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Percutaneous core biopsy (CB) has been introduced to increase the ability of accurately diagnosing breast malignancies without the need of resorting to surgery. Compared to conventional automated 14 gauge needle core biopsy (NCB), vacuum-assisted needle core biopsy (VANCB) allows obtaining larger specimens and has recognized advantages particularly when the radiological pattern is represented by microcalcifications. Regardless of technical improvements, a small percentage of percutaneous CBs performed to detect breast lesions are still classified, according to European and UK guidelines, in the borderline B3 category, including a group of heterogeneous lesions with uncertain malignant potential. We aimed to assess the prevalence and positive predictive values (PPV) on surgical excision (SE) of B3 category (overall and by sub-categories) in a large series of non-palpable breast lesions assessed through VANCB, also comparison with published data on CB. Overall, 26,165 consecutive stereotactic VANCB were identified in 22 Italian centres: 3107 (11.9%) were classified as B3, of which 1644 (54.2%) proceeded to SE to establish a definitive histological diagnosis of breast pathology. Due to a high proportion of microcalcifications as main radiological pattern, the overall PPV was 21.2% (range 10.6%-27.3% for different B3 subtypes), somewhat lower than the average value (24.5%) from published studies (range 9.9%-35.1%). Our study, to date the largest series of B3 with definitive histological assessment on SE, suggests that B3 lesions should be referred for SE even if VANCB is more accurate than NCB in the diagnostic process of non-palpable, sonographically invisible breast lesions.

Flat epithelial atypia (FEA) may represent the earliest precursor of low-grade breast cancer and often coexists with more advanced atypical proliferative breast lesions such as atypical ductal hyperplasia (ADH) and lobular intraepithelial neoplasia (LIN). The present study aims to investigate the association between morphological parameters of FEA and presence of malignancy at surgical excision (SE) and the clinical significance of the association of FEA with ADH and/or LIN. This study included 589 cases of stereotactic 11-gauge vacuum-assisted needle core biopsy (VANCB), reporting a diagnosis of FEA, ADH or LIN with subsequent SE from 14 pathology departments in Italy. Available slides were reviewed, with 114 (19.4 %) showing a malignant outcome at SE. Among the 190 cases of pure FEA, no statistically significant association emerged between clinical–pathological parameters of FEA and risk of malignancy. Logistic regression analyses showed an increased risk of malignancy according to the extension of ADH among the 275 cases of FEA associated with ADH (p = 0.004) and among the 34 cases of FEA associated with ADH and LIN (p = 0.02). In the whole series, a statistically significant increased malignancy risk emerged according to mammographic R1–R3/R4–R5 categories (OR = 1.56; p = 0.04), extension (OR = 1.24; p = 0.04) and grade (OR = 1.94; p = 0.004) of cytological atypia of FEA. The presence of ADH was associated with an increased malignancy risk (OR = 2.85; p < 0.0001). Our data confirm the frequent association of FEA with ADH and/or LIN. A diagnosis of pure FEA on VANCB carries a 9.5 % risk of concurrent malignancy and thus warrants follow-up excision because none of the clinical–pathological parameters predicts which cases will present carcinoma on SE.

Most of the biological effects displayed by organotin contaminants, among which trisubsituted species are especially toxic, have increasingly been found to exhibit astonishing analogies in different taxa. While similarities can be perceived from prokaryotes to mammals, different modes and extent of biochemical and biological effects were described in different cells, tissues and species. A broad susceptibility range to organotins emerges from literature. Aquatic biota are mainly affected by organotins as environmental water and sediments act as storage site. Endocrine and lipid homeostasis perturbations span from Mollusks, where first gender changes (imposex) referable to environmental organotin contamination was pointed out, to Mammals, where organotins play the role of environmental obesogens. Organotin immunotoxicity, elicited in various invertebrate Phyla, also affects humans. Inhibition of key membrane-bound enzyme complexes such as Na,K-ATPase and FOF1 complexes, thus affecting hydromineral balance, energy production and related effects, are known to occur in a wide variety of organisms. Mitochondria and all membrane functions apparently represent a preferred target of these lipophilic toxicants. Highly conserved action mechanisms could be involved in the observed effects: organotin binding to nuclear receptors, membrane components and intracellular proteins as well as DNA damage may represent widely shared action modes of these compounds. On the other hand the different response and even the refractoriness to these toxicants shown at different biological levels may mirror biochemical and physiological selectivity of signalling pathways, biomembranes and intracellular protein components.

