Discovery of a septin-4 covalent binder with antimetastatic activity in a mouse model of melanoma.

Cancer spreading through metastatic processes is one of the major causes of tumour-related mortality. Metastasis is a complex phenomenon which involves multiple pathways ranging from cell metabolic alterations to changes in the biophysical phenotype of cells and tissues. In the search for new effective anti-metastatic agents, we modulated the chemical structure of the lead compound AA6, in order to find the structural determinants of activity, and to identify the cellular target responsible of the downstream anti-metastatic effects observed. New compounds synthesized were able to inhibit in vitro B16-F10 melanoma cell invasiveness, and one selected compound, CM365, showed in vivo anti-metastatic effects in a lung metastasis mouse model of melanoma. Septin-4 was identified as the most likely molecular target responsible for these effects. This study showed that CM365 is a promising molecule for metastasis prevention, remarkably effective alone or co-administered with drugs normally used in cancer therapy, such as paclitaxel.

New lipophilic organic nitrates: candidates for chronic skin disease therapy.

Organic nitrates are widely used, but their chronic efficacy is blunted due to the development of tolerance. The properties of new tolerance free organic nitrates were studied. Their lipophilicity profile and passive diffusion across polydimethylsiloxane membrane and pig ear-skin, and their efficacy in tissue regeneration using HaCaT keratinocytes were evaluated. The permeation results show that these nitrates have a suitable profile for NO topical administration on the skin. Furthermore, the derivatives with higher NO release exerted a pro-healing effect on HaCaT cells. This new class of organic nitrates might be a promising strategy for the chronic treatment of skin pathologies.

Bioelectrical impedance analysis versus reference methods in the assessment of body composition in athletes.

The present systematic review aimed to compare the accuracy of Bioelectrical Impedance Analysis (BIA) and Bioelectrical Impedance Vector Analysis (BIVA) vs. reference methods for the assessment of body composition in athletes. Studies were identified based on a systematic search of internationally electronic databases (PubMed and Scopus) and hand searching of the reference lists of the included studies. In total, 42 studies published between 1988 and 2021 were included. The methodological quality was assessed using the Quality Assessment Tool for Observational Cohort and Cross-sectional Studies as recommended by the National Institute of Health. Twenty-three studies had an overall good rating in terms of quality, while 13 were rated as fair and 6 as poor, resulting in a low to moderate risk of bias. Fat mass was inconsistently determined using BIA vs. the reference methods, regardless of the BIA-technology. When using the foot to hand technology with predictive equations for athletes, a good agreement between BIA and the reference methods was observed for fat-free mass, total body, intra and extra cellular water. However, an underestimation in fat-free mass and body fluids was found when using generalized predictive equations. Classic and Specific BIVA represented a valid approach for assessing body fluids (Classic BIVA) and percentage of fat mass (Specific BIVA). The present systematic review suggests that BIA and BIVA can be used for assessing body composition in athletes, provided that foot-to-hand technology, predictive equations, and BIVA references for athletes are used.

Fat-Free Mass Using Bioelectrical Impedance Analysis as an Alternative to Dual-Energy X-Ray Absorptiometry in Calculating Energy Availability in Female Adolescent Athletes.

Energy availability (EA) is calculated by subtracting exercise energy expenditure from energy intake, adjusted for fat-free mass (FFM) obtained using accurate methods, such as dual-energy X-ray absorptiometry (DXA). Unlike DXA, the bioelectrical impedance analysis (BIA) is low in cost, simple and easy to carry out. This study aimed to test the concordance between the calculation of EA using FFM values from four BIA predictive equations and FFM obtained using DXA in female adolescent athletes (n = 94), recruited via social media. Paired Student's t test, Wilcoxon test, Lin's concordance correlation coefficient, root mean square error, limits of agreement, and mean absolute percentage error were used to evaluate agreement between the FFM values obtained by the four SF-BIA predictive equations and DXA. Regression linear analysis was used to determine the relation between FFM values obtained using DXA and the BIA predictive equations. Standardized residuals of the FFM and EA were calculated considering DXA values as reference. The most appropriate model for the FFM (limits of agreement = 4.0/-2.6 kg, root mean square error = 1.9 kg, mean absolute percentage error = 4.34%, Lin's concordance correlation coefficient = .926) and EA (limits of agreement = 2.51/4.4 kcal·kg FFM-1·day-1, root mean square error = 1.8 kcal·kg FFM-1·day-1, mean absolute percentage error 4.24%, Lin's concordance correlation coefficient = .992) was the equation with sexual maturity as a variable, while the equation with the greatest age variability was the one with the lowest agreement. FFM-BIA predictive equations can be used to calculate EA of female adolescent athletes. However, the equation should be chosen considering sex, age, and maturation status. In the case of athletes, researchers should use equations developed for this group.

