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1.
Int J Reprod Biomed ; 19(7): 607-618, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34458669

RESUMO

BACKGROUND: Coenzyme Q10 (CoQ10) and Lepidium sativum (LS) have therapeutic effects on infertility. OBJECTIVE: To evaluate the combined effects of LS and CoQ10 on reproductive function in adult male NMRI mice. MATERIALS AND METHODS: Eighty three-months-old male mice (35-40 gr) were divided into four groups (n = 10/each): control (treated with water), CoQ10-treated (200, 300, and 400 mg/kg/body weight), LS-treated (200, 400, 600 mg/kg/body weight), and co-treated (LS [600 mg/kg/body weight] + CoQ10 [200 mg/kg/body weight]) groups. Serum testosterone, luteinizing hormone, follicle-stimulating hormone, and gonadotropin realizing hormone (GnRH) levels were measured using ELISA method. The sperm quality was assessed using Sperm Class AnalyzerⓇ (SCA) CASA system and GnRH mRNA expression levels were evaluated by real-time polymerase chain reaction. RESULTS: The number of sniffing and following behavior was significantly higher in LS-treated (400 and 600 mg/ml/body weight) groups than the control group (p = 0.0007 and p = 0.0010, respectively). The number of mounting and coupling behaviors was significantly higher in the CoQ10 (300 and 400 mg/ml/body weight)-treated animals than the control group (p = 0.0170 and p = 0.0006, respectively). Co-treatment of CoQ10 (200 mg/ml/body weight) and LS (600 mg/ml/body weight) significantly increased all aspects of sexual behaviors as well as the levels of serum testosterone (p = 0.0011), luteinizing hormone (p = 0.0062), and follicle-stimulating hormone (p = 0.0001); sperm viability (p = 0.0300) and motility (p = 0.0010); and GnRH mRNA levels (p = 0.0016) compared to the control group. CONCLUSION: The coadministration of CoQ10 and LS significantly improves the activity of the hypothalamic-pituitary-gonadal axis and enhances the reproductive parameters in adult male mice.

2.
BMC Med Genomics ; 14(1): 37, 2021 02 02.
Artigo em Inglês | MEDLINE | ID: mdl-33530996

RESUMO

BACKGROUND: Autosomal recessive non-syndromic hearing loss (ARNSHL) is genetically and phenotypically heterogeneous with over 110 genes causally implicated in syndromic and non-syndromic hearing loss. Here, we investigate the genetic etiology of deafness in two GJB2 and GJB6 negative patients presenting with pre-lingual, progressive, severe hearing loss. METHODS: Targeted exome sequencing (TES) using Next Generation Illumina Sequencing was used to analyze the exonic and some other important genomic regions of 154 genes in the proband. Subsequently, the mutation found was confirmed by Sanger sequencing in other affected sibling and healthy family members. The possible impact of the reported mutation on the corresponding protein was also evaluated by using bioinformatics tools. Moreover, the affected patients underwent audiological and ophthalmic evaluations. RESULTS: TES identified a novel homozygous missense mutation c.251T>C (p.I84T) in exon 3 of PDZD7 gene. In addition, segregation and phenotype-genotype correlation analysis as well as in-silico evaluations confirmed the autosomal recessive inheritance pattern and disease-causing nature of mutation found. CONCLUSIONS: In overall, our finding could expand the pathogenic mutations spectrum and strengthens the clinical importance of the PDZD7 gene in ARNSHL patients. It can also aid to conduct genetic counseling, prenatal diagnosis and clinical management of these types of genetic disorders.


Assuntos
Surdez , Estudos de Associação Genética , Humanos , Irã (Geográfico) , Masculino
3.
Life Sci ; 264: 118719, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33159957

