RESUMEN
Insertion/deletion polymorphism (InDel) genetic markers refer to insertion or deletion of DNA fragments into genomic DNA, which have advantages in the identification of degraded samples. In this study, we independently screened 66 highly polymorphic InDel markers from the dbSNP database to establish a multiplex PCR system for forensic DNA identification using next-generation sequencing system (66-plex InDels). We assessed the population genetic data among 251 Chinese Han population using this system and evaluated their potential forensic application. The results showed that all 66 InDel loci conformed to the Hardy-Weinberg equilibrium (P>0.000 758), and all the pairwise InDel loci were in linkage equilibrium after Bonferroni correction. The mean observed heterozygosity (Ho) was 0.482, the mean expected heterozygosity (He) was 0.483,the mean discrimination power (DP) was 0.612, the mean polymorphism information content (PIC) was 0.365, the total discrimination power (TDP) reached 0.999 999 999 999 999 999 999 999 999 428 18. The cumulative power of exclusion for 66 InDel loci was 0.999 739 in duo cases (CPEduo) and was 0.999 999 999 417 in trios cases (CPEtrio). The results show that the 66 InDel loci have high genetic polymorphisms in the Chinese Han population and can be used independently for forensic DNA identification and paternity testing.
Asunto(s)
Mutación INDEL , Polimorfismo Genético , China , ADN/genética , Frecuencia de los Genes , Sitios Genéticos , Genética de Población , Humanos , Repeticiones de MicrosatéliteRESUMEN
To improve nicotinic acid (NA) yield and meet industrial application requirements of sodium alginate-polyvinyl alcohol (SA-PVA) immobilized cells of Pseudomonas putida mut-D3 harboring nitrilase, inorganic materials were added to the SA-PVA immobilized cells to improve mechanical strength and mass transfer performance. The concentrations of inorganic materials were optimized to be 2.0% silica and 0.6% CaCO3. The optimal pH and temperature for SA-PVA immobilized cells and composite immobilized cells were both 8.0 and 45 °C, respectively. The half-lives of composite immobilized cells were 271.48, 150.92, 92.92 and 33.12 h, which were 1.40-, 1.35-, 1.22- and 1.63-fold compared to SA-PVA immobilized cells, respectively. The storage stability of the composite immobilized cells was slightly increased. The composite immobilized cells could convert 14 batches of 3-cyanopyridine with feeding concentration of 250 mM and accumulate 418 g ·L-1 nicotinic acid, while the SA-PVA immobilized cells accumulated 346 g L-1 nicotinic acid.
Asunto(s)
Alginatos/química , Aminohidrolasas/química , Alcohol Polivinílico/química , Pseudomonas putida/enzimología , Biocatálisis , Carbonato de Calcio , Células Inmovilizadas , Ácidos Hexurónicos , Hidroliasas , Concentración de Iones de Hidrógeno , Compuestos Inorgánicos , Microscopía Electrónica de Transmisión , Niacina/química , Piridinas/química , Dióxido de Silicio/química , TemperaturaRESUMEN
BACKGROUND: First flower node (FFN) is an important trait for evaluating fruit earliness in pepper (Capsicum annuum L.). The trait is controlled by quantitative trait loci (QTL); however, studies have been limited on QTL mapping and genes contributing to the trait. RESULTS: In this study, we developed a high density genetic map using specific-locus amplified fragment sequencing (SLAF-seq), a high-throughput strategy for de novo single nucleotide polymorphism discovery, based on 146 recombinant inbred lines (RILs) derived from an intraspecific cross between PM702 and FS871. The map contained 9328 SLAF markers on 12 linkage groups (LGs), and spanned a total genetic distance of 2009.69 centimorgan (cM) with an average distance of 0.22 cM. The sequencing depth for the map was 72.39-fold in the male parent, 57.04-fold in the female parent, and 15.65-fold in offspring. Using the genetic map, two major QTLs, named Ffn2.1 and Ffn2.2, identified on LG02 were strongly associated with FFN, with a phenotypic variance explanation of 28.62 and 19.56%, respectively. On the basis of the current annotation of C. annuum cv. Criollo de Morelos (CM334), 59 candidate genes were found within the Ffn2.1 and Ffn2.2 region, but only 3 of 59 genes were differentially expressed according to the RNA-seq results. Eventually we identified one gene associated with the FFN based on the function through GO, KEGG, and Swiss-prot analysis. CONCLUSIONS: Our research showed that the construction of high-density genetic map using SLAF-seq is a valuable tool for fine QTL mapping. The map we constructed is by far the most saturated complete genetic map of pepper, and using it we conducted fine QTL mapping for the important trait, FFN. QTLs and candidate genes obtained in this study lay a good foundation for the further research on FFN-related genes and other genetic applications in pepper.
