Development of composite amine functionalized polyester microspheres for efficient CO2 capture.

In order to reduce the impact of greenhouse gases on the environment, the development of various new CO2 capture materials has become a hot spot. In this work, a novel composite amine solid adsorbent was prepared by simultaneously using tetraethylenepentamine (TEPA) and 2-[2-(dimethylamino) ethoxy] ethanol (DMAEE) for amine functionalization on the polyester microsphere carrier. The introduction of methyl methacrylate (MMA) with high glass transition temperature into the polyester carrier makes the carrier microspheres have high hardness. At the same time, the carrier also contains active epoxy groups and hydrophobic glycidyl methacrylate (GMA, which can undergo ring-opening reaction with composite amines to achieve high-load and low-energy chemical grafting of amines on the carrier. The composite aminated polyester microspheres were used as an efficient adsorbent for CO2 in simulated flue gas. The results show that the synergistic effect of TEPA-DMAEE composite amine system in the adsorbent is beneficial to the improvement of CO2 capture capacity. When the total amine content in the impregnating solution is 45 wt% and the composite amine ratio is TEPA: DMAEE = 6: 4, the CO2 adsorption capacity can reach the optimal value of 2.45 mmol/ g at 70 °C. In addition, the composite amine microsphere adsorbent has cyclic regeneration performance. Importantly, through kinetic fitting, the Avrami kinetic model fits the CO2 adsorption better than the quasi-first-order and quasi-second-order kinetic models, which proves that physical adsorption and chemical adsorption coexist in the adsorption process. This simple, long-term stable and excellent selective separation performance makes amine-functionalized adsorbents have potential application prospects in CO2 capture.

Tetraethylenepentamine-derived carbon dots and tetraethylenepentamine co-immobilized silica stationary phase for hydrophilic interaction chromatography.

In this work, tetraethylenepentamine (TEPA) was used as precursor and reaction medium to prepare tetraethylenepentamine-functionalized carbon dots (TEPACDs), the resultant mixture was subsequently silanized and then grafted on the surface of bare silica. The obtained tetraethylenepentamine-functionalized carbon dots and tetraethylenepentamine co-modified silica stationary phase (Sil-TEPA/CDs) was characterized by multiple ways, such as Fourier transformed infrared spectroscopy (FTIR), elemental analysis and transmission electron microscope, which revealed the successful preparation of the mixed stationary phase and higher density of functional groups on co-modified stationary phase than precursor single-modified stationary phase. The synergistic effect of TEPACDs and TEPA was proved by comparing the separation performance of Sil-TEPA/CDs and Sil-TEPA toward amino acids, nucleosides, and nucleobases, which distinctly enhanced the selectivity of Sil-TEPA/CDs. Thus, 12 nucleosides and nucleobases and 11 amino acids was nicely separated on Sil-TEPA/CDs. By study the influences of the changes of mobile phase composition, mobile phase buffer concentration and buffer pH on the retention behaviors of Sil-TEPA and Sil-TEPA/CDs, it was found that both hydrophilic partitioning and adsorption of analytes on Sil-TEPA/CDs were enhanced benefit from the co-existence of TEPA and TEPACDs, which provided the analytes better separation performance. By comparing the column quality of Sil-TEPA/CDs with four commercially available columns, Sil-TEPA/CDs exhibited the best peak asymmetry of 0.98, and second best column efficiency of 43895 m-1 using guanosine as analyte. The RSD (n = 9) of the retention times of five selected analytes on Sil-TEPA/CDs were within 0.30-0.61% during 40 h of continuously elution, which implied excellent stability of prepared packing material.

CO2 adsorption and CO2/CH4 separation using fibrous amine-containing adsorbents: isothermal, kinetic, and thermodynamic behaviours.

