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Klebsiella pneumoniae is a major drug-resistant human pathogen accountable for a wide range of infections. In this cross-sectional study, we aimed to determine the phenotypic and genotypic features of ß-lactamase-producing K. pneumoniae clinical isolates from Alexandria, Egypt. A total of 50 nonduplicated clinical isolates of K. pneumoniae were obtained from various specimens. They were identified biochemically and by biotyping using mass spectrometry. For molecular characterization, plasmid profile analysis was performed. Screening for extended spectrum ß-lactamases (ESBLs), carbapenemases and AmpC production was carried out phenotypically and genotypically. Correlation analysis was performed to assess the relationship between phenotype, genotype and resistance patterns among the studied isolates. The dendrogram demonstrated 38 distinct plasmid profiles among 62% of our isolates. According to antimicrobial susceptibility testing, 90% of isolates were multi/extensive-drug resistant. Nineteen out of 50 (38%) were resistant to cefoxitin, while only 10 (20%) were resistant to imipenem. All isolates were susceptible to colistin. Phenotypically, ESBL producers (78%) were the most common, followed by carbapenemase producers (24%). Genotypically, the most common ESBL gene was blaSHV (90%), followed by blaCTX-Mu (74%), while the most common carbapenemase genes were blaNDM (56%) and blaOXA-48 (54%). No blaKPC or blaIMP were detected. Plasmid-mediated AmpC resistance was confirmed in only two out of 19 cefoxitin-resistant isolates. Both the blaNDM and blaOXA.48 genes were significantly positive correlated (rho = 0.56, p = 0.004). Absence of blaKPC among carbapenem resistant K. pneumoniae isolates in Alexandria, Egypt. AmpC production is not the main factor behind the resistance to cefoxitin among our isolates.
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BACKGROUND AND OBJECTIVES: Immunocompromised patients are a high-risk group for developing mycobacterial infections with either pulmonary and/or extrapulmonary diseases. Low-cost/density DNA-microarray is considered an easy and efficient method for the detection of typical and atypical mycobacterial species. MATERIALS AND METHODS: Thirty immunocompromised patients were recruited to provide their clinical specimens (sputum, serum, urine, and lymph node aspirates). Real-time polymerase chain reaction (PCR) and DNA-microarray techniques were performed and compared to the conventional methods of Ziehl-Neelsen staining and Lowenstein Jensen culturing. RESULTS: Mycobacterium tuberculosis complex was detected in all 30 clinical specimens (100% sensitivity) by real-time PCR and DNA-microarray. Additionally, coinfection with 4 atypical species belonging to nontuberculous mycobacteria was identified in 7 sputum specimens. These atypical mycobacterial species were identified as M. kansasii 10% (n = 3), M. avium complex 6.6% (n = 2), M. gordanae 3.3% (n = 1), and M. peregrinum 3.3% (n = 1). CONCLUSION: This study documents the presence of certain species of atypical mycobacteria among immunocompromised patients in Egypt.
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Mycobacterium tuberculosis , Tuberculose , DNA , Egito , Humanos , Mycobacterium tuberculosis/genética , Micobactérias não Tuberculosas/genética , Escarro/microbiologia , Tuberculose/diagnóstico , Tuberculose/microbiologiaRESUMO
BACKGROUND: Chemical Plaque control by antimicrobial agent application can defend the teeth against caries. S. mutans is considered the main etiologic factor for caries. This was an in vitro study to compare between the efficacy of chlorhexidine diaceteate varnish, and an organoselenium sealant, to prevent S. mutans biofilm formation on human teeth. METHODS: Fourty five premolars extracted for orthodontic purposes were randomly divided into 3 groups of 15 teeth each. One control group and two test groups, chlorhexidine diaceteate varnish and an organoselenium sealant. The teeth were autoclaved before S. mutans biofilm was induced on to each in their respective groups. The reading T1 was taken for each tooth to assess the number of S. mutans attached in order to compare for differences in surface area among the 3 groups. The respective test materials were applied onto the teeth and biofilm induced onto them in their respective groups. The reading T2 was taken for the 2 test groups. The 3 groups were then subjected to aging for a period equivalent to 5 months before the biofilm was induced to take the reading T3 for the number of S. mutans. We used vortexing of the teeth to disrupt the biofilm at time points T1, T2 and T3. S. mutans count was then done using PCR. RESULTS: There were significantly lower S. mutans counts in the control group as compared to the chlorhexidine diacetate group at T3.There were no other statistically significant differences found. CONCLUSION: Both organoselenium and Chlorhexidine diacetate do not inhibit S. mutans biofilm attachment onto the teeth.
