Search

Purpose of review The branched-chain amino acids (BCAA), branched-chain keto acids (BCKA), and β-hydroxy β-methylbutyric acid (HMB) have regained interest as food ingredients in health and disease. To support nutritional strategies, it is critical to gain insight into the whole body and transorgan kinetics of these components. We, therefore, reviewed the most recent literature in this field on in vivo analysis of BCAA, BCKA, and HMB kinetics in health and disease. Recent findings With a new comprehensive metabolic flux analysis BCAA, BCKA, and HMB whole body production, interconversion and disposal rates can be measured simultaneously. Recent studies have provided us with a better understanding of whole-body and transorgan kinetics under postabsorptive, postprandial, hibernating, and lactating conditions. In human pathophysiological conditions like COPD, obesity, and diabetes, the added value of BCAA kinetic measurements over the commonly used concentration measurements only, is discussed. Summary This article highlights the importance of implementing BCAA, BCKA, and HMB kinetic studies to further advance the field by gaining more mechanistic insights and providing direction to the development of new targeted (nutritional) strategies.

Peripheral artery disease (PAD) in the lower limb compromises oxygen supply due to arterial occlusion. Ischemic skeletal muscle is accompanied by capillary structural deformation. Therefore, using novel microscopy techniques, we tested the hypothesis that endothelial cell swelling temporally and quantitatively corresponds to enhanced microvascular permeability. Hindlimb ischemia was created in male Wistar rat’s by iliac artery ligation (AL). The tibialis anterior (TA) muscle microcirculation was imaged using intravenously infused rhodamine B isothiocyanate dextran fluorescent dye via two-photon laser scanning microscopy (TPLSM) and dye extravasation at 3 and 7 days post-AL quantified to assess microvascular permeability. The TA microvascular endothelial ultrastructure was analyzed by transmission electron microscopy (TEM). Compared with control (0.40 ± 0.15 μm3 × 106), using TPLSM, the volumetrically determined interstitial leakage of fluorescent dye measured at 3 (3.0 ± 0.40 μm3 × 106) and 7 (2.5 ± 0.8 μm3 × 106) days was increased (both P < 0.05). Capillary wall thickness was also elevated at 3 (0.21 ± 0.06 μm) and 7 (0.21 ± 0.08 μm) days versus control (0.11 ± 0.03 μm, both P < 0.05). Capillary endothelial cell swelling was temporally and quantitatively associated with elevated vascular permeability in the AL model of PAD but these changes occurred in the absence of elevations in protein levels of vascular endothelial growth factor (VEGF) its receptor (VEGFR2 which decreased by AL-7 day) or matrix metalloproteinase. The temporal coherence of endothelial cell swelling and increased vascular permeability supports a common upstream mediator. TPLSM, in combination with TEM, provides a sensitive and spatially discrete technique to assess the mechanistic bases for, and efficacy of, therapeutic countermeasures to the pernicious sequelae of compromised peripheral arterial function.

Enniatins (Enns) are mycotoxins produced by Fusarium spp. which are a fungus widely spread throughout cereals and cereal-based products. Among all the identified enniatins, Enn B1 stands as one of the most prevalent analogues in cereals in Europe. Hence, the aim of this study was to evaluate for the first time the presence of Enn B1 and its phase I metabolites in 300 human urine samples using an ultrahigh-performance liquid chromatography high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) methodology. Enn B1 was detected in 94.3% of samples ranging from 0.007 to 0.429 ng/mL (mean value: 0.065 ng/mL). In accordance with previous in vitro and in vivo analysis, hydroxylated metabolites (78.0% samples) and carbonylated metabolites (66.0% samples) were tentatively identified as the major products. Results from this biomonitoring study point to a frequent intake of Enn B1 in the studied population, suggesting that in-depth toxicological studies are needed in order to understand the potential effects in humans.

F1-protein fraction (F1) is a natural bioactive compound extracted from the rubber tree, Hevea brasiliensis, and has been recently studied for its therapeutic potential in wound healing. In this study, we investigated the concentration-dependent effects of F1 (0.01%, 0.025%, 0.05%, and 0.1%) incorporated into deproteinized bovine bone (DBB) and porous biphasic calcium phosphate (pBCP), on the repair of rat calvarial critical-size bone defects (CSBD). The defects were analyzed by 3D-microtomography and 2D-histomorphometry at 12 weeks postsurgery. The binding efficiency of F1 to pBCP (96.3 ± 1.4%) was higher than that to DBB (67.7 ± 3.3%). In vivo analysis showed a higher bone volume (BV) gain in all defects treated with DBB (except in 0.1% of F1) and pBCP (except in 0.05% and 0.1% of F1) compared to the CSBD without treatment/control group (9.96 ± 2.8 mm3 ). DBB plus 0.025% F1 promoted the highest BV gain (29.7 ± 2.2 mm3 , p .05). Additionally, a small BV occurred in defects treated with pBCP plus 0.1% F1 (10.4 ± 1.4 mm3, p < .05). In conclusion, F1 showed a higher bone formation potential in combination with DBB than with pBCP, in a concentration-dependent manner. Incorporation of 0.25% F1 into DBB showed the best results with respect to bone formation/repair in CSBD. These results suggest that DBB plus 0.25% F1 can be used as a promising bioactive material for application in bone tissue engineering.

