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1.
Fish Shellfish Immunol ; 93: 144-152, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31326584

RESUMO

Phosphoglycerate kinase (EC 2.7.2.3, PGK) catalyses the reversible transfer of a phosphate group from 1,3-diphosphoglyceric acid and ADP to produce 3-phosphoglyceric acid and ATP, which represents the initial production of ATP during glycolysis; therefore, PGK is a key enzyme in the energy metabolism. To study the role of PGK in the resistance to WSSV infection in shrimp, the full-length cDNA of the PGK gene (LvPGK) from Litopenaeus vannamei was obtained by using homology cloning and RACE amplification. The tissue distribution of LvPGK and its expression changes in the main immune tissues after WSSV stimulation were obtained by quantitative real-time PCR. Furthermore, RNA interference (RNAi) was used to study the role of LvPGK in shrimp defending against WSSV infection. The results showed that the full-length cDNA sequence of LvPGK was 1855 bp, contained a 1248 bp open reading frame (ORF) encoding 415 amino acids, and included a conserved PGK domain. LvPGK presented ubiquitous expression in most examined tissues, with the most predominant expression in the muscle and the weakest expression in the intestine. LvPGK transcripts could be induced in the hemocytes and hepatopancreas by injection with WSSV. Both the replication of WSSV and the shrimp cumulative mortality decreased significantly after LvPGK knockdown (P < 0.01). After challenging LvPGK RNAi shrimp with WSSV, the concentration of glucose in the hepatopancreas and muscle tissue did not show significant change; however, the content of pyruvate and lactate decreased significantly (P < 0.05). Moreover, significant decreases in the expression levels of crustin, ALF1, ALF2 and ALF3 were also detected. The results suggested that LvPGK might be involved in WSSV replication by increasing host aerobic and anaerobic metabolism.


Assuntos
Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Penaeidae/genética , Penaeidae/imunologia , Fosfoglicerato Quinase/genética , Fosfoglicerato Quinase/imunologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Sequência de Bases , Perfilação da Expressão Gênica , Penaeidae/enzimologia , Fosfoglicerato Quinase/química , Filogenia , Alinhamento de Sequência , Vírus da Síndrome da Mancha Branca 1/fisiologia
2.
Fish Shellfish Immunol ; 91: 275-283, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31125663

RESUMO

Glycogen phosphorylase (GP, EC 2.4.1.1) catalyze the rate-limiting step in glycogenolysis in animals, forming glucose-1-phosphate from the terminal alpha-1,4-glycosidic bond. Therefore, GP plays a crucial role in carbohydrate metabolism. In the present study, the full-length cDNA sequence of GP (LvGP) was cloned from shrimp, Litopenaeus vannamei. The obtained 3242-bp LvGP cDNA sequence included a 5'-terminal untranslated region (UTR) of 48 bp, an open reading frame (ORF) of 2559 bp encoding a polypeptide of 852 amino acids (aa) and a 3'-UTR of 635 bp. The predicted LvGP protein sequence contained a typical phosphorylase domain (113-829 aa) and shared 72%-97% identities with GP from other species. Phylogenetic analysis revealed that LvGP showed the closest relationship with GP from Marsupenaeus japonicus. Tissue expression profiles showed that LvGP existed in most examined tissues, with the most predominant expression in the brain, followed by the muscles and stomach. LvGP transcripts in hepatopancreas and hemocytes were up regulated after the WSSV challenge. Furthermore, the role of LvGP in shrimp defending against WSSV infection was investigated by RNA interference (RNAi). Our findings showed that WSSV proliferation and shrimp accumulative mortality increased significantly after LvGP RNAi (P < 0.01). The glycogen, glucose, and pyruvate content decreased in GP RNAi shrimp after WSSV injection, however, the lactate and ATP concentration enhanced. Moreover, lectin and anti-lipopolysaccharide factor2 (ALF2) were induced in LvGP silencing shrimp after WSSV infection, whereas the expression levels of crustin, ALF1 and ALF3 decreased. Our results suggested that the LvGP might play a crucial role in shrimp defending against WSSV infection by regulating metabolism and affecting the anti-infectious gene expression.


Assuntos
Regulação da Expressão Gênica/imunologia , Glicogênio Fosforilase/genética , Glicogênio Fosforilase/imunologia , Imunidade Inata/genética , Penaeidae/genética , Penaeidae/imunologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Sequência de Bases , Perfilação da Expressão Gênica , Glicogênio Fosforilase/química , Filogenia , Interferência de RNA , Regulação para Cima , Vírus da Síndrome da Mancha Branca 1/fisiologia
3.
Fitoterapia ; 128: 20-25, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29723562

RESUMO

Five new amide alkaloids (1-3, 5-6) were isolated from the seeds of Clausena lansium together with one new natural product (4) and four known analogues (7-10). The structures of the new amide alkaloids were elucidated based on a comprehensive spectroscopic data analysis including 1D and 2D NMR as well as HRESIMS, and by comparison with the literature. The bioactivity results showed that compound 8 expressed potent nematicidal activity against Panagrellus redivevus, with IC50 value of 0.12 mM, while compounds 3 and 5 presented moderate nematicidal activity with IC50 values of 2.75 and 3.93 mM, respectively (abamectin as the positive control with IC50 value of 1.05 mM).


