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In this study, the effects of repeated cycles of drying and rehydration on some physiological traits were assessed in long shelf-life tomatoes cultivated in a typical semi-arid area of Southern Italy. Three Sicilian landraces (‘Custonaci’, ‘Salina’, and ‘Vulcano’) from the germplasm collection at CNR-IBE (Catania, Italy) and a commercial tomato mini-plum (‘Faino Hy., control) were investigated under three water regimes: DRY (no irrigation), IRR (long-season full irrigation) and REW (post-drought rewaterings). Net photosynthetic assimilation rate (Pn), leaf transpiration (E), stomatal conductance (gs), instantaneous water use efficiency (WUEi), leaf intercellular CO2 (Ci, ppm), and leaf temperature (°C), were measured during the growing season. At harvest (late July), fruit production per plant was measured and ripened fruits were analysed for total solids (TS), soluble solids (SS), reducing sugars (RS), vitamin C (AscA), and total phenols (TP). Pn promptly responded to rewatering (REW), quickly increasing immediately after irrigation, and declined with soil drying up. All genotypes had similar physiological pathways in DRY, but in IRR, ‘Faino’ had higher Pn (up to 31 μmol CO2 m−2s−1) and E (up to 18 mmol H2O m−2s−1). Stomatal conductance (gs) after rewatering steeply increased and quickly declined after that. All local landraces had the same gs in IRR and REW. Variations in RWC were less pronounced than those in other physiological parameters. WUEi in REW and DRY proceeded similarly (up to 3 μmol CO2 mmol H2O). Irrigation in REW significantly promoted plant productivity over the DRY control (up to +150% in ‘Vulcano’). TS and SS in REW were lower than those in DRY, but higher (+19 and +7%, respectively) than in IRR. Vitamin C was greater in DRY and REW (26 and 18% higher than in IRR, respectively). TP in all local tomatoes were significantly higher (up to +29% in ‘Vulcano’) than those in the commercial control. Water regime had a minor effect on TP in ‘Custonaci’ and ‘Salina’. Principal Component Analysis (PCA) provided information on the changes in physiological and fruit quality traits in tomatoes in relation to cultivars and water regimes. The results of this study also revealed that a water-saving irrigation strategy where few irrigations are applied after prolonged periods of drought might be profitable in terms of fruit production enhancement in long shelf-life tomatoes and that limited rewaterings in most cases, help retaining high levels of fruit quality traits.

Summary: Endometriosis is a benign condition characterized by the presence of ectopic endometrial tissue. Our study investigated the effect of endometriosis on the risk of endometrial cancer (EC) and the prognosis of endometriosis-associated EC. In our study, 197,196 patients with endometriosis and without a previous diagnosis of EC were compared with 6,455,556 females encountering health services for examinations, with body mass index (BMI) data, and without endometriosis or EC. A propensity score generated 197,141 matched pairs. In the endometriosis cohort, 875 cases of EC were seen, whereas 558 were in the control group: the hazard ratio (HR) was 1.56 (95% CI 1.40–1.73, p

Abstract Castor (Ricinus communis L.) is an important oilseed crop worldwide whose inedible oil is widely used in the industrial, pharmaceutical, and agricultural sectors. Castor plants show high conversion potential for use as biorefining feedstocks. The present study was conducted to investigate the effects of two nitrogen fertilization levels (0 and 120 kg N ha−1) on seed and oil yield. From a biorefinery perspective, the residual biomass of seed processing was analyzed in terms of fiber composition and biomethane production carrying out a biological pretreatment using two white‐rot fungi (Pleurotus ostreatus and Irpex lacteus). Nitrogen fertilization resulted in an increase in seed and oil yields and a difference in capsule husk composition. Fungal pretreatment of capsule husks showed promising effects on anaerobic digestion, increasing the biomethane yield compared to untreated biomass. The highest lignin degradation and the lowest cellulose loss during pretreatment were obtained with I. lacteus, and this fungal pretreatment resulted in the highest biomethane yield (103.2 NmL g−1 volatile solids) for the fertilized biomass.

