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This article reviews the relevant studies on the efficacy and safety of influenza, pneumococcal and COVID-19 vaccination among tumor patients worldwide in recent years. By combing and analyzing the retrieved literature, the results show that influenza and pneumococcal vaccination can significantly reduce the morbidity and hospitalization rate of infectious diseases in tumor patients, reduce the risk of cardiovascular events and death, and significantly improve survival prognosis. COVID-19 vaccination can also protect tumor patients, especially those who have completed full dose vaccination. Authoritative guidelines and consensuses worldwide all recommend that tumor patients receive influenza, pneumococcal and COVID-19 vaccines. We should carry out relevant researches, as well as take effective measures to strengthen patient education, so that tumor patients can fully experience the health protection brought by the vaccine to this specific group.
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COVID-19 , Vacinas contra Influenza , Influenza Humana , Neoplasias , Humanos , Influenza Humana/prevenção & controle , Vacinas contra COVID-19 , COVID-19/prevenção & controle , Vacinas contra Influenza/uso terapêutico , Vacinação , Vacinas Pneumocócicas/uso terapêutico , Streptococcus pneumoniaeRESUMO
Objective: To analyze the relationship between renin-angiotensin-aldosterone system (RAAS) gene polymorphisms and susceptibility to essential hypertension (EH) in military secret service personnel. Methods: In October 2019, military secret service personnel (162 people) who were recuperating in a sanatorium from January to December 2017 were selected as the research subjects, all of whom were Han and male. The patients (79 people) who were diagnosed with EH according to the diagnostic criteria of hypertension in the "Chinese Guidelines for the Prevention and Treatment of Hypertension" (2016 Revised Edition) were the case group, and the people with normal blood pressure (83 people) were the control group. Occupational epidemiological investigation was conducted, 5 ml of fasting cubital venous blood was collected, genomic DNA was extracted by phenol-chloroform method, and RAAS gene polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism method. The distribution differences of genotype and allele frequency between groups were compared, and the relationship between genotype, allele frequency and EH was analyzed. Results: The average age of military secret service personnel was (38.2±5.3) years old, and there was no statistical significance in the average age and the age distribution over 40 years old of the case group and the control group (P>0.05) . There were significant differences in the distribution of AGT gene M235T locus, ACE gene I/D polymorphism genotype and allele between the case group and the control group (P<0.05) . The TT genotype with AGT gene M235T locus (OR=3.28, 95%CI: 1.21-8.91) and DD genotype with ACE gene (OR=2.86, 95%CI: 1.17-7.00) were risk factors for EH in military secret service personnel. Conclusion: The TT genotype of AGT gene M235T and the DD genotype of ACE gene may be the susceptibility genotypes of military secret service personnel for EH.
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Hipertensão , Militares , Adulto , Hipertensão Essencial , Frequência do Gene , Genótipo , Humanos , Hipertensão/genética , Masculino , Peptidil Dipeptidase A/genética , Polimorfismo Genético , Sistema Renina-Angiotensina/genéticaRESUMO
Intestinal epithelial cells (IEC) are important parts of the mucosal barrier, whose function can be impaired upon various injury factors such as lipopolysaccharide. Although food-derived exosomes are preventable against intestinal barrier injuries, there have been few studies on the effect of yak milk-derived exosomes and the underlying mechanism that remains poorly understood. This study aimed to characterize the effect of exosomal proteins derived from yak and cow milk on the barrier function of IEC-6 treated with lipopolysaccharide and the relevant mechanism involved. Proteomics study revealed 392 differentially expressed proteins, with 58 higher expressed and 334 lower expressed in yak milk-derived exosomes than those in cow exosomes. Additionally, the top 20 proteins with a relatively consistent higher expression in yak milk exosomes than cow milk exosomes were identified. Protein CD46 was found to be a regulator for alleviating inflammatory injury of IEC-6. In vitro assay of the role of yak milk exosomes on survival of IEC-6 in inflammation by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay confirmed the effectiveness of yak milk exosomes to increase IEC-6 survival up to 18% for 12 h compared with cow milk exosomes (up to 12%), indicating a therapeutic effect of yak milk exosomes in the prevention of intestinal inflammation. Furthermore, yak and cow milk exosomes were shown to activate the PI3K/AKT/C3 signaling pathway, thus promoting IEC-6 survival. Our findings demonstrated an important relationship between yak and cow milk exosomes and intestinal inflammation, facilitating further understanding of the mechanisms of inflammation-driven epithelial homeostasis. Interestingly, compared with cow milk exosomes, yak milk exosomes activated the PI3K/AKT/C3 signaling pathway more to lower the incidence and severity of intestine inflammation, which might represent a potential innovative therapeutic option for intestinal inflammation.