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Anti-Infecciosos Locais , Cárie Dentária , Placa Dentária , Anti-Infecciosos Locais/farmacologia , Biofilmes , Clorexidina/farmacologia , Contagem de Colônia Microbiana , Cárie Dentária/prevenção & controle , Placa Dentária/prevenção & controle , Humanos , Streptococcus mutansRESUMO
Direct acting antiviral agents (DAAs) are a group of antiviral drugs that inhibit specific non-structural proteins of the virus and disrupt viral replication and infection. DAAs regimens for hepatitis C virus (HCV) infection provide a particular event to tackle mechanistic intracellular relationships between the innate immunity and HCV, potentially providing perceptions about the rate of the viral replication and complex decay. Interleukin 29 (IL-29) prevents the replication of HCV. IFN-inducible protein 10 (IP-10) plays a significant role in the pathogenesis of HCV infection. MIG/CXCL9 are produced by inï¬ammatory and stromal cells such as hepatocytes following either stimulation by interferon lambda (IFNγ) or viral infection. This study aimed to evaluate the co-expression of IL-29, IP-10 and MIG in peripheral blood mononuclear cells (PBMCs) from untreated and treated chronic HCV patients with DAAs. This study included group of twenty naïve HCV patients, group of twenty sustained viral response (SVR) patients and a control group that consisted of 10 healthy subjects. All subjects were tested for liver enzymes, serum albumin level, total serum bilirubin, platelet count, prothrombin activity and viral load. Relative gene expression of IL-29, IP-10, and MIG in PBMCs from all subjects was determined using real time PCR. The mean value of IL-29, IP-10 and MIG gene expression significantly increased in both naïve HCV and SVR groups of patients as compared to normal subjects. The corresponding value was significantly lower in patients with SVR compared to naïve HCV patients. Infection with HCV significantly trigged the co-expression of IL-29, IP-10, and CXCL9 (MIG) genes in PBMCs of chronic hepatitis C patients and significantly down-regulated in those who achieved SVR after successful DAAs therapy. Keywords: IP10; MIG; IL29; HCV; DAAs; gene expression.
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Hepatite C Crônica , Antivirais/uso terapêutico , Quimiocina CXCL10/genética , Egito , Hepacivirus , Hepatite C Crônica/tratamento farmacológico , Hepatite C Crônica/genética , Humanos , Interleucinas/genética , Leucócitos Mononucleares , Monocinas/uso terapêuticoRESUMO
Emerging hepatocellular carcinoma (HCC) has been sequentially reported in chronic hepatitis C virus (HCV) treated with direct-acting antivirals (DAAs). Homeobox transcript antisense RNA (HOTAIR), an oncogene, has been reported to be associated with cancer. We investigated the predictive value of lnc-HOTAIR for HCC surveillance in chronic HCV patients following DAAs therapy. The expression levels of lnc-HOTAIR and ATG-7 genes were measured in 220 with chronic HCV, following a DAAs based therapy for 12 weeks, the patients were followed-up for attentive surveillance of HCC for 12 months after starting DAAs. In terms of lnc-HOTAIR, patients with HCC and high viral load had significantly higher median expression levels of HOTAIR of (68 vs. 24; p = .001) and (94 vs. 52; p = .001), respectively. Moreover, the median expression level of ATG-7 was higher in those who developed HCC (114 vs. 51; p = .001). The expression of lnc-HOTAIR and ATG-7 are significant predictors of the development of HCC in HCV-4 infected patients treated with DAAs, with a cut-off value of 37 and 86, respectively. The increased expression levels of lnc-HOTAIR more than 68 in HCC patients following DAAs were correlated with poorer disease outcomes compared to those with lower expression levels; however, ATG-7 expression levels more than 114 were correlated with worse overall survival but not the progression-free one. We suggest that high expression levels of lnc-HOTAIR could serve as a risk assessment biomarker for HCC before and during DAAs course therapy in Chronic HCV-4 patients, and should be rigorously taken into consideration before DAAs.