A series of fibrous aminated adsorbents for CO2 adsorption were prepared by covalent incorporation of poly (glycidyl methacrylate) (PGMA) by graft copolymerization of GMA onto electron beam (EB) irradiated polyethylenepolypropylene (PE/PP) fibrous sheets and subsequent amination with ethylenediamine (EDA), diethylenetriamine (DETA), or tetraethylenepentamine (TEPA). The physico-chemical properties of the adsorbents were evaluated using Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), thermogravimetric (TGA), X-ray diffraction (XRD), and Brunauer-Emmett-Teller (BET) analysis. All the adsorbents displayed typic primary and secondary amine features combined with a decrease in both of crystallinity and surface area of PE/PP, and such a decrease was higher in adsorbents with longer aliphatic chain of the amine. Of all adsorbents, TEPA-containing fibres showed the highest CO2 adsorption capacity and thus was further investigated for CO2 capture from CO2/CH4 mixtures of different gas ratios under various pressures and temperatures. The selectivity of CO2 over CH4 and equilibrium isotherms, kinetics, and thermodynamics of the adsorption on the fibrous aminated adsorbent were all investigated. The Sips model was found to best fit the isotherm of CO2 adsorption suggesting the presence of a combination of monolayer and multilayer adsorptions. The adsorption kinetic data was found to best fit Elovich model reflecting chemisorption. The ΔG°, ΔS°, and ΔH° showed positive values suggesting that the adsorption of CO2 on the present fibrous adsorbent was non-spontaneous with an increase in randomness implying that the process was endothermic. Overall, it can be suggested that PE/PP-g-PGMA/TEPA adsorbent has a strong potential for separation of CO2 from NG.

Design of reversibly charge-changeable rhodamine B modified magnetic nanoparticles to enrich phosphopeptides.

In the present study, by employing ethylenediaminetetraacetic acid (EDTA), tetraethylene pentaamine (TEPA), and rhodamine B (Rb), we designed and synthesized a magnetic adsorbent (Fe3O4@EDTA@TEPA@Rb) on the basis of reversible charge change of Rb and applied to capture phosphopeptides. Rb existing in open planarized zwitterion form when stimulated by acidic loading buffer adsorbs negative phosphopeptides via electrostatic interaction. Under the stimulation of alkalic eluent, ring-closed structure of Rb is formed to elute the enriched phosphopeptides. TEPA containing rich amino groups is used as a crosslinking agent, which is also protonated in acidic loading buffer to bond phosphopeptides. Then phosphopeptides are eluted when TEPA deprotonates in alkalic eluent. Coupled with matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) detection, phosphopeptide signals originated from 0.4 fmol/µL ß-casein digests were successfully detected. In addition, Fe3O4@EDTA@TEPA@Rb can also efficiently enrich phosphopeptides from skimmed milk, human serum and saliva samples (26, 4, 39 phosphopeptides, respectively), opening a new gallery for phosphopeptides-related analysis. In general, the developed adsorbent has the great potential for further application in the near future.

Therapeutic education and physical activity are feasible and safe in hematologic cancer patients referred to chemotherapy: results of a randomized controlled trial.

PURPOSE: Although over 60% of patients with hematologic cancer report distressing fatigue, they often do not receive recommendations on fatigue management strategies. The aim of this pilot study was to estimate the feasibility of therapeutic education and physical activity (TEPA) by measuring the patients' adherence to this multidimensional intervention. The secondary aim was to estimate the impact of TEPA on clinical outcomes. METHODS: Patients with hematologic cancer participated in this single-center, open-label, randomized controlled trial. The control group (CG) received two educational group sessions on fatigue and physical activity. The experimental group (EG) received the two educational sessions plus six weekly individual sessions aimed at implementing a personalized physical exercise program. Follow-ups were at 1, 3, and 7 months. RESULTS: Forty-six patients referred to chemotherapy were included, corresponding to 54% of recruitment rate. Adherence reached 90% in the EG and 68% in the CG. Most patients (65% in EG and 64% in CG) attended a minimum of 80% of the planned sessions. Overall retention rate was 87% (85% in EG and 91% in CG). No adverse events were registered. No between-group differences were detected in fatigue (FACIT-F), psychological distress (NCCN Distress Thermometer), QoL (EORTC QLQ-C30), or functional exercise capacity (TUG test and 6MWT). Adherence to an active lifestyle, measured by a semi-structured interview, increased from 56.5 to 84% in the EG at 7 months (p = 0.02), whereas it decreased slightly in the CG (from 47.8 to 42.9%). CONCLUSION: Multidimensional rehabilitation interventions are feasible and safe in this population, and larger trials should focus on the efficacy of such approaches on clinically relevant outcomes. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT03403075.

Removal of Cr(VI) and Ag(I) by grafted magnetic zeolite/chitosan for water purification: Synthesis and adsorption mechanism.

