Testing In Vivo

Long exposure to small concentrations of xenoestrogens (XEs) seems to be responsible for the increase of certain hormone-dependent diseases. The results of the research concerning the study of potentially synergic interactions between XEs are few and contradictory and need further studies. The oxidative stress is involved in the generation of several pathologies, such as cancer. We studied the oxidative stress induction in rats after exposure to bisphenol A (BPA) and/or methylparaben, two xenobiotics identified as XEs. Malondialdehyde (MDA), a lipid peroxidation end-product, and 2,3-dihydroxybenzoic acid (2,3-DHBA), a marker of in vivo hydroxyl radical production, were used to evaluate the oxidative stress generation. The levels of MDA in rat plasma were determined by HPLC-UV (detection at 307 nm) after derivatization with dinitrophenylhydrazine. 2,3-DHBA was analyzed by LC-MS/MS in multiple reaction monitoring (MRM) mode using an ion trap mass spectrometer with electrospray negative ionization. The ion transition monitored was m/z 153 → m/z (109). The results indicated that both studied xenobiotics have the potential of oxidative stress induction in rats but simultaneous exposure does not increase the intensity of effects. producerii de radicali hidroxil in vivo. Nivelele de MDA în plasma de şobolan s-au determinat prin HPLC-UV (detecţie la 307 nm) după derivatizare cu dinitrofenilhidrazină, iar cele de 2,3-DHBA prin LC-MS/MS în modul MRM (multiple reaction monitoring) folosind un spectrometru de masă cu trapă ionică şi ionizare negativă prin electrospray. S-a monitorizat tranziţia m/z 153 → m/z (109). Rezultatele au indicat faptul că ambele xenobiotice studiate au potenţial de a induce stres oxidativ la şobolani, dar în cazul expunerii simultane nu s-a observat o creştere a intensităţii efectelor.

In vivo multiphoton tomography with a wavelength-tunable femtosecond laser has been performed to investigate the autofluorescence intensity of major endogenous fluorophores of human skin in dependence on the excitation wavelength. In high-resolution multiphoton images of different skin layers, clear trends were found for fluorophores like keratin, NAD(P)H, melanin as well as for the elastin and collagen networks. The analysis of the measurements is supplemented by additional measurements of fluorescence lifetime imaging and signal-decay curves by time-correlated single-photon counting.

The aim of this research is to synthesize submicron-sized calcium carbonate vaterite particles of elliptical shape with different reaction durations. We also aim to assess their antioxidant and anti-inflammatory properties, which may be beneficial for treating diseases such as asthma and rheumatoid arthritis. Methods: Calcium carbonate vaterite particles were prepared using the co-precipitation method with varying reaction times, characterized by Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), Fourier Transformation Infrared Spectroscopy (FTIR), and Poly-Dispersity Index, with antioxidant activity assessed by the 2,2-Diphenyl-1-Picrylhydrazyl (DPPH) method and anti-inflammatory activity by the protein denaturation method. Results: SEM and TEM analysis revealed that the synthesized vaterite particles had an elliptical shape with nano-crystalline particles of around 50 nm size. FTIR verified the production of vaterite particles. Research on antioxidants and anti-inflammatory agents revealed that the crystalline particles exhibited DPPH scavenging action, with an IC50 of 12.6 µg/ml, and a noteworthy reduction in protein denaturation in the albumin protein denaturation test, with an IC50 of 222.49 µg/ml, in comparison to the reference value. Conclusion: The results highlight the potential of elliptical vaterite submicron micro-particles as versatile platforms with anti-inflammatory and antioxidant properties, paving the way for future advancements in drug delivery systems, food additives, and natural supplements by efficiently encapsulating drugs and proteins.

Luminescent nano‐thermometry is a fast‐developing technique with great potential for in vivo sensing, diagnosis, and therapy. Unfortunately, it presents serious limitations. The luminescence generated by nanothermometers, from which thermal readout is obtained, is strongly distorted by the attenuation induced by tissues. Such distortions lead to low signal levels and entangle absolute and reliable thermal monitoring of internal organs. Overcoming both limitations requires the use of high‐brightness luminescent nanothermometers and adopting more complex approaches for temperature estimation. In this work, it is demonstrated how superbright Ag2S nanothermometers can provide in vivo, reliable, and absolute thermal reading of the liver during laser‐induced hyperthermia. For that, a new procedure is designed in which thermal readout is obtained from the combination of in vivo transient thermometry measurements and in silico simulations. The synergy between in vivo and in silico measureme...

The chemical structure of human body odorants and the enzymatic pathways for their release from amino acid conjugates by skin bacteria have been elucidated in recent years. These findings open up theoretical avenues for an added layer of malodour prevention by blocking the cleavage of malodour precursors. Previously, we had validated in vitro an alternative substrate that releases a fragrance molecule instead of a malodour acid upon the action of a corynebacterial N a-acyl-glutamine-aminoacylase. In order to assess whether this concept delivers a perceivable reduction of malodour in vivo, a number of studies were performed: (1) a controlled clinical study with expert assessment, (2) a blinded study with consumer self-assessment of a placebo versus an active formulation in left-right comparison, and (3) a longitudinal consumer study comparing two doses of the active versus a placebo formulation. A statistically significant benefit for malodour reduction was observed in all three studies, this benefit consistently seen on panellists with a higher self-or expert-perceived malodour, whereas there was no difference in panellists with a lower level of malodour formation. In addition, malodour formation was higher in panellists with noticeable cleavage of the alternative substrate, indicating a correlation between enzyme activity and malodour levels. These results indicate that the targeted enzyme is a relevant source of malodour on panellists suffering from strong body odour, and that malodour reduction on these panellists is possible by specifically targeting this enzyme. Whereas the absolute reduction of malodour with the tested substrates was relatively small, the studies presented here clearly validate N a-acyl-glutamine-aminoacylase as a target for deodorant actives, and they demonstrate that a significant benefit can be achieved especially in individuals developing stronger axillary odour.