Background Dendrobii Officinalis Caulis (DC) is a well-known tonic herbal medicine worldwide and has favorable immunomodulatory activity. Various material specifications of DC are available in herbal markets, and DC is ingested by different edible methods. However, whether these specifications and edible methods are suitable or not remains unknown. Methods In this study, we evaluated the suitability of four material specifications (fresh stem, dried stem, fengdou and powder) and three edible methods (making tea, soup and medicinal liquor) based on holistic polysaccharide marker (HPM), the major polysaccharide components in DC. First, the HPMs were extracted from the four specifications of DC by the three edible methods in different conditions. Second, qualitative and quantitative characterization of the extracted HPMs was performed using high performance gel permeation chromatography (HPGPC). Third, immunomodulatory activities of the extracted HPMs were evaluated in vivo. Results The results showed that the HPMs were found to be quantitatively different from various specification of DC and edible methods. In vivo analysis indicated that the HPMs exerted positive effects on innate immune responses by increment in proliferation of splenocytes, secretion of IL-2 and cytotoxicity activity of NK cells. Moreover, the dosage amount of HPM should be defined as a certain range, but not the larger the better, for exerting strong immunological activities. Conclusion According to the both chemical and biological results, fengdou by boiling with water for 4 h is the most recommended specification and edible method for DC.

Raman spectroscopy is a non-invasive technique that allows specific chemical information to be obtained from various types of sample. The detailed molecular information that is present in Raman spectra permits monitoring of biochemical changes that occur in diseases, such as cancer, and can be used for the early detection and diagnosis of the disease, for monitoring treatment, and to distinguish between cancerous and non-cancerous biological samples. Several techniques have been developed to enhance the capabilities of Raman spectroscopy by improving detection sensitivity, reducing imaging times and increasing the potential applicability for in vivo analysis. The different Raman techniques each have their own advantages that can accommodate the alternative detection formats, allowing the techniques to be applied in several ways for the detection and diagnosis of cancer. This feature article discusses the various forms of Raman spectroscopy, how they have been applied for cancer detection, and the adaptation of the techniques towards their use for in vivo cancer detection and in clinical diagnostics. Despite the advances in Raman spectroscopy, the clinical application of the technique is still limited and certain challenges must be overcome to enable clinical translation. We provide an outlook on the future of the techniques in this area and what we believe is required to allow the potential of Raman spectroscopy to be achieved for clinical cancer diagnostics., This article discusses the use of various Raman spectroscopy techniques and how they have been applied to different methods of detection for the diagnosis of cancer and in monitoring cancer treatment, with an outlook on how they can progress into a clinical environment.

Mg-based biodegradable materials, used for medical applications, have been extensively studied in the past decades. The in vitro cytocompatibility study showed that the proliferation and viability (as assessed by quantitative MTT-assay—3-(4,5-dimethyltiazol-2-yl)-2,5-diphenyl tetrazolium bromide) were not negatively affected with time by the addition of Mn as an alloying element. In this sense, it should be put forward that the studied alloys don’t have a cytotoxic effect according to the standard ISO 10993-5, i.e., the level of the cells’ viability (cultured with the studied experimental alloys) attained both after 1 day and 5 days was over 82% (i.e., 82, 43–89, 65%). Furthermore, the fibroblastic cells showed variable morphology (evidenced by fluorescence microscopy) related to the alloy sample’s proximity (i.e., related to the variation on the Ca, Mg, and Mn ionic concentration as a result of alloy degradation). It should be mentioned that the cells presented a polygonal morphology with large cytoplasmic processes in the vicinity of the alloy’s samples, and a bipolar morphology in the remote region of the wells. Moreover, the in vitro results seem to indicate that only 0.5% Mn is sufficient to improve the chemical stability, and thus the cytocompatibility; from this point of view, it could provide some flexibility in choosing the right alloy for a specific medical application, depending on the specific parameters of each alloy, such as its mechanical properties and corrosion resistance. In order to assess the in vivo compatibility of each concentration of alloy, the pieces were implanted in four rats, in two distinct body regions, i.e., the lumbar and thigh. The body’s reaction was followed over time, 60 days, both by general clinical examinations considering macroscopic changes, and by laboratory examinations, which revealed macroscopic and microscopic changes using X-rays, CT(Computed Tomography), histology exams and SEM (Scanning Electron Microscopy). In both anatomical regions, for each of the tested alloys, deformations were observed, i.e., a local reaction of different intensities, starting the day after surgery. The release of hydrogen gas that forms during Mg alloy degradation occurred immediately after implantation in all five of the groups examined, which did not affect the normal functionality of the tissues surrounding the implants. Imaging examinations (radiological and CT) revealed the presence of the alloy and the volume of hydrogen gas in the lumbar and femoral region in varying amounts. The biodegradable alloys in the Mg-Ca-Mn system have great potential to be used in orthopedic applications.

Ensilication is a novel method of protein thermal stabilisation using silica. It uses a modified sol–gel process which tailor fits a protective silica shell around the solvent accessible protein surface. This, electrostatically attached, shell has been found to protect the protein against thermal influences and retains its native structure and function after release. Here, we report the calorimetric analysis of an ensilicated model protein, hen egg-white lysozyme (HEWL) under several ensilication conditions. DSC, TGA-DTA-MS, CD, were used to determine unfolding temperatures of native, released and ensilicated lysozyme to verify the thermal resilience of the ensilicated material. Our findings indicate that ensilication protects against thermal fluctuations even at low concentrations of silica used for ensilication. Secondly, the thermal stabilisation is comparable to lyophilisation, and in some cases is even greater than lyophilisation. Additionally, we performed a mouse in vivo study using lysozyme to demonstrate the antigenic retention over long-term storage. The results suggest that protein is confined within the ensilicated material, and thus is unable to unfold and denature but is still functional after long-term storage.

Phage-based biosensors have shown significant promise in meeting the present needs of the food and agricultural industries due to a combination of sufficient portability, speed, ease of use, sensitivity, and low production cost. Although current phage-based methods do not meet the bacteria detection limit imposed by the EPA, FDA, and USDA, a better understanding of phage genetics can significantly increase their sensitivity as biosensors. In the current study, the signal sensitivity of a T4 phage-based detection system was improved via transcriptional upregulation of the reporter enzyme Nanoluc luciferase (Nluc). An efficient platform to evaluate the promoter activity of reporter T4 phages was developed. The ability to upregulate Nluc within T4 phages was evaluated using 15 native T4 promoters. Data indicates a six-fold increase in reporter enzyme signal from integration of the selected promoters. Collectively, this work demonstrates that fine tuning the expression of reporter enzymes such as Nluc through optimization of transcription can significantly reduce the limits of detection.