In this paper, a novel grafted zeolite/Fe3O4/chitosan (ZMC-MAH-TEPA) adsorbent was greenly synthesized and evaluated for the removal of Cr(VI) and Ag(I) in single and bi-component solutions. The characterization data of XRF, XRD, FT-IR, VSM, TGA, BET and SEM showed the successful fabrication of the adsorbent with abundant -NH2 and -NH- as well as great recovery properties (11.70 emu/g). The effects of experimental parameters, including pH value, initial concentration, temperature, time, and coexisting ions on single and bi-component adsorption, were investigated. The results demonstrated that the adsorption capacities increased with the enhanced shaking time and concentration until equilibrium was reached. The optimum pH value was 3 for Cr(VI) adsorption and 5 for Ag(I) adsorption. The maximum adsorption capacities for Cr(VI) and Ag(I) ions, obtained by the Langmuir model, were 50.75 and 70.12 mg·g-1 in single metal solutions and 45.45 and 58.94 mg·g-1 in bi-component metal solutions (Cr(VI)/Ag(I) = 1:1), respectively. Additionally, the adsorption mechanism of Cr(VI) and Ag(I) was explained in terms of electrostatic interaction and hydrogen bonding between metal ions and -NH2 and -NH- in ZMC-MAH-TEPA. Taken together, this study provides a clean approach to synthesizing chitosan-based adsorbents for the efficient removal of Cr(VI) and Ag(I) ions.

Cytotoxic Effects Caused by Functionalized Carbon Nanotube in Murine Macrophages.

BACKGROUND/AIMS: The development of new nanomaterials has been growing in recent decades to bring benefits in several areas, especially carbon-based nanoparticles, which have unique physical-chemical properties and allow to take on several applications. Consequently, the use of new nanomaterials without previous toxicological studies raises concern about possible harmful health effects. The aim of this study was to investigate the cytotoxic profile of a new multi-walled carbon nanotube (MWCNT) functionalized with tetraethylenepentamine called OCNT-TEPA using in vitro assays in murine macrophage cells linage J774 A.1. METHODS: OCNT-TEPA was characterized by transmission electron microscopy (TEM) and high resolution TEM (HR-TEM), scanning electron microscopy (SEM), zeta potential and dynamic light scattering (DLS), and its cytotoxic effects were evaluated at 24 and 48 hours by cell viability assays (MTT and NR), morphology and cell recovery (optic microscopy and clonogenic assay), formation of reactive oxygen (ROS) and nitric oxide (NO) species, inflammatory profile (IL-6 and TNF cytokines), mitochondrial membrane potential analysis (MMP), activation of the caspase 3 pathway and cell death (flow cytometry). RESULTS: The data showed a significant decrease in cell viability, increased production of ROS and NO, alteration of mitochondrial membrane potential, increased levels of inflammatory cytokines, alteration of cell morphology, activation of the Caspase 3 pathway and consequently cell death, in the highest concentrations of OCNT-TEPA tested in the periods of 24 and 48 hours. CONCLUSION: The analyses showed that OCNT-TEPA has a dose-dependent cytotoxic profile, which may be harmful to murine macrophages (J774 A.1) and may represent a health risk.

Directional spatial processes override non-directional ones in structuring communities of lotic macroinvertebrates differing in dispersal ability.

Dispersal is an essential natural process that influences community assembly, yet directional dispersal through wind and water may have distinctive effects. Environmental and spatial factors jointly influence community structure, but their relative importance is anticipated to vary with spatial distance, dispersal mode, and season. Accordingly, a systemic survey was conducted in subtropical Chinese mountain lotic systems to distinguish the relative contributions of environmental control and spatial structuring upon communities of macroinvertebrates with different dispersal ability. Macroinvertebrate samples were collected from the upper reaches and five tributaries of the Hanjiang River in October 2017 (autumn) and April 2018 (spring). These macroinvertebrates were identified and classified into three dispersal groups: aquatic passive (AqPa), terrestrial passive (TePa), and terrestrial active (TeAc). Variation partitioning analyses were performed on environmental factors and different sets of spatial factors (overland dispersal: Overland, directional downwind dispersal: AEM_Wind, along watercourse dispersal: Watercourse, and directional downstream dispersal: AEM_Water). Findings showed that both environmental filtering and spatial structuring influenced the structure of macroinvertebrate metacommunities. For AqPa and TePa groups, pure environmental effects were stronger than pure spatial effects based on most distance matrices; however, in AEM_Water, the effects of spatial processes surpassed those of environmental filtering. For TeAc group, the role of environmental control and spatial structuring varied depending on different spatial models. The results also highlighted seasonal shifts in metacommunity structuring processes. Spatial structures featuring direction, especially AEM_Water, were predominant in explaining the construction of macroinvertebrate communities. This work suggests that directional dispersal should be explicitly considered when examining the structure of ecological communities.

Synthesis and application of 5 µm monodisperse porous silica microspheres with controllable pore size using polymeric microspheres as templates for the separation of small solutes and proteins by high-performance liquid chromatography.

