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1.
Int J Mycobacteriol ; 13(2): 221-224, 2024 Apr 01.
Article in English | MEDLINE | ID: mdl-38916396

ABSTRACT

Nocardiosis is an opportunistic infection that affects both immunocompromised as well as immunocompetent patients. The main infections occur as soft tissue and lung infections although they might disseminate to various organs. This is a case study aimed to reflect the severity of the disease and the patient's risk factors associated with the infection. A sputum sample was collected from tuberculosis (TB) suspects for culture. Nocardia-like colonies were isolated, purified, and sent to BGI Company (Hongkong, China). Standard forward sequencing of 16S rRNA was done by ABI Genetic Analyzer (Applied Biosystems). Sequence alignment and nucleotide basic local alignment search tool (BLAST) were done using National Center for Bioinformatics (NCBI) Nucleotide BLAST. Biochemical identification to the colonies was done using an automation system (BD Phoenix™) to confirm the identification. Nocardia paucivorans was identified from the TB suspect. Risk factors were identified as extensive contact to dust, absence of primary care units with complete facilities, and old age. Since the infection of the lungs caused by Nocardia might be similar to pulmonary TB, this case report highlights the importance of accurate diagnosis and identification procedures to differentiate between the two.


Subject(s)
Nocardia Infections , Nocardia , RNA, Ribosomal, 16S , Sputum , Humans , Nocardia Infections/microbiology , Nocardia Infections/diagnosis , Nocardia/isolation & purification , Nocardia/genetics , Male , Fatal Outcome , Sputum/microbiology , RNA, Ribosomal, 16S/genetics , Middle Aged , Respiratory Tract Infections/microbiology , Respiratory Tract Infections/diagnosis , Gold , Risk Factors , Tuberculosis, Pulmonary/microbiology , Tuberculosis, Pulmonary/diagnosis
2.
Drug Deliv Transl Res ; 13(11): 2847-2868, 2023 11.
Article in English | MEDLINE | ID: mdl-37184748

ABSTRACT

Oral candidiasis (OC) is an opportunistic fungal infection, common amongst the elderly and the immunocompromised. Unfortunately, the therapeutic efficacy of common antifungals is imperiled by the rise of antifungal drug resistance. An alternative promising therapeutic option possibly contributing to antifungal therapy is drug repurposing. Herein, we aimed to employ novel pharmaceutical drug delivery for enhancing the emerging antifungal potential of the hypocholesterolemic drug atorvastatin (ATV). ATV-propylene-glycol-liposomes (ATV/PG-Lip) were prepared then integrated in 3D-printed (3DP) mucoadhesive films comprising chitosan, polyvinyl-alcohol and hydroxypropyl methylcellulose, as an innovative blend, for the management of OC. ATV/PG-Lip demonstrated good colloidal properties of particle size (223.3 ± 2.1 nm), PDI (0.12 ± 0.001) and zeta potential (-18.2 ± 0.3 mV) with high entrapment efficiency (81.15 ± 1.88%) and sustained drug release. Also, ATV/PG-Lip showed acceptable three-month colloidal stability and in vitro cytocompatibility on human gingival fibroblasts. The developed 3DP-films exhibited controlled ATV release (79.4 ± 1.4% over 24 h), reasonable swelling and mucoadhesion (2388.4 ± 18.4 dyne/cm2). In vitro antifungal activity of ATV/PG-Lip was confirmed against fluconazole-resistant Candida albicans via minimum inhibitory concentration determination, time-dependent antifungal activity, agar diffusion and scanning electron microscopy. Further, ATV/PG-Lip@3DP-film exceeded ATV@3DP-film in amelioration of infection and associated inflammation in an in vivo oral candidiasis rabbit model. Accordingly, the results confirm the superiority of the fabricated ATV/PG-Lip@3DP-film for the management of oral candidiasis and tackling antifungal resistance.