Selective laser melting (SLM) can produce complex hierarchical architectures paving the way for highly customisable biodegradable load-bearing bone scaffolds. For the first time, an indepth analysis on the performance of SLM-manufactured iron-manganese bone scaffolds suitable for load-bearing applications is presented. Microstructural, mechanical, corrosion and biological characterisations were performed on SLM-manufactured iron-manganese scaffolds. The microstructure of the scaffold consisted primarily of γ-austenite, leading to high ductility. The mechanical properties of the scaffold were sufficient for load-bearing applications even † CX and DC contributed equally to this work.

This study aimed to determine the impact of sesame meal, selenium (Se), and vitamin E (VitE) on goats’ oxidative status. Thirty mid-lactation crossbred goats were divided into five homogeneous groups, and were fed 1 kg of alfalfa hay and 1.2 kg of concentrates daily. The control group (C) received a basal diet. In the concentrates of the treated groups, 10% of the soybean meal was replaced by sesame meal and no extra VitE or Se (SM), or an extra 60 mg of VitE (SME), or 0.1 mg organic Se (SMSe), or their combination (60 mg VitE and 0,1 mg organic Se/kg of concentrate (SMESe). In the plasma of the goats, the dietary treatments did not affect glutathione reductase, glutathione peroxidase, glutathione transferase, catalase, superoxide dismutase activities, malondialdehyde (MDA) content, or the total antioxidant capacity. A reduction and a trend for lower protein carbonyls content was found in goats fed SM (p = 0.03) and SME (p = 0.06) compared to SMESe. In the milk, the lactoperoxidase ...

In this present study, the antioxidant properties of kefir produced from goat milk with kefir grains were investigated. The antioxidant capacity of kefir was evaluated by assessing the DPPH (2,2-Diphenyl-1-picrylhydrazyl), the ABTS-based method [2,2'-azino-bis-(3-ethylbenzthiazoline-6sulphonic acid)] radical-scavenging activity and ferric reducing antioxidant power (FRAP) at different stages of fermentation and storage period. Generally, the antioxidant capacity of goat milk-kefir samples was mainly dependent on the fermentation and storage period and good stability in DPPH, ABTS and FRAPS assays. During fermentation and storage, the total phenolic content in samples demonstrated significantly decreased.

Facial-expression-based analysis has been widely applied as a pain coding system in horses. Herein, we aimed to identify pain in horses undergoing subcutaneously polylactide-based polymer implantation. The sham group was submitted only to surgical incision. The horses were filmed before and 24 and 48 h after implantation. Five statistical methods for evaluating their facial expressions (FEs) were tested. Primarily, three levels of scores (0, 1, and 2) were applied to the seven FEs (ear movements, eyebrow tension, orbicularis tension, dilated nostrils, eye opening, muzzle tension, and masticatory muscles tension). Subsequently, the scores of the seven FEs were added (SUM). Afterwards, principal component analysis (PCoA) was performed using the scores of the seven FEs obtained using the first method. Subsequently, weights were created for each FE, based on each variable’s contribution variability obtained from the PCoA (SUM.W). Lastly, we applied a general score (GFS) to the animal’s ...

Facial-expression-based analysis has been widely applied as a pain coding system in horses. Herein, we aimed to identify pain in horses undergoing subcutaneously polylactide-based polymer implantation. The sham group was submitted only to surgical incision. The horses were filmed before and 24 and 48 h after implantation. Five statistical methods for evaluating their facial expressions (FEs) were tested. Primarily, three levels of scores (0, 1, and 2) were applied to the seven FEs (ear movements, eyebrow tension, orbicularis tension, dilated nostrils, eye opening, muzzle tension, and masticatory muscles tension). Subsequently, the scores of the seven FEs were added (SUM). Afterwards, principal component analysis (PCoA) was performed using the scores of the seven FEs obtained using the first method. Subsequently, weights were created for each FE, based on each variable’s contribution variability obtained from the PCoA (SUM.W). Lastly, we applied a general score (GFS) to the animal’s ...

Mastitis is the inflammatory reaction of the udder to invading pathogens, characterized by pathological changes in the mammary tissue, an increase in the number of somatic cells, physical, chemical and microbiological changes in the milk. We have investigated the relationship between somatic cells counts (SCC) and the concentration of antioxidant vitamins (retinol and α-tocopherol), the degree of oxidative degradation of lipids and caseins concentration. Increasing the number of somatic cells has the effect of decreasing concentration of vitamins studied. A normal SCC was correlated with the content of retinol 17.26 ± 0.86 respectively αtocopherol 63.6 ± 3.08 µg/ml. A SCC number greater than 500.000/ml determine a decrease in retinol concentration, the average value 13.87 ± 0.90 µg/ml and the decrease in α-tocopherol concentration, the average values 50.27 ± 1.27 µg/ml. Total protein content and the proportions of casein and whey protein in total protein undergo significant changes, even when sub clinical mastitis was only slight. Dropping antioxidants concentration is correlated with an increase in oxidation processes at the level of lipids, particularly fatty acids, as demonstrated by the increasing concentration malonildialdehyde (MDA-peroxidation final product).

The Quarter-Horses regularly participate in non-traditional sports that simulate activities on cattle ranches. In the vaquejada races, the horses run at about 30 km/h, reaching maximum heart rates of around 200 bpm, indicating the great physical effort during competitions and with that could impact welfare. To test the hypothesis that vaquejada horses can be evaluated for the quality of welfare through a combination of non-invasive and invasive methods, an experiment was developed that aimed to assess the quality of welfare through thermography and blood biomarkers before and after vaquejada races. Ten vaquejada horses, which were in regular competition, were submitted to the vaquejadas racing test were used. Thermography was performed in 14 regions of interest (head, neck and thorax) in the following phases: pre-race and +1, +4 and +24 hours of the races. Blood samples were collected in pre-race, immediately after and +1, +4 and +24 hours of the races. Results were submitted to ANOVA and Tukey's test, with p set at 5%. Thermography showed that local temperatures before the race and +24 hours after the races were similar (p > 0.05), as well as between +1 and +4 hours after the races (p > 0.05), which showed higher surface temperatures than the first group (pre-race and +24 hours) (p < 0.05). There was also an increase in red blood cells, hemoglobin and hematocrit immediately after the vaquejada races (p < 0.05) with a return to normality +1 hour after the races.

