Cryopreserved human hepatocytes were used to investigate the role of arylamine-acetyltransferase 2 (NAT2; EC 2.3.1.5) polymorphism on the-acetylation of isoniazid (INH).genotype was determined by Taqman allelic discrimination assay and INH-acetylation was measured by high performance liquid chromatography. INH-acetylation ratesexhibited a robust and highly significant (<0.005) NAT2 phenotype-dependent metabolism.-acetylation rateswere INH concentration- and time-dependent. Following incubation for 24 h with 12.5 or 100 µmol/L INH, acetyl-INH concentrations varied significantly (= 0.0023 and= 0.0002) across cryopreserved human hepatocytes samples from rapid, intermediate, and slow acetylators, respectively. The clear association betweengenotype and phenotype supports use ofgenotype to guide INH dosing strategies in the treatment and prevention of tuberculosis.

BACKGROUND: Bipolar hip hemiarthroplasty is used in the management of fractures of the proximal femur. The dual articulation is cited as advantageous in comparison to unipolar prostheses as it decreases acetabular erosion, has a lower dislocation rates and is easier to convert to a total hip arthroplasty (THA) should the need arise. However, these claims are debatable. Our study examines the rate of conversion of the bipolar hemiarthroplasty to THA and the justification for using it on the basis of future conversion to THA. METHODS: All cases of bipolar hemiarthroplasty performed in our unit for hip fractures over a 9-year period (1999-2007) were reviewed. Medical notes and radiographs of all patients were reviewed, and all surviving patients that were contactable received a telephone follow-up. RESULTS: Of all 164 patients reviewed with a minimum of 1 year from date of surgery, 4 patients had undergone a conversion of their bipolar prosthesis to THA. Three conversions were performed for infection, dislocation, and fracture. Only one (0.6%) conversion was performed for groin pain. CONCLUSIONS: Our study show that bipolar hemiarthroplasties for hip fractures have a low conversion rate to THAs and this is comparable to the published conversion rate of unipolar hemiarthroplasties.
Aged ; Arthroplasty, Replacement, Hip/*methods ; Female ; Hemiarthroplasty/*methods ; Hip Fractures/*surgery ; Humans ; Male ; Reoperation ; Retrospective Studies ; Treatment Outcome

PURPOSE: Based in a hospital serving one of the most deprived areas in the United Kingdom (UK), we aimed to investigate, using the Indices of Deprivation 2010, the hypothesis that deprivation affects the stage and mode of presentation of colorectal cancer. METHODS: All newly diagnosed patients with colorectal cancer presenting to a District General Hospital in the UK between January 2010 and December 2014 were included. Data were collected from the Somerset National Cancer Database. The effect of social deprivation, measured using the Index of Multiple Deprivation Score, on the stage and mode of presentation was evaluated utilizing Microsoft Excel and IBM SPSS ver. 22.0. RESULTS: A total of 701 patients (54.5% male; mean age, 76 years) were included; 534 (76.2%) underwent a surgical procedure, and 497 (70.9%) underwent a colorectal resection. Of the patients undergoing a colorectal resection, 86 (17.3%) had an emergency surgical resection. Social deprivation was associated with Duke staging (P = 0.09). The 90-day mortality in patients undergoing emergency surgery was 12.8% compared to 6.8% in patients undergoing elective surgery (P = 0.06). No association was found between deprivation and emergency presentation (P = 0.97). A logistic regression analysis showed no increase in the probability of metastasis amongst deprived patients. CONCLUSION: This study suggests an association between deprivation and the stage of presentation of colorectal cancer. Patients undergoing emergency surgery tend to have a higher 90-day mortality rate, although this was not related to deprivation. This study highlights the need to develop an individual measure to assess social deprivation.
Colorectal Neoplasms* ; Emergencies ; Great Britain ; Hospitals, General ; Humans ; Logistic Models ; Male ; Mortality ; Neoplasm Metastasis ; Social Class

