Genetic and metabolic studies towards personalized conditioning regimen prior to stem cell transplantation

Hematopoietic stem cell transplantation (HSCT) is a curative treatment for several malignant and non- malignant diseases. The busulphan (Bu)/cyclophosphamide (Cy) combination is one of the most common conditioning regimens given prior to HSCT. The general aim of the present thesis is to investigate the molecular mechanisms underlying the metabolism of the Bu/Cy conditioning regimen in order to personalize the treatment and improve the clinical outcome. To follow the metabolic pathway of busulphan, a new gas chromatography-mass spectrometry (GC-MS) method was developed and validated for the quantification and detection of busulphan and its four major metabolites. Incubation of the first core metabolite of busulphan, tetrahydrothiophene (THT), with human liver microsomes or recombinant enzymes has resulted in the formation of subsequent metabolites. The highest initial THT disappearance rate and the highest CLint value were observed with FMO3 followed by several CYPs indicating that FMO3 and, to a lesser extent, CYPs are involved in the metabolic pathway of busulphan. Moreover, FMO3 inhibition significantly (P < 0.05) affected Bu and THT kinetics in mice. In patients, FMO3 expression was significantly (P < 0.05) up-regulated during Bu treatment. In order to personalize oral Bu dosage, a reliable limited sampling model was developed and evaluated in both adult and pediatric patients. To understand the role of cyclophosphamide in the conditioning regimen, the gene expression profile over two days of Cy treatment was investigated, where 299 genes were found to be specifically affected by the treatment. Cyclophosphamide down-regulated the expression of several genes mapped to immune/autoimmune activation and graft rejection including CD3, CD28, CTLA4 and IL2R, and up-regulated immune-related receptor genes, e.g. IL1R2, IL18R1, and FLT3. Significant (P < 0.01) up-regulation, with high inter-individual variation, of the cytochrome P450 oxidoreductase (POR) gene was also observed during Cy treatment. In vitro, different batches of CYP2B6.1, with different ratios of POR/CYP, showed positive correlation between the intrinsic clearance (Vmax/Km) and the POR/CYP ratio for the Cy 4-hydroxylation. Further analysis of the above mentioned patients, prior to Cy treatment, revealed that CYP2J2 mRNA expression was significantly (P < 0.01) higher compared to healthy controls. CYP2J2 expression was further up-regulated during Cy treatment, with high inter-individual variation. Repeated treatment with Cy resulted in an increased 4-OH-Cy/Cy ratio, indicating auto-induction of Cy-metabolism. The viability of HL-60 cells, lacking CYP2B6 but expressing CYP2J2, was reduced after incubation with Cy. Inhibition of CYP2J2 reduced 4-OH-Cy formation and improved HL-60 cell survival. Cy incubation with recombinant CYP2J2 confirmed that CYP2J2 is involved in Cy bioactivation. In summary, the present results have improved our understanding of the Bu/Cy metabolism. This knowledge may help to interpret several interactions, high inter-individual variability, adverse effects and unexpected toxicity observed during and/or after the conditioning regimen. This certainly will help in developing new strategies for personalized medicine and thus improve clinical outcome

Optimization of Residential Load Consumption during Energy Peaks using Smart Metering

Abstract. Recently smart demand side management (DSM) is a very important tool that permits customers to take right decisions for their energy consumption and helps the energy utilities to decrease the over load demand and reshape the load curve. This paper proposes an optimized DSM technique based on smart metering uses different techniques such as load shifting and peak clipping to minimize domestic power consumption especially during load peaks. A new optimization technique (Bat Algorithm) is applied on proposed system and then compares results with other optimization techniques (Genetic Algorithm and Interior point Algorithm) to optimize the minimum consumption during peak hours according to load type. A control algorithm is applied to the proposed system to achieve the load shifting and load clipping according to the optimization results

Cognitive dysfunction among inpatients and outpatients with schizophrenia : relationship to positive and negative symptoms