Abstract Background The effects of awake prone position on the breathing pattern of hypoxemic patients need to be better understood. We conducted a crossover trial to assess the physiological effects of awake prone position in patients with acute hypoxemic respiratory failure. Methods Fifteen patients with acute hypoxemic respiratory failure and PaO2/FiO2 0.99) and ΔP L (9 [7–11] cmH2O vs. 8 [5–9], p = 0.17). Airway resistance and time constant were higher in prone vs. supine position (9 cmH2O s arbitrary units−3 [4–11] vs. 6 [4–9], p = 0.05; 0.53 s [0.32–61] vs. 0.40 [0.37–0.44], p = 0.03). Prone position increased EELI (3887 arbitrary units [3414–8547] vs. 1456 [959–2420], p = 0.002) and promoted V T distribution towards dorsal lung regions without affecting V T size and lung compliance: this generated lower dynamic strain (0.21 [0.16–0.24] vs. 0.38 [0.30–0.49], p = 0.004). The magnitude of pendelluft phenomenon was not different between study phases (55% [7–57] of V T in prone vs. 31% [14–55] in supine position, p > 0.99). Conclusions Prone position improves oxygenation, increases EELI and promotes V T distribution towards dependent lung regions without affecting V T size, ΔP L, lung compliance and pendelluft magnitude. Prone position reduces respiratory rate and increases ΔP ES because of positional increases in airway resistance and prolonged expiratory time. Because high ΔP ES is the main mechanistic determinant of self-inflicted lung injury, caution may be needed in using awake prone position in patients exhibiting intense ΔP ES. Clinical trail registeration: The study was registered on clinicaltrials.gov (NCT03095300) on March 29, 2017.

Abstract Background Long-term outcomes of patients treated with helmet noninvasive ventilation (NIV) are unknown: safety concerns regarding the risk of patient self-inflicted lung injury and delayed intubation exist when NIV is applied in hypoxemic patients. We assessed the 6-month outcome of patients who received helmet NIV or high-flow nasal oxygen for COVID-19 hypoxemic respiratory failure. Methods In this prespecified analysis of a randomized trial of helmet NIV versus high-flow nasal oxygen (HENIVOT), clinical status, physical performance (6-min-walking-test and 30-s chair stand test), respiratory function and quality of life (EuroQoL five dimensions five levels questionnaire, EuroQoL VAS, SF36 and Post-Traumatic Stress Disorder Checklist for the DSM) were evaluated 6 months after the enrollment. Results Among 80 patients who were alive, 71 (89%) completed the follow-up: 35 had received helmet NIV, 36 high-flow oxygen. There was no inter-group difference in any item concerning vital signs (N = 4), physical performance (N = 18), respiratory function (N = 27), quality of life (N = 21) and laboratory tests (N = 15). Arthralgia was significantly lower in the helmet group (16% vs. 55%, p = 0.002). Fifty-two percent of patients in helmet group vs. 63% of patients in high-flow group had diffusing capacity of the lungs for carbon monoxide

Abstract Background Continuous measurement of urinary PO2 (PuO2) is being applied to indirectly monitor renal medullary PO2. However, when applied to critically ill patients with shock, its measurement may be affected by changes in FiO2 and PaO2 and potential associated O2 diffusion between urine and ureteric or bladder tissue. We aimed to investigate PuO2 measurements in septic shock patients with a fiberoptic luminescence optode inserted into the urinary catheter lumen in relation to episodes of FiO2 change. We also evaluated medullary and urinary oxygen tension values in Merino ewes at two different FiO2 levels. Results In 10 human patients, there were 32 FiO2 decreases and 31 increases in FiO2. Median pre-decrease FiO2 was 0.36 [0.30, 0.39] and median post-decrease FiO2 was 0.30 [0.23, 0.30], p = 0.006. PaO2 levels decreased from 83 mmHg [77, 94] to 72 [62, 80] mmHg, p = 0.009. However, PuO2 was 23.2 mmHg [20.5, 29.0] before and 24.2 mmHg [20.6, 26.3] after the intervention (p = 0.56). The median pre-increase FiO2 was 0.30 [0.21, 0.30] and median post-increase FiO2 was 0.35 [0.30, 0.40], p = 0.008. PaO2 levels increased from 64 mmHg [58, 72 mmHg] to 71 mmHg [70, 100], p = 0.04. However, PuO2 was 25.0 mmHg [IQR: 20.7, 26.8] before and 24.3 mmHg [IQR: 20.7, 26.3] after the intervention (p = 0.65). A mixed linear regression model showed a weak correlation between the variation in PaO2 and the variation in PuO2 values. In 9 Merino ewes, when comparing oxygen tension levels between FiO2 of 0.21 and 0.40, medullary values did not differ (25.1 ± 13.4 mmHg vs. 27.9 ± 15.4 mmHg, respectively, p = 0.6766) and this was similar to urinary oxygen values (27.1 ± 6.17 mmHg vs. 29.7 ± 4.41 mmHg, respectively, p = 0.3192). Conclusions Changes in FiO2 and PaO2 within the context of usual care did not affect PuO2. Our findings were supported by experimental data and suggest that PuO2 can be used as biomarker of medullary oxygenation irrespective of FiO2.