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Antivirais/administração & dosagem , Proteína 7 Relacionada à Autofagia/genética , Carcinoma Hepatocelular/virologia , Hepacivirus/genética , Hepatite C Crônica/tratamento farmacológico , Neoplasias Hepáticas/virologia , RNA Longo não Codificante/genética , Idoso , Antivirais/farmacologia , Proteína 7 Relacionada à Autofagia/metabolismo , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Progressão da Doença , Feminino , Genótipo , Hepacivirus/efeitos dos fármacos , Hepacivirus/crescimento & desenvolvimento , Hepatite C Crônica/complicações , Hepatite C Crônica/genética , Hepatite C Crônica/metabolismo , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Masculino , Pessoa de Meia-Idade , Prognóstico , Análise de Sobrevida , Resultado do Tratamento , Regulação para Cima , Carga Viral/efeitos dos fármacosRESUMO
Breast cancer is the second most common cancer diagnosed worldwide. Human epidermal growth factor receptor 2 (HER2)-positive breast cancer represents about 20% to 30% of all breast cancers. Trastuzumab is used in the treatment of HER2-positive breast cancer. MicroRNA-21 (miR-21) is an oncomiR that acts by inhibiting many tumor-suppressor genes. We analyzed the relative expression levels of serum miR-21 in 20 HER2-positive metastatic breast cancer patients before and after 3 months of treatment with trastuzumab. miR-21 levels decreased with a high significant difference after trastuzumab therapy (P = 0.001). Although miR-21 expression levels were lower in responders than in nonresponders, the difference was not statistically significant ( P = 0.6). Our results demonstrated a significant negative correlation between its basal expression, expression levels after treatment, and time to progression ( P = 0.03 and 0.01, respectively). These results make miR-21 a potential prognostic factor for HER2-positive metastatic breast cancer patients. Additionally, it can be an interesting potential target in therapy using antisense oligonucleotides for miR-21.
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Antineoplásicos Imunológicos/uso terapêutico , Biomarcadores Tumorais/genética , Neoplasias da Mama/patologia , MicroRNAs/genética , Receptor ErbB-2/metabolismo , Trastuzumab/uso terapêutico , Adulto , Idoso , Biomarcadores Tumorais/sangue , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Feminino , Seguimentos , Regulação Neoplásica da Expressão Gênica , Humanos , MicroRNAs/sangue , Pessoa de Meia-Idade , Metástase Neoplásica , Prognóstico , Taxa de SobrevidaRESUMO
PURPOSE: Plasmacytoid dendritic cells (PDCs) infiltration into breast cancer tissues is associated with poor prognosis. Also, CXCR4 shows compelling evidences to be exploited by cancer cells to migrate to distant sites. The present study investigated lymph node metastasis in the light of PDCs infiltration and the potential cross talk with CXCR4/SDF-1 chemokine axis. METHODS: We assessed circulating PDCs proportions drained from the axillary tributaries, and the in situ expression of both CD303 and CXCR4 in breast cancer patients with positive lymph nodes (pLN) and negative lymph nodes (nLN) using immunohistochemistry and flow cytometry. We also analyzed the expression of SDF-1 in lymph nodes of pLN and nLN patients. We studied the effect of the secretome of PDCs of pLN and nLN patients on the expression of CXCR4 and activation of NF-κB in human breast cancer cell lines SKBR3 and MCF-7. TNF-α mRNA expression level in PDCs from both groups was determined by qPCR. RESULTS: Our findings indicate increased infiltration of PDCs in breast cancer tissues of pLN patients than nLN patients, which correlates with CXCR4+ cells percentage. Interestingly, SDF-1 is highly immunostained in lymph nodes of pLN patients compared to nLN patients. Our in vitro experiments demonstrate an upregulation of NF-κB expression and CXCR4 cells upon stimulation with PDCs secretome of pLN patients than those of nLN patients. Also, PDCs isolated from pLN patients exhibited a higher TNF-α mRNA expression than nLN patients. Treatment of MCF-7 cell lines with TNF-α significantly upregulates CXCR4 expression. CONCLUSIONS: Our findings suggest a potential role for microenvironmental PDCs in breast cancer lymph node metastasis via CXCR4/SDF-1 axis.