Asunto(s)
Capsicum/genética , Mapeo Cromosómico/métodos , Cromosomas de las Plantas , Flores/genética , Sitios de Carácter Cuantitativo , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Ligamiento Genético , Genotipo , Secuenciación de Nucleótidos de Alto Rendimiento , Polimorfismo de Nucleótido SimpleRESUMEN
3-Hydroxypropionic acid (3-HP) is an important platform chemical in organic synthesis. Traditionally, 3-HP was produced by chemical methods and fermentation process. In this work, a novel enzymatic method was developed for green synthesis of 3-HP. A yeast strain harboring nitrile-hydrolyzing enzyme was newly isolated from environmental samples using 3-hydroxypropionitrile (3-HPN) as the sole nitrogen source. It was identified to be Meyerozyma guilliermondii CGMCC12935 by sequencing of the 18S ribosomal DNA and internal transcribed spacer, together with analysis of the morphology characteristics. The catalytic properties of M. guilliermondii CGMCC12935 resting cells were determined, and the optimum activity was achieved at 55 °C and pH 7.5. The enzyme showed broad substrate specificity towards nitriles, especially 3-HPN, aminoacetonitrile and 3-cyanopyridine. The presence of Ag+, Pb2+ and excess substrate inhibited the enzyme activity, whereas 5% (v/v) ethyl acetate had a positive effect on the enzyme activity. M. guilliermondii CGMCC12935 resting cells by addition of 3% glucose could thoroughly hydrolyze 500 mM 3-HPN into 3-HP within 100 h and the maximal accumulative production of 3-HP reached 216.33 mM, which was over twofolds than the control group with no additional glucose. And this work would lay the foundation for biological production of 3-HP in industry.
Asunto(s)
Candida , Ácido Láctico/análogos & derivados , Nitrilos , PiridinasRESUMEN
Due to the wide sources of biomass raw materials and the lack of limits for the endogenous pollutants in biochar and their dosage, some biochar with high endogenous pollutants may be used for environmental remediation, which results in potential environmental risks. In this study, three biochars were prepared from the straws of Pennisetum sp. grown in clean, moderately polluted and highly polluted soils, respectively. The total endogenous copper (Cu) and cadmium (Cd), acid-soluble fraction, and persistent free radical (PFRs) distribution in biochars were investigated, and their biotoxicities were evaluated based on wheat root elongation inhibition rate and antioxidant enzyme activity. The results indicated that total Cu in Jiuniu biochar from the highly polluted soil and total Cd in Shuiquan biochar from the moderately-polluted soil were 3.73 and 4.43 times higher than that in Hongrang biochar from the clean soil, respectively. Moreover, acid-soluble Cu in Jiuniu biochar was 3.32 and 2.84 times higher than that in Shuiquan and Hongrang biochar, respectively, and acid-soluble Cd in Shuiquan and Jiuniu biochar was 7.95 and 5.11 times higher than that in Hongrang biochar, respectively. All three biochars had PFRs with adjacent oxygen atoms centered on carbon and followed the order of Hongrang>Jiuniu>Shuiquan. Three biochar leaching solutions significantly inhibited wheat root elongation but enhanced the enzyme activities of SOD, POD, and CAT for the wheat seedlings compared with that in the control. In particular, the highest inhibition rate (27.7%) was found in Jiuniu biochar. This study indicated that the interaction of endogenous heavy metals and PFRs in biochar exhibited significant biotoxicity to wheat seedlings. In the future, more attention should be paid to the potential environmental toxicity of endogenous pollutants from biochar to avoid new environmental pollution problems.
