Immobilization of Monolayer Protected Lipophilic Gold Nanorods on a Glass Surface

We present a novel process of immobilization of gold nanorods (GNRs) on a glass surface. Wedemonstrate that by exploiting monolayer protection of the GNRs, their unusual opticalproperties can be completely preserved. UV–visible spectroscopy and atomic forcemicroscopy analysis are used to reveal the optical and morphological properties of monolayerprotected immobilized lipophilic GNRs, and molecular dynamics simulations are used toelucidate their surface molecule arrangements

Training Effects on ROS Production Determined by Electron Paramagnetic Resonance in Master Swimmers

Acute exercise induces an increase in Reactive Oxygen Species (ROS) production dependent on exercise intensity with highest ROS amount generated by strenuous exercise. However, chronic repetition of exercise, that is, exercise training, may reduce exercise-induced oxidative stress. Aim of this study was to evaluate the effects of 6-weeks high-intensity discontinuous training (HIDT), characterized by repeated variations of intensity and changes of redox potential, on ROS production and antioxidant capacity in sixteen master swimmers. Time course changes of ROS generation were assessed by Electron Paramagnetic Resonance in capillary blood by a microinvasive approach. An incremental arm-ergometer exercise (IE) until exhaustion was carried out at both before (PRE) and after (POST) training (Trg) period. A significant (P<0.01) increase of ROS production from REST to the END of IE in PRE Trg (2.82±0.66 versus 3.28±0.66 µmol·min−1) was observed. HIDT increased peak oxygen consumption (36.1±4.3 versus 40.6±5.7 mL·kg−1·min−1 PRE and POST Trg, resp.) and the antioxidant capacity (+13%) while it significantly decreased the ROS production both at REST (−20%) and after IE (−25%). The observed link between ROS production, adaptive antioxidant defense mechanisms, and peak oxygen consumption provides new insight into the correlation between ROS response pathways and muscle metabolic function

The Role of Pathological Method and Clearance Definition for the Evaluation of Margin Status after Pancreatoduodenectomy for Periampullary Cancer. Results of a Multicenter Prospective Randomized Trial

Simple SummaryThere is no clear evidence on the most effective method of pathological analysis and clearance definition (0 vs. 1 mm) to define R1 resection after pancreatoduodenectomy (PD). However, several studies showed that the R1 resection is a poor prognostic factor in patients that have undergone PDs for periampullary cancers. In this randomized clinical trial, specimens were randomized with two pathological methods, the Leeds Pathology Protocol (LEEPP) or the conventional method adopted before the study. The 1 mm clearance is the most effective factor in determining R1 rate after PD but only when adopting the LEEP, the R1 resection represents a significant prognostic factor.Background: There is extreme heterogeneity in the available literature on the determination of R1 resection rate after pancreatoduodenectomy (PD); consequently, its prognostic role is still debated. The aims of this multicenter randomized study were to evaluate the effect of sampling and clearance definition in determining R1 rate after PD for periampullary cancer and to assess the prognostic role of R1 resection. Methods: PD specimens were randomized to Leeds Pathology Protocol (LEEPP) (group A) or the conventional method adopted before the study (group B). R1 rate was determined by adopting 0- and 1-mm clearance; the association between R1, local recurrence (LR) and overall survival (OS) was also evaluated. Results. One-hundred-sixty-eight PD specimens were included. With 0 mm clearance, R1 rate was 26.2% and 20.2% for groups A and B, respectively; with 1 mm, R1 rate was 60.7% and 57.1%, respectively (p &gt; 0.05). Only in group A was R1 found to be a significant prognostic factor: at 0 mm, median OS was 36 and 20 months for R0 and R1, respectively, while at 1 mm, median OS was not reached and 30 months. At multivariate analysis, R1 resection was found to be a significant prognostic factor independent of clearance definition only in the case of the adoption of LEEPP. Conclusions. The 1 mm clearance is the most effective factor in determining the R1 rate after PD. However, the pathological method is crucial to accurately evaluate its prognostic role: only R1 resections obtained with the adoption of LEEPP seem to significantly affect prognosis

Assessment of a Standardized ROS Production Profile in Humans by Electron Paramagnetic Resonance

