Accurate estimate of pancreatic T2* values: how to deal with fat infiltration

Purpose

We examined different approaches aimed to deal with the signal fluctuation of pancreatic T2* values due to fat infiltration in order to obtain accurate estimates of iron overload.

Methods

Pancreatic T2* values were assessed in 20 patients (13 females, 37.24 ± 9.12 years) enrolled in the Myocardial Iron Overload in Thalassemia network without and with the application of fat suppression-FS (T2*-NoFS and T2*-FS). T2* values were assessed in three different ways: (1) from the immediate fit (original T2*); (2) discarding the echoes until the achievement of a good visual concordance between the signal and the model (final_vis T2*); (3) eliminating the echoes until the achievement of a fitting error (known) <5% (final_thres T2*).

Results

For the T2*-NoFS sequence the original T2* values were significantly higher than the final_vis T2* values (difference:4.8 ± 6.1 ms; P < 0.0001) and the final_thres T2* values (difference:4.3 ± 6.1 ms; P = 0.006). For the T2*-FS sequence the original T2* values were comparable to final_vis and final_thres T2* values. The original T2*-FS values were significantly different from the original T2*-NoFS values. The final_vis T2*-FS values were comparable to the final_vis T2*-NoFS values and the final_thresh T2*-FS values were comparable to the final_thresh T2*-NoFS values. For both T2*-FS and T2*-NoFS sequences, the final_thres T2* values were not significantly different from the final_vis T2* values and no bias was present.

Conclusions

In the clinical practice, an accurate pancreatic iron overload assessment should be done by applying FS and, when needed, by discarding the TEs until the fitting error goes below 5%.

     

Rutin-modified silver nanoparticles as a chromogenic probe for the selective detection of Fe3+ in aqueous medium

The development of nanoprobes for selective detection of metal ions in solution has attracted great attention due to their impact on living organisms. As a contribution to this field, this paper reports the synthesis of silver nanoparticles modified with rutin in the presence of ascorbic acid and their successful use as a chromogenic probe for the selective detection of Fe³⁺ in aqueous solution. Limits of detection and quantification were found to be 17 nmol L⁻¹ and 56 nmol L⁻¹, respectively. The sensing ability is proposed to proceed via an iron-induced nanoparticle growth/aggregation mechanism. A practical approach using image analysis for quantification of Fe³⁺ is also described.

The use of rutin-modified silver nanoparticles for selective detection and sensitive quantification of Fe³⁺ in aqueous solution is described.