Investigation into the Use of Encorafenib to Develop Potential PROTACs Directed against BRAFV600E Protein.

BRAF is a serine/threonine kinase frequently mutated in human cancers. BRAFV600E mutated protein is targeted through the use of kinase inhibitors which are approved for the treatment of melanoma; however, their long-term efficacy is hampered by resistance mechanisms. The PROTAC-induced degradation of BRAFV600E has been proposed as an alternative strategy to avoid the onset of resistance. In this study, we designed a series of compounds where the BRAF kinase inhibitor encorafenib was conjugated to pomalidomide through different linkers. The synthesized compounds maintained their ability to inhibit the kinase activity of mutated BRAF with IC50 values in the 40-88 nM range. Selected compounds inhibited BRAFV600E signaling and cellular proliferation of A375 and Colo205 tumor cell lines. Compounds 10 and 11, the most active of the series, were not able to induce degradation of mutated BRAF. Docking and molecular dynamic studies, conducted in comparison with the efficient BRAF degrader P5B, suggest that a different orientation of the linker bearing the pomalidomide substructure, together with a decreased mobility of the solvent-exposed part of the conjugates, could explain this behavior.

Tuning NO release of organelle-targeted furoxan derivatives and their cytotoxicity against lung cancer cells.

We herein report a study on a set of hybrid compounds in which 3-R-substituted furoxan moieties (R = CH3, CONH2, CN, SO2C6H5), endowed with varying NO-releasing capacities, are joined to a mitochondrial probe, rhodamine B. Each product has been investigated for its ability to release NO both in physiological solution, in the presence of cysteine, and in A549 lung adenocarcinoma cancer cells. The cytotoxicity of all the products against the aforementioned cancer cells has been assessed, including the structurally related compounds with no mitochondrial targeting, which were taken as a reference. In the case of the models bearing the -CH3 and -CONH2 groups at the 3-position on the furoxan, only the targeted models showed a significant cytotoxic activity, and only at the highest concentrations, in accordance with their weak NO-releasing properties. On the contrary, the presence of the strong electron-withdrawing groups, as -CN and -SO2C6H5, at the 3-position gave rise to anticancer agents, likely because of the high NO-releasing and of their capability of inhibiting cellular proteins by covalent binding. In detail, the rhodamine hybrid containing the 3-SO2C6H5 substituted furoxan moiety emerged as the most interesting product as it showed high cytotoxicity over the entire concentration range tested. This substructure was also linked to a phenothiazine scaffold that is able to accumulate in lysosomes. Nevertheless, mitochondrial targeting for these NO-donor furoxan substructures was found to be the most efficient.

Glargine insulin loaded lipid nanoparticles: Oral delivery of liquid and solid oral dosage forms.