High-performance liquid chromatography (HPLC) is a powerful tool to separate and analyze complex samples. Monodiseperse porous silica microspheres (MPSMs) have been widely used as column packings in HPLC. However, synthesis of MPSMs with controllable sizes of both particles and pores for the separation of small solutes and proteins in HPLC still remains a challenge. In this paper, an effective and facile approach to prepare MPSMs with controllable particle size and pore size by using porous polymer microspheres as templates is presented. By employing porous PGMA/EDMA microspheres as templates and tetraethyl orthosilicate (TEOS) as the silica source, 5 µm MPSMs with tunable pore sizes were synthesized successfully. The as-prepared MPSMs were characterized by scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy, thermogravimetric analysis (TGA), dynamic laser scattering, and mercury intrusion porosimetry. The results indicated that the MPSMs obtained retained the original size of the polymer templating particles and calcination caused almost no shrinkage. Furthermore, the effects of the pore size of polymer template microspheres, different amino-functionalizations of PGMA/EDMA microspheres and the mass ratio of PGMA/EDMA microspheres/TEOS on the pore size of MPSMs were carefully studied. The results indicated that the pore size of MPSMs was adjusted from 20 to 69 nm by controlling the pore size of the polymer microspheres and the mass ratio of PGMA/EDMA microspheres/TEOS in the sol-gel process. In addition, the amino-functionalization of PGMA/EDMA microspheres with different structure-directing agents, such as (3-aminopropyl)triethoxysilane (APTES), trimethylamine hydrochloride (TMA) and tetraethylenepentamine (TEPA), also resulted in MPSMs with the different pore sizes. MPSMs with large pore sizes of more than 30 nm were fabricated by using TEPA-functionalized PGMA/EDMA microspheres as templates, while with TMA-functionalized PGMA/EDMA microspheres as templates, MPSMs with pore sizes of approximately 10 nm were obtained. The as-prepared MPSMs were further modified with different silanes, such as C4, C8 and C18, to explore as stationary phases for the separation of proteins and small solutes in reversed phase liquidi chromatography (RPLC). The results illustrated that the baseline separation of 7 kinds of proteins could be achieved based on MPSMs with pore sizes of 30 nm, and 6 kinds of alkyl benzenes and 5 kinds of aromatic alcohol homologs could be separated completely based on MPSMs with pore sizes of 11 nm. This work demonstrated that MPSMs prepared by applying the polymer templating method showed a promising potential applicability in HPLC.

To Transect or Not Transect: Results from the Scandinavian Urethroplasty Study, A Multicentre Randomised Study of Bulbar Urethroplasty Comparing Excision and Primary Anastomosis Versus Buccal Mucosal Grafting.

BACKGROUND: Open surgical treatment of short bulbar urethral strictures (urethroplasty) is commonly performed as transecting excision and primary anastomosis (tEPA) or buccal mucosa grafting (BMG). Erectile dysfunction and penile complications have been reported, but there is an absence of randomised trials. OBJECTIVE: To evaluate sexual dysfunction and penile complications after urethroplasty with tEPA versus BMG. DESIGN, SETTING, AND PARTICIPANTS: Centres in Finland, Sweden and Norway participated. Patients with a bulbar urethral stricture of ≤2 cm without previous urethroplasty were randomised. The primary endpoints were the degree of erectile dysfunction and penile complications. Follow-up was 12 mo. INTERVENTION: Patients were randomised to either tEPA or BMG urethroplasty. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Sexual dysfunction was measured using the International Index of Erectile Function, 5-item version (IIEF-5) and a penile complications questionnaire (PCQ) designed for this study. Continuous data were analysed using analysis of covariance and categorical data were compared using a χ2 test. RESULTS AND LIMITATIONS: A total of 151 patients were randomised to either tEPA (n = 75) or BMG (n = 76). The tEPA group reported more penile complications (p = 0.02), especially reduced glans filling (p = 0.03) and a shortened penis (p = 0.001). There were no differences in postoperative IIEF-5 total scores. Recurrence rates were similar in both groups (12.9%) but the study was not designed to detect differences in recurrence rates. The PCQ is not validated, which is a limitation. CONCLUSIONS: More patients reported penile complications after urethroplasty with tEPA than with BMG. This should be considered when choosing the operative method, and patients should be informed accordingly. PATIENT SUMMARY: This study compared two common operations for repair of narrowing of the male urethra. Neither of the two methods seems to cause worsened erections. However, penile problems are more common after the transection technique than after the grafting technique.