Metal ions are key species in nature due to their essential functions in living organisms.^1,2 On the other hand, heavy metals as well as essential metals at abnormally high levels are toxic.² Iron, for instance, in addition to its popular use in industry and construction, is essential to the human body and active in biological processes. Although the trivalent form of iron is particularly important for oxygen transport in blood and the mitochondrial respiratory chain, high levels of this cation are associated with important pathologies.^3,4 The detection of metal ions in aqueous solution is traditionally performed by methods including atomic absorption spectrometry,⁵ electrochemical measurements,^6,7 and inductively coupled plasma techniques,⁸ among others. However, these techniques have important drawbacks, notably the need for sophisticated instrumentation, in addition to being time-consuming and requiring laborious procedures. To overcome these issues, the development of chromogenic and fluorogenic chemosensors for the selective detection of metal-targets has attracted great attention, especially due to the possibility of fast, sensitive and non-expensive analysis.^9,10 In the last decade, nanoscaled materials have been reported as selective probes for metal ions, including Fe³⁺.^11–24Silver nanoparticles (AgNPs) are of particular interest because of the affordable price of starting materials, ease of controlling size and morphology, possibility to functionalize their surface with organic molecules, and optical properties that enable detection of a variety of analytes via simple UV-vis spectroscopy and digital image analysis. Furthermore, applications of AgNPs are also biotechnologically relevant due to the possibility of green synthetic protocols, including the use of plant extracts,²⁵ natural sources,²⁶ glycerol,²⁷ among others.Flavonoids are secondary metabolites naturally found in fruits and other vegetables with relevant roles due to their nutritional, pharmaceutical and medicinal properties.²⁸ Because of their adequate structural features, flavonoids are candidates to be employed in the synthesis of AgNPs.²⁶ Rutin (RU), a sugar-based flavonoid, may be employed as reducing agent in the synthesis of AgNPs along with a stabilizer such as polyvinylpyrrolidone (PVP)²⁶ or used as crude plant extract component.^29,30This paper reports the use of RU-modified AgNPs (RU-AgNPs) as a chromogenic probe for Fe³⁺ in aqueous medium in the presence of ascorbic acid (AA). Sensing ability of RU-AgNPs for the selective detection of Fe³⁺ toward other metal cations was investigated with UV-vis spectroscopy analysis. These data and transmission electronic microscopy (TEM) results allowed a mechanistic proposal involved in the selective detection of Fe³⁺ by RU-AgNPs. Furthermore, a practical approach based on correlation of images of solutions obtained with a conventional smartphone and chemometrics was employed for a simpler quantification of Fe³⁺ in aqueous medium.Initially, the order of reagent combination was investigated in the synthesis of RU-AgNPs. Concentration of RU ranged from 0.10 to 0.50 mmol L⁻¹, while concentrations of other components were fixed at 0.20 mmol L⁻¹ AgNO₃, 0.10 mmol L⁻¹ AA and 0.10 M NaOH. Water was used as solvent in all cases. Narrower surface plasmon resonance (SPR) bands were obtained from adding a solution of AA and NaOH to a solution containing RU and AgNO₃ (Fig. 1a) against the addition of RU, AA, and NaOH to AgNO₃ solution (Fig. S1a – ESI^†), or the addition of RU and NaOH to a solution of AA and AgNO₃ (Fig. S1b^†). RU-AgNPs obtained from the condition presented in Fig. 1a are small (4.1 nm average diameter) and considerably polydisperse (standard deviation of 4.7 nm), however, presenting only one population (Fig. 1b and S2^†). A study of the influence of RU concentration (0.10 to 0.50 mmol L⁻¹) on the stability of RU-AgNPs over time indicated that 0.10 mmol L⁻¹ RU generates more stable RU-AgNPs (Fig. S3^†). Next, a study on the influence of pH indicated that RU-AgNPs are only stable under strong alkaline conditions (pH 12.5 or higher) (Fig. S4^†).Open in a separate windowFig. 1(a) UV-vis analysis of RU-AgNPs under strong alkaline condition (pH > 12.5) based on the order of adding reagents (RU, 0.10 to 0.50 mmol L⁻¹; AgNO₃, 0.20 mmol L⁻¹; AA, 0.10 mmol L⁻¹; NaOH, 0.10 mmol L⁻¹); (b) TEM image of RU-AgNPs under the selected condition.The ability of RU-AgNPs to sense metal cations was investigated by both naked-eye and UV-vis spectroscopy analysis (Fig. 2). The separate addition of 10 μmol L⁻¹ of several metal cations (Fe³⁺, Co²⁺, Zn²⁺, Sr²⁺, Cu²⁺. Al³⁺, Ba²⁺, Cd²⁺, Pb²⁺, Ni²⁺, Mg²⁺, Hg²⁺, Cu⁺ and Cr³⁺) to solutions of RU-AgNPs (prepared according to Fig. 1a) indicated that only Fe³⁺ induces a significant colorimetric change in the final aspect of solution after 50 minutes (Fig. 2a).Open in a separate windowFig. 2Naked-eye (a) and UV-vis (b) analysis of RU-AgNPs in absence (control, C) and presence of 10 μmol L⁻¹ of selected metal cations (Fe³⁺, Co²⁺, Zn²⁺, Sr²⁺, Cu²⁺. Al³⁺, Ba²⁺, Cd²⁺, Pb²⁺, Ni²⁺, Mg²⁺, Hg²⁺, Cu⁺ and Cr³⁺) after 50 min; (c) calibration curve obtained by the addition of different amounts (0 to 10 μmol L⁻¹) of Fe³⁺ to solutions of RU-AgNPs; (d) TEM image of RU-AgNPs after addition of Fe³⁺ (10 μmol L⁻¹). In all experiments, RU-AgNPs were prepared in the presence of ascorbic acid.The results presented in Fig. 2a are consistent with the UV-vis spectroscopy analysis (Fig. 2b). Co²⁺ ions also induce some change in the system, however at a considerably smaller extension than Fe³⁺. In this study, AA played a crucial role in the selective detection of Fe³⁺ by the referred nanoprobe. In the absence of AA, Co²⁺ (mainly) as well as other cations induce stronger colorimetric and spectral changes (Fig. S5a and b,^† respectively) in the analysis of solutions of RU-AgNPs when compared to the system containing AA. This selectivity may arise from two possible reasons: (i) preservation of RU by avoiding its oxidation in the reduction of Ag⁺ ions; (ii) coordination of ascorbate anion to cations other than Fe³⁺.Interaction of RU-AgNPs and Fe³⁺ (10 μmol L⁻¹) stabilizes after approximately 40 minutes (Fig. S6^†). Next, a calibration curve was built from the direct relationship between the absorbance at 396 nm and the concentration of Fe³⁺ (Fig. 2c), presenting a good correlation (R² = 0.9929). The limits of detection and quantification were found to be 17 nmol L⁻¹ and 56 nmol L⁻¹, respectively, which is very satisfactory.