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Doenças dos Bovinos , Exossomos , Animais , Bovinos , Feminino , Inflamação/veterinária , Intestinos , Lipopolissacarídeos , Leite , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-aktRESUMO
Intestinal epithelial cells (IEC) act as an important intestinal barrier whose function can be impaired upon induction by hypoxia. Although intestinal barrier injuries are preventable by milk-derived exosomal microRNAs (miRNAs), the underlying mechanism remains poorly understood. This study aimed to characterize the effect of yak and cow milk-derived exosomal miRNA on the barrier function of IEC-6 under hypoxic conditions, and explore the mechanism of yak milk exosomal miRNA to relieve the hypoxia stress. First, by Illumina HiSeq 2500 (Illumina Inc., San Diego, CA) sequencing, the miRNA expression was systematically screened, and differential expression of 130 miRNAs was identified with 51 being upregulated and 79 downregulated in yak and cow milk-derived exosomes. Furthermore, the top 20 miRNAs that had a relatively consistent high expression in yak milk exosome were identified, and bta-miR-34a was found to be an effective regulator for alleviating hypoxic injury of IEC-6. In vitro assay of the role of bta-miR-34a on survival of IEC-6 in hypoxia by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) confirmed its effectiveness to significantly increase IEC-6 survival up to 13% for 12 h, and up to 9.5% for 24 h. Investigation on the regulatory relationship between bta-miRNA-34a and the hypoxia-inducible factor/apoptosis signaling pathway provided insights into the possible mechanisms by which bta-miR-34a activated the hypoxia-inducible factor and apoptosis signaling pathway, thus promoting IEC-6 survival. The results of this study suggest an important relationship between miRNA expression and intestine barrier integrity, which facilitated further understanding of the physiological function of yak and cow milk exosomal miRNAs, as well as mechanisms of hypoxia-driven epithelial homeostasis.
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Bovinos , Hipóxia Celular/fisiologia , Células Epiteliais/fisiologia , Intestinos/citologia , MicroRNAs/fisiologia , Leite/química , Doença da Altitude/fisiopatologia , Doença da Altitude/veterinária , Animais , Apoptose , Doenças dos Bovinos/fisiopatologia , Contagem de Células/veterinária , Linhagem Celular , Proliferação de Células/fisiologia , Regulação para Baixo , Exossomos/química , Feminino , HumanosRESUMO
Insulin-like peptides (ILPs) including insulin, insulin-like growth factor (IGF) and relaxin are evolutionarily conserved hormones in metazoans, and they are involved in diverse physiological processes. The migratory brown planthopper (BPH), Nilaparvata lugens, encodes four ILP genes (Nlilp1, Nlilp2, Nlilp3 and Nlilp4) but their physiological roles are largely unknown. Sequence analysis showed that NlILP1 contained a relaxin-specific G protein-coupled receptor-binding motif and a variant motif of cysteine residues, and NlILP2 and NlILP4 resembled vertebrate IGFs. RNA interference (RNAi)-mediated gene silencing showed that depletion of each of Nlilp1, 2 and 3 significantly delayed the developmental duration of nymphs, and this effect could be exacerbated by double or triple gene depletion. Depletion of Nlilp1, Nlilp2 or Nlilp3 induces the accumulation of glucose, trehalose and glycogen, which is contradictory to depletion of the insulin receptor (NlInR1) in the BPH. Depletion of Nlilp1 significantly enhanced starvation resistance in both females and males although its extent was smaller than NlInR1 depletion. A parental RNAi assay showed that depletion of each of Nlilp1-4 dramatically impaired female fecundity. These findings indicate that NlILP1-4 have redundant and distinct roles in physiological processes in the BPH, thereby enhancing our understanding of the contribution of each NlILP to the ecological success of this species in natural habitats.