Background: Cognitive impairment is an established feature of schizophrenia and is a strong predictor of eventual social and functional outcome. Few studies have investigated cognitive impairment in hospital long-stay patients with schizophrenia. This study evaluates and compares cognitive function among a sample of patients with schizophrenia in both inpatient and outpatient departments in order to determine the relationship between cognitive impairment and clinical variables. A cross-sectional comparative study based on a semi-structured interview investigating 100 inpatients with schizophrenia recruited from El-Abassia Mental Health Hospital departments compared to 100 patients with schizophrenia selected from the outpatients’ clinic matched with cases. The assessment tools included SCID-I, the Adult Wechsler Intelligence Scale, the computerized version of Wisconsin Card Sorting Test (WCST), Mini-Mental State Examination (MMSE), and Positive and Negative Syndrome Scale (PANSS). Results: Patients with schizophrenia showed significant deficits on cognitive function with no statistically significant difference between the inpatient and outpatient groups. Executive function was significantly correlated with verbal, non-verbal, and total IQ. Executive function was negatively correlated with the positive and general symptoms of PANSS and not correlated with its negative symptoms. In addition, we did not find any statistically significant relationship between cognitive functions and the duration of illness. Conclusion: The study provides evidence that institutionalization is not an influential factor on cognitive impairment patients with schizophrenia. However, the psychopathological aspects of the disorder are one of the crucial factors affecting the cognitive function in schizophrenia

Obligations of Contracts: Intent and Distortion

<p><b>Introduction:</b> Treosulfan is an alkylating agent that is used for the treatment of ovarian cancer and for conditioning prior to stem cell transplantation. It is a prodrug that is activated non-enzymatically to two active epoxides.</p> <p><b>Objectives:</b> To optimize a protocol for both <i>in vivo</i> samples handling and <i>in vitro</i> drug preparation. Treosulfan stability was tested in biological fluids at different conditions as well as for its cytotoxicity on cell lines.</p> <p><b>Results:</b> Plasma samples can be safely frozen for a short period up to 8 h, however; for longer periods, samples should be acidified. Urine samples and cell culture media can be safely frozen regardless their pH. For <i>in vitro</i> investigations, incubation of treosulfan at 37 °C for 24 h activated 100% of the drug. Whole blood acidification should be avoided for the risk of hemolysis. Finally; treosulfan cytotoxicity on HL-60 cells has increased following pre-incubation for 24 h at 37 °C compared to K562 cell line.</p> <p><b>Conclusion:</b> The stability profiling of treosulfan provided a valuable reference for handling of biological samples for both <i>in vivo</i> and <i>in vitro</i> studies. These results can be utilized for further investigations concerning the drug kinetics and dynamics in addition to the development of new pharmaceutical formulations.</p

A prospective dual-centre intra-individual controlled study for the treatment of burns comparing dermis graft with split-thickness skin auto-graft

Publisher Copyright: © 2022, The Author(s).To investigate if donor and recipient site morbidity (healing time and cosmesis) could be reduced by a novel, modified split-thickness skin grafting (STSG) technique using a dermal component in the STSG procedure (DG). The STSG technique has been used for 150 years in surgery with limited improvements. Its drawbacks are well known and relate to donor site morbidity and recipient site cosmetic shortcomings (especially mesh patterns, wound contracture, and scarring). The Dermal graft technique (DG) has emerged as an interesting alternative, which reduces donor site morbidity, increases graft yield, and has the potential to avoid the mesh procedure in the STSG procedure due to its elastic properties. A prospective, dual-centre, intra-individual controlled comparison study. Twenty-one patients received both an unmeshed dermis graft and a regular 1:1.5 meshed STSG. Aesthetic and scar assessments were done using The Patient and Observer Scar Assessment Scale (POSAS) and a Cutometer Dual MPA 580 on both donor and recipient sites. These were also examined histologically for remodelling and scar formation. Dermal graft donor sites and the STSG donor sites healed in 8 and 14 days, respectively (p < 0.005). Patient-reported POSAS showed better values for colour for all three measurements, i.e., 3, 6, and 12 months, and the observers rated both vascularity and pigmentation better on these occasions (p < 0.01). At the recipient site, (n = 21) the mesh patterns were avoided as the DG covered the donor site due to its elastic properties and rendered the meshing procedure unnecessary. Scar formation was seen at the dermal donor and recipient sites after 6 months as in the standard scar healing process. The dermis graft technique, besides potentially rendering a larger graft yield, reduced donor site morbidity, as it healed faster than the standard STSG. Due to its elastic properties, the DG procedure eliminated the meshing requirement (when compared to a 1:1.5 meshed STSG). This promising outcome presented for the DG technique needs to be further explored, especially regarding the elasticity of the dermal graft and its ability to reduce mesh patterns. Trial registration: ClinicalTrials.gov Identifier (NCT05189743) 12/01/2022.Peer reviewe

