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1.
SAGE Open Med ; 9: 20503121211049931, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34659762

RESUMO

Introduction: Critically ill COVID-19 patients are at increased risk of thrombosis with an enhanced risk of bleeding. We aimed to explore the role of anti-factor Xa levels in optimizing the high-intensity anticoagulation's safety and efficacy and finding possible associations between D-dimer levels, cytokine storm markers, and COVID-19-induced coagulopathy or thrombophilia. Methods: Retrospective cohort study conducted on 69 critically ill COVID-19 patients who received three regimens of higher intensity anticoagulation. Results: Seventeen patients (24.6%) received high-dose enoxaparin prophylaxis, 29 patients (42%) received therapeutic doses of enoxaparin, and 23 patients (33.3%) were on therapeutic unfractionated heparin infusion. Fewer than one-third of the whole cohort (n = 22; 31.8%) achieved the target range of anti-factor Xa. The patients were divided into three subgroups based on anti-factor Xa target status within each anticoagulation regimen; when compared, the only association observed among them was for interleukin-6 levels, which were significantly higher in both the "above the expected range" and "below the expected range" groups compared with the "within the expected range" group (p = 0.009). Major bleeding episodes occurred in 14 (20.3%) patients and were non-significantly more frequent in the "below the expected anti-factor Xa range group" (p = 0.415). Seven patients (10.1%) developed thrombosis. The majority of patients had anti-factor Xa levels below the expected ranges (four patients, 57.1%). Conclusion: Conventional anti-factor Xa ranges may not be appropriate as a predictive surrogate for bleeding in critically ill COVID-19. The clinical decision to initiate therapeutic anticoagulation preemptively may be individualized according to thrombosis and bleeding risks. Cytokine storm markers, namely, interleukin-6, may play a role in COVID-19-induced coagulopathy or thrombophilia.

2.
Molecules ; 26(13)2021 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-34206357

RESUMO

In the current work, a simple, economical, accurate, and precise HPLC method with UV detection was developed to quantify Favipiravir (FVIR) in spiked human plasma using acyclovir (ACVR) as an internal standard in the COVID-19 pandemic time. Both FVIR and ACVR were well separated and resolved on the C18 column using the mobile phase blend of methanol:acetonitrile:20 mM phosphate buffer (pH 3.1) in an isocratic mode flow rate of 1 mL/min with a proportion of 30:10:60 %, v/v/v. The detector wavelength was set at 242 nm. Maximum recovery of FVIR and ACVR from plasma was obtained with dichloromethane (DCM) as extracting solvent. The calibration curve was found to be linear in the range of 3.1-60.0 µg/mL with regression coefficient (r2) = 0.9976. However, with acceptable r2, the calibration data's heteroscedasticity was observed, which was further reduced using weighted linear regression with weighting factor 1/x. Finally, the method was validated concerning sensitivity, accuracy (Inter and Intraday's % RE and RSD were 0.28, 0.65 and 1.00, 0.12 respectively), precision, recovery (89.99%, 89.09%, and 90.81% for LQC, MQC, and HQC, respectively), stability (% RSD for 30-day were 3.04 and 1.71 for LQC and HQC, respectively at -20 °C), and carry-over US-FDA guidance for Bioanalytical Method Validation for researchers in the COVID-19 pandemic crisis. Furthermore, there was no significant difference for selectivity when evaluated at LLOQ concentration of 3 µg/mL of FVIR and relative to the blank.


Assuntos
Amidas/análise , Amidas/sangue , Antivirais/análise , Antivirais/sangue , Bioensaio/métodos , COVID-19/tratamento farmacológico , Cromatografia Líquida de Alta Pressão/métodos , Extração Líquido-Líquido/métodos , Pirazinas/análise , Pirazinas/sangue , Aciclovir/análise , Aciclovir/sangue , COVID-19/sangue , Calibragem , Estabilidade de Medicamentos , Congelamento , Humanos , Padrões de Referência , Reprodutibilidade dos Testes , Solventes/química
3.
Int J Mol Sci ; 22(8)2021 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-33923922