RESUMO

AIM: As a natural compound, docosahexaenoic acid (DHA) exerts anti-cancer and anti-angiogenesis functions through exosomes; however, little is known about the molecular mechanisms. MAIN METHODS: Breast cancer (BC) cells were treated with DHA (50 µM) and then tumor cell-derived exosomes (TDEs) were collected and characterized by electron microscopy, dynamic light scattering, and western blot analyses. By the time the cells were treated with DHA, RT-qPCR was used to investigate the expression of vascular endothelial growth factor (VEGF) and the selected pro- and anti-angiogenic microRNAs (miRNAs). The quantification of secreted VEGF protein was measured by enzyme-linked immunosorbent assay (ELISA). The effects of TDEs on endothelial cell angiogenesis were explored by transwell cell migration and in vitro vascular tube formation assays. KEY FINDINGS: DHA treatment caused a significant and time-dependent decrease in the expression and secretion of VEGF in/from BC cells. This also increased expression of anti-angiogenic miRNAs (i.e. miR-34a, miR-125b, miR-221, and miR-222) while decreased levels of pro-angiogenic miRNAs (i.e. miR-9, miR-17-5p, miR-19a, miR-126, miR-130a, miR-132, miR-296, and miR-378) in exosomes derived from DHA-treated BC cells, TDE (DHA+). While treatment with exosomes (100 µg/ml) obtained from untreated BC cells, TDE (DHA-), enhanced the expression of VEGF-A in human umbilical vein endothelial cells (HUVECs), incubation with DHA or TDE (DHA+) led to the significant decrease of VEGF-A transcript level in these cells. We indicated that the incubation with TDE (DHA+) could significantly decrease endothelial cell proliferation and migration and also the length and number of tubes made by HUVECs in comparison with endothelial cells incubated with exosomes obtained from untreated BC cells. SIGNIFICANCE: DHA alters angiogenesis by shifting the up-regulation of exosomal miRNA contents from pro-angiogenic to anti-angiogenic, resulting in the inhibition of endothelial cell angiogenesis. These data can help to figure out DHA's anti-cancer function, maybe its use in cancer therapy.


Assuntos
Neoplasias da Mama/patologia , Movimento Celular/efeitos dos fármacos , Ácidos Docosa-Hexaenoicos/farmacologia , Exossomos/metabolismo , Células Endoteliais da Veia Umbilical Humana/patologia , Neovascularização Fisiológica/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Endocitose/efeitos dos fármacos , Exossomos/efeitos dos fármacos , Exossomos/ultraestrutura , Feminino , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Fatores de Tempo , Fator A de Crescimento do Endotélio Vascular/metabolismo
4.
Biotechnol Appl Biochem ; 68(6): 1250-1256, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33012018

RESUMO

Breast cancer (BC) is one of the most common malignancies among women in the world. There is a global attempt to diagnose BC as early as possible. Long noncoding RNAs (lncRNAs) are emerging as novel targets and biomarkers for BC diagnosis and prognosis. Aberrant expression of lncRNAs is associated with BC development, making them a potential tumor marker for BC. To investigate this possibility, we determined the expression levels of Down syndrome cell adhesion molecule-antisense RNA-1 (DSCAM-AS1) and mitotically-associated long non-coding RNA (MANCR) lncRNAs in BC tissues. This case-control study included 50 paired tumor and adjacent nontumor tissues from female BC patients. The total RNA was isolated and the expression levels of MANCR and DSCAM-AS1 lncRNAs were assessed using quantitative real-time reverse transcription-PCR. Potential correlations between lncRNA levels and clinicopathological characteristics were also analyzed. DSCAM-AS1 and MANCR lncRNAs were significantly upregulated in BC tumor tissues compared with the adjacent nontumor tissues. We also found the significant upregulation of DSCAM-AS1 in advanced tumor-node-metastasis stage (TNM III) of BC tumor tissues. Furthermore, the expression of DSCAM-AS1 and MANCR in HER-2 positive patients was significantly higher than HER-2 negative affected individuals. Receiver operating characteristic curve analysis showed a satisfactory diagnostic efficacy (P value < 0.0001), which means that DSCAM-AS1 and MANCR lncRNAs can potentially serve as a biomarker. The present study might provide further approval for the clinical diagnostic significance of DSCAM-AS1 and MANCR lncRNAs that their high expressions were associated with aggressive clinical parameters of BC.


Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/metabolismo , RNA Longo não Codificante/metabolismo , Regulação para Cima , Biomarcadores Tumorais/genética , Neoplasias da Mama/diagnóstico , Estudos de Casos e Controles , Feminino , Humanos , Pessoa de Meia-Idade , RNA Longo não Codificante/genética
5.
Life Sci ; 248: 117466, 2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-32101760

RESUMO

AIMS: Nanoparticles (NPs)-based drugs have been recently introduced to improve the efficacy of current therapeutic strategies for the treatment of cancer; however, the molecular mechanisms by which a NP interacts with cellular systems still need to be delineated. Here, we utilize the autophagic potential of TiO2 NPs for improving chemotherapeutic effects of 5-fluorouracil (5-FU) in human AGS gastric cells. MATERIALS AND METHODS: Cell growth and viability were determined by trypan blue exclusion test and MTT assay, respectively. Vesicular organelles formation was evaluated by acridine orange staining of cells. Cell cycle and apoptosis were monitored by flow cytometry. Reactive oxygen species (ROS) level were measured by DCHF-DA staining. Autophagy was examined by q-PCR and western blotting. Molecular docking was used for studying NP interaction with autophagic proteins. KEY FINDINGS: TiO2 NPs increase ROS production, impair lysosomal function and subsequently block autophagy flux in AGS cells. In addition, the autophagy blockade induced by non-toxic concentrations of TiO2 NPs (1 µg/ml) can promote cytotoxic and apoptotic effects of 5-FU in AGS cells. SIGNIFICANCE: These results confirm the beneficial effects of TiO2 NPs in combination with chemotherapy in in vitro model of gastric cancer, which may pave the way to develop a possible solution to circumvent chemoresistance in cancer.


