW 264.7 , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 4/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
CLP is a novel hybrid peptide derived from CM4, LL37 and TP5, with significantly reduced hemolytic activity and increased antibacterial activity than parental antimicrobial peptides. To avoid host toxicity and obtain high-level bio-production of CLP, we established a His-tagged SUMO fusion expression system in Escherichia coli. The fusion protein can be purified using a Nickel column, cleaved by TEV protease, and further purified in flow-through of the Nickel column. As a result, the recombinant CLP with a yield of 27.56 mg/L and a purity of 93.6% was obtained. The purified CLP exhibits potent antimicrobial activity against gram+ and gram- bacteria. Furthermore, the result of propidium iodide staining and scanning electron microscopy (SEM) showed that CLP can induce the membrane permeabilization and cell death of Enterotoxigenic Escherichia coli (ETEC) K88. The analysis of thermal stability results showed that the antibacterial activity of CLP decreases slightly below 70 °C for 30 min. However, when the temperature was above 70 °C, the antibacterial activity was significantly decreased. In addition, the antibacterial activity of CLP was stable in the pH range from 4.0 to 9.0; however, when pH was below 4.0 and over 9.0, the activity of CLP decreased significantly. In the presence of various proteases, such as pepsin, papain, trypsin and proteinase K, the antibacterial activity of CLP remained above 46.2%. In summary, this study not only provides an effective strategy for high-level production of antimicrobial peptides and evaluates the interference factors that affect the biological activity of hybrid peptide CLP, but also paves the way for further exploration of the treatment of multidrug-resistant bacterial infections.
Asunto(s)
Antibacterianos/química , Péptidos Antimicrobianos/química , Péptidos/química , Proteínas Recombinantes de Fusión/genética , Antibacterianos/biosíntesis , Antibacterianos/farmacología , Péptidos Catiónicos Antimicrobianos/química , Péptidos Catiónicos Antimicrobianos/genética , Péptidos Antimicrobianos/biosíntesis , Péptidos Antimicrobianos/genética , Péptidos Antimicrobianos/farmacología , Bacterias/efectos de los fármacos , Bacterias/patogenicidad , Catelicidinas/química , Catelicidinas/genética , Escherichia coli/genética , Hemólisis/efectos de los fármacos , Humanos , Péptidos/genética , Péptidos/farmacología , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/farmacologíaRESUMEN
EF-1 is a novel peptide derived from two bacteriocins, plantaricin E and plantaricin F. It has a strong antibacterial activity against Escherichia coli and with negligible hemolytic effect on red blood cells. However, the chemical synthesis of EF-1 is limited by its high cost. In this study, we established a heterologous expression of EF-1 in Pichia pastoris. The transgenic strain successfully expressed hybrid EF-1 peptide, which had a molecular weight of ~5 kDa as expected. The recombinant EF-1 was purified by Ni2+ affinity chromatography and reversed-phase high performance liquid chromatography (RP-HPLC), which achieved a yield of 32.65 mg/L with a purity of 94.9%. The purified EF-1 exhibited strong antimicrobial and bactericidal activities against both Gram-positive and -negative bacteria. Furthermore, propidium iodide staining and scanning electron microscopy revealed that EF-1 can directly induce cell membrane permeabilization of E. coli. Therefore, the hybrid EF-1 not only preserves the individual properties of the parent peptides, but also acquires the ability to disrupt Gram-negative bacterial membrane. Meanwhile, such an expression system can reduce both the time and cost for large-scale peptide production, which ensures its potential application at the industrial level.