Assuntos
Alcaloides/isolamento & purificação , Amidas/isolamento & purificação , Anti-Helmínticos/isolamento & purificação , Clausena/química , Sementes/química , Alcaloides/farmacologia , Amidas/farmacologia , Animais , Anti-Helmínticos/farmacologia , Estrutura Molecular , Tylenchida/efeitos dos fármacos
4.
J Asian Nat Prod Res ; 20(1): 55-61, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28597686

RESUMO

Three new flavanoids, (2R)-7,4'-dihydroxy-8-methylflavan (1), (2R)-7,4'-dihydroxy-6-methylflavan (2), and (3R)-7,3',4'-trihydroxyhomoisoflavan (3), together with seven known compounds (4-10), were isolated from artificially induced dragon's blood of Dracaena cambodiana, and their structures were determined based on HR-ESI-MS and extensive spectroscopic techniques (UV, IR, 1D-, and 2D-NMR). Compound 2 exhibited weak cytotoxicity against BEL-7402 cells line with the IC50 value of 39.2 µM. In addition, compound 3 showed significant acetylcholinesterase (AChE) inhibitory activity.


Assuntos
Inibidores da Colinesterase/isolamento & purificação , Inibidores da Colinesterase/farmacologia , Dracaena/química , Flavonoides/isolamento & purificação , Inibidores da Colinesterase/química , Flavonoides/química , Flavonoides/farmacologia , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Extratos Vegetais/química
5.
Anim Sci J ; 89(1): 52-59, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28960617

RESUMO

The Kunming dog is the first and only working dog breed from China to be recognized worldwide. As a domestic working dog, its excellent working performance has been well established; however, its normal reproductive parameters are not well understood. Therefore, this study was conducted to document the main reproductive parameters of this purebred working dog in field breeding conditions. Data on 1004 heats (753 with mating) from 203 bitches between 2008 to 2014, were collected and analyzed. The pregnancy rate and whelping rate was 79.42% and 75.30%, respectively. Finally, for 567 litters (4298 puppies), the mean litter size was 7.19 ± 0.12 puppies (range 1-15). The mean gestation period and birth weight were approximately 61.64 ± 0.10 days and 407.25 ± 1.21 g. The mean sex ratio was 1.03 males to 1.00 female. Estrus occurred throughout the year with no significant differences between seasons and months (P > 0.05), which confirms that Kunming dogs are non-seasonal breeders; births occurred in every month of the year. Pregnant bitches exhibited significantly longer inter-estrus intervals than non-pregnant bitches (220.85 ± 2.05 vs. 180.19 ± 2.94 days, P < 0.05). Bitch parity influenced litter size, and the gestation length and birth weight of the puppies were negatively affected by litter size. This study helps elucidate the reproductive potential of this breed and provides reference values for reproductive performance in the Kunming dog.


Assuntos
Cães/fisiologia , Reprodução/fisiologia , Animais , Peso ao Nascer , Diterpenos , Estro/fisiologia , Feminino , Indóis , Tamanho da Ninhada de Vivíparos , Masculino , Gravidez , Estudos Retrospectivos
6.
Fitoterapia ; 121: 1-5, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28648631

RESUMO

Dragon's blood is a rare traditional medicine used in many countries. Because of its various therapeutic uses, its requirement is increasing dramatically, while formation of dragon's blood need at least several years, so many researchers tried to induce the formation of dragon's blood artificially, but the chemical constituents of artificially induced dragon's blood were rarely studied. In our research, twenty constituents, including three new ones, were isolated from artificially induced dragon's blood of Dracaena cambodiana, five of which are the same with those from natural dragon's blood. In addition, six compounds exhibited cytotoxic activities, and eleven compounds demonstrated antibacterial activities. Artificially induced dragon's blood is expected to be the substituent of natural dragon's blood.


Assuntos
Dracaena/química , Flavonoides/química , Extratos Vegetais/química , Resinas Vegetais/química , Antibacterianos/química , Antibacterianos/isolamento & purificação , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/isolamento & purificação , Linhagem Celular Tumoral , Flavonoides/isolamento & purificação , Humanos , Estrutura Molecular , Extratos Vegetais/isolamento & purificação
7.
Anim Sci J ; 88(8): 1232-1238, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28078817

RESUMO

The aim of this study was to evaluate fertility and sex ratios after artificial insemination in dogs under field conditions. Semen was cryopreserved as unsorted (control) or was separated into X- and Y-chromosome-bearing sperm using a cell sorter. Sixty female dogs were inseminated with frozen-thawed spermatozoa of 100 × 106 unsorted (a dose in practice) and 4 × 106 sorted (X and Y group, respectively). A total of 20 dogs became pregnant and 126 puppies were born from the three groups. The percentage of parturition was similar for the X (5/20; 25.0%) and Y (4/20; 20.0%) group (P > 0.05), but lower than controls (11/20; 55.0%) (P < 0.05). Ultimately 28 out of the 32 puppies produced from X group were female (87.5%) and 19/22 (86.4%) puppies of Y group were male. In contrast, sex ratio (51.4% to 48.6%) in the control was significantly different from the X, Y group (P < 0.05). However, male and female puppies in the control had similar birth weights and weaning weights to those from the X and Y groups. This preliminary information indicated that normal puppies of predicted sex can be produced with low numbers of sorted cryopreserved dog spermatozoa at a farm level, making sperm-sexing technology potentially applicable for elite breeding units.


Assuntos
Criopreservação , Fertilidade , Congelamento , Inseminação Artificial , Parto , Prenhez , Preservação do Sêmen , Pré-Seleção do Sexo/métodos , Razão de Masculinidade , Espermatozoides , Animais , Cães , Feminino , Masculino , Gravidez
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