Quantitively and stably tracking ion dynamics in the living brain of animals is essential to understanding many physiological and pathological processes. Solid-state ion-selective electrodes (ISEs) are powerful tools for monitoring the dynamic change of ions at physiological concentration range; however, the unintentional accumulation of an aqueous layer at the ion selective membrane/ solid contact interface compromises the electrode potential stability, limiting its in vivo application. Here, using manganese dioxide (MnO2) and potassium ISE (K+-ISE) as model solid contact and ISEs, we demonstrate for the first time that graphdiyne oxide (GDYO) can enhance the potential stability of solid contact-based ISEs. Our results suggest that the intrinsic structural and hydrophobic features of GDYO, plays a key role in impeding and stabilizing the formation of water layer. With GDYO-MnO2 acting as the solid contact, the K+-ISE displays an excellent short-term potential stability and maintains great selectivity, achieving reliable K+ sensing at the animal level. The GDYO-based strategy is generalizable to different ISEs and does not require complicated processing steps, paving an exciting opportunity for in vivo ion recognition and sensing.

A microfluidic device as a pivotal research tool in chemistry and life science is now widely recognized. Indeed, microfluidic techniques have made significant advancements in fundamental research, such as the inherent heterogeneity of single-cells studies in cell populations, which would be helpful in understanding cellular molecular mechanisms and clinical diagnosis of major diseases. Single-cell analyses on microdevices have shown great potential for precise fluid control, cell manipulation, and signal output with rapid and high throughput. Moreover, miniaturized devices also have open functions such as integrating with traditional detection methods, for example, optical, electrochemical or mass spectrometry for single-cell analysis. In this review, we summarized recent advances of single-cell analysis based on various microfluidic approaches from different dimensions, such as in vitro, ex vivo, and in vivo analysis of single cells.

Sugar palm leaf stalk (petioles) ashes traditionally used as a daily cosmetic by Sundanese women to keep their skin smooth. Sugar palm leaf stalk ashes also used to treat acne, smallpox, and burns. The study was aimed to test the sunscreen activity of ashes extracts from sugar palm leaf stalk. The ashes were extracted using soxhlet and 96% ethanol as solvent. The sunscreen activity test was performed on the extract using UV-Vis spectrophotometer. Determination of Sun Protection Factor (SPF) value was done at 290-320 nm (UV-B wavelength) with 5 nm interval. The ashes extract of sugar palm leaf stalk showed the effectiveness of sunscreen activity at 10,000 ppm (SPF value = 9) as maximum protection. The SPF value was determined using Mansur mathematical equation. The determination of pigmentation transmittant was done at 322.5 – 372.5 nm, with 2.5 nm interval. Based on the result of pigmentation transmittance percentages, the sunscreen profile of sugar palm leaf stalk ashes extract (2000-10.000 ppm) showed the category of extra protection up to sunblock, according to classification from Balsam & Sagarin (1972). To be concluded, the ashes extract of sugar palm leaf stalk was a potential sunscreen. Further in vivo analysis should be done to confirm this potency.

Depending on their concentrations the surface-active substances, tensides (surfactants) can positively or negatively influence the drug absorption, which is widely used in the design of the dosage forms with controlled release. A problem is that the (in-vivo) rate of absorption cannot be directly measured and for that reason, it is frequently substituted by evaluation of the (in-vitro) dissolution. On other hand, a suitably designed pharmacokinetic model can directly predict virtually all pharmacokinetic quantities including both the rate of absorption and fraction of the dose reaching the blood circulation. The paper presents a new approach to the analysis of the rate of drug absorption and shows its superiority over traditional in-vivo approaches. Both the in-vivo analysis and model-based prediction of the tenside (monolaurin of sucrose) influence on the rate of absorption of the drug (sulfathiazole) after instantaneous per-oral administration to rats are discussed. It was found that 0.001% solution of tenside can increase the rate of absorption by cca 50% and a two-fold increase in absolute bioavailability can be reached. Attention is also devoted to the formal requirements laid on the model’s structure and its identifiability. The systematic design, substantiation and validation of a parsimonious predictive model that confirms in-vivo results are presented. The match between in-vivo observations and model-based predictions is demonstrated. The frequently overlooked metaphysics lying behind the compartmental modelling is briefly explained.

Light activation is an effective way to impart spatiotemporal control over oligonucleotide probes that are widely applied for gene expression regulation and target function investigation. Among the major oligonucleotide caging strategies, cyclization with a photocleavable linker is an elegant design, which affords both atom efficiency and stability in many biological environments. Here, we introduce an improved protocol for circular oligonucleotide synthesis requiring only one round of HPLC purification. With a series of poly-U oligonucleotide strands of different sizes and backbone modifications, the pre-photolysis caging stability and post-photolysis target binding affinity were studied through a denaturing gel assay and melting temperature measurements. A 14U 2’-OMe RNA probe was selected, with strong potential application in transcriptome in vivo analysis (TIVA) for mRNA isolation.

