SLC6A14, a Pivotal Actor on Cancer Stage: When Function Meets Structure

SLC6A14 (ATB0,+) is a sodium- and chloride-dependent neutral and dibasic amino acid transporter that regulates the distribution of amino acids across cell membranes. The transporter is overexpressed in many human cancers characterized by an increased demand for amino acids; as such, it was recently acknowledged as a novel target for cancer therapy. The knowledge on the molecular mechanism of SLC6A14 transport is still limited, but some elegant studies on related transporters report the involvement of the 12 transmembrane \u3b1-helices in the transport mechanism, and describe structural rearrangements mediated by electrostatic interactions with some pivotal gating residues. In the present work, we constructed a SLC6A14 model in outward-facing conformation via homology modeling and used molecular dynamics simulations to predict amino acid residues critical for substrate recognition and translocation. We docked the proteinogenic amino acids and other known substrates in the SLC6A14 binding site to study both gating regions and the exposed residues involved in transport. Interestingly, some of these residues correspond to those previously identified in other LeuT-fold transporters; however, we could also identify a novel relevant residue with such function. For the first time, by combined approaches of molecular docking and molecular dynamics simulations, we highlight the potential role of these residues in neutral amino acid transport. This novel information unravels new aspects of the human SLC6A14 structure-function relationship and may have important outcomes for cancer treatment through the design of novel inhibitors of SLC6A14-mediated transport

"Delirium Day": A nationwide point prevalence study of delirium in older hospitalized patients using an easy standardized diagnostic tool

Background: To date, delirium prevalence in adult acute hospital populations has been estimated generally from pooled findings of single-center studies and/or among specific patient populations. Furthermore, the number of participants in these studies has not exceeded a few hundred. To overcome these limitations, we have determined, in a multicenter study, the prevalence of delirium over a single day among a large population of patients admitted to acute and rehabilitation hospital wards in Italy. Methods: This is a point prevalence study (called "Delirium Day") including 1867 older patients (aged 65 years or more) across 108 acute and 12 rehabilitation wards in Italian hospitals. Delirium was assessed on the same day in all patients using the 4AT, a validated and briefly administered tool which does not require training. We also collected data regarding motoric subtypes of delirium, functional and nutritional status, dementia, comorbidity, medications, feeding tubes, peripheral venous and urinary catheters, and physical restraints. Results: The mean sample age was 82.0 \ub1 7.5 years (58 % female). Overall, 429 patients (22.9 %) had delirium. Hypoactive was the commonest subtype (132/344 patients, 38.5 %), followed by mixed, hyperactive, and nonmotoric delirium. The prevalence was highest in Neurology (28.5 %) and Geriatrics (24.7 %), lowest in Rehabilitation (14.0 %), and intermediate in Orthopedic (20.6 %) and Internal Medicine wards (21.4 %). In a multivariable logistic regression, age (odds ratio [OR] 1.03, 95 % confidence interval [CI] 1.01-1.05), Activities of Daily Living dependence (OR 1.19, 95 % CI 1.12-1.27), dementia (OR 3.25, 95 % CI 2.41-4.38), malnutrition (OR 2.01, 95 % CI 1.29-3.14), and use of antipsychotics (OR 2.03, 95 % CI 1.45-2.82), feeding tubes (OR 2.51, 95 % CI 1.11-5.66), peripheral venous catheters (OR 1.41, 95 % CI 1.06-1.87), urinary catheters (OR 1.73, 95 % CI 1.30-2.29), and physical restraints (OR 1.84, 95 % CI 1.40-2.40) were associated with delirium. Admission to Neurology wards was also associated with delirium (OR 2.00, 95 % CI 1.29-3.14), while admission to other settings was not. Conclusions: Delirium occurred in more than one out of five patients in acute and rehabilitation hospital wards. Prevalence was highest in Neurology and lowest in Rehabilitation divisions. The "Delirium Day" project might become a useful method to assess delirium across hospital settings and a benchmarking platform for future surveys