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Hemípteros/genética , Proteínas de Insetos/genética , Sequência de Aminoácidos , Animais , Feminino , Hemípteros/crescimento & desenvolvimento , Hemípteros/metabolismo , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Masculino , Ninfa/genética , Ninfa/crescimento & desenvolvimento , Ninfa/metabolismo , Interferência de RNARESUMO
Objective: To investigate the protective effect of somatostatin (SS) on acute kidney injury (AKI) of paraquat (PQ) poisoned mice and its mechanism. Methods: From December 2017 to April 2018, a total of 48 SPF male BALB/C mice were selected and randomly divided into 4 groups, with 12 mice in each group: Control group, SS group (20 mg/kg SS was injected 1 hour before and 3 hours after gavage with normal saline) , PQ group (2% PQ 60 mg/kg by gavage) and PQ+SS group (Intragastric administration was performed with 2% PQ solution of 60 mg/kg, and 20 mg/kg SS was administered 1 h before and 3 h after intragastric administration) , 12 mice in each group were observed for the general situation and behavioral effects. After 24 hours of modeling, mice were sacrificed.Then blood was extracted after eyeball was removed, and both kidneys were removed by laparotomy. Serum IL-6, TNF-α and MPO levels were determined by ELISA. The characteristic pathological changes of toxic renal tubular injury were observed under light microscope and scored accordingly. The changes of NF-κB expression were detected by Western-Blot, SOD, Caspase-3 and malondialdehyde (MDA) were detected by chemical colorimetry. Results: Mice in Control group and SS group showed normal general conditions and behaviors; Mice in PQ group were significantly worse than those in Control group, showing decreased feeding and activity, dry fur, hair shedding and listless spirit; The above symptoms in the mice of PQ+SS group were alleviated compared with the PQ group. Under the light microscope, the renal tissue structure of PQ group was obviously disordered and severely damaged, and the nephropathy score was (6.14±0.72) . The performance of PQ+SS group under light microscope was improved compared with PQ group, and nephropathy score (4.36±0.42) decreased (P<0.05) . Compared with the Control group, serum TNF-α (39.89±3.32) pg/ml, IL-6 (77.29±4.77) pg/ml, renal NF-κB (2.29±0.097) , MPO (0.31±0.017) µg/ml, MDA (0.91±0.03) mmol/mg prot, and Caspase-3 (376.51±8.24) % levels were significantly increased in the PQ group, while the level of renal SOD (2.36±0.73) U/mg prot was significantly decreased (P<0.05) . Compared with the PQ group, serum TNF-α (33.82±1.57) pg/ml, IL-6 (58.49±5.89) pg/ml, renal NF-κB (0.84±0.05) , MPO (0.22±0.01) µg/ml, MDA (0.72±0.05) mmol/mg prot, Caspase-3 (327.32±21.93) % decreased significantly, and renal SOD (4.90±0.81) U/mg prot increased significantly in the PQ+SS group (P<0.05) . Conclusion: PQ poisoning can lead to AKI in mice, while SS can reduce AKI caused by PQ poisoning, improve the general survival state of PQ poisoned mice, and play a certain protective role in kidney injury caused by PQ poisoning, which may be achieved by inhibiting oxidative stress response, inflammatory response and apoptosis caused by poisoning.