Assuntos
Antimetabólitos Antineoplásicos/farmacologia , Autofagia/efeitos dos fármacos , Fluoruracila/farmacologia , Regulação Neoplásica da Expressão Gênica , Nanopartículas/química , Titânio/farmacologia , Antimetabólitos Antineoplásicos/química , Apoptose/efeitos dos fármacos , Apoptose/genética , Autofagia/genética , Proteína 5 Relacionada à Autofagia/genética , Proteína 5 Relacionada à Autofagia/metabolismo , Proteína 7 Relacionada à Autofagia/genética , Proteína 7 Relacionada à Autofagia/metabolismo , Proteína Beclina-1/genética , Proteína Beclina-1/metabolismo , Ciclo Celular/efeitos dos fármacos , Ciclo Celular/genética , Linhagem Celular Tumoral , Proliferação de Células , Vesículas Citoplasmáticas/efeitos dos fármacos , Vesículas Citoplasmáticas/metabolismo , Sinergismo Farmacológico , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Fluoruracila/química , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/metabolismo , Mucosa Gástrica/patologia , Humanos , Lisossomos/efeitos dos fármacos , Lisossomos/metabolismo , Proteínas Associadas aos Microtúbulos/antagonistas & inibidores , Proteínas Associadas aos Microtúbulos/química , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Simulação de Acoplamento Molecular , Nanopartículas/ultraestrutura , Conformação Proteica , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Espécies Reativas de Oxigênio/agonistas , Espécies Reativas de Oxigênio/metabolismo , Proteína Sequestossoma-1/antagonistas & inibidores , Proteína Sequestossoma-1/genética , Proteína Sequestossoma-1/metabolismo , Titânio/química , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
6.
Infect Disord Drug Targets ; 20(2): 160-166, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-30706828

RESUMO

BACKGROUND: Salmonellosis is a major food-borne disease worldwide. The increasing prevalence of antimicrobial resistance among food-borne pathogens such as Salmonella spp. is concerning. OBJECTIVE: The main objective of this study is to identify class 1 integron genes and to determine antibiotic resistance patterns among Salmonella isolates from children with diarrhea. METHODS: A total of 30 Salmonella isolates were recovered from children with diarrhea. The isolates were characterized for antimicrobial susceptibility and screened for the presence of class 1 integron genes (i.e. intI1, sulI1, and qacEΔ1). RESULTS: The most prevalent serotype was Enteritidis 36.7%, followed by Paratyphi C (30%), and Typhimurium (16.7%). The highest rates of antibiotic resistance were obtained for nalidixic acid (53.3%), followed by streptomycin (40%), and tetracycline (36.7%). Regarding class 1 integrons, 36.7%, 26.7%, and 33.3% of the isolates carried intI1, SulI, and qacEΔ1, respectively, most of which (81.8%) were multidrug-resistant (MDR). Statistical analysis revealed that the presence of class 1 integron was significantly associated with resistance to streptomycin and tetracycline (p = 0.042). However, there was no association between class 1 integron and other antibiotics used in this study (p > 0.05). CONCLUSION: The high frequency of integron class 1 gene in MDR Salmonella strains indicates that these mobile genetic elements are versatile among different Salmonella serotypes, and associated with reduced susceptibility to many antimicrobials.


Assuntos
Antibacterianos/farmacologia , Diarreia/microbiologia , Farmacorresistência Bacteriana Múltipla/genética , Integrons/genética , Salmonella/efeitos dos fármacos , Salmonella/genética , Adolescente , Criança , Pré-Escolar , Estudos Transversais , DNA Bacteriano/genética , Feminino , Humanos , Lactente , Irã (Geográfico) , Masculino , Testes de Sensibilidade Microbiana , Infecções por Salmonella/microbiologia , Sorogrupo
7.
Iran J Basic Med Sci ; 22(9): 1004-1009, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31807243

