Optimization of Phloretin-loaded Nanospanlastics for Targeting of FAS/SREBP1c/AMPK/ OB-Rb Signaling Pathway in HFD-induced Obesity.

OBJECTIVES: Obese patients are at increased risk for CVD, which is the main cause of premature death and has been a major cause of disability and ill health in recent years. PTN, a natural dihydrochalcone flavonoid, has a variety of pharmacological characteristics. This article aimed to prepare PTN-NSLs to evaluate their anti-obesity activity. METHODS: Morphology, Particle size, zeta potential, UV-vis, entrapment efficiency, FT-IR spectra, and an in vitro release study of PTN-NSLs were described. PTN-NSLs were also tested for their anti-obesity properties in obese rats. The LD50 of PTN-NSLs was calculated, as was the 1/20 LD50 prepared for the treatment of obese rats. Also, the level of glycemic, oxidative stress and inflammatory biomarkers were estimated in the obese rat's model. RESULTS: The synthesized PTN-NSLs were uniform, spherically shaped, and well dispersed with no aggregation noted, with a size range of 114.06 ± 8.35 nm. The measured zeta potential value of PTN-NSLs was -32.50.8 mv. Also, the UV spectra of PTN and PTN-NSLs have strong absorption at 225 and 285 nm. Also, the LD50 of PTN-NSLs was found to be 2750 mg/kg.b.w. Moreover, administrating obese rats with PTN-NSLs resulted in improved glycemic features as well as GSH, SOD, GPx, GR, IL10, TBARs, and IL-6 levels, as well as attenuated FAS, SREBP1c, AMPK, ACO, CPT1, and OB-Rb gene expression. CONCLUSIONS: Administration of PTN-NSLs significantly attenuated the levels of glycemic, oxidative stress, and inflammatory biomarkers. The biochemical and PCR findings are aided by histological investigations. Also, the present findings imply that PTN-NSLs might be a promising pharmacological tool for the treatment of obesity-related diseases.

Fully Characterized Effective Bacteriophages Specific against Antibiotic-Resistant Enterococcus faecalis, the Causative Agent of Dental Abscess.

Background and Objectives: Enterococcus faecalis (E. faecalis) is a primary pathogen responsible for dental abscesses, which cause inflammation and pain when trapped between the crown and soft tissues of an erupted tooth. Therefore, this study aims to use specific phages as an alternative method instead of classical treatments based on antibiotics to destroy multidrug-resistant E. faecalis bacteria for treating dental issues. Materials and Methods: In the current study, twenty-five bacterial isolates were obtained from infected dental specimens; only five had the ability to grow on bile esculin agar, and among these five, only two were described to be extensive multidrug-resistant isolates. Results: Two bacterial isolates, Enterococcus faecalis A.R.A.01 [ON797462.1] and Enterococcus faecalis A.R.A.02, were identified biochemically and through 16S rDNA, which were used as hosts for isolating specific phages. Two isolated phages were characterized through TEM imaging, which indicated that E. faecalis_phage-01 had a long and flexible tail, belonging to the family Siphoviridae, while E. faecalis_phage-02 had a contractile tail, belonging to the family Myoviridae. Genetically, two phages were identified through the PCR amplification and sequencing of the RNA ligase of Enterococcus phage vB_EfaS_HEf13, through which our phages shared 97.2% similarity with Enterococcus phage vB-EfaS-HEf13 based on BLAST analysis. Furthermore, through in silico analysis and annotations of the two phages' genomes, it was determined that a total of 69 open reading frames (ORFs) were found to be involved in various functions related to integration excision, replication recombination, repair, stability, and defense. In phage optimization, the two isolated phages exhibited a high specific host range with Enterococcus faecalis among six different bacterial hosts, where E. faecalis_phage-01 had a latent period of 30 min with 115.76 PFU/mL, while E. faecalis_phage-02 had a latent period of 25 min with 80.6 PFU/mL. They were also characterized with stability at wide ranges of pH (4-11) and temperature (10-60 °C), with a low cytotoxic effect on the oral epithelial cell line at different concentrations (1000-31.25 PFU/mL). Conclusions: The findings highlight the promise of phage therapy in dental medicine, offering a novel approach to combating antibiotic resistance and enhancing patient outcomes. Further research and clinical trials will be essential to fully understand the therapeutic potential and safety profile of these bacteriophages in human populations.

