Probing metabolic alterations in breast cancer in response to molecular inhibitors with Raman spectroscopy and validated with mass spectrometry

Rapid and accurate response to targeted therapies is critical to differentiate tumors that are resistant to treatment early in the regimen. In this work, we demonstrate a rapid, noninvasive, and label-free approach to evaluate treatment response to molecular inhibitors in breast cancer (BC) cells with Raman spectroscopy (RS). Metabolic reprogramming in BC was probed with RS and multivariate analysis was applied to classify the cells into responsive or nonresponsive groups as a function of drug dosage, drug type, and cell type. Metabolites identified with RS were then validated with mass spectrometry (MS). We treated triple-negative BC cells with Trametinib, an inhibitor of the extracellular-signal-regulated kinase (ERK) pathway. Changes measured with both RS and MS corresponding to membrane phospholipids, amino acids, lipids and fatty acids indicated that these BC cells were responsive to treatment. Comparatively, minimal metabolic changes were observed post-treatment with Alpelisib, an inhibitor of the mammalian target of rapamycin (mTOR) pathway, indicating treatment resistance. These findings were corroborated with cell viability assay and immunoblotting. We also showed estrogen receptor-positive MCF-7 cells were nonresponsive to Trametinib with minimal metabolic and viability changes. Our findings support that oncometabolites identified with RS will ultimately enable rapid drug screening in patients ensuring patients receive the most effective treatment at the earliest time point.

Rapid and accurate response to targeted therapies is critical to differentiate tumors that are resistant to treatment early in the regimen.     

FTIR Spectroscopy as a Tool to Study Age-Related Changes in Cardiac and Skeletal Muscle of Female C57BL/6J Mice

Studying aging is important to further understand the molecular mechanisms underlying this physiological process and, ideally, to identify a panel of aging biomarkers. Animals, in particular mice, are often used in aging studies, since they mimic important features of human aging, age quickly, and are easy to manipulate. The present work describes the use of Fourier Transform Infrared (FTIR) spectroscopy to identify an age-related spectroscopic profile of the cardiac and skeletal muscle tissues of C57BL/6J female mice. We acquired ATR-FTIR spectra of cardiac and skeletal muscle at four different ages: 6; 12; 17 and 24 months (10 samples at each age) and analyzed the data using multivariate statistical tools (PCA and PLS) and peak intensity analyses. The results suggest deep changes in protein secondary structure in 24-month-old mice compared to both tissues in 6-month-old mice. Oligomeric structures decreased with age in both tissues, while intermolecular β-sheet structures increased with aging in cardiac muscle but not in skeletal muscle. Despite FTIR spectroscopy being unable to identify the proteins responsible for these conformational changes, this study gives insights into the potential of FTIR to monitor the aging process and identify an age-specific spectroscopic signature.     

High throughput screening technology and the small molecules modulating aging related signals

Aging and its related diseases are severe issues in modern society. Many efforts have been made to understand the mechanisms of aging and to find the ways to prevent age-related diseases. Identifying the compounds targeting aging-related signals is a challenging work because there are so many proteins and signals involved. Recently, alone with the progresses in high throughput screening (HTS) technology, increasing numbers of small molecules targeting aging-related pathologic processes have been identified. In this review, we introduce the basic workflow, classification and assay strategies of HTS technology, and sort out known small molecules identified via HTS technology by their roles in aging related diseases, such as neural degenerative diseases, diabetes and tumors. Given the fact that application of HTS on aging research is still at an early stage, we also summarize the cellular mechanisms about aging process, paralleled with the compounds which can modulate the functions of proteins important for aging signals. Finally, we briefly discuss some advanced HTS technologies for their potent applications on the discovery of anti-aging compounds. The main purpose of this review is to provide updated and useful information to those who are interested in pharmacology and HTS technology, but not familiar with aging biology, or vice versa.     