This research aimed to evaluate the effects of using either grain or commercial starter culture on the antioxidative capacity of cow and ewe milk kefirs. The antioxidant capacity of kefir samples during fermentation and 21 d of storage was assessed by using 3 assays: 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical cation decolorization; 2,2-diphenyl-1-picrylhydrazyl (DPPH •) radical scavenging activity assay; and Fe +3-reducing power (ferric reducing antioxidant power assay, FRAP). Vitamin E and β-carotene contents were also quantified. All kefir samples exhibited varying values for DPPH, ABTS, and FRAP assays depending on the starter culture and milk type. Vitamin E and β-carotene contents were similar in all kefir samples during storage. The results of this study suggest that milk type (cow or ewe) and culture type (kefir grains or commercial starter) were the significant parameters for the antioxidative activity of kefir.

This work presents the simple and low cost synthesis of a new tripodal ligand, in which three units of kojic acid are coupled to a tris(2-aminoethyl)amine (tren) backbone molecule. The protonation equilibria, together with the complex formation equilibria of this ligand with Fe 3+ , Al 3+ , Cu 2+ and Zn 2+ ions were studied. The complementary use of potentiometric, spectrophotometric and NMR techniques, and of Density Functional Theory (DFT) calculations, has allowed a thorough characterization of the different species involved in equilibrium. The stability of the formed complexes with Fe 3+ and Al 3+ are high enough to consider the new ligand for further studies for its clinical applications as a chelating agent. Biodistribution studies were carried out to assess the capacity the ligand for mobilization of gallium in 67 Ga-citrate injected mice. These studies demonstrated that this ligand efficiently chelates the radiometal in our animal model, which suggests that it can be a promising candidate as sequestering agent of iron and other hard trivalent metal ions. Furthermore, the good zinc complexation capacity appears as a stimulating result taking into a potential use of this new ligand in analytical chemistry as well as in agricultural and environmental applications.

The pathogenicity of group A Streptococcus (GAS) is mediated by direct bacterial invasivity and toxin-associated damage. Among the extracellular products, the exotoxin streptolysin O (SLO) is produced by almost all GAS strains. SLO is a pore forming toxin (PFT) hemolitically active and extremely toxic in vivo. Recent evidence suggests that human serum albumin (HSA), the most abundant protein in plasma, is a player in the innate immunity "orchestra." We previously demonstrated that HSA acts as a physiological buffer, partially neutralizing Clostridioides difficile toxins that reach the bloodstream after being produced in the colon. Here, we report the in vitro and ex vivo capability of HSA to neutralize the cytotoxic and hemolytic effects of SLO. HSA binds SLO with high affinity at a non-conventional site located in domain II, which was previously reported to interact also with C. difficile toxins. HSA:SLO recognition protects HEp-2 and A549 cells from cytotoxic effects and cell membrane permeabilization induced by SLO. Moreover, HSA inhibits the SLO-dependent hemolytic effect in red blood cells isolated from healthy human donors. The recognition of SLO by HSA may have a significant protective role in human serum and sustains the emerging hypothesis that HSA is an important constituent of the innate immunity system.

Ni/Al, Ni/Mn and Al/Mn co-doped monetite samples, as well as the undoped monetite, were prepared via wet chemical method. The structural characterization of the as-synthesized samples was done using X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) spectroscopy techniques. It is seen that all the samples are well-crystallized, except for Al/Mn co-doped monetite, and their phase compositions and crystal structures are affected significantly by kind of the as-mentioned co-dopants. Plate-like morphology, which is independent of the compositions of the samples, is observed for each sample. It is seen that all the as-investigated parameters can be controlled by kind of the co-additives.

Objective: Current concept distinguishes between cross intolerance (non-immune) and single or multiple hypersensitivity based (immune) adverse reactions of non-steroidal anti-inflammatory drugs (NSAID) due to their potential to inhibit cyclooxygenase (COX) isoenzymes (COX-1, COX-2). Recently we described a rapid IL-6 release assay using blood mononuclear cells of patients with various clinical forms of drug hypersensitivity. Here we present data of a comprehensive analysis of the IL-6 release test and the classical IgE immuno-assay for their sensitivity in cases with adverse reactions to NSAIDs grouped according to the new clinical classification. Methods: Total and specific serum IgE against 9 different HSA coupled-NSAIDs were determined by manual ELISA tests (55 cases) and compared to drug-specific release from preformed IL-6 pool of PBMCs of patients sensitized to the same NSAIDs after short (20 ') incubation of 4 standardized concentrations (51 cases and 9 controls) and IL-6 measurement from their cell free supernatants including positive and negative intraassay controls. Results: The ratio of cross intolerant to specific hypersensitive (HS) cases was higher in the IgE group (and total IgE too,) than in the IL-6 release tested ones. There was no difference, however, in the overall ratio of early and accelerated plus late onset adverse events based on individual histories. Nine NSAIDs were tested in both groups which represented all major COX-1 inhibitors. The positivity of validated test results was double within the IL-6 tested group (65.4% vs. 36.9%). In some cases non-drug components of NSAID formulations were responsible for the observed (mainly) anaphylactic reactions. Positive results in both groups were scattered amongst cross intolerant and single to multiple hypersensitive (HS) subgroups. To our knowledge no comprehensive analysis had been performed before either on clinical phenotypes dependent IgE immunoassays or on NSAID-induced "early" Tcell activation after those specified adverse events. Conclusion: Specific HS and multiple non cross-reactive NSAID sensitizations exceeded non-immune reactions in both in vitro tested groups. Some intolerant patients revealed detectable ASA antibodies of IgE type. Preformed IL-6 release by PBMC was more sensitive than specific IgE immunoassays as an in vitro diagnostic tool. The results indicate that checking of non-drug components should be considered in allergy workups. ASA in vivo provocations need further standardization.