RESUMO

Objectives: Prevalence of high-fat food consumption, such as fast foods is one of the major causes of hypercholesterolemia, which can lead to cardiovascular diseases. 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) and cytochrome P450 7A1 (CYP7A1) are two key genes in cholesterol metabolism. Use of probiotics in the diet is a promising approach for modulation of serum lipid. To confirm the modulation of serum lipids by probiotics, in this study, we have examined the efficacy of Lactobacillus paracasei TD3 in improving blood cholesterol levels. Materials and Methods: 21 male Wistar rats were divided into three groups randomly (n=7). G1: negative control with normal diet, G2: positive control with high-fat diet, G3T: test group with high-fat diet plus supplementation with L. paracasei TD3 (1010 CFU). In the 21st day, the rats were anesthetized using chloroform and then sacrificed. Blood samples were collected to analyze lipid panel parameters and hepatic enzymes by the auto-analyzer system. Adipose tissue samples were analyzed using real-time PCR for HMGCR and CYP7A1 genes expression. Results: Consumption of L. paracasei TD3 could reduce serum cholesterol levels significantly (P<0.05); whereas, there was no significant difference between experimental groups for triglycerides, LDL, and HDL levels. Aspartate aminotransferase (AST) and Alanine aminotransferase (ALT) enzymes were significantly decreased in the probiotic group. Furthermore, expression of HMGCR and CYP7A1 genes was dramatically declined in the probiotic group. There was no significant change in either uric acid or urea between the control and treated groups. Conclusion: Introduction of L. paracasei TD3 in rat's diet can modulate serum cholesterol levels.

8.
BMC Med Genet ; 20(1): 147, 2019 08 29.
Artigo em Inglês | MEDLINE | ID: mdl-31464584

RESUMO

BACKGROUND: The SLC29A3 gene, encoding a nucleoside transporter protein, is found in intracellular membranes. Based on the literatures, mutations in this gene cause a wide range of clinical manifestations including H syndrome, pigmented hypertrichosis with insulin dependent diabetes, Faisalabad histiocytosis, and dysosteosclerosis. However, all these disorders with their different names and terminologies are actually the same entity termed H syndrome. CASE PRESENTATION: We report four GJB2 and GJB6 negative deaf patients from two Iranian related families who present the associated symptoms of SLC29A3-disorder. Whole Exome Sequencing (WES) using Next Generation Illumina Sequencing was used to enrich all exons of protein-coding genes as well as some other important genomic regions in one of studied patients. A novel homozygous frame-shift mutation c.307-308delTT (p.Phe103fs) in exon 3 of SLC29A3 gene was identified in a 35 years old man with profound hearing loss, camptodactyly, rheumatoid arthritis and delayed puberty without any skin changes, short stature and insulin dependent diabetes mellitus. The mutation found was also confirmed by Sanger sequencing in other studied patients and their healthy parents. In compared to proband, however the clinical manifestations of these patients were different, indicating variable expressivity of mutant SLC29A3 gene as well as possible involvement of other modifier genes. CONCLUSION: The present study uncovered a rare novel homozygous frame-shift mutation c.307-308delTT in SLC29A3 gene of four related patients with various manifestation of SLC29A3-disorder. Such studies can help to conduct genetic counseling and subsequently, prenatal diagnosis more accurately for individuals at the high risk of these types of genetic disorders.


Assuntos
Contratura/genética , Mutação da Fase de Leitura , Predisposição Genética para Doença/genética , Perda Auditiva Neurossensorial/genética , Histiocitose/genética , Homozigoto , Proteínas de Transporte de Nucleosídeos/genética , Adulto , Sequência de Bases , Conexina 26 , Conexina 30/genética , Conexinas/genética , Diabetes Mellitus Tipo 1/genética , Éxons , Feminino , Estudos de Associação Genética , Humanos , Irã (Geográfico) , Masculino , Pessoa de Meia-Idade , Linhagem
9.
EXCLI J ; 17: 1054-1068, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30564083

RESUMO

Dengue virus is a mosquito-borne pathogen that causes dengue diseases. All four serotypes of dengue virus are infectious for humans. Therefore, an efficacious dengue vaccine should be tetravalent to provide protection against all types of virus. The goal of this study was to design a new tetravalent recombinant protein from envelope protein of dengue viruses to induce virus-neutralizing antibodies against all four serotypes in mice. A chimeric protein was designed from domain III of envelope protein of all serotypes of dengue virus. Four domain III fragments were linked together by alpha helix making linkers. The final sequence of the designed protein was analyzed in silico and the coding gene sequence was deduced by reverse translation. After cloning and expression of the recombinant protein (ED3-tetravalent protein), identity of the purified protein was confirmed using a pan-dengue specific monoclonal antibody in Western blotting. Then, the immunogenicity of the purified protein was studied in mice using antibody titration. The efficacy of induced antibodies in neutralization of the virus was studies by FRNT method. Furthermore, the induction of cellular immunity was studied by measurement of cytokines using ELISA method and measurement of lymphocyte proliferation using MTT assay. The ED3-tetravalent protein was able to enhance neutralizing immunogenic response against all four dengue serotypes; in similar way to that of tetravalent formulation of four individual domain III-based polypeptides. It is suggested that the ED3-tetravalent fusion protein can induce broadly neutralizing antibody responses against all four serotypes of dengue virus in mice.