Asunto(s)
Contaminantes Ambientales , Pennisetum , Cadmio/toxicidad , Contaminación Ambiental , Antioxidantes , PlantonesRESUMEN
Aim: The underlying mechanisms by which circular RNAs (circRNAs) regulate non-small-cell lung cancer (NSCLC) progression remain elusive. This study investigated the role of circRNA circTTBK2 in NSCLC tumorigenesis. Materials & methods: Quantitative reverse transcriptase polymerase chain reaction analysis of circTTBK2 in NSCLC tissues and cell lines was performed. Cell proliferation, migration, invasion and tumorigenesis were confirmed in vitro and in vivo using CCK-8, EdU incorporation, Transwell assays and xenograft technique. The circTTBK2/miR-873-5p/TEAD1/DERL1 axis was verified by RNA immunoprecipitation, chromatin immunoprecipitation and luciferase reporter assays. Results: Overexpressed circTTBK2 in NSCLC tissues indicates poor prognosis of NSCLC patients. circTTBK2 harbors miR-873-5p, and miR-873-5p directly targets TEAD1. TEAD1 transcriptionally activates DERL1. Conclusion: This study revealed a novel machinery of circTTBK2/miR-873-5p/TEAD1/DERL1 for NSCLC tumorigenesis.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas , Neoplasias Pulmonares , MicroARNs , ARN Circular , Apoptosis/genética , Carcinogénesis/genética , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/patología , Línea Celular Tumoral , Proliferación Celular/genética , Proteínas de Unión al ADN/genética , Regulación Neoplásica de la Expresión Génica , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patología , Proteínas de la Membrana/genética , MicroARNs/genética , Proteínas Nucleares/genética , ARN Circular/genética , Factores de Transcripción de Dominio TEA/genéticaRESUMEN
Background: The aim of the study was to evaluate the efficacy of abdominopelvic lymphatic drainage area irradiation (APLN), instead of whole abdominal radiotherapy (WART), in the consolidative radiotherapy of advanced ovarian carcinoma patients. Methods: We conducted a retrospective analysis collecting 99 patients with locally advanced ovarian cancer treated by APLN with 45 - 50 Gy/25- 28 fractions/5-7#, instead of WART. We evaluated the clinical outcome of APLN. Five patients were selected for dosimetric verifications verses WART (30 Gy/20 fractions). The normal tissue complication probability (NTCP) was calculated for the two treatment methods. Results: The mean follow-up time was 64.10 months (5.5 - 113.2 months), after APLN consolidative radiotherapy, 1-, 3-, and 5-year overall survival (OS) was 87.9%, 81.3%, and 61.5%, median disease-free survival (DFS) was 40.8 months, 5-year local recurrence free survival (LRFS) was 75.9%, and 5-year distant metastasis free survival (DMFS) was 49.2%. One patient died due to intestinal perforation. Local recurrence in the area between WART and APLN was rare (3/99 patients). The number of surgical procedures < 2 was an independent risk factor for LRFS (P = 0.023). Dosimetric comparison showed that comparing with WART, APLN significantly reduced the organ at risk (OAR) dose: 25.37 ± 3.63 Gy (25%) for liver, 8.77 ± 5.03 Gy (25%) for kidney, 8.14 ± 1.51 Gy (25%) for small intestine, etc. NTCP was reduced by 0.04-1.04% for liver, kidney, and small intestine. Conclusion: For consolidative radiotherapy in locally advanced ovarian cancer, APLN (intensity-modulated radiotherapy 45 - 50 Gy/25 - 28 fractions) could be an alternative to WART, resulting in excellent LRFS and DFS, with acceptable toxicities, comparing with previous literature reports. Dosimetric analysis also showed the benefits of APLN in NTCP.