Asunto(s)
Antiinfecciosos/farmacología , Bacterias/efectos de los fármacos , Expresión Génica , Factor 1 de Elongación Peptídica/genética , Factor 1 de Elongación Peptídica/farmacología , Péptidos/genética , Péptidos/farmacología , Pichia/genética , Proteínas Recombinantes , Antiinfecciosos/aislamiento & purificación , Bacterias/metabolismo , Bacterias/ultraestructura , Permeabilidad de la Membrana Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Pruebas de Sensibilidad Microbiana , Factor 1 de Elongación Peptídica/química , Factor 1 de Elongación Peptídica/aislamiento & purificación , Péptidos/aislamiento & purificaciónRESUMEN
AIM: In this study, a questionnaire survey was conducted to find the relationship between preeclampsia (PE) and incontinentia pigmenti (IP). METHODS: Using a questionnaire survey of 147 women whose children were diagnosed with IP, this study first investigated their clinical manifestations and complications during pregnancy. The manifestations included high blood pressure, proteinuria and edema after 20 weeks of gestation. Women with and without IP were separated into two groups, then analyzed accordingly. RESULTS: There were 45 mothers with IP in the case group and 102 mothers without IP in the control group. IP mothers who were pregnant with an IP fetus were at higher risk for hypertension, proteinuria, and edema during pregnancy as compared with non-IP mothers that carried an IP fetus. Out of these 147 mothers, 8 mothers with IP and 6 mothers without IP presented with new-onset hypertension during pregnancy (P = 0.024)ï¼7 mothers with IP and 4 mothers without IP presented with new-onset proteinuria during pregnancy (P = 0.013)ï¼and 21 IP mothers and 27 non-IP mothers presented with edema during pregnancy (P = 0.016). Although no statistical difference was observed, mothers in the case group were more likely to develop the above three symptoms concurrently (6.7% vs 2.0%; P = 0.168), and were more likely to be diagnosed with PE (8.9% vs 3.9%; P = 0.249). CONCLUSION: Our study revealed that the simultaneous occurrence of IP in the mother and fetus increased the likelihood of clinical manifestations associated with PE during pregnancy.
Asunto(s)
Hipertensión Inducida en el Embarazo/genética , Incontinencia Pigmentaria/genética , Enfermedades del Recién Nacido/genética , Preeclampsia/genética , Adulto , Femenino , Humanos , Recién Nacido , Embarazo , Factores de Riesgo , Encuestas y CuestionariosRESUMEN
Intestinal inflammation is an inflammatory disease resulting from immune dysregulation in the gut. It can increase the risk of enteric cancer, which is a common malignancy globally. As a new class of anti-inflammatory agents, native peptides have potential for use in the treatment of several intestinal inflammation conditions; however, their potential cytotoxicity and poor anti-inflammatory activity and stability have prevented their development. Hybridization has been proposed to overcome this problem. Thus, in this study, we designed a hybrid peptide (LL-37-TP5, LTP) by combing the active centre of LL-37 (13-36) with TP5. The half-life and cytotoxicity were tested in vitro, and the hybrid peptide showed a longer half-life and lower cytotoxicity than its parental peptides. We also detected the anti-inflammatory effects and mechanisms of LTP on Lipopolysaccharide (LPS)-induced intestinal inflammation in murine model. The results showed that LTP effectively prevented LPS-induced weight loss, impairment of intestinal tissues, leukocyte infiltration, and histological evidence of inflammation. Additionally, LTP decreased the levels of tumour necrosis factor-alpha, interferon-gamma, and interleukin-6; increased the expression of zonula occludens-1 and occludin; and reduced permeability in the jejunum of LPS-treated mice. Notably, LTP appeared to be more potent than the parental peptides LL-37 and TP5. The anti-inflammatory effects of LTP may be associated with the neutralization of LPS, inhibition of oxidative stress, and inhibition of the NF-κB signalling pathway. The findings of this study suggest that LTP might be an effective therapeutic agent for treating intestinal inflammation.