BACKGROUND AND AIMS: The oral administration of insulin has so far been precluded by gastro-intestinal enzyme degradation and poor intestinal absorption. Preliminary evidence for peptide uptake by the gut has recently been obtained, by our research group, following the administration of nanostructured lipid-carrier suspensions loaded with glargine insulin in healthy animal models. METHODS AND RESULTS: In this experimental study, glargine insulin-loaded nanostructured lipid carriers have been converted into solid oral dosage forms (tablets, capsules), that are more suitable for administration in humans and have prolonged shelf-life. The liquid and solid oral dosage forms were tested for glargine insulin uptake and glucose responsivity in healthy and streptozotocin-induced diabetic rats (6 animals in each group). A suitable composition gave redispersible solid oral dosage forms from glargine insulin-loaded carriers, using both spray-drying and freeze-drying. It was observed that the liquid and solid formulations had relevant hypoglycaemic effects in healthy rats, while only capsules were efficacious in diabetic rats; probably because of gut alterations in these animal models. Detected glargine insulinaemia was consistent with a glycaemic profile. CONCLUSION: The formulations under study showed their potential as oral glucose-lowering agents, particularly when used as capsules. However, further study is needed to produce a useful orally-active insulin preparation.

Chemical Modulation of the 1-(Piperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazole-2-one Scaffold as a Novel NLRP3 Inhibitor.

In the search for new chemical scaffolds able to afford NLRP3 inflammasome inhibitors, we used a pharmacophore-hybridization strategy by combining the structure of the acrylic acid derivative INF39 with the 1-(piperidin-4-yl)1,3-dihydro-2H-benzo[d]imidazole-2-one substructure present in HS203873, a recently identified NLRP3 binder. A series of differently modulated benzo[d]imidazole-2-one derivatives were designed and synthesised. The obtained compounds were screened in vitro to test their ability to inhibit NLRP3-dependent pyroptosis and IL-1ß release in PMA-differentiated THP-1 cells stimulated with LPS/ATP. The selected compounds were evaluated for their ability to reduce the ATPase activity of human recombinant NLRP3 using a newly developed assay. From this screening, compounds 9, 13 and 18, able to concentration-dependently inhibit IL-1ß release in LPS/ATP-stimulated human macrophages, emerged as the most promising NLRP3 inhibitors of the series. Computational simulations were applied for building the first complete model of the NLRP3 inactive state and for identifying possible binding sites available to the tested compounds. The analyses led us to suggest a mechanism of protein-ligand binding that might explain the activity of the compounds.

The relationship among the age at menarche, anthropometric characteristics, and socio-economic factors in Bengali girls from Kolkata, India.

OBJECTIVES: The aim of the study was to measure the causal effect of selected socio-economic factors and anthropometrical characteristics on the menarche occurrence. METHODS: The sample consisted of 2195 Bengali girls (aged 7-21) from middle-class families, from Kolkata city, India. The age at menarche was recorded from the retrospective data and status quo method. The causal effect of anthropometric and socio-economic variables on menarche occurrence was estimated by the nonparametrical analysis of survival probability (survival random forest). RESULTS: In the examined cohort menarche occurred, on average, at 11.8 years of age. The probability of menarche occurrence amplified with the increasing values of factors such as body mass index, height-for-age z-scores, number of family members, household rooms, and toilets, but decreased when expenditures increased. The relation maintained a similar pattern of causal effect with girls' age. CONCLUSIONS: A complex pattern of relationship among sexual development, physique, and socio-economic characteristics was defined. The tendency toward early menarche, along with the observed causal relationships indicate that the analyzed sample is nearing the characteristics and standards of living noted in other middle and even high-income countries in the world.

Development of covalent NLRP3 inflammasome inhibitors: Chemistry and biological activity.

The NOD-like receptor family, pyrin domain-containing 3 (NLRP3) inflammasome is the best recognized and most widely implicated regulator of caspase-1 activation. It is a key regulator of innate immune response and is involved in many pathophysiological processes. Recent evidences for its inappropriate activation in autoinflammatory, autoimmune, as well as in neurodegenerative diseases attract a growing interest toward the development of small molecules NLRP3 inhibitors. Based on the knowledge of biochemical and structural aspects of NLRP3 activation, one successful strategy in the identification of NLRP3 inhibitors relies on the development of covalent irreversible inhibitors. Covalent inhibitors are reactive electrophilic molecules able to alkylate nucleophiles in the target protein. These inhibitors could ensure good efficacy and prolonged duration of action both in vitro and in vivo. In spite of these advantages, effects on other signalling pathways, prone to alkylation, may occur. In this review, we will illustrate the chemistry and the biological action of the most studied covalent NLRP3 inhibitors developed so far. A description of what we know about their mechanism of action will address the reader toward a critical understanding of NLRP3 inhibition by electrophilic compounds.