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Neoplasias da Mama/patologia , Quimiocina CXCL12/metabolismo , Células Dendríticas/patologia , Metástase Linfática/patologia , NF-kappa B/metabolismo , Receptores CXCR4/metabolismo , Fator de Necrose Tumoral alfa/genética , Adulto , Idoso , Neoplasias da Mama/sangue , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Células Dendríticas/metabolismo , Feminino , Humanos , Metástase Linfática/genética , Células MCF-7 , Pessoa de Meia-Idade , Transdução de Sinais , Microambiente TumoralRESUMO
BACKGROUND: Persistence of hepatitis C virus (HCV) infection and response to antiviral therapy has been shown to be associated with inappropriate levels of cytokines and microRNAs (miRNAs). miRNA levels have been reported to fluctuate during treatment. Thus they could be useful predictors for responses to treatment among HCV infected patients, thereby reducing ineffective treatments. AIM: The current study aimed to investigate the relation between miRNA-21 expression profiles, transforming growth factor ß (TGF-ß) serum levels and response to treatment with the new direct antiviral drugs (sofosbuvir + daclatasvir ± ribavirin), among HCV infected Egyptian patients. SUBJECTS AND METHODS: This prospective study was conducted on 50 HCV infected patients (before and after treatment) and 20 healthy volunteers. miRNA expression profiles were determined by real-time polymerase chain reaction and TGF-ß1 serum levels were measured by using enzyme-linked immunosorbent assay. RESULTS: There was a significant increase in serum albumin, platelets count and a significant decrease in liver enzymes, serum bilirubin, and prothrombin time after treatment. Significant reduction of viral load among HCV patients after receiving the treatment was reported. Concomitantly, there was an increase in the relative quantity of miRNA-21 (P = .001*) and serum levels of TGF-ß1 ( P = .337) among HCV patients after receiving treatment. CONCLUSION: Nearly all responders to direct antiviral drugs showed increased levels of both miRNA-21 and TGF-ß1. This may indicate an interplay between TGF-ß1 and miRNA-21 during remission or progression of viral infection. Thus miRNA-21 could be used as promising serum biomarker, for assessment of antiviral treatment efficacy and improvement of fibrosis among chronically infected HCV patients.
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Antivirais/uso terapêutico , Hepatite C Crônica/tratamento farmacológico , MicroRNAs/sangue , Fator de Crescimento Transformador beta/sangue , Adulto , Biomarcadores/sangue , Carbamatos , Quimioterapia Combinada , Feminino , Hepacivirus/efeitos dos fármacos , Humanos , Imidazóis/uso terapêutico , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Pirrolidinas , Ribavirina/uso terapêutico , Sofosbuvir/uso terapêutico , Resultado do Tratamento , Valina/análogos & derivados , Carga Viral/efeitos dos fármacosRESUMO
Thirty-three Pseudomonas aeruginosa isolates, resistant to one or more ß-lactams, were included in this study. Identification of tested strains was confirmed using MALDI-TOF/MS. Phenotypic and genotypic ß-lactamase patterns were investigated. Most of the isolates were resistant to carbapenems (32 out of 33) and to the extended-spectrum cephalosporins (ESC) (30 out of 33). Phenotypically, the production of extended-spectrum beta-lactamase (ESBL), metallo-ß-lactamases (MBL), and carbapenemases was detected in 10, 23, and 9 isolates, respectively. However, AmpC hyperproduction was not phenotypically detected among all isolates. Genotypically, ESBL and MBL encoding genes were detected in 23 and 27 isolates, respectively. Altogether 27 strains were detected as blaVIM positive and 16 strains carried blaOXA-10 gene. To the best of our knowledge, this is the first report of P. aeruginosa clinical isolates harboring blaVEB together with blaGES in Egypt, where 5 of our 30 ESC-resistant isolates showed this genotype. Our results confirmed that resistance of P. aeruginosa isolates to ß-lactam antibiotics is mediated via multiple ß-lactamases belonging to different molecular classes. To the best of our knowledge, this is the first report of blaVEB among P. aeruginosa clinical isolates from Egypt. Ten isolates harbored blaVEB and five of them co-harbored blaVEB together with blaGES, blaVIM, and blaOXA-10.
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Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/enzimologia , Pseudomonas aeruginosa/isolamento & purificação , beta-Lactamases/análise , beta-Lactamases/genética , Técnicas Bacteriológicas , Egito , Genótipo , Humanos , Fenótipo , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/genética , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Resistência beta-Lactâmica , beta-Lactamases/classificaçãoRESUMO
PURPOSE: This report compares the risk factors, the tumor phenotypes, and the BRCA1/BRCA2 genotype of early onset breast cancer (EOBC) patients between Southern Europe and North Africa. METHODS: Four hundred and fifty six women with invasive EOBC (≤40 years) were prospectively included from four centers in France (n = 270) and four centers in North Africa (Algeria, Egypt, Morocco, Tunisia; n = 186). Life style, tumor phenotype, familial history, BRCA1/BRCA2 genotype were compared between the two populations. RESULTS: We found an older age at menarche, a higher number of childbearing, a more frequent breastfeeding, a higher body mass index, a lower use of oral contraceptives in North African women compared to French women. TNM stage at diagnosis was higher in North African women than in French women. North African women had a lower incidence of triple negative and proliferative (Ki 67 index > 20%) tumors. There was a lower rate of BRCA1 mutation in North Africa (7 vs. 15%, P = 0.02). Three putative BRCA1/2 founder mutations were identified in North Africa. CONCLUSIONS: In EOBC, we found significant differences in risk factors, phenotype and a higher incidence of BRCA1 mutations in Southern Europe as compared to North Africa. The worst prognosis previously reported for EOBC in North Africa is more likely due to a higher stage at diagnosis than to a more aggressive phenotype, since triple negative tumors are more common in Southern Europe and advanced tumors in North Africa.