RESUMO

Doxorubicin increases endothelial permeability, hence increasing cardiomyocytes' exposure to doxorubicin (DOX) and exposing myocytes to more immediate damage. Reactive oxygen species are major effector molecules of doxorubicin's activity. Mangiferin (MGN) is a xanthone derivative that consists of C-glucosylxanthone with additional antioxidant properties. This particular study assessed the effects of MGN on DOX-induced cytotoxicity in human umbilical vein endothelial cells' (HUVECs') signaling networks. Mechanistically, MGN dramatically elevated Nrf2 expression at both the messenger RNA and protein levels through the upregulation of the PI3K/AKT pathway, leading to an increase in Nrf2-downstream genes. Cell apoptosis was assessed with a caspase-3 activity assay, transferase-mediated dUTP-fluorescein nick end labeling (TUNEL) staining was performed to assess DNA fragmentation, and protein expression was determined by Western blot analysis. DOX markedly increased the generation of reactive oxygen species, PARP, caspase-3, and TUNEL-positive cell numbers, but reduced the expression of Bcl-2 and antioxidants' intracellular concentrations. These were effectively antagonized with MGN (20 µM), which led to HUVECs being protected against DOX-induced apoptosis, partly through the PI3K/AKT-mediated NRF2/HO-1 signaling pathway, which could theoretically protect the vessels from severe DOX toxicity.


Assuntos
Doxorrubicina/farmacologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Xantonas/farmacologia , Antioxidantes/metabolismo , Apoptose/efeitos dos fármacos , Western Blotting , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Fragmentação do DNA/efeitos dos fármacos , Imunofluorescência , Humanos , Marcação In Situ das Extremidades Cortadas , Fator 2 Relacionado a NF-E2/genética , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética
4.
Chemosphere ; 260: 127551, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32683013

RESUMO

Semi-arid countries continue to face water scarcity, especially with the current global climatic changes. This scarcity has continuously increased over the last five decades in countries like Egypt, Syria, Libya and Jordan, where the agriculture sector consumes more than 85% of the country's water resources. The problem of water scarcity in Egypt is further challenged by high levels of urbanization, increasing industrial uses, and the high cost of advanced treatment processes. These challenges lead to the utilization of untreated or poorly treated wastewater for irrigation of agricultural crop fields. Thus, the current study proposes the use of an eco-friendly technology consisting of a constructed wetland planted with Typha latifolia and Cyperus papyrus supported with zeolite substrate for water purification, to curb this challenge. The results showed that, the removal efficiency of COD, BOD, TSS, and ammonia were 68.5%, 71%, 70%, and 82.3%, respectively by Typha latifolia bed. On the other hand, the removal efficiency of COD, BOD, TSS and ammonia were 85.5%, 86.2%, 83.9% and 92.3% respectively by Cyperus papyrus bed. As a result, bacteriological parameters were reduced to 99.9%, and complete removal of Salmonella sp was achieved during three days by Cyperus papyrus. Box-Behnken design was utilized to optimize independent factors, including contact time (24-72h) and initial concentration of metals (15-45 mg L-1) and their responses. The removal efficiency of Cu and Zn were 72% and 84%, respectively of the optimum reaction time (72 h), with 16 plant stems and an initial metal concentration of 15 mg L-1.


Assuntos
Metais Pesados/análise , Eliminação de Resíduos Líquidos/métodos , Áreas Alagadas , Agricultura , Biodegradação Ambiental , Cyperus , Egito , Enterobacteriaceae , Microbioma Gastrointestinal , Typhaceae , Águas Residuárias/análise , Purificação da Água
5.
Saudi J Med Med Sci ; 8(2): 140-145, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32587496

RESUMO

Background: Despite its importance, no study from the Middle East has assessed the neonatal pain knowledge of health-care providers in neonatal intensive care units (NICUs). Objectives: To evaluate the knowledge and attitudes of pediatricians and nurses in Saudi Arabia toward procedural pain assessment and its management in neonates. Materials and Methods: This questionnaire-based, cross-sectional study was conducted between March and June 2015 at three NICUs in the Eastern Province of Saudi Arabia, namely, King Fahd Hospital of the University, Al Khobar; Security Forces Hospital, Dammam; and King Fahd Medical Military Complex, Dhahran. All pediatricians/neonatologists and nurses in these units were included, and a previously validated questionnaire was distributed and collected by an assigned clinician. The questionnaire elicited pain-related knowledge of seven procedures using a 10-point Likert scale. Results: A total of 150 questionnaires were distributed, from which 107 complete responses were obtained (nurses: 81 [75.7%]; doctors: 26 [24.3%]). The overall knowledge scores were high (mean ± standard deviation = 77.3% ±11.6%). The mean pain rankings of doctors (7.2) were higher than those of nurses (6.5) for all procedures. Nurses reported more frequent use of analgesia than doctors (15.4% vs. 11.5%). Doctors often did not use comfort measures for any procedures, except during procedures on term to 28-day-old neonates. The use of pain tools was reported by only 12 (11%) clinicians. Conclusion: This study found that despite adequate knowledge about pain perception by neonates, the use of analgesia for procedural pain relief is low among doctors in the NICUs of the Eastern Province of Saudi Arabia. In addition, there is underutilization of pain assessment tools, thereby indicating the need to address this knowledge-practice gap.