Background: Nature has always been considered as the primary source of pharmaceutical ingredients. A variety of hemicelluloses, as well as their modified forms, have been under investigation. Herein, a study was designed to explore the biocompatibility of hemicellulose and its modified form (thiolated hemicellulose) as well as its potential as a pharmaceutical excipient.Method: For thiol modification thiourea was used as the thiol donor, HCl as the catalytic reagent, and methanol was used for washing purposes. Modified polymers were characterized for physicochemical characteristics, including surface morphology, the amorphous or crystalline nature of the particles, modification of polymer by FTIR, and biocompatibilities. For acute oral toxicity study, a single dose of 2 g/kg was administered to albino rats of 200 g average weight (n = 3). Polymers were evaluated as pharmaceutical excipients by preparing compressed tablets of antiplatelet drug (Ticagrelor), followed by various quality control tests, such as swelling index, thickness and diameter, disintegration, and in-vitro drug release.Results: From the results, it was observed that thiol modification has been successfully accomplished as characteristic peaks belonging to –SH group appeared at 2667.7691 cm−1 in FTIR scan. The modified polymer was found safe in the use concentration range, confirming their safe use for in vivo analysis. No significant effect has been observed in the behavior, biological fluid (blood), or on vital organs. Thiolated hemicellulose was found to be an excellent drug retarding polymer as 8 h of dissolution studies showed that 67.08% of the drug has been released.Conclusion: Conclusively, incorporation of thiol moiety made the polymer more mucoadhesive with, and a worthy carrier of, the drug with good biocompatibilities.

The current study was designed to convert the poloxamer (PLX) into thiolated poloxamer (TPLX), followed by its physicochemical, biocompatibilities studies, and applications as a pharmaceutical excipient in the development of tacrolimus (TCM)-containing compressed tablets. Thiolation was accomplished by using thiourea as a thiol donor and hydrochloric acid (HCl) as a catalyst in the reaction. Both PLX and TPLX were evaluated for surface morphology based on SEM, the crystalline or amorphous nature of the particles, thiol contents, micromeritics, FTIR, and biocompatibility studies in albino rats. Furthermore, the polymers were used in the development of compressed tablets. Later, they were also characterized for thickness, diameter, hardness, weight variation, swelling index, disintegration time, mucoadhesion, and in vitro drug release. The outcomes of the study showed that the thiolation process was accomplished successfully, which was confirmed by FTIR, where a characteristic peak was noticed at 2695.9968 cm−1 in the FTIR scan of TPLX. Furthermore, the considerable concentration of the thiol constituents (20.625 µg/g of the polymer), which was present on the polymeric backbone, also strengthened the claim of successful thiolation. A mucoadhesion test illustrated the comparatively better mucoadhesion strength of TPLX compared to PLX. The in vitro drug release study exhibited that the TPLX-based formulation showed a more rapid (p &lt, 0.05) release of the drug in 1 h compared to the PLX-based formulation. The in vivo toxicity studies confirmed that both PLX and TPLX were safe when they were administered to the albino rats. Conclusively, the thiolation of PLX made not only the polymer more mucoadhesive but also capable of improving the dissolution profile of TCM.

The vegetal world constitutes the main factory of chemical products, in particular secondary metabolites like phenols, phenolic acids, terpenoids, and alkaloids. Many of these compounds are small molecules with antibacterial activity, although very few are actually in the market as antibiotics for clinical practice or as food preservers. The path from the detection of antibacterial activity in a plant extract to the practical application of the active(s) compound(s) is long, and goes through their identification, purification, in vitro and in vivo analysis of their biological and pharmacological properties, and validation in clinical trials. This review presents an update of the main contributions published on the subject, focusing on the compounds that showed activity against multidrug-resistant relevant bacterial human pathogens, paying attention to their mechanisms of action and synergism with classical antibiotics.

International audience; The study of skin mechanical impedance is a clue to analyse its health. We use a Langevin Transducer at 60kHz to excite the skin. In return we observe the acoustic forces generated by the interaction. To achieve that, the system is identified and presented in the rotating reference frame, and we describe the design of two PI controllers that accurately control the vibration velocity. However, in vivo analysis can be tricky when it comes to safety and duration of tests. Thus, phantom tissues are used as substitute to simulate skin behaviour. Due to its mechanical properties similar to skin, and its simple manipulation, Dragon Skin ®, a silicone rubber type of phantom, was chosen. The paper presents the characterisation of the mechanical impedance of three silicon phantoms with different properties based on force observation. The results show that the phantoms can be indeed identified based on this method.

The photoreceptor-specific nuclear receptor Nr2e3 is not expressed in Nr2e3

The study sought to formulate and evaluate suppositories using a locally produced brand of alum (Aw) obtained from bauxite waste generated at Awaso bauxite mine in the Western-North region of Ghana, for use in the treatment of hemorrhoids. The suppositories were formulated using shea butter modified, respectively, with amounts of beeswax and theobroma oil. In another development, theobroma oil was modified with different concentrations of beeswax. Drug-base interactions were investigated using attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy. The suppositories were prepared using the hot melt and trituration methods. Quality control checks were carried out on the formulations. The evaluated parameters included physical characteristics (texture, presence or absence of entrapped air, and contraction holes), weight uniformity, disintegration time, drug content, and in vitro release profile of the alum from the formulated suppositories. An in vivo analysis was carried out on the most suitable formulation to ascertain its efficacy on inflamed tissues using croton oil-induced rectal inflammation in a rat model. A critical examination of the ATR-FTIR spectra revealed no drug-base interactions. The suppository formulations passed all Pharmacopoeia stated tests. The in vivo study revealed the use of suppositories ameliorated the croton oil-induced hemorrhoid in the rectoanal region of the rats.