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Proteína BRCA1/genética , Proteína BRCA2/genética , Neoplasias da Mama/patologia , África do Norte , Idade de Início , Neoplasias da Mama/genética , Feminino , França , Genótipo , Humanos , Estadiamento de Neoplasias , Estudos Prospectivos , Fatores de RiscoRESUMO
The current work presents a new structure based on Au/PVA/SiO2/p-Si/Al that has not been studied before. An aqueous solution of polyvinyl alcohol (PVA) polymer gel was deposited on the surface of SiO2/Si using the spin-coating technique. The silicon wafer was left to be oxidized in a furnace at 1170 k for thirty minutes, creating an interdiffusion layer of SiO2. The variations in the dielectric constant (Ð'), dielectric loss (Ðâ³), and dielectric tangent (tanδ) with the change in the frequency, voltage, and temperature were analyzed. The results showed an increase in the dielectric constant (Ð') and a decrease in the dielectric loss (Ðâ³) and tangent (tanδ); thus, the Au/PVA/SiO2/p-Si/Al heterostructure has opened up new frontiers for the semiconductor industry, especially for capacitor manufacturing. The Cole-Cole diagrams of the Ðâ³ and Ð' have been investigated at different temperatures and voltages. The ideality factor (n), barrier height (Φb), series resistance (Rs), shunt resistance (Rsh), and rectification ratio (RR) were also measured at different temperatures.
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BACKGROUND: The pathogenesis of inflammatory bowel disease (IBD) and colorectal cancer (CRC) is thought to be related to immune response against gut microbiota. TLR4, IgA, and EpCAM have a role in intestinal local immune response and their altered expression related to both IBD and CRC. Lipopolysaccharide (LPS) is the main activator of TLR4. The objective of this study is to evaluate the possible role of intestinal microbiota in the pathogenesis of IBD and CRC through expression of TLR4, IgA and EpCAM. METHODS: One hundred five cases were divided into (Group 1/ Control: 10 sections of normal colonic mucosa, Group 2/CRC: 51 cases, Group 3/IBD: 44 cases). Immunohistochemistry for TLR4, IgA, and EpCAM was done. LPS was assessed in all groups. TLR4 gene and protein expression were assessed in colorectal cancer cell line by RT-PCR and immunocytochemistry. RESULTS: There was a significant correlation between TLR4 and tumor grade (P value 0.003 and 0.01 respectively). A significant correlation was found between IgA expression and T stage (P value 0.02) and between EpCAM expression and histologic type (P value 0.02). In comparison of CRC patients to controls; there was a statistically significant different expression of TLR4 positivity, IgA positivity and EpCAM (P value <0.001, 0.004, <0.001 respectively). Patients with CRC were compared to colitis patients and there was a statistically significant different expression of IgA positivity and EpCAM expression (P value <0.001). There was significant higher expression of TLR4 in CRC cell line than the fibroblast by both PCR and immunocytochemistry (P-value: 0.003 and 0.024 respectively). LPS level in CRC patients was significantly higher than the control and IBD groups (P values <0.001 and <0.001 respectively). CONCLUSION: TLR4, IgA, EpCAM expression in both CRC and IBD might be related to the pathogenic role of microbiota and could represent potential prevention modalities and therapeutic targets.
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Neoplasias Colorretais , Doenças Inflamatórias Intestinais , Microbiota , Humanos , Neoplasias Colorretais/patologia , Receptor 4 Toll-Like/genética , Lipopolissacarídeos , Molécula de Adesão da Célula Epitelial/genética , Doenças Inflamatórias Intestinais/metabolismo , Imunoglobulina ARESUMO
BACKGROUND: Trastuzumab is an effective therapeutic approach for HER2-positive metastatic breast cancer (BC). However, a considerable number of patients develop resistance along the course of the disease. PTEN rs701848 polymorphisms are associated with an increased risk of developing cancer and have a potential role in predicting drug resistance. OBJECTIVE: We studied the significance of PTEN rs701848 variants as significant predictors for trastuzumab resistance in HER2-positive metastatic BC patients. Therefore, considering their value in predicting clinical outcomes. MATERIALS AND METHODS: This case-control study was conducted among female patients with HER2-positive metastatic breast cancer who underwent Trastuzumab therapy during the period from March 2017 to December 2020. PTEN rs701848 genotypes were analyzed in 160 HER2-positive metastatic breast cancer who received Trastuzumab therapy and clinically monitored for therapeutic response. RESULTS: PTEN rs701848 is deemed a significant predictor of Trastuzumab resistance and an independent prognostic factor of progression-free survival (PPFS). In particular, the C allele is associated with increased risk for Trastuzumab resistance and shorter PFS as compared to the homozygous TT genotype. CONCLUSION: PTEN rs701848 is significant predictor of trastuzumab resistance. Therefore, their value in predicting clinical outcomes is recommended.