Subject(s)
Candidiasis, Oral , Animals , Humans , Rabbits , Aged , Candidiasis, Oral/drug therapy , Antifungal Agents , Liposomes/therapeutic use , Atorvastatin , Polymers/therapeutic use , Drug Repositioning , Printing, Three-Dimensional
3.
J Clin Virol ; 162: 105422, 2023 05.
Article in English | MEDLINE | ID: mdl-36989731

ABSTRACT

BACKGROUND: The COVID-19 pandemic led to severe health systems collapse, as well as logistics and supply delivery shortages across sectors. Delivery of PCR related healthcare supplies continue to be hindered. There is the need for a rapid and accessible SARS-CoV-2 molecular detection method in low resource settings. OBJECTIVES: To validate a novel isothermal amplification method for rapid detection of SARS-CoV-2 across seven sub-Sharan African countries. STUDY DESIGN: In this multi-country phase 2 diagnostic study, 3,231 clinical samples in seven African sites were tested with two reverse transcription Recombinase-Aided Amplification (RT-RAA) assays (based on SARS-CoV-2 Nucleocapsid (N) gene and RNA-dependent RNA polymerase (RdRP) gene). The test was performed in a mobile suitcase laboratory within 15 min. All results were compared to a real-time RT-PCR assay. Extraction kits based on silica gel or magnetic beads were applied. RESULTS: Four sites demonstrated good to excellent agreement, while three sites showed fair to moderate results. The RdRP gene assay exhibited an overall PPV of 0.92 and a NPV of 0.88. The N gene assay exhibited an overall PPV of 0.93 and a NPV 0.88. The sensitivity of both RT-RAA assays varied depending on the sample Ct values. When comparing sensitivity between sites, values differed considerably. For high viral load samples, the RT-RAA assay sensitivity ranges were between 60.5 and 100% (RdRP assay) and 25 and 98.6 (N assay). CONCLUSION: Overall, the RdRP based RT-RAA test showed the best assay accuracy. This study highlights the challenges of implementing rapid molecular assays in field conditions. Factors that are important for successful deployment across countries include the implementation of standardized operation procedures, in-person continuous training for staff, and enhanced quality control measures.


Subject(s)
COVID-19 , SARS-CoV-2 , Humans , SARS-CoV-2/genetics , COVID-19/diagnosis , Pandemics , Sensitivity and Specificity , Nucleic Acid Amplification Techniques/methods , Real-Time Polymerase Chain Reaction , Africa South of the Sahara , RNA, Viral/genetics
4.
Biology (Basel) ; 12(3)2023 Feb 21.
Article in English | MEDLINE | ID: mdl-36979037

ABSTRACT

Streptomyces are factories of antimicrobial secondary metabolites. We isolated a Streptomyces species associated with the Pelargonium graveolens rhizosphere. Its total metabolic extract exhibited potent antibacterial and antifungal properties against all the tested pathogenic microbes. Whole genome sequencing and genome analyses were performed to take a look at its main characteristics and to reconstruct the metabolic pathways that can be associated with biotechnologically useful traits. AntiSMASH was used to identify the secondary metabolite gene clusters. In addition, we searched for known genes associated with plant growth-promoting characteristics. Finally, a comparative and pan-genome analysis with three closely related genomes was conducted. It was identified as Streptomyces vinaceusdrappus strain AC-40. Genome mining indicated the presence of several secondary metabolite gene clusters. Some of them are identical or homologs to gene clusters of known metabolites with antimicrobial, antioxidant, and other bioactivities. It also showed the presence of several genes related to plant growth promotion traits. The comparative genome analysis indicated that at least five of these gene clusters are highly conserved through rochei group genomes. The genotypic and phenotypic characteristics of S. vinaceusdrappus strain AC-40 indicate that it is a promising source of beneficial secondary metabolites with pharmaceutical and biotechnological applications.

5.
Front Microbiol ; 11: 590776, 2020.
Article in English | MEDLINE | ID: mdl-33329469

ABSTRACT

IncP-1 plasmids, first isolated from clinical specimens (R751, RP4), are recognized as important vectors spreading antibiotic resistance genes. The abundance of IncP-1 plasmids in the environment, previously reported, suggested a correlation with anthropogenic pollution. Unexpectedly, qPCR-based detection of IncP-1 plasmids revealed also an increased relative abundance of IncP-1 plasmids in total community DNA from the rhizosphere of lettuce and tomato plants grown in non-polluted soil along with plant age. Here we report the successful isolation of IncP-1 plasmids by exploiting their ability to mobilize plasmid pSM1890. IncP-1 plasmids were captured from the rhizosphere but not from bulk soil, and a high diversity was revealed by sequencing 14 different plasmids that were assigned to IncP-1ß, δ, and ε subgroups. Although backbone genes were highly conserved and mobile elements or remnants as Tn501, IS1071, Tn402, or class 1 integron were carried by 13 of the sequenced IncP-1 plasmids, no antibiotic resistance genes were found. Instead, seven plasmids had a mer operon with Tn501-like transposon and five plasmids contained putative metabolic gene clusters linked to these mobile elements. In-depth sequence comparisons with previously known plasmids indicate that the IncP-1 plasmids captured from the rhizosphere are archetypes of those found in clinical isolates. Our findings that IncP-1 plasmids do not always carry accessory genes in unpolluted rhizospheres are important to understand the ecology and role of the IncP-1 plasmids in the natural environment.