First-principles studies on two-dimensional B3O3 adsorbent as a potential drug delivery platform for TEPA anticancer drug.

The remarkable properties of pristine B3O3 nanosheet as a nanocarrier for adsorption and desorption of TEPA anticancer drug for designing potential drug delivery platform were investigated using periodic DFT calculations. We studied the adsorption energy of all stable complexes formed between the drug molecule and B3O3 in gas and aqueous phases along with electronic structure analysis of complexes. Different adsorption configurations were studied for drug/B3O3 complexes, including the interaction of the C atom of the triangular ring, O atom in the TEPA drug with the B atom in B3O3, and indirect drug interaction the middle of the R1 ring cavity of the B3O3 nanosheet. The take-up of TEPA prompts a substantial change of 68.13% in the band gap (Eg) of the B3O3 nanosheet in the most stable complex. The present study results affirmed the application of B3O3 nanosheet as a potential vehicle for TEPA drugs in the treatment of cancerous tissues.

Intratumoral Copper Modulates PD-L1 Expression and Influences Tumor Immune Evasion.

Therapeutic checkpoint antibodies blocking programmed death receptor 1/programmed death ligand 1 (PD-L1) signaling have radically improved clinical outcomes in cancer. However, the regulation of PD-L1 expression on tumor cells is still poorly understood. Here we show that intratumoral copper levels influence PD-L1 expression in cancer cells. Deep analysis of the The Cancer Genome Atlas database and tissue microarrays showed strong correlation between the major copper influx transporter copper transporter 1 (CTR-1) and PD-L1 expression across many cancers but not in corresponding normal tissues. Copper supplementation enhanced PD-L1 expression at mRNA and protein levels in cancer cells and RNA sequencing revealed that copper regulates key signaling pathways mediating PD-L1-driven cancer immune evasion. Conversely, copper chelators inhibited phosphorylation of STAT3 and EGFR and promoted ubiquitin-mediated degradation of PD-L1. Copper-chelating drugs also significantly increased the number of tumor-infiltrating CD8+ T and natural killer cells, slowed tumor growth, and improved mouse survival. Overall, this study reveals an important role for copper in regulating PD-L1 and suggests that anticancer immunotherapy might be enhanced by pharmacologically reducing intratumor copper levels. SIGNIFICANCE: These findings characterize the role of copper in modulating PD-L1 expression and contributing to cancer immune evasion, highlighting the potential for repurposing copper chelators as enhancers of antitumor immunity. GRAPHICAL ABSTRACT: http://cancerres.aacrjournals.org/content/canres/80/19/4129/F1.large.jpg.

A novel sandwich aptasensor for detecting T-2 toxin based on rGO-TEPA-Au@Pt nanorods with a dual signal amplification strategy.

T-2 toxin is a mycotoxin that can cause chronic illnesses, and the detection of T-2 toxin in food is critical for human health. Herein, a novel sandwich aptasensor with a dual signal amplification strategy was developed for the detection of T-2 toxin. Molybdenum disulfide-polyaniline-chitosan-gold nanoparticles (MoS2-PANI-Chi-Au) were processed to the modified glassy carbon electrode (GCE) and used as the aptasensor platform to expedite the electronics transport and immobilize the amino-terminated capture DNA probe by Au-N bonds. The reduced graphene oxide-tetraethylene pentamine-gold@platinum nanorods (rGO-TEPA-Au@Pt NRs) were first synthesized and immobilized with a signal DNA probe. Once T-2 toxin was added into the biosensing system, the aptamer would trap T-2 toxin to turn the signal off. Next, dissociative aptamer hybridized with the capture DNA probe in GCE and linked simultaneously to the signal DNA probe on rGO-TEPA-Au@Pt NRs with another end sequence of aptamer to turn the signal on. Owing to the efficient catalytic ability of bimetallic Au@Pt nanorods, the signal was perfectly amplified through the catalysis of hydrogen peroxide (H2O2) and recorded by chronoamperometry. With the outstanding augment response, the limit of detection reached 1.79 fg mL-1 (3SB/m) and a wide linear range from 10 fg mL-1 to 100 ng mL-1 was presented. The sensitivity of the aptasensor was 19.88 µA⋅µM-1⋅cm-2. Meanwhile, the DNA aptamer-bimetallic nanorod based sensing system presented excellent specificity. The developed aptasensor provides a new platform for T-2 toxin detection with low cost for real sample assays.

Transformation of alkylating regimen of thiotepa into tepa determines the disease progression through GSTP1 gene polymorphism for metastatic breast cancer patients receiving thiotepa containing salvage chemotherapy.