^13,14 The influence of other cations in the detection of Fe³⁺ was investigated by UV-vis spectroscopy. Fig. S7^† clearly demonstrates that there are only small changes when a second cation (30 μmol L⁻¹) is added together with Fe³⁺ to the RU-AgNPs solution.Mechanistically, the detection of Fe³⁺ by RU-AgNPs in aqueous medium proceeds via a growth/aggregation-combined process. This proposal is first evidenced by UV-vis analysis due the suppression of SPR band (Fig. 2b), a behavior consistent with the literature.^31,32 Interestingly, TEM analysis clearly shows a growth in AgNPs size after addition of Fe³⁺ to the solution (Fig. 2d), resulting in a final single AgNPs population with average diameter of 14.7 nm ± 8.9 nm. Due to the strong alkaline medium, the main specie responsible for the behavior of NPs is likely to be Fe(OH)₃. A schematic illustration of the mechanism involving aggregation of RU-AgNPs induced by the addition of Fe³⁺ is presented in Fig. 3. AgNPs are initially formed by adsorption of anionic RU to the silver surface via deprotonated 5-hydroxychromen-4-one moiety. This is supported by the literature³³ and confirmed by alteration in the 1800–1500 cm⁻¹ region of the RU infrared spectra before and after coordination with silver (Fig. S9^†). Afterwards, the addition of Fe³⁺ induces the formation of a coordination complex through an anionic catechol group, in which at least 2 : 1 ligand–Fe³⁺ stoichiometry is required for an aggregated effect.Open in a separate windowFig. 3Mechanistic proposal for growth/aggregation of RU-AgNPs in the presence of Fe³⁺. Insert: binding model for RU-AgNPs.Due to increasing interest in image processing as an analytical tool for many purposes,^34,35 Multiple Linear Regression was employed to verify the capacity of the RU-AgNPs to probe Fe³⁺ standards at distinct concentrations, as presented in Fig. 4. The curve was obtained by plotting the color absorbances RGB-based values versus the concentrations of Fe³⁺ standards after RU-AgNPs interaction. Predicted iron is a vector based on RGB values that were then extracted from the filtered images and inserted in the equation described by Beer–Lambert law in order to generate the absorbances for the construction of the analytical curve. A linear behavior between the predicted response and the measured concentrations was observed (R² = 0.9806).Open in a separate windowFig. 4Calibration curve for Fe³⁺ analysis showing the predicted iron (RGB) vs. iron(iii) concentration (1 to 8 μmol L⁻¹). Adjusted R² = 0.9806.Regression coefficients of the calibration model (using the R, G and B channels simultaneously) obtained by MLR method are shown in eqn (1):[Fe³⁺] = 44.5R − 4.9G − 16.4B + 5.11where Fe³⁺ concentration is the dependent variable (Predicted Iron), 5.1 is the intercept (β₀), 44.5, −4.9 and −16.4 are the regression coefficients of the independent variables (R, G and B channels, respectively).It is possible to observe an interesting performance of the method using the three RGB color channels allied with MLR to quantify the Fe³⁺ content, which presented reasonable deviations in its responses considering that it is simple and low-cost. The good linearity is similar to other colorimetric methods such as sodium determination in seawater and coconut water (R² > 0.91) by Moraes and coauthors,³⁶ or iron(ii) in simulated seawater (R² = 0.9993) by Gasparotto et al.³⁷Second order regression was applied to the dataset to obtain a better adjustment, resulting in an adjusted R-squared of 0.9955. RGB values were then inserted in eqn (2) in order to generate the construction of the correlation:[Fe³⁺] = 14.4R + 30.7G − 23.4B − 398RG − 348RB − 0.8BG − 978R² + 695G² + 26.9B² + 4.52This paper reports the use of AgNPs functionalized with RU as nanoprobes for selective detection and sensitive quantification of Fe³⁺ in aqueous solution. The synthesis of RU-AgNPs is reproducible, easily performed and requires no stabilizer agent other than RU. AA has a crucial role in the selectivity by either the avoidance of oxidation of RU by silver and/or coordination of ascorbate with other cations. The literature brings relevant examples of chromogenic and fluorogenic chemosensors for selective detection of Fe³⁺ in solution. Many of these artificial organic receptors present high selectivity and relevant limits of detection, requiring, however, very specific reagents and laborious synthetic procedures.^38–41 Metal-based nanoparticles have emerged as potential probes for detection of Fe³⁺.^11–16 Although effective in Fe³⁺ sensing, the synthesis of these nanoprobes require some toxic reagents, such as NaBH₄ or PVP, use plant extracts, which may lead to some drawbacks, such as the understanding of the sensing mechanism. In contrast, our method is based on commercially available, nontoxic, low-cost reagents. Fe³⁺ sensing performed satisfactorily in the 1–10 μmol L⁻¹ range, and the limit of detection obtained with this method (17 nmol L⁻¹) is comparable to the most sensitive methods reported in literature. A mechanism for the detection of Fe³⁺ by RU-AgNPs involves a combined growth/aggregation of the NPs. There is a still limited number of nanoscaled systems reported as being selective and sensitive in the detection of Fe³⁺, which reinforces the relevance of the method reported herein. The linearity range obtained by both UV-vis spectroscopy and image analysis comprises the maximum of residual Fe³⁺ in drinking water according to the European and US legislations.^15,42     