RESUMEN
Germline mutations in the RET proto-oncogene (RET gene) are well documented as the genetic causes of multiple endocrine neoplasia type 2A (MEN2A). We performed genetic analysis by direct RET gene mutation analysis in a Chinese MEN2A family and compared these results with biochemical screening tests and pathological examinations. Twenty-one exons and flanking introns of the RET gene were amplified using polymerase chain reaction (PCR). The PCR products were subjected to sequencing directly, or cloned into pGEM-T plasmids and sequenced. Restriction fragment length polymorphism (RFLP) was employed to confirm the mutation on the RET sequence. A novel heterozygous mutation of a 3-bp (GAC) deletion at codon 631 (D631del) of exon 11, resulting in the deletion of an aspartic acid at the locus, was identified in four MEN2A patients and one phenotypically normal family member. The average clinical onset-age of four MEN2A patients was 33.7 years, no cervical lymph node metastasis was found in MEN2A patients with medullary thyroid carcinoma in the family. The study indicated that the novel heterozygous deletion mutation at D631 of RET gene was co-segregated with MEN2A phenotype and promoted the development of MEN2A. This report is the first description of the D631del mutation in the family with MEN2A.
Asunto(s)
Predisposición Genética a la Enfermedad , Neoplasia Endocrina Múltiple Tipo 2a/genética , Linaje , Proteínas Proto-Oncogénicas c-ret/genética , Eliminación de Secuencia , Adulto , Niño , China , Familia , Femenino , Ligamiento Genético , Humanos , Masculino , Proto-Oncogenes MasRESUMEN
OBJECTIVE: To investigate the characteristics of post-transplantation diabetes mellitus and analyze its risk factors. METHODS: Extensive survey was carried out to understand the characteristics of posttransplantation diabetes mellitus in patients who received kidney grafting from February 1984 to December 2006. RESULTS: Three hundred forty-four post-transplantation diabetes mellitus patients from 1872 ones after kidney grafting were found from February 1984 to December 2006. The prevalence of new onset posttransplant diabetes mellitus and impaired fasting glucose in kidney allograft recipients were 18.4% and 12.7% respectively, being significantly higher than that in general population and other inpatients. The options of immunosuppressants were significantly associated with the prevalence of post-transplantation diabetes mellitus. By multivariate logistic regression analysis, the baseline characteristics of the post transplantation diabetes mellitus patients were significantly associated with increased age (OR: 1.309, P = 0.049), elevated level of the triglyceride ( OR: 1.311, P = 0.005), treatment with tacrolimus (FK506) (OR: 1.522, P = 0.008), and large dose of intravenous pulsed prednisolone (OR: 1.239, P = 0.011), as compared with patients without post-transplantation diabetes mellitus. Besides, the number of patients with at least one acute rejection episode was significantly greater in the post-transplantation diabetic patients. Mycophenolate mofetil (OR: 0.716, P = 0.028) and diltiazem (OR: 0.737, P = 0.015) were associated with lower risk of post-transplantation diabetes mellitus. CONCLUSIONS: High prevalence of abnormal glucose metabolism in renal allograft recipients during hospitalization was observed. Many risk factors contributed to the development of post transplantation diabetes mellitus.
Asunto(s)
Diabetes Mellitus/etiología , Trasplante de Riñón/efectos adversos , Complicaciones Posoperatorias/etiología , Adolescente , Adulto , Femenino , Humanos , Inmunosupresores/efectos adversos , Incidencia , Modelos Logísticos , Masculino , Persona de Mediana Edad , Análisis Multivariante , Factores de Riesgo , Adulto JovenRESUMEN
The role of protein phosphatase 2ACα (PP2ACα) in brain development is poorly understood. To understand the function of PP2ACα in neurogenesis, we inactivated Pp2acα gene in the central nervous system (CNS) of mice by Cre/LoxP system and generated the PP2ACα deficient mice (designated as the Pp2acα-/- mice). PP2ACα deletion results in DNA damage in neuroprogenitor cells (NPCs), which impairs memory formation and cortical neurogenesis. We first identify that PP2ACα can directly associate with Ataxia telangiectasia mutant kinase (ATM) and Ataxia telangiectasia/Rad3-related kinase (ATR) in neocortex and NPCs. Importantly, the P53 and hypermethylated in cancer 1 (HIC1) function complex, the newly found down-stream executor of the ATR/ATM cascade, will be translocated into nuclei and interact with homeodomain interacting protein kinase 2 (HIPK2) to respond to DNA damage. Notably, HICI plays a direct transcriptional regulatory role in HIPK2 gene expression. The interplay among P53, HIC1 and HIPK2 maintains DNA stability in neuroprogenitor cells. Taken together, our findings highlight a new role of PP2ACα in regulating early neurogenesis through maintaining DNA stability in neuroprogenitor cells. The P53/HIC/HIPK2 regulation loop, directly targeted by the ATR/ATM cascade, is involved in DNA repair in neuroprogenitor cells.