BACKGROUND: The shifts to second-line chemotherapy for metastatic breast cancer (MBC) were widely required based on pharmaceutical molecular profiles to reach out precision medicine. The emerging precise treatment of cancer requires the implementation of clarified pharmacogenetic profiles which are capable of elucidating the predictive responses to cancer chemotherapy. Therefore we were interested in the analysis of the roles of single nucleotide polymorphism (SNP) of GSTP1 (glutathione S-transferase pi 1 gene) alleles to identify pharmacological links with predictors of clinical responses and toxicities. METHODS: 93 MBC patients receiving thiotepa plus docetaxel chemotherapy were enrolled in this study. Optimized CYP3A5, CYP2B6, and GSTP1 were predominantly selected as candidate genes and their three SNPs (CYP2B6 G516T, CYP3A5 A6986G, and GSTP1 A313G) were genotyped by matrix-assisted laser desorption ionization/time of flight (MALDI-TOF) mass spectrometry. Progression-free survival (PFS), disease control rate, and chemo-related toxicities were recorded. RESULTS: GSTP1 A313G (rs1695) was identified to be related with disease progression. In particular, patients harboring AG/GG genotype demonstrated a statistically longer PFS than those with AA. Multivariate analysis confirmed that AG/GG genotype was associated with both clinical responses and liver-localized metastatic lesions. No correlation was found between these three SNPs and chemotherapy-induced toxicity. CONCLUSIONS: These results suggest that the GSTP1 polymorphism is a novel prognostic marker for clinical response to thiotepa-containing chemotherapy regimens. Such evidence could provide insight into the role of pharmacogenetics to deprive of biases in shifting regimens solely by empirical choices.

Copper depletion inhibits CoCl2-induced aggressive phenotype of MCF-7 cells via downregulation of HIF-1 and inhibition of Snail/Twist-mediated epithelial-mesenchymal transition.

Copper, a strictly regulated trace element, is essential for many physiological processes including angiogenesis. Dysregulated angiogenesis has been associated with increased copper in tumors, and thus copper chelators have been used to inhibit tumor angiogenesis. However, it remains unclear whether copper has any effect on epithelial-mesenchymal transition (EMT). Using CoCl2-induced EMT of human breast carcinoma MCF-7 cells, we found that TEPA, a copper chelator, inhibited EMT-like cell morphology and cytoskeleton arrangement triggered by CoCl2; decreased the expression of vimentin and fibronectin, markers typical of EMT; inhibited HIF-1 activation and HIF1-α accumulation in nuclear; and down-regulated the expression of hypoxia-associated transcription factors, Snail and Twist1. Moreover, knockdown copper transport protein, Ctr1, also inhibited CoCl2-induced EMT and reversed the mesenchymal phenotype. In EMT6 xenograft mouse models, TEPA administration inhibited the tumor growth and increased mice survival. Immunohistochemical analysis of the xenograft further demonstrated that TEPA administration significantly inhibited tumor angiogenesis, down-regulated hypoxia-induced transcription factors, Snail and Twist1, leading to decreased transactivation of EMT-associated marker genes, vimentin and fibronectin. These results indicate that TEPA inhibits CoCl2-induced EMT most likely via HIF1-α-Snail/Twist signaling pathway, and copper depletion may be exploited as a therapeutic for breast cancer.

PET Imaging of Macrophage Mannose Receptor-Expressing Macrophages in Tumor Stroma Using 18F-Radiolabeled Camelid Single-Domain Antibody Fragments.