Unravelling the Antimicrobial, Antibiofilm, Suppressing Fibronectin Binding Protein A (fnba) and cna Virulence Genes, Anti-Inflammatory and Antioxidant Potential of Biosynthesized Solanum lycopersicum Silver Nanoparticles.

Background and Objectives: Urinary tract infections [UTIs] are considered the third most known risk of infection in human health around the world. There is increasing appreciation for the pathogenicity of Gram-positive and Gram-negative strains in UTIs, aside from fungal infection, as they have numerous virulence factors. Materials and Methods: In this study, fifty urine samples were collected from patients suffering from UTI. Among the isolates of UTI microbes, six isolates were described as MDR isolates after an antibiotic susceptibility test carried out using ten different antibiotics. An alternative treatment for microbial elimination involved the use of biosynthesized silver nanoparticles (AgNPs) derived from Solanum lycopersicum [S. cumin]. Results: The sizes and shapes of AgNPs were characterized through TEM imaging, which showed spherical particles in a size range of 35-80 nm, of which the average size was 53 nm. Additionally, the silver nanoparticles (AgNPs) demonstrated inhibitory activity against Staphylococcus aureus (OR648079), exhibiting a 31 mm zone of inhibition at a minimum inhibitory concentration (MIC) of 4 mg/mL and a minimum bactericidal concentration (MBC) of 8 mg/mL. This was followed by Aspergillus niger (OR648075), which showed a 30 mm inhibition zone at an MIC of 16 mg/mL and a minimum fungicidal concentration (MFC) of 32 mg/mL. Then, Enterococcus faecalis (OR648078), Klebsiella pneumoniae (OR648081), and Acinetobacter baumannii (OR648080) each displayed a 29 mm zone of inhibition at an MIC of 8 mg/mL and an MBC of 16 mg/mL. The least inhibition was observed against Candida auris (OR648076), with a 25 mm inhibition zone at an MIC of 16 mg/mL and an MFC of 32 mg/mL. Furthermore, AgNPs at different concentrations removed DPPH and H2O2 at an IC50 value of 13.54 µg/mL. Also, AgNPs at 3 mg/mL showed remarkable DNA fragmentation in all bacterial strains except Enterococcus faecalis. The phytochemical analysis showed the presence of different active organic components in the plant extract, which concluded that rutin was 88.3 mg/g, garlic acid was 70.4 mg/g, and tannic acid was 23.7 mg/g. Finally, AgNPs concentrations in the range of 3-6 mg/mL showed decreased expression of two of the fundamental genes necessary for biofilm formation within Staphylococcus aureus, fnbA (6 folds), and Cna (12.5 folds) when compared with the RecA gene, which decreased by one-fold when compared with the control sample. These two genes were submitted with NCBI accession numbers [OR682119] and [OR682118], respectively. Conclusions: The findings from this study indicate that biosynthesized AgNPs from Solanum lycopersicum exhibit promising antimicrobial and antioxidant properties against UTI pathogens, including strains resistant to multiple antibiotics. This suggests their potential as an effective alternative treatment for UTIs. Further research is warranted to fully understand the mechanisms of action and to explore the therapeutic applications of these nanoparticles in combating UTIs.

A New Proposed Combined CT and MRI Staging System for Covid-19-Associated Rhino-Orbito-Cerebral Fungal Infection: A Multi-center Study with Pathological Correlation.

RATIONALE AND OBJECTIVES: To propose a combined computed tomography (CT) and magnetic resonance imaging (MRI) based classification system in the management of COVID-19-associated rhino-orbito-cerebral (C-ROC) fungal infection and to assess the reliability of such proposed staging system. MATERIALS AND METHODS: This was a multi-center prospective study conducted on 122 adults with previously confirmed COVID-19 infection. CT and contrast-enhanced MRI (CE-MRI) were performed for all patients. Three radiologists (with experience of 8, 10, and 14 years) independently assessed all images. Then, each patient was assigned a radiological stage based on the five stages of the proposed system according to the radiological extent of the fungal infection. The intra-class correlation coefficient (ICC) test assessed the inter-rater agreement. Based on the pathological evaluation of post-operative specimens, a diagnosis of fungal infection was documented. RESULTS: The most prevalent severity stage among all raters was stage IV in 29.5-31.1% patients. The overall inter-rater agreement of the proposed staging system was excellent (ICC 0.971, 95% CI;0.960-0.979). Moreover, the most common detected pathogen was Mucormycosis (n = 87, 71.3%). Furthermore, there was a statistically significant association between the patients' outcome and the severity stage (P value 0.001) and there was no statistically significant association between ethmoid and sphenoid sinus affection and cranial extension (P value 0.081). CONCLUSION: Our proposed combined CT and MRI severity staging system has a high inter-rater agreement. Moreover, it can aid in the early detection of the C-ROC fungal infection, improve preoperative planning, and subsequently improve the patient's outcome.