Asunto(s)
Encéfalo/crecimiento & desarrollo , Daño del ADN , Eliminación de Gen , Células-Madre Neurales/metabolismo , Proteína Fosfatasa 2C/deficiencia , Proteína Fosfatasa 2C/genética , Animales , Encéfalo/citología , Encéfalo/fisiología , Proteínas Portadoras/metabolismo , Proliferación Celular , Cognición , Histonas/metabolismo , Factores de Transcripción de Tipo Kruppel/metabolismo , Ratones , Microcefalia/enzimología , Microcefalia/genética , Neocórtex/metabolismo , Células-Madre Neurales/citología , Proteínas Serina-Treonina Quinasas/metabolismo , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
BACKGROUND: Crocin is the major chemical constituent of the Chinese herb saffron. A number of studies have indicated that crocin induces an antitumor effect by inhibiting proliferation and inducing the apoptosis of tumor cells. However, the effect of crocin on tumor angiogenesis remains unknown. METHODS: The effects of prolonged crocin exposure on breast cancer cell MDA-MB-231, human umbilical vein endothelial cells (HUVECs) and mice were examined. RESULTS: Crocin had a profound effect on the morphology and proliferation rate of MDA-MB-231 and HUVECs. Furthermore, crocin induced apoptosis and cell cycle arrest at the G2/M phase in MDA-MB-231 cells in a dose-dependent manner. This confirms that crocin induces the inhibition of HUVECs. Furthermore, the expression of CD34 in tumor tissues decreased after crocin treatment. CONCLUSIONS: Crocin has an anti-angiogenesis effect that may be correlated to the decreased expression of CD34. Crocin is likely to be involved in the regulation of molecules in the angiogenesis pathway.
RESUMEN
OBJECTIVE: To compare the different pregnancy outcomes of women with a reduced dichorionic triamniotic (DCTA) triplet managed with radiofrequency ablation (RFA) or potassium chloride (KCL). MATERIALS AND METHODS: This was a retrospective cohort study. We studied 30 women of DCTA triplets managed with RFA as well as 85 managed with KCL. We compared the mean neonatal birthweight, median gestational age and perinatal mortality of two groups. RESULTS: The mean neonatal birthweight of children in RFA group was 2572.4 g (SD, 407.0), vs 2899.3 g (SD, 554.9) in KCL group (P < 0.001). The rate of low birth weight infants was 23 (42.6%) vs. 16 (18.0%), respectively, (p < 0.005). However, there was no statistically significant difference in the median gestational age of delivery, premature birth before 32&37 weeks' gestation, neonatal brain injury or successful pregnancy between two groups. (We define the successful pregnancy as the condition that at least one child survives for a specific woman, while the failed one as no child survives.) CONCLUSION: What we took it for granted was that pregnancy outcomes in women with a reduced DCTA triplet managed with RFA was riskier than with KCL, however, we proved that it is not accurate. For women with a reduced DCTA triplet, managed with RFA is not much riskier than with KCL. What's more, most women have two children survived in RFA group, while in KCL group, only one child survives for most women. This result may change the management alternative for those women with DCTA triplet pregnancies who choose reduction, especially for women who desire to have two surviving and healthy fetuses.
Asunto(s)
Ablación por Catéter/métodos , Cloruro de Potasio/uso terapéutico , Reducción de Embarazo Multifetal/métodos , Embarazo Triple , Adulto , China/epidemiología , Femenino , Estudios de Seguimiento , Edad Gestacional , Humanos , Recién Nacido , Mortalidad Perinatal/tendencias , Embarazo , Resultado del Embarazo , Estudios RetrospectivosRESUMEN
Eight compounds were isolated from the seeds of Holarrhena antidysenterica Wall.ex A.DC. On the basis of physico-chemical properties and spectroscopic data, holarrhenanan (1) was identified as a new compound, compounds 2-3 were isolated from H. antidysenterica for the first time, and five known compounds were also obtained. Inhibitory effects of some compounds and extracts to the intestinal peristalsis were evaluated. Results showed that the extracts and compounds 4, 6 exhibited remarkable inhibitory effects with tension inhibition rate of 32.77, 32.77% and amplitude inhibition rate of 59.51, 55.98%, respectively on the vitro rabbit intestinal peristalsis.