Metabolomic analysis of amino acids and organic acids in aging mouse eyes using gas chromatography–tandem mass spectrometry

This is a metabolomics study for monitoring altered amino acid (AA) and organic acid (OA) metabolism of in eyes from aging an mouse model at 8 and 18 weeks and 18 months. Simultaneous metabolic profiling analysis of OAs and AAs was performed as ethoxycarbonyl/methoxime/tert-butyldimethylsilyl derivatives by gas chromatography–tandem mass spectrometry. A total of 42 metabolites—24 AAs and 18 OAs—were determined and their composition values were normalized to the corresponding mean values of 8-week-old mice as the control group. Then their normalized values were plotted as star graphs, which were distorted and readily distinguishable for each age-related group. Among the 42 metabolites, 18 AAs and 11 OAs were age dependent and significantly different (p < 0.05). Principal component analysis and partial least squares discriminant analysis showed unclear separation between 8- and 18-week-old mice but clear separation between these and 18-month-old mice. In particular, the variable importance in projection scores of 4-hydroxyproline, cis-aconitic acid, glycine, isocitric acid, leucine, pipecolic acid and lysine from partial least-squares–discriminant analysis were higher than 1.3. A heatmap for the classification and visualization of 42 metabolites showed differences in metabolite changes with aging. Altered AA and OA profiles were monitored, which may explain the metabolic disturbance of AA and OA. These findings are related to mitochondrial dysfunctions related to energy metabolism and the impaired antioxidant system in the aging eye. Therefore, the present metabolomics results of the association between physiological states and altered metabolism of AA and OA will be useful for understanding the aging eye and related diseases.     

Morphology,dynamics, and order development in a thermoplastic polyurethane with melt blended POSS

A top‐down approach is applied for the production of polyurethane (PU)–polyhedral oligomeric silsesquioxane (POSS) nanocomposites, namely melt blending. As opposed to the typical chemical incorporation during synthesis, a POSS moiety with two hydroxyl groups is melt blended into a commercial thermoplastic polyurethane with mass fraction up to 2 wt %. POSS disperses in the matrix in submicrometer‐sized crystals, as well as in length scale of few tens of nanometers, in the bulk. Phase separation of the produced composites was studied by both standard dynamic and isothermal annealing experiments. In an approach rare in the literature, the dynamics of phase separation is discussed based on isothermal differential scanning calorimetry curves recorded during annealing. The blended‐in nanoparticles affect the micromorphology in a complicated manner, dependent on the intrinsically complex phase separation mechanism of PU. At higher temperatures, POSS slows down the phase separation, whereas at lower ones, it enhances and accelerates it. POSS decreases the mechanical modulus of the final material, presumably as a result of changes in the microphase separation. © 2019 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2019 , 57, 1133–1142     

Ex vivo evaluation of the effects of a white grape juice extract on lymphocytic mitochondrial functions

Abstract

The physio-pathological role of mitochondria in aging and age-related diseases has stimulated the search for compounds able to promote mitochondrial functionality. Our study was designed to evaluate the effect of a white grape juice extract (WGJe) on mitochondrial activity, in an ex vivo experimental model consisting of activated lymphocytes obtained from a younger age group and an older age group of subjects. WGJe steadily decreased the lymphocytic mitochondrial mass in the older subjects, without a relevant effect in their younger counterpart, and significantly enhanced Δψm in both groups investigated. Finally, WGJe reduced the endogenous mitochondrial production of H₂O₂ in all subjects. The results support the potential use of WGJe to improve mitochondrial functionality, thus maintaining human health and slowing down aging.     

