Gelatin/PLLA sponge-like scaffolds allow proliferation and osteogenic differentiation of human mesenchymal stromal cells

Tissue engineering has the potential to supply constructs capable of restoring the normal function of native tissue following injury. Poly(L-lactic acid) (PLLA) scaffolds are amongst the most commonly used biodegradable polymers in tissue engineering and previous studies performed on ovine fibroblasts have showed that addition of gelatin creates a favorable hydrophilic microenvironment for the growth of these cells. The attractiveness of using mesenchymal stromal cells (MSCs) in tissue regeneration is that they are able to differentiate into several lines including osteoblasts. In this study, we investigated the ability of gelatin/PLLA sponges to support the adhesion, proliferation, and osteogenic differentiation of human MSCs isolated from the bone marrow of four donors. [Figure: see text].     

[161Tb]Tb-Thz-Phe-D-Trp-Lys-Thr-DOTA: A potential radiopharmaceutical for the treatment of neuroendocrine tumors

Thz-Phe-D-Trp-Lys-Thr-DOTA, a conjugate of the DOTA chelator and the Thz-Phe-D-Trp-Lys-Thr pentapeptide, was labeled with ¹⁵²Eu and ¹⁶¹Tb radionuclides, where ¹⁶¹Tb has decay characteristics suitable for its use in cancer therapy. For the [¹⁵²Eu]Eu-Thz-Phe-D-Trp-Lys-Thr-DOTA complex, the biodistribution in nude mice bearing IMR-32 tumors was evaluated for the first time. It was shown that the complexes of the conjugate demonstrate accumulation in the tumor at the level of DOTA-TATE, another peptide conjugate widely used in nuclear medicine for the diagnosis and therapy of neuroendocrine tumors, which allows Thz-Phe-D-Trp-Lys-Thr-DOTA to be considered as a potential biological vector for radiopharmaceuticals.     

New Oxaliplatin-Pyrophosphato Analogs with Improved In Vitro Cytotoxicity

Two new Pt(II)-pyrophosphato complexes containing the carrier ligands cis-1,3-diaminocyclohexane (cis-1,3-DACH) and trans-1,2-diamine-4-cyclohexene (1,2-DACHEX), variants of the 1R,2R-diaminocyclohexane ligand present in the clinically used Pt-drug oxaliplatin, have been synthesized with the aim of developing new potential antitumor drugs with high bone tropism. The complexes are more stable at physiological pH than in acid conditions, with Na₂[Pt(pyrophosphato)(cis-1,3-DACH)] (1) slightly more stable than [Pt(dihydrogenpyrophosphato)(1,2-DACHEX)] (2). The greater reactivity at acidic pH ensures a greater efficacy at the tumor site. Preliminary NMR studies indicate that 1 and 2 react slowly with 5’-GMP (used as a model of nucleic acids), releasing the pyrophosphate ligand and affording the bis 5’-GMP adduct. In vitro cytotoxicity assays performed against a panel of four human cancer cell lines have shown that both compounds are more active than oxaliplatin. Flow cytometry studies on HCT116 cells showed that the pyrophosphato compounds with the non-classical 1,3- and 1,4-diaminocyclohexane ligands (1 and 4) are the most capable to induce cells’ death by apoptosis and necrosis.     

Magnetic resonance imaging measurements of vascular permeability and extracellular volume fraction of breast tumors by dynamic Gd-DTPA-enhanced relaxometry

Vascular permeability (k(ep), min(-1)) and extracellular volume fraction (v(e)) are tissue parameters of great interest to characterize malignant tumor lesions. Indeed, it is well known that tumors with high blood supply better respond to therapy than poorly vascularized tumors, and tumors with large extracellular volume tend to be more malignant than tumors showing lower extracellular volume. Furthermore, the transport of therapeutic agents depends on both extracellular volume fraction and vessel permeability. Thus, before treatment, these tissue parameters may prove useful to evaluate tumor aggressiveness and to predict responsiveness to therapy and variations during cytotoxic therapies could allow to assess treatment efficacy and early modified therapy schedules in case of poor responsiveness. As a consequence, there is a need to develop methods that could be routinely used to determine these tissue parameters. In this work, blood-tissue permeability and extracellular volume fraction information were derived from magnetic resonance imaging dynamic longitudinal relaxation rate (R(1)) mapping obtained after an intravenous bolus injection of Gd-DTPA in a group of 92 female patients with breast lesions, 68 of these being histologically proven to be with carcinoma. For the sake of comparison, 24 benign lesions were studied. The measurement protocol based on two-dimensional gradient echo sequences and a monoexponential plasma kinetic model was that validated in the occasion of previous animal experiments. As a consequence of neoangiogenesis, results showed a higher permeability in malignant than in benign lesions, whereas the extracellular volume fraction value did not allow any discrimination between benign and malignant lesions. The method, which can be easily implemented whatever the imaging system used, could advantageously be used to quantify lesion parameters (k(ep) and v(e)) in routine clinical imaging. Because of its large reproducibility, the method could be useful for intersite comparisons and follow-up studies.     