6.
Vet Med Sci ; 6(3): 477-484, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32233074

RESUMO

BACKGROUND: The frequent outbreaks of Newcastle disease virus (NDV) in Iraq pose a constant threat to commercial poultry, despite the introduction of routine vaccination programmes. Several factors, particularly stress factors and coinfections, might play a role in increasing NDV outbreaks in poultry species. OBJECTIVES: The current study was aimed to characterize an NDV isolate from an outbreak in North Baghdad, Iraq. METHODS: Clinical pathogenicity of the isolate was determined experimentally in chickens. In vitro studies included cytopathological examination, as well as molecular and phylogenetic analyses. RESULTS: Based on the clinical studies and pathogenicity indices (mean death time and intracerebral and intravenous pathogenicity indices), the isolate was characterized as velogenic (highly virulent). Reverse transcriptase polymerase chain reaction targeting the partial fusion protein gene of the NDV genome confirmed the detection. Partial sequencing of the hypervariable region of the fusion gene identified the presence of an avirulent (lentogenic) fusion protein motif (GRQGRL). Phylogenetic analysis of the new isolate along with previously known regional isolates revealed that the new isolate was related to genotype II strains. Additionally, sequence analysis indicated a distinct genetic lineage of the new isolate, which was related to some of the lineages identified in previous outbreaks in the Middle East. CONCLUSION: The current study offers essential information on the epidemiology, characteristics and diagnosis of NDV for disease control in Iraq. The isolate was found to belong to genotype II and possess an avirulent fusion protein motif.


Assuntos
Galinhas , Doença de Newcastle/patologia , Vírus da Doença de Newcastle/isolamento & purificação , Vírus da Doença de Newcastle/patogenicidade , Doenças das Aves Domésticas/patologia , Animais , Genoma Viral , Genótipo , Iraque , Doença de Newcastle/virologia , Vírus da Doença de Newcastle/genética , Doenças das Aves Domésticas/virologia
7.
Water Environ Res ; 92(7): 1080-1088, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32012380

RESUMO

Human exposure to fluoride in drink water at a level above 1.5 mg/L causes a lot of health problems. The present study was carried out to assess the fluoride biosorption capability of Padina sp. alga as biosorbent material using conventional one-factor-at-a-time (OFAT) and Box-Behnken design to optimize the process. By OFAT, fluoride uptake was significantly affected by pH, time, fluoride concentration, and biosorbent dose (p-value < 0.05) and the highest fluoride biosorption (85.95%) was recorded at pH 7, time 60 min, fluoride concentration 2 g/L, and an adsorbent dose 30 g/L. Based on the Box-Behnken design, the quadratic model was developed to correlate the variables to the response. By Analysis of Variance (ANOVA), the model was statistically significant (p-value < 0.05). It was suggested that optimal values of pH, time, fluoride concentration, and biosorbent dose were 5, 90 min, 3 mg/L, and 30 g/L, respectively, and fluoride removal at these optimum conditions was 94.57%. For application, Padina sp. was removed from 67.79% to 78.78% of fluoride in collected groundwater samples. It was concluded that the Padina sp. could be used as eco-friendly biosorbent for fluoride and response surface methodology was more applicable and effective in optimization to obtain the highest removal efficiency of fluoride by Padina sp. PRACTITIONER POINTS: The fluoride biosorption was studied using Padina sp. alga as an eco-friendly biosorbent. By one-factor-at-a-time (OFAT), the highest fluoride biosorption rate was 85.95%. Response surface methodology was suggested that maximum Fluoride removal at optimum condition was 94.57%. Statistical response surface methodology was more applicable, effective in optimization and considered the interaction between factors.


Assuntos
Água Subterrânea , Poluentes Químicos da Água , Adsorção , Fluoretos , Concentração de Íons de Hidrogênio
8.
Am Heart J ; 220: 237-245, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31884246

