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1.
Artigo em Inglês | MEDLINE | ID: mdl-38808966

RESUMO

Fishmeal substitution with sustainable feed sources is highly essential towards sustainable production. This study aimed to investigate the effects of substituting fishmeal (FM) with Daphnia magna biomass meal (DBM) or zooplankton biomass meal (ZBM) on growth performance, liver and intestinal histology, gut bacterial abundance and stress tolerance of Nile tilapia, Oreochromis niloticus, fry. Nile tilapia fry (0.23 ± 0.04 g) were randomly assigned to five groups of three replicates. The control diet comprised 300 g/kg FM, and the FM was substituted with DBM or ZBM at levels of 25% and 50% (DBM-25, DBM-50, ZBM-25 and ZBM-50 respectively) in the other experimental diets. The experiment lasted 56 days in 1.5 m3 concrete tanks. The results revealed that weight gain and feed conversion ratio (FCR) significantly (p ≤ 0.035 and 0.025 respectively) improved with a polynomial response with a peak at 25% ZBM and a linear increase with DBM up to 50% of FM. Histometric indices of the distal intestine showed improvements (p ≤ 0.001) in villus height, villus width, crypt depth and muscle thickness of fish fed DBM or ZBM compared to the control. In the meantime, there were no histological abnormalities in the liver sections. The replacement of FM with DBM or ZBM could modulated gut bacterial abundance, including total bacterial count, Escherichia coli, Bacillus subtilis, and Lactobacillus sp. The fish-fed DBM or ZBM-containing diets had higher (p ≤ 0.05) tolerances to salinity stress than the control group. In conclusion, DBM or ZBM could replace FM up to 50% and 25%, respectively with improved fish growth performance, FCR, gut histology and tolerance to salinity stress.

2.
J Anim Physiol Anim Nutr (Berl) ; 108(3): 752-763, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38305567

RESUMO

The current study aimed to evaluate growth performance, digestive enzyme activities, antioxidant status, nonspecific immune response and intestinal histological status of red tilapia fed Daphnia meal (DM) as a substitute for fishmeal (FM). Hybrid red tilapia (Oreochromis mossambicus × Oreochromis aureus) fry (0.54 ± 0.05 g fish-1) was allocated in nylon haba cages (100 fry m-3) for 2 weeks as an acclimation period. The fish were divided into five groups (three replicates each). The experimental diets were prepared by replacing FM with DM at concentrations of 25%, 50%, 75% and 100% respectively. The results indicated that fish fed increasing levels of DM (50%-75%) experienced high growth performance, feed utilisation and protein content. The activities of digestive enzymes were significantly increased in all groups fed DM diets compared to the control. The antioxidant balance was improved by decreasing the level of malondialdehyde and increased the total antioxidant capacity, catalase, superoxide dismutase and glutathione reductase activities in the liver of fish fed DM. The nonspecific immune response, including lysozyme, alkaline phosphatase activities and total protein level improved significantly with increasing FM substitution levels by DM in a dose-dependent manner. Histometric analysis of the intestinal wall revealed an increase in the villus length, crypts depth and goblet cells number in groups fed DM meal up to 50% substitution level compared to other treatments. It may be concluded from results of this feeding trial that in the aquaculture of hybrid tilapia, FM may be substituted with up to 50% DM without compromising intestinal health, growth performance and immune status of the fish.


Assuntos
Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Antioxidantes , Dieta , Intestinos , Tilápia , Animais , Ração Animal/análise , Antioxidantes/metabolismo , Dieta/veterinária , Digestão/efeitos dos fármacos , Intestinos/efeitos dos fármacos , Tilápia/crescimento & desenvolvimento
3.
Toxics ; 10(10)2022 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-36287876

RESUMO

Sixty-seven sites of Hafafit pegmatite from the Southeastern Desert of Egypt were investigated radiometrically in the field using an in situ γ-ray spectrometer to determine eU, eTh, and K contents. The obtained results ranged from 0.4 to 6 ppm for eU with a mean value of 2.5 ppm, from 0.2 to 32 ppm for eTh with a mean value of 6.7 ppm, and from 0.7% to 5.4% for K with a mean value of 3.3%. Consequently, the radiological effects from these rocks were estimates by determination of the environmental parameters: gamma activity concentration index Iγ, external hazard index Hex, internal hazard index Hin, external absorbed dose rates in outdoor, and external absorbed dose rates in indoor air. The results obtained in this study showed that values U, Th, and K lie in the range of the acceptable world values. In addition, the calculated radiation hazard parameters (Iγ, Hex, and Hin) have values lower than the world values, while the calculated external absorbed dose rates (Dair) have values higher than the world and Egyptian permissible levels.