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Neoplasias da Mama , Humanos , Feminino , Trastuzumab/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Intervalo Livre de Progressão , Estudos de Casos e Controles , Receptor ErbB-2/genética , Receptor ErbB-2/uso terapêutico , PTEN Fosfo-Hidrolase/genética , PTEN Fosfo-Hidrolase/uso terapêuticoRESUMO
Background & Aims: Chronic HCV infection causes cellular stress, fibrosis and predisposes to hepatocarcinogenesis. Mitochondria play key roles in orchestrating stress responses by regulating bioenergetics, inflammation and apoptosis. To better understand the role of mitochondria in the viral life cycle and disease progression of chronic hepatitis C, we studied morphological and functional mitochondrial alterations induced by HCV using productively infected hepatoma cells and patient livers. Methods: Biochemical and imaging assays were used to assess localization of cellular and viral proteins and mitochondrial functions in cell cultures and liver biopsies. Cyclophilin D (CypD) knockout was performed using CRISPR/Cas9 technology. Viral replication was quantified by quantitative reverse-transcription PCR and western blotting. Results: Several HCV proteins were found to associate with mitochondria-associated endoplasmic reticulum (ER) membranes (MAMs), the points of contact between the ER and mitochondria. Downregulation of CypD, which is known to disrupt MAM integrity, reduced viral replication, suggesting that MAMs play an important role in the viral life cycle. This process was rescued by ectopic CypD expression. Furthermore, HCV proteins were found to associate with voltage dependent anion channel 1 (VDAC1) at MAMs and to reduce VDAC1 protein levels at MAMs in vitro and in patient biopsies. This association did not affect MAM-associated functions in glucose homeostasis and Ca2+ signaling. Conclusions: HCV proteins associate specifically with MAMs and MAMs play an important role in viral replication. The association between viral proteins and MAMs did not impact Ca2+ signaling between the ER and mitochondria or glucose homeostasis. Whether additional functions of MAMs and/or VDAC are impacted by HCV and contribute to the associated pathology remains to be assessed. Impact and implications: Hepatitis C virus infects the liver, where it causes inflammation, cell damage and increases the long-term risk of liver cancer. We show that several HCV proteins interact with mitochondria in liver cells and alter the composition of mitochondrial subdomains. Importantly, HCV requires the architecture of these mitochondrial subdomains to remain intact for efficient viral replication.
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Viruses play an important role in cancer development as about 12% of cancer types are linked to viral infections. Viruses that induce cellular transformation are known as oncoviruses. Although the mechanisms of viral oncogenesis differ between viruses, all oncogenic viruses share the ability to establish persistent chronic infections with no obvious symptoms for years. During these prolonged infections, oncogenic viruses manipulate cell signaling pathways that control cell cycle progression, apoptosis, inflammation, and metabolism. Importantly, it seems that most oncoviruses depend on these changes for their persistence and amplification. Metabolic changes induced by oncoviruses share many common features with cancer metabolism. Indeed, viruses, like proliferating cancer cells, require increased biosynthetic precursors for virion production, need to balance cellular redox homeostasis, and need to ensure host cell survival in a given tissue microenvironment. Thus, like for cancer cells, viral replication and persistence of infected cells frequently depend on metabolic changes. Here, we draw parallels between metabolic changes observed in cancers or induced by oncoviruses, with a focus on pathways involved in the regulation of glucose, lipid, and amino acids. We describe whether and how oncoviruses depend on metabolic changes, with the perspective of targeting them for antiviral and onco-therapeutic approaches in the context of viral infections.