RESUMO

RATIONALE: An estimated 15% of saphenous vein grafts (SVGs) occlude in the first year after coronary artery bypass grafting (CABG) despite aspirin therapy. Graft occlusion can result in symptoms, myocardial infarction, and death. SVG occlusion is primarily caused by atherothrombosis, in which platelet activation plays a pivotal role. Evidence regarding the effect of stronger platelet inhibition on SVG patency after CABG is limited. The main objective of the POPular CABG trial is to determine whether dual antiplatelet therapy with aspirin plus ticagrelor improves SVG patency when compared to aspirin alone. STUDY: The POPular CABG is a randomized, double-blind, placebo-controlled, multicenter trial investigating the effect of adding ticagrelor to standard aspirin therapy on the rate of SVG occlusion. A total of 500 patients undergoing CABG with ≥ 1 SVG are randomized to ticagrelor or placebo. The primary end point is SVG occlusion rate, assessed with coronary computed tomography angiography at 1 year. Secondary end points are stenoses and occlusions in both SVGs and arterial grafts and SVG failure at 1 year, defined as a composite of SVG occlusion on coronary computed tomography angiography or coronary angiography, SVG revascularization, myocardial infarction in the territory supplied by an SVG, or sudden death. Safety end points are bleeding events at 30 days and 1 year. CONCLUSION: The POPular CABG trial investigates whether adding ticagrelor to standard aspirin after CABG reduces the rate of SVG occlusion at 1 year.


Assuntos
Aspirina/uso terapêutico , Ponte de Artéria Coronária , Oclusão de Enxerto Vascular/tratamento farmacológico , Inibidores da Agregação Plaquetária/farmacologia , Veia Safena/transplante , Ticagrelor/farmacologia , Idoso , Angiografia por Tomografia Computadorizada , Angiografia Coronária/métodos , Morte Súbita Cardíaca/etiologia , Método Duplo-Cego , Quimioterapia Combinada , Oclusão de Enxerto Vascular/diagnóstico por imagem , Oclusão de Enxerto Vascular/epidemiologia , Humanos , Infarto do Miocárdio/etiologia , Placebos/farmacologia , Projetos de Pesquisa , Tamanho da Amostra , Grau de Desobstrução Vascular/efeitos dos fármacos
9.
Andrologia ; 51(10): e13400, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31489691

RESUMO

The aim of this study was to evaluate the relationship between the protamine ratio (P1/P2), DNA fragmentation of spermatozoa and protamine deficiency. Patients were grouped into fertile (G1; n = 151) and sub-fertile (G2; n = 121). DNA fragmentation in spermatozoa was analysed by a TUNEL assay (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling), and the protamination was determined by CMA3 staining, while Western blot was used to measure protamine P1 and P2. While sperm DNA fragmentation (SDF) and protamine ratio were significantly elevated in G2 compared with G1 (12.31 ± 7.01% vs. 17.5 ± 9.5%; p = .001) and (0.91 ± 0.43 vs. 0.75 ± 0.42; p = .003); respectively, the CMA3 positive showed no difference at all between G1 and G2. In G1, the CMA3 positive correlated negatively with the P1/P2 ratio and SDF (r = -.586, r = -.297; p = .001 respectively). In contrast, the protamine ratio correlated positively with SDF (r = .356; p = .001). In G2, no correlation was observed between CMA3 positive, SDF and the P1/P2 ratio but the P1/P2 ratio showed a positive correlation with SDF (r = .479; p = .001). In conclusion, the spermatozoa DNA deterioration was closely associated with abnormal protamination but showed an association with the protamine ratio, more than with CMA3 positive. Therefore, for the evaluation of DNA damage in spermatozoa, the P1/P2 ratio might act as an additional biomarker.


Assuntos
Infertilidade Masculina/diagnóstico , Protaminas/análise , Análise do Sêmen/métodos , Espermatozoides/metabolismo , Adulto , Biomarcadores/análise , Biomarcadores/metabolismo , Fragmentação do DNA , Humanos , Marcação In Situ das Extremidades Cortadas , Infertilidade Masculina/terapia , Masculino , Pessoa de Meia-Idade , Protaminas/metabolismo , Injeções de Esperma Intracitoplásmicas
10.
BMC Health Serv Res ; 19(1): 375, 2019 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-31196189

RESUMO

BACKGROUND: The levels of knowledge, attitude and practice among primary physicians concerning both diabetic retinopathy screening and treatment of sight threatening diabetic retinopathy have been studied by different groups, such as medical students, pharmacists, Primary Health Care staff and opticians. In some studies, the levels were very high, while in others it was noted to be less than desired. AIM: This study's intent is to estimate and improve level of Knowledge (K), Attitude (A) and Practice (P) among non-ophthalmic health care providers regarding eye management of diabetes and barriers that people with diabetes face in Saudi Arabia. METHODS: This cross-sectional survey targeted medical doctors (except ophthalmologists) working at private sector institutions in Riyadh. They were interviewed using closed-ended questions for knowledge (8), attitude (5), practice (5), and reasons for their current KAP status comprised of 8 questions. The level of Knowledge was assessed as good if its score was (> 50%); positive attitude (> 50%) and excellent practice (> 75%) were estimated and associated to the risk factors. RESULTS: Out of the 355 participants that were interviewed, the percentages of good knowledge, positive attitude and excellent practice concerning diabetic retinopathy (DR)were 193 [54.3% (95% CI 49.2-59.5)], 111 [31.3% (95% CI 26.4-36.1)], and 145 [40.8% (95% CI 35.7-46.0) participants, respectively. Gender, place of work and type of doctor were not significantly associated with the level of KAP. Salient reasons for low KAP status included a busy schedule (54.6%), less resources (75.2%), inadequate periodic training in eye care (69%), and absence of retinal evaluation training (49.6%). CONCLUSIONS: Improving KAP level is urgently needed. Addressing underlying causes of low KAP could enhance eye care of people with diabetes. Additionally, training for primary health care providers for early detection of DR and timely management of sight threatening diabetic retinopathy (STDR) is necessary.