Shelf life extension of raspberry : passive and active modified atmosphere inside master bag solutions

Consumers often appreciate small red fruits for their high sensory quality and nutritional value. Furthermore, raspberries are very perishable presenting high respiration rates and a fragile structure; they are easily spoiled by moulds and consequently have a very short shelf life, about 3-5 days. The general purpose of this study was to investigate the ability of passive and active modified atmosphere packaging in extending the shelf life of red raspberries (Rubus idaeus L., cv. Erika). In the case of passive atmosphere, red raspberries were packed inside macro-perforated polyethylene terephthalate (PET) trays inserted in a master bag, made of plastic materials with different permeability and selectivity to oxygen and carbon dioxide. In the case of active packaging, raspberries were packaged into macro-perforated PET trays; 8 of them placed into a cardboard crate, then inserted into an low density polyethylene (LDPE) master bag before sealing, carbon dioxide emitters were introduced. For each trial, the analyses were performed at different times storing the product in a cold dark chamber (51 C, 70 %RH). We performed chemical, mechanical, sensory analyses. Consumer rejection, described by means of the Weibull's equation, was correlated to chemical and physical changes in order to find an acceptability limit useful in defining the product shelf life. The use of active and passive modified atmosphere packaging resulted in a significant increase on the shelf life of raspberries. This will have a great impact on the waste reduction of this product along the distribution chain, increasing its sustainability

Evoluzione della qualità di carne bovina a fette confezionata in atmosfera protettiva in combinazione con assorbitori di ossigeno

Oggetto del presente lavoro \ue8 stata la valutazione della qualit\ue0 di carne bovina a fette (Semitendinosus) durante e in seguito alla conservazione in anossia in un sistema \u201cmaster bag\u201d. Tale sistema di confezionamento prevede l\u2019inserimento di vassoi \u201ccase-ready\u201d e di assorbitori di ossigeno all\u2019interno di un \u201cmaster bag\u201d barriera poi chiuso ermeticamente in atmosfera modificata (30% CO2, 70% N2). La presenza degli assorbitori di ossigeno garantisce il mantenimento dell\u2019anossia per tutta la durata della conservazione nel sacco. Tale condizione \ue8 infatti fondamentale per preservare la qualit\ue0 del prodotto poich\ue9 le basse concentrazioni di ossigeno possono causare l\u2019ossidazione irreversibile dei pigmenti compromettendo quindi l\u2019accettabilit\ue0 del prodotto. Si \ue8 voluto sottoporre alla sperimentazione lo stesso taglio bovino proveniente da due diverse razze (Piemontese e Argentina) con differente storia prima del porzionamento (rispettivamente di frollatura aerobica e sottovuoto, entrambe a 2\ub11\ub0C). Durante tutte le fasi di conservazione (storage life, blooming e display life) sono stati valutati indici microbiologici, colorimetrici e spettrofotometrici (stima delle forme mioglobiniche superficiali), oltre all\u2019acquisizione delle immagini tramite scanner. Entrambe le tipologie di carne sono state conservate in \u201cmaster bag\u201d per almeno 21 giorni, durante i quali non si \ue8 riscontrato un decadimento significativo degli indici qualitativi. Questo risultato pu\uf2 essere anche spiegato dalla funzione sequestrante degli assorbitori: infatti, gi\ue0 a partire dal secondo giorno di conservazione non \ue8 stato pi\uf9 possibile rilevare la presenza di ossigeno all\u2019interno dei sacchi. In seguito all\u2019apertura del \u201cmaster bag\u201d i pigmenti superficiali si riossigenano e conferiscono al prodotto il colore rosso acceso tipico della carne fresca. Grazie alla ripresa della colorazione \ue8 stato possibile esporre i prodotti in un display refrigerato per simularne la vita di scaffale nel punto vendita. L\u2019esposizione nel banco frigo \ue8 stata condotta per 4 giorni durante i quali si \ue8 osservata la graduale ossidazione dei pigmenti superficiali che ha comportato il viraggio del colore verso tonalit\ue0 marroni. La degradazione del prodotto durante la \u201cdisplay life\u201d non \ue8 risultata favorita dal confezionamento in \u201cmaster bag\u201d rispetto a quello tradizionale (tray wrap). \uc8 quindi possibile concludere che il sistema di confezionamento in \u201cmaster bag\u201d garantisce un aumento della vita commerciale di differenti tipologie di carne bovina a fette fino a 21 giorni, senza compromettere l\u2019accettabilit\ue0 del prodotto