4.
Sci Rep ; 12(1): 13690, 2022 08 11.
Artigo em Inglês | MEDLINE | ID: mdl-35953507

RESUMO

Various studies have shown the importance of using different types of Zooplankton biomasses as an additional substance in the diet of fish. In addition, the drainage water of the fish cultures could be used in plant irrigation. In this study, biomasses of water flea Daphnia magna and Gammarus pulex collected and tested, for the first time, their effect against pathogenic microorganisms and on plant germination. The results showed significant antibacterial activity of D. magna and G. pulex against Staphylococcus aureus and Pseudomonas aeruginosa bacteria, as well as antifungal activity against Alternaria solani and Penicillium expansum, which gives the possibility to be used as biocontrol against these bacteria and plant pathogenic fungi. Furthermore, both animals showed positive activity in the germination rate of Vicia faba seed, reaching 83.0 ± 3.5 and 86.0 ± 3.8%, respectively. In conclusion, the biomasses of D. magna and G. pulex are promising and effective agents for their use in the medical field against some pathogenic microbes and as stimulators of plant growth.


Assuntos
Anfípodes , Daphnia , Animais , Bactérias , Fungos , Zooplâncton
5.
Egypt J Immunol ; 28(1): 12-22, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34147050

RESUMO

Accumulating evidence has indicated that immune regulatory cells are involved in the establishment of the anti-tumor activity, however; the role of regulatory B cells (B-regs) in breast cancer (BC) remains unclear. This study intended to assess the frequency of peripheral B-regs phenotypes in patients with BC, and to determine the relation between these phenotypes and the patient's clinicopathological characters. The expressions of the immune cell populations were analyzed by four-color flow cytometry in 40 naïve BC patients and 10 age-matched apparently healthy individuals as controls attending the department of Clinical Oncology and Nuclear Medicine at Assiut University Hospitals. The percentages of B-regs phenotypes CD19+IL10+ and CD19+CD24hiCD27+IL10+ were higher in BC patients than in the controls. The percentage of CD19+IL10+ B cells phenotype was significantly associated with the HER-2 expression levels, T, and N stages of BC. In conclusion, high percentage of B-regs phenotypes CD19+IL10+ and CD19+CD24hiCD27+IL10+ in BC patients indicates a possible role in immune suppression during the development of BC.


Assuntos
Linfócitos B Reguladores , Neoplasias da Mama , Antígenos CD19 , Egito/epidemiologia , Feminino , Citometria de Fluxo , Humanos , Fenótipo , Linfócitos T Reguladores
6.
PLoS One ; 16(1): e0244949, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33412574

RESUMO

BACKGROUND: Interleukin-18 (IL-18) and interferon-γ (IFN-γ) are cytokines of crucial role in inflammation and immune reactions. There is a growing evidence supporting important roles for IL-18 and IFN γ in tuberculosis (TB) infection and anti-tuberculosis immunity. OBJECTIVE: To evaluate the role of polymorphisms in IL-18-607 and -137 and INF-γ +874 in susceptibility to TB infection among Egyptian patients. METHODS: A case control study was conducted to investigate the polymorphism at IL-18-607, -137 and INF-γ+874 by sequence specific primer-polymerase chain reaction (SSP- PCR) in 105 patients with pulmonary and extra pulmonary tuberculosis and 106 controls. RESULTS: A significant protective effect against TB was found in homozygous CC genotype at IL-18 -137G/C, in addition to a 7-fold risk with GG and GC genotypes in the recessive model. Apart from a decreased risk with the AC genotype, no association was detected between the susceptibility to TB and different genotypes or alleles at the IL-18 -607A/C site. The homozygous AA genotype in INF-γ+874 showed a significant higher risk to TB than the homozygous TT or heterozygous AT genotypes with nearly a 2-fold risk of TB infection with the A allele. Regarding haplotype association, the GC haplotype was strongly associated with TB infection compared to other haplotypes. CONCLUSION: These findings suggest; for the first time in Egypt; a significant risk to TB infection with SNP at the IL-18-137G/C with no LD with SNP at the IL-18-607 site. The homozygous AA genotype in INF-γ+874 showed a significant higher risk to TB than the homozygous TT or heterozygous AT genotypes.


