The majority of dry pet food currently on the market is produced using fresh meats (FMs) and especially meat meals (MMs) as the main protein source. The transport and storage conditions of the raw materials, together with thermal and mechanical treatments in the case of MMs, may result in undesirable alterations of food products and their protein content. This study was conducted to analyze the protein component of three different kinds of raw materials used for dry pet food production, i.e., chicken, pork, and salmon. The quantitative analysis of the protein component was determined using the traditional Kjeldahl method and near-infrared (NIR) spectroscopy, and an alternative method, i.e., the Bradford assay, while the qualitative analysis was performed through SDS-PAGE, followed by Coomassie Blue staining. The amino acid (AA) profile was also evaluated by quadrupole time-of-flight liquid chromatography/mass spectrometry (Q-TOF LC/MS). In addition, the digestibility was tested through in vitro gastric and small intestine digestion simulation. Statistical analysis was performed by the Student’s t-test, and data are reported as mean ± SEM, n = 10 (p < 0.05). The results showed that the MMs are lower in quality compared to FMs, both in terms of protein bioavailability and digestibility, having a lower soluble protein (SP) content (chicken MM = 8.6 g SP/100 g dry sample; pork MM = 6.2 g SP/100 g dry sample; salmon MM = 7.9 g SP/100 g dry sample) compared to FMs (chicken FM = 14.6 g SP/100 g dry sample; pork FM = 15.1 g SP/100 g dry sample; salmon FM = 13.7 g SP/100 g dry sample). FMs appear, therefore, to be higher-quality ingredients for pet food production. Moreover, the Bradford assay proved to be a quick and simple method to better estimate protein bioavailability in the raw materials used for dry pet food production, thanks to its correlation with the in vitro digestibility.
In this study, we report evidence for emissive aggregates of push−pull phenothiazine and phenothiazine dioxide derivatives produced both in water dispersions and in the solid state, highly promising for applications in both biology and material science. An insightful investigation of the aggregation-induced emission (AIE) mechanism was carried out via time-resolved spectroscopies, with nanosecond and femtosecond temporal resolution, coupled with advanced data analysis. Our steady-state and timeresolved spectroscopic results unambiguously show that a significant AIE behavior is activated by the restriction of intramolecular rotations for the phenothiazine derivatives. Investigation of the nonlinear optical properties also revealed that aggregates exhibit notable emission upon two-photon excitation. In particular, the phenothiazine dioxide-based aggregates exhibit remarkable fluorescence efficiencies and large two-photon absorption cross sections, as well as the capability to be internalized in HeLa cells exerting no cytotoxic effect. These aggregate species thus prove to be promising as novel fluorophores for bioimaging.
Diseases affecting the central nervous system (CNS) are among the most disabling and the most difficult to cure due to the presence of the blood–brain barrier (BBB) which represents an impediment from a therapeutic and diagnostic point of view as it limits the entry of most drugs. The use of biocompatible polymer nanoparticles (NPs) as vehicles for targeted drug delivery to the brain arouses increasing interest. However, the route of administration of these vectors remains critical as the drug must be delivered without being degraded to achieve a therapeutic effect. An innovative approach for the administration of drugs to the brain using polymeric carriers is represented by the nose-to-brain (NtB) route which involves the administration of the therapeutic molecule through the neuro-olfactory epithelium of the nasal mucosa. Nasal administration is a non-invasive approach that allows the rapid transport of the drug directly to the brain and minimizes its systemic exposure. To date, many studies involve the use of polymer NPs for the NtB transport of drugs to the brain for the treatment of a whole series of disabling neurological diseases for which, as of today, there is no cure. In this review, various types of biodegradable polymer NPs for drug delivery to the brain through the NtB route are discussed and particular attention is devoted to the treatment of neurological diseases such as Glioblastoma and neurodegenerative diseases.
β-d-N-acetyl-hexosaminidase (Hex, EC 3.2.1.52) is an acid hydrolase that catalyzes the cleavage of the β-1,4 bond in N-acetyl-d-galactosamine (Gal-NAc) and N-acetyl-d-glucosamine (Glc-NAc) from the non-reducing end of oligosaccharides and glycoconjugates. It is widely expressed in both the prokaryotic and eukaryotic world, where it performs multiple and important functions. Hex has antifungal activity in plants, is capable of degrading many biological substrates, and can play an important role in the biomedical field for the treatment of Tay-Sachs and Sandhoff diseases. With the aim being able to obtain a device with a stable enzyme, a method of covalent immobilization on polylactic acid (PLA) films was developed for the A isoform of the β-d-N-acetyl-hexosaminidase enzyme (HexA), produced in a recombinant way from Human Embryonic Kidney-293 (HEK-293) cells and suitably purified. An in-depth biochemical characterization of the immobilized enzyme was carried out, evaluating the optimal temperature, thermal stability, pH parameters, and Km value. Moreover, the stability of the enzymatic activity over time was assessed. The results obtained showed an improvement in terms of kinetic parameters and stability to heat for the enzyme following immobilization and the presence of HexA in two distinct immobilized forms, with an unexpected ability for one of them to maintain its functionality for a long period of time (over a year). The stability and functionality of the enzyme in its immobilized form are therefore extremely promising for potential biotechnological and biomedical applications.
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