Evoluzione della qualità di carne bovina a fette confezionata in atmosfera protettiva in combinazione con assorbitori di ossigeno

Oggetto del presente lavoro \ue8 stata la valutazione della qualit\ue0 di carne bovina a fette (Semitendinosus) durante e in seguito alla conservazione in anossia in un sistema \u201cmaster bag\u201d. Tale sistema di confezionamento prevede l\u2019inserimento di vassoi \u201ccase-ready\u201d e di assorbitori di ossigeno all\u2019interno di un \u201cmaster bag\u201d barriera poi chiuso ermeticamente in atmosfera modificata (30% CO2, 70% N2). La presenza degli assorbitori di ossigeno garantisce il mantenimento dell\u2019anossia per tutta la durata della conservazione nel sacco. Tale condizione \ue8 infatti fondamentale per preservare la qualit\ue0 del prodotto poich\ue9 le basse concentrazioni di ossigeno possono causare l\u2019ossidazione irreversibile dei pigmenti compromettendo quindi l\u2019accettabilit\ue0 del prodotto. Si \ue8 voluto sottoporre alla sperimentazione lo stesso taglio bovino proveniente da due diverse razze (Piemontese e Argentina) con differente storia prima del porzionamento (rispettivamente di frollatura aerobica e sottovuoto, entrambe a 2\ub11\ub0C). Durante tutte le fasi di conservazione (storage life, blooming e display life) sono stati valutati indici microbiologici, colorimetrici e spettrofotometrici (stima delle forme mioglobiniche superficiali), oltre all\u2019acquisizione delle immagini tramite scanner. Entrambe le tipologie di carne sono state conservate in \u201cmaster bag\u201d per almeno 21 giorni, durante i quali non si \ue8 riscontrato un decadimento significativo degli indici qualitativi. Questo risultato pu\uf2 essere anche spiegato dalla funzione sequestrante degli assorbitori: infatti, gi\ue0 a partire dal secondo giorno di conservazione non \ue8 stato pi\uf9 possibile rilevare la presenza di ossigeno all\u2019interno dei sacchi. In seguito all\u2019apertura del \u201cmaster bag\u201d i pigmenti superficiali si riossigenano e conferiscono al prodotto il colore rosso acceso tipico della carne fresca. Grazie alla ripresa della colorazione \ue8 stato possibile esporre i prodotti in un display refrigerato per simularne la vita di scaffale nel punto vendita. L\u2019esposizione nel banco frigo \ue8 stata condotta per 4 giorni durante i quali si \ue8 osservata la graduale ossidazione dei pigmenti superficiali che ha comportato il viraggio del colore verso tonalit\ue0 marroni. La degradazione del prodotto durante la \u201cdisplay life\u201d non \ue8 risultata favorita dal confezionamento in \u201cmaster bag\u201d rispetto a quello tradizionale (tray wrap). \uc8 quindi possibile concludere che il sistema di confezionamento in \u201cmaster bag\u201d garantisce un aumento della vita commerciale di differenti tipologie di carne bovina a fette fino a 21 giorni, senza compromettere l\u2019accettabilit\ue0 del prodotto