6.
Front Microbiol ; 11: 149, 2020.
Article in English | MEDLINE | ID: mdl-32132980

ABSTRACT

PromA plasmids are broad host range (BHR) plasmids, which are often cryptic and hence have an uncertain ecological role. We present three novel PromA γ plasmids which carry genes associated with degradation of the phenylurea herbicide linuron, two of which originated from unrelated Hydrogenophaga hosts isolated from different environments (pPBL-H3-2 and pBPS33-2), and one (pEN1) which was exogenously captured from an on-farm biopurification system (BPS). Hydrogenophaga sp. plasmid pBPS33-2 carries all three necessary gene clusters (hylA, dca, ccd) determining the three main steps for conversion of linuron to Krebs cycle intermediates, while pEN1 only determines the initial linuron hydrolysis step. Hydrogenophaga sp. plasmid pPBL-H3-2 exists as two variants, both containing ccd but with the hylA and dca gene modules interchanged between each other at exactly the same location. Linuron catabolic gene clusters that determine the same step were identical on all plasmids, encompassed in differently arranged constellations and characterized by the presence of multiple IS1071 elements. In all plasmids except pEN1, the insertion spot of the catabolic genes in the PromA γ plasmids was the same. Highly similar PromA plasmids carrying the linuron degrading gene cargo at the same insertion spot were previously identified in linuron degrading Variovorax sp. Interestingly, in both Hydrogenophaga populations not every PromA plasmid copy carries catabolic genes. The results indicate that PromA plasmids are important vehicles of linuron catabolic gene dissemination, rather than being cryptic and only important for the mobilization of other plasmids.

7.
Front Microbiol ; 10: 2835, 2019.
Article in English | MEDLINE | ID: mdl-31998244

ABSTRACT

Ralstonia solanacearum (biovar2, race3) is the causal agent of bacterial wilt and this quarantine phytopathogen is responsible for massive losses in several commercially important crops. Biological control of this pathogen might become a suitable plant protection measure in areas where R. solanacearum is endemic. Two bacterial strains, Bacillus velezensis (B63) and Pseudomonas fluorescens (P142) with in vitro antagonistic activity toward R. solanacearum (B3B) were tested for rhizosphere competence, efficient biological control of wilt symptoms on greenhouse-grown tomato, and effects on the indigenous rhizosphere prokaryotic communities. The population densities of B3B and the antagonists were estimated in rhizosphere community DNA by selective plating, real-time quantitative PCR, and R. solanacearum-specific fliC PCR-Southern blot hybridization. Moreover, we investigated how the pathogen and/or the antagonists altered the composition of the tomato rhizosphere prokaryotic community by 16S rRNA gene amplicon sequencing. B. velezensis (B63) and P. fluorescens (P142)-inoculated plants showed drastically reduced wilt disease symptoms, accompanied by significantly lower abundance of the B3B population compared to the non-inoculated pathogen control. Pronounced shifts in prokaryotic community compositions were observed in response to the inoculation of B63 or P142 in the presence or absence of the pathogen B3B and numerous dynamic taxa were identified. Confocal laser scanning microscopy (CLSM) visualization of the gfp-tagged antagonist P142 revealed heterogeneous colonization patterns and P142 was detected in lateral roots, root hairs, epidermal cells, and within xylem vessels. Although competitive niche exclusion cannot be excluded, it is more likely that the inoculation of P142 or B63 and the corresponding microbiome shifts primed the plant defense against the pathogen B3B. Both inoculants are promising biological agents for efficient control of R. solanacearum under field conditions.

