RESUMEN

The intention of this study was to analyze the effects of Lactobacillus plantarum (L. plantarum) additive with different nutrient density diets on growth performance, excreta microbiota, nutrient digestibility, gas emission, and meat quality in Ross308-broilers. A total of 576 mixed-sex, 1-d old Ross-308 chicks were randomly allocated to one of four treatment groups with 8 replication and 18 chicks/cage. For a period of 35 days, HD and LD group chicks were fed with commercial corn and soybean meal-based basal diet which contains high and low nutrient density diet, respectively. The other treatment groups LP1 and LP2 chicks were fed with LD+ 0.05% and 0.01 % of L. plantarum, respectively. During day 21 and the overall experimental period, the body weight gain of broilers significantly increased (p<0.05) in HD and L. plantarum groups compared to the LD group. On day 35, broilers fed L. plantarum additive had significantly increased (p<0. 05) the nutrient digestibility of dry matter and nitrogen compared to those fed HD and LD diets. Moreover, dietary inclusion of L. plantarum additive had significantly increased (p<0.05) lactobacillus population and decreased (p>0.05) E. coli and ammonium emission. However, the meat quality traits were not affected by experimental diets. In conclusion, we infer that a low-density diet with 0.1% of L. plantarum additive could serve as an excellent alternative feed additive to enhance the performance of broilers.(AU)

Asunto(s)

Animales , Pollos/metabolismo , Lactobacillus plantarum , Microbiota , Carne , Valor Nutritivo

RESUMEN

The gonadotropin-releasing hormone (GnRH) plays an important role in the control of reproductive functions. Recent studies have reported the occurrence of GnRH molecular variants in numerous species. In this study, the GnRH1 gene from Jinghai yellow chicken was cloned by reverse transcriptase-polymerase chain reaction and transformed into BL21 (DE3) competent cells. The GnRH1 gene and amino acid sequences were subjected to bioinformatic analyses. The GnRH1 gene nucleotide sequence was discovered to be 352 bp long, containing a coding, promoter, and section of the 3'-regions. The GnRH1 gene shared 93, 81, 54, 58, 61, 76, 76, 59, 76, and 66% sequence identity with Meleagris gallopavo, Columba livia, Homo sapiens, Bos taurus, swines, Capra hircus, Ovis aries, Pantholops hodgsonii, Equus caballus, and Rattus norvegicus, respectively. The GnRH1 gene showed conserved domains. The GnRH1 protein was a secreted protein comprising 92 amino acids, with a molecular weight of 10205.6 Da and a theoretical pI of 5.67. Most of the amino acid residues were observed to be hydrophilic, indicating water solubility. The predicted secondary structures of proteins included α-helices (h; 23.08%), ß-extensions (e; 10.92%), and random coils (c; 66.0%). The successful construction of prokaryotic expression vector pET32a-GnRH1 was confirmed by restriction and sequence analysis. SDS-PAGE analysis showed the successful expression of recombinant plasmid in Escherichia coli BL21 (molecular weight = 25-28 kDa). Larger quantities of protein were expressed in supernatant, indicating greater expression in soluble form. Western blot analysis confirmed the expression of the target protein.

Asunto(s)

Proteínas Aviares/genética , Pollos/genética , Escherichia coli/genética , Hormona Liberadora de Gonadotropina/genética , Secuencia de Aminoácidos , Aminoácidos/química , Aminoácidos/genética , Aminoácidos/metabolismo , Animales , Proteínas Aviares/metabolismo , Secuencia de Bases , Western Blotting , Clonación Molecular , Biología Computacional/métodos , ADN Complementario/química , ADN Complementario/genética , Expresión Génica , Hormona Liberadora de Gonadotropina/clasificación , Hormona Liberadora de Gonadotropina/metabolismo , Interacciones Hidrofóbicas e Hidrofílicas , Datos de Secuencia Molecular , Filogenia , Proteínas Recombinantes/metabolismo , Análisis de Secuencia de ADN

