Critical Assessment of a Structure-Based Screening Campaign for IDO1 Inhibitors: Tips and Pitfalls

Over the last two decades, indoleamine 2,3-dioxygenase 1 (IDO1) has attracted wide interest as a key player in immune regulation, fostering the design and development of small molecule inhibitors to restore immune response in tumor immunity. In this framework, biochemical, structural, and pharmacological studies have unveiled peculiar structural plasticity of IDO1, with different conformations and functional states that are coupled to fine regulation of its catalytic activity and non-enzymic functions. The large plasticity of IDO1 may affect its ligand recognition process, generating bias in structure-based drug design campaigns. In this work, we report a screening campaign of a fragment library of compounds, grounding on the use of three distinct conformations of IDO1 that recapitulate its structural plasticity to some extent. Results are instrumental to discuss tips and pitfalls that, due to the large plasticity of the enzyme, may influence the identification of novel and differentiated chemical scaffolds of IDO1 ligands in structure-based screening campaigns.     

Toxic Effects on Thyroid Gland of Male Adult Lizards (Podarcis Siculus) in Contact with PolyChlorinated Biphenyls (PCBs)-Contaminated Soil

Skin exposure is considered a potentially significant but little-studied pathway for PolyChlorinated Biphenyls uptake in terrestrial reptiles. In this study, a native Italian lizard, Podarcis siculus, was exposed to PCBs-contaminated soil for 120 days. Tissues distribution of PCBs, thyroid hormone levels, and thyroid histo-physiopathology were examined. The accumulation of PCBs in skin, plasma, liver, kidney, and brain were highest at 120 days. The alteration of triiodothyronine (T₃) and thyroxine (T₄) levels after different concentrations and times to exposure of PCBs was accompanied by the changes in the hormones involved in the hypothalamus-pituitary-thyroid (HPT) axis, namely Thyrotropin Releasing Hormone (TRH) and Thyroid Stimulating Hormone (TSH). Moreover, hepatic levels of deiodinase II (5′ORDII) and content of T₃ were positively correlated to exposure to PCBs. These results indicated that in lizards, PCBs exposure through the skin has the potential to disrupt the thyroid endocrine system. Overall, the observed results indicate that PCBs could be associated with changes in thyroid homeostasis in these reptiles, through direct interactions with the metabolism of T₄ and T₃ through the HPT axis or indirect interactions with peripheral deiodination.     

MicroRNA Signatures in the Upper Urinary Tract Urothelial Carcinoma Scenario: Ready for the Game Changer?

Upper urinary tract urothelial carcinoma (UTUC) represents a minor subgroup of malignancies arising in the urothelium of the renal pelvis or ureter. The estimated annual incidence is around 2 cases per 100,000 people, with a mean age at diagnosis of 73 years. UTUC is more frequently diagnosed in an invasive or metastatic stage. However, even though the incidence of UTUC is not high, UTUC tends to be aggressive and rapidly progressing with a poor prognosis in some patients. A significant challenge in UTUC is ensuring accurate and timely diagnosis, which is complicated by the non-specific nature of symptoms seen at the onset of disease. Moreover, there is a lack of biomarkers capable of identifying the early presence of the malignancy and guide-tailored medical treatment. However, the growing understanding of the molecular biology underlying UTUC has led to the discovery of promising new biomarkers. Among these biomarkers, there is a class of small non-coding RNA biomarkers known as microRNAs (miRNAs) that are particularly promising. In this review, we will analyze the main characteristics of UTUC and focus on microRNAs as possible novel tools that could enter clinical practice in order to optimize the current diagnostic and prognostic algorithm.     

A capacitive technique to assess water content in extra virgin olive oils

The present research investigated the correlations between capacitance and water content of extra virgin olive oils (EVOO). A commercial capacitor probe for radio applications and an LCR meter were used for electric tests in the frequency range from 500 Hz to 512 kHz. Seventeen samples of different EVOO with a moisture content ranging from 178 to 1321 mg/kg oil were selected for study. To assess the influence of moisture only, the oil with the maximum water content was filtered down to 288 mg/kg oil and five samples with intermediate water contents were prepared and submitted to electrical measurements. Subsequently, the capacitance of all 17 EVOO samples was measured at selected frequencies.     