8.
Int J Mycobacteriol ; 7(3): 236-241, 2018.
Article in English | MEDLINE | ID: mdl-30198502

ABSTRACT

Background: The aim of this study was to characterize the drug resistance profile, and the specific lineages of Mycobacterium tuberculosis (MTB) strains isolated from patients with pulmonary TB in the state of Khartoum in Sudan. Methods: Consecutive sputum samples and clinical data were collected from 406 smear-positive TB patients with pulmonary TB in 2007-2009. The samples were cultured, and drug susceptibility testing (DST) was performed using the proportion method (PM) on solid Löwenstein-Jensen medium, and species were identified using biochemical methods at the National Reference Laboratory (NRL) in Khartoum. Extracted deoxyribonucleic acid from a total of 120, 60 suspected multidrug-resistant isolates (MDR), and 60 non-MDR isolates were subsequently sent to the WHO supranational reference laboratory (SRL) in Stockholm at the Public Health Agency of Sweden, for confirmation of the drug resistance profile, examinations by line probe assay (LPA), and molecular epidemiology analysis with Spoligotyping. Results: LPA results correlated 100% for non-MDR and 62% for the suspected MDR strains when compared to the DST results obtained by PM at the NRL. Two strains were initially using the PM identified as MDR-TB but later shown by Hain GenoType Mycobacterium CM/AS to belong Mycobacterium avium complex (Mycobacterium intracelluare). These two strains were excluded from the study material for further analysis. The remaining 58 MDR strains were analyzed using LPA, and 36 strains were confirmed as MDR, 10 as rifampicin monoresistant, and eight as isoniazid-monoresistant. Spoligotyping for all the 118 MTB isolates revealed a total of 115 patterns in which four patterns represented major clusters with a total of 108 (91%) of the strains. The CAS1_Delhi/family was the predominant type and detected in 62 isolates (52%), of which 26 were MDR and 36 were susceptible. It was followed by H3/family with 19 (16%) strains, and 11 Latin American Mediterranean3/family, 16 T2/T1, and two strains each of the Beijing and S lineage. Conclusion: Comparison of DST results obtained using PM and LPA showed 100% agreement for the non-MDR strains but only 62% for the MDR strains. Taking in consideration the time, risk of contamination and the cost of labour to identify MDR TB, the LPA have clear advantages in early detection of MDRTB than the PM. Additionally in this study material Spoligotyping revealed the CAS1 Delhi as the most predominant family. We could not see no major difference in lineages between MDR and non-MDR strains.


Subject(s)
Mycobacterium tuberculosis/genetics , Tuberculosis, Pulmonary/epidemiology , Tuberculosis, Pulmonary/microbiology , Antitubercular Agents/pharmacology , Bacterial Typing Techniques , DNA, Bacterial/genetics , Genotype , Humans , Isoniazid/pharmacology , Microbial Sensitivity Tests , Mycobacterium tuberculosis/drug effects , Mycobacterium tuberculosis/isolation & purification , Rifampin/pharmacology , Sputum , Sudan/epidemiology , Tuberculosis, Multidrug-Resistant/epidemiology , Tuberculosis, Multidrug-Resistant/microbiology
9.
Pain Physician ; 21(2): 191-198, 2018 03.
Article in English | MEDLINE | ID: mdl-29565949

ABSTRACT

BACKGROUND: Pulsed radiofrequency (PRF) is increasingly used in clinical practice, especially in neuropathic pain disorders. Although PRF is not new to clinical use, there are significant gaps in knowledge regarding its effectiveness. OBJECTIVES: The current study was conducted to evaluate the effect of duration of application of PRF on its analgesic efficacy in improvement of neuropathic pain. STUDY DESIGN: A randomized experimental trial. SETTING: An animal research facility at the College of Veterinary Medicine at Mansoura University in Egypt. METHODS: Chronic constriction of the sciatic nerve of 36 male Sprague-Dawley rats was performed to induce neuropathic pain. The rats were divided into 6 groups (6 rats each) in which PRF was applied for 2, 4, 6, or 8 minutes or not at all. In one group, RF cannula was applied without performing PRF intervention. The pain was assessed through observation of resting paw posture (RPP) at 3, 10, and 21 days. Nerve damage was assessed by histopathological evaluation of the sciatic nerve. Immunohistochemical localization of proinflammatory cytokines (interleukin 6 [IL-6] and tumor necrosis factor alpha [TNF-alpha]) was also done. RESULTS: RPP was improved in rats treated with PRF. This improvement was significant only in rats treated for 8 minutes. Increased duration for PRF application was associated with a significant decrease in IL-6 and TNF-alpha contents in all groups when compared with the control group. Histopathological evaluation of the constricted sciatic nerve revealed no statistical significance among the different study groups. LIMITATIONS: The study was limited by the lack of measurement of other inflammatory markers that may help elucidate other relevant mechanisms. CONCLUSIONS: Increased duration of PRF application resulted in better analgesic efficacy without any increase in tissue injury in an animal model of neuropathic pain. This effect may be attributed to decreased production of pro-inflammatory cytokines. KEY WORDS: Pulsed radiofrequency, analgesic, rats, sciatic nerve, duration, neuropathic pain.