Assuntos
Diabetes Mellitus/fisiopatologia , Retinopatia Diabética/diagnóstico , Pessoal de Saúde/estatística & dados numéricos , Programas de Rastreamento , Setor Privado , Estudos Transversais , Retinopatia Diabética/fisiopatologia , Conhecimentos, Atitudes e Prática em Saúde , Humanos , Encaminhamento e Consulta , Arábia Saudita
11.
Reprod Biol ; 19(1): 6-13, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30876814

RESUMO

The proper transition of histones to protamine during spermiogenesis is critical for male fertility. This study aimed to quantify the levels of histones and protamines mRNA in sperms of infertile couples and their possible effect on the sperm's quality and chromatin integrity. Spermatozoa from 53 normal and 75 patients were enrolled in this study. Histones and Protamine mRNAs were extracted, reverse-transcribed and applied to real-time quantitative PCR. Chromomycin A3 staining was used to assess protamination and chromatin integrity, and Eosin-Nigrosine-Test and HOS-Test was used to evaluate the sperm's vitality and membrane integrity respectively. Levels of histones H2A and H2B mRNA in patient's sperms were significantly (p < 0.01) higher than that of normal (31.22 ± 2.91, 30.03 ± 2.05 vs. 25.62 ± 1.98, 27.23 ± 3.04, respectively). Protamine PRM2 mRNA in patient's sperms (20.55 ± 2.01) was significantly lower than in normal (21.73 ± 2.64, p < 0.01). The PRM1/PRM2 and H2A/H2B mRNAs ratios were significantly higher (p < 0.01) in patients than normal (1.02 ± 0.10, 1.04 ± 0.07 vs. 0.98 ± 0.06, 0.94 ± 0.08 respectively). Also, the sperm's nuclear histones to protamines transcripts ((H2A + H2B)/(PRM1 + PRM2)) ratios of patients (1.49 ± 0.16) was significantly higher (p < 0.01) than that of normal (1.25 ± 0.15). Histone/protamine transcripts [((H2A + H2B)/(PRM1 + PRM2)) mRNAs ratios] were negatively correlated (p < 0.05) with sperm's count, total count, motility, progressive motility, normal morphology, membrane integrity and positively with chromatin decondensation. The data suggests that histones/protamines mRNAs ratios are important for a sperm's quality and therefore could be used as predictors for male infertility. Also, validation study may be required to confirm the study conclusion.


Assuntos
Histonas/metabolismo , Infertilidade Masculina , Protaminas/metabolismo , RNA Mensageiro/metabolismo , Espermatozoides/fisiologia , Adulto , Estudos de Casos e Controles , Sobrevivência Celular , Cromatina , Regulação da Expressão Gênica , Humanos , Masculino , Análise do Sêmen , Motilidade Espermática
12.
J Pharm Bioallied Sci ; 11(1): 49-59, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30906140

RESUMO

Context: Diet and beverages are thought to have notable effects on drugs. Recently, this relationship has received significant consideration. Aims: To develop and validate a simple, rapid, and sensitive method for the determination of glimepiride in rat serum. This will be performed using high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS). Potential pharmacokinetic interactions between glimepiride and the soft drink, Vimto, will also be investigated in the serum of experimental rats. Materials and Methods: HPLC-MS/MS was constructed and clarithromycin was used as an internal standard. Results: The method was validated in terms of linearity, precision, accuracy, stability, and system suitability parameters. The method was found to be satisfactory and suitable for the determination of glimepiride. The precision of glimepiride was high (coefficient of variation, CV% <15%), the accuracy over all 3 days of validation was within the accepted criteria. Glimepiride peak serum concentration (C max) was 126.01 ng/mL and was reached within 1 h (T max) of administration. Mean area under curve (AUC) was 964.70 ng/mL and was reached within 24 h of administration. The Vimto soft drink significantly (P < 0.050) reduced glimepiride peak serum concentration to 57.87 ng/mL and was reached within 2 h of administration. AUC was significantly reduced to 335.04 ng*h/mL (P < 0.050). Conclusion: Glimepiride pharmacokinetic parameters such as C max and AUC were significantly affected by the Vimto soft drink. Therefore, this study developed a simple, rapid, and sensitive method for validation and determination of the effects of soft drinks on drugs using the LC-MS/MS method.