open 6 no The molecular mechanisms which rule the formation and opening of the mitochondrial permeability transition pore (mPTP), the lethal mechanism which permeabilizes mitochondria to water and solutes and drives the cell to death, are still unclear and particularly little investigated in invertebrates. Since Ca2+ increase in mitochondria is accompanied by mPTP opening and the participation of the mitochondrial F1FO-ATPase in the mPTP is increasingly sustained, the substitution of the natural cofactor Mg2+ by Ca2+ in the F1FO-ATPase activation has been involved in the mPTP mechanism. In mussel midgut gland mitochondria the similar kinetic properties of the Mg2+- or Ca2+-dependent F1FO-ATPase activities, namely the same affinity for ATP and bi-site activation kinetics by the ATP substrate, in spite of the higher enzyme activity and coupling efficiency of the Mg2+-dependent F1FO-ATPase, suggest that both enzyme activities are involved in the bioenergetic machinery. Other than being a mitochondrial poison and environmental contaminant, sulfide at low concentrations acts as gaseous mediator and can induce post-translational modifications of proteins. The sulfide donor NaHS, at micromolar concentrations, does not alter the two F1FO-ATPase activities, but desensitizes the mPTP to Ca2+ input. Unexpectedly, NaHS, under the conditions tested, points out a chemical refractoriness of both F1FO-ATPase activities and a failed relationship between the Ca2+-dependent F1FO-ATPase and the mPTP in mussels. The findings suggest that mPTP role and regulation may be different in different taxa and that the F1FO-ATPase insensitivity to NaHS may allow mussels to cope with environmental sulfide. embargoed_20211117 Algieri C.; Nesci S.; Trombetti F.; Fabbri M.; Ventrella V.; Pagliarani A. Algieri C.; Nesci S.; Trombetti F.; Fabbri M.; Ventrella V.; Pagliarani A.

Polar lipids (PoL) represent a new promising dietary approach in the prevention and treatment of many human diseases, due to their potential nutritional value and unique biophysical properties. This study investigates the effects of catching season and oven baking on the fatty acid profiles (FAP) of PoL in four species of blue-back fish widely present in the North Adriatic Sea: anchovy (Engraulis encrasicholus), sardine (Sardina pilchardus), sprat (Sprattus sprattus), and horse mackerel (Trachurus trachurus). PoL levels (427-652 mg/100 g flesh) varied among the four species, with no significant seasonal variations within species. FAP of raw fillets were particularly high in polyunsaturated fatty acid (PUFA), especially docosahexaenoic acid (DHA) and EPA; total PUFA was constant in all species throughout the year, while long-chain n-3 polyunsaturated fatty acid (n-3 PUFA) rose in spring (except in sprat), especially due to the contribution of DHA. The FAP response for PoL to oven baking was species-specific and, among n-3 PUFA, DHA exhibited the greatest heat resistance; the influence of oven baking on FAP was found to be correlated with the catching season, especially for anchovy and sardine, while sprat PoL were not affected by cooking processes. The four species analyzed in this study presented very low n-6/n-3 fatty acid ratios and highly favorable nutritional indices, emphasizing their PoL qualities and promoting their role in increasing human n-3 PUFA intake. The four species can be considered as superior sources of n-3 PUFA and can be employed as supplements in functional food manufacturing and in pharmaceutical and cosmetic industries.

The link between metabolic remodeling and stem cell fate is still unclear. To explore this topic, the metabolic profile of porcine vascular wall mesenchymal stem cells (pVW-MSCs) was investigated. At the first and second cell passages, pVW-MSCs exploit both glycolysis and cellular respiration to synthesize adenosine triphosphate (ATP), but in the subsequent (third to eighth) passages they do not show any mitochondrial ATP turnover. Interestingly, when the first passage pVW-MSCs are exposed to 0.1 or 10 μg/ml lipopolysaccharides (LPSs) for 4 hr, even if ATP synthesis is prevented, the spare respiratory capacity is retained and the glycolytic capacity is unaffected. In contrast, the exposure of pVW-MSCs at the fifth passage to 10 μg/ml LPS stimulates mitochondrial ATP synthesis. Flow cytometry rules out any reactive oxygen species (ROS) involvement in the LPS effects, thus suggesting that the pVW-MSC metabolic pattern is modulated by culture conditions via ROS-independent mechanisms.

Tributyltin (TBT), is a man-made pollutants, known to accumulate along the food chain, acting as an endocrine disruptor in marine organisms, with toxic and adverse effects in many tissues including vascular system. Based on the absence of specific studies of TBT effects on endothelial cells, we aimed to evaluate the toxicity of TBT on primary culture of porcine aortic endothelial cells (pAECs), pig being an excellent model to study human cardiovascular disease. pAECs were exposed for 24h to TBT (100, 250, 500, 750 and 1000nM) showing a dose dependent decrease in cell viability through both apoptosis and necrosis. Moreover the ability of TBT (100 and 500nM) to influence endothelial gene expression was investigated at 1, 7 and 15h of treatment. Gene expression of tight junction molecules, occludin (OCLN) and tight junction protein-1 (ZO-1) was reduced while monocyte adhesion and adhesion molecules ICAM-1 and VCAM-1 (intercellular adhesion molecule-1 and vascular cell adhesion molecule-1) levels increased significantly at 1h. IL-6 and estrogen receptors 1 and 2 (ESR-1 and ESR-2) mRNAs, after a transient decrease, reached the maximum levels after 15h of exposure. Finally, we demonstrated that TBT altered endothelial functionality greatly increasing monocyte adhesion. These findings indicate that TBT deeply alters endothelial profile, disrupting their structure and interfering with their ability to interact with molecules and other cells.