Subject(s)
Neuralgia , Pulsed Radiofrequency Treatment/methods , Animals , Disease Models, Animal , Male , Rats , Rats, Sprague-Dawley , Sciatic Nerve/injuries
10.
Appl Microbiol Biotechnol ; 101(11): 4815-4825, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28235988

ABSTRACT

On-farm biopurification systems (BPSs) represent an efficient technology for treating pesticide-contaminated wastewater. Biodegradation by genetically adapted bacteria has been suggested to perform a major contribution to the removal of pesticides in BPSs. Recently, several studies pointed to the role of IncP-1 plasmids in the degradation of pesticides in BPSs but this was never linked with catabolic markers. Therefore, a microcosm experiment was conducted in order to examine whether changes in mobile genetic element (MGE) abundances in response to the application of phenylurea herbicide linuron are linked with changes in catabolic genes. Denaturing gradient gel electrophoresis (DGGE) fingerprints of 16S ribosomal RNA gene fragments amplified from total community (TC)-DNA suggested significant shifts in the bacterial community composition. PCR-Southern blot-based detection of genes involved in linuron hydrolysis (libA and hylA) or degradation of its metabolite 3,4-dichloroaniline (dcaQ I , dcaQ II , and ccdC) in TC-DNA showed that the abundance of the hylA gene was increased faster and stronger in response to linuron application than that of the libA gene, and that the dcaQ II gene was more abundant than the isofunctional gene dcaQ I 20 and 60 days after linuron addition. Furthermore, a significant increase in the relative abundance of the IncP-1-specific korB gene in response to linuron was recorded. Our data suggest that different bacterial populations bearing isofunctional genes coding for enzymes degrading linuron seemed to be enriched in BPSs in response to linuron and that IncP-1 plasmids might be involved in their dissemination.


Subject(s)
Linuron/metabolism , Microbial Consortia/genetics , Pesticides/metabolism , Soil Microbiology , Agriculture , Biodegradation, Environmental , Comamonadaceae/drug effects , Comamonadaceae/genetics , DNA, Bacterial , Denaturing Gradient Gel Electrophoresis , Hydrolysis , Interspersed Repetitive Sequences , Linuron/pharmacology , Microbial Consortia/drug effects , Plasmids , Polymerase Chain Reaction , RNA, Ribosomal, 16S , Wastewater
11.
FEMS Microbiol Ecol ; 92(2)2016 Feb.
Article in English | MEDLINE | ID: mdl-26705572

ABSTRACT

On-farm biopurification systems (BPSs) treat pesticide-contaminated wastewater at farms through biodegradation and sorption processes. However, information on the microbiota involved in pesticide removal in BPSs is scarce. Here we report on the response of BPS bacterial communities to the herbicide linuron (BPS(+)) compared with the control (BPS(-)) in a microcosm experiment. Both denaturing gradient gel electrophoresis (DGGE) and pyrosequencing of 16S rRNA gene fragments amplified from community DNA indicated shifts in the bacterial community after linuron application. Responding populations belonged to taxa that were previously reported from linuron degrading consortia cultivated from soil (Hyphomicrobiaceae, Comamonadaceae, Micrococcaceae). In addition, numerous taxa with increased relative abundance were identified that were previously not associated with linuron degradation. The relative abundance of IncP-1 korB copies increased in response to linuron application. Amplicon pyrosequencing of IncP-1 trfA genes revealed a high IncP-1 plasmid diversity and suggested that populations carrying IncP-1ß plasmids increased in relative abundance. Transferable mercury resistance plasmids were exogenously captured from BPS(+)/BPS(-), and in three transconjugants from BPS(+) the gene hylA was detected. Our data suggest the existence of a multispecies linuron degrading bacterial food web and an involvement of IncP-1 plasmids in the adaptation of bacterial communities to pesticide pollution in BPSs.