Abstract Introduction Helmet noninvasive support may provide advantages over other noninvasive oxygenation strategies in the management of acute hypoxemic respiratory failure. In this narrative review based on a systematic search of the literature, we summarize the rationale, mechanism of action and technicalities for helmet support in hypoxemic patients. Main results In hypoxemic patients, helmet can facilitate noninvasive application of continuous positive-airway pressure or pressure-support ventilation via a hood interface that seals at the neck and is secured by straps under the arms. Helmet use requires specific settings. Continuous positive-airway pressure is delivered through a high-flow generator or a Venturi system connected to the inspiratory port of the interface, and a positive end-expiratory pressure valve place at the expiratory port of the helmet; alternatively, pressure-support ventilation is delivered by connecting the helmet to a mechanical ventilator through a bi-tube circuit. The helmet interface allows continuous treatments with high positive end-expiratory pressure with good patient comfort. Preliminary data suggest that helmet noninvasive ventilation (NIV) may provide physiological benefits compared to other noninvasive oxygenation strategies (conventional oxygen, facemask NIV, high-flow nasal oxygen) in non-hypercapnic patients with moderate-to-severe hypoxemia (PaO2/FiO2 ≤ 200 mmHg), possibly because higher positive end-expiratory pressure (10–15 cmH2O) can be applied for prolonged periods with good tolerability. This improves oxygenation, limits ventilator inhomogeneities, and may attenuate the potential harm of lung and diaphragm injury caused by vigorous inspiratory effort. The potential superiority of helmet support for reducing the risk of intubation has been hypothesized in small, pilot randomized trials and in a network metanalysis. Conclusions Helmet noninvasive support represents a promising tool for the initial management of patients with severe hypoxemic respiratory failure. Currently, the lack of confidence with this and technique and the absence of conclusive data regarding its efficacy render helmet use limited to specific settings, with expert and trained personnel. As per other noninvasive oxygenation strategies, careful clinical and physiological monitoring during the treatment is essential to early identify treatment failure and avoid delays in intubation.

ABSTRACT Medication‐related osteonecrosis of the jaw (MRONJ) is a rare but serious adverse effect of antiresorptive medications administered for control of osseous malignancy, osteoporosis, or other bone metabolic diseases. Despite being reported in the literature two decades ago, MRONJ etiology, pathophysiology, and progression remain largely unknown, and current nonoperative or operative treatment strategies are mostly empirical. Several hypotheses that attempt to explain the mechanisms of MRONJ pathogenesis have been proposed. However, none of these hypotheses alone is able to capture the complex mechanistic underpinnings of the disease. In this minireview, we aim to highlight key findings from clinical and translational studies and propose a unifying model for the pathogenesis and progression of MRONJ. We also identify aspects of the disease process that require further investigation and suggest areas for future research efforts toward calibrating methodologic approaches and validating experimental findings. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