Asunto(s)
Antioxidantes/uso terapéutico , Inflamación/inducido químicamente , Inflamación/tratamiento farmacológico , Mucosa Intestinal/metabolismo , Lipopolisacáridos/toxicidad , Animales , Interleucina-6/metabolismo , Mucosa Intestinal/efectos de los fármacos , Ratones , FN-kappa B/metabolismo , Ocludina/metabolismo , Células RAW 264.7 , Transducción de Señal/efectos de los fármacos , Proteína de la Zonula Occludens-1/metabolismoRESUMEN
Immunity is a defensive response that fights disease by identifying and destroying harmful substances or microbiological toxins. Several factors, including work-related stress, pollution, and immunosuppressive agents, contribute to low immunity and poor health. Native peptides, a new class of immunoregulatory agents, have the potential for treating immunodeficiencies, malignancies, and infections. However, the potential cytotoxicity and low immunoregulatory activity and stability of native peptides have prevented their development. Therefore, we designed three hybrid peptides (LTAa, LTAb, and LTAc) by combining a characteristic fragment of LL-37 with an active Tα1 center that included Tα1 (17-24), Tα1 (20-25), and Tα1 (20-27). The best hybrid peptide (LTAa), according to molecule docking and in vitro experiments, had improved immunoregulatory activity and stability with minimal cytotoxicity. We investigated the immunoregulatory effects and mechanisms of LTAa using a cyclophosphamide-immunosuppressed murine model. LTAa effectively reversed immunosuppression by enhancing immune organ development, activating peritoneal macrophage phagocytosis, regulating T lymphocyte subsets, and increasing cytokine (tumor necrosis factor-alpha, interleukin-6, and interleukin-1ß) and immunoglobulin (IgA, IgG, and IgM) contents. The immunomodulatory effects of LTAa may be associated with binding to the TLR4/MD-2 complex and activation of the NF-κB signaling pathway. Therefore, LTAa could be an effective therapeutic agent for improving immune function.
Asunto(s)
Factores Inmunológicos/síntesis química , Factores Inmunológicos/farmacología , Péptidos/síntesis química , Péptidos/farmacología , Animales , Western Blotting , Supervivencia Celular/efectos de los fármacos , Dicroismo Circular , Ensayo de Inmunoadsorción Enzimática , Femenino , Citometría de Flujo , Factores Inmunológicos/química , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/metabolismo , Ratones , Ratones Endogámicos BALB C , Péptidos/química , Células RAW 264.7RESUMEN
The hybrid peptide cecropin A (1â»8)â»LL37 (17â»30) (Câ»L), derived from the sequence of cecropin A (C) and LL-37 (L), showed significantly increased antibacterial activity and minimized hemolytic activity than C and L alone. To obtain high-level production of Câ»L, the deoxyribonucleic acid sequence encoding Câ»L with preferred codons was cloned into pET-SUMO to construct a fusion expression vector, and overexpressed in Escherichia coli (E. coli) BL21 (DE3). The maximum fusion protein (92% purity) was obtained with the yield of 89.14 mg/L fermentation culture after purification with Ni-NTA Sepharose column. The hybrid Câ»L was cleaved from the fusion protein by SUMO-protease, and 17.54 mg/L pure active Câ»L was obtained. Furthermore, the purified Câ»L showed identical antibacterial and hemolytic activity to synthesized Câ»L. Stability analysis results exhibited that the activity of Câ»L changed little below 80 °C for 20 min, but when the temperature exceeded 80 °C, a significant decrease was observed. Varying the pH from 5.0 to 10.0 did not appear to influence the activity of Câ»L, however, pH below 4.0 decreased the antibacterial activity of Câ»L rapidly. Under the challenge of several proteases (pepsin, trypsin, and proteinase K), the functional activity of Câ»L was maintained over 50%. In summary, this study not only supplied an effective approach for high-level production of hybrid peptide Câ»L, but paved the way for its further exploration in controlling infectious diseases of farm animals or even humans.