Purpose: Accurate component placement and restoration of patient anatomy are critical in total hip arthroplasty (THA) surgery. Although intraoperative radiographs are sometimes utilized, it is unclear whether this practice can improve accuracy. Materials and Methods: This study evaluated acetabular cup abduction, anteversion, leg length, and offset among 100 posterior approach THAs performed without imaging (No X-ray group) and compared them to a subsequent series of 100 THAs where an intraoperative radiograph was taken with the trial components in place (X-ray group). THAs were performed using a posterior approach by a single, experienced surgeon whose goal was to place the cup at 45° of abduction and 30° of anteversion. Supine anteroposterior pelvic digital radiographs taken at the first (nominal 4-week) postoperative visit were used for measurements. Results: Slight differences in cup abduction (47°±6° vs 44°±6°, respectively, P=0.003) and anteversion angle (35°±6° vs 31°±6°, respectively, P<0.001) were observed between the X-ray and No X-ray groups; however, a similar proportion of cups within 10° of the target angles was observed (76% vs 83%, respectively, P=0.22). No difference in offset measurements (1.1±6.6 mm vs 0.3±6.9 mm, respectively, P=0.42) or leg lengths (0.3±3.8 mm vs 0.3±4.8 mm, respectively, P=0.94) was observed between the X-ray and No X-ray groups; however, the X-ray group showed less leg length variation (P=0.05). Conclusion In this study, the routine use of intraoperative radiographs was not associated with improved implant positioning for uncomplicated primary THA.

E2F-1 and p53 are sequence specific transcription factors that are intimately involved in the regulation of the cell cycle. In addition to their role in cell cycle control, both E2F-1 and p53 have been identified as tumor suppressors and mediators of apoptosis. We have shown previously that adenoviral-mediated E2F-1 overexpression induces efficient apoptosis in colon adenocarcinoma cells. Previous reports have suggested that E2F-1 and p53 cooperate to mediate apoptosis and therefore, in this study, we examined the efficacy of combination gene therapy using adenovirus vectors expressing E2F-1 and p53 in human colon adenocarcinoma cell lines, HT-29 and SW620 (both mutant p53). Cells were treated by mock infection or infection with adenoviral vectors expressing b-galactosidase (LacZ), E2F-1, p53 or a combination of E2F-1 and p53. IC25 concentrations of each virus were estimated and used for each treatment in order to detect any synergistic or cooperative effects on tumor cell death in the combination therapy. By 5 days post infection, E2F-1-overexpressing cells exhibited growth inhibition and approximately 40-50% cell death in both cell lines. Co-expression of p53 with E2F-1 abrogated E2F-1-mediated growth inhibition and cell death. Cell cycle analysis revealed that overexpression of E2F-1 resulted in an accumulation of cells in G2/M phase, while overexpression of p53 resulted in a G1 phase accumulation. However, co-expression of E2F-1 and p53 counteracted each other as fewer cells accumulated in G1 and G2/M when compared to either p53 or E2F-1 alone. Furthermore, co-expression of p53 with E2F-1 resulted in decreased levels of E2F-1 protein expression. Mechanistically, upregulation of the CDK inhibitory protein, p21(WAF1/CIP1), was demonstrated in HT-29 cells following overexpression of either E2F-1, p53 or the combination E2F-1/p53 therapy. However, in SW620 cells, only the cells infected with Ad-p53 alone or in combination resulted in upregulation of p21(WAF1/CIP1). These results suggest that p53 and p21(WAF1/CIP1) may cooperate to inhibit the expression and activity of E2F-1. In conclusion, combination adenoviral vector-mediated E2F-1 and p53 gene transfer was not therapeutically advantageous in this in vitro model of human colon adenocarcinoma.
Adenocarcinoma/*metabolism/pathology ; Adenoviridae/genetics ; Apoptosis/*physiology ; Cell Cycle ; Cell Division ; Colonic Neoplasms/*metabolism/pathology ; Comparative Study ; Cyclins ; Gene Expression ; Gene Therapy ; Gene Transfer Techniques ; *Genes, p53 ; Genetic Vectors ; HT29 Cells ; Human ; Protein p53/genetics/*metabolism ; Recombinant Proteins/metabolism ; Transcription Factors/genetics/metabolism/physiology ; Tumor Cells, Cultured ; Up-Regulation