FolE gene expression for folic acid productivity from optimized and characterized probiotic Lactobacillus delbrueckii.

BACKGROUND: Lactobacillus delbrueckii was one of the most common milk lactic acid bacterial strains (LAB) which characterized as probiotic with many health influencing properties. RESULTS: Among seven isolates, KH1 isolate was the best producer of folic acid with 100 µg/ml after 48 h of incubation; FolE gene expression after 24 h of incubation was in the highest value in case of KH1 with three folds. Lactose was the best carbon source for this KH1, besides the best next isolates KH80 and KH98. The selected three LAB isolates were identified through 16S rDNA as Lactobacillus delbrueckii. These three isolates have high tolerance against acidic pH 2-3; they give 45, 10, and 22 CFUs at pH 3, besides 9, 6, and 4 CFUs at pH2, respectively. They also have resistance against elevated bile salt range 0.1-0.4%. KH1 recorded 99% scavenging against 97.3% 1000 µg/ml ascorbic acid. Docking study exhibits the binding mode of folic acid which exhibited an energy binding of - 8.65 kcal/mol against DHFR. Folic acid formed four Pi-alkyl, Pi-Pi, and Pi-sigma interactions with Ala9, Ile7, Phe34, and Ile60. Additionally, folic acid interacted with Glu30 and Asn64 by three hydrogen bonds with 1.77, 1.76, and 1.96 Å. CONCLUSION: LAB isolates have probiotic properties, antioxidant activity, and desired organic natural source for folic acid supplementation that improve hemoglobin that indicated by docking study interaction.

Green synthesis of silver nanoparticles derived from lemon and pomegranate peel extracts to combat multidrug-resistant bacterial isolates.

BACKGROUND: Multidrug-resistant (MDR) bacteria are acknowledged as one of the main factors contributing to chronic illnesses and fatalities globally. Numerous diseases, including bloodstream infections, pneumonia, urinary tract infections, and surgical site infections, can be brought on by MDR bacteria. Therefore, a crucial topic of continuing research is the development of a novel and different treatment for MDR microbial pathogens. This work is introduce an alternative method for elimination of MDR bacterial isolates which are causative agents of urinary tract infection among people in Egypt. In our study, we need a novel strategy to combat MDR bacteria by green-synthesized metal nanoparticles (MNPs). That is due to the ability of MNPs to penetrate the cell wall and the cell membrane of gram-positive and gram-negative bacteria. METHODS: Clinical isolates of MDR bacteria had their antibiotic susceptibility assessed before being molecularly identified using 16 s rRNA, sequencing, and phylogenetic analysis. Also, genetic profiles of isolated strains were performed using ISSR and SDS-PAGE. Finally, characterized plant-mediated silver nanoparticles derived from lemon and pomegranate peel extracts were evaluated against isolated multidrug-resistant bacterial stains. RESULTS: In our present trial, one-hundred urine samples were collected from 71 females and 29 males complaining of UTI (urinary tract infection) symptoms. One-hundred microbial isolates were isolated, including 88-g negative and only 8-g positive bacteria in addition to four yeast isolates (Candida species). A total of 72% of the isolated bacteria showed MDR activity. The most prevalent MDR bacterial isolates (Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterococcus faecalis, and Klebsiella pneumoniae) were identified through 16S rDNA PCR sequencing as with accession numbers OP741103, OP741104, OP741105, OP741106, and OP741107, respectively. Lemon and pomegranate-mediated silver nanoparticles [Ag-NPs] were characterized by UV spectroscopy, FTIR, XRD, and TEM with average size 32 and 28 nm, respectively. Lemon and pomegranate-mediated silver nanoparticles [Ag-NPs] showed an inhibitory effect on the selected five MDR isolates at MIC 50 and 30 µg/mL, respectively. These common bacterial isolates were also genetically examined using ISSR PCR, and their total protein level was evaluated using SDS-PAGE, showing the presence of distinct genetic and protein bands for each bacterial species and emphasizing their general and protein composition as a crucial and essential tool in understanding and overcoming MDR behavior in UTI patients. CONCLUSIONS: Lemon and pomegranate-mediated silver nanoparticles [Ag-NPs] were found to have an inhibitory effect on MDR isolates. Therefore, the study suggests that [Ag-NPs] could be a potential treatment for MDR UTI infections caused by the identified bacterial species.