Assuntos
Predisposição Genética para Doença , Genótipo , Interferon gama/genética , Interleucina-18/genética , Tuberculose/genética , Adulto , Alelos , Antituberculosos/uso terapêutico , Estudos de Casos e Controles , Egito , Feminino , Frequência do Gene , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Tuberculose/tratamento farmacológico , Adulto Jovem
7.
Egypt J Immunol ; 27(1): 65-76, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33180389

RESUMO

This study was performed to determine the role of autophagy-related 16-like 1 (ATG16L1, rs2241880) and IL10 (rs1800872) polymorphisms in the susceptibility to and early prediction of breast cancer in Egyptians. The study included 50 breast cancer patients and 50 apparently healthy controls. The PCR-RFLP technique was used to detect ATG16L1 (rs2241880) and IL10 (rs1800872) genotypes. IL10 level was determined in serum by ELISA. The mean age of the patients was 54.2 years. Among the patients, 80% had no family history for breast cancer, 70% were postmenopausal, and 72% exhibited grade II tumors. Metastasis was detected in 18% of the patients, and 6% of the cases exhibited triple-negative receptor (TNR) status. In the ATG16LI (rs2241880) gene, the GG genotype frequency was significantly higher in patients than in controls (14% in patients versus 2% in controls, P =0.02), and no metastasis was observed in patients with the AA genotype (P=0.03). In the IL10 (rs1800872) gene, the A allele was observed in 30% of patients and 23% of controls, but the difference was insignificant (P=0.26). Also, the prevalence of the AA genotype was 8% in patients and 4% in controls (P=0.54). Serum IL10 levels were higher in patients than in controls (P < 0.001). Within the patient group, individuals with the IL10 (rs1800872) AA genotype showed significantly higher serum IL10 levels than those with the CC and CA+CC genotypes (P =0.03 and 0.04, respectively). In conclusion, in Egyptian breast cancer patients, the GG genotype of ATG16LI (rs2241880) may be associated with increased disease risk, and the AA genotype could be protective against metastasis.


Assuntos
Proteínas Relacionadas à Autofagia/genética , Neoplasias da Mama , Interleucina-10/genética , Neoplasias da Mama/genética , Estudos de Casos e Controles , Egito/epidemiologia , Feminino , Predisposição Genética para Doença , Genótipo , Humanos , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único
8.
Animals (Basel) ; 10(9)2020 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-32932710

RESUMO

A 60-day feeding trial was conducted to assess the effects of dietary supplementation with bile salts (BS), lutein (LTN), and their combination on growth, survival, carotenoid content, and antioxidant status of rabbitfish (Siganus rivulatus) larvae. Fish were fed four isonitrogenous (34.39% protein) and isoenergetic (20.57 kJ/g) diets supplemented with BS (0.15 g kg-1), LTN (0.1 g kg-1), BS+LTN (0.15 and 0.1 g kg-1, respectively), and a non-supplemented control diet. The results revealed that fish fed BS+LTN had the highest significant specific growth rate (4.37% day-1), feed efficiency (46.55%), and survival (97.78%). Lutein supplementation improved whole-body protein content, meanwhile, fish fed a BS-supplemented diet had a higher lipid content. The carotenoid deposition was significantly increased with LTN and BS+LTN in skin, muscle, and whole body compared to the control and BS treatment. All dietary supplementation of BS and LTN showed significant improvement in total antioxidant capacity, catalase, and glutathione peroxidase activities. Additionally, LTN alone or BS+LTN significantly reduced malondialdehyde levels by 5.30 and 29.91%, respectively compared to the control. BS supplementation modulated aminopeptidases activities, triglycerides, cholesterol, and increased the activity of pancreatic lipase. Therefore, it could be inferred that dietary supplementation with LTN in combination with BS could improve the growth performance, carotenoid deposition, antioxidant status, lipid digestion, and metabolism of S. rivulatus.