Nutritional status of Ugandan school-children: The effect of age imprecision.

OBJECTIVES: To analyze the nutritional status of Ugandan school-children in a cross-sectional and longitudinal perspective, considering the effect of age imprecision. MATERIALS AND METHODS: Anthropometric measurements of 831 school-children (381 males and 450 females) were analyzed. A subsample of 246 children was measured in July 2014 and 2015. Stunting (based on height-for-age Z-scores), underweight (weight-for-age), and thinness (body mass index-for-age) prevalence were calculated. Three different ages were used: declared (from schools registers), attributed (based on multiple information sources), and bootstrap (from 10,000 replicates). Significant differences among malnutrition prevalence calculated with different ages and in different groups were assessed by means of bootstrap analysis. Longitudinal analysis was conducted using a paired t test. RESULTS: The mean prevalence of malnutrition calculated with declared, attributed, or bootstrap ages were very similar: stunting (11.9-12.7); underweight (5.4-5.9); thinness (3.3-3.7); and obesity (0.7). Undernutrition was more prevalent among older children, while obesity was mostly associated with young age. Obesity was equally distributed among sexes, while undernutrition was more prevalent among females of up to 10 years of age and males above 10 years. The longitudinal analysis indicated a reduction in underweight and thinness, and an increase in stunting, especially among older children. DISCUSSION: Age imprecision did not significantly affect malnutrition estimates. Despite the decline in the prevalence of thinness and underweight observed over a 1-year period, undernutrition persists, with an observed rise in stunting. On the other hand, obesity is starting to appear. Public health efforts are required to eliminate stunting and address the emerging burden of obesity.

Aceclofenac-Galactose Conjugate: Design, Synthesis, Characterization, and Pharmacological and Toxicological Evaluations.

Aceclofenac is a popular analgesic, antipyretic, and nonsteroidal anti-inflammatory drug (NSAID) used for prolonged treatment (at least three months) in musculoskeletal disorders. It is characterized by several limitations such as poor water solubility and low oral bioavailability. The main side-effect of aceclofenac, as well as all NSAIDs, is the gastrotoxicity; among other adverse effects, there is the risk of bleeding since aceclofenac reversibly inhibits platelet aggregation. With the aim to reduce these drawbacks, we have designed, synthesized, and characterized, both in vitro and in vivo, an orally administrable pro-drug of aceclofenac (ACEgal). ACEgal was obtained by conjugating carboxyl group with the 6-OH group of d-galactose; its structure was confirmed by X-ray powder diffractometry. The pro-drug was shown to be stable at 37 °C in simulated gastric fluid (SGF-without pepsin, pH = 1.2) and moderately stable in phosphate buffered saline (PBS, pH = 7.4). However, it hydrolyzed in human serum with a half-life ( t1/2) of 36 min, producing aceclofenac. Furthermore, if compared to its parent drug, ACEgal was four-times more soluble in SGF. To predict human intestinal absorption, cell permeability in a Caco-2 model of aceclofenac and ACEgal was determined. Anti-inflammatory, analgesic, and ulcerogenic activities have been investigated in vivo. In addition, oxidative stress parameters (thiobarbituric acid reactive substances, TBARS, and glutathione, GSH) and platelet antiaggregatory activity both of parent drug and pro-drug were evaluated. Results clearly showed that the conjugation of aceclofenac to a galactose molecule improves physicochemical, toxicological (at gastric and blood level), and pharmacological profile of aceclofenac itself without changing intestinal permeability and antiplatelet activity (in spite the new sugar moiety).

Cross-sectional and longitudinal analysis of nutritional status of school-children from Bumbire Island (United Republic of Tanzania).