High-resolution time-of-flight mass spectrometry fingerprinting of metabolites from cecum and distal colon contents of rats fed resistant starch

Time-of-flight mass spectrometry along with statistical analysis was utilized to study metabolic profiles among rats fed resistant starch (RS) diets. Fischer 344 rats were fed four starch diets consisting of 55 % (w/w, dbs) starch. A control starch diet consisting of corn starch was compared against three RS diets. The RS diets were high-amylose corn starch (HA7), HA7 chemically modified with octenyl succinic anhydride, and stearic-acid-complexed HA7 starch. A subgroup received antibiotic treatment to determine if perturbations in the gut microbiome were long lasting. A second subgroup was treated with azoxymethane (AOM), a carcinogen. At the end of the 8-week study, cecal and distal colon content samples were collected from the sacrificed rats. Metabolites were extracted from cecal and distal colon samples into acetonitrile. The extracts were then analyzed on an accurate-mass time-of-flight mass spectrometer to obtain their metabolic profile. The data were analyzed using partial least-squares discriminant analysis (PLS-DA). The PLS-DA analysis utilized a training set and verification set to classify samples within diet and treatment groups. PLS-DA could reliably differentiate the diet treatments for both cecal and distal colon samples. The PLS-DA analyses of the antibiotic and no antibiotic-treated subgroups were well classified for cecal samples and modestly separated for distal colon samples. PLS-DA analysis had limited success separating distal colon samples for rats given AOM from those not treated; the cecal samples from AOM had very poor classification. Mass spectrometry profiling coupled with PLS-DA can readily classify metabolite differences among rats given RS diets.     

Curcumin as Prospective Anti-Aging Natural Compound: Focus on Brain

The nutrients and their potential benefits are a new field of study in modern medicine for their positive impact on health. Curcumin, the yellow polyphenolic compound extracted from Curcuma longa species, is widely used in traditional Ayurvedic medicine to prevent and contrast many diseases, considering its antioxidant, immunomodulatory, anti-inflammatory, anti-microbial, cardio-protective, nephron-protective, hepato-protective, anti-neoplastic, and anti-rheumatic proprieties. In recent years, the investigations of curcumin have been focused on its application to aging and age-associated diseases. Aging is a physiological process in which there is a decreasing of cellular function due to internal or external stimuli. Oxidative stress is one of the most important causes of aging and age-related diseases. Moreover, many age-related disorders such as cancer, neuroinflammation, and infections are due to a low-grade chronic systemic inflammation. Curcumin acting on different proteins is able to contrast both oxidative stress than inflammation. In the brain, curcumin is able to modulate inflammation induced by microglia. Finally in brain tumors curcumin is able to reduce tumor growth by inhibition of telomerase activity. This review emphasizes the anti-aging role of curcumin focusing on its mechanism to counteract aging in the brain. Moreover, new formulations to increase the bioavailability of curcumin are discussed.     

Stability and aging of solubilized dialdehyde cellulose

Derivatization of alpha cellulose by periodate oxidation is an useful method for production of dialdehyde cellulose (DAC). Conversion of the 2,3-hydroxyl groups to a pair of aldehyde groups along with cleavage of the C2–C3 bond of anhydroglucose unit reduces crystallinity of initial material, leaving DAC soluble in water under mild conditions. Solubilization in hot water is necessary to obtain product in solution. The first part of our work confirmed that solubilization causes severe degradation of the molecular weight of the polymer. However, the chemistry and processes within these solutions are currently poorly understood. In the main part of the study, products of periodate oxidation were identified in acidic DAC solutions by NMR spectroscopy for the first time. Subsequent investigation of the acidic DAC solution's aging demonstrated that the low pH of the DAC solution considerably slows the degradation processes, namely the decrease of reactive aldehyde group content when compared to previous studies. Large increase in the molecular weight, observed after 14 days of aging, was explained by formation of intermolecular hemiacetals. Our results demonstrate that pH-stabilized aqueous DAC solutions remained active (e.g. applicable for cross-linking reactions) even several weeks after preparation, therefore reducing the need to prepare a fresh solution each time.     