Subject(s)
Comamonadaceae/genetics , Comamonadaceae/metabolism , Herbicides/metabolism , Herbicides/pharmacology , Linuron/metabolism , Linuron/pharmacology , Biodegradation, Environmental , Comamonadaceae/drug effects , DNA, Bacterial/genetics , Denaturing Gradient Gel Electrophoresis , Molecular Sequence Data , Plasmids/genetics , RNA, Ribosomal, 16S/genetics , Soil , Soil Microbiology , Wastewater/microbiology
12.
Plasmid ; 80: 16-23, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25957823

ABSTRACT

Biopurification systems (BPS) are complex soil-related and artificially-generated environments usually designed for the removal of toxic compounds from contaminated wastewaters. The present study has been conducted to isolate and characterize a collection of cultivable plasmid-carrying bacterial isolates recovered from a BPS established for the decontamination of wastewater generated in a farmyard. Out of 1400 isolates, a collection of 75 plasmid-containing bacteria was obtained, of which 35 representative isolates comprising in total at least 50 plasmids were chosen for further characterization. Bacterial hosts were taxonomically assigned by 16S ribosomal RNA gene sequencing and phenotypically characterized according to their ability to grow in presence of different antibiotics and heavy metals. The study demonstrated that a high proportion of the isolates was tolerant to antibiotics and/or heavy metals, highlighting the on-farm BPS enrichment in such genetic traits. Several plasmids conferring such resistances in the bacterial collection were detected to be either mobilizable or selftransmissible. Occurrence of broad host range plasmids of the incompatibility groups IncP, IncQ, IncN and IncW was examined with positive results only for the first group. Presence of the IS1071 insertion sequence, frequently associated with xenobiotics degradation genes, was detected in DNA obtained from 24 of these isolates, strongly suggesting the presence of yet-hidden catabolic activities in the collection of isolates. The results showed a remarkable diversity in the plasmid mobilome of cultivable bacteria in the BPS with the presence of abundant resistance markers of different types, thus providing a suitable environment to investigate the genetic structure of the mobile genetic pool in a model on-farm biofilter for wastewater decontamination in intensive agricultural production.


Subject(s)
Gammaproteobacteria/isolation & purification , Plasmids/genetics , Agriculture , Biodegradation, Environmental , DNA, Bacterial/genetics , Gammaproteobacteria/drug effects , Gammaproteobacteria/genetics , Metals, Heavy/pharmacology , Microbial Sensitivity Tests , Molecular Typing , Pesticide Residues/isolation & purification , RNA, Ribosomal, 16S/genetics , Soil Microbiology , Water Pollutants, Chemical/isolation & purification , Water Purification
13.
Am J Microbiol Res ; 3(4): 125-128, 2015.
Article in English | MEDLINE | ID: mdl-30294615

ABSTRACT

Tuberculosis (TB) remains a major public health problem worldwide due to its high risk of person-to-person transmission, morbidity and mortality [1]. Sudan has a high burden of tuberculosis. Spoligotyping (spacer oligonucleotide typing) a rapid method for genotyping of Mycobacterium tuberculosis using the principle of reverse hybridization. The ecology of the prevalent mycobacteria strain can vary depending on country and region. The aim of this study was to determine the genotyping of Mycobacterium tuberculosis isolated from Sudan using spoligotyping SPOLDB4. A total of 75 Mycobacterium tuberculosis sputum samples were collected from pulmonary Tuberculosis patients attending references Laboratories and diagnostic centers in Khartoum and Eastern Sudan in (2011-2013). The mycobacteria were genotyped using Spoligotyping technique and data obtained were analyzed and compared to the SPOLDB4 database. Among the 75 isolate analyzed, 57(76%) were identified by SPOLDB4 and 18 (24%) could not be matched to any lineages. The most prevalent genotype cluster was MANU2 38 (50.7%) followed by CASI Delhi 8 (10.7%). In the study SIT54 was the most common pattern 37 (49.3%) followed by SIT25 6(8%).

14.
Arch Med Res ; 43(7): 555-62, 2012 Oct.
Article in English | MEDLINE | ID: mdl-23085447

ABSTRACT

BACKGROUND AND AIMS: Hepatitis C virus (HCV) has emerged as the major pathogen of liver disease worldwide. The aim of this study was to quantitate and evaluate the performance of HCV-NS4 antigen as an alternative approach for confirmation of viremia. METHODS: Detection of HCV-NS4 was assessed in 883 patients with chronic hepatitis C. Areas under the ROC curves (AUC) were used to assess and compare diagnostic accuracy of ELISA for HCV-NS4 with quantitative HCV-RNA as a gold standard. RESULTS: HCV-NS4 was identified at 27 kDa using Western blot. AUC for HCV-NS4 detection was 0.95 for all patients with different liver pathologies: 0.93 for liver fibrosis (LF), 0.95 for liver cirrhosis (LC) and 0.98 for hepatocellular carcinoma (HCC). The mean ± SD (µg/mL) of HCV-NS4 in LF was 94.2 ± 55.6; in LC was 99.3 ± 64.8 and in HCC was 124.9 ± 70.3. CONCLUSIONS: HCV-NS4 antigen detection using ELISA is a reliable test in the confirmation of HCV infection.