OBJECTIVE: To analyze the nutritional status of schoolchildren from Bumbire Island (Tanzania) from cross-sectional and longitudinal perspectives. METHODS: During 2014 and 2015, we collected anthropometric measurements in a sample of 437 schoolchildren (226 males, 211 females; 5-16 years). A sub-sample of 126 children were measured in both surveys. Socio-demographic data have been taken and dietary habits investigated. The accuracy of age data was checked. Malnutrition prevalence was calculated according to the WHO references and the z-score criteria. RESULTS: The prevalence of undernutrition was high (stunting: 30.7%; underweight: 12.9%; thinness: 4.5%), while overweight was rare (2.4%). The prevalence of stunting was higher in males and in older children. The one-year longitudinal analysis indicated that stunting prevalence increased. CONCLUSIONS: Undernutrition is affecting Bumbire Island children, likely due to micronutrient deficiencies. The effects of linear growth deficit continue to accumulate throughout childhood and adolescent years.

A Nonmetal-Containing Nitric Oxide Donor Activated with Single-Photon Green Light.

Using a facile synthetic route, an organic NO release agent based on a BODIPY light-harvesting antenna was devised. This compound is stable in the dark and delivers NO under photoexcitation with biologically favorable green light. Temporally regulated vasodilation capability is demonstrated on rat aorta by green-light-induced NO release.

Phylogenetic and ecological factors impact the gut microbiota of two Neotropical primate species.

Recent studies suggest that variation in diet across time and space results in changes in the mammalian gut microbiota. This variation may ultimately impact host ecology by altering nutritional status and health. Wild animal populations provide an excellent opportunity for understanding these interactions. However, compared to clinical studies, microbial research targeting wild animals is currently limited, and many published studies focus only on a single population of a single host species. In this study we utilize fecal samples from two species of howler monkey (Alouatta pigra and A. palliata) collected at four sites to investigate factors influencing the gut microbiota at three scales: taxonomic (host species), ecosystemic (forest type), and local (habitat disturbance/season). The results demonstrate that the effect of host species on the gut microbiota is stronger than the effect of host forest type, which is stronger than the effect of habitat disturbance or seasonality. Nevertheless, within host species, gut microbiota composition differs in response to forest type, habitat disturbance, and season. Variations in the effect size of these factors are associated both with host species and environment. This information may be beneficial for understanding ecological and evolutionary questions associated with Mesoamerican howler monkeys, as well as determining conservation challenges facing each species. These mechanisms may also provide insight into the ecology of other species of howler monkeys, non-human primates, and mammals.

Birth registration and child undernutrition in sub-Saharan Africa.

OBJECTIVE: In many countries of the world millions of people are not registered at birth. However, in order to assess children's nutritional status it is necessary to have an exact knowledge of their age. In the present paper we discuss the effects of insufficient or imprecise age data on estimates of undernutrition prevalence. DESIGN: Birth registration rates and levels of stunting, underweight and wasting were retrieved from Multiple Indicator Cluster Surveys and Demographic and Health Surveys of thirty-seven sub-Saharan African countries, considering the subdivision in wealth quintiles. The composition of the cross-sectional sample used for nutritional evaluation was analysed using a permutation test. Logistic regression was applied to analyse the relationship between birth registration and undernutrition. The 95 % probability intervals and Student's t test were used to evaluate the effect of age bias and error. RESULTS: Heterogeneous sampling designs were detected among countries, with different percentages of children selected for anthropometry. Further, registered children were slightly more represented within samples used for nutritional analysis than in the total sample. A negative relationship between birth registration and undernutrition was recognized, with registered children showing a better nutritional status than unregistered ones, even within each wealth quintile. The over- or underestimation of undernutrition in the case of systematic over- or underestimation of age, respectively, the latter being more probable, was quantified up to 28 %. Age imprecision was shown to slightly overestimate undernutrition. CONCLUSIONS: Selection bias towards registered children and underestimation of children's age can lead to an underestimation of the prevalence of undernutrition.

NO-donor thiacarbocyanines as multifunctional agents for Alzheimer's disease.