Residues in the Distal Heme Pocket of Arabidopsis Non-Symbiotic Hemoglobins: Implication for Nitrite Reductase Activity

It is well-established that plant hemoglobins (Hbs) are involved in nitric oxide (NO) metabolism via NO dioxygenase and/or nitrite reductase activity. The ferrous-deoxy Arabidopsis Hb1 and Hb2 (AHb1 and AHb2) have been shown to reduce nitrite to NO under hypoxia. Here, to test the hypothesis that a six- to five-coordinate heme iron transition might mediate the control of the nitrite reduction rate, we examined distal pocket mutants of AHb1 and AHb2 for nitrite reductase activity, NO production and spectroscopic features. Absorption spectra of AHbs distal histidine mutants showed that AHb1 mutant (H69L) is a stable pentacoordinate high-spin species in both ferrous and ferric states, whereas heme iron in AHb2 mutant (H66L) is hexacoordinated low-spin with Lys69 as the sixth ligand. The bimolecular rate constants for nitrite reduction to NO were 13.3 ± 0.40, 7.3 ± 0.5, 10.6 ± 0.8 and 171.90 ± 9.00 M⁻¹·s⁻¹ for AHb1, AHb2, AHb1 H69L and AHb2 H66L, respectively, at pH 7.4 and 25 °C. Consistent with the reductase activity, the amount of NO detected by chemiluminescence was significantly higher in the AHb2 H66L mutant. Our data indicate that nitrite reductase activity is determined not only by heme coordination, but also by a unique distal heme pocket in each AHb.     

A Pilot Study for Identification of <Emphasis Type="Italic">Salmonella</Emphasis> in Food Processing Plants by Real-Time PCR Screening

Salmonella remains a major public health concern worldwide. Microbiological methods are the gold standard for Salmonella detection. These methods are highly specific, but their sensitivity is variable. Moreover, they are lengthy, labour intensive and not always consistent with the speed of food manufacturing processes. Thus, in the food industry, there is the need for more rapid, sensitive and accurate detection methods. The purpose of this study is to describe a Salmonella-monitoring scheme in different food processing plants based on a screening approach by a commercial real-time polymerase chain reaction (PCR) kit and subsequent confirmation of positive molecular results by the reference microbiological method. This scheme was tested on a total of 4,693 samples, 90 of which were positive with the real-time PCR screening; 52 of the positive samples were eventually confirmed by the microbiological method. The real-time PCR kit was tested in comparison to the microbiological method in order to evaluate its performances and drawbacks. The comparison between cycle threshold (Ct) values of real-time PCR and the microbiological results (Wilcoxon rank sum test) showed a statistically significant difference between the Ct values of bacteriological positive and bacteriological negative samples (p value, <0.05). Furthermore, receiver operating characteristic curve analysis was used to identify the Ct value ensuring the lowest level of misclassification between Salmonella-positive and negative samples. The present study confirms that the real-time PCR kit tested could be used as a screening tool, leading to a rapid and sensitive identification of Salmonella and confining bacteriological confirmation to samples previously identified as positive.     

Rheological,mechanical, thermal,and morphological properties of blends poly(butylene adipate-co-terephthalate), thermoplastic starch,and cellulose nanoparticles