The possibility of increasing n-3 and n-6 long-chain polyunsaturated fatty acids (PUFA) content in microalgal mixtures used to feed Tapes philippinarum larvae was explored by lowering culture temperature from 26 to 14 °C. Although fatty acid composition of different microalgal species has a genetic basis, the algal cultures grown at 14 °C significantly increased the content of long-chain n-3 PUFA in Isocrysis galbana and in Thalassiosira pseudonana, while in Tetraselmis tetrathelo, the PUFA increase only involved shorter chain PUFA, namely 16:4n-3 and 18:4n-3. However, larvae fed on the PUFA enriched microalgal mixture did not show improvements in growth and survival performances with respect to the control group fed the microalgal mixture grown at 26 °C. From a biochemical perspective, two key aspects emerged from the results: (i) clam larvae have adequate biotransformation and selection skills to adjust fatty acid profile to their requirements as they can even modulate the incorporation of essential long-chain PUFA as 20:5n-3 and 22:6n-3 when the dietary supply exceeds the physiological requirements; (ii) bivalve can biosynthesize non-methylene-interrupted dienoic (NMID) fatty acids as confirmed by the constancy of relative proportion with larvae growth in spite of the NMID fatty acid absence in the diet.

Increasing evidence highlights the role of the ATP synthase/hydrolase, also known as F1FO-complex, as key molecular and enzymatic switch between cell life and death, thus increasing the enzyme attractiveness as drug target in pharmacology. Being inhibition of ATP production usually linked to antiproliferative properties, drugs targeting the enzyme complex have been mainly considered to fight pathogen parasites and cancer. In recent years, a number of natural macrolides, produced by bacterial fermentation and structurally related to the classical enzyme inhibitor oligomycin, have been shown to bind to the membrane-embedded FO sector and to inhibit the enzyme complex by an oligomycin-like mechanism, namely by interacting with the c-ring. Other than natural macrolide antibiotics, which display variegated inhibition power on different F1FO-complexes, synthetic compounds from the diarylquinoline and organotin families also target the c-ring and strongly inhibit the enzyme. Bioinformatic insights address drug design to target FO subunits. Additionally, the possible modulation of the drug inhibition power, by amino acid substitutions or post-translational modifications of c-subunits, adds further interest to the target. The present survey on compounds targeting the c-ring and bi-directionally blocking the transmembrane proton flux which drives ATP synthesis/hydrolysis, discloses new therapeutic options to fight cancer and infections sustained by therapeutically recalcitrant microorganisms. Additionally, c-ring targeting compounds may constitute new tools to eradicate undesired biofilms and to address at the molecular level the therapy of mammalian diseases linked to mitochondrial dysfunctions. In summary, studies on the only partially known molecular interactions within the c-ring of the F1FO-complex may renew hope to counteract mammalian diseases.

The diarrhetic poisoning by bivalve molluscs, diarrhetic shellfish poisoning, is due to consumption of mussels containing biotoxins produced by some Dinoflagellate species. Toxic effects of yessotoxin (YTX) include morphological alterations of mitochondria from heart and liver but the biochemical basis for these alterations is completely unknown.This paper demonstrates that YTX is a very powerful compound that opens the permeability transition pore (PTP) of the inner mitochondrial membrane of rat liver mitochondria at nanomolar concentrations. The effect requires the presence of a permissive level of calcium, by itself incapable of opening the pore. The direct effect of YTX on PTP is further confirmed by the inhibition exerted by cyclosporin A (CsA) that is known as a powerful inhibitor of PTP opening. Moreover, YTX induces membrane depolarization as shown by the quenching of tetramethylrhodamine methyl ester (TMRM), also prevented by the addition of CsA. YTX caused PTP opening in Morris Hepatoma 1C1 cells, as shown by the occurrence of CsA-sensitive depolarization within minutes of the addition of submicromolar concentrations of the toxin. These results provide a biochemical basis for the mitochondrial alterations observed in the course of intoxication with YTX, offering the first clue into the pathogenesis of diseases caused by YTX, and providing a novel tool to study the PTP in situ.