In most organisms, methylglyoxal (MG), a toxic metabolite by-product that causes diabetes in humans, is predominantly detoxified by the glyoxalase enzymes. This process begins with the so-called “spontaneous” conjugation of MG with the cytoprotectant metabolite glutathione (GSH). In this study, we unravel a logical, but as yet unsuspected, link between MG detoxification and a (prokaryotic) representative of the ubiquitous glutathione transferase (GST) enzymes. We show that a GST of a model cyanobacterium plays a prominent role in the detoxification of MG in catalyzing its conjugation with GSH. This finding is important because this reaction, always regarded as nonenzymatic, could exist in plants and/or human and thus have an impact on agriculture and/or human health., Methylglyoxal (MG) is a detrimental metabolic by-product that threatens most organisms (in humans MG causes diabetes). MG is predominantly detoxified by the glyoxalase pathway. This process begins with the conjugation of MG with glutathione (GSH), yielding a hemithioacetal product that is subsequently transformed by the glyoxalase enzymes into d-lactate and GSH. MG has been overlooked in photosynthetic organisms, although they inevitably produce it not only by the catabolism of sugars, lipids, and amino acids, as do heterotrophic organisms, but also by their active photoautotrophic metabolism. This is especially true for cyanobacteria that are regarded as having developed photosynthesis and GSH-dependent enzymes to detoxify the reactive oxygen species produced by their photosynthesis (CO2 assimilation) and respiration (glucose catabolism), which they perform in the same cell compartment. In this study, we used a combination of in vivo and in vitro approaches to characterize a logical, but as yet never described, link between MG detoxification and a (prokaryotic) representative of the evolutionarily conserved glutathione transferase (GST) detoxification enzymes. We show that the Sll0067 GST of the model cyanobacterium Synechocystis sp. strain PCC 6803 plays a prominent role in MG tolerance and detoxification, unlike the other five GSTs of this organism. Sll0067 catalyzes the conjugation of MG with GSH to initiate its elimination driven by glyoxalases. These results are novel because the conjugation of MG with GSH is always described as nonenzymatic. They will certainly stimulate the analysis of Sll0067 orthologs from other organisms with possible impacts on human health (development of biomarkers or drugs) and/or agriculture.

Background: Rosa damascena Mill, commonly known as Damascus rose and garland rose, belongs to Rosaceae family, which has around 200 different types of rose species. R. damascena Mill is famous for its fragrance and beauty, and hence it is highly cultivated all over the world. In addition, this plant is also used for its medicinal properties. Objectives: The objectives of this study are to prepare mouth wash and gel from the oil of R. damacena Mill, to evaluate the effectiveness of the medicinal properties of prepared mouth wash and gel, to evaluate its anti-inflammatory and analgesic properties, and to evaluate the adverse effects. Materials and Methods: At the JSS College of Pharmacy, Ooty, the oil of R. damacena Mill was converted into mouth wash and gel. To evaluate the effectiveness of the prepared rose gel and mouthwash, 24 Wister and 24 Albino rats were subjected to acetic acid induced writhing, hot plate method, and Carragennin induced paw edema test. Results: Acetic acid induced writhing in mice and hot plate method were carried out. Both methods proved presence of analgesic activity, and Carragennin induced paw edema test showed the anti-inflammatory property of R. damascena mill. Conclusion: These two in vivo studies proved that the presence of analgesic and anti-inflammatory properties in prepared R. Damascena mill mouth wash and gel.

WOS: 000462070300008, PubMed: 30540620, Objective Formation mechanisms and treatment of the urinary stones are different, depending on their chemical structure. Therefore, determining the stone type plays a key role in planning treatment and preventive measures. Computed tomography (CT), with the use of dual-energy technology in recent years, has made it possible to do in vivo analysis of urinary stones. In this study, we aimed to evaluate the diagnostic efficacy of dual-energy CT (DECT) and compare its results with in vitro analysis, which is accepted as a gold standard for analysis of urinary stones. Materials and Methods The DECT examinations were performed on 373 patients using 128-slice dual-source CT scanner. Analysis of attenuation ratios in the high and low kilovoltage peak values of the stone was performed at workstation, and stones were classified as hydroxyapatite, calcium oxalate, cystine, and uric acid. On follow-up, the stone was obtained in 35 patients as a result of surgery or passed spontaneously. The DECT analysis and in vitro analysis results were compared and statistically evaluated. Results In all patients, 136 hydroxyapatite, 160 calcium oxalate, 57 uric acid, and 20 cystine stones were detected with DECT. In vitro analyses of the stones were performed in 35 patients, and 8 hydroxyapatite, 18 calcium oxalate, 6 uric acid, and 3 cystine stones were revealed. When DECT analysis results were compared with in vitro analysis results, stone types were detected correctly in 32 (91.4%) patients and incorrectly in 3 (8.6%) patients. Especially all uric acid and cystine stones were correctly detected with DECT. Conclusions With advanced postprocess analysis methods, DECT is able to analyze urinary stones. The DECT is found superior especially in detecting uric acid and cystine stones. Its success in detecting hydroxyapatite and calcium oxalate stones is also high. When in vivo analyses of the stones are performed with DECT, it will be possible to make a contribution to the personalization and optimization of the treatment.

Increased global incidence of myopia necessitates establishment of therapeutic approaches against its progression. To explore agents which may control myopia, we screened 207 types of natural compounds and chemical reagents based on an activity of a myopia suppressive factor, early growth response protein 1 (Egr-1) in vitro. Among the candidates, crocetin showed the highest and dose-dependent activation of Egr-1. For in vivo analysis, experimental myopia was induced in 3-week-old C57BL/6 J mice with −30 diopter (D) lenses for 3 weeks. Animals were fed with normal or mixed chow containing 0.003% (n = 19) and 0.03% (n = 7) of crocetin during myopia induction. Refraction and axial length were measured at 3-week-old and the 6-week-old with an infrared photorefractor and a SD-OCT system. Compared to controls (n = 14), crocetin administration showed a significant smaller change of refractive errors (−13.62 ± 8.14 vs +0.82 ± 5.81 D for 0.003%, p

In this study, we developed contactless electrospray ionization mass spectrometry (ESI-MS) for in vivo analysis of living organisms in different applications. The in vivo sampling and direct analysis processess of living organisms were integrated into an operation that only requires the organism close to MS inlet that was applied to a high voltage. Living plants and animals were directly induced to generate spray ionization. Direct detection and in vivo monitoring of metabolites and chemical residues in various living organisms were successfully demonstrated. Analysis of a single sample could be completed within 30 s. Overall, contactless ESI-MS provides an attractive in vivo method to straightforward investigation of living organisms.