10.
Microb Pathog ; 123: 183-189, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30017942

RESUMO

Colorectal cancer is the third most common cause of cancer-related death in the world which genetic and environmental agents are responsible for cancer. When cells detach from the tumor and invade surrounding tissues, the tumor is malignant and may form secondary tumors at other locations in a process called metastasis. Probiotics are the largest group of inhabitation bacteria in the colon. Gut microbiota has a central role in prevented the risk colon cancer. Probiotics are beneficial microorganisms, like Lactic acid bacteria and Lactobacilli bacteria which are using in the dairy industry. Probiotics nisin are having the most important category of safe usage. In this study LS180, SW48, HT29 and Caco2 was cultured and treated with different dose of nisin. Cell proliferation was assayed with MTT. The expression of CEA, CEAM6 and MMP2F genes was analyzed with Real-time PCR. Protein expression of CEA was evacuated with ELISA. Our result was shown that the 40-50 IU/mL nisin could suppress proliferation of LS180. Cell proliferation of SW48, HT29, Caco2 cells was decreased in 250-350 IU/mL concentration of nisin. The gene expression of CEA, CEAM6, MMP2F was significantly down-regulated with nisin treatment (p < 0.001, p < 0.01). Also, after cells treated with nisin, CEA protein expression was down regulated (p < 0.01). In conclusion, nisin could suppressed metastatic process via down-regulation of CEA, CEAM6, MMP2F, MMP9F genes. We suggested the new treatment strategies beyond Probiotics, which play a role in the prevention local tumor invasion, metastasis and recurrence.


Assuntos
Antibacterianos/farmacologia , Bacteriocinas/farmacologia , Neoplasias Colorretais/tratamento farmacológico , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Nisina/farmacologia , Células CACO-2/efeitos dos fármacos , Antígeno Carcinoembrionário/genética , Antígeno Carcinoembrionário/metabolismo , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Regulação para Baixo , Células HT29/efeitos dos fármacos , Humanos , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Nisina/administração & dosagem , Probióticos/farmacologia , Proteínas/genética , Proteínas/metabolismo
11.
Appl Microbiol Biotechnol ; 102(7): 2977-2996, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29470620

RESUMO

Dengue viruses are emerging mosquito-borne pathogens belonging to Flaviviridae family which are transmitted to humans via the bites of infected mosquitoes Aedes aegypti and Aedes albopictus. Because of the wide distribution of these mosquito vectors, more than 2.5 billion people are approximately at risk of dengue infection. Dengue viruses cause dengue fever and severe life-threatening illnesses as well as dengue hemorrhagic fever and dengue shock syndrome. All four serotypes of dengue virus can cause dengue diseases, but the manifestations are nearly different depending on type of the virus in consequent infections. Infection by any serotype creates life-long immunity against the corresponding serotype and temporary immunity to the others. This transient immunity declines after a while (6 months to 2 years) and is not protective against other serotypes, even may enhance the severity of a secondary heterotypic infection with a different serotype through a phenomenon known as antibody-depended enhancement (ADE). Although, it can be one of the possible explanations for more severe dengue diseases in individuals infected with a different serotype after primary infection. The envelope protein (E protein) of dengue virus is responsible for a wide range of biological activities, including binding to host cell receptors and fusion to and entry into host cells. The E protein, and especially its domain III (EDIII), stimulates host immunity responses by inducing protective and neutralizing antibodies. Therefore, the dengue E protein is an important antigen for vaccine development and diagnostic purposes. Here, we have provided a comprehensive review of dengue disease, vaccine design challenges, and various approaches in dengue vaccine development with emphasizing on newly developed envelope domain III-based dengue vaccine candidates.