Some symmetrical and unsymmetrical thiacarbocyanines bearing NO-donor nitrooxy and furoxan moieties were synthesized and studied as candidate anti-Alzheimer's drugs. All products activated soluble guanylate cyclase (sGC) in a dose-dependent manner, depending on the presence in their structures of NO-donor groups. None displayed toxicity when tested at concentrations below 10 µM on human brain microvascular endothelial cells (hCMEC/D3). Some products were capable of inhibiting amyloid ß-protein (Aß) aggregation, with a potency in the low µM concentration range, and of inhibiting aggregation of human recombinant tau protein in amyloid fibrils when incubated with the protein at 1 µM concentration. Nitrooxy derivative 21 and furoxan derivative 22 were selected to investigate synaptic plasticity. Both products, tested at 2 µM concentration, counteracted the inhibition of long-term potentiation (LTP) induced by Aß42 in hippocampal brain slices.

New specific bioelectrical impedance vector reference values for assessing body composition in the Italian-Spanish young adult population.

OBJECTIVE: Specific bioelectrical impedance vector analysis (spBIVA) is a recently proposed technique for the analysis of body composition. The aim of this study was to apply spBIVA to a sample of Italian and Spanish young adults and to define the new bioelectrical references for this Western Mediterranean population. METHODS: A sample of 440 individuals (220 from Italy, 220 from Spain; 213 men, 227 women) aged 18-30 years was considered. Anthropometric (height, weight, relaxed upper arm, waist, and calf girths) and bioelectrical (resistance, reactance; 50 kHz, 800 µA) measurements were taken. In order to verify the need for new references, specific bioelectrical values were compared to the reference values for U.S. adults and Italian elderly by tolerance ellipses and Student's t test. RESULTS: The mean specific bioelectrical values (resistivity, Rsp, and reactivity, Xcsp, Ohm·cm) were: Rsp (332.7 ± 41.7 Ω·cm), Xcsp (44.4 ± 6.8 Ω·cm), Zsp (335.6 ± 41.9 Ω·cm) and phase (7.6 ± 0.8°) in men; Rsp (388.6 ± 60 Ω·cm), Xcsp (43.7 ± 7.5 Ω·cm), Zsp (391.0 ± 60.3 Ω·cm) and phase (6.4 ± 0.7°) in women. Italo-Spanish bioelectrical vectors were mainly distributed (>90%) in the lower part of the tolerance ellipses for U.S. young adults, due to a shorter impedance (P < 0.001), indicative of a lower percent fat mass. Compared to Italian elders, they were mainly located in the left side (>90%), due to a higher phase (P < 0.001), indicative of higher body cell mass. CONCLUSIONS: These population and age-related differences indicate the need for new specific tolerance ellipses that can be used as references for assessing body composition in young adults from Western Mediterranean populations.

A new, effective and low-cost three-dimensional approach for the estimation of upper-limb volume.

The aim of this research was to validate a new procedure (SkanLab) for the three-dimensional estimation of total arm volume. SkanLab is based on a single structured-light Kinect sensor (Microsoft, Redmond, WA, USA) and on Skanect (Occipital, San Francisco, CA, USA) and MeshLab (Visual Computing Lab, Pisa, Italy) software. The volume of twelve plastic cylinders was measured using geometry, as the reference, water displacement and SkanLab techniques (two raters and repetitions). The right total arm volume of thirty adults was measured by water displacement (reference) and SkanLab (two raters and repetitions). The bias and limits of agreement (LOA) between techniques were determined using the Bland-Altman method. Intra- and inter-rater reliability was assessed using the intraclass correlation coefficient (ICC) and the standard error of measurement. The bias of SkanLab in measuring the cylinders volume was -21.9 mL (-5.7%) (LOA: -62.0 to 18.2 mL; -18.1% to 6.7%) and in measuring the volume of arms' was -9.9 mL (-0.6%) (LOA: -49.6 to 29.8 mL; -2.6% to 1.4%). SkanLab's intra- and inter-rater reliabilities were very high (ICC >0.99). In conclusion, SkanLab is a fast, safe and low-cost method for assessing total arm volume, with high levels of accuracy and reliability. SkanLab represents a promising tool in clinical applications.