Abstract Growing industrial crops on marginal lands has been proposed as a strategy to minimize competition for arable land and food production. In the present study, eight experimental sites in three different climatic zones in Europe (Mediterranean, Atlantic and Continental), seven advanced industrial crop species [giant reed (two clones), miscanthus (M. × giganteus and two new seed‐based hybrids), saccharum (one clones), switchgrass (one variety), tall wheatgrass (one variety), industrial hemp (three varieties) and willow (eleven clones)], and six marginality factors alone or in combination (dryness, unfavorable texture, stoniness, shallow soil, topsoil acidity, heavy metal and metalloid contamination) were investigated. At each site, biophysical constraints and low‐input management practices were combined with prevailing climatic conditions. The relative yield of a site‐specific low‐input system compared with the site‐specific control was from small to large (i.e. from −99% in industrial hemp in the Mediterranean to +210% in willow in the Continental zone), due to the genotype‐by‐management interaction along with climatic variation between growing seasons. Genotype selection and improved knowledge on crop response to changing environmental, site‐specific biophysical constraint and input application has been detected as key to profitably grow industrial crops on marginal areas. This study may act to provide hints on how to scale up investigated cropping systems, through low‐input practices, under similar environmental and soil conditions tested at each site. However, further attention to detail on the agronomy of early plant development and management in larger multi‐year and multi‐location field studies with commercially scalable agronomies are needed to validate yield performances, and thereby to inform on the best industrial crop options.

The aim of the present work was to assess the leaf area duration (LAD) and the radiation use efficiency (RUE) of six warm-season perennial biomass grasses (PBGs) in a two-year field trial in the semiarid Mediterranean climate under different soil water availability. Two ecotypes of giant reed (Arundo donax L., ARCT and ARMO), one ecotype of African fodder cane (Saccharum spontaneum L. subsp. aegyptiacum (Willd.) Hack., SAC) and three hybrids of Miscanthus (the commercial M. × giganteus J.M. Greef, Deuter ex Hodk., Renvoize, M × G, and two new seed-based hybrids, GNT9 and GNT10) were compared under three levels of soil water availability: rainfed, 50% and 100% of maximum crop evapotranspiration (ETm) restoration. The determination of RUE of perennial plants is controversial and has led to contrasting results in past studies. In the present work, LAD and RUE differed among crops and irrigation regimes, being positively affected by supplemental water inputs. SAC, ARCT and ARMO showed both high LAD and RUE, which determined the high biomass yield than both the commercial M × G and the improved Miscanthus hybrids GNT9 and GNT10. RUE was particularly high and less affected by soil water availability during the mid-season, while the effect of irrigation and the differences among the genotypes were larger during the late season. Adequate biomass yield can be achieved by sub-optimal soil water availability, thus reducing the water footprint and increasing the sustainability of these biomass perennial grasses selected for the Mediterranean climate.

To date, economically and energy-costly vegetative propagation using rhizomes and tissue culture are the only options for the cultivation of Miscanthus spp. Some genotypes of miscanthus produce fertile seeds, offering a valid alternative to vegetative propagation. A preliminary study has been conducted on the seeds of two hybrids of miscanthus obtained via interspecific cross-pollination between M. sacchariflorus and M. sinensis: ‘GRC14’ (maternal: M. sacchariflorus) and ‘GRC10B’ (maternal: M. sinensis). Seeds were assessed for germination traits in a laboratory (at 25 °C in the dark) just after panicle harvest, and during 1-year storage at room temperature or at 8 °C. In a second experiment, the effects of gibberellic acid (GA3) solution at different concentrations (0, 50, 100, 300, 500 ppm) on the germination of freshly matured seeds were assessed. Poor germination just after harvest (3, but the extent of the GA effect was genotype-dependent. In conclusion, a low establishment rate may result from direct seeding when fresh seed is used in the field. In this case, the use of GA3 is a possible strategy to ameliorate the impact of dormancy on seed germination. In the case of delayed sowings in late winter–early spring, seeds stored at room temperature after harvest may better perform than those stored at 8 °C.

Castor (Ricinus communis L.) is an oilseed species that can be grown as a semi-perennial in Mediterranean environments, including the coastal areas of Sicily. The present study investigated the optimization of cultivation techniques for castor, with the crop being maintained over a two-year period, through the evaluation of different agronomical inputs in order to increase seed yield. The effects of irrigation (I) and nitrogen fertilization (N) on the seed and oil yield and their components were assessed in castor cultivated in a typical semi-arid environment. Four levels of irrigation (I0, I30, I60, and I100: 0, 30, 60, and 100% of crop evapotranspiration—ETm restoration, respectively) as the main plot and three levels of nitrogen fertilization (N) (0, 60, and 120 kg N ha−1) as the sub-plot were considered. Irrigation mostly affected the number of racemes per plant, the number of capsules per raceme, and the seed weight. The oil content was, on average, 39.2% and 45.6% for the first and second year, respectively. The highest seed yield was obtained by I100N120 treatment (4154.0 kg ha−1); however, the combination of a high soil nitrogen level (N120) and medium water availability (I60) resulted in satisfactory seed and oil yields. The reduction in the irrigation water to an intermediate level could be also an environmentally friendly strategy not significantly affecting yields.