9.
Int J Rheum Dis ; 23(9): 1226-1232, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32783391

RESUMO

Disturbances in autophagy are known to be implicated in autoimmune disorders. Many studies have connected polymorphisms in autophagy-related gene 5 (ATG-5) to systemic lupus erythematosus (SLE). Our aim was the determination of the expression level of ATG-5, Beclin-1 and microtubule-associated protein-light chain 3 (LC-3) in Egyptian SLE patients to investigate the impact of disturbances in autophagy genes on the incidence and progression of the disease. Also, we investigated the incidence of single nucleotide polymorphism (SNP) rs573775 in ATG-5 gene among Egyptian SLE patients. Our results showed that the mean levels of Beclin-1, LC-3 and interleukin (IL)-10 transcripts were significantly higher in SLE patients compared to healthy controls. The previous transcripts were positively correlated with SLE Disease Activity Index (SLEDAI). Beclin-1 and LC-3 transcripts were negatively correlated to complement component 3 (C3) levels. Only LC-3 transcripts were negatively correlated to complement component 4 (C4). The rs573775 SNP of ATG-5 with the variant allele was significantly associated with disease susceptibility, conferring a higher risk of SLE development. This variant allele was more prevalent in patients below 30 years, patients with anemia and in patients with anti-double-stranded DNA (dsDNA), confirming the essential role of ATG-5 polymorphism in the susceptibility of Egyptian patients to SLE.


Assuntos
Proteína 5 Relacionada à Autofagia/genética , Lúpus Eritematoso Sistêmico/genética , Polimorfismo de Nucleotídeo Único , Adulto , Proteína 5 Relacionada à Autofagia/sangue , Proteína Beclina-1/sangue , Proteína Beclina-1/genética , Biomarcadores/sangue , Estudos de Casos e Controles , Complemento C3/análise , Complemento C4/análise , Egito , Feminino , Estudos de Associação Genética , Predisposição Genética para Doença , Humanos , Interleucina-10/sangue , Interleucina-10/genética , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/diagnóstico , Masculino , Proteínas Associadas aos Microtúbulos/sangue , Proteínas Associadas aos Microtúbulos/genética , Fenótipo , RNA Mensageiro/sangue , RNA Mensageiro/genética , Medição de Risco , Fatores de Risco
10.
Arch Dermatol Res ; 312(3): 207-212, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31728627

RESUMO

Diagnosis of onychomycosis requires microbiological studies, which are time-consuming. Dermoscopy is non invasive, easy and coastless method. To evaluate the diagnostic role of dermoscopy in onychomycosis and comparing its findings with microbiological results. Eighty patients with onychomycosis and 40 controls were studied for nail dermoscopic finding, and microbiological examinations in the form of microscopic examination by 20% KOH, Sabouraud dextrose agar (SDA), and HiCrome Candida Differential Agar. 72.5% of the patients were females. Most of the patient were presented with one finger (35%) and two fingers (35%). 85% of the patient were presented clinically with distal lateral subungual onychomycosis followed by total dystrophic onychomycosis (12.5%) and lastly with superficial white onychomycosis (2.5%). 52.5% and 75% of the patients were positive by direct microscopic examination with 20%KOH and SDA, respectively. Dermatophytes isolated from 7.5% of the patient, non-dermatophytes (Aspergillus) was isolated from 2.5%, and 65% had Candida by SDA. C. albicans was the commonest species (75%), followed by C. tropicalis (17.3%), and lastly C. krusei (7.7%). Dermoscopic examinations of patients showed nail spikes, longitudinal striations, and color changes in 75%, 82.5%, and 95%, respectively, with statistically significant P value (P < 0.001). There was significant difference regarding long striations and yellow coloration dermoscopic finding with positive KOH patients. All patients with positive culture showed nail spikes on dermoscopic examination. Dermoscopy is a rapid tool for diagnosis of onychomycosis. Longitudinal striations is the best diagnostic dermoscopic finding. Microbiological test are still needed for accurate and reliable diagnosis.


Assuntos
Aspergillus/fisiologia , Candida/fisiologia , Dermoscopia/métodos , Unhas/patologia , Onicomicose/diagnóstico , Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Unhas/microbiologia , Adulto Jovem
11.
Infect Drug Resist ; 12: 2703-2710, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31564919