Noodles are widely consumed in China, which can be cooked in different ways. The effects of different cooking methods (boiling, steaming, microwave heating, stir-frying and frying) on the resistance starch (RS) content and digestive properties (digestion rate, digestibility and estimated glycemic index (eGI) value) of noodles were investigated. The RS content was greatly affected by the cooking time, and it was varied when the noodles were optimally cooked using different cooking methods. The RS contents of the microwaved and stir-fried noodles were relatively high (0.59%&ndash, 0.99%), but it was lower (0.43%&ndash, 0.44%) in the boiled and steamed noodles. Microwaved noodles showed the slowest digestion rate and the lowest eGI. Due to the limited water within fried noodles, none RS was found in the fried noodles, whereas stir-fried noodles showed RS5 formation from the XRD and DSC results. Compared with boiled and steamed noodles, the microwaved noodles showed a more compact morphology without porous holes on the surface, whereas fried noodles showed irregular morphology. The results indicated that the digestive properties of noodles made with the same ingredients can be greatly altered by using different cooking methods, and the digestive properties of different cooked noodles are worthy of confirmation using in vivo analysis.

Purpose The aim of the present in vivo analysis was to evaluate the osseointegration process of titanium implants with three different surfaces (machined, sandblasted and acid-etched, and laser-treated) after 15 and 30 days of healing period. Materials and methods Thirty-six implants with different surfaces were placed in the iliac crest of four Bergamasca sheep. The implant surfaces tested were sandblasted and acid-etched (group A), laser-treated (group B), and fully machined (group C). Two animals were sacrificed after 15 days while the other two after 30 days. Histological and histomorphometric analyses were performed. Results After 30 days, the bone tissue layer onto implant groups A and B appeared almost continuous with small marrow spaces interruption, while on the machined surface (group C), larger spaces with marrow tissue alternated with the bony trabeculae onto the titanium surface. Implants in groups A and B showed significantly higher implant contact percentage (BIC%) value than group C (P < 0.05). Moreover, it was observed a BIC% increase in both groups A and B between 15 and 30 days while in the machined group (group C), the BIC% decreased. Conclusion Results from the present in vivo analysis revealed that both sandblasted/acid-etched and laser-treated titanium implants, compared to the machined ones, have higher values of osseointegration in less healing time.

Over the past two decades, there has been increasing focus on ascorbic acid (AA) due to its anti-oxidant and neuroprotective properties as well as its neuromodulating capability. Conventional analytical methods for selective in vivo monitoring of AA mainly involve complex procedures, which lower the temporal resolution and throughput of data gathering. Moreover, analytical methods for selective real-time monitoring of AA exocytosis at a single-cell level is still lacking. The lack of effective methods for AA detection in the central nervous system (CNS) has rendered difficulties in better understanding the roles of AA in brain function. AA is, in itself, electrochemically active, and thereby rationally tailoring the structure of an electrode/solution interface would offer an effective approach to selective electrochemical measurements in the CNS. Guided by this, electrochemical methods have been recently established for AA detection by combining selective electrochemical oxidation of AA at functionalized electrodes with microelectrode techniques and with in vivo microdialysis. This review mainly focuses on recent updates on in vivo detection of AA by modulating the electron transfer of AA to achieve the selectivity for its detection in the CNS, an environment with high chemical complexity. Additionally, the practical implications of the methods in selective and sensitive monitoring the dynamics of AA in different brain functions are also reviewed.

Purpose To prospectively assess the ability of Dual-energy CT (DECT) for determining the composition of urinary tract calculi in vivo. Subjects and methods 60 patients (34 men and 26 women; age range 22–68 years; mean age 42 ± 10.3 years) with urolithiasis were examined with DECT using a protocol that included 100/Sn140 kV. The calculi were categorized as uric acid, cysteine, and calcium oxalate or phosphate in accordance with their attenuation values at 100/Sn140 kV. Chemical analysis of the calculi was used as reference standard. Results 84 calculi were identified in 60 patients (58 calculi were found in renal calyces, 14 in ureters, 10 in the renal pelvis, and two in the bladder). The DECT correctly revealed calculus composition in 76/84 (90.5%) of the patients. The DECT is highly reliable in discriminating uric acid from non-uric acid calculi in 82/84 (97.6%) of the patients. The sensitivity and specificity in distinguishing uric acid from non-uric acid calculi were 100% and 96.9% respectively. Calcified calculi were difficult to be discriminated. Mixed calculi were excluded from the study. Conclusion DECT allow the correct description of urinary calculus composition in vivo.

Despite their functional and structural diversity, membrane proteins of all living cells are inserted and assembled via a few evolutionary conserved pathways. Here, we describe how studies in model membranes provide insights into the functionality of the Sec translocon, a primary machinery for the insertion of a-helical membrane proteins, but also for translocation of unfolded polypeptide chains across the membrane. To bring together in vitro and in vivo analysis, further developments of new membrane mimetics are suggested.

WOS: 000432561200003 The purpose of this study is the in vivo analysis of the changes in iNOS expression in the ovaries of the capsaicin-treated rats in puberty and adult periods. Rats were divided into with two age groups i.e. puberty (42 days old) and adult (70 days old). Each age group was divided into three sub-groups as treated experimental or vehicle, and untreated control. Starting from 21 days up to 42 and 70 days, implementations were as follows: experimental group received a daily subcutaneous injection of capsaicin (1mg/kg body weight); vehicle group received the vehicle solution (10% Tween 80, 10% ethanol, 80% distilled water) in same volume as experimental group, and untreated control group was not subjected to any injections. In ovaries of all groups, iNOS expression was shown in granulosa cells, interstitial cells, theca follicle cells and corpus luteum luteal cells. An immune reaction was observed in pubertal and adult rat ovaries and iNOS was not inactivated with age. The reaction density was reduced in the experimental groups and showed long-term capsaicin implementation in low dosage negatively affects iNOS stimulation in ovaries.