Assuntos
Vacinas contra Dengue/imunologia , Vírus da Dengue/imunologia , Domínios Proteicos/imunologia , Proteínas do Envelope Viral/imunologia , Dengue/patologia , Dengue/prevenção & controle , Vacinas contra Dengue/genética , Humanos
12.
Artigo em Inglês | MEDLINE | ID: mdl-28713777

RESUMO

Cysteine/histidine-dependent amidohydrolase/peptidase (CHAP) and amidase are known as catalytic domains of the bacteriophage-derived endolysin LysK and were previously reported to show lytic activity against methicillin-resistant Staphylococcus aureus (MRSA). In the current study, the in silico design and analysis of chimeric CHAP-amidase model was applied to enhance the stability and solubility of protein, which was achieved through improving the properties of primary, secondary and tertiary structures. The coding gene sequence of the chimeric CHAP-amidase was synthesized and subcloned into the pET-22(+) expression vector, and the recombinant protein was expressed in E. coli BL21 (DE3) strain. Subsequent affinity-based purification yielded ~12 mg soluble protein per liter of E. coli culture. Statistical analysis indicated that concentrations of ≥1 µg/mL of the purified protein have significant antibacterial activity against S. aureus MRSA252 cells. The engineered chimeric CHAP-amidase exhibited 3.2 log reduction of MRSA252 cell counts at the concentration of 10 µg/mL. A synergistic interaction between CHAP-amidase and vancomycin was detected by using checkerboard assay and calculating the fractional inhibitory concentration (FIC) index. This synergistic effect was shown by 8-fold reduction in the minimum inhibitory concentration of vancomycin. The chimeric CHAP-amidase displayed strong antibacterial activity against S. aureus, S. epidermidis, and enterococcus. However, it did not indicate any significant antibacterial activity against E. coli and Lactococcus lactis. Taken together, these findings suggest that our chimeric CHAP-amidase might represent potential to be used for the development of efficient antibacterial therapies targeting MRSA and certain Gram-positive bacteria.


Assuntos
Amidoidrolases/farmacologia , Antibacterianos/farmacologia , Endopeptidases/química , Endopeptidases/farmacologia , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Proteínas Virais/farmacologia , Amidoidrolases/química , Amidoidrolases/genética , Sequência de Aminoácidos , Bacteriófagos/química , Bacteriófagos/enzimologia , Bacteriófagos/genética , Sequência de Bases , Linhagem Celular , Clonagem Molecular , Simulação por Computador , DNA Bacteriano , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Combinação de Medicamentos , Sinergismo Farmacológico , Endopeptidases/genética , Enterococcus/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Regulação Bacteriana da Expressão Gênica , Genes Bacterianos , Concentração de Íons de Hidrogênio , Lactococcus lactis/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , Simulação de Acoplamento Molecular , Estabilidade Proteica , Estrutura Terciária de Proteína , Proteínas Recombinantes/genética , Análise de Sequência , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus epidermidis , Temperatura , Vancomicina/farmacologia , Proteínas Virais/química , Proteínas Virais/genética
13.
Clin Exp Vaccine Res ; 5(1): 41-9, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26866023

RESUMO

PURPOSE: Dengue virus infection is now a global problem. Currently, there is no licensed vaccine or proven antiviral treatment against this virus. All four serotypes (1-4) of dengue virus can infect human. An effective dengue vaccine should be tetravalent to induce protective immune responses against all four serotypes. Most of dengue vaccine candidates are monovalent, or in the form of physically mixed multivalent formulations. Recently envelope protein domain III of virus is considered as a vaccine candidate, which plays critical roles in the most important viral activities. Development of a tetravalent protein subunit vaccine is very important for equal induction of immune system and prevention of unbalanced immunity. Here, we have presented and used a rational approach to design a tetravalent dengue vaccine candidate. MATERIALS AND METHODS: We designed a multi domain antigen by fusing four consensus domain III sequences together with appropriate hydrophobic linkers and used several types of bioinformatics software and neural networks to predict structural and immunological properties of the designed tetravalent antigen. RESULTS: We designed a tetravalent protein (EDIIIF) based on domain III of dengue virus envelope protein. According to the results of the bioinformatics analysis, the constructed models for EDIIIF protein were structurally stable and potentially immunogenic. CONCLUSION: The designed tetravalent protein can be considered as a potential dengue vaccine candidate. The presented approach can be used for rational design and in silico evaluation of chimeric dengue vaccine candidates.