RESUMO

PURPOSE: To determine the bacteriological pattern and antibiotic susceptibility of bacterial isolates causing neonatal sepsis in Qena University Hospitals and compare polymerase chain reaction (PCR) and blood culture results in a trial for rapid diagnosis. PATIENTS AND METHODS: Blood samples from 75 clinically suspected cases of neonatal sepsis were subjected to identification of bacteria and determination of their antibiotic sensitivity through blood culture, and rapid detection of 16S rRNA and the uidA gene (to confirm the presence of E. coli) by PCR from extracted bacterial DNA. RESULTS: Most patients were preterm (64%) and low birth weight (LBW) (68%). In total, 42.7% presented with early onset sepsis (EOS). LBW was significantly associated with EOS (P-value=0.03). Although the blood culture and PCR results were similar in EOS, the PCR results were significantly higher than those of blood culture in detecting bacteria (85.3% vs 68%, respectively, P-value=0.001). Blood culture showed 100% specificity. The most common pathogen was E. coli (86.2%) in EOS and Staphylococcus spp. (45.5%) in late-onset sepsis (LOS) (P-value=0.001 and 0.02, respectively). The most effective antibiotics against Gram-negative bacteria were ofloxacin, ciprofloxacin, imipenem, and amikacin, while vancomycin, oxacillin, and imipenem were the most effective antibiotics against Gram-positive bacteria. CONCLUSION: EOS was mainly caused by E. coli, while LOS was mainly caused by Staphylococcus spp. The 16S rRNA PCR showed higher sensitivity with rapid and accurate diagnosis. Blood culture is the most suitable method for antimicrobial sensitivity testing.

12.
Egypt J Immunol ; 26(1): 113-120, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31333001

RESUMO

Type 1 diabetes (T1D) is an autoimmune disease associated with multiple genetics and environmental factors. The aim of the study is to determine the frequency of HLA-B*08 and HLA-B*39 and its linkage disequilibrium with common risk haplotypes DR3-DQA1*05-DQB1*02, and DR4-DQA1-03-DQB1*0302 among T1D Egyptian infants. And assess different environmental factors as early exposure to cow's milk, exclusive breast feeding, mode of delivery and low birth weight. Sixty eight diabetic infants and 120 healthy controls were studied. HLA-DQB1, and DQA1 alleles were identified using homogeneous PCR and oligonucleotide hybridization assays. HLA-B*08 and HLA-B*39 genes were identified using multiplex PCR. The results showed that early exposure to cow's milk before 6 months carry a significant risk for T1D (16% in patients versus 6.6%in control group, P value=0.03). HLA-B*08 frequency was significantly higher among T1D infants than in control group (14.5% in diabetic infants versus 5%in control group, P value=0.024). DR3-DQA1*05-DQB1*02, and DR4-DQA1-03-DQB1*0302 were significantly higher in diabetic infants than controls (P value < 0.001 and 0.004 respectively). HLA-B*08 gene was found in (15.5%) of DR3-DQA1*05-DQB1*02 positive cases while in control group it was found in (13.5%) (P value=0.8). In conclusion, HLA-B*08 gene carry a risk for T1D in Egyptian infants, while DR3-DQA1*05-DQB1*02 haplotype lacks linkage disequilibrium with HLA-B*08 among T1D infants. Further studies are needed to determine which HLA-B gene is strongly linked to DR3-DQA1*05-DQB1*02 haplotype in T1D infants other than HLA-B*08 and HLA-B*18.


Assuntos
Diabetes Mellitus Tipo 1/genética , Antígenos HLA-B/genética , Cadeias alfa de HLA-DQ/genética , Cadeias beta de HLA-DQ/genética , Antígeno HLA-DR3/genética , Desequilíbrio de Ligação , Leite , Alelos , Animais , Estudos de Casos e Controles , Bovinos , Egito , Predisposição Genética para Doença , Haplótipos , Humanos , Lactente , Fatores de Risco
13.
Egypt J Immunol ; 26(1): 121-128, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31333002

RESUMO

Interleukin-35 (IL-35), is a recently identified cytokine that belongs to the IL-12 family, it is a potent anti-inflammatory and immunosuppressive cytokine which was first recognized to be produced by regulatory T cells (Tregs) cells, and recently was found to be produced by regulatory B cells (Bregs). The study aimed at determining whether plasma levels of IL-35 are associated with clinical characteristics of breast cancer (BC) patients. The study included 40 patients with breast cancer (BC), and 10 matched controls. The IL-35 cytokine was measured in plasma using ELISA. Results showed that plasma IL-35levels were significantly higher in BC than healthy controls (P˂ 0.05), and were significantly associated with BC grade 2 and HER-2 over expression level "3+", suggesting that plasma IL-35 levels may be associated with the development and progression of BC.