Asunto(s)
Antidiarreicos/química , Holarrhena/química , Peristaltismo/efectos de los fármacos , Animales , Antidiarreicos/farmacología , Diarrea/tratamiento farmacológico , Intestinos/efectos de los fármacos , Espectroscopía de Resonancia Magnética , Estructura Molecular , Técnicas de Cultivo de Órganos , Extractos Vegetales/química , Extractos Vegetales/farmacología , Conejos , Semillas/químicaRESUMEN
The constitutively expression system for P. putida nitrilase was firstly constructed to improve the nicotinic acid production and reduce the production costs. High density culture strategy was employed to enhance the biomass and nitrilase production of recombinant strain. The total nitrilase activity reached up to 654â¯U·mL-1 without the induction. 541â¯g·L-1 nicotinic acid was accumulated via fed batch mode of substrate feeding through 290â¯min of conversion.
Asunto(s)
Aminohidrolasas , Biocatálisis , NiacinaRESUMEN
OBJECTIVE: To detect the mutations of RET proto-oncogene in two Chinese families with multiple endocrine neoplasia type 2A (MEN2A) and to study the clinical application of pentagastrin stimulation test in the diagnosis and follow-up of MEN2A. METHODS: In this study, there were 4 patients clinically diagnosed as MEN2A in the two pedigrees. PCR and direct gene sequencing of PCR products with an automated DNA sequencer were used to scan the entire 21 exons of RET proto-oncogene in the leukocyte DNA of 6 members of the two families. Blood calcitonin was measured and pentagastrin stimulation test carried out in all the 4 clinically diagnosed patients. RESULTS: Two missense mutations were detected in exon 11 of the RET proto-oncogene. One was C634R in the 2 patients from one MEN2A family and the other was C634Y in 2 patients from another family. The basal serum calcitonin was very high (400.5 - 13 510.7 ng/L, normal range 16.6 - 132.8 ng/L) in the MEN2A patients and after pentagastrin stimulation the serum calcitonin level was significantly higher (494.1 - 33 901.9 ng/L). CONCLUSIONS: Two mutations (C634R and C634Y) were detected in the patients of two families with MEN2A. Pentagastrin stimulation test was helpful for the diagnosis and follow-up after operative treatment of MEN2A.
Asunto(s)
Neoplasia Endocrina Múltiple Tipo 2a/genética , Mutación Missense , Proteínas Proto-Oncogénicas c-ret/genética , Adulto , Secuencia de Bases , Calcitonina/sangre , Análisis Mutacional de ADN , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Neoplasia Endocrina Múltiple Tipo 2a/diagnóstico , Linaje , Pentagastrina/administración & dosificación , Reacción en Cadena de la Polimerasa , Proto-Oncogenes MasRESUMEN
OBJECTIVE: To detect RET mutations in a rare Chinese big family with Multiple endocrine neoplasia type 2A (MEN2A). METHODS: One MEN2A family, including the proband, have 22 members of two generations, it is a rare big family in modern Chinese families. The DNAs of the 22 members from the family including 4 patients were extracted from blood leukocytes, PCR and gene sequencing of PCR products by an automated DNA sequencer were applied to scan the exon10 and 11 of RET proto-oncogene. Sequencing results were compared with the Pubmed's. Clone sequencing was adopt to further confirm the results, then verifying the novel mutation through the human gene mutation database at the institute of medical genetics in cardiff. Invitrogen biotechnology company (Shanghai) provided the technology of clone sequencing. RESULTS: A novel deletion mutation of D631 (GAC) (del D631) was detected in exon11 of the RET proto-oncogene in 4 MEN2A patients of the family, this rare deletion mutation of D631 (GAC) lead base sequence of TGC(angle)GACGAGCTG change to TGCGAGCTG. Besides 4 MEN2A patients, the son of II6 (the first class relative) was found to be a carrier of delD631 mutation. CONCLUSION: A novel deletion mutation (del D631) of RET proto-oncogene was detected in the family with MEN2A and it has never been reported before in the world. DelD631 may be related to the late onset of MEN2A compared to the cysteine mutations and pheochromocytoma might be the first manifestation prior to the development of MTC.