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Paraquat/toxicidade , Somatostatina/metabolismo , Animais , Rim , Pulmão , Masculino , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Objective: To investigate the effects of mono-carbonyl analogues of curcumin (L6H21) on paraquat (PQ) -induced injury in HK-2 cell line and explore its underlying mechanisms. Methods: Cultured HK-2 cells were challenged by PQ with or without L6H21 treatment. Cell viability and apoptosis were determined by CCK-8 assay and flow cytometry, respectively. Gene expressions and protein levels of apoptotic and inflammatory factors were assessed by RT-PCR, ELISA, and western blot. Intracellular ROS production was detected by DCFH-DA staining. Superoxide dismutase (SOD) and malondialdehyde (MDA) were examined by chemical colorimetry. Results: 1) PQ challenge significantly inhibited HK-2 cells proliferation, which was prevented by L6H21 administration. PQ dramatically induced HK-2 apoptosis evidenced by increasing expressions of caspase-9, caspase-3 and Bax, while decreasing Bcl-2 level. However, PQ induced these apoptotic effects in HK-2 cells were reversed by L6H21. Similarly, PQ exposure obviously enhanced activity of NF-κB and levels of cytokines (TNF-αãIL-6) in HK-2 cells, which was inhibited by L6H21. Furthermore, administration of L6H21 inhibited PQ induced ROS and MDA production, and promoted SOD level in HK-2 cells. Conclusion: L6H21 administration inhibits PQ-induced apoptosis in HK-2 cells possibly by reducing inflammation and oxidative damage.
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Apoptose/efeitos dos fármacos , Curcumina/farmacologia , Inflamação/prevenção & controle , Estresse Oxidativo/efeitos dos fármacos , Paraquat/toxicidade , Linhagem Celular , Humanos , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismoRESUMO
This paper reviews the domestic and foreign studies published in 2020 on the application of influenza vaccine in populations at high risk. The importance of influenza vaccination in population at high risk has been proved by larger sample, multicentre, high-quality evidence-based studies. Influenza vaccination is the most cost-effective measure to prevent influenza. However, the coverage rate of influenza vaccine is very low in China, it is necessary to strengthen the health education to promote influenza vaccination in different populations. It is recommended to give influenza vaccination to the population in whom influenza vaccination has been proven safe and effective before influenza season. Research of the safety, efficiency and cost-effectiveness of influenza vaccine should be accelerated for the populations in whom such data are lacking or insufficient.
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Vacinas contra Influenza , Influenza Humana , China , Custos e Análise de Custo , Humanos , Vacinas contra Influenza/efeitos adversos , Influenza Humana/epidemiologia , Influenza Humana/prevenção & controle , VacinaçãoRESUMO
A comprehensive review of the research of the effectiveness of influenza vaccine and 23 valent pneumococcal polysaccharide vaccine (PPV23) in patients with chronic obstructive pulmonary disease (COPD) both at home and abroad in recent years showed that influenza vaccine and PPV23 immunization can significantly reduce the risk for influenza and pneumonia in COPD patients, and reduce the acute exacerbation of disease and related hospitalization. In particular, the influenza vaccination can also reduce the risk for ischemic heart disease, acute coronary syndrome, ventricular arrhythmia, lung cancer, dementia and death in the patients, and the immunization of both vaccines has a more significant protective effect. It is recommended by authoritative guidelines both at home and abroad that COPD patients can receive influenza vaccine and PPV23. At present, the coverage of domestic influenza and pneumococcal vaccines are low, and there are less studies in the applications of both vaccines in patients with COPD. Effective measures should be taken to strengthen the health education and increase the vaccination coverage. Additionally, the clinical research of influenza vaccine and PPV23 for COPD patients, especially the analysis on clinical benefit of immunization of both vaccines, should be further strengthened to effectively improve the survival and prognosis of COPD patients.