E2F-1 and p53 are sequence specific transcription factors that are intimately involved in the regulation of the cell cycle. In addition to their role in cell cycle control, both E2F-1 and p53 have been identified as tumor suppressors and mediators of apoptosis. We have shown previously that adenoviral-mediated E2F-1 overexpression induces efficient apoptosis in colon adenocarcinoma cells. Previous reports have suggested that E2F-1 and p53 cooperate to mediate apoptosis and therefore, in this study, we examined the efficacy of combination gene therapy using adenovirus vectors expressing E2F-1 and p53 in human colon adenocarcinoma cell lines, HT-29 and SW620 (both mutant p53). Cells were treated by mock infection or infection with adenoviral vectors expressing b-galactosidase (LacZ), E2F-1, p53 or a combination of E2F-1 and p53. IC25 concentrations of each virus were estimated and used for each treatment in order to detect any synergistic or cooperative effects on tumor cell death in the combination therapy. By 5 days post infection, E2F-1-overexpressing cells exhibited growth inhibition and approximately 40-50% cell death in both cell lines. Co-expression of p53 with E2F-1 abrogated E2F-1-mediated growth inhibition and cell death. Cell cycle analysis revealed that overexpression of E2F-1 resulted in an accumulation of cells in G2/M phase, while overexpression of p53 resulted in a G1 phase accumulation. However, co-expression of E2F-1 and p53 counteracted each other as fewer cells accumulated in G1 and G2/M when compared to either p53 or E2F-1 alone. Furthermore, co-expression of p53 with E2F-1 resulted in decreased levels of E2F-1 protein expression. Mechanistically, upregulation of the CDK inhibitory protein, p21(WAF1/CIP1), was demonstrated in HT-29 cells following overexpression of either E2F-1, p53 or the combination E2F-1/p53 therapy. However, in SW620 cells, only the cells infected with Ad-p53 alone or in combination resulted in upregulation of p21(WAF1/CIP1). These results suggest that p53 and p21(WAF1/CIP1) may cooperate to inhibit the expression and activity of E2F-1. In conclusion, combination adenoviral vector-mediated E2F-1 and p53 gene transfer was not therapeutically advantageous in this in vitro model of human colon adenocarcinoma.
Adenocarcinoma/*metabolism/pathology ; Adenoviridae/genetics ; Apoptosis/*physiology ; Cell Cycle ; Cell Division ; Colonic Neoplasms/*metabolism/pathology ; Comparative Study ; Cyclins ; Gene Expression ; Gene Therapy ; Gene Transfer Techniques ; *Genes, p53 ; Genetic Vectors ; HT29 Cells ; Human ; Protein p53/genetics/*metabolism ; Recombinant Proteins/metabolism ; Transcription Factors/genetics/metabolism/physiology ; Tumor Cells, Cultured ; Up-Regulation

Mannan-binding lectin (MBL) is a soluble innate immune protein that binds to glycosylated targets. MBL acts as an opsonin and activates complement, contributing to the destruction and clearance of infecting microorganisms. Hepatitis C virus (HCV) encodes two envelope glycoproteins E1 and E2, expressed as non-covalent E1/E2 heterodimers in the viral envelope. E1 and E2 are potential ligands for MBL. Here we describe an analysis of the interaction between HCV and MBL using recombinant soluble E2 ectodomain fragment, the full-length E1/E2 heterodimer, expressed in vitro, and assess the effect of this interaction on virus entry. A binding assay using antibody capture of full length E1/E2 heterodimers was used to demonstrate calcium dependent, saturating binding of MBL to HCV glycoproteins. Competition with various saccharides further confirmed that the interaction was via the lectin domain of MBL. MBL binds to E1/E2 representing a broad range of virus genotypes. MBL was shown to neutralize the entry into Huh-7 cells of HCV pseudoparticles (HCVpp) bearing E1/E2 from a wide range of genotypes. HCVpp were neutralized to varying degrees. MBL was also shown to neutralize an authentic cell culture infectious virus, strain JFH-1 (HCVcc). Furthermore, binding of MBL to E1/E2 was able to activate the complement system via MBL-associated serine protease 2. In conclusion, MBL interacts directly with HCV glycoproteins, which are present on the surface of the virion, resulting in neutralization of HCV particles.
Binding, Competitive ; Glycosylation ; Hepacivirus ; genetics ; pathogenicity ; physiology ; Humans ; Mannose-Binding Lectin ; metabolism ; Mannose-Binding Protein-Associated Serine Proteases ; metabolism ; Monosaccharides ; metabolism ; Protein Binding ; Protein Multimerization ; Tumor Cells, Cultured ; Viral Envelope Proteins ; metabolism ; Virion ; pathogenicity ; physiology ; Virus Internalization