RESUMEN

This study aimed to detect micrometastatic tumor cells in the lymph nodes of patients with pN0 lung adenocarcinoma using a combination of thyroid transcription factor-1 (TTF-1) expression and cytokeratin 7 (CK7) expression and to investigate the association of lymph node micrometastasis with the clinicopathological characteristics of patients with lung adenocarcinoma. A total of 54 patients with pN0 lung adenocarcinoma and whose primary tumors were positive for both TTF-1 and CK7 expression were included in this study. In total, 893 lymph nodes were obtained from these 54 patients and were analyzed for micrometastasis by immunohistochemical staining with anti-CK7 and anti-TTF-1 antibodies. CK7- and TTF-1-positive cells were found in the lymph nodes of 9 (16.7%) of 54 patients, and 21 (2.4%) of 893 lymph nodes exhibited positivity for these factors. No cells positive for both CK7 and TTF-1 were detected in the 5 lymph nodes obtained from patients with benign lung tumors. Lymph node micrometastasis was found to be associated with the differentiation grade and primary tumor position (P < 0.05). The detection of lymph node micrometastasis by a combination of CK7 and TTF-1 immunohistochemical staining provides a more accurate assessment of tumor staging for pN0 lung adenocarcinoma.

Asunto(s)

Adenocarcinoma/diagnóstico , Neoplasias Pulmonares/diagnóstico , Ganglios Linfáticos/patología , Adenocarcinoma/genética , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Adenocarcinoma del Pulmón , Adulto , Anciano , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Femenino , Humanos , Inmunohistoquímica , Queratina-7/genética , Queratina-7/metabolismo , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Metástasis Linfática , Masculino , Persona de Mediana Edad , Clasificación del Tumor , Micrometástasis de Neoplasia , Estadificación de Neoplasias , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Factor Nuclear Tiroideo 1 , Factores de Transcripción/genética , Factores de Transcripción/metabolismo

RESUMEN

The identification of human disease-related microRNAs (miRNAs) is important for understanding the pathogenesis of diseases, but to do this experimentally is a costly and time-consuming process. Computational prediction of disease-related miRNA candidates is a valuable complement to experimental studies. It is essential to develop an effective prediction method to provide reliable candidates for subsequent biological experiments. In this study, we constructed a miRNA functional similarity network based on calculation of the functional similarity between each pair of miRNAs. Here, we present a new method (DismiPred) for predicting disease-related miRNA candidates based on the network. This method incorporates functional similarity and common association information to achieve an efficient prediction performance. DismiPred has been successfully shown to recover experimentally validated disease-related miRNAs for 12 common human diseases, with an F-measure ranging from 69.49 to 91.69%. Furthermore, a case study examining breast neoplasms showed that DismiPred could uncover novel disease-related miRNAs. DismiPred is useful for further experimental studies on the involvement of miRNAs in the pathogenesis of diseases.

Asunto(s)

Biología Computacional/métodos , MicroARNs/genética , Algoritmos , Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Femenino , Regulación de la Expresión Génica , Redes Reguladoras de Genes , Humanos , Masculino , MicroARNs/metabolismo , Reproducibilidad de los Resultados , Programas Informáticos