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Vacinas contra Influenza , Influenza Humana , Doença Pulmonar Obstrutiva Crônica , Humanos , Vacinas contra Influenza/uso terapêutico , Influenza Humana/prevenção & controle , Vacinas Pneumocócicas , VacinaçãoRESUMO
Objective: To evaluate the safety of two inactivated COVID-19 vaccines in a large-scale emergency use. Methods: Based on the "Vaccination Information Collection System", the incidence data of adverse reactions in the population vaccinated with the inactivated COVID-19 vaccines developed by Beijing Institute of Biological Products Co., Ltd and Wuhan Institute of Biological Products Co., Ltd, respectively, in emergency use were collected, and the relevant information were analyzed with descriptive epidemiological and statistical methods. Results: By December 1, 2020, the vaccination information of 519 543 individuals had been collected. The overall incidence rate of adverse reactions was 1.06%, the incidence rate of systemic adverse reactions was 0.69% and the incidence rate of local adverse reactions was 0.37%. The main systemic adverse reactions included fatigue, headache, fever, cough and loss of appetite with the incidence rates of 0.21%, 0.14%, 0.06%, 0.05% and 0.05%, respectively; the main local adverse reactions were injection site pain and injection site swelling with the incidence rates of 0.24% and 0.05%, respectively. Conclusion: The two inactivated COVID-19 vaccines by Beijing Institute of Biological Products Co., Ltd and Wuhan Institute of Biological Products Co., Ltd showed that in the large-scale emergency use, the incidence rate of general reactions was low and no serious adverse reactions were observed after the vaccinations, demonstrating that the vaccines have good safety.
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COVID-19 , Vacinas , Vacinas contra COVID-19 , Humanos , SARS-CoV-2 , Vacinação , Vacinas/efeitos adversos , Vacinas de Produtos InativadosRESUMO
The crystal structure of the human class III chi chi alcohol dehydrogenase (ADH) in a binary complex with NAD+(gamma) was solved to 2.7 A resolution by molecular replacement with human class I beta1 beta1 ADH. chi chi ADH catalyzes the oxidation of long-chain alcohols such as omega-hydroxy fatty acids as well as S-hydroxymethyl-glutathione, a spontaneous adduct between formaldehyde and glutathione. There are two subunits per asymmetric unit in the chi chi ADH structure. Both subunits display a semi-open conformation of the catalytic domain. This conformation is half-way between the open and closed conformations described for the horse EE ADH enzyme. The semi-open conformation and key changes in elements of secondary structure provide a structural basis for the ability of chi chi ADH to bind S-hydroxymethyl-glutathione and 10-hydroxydecanoate. Direct coordination of the catalytic zinc ion by Glu68 creates a novel environment for the catalytic zinc ion in chi chi ADH. This new configuration of the catalytic zinc is similar to an intermediate for horse EE ADH proposed through theoretical computations and is consistent with the spectroscopic data of the Co(II)-substituted chi chi enzyme. The position for residue His47 in the chi chi ADH structure suggests His47 may function both as a catalytic base for proton transfer and in the binding of the adenosine phosphate of NAD(H). Modeling of substrate binding to this enzyme structure is consistent with prior mutagenesis data which showed that both Asp57 and Arg115 contribute to glutathione binding and that Arg115 contributes to the binding of omega-hydroxy fatty acids and identifies additional residues which may contribute to substrate binding.
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Aldeído Oxirredutases/química , Aldeído Oxirredutases/metabolismo , Álcool Desidrogenase/química , Álcool Desidrogenase/classificação , Álcool Desidrogenase/metabolismo , Sítios de Ligação , Cristalografia por Raios X , Humanos , Modelos Moleculares , Dados de Sequência Molecular , NAD/química , NAD/metabolismo , Conformação Proteica , Zinco/metabolismoRESUMO
Integration of retroviral cDNA is a necessary step in viral replication. The virally encoded integrase protein and DNA sequences at the ends of the linear viral cDNA are required for this reaction. Previous studies revealed that truncated forms of Rous sarcoma virus integrase containing two of the three protein domains can carry out integration reactions in vitro. Here, we describe the crystal structure at 2.5 A resolution of a fragment of the integrase of Rous sarcoma virus (residues 49-286) containing both the conserved catalytic domain and a modulatory DNA-binding domain (C domain). The catalytic domains form a symmetric dimer, but the C domains associate asymmetrically with each other and together adopt a canted conformation relative to the catalytic domains. A binding path for the viral cDNA is evident spanning both domain surfaces, allowing modeling of the larger integration complexes that are known to be active in vivo. The modeling suggests that formation of an integrase tetramer (a dimer of dimers) is necessary and sufficient for joining both viral cDNA ends at neighboring sites in the target DNA. The observed asymmetric arrangement of C domains suggests that they could form a rotationally symmetric tetramer that may be important for bridging integrase complexes at each cDNA end.