The genotoxicity of anatase/rutile TiO 2 nanoparticles (TiO 2 NPs, NM105 at 3, 15 and 75 µg/cm 2) was assessed with the mammalian in-vitro Hypoxanthine guanine phosphoribosyl transferase (Hprt) gene mutation test in Chinese hamster lung (V79) fibroblasts after 24 h exposure. Two dispersion procedures giving different size distribution and dispersion stability were used to investigate whether the effects of TiO 2 NPs depend on the state of agglomeration. TiO 2 NPs were fully characterised in the previous European FP7 projects NanoTEST and NanoREG2. Uptake of TiO 2 NPs was measured by transmission electron microscopy (TEM). TiO 2 NPs were found in cytoplasmic vesicles, as well as close to the nucleus. The internalisation of TiO 2 NPs did not depend on the state of agglomeration and dispersion used. The cytotoxicity of TiO 2 NPs was measured by determining both the relative growth activity (RGA) and the plating efficiency (PE). There were no substantial effects of exposure time (24, 48 and 72 h), although a tendency to lower RGA at longer exposure was observed. No significant difference in PE values and no increases in the Hprt gene mutant frequency were found in exposed relative to unexposed cultures in spite of evidence of uptake of NPs by cells.

Copper (Cu)-doped calcium silicate nanoparticles were synthesized by a wet precipitation method as economical bone fracture filler. The aim was to improve the overall physicochemical properties, bioactivity, and biological performance of the bone fracture filler prepared herein. The synthesized nanoparticles were evaluated using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), and transmission electron microscopy (TEM). The bioactivity of the prepared nanoparticles was investigated after immersion in simulated body fluid (SBF) by means of inductively coupled plasma (ICP), SEM coupled with energy dispersive Xrays (EDX), and FTIR. The size and bioactivity of the prepared nanoparticles after 15 days of immersion in SBF was dependent on the Cu concentrations. The fracture healing ability of the fabricated nanoparticles on adult aged male Wistar rats was enhanced by the presence of copper. All the obtained results are of high relevance for fabricating improved Cudoped calcium silicate nanoparticles (50 nm) as low cost bone fracture filler. In addition, the in vivo study presented complete healing of the tibiae bone with normal architecture of bone tissue specifically calcium silicate nanoparticles doped with 3% and 5% Cu. Hence, the presence of copper is a promising tactic for improving the biological properties of calcium silicate. Therefore, the designed nanoparticles have huge potential for the treatment of bone fractures.

This study aims to evaluate the antioxidative activities of cow-milk kefir and goat-milk kefir. Antioxidative mechanisms, including radical-scavenging effects, ferrous-ion chelating ability, reducing power and antioxidant activity, were investigated herein. Kefirs demonstrated significantly greater scavenging effects upon 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide radicals, an inhibition effect upon linoleic-acid peroxidation, and more substantial reducing power, but reduced glutathione peroxidase (GSH-Px) activity than was the case for milks. There was no significant difference between milks and kefirs as regards ferrous-ion chelating ability and superoxide dismutase (SOD) activity. These findings have demonstrated that kefirs possess antioxidant activity, thereby suggesting that kefirs are potential candidates for the role of useful natural antioxidant supplements for the human diet.

Angiogenesis is essential to tumor progression, and the precise imaging of the angiogenic marker vascular endothelial growth factor receptor 2 (VEGFR-2) may provide an accurate evaluation for angiogenesis during a therapeutic response. With the use of molecular magnetic resonance imaging (mMRI), an in vitro cell assay indicated significantly decreased T1 relaxation values when tumor endothelial cells (TEC), which positively expressed VEGFR-2 (Western blot), were in the presence of the VEGFR-2 probe compared to TEC alone (P &lt; 0.001). For in vivo mMRI evaluations, we assessed VEGFR-2 levels in untreated and OKN-007-treated GL261 mouse gliomas. Regarding treatment response, OKN-007 was also able to significantly decrease tumor volumes (P &lt; 0.01) and increase survival (P…

In this study, biocompatible titanium-niobium (Ti-Nb) alloys were fabricated by using powder metallurgy methods. Physical, morphological, thermal, and mechanical analyses were performed and their in vivo compatibility was evaluated. Besides α, β, and α″ martensitic phases, α+β Widmanstätten phase due to increasing sintering temperature was seen in the microstructure of the alloys. Phase transformation temperatures of the samples decreased as Nb content increased. The ratio of Nb in the samples affected their mechanical properties. No toxic effect was observed on implanted sites. This study shows that Ti-Nb alloys can be potentially used for orthopedic applications without any toxic effects.

The Quarter-Horses regularly participate in non-traditional sports that simulate activities on cattle ranches. In the vaquejada races, the horses run at about 30 km/h, reaching maximum heart rates of around 200 bpm, indicating the great physical effort during competitions and with that could impact welfare. To test the hypothesis that vaquejada horses can be evaluated for the quality of welfare through a combination of non-invasive and invasive methods, an experiment was developed that aimed to assess the quality of welfare through thermography and blood biomarkers before and after vaquejada races. Ten vaquejada horses, which were in regular competition, were submitted to the vaquejadas racing test were used. Thermography was performed in 14 regions of interest (head, neck and thorax) in the following phases: pre-race and +1, +4 and +24 hours of the races. Blood samples were collected in pre-race, immediately after and +1, +4 and +24 hours of the races. Results were submitted to ANOVA and Tukey's test, with p set at 5%. Thermography showed that local temperatures before the race and +24 hours after the races were similar (p > 0.05), as well as between +1 and +4 hours after the races (p > 0.05), which showed higher surface temperatures than the first group (pre-race and +24 hours) (p < 0.05). There was also an increase in red blood cells, hemoglobin and hematocrit immediately after the vaquejada races (p < 0.05) with a return to normality +1 hour after the races.