Assuntos
Neoplasias da Mama/sangue , Interleucinas/sangue , Linfócitos B Reguladores , Estudos de Casos e Controles , Feminino , Hospitais Universitários , Humanos , Linfócitos T Reguladores
14.
Mol Biol Rep ; 46(1): 1157-1165, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30628022

RESUMO

The immunomodulatory property of mesenchymal stem cells (MSCs) has been previously reported. Still it is unclear if this property can be affected by the cell origin and cell quality. Using primary MSCs expanded from bone marrow (BM-MSCs) and adipose tissue (AD-MSCs) of mice, we investigated whether the immunomodulatory property of MSCs varied with cell origin and cell quality (early- vs. late-passaged BM-MSCs). BM-MSCs (p1) and AD-MSCs (p1) had a typical spindle shape, but morphological changes were observed in late-passaged BM-MSCs (p6). A pathway-focused array showed that the expression of chemokine/cytokine genes varied with different cell origins and qualities. By co-culturing with spleen mononuclear cells (MNC) for 3 days, the expression of CD4 was suppressed by all types of MSCs. By contrast, the expression of CD8 was suppressed by BM-MSCs and increased by AD-MSCs. The expression ratio of CD206 to CD86 was at a comparable level after co-culture with AD-MSCs and BM-MSCs, but was lower with late-passaged BM-MSCs. AD-MSCs highly induced the release of IL6, IL-10 and TGF-ß in culture medium. Compared with early-passaged BM-MSCs (p1), late-passaged BM-MSCs (p6) released less TGF-ß. Our data suggests that the immunomodulatory properties of MSCs vary with cell origin and cell quality and that BM-MSCs of good quality are likely the optimal source of immunomodulation.


Assuntos
Imunomodulação/imunologia , Células-Tronco Mesenquimais/imunologia , Células-Tronco Mesenquimais/fisiologia , Tecido Adiposo/citologia , Animais , Células da Medula Óssea/citologia , Diferenciação Celular/fisiologia , Proliferação de Células , Células Cultivadas , Técnicas de Cocultura , Citocinas/metabolismo , Expressão Gênica/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL
15.
BMC Res Notes ; 10(1): 720, 2017 Dec 08.
Artigo em Inglês | MEDLINE | ID: mdl-29221488

RESUMO

OBJECTIVE: Recombinant adeno-associated virus (AAV)-based vectors are characterized by their robust and safe transgene delivery. The CRISPR/Cas9 and guide RNA (gRNA) system present a promising genome-editing platform, and a recent development of a shorter Cas9 enzyme from Staphylococcus aureus (SaCas9) allows generation of high titer single AAV vectors which carry both saCas9- and gRNA-expression cassettes. Here, we used two AAV-SaCas9 vectors with distinct GFP-targeted gRNA sequences and determined the impact of AAV-SaCas9-gRNA vector treatment in a single cell clone carrying a GFP-expression cassette. RESULTS: Our results showed comparable GFP knockout efficiencies (40-50%) upon a single low-dose infection. Three consecutive transductions of 25-fold higher doses of vectors showed 80% GFP knockout efficiency. To analyze the "AAV-SaCas9-resistant cell population", we sorted the residual GFP-positive cells and assessed their permissiveness to super-infection with two AAV-Cas9-GFP vectors. We found the sorted cells were significantly more resistant to the GFP knockout mediated by the same AAV vector, but not by the other GFP-targeted AAV vector. Our data therefore demonstrate highly efficient genome-editing by the AAV-SaCas9-gRNA vector system. Differential susceptibilities of single cell-derived cells to the AAV-SaCas9-gRNA-mediated genome editing may represent a formidable barrier to achieve 100% genome editing efficiency by this vector system.


Assuntos
Proteínas Associadas a CRISPR/genética , Sistemas CRISPR-Cas , Dependovirus , Endonucleases/genética , Sequenciamento do Exoma , Edição de Genes , Vetores Genéticos , RNA Guia de Cinetoplastídeos , Staphylococcus aureus/enzimologia , Proteínas de Bactérias , Proteína 9 Associada à CRISPR , Linhagem Celular , Suscetibilidade a Doenças , Proteínas de Fluorescência Verde , Células HEK293 , Humanos
16.
Urol Int ; 98(4): 411-417, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-27871076