UNLABELLED: Tumor-associated macrophages constitute a major component of the stroma of solid tumors, encompassing distinct subpopulations with different characteristics and functions. We aimed to identify M2-oriented tumor-supporting macrophages within the tumor microenvironment as indicators of cancer progression and prognosis, using PET imaging. This can be realized by designing (18)F-labeled camelid single-domain antibody fragments (sdAbs) specifically targeting the macrophage mannose receptor (MMR), which has been identified as an important biomarker on this cell population. METHODS: Cross-reactive anti-MMR sdAbs were generated after immunization of an alpaca with the extracellular domains of both human and mouse MMR. The lead binder was chosen on the basis of comparisons of binding affinity and in vivo pharmacokinetics. The PET tracer (18)F-fluorobenzoate (FB)-anti-MMR sdAb was developed using the prosthetic group N-succinimidyl-4-(18)F-fluorobenzoate ((18)F-SFB), and its biodistribution, tumor-targeting potential, and specificity in terms of macrophage and MMR targeting were evaluated in mouse tumor models. RESULTS: Four sdAbs were selected after affinity screening, but only 2 were found to be cross-reactive for human and mouse MMR. The lead anti-MMR 3.49 sdAb, bearing an affinity of 12 and 1.8 nM for mouse and human MMR, respectively, was chosen for its favorable in vivo biodistribution profile and tumor-targeting capacity. (18)F-FB-anti-MMR 3.49 sdAb was synthesized with a 5%-10% radiochemical yield using an automated and optimized protocol. In vivo biodistribution analyses showed fast clearance via the kidneys and retention in MMR-expressing organs and tumor. The kidney retention of the fluorinated sdAb was 20-fold lower than a (99m)Tc-labeled counterpart. Compared with MMR- and C-C chemokine receptor 2-deficient mice, significantly higher uptake was observed in tumors grown in wild-type mice, demonstrating the specificity of the (18)F tracer for MMR and macrophages, respectively. CONCLUSION: Anti-MMR 3.49 was denoted as the lead cross-reactive MMR-targeting sdAb. (18)F radiosynthesis was optimized, providing an optimal probe for PET imaging of the tumor-promoting macrophage subpopulation in the tumor stroma.

Evaluation of umbilical cord blood CD34 (+) hematopoietic stem cell expansion in co-culture with bone marrow mesenchymal stem cells in the presence of TEPA.

BACKGROUND: During the last three decades hematopoietic stem cells (HSC) have become a standard protocol for the treatment of many hematologic malignancies and non-malignant disorders. Umbilical cord blood (UCB), as a source of HSCs, has many advantages compared with other sources. One major drawback in using this source in treatment of adult patients is the low HSC dose available. Ex vivo expansion of HSCs is a solution to overcome this limitation. In this study we used TEPA, as a Cu chelator, and human bone marrow (BM) mesenchymal stem cells (MSCs) to investigate expansion rate of UCB-HSCs. MATERIALS AND METHODS: CB-HSCs were isolated using miniMACS magnetic separation system. We cultured the enriched CD34(+)cells in various conditions: culture condition A, supplemented only with recombinant cytokines; culture condition B, supplemented with BM-MSCs as a cell feeder layer and recombinant cytokines; culture condition C, supplemented with recombinant cytokines and TEPA; culture condition D, supplemented with recombinant cytokines, BM-MSCs as a cell feeder layer and TEPA. In order to evaluate the HSC expansion, we performed cell count, analysis of CD34(+) expression by flow cytometry, and colony-forming cell assay on Day 10 after culture. RESULTS: The most fold increase in CD34(+) cell, total cell, and total colony numbers was observed in culture condition D (110.11 ± 15.3, 118.5 ± 21, and 172.9 ± 44.7, respectively) compared to other conditions. CONCLUSION: The results showed that co-culture of HSCs with BM-MSCs in the presence of copper chelating agent (TEPA) could dramatically increase expansion rate of UCB-HSCs. Therefore, this strategy could be useful for HSC expansion.

Mechanisms and kinetics of thiotepa and tepa hydrolysis: DFT study.

N,N',N″-triethylenethiophosphoramide (Thiotepa) and its oxo analogue (Tepa) as the major metabolite are trifunctional alkylating agents with a broad spectrum of antitumor activity. In vivo and vitro studies show alkylation of DNA by Thiotepa and Tepa can follow two pathways, but it remains unclear which pathway represents the precise mechanism of action. In pathway 1, these agents are capable of forming cross-links with DNA molecules via two different mechanisms. In the first mechanism, the ring opening reaction is initiated by protonating the aziridine, which then becomes the primary target of nucleophilic attack by the N7-Guanine. The second one is a direct nucleophilic ring opening of aziridyl group. Thiotepa and Tepa in pathway 2, act as a cell penetrating carrier for aziridine, which is released via hydrolysis. The released aziridine can form a cross-link with N7-Guanine. In this study, we calculated the activation free energy and kinetic rate constant for hydrolysis of these agents and explored interaction of aziridine with Guanine to predict the most probable mechanism by applying density functional theory (DFT) using B3LYP method. In addition, solvent effect was introduced using the conductor-like polarizable continuum model (CPCM) in water, THF and diethylether. Hyperconjugation stabilization factors that have an effect on stability of generated transition state were investigated by natural bond order (NBO) analysis. Furthermore, quantum theory of atoms in molecules (QTAIM) analysis was performed to extract the bond critical points (BCP) properties, because the electron densities can be considered as a good description of the strength of different types of interactions.