Subject(s)
Hepatitis C Antigens/analysis , Hepatitis C, Chronic/diagnosis , Liver Diseases/virology , Viral Nonstructural Proteins/analysis , Adult , Blotting, Western , Carcinoma, Hepatocellular/complications , Carcinoma, Hepatocellular/immunology , Carcinoma, Hepatocellular/virology , Egypt , Enzyme-Linked Immunosorbent Assay , Female , Hepacivirus/immunology , Hepacivirus/isolation & purification , Hepatitis C Antigens/immunology , Hepatitis C, Chronic/complications , Hepatitis C, Chronic/immunology , Humans , Liver Cirrhosis/complications , Liver Cirrhosis/immunology , Liver Cirrhosis/virology , Liver Diseases/complications , Liver Diseases/immunology , Liver Neoplasms/complications , Liver Neoplasms/immunology , Liver Neoplasms/virology , Male , Middle Aged , Viral Nonstructural Proteins/immunology , Viremia/complications , Viremia/diagnosis , Viremia/immunology , Young Adult
15.
Arab J Urol ; 10(2): 199-206, 2012 Jun.
Article in English | MEDLINE | ID: mdl-26558026

ABSTRACT

OBJECTIVES: To evaluate and compare the relative contribution of different therapeutic agents for renoprotection against complete unilateral ureteric obstruction (UUO), using a rabbit model sampled at different times. MATERIALS AND METHODS: Eighty-four male New Zealand White rabbits were divided into seven groups of 12 rabbits; a sham group, a control (left UUO + no medication) or left UUO and treated with either enalapril, losartan, verapamil, l-arginine or antioxidant (vitamin E and selenium mixture). Rabbits in the control and treated groups were subjected to 3, 10 and 21 days of complete ureteric ligation and then killed humanely. The control and treated groups were evaluated at baseline and at the end of the experiment, by measuring split effective renal plasma flow (ERPF) using diuretic renography, and the split glomerular filtration rate (GFR) using selective creatinine clearance. Renal histopathology was evaluated using a tubulo-interstitial damage score. RESULTS: In the sham group there was no significant effect on any of the evaluated variables. For split ERPF, losartan showed the highest renoprotective effect, saving 44% and 77% of ERPF at 3 and 21 days after UUO, respectively. Losartan was also the best renoprotective agent for GFR. For renal histopathology, enalapril showed the earliest and greatest improvement as assessed by the damage score, reaching 60% at 21 days after UUO. l-Arginine was the next best effect to blockade the renin-angiotensin system for renoprotection. CONCLUSION: We suggest that blockade of the renin-angiotensin system provides the best renoprotection against the effects of complete UUO.

16.
Ther Apher Dial ; 13(3): 220-4, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19527469

ABSTRACT

Metabolic acidosis can occur as a result of either the accumulation of endogenous acids or loss of bicarbonate from the gastrointestinal tract or the kidney, which represent common causes of metabolic acidosis. The appropriate treatment of acute metabolic acidosis has been very controversial. Ionized alkaline water was not evaluated in such groups of patients in spite of its safety and reported benefits. So, we aimed to assess its efficacy in the management of metabolic acidosis in animal models. Two models of metabolic acidosis were created in dogs and rats. The first model of renal failure was induced by ligation of both ureters; and the second model was induced by urinary diversion to gut (gastrointestinal bicarbonate loss model). Both models were subjected to ionized alkaline water (orally and by hemodialysis). Dogs with renal failure were assigned to two groups according to the type of dialysate utilized during hemodialysis sessions, the first was utilizing alkaline water and the second was utilizing conventional water. Another two groups of animals with urinary diversion were arranged to receive oral alkaline water and tap water. In renal failure animal models, acid-base parameters improved significantly after hemodialysis with ionized alkaline water compared with the conventional water treated with reverse osmosis (RO). Similar results were observed in urinary diversion models as there was significant improvement of both the partial pressure of carbon dioxide and serum bicarbonate (P = 0.007 and 0.001 respectively) after utilizing alkaline water orally. Alkaline ionized water can be considered as a major safe strategy in the management of metabolic acidosis secondary to renal failure or dialysis or urinary diversion. Human studies are indicated in the near future to confirm this issue in humans.