Asunto(s)
Antibacterianos/química , Péptidos Catiónicos Antimicrobianos/química , Proteínas Recombinantes de Fusión/química , Antibacterianos/aislamiento & purificación , Antibacterianos/farmacología , Diseño de Fármacos , Escherichia coli , Hemólisis , Humanos , Concentración de Iones de Hidrógeno , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/aislamiento & purificación , Proteínas Recombinantes de Fusión/farmacología , Staphylococcus aureus/efectos de los fármacos , Temperatura , CatelicidinasRESUMEN
BACKGROUND: Xylanase degrades xylan into monomers of various sizes by catalyzing the endohydrolysis of the 1,4-ß-D-xylosidic linkage randomly, possessing potential in wide industrial applications. Most of xylanases are susceptible to be inactive when suffering high temperature and high alkaline process. Therefore, it is necessary to develop a high amount of effective thermoalkaliphilic xylanases. This study aims to enhance thermoalkaliphilic xylanase production in Pichia pastoris through fermentation parameters optimization and novel efficient fed-batch strategy in high cell-density fermentation. RESULTS: Recombinant xylanase activity increased 12.2%, 7.4%, 12.0% and 9.9% by supplementing the Pichia pastoris culture with 20 g/L wheat bran, 5 mg/L L-histidine, 10 mg/L L-tryptophan and 10 mg/L L-methionine in shake flasks, respectively. Investigation of nutritional fermentation parameters, non-nutritional fermentation parameters and feeding strategies in 1 L bioreactor and 1 L shake flask revealed that glycerol and methanol feeding strategies were the critical factors for high cell density and xylanase activity. In 50 L bioreactor, a novel glycerol feeding strategy and a four-stage methanol feeding strategy with a stepwise increase in feeding rate were developed to enhance recombinant xylanase production. In the initial 72 h of methanol induction, the linear dependence of xylanase activity on methanol intake was observed (R2 = 0.9726). The maximum xylanase activity was predicted to be 591.2 U/mL, while the actual maximum xylanase activity was 560.7 U/mL, which was 7.05 times of that in shake flask. Recombinant xylanase retained 82.5% of its initial activity after pre-incubation at 80 °C for 50 min (pH 8.0), and it exhibited excellent stability in the broad temperature (60-80 °C) and pH (pH 8.0-11.0) ranges. CONCLUSIONS: Efficient glycerol and methanol fed-batch strategies resulting in desired cell density and xylanase activity should be applied in other P. pastoris fermentation for other recombinant proteins production. Recombinant xylanases with high pH- and thermal-stability showed potential in various industrial applications.
Asunto(s)
Actinobacteria/genética , Reactores Biológicos/microbiología , Endo-1,4-beta Xilanasas/biosíntesis , Fermentación/fisiología , Glicerol/metabolismo , Metanol/metabolismo , Pichia/fisiología , Actinobacteria/enzimología , Técnicas de Cultivo Celular por Lotes , Recuento de Células , Proliferación Celular/fisiología , Medios de Cultivo/metabolismo , Endo-1,4-beta Xilanasas/genética , Endo-1,4-beta Xilanasas/aislamiento & purificación , Activación Enzimática , Mejoramiento Genético/métodos , Pichia/clasificación , Pichia/citología , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Especificidad de la EspecieRESUMEN
Hybridizing different antimicrobial peptides (AMPs) is a particularly successful approach to obtain novel AMPs with increased antimicrobial activity but minimized cytotoxicity. The hybrid peptide cecropin A (1-8)-LL37 (17-30) (C-L) combining the hydrophobic N-terminal fragment of cecropin A (C) with the core antimicrobial fragment of LL37 (L) was designed and synthesized. C-L showed higher antibacterial activity against all indicator strains than C and L, and no hemolytic activity to sheep erythrocytes was observed. C-L kills bacterial cells and causes disruption of surface structure, as determined by scanning electron microscopy. Synergistic effects were observed in the combination of C-L with several antibiotics (chloramphenicol, thiamphenicol, or neomycin sulfate) against Escherichia coli and Staphylococcus aureus.