Oligosaccharides Related to Tumor-Associated Antigens. Part I. Synthesis of the propyl glycoside of the trisaccharide \u3b1-L-Fucp-(1 \u2192 2)-\u3b2-D-Galp-(1 \u2192 3)-\u3b2-D-GalpNAc, component of a tumor antigen recognized by the antibody MBr1

The synthesis of the trisaccharide alpha-L-Fucp-(1-->2)-beta-D-Galp-(1-->3)-beta-D-GalpNAc-1-OPr (2) is described. The N-acetylgalactosamine 6 was obtained from 4 by an intramolecular displacement of a (trifluoromethyl)sulfonyloxy by a pivaloyloxy group with its concomitant migration from position 3 to position 4 (Scheme 1). The galactosyl donor 9 was obtained from 7 via 8 by regioselective opening of the orthoester function with AcOH/pyridine followed by treatment with CCl3CN and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (Scheme 2). Glycosylation of 6 with 9 in the presence of BF3.OEt2 gave the disaccharide 10. Selective deprotection of 10 at O -C(2') followed by glycosylation with 12 and by standard deprotection afforded the title trisaccharide 2 (Scheme 3). Preliminary biological testing showed that 2 is able to inhibit the binding of the monoclonal antibody MBr1 to the target tumor celles MCF7 in a dose-dependent manner

Acute caffeine administration decreased NOS and Bcl2 expression in rat skeletal muscles

Caffeine (Caf) is largely used to delay fatigue, improving physical activity. However, its role remains elusive, and there are no hemodynamic or immunohistochemical data regarding its effects on skeletal muscle. We studied the hemodynamic and NOS expression of Bax/Bcl2 in skeletal muscle after single Caf administration. Thirty-two male rats were divided into six groups: the first was iv-injected with Caf (16mg/kg), the second with Caf+L-NAME, the third with Caf+L-arg, the fourth with Caf+L-NAME+L-arg, fifth with saline. Mean arterial blood pressure (MAP) was monitored for 30', then the animals were killed. The sixth group was injected with Caf and killed after 2h. The quadriceps were isolated and processed by immunohistochemistry. We found that Caf increased MAP temporarily, while Caf+L-NAME increased it for a longer period. In untreated muscle, all NOS isoforms was expressed with different intensity and localisation, and Bcl2 was strongly expressed among the myofibrils. In Caf and Caf+L-NAME treated animals, NOS expression was lost; Bcl2 expression decreased among myofibrils but increased inside the subsarcolemma. The L-arg administration reversed these Caf and L-NAME effects. Two hours after Caf, NOS expression increased. We concluded that improved physical performance could be related to Caf's ability to interfere with the endogenous muscular synthesis of NO

Acute caffeine administration decreased NOS and Bcl2 expression in rat skeletal muscles.

Caffeine (Caf) is largely used to delay fatigue, improving physical activity. However, its role remains elusive, and there are no hemodynamic or immunohistochemical data regarding its effects on skeletal muscle. We studied the hemodynamic and NOS expression of Bax/Bcl2 in skeletal muscle after single Caf administration. Thirty-two male rats were divided into six groups: the first was iv-injected with Caf (16 mg/kg), the second with Caf + L-NAME, the third with Caf + L-arg, the fourth with Caf + L-NAME+ L-arg, fifth with saline. Mean arterial blood pressure (MAP) was monitored for 30, then the animals were killed. The sixth group was injected with Caf and killed after 2 h. The quadriceps were isolated and processed by immunohistochemistry. We found that Caf increased MAP temporarily, while Caf + L-NAME increased it for a longer period. In untreated muscle, all NOS isoforms was expressed with different intensity and localisation, and Bcl2 was strongly expressed among the myofibrils. In Caf and Caf + L-NAME treated animals, NOS expression was lost; Bcl2 expression decreased among myofibrils but increased inside the subsarcolemma. The L-arg administration reversed these Caf and L-NAME effects. Two hours after Caf, NOS expression increased.We concluded that improved physical performance could be related to Caf’s ability to interfere with the endogenous muscular synthesis of NO