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Vírus do Sarcoma Aviário/enzimologia , Domínio Catalítico/fisiologia , Integrases/química , Integrases/metabolismo , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , Sequência de Aminoácidos , Substituição de Aminoácidos/genética , Sequência de Bases , Sítios de Ligação , Domínio Catalítico/genética , Cristalização , Cristalografia por Raios X , DNA/genética , DNA/metabolismo , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Dimerização , Desenho de Fármacos , Integrase de HIV/química , Inibidores de Integrase/química , Inibidores de Integrase/metabolismo , Integrases/genética , Modelos Moleculares , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida/genética , Fragmentos de Peptídeos/genética , Conformação ProteicaRESUMO
ABSTRACT Sugarcane mosaic is the most widespread virus disease affecting sugarcane production. We have established a collection of seven prominent sugarcane mosaic potyvirus (SCMV) strains currently causing disease in sugarcane throughout the world and originally found in sugarcane in the United States. This collection includes SCMV strains A, B, D, and E, and the sorghum mosaic virus (SrMV) strains SCH, SCI, and SCM. These viruses were propagated on Sorghum bicolor cv. Rio and purified. Cloned cDNAs representing 2.0 kb of the 3' termini, obtained after a reversetranscriptase-polymerase chain reaction (RT-PCR) on purified virions using an oligo(dT) primer and degenerate primers with sequences located in the NIb gene, have been sequenced for each of these strains. A comparative analysis of the deduced amino acid sequences in the NIb and coat protein genes and of the nucleotide sequences in the 3'-untranslated region, among these seven viruses and among six other members of the SCMV subgroup, confirms that there are at least four, but suggests that there are five, distinct viruses in the SCMV complex. Based on these seven new sequences and on the available sequence data for six other members of the SCMV subgroup, we have developed group-specific primers for use in RT-PCR-based restriction fragment length polymorphism analysis for rapid discrimination between strains of SCMV and SrMV. This is the first assay for differentiating strains of SCMV and SrMV that does not require interpretation of symptoms on differential hosts.
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Transgenic plants of grapefruit cv. Rio Red (Citrus paradisi Macf.) have been obtained by Agrobacterium tumefaciens-mediated gene transfer using seedling-derived epicotyl segments as explants and kanamycin as the selective agent. The transformation procedure includes a shoot elongation phase with a liquid medium overlay, which provides additional selection against non-transgenic shoots. Transformed shoots are invigorated and multiplied on a non-selective medium prior to grafting, thus assuring that plants can be recovered from transgenic shoots. We have constructed a binary vector, pBin34SGUS, with an intron-containing ß-glucuronidase gene (uidA) under the control of the Figwort mosaic virus 34S promoter. The 34S promoter efficiently drives uidA gene expression both in transient assays and in transgenic Rio Red leaf tissue, although at levels five- to sevenfold lower than the Cauliflower mosaic virus 35S promoter. An untranslatable coat protein gene (uncp) of the Citrus tristeza virus strain SY568 and the Galanthus nivalis agglutinin gene (gna) were inserted into pBin34SGUS and transgenic plants have been obtained. Stable integration of the uncp and gna genes was confirmed by Southern hybridization and gna gene expression was confirmed by Western blot analysis.