The objective of the present study was the preparation and characterization of poly(butylene adipate-co-terephthalate) (PBAT) and thermoplastic starch (TPS) blends reinforced with cellulose nanoparticles (CNCs) by extrusion. The work was conducted in four steps. Initially, the CNCs were prepared from eucalyptus cellulose pulp by acid hydrolysis. The second step was the preparation of the nanocomposite (TPS-CNC), composed of cassava starch, CNC, glycerol, and citric and stearic acids, by double screw extrusion. The third step was the preparation of PBAT/TPS-CNC blends in twin-screw extruders. In the fourth step, the films were produced by flat extrusion. Blends exhibited similar rheological behavior, increasing the CNC concentration in blends increased the viscosity as a function of the shear rate, and altered the behavior of the shear storage (G′) and shear loss (G″) curves as a function of the oscillation frequency (ω). The presence of CNC in blend provided improvements significant in mechanical properties, with 120% increase in Young's modulus, and 46% increase in maximum tensile. Thermal behavior (thermogravimetric analysis and differential scanning calorimetry) was altered with the incorporation of the CNC, showing a single melt peak (T_m) and a slight increase in T_g, indicating good dispersion between the phases of the blends, corroborating with the fracture surface microscopy of films.     

Effect of frozen storage on the phenolic content of vacuum impregnated Granny Smith and Stark Delicious apple <Emphasis Type="Italic">cvv</Emphasis>.

The phenolic fraction modification was evaluated in untreated vacuum impregnated frozen apple slices from two varieties (Granny Smith and Stark Delicious) a few days after freezing and after 12 months of frozen storage. After 12 months the phenolic content of both fresh and vacuum impregnated apple slices varied greatly. The most important change was the increase of many polymeric flavan-3-ols, which may be due to hydrolysis of polymeric procyanidins. Furthermore, there was significant correlation (r ² = 0.52, P < 0.05) between the increase in flavan-3-ols and the decrease in pH. In fact, control Stark slices presented the highest pH, and had no statistically significant increase in flavan-3-ols. Other classes of phenolic compounds were decreased in apple slices following frozen storage.     

Influence of post-casting treatments on sulphonated polyetheretherketone composite membranes

Since the post-casting treatments influence the water entrapped in polymeric matrix and consequently its proton conductivity, an evaluation of annealing at 200 °C and acid treatments was conducted on previously developed composite s-PEEK (1.55 mequiv. g⁻¹) membranes, containing a commercial aminopropyl-functionalised silica. DSC, WAXS, SEM-EDX and laser microscope measurements carried out on membranes swollen at different temperatures highlighted different membrane properties depending on post-casting treatments. It was found that composite membranes have different structural and morphological characteristics than pristine polymer membranes. The silica distribution was modified when different treatments are used. The state of water changed when silica was inserted into the membranes. Actually, contrary to the pristine membranes the presence of freezable water was revealed at temperature lower than 80 °C. The proton conductivity was also affected by the presence and the amount of water trapped into the membranes and was particularly influenced by the post-casting treatments. The silica introduction reduced the swelling effect and improved the robustness of the membranes even if a higher water content in the freezable state was observed. Acid treatment leads to significant improvement in membrane properties, but the present work shows that annealing before acid treatment can affect the membrane morphology more strongly than other treatments resulting in a much better fuel cell performance.     

Hexagonal Voronoi pattern detected in the microstructural design of the echinoid skeleton

Repeated polygonal patterns are pervasive in natural forms and structures. These patterns provide inherent structural stability while optimizing strength-per-weight and minimizing construction costs. In echinoids (sea urchins), a visible regularity can be found in the endoskeleton, consisting of a lightweight and resistant micro-trabecular meshwork (stereom). This foam-like structure follows an intrinsic geometrical pattern that has never been investigated. This study aims to analyse and describe it by focusing on the boss of tubercles—spine attachment sites subject to strong mechanical stresses—in the common sea urchin Paracentrotus lividus. The boss microstructure was identified as a Voronoi construction characterized by 82% concordance to the computed Voronoi models, a prevalence of hexagonal polygons, and a regularly organized seed distribution. This pattern is interpreted as an evolutionary solution for the construction of the echinoid skeleton using a lightweight microstructural design that optimizes the trabecular arrangement, maximizes the structural strength and minimizes the metabolic costs of secreting calcitic stereom. Hence, this identification is particularly valuable to improve the understanding of the mechanical function of the stereom as well as to effectively model and reconstruct similar structures in view of future applications in biomimetic technologies and designs.