RESUMEN

PURPOSE: This study evaluated the effect of estrogen (E(2)), progesterone (P4), and the combination of them (E(2) + P4) on survival rate, apoptosis, and the expressions of Bcl-2, hsa-let-7a and has-miR-34b in primary ovarian cancer cells to provide new clues for the clinical treatments of ovarian cancer. METHODS: The primary ovarian cancer cells from 60 cases of clinical ovarian cancer tissues were isolated and then cultured. The survival rate of ovarian cancer cells after the treatment of E(2), P4 and E(2) + P4 was analyzed by MTT assay. Cell apoptosis rate and cell cycle were measured by FACS analysis. Moreover, the relative abundance of Bcl-2 and microRNAs (let-7a, miR-34b) expressions were detected by quantitative real-time PCR (qRT-PCR) and Western blotting. RESULTS: Low concentrations of estrogen (10(-10), 10(-8), 10(-6 )mol/L) did not affect the proliferation of ovarian cancer cells. However, the high concentration of estrogen (10(-4 )mol/L) inhibited survival rate of ovarian cancer cells. Progesterone (10(-4 )mol/L) inhibited the proliferation of cancer cells. The combination of estrogen and progesterone significantly inhibited the survival rate of ovarian cancer cells with a time- and dose-dependent manner. High concentration of estrogen combined with progesterone (E(2) + P4) induced apoptosis of ovarian cancer cells. E(2) + P4 promoted the expression of let-7a and miR-34b and reduced the expression of Bcl-2 in ovarian cancer cells. When the expression of let-7a or/and miR-34b was inhibited using miRNA inhibitors, E(2) + P4 treatment did not change the protein level of Bcl-2. CONCLUSION: E(2) + P4 significantly inhibited the cell survival, promoted the cell apoptosis, induced the expression of let-7a and miR-34b, and reduced the expression of Bcl-2 in ovarian cancer cells.

Asunto(s)

Adenocarcinoma de Células Claras , Proliferación Celular/efectos de los fármacos , Estrógenos/farmacología , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Neoplasias Quísticas, Mucinosas y Serosas , Neoplasias Ováricas , Progesterona/farmacología , Teratoma , Adenocarcinoma Mucinoso , Adulto , Anciano , Anciano de 80 o más Años , Apoptosis/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Regulación hacia Abajo , Femenino , Humanos , MicroARNs/efectos de los fármacos , MicroARNs/genética , Persona de Mediana Edad , Proteínas Proto-Oncogénicas c-bcl-2/efectos de los fármacos , Proteínas Proto-Oncogénicas c-bcl-2/genética , Células Tumorales Cultivadas , Regulación hacia Arriba

RESUMEN

Sweet basil, Ocimum basilicum L., is a fragrant herb belonging to the family Lamiaceae. Originated in India 5,000 years ago, sweet basil plays a significant role in diverse cuisines across the world, especially in Asian and Italian cooking. In October 2008, hundreds of plants showing symptoms of leaf spot with nearly 100% incidence were found in polyethylene tunnels at an organic farm in Icheon, Korea. Leaf spots were circular to subcircular, water-soaked, dark brown with grayish center, and reached 10 mm or more in diameter. Diseased leaves defoliated prematurely. The damage purportedly due to this disease has reappeared every year with confirmation of the causal agent made again in 2011. A cercosporoid fungus was consistently associated with disease symptoms. Stromata were brown, consisting of brown cells, and 10 to 40 µm in width. Conidiophores were fasciculate (n = 2 to 10), olivaceous brown, paler upwards, straight to mildly curved, not geniculate in shorter ones or one to two times geniculate in longer ones, 40 to 200 µm long, occasionally reaching up to 350 µm long, 3.5 to 6 µm wide, and two- to six-septate. Conidia were hyaline, acicular to cylindric, straight in shorter ones, flexuous to curved in longer ones, truncate to obconically truncate at the base, three- to 16-septate, and 50 to 300 × 3.5 to 4.5 µm. Morphological characteristics of the fungus were consistent with the previous reports of Cercospora guatemalensis A.S. Mull. & Chupp (1,3). Voucher specimens were housed at Korea University herbarium (KUS). An isolate from KUS-F23757 was deposited in the Korean Agricultural Culture Collection (Accession No. KACC43980). Fungal DNA was extracted with DNeasy Plant Mini DNA Extraction Kits (Qiagen Inc., Valencia, CA). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The resulting sequence of 548 bp was deposited in GenBank (Accession No. JQ995781). This showed >99% similarity with sequences of many Cercospora species, indicating their close phylogenetic relationship. Isolate of KACC43980 was used in the pathogenicity tests. Hyphal suspensions were prepared by grinding 3-week-old colonies grown on PDA with distilled water using a mortar and pestle. Five plants were inoculated with hyphal suspensions and five plants were sprayed with sterile distilled water. The plants were covered with plastic bags to maintain a relative humidity of 100% for 24 h and then transferred to a 25 ± 2°C greenhouse with a 12-h photoperiod. Typical symptoms of necrotic spots appeared on the inoculated leaves 6 days after inoculation, and were identical to the ones observed in the field. C. guatemalensis was reisolated from symptomatic leaf tissues, confirming Koch's postulates. No symptoms were observed on control plants. Previously, the disease was reported in Malawi, India, China, and Japan (2,3), but not in Korea. To our knowledge, this is the first report of C. guatemalensis on sweet basil in Korea. Since farming of sweet basil has recently started on a commercial scale in Korea, the disease poses a serious threat to safe production of this herb, especially in organic farming. References: (1) C. Chupp. A Monograph of the Fungus Genus Cercospora. Ithaca, NY, 1953. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology & Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , May 5, 2012. (3) J. Nishikawa et al. J. Gen. Plant Pathol. 68:46, 2002.