RESUMO

OBJECTIVES: To assess the efficacy of adjunctive tamsulosin therapy in improving the success rate of laser-assisted semi-rigid ureteroscopy (URS) for removing proximal ureteral stones. PATIENTS AND METHODS: This prospective study included 165 patients with proximal ureteral stones ≥10 mm. The patients were randomly assigned to a tamsulosin group (Group I, n = 81) receiving tamsulosin 0.4 mg daily for 1 week pre-URS and a control group (Group II, n = 84) without tamsulosin therapy. Treatment consisted of URS using a semi-rigid ureteroscope (7.5 Fr), followed by intracorporeal holmium: YAG laser lithotripsy. The patients were followed up regularly for 8 weeks after URS. RESULTS: The operative time was 43.4 and 49.6 min in Groups I and II, respectively (p < 0.001). Scope to stone access rate was 93.8 and 82.1% in patients of Groups I and II, respectively (p = 0.022). The stone-free rate was significantly higher in Group I compared to Group II (74/81; 91.4% vs. 67/84; 79.8%; p = 0.035). The complication rate was significantly lower in Group I compared to Group II (17.3 vs. 38.1%, p = 0.003). Only minor complications were encountered and were managed conservatively. CONCLUSIONS: Tamsulosin therapy prior to semi-rigid URS improved ureteroscopic access to proximal ureteral stones, thus leading to an increased success rate and low morbidity.


Assuntos
Sulfonamidas/uso terapêutico , Cálculos Ureterais/tratamento farmacológico , Cálculos Ureterais/cirurgia , Ureteroscópios , Adulto , Terapia Combinada , Feminino , Humanos , Litotripsia a Laser , Masculino , Pessoa de Meia-Idade , Duração da Cirurgia , Probabilidade , Estudos Prospectivos , Tansulosina , Fatores de Tempo , Resultado do Tratamento , Ureteroscopia
17.
Arab J Urol ; 14(2): 94-100, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27489735

RESUMO

OBJECTIVE: To identify the criteria of phosphodiesterase type 5 inhibitor (PDE5i) users and to analyse the knowledge, attitude, and practices of PDE5i use amongst Saudi men. SUBJECTS AND METHODS: A web-based, cross-sectional survey was conducted in Saudi Arabia between January and April 2015. Sexually active adult men were interviewed using a website questionnaire designed by the authors. Descriptive statistics were used to analyse the data. RESULTS: In all, 1008 men participated in the survey with 378 (37.5%) reporting use of PDE5i. Of those using PDE5i, 144 (38.1%) reported erectile dysfunction and 234 (61.9%) reported normal erection (recreational users). We found several demographic features, including high education level, health field occupation, high income, smoking, and increased frequency of sexual intercourse amongst the PDE5i users. Most of the PDE5i users (92.3%) had knowledge about PDE5i and 84.1% of them bought it without medical prescription. The most commonly used PDE5i was tadalafil (46.1%) and most of the users (79.9%) reported improvement in their sexual activity after PDE5i usage. Amongst the recreational users, the main reasons for PDE5i usage were curiosity (38.5%) and improving self-confidence (25.6%). Of them, 69.2% reported benefits from PDE5i usage, mainly in the form of enhancement of erection (36.7%) and increasing erection duration (31.2%). CONCLUSION: PDE5i use appears to be frequent in Saudi Arabia. Most of the users had knowledge about PDE5i and claimed to get benefits from it, even if used as a recreational drug.

18.
Arch Virol ; 161(11): 3161-9, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27544760

RESUMO

The IL28B gene is associated with spontaneous or treatment-induced HCV viral clearance. However, the mechanism by which the IL28B single nucleotide polymorphism (SNP) affects the extra-hepatic HCV immune responses and its relationship to HCV pathogenesis have not been thoroughly investigated. To examine the mechanism by which IL28B affects HCV clearance. Forty Egyptian patients with chronic HCV infection receiving an Interferon/ribavirin treatment regimen were enrolled into this study. There were two groups: non-responders (NR; n = 20) and sustained virologic responders (SVR; n = 20). The initial plasma HCV viral loads prior to treatment and IL28B genotypes were determined by quantitative RT-PCR and sequencing, respectively. Liver biopsies were examined to determine the inflammatory score and the stage of fibrosis. Colonic regulatory T cell (Treg) frequency was estimated by immunohistochemistry. No significant association between IL28B genotypes and response to therapy was identified, despite an odds ratio of 3.4 to have the TT genotype in NR compared to SVR (95 % confidence interval 0.3-35.3, p = 0.3). Patients with the TT-IL28Brs12979860 genotype (unfavorable genotype) have significantly higher frequencies of colonic Treg compared to the CT (p = 0.04) and CC (p = 0.03) genotypes. The frequency of colonic Treg cells in HCV-infected patients had a strong association with the IL-28B genotype and may have a significant impact on HCV clearance.