Assuntos
Álcool Desidrogenase/química , Isoenzimas/química , Conformação Proteica , Álcool Desidrogenase/isolamento & purificação , Álcool Desidrogenase/metabolismo , Sítios de Ligação , Clonagem Molecular , Simulação por Computador , Cristalografia por Raios X , Escherichia coli , Humanos , Isoenzimas/isolamento & purificação , Isoenzimas/metabolismo , Modelos Moleculares , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , SoftwareRESUMO
Open reading frame 79 of Bombyx mori nucleopolyhedrovirus (Bm79) is a conserved gene whose homologues have been identified in all 26 of the completely sequenced baculovirus genomes, including lepidopteran NPVs and GVs, hymenopteran NPVs, and a dipteran baculovirus. Northern blot analysis showed that the Bm79 transcript was about 850 nucleotides long and was initiated 12 h p.i. Temporal expression analysis revealed a 28-kDa protein, which was detected beginning 24 h p.i. using a polyclonal antibody against GST-Bm79 fusion protein. The 28-kDa protein was detected in the occlusion-derived virus envelope (ODV-E), but not in budded viruses. This observation was confirmed by observing ultrathin sections of polyhedra using immunoelectron microscopy. This demonstrated that the protein was present within the nuclei of cells. These results suggest that Bm79 is a functional gene that encodes a structural protein associated with the envelope of occlusion-derived virus (ODV).
Assuntos
Bombyx/virologia , Nucleopoliedrovírus/genética , Proteínas do Envelope Viral/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Northern Blotting , Western Blotting , Linhagem Celular , Núcleo Celular/metabolismo , Imunofluorescência , Expressão Gênica , Microscopia Imunoeletrônica , Dados de Sequência Molecular , Peso Molecular , Nucleopoliedrovírus/química , Fases de Leitura Aberta/genética , Alinhamento de Sequência , Fatores de Tempo , Proteínas do Envelope Viral/química , Proteínas do Envelope Viral/metabolismoRESUMO
Isolation of the terminal portions of genomic DNA cloned in bacterial artificial chromosomes (BACs) is an important step in map-based cloning, and several methods have been developed. Here, we present a new method based on double-restriction-enzyme digestion followed by anchored PCR. BAC DNA was digested with two enzymes: NotI and one of four enzymes (EcoRV, HpaI, StuI, or XmnI) that produce blunt termini. After dephosphorylation, these digestions were ligated to NotI- and EcoRV-digested pMSK, a new cloning vector developed in this work that is derived from pBluescript SK(+). PCR products representing the left- and right-terminal sequences of BAC inserts were obtained using a primer complementary to pMSK and a primer complementary to sequences in either the left arm or the right arm of the BAC vector pBeloBAC11. We have tested this method with 15 different BAC clones, and PCR products representing both the left- and right-terminal sequences have been obtained from all 15 BAC clones. This method is simple, fast, reproducible, and uses the same set of primers for any restriction enzyme used. With some modifications, it can also be used for isolating the terminal portions of genomic DNA cloned in yeast artificial chromosomes and P1-derived artificial chromosomes.
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Cromossomos , Genes Bacterianos , Passeio de Cromossomo , Clonagem Molecular , Enzimas de Restrição do DNA/metabolismo , Eletroforese em Gel de Ágar , Modelos GenéticosRESUMO
Free transplantation of subaxillary lateral thoracodorsal flap is recommended. The thoraco-dorsal artery and its accompanying veins may nourish and drain a large musculo-cutaneous flap. The calibres of both the artery and the veins are big enough to be anastomosed to the recipient vessels using the naked eye. The donor area can be closed by first intention suture without interfering with the functional recovery of the donor area. Both the functional and cosmetic results after free transplantations of this flap are good. In acute electrical or other deep burns, this flap can be used to prevent infection, to protect the deep vital structures, and to lessen the amputation rate. A better condition is created for functional recovery and facilitating reconstructive surgery. The muscular branch of the thoraco-dorsal artery was always present in our 50 patients whereas only 20 cases (40 per cent) had an independent cutaneous branch which nourishes the flap. We stress the point of preserving and using the muscular branch as the main blood vessel to be anastomosed for this free flap.