Solid-phase extraction and ultra high-performance liquid chromatography tandem mass spectrometry analysis of the gastrointestinal absorption of emodin in different digestive segments of rats

A rapid, simple and sensitive ultra high‐performance liquid chromatography (UHPLC‐MS/MS) method was established for determining the absorption amount of emodin in the five digestive segments of rat. Plasma samples were pre‐purified by solid‐phase extraction (SPE) with Oasis MAX cartridge. Separation of emodin and 1,8‐dihydroxyanthraquinone (internal standard) was performed on an Acquity BEH UHPLC C₁₈ column (100 mm×2.1 mm, 1.7 μm) with gradient elution of methanol and 0.1% formic acid aqueous solution. The LC/MS system was operated under multiple reaction monitoring mode using electrospray ionization (ESI) in negative ion mode. The results showed that this established method was valid and sensitive for emodin within 0.04–16.4 μg/mL, with low limits of detection and quantification of 0.6 ng/mL and 2.4 ng/mL, respectively and upper limit of quantification of 220.0 ng/mL. The intra‐ and interday variations were below 4.9% of RSD. The extraction recoveries were 98.9–106.1% with RSD of 1.9–3.2%. The plasma concentration‐time relationship showed that the absorption of emodin in stomach was faster than in intestine segments. The sequence of absorption amount was: ileum>jejunum>colon≈duodenum>stomach. Most of emodin was absorbed in ileum, and the absorption amount was increased with prolonged retention of drug form in intestine, especially in ileum and jejunum. The developed UHPLC‐ESI‐MS/MS method was appropriate for determining the in vivo absorption of emodin in other herbal medicines or preparations containing this compound.     

傅里叶变换红外光谱技术诊断恶性肿瘤的研究进展

恶性肿瘤严重威胁人类的健康和生命,因此其早期诊断显得尤为重要。近年来,傅里叶变换红外光谱技术在诊断恶性肿瘤方面显示出了巨大的潜力。与传统方法相比,该技术在诊断肿瘤方面具有准确、快速、无创、原位、廉价、自动化、可重复、无需预处理、能够在分子水平上早期诊断等显著优势。本文综述了FTIR技术应用于诊断呼吸、消化、泌尿、生殖、神经、皮肤、血液等各系统恶性肿瘤方面的研究进展,并结合国际上临床医学、光谱学和化学计量学的学科发展现状提出了五点展望：扩大样本量,进行多中心研究;与内窥镜、穿刺活检结合,实现术中实时原位诊断并指导活检;进一步实现自动化;找寻更为高效的化学计量学方法;个别指认比较困难的光谱参数的识别尚有待进一步研究证实。随着FTIR技术的进一步发展和完善,它必将成为一种辅助诊断恶性肿瘤的重要方法,甚至可能作为肿瘤的常规筛查手段并应用于其分期分级中。     

Effective particle methods for Fisher–Kolmogorov equations: Theory and applications to brain tumor dynamics

Extended systems governed by partial differential equations can, under suitable conditions, be approximated by means of sets of ordinary differential equations for global quantities capturing the essential features of the systems dynamics. Here we obtain a small number of effective equations describing the dynamics of single-front and localized solutions of Fisher–Kolmogorov type equations. These solutions are parametrized by means of a minimal set of time-dependent quantities for which ordinary differential equations ruling their dynamics are found. A comparison of the finite dimensional equations and the dynamics of the full partial differential equation is made showing a very good quantitative agreement with the dynamics of the partial differential equation. We also discuss some implications of our findings for the understanding of the growth progression of certain types of primary brain tumors and discuss possible extensions of our results to related equations arising in different modeling scenarios.     

Microcalorimetry for assessment of energy turnover of human gastrointestinal mucosa in vivo

The objective of the present microcalorimetric investigation on mucosal biopsies from human gastric body (B) and descending duodenum (D) was to test whether a steady state of metabolism does persist and, if not, to find out how to assess the preexisting in vivo conditions. The measurements were performed at various incubation temperatures and partial pressures of oxygen. The measured curves fell exponentially after reaching an initial maximum without an initial steady state. Back-extrapolations to the time of taking the biopsies on the basis of a two-phase exponential decay curve gave baseline thermal power values at 37°C of 41 (D) and 15 (B) μW/mg_dw, respectively, which roughly correspond to known values of basal oxygen consumption of heart and kidney. The higher values of the duodenum may be founded on a higher transport activity in vivo. Altogether, microcalorimetry seems to be a method for comparative evaluation of metabolic activity of human mucosa samples.     

Proteomic analysis of pancreatic ductal carcinoma cells treated with 5-aza-2'-deoxycytidine

A pancreatic adenocarcinoma cell line (PaCa44), which contains, among other alterations, a methylated p16 promoter, was treated with a chemoterapeutic agent, 5-aza-2'-deoxycytidine (DAC), in order to evaluate the effect of this drug on cell growth and protein expression. Cell proliferation was strongly inhibited by a 24 h DAC treatment and this inhibition lasted for at least 10 days. Master maps of control and treated PaCa44 cells were generated by analysis with the PDQuest software. The comparison between such maps showed up- and downregulation of 45 polypeptide chains, of which 32 were downregulated and 13 upregulated, out of a total of 700 spots detected by a medium-sensitivity stain, micellar Coomassie Brilliant Blue. Fingerprinting by mass spectrometry analysis enabled the identification of 36 of these spots. Among the major changes in DAC-treated cells: cofilin and profilin 1 are silenced; coactosin, peptidyl-propyl cis-trans isomerase A and cystatin B are decreased by 22, 16- and 15-fold, respectively; stress-70 protein, superoxide dismutase and protein disulfide isomerase A3 are increased by 13-, 11-, and 5-fold, respectively. The significance of some of these major changes is discussed.