Asunto(s)
Antibacterianos/farmacología , Péptidos Catiónicos Antimicrobianos/farmacología , Animales , Antibacterianos/efectos adversos , Péptidos Catiónicos Antimicrobianos/efectos adversos , Cloranfenicol/efectos adversos , Cloranfenicol/farmacología , Eritrocitos/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Hemólisis/efectos de los fármacos , Microscopía Electrónica de Rastreo , Neomicina/efectos adversos , Neomicina/farmacología , Ovinos , Staphylococcus aureus/efectos de los fármacos , Tianfenicol/efectos adversos , Tianfenicol/farmacologíaRESUMEN
All life generates deoxyribonucleotides, the building blocks of DNA, via ribonucleotide reductases (RNRs). The complexity of this reaction suggests it did not evolve until well after the advent of templated protein synthesis, which in turn suggests DNA evolved later than both RNA and templated protein synthesis. However, deoxyribonucleotides may have first been synthesised via an alternative, chemically simpler route--the reversal of the deoxyriboaldolase (DERA) step in deoxyribonucleotide salvage. In light of recent work demonstrating that this reaction can drive synthesis of deoxyribonucleosides, we consider what pressures early adoption of this pathway would have placed on cell metabolism. This in turn provides a rationale for the replacement of DERA-dependent DNA production by RNR-dependent production.
Asunto(s)
Aldehído-Liasas/química , Evolución Biológica , ADN/química , Origen de la Vida , Ribonucleótido Reductasas/química , Aldehído-Liasas/metabolismo , ADN/metabolismo , Replicación del ADN , Desoxirribonucleótidos/química , Desoxirribonucleótidos/metabolismo , Eucariontes/química , Eucariontes/metabolismo , Células Procariotas/química , Células Procariotas/metabolismo , Ribonucleótido Reductasas/metabolismo , Factores de TiempoRESUMEN
BACKGROUND: Variants in the emerin gene (EMD) were implicated in X-linked recessive Emery-Dreifuss muscular dystrophy (EDMD), characterized by early-onset contractures of tendons, progressive muscular weakness and cardiomyopathy. To date, 223 mutations have been reported in EMD gene and the majority of them caused a predominant skeletal muscular phenotype. In this study, we identified a novel deletion mutation in EMD exon 1, which results in almost a complete loss of emerin protein in a large Chinese family. However, the patients suffered severe dilated cardiomyopathy (DCM) but very mild skeletal muscle disorder. CASE PRESENTATION: Whole exome sequencing (WES) and linkage analysis were performed to identify the underlying mutation in a Chinese DCM family spanning five generations. A missense variation in the GPR50 gene was found co-segregated with the disease phenotype, whereas no functional alteration was detected in the variant GPR50 protein. When analyzing the failure sequences in the exome sequencing data, a novel deletion mutation (c.26_39delATACCGAGCTGACC) in EMD exon 1, was identified in this family. Different from the typical clinical features caused by most reported EMD mutations, patients in our study presented very mild skeletal muscle degeneration that had not been diagnosed until the mutation was found. CONCLUSION: We described a family with rare clinical presentations caused by a novel EMD deletion mutation. Our findings broaden the heterogeneous spectrum of phenotypes attributed to EMD mutations and provide new insight to explain the genotype-phenotype correlations between EMD mutations and EDMD symptoms.