人类钙营养状况,缺钙后果及其对策

人类缺钙是世界性普遍存在的问题，中国人由于膳食结构缺陷及传统饮食习惯而使食物中钙的吸收受到影响，因而缺钙尤其普遍。     

Inflammaging and Brain: Curcumin and Its Beneficial Potential as Regulator of Microglia Activation

Inflammaging is a term used to describe the tight relationship between low-grade chronic inflammation and aging that occurs during physiological aging in the absence of evident infection. This condition has been linked to a broad spectrum of age-related disorders in various organs including the brain. Inflammaging represents a highly significant risk factor for the development and progression of age-related conditions, including neurodegenerative diseases which are characterized by the progressive dysfunction and degeneration of neurons in the brain and peripheral nervous system. Curcumin is a widely studied polyphenol isolated from Curcuma longa with a variety of pharmacologic properties. It is well-known for its healing properties and has been extensively used in Asian medicine to treat a variety of illness conditions. The number of studies that suggest beneficial effects of curcumin on brain pathologies and age-related diseases is increasing. Curcumin is able to inhibit the formation of reactive-oxygen species and other pro-inflammatory mediators that are believed to play a pivotal role in many age-related diseases. Curcumin has been recently proposed as a potential useful remedy against neurodegenerative disorders and brain ageing. In light of this, our current review aims to discuss the potential positive effects of Curcumin on the possibility to control inflammaging emphasizing the possible modulation of inflammaging processes in neurodegenerative diseases.     

Thermo-oxidative aging in surface layer of cis-1,4-polybutadiene vulcanizates

A study of the microstructural changes occurring in the surface layer of cis-1,4-polybutadiene (BR) vulcanizates during thermo-oxidative aging in the temperature range of 120–180°C was carried out by using optical microscope, SEM, MIR–IR, XPS, and swelling. The physical properties, structural changes, and interreaction between rubber and filler (clay) in the surface layer comparing with the center bulk of BR vulcanizates are observed.     

Synthesis and Biological Evaluation of Halogenated E-Stilbenols as Promising Antiaging Agents

The increased risk of illness and disability is related to the age inevitable biological changes. Oxidative stress is a proposed mechanism for many age-related diseases. The crucial importance of polyphenol pharmacophore for aging process is largely described thanks to its effects on concentrations of reactive oxygen species. Resveratrol (3,5,4′-trihydroxy-trans-stilbene, RSV) plays a critical role in slowing the aging process but has a poor bioavailabity after oral intake. In this present work, a series of RSV derivatives was designed, synthesized, and evaluated as potential antioxidant agents. These derivatives contain substituents with different electronic and steric properties in different positions of aromatic rings. This kind of substituents affects the activity and the bioavailability of these compounds compared with RSV used as reference compound. Studies of Log P values demonstrated that the introduction of halogens gives the optimum lipophilicity to be considered promising active agents. Among them, compound 6 showed the higher antioxidant activity than RSV. The presence of trifluoromethyl group together with a chlorine atom increased the antioxidant activity compared to RSV.     

PHOTOSENSITIZED OXIDATION IN THE OCULAR LENS: EVIDENCE FOR PHOTOSENSITIZERS ENDOGENOUS TO THE HUMAN LENS

Abstract— Numerous investigators have attempted to associate near UV light exposure with various changes which occur to lens crystallins during aging and cataractogenesis. Recently we have shown in vitro that singlet oxygen mediated oxidation of lens crystallins produces effects very similar to those documented for crystallins from old or cataractous lenses and suggested that near UV photodynamic effects may play a major role in vivo in aging in the human lens. In the present work we demonstrate that certain oxidation products of tryptophan which have been identified in human lens can act as near UV photosensitizers, producing singlet oxygen. The insoluble protein fraction from human cataracts is shown to have the capacity to act as a photosensitizer. An age-related increase in photosensitizing capacity is also demonstrated in the soluble crystallins from human lens. These findings are discussed with respect to development of pigmented nuclear cataracts.     

Role of Melatonin in Angiotensin and Aging

The cellular utilization of oxygen leads to the generation of free radicals in organisms. The accumulation of these free radicals contributes significantly to aging and several age-related diseases. Angiotensin II can contribute to DNA damage through oxidative stress by activating the NAD(P)H oxidase pathway, which in turn results in the production of reactive oxygen species. This radical oxygen-containing molecule has been linked to aging and several age-related disorders, including renal damage. Considering the role of angiotensin in aging, melatonin might relieve angiotensin-II-induced stress by enhancing the mitochondrial calcium uptake 1 pathway, which is crucial in preventing the mitochondrial calcium overload that may trigger increased production of reactive oxygen species and oxidative stress. This review highlights the role and importance of melatonin together with angiotensin in aging and age-related diseases.     