Different soybean products are considered as traditional functional food among the Mongolian population in Northeast India. In the present study, the effect of different flavours (mango, orange, vanilla and white rose), inoculation rates of Lactobacillus rhamnosus K4E (KX950834) and Lactobacillus helveticus K14 (KU644578), and mass fractions of skimmed milk and sugar on the acceptability of soy yoghurts was studied. Physicochemical (pH, titratable acidity) and microbial analyses (total bacterial, total coliform, yeast and mould count) were conducted, and organoleptic (aroma, taste, colour, mouthfeel, texture and overall acceptability) and biofunctional properties (angiotensin-converting enzyme (ACE) inhibitory activity, antioxidant and antimicrobial activities and biotransformation of isoflavones) were evaluated during storage up to 10 days under refrigeration conditions (6-8 °C). Panellists preferred white rose soy yoghurt more than other flavours. The pH was from 5.65 to 4.20, the titratable acidity (expressed as mass fraction of lactic acid) was from 0.33 to 0.51 % and total Lactobacillus count ranged from 6.81 to 8.69 log CFU/mL during storage. The ACE inhibitory activity increased from 21.17 % on day 0 to 81.03 % on day 5, followed by a decrease of the activity after 10 days (38.85 %). The antioxidant activity was the highest on day 5 (87 %). White rose soy yoghurt had the highest antimicrobial activity against Listeria monocytogenes, followed by Bacillus subtilis, Staphylococcus aureus, Salmonella typhi and Escherichia coli. RP-HPLC analysis showed that after 18 h, the production of soy isoflavone aglycones genistein and daidzein in yoghurt was 87.3 and 58.4 %, respectively.

During conventional fermentation of milk into yogurt and kefir, changes of antioxidant capacities (DPPH radical scavenging, ferrous chelating, reducing power, hydrogen peroxide/ H2O2 scavenging activity) and total phenolic content were investigated. Both products showed greater phenolic content, DPPH radical scavenging, and reducing power activities than was the case of their unfermented milk forms. But they were unable to scavenge H2O2. Chelating ability of milk increased by yogurt fermentation but decreased by kefir fermentation. In general, antioxidative capacity of milk improved throughout conventional yogurt and kefir fermentations and ripening periods (except chelating ability for kefir and H2O2 scavenging). This study is valuable since it evaluates the antioxidant power of yogurt and kefir produced by traditional fermentation with 5 different antioxidative approaches. In face of rapidly rising consumer demand for safe, healthy, functional but natural foods, revealed results might strengthen importance of conventional fermentation process and highlight the antioxidative contributions of mentioned products.

Objective: A triphasic bone graft block composed of gypsum, brushite, and monetite is expected to be better for regenerating bone than a gypsum-hydroxyapatite-tricalcium phosphate block. Therefore, the aim of this study was to fabricate and evaluate the mechanical properties of a newly developed triphasic block composed of gypsum, brushite, and monetite. Materials and method: Triphasic blocks were prepared by mixing calcium sulfate hemihydrate, brushite, and monetite powders with distilled water at a powder-to-liquid ratio of 0.5. The content of calcium sulfate hemihydrate was fixed at 50%, and the contents of brushite and monetite powders were varied. After molding and setting, the obtained blocks were characterized, and their mechanical properties were evaluated. Results: The triphasic blocks were prepared and could maintain their shape without collapsing. The XRD characterization of the obtained triphasic blocks showed that only three phases existed in the block. Calcium sulfate hemihydrate was transformed into its dihydrate form and provided mechanical strength to the block through a setting mechanism. The transformation of calcium sulfate hemihydrate into its dihydrate crystals formed an interlocked structure that was disrupted in triphasic blocks, as observed in SEM images. The disruption of the interlocked structure resulted

Background and Objective: In this study, nine Lactobacillus kefiri and one Lactobacillus rhamnosus isolates with kefir grain origin have been demonstrated to include potentials as probiotics. The aim of this study was to investigate ability of the isolates to produce compounds with α-glucosidase inhibitory and antioxidant activities and identify peptides with MW of ≤ 3 kDa in cell-free supernatants. Material and Methods: All isolates were cultured in de Man, Rogosa and Sharpe broth media at 37 oC for 24 h. Assessment of α-glucosidase inhibitory and antioxidant activities was carried out on cell-free supernatants. Assessment of optimum incubation time was carried out on two isolates with the highest α-glucosidase inhibitory and antioxidant activities. The two isolates were used to ferment reconstituted skim milk. Cell-free supernatant of the fermented skim milk was fractionated using filters of 10 and 3 kDa. Then, peptides in fractions of ≤ 3 kDa were identified. Results and Conclusi...

The present study describes the effects of dopants of Mn and/or Ni on the structural, magnetic and in vivo/in vitro performances of β-tricalcium phosphate (β-TCP) bioceramics synthesized by a sol-gel method. Both types of dopants caused significant effects on the structure-related parameters of β-TCP, including the crystallite dimensions and lattice parameters. The magnetic properties of as-prepared samples were found to be affected by the type and amount of the dopant(s). The in vitro antimicrobial activities of doped β-TCPs against American Type Culture Collection (ATCC) reference bacterial and fungal strains were evaluated. Among them, only 1.35Ni-TCP displayed an effective antimicrobial activity against Gram-positive bacteria (50 μg/mL Minimum Inhibitory Concentration (MIC) range), while other types of doped β-TCPs did not possess with such effect. The in vivo tests showed that each formulation could be a good nominate for bone healing and bone treatment.

Reducing the salt content in food is an important nutritional strategy for decreasing the risk of diet-related diseases. This strategy is particularly effective when applied to highly appreciated food having good nutritional characteristics, if it does not impact either upon sensory or nutritional properties of the final product. This work aimed at evaluating if the reduction of salt content by decreasing the brine soaking time modifies fatty acid and protein bioaccessibility and bioactive peptide formation in a 30-month-ripened Parmigiano Reggiano cheese (PRC). Hence, conventional and hyposodic PRC underwent in vitro static gastrointestinal digestion, and fatty acid and protein bioaccessibility were assessed. The release of peptide sequences during digestion was followed by LC–HRMS, and bioactive peptides were identified using a bioinformatic approach. At the end of digestion, fatty acid and protein bioaccessibility were similar in conventional and hyposodic PRC, but most of the bi...