In this study, the allelochemical-induced effects of the leaf water extract of hemp (Cannabis sativa) on seed germination and seedling growth were investigated in durum wheat (Triticum durum) cv. Mongibello and barley (Hordeum vulgare) cv. Alamo. The seeds were imbibed in 25%, 50%, 75%, or 100% solution concentrations, obtained from a 2% hemp leaf extract, and they were germinated at 20 and 10 °C. The 2% extract stock solution (i.e., 100% solution concentration) was obtained by extracting 20 g of leaf powder in 1 L distilled water (2%, w/v). A control of seeds imbibed in distilled water was considered. Germination was scored daily and at the end of the germination test, the final germination percentage (FGP), mean germination time (MGT), germination index (GI), and vigor index (VI) were calculated. The radicles and shoots of the germinated seeds of both species were also measured for their length and weight. The analyses revealed the presence of phenols in the hemp leaf extract, which were the highest in the 100% concentration (13.14 ± 0.20 mg gallic acid equivalent-GAE g−1 extract). At 20 °C, increasing the concentration had either minor (in wheat) or no effects (in barley) on the seed germination. At 10 °C, the germination was reduced by the increasing concentration, suggesting a potential allelopathic effect of the extract. However, at 100% concentration, the seeds germinated significantly better that those at 75% concentration. At 20 °C, increasing concentration progressively depressed both the radicle and shoot elongation in both species. At 10 °C, the radicle elongation was depressed by the extract, irrespective of the concentration, whilst the shoots were as long as those of the control in wheat, and smaller at the 50 and 75% concentrations in barley. We can conclude that the water extract of hemp leaves can have a negative impact on wheat and barley seed germination and seedling growth, particularly at temperatures below optimal. However, the high concentration of phenols in the extract may stimulate seed germination and seedling growth. Thus, in semi-arid areas, such as those of the Mediterranean regions, sowings of wheat and barley after hemp at an optimal (i.e., 20 °C) or nearly optimal temperature are recommended in order to minimize the combined stressing effects of the temperature and allelopathic leaf extract upon plant establishment, which may limit a successful crop establishment.

Some physiological, oxidative, and antioxidant enzymatic patterns were assessed in plants of three local Sicilian landraces of long shelf-life tomatoes (‘Custonaci’, ‘Salina’, and ‘Vulcano’), as compared to the commercial tomato hybrid ‘Faino’ (control). Three water treatments were considered in open-field: DRY (no irrigation); IRR (long-season full irrigation); REW (drought/rewatering cycles). During the growing season, soil water deficit (SWD) was estimated, and relative water content (RWC), specific leaf area (SLA), proline and malondialdehyde (MDA) content, and glutathione peroxidase (GPX), catalase (CAT), and superoxide dismutase (SOD) activities were measured in leaves. Differently from control, RWC in local landraces exhibited a similar pattern in REW and DRY, indicating a low capacity to re-hydrate after rewatering. Positive correlation of proline content vs. SWD in all local landraces highlights an osmotic adjustment occurring in these tomatoes in response to limited soil water content. Long shelf-life tomatoes suffered minor oxidative stress due to severe soil water deficit, as revealed by the lower levels of MDA with respect to the control. Significant correlation of CAT vs. SWD for all tomatoes indicates that this antioxidant enzyme, among those analyzed, may be considered as a biomarker for a water stress condition more than for oxidative stress due to water deficit.