Changes in curing behaviour of aminoresins during storage

Summary The curing behaviour of commercial UF and MUF resins, stored at room temperature nearly up to gelation, is studied by simultaneous TG-DTA technique and structural changes of resins are also followed during aging. On the basis of ¹³C NMR spectra, the main chemical reaction during UF resin storage is the formation of methylenes and dimethylene ethers linked to secondary amino groups. Aging of resins results in a decrease of cure rate which is related to lower concentration of active functional groups and decrease in molecular mobility. On DTA curve, the resin with higher content of methylol groups reveals the curing exotherm earlier. With decreasing methylol content during storage, the peak maximum of exotherm is shifted to higher temperature value. Advanced polycondensation and sedimentation processes during storage produce partly locked in macromolecule structure water, and the water evaporation endotherm on DTA curve shifts to considerably higher temperature. The aged MUF resins are chemically less changed than UF resins and the aging process mainly involves noncovalent network formation due to complex molecular structure.     

Age-related decline in expression of calnexin

Aging is accompanied by the changes in the cells that decrease their capacity to respond to various forms of stress. Cells are known to respond to stresses through expression of stress-response proteins, heat-shock proteins composed of molecular chaperones. Recent studies suggest that chaperone level and stress-induced chaperone expression could decrease with aging. The aim of the present study is to identify chaperones that show a significant change in protein expression with aging. We used an in vitro aging model system of human diploid fibroblasts (HDF). Proteome analysis of HDF showed that endoplasmic reticulum (ER) chaperone, calnexin, significantly decreased with aging. Oxidative stress-induced expression of calnexin also attenuated in old HDF compared to young cells. These findings suggest calnexin decreases with aging and might contribute to a cytoprotection in a variety of human age-related diseases.     

Physical aging of syndiotactic polypropylene

Syndiotactic polypropylene (sPP) was quenched from the melt in an ice‐water bath, and changes in the structural organization, during the aging time, were followed by X‐rays, differential scanning calorimetry, and transport properties of dichloromethane at low activities. After 1 month, an increase of crystallinity from 19 to 26% was observed. In addition, the results of sorption and diffusion indicated a consistent increase of an intermediate phase not crystalline, yet impermeable to the vapors. The study of the mechanical properties showed that there is a remarkable increase of all the mechanical parameters with the aging time, and this effect was associated to the increase of the intermediate phase. The elastic modulus increased three times in the first hours of aging. © 1999 John Wiley & Sons, Inc. J Polym Sci B: Polym Phys 37: 173–180, 1999     

Phytotherapeutic Approaches to the Prevention of Age-Related Changes and the Extension of Active Longevity

Maintaining quality of life with an increase in life expectancy is considered one of the global problems of our time. This review explores the possibility of using natural plant compounds with antioxidant, anti-inflammatory, anti-glycation, and anti-neurodegenerative properties to slow down the onset of age-related changes. Age-related changes such as a decrease in mental abilities, the development of inflammatory processes, and increased risk of developing type 2 diabetes have a significant impact on maintaining quality of life. Herbal preparations can play an essential role in preventing and treating neurodegenerative diseases that accompany age-related changes, including Alzheimer’s and Parkinson’s diseases. Medicinal plants have known sedative, muscle relaxant, neuroprotective, nootropic, and antiparkinsonian properties. The secondary metabolites, mainly polyphenolic compounds, are valuable substances for the development of new anti-inflammatory and hypoglycemic agents. Understanding how mixtures of plants and their biologically active substances work together to achieve a specific biological effect can help develop targeted drugs to prevent diseases associated with aging and age-related changes. Understanding the mechanisms of the biological activity of plant complexes and mixtures determines the prospects for using metabolomic and biochemical methods to prolong active longevity.