Soy milk, a derivative of soybean, is an alternative to dairy beverage, but its acceptability is limited worldwide due to unpleasant beany flavour. Fermentation may, however, improve the nutritional and sensory values of soy milk. Thus, post-fermentation improvement in functional attributes of soy milk were investigated via antioxidant, ACE inhibitory, and antimicrobial activities in this study using five test and one control strains of lactobacilli. Results indicated that soy milk fermented by Lactobacillus rhamnosus strain C25 (LR C25) effectively scavenged more than 60% of the ABTS, DPPH and Hydroxyl radicals generated in in vitro models. Moreover, all the strains showed significantly higher (p < 0.05) antioxidant activity as compared to unfermented soy milk in all three assays performed. Further, Lactobacillus plantarum strain C6 (LP C6) fermented soy milk displayed significantly higher (p < 0.05) percent ACE inhibitory activity (68.40 ± 0.93%) as compared to other tested Lactobacillus isolates, reference strain and unfermented soy milk. It was also observed that LP C6 strongly inhibited the growth of indicator strain of E. coli in the agar well diffusion assay. These strains can therefore be further explored in the preparation of beneficial soy foods and bioactive food supplements for wellbeing.

Gliomas are the most common brain tumors, with diverse biological behaviour. Glioblastoma (GBM), the most aggressive and with the worst prognosis, is characterized by an intense and aberrant angiogenesis, which distinguishes it from low-grade gliomas (LGGs) and benign expansive lesions, as meningiomas (MNGs). With increasing evidence for the importance of vascularization in tumor biology, we focused on the isolation and characterization of endothelial cells (ECs) from primary GBMs, LGGs and MNGs. Gene expression analysis by Real-Time PCR, immunofluorescence and flow cytometry analysis, tube-like structures formation and vascular permeability assays were performed. Our results showed a higher efficiency of ECs to form a complex vascular architecture, as well as a greater impairment of a brain blood barrier model, and an overexpression of pro-angiogenic mediators in GBM than in LGG and MNG. Furthermore, administration of temozolomide, bevacizumab, and sunitinib triggered a different proliferative, apoptotic and angiogenic response, in a dose and time-dependent manner. An increased resistance to temozolomide was observed in T98G cells co-cultured in GBM-EC conditioned media. Therefore, we developed a novel platform to reproduce tumor vascularization as "disease in a dish", which allows us to perform screening of sensitivity/resistance to drugs, in order to optimize targeted approaches to GBM therapy. Central Nervous System (CNS) tumors are among the most complex human cancers, with a prognosis closely related to histological classification, location, genetic background and specific microenvironment of the tumor. Brain tumors present anatomically similar features, but they differ significantly for morphology, aetiology, site of origin, molecular biology and clinical progression 1. In adults, about 50% of all CNS tumors are malignant, whereas in paediatric patients, more than 75% 2. The ability of tumor mass to grow, progress and infiltrate into surrounding tissue, compromising cognitive and motor skills, is due to the intense and aberrant angiogenesis, responsible for the formation of new blood vessels, which support tumor development, providing nutrition, oxygen and energy 3 , in a mechanism well known as neovascularization. Angiogenesis is regulated by a number of both stimulating and inhibiting angiogenic factors acting on endothelial cells (ECs) 4. Often, concentration of pro-angiogenic mediators increase progressively with tumor grade and is linked with the clinical outcome 5. In

Health benefits of probiotics and production of inhibitors of angiotensin converting enzyme (ACE) released during milk fermentation are well known. That is why in this investigation the proteolytic profile and ACE inhibitory capacity of peptide fractions from protein hydrolysis of milk during fermentation processes was analyzed. Milk fermentation was carried out inoculating 106 CFU of L. rhamnosus GG, S. thermophilus SY-102 and with both bacteria. The proteolytic profile was determined using: TNBS, SDS-PAGE and SEC-HPLC techniques. In vitro ACE inhibition capacity was measured. The pH of 4.5 was reached at 56 h when the milk was fermented with L. rhamnosus, at 12 h with S. thermophillus and at 41 h in the co-culture. Production of free amino groups corresponded with the profile of low molecular weight peptides observed by SDS-PAGE and SEC-HPLC. Co-culture fermentation showed both the highest concentration of low molecular weight peptides and the ACE inhibitory activity (&gt;80%). Re...

Abstract
Inflammation is a regulated response to injury or infection that protects and repairs the body. When an inflammatory response is unregulated, hyper-inflammation can lead to organ damage and endothelium dysfunction. Recently, SARS-CoV-2 infections have been the cause of severe pneumonia, cardiovascular complications, and organ failures. Severe infections are accompanied by pulmonary hyper-inflammation and induced vascular endothelium damage. Protein nanoparticles can be modified to target and deliver anti-inflammatory cargo to the source of inflammation which is primarily among vascular endothelium cells. Specifically, this thesis explores the fabrication and optimization of protein nanoparticles with conjugated antibodies to specifically target endothelial cells. Protein nanoparticle targeting and uptake were confirmed in the cytoplasm of cultured Human Umbilical Vascular Endothelial Cells (HUVEC) using confocal microscopy and flow cytometry. Lastly, the biodistribution of these nanoparticles was assessed in healthy mice to understand the targeting of protein nanoparticles conjugated with targeting antibodies in vivo. Overall, this thesis demonstrates the impact that conjugated antibodies have on enhancing intracellular delivery and the biodistribution of these nanoparticles in vivo.