Versatility of Local Fasciocutaneous Island Flaps for Resurfacing Soft Tissue Defects Overlying the Achilles Tendon.

Background: Reconstruction of soft tissue defects overlying the Achilles tendon has always been a challenge. Various modalities of reconstruction have been described to resurface such defects. We aimed to assess the functional and cosmetic outcomes of all patients who had undergone reconstruction of small and medium sized soft tissue defects of the Achilles region using local fasciocutaneous island flaps. Methods: This retrospective study was conducted from January 2020 to June 2022. 15 patients with small (≤ 30 cm2) and medium (30-90 cm2) sized soft tissue defects of the tendo-Achilles region, underwent reconstruction with local fasciocutaneous island flaps and had complete medical records, were included. Results: Thirteen patients were male (86.7%). The mean age was 53.2 years. 5 cases (33.3%) had post-traumatic open AT injuries with skin avulsion, while ten patients (66.7%) had suture line complications after open repair of spontaneous Achilles tendon rupture. Defect sizes ranged from 12 to 63 cm2. Reverse sural flap was used in 5 patients (33.3%) and medial plantar flap in 10 patients (66.7%). All flaps survived completely. Complications were detected in 3 patients (20%); 1 distal superficial necrosis in a sural flap and 2 marginal minimal graft loss. Functional outcome was good in 12 patients (80%), excellent in 1 patient (6.7%) and fair in 2 patients (13.3%). 13 patients (86.7%) were satisfied with the cosmetic results. Conclusion: Local fasciocutenous island flaps are reliable and simple solutions for covering small to moderate soft tissue defects overlying the Achilles Tendon, with acceptable functional and cosmetic outcomes.

A Novel Approach of Periodontal Osseous Wall Piezosplitting and Sequential Bone Expansion in Management of Localized Intra-Bony Defects with Wide Angulation-A Randomized Controlled Trial.

Piezoelectric surgical instruments with various mini-sized tips and cutting technology offer a precise and thin cutting line that could allow the wider use of periodontal osseous wall swaging. This randomized controlled trial was designed to investigate the use of a minimally invasive piezo knife to harvest vascularized interseptal bone pedicles in treating intra-bony defects. Sixteen non-smoking patients (mean age 39.6 ± 3.9) with severe chronic periodontitis were randomly assigned into one of two groups (N = 8). The Group 1 (control) patients were treated by bone substitute grafting of the intra-bony defect, whereas the Group 2 patients were treated by intra-bony defect osseous wall swaging (OWS) combined with xenograft filling of the space created by bone tilting. In both groups, the root surfaces were treated with a neutral 24% EDTA gel followed by saline irrigation. Clinical and radiographic measurements were obtained at baseline and 6 months after surgery. The sites treated with osseous wall swaging showed a statistically significant probing-depth reduction and increase in clinical attachment compared with those of the Group 1 patients. The defect base level was significantly reduced for the OWS group compared to that of the Group 1 control. By contrast, the crestal bone level was significantly higher in the OWS group compared to Group 1. The crestal interseptal bone width was significantly higher in Group 2 at 6 months compared to the baseline value and to that of Group 1 (<0.001). The osseous wall swaging effectively improved the clinical hard- and soft-tissue parameters. The use of mini inserts piezo-cutting, sequential bone expanders for osseous wall redirection, and root surface EDTA etching appears to be a reliable approach that could allow the use of OWS at any interproximal dimension.

Elimination of pathogenic multidrug resistant isolates through different metal oxide nanoparticles synthesized from organic plant and microbial sources.