Nitrogen and phosphorus are macronutrients indispensable for plant growth. The acquisition and reallocation of both elements require a multitude of dedicated transporters that specifically recognize inorganic and organic forms of nitrogen and phosphorous. Although many transporters have been discovered through elegant screening processes and sequence homology, many remain uncharacterized for their functions in planta. Genetically encoded sensors for nitrogen and phosphorous molecules offer a unique opportunity for studying transport mechanisms that were previously inaccessible. In the past few years, sensors for some of the key nitrogen molecules became available, and many improvements have been made for existing sensors for phosphorus molecules. Methodologies for detailed in vivo analysis also improved. We summarize the recent improvements in genetically encoded sensors for nitrogen and phosphorus molecules, and the discoveries made by using such sensors.

Summary With a long history in research, bioelectrochemistry has emerged as a hot spot in the development of implantable biosensors with desired selectivity, sensitivity and spatiotemporal resolution for accurate analysis of electroinactive species in vivo . To catalyze electrode reactions (oxidation or reduction) of target analytes, enzymes are employed as the efficient bioelectrocatalysts immobilized on the electrodes. In bioelectrochemistry, the biggest challenge is the interfacial electron transfer between the redox active centers of enzymes and electrode surfaces. Besides, enzymatic electrodes are prone to inhibition and inactivation by environmental factors (co-substrates, inhibitors, pH, etc.). Rational design of the electrode surface/interface is necessitated to fulfill the requirements of in vivo applications. This mini review covers the recent and representative progress on interface engineering to enable bioelectrochemistry for in vivo analysis with an emphasis on heterogeneous electron transfer, oxygen dependence and microenvironment tolerance of bioelectrodes and self-powered electrochemical biosensors as well.

Tissue membranes are boundaries that isolate organs or cavities in the body. These semi-permeable membranes are responsible for passive protection that acts through the regulation of nutrient absorption, secretion and filtration of small molecules. These functions could be altered as a consequence of inflammation or trauma, which in turn could lead to changes in permeability, allowing the entrance of toxins, antigens, proteins or facilitating the spread of tumors. Membrane permeability therefore plays an important role in numerous diseases. However, current experimental techniques that are available to quantify membrane permeability in small animals have limited precision and temporal specificity. Improvements in such measurements would lead to a deeper understanding of disease pathogenesis and this may accelerate the development of specific therapies. The study reported here concerns the efficacy of a novel, non-invasive imaging analysis-based measurement method that significantly improves the quantification of tissue membrane permeability in small animals, while at the same time mitigating the adverse effects experienced by the animals under study.

Three-dimensional (3D) porous scaffolds for supporting cell adhesion and growth play a vital role in tissue engineering applications. In the present study, three different collagen-based 3D sponges were functionalized by apatite coating. The sponges were coated with apatite on their outer and inner surfaces while retaining their interconnecting pores. To achieve this, we employed a vacuum degassing system in our plasma- and precursor-assisted biomimetic process using a supersaturated calcium phosphate solution. The resulting apatite-coated sponges (mineralized sponges) showed better cell adhesion properties in vitro for osteoblast-like MC3T3-E1 cells compared to that of uncoated sponges. The three mineralized sponges were implanted in the subcutaneous tissue of rats. Upon histological evaluation after 10 days, the mineralized sponges showed cell in-growth rates that were approximately 4-fold greater than those of the untreated sponges without any notable inflammatory reactions. As these sponges are composed of clinically approved collagen-based frameworks and possess a 3D porous structure with a mineralized surface appropriate for cell adhesion and internalization, further in vitro and in vivo studies should be conducted regarding tissue engineering applications.

Measuring the kinetics of the violaxanthin cycle imposes an experimental challenge. Traditionally, carotenoid analysis was carried out laboriously with high-performance liquid chromatography. In this work, we present the first in vivo approach to directly measure the kinetics of the violaxanthin cycle, using resonance Raman spectroscopy in combination with baseline correction and principal component analysis. Applying the new approach allows measuring thousands of data points as opposed to the few possible with chemical analysis over the course of a violaxanthin cycle kinetics experiment. In vivo analysis of the violaxanthin cycle is necessary to fully understand adaptation kinetics to varying light conditions, the knowledge of which is especially important for assessing the stress tolerance of plants in the wake of the increasing climate change. Three experiments on the green alga Dunaliella salina were performed, featuring both the light-to-dark and dark-to-light transitions response of the algae. Copyright © 2017 John Wiley & Sons, Ltd.

With the ever-increasing human lifespan, age-related affections have become a public health issue. The health sector is looking for new bioactive compounds to respond to this demand. The unexplored microbial biodiversity and its metabolites represent a major source of innovative bioactive molecules with health potential. Fermented foods, such as raw-milk cheese, have already been investigated for their rich microbial environment, especially for their organoleptic qualities. But studies remain limited regarding their effects on health and few metabolites of microbial origin have been identified. An efficient methodology was developed in this study to investigate the biological effect of raw-milk cheese, combining a chemical fractionation, to isolate the most metabolites from the cheese matrix, and anin vivobiological test usingCaenorhabditis elegans.C.eleganswas brought into contact with cheese extracts, obtained by means of chemical fractionation, and with freeze-dried whole cheese by supplementing the nematode growth medium. A longevity assay was performed to evaluate the effects of the extracts on the worms. Our results demonstrate the feasibility of the method developed to bring the worms into contact of the cheese extracts. The evaluation of the effects of the extracts on the longevity was possible. Some extracts showed a beneficial effect as extract W70 for example, obtained with water, which increases the mean lifespan by 16% and extends the longevity by 73% (p< 0.0001).