This study aims to evaluate the antioxidative activities of cow-milk kefir and goat-milk kefir. Antioxidative mechanisms, including radical-scavenging effects, ferrous-ion chelating ability, reducing power and antioxidant activity, were investigated herein. Kefirs demonstrated significantly greater scavenging effects upon 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide radicals, an inhibition effect upon linoleic-acid peroxidation, and more substantial reducing power, but reduced glutathione peroxidase (GSH-Px) activity than was the case for milks. There was no significant difference between milks and kefirs as regards ferrous-ion chelating ability and superoxide dismutase (SOD) activity. These findings have demonstrated that kefirs possess antioxidant activity, thereby suggesting that kefirs are potential candidates for the role of useful natural antioxidant supplements for the human diet.

Lactobacillus casei ATCC 393 is an important probiotic strain widely known in dairy technology. However, its capability to produce bioactive peptides from milk proteins has not been studied. The viability of the Lb. casei ATCC 393 strain and some physicochemical properties in fermented skim milk throughout storage for 21 days at 4 °C was evaluated; biological activity, i.e., antioxidant, angiotensin converting enzyme inhibitory and anticancer activities of water soluble extract and its filtrate (˂ 2 kDa; F1) were determined. Lb. casei counts remained over 9 log cfu g-1 during the storage period in fermented milk. These bioactivities were increased significantly (P < 0.01) during storage. F1 of fermented milk after three weeks of storage showed the highest bioactivity impact. De novo sequencing assay for peptide identification was applied to the mass spectrum of F1. The promising capability of Lb. casei ATCC 393 to release bioactive peptides from milk proteins was demonstrated.

Heat treatments may cause some chemical and physicochemical changes in milk, although milk is a heat-stable system. Heat treatments can cause different changes in different types of milk. This study aimed to compare the effects of pasteurization and boiling on goat and cow milk's macromolecular contents, glutathione levels, and superoxide dismutase activities. The protein level of both types of milk decreased with the pasteurization process, and boiling also reduced the protein level of goat milk. Both heat treatments reduced superoxide dismutase activity and glutathione levels in both types of milk. While the boiling process did not change the cow's milk lactose level, it increased the goat milk lactose level. It was determined that pasteurization reduced the lactose level in both types of milk. Pasteurization did not change the fat level in cow milk but decreased the fat level in goat milk. In conclusion, cow milk was less affected by these heat treatments, which can be attributed to having large fat globules, high lactose concentration, and high heat resistance protein content compared to goat milk.

Lantibiotics continue to offer an untapped pipeline for the development of novel antibiotics. We report here the discovery of a novel lantibiotic for the treatment of C. difficile infection (CDI). The leads were selected from a library of over 300 multiple substitution variants of the lantibiotic Mutacin 1140 (MU1140). Top performers were selected based on testing for superior potency, solubility, manufacturability, and physicochemical and/or metabolic stability in biologically-relevant systems. The best performers in vitro were further evaluated orally in the Golden Syrian hamster model of CDAD. In vivo testing ultimately identified OG716 as the lead compound, which conferred 100% survival and no relapse at 3 weeks post infection. MU1140-derived variants are particularly attractive for further clinical development considering their novel mechanism of action.

Kefir, a traditional fermented food, has numerous health benefits due to its unique chemical composition, which is reflected in its excellent nutritional value. Physicochemical and microbial composition of kefir obtained from fermented milk are influenced by the type of the milk, grain to milk ratio, time and temperature of fermentation, and storage conditions. It is crucial that kefir characteristics are maintained during storage since continuous metabolic activities of residual kefir microbiota may occur. This study aimed to examine the nutritional profile of kefir produced in traditional in use conditions by fermentation of ultra-high temperature pasteurized (UHT) semi-skimmed cow milk using argentinean kefir grains and compare the stability and nutritional compliance of freshly made and refrigerated kefir. Results indicate that kefir produced under home use conditions maintains the expected characteristics with respect to the physicochemical parameters and composition, both afte...

This work presents the simple and low cost synthesis of a new tripodal ligand, in which three units of kojic acid are coupled to a tris(2-aminoethyl)amine (tren) backbone molecule. The protonation equilibria, together with the complex formation equilibria of this ligand with Fe 3+ , Al 3+ , Cu 2+ and Zn 2+ ions were studied. The complementary use of potentiometric, spectrophotometric and NMR techniques, and of Density Functional Theory (DFT) calculations, has allowed a thorough characterization of the different species involved in equilibrium. The stability of the formed complexes with Fe 3+ and Al 3+ are high enough to consider the new ligand for further studies for its clinical applications as a chelating agent. Biodistribution studies were carried out to assess the capacity the ligand for mobilization of gallium in 67 Ga-citrate injected mice. These studies demonstrated that this ligand efficiently chelates the radiometal in our animal model, which suggests that it can be a promising candidate as sequestering agent of iron and other hard trivalent metal ions. Furthermore, the good zinc complexation capacity appears as a stimulating result taking into a potential use of this new ligand in analytical chemistry as well as in agricultural and environmental applications.