Asunto(s)
Cardiomiopatía Dilatada/genética , Proteínas de la Membrana/genética , Distrofia Muscular de Emery-Dreifuss/genética , Proteínas del Tejido Nervioso/genética , Proteínas Nucleares/genética , Receptores Acoplados a Proteínas G/genética , Eliminación de Secuencia , China , Exoma , Exones , Femenino , Estudios de Asociación Genética , Heterogeneidad Genética , Ligamiento Genético , Variación Genética , Genotipo , Humanos , Masculino , Distrofia Muscular de Emery-Dreifuss/patología , Linaje , Fenotipo , Análisis de Secuencia de ADNRESUMEN
OBJECTIVES: Marked interpatient variability exists in the blood pressure response to carvedilol, a nonselective ß-blocker. Here we evaluated the influence of 4 common polymorphisms in genes of the ß-adrenergic receptor on the antihypertensive efficacy of carvedilol in patients in a double-blinded monotherapy study. METHODS: Eighty-seven subjects with uncomplicated essential hypertensive (49% men; age = 52.2 ± 11.1 years) from Jilin province of China were enrolled in the study, and 5 of them discontinued the treatment due to adverse effects. Both systolic and diastolic blood pressures (DBPs) were measured before and after 7 days of treatment with carvedilol (10 mg/d). Genotypes of the ß1-adrenergic receptor (ADRB1 Ser49Gly and Arg389Gly) and ß2-adrenergic receptor (ADRB2 Gly16Arg and Glu27Gln) were determined by polymerase chain reaction with restriction fragment length polymorphism. RESULTS: Patients homozygous for ADRB1 Arg389 had an approximately 4-fold greater reduction in DBPs than those homozygous for ADRB1 Gly389 (10.61 vs. 2.62 mm Hg, P = 0.013). The ADRB1 haplotype was also a significant predictor of response, as patients with the Gly49Arg389/Ser49Arg389 haplotype pair had a 5.7-fold greater reduction in DBPs than those homozygous for the Ser49Gly389 haplotype (16.11 vs. 2.83 mm Hg, P = 0.0055). An association was not found between ADRB2 polymorphism and carvedilol responsiveness in antihypertensive therapy. CONCLUSIONS: This study provides the first evidence to support that ADRB1 polymorphisms play an important role in the DBPs response to carvedilol treatment in patients with essential hypertension.
Asunto(s)
Antihipertensivos/uso terapéutico , Carbazoles/uso terapéutico , Hipertensión/tratamiento farmacológico , Polimorfismo de Nucleótido Simple , Propanolaminas/uso terapéutico , Receptores Adrenérgicos beta 1/genética , Antihipertensivos/administración & dosificación , Antihipertensivos/efectos adversos , Presión Sanguínea/efectos de los fármacos , Carbazoles/administración & dosificación , Carbazoles/efectos adversos , Carvedilol , China , Hipertensión Esencial , Femenino , Haplotipos , Heterocigoto , Homocigoto , Humanos , Hipertensión/genética , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Propanolaminas/administración & dosificación , Propanolaminas/efectos adversosRESUMEN
Recent evidence suggests that mulberry leaves have good antioxidant activity. However, what the antioxidant ingredient is and how the ingredient works are still not well understood. In this study, we enzymatically hydrolyze mulberry leaf proteins (MLPs) using neutral protease and find that the mulberry leaf protein hydrolysates (MLPHs) have stronger antioxidant activity compared to MLPs. We separate the core antioxidant components in MLPHs by ion-exchange columns and molecular sieves and identify 798 antioxidant peptides by LC-MS/MS. Through bioinformatics analysis and biochemical assays, we screen two previously unreported peptides, P6 and P7, with excellent antioxidant activities. P6 and P7 not only significantly reduce ROS in cells but also improve the activities of the antioxidant enzymes SOD and CAT. In addition, both peptides are found to exert protective effects against H2O2-induced chromatin damage and cell apoptosis. Collectively, these results provide support for the application of mulberry leaf peptides as antioxidants in the medical, food and livestock industries.
RESUMEN
Uracil-thymine dehydrogenase (UTDH), which catalyzes the irreversible oxidation of uracil to barbituric acid in oxidative pyrimidine metabolism, was purified from Rhodococcus erythropolis JCM 3132. The finding of unusual stabilizing conditions (pH 11, in the presence of NADP+ or NADPH) enabled the enzyme purification. The purified enzyme was a heteromer consisting of three different subunits. The enzyme catalyzed oxidation of uracil to barbituric acid with artificial electron acceptors such as methylene blue, phenazine methosulfate, benzoquinone, and α-naphthoquinone; however, NAD+, NADP+, flavin adenine dinucleotide, and flavin mononucleotide did not serve as electron acceptors. The enzyme acted not only on uracil and thymine but also on 5-halogen-substituted uracil and hydroxypyrimidine (pyrimidone), while dihydropyrimidine, which is an intermediate in reductive pyrimidine metabolism, and purine did not serve as substrates. The activity of UTDH was enhanced by cerium ions, and this activation was observed with all combinations of substrates and electron acceptors.