Vitamin D covers roles of paramount importance in the regulation of multiple physiological pathways of the organism. The metabolism of vitamin D involves kidney–liver crosstalk and requires an adequate function of these organs, where vitamin D is progressively turned into active forms. Vitamin D deficiency has been widely reported in patients living in the community, being prevalent among the most vulnerable subjects. It has been also documented in many critically ill patients upon admission to the intensive care unit. In this context, vitamin D deficiency may represent a risk factor for the development of life-threatening clinical conditions (e.g., infection and sepsis) and worse clinical outcomes. Several researchers have investigated the impact of vitamin D supplementation showing its feasibility, safety, and effectiveness, although conflicting results have put into question its real benefit in critically ill patients. The existing studies included heterogeneous critically ill populations and used slightly different protocols of vitamin D supplementation. For these reasons, pooling up the results is difficult and not conclusive. In this narrative review, we described vitamin D physiology and the pathophysiology of vitamin D depletion with a specific focus on critically ill patients with liver dysfunction, acute kidney injury, acute respiratory failure, and sepsis.

Feedback inhibition by horizontal cells regulates rod and cone photoreceptor calcium channels that control their release of the neurotransmitter glutamate. This inhibition contributes to synaptic gain control and the formation of the center-surround antagonistic receptive fields passed on to all downstream neurons, which is important for contrast sensitivity and color opponency in vision. In contrast to the plasmalemmal GABA transporter found in non-mammalian horizontal cells, there is evidence that the mechanism by which mammalian horizontal cells inhibit photoreceptors involves the vesicular release of the inhibitory neurotransmitter GABA. Historically, inconsistent findings of GABA and its biosynthetic enzyme, L-glutamate decarboxylase (GAD) in horizontal cells, and the apparent lack of surround response block by GABAergic agents diminished support for GABA's role in feedback inhibition. However, the immunolocalization of the vesicular GABA transporter (VGAT) in the dendritic and axonal endings of horizontal cells that innervate photoreceptor terminals suggested GABA was released via vesicular exocytosis. To test the idea that GABA is released from vesicles, we localized GABA and GAD, multiple SNARE complex proteins, synaptic vesicle proteins, and Cav channels that mediate exocytosis to horizontal cell dendritic tips and axonal terminals. To address the perceived relative paucity of synaptic vesicles in horizontal cell endings, we used conical electron tomography on mouse and guinea pig retinas that revealed small, clear-core vesicles, along with a few clathrin-coated vesicles and endosomes in horizontal cell processes within photoreceptor terminals. Some small-diameter vesicles were adjacent to the plasma membrane and plasma membrane specializations. To assess vesicular release, a functional assay involving incubation of retinal slices in luminal VGAT-C antibodies demonstrated vesicles fused with the membrane in a depolarization- and calcium-dependent manner, and these labeled vesicles can fuse multiple times. Finally, targeted elimination of VGAT in horizontal cells resulted in a loss of tonic, autaptic GABA currents, and of inhibitory feedback modulation of the cone photoreceptor Cai, consistent with the elimination of GABA release from horizontal cell endings. These results in mammalian retina identify the central role of vesicular release of GABA from horizontal cells in the feedback inhibition of photoreceptors.

Abstract Background Rhabdomyosarcomas (RMSs) are the most frequent soft tissue sarcoma in children and adolescents, defined by skeletal muscle differentiation and the status of FOXO1 fusions. In pediatric malignancies, in particular RMS, scant and controversial observations are reported about PD-L1 expression as a putative biomarker and few immune checkpoint clinical trials are conducted. Methods PD-L1 assessment was evaluated by immunohistochemistry (IHC) utilizing two anti-PDL1 antibodies, in a pilot cohort of 25 RMS. Results were confirmed in primary and commercial RMS cell lines by cytofluorimetric analysis and IHC. Results PD-L1 expression was detectable, by both anti-PD-L1 antibodies, in the immune contexture of immune cells infiltrating and/or surrounding the tumor, in 15/25 (60%) RMS, while absent expression was observed in neoplastic cells. Flow cytometry analysis and PD-L1 IHC of commercial and primary RMS cell lines confirmed a very small percentage of PD-L1 positive-tumor cells, under the detection limits of conventional IHC. Interestingly, increased PD-L1 expression was observed in the immune contexture of 4 RMS cases post chemotherapy compared to their matched pre-treatment samples. Conclusion Here we identify a peculiar pattern of PD-L1 expression in our RMS series with scanty positive-tumor cells detected by flow cytometry, and recurrent expression in the immune cells surrounding or infiltrating the tumor burden.