Contesto di riferimento: I molteplici cambiamenti che hanno caratterizzato lo sviluppo economico sociale negli ultimi decenni hanno generato profondi mutamenti nei comportamenti di consumo, coinvolgendo, in particolare le dinamiche del consumo alimentare. In tutte le economie sviluppate i mutamenti sono stati e sono rapidi e significativi. Ancora di più nelle economie emergenti dove il cambiamento è stato straordinariamente veloce ed ha portato addirittura a mettere in crisi l'equilibrio tra risorse produttive e consumi alimentari. Negli ultimi anni si è verificata la rapida crescita di alcuni mercati quale quello dei prodotti tipici, dei prodotti biologici, dei prodotti funzionali e quello dei prodotti ad alto valore aggiunto e contenuto di servizio. La cattiva dieta (nutrizione) e il fumo di tabacco sono risultati tra i principali fattori di rischio ambientale per malattie cardiovascolari, cancro e diabete. Si sono moltiplicate le raccomandazioni dietetiche e si è cercato di orientare la popolazione su stili di vita più “salutari”: controllo del peso, controllo del livello di colesterolo, controllo della pressione arteriosa.
Scopo: Il progetto di ricerca e sviluppo ha come obiettivo l’innovazione di prodotto e di processo per la realizzazione di snack, merendine e spezza fame ad elevato valore funzionale, salutistico e antiossidante con l’ausilio di mild technologies. Le peculiarità su cui si intende concentrare lo studio, con evidenti ripercussioni sul processo di produzione, sono il ridotto contenuto calorico dei prodotti, l’uso di materie prime con alto contenuto di molecole bioattive, nuove forme distributive. L’intento è, dunque, sviluppare nuove formulazioni in cui ingredienti di elevato valore nutrizionale siano utilizzati in alimenti innovativi presenti nella dieta al fine di ottenere un intake significativo di componenti funzionali.
Materiali e metodi: Al fine di perseguire l’obiettivo finale, verrà condotto prima uno screening delle materie prime da utilizzare nelle formulazioni e successivamente verranno analizzate le specifiche dei nuovi prodotti funzionali e delle tecnologie da sviluppare. I prodotti nutraceutici così ottenuti saranno sottoposti ad una valutazione clinica secondo le linee EFSA e GCP e successivamente sarà validatala loro efficacia salutistica. Infine, sarà valutata la sostenibilità economica ed ambientale delle innovazioni e analisi delle preferenze del consumatore.

Bu çalışmada pastörize, yarım yağlı UHT ve UHT sütlerin oksidan ve antioksidan parametrelerinin değerlendirilmesi için malondialdehit (MDA), vitamin (Vit) C, total protein (TP), redükte glutatyon (rGSH) ve katalaz (CAT) belirlenmesi ve bu parametreler arasındaki korelasyonların ortaya çıkarılması amaçlandı. Gereç ve Yöntem: Hatay ilinde farklı marketlerden farklı markalara ait 30 adet süt örneği toplandı. Süt MDA, Vit C, TP, rGSH düzeyleri ve CAT aktiviteleri spektrofotometrik yöntemlerle belirlendi. Bulgular: En düşük MDA düzeyi yarım yağlı UHT sütte belirlendi (p < 0,05). Pastörize sütün MDA düzeyi yarım yağlı UHT sütten daha yüksekti (p < 0,05). Pastörize sütlerde Vit C (p < 0,001) ve TP (p < 0,01) düzeyleri diğer sütlere göre daha yüksek bulundu. Pastörize ve UHT sütlerin rGSH düzeyleri, yarım yağlı UHT sütlere göre daha yüksekti (p < 0,05). UHT yarım yağlı süt CAT aktivitesi diğer sütlere göre oldukça yüksek bulundu (p < 0,05). Korelasyonlar açısından; pastörize süt MDA seviyesi, Vit C seviyesi ile pozitif korelasyon gösterdi (r = 0,743, p < 0,01). Yarım yağlı UHT süt TP seviyesi, CAT aktivitesi ile pozitif korelasyon gösterdi (r=0,673, p < 0,05). Öneri: Sonuç olarak sunulan çalışmada, pastörize, yarım yağlı UHT ve UHT sütün oksidan (MDA), antioksidan parametreleri (Vit C, rGSH, and CAT), ve TP düzeyleri değerlendirildi ve tartışıldı. Bu çalışmada araştırılan değerler açısından, pastörize süt tüketiminin diğer ticari sütlere göre daha sağlıklı olabileceği önerilmektedir.

Aim: The phytochemical screening, antibacterial activities and in vivo toxicity of extracts of the leaves of scent leaf (Ocimum gratissimum L.) were investigated. Methods: All the analyses were carried out using standard scientific procedures using Soxhlet extraction, well-diffusion agar antimicrobial testing and in vivo acute toxicity testing. Results: The phytochemical analysis according to standard screening tests using conventional protocols revealed the presence of anthraquinone, saponins, tannins, terpenoids and alkaloids, which were detected in methanol extract analyzed. But, flavonoids, glycosides, phlobatannins and steroids were not detected in the methanol extracts analyzed. While only flavonoids were detected in chloroform extract. All other phytochemicals were absent. The extract fractions generally exhibited slight antibacterial activities on Staphylococcus aureus, Bacillus subtilis, Salmonella Typhi and Escherichia coli. But, the extracts showed no effect against Candi...

Lipid oxidation is one of the key determinants of the quality of iron-fortified dairy products. In this study, using starter cultures possessing a high ability to release metal-binding peptides during fermentation and storage was proposed to address the quality problems related to fortification of fermented milk with iron. For this purpose, a combination of commercial yogurt starter (CS) with Lactobacillus plantarum B-4496 (LP) or Lactobacillus paracasei subsp . paracasei B-4560(LPC)wasused to ferment camel milk, which was fortified with ferrous sulphate at a level of 40 mg iron/kg milk. Fermented milk was stored at 4±1°C for 14 days. Proteolysis degree and thiobarbituric acid (TBA) value were evaluated for all treatments, while iron chelating activity (ICA) and fatty acids composition were determined for the iron-free treatments. The results revealed that starter culture combinations had a significant influence (p˂0.05) on all studied parameters. Samples containing CS-LPC showed th...

The implementation of &lt;em&gt;in vivo&lt;/em&gt; fluorescence imaging as a reliable diagnostic imaging modality at the clinical level is still far from reality. A lot of work remains ahead to provide medical practitioners with solid proof of the potential advantages of this imaging technique over well-established ones such as magnetic resonance imaging or nuclear imaging. To do so, one of the key objectives is to improve the optical performance of dedicated contrast agents, thus improving the resolution and penetration depth achievable with &lt;em&gt;in vivo&lt;/em&gt; imaging. In the present study, we move along this direction and report on the use of AgInSe&lt;sub&gt;2&lt;/sub&gt; nanoparticle-based contrast agents (nanocapsules) for fluorescence imaging. The use of a Ag&lt;sub&gt;2&lt;/sub&gt;Se seeds-mediated synthesis method allows stabilizing an uncommon orthorhombic crystal structure. This structure endows the material with emission in the second biological window (1000-140...