Asunto(s)
Neoplasia Endocrina Múltiple Tipo 2a/genética , Mutación , Proteínas Proto-Oncogénicas c-ret/genética , Adulto , Secuencia de Bases , China , Análisis Mutacional de ADN , Exones , Femenino , Humanos , Masculino , Linaje , Proto-Oncogenes Mas , Eliminación de SecuenciaRESUMEN
Two new steroidal alkaloids, isoconkuressine and N-formylconessimine, together with 6 known steroidal alkaloids including conkuressine, conessine, isoconessimine, conimine, conarrhimine, and funtudienine, were isolated from the seeds of Holarrhena antidysenteriaca Wall.ex A.DC. Their intrinsic antibacterial activities and synergistic effects with penicillin and vancomycin were analyzed in methicillin sensitive staphylococcus aureus (MSSA) and methicillin resistant staphylococcus aureus (MRSA). Two of the steroidal alkaloids including one new compound (N-formylconessimine) showed potential antibacterial activity and possessed synergistic effects with penicillin and vancomycin, respectively.
Asunto(s)
Alcaloides/farmacología , Antibacterianos/farmacología , Holarrhena/química , Extractos Vegetales/farmacología , Alcaloides/aislamiento & purificación , Antibacterianos/química , Antibacterianos/aislamiento & purificación , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Staphylococcus aureus/efectos de los fármacosRESUMEN
The objective of the study was to enhance the substrate tolerance of Pseudomonas putida nitrilase via atmospheric and room temperature plasma (ARTP) and cell immobilization. The mutant library was constructed by ARTP and rapidly screened by an OPA-TCA microscale reaction. A mutant strain of mut-D3 was obtained and its optimum substrate concentration was improved to 150mM from 100mM. It could accumulate 189g/L nicotinic acid (NA) from 3-cyanopyridine (3-CP), which was increased by 42% compared with that of wild type (WT). Additionally, composite immobilization of mut-D3 was performed and SA-PVA immobilized cells could catalyze 250mM 3-CP each batch with finally accumulating 346g/L NA, while free cells accumulated 175g/L NA. These results indicated that the free or immobilized catalysts of mut-D3 could serve as a good choice for NA production. This is the first report on mutation breeding of nitrilase-producing microorganisms by ARTP.
Asunto(s)
Aminohidrolasas , Pseudomonas putida , Piridinas , TemperaturaRESUMEN
Cation channel of sperm 1 (CATSPER1) is a unique sperm cation channel protein, and essential for sperm function and male fertility. CATSPER1 exclusively expresses in meiotic and postmeiotic spermatogenic cells, thus belongs to the spermatogenesis-specific antigen that escape central tolerance. We have previously demonstrated the immunocontraceptive potential of its transmembrane domains and pore region, and reported the antifertility effects of its B-cell epitopes on male mice. Aiming to develop DNA vaccine targeting CATSPER1 for male contraception, here the whole open reading frame of mouse Catsper1 was cloned into the plasmid pEGFP-N1 to obtain a DNA vaccine pEGFP-N1-Catsper1. The vaccine was confirmed to be transcribed and translated in mouse N2a cell in vitro and mouse muscle tissue in vivo. Intramuscular injection with the vaccine on male mice induced specific immune reaction and caused significant inhibition on sperm hyperactivated motility and progressive motility (P<0.001 for both), and consequently reduced male fertility. The fertility rate of experimental group was 40.9%, which was significant lower (P=0.012) than control group (81.8%). No significant change in mating behavior, sperm production and histology of testis/epididymis was observed. Given that Catsper1 exhibits a high degree of homology among different species, Catsper1 DNA vaccine might be a good strategy for developing an immunocontraceptive vaccine for human and animal use.