13.
Hum Fertil (Camb) ; 22(2): 104-110, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28969455

RESUMO

This study was conducted to evaluate the relation between cigarette smoking, semen quality and ratios of protamine mRNAs in smokers and non-smokers. Spermatozoa from 123 men and 64 smokers and 59 non-smokers whose female partners attended an assisted reproduction and andrology laboratory were evaluated. Protamine mRNA was extracted from purified sperm, reverse-transcribed and subjected to real-time quantitative PCR using specific primer pairs for protamine 1 (PRM1) and protamine 2 (PRM2). The main outcomes showed that PRM1 mRNA levels in smokers were significantly lower (p = 0.05) than that of non-smokers. Additionally, PRM2 mRNA levels in smokers were significantly lower (p = 0.001) than that of non-smokers. PRM1/PRM2 mRNA ratios in non-smokers samples show significant differences (p = 0.001) compared with those in smokers. PRM1/PRM2 mRNA ratios were negatively and significantly correlated (p = 0.001) with semen volume, sperm count and normal sperm morphology. We concluded that sperm quality and sperm protamine mRNAs were negatively affected by smoking, and these data will serve as new evidence for the hazardous effect of smoking on male fertility. Additionally, protamine transcripts ratios may serve as a marker for male fertility.


Assuntos
Fumar Cigarros , Regulação da Expressão Gênica/efeitos dos fármacos , Protaminas/metabolismo , Espermatozoides/metabolismo , Humanos , Infertilidade Masculina , Masculino , Protaminas/genética , RNA Mensageiro , Análise do Sêmen
14.
Reprod Biomed Online ; 37(5): 581-589, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30366840

RESUMO

RESEARCH QUESTION: Does regular smoking affect semen quality and the levels of DNA methylation in mature human spermatozoa? DESIGN: Spermatozoa from 109 men were evaluated (55 smokers and 54 non-smokers). DNA was extracted from purified spermatozoa, and DNA methylation was quantified by enzyme-linked immunosorbent assay (ELISA). RESULTS: Global DNA methylation of non-smokers is significantly lower (P < 0.001) than that of smokers (4.85 ±â€¯2.72 and 7.08 ±â€¯1.77 ng/µl, respectively). Moreover, the mean global DNA methylation levels were significantly correlated (r = 0.22;P = 0.02) with non-condensed chromatin in the spermatozoa. Levels of non-condensed chromatin were significantly higher (P < 0.001) in smokers (29.75 ±â€¯9.38%) compared with non-smokers (20.96 ±â€¯11.31%). Furthermore, global sperm DNA methylation was negatively correlated with high significance (P < 0.010) with sperm: count (r = -0.27), motility (r = -0.30) and vitality (r = -0.26). CONCLUSION: Smoking interferes with DNA methylation. Also, DNA methylation is significantly correlated with sperm parameters and sperm non-condensed chromatin. These data emphasize another detrimental effect of smoking on male fertility. DNA methylation may, therefore, be considered as a fertility marker in men.


Assuntos
Metilação de DNA , Infertilidade Masculina/etiologia , Fumar/efeitos adversos , Espermatozoides/efeitos dos fármacos , Adulto , Ensaio de Imunoadsorção Enzimática , Humanos , Masculino , Análise do Sêmen
15.
Cell Host Microbe ; 23(5): 583-593.e8, 2018 05 09.
Artigo em Inglês | MEDLINE | ID: mdl-29746831

RESUMO

Helicobacter pylori (Hp) vacuolating cytotoxin (VacA) is a bacterial exotoxin that enters host cells and induces mitochondrial dysfunction. However, the extent to which VacA-dependent mitochondrial perturbations affect overall cellular metabolism is poorly understood. We report that VacA perturbations in mitochondria are linked to alterations in cellular amino acid homeostasis, which results in the inhibition of mammalian target of rapamycin complex 1 (mTORC1) and subsequent autophagy. mTORC1, which regulates cellular metabolism during nutrient stress, is inhibited during Hp infection by a VacA-dependent mechanism. This VacA-dependent inhibition of mTORC1 signaling is linked to the dissociation of mTORC1 from the lysosomal surface and results in activation of cellular autophagy through the Unc 51-like kinase 1 (Ulk1) complex. VacA intoxication results in reduced cellular amino acids, and bolstering amino acid pools prevents VacA-mediated mTORC1 inhibition. Overall, these studies support a model that Hp modulate host cell metabolism through the action of VacA at mitochondria.