Assuntos
Colo/imunologia , Interleucinas/genética , Mucosa Intestinal/imunologia , Polimorfismo de Nucleotídeo Único , Linfócitos T Reguladores/imunologia , Adulto , Antivirais/uso terapêutico , Estudos Transversais , Egito , Feminino , Hepatite C Crônica/tratamento farmacológico , Histocitoquímica , Humanos , Interferon-alfa/uso terapêutico , Interferons , Fígado/patologia , Masculino , Pessoa de Meia-Idade , Plasma/virologia , Ribavirina/uso terapêutico , Resultado do Tratamento , Carga Viral , Adulto Jovem
19.
J Med Microbiol ; 65(8): 703-712, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27166142

RESUMO

Extra-hepatic compartments might contribute to hepatitis C virus (HCV) persistence and extra-hepatic manifestations. Therefore, we investigated HCV infection in colonic tissue in patients with chronic hepatitis C (CHC) and its relationship with HCV pathogenesis. Colonic biopsies were collected from three groups with CHC infection: treatment naïve (TN; n=12), non-responders (NR; n=10) to anti-HCV therapy (pegylated interferon-α and ribavirin) and sustained virologic response (SVR; n=10) and from a fourth healthy control group (n=10). Liver biopsies were examined to assess inflammation and fibrosis. HCV infection and colonic T regulatory (Treg) frequency were detected by immunohistochemistry. HCV core and NS3 proteins were detected in B cells and macrophage/monocytes of 42 % and 25 % of TN and 50 % and 30 % of NR, respectively, but not in SVR or control group. The numbers of cells expressing HCV proteins were positively correlated with both HCV viral load and colonic Treg frequency. A significant negative correlation between HCV-expressing cells with both liver inflammation and fibrosis was identified. Our study provides evidence that HCV can infect B cells and macrophages of the colon. The correlations between HCV infection in colonic tissue and HCV viral load and liver pathology underline the significance of this extra-hepatic infection in HCV pathogenesis and response to therapy.


Assuntos
Colo/virologia , Hepacivirus/isolamento & purificação , Hepatite C Crônica/virologia , Adulto , Linfócitos B/virologia , Biópsia , Estudos Transversais , Feminino , Humanos , Imuno-Histoquímica , Fígado/virologia , Macrófagos/virologia , Masculino , Pessoa de Meia-Idade , Linfócitos T Reguladores/imunologia , Adulto Jovem
20.
Urol Int ; 97(3): 266-272, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27082108

RESUMO

INTRODUCTION: To evaluate the efficacy of adjunctive tamsulosin therapy after extracorporeal shockwave lithotripsy (SWL) for renal stones. MATERIALS AND METHODS: This prospective study was conducted on patients who underwent single-session SWL for solitary renal stone less than 20 mm. Post-SWL, patients were randomly divided into 2 groups; the tamsulosin group (TG), received a daily dose of tamsulosin 0.4 mg, for a maximum of 12 weeks, with post-SWL traditional analgesia and control group (CG), received the traditional analgesia alone. RESULTS: The study enrolled 271 patients and eventually only 249 (123 in TG and 126 in CG) completed the study. The overall stone free rate was 73.5%: 78.0% in TG and 69.0% in CG (p = 0.108). In TG, 8.1% of patients experienced at least one episode of acute renal/ureteral colic compared with 19.8% of controls (p = 0.008). The mean cumulative analgesia dosage per subject was 313 mg in TG and 346 mg in CG (p < 0.001). Overall, 14 patients developed steinstrasse and 6 of them (all in CG) needed urgent intervention (p = 0.031). CONCLUSIONS: Daily tamsulosin therapy does not enhance the clearance of stone fragments but decreases the pain episodes, analgesia dosage and need for adjuvant intervention after SWL for renal stones.


Assuntos
Antagonistas de Receptores Adrenérgicos alfa 1/uso terapêutico , Cálculos Renais/terapia , Litotripsia , Sulfonamidas/uso terapêutico , Adulto , Terapia Combinada , Feminino , Humanos , Cálculos Renais/tratamento farmacológico , Masculino , Estudos Prospectivos , Tansulosina
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