Beta-hemolytic multidrug-resistant bacteria (MDR) are highly regarded as a major public health risk because they are resistant to at least 10 antibiotics in different groups with different mechanisms of action. The present study shows that among 98 bacterial isolates collected from laboratory fecal samples: 15 were beta-hemolytic and tested against 10 different antibiotics. 15 beta-hemolytic; 5 isolates exhibit strong multidrug resistance traits. Isolate 5 Escherichia coli (E. coli), Isolate 7 (E. coli), Isolate 21 (Enterococcus faecium), Isolate 27 (Staphylococcus sciuri), and isolate 36 (E. coli) are largely untested antibiotics. Substances (clear zone >10 mm) Its growth sensitivity to different types of nanoparticles was further evaluated by the agar well diffusion method. AgO, TiO2, ZnO, and Fe3O4 nanoparticles have been separately synthesized by microbial and plant-mediated biosynthesis. By evaluating the antibacterial activity of different nanoparticle types against selected MDR isolates, the results showed that global MDR bacterial growth was inhibited differently depending on the nanoparticle type. TiO2 was the most potent antibacterial nanoparticle type, followed by AgO, while Fe3O4 showed the least efficacy against selected isolates. The MICs of microbially synthesized AgO and TiO2 nanoparticles were 3 µg (67.2 µg/mL) and 9 µg (180 µg/mL) for isolates 5 and 27, respectively, indicating that biosynthetic nanoparticles via pomegranate of antibacterial activity at a higher MIC than microbial-mediated ones, it recorded (300 and 375 µg/ml, respectively) of AgO and TiO2 nanoparticles for isolates 5 and 27. Biosynthesized nanoparticles were examined by TEM, the average sizes of microbial AgO and TiO2 nanoparticles were 30 nm and 70 nm, respectively, and the average sizes of plant mediated AgO and TiO2 NPs were 52 nm and 82 nm respectively. Two most potent extensive MDR isolates (5 and 27) were identified as E. coli and Staphylococcus sciuri by 16s rDNA technology, and the sequencing results of the isolates were deposited with NCBI GenBank under accession numbers ON739202 and ON739204, respectively.

Utilization of Non-Drug-Eluting Devices for Inpatient Percutaneous Coronary Intervention in the United States.

There is a paucity of data on the contemporary use of non-drug-eluting devices (balloon angioplasty or bare-metal stents) in contemporary percutaneous coronary intervention (PCI) in the United States. We utilized the Nationwide Readmissions Database to identify patients hospitalized to undergo PCI with non-drug-eluting devices from 2016 to 2019. The main outcome of interest was the trends in utilization over the study years. Among 1,870,262 PCI procedures, 127,810 (6.8%) were performed with non-drug-eluting devices; 72% of these were in the setting of acute myocardial infarction (MI). The use of non-drug-eluting devices decreased throughout the study period from 12.9% of all PCI in the first quarter of 2016 to 3.4% in the last quarter of 2019 (p <0.001). Factors associated with their use included advanced age and high bleeding risk. Only a small percentage were used as a bridge to coronary artery bypass graft surgery (2%) and for treatment of in-stent restenosis (3%). The in-hospital mortality was 5.8% for the entire cohort and 6.6% when the indication for use was an acute MI. In patients presenting with an acute MI, reinfarction within 30 days was common and occurred in 18% of the patients. In conclusion, the use of non-drug-eluting devices in PCI in the United States decreased from 2016 to 2019. Factors associated with their use included old age and high bleeding risk. Due to suboptimal outcomes in patients currently being treated with non-drug-eluting devices, there remains an unmet clinical need for alternative treatment options.

Novel Cold Cure Acrylic Denture Base with Recycled Zirconia Nano-Fillers That Were Functionalized by HEMA Agent Incorporation: Using the Sprinkle Approach.