14.
Iran J Basic Med Sci ; 17(11): 836-43, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25691924

RESUMO

OBJECTIVES: Production of a recombinant and immunogenic antigen using dengue virus type-3 envelope protein is a key point in dengue vaccine development and diagnostic researches. The goals of this study were providing a recombinant protein from dengue virus type-3 envelope protein and evaluation of its immunogenicity in mice. MATERIALS AND METHODS: Multiple amino acid sequences of different isolates of dengue virus type-3, corresponding to the envelope protein domain III, were achieved from GenBank. Clustal V alignment tool was used to provide a consensus amino acid sequence. Nucleotide sequence of the coding gene was optimized using "Optimizer". The origami (DE3) strain of Escherichia coli was used as the host in order to express the protein. A commercial affinity chromatography method was used to purify the recombinant protein. Immunogenicity of the recombinant protein was evaluated in mice using ELISA, MTT and cytokine assays. RESULTS: A consensus amino acid sequence corresponding to the most important region of dengue virus type-3 envelope protein (domain III) was provided. A high concentration (≥ 20 mg/L culture medium) of soluble recombinant antigen (EDIII3) was achieved. Immunized mice developed specific antibody responses against EDIII3 protein. The splenocytes from EDIII3-immunized mice showed a high proliferation rate in comparison with the negative control. In addition, the concentrations of two measured cytokines (IFN-γ and IL-4) were increased markedly in immunized mice. CONCLUSION: The results showed that the expressed recombinant EDIII3 protein is an immunogenic antigen and can be applied to induce specific immune responses against dengue virus type-3.

15.
Arch Iran Med ; 15(5): 298-302, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22519379

RESUMO

BACKGROUND: The aim of this study was to assess the possibility of a primary end-to-end pharyngoesophageal anastomosis after standard tumor resection of the cervical esophagus by acute flexion of the neck. METHODS: A total of 34 consecutive patients with primary cervical esophageal cancer, none having received prior radio- or chemotherapy, were treated by two methods based on intraoperative findings. In 18 patients, reconstruction after esophageal resection was carried out by the standard gastric pull-through technique (control group). In 16 patients, acute flexion of the neck after tumor resection allowed for reconstruction by primary end-to-end pharyngoesophagostomy (experimental group). RESULTS: There was no operative mortality in either group. The mean operative time for the experimental group was about 50 minutes less compared to the control group. Self-limited postoperative anastomotic leakage in the neck was twice as common in the experimental group. Postoperative dysphagia was about three times as common in the experimental group [5 patients (31%)] compared to the control group [2 patients (11%)]. CONCLUSION: In selected cases, segmental resection of primary cervical esophageal cancers reconstructed by end-to-end pharyngoesophagostomy is technically feasible by bending the neck acutely forward during anastomosis and maintaining it in the flexed position during a postoperative period of about 7 days. The advantages are reduced scope and duration of the operation. The downside is doubling of the frequency of postoperative cervical leakage.


Assuntos
Anastomose Cirúrgica , Neoplasias Esofágicas , Humanos , Pescoço/cirurgia , Estômago
16.
JOP ; 11(6): 617-9, 2010 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-21068498

RESUMO

CONTEXT: Cystic lymphangiomas originate as benign masses which occur mostly in children especially in the head and neck region and/or the groin. Although abdominal lymphangiomas are rare, they are most commonly reported in adults. In addition, pancreatic involvement is rare. Lymphatic malformation with blockage of the lymphatic flow is the most common etiology leading to the formation of lymphangiomas. Cystic lymphangiomas should always be included in the differential diagnosis of abdominal masses which present with mass effect signs and symptoms. Due to its rarity, it forms a diagnostic and therapeutic challenge for the clinician. CASE REPORT: We herein report the case of a 43-year-old man with a cystic lymphangioma detected in the head of the pancreas and describe the surgical procedure utilized as the therapeutic medium. CONCLUSION: To remove this mass, we utilized a modified approach to a classic pancreaticoduodenectomy. This technique involved resection of the head of the pancreas while preserving the upper 2nd portion of the duodenum and the ampulla of Vater. The result of our 30-month follow-up of this patient has been very satisfactory with no complications.


Assuntos
Linfangioma Cístico/diagnóstico , Linfangioma Cístico/cirurgia , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/cirurgia , Adulto , Humanos , Linfangioma Cístico/patologia , Masculino , Modelos Biológicos , Neoplasias Pancreáticas/patologia , Radiografia Abdominal , Tomografia Computadorizada por Raios X
17.
Head Neck ; 25(9): 772-7, 2003 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-12953314

RESUMO

BACKGROUND: Wide resection of oropharyngeal malignancies implicates the risk of velopharyngeal insufficiency, which can cause nasal regurgitation and hypernasality. A meticulous reconstruction is necessary to avoid impairment and handicap in deglutition and speech. In the classic reconstructive techniques for large oropharyngeal defects, functional outcome only regards deglutition. We also focus on nasality, because hypernasality often occurs as a consequence in this type of reconstruction. METHODS: In four patients, the surgical defect is closed with a free radial forearm flap sutured to the posterior side of the hard palate, thus imitating a caudally based pharyngeal flap. Speech is assessed by an independent speech pathologist, perceptually and acoustically. Deglutition is evaluated by a questionnaire and videofluoroscopy. RESULTS: All patients had normal food intake. They did not report alterations in speech quality or verbal communication. Perceptual evaluation of articulation, voice, and nasality was optimal. Objective measurements with acoustical analysis and nasality scores confirmed the excellent functional outcome. Videofluoroscopy showed an unimpaired bolus transport with a complete velopharyngeal closure and optimal oral and pharyngeal clearance times. CONCLUSIONS: This meticulous reconstructive technique ensures an excellent functional outcome. The absence of nasality, in particular, proves the value of this refinement. The technique allows wide surgical margins and complete velopharyngeal closure.