RESUMEN

To solve the class imbalance problem in the classification of pre-miRNAs with the ab initio method, we developed a novel sample selection method according to the characteristics of pre-miRNAs. Real/pseudo pre-miRNAs are clustered based on their stem similarity and their distribution in high dimensional sample space, respectively. The training samples are selected according to the sample density of each cluster. Experimental results are validated by the cross-validation and other testing datasets composed of human real/pseudo pre-miRNAs. When compared with the previous method, microPred, our classifier miRNAPred is nearly 12% more accurate. The selected training samples also could be used to train other SVM classifiers, such as triplet-SVM, MiPred, miPred, and microPred, to improve their classification performance. The sample selection algorithm is useful for constructing a more efficient classifier for the classification of real pre-miRNAs and pseudo hairpin sequences.

Asunto(s)

Biología Computacional/métodos , MicroARNs/análisis , Algoritmos , Secuencia de Bases , Humanos , MicroARNs/química , Conformación de Ácido Nucleico

RESUMEN

OBJECTIVE: To evaluate tubal patency using hysterosalpingography after clinical treatment of tubal pregnancy. METHOD: Of 80 patients who underwent hysterosalpingography after clinical treatment of tubal pregnancy from April 1994 to February 2002, 30 were treated with a single 50 mg/m(2) dose of methotrexate intramuscularly (n=30) and 50 were followed up expectantly. RESULTS: Patency of the ipsilateral tube was 84% after methotrexate treatment and 78% after expectant management. In addition, contralateral tubal patency was 97% after methotrexate treatment and 92% after expectant management. There were no statistically significant differences between the groups. CONCLUSION: Findings from this study suggest similar tubal patency rates after methotrexate treatment and expectant management.

Asunto(s)

Trompas Uterinas/fisiología , Embarazo Tubario/cirugía , Abortivos no Esteroideos/uso terapéutico , Femenino , Humanos , Histerosalpingografía , Metotrexato/uso terapéutico , Embarazo , Embarazo Tubario/tratamiento farmacológico , Estudios Prospectivos , Recuperación de la Función