Background: Though acrylic resins possess many useful properties, denture fracture is nevertheless a familiar issue. Objective: This study aimed to determine the effect of low-percent recycled Zirconia nanoparticles as filler on the transverse strength, impact strength, surface hardness, water sorption, and solubility of resin using the sprinkle cold-curing technique. Materials and Methods: Various formulae were prepared and mixed with PMMA (polymer) powder containing varying percentages (0.01%, 0.1%, 0.3%, and 0.5%) of recycled ZrO2NPs to mono-methyl methacrylate (MMA monomer). A 2-hydroxyethyl-methacrylate (HEMA) agent was used to functionalize recycled zirconia (ZrO2) nano-fillers. X-ray diffraction, field emission scanning electron microscopy, high-resolution transmission electron microscopy, energy-dispersive X-ray spectroscopy, and dynamic light scattering were used to characterize the samples. For mechanical tests, standard metallic moulds (according to American Dental Association specification no. 27) were machined for 60 specimens' preparation, 12 for each percent (zero, 0.01%, 0.1%, 0.3%, and 0.5%). A one-way ANOVA test was used to compare the five groups for parametric data, while the Kruskal-Wallis test was employed for nonparametric data. The P 0.05 value was accepted as the significance level. All formulae were tested for cytotoxicity at 24 and 48 hours on WI38 normal lung cell lines. Results: The XRD analysis demonstrated the tetragonal crystallographic structure of the recycled zirconia nanoparticles. Incorporating a low percentage of recycled ZrO2 nanoparticles (0.01%, 0.1%, 0.3%, and 0.5%) improved the tested properties of PMMA to different degrees in a significant and non-significant pattern, while the optimal tested percent was 0.3%. Conclusion: The 0.3% percentage of recycled zirconia nanoparticles maintained and improved the physical and mechanical properties of acrylic resin. Recycled ZrO2/PMMA nanocomposite is a synergistic candidate due to its economic return and clinical application safety.

Molecular docking and nucleotide sequencing of successive expressed recombinant fungal peroxidase gene in E.coli.

BACKGROUND: Fungal peroxidases are oxidoreductases that utilize hydrogen peroxide to catalyze lignin biodegradation. RESULTS: PER-K (peroxidase synthesis codon gene) was transformed from Aspergillus niger strain AN512 deposited in the National Center for Biotechnology Information with the accession number OK323140 to Escherichia coli strain (BL21-T7 with YEp356R recombinant plasmid) via calcium chloride heat-shock method. The impact of four parameters (CaCl2 concentrations, centrifugation time, shaking speed, growth intensity) on the efficacy of the transformation process was evaluated. Furthermore, peroxidase production after optimization was assessed both qualitatively and quantitatively, as well as SDS-PAGE analysis. The optimum conditions for a successful transformation process were as follows: CaCl2 concentrations (50 mM), centrifugation time (20 min), shaking speed (200 rpm), and growth optical density (0.45). PCR and gel electrophoresis detect DNA bands with lengths 175, 179, and 211 bps corresponding to UA3, AmpR, and PER-K genes respectively besides partially sequencing the PER-K gene. Pyrogallol/hydrogen peroxide assay confirmed peroxidase production, and the activity of the enzyme was determined to be 3924 U/L. SDS-PAGE analysis also confirms peroxidase production illustrated by the appearance of a single peroxidase protein band after staining with Coomassie blue R-250. CONCLUSION: A successful peroxidase-gene (PER-K) transformation from fungi to bacteria was performed correctly. The enzyme activity was screened, and partial sequencing of PER-K gene was analyzed successively. The protein 3D structure was generated via in silico homology modeling, and determination of binding sites and biological annotations of the constructed protein were carried out via COACH and COFACTOR based on the I-TASSER structure prediction.

Aorta unfolding index as an indicator for coronary artery disease severity.

BACKGROUND: Aortic unfolding index (AUI) is an independent predictor of cardiovascular events, yet there is scarcity in the literature on its association with the severity of coronary artery disease (CAD). PURPOSE: To investigate the correlation between aortic unfolding and coronary artery disease severity score. MATERIAL AND METHODS: The study included 115 patients with various degrees of CAD who underwent invasive coronary angiography and were retrospectively studied. AUI derived from non-contrast computed tomography (CT) of the chest was correlated to the Gensini score describing the CAD severity. Its sensitivity and specificity in the detection of severe stenosis were examined at various cutoff points. RESULTS: CAD severity was significantly correlated with the patient age and AUI. On multivariate regression analysis, AUI was an independent predictor of severe CAD. The best cutoff value was ≥66, with 94.9% sensitivity and 81.6% specificity. CONCLUSION: AUI ≥66 was a predictor of severe CAD independent of the patient age.

Occurrence and clinical characteristics of occult hepatitis C virus infection in hemodialysis units at Ismailia, Egypt.