Despite the growing interest in the role of reactive oxygen species (ROS) in health and disease, reliable quantitative noninvasive methods for the assessment of oxidative stress in humans are still lacking. EPR technique, coupled to a specific spin probe (CMH: 1-hydroxy-3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine) is here presented as the method of choice to gain a direct measurement of ROS in biological fluids and tissues. The study aimed at demonstrating that, differently from currently available “a posteriori” assays of ROS-induced damage by means of biomolecules (e.g., proteins and lipids) spin-trapping EPR provides direct evidence of the “instantaneous” presence of radical species in the sample and, as signal areas are proportional to the number of excited electron spins, lead to absolute concentration levels. Using a recently developed bench top continuous wave system (e-scan EPR scanner, Bruker) dealing with very low ROS concentration levels in small (50 μL) samples, we successfully monitored rapid ROS production changes in peripheral blood of athletes after controlled exercise and sedentary subjects after antioxidant supplementation. The correlation between EPR results and data obtained by various enzymatic assays (e.g., protein carbonyls and thiobarbituric acid reactive substances) was determined too. Synthetically, our method allows reliable, quick, noninvasive quantitative determination of ROS in human peripheral blood

BASI MOLECOLARI E CELLULARI DELLA VITA

Prefazione La biologia è … un campo dalle possibilità illimitate, dal quale ci dobbiamo attendere le più sorprendenti delucidazioni; non possiamo quindi indovinare quali risposte essa potrà dare … ai problemi che le abbiamo posto. Forse queste risposte saranno tali da far crollare tutto l’artificioso edificio delle nostre ipotesi. (Sigmund Freud) La Biologia ha compiuto passi da gigante nel tempo intercorso tra la predizione di Freud e i nostri giorni. Nonostante ciò, il suo insegnamento a livello universitario, relativamente a molti corsi di laurea nei quali non costituisce materia fondante, vive un momento di grande incertezza. Infatti, le riforme degli ordinamenti con la suddivisione dei corsi di laurea in triennali e specialistici prima e magistrali poi, e l’introduzione dei corsi di laurea professionalizzanti hanno costretto molte facoltà a rivedere profondamente e radicalmente le loro offerte formative. Nella maggior parte dei casi, è proposto un numero eccessivo di insegnamenti da impartire in poche ore e con contenuti disciplinari ancora non ben definiti, con l’idea che la laurea triennale debba avere la capacità di preparare gli studenti ad affacciarsi al mondo del lavoro. L’organizzazione degli insegnamenti in moduli integrati, cioè insiemi di attività caratterizzati da una certa coerenza, pur essendo guidata dall’obiettivo condivisibile di impartire una preparazione di ampio respiro, ha rappresentato una fonte di problemi nell’identificazione dei programmi didattici relativi a ogni disciplina. Ciò richiede ancora, in molti casi, un’azione sensibilizzata alla selezione degli argomenti da trattare in ogni modulo e all’integrazione con le discipline affini. In questo contesto e in quanto impegnati in attività didattica in questi corsi, abbiamo sentito la necessità di tarare accuratamente l’insegnamento della Biologia, che copre un insieme enorme di argomenti, per adattarlo alle nuove esigenze. “Basi molecolari e cellulari della vita” è frutto di uno sforzo critico che ha permesso di rivedere i contenuti fondamentali della Biologia, indirizzandoli in modo specifico alla formazione di base delle figure destinate ad operare in molti casi in un ambiente clinico e al diretto contatto con i pazienti. Nella sua struttura, il volume è adatto a coprire la preparazione per le professioni sanitarie, quali ad es. quelle infermieristiche e riabilitative, le professioni tecnico-sanitarie, le professioni sanitarie tecniche della prevenzione, ma è stato formulato anche con l’obiettivo di soddisfare le esigenze degli studenti dei corsi di laurea triennale in scienze motorie. Gli argomenti trattati guidano gli studenti, indipendentemente dalla preparazione acquisita con gli studi secondari, alla comprensione degli indispensabili argomenti di base della struttura e delle funzioni delle macromolecole, della logica costruttiva delle strutture biologiche e dei diversi livelli di organizzazione della materia vivente. Sono poi trattate la struttura e l’organizzazione funzionale delle cellule procarioti, con le caratteristiche dei domini Archea e Bacteria, e di quelle eucarioti, fino ai virus, responsabili di diverse malattie nelle piante negli animali e nell’uomo. Particolare risalto è dato ovviamente alla biologia della cellula eucariote, con una panoramica essenziale dei principi unitari che ne assicurano il bilancio energetico, e dei meccanismi alla base dell’espressione dell’informazione genetica. Sono quindi descritti i sistemi di membrana, le interazioni delle cellule con l’ambiente che le circonda, il ciclo cellulare e le divisioni delle cellule somatiche e germinali, la riproduzione sessuata e asessuata con cenni dei processi cellulari che guidano lo sviluppo iniziale dell’embrione di mammifero e umano, ed è dato particolare rilievo agli aspetti strutturali e fisiologici delle cellule muscolari striate e lisce e dell’apparato muscolare scheletrico/cardiaco. Sono infine trattati gli argomenti della genetica, con la struttura dei genomi procarioti ed eucarioti, la trasmissione, le mutazioni e l’evoluzione dell’informazione genetica, compresi i meccanismi, più recentemente identificati, delle modificazioni epigenetiche. Il testo così strutturato, sintetico ma esauriente, potrà soddisfare le esigenze di un’ampia fascia di studenti indipendentemente dal corso di studio prescelto. Con questo volume, tutti gli studenti saranno in grado di comprendere i principi biologici di base e il linguaggio appropriato, entrambi necessari allo studio dei corsi successivi, ma anche di identificare le aree di interesse, selezionare le fonti di informazione scientifica e interpretarne i dati, costruendosi un bagaglio culturale necessario sia alla formazione limitata al corso di laurea triennale, che all’eventuale proseguimento degli studi in corsi di laurea magistrale o di formazione post-laurea. Ringraziamo sentitamente il Dott. Piccin per la fiducia e l’incoraggiamento profusi nella realizzazione del manuale; la Sig.ra Barbara Cariali e il Sig. Marco Marzola per aver dato veste grafica al volume. Saremo inoltre molto grati a tutti coloro che vogliano segnalarci ogni errore, inesattezza o svista che, nonostante i nostri sforzi, il manuale possa contenere