RESUMEN

BACKGROUND: Two natural rubber latex proteins, Hev b 1 and Hev b 3, have been described in spina bifida (SB)-associated latex allergy. OBJECTIVE: The aim of this study was to clone and express Hev b 3 and to obtain the immunologic active and soluble recombinant allergen for diagnosis of SB-associated latex allergy. METHODS: A complementary DNA (cDNA) coding for Hev b 3 was amplified from RNA of fresh latex collected from Malaysian rubber trees (Hevea brasiliensis). PCR primers were designed according to sequences of internal peptide fragments of natural (n) Hev b 3. The 5'-end sequence was obtained by specific amplification of cDNA ends. The recombinant (r) Hev b 3 was produced in Escherichia coli as a 6xHis tagged protein. Immunoblotting and inhibition assays were performed to characterize the recombinant allergen. RESULTS: An Hev b 3 cDNA clone of 922 bp encoding a protein of 204 amino acid residues corresponding to a molecular weight of 22.3 kd was obtained. In immunoblots 29/35, latex-allergic patients with SB revealed IgE binding to rHev b 3, as did 4 of 15 of the latex-sensitized group. The presence of all IgE epitopes on rHev b 3 was shown by its ability to abolish all IgE binding to nHev b 3. Hev b 3 is related to Hev b 1 by a sequence identity of 47%. Cross-reactivity between these 2 latex allergens was illustrated by the large extent of inhibition of IgE binding to nHev b 1 by rHev b 3. CONCLUSION: rHev b 3 constitutes a suitable in vitro reagent for the diagnosis of latex allergy in patients with SB. The determination of the full sequence of Hev b 3 and the production of the recombinant allergen will allow the epitope mapping and improve diagnostic reagents for latex allergy.

Asunto(s)

Alérgenos/inmunología , Hipersensibilidad al Látex/inmunología , Látex/inmunología , Proteínas de Plantas/inmunología , Proteínas Recombinantes de Fusión/inmunología , Disrafia Espinal/inmunología , Adolescente , Adulto , Alérgenos/genética , Alérgenos/aislamiento & purificación , Secuencia de Aminoácidos , Antígenos de Plantas , Secuencia de Bases , Niño , Clonación Molecular , ADN de Plantas , Euphorbiaceae/genética , Estudios de Evaluación como Asunto , Femenino , Expresión Génica , Humanos , Immunoblotting , Inmunoglobulina E/inmunología , Hipersensibilidad al Látex/sangre , Hipersensibilidad al Látex/complicaciones , Masculino , Datos de Secuencia Molecular , Proteínas de Plantas/genética , Proteínas de Plantas/aislamiento & purificación , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/aislamiento & purificación , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Disrafia Espinal/sangre , Disrafia Espinal/complicaciones

RESUMEN

Biochemical evidence reported so far suggests that rubber synthesis takes place on the surface of rubber particles suspended in the latex of Hevea brasiliensis. We have isolated and characterized a cDNA clone that encodes a protein tightly bound on a small rubber particle. We named this protein small rubber particle protein (SRPP). Prior to this study, this protein was known as a latex allergen, and only its partial amino acid sequence was reported. Sequence analysis revealed that this protein is highly homologous to the rubber elongation factor and the Phaseolus vulgaris stress-related protein. Southern and Northern analyses indicate that the protein is encoded by a single gene and highly expressed in latex. An allergenicity test using the recombinant protein confirmed that the cloned cDNA encodes the known 24-kDa latex allergen. Neither ethylene stimulation nor wounding changed the transcript level of the SRPP gene in H. brasiliensis. An in vitro rubber assay showed that the protein plays a positive role in rubber biosynthesis. Therefore, it is likely that SRPP is a part of the rubber biosynthesis machinery, if not the rubber polymerase, along with the rubber elongation factor.

Asunto(s)

Alérgenos , Euphorbiaceae/genética , Genes de Plantas , Proteínas de Plantas/genética , Goma/metabolismo , Árboles/genética , Secuencia de Aminoácidos , Antígenos de Plantas , Secuencia de Bases , Dosificación de Gen , Expresión Génica , Hipersensibilidad al Látex , Datos de Secuencia Molecular , Proteínas de Plantas/inmunología , Proteínas de Plantas/metabolismo , Homología de Secuencia de Aminoácido , Terpenos/metabolismo

Asunto(s)

Inmunohistoquímica , Enfermedades Periodontales , Cicatrización de Heridas

Asunto(s)

Raíz del Diente , Factor de Crecimiento Derivado de Plaquetas , Regeneración Tisular Dirigida , Defectos de Furcación