BACKGROUND AND AIMS: Occult hepatitis C infection is defined as the presence of hepatitis C virus RNA in peripheral blood mononuclear cells (PBMC) ± hepatocytes in the absence of HCV-RNA in serum. It has been a lot of debate and controversy in recent years and not discussed well. This issue has not been discussed or investigated in Egypt, especially in patients on hemodialysis in Ismailia. This study is the first one to investigate the prevalence of occult HCV infection in large populations of chronic hemodialysis (CHD) patients in Ismailia, Egypt. METHODS: Our study is cross-sectional analytic and included 204 CHD patients; who are negative for HCV infection. Sensitive commercial real-time assay was used to detect HCV-RNA in PBMC. In our study, the presence of genomic HCV-RNA in PBMCs of all these patients was detected by real-time PCR. On the other hand, 22 patients on hemodialysis with an established diagnosis of chronic hepatitis C virus infection were included as a control group and examined by real-time PCR was used to evaluate HCV infection. RESULTS: Occult HCV infection is defined as the presence of HCV-RNA in PBMNCs in patients on chronic hemodialysis, and it was found in 14/204 (7%) of the patients. Patients who were on CHD for a longer time are susceptible to occult HCV infection, and their mean alanine aminotransferase levels are significantly higher during the last 3 months before study entry. In comparison, chronic HCV patients have elevated bilirubin, aspartate transferase and alanine transferase than occult HCV infection. CONCLUSIONS: The prevalence of occult HCV infection was 7% in our CHD patients. No available data are showing the virulence of this form of virus. However, further studies in other geographic populations with high HCV endemicity are needed to clarify the significance of occult HCV infection in these patient groups, in addition to test for the presence of negative antigenomic strand to confirm or disconfirm the reliability of occult HCV.

Recombinant overexpression of the Escherichia coli acetyl-CoA carboxylase gene in Synechocystis sp. boosts lipid production.

Microalgae have received continued attention as a potential source for biofuel production. However, the lack of suitable strains that provide a lipid-rich biomass and tolerate harsh condition inhibits their industrial application. This report describes an effort to transform Synechocystis sp. with genes encoding acetyl-CoA carboxylase (ACC), a key regulatory enzyme in the lipogenesis pathway, from the white mustard plant (Sinapis alba) and the bacterium Escherichia coli DH5α using chitosan nanoparticles. Although a recombinant plasmid encoding S. alba ACC failed to express, successful transformation was achieved with a recombinant plasmid encoding E. coli DH5α ACC. The successful transformant, Synechocystis sp. PAK13, exhibited increased ACC expression compared with its wild-type parent (11.8 vs. 7.2 ng), which significantly increased its lipid content (by 3.6-fold). Synechocystis sp. PAK13 also exhibited a significant (20%) reduction in photosynthetic pigments, a 1.52-fold higher glucose content and a 3.5-fold lower sucrose content than the wild-type. In conclusion, this report introduces a useful strategy to overexpress the ACC gene in microalgae, creating strains with improved lipid production that are suited to industrial applications.

Spatiotemporal Characteristics and Driving Factors of Black Carbon in Augsburg, Germany: Combination of Mobile Monitoring and Street View Images.

The study investigates the spatial pattern of black carbon (BC) at a high spatial resolution in Augsburg, Germany. Sixty two walks were performed to assess the concentrations of equivalent black carbon (eBC), ultraviolet particulate matter (UVPM), and equivalent brown carbon (eBrC) in different seasons and at different times of the day with a mobile platform (i.e., trolley). Along with BC measurements, images of street microenvironments were recorded. Meteorological parameters, including temperature, relative humidity, and wind speed, were monitored. The BC concentrations showed significant spatial heterogeneity and diurnal variations peaking in the morning and at night. The highest BC concentrations were observed near dense traffic. The correlations between BC and street views (buildings, roads, cars, and vegetation) were weak but highly significant. Moreover, meteorological factors also influenced the BC concentration. A model based on street view images and meteorological data was developed to examine the driving factors of the spatial variability of BC concentrations at a higher spatial resolution as different microenvironments based on traffic density. The best results were obtained for UVPM and eBC (71 and 70% explained variability). eBrC (53%), to which other sources besides road traffic can also make significant contributions, is modeled less well.

Autophagy blockade mechanistically links proton pump inhibitors to worsened diabetic nephropathy and aborts the renoprotection of metformin/enalapril.