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Introduction. Tuberculosis (TB) is a great public health problem in developing countries such as Egypt. Genotyping of Mycobacterium tuberculosis isolates has a prominent role in the field of TB prevention.Aim. This study aimed to evaluate real-time PCR using Minor Groove Binder (MGB) probes and to identify circulating lineages/sub-lineages of M. tuberculosis and their transmission patterns.Hypothesis. We hypothesize that MIRU-VNTR technique is efficient in identifying circulating M. tuberculosis lineages in Egypt.Methodology. Fifty sputum specimens positive for acid-fast bacilli were included. Isoniazid (INH) resistance was detected using the 1â% proportion method. Real-time PCR using MGB-probes was used for simultaneous detection of TB infection and INH resistance. Partial sequencing of the katG gene was used to confirm INH resistance results. A standard 15 Mycobacterial Interspersed Repetitive Unit Variable Number Tandem Repeat (15-MIRU-VNTR) approach was used for genotyping through the MIRU-VNTRplus online platform.Results. Only seven specimens showed phenotypic resistance to INH. M. tuberculosis was detected in all samples, while a mutation in the katG gene codon 315 was detected only in five samples, which were also phenotypically INH-resistant. Sequencing of the katG gene showed codon 315 mutation genotypically and phenotypically in the five INH-resistant isolates. Molecular genotyping of M. tuberculosis isolates revealed that the majority of isolates (26/50, 52â%) belonged to the S family of lineage_4. A low clustering rate (2â%) was observed among our isolates. According to the Hunter-Gaston Discriminatory Index (HGDI), 11 MIRU-VNTR loci were highly or moderately discriminative, while four loci were less polymorphic.Conclusion. MIRU-VNTR genotyping revealed a low clustering rate with a low recent transmission rate of M. tuberculosis strains in Alexandria, Egypt.
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Mycobacterium tuberculosis , Tuberculose , Humanos , Técnicas de Tipagem Bacteriana/métodos , DNA Bacteriano/genética , Genótipo , Isoniazida/farmacologia , Repetições Minissatélites , Mycobacterium tuberculosis/genética , Reação em Cadeia da Polimerase em Tempo Real , Tuberculose/microbiologiaRESUMO
BACKGROUND: Human immunodeficiency virus type 1 (HIV-1) is characterized by high genetic diversity due to its high mutation and recombination rates. Although, there is an increasing prevalence of Circulating Recombinant Forms (CRFs) worldwide, subtype B is still recognized as the predominant subtype in the Middle East and North Africa (MENA) region. There is a limited sampling of HIV in this region due to its low prevalence. The main purpose of this study is to provide a summary of the current status of the resident HIV subtypes and their distribution among Egyptian patients. METHODOLOGY: Forty-five HIV-1 patients were included in this study. Partial pol gene covering the protease (PR) and Reverse Transcriptase (RT) was successfully amplified in 21 HIV patients using nested PCR of cDNA of the viral genomic RNA, then sequenced. The sequence data were used for viral HIV-1 subtyping by 5 online subtyping tools: NCBI viral genotyping tool, Stanford University HIV database (HIVDB) subtyping program, REGA tool, Context-Based Modeling for Expeditious Typing (COMET) tool, and Recombinant Identification Program (RIP) tool. The final subtype assignment was based on molecular phylogenetic analysis. RESULTS: Unexpectedly, non-B subtypes are dominating, with the most common circulating one is CRF02_AG (57.1%) followed by subtype B (14.3%), subtype BG recombinant (9.5%), CRF35_ AD (9.5%), subtype A1 and CRF06_cpx (4.8% each). CONCLUSION: To the best of our knowledge, this is the first study to tackle HIV-1 subtyping among the group of HIV-1 patients in Egypt. CRF02_AG is the most prevalent subtype in Egypt.
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Infecções por HIV , HIV-1 , Egito/epidemiologia , Infecções por HIV/epidemiologia , HIV-1/genética , Humanos , Epidemiologia Molecular , Filogenia , RNA Viral/genéticaRESUMO
Hepatocellular carcinoma (HCC) is a prevalent liver cancer representing the fourth most lethal cancer worldwide. Trans-Resveratrol (T-R) possesses a promising anticancer activity against HCC. However, it suffers from poor bioavailability because of the low solubility, chemical instability, and hepatic metabolism. Herein, we developed T-R-loaded nanocochleates using a simple trapping method. Nanocarriers were optimized using a comprehensive in-vitro characterization toolset and evaluated for the anticancer activity against HepG2 cell line. T-R-loaded nanocochleates demonstrated monodispersed cylinders (163.27 ± 2.68 nm and 0.25 ± 0.011 PDI) and -46.6 mV ζ-potential. They exhibited a controlled biphasic pattern with minimal burst followed by sustained release for 72 h. Significant enhancements of Caco-2 transport and ex-vivo intestinal permeation over liposomes, with 1.8 and 2.1-folds respectively, were observed. Nanocochleates showed significant reduction of 24 h IC50 values compared to liposomes and free T-R. Moreover, an efficient knockdown of anti-apoptotic (Bcl-2) and cancer stemness (NANOG) genes was demonstrated. To the best of our knowledge, we are the first to develop T-R loaded nanocochleates and scrutinize its potential in suppressing NANOG expression, 2-folds lower, compared to free T-R. According to these auspicious outcomes, nanocochleates represent a promising nanoplatform to enhance T-R oral permeability and augment its anticancer efficacy in the treatment of HCC.