Ambient ionization mass spectrometry (AIMS) has grown into a group of emerging analytical techniques that allow rapid, real-time, high-throughput, in situ, and in vivo analysis in many scientific fields including biomedicine, pharmaceuticals, and forensic sciences. While dozens of AIMS techniques have been introduced over the past two decades, their broad commercial and industrial use is still restricted by multiple challenges. In this Perspective, we discuss the most relevant technical challenges facing AIMS, i.e., reproducibility, quantitative ability, molecular coverage, sensitivity, and data complexity, and scientists' recent attempts to overcome these hurdles. Furthermore, we present future directions of AIMS from our perspective, including the necessity that efforts should be made to unravel blind biomolecules in routine analysis, the construction of a data depository for AIMS users, the full automation of pipelines for prospect integration in a robotic laboratory, the movement toward on-site tests, and the expansion of outreach to motivate government officials in policymaking. We anticipate that, with progress in these critical but immature areas, AIMS technology will keep evolving to become a more robust and user-friendly set of technologies and, consequently, be translated into everyday life practice.

Made available in DSpace on 2020-12-12T01:16:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-02-01 X-ray fluorescence spectroscopy (XRF) is an analytical tool used to determine the elemental composition in a myriad of sample matrices. Due to the XRF non-destructive feature, this technique may allow time-resolved plant tissue analyses under in vivo conditions, and additionally, the combination with other non-destructive techniques. In this study, we employed handheld and benchtop XRF to evaluate the elemental distribution changes in living plant tissues exposed to X-rays, as well as real-time uptake kinetics of Zn(aq) and Mn(aq) in soybean (Glycine max (L.) Merrill) stem and leaves, for 48 hours, combined with transpiration rate assessment on leaves by an infrared gas analyzer (IRGA). We found higher Zn content than Mn in stems. The latter micronutrient, in turn, presented higher concentration in leaf veins. Besides, both micronutrients were more concentrated in the first trifolium (i.e., youngest leaf) of soybean plants. Moreover, the transpiration rate was more influenced by circadian cycles than Zn and Mn uptake. Thus, XRF represents a convenient tool for in vivo nutritional studies in plants, and it can be coupled successfully to other analytical techniques. Center for Nuclear Energy in Agriculture (CENA) University of Saõ Paulo (USP), Avenida Centenário, 303 Saõ Paulo State University-UNESP, Rua Domingos da Costa Lopes 780 Saõ Paulo State University-UNESP, Rua Domingos da Costa Lopes 780

Present study was conducted to optimize the processing parameters for galactooligosaccharides (GOS) synthesis from whey powder followed by exploring its prebiotic efficiency.All factors (initial lactose concentration, pH, reaction time, temperature and enzyme to substrate ratio; E/S) were analyzed by single factor analysis and optimization for GOS yield was done following the orthogonal experimental design. For in vivo analysis, 60 mice were equally divided into four groups (normal control, NC; low, medium, and high dose of GOS, LG, MG and HG) and fed varying levels (0, 0.25, 0.5 and 1.0 g/kg bw per day) of GOS, for 30 days and sampling was done at the end of experiment regarding gut health, immunity, cecal microbiota and metabolites.Optimum yield of GOS (25.1%) was obtained at reaction time 25 min, temperature 50 °C, pH 4.5 and the enzyme to substrate ratio (E/S) of 20 U/g. In vivo experiment, shallower crypt and greater villus to crypt ratio (V/C) were found in the duodenum of LG treatment compared to NC mice (P .05). The GOS promotes thymus development and improve immunity. Intervention with GOS increased the population of bifidobacterium and lactobacillus in MG and bifidobacterium in LG mice (P .05), and was accompanied by decreased proliferation of desulfovibrio. Correlation analysis also supported the above findings.This study optimized the processing parameters for GOS preparation and provided data encouraging to suggest that GOS can be a potential option to improve the gut health and immunity.

Neurotransmitters (NTs) constitute an important group of messenger molecules and their imbalance lead to various neurological disorders, making their analytical determination of great importance in both laboratory and clinical practice. Here we review the most recent progress in sample pretreatment and in vivo analysis for various NTs and metabolites focusing on two approaches: biosensors and SPME, which require small amounts of biological samples and have wide application. Biosensors, as integrated analytical tools, provide the chance for direct monitoring of NTs and their dynamics directly in a tissue. In turn, non-exhaustive SPME method enables a high-throughput and effective extraction of endogenous compounds like NTs, with minimal invasiveness, which is of particular importance for in vivo analysis. Hence, these techniques are very promising and warrant application and further development.

Nowadays, mastitis is one of the biggest problems in breeding dairy cattle. Treatment of this disease with conventional antibiotics is ineffective because many pathogens are resistant. Researchers have therefore been forced to look for new solutions, and metal nanoparticles (NPs) have been found to be the most appropriate agents. This study uses commercially available silver (AgNPs) and copper (CuNPs) nanoparticles and synthetized silver&ndash, copper nanoparticles (AgCuNPs) to evaluate the effect of these NPs on human and bovine mammary cells. The effect of AgNPs, CuNPs, and AgCuNPs on pathogen species commonly involved in udder inflammation (e.g., Staphylococcus aureus and Escherichia coli) was also established. The results show that commercially available NPs were of good quality and did not have a toxic effect on mammary gland tissue. AgNPs, CuNPs, and AgCuNPs also influenced or decreased the viability of pathogens. Therefore, the presented data suggest that metal NPs could be used in mastitis prevention and treatment in the future. However, the presented preliminary results require further in vivo analysis.