Asunto(s)
Oxidación-Reducción , Pirimidinas , Rhodococcus , Uracilo , Uracilo/metabolismo , Uracilo/química , Pirimidinas/metabolismo , Rhodococcus/enzimología , NADP/metabolismo , Azul de Metileno/metabolismo , Azul de Metileno/química , Barbitúricos/metabolismo , Barbitúricos/química , Benzoquinonas/metabolismo , Benzoquinonas/química , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/química , Concentración de Iones de Hidrógeno , Timina/metabolismo , Timina/química , Especificidad por Sustrato , Metosulfato de Metilfenazonio/metabolismo , Metosulfato de Metilfenazonio/químicaRESUMEN
In this study, the yield of exopolysaccharide (EPS) from Lactobacillus plantarum R301 was optimized using a single-factor experiment and response surface methodology (RSM). After optimization, the EPS yield was increased with a fold-change of 0.85. The significant factors affecting EPS production, as determined through a Plackett-Burman design and Central Composite Design (CCD), were MgSO4 concentration, initial pH, and inoculation size. The maximum yield was 97.85 mg/mL under the condition of 0.01% MgSO4, an initial pH 7.4, and 6.4% of the inoculation size. In addition, the EPS exhibited strong antioxidant activity, as demonstrated by its ability to scavenge DPPH, ABTS, and hydroxyl radicals. The scavenging rate was up to 100% at concentrations of 4 mg/mL, 1 mg/mL, and 2 mg/mL, respectively. Moreover, the EPS also exhibited reducing power, which was about 30% that of ascorbic acid when both tended to be stable with the increased concentration. These results suggest that L. plantarum R301 EPS possesses different antioxidant mechanisms and warrants further investigation. In addition to its antioxidant activity, the EPS also demonstrated good anti-inflammatory activity by inhibiting the inflammation induced by lipopolysaccharide (LPS) in RAW 264.7 cells, which could decrease nitric oxide (NO) production and expression of the proinflammatory cytokine Il-6. These findings suggest that L. plantarum R301 EPS could be used as a potential multifunctional food additive in the food industry.
RESUMEN
The current study aimed to investigate the effects of Clostridium butyiricum on growth performance, intestinal morphology, serum biochemical response, and immunity in broiler chickens. A total of 330 commercial one-day-old, mixed-sex Ross 308 broilers were randomly divided into five treatment groups with six replicates per group. The broilers were fed the basal diet (CON), the basal diet with 150 mg/kg of aureomycin (AM), the basal diet with C. butyricum at 2 × 108 CFU/kg (CBL), the basal diet with C. butyricum at 4 × 108 CFU/kg (CBM), and the basal diet with C. butyricum at 8 × 108 CFU/kg (CBH). Results showed that the final body weight (BW) (p < 0.01; p < 0.05), ADG from day 22 to 39 (p < 0.05), and ADG from day 1 to 39 (p < 0.01; p < 0.05) were improved in a linear and quadratic response with the inclusion of C. butyricum. There were no differences in feed conversion rate (FCR) among all groups (p > 0.05). Supplementation with C. butyricum quadratically reduced the crypt depth at day 21 (p < 0.01), linearly improved the villus height in the jejunum at day 39 (p < 0.001), and linearly and quadratically increased the villus height to crypt depth (V/C) ratio in the jejunum at day 21 (p < 0.01) and day 39 (p < 0.01; p < 0.001). Dietary C. butyricum affected the thymus index at day 21 and day 39 (linear, p < 0.01), and the bursa of Fabricius index at day 39 (quadratic, p < 0.05). Compared to the AM group, the serum urea contents were decreased (p < 0.05) but the IgG contents were increased in the CBL and CBH groups at day 21 (p < 0.01); in addition, serum albumin (ALB) concentrations in all the C. butyricum-supplemented groups (p < 0.01) and IgG concentrations in the CBM group were augmented at day 39 (p < 0.05). In conclusion, dietary C. butyricum could enhance growth performance by improving jejunal morphology and stimulating immunity organ development in broilers, and could be an alternative to antibiotics in poultry feeds.