Molecular events underlying interleukin‐6 independence in a subclone of the CMA‐03 multiple myeloma cell line

We explored the molecular mechanisms involved in the establishement of CMA‐03/06, an IL‐6‐independent variant of the multiple myeloma cell line CMA‐03 previously generated in our Institution. CMA‐03/06 cells grow in the absence of IL‐6 with a doubling time comparable with that of CMA‐03 cells; neither the addition of IL6 (IL‐6) to the culture medium nor co‐culture with multipotent mesenchymal stromal cells increases the proliferation rate, although they maintain the responsiveness to IL‐6 stimulation as demonstrated by STAT1, STAT3, and STAT5 induction. IL‐6 independence of CMA‐03/06 cells is not apparently due to the development of an autocrine IL‐6 loop, nor to the observed moderate constitutive activation of STAT5 and STAT3, since STAT3 silencing does not affect cell viability or proliferation. When compared to the parental cell line, CMA‐03/06 cells showed an activated pattern of the NF‐κB pathway. This finding is supported by gene expression profiling (GEP) analysis identifying an appreciable fraction of modulated genes (28/308) in the CMA‐03/06 subclone reported to be involved in this pathway. Furthermore, although more resistant to apoptotic stimuli compared to the parental cell line, CMA‐03/06 cells display a higher sensibility to NF‐κB inhibition induced by bortezomib. Finally, GEP analysis suggests an involvement of a number of cytokines, which might contribute to IL‐6 independence of CMA‐03/06 by stimulating growth and antiapoptotic processes. In conclusion, the parental cell‐line CMA‐03 and its variant CMA‐03/06 represent a suitable model to further investigate molecular mechanisms involved in the IL‐6‐independent growth of myeloma cells. © 2013 Wiley Periodicals, Inc.     