Cyclin A and cyclin D1 as significant prognostic markers in colorectal cancer patients

BACKGROUND: Colorectal cancer is a common cancer all over the world. Aberrations in the cell cycle checkpoints have been shown to be of prognostic significance in colorectal cancer. METHODS: The expression of cyclin D1, cyclin A, histone H3 and Ki-67 was examined in 60 colorectal cancer cases for co-regulation and impact on overall survival using immunohistochemistry, southern blot and in situ hybridization techniques. Immunoreactivity was evaluated semi quantitatively by determining the staining index of the studied proteins. RESULTS: There was a significant correlation between cyclin D1 gene amplification and protein overexpression (concordance = 63.6%) and between Ki-67 and the other studied proteins. The staining index for Ki-67, cyclin A and D1 was higher in large, poorly differentiated tumors. The staining index of cyclin D1 was significantly higher in cases with deeply invasive tumors and nodal metastasis. Overexpression of cyclin A and D1 and amplification of cyclin D1 were associated with reduced overall survival. Multivariate analysis shows that cyclin D1 and A are two independent prognostic factors in colorectal cancer patients. CONCLUSIONS: Loss of cell cycle checkpoints control is common in colorectal cancer. Cyclin A and D1 are superior independent indicators of poor prognosis in colorectal cancer patients. Therefore, they may help in predicting the clinical outcome of those patients on an individual basis and could be considered important therapeutic targets

Differentiation-associated urothelial cytochrome P450 oxidoreductase predicates the xenobiotic-metabolising activity of “luminal” muscle-invasive bladder cancers

Extra-hepatic metabolism of xenobiotics by epithelial tissues has evolved as a self-defence mechanism but has potential to contribute to the local activation of carcinogens. Bladder epithelium (urothelium) is bathed in excreted urinary toxicants and pro-carcinogens. This study reveals how differentiation affects cytochrome P450 (CYP) activity and the role of NADPH:P450 oxidoreductase (POR). CYP1A1 and CYP1B1 transcripts were inducible in normal human urothelial (NHU) cells maintained in both undifferentiated and functional barrier-forming differentiated states in vitro. However, ethoxyresorufin O-deethylation (EROD) activity, the generation of reactive BaP metabolites and BaP-DNA adducts, were predominantly detected in differentiated NHU cell cultures. This gain-of-function was attributable to the expression of POR, an essential electron donor for all CYPs, which was significantly upregulated as part of urothelial differentiation. Immunohistology of muscle-invasive bladder cancer (MIBC) revealed significant overall suppression of POR expression. Stratification of MIBC biopsies into luminal and basal groups, based on GATA3 and cytokeratin 5/6 labeling, showed POR over-expression by a subgroup of the differentiated luminal tumors. In bladder cancer cell lines, CYP1-activity was undetectable/low in basal PORlo T24 and SCaBER cells and higher in the luminal POR over-expressing RT4 and RT112 cells than in differentiated NHU cells, indicating that CYP-function is related to differentiation status in bladder cancers. This study establishes POR as a predictive biomarker of metabolic potential. This has implications in bladder carcinogenesis for the hepatic versus local activation of carcinogens and as a functional predictor of the potential for MIBC to respond to prodrug therapies

High Expression of Testes-Specific Protease 50 Is Associated with Poor Prognosis in Colorectal Carcinoma

Testes-specific protease 50 (TSP50) is normally expressed in testes and abnormally expressed in breast cancer, but whether TSP50 is expressed in colorectal carcinoma (CRC) and its clinical significance is unclear. We aimed to detect TSP50 expression in CRC, correlate it with clinicopathological factors, and assess its potential diagnostic and prognostic value. = 0.009).Our data demonstrate that TSP50 is a potential effective indicator of poor survival for CRC patients, especially for those with early-stage tumors