Oxidative Stress Assessment in Response to Ultraendurance Exercise: Thiols Redox Status and ROS Production according to Duration of a Competitive Race

Purpose. Response to an ultraendurance competitive race on thiols redox status, reactive oxygen species (ROS) production, and oxidative stress (OxS) was investigated according to duration. Methods. Twenty-four elite runners were examined: six completed 50 km and eighteen 100 km. Blood and urine samples were collected before and immediately after the race. Erythrocytes and plasma aminothiols by high-performance liquid chromatography, total antioxidant capacity (TAC), and OxS biomarkers (protein carbonyl (PC), thiobarbituric acid-reactive substances (TBARS), 8-isoprostane (8-iso-PGF2α), and 8-OH-2-deoxyguanosine (8-OH-dG)) by immunoenzymatic assays and ROS production by Electron Paramagnetic Resonance were assessed. Results. Significant increases (P between <0.05 and <0.0001) were recorded in plasma total and oxidized aminothiols concentration and TAC (P<0.0001) only after 100 km: plasmatic (ROS production (+12 versus +29%), PC (+54 versus +115%), and TBARS (+28 versus +55%)) and urinary (8-OH-dG.creatinine−1 (+71 versus +158%) and 8-iso-PGF2α.creatinine−1 (+43 versus +135%)) concentrations for 50 and 100 km (duration 4 h 3′ versus 8 h 42′), respectively. Conclusion. Very prolonged ultraendurance exercise causes an increase in ROS production and OxS depending on specific biomarker examined but always linearly and directly related to exercise duration. Redox status of erythrocytes was preserved. A relationship between running performance and both prerace ROS production and antioxidant-redox status was found in 100 km race

R(+)-Thioctic Acid Effects on Oxidative Stress and Peripheral Neuropathy in Type II Diabetic Patients: Preliminary Results by Electron Paramagnetic Resonance and Electroneurography

Objectives. Diabetic neuropathy is the most common complication of diabetes. The idea of alterations in energy metabolism in diabetes is emerging. The biogenic antioxidant R(+)-thioctic acid has been successfully used in the treatment of diabetic polyneuropathic (DPN) patients. Methods. The effects of R(+)-thioctic acid (1 tablet, 1.6 g) administration were evaluated in 12 DPN patients at baseline and at 15, 30, 60, and 120 administration days throughout the assessment of oxidative stress (OxS); ROS production rate by electron paramagnetic resonance (EPR) technique; and oxidative damage biomarkers (thiobarbituric acid reactive substances (TBARS) and protein carbonyls (PC)), electroneurography (ENG) and visual analogue scale. Results. Supplementation induced significant changes (p<0.05) at 30 and 60 days. ROS production rate up to −16%; TBARS (−31%), PC (−38%), and TAC up to +48%. Motor nerve conduction velocity in SPE and ulnar nerves (+22% and +16%) and sensor conduction velocity in sural and median nerves (+22% and +5%). Patients reported a general wellness sensation improvement (+35%) at 30 days: lower limb pain sensation (−40%) and upper limbs (−23%). Conclusion. The results strongly indicate that an increased antioxidant capacity plays an important role in OxS, nerve conduction velocity, pain, and general wellness improvement. Nevertheless, the effects of the antioxidant compound were found positive up to 60 days. Then, a hormesis effect was observed. Novelty of the research would be a challenge for investigators to carefully address issues, including dose range factors, appropriate administration time, and targeting population to counteract possible “boomerang effects.” The great number of monitored parameters would firmly stress these conclusions