Assuntos
Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/toxicidade , Infecções por Helicobacter/metabolismo , Helicobacter pylori/metabolismo , Alvo Mecanístico do Complexo 1 de Rapamicina/efeitos dos fármacos , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Aminoácidos , Animais , Autofagia/efeitos dos fármacos , Proteína Homóloga à Proteína-1 Relacionada à Autofagia/metabolismo , Toxinas Bacterianas/metabolismo , Linhagem Celular , Feminino , Células HEK293 , Homeostase , Interações Hospedeiro-Patógeno/fisiologia , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo
16.
Reprod Biol ; 18(1): 115-121, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29449095

RESUMO

The Aim of this study was to evaluate the effects of bacteriospermia on human sperm parameters, nuclear protamines, DNA integrity and ICSI outcome in patients enrolled for ICSI treatment. 84 unselected couples consulting in infertility and obstetrics clinic and enrolled for ICSI treatment were included in this study. The semen specimens were screened bacteriologically; semen and sperm parameters were also evaluated according to WHO guidelines. DNA integrity, protamines concentration and protamine deficiency were estimated by TUNEL assay, AU-PAGE and Chromomycin (CMA3) respectively. The results of this study revealed that 34.52% of studied semen samples were infected with bacteria. The isolated bacteria were identified as Staphylococcus aureus, Staph. epidermidis, Staph. haemolyticus, Escherichia coli, Enterococcus faecalis and Streptococcus agalactiae. Bacteriospermia had a significant (p < .010) negative effect on sperm parameters; concentration, motility, progressive motility and chromatin condensation. Moreover, high DNA fragmentation with low P1 and P2 concentrations were noticed in infected patients in comparison to non-infected patients but non-significant. Also, the fertilization rate decreased significantly (p < .05) with infected patients. IN CONCLUSION: bacteriospermia has significant negative effect on sperm quality and fertilization rate in patients who underwent ICSI treatment.


Assuntos
Núcleo Celular/metabolismo , Fragmentação do DNA , Infertilidade Masculina/etiologia , Protaminas/metabolismo , Infecções do Sistema Genital/fisiopatologia , Injeções de Esperma Intracitoplásmicas , Espermatozoides/metabolismo , Adulto , Núcleo Celular/microbiologia , Núcleo Celular/patologia , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/patologia , Infecções por Escherichia coli/fisiopatologia , Características da Família , Feminino , Alemanha/epidemiologia , Infecções por Bactérias Gram-Positivas/epidemiologia , Infecções por Bactérias Gram-Positivas/microbiologia , Infecções por Bactérias Gram-Positivas/patologia , Infecções por Bactérias Gram-Positivas/fisiopatologia , Hospitais Universitários , Humanos , Infertilidade Feminina/terapia , Masculino , Pessoa de Meia-Idade , Gravidez , Taxa de Gravidez , Infecções do Sistema Genital/epidemiologia , Infecções do Sistema Genital/microbiologia , Infecções do Sistema Genital/patologia , Sêmen/microbiologia , Análise do Sêmen , Espermatozoides/microbiologia , Espermatozoides/patologia
17.
J Anal Methods Chem ; 2017: 8489471, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28894622

RESUMO

Enalapril is an angiotensin-converting enzyme inhibitor used for treatment of hypertension and chronic heart disease. Enalaprilat is its active metabolite responsible for the activity. This study aimed to develop and validate a method for enalapril and enalaprilat analysis and to determine the bioequivalence of two tablet formulae of enalapril. LC-MS/MS bioanalytical method was developed and validated and then applied to evaluate the bioavailability of two enalapril formulae. Antihyperglycemic sitagliptin was used as internal standard (IS). The method was accurate for the within- and between-days analysis, and precise CV% was <5%, being linear over the calibration range 1.0-200.0 ng/ml. Stability was >85% and the LOD was 0.907 and 0.910 ng/ml for enalapril and enalaprilat, respectively, and LLOQ was 1 ng/ml. The pharmacokinetic parameters Cmax, tmax, AUC0-72, and AUC0-∞ values of enalapril and enalaprilat of the two formulae were calculated and nonsignificant differences were found. A linearity, specific, accurate, and precise method was developed and applied for the analysis of enalapril and enalaprilat in human plasma after oral administration of two formulae of enalapril 20 mg tablets in healthy volunteers. Depending on the statistical analysis it was concluded that the two enalapril formulae were bioequivalent.