Delaying aging and the aging-associated decline in protein homeostasis by inhibition of tryptophan degradation

Toxicity of aggregation-prone proteins is thought to play an important role in aging and age-related neurological diseases like Parkinson and Alzheimer's diseases. Here, we identify tryptophan 2,3-dioxygenase (tdo-2), the first enzyme in the kynurenine pathway of tryptophan degradation, as a metabolic regulator of age-related α-synuclein toxicity in a Caenorhabditis elegans model. Depletion of tdo-2 also suppresses toxicity of other heterologous aggregation-prone proteins, including amyloid-β and polyglutamine proteins, and endogenous metastable proteins that are sensors of normal protein homeostasis. This finding suggests that tdo-2 functions as a general regulator of protein homeostasis. Analysis of metabolite levels in C. elegans strains with mutations in enzymes that act downstream of tdo-2 indicates that this suppression of toxicity is independent of downstream metabolites in the kynurenine pathway. Depletion of tdo-2 increases tryptophan levels, and feeding worms with extra l-tryptophan also suppresses toxicity, suggesting that tdo-2 regulates proteotoxicity through tryptophan. Depletion of tdo-2 extends lifespan in these worms. Together, these results implicate tdo-2 as a metabolic switch of age-related protein homeostasis and lifespan. With TDO and Indoleamine 2,3-dioxygenase as evolutionarily conserved human orthologs of TDO-2, intervening with tryptophan metabolism may offer avenues to reducing proteotoxicity in aging and age-related diseases.     

Psychosocial correlates of sleep quality and architecture in women with metastatic breast cancer

Background

Sleep disturbance is prevalent among women with metastatic breast cancer (MBC). Our study examined the relationship of depression and marital status to sleep assessed over three nights of polysomnography (PSG).

Methods

Women with MBC (N = 103) were recruited; they were predominately white (88.2%) and 57.8 ± 7.7 years of age. Linear regression analyses assessed relationships among depression, marital status, and sleep parameters.

Results

Women with MBC who reported more depressive symptoms had lighter sleep (e.g., stage 1 sleep; P < .05), less slow-wave sleep (SWS) (P < .05), and less rapid eye movement (REM) sleep (P < .05). Single women had less total sleep time (TST) (P < .01), more wake after sleep onset (WASO) (P < .05), worse sleep efficiency (SE) (P < .05), lighter sleep (e.g., stage 1; P < .05), and less REM sleep (P < .05) than married women. Significant interactions indicated that depressed and single women had worse sleep quality than partnered women or those who were not depressed.

Conclusion

Women with MBC and greater symptoms of depression had increased light sleep and reduced SWS and REM sleep, and single women had worse sleep quality and greater light sleep than married counterparts. Marriage was related to improved sleep for women with more depressive symptoms.     

Hepatitis C virus infection and health-related quality of life

Hepatitis C virus (HCV) hepatitis and other diseases related to HCV, such as cryoglobulinemia, lymphoma and renal failure, impair health-related quality of life (HRQoL). In addition, HCV per se might directly influence HRQoL via colonization of microglia in the brain or, indirectly, via the effect of systemic inflammatory cytokines which, in turn, can trigger brain interleukin production. The treatment of HCV-related disorders with interferon (IFN) has an effect on HRQoL. Initially, IFN causes a transient deterioration of HRQoL, due to the induction of depression and other side effects of treatment. Subsequently, the subjects who obtain a sustained virologic response experience an improvement in HRQoL. Only rarely does interferon treatment causes permanent detrimental effects on HRQoL, due to residual psychiatric or neurologic side effects. Liver transplantation is the only treatment for end-stage HCV-related liver disease. HRQoL generally improves massively a few months after transplantation, except in the case of serious complications of the transplant procedure. Furthermore, high levels of anxiety and neuroticism pre-transplant are associated with lower HRQoL one year after transplant. Additionally, six months after transplant, patients with HCV who experience virologic recurrence show significantly greater depression, anxiety, phobic anxiety, and paranoid ideation than anti-HCV-negative patients. In conclusion, optimal care for the overall well-being of patients with HCV infection requires adequate knowledge of their neurological and psychological status.     

New pharmacotherapy options for noninfectious posterior uveitis

International Ophthalmology - Noninfectious inflammation of the posterior eye segment represents an important cause of visual impairment. It often affects relatively young people and causes a...