The “ON-OFF” Switching Response of Reactive Oxygen Species in Acute Normobaric Hypoxia: Preliminary Outcome

Exposure to acute normobaric hypoxia (NH) elicits reactive oxygen species (ROS) accumulation, whose production kinetics and oxidative damage were here investigated. Nine subjects were monitored while breathing an NH mixture (0.125 FIO2 in air, about 4100 m) and during recovery with room air. ROS production was assessed by Electron Paramagnetic Resonance in capillary blood. Total antioxidant capacity, lipid peroxidation (TBARS and 8-iso-PFG2α), protein oxidation (PC) and DNA oxidation (8-OH-dG) were measured in plasma and/or urine. The ROS production rate (μmol·min−1) was monitored (5, 15, 30, 60, 120, 240 and 300 min). A production peak (+50%) was reached at 4 h. The on-transient kinetics, exponentially fitted (t1/2 = 30 min r2 = 0.995), were ascribable to the low O2 tension transition and the mirror-like related SpO2 decrease: 15 min: −12%; 60 min: −18%. The exposure did not seem to affect the prooxidant/antioxidant balance. Significant increases in PC (+88%) and 8-OH-dG (+67%) at 4 h in TBARS (+33%) one hour after hypoxia offset were also observed. General malaise was described by most of the subjects. Under acute NH, ROS production and oxidative damage resulted in time and SpO2-dependent reversible phenomena. The experimental model could be suitable for evaluating the acclimatation level, a key element in the context of mountain rescues in relation to technical/medical workers who have not had enough time for acclimatization—as, for example, during helicopter flights

Acute Effects of Triathlon Race on Oxidative Stress Biomarkers

The response to strenuous exercise was investigated by reactive oxygen species (ROS) production, oxidative damage, thiol redox status, and inflammation assessments in 32 enrolled triathlon athletes (41.9±7.9 yrs) during Ironman® (IR), or half Ironman® (HIR) competition. In biological samples, inflammatory cytokines, aminothiols (glutathione (GSH), homocysteine (Hcy), cysteine (Cys), and cysteinylglycine (CysGly)), creatinine and neopterin, oxidative stress (OxS) biomarkers (protein carbonyl (PC), thiobarbituric acid-reactive substances (TBARS)), and ROS were assessed. Thirteen HIR and fourteen IR athletes finished the race. Postrace, ROS (HIR +20%; IR +28%; p<0.0001), TBARS (HIR +57%; IR +101%), PC (HIR +101%; IR +130%) and urinary neopterin (HIR +19%, IR +27%) significantly (range p<0.05-0.0001) increased. Moreover, HIR showed an increase in total Cys +28%, while IR showed total aminothiols, Cys, Hcy, CysGly, and GSH increase by +48, +30, +58, and +158%, respectively (range p<0.05-0.0001). ROS production was significantly correlated with TBARS and PC (R2=0.38 and R2=0.40; p<0.0001) and aminothiols levels (range R2=0.17-0.47; range p<0.01-0.0001). In particular, ROS was directly correlated with the athletes’ age (R2=0.19; p<0.05), with ultraendurance years of training (R2=0.18; p<0.05) and the days/week training activity (R2=0.16; p<0.05). Finally, the days/week training activity (hours/in the last 2 weeks) was found inversely correlated with the IL-6 postrace (R2=‐0.21; p<0.01). A strenuous performance, the Ironman® distance triathlon competition, alters the oxidant/antioxidant balance through a great OxS response that is directly correlated to the inflammatory parameters; furthermore, the obtained data suggest that an appropriate training time has to be selected in order to achieve the lowest ROS production and IL-6 concentration at the same time