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Carcinoma Hepatocelular/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Nanopartículas , Resveratrol/administração & dosagem , Administração Oral , Animais , Antineoplásicos Fitogênicos/administração & dosagem , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/farmacologia , Células CACO-2 , Células Hep G2 , Humanos , Lipossomos , Masculino , Tamanho da Partícula , Ratos , Ratos Wistar , Resveratrol/química , Resveratrol/farmacologia , SolubilidadeRESUMO
INTRODUCTION: Tumor microenvironment plays crucial role in cancer evolution. There is a dynamic and continuous relation between immune cells and cancer cells' resistance. Tumor infiltration CD8-lymphocytes and programmed death ligand-1 have proved important prognostic role in different malignancies. We aimed at evaluating this role in laryngeal cancer. PATIENTS AND METHODS: We prospectively analyzed laryngeal cancer patients' specimens, to identify the CD8-lymphocytes and the PD-L1 expression. A total score formed of the sum of percentage and intensity of PD-L1. A final rate was considered as negative or low when combined percentage and intensity scores 0 to 4, and high when scores 5-7. CD8-lymphocyte infiltration was divided into strong (= 10/100 of epithelial cells or =20/100 stromal cell infiltration) or weak (<10/100 epithelial cells or <20/100 stromal cell infiltration). RESULTS: Forty patients were included; twelve had stage 1 or 2 and 28 with advanced stages. PD-L1 expressionwas positive in 92.5%. Neither the PD-L1 nor CD8- lymphocytes had overall survival impact, however high PD-L1 correlated with better survival in advanced stage subgroup (p = 0.036), high CD8-lymphocytes infiltration had better survival but did not reach significance. Therewas significant correlation between the CD8-lymphocyte infiltration; whether epithelial or stromal, and tumor PD-L1 expression; p-value of 0.001 and < 0.0001 respectively. Subgroup of patients with low CD8+ infiltration and low PD-L1 had the worst survival. CONCLUSION: There is a correlation between CD8- l lymphocytes infiltration and PD-L1 expression inlaryngeal cancer and high PD-L1 expression is associated with better OS in advanced stages. Key words: PD-L1, CD8, laryngeal cancer, tumor microenvironment.
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Antígeno B7-H1 , Neoplasias Laríngeas , Linfócitos T CD8-Positivos , Humanos , Linfócitos do Interstício Tumoral , Prognóstico , Estudos Prospectivos , Microambiente TumoralRESUMO
BACKGROUND: Besides its antimicrobial action, doxycycline (DX) has lately been repurposed as a small-molecule drug for osteogenic purposes. However, osteogenic DX application is impeded by its dose-dependent cytotoxicity. Further, high-dose DX impairs cell differentiation and mineralization. PURPOSE: Integrating DX into a biomaterial-based delivery system that can control its release would not only ameliorate its cytotoxic actions but also augment its osteogenic activity. In this work, we managed to engineer novel composite DX-hydroxyapatite-polycaprolactone nanoparticles (DX/HAp/PCL) to modify DX osteogenic potential. METHODS: Employing a 23-factorial design, we first optimized HApN for surface-area attributes to maximize DX loading. Composite DX/HAp/PCL were then realized using a simple emulsification technique, characterized using various in vitro methods, and evaluated for in vitro osteogenesis. RESULTS: The developed HApN exhibited a favorable crystalline structure, Ca:P elemental ratio (1.67), mesoporous nature, and large surface area. DX/HAp/PCL achieved the highest reported entrapment efficiency (94.77%±1.23%) of DX in PCL-based particles. The developed composite system achieved controlled release of the water-soluble DX over 24 days. Moreover, the novel composite nanosystem managed to significantly ameliorate DX cytotoxicity on bone-marrow stem cells, as well as enhance its overall proliferation potential. Alkaline phosphatase and mineralization assays revealed superior osteodifferentiation potential of the composite system. Quantification of gene expression demonstrated that while DX solution was able to drive bone-marrow stem cells down the osteogenic lineage into immature osteoblasts after 10-day culture, the innovative composite system allowed maturation of osteodifferentiated cells. To the best of our knowledge, this is the first work to elaborate the impact of DX on the expression of osteogenic genes: RUNX2, OSP, and BSP. Further, the osteogenicity of a DX-loaded particulate-delivery system has not been previously investigated. CONCLUSION: Our findings indicate that repurposing low-dose DX in complementary biomaterial-based nanosystems can offer a prominent osteogenic candidate for bone-regeneration purposes.