Assuntos
Neoplasias Orofaríngeas/cirurgia , Retalhos Cirúrgicos , Insuficiência Velofaríngea/cirurgia , Deglutição , Feminino , Humanos , Masculino , Neoplasias Orofaríngeas/complicações , Neoplasias Orofaríngeas/fisiopatologia , Projetos Piloto , Inteligibilidade da Fala , Resultado do Tratamento , Insuficiência Velofaríngea/etiologia , Insuficiência Velofaríngea/fisiopatologia
18.
Dysphagia ; 18(2): 78-84, 2003.
Artigo em Inglês | MEDLINE | ID: mdl-12825900

RESUMO

Our study compares deglutition between a group who had undergone total esophagopharyngolaryngectomy and a group who had esophagectomy and partial pharyngectomy with preserved larynx, after reconstruction of the upper digestive tract with pedicled colon interposition. In four patients the laryngeal structures could be preserved (three caustic burns and one proximal esophageal tumor). Six patients underwent a total laryngopharyngectomy for large pharyngeal tumors. Swallowing was assessed by a questionnaire, clinical examination, and videofluoroscopy. All patients had normal intake of semisolid foods and fluids. All patients but three experienced some feeling of "narrowing" of the tract: four at the level of the hypopharynx, two at the oropharyngeal level, one at the oral level. In the laryngectomy group, solid food caused some degree of delayed swallowing in three patients. Dumping occurred in one case out of the nonlaryngectomy group. On clinical examination a tense motility in all laryngectomy patients appeared, food remnants in five and repeated swallowing movements in four. The videofluoroscopy confirmed repeated swallowing movements and presence of residual food in the oral cavity. Temporal stagnation occurred at the anastomosis site in all patients and in two patients at a place of colon redundancy. Colon interposition is a reliable reconstruction and gives the possibility of a good functional outcome. Although preservation of the larynx facilitates swallowing even in this reconstructive procedure, it may be better to perform a total laryngopharyngectomy and colon interposition in oncological cases where the pharyngeal remnant is borderline for primary closure.


Assuntos
Queimaduras Químicas/fisiopatologia , Queimaduras Químicas/cirurgia , Colo/fisiopatologia , Colo/transplante , Transtornos de Deglutição/fisiopatologia , Transtornos de Deglutição/cirurgia , Neoplasias Esofágicas/fisiopatologia , Neoplasias Esofágicas/cirurgia , Esofagectomia , Neoplasias Hipofaríngeas/fisiopatologia , Neoplasias Hipofaríngeas/cirurgia , Laringectomia , Avaliação de Resultados em Cuidados de Saúde , Faringectomia , Recuperação de Função Fisiológica/fisiologia , Adulto , Queimaduras Químicas/complicações , Transtornos de Deglutição/etiologia , Neoplasias Esofágicas/complicações , Feminino , Humanos , Neoplasias Hipofaríngeas/complicações , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos
19.
Eur Arch Otorhinolaryngol ; 260(1): 7-8, 2003 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-12520348

RESUMO

We present a patient treated by a total pharyngolaryngoesophagectomy and postoperative radiotherapy for a hypopharyngeal T4N2bM0 squamous cell carcinoma. The upper digestive tract was reconstructed with a pedicled left colon interposition through the posterior mediastinum. A voice prosthesis was placed 9 months after the initial treatment, following measurement of the tracheo-neopharyngeal wall thickness by sonography. Fifteen months after the total pharyngolaryngectomy, the patient remains free of recurrent disease and has successfully resumed speaking with the voice prosthesis.


Assuntos
Carcinoma de Células Escamosas/radioterapia , Carcinoma de Células Escamosas/cirurgia , Colo/transplante , Neoplasias Hipofaríngeas/radioterapia , Neoplasias Hipofaríngeas/cirurgia , Laringectomia/métodos , Laringe Artificial , Faringectomia/métodos , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Qualidade de Vida , Retalhos Cirúrgicos
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