18.
Intervirology ; 60(4): 156-164, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29428951

RESUMO

OBJECTIVE: This study aims to provide a molecular and epidemiological characterization of bovine papillomavirus (BPV) infections in Iraq. METHODS: The present study focuses on identifying BPV based on clinical and epidemiological manifestations, histopathological examinations, and polymerase chain reactions (PCR). Samples were collected from 163 animals suffering from cutaneous bovine papillomatosis, including 129 females (79.14%) with an age range of 16-40 months and 34 males (20.85%) with an age range of 17-29 months. RESULTS: The incidence rate was significantly higher in females than in males. The most commonly affected sites were the teats and neck, though warts were found in other areas of the body. Histological sections were diagnosed as fibropapilloma. PCR results showed that 80.13% of the extracted papilloma DNA samples corresponded to the BPV-1 genotype. Furthermore, 7.94% of the samples showed a mixed infection of BPV-1 and BPV-13. While, 40.63% of the extracted DNA blood samples showed 2 DNA fragments corresponding to both genotypes BPV-1 and BPV-2. CONCLUSIONS: This study confirmed the presence of BPV-1, BPV-2, and BPV-13, which belong to the Deltapapillomavirus genera, for the first time in the DNA of Iraqi cattle. Understanding BPV diversity and epidemiology is of critical importance for starting prevention strategies.


Assuntos
Doenças dos Bovinos/epidemiologia , Genótipo , Papiloma/veterinária , Papillomaviridae/classificação , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/veterinária , Animais , Bovinos , Doenças dos Bovinos/patologia , Doenças dos Bovinos/virologia , Feminino , Histocitoquímica , Iraque/epidemiologia , Masculino , Epidemiologia Molecular , Papiloma/epidemiologia , Papiloma/patologia , Papiloma/virologia , Infecções por Papillomavirus/epidemiologia , Infecções por Papillomavirus/patologia , Infecções por Papillomavirus/virologia , Reação em Cadeia da Polimerase
19.
Adv Virol ; 2016: 2143024, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27413374

RESUMO

This study aimed to provide the first molecular characterization of bovine papillomavirus type 1 (BPV-1) in Iraq. BPV is a widely spread oncogenic virus in Iraqi cattle and is associated with the formation of both benign and malignant lesions, resulting in notable economic losses in dairy and beef cattle. In the current study, 140 cutaneous papilloma specimens were collected from cattle in central Iraq. These samples were submitted to histopathological examination, PCR, and sequencing analysis. The histopathology revealed that the main lesion type among the specimens was fibropapilloma. BPV-1 DNA was detected in 121 of the samples (86.42%) in Iraqi cattle as the main causative agent for the disease. A partial sequence for the E2, L2 genes, and complete sequence for the E5 gene were deposited in GenBank. Phylogenetic analysis confirmed the presence of BPV-1 and showed that the origin of infection may be imported European cattle. Obtaining a complete E5 gene sequence enabled us to perform structural predictions. This study presents the first report of BPV-1 infection in the Iraqi cattle and contributes to extending the knowledge of the origin of the spread of this disease. The results of this study will aid in the development of appropriate control measures and therapeutic strategies.

20.
Biochem Res Int ; 2016: 8920503, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27110402

RESUMO

Vitamin D is necessary for maintaining and regulating calcium levels; thus, insufficiency of vitamin D increases the risk of many chronic diseases. This study aimed to examine vitamin D levels among Jordanian and Iraqi volunteers and find the relation between vitamin D level and lipid profile patients. Vitamin D levels were evaluated using enzyme-linked immunosorbent assay. For young healthy group subjects, vitamin D levels were 20.60 ± 5.94 ng/mL for Jordanian and 27.59 ± 7.74 ng/mL for Iraqi. Vitamin D concentrations for young males and females were 25.82 ± 8.33 ng/mL and 21.95 ± 6.39 ng/mL, respectively. Females wearing hijab were 20.87 ± 6.45 ng/mL, while uncovered females were 23.55 ± 6.04 ng/mL. For >40 years Iraqi subjects, vitamin D level for healthy was 29.78 ± 9.49 ng/mL and 23.88 ± 7.93 ng/mL for hyperlipidemic subjects. Vitamin D levels for overweight and obese healthy groups were significantly higher (P < 0.050) than those for the hyperlipidemic patients groups. Vitamin D levels for males were significantly higher than females and were significantly higher for healthy than those hyperlipidemic Iraqi patients. These findings showed that vitamin D levels are affected by age, nationality, gender, and health statues and highlight the importance of vitamin D supplementation for groups with low levels particularly old, hijab wearing females, and hyperlipidemic groups.

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