AIM: Proton pump inhibitors (PPIs) are widely used drugs recently linked to chronic kidney disease. However, the invloved mechanisms remained elusive. Since defective autophagy is identified as a new culprit in the pathogenesis of diabetic nephropathy (DN), we aimed to trace the link of autophagy blockade by PPIs to the progression of DN with and without the standard therapy of metformin and enalapril. MAIN METHODS: Male CD1 albino mice (20-25 g) were randomly assigned to normal control or diabetic mice. Diabetes was induced by intraperitoneal streptozotocin (35 mg/kg) injection combined with high fat diet. DN mice were randomized to receive vehicle, lansoprazole (5 mg/kg), metformin (200 mg/kg), lansoprazole + metformin, metformin + enalapril (0.5 mg/kg) or the three drugs together, orally daily for four weeks. At the study end, albuminuria, fasting plasma glucose, HbA1c, renal functions and malondialdehyde were assessed. Renal tissues were examined microscopically, and autophagic changes were evaluated by immunohistochemical detection of LC3-II and p62. KEY FINDINGS: Consistent with autophagic blockade, lansoprazole increased both LC3II and p62 in the glomerular and tubular cells. This was associated with impaired creatinine clearance and renal functions, enhanced albuminuria, oxidative stress and augmented DN histopathological changes. Opposite effects on autophagy markers were observed by single or combined treatment of metformin with enalapril; which also ameliorated glycemic control and signs of DN. This improvement was mitigated by combination with lansoprazole. SIGNIFICANCE: Autophagy blockade by lansoprazole augmented diabetic nephropathy and opposed the reno-protective effects of metformin and enalapril. The use of PPIs in diabetes should be considered with great caution.

Improving wear resistance of plasma-sprayed calcia and magnesia-stabilized zirconia mixed coating: roles of phase stability and microstructure.

The phase stability and microstructure of ZrO2-5CaO and ZrO2-24MgO mixed coating (wt%) by air plasma spraying on 304 stainless steel substrates were investigated. A Ni-5Al (wt%) metallic bond coating was firstly sprayed between the substrate and the ceramic top layer. The results were compared with the individual coatings of ZrO2-5CaO and ZrO2-24MgO for a better understanding of the correlation between their microstructures and mechanical properties. Mixed zirconia coating was found to have a mixture of cubic and tetragonal phases that stabilized under different plasma spray conditions. Microscopic observations and elemental composition analysis of as-sprayed mixed coating showed that modified ceramic-matrix grains had been formed. Microsized ZrO2-5CaO particles were embedded in the matrix grain creating an intragranular microstructure. Results indicated that ceramic-matrix grains provided a diffusion barrier for the growth of oxides induced stress near and onto the bond layer that reduced cracks, thereby overcoming the top delamination of the ceramic coating. Moreover, disparity in wear resistance and microhardness behavior of the coatings was influenced by initial feedstock powder and matrix microstructures. Improvement in the wear resistance of the mixed zirconia coating was attributed to a decrease in oxide content, which resulted in an increase in intersplat cohesive strength.

Activation of LacZ gene in Escherichia coli DH5α via α-complementation mechanism for ß-galactosidase production and its biochemical characterizations.

BACKGROUND: Plasmid propagation in recombination strains such as Escherichia coli DH5α is regarded as a beneficial instrument for stable amplification of the DNA materials. Here, we show trans-conjugation of pGEM-T cloning vector (modified Promega PCR product cloning vector with tra genes, transposable element (Tn5)) and M13 sequence via α-complementation mechanism in order to activate ß-D-galactosidase gene in DH5α strain (non-lactose-fermenting host). RESULTS: Trans-conjugation with pGEM-T allows correction of LacZ gene deletion through Tn5, and successful trans-conjugants in DH5α host cells can be able to produce active enzyme, thus described as lactose fermenting strain. The intracellular ß-galactosidase was subjected to precipitation by ammonium sulfate and subsequently gel filtration, and the purified enzyme showed a molecular weight of approximately 72-kDa sodium dodecyl sulfate-polyacrylamid gel electrophoresis. The purified enzyme activity showed an optimal pH and temperature of 7.5 and 40 °C, respectively; it had a high stability within pH 6-8.5 and moderate thermal stability up to 50 °C. CONCLUSION: Trans-conjugant of E. coli DH5α- lacZ∆M15 was successfully implemented. UV mutagenesis of the potent trans-conjugant isolate provides an improvement of the enzyme productivity. The enzymatic competitive inhibition by D-galactose and hydrolysis of lactose at ambient temperatures could make this enzyme a promising candidate for use in the dairy industry.