Polymorphisms of drug-metabolizing enzymes (GST, CYP2B6 and CYP3A) affect the pharmacokinetics of thiotepa and tepa.

AIMS: Thiotepa is widely used in high-dose chemotherapy. Previous studies have shown relations between exposure and severe organ toxicity. Thiotepa is metabolized by cytochrome P450 and glutathione S-transferase enzymes. Polymorphisms of these enzymes may affect elimination of thiotepa and tepa, its main metabolite. The purpose of this study was to evaluate effects of known allelic variants in CYP2B6, CYP3A4, CYP3A5, GSTA1 and GSTP1 genes on pharmacokinetics of thiotepa and tepa. METHODS: White patients (n = 124) received a high-dose regimen consisting of cyclophosphamide, thiotepa and carboplatin as intravenous infusions. Genomic DNA was analysed using polymerase chain reaction and sequencing. Plasma concentrations of thiotepa and tepa were determined using validated GC and LC-MS/MS methods. Relations between allelic variants and elimination pharmacokinetic parameters were evaluated using nonlinear mixed effects modelling (nonmem). RESULTS: The polymorphisms CYP2B6 C1459T, CYP3A4*1B, CYP3A5*3, GSTA1 (C-69T, G-52A) and GSTP1 C341T had a significant effect on clearance of thiotepa or tepa. Although significant, most effects were generally not large. Clearance of thiotepa and tepa was predominantly affected by GSTP1 C341T polymorphism, which had a frequency of 9.3%. This polymorphism increased non-inducible thiotepa clearance by 52% [95% confidence interval (CI) 41, 64, P < 0.001] and decreased tepa clearance by 32% (95% CI 29, 35, P < 0.001) in heterozygous patients, which resulted in an increase in combined exposure to thiotepa and tepa of 45% in homozygous patients. CONCLUSIONS: This study indicates that the presently evaluated variant alleles explain only a small part of the substantial interindividual variability in thiotepa and tepa pharmacokinetics. Patients homozygous for the GSTP1 C341T allele may have enhanced exposure to thiotepa and tepa.

Population pharmacokinetics of cyclophosphamide and its metabolites 4-hydroxycyclophosphamide, 2-dechloroethylcyclophosphamide, and phosphoramide mustard in a high-dose combination with Thiotepa and Carboplatin.

The anticancer prodrug cyclophosphamide (CP) is activated by the formation of 4-hydroxycyclophosphamide (4OHCP), which decomposes into phosphoramide mustard (PM). This activation pathway is inhibited by thiotepa. CP is inactivated by formation of 2-dechloroethylcyclophosphamide (2DCECP). The aim of this study was to develop a population pharmacokinetic model describing the complex pharmacokinetics of CP, 4OHCP, 2DCECP, and PM when CP is administered in a high-dose combination with thiotepa and carboplatin. Patients received a combination of CP (1000-1500 mg/m/d), carboplatin (265-400 mg/m/d), and thiotepa (80-120 mg/m/d) administered in short infusions over 4 days. Twenty blood samples were collected per patient per course. Concentrations of CP, 4OHCP, 2DCECP, PM, thiotepa, and tepa were determined in plasma. Using NONMEM, an integrated population pharmacokinetic model was used to describe the pharmacokinetics of CP, 4OHCP, 2DCECP, and PM, including the already described processes of autoinduction of CP and the interaction with thiotepa. Data were available on 35 patients (70 courses). The pharmacokinetics of CP were described with a 2-compartment model, and those of 4OHCP, 2DCECP, and PM with 1-compartment models. Before onset of autoinduction, it was assumed that CP is eliminated through a noninducible pathway accounting for 20% of total CP clearance, whereas 2 inducible pathways resulted in formation of 4OHCP (75%) and 2DCECP (5%). It was assumed that 4OHCP was fully converted to PM. Induction of CP metabolism was mediated by 2 hypothetical amounts of enzyme whose quantities increased in time in the presence of CP (kenz=0.0223 and 0.0198 hours). Induction resulted in an increased formation of 4OHCP (approximately 50%), PM (approximately 50%), and 2DCECP (approximately 35%) during the 4-day course, and concomitant decreased exposure to CP (approximately 50%). The formation of 2DCECP was not inhibited by thiotepa. Apparent volumes of distribution of CP, PM, and 2DCECP could be estimated being 43.7, 55.5, and 18.5 L, respectively. Exposure to metabolites varied up to 9-fold. The complex population pharmacokinetics of CP, 4OHCP, 2DCECP, and PM in combination with thiotepa and carboplatin has been established and may form the basis for further treatment optimization with this combination.