Subject(s)
Acidosis/therapy , Renal Dialysis , Renal Insufficiency/complications , Water/administration & dosage , Acidosis/etiology , Administration, Oral , Animals , Bicarbonates/blood , Bicarbonates/metabolism , Carbon Dioxide/metabolism , Disease Models, Animal , Dogs , Hydrogen-Ion Concentration , Partial Pressure , Rats , Water/adverse effects , Water/chemistry
17.
BJU Int ; 99(5): 1098-102, 2007 May.
Article in English | MEDLINE | ID: mdl-17233810

ABSTRACT

OBJECTIVE: To evaluate the efficacy of oxidized cellulose, Surgicel(TM) (Johnson & Johnson Medical, New Brunswick, NJ, USA) for patching defects in the tunica albuginea in a rabbit model, with a future application to correct chordee. MATERIALS AND METHODS: The study comprised nine New Zealand white male rabbits; a rectangular 15 x 5-mm defect was created in the ventral tunica albuginea that was covered by Surgicel. The skin was closed with no catheters left in situ after the procedure. The rabbits were killed in groups of three at 2, 6 and 12 weeks after surgery. The evaluation included cavernosography and histopathological examination of sections of the penis stained with haematoxylin and eosin, and Masson's trichrome. RESULTS: No deaths were caused by the procedure, and none of the rabbits developed bleeding or haematoma after surgery. Cavernosography at 2 weeks showed contrast medium leaking from the site of the Surgicel, but at 6 and 12 weeks all rabbits had a straight erection with patent corpora and no evidence of narrowing or venous leak. Histopathological evaluation revealed evidence of the remnants of Surgicel surrounded by acute inflammatory cell infiltrate with early neovascularization at 2 weeks. At 6 and 12 weeks, there was complete normal regeneration of the tunica albuginea with no foreign-body reaction. CONCLUSIONS: In this pilot study, Surgicel had a clear haemostatic effect when covering a defect in the tunica albuginea. Moreover, normal tunica albuginea regenerated by 6 weeks and was maintained at 12 weeks. These results suggest that Surgicel might be considered a safe and effective grafting material for tunica albuginea substitution, including the surgical management of penile chordee.


Subject(s)
Cellulose, Oxidized/therapeutic use , Hemostatics/therapeutic use , Penile Diseases/surgery , Penis/surgery , Animals , Male , Models, Animal , Penile Erection , Pilot Projects , Rabbits , Treatment Outcome , Wound Healing
18.
J Urol ; 171(5): 1939-42, 2004 May.
Article in English | MEDLINE | ID: mdl-15076316

ABSTRACT

PURPOSE: Previous studies have demonstrated successful use of small intestinal submucosa (SIS) as a tube for replacing short segment (11 mm) proximal ureteral defects. However, such small segment ureteral defects could be managed by resection re-anastomosis. We evaluated the use of 1-layer SIS as a tube for the replacement of long segment ureteral defects. MATERIALS AND METHODS: The ureters of 5 female mongrel dogs were accessed through a median laparotomy incision. A 4 cm segment of mid ureter was resected on the right side. The right ureteral segments were replaced by tubularized SIS segments using 6-zero polydioxanone interrupted sutures. Internal pigtail stents were left for 6 weeks. All animals were sacrificed at 12 weeks. Ureteral patency was assessed by excretory urography and magnetic resonance urography 7 and 12 weeks after the initial procedures. Inflammation and regeneration were assessed histologically. RESULTS: At 12 weeks all ureters on the experimental side were completely occluded with significant hydroureteronephrosis and the subsequent deterioration of kidney function. At autopsy there was failure to calibrate any of the experimental ureters with a 3Fr catheter. Although histologically urothelium and muscular cells had proliferated over the graft, they were embedded in an intense fibrotic and inflammatory process. CONCLUSIONS: Technically 1-layer SIS was easily modeled, providing the conditions for watertight anastomosis. The regeneration of urothelium and muscle was induced and supported by the graft. However, functional replacement was not successful. One-layer SIS is not a suitable material for replacing long segment (4 cm) ureteral defects.


Subject(s)
Intestinal Mucosa/transplantation , Intestine, Small/transplantation , Ureter/surgery , Animals , Dogs , Female , Pilot Projects , Regeneration , Ureter/physiology , Urologic Surgical Procedures/methods
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