Endocytosis is a fundamental process for internalizing material from the plasma membrane, including many transmembrane proteins that are selectively internalized depending on environmental conditions. In most cells, the main route of entry is clathrin-mediated endocytosis (CME), a process that involves the coordinated activity of over 60 proteins; however, there are likely as-yet unidentified proteins involved in cargo selection and/or regulation of endocytosis. We performed a mutagenic screen to identify novel endocytic genes in Saccharomyces cerevisiae expressing the methionine permease Mup1 tagged with pHluorin (pHl), a pH-sensitive GFP variant whose fluorescence is quenched upon delivery to the acidic vacuole lumen. We used fluorescence-activated cell sorting to isolate mutagenized cells with elevated fluorescence, resulting from failure to traffic Mup1-pHl cargo to the vacuole, and further assessed subcellular localization of Mup1-pHl to characterize the endocytic defects in 256 mutants. A subset of mutant strains was classified as having general endocytic defects based on mislocalization of additional cargo proteins. Within this group, we identified mutations in four genes encoding proteins with known roles in endocytosis: the endocytic coat components SLA2, SLA1, and EDE1, and the ARP3 gene, whose product is involved in nucleating actin filaments to form branched networks. All four mutants demonstrated aberrant dynamics of the endocytic machinery at sites of CME; moreover, the arp3R346H mutation showed reduced actin nucleation activity in vitro. Finally, whole genome sequencing of two general endocytic mutants identified mutations in conserved genes not previously implicated in endocytosis, KRE33 and IQG1, demonstrating that our screening approach can be used to identify new components involved in endocytosis.

The Mediterranean long shelf-life tomato (or long storage tomato) is a plant traditionally cultivated under no irrigation in the Southern regions of Italy, whose fruits have typical high sensory and nutritional quality. However, yield levels are kept low under current cultivation conditions. In this study, the effects of repeated cycles of drying and rehydration on crop productivity and nutritional quality of fruits in terms of polyphenols and carotenoids content were assessed in long shelf-life tomatoes cultivated in a typical semi-arid area of Southern Italy. The three local Sicilian landraces ‘Custonaci’, ‘Salina’ and ‘Vulcano’, and the commercial tomato hybrid ‘Faino’ (control) were submitted to three irrigation treatments: DRY (no irrigation); IRR (long-season full irrigation); REW (drought/rewatering cycles). Total 450, 4710, and 1849 m3 ha−1, were distributed in DRY, IRR, and REW, respectively. At harvest, fruit yield, polyphenols (as total, flavonoids, and hydroxycinnamoyl quinic acids-HCQA), and carotenoids contents (lycopene and β-carotene) were measured. All cultivars benefitted from very limited irrigation in REW, raising their productivity (up to +147% in ‘Vulcano’) with respect to that of plants overstressed in DRY. Irrigation water use efficiency in REW was higher than that in IRR. Water shortage in REW led to a polyphenols content of fruits that was slightly lower (171.1 μg g−1) than that in DRY, but higher than that in IRR (116.8 μg g−1). All local landraces had greater contents (>160 μg g−1) than control (113.0 μg g−1). Under REW and DRY, the two landraces ‘Salina’ and ‘Vulcano’ produced fruits with the same polyphenols and flavonoids contents. Overall, the two water stressed treatments (DRY and REW) did not differ for HCQA content (>66 μg g−1), which was significantly higher than that in the irrigated plots (−1). Lycopene content in DRY and REW exceeded 100 μg g−1; among local types, ‘Vulcano’ exhibited the highest content (106.7 μg g−1). The β-carotene content did not change in ‘Custonaci’ and ‘Salina’, whereas in ‘Vulcano’ it was greater in DRY and REW. Only in ‘Faino’ water limitation in REW exerted a clear promoting effect on β-carotene content of fruits. The results of the present study reveal how rewatering after long-lasting drought periods is beneficial to long shelf-life tomatoes in terms of productivity. Appropriate drought cycles in REW may also help the fruits to retain high nutritional levels.

Neurodegenerative retinal diseases, such as glaucoma and diabetic retinopathy, involve a gradual loss of neurons in the retina as the disease progresses. Central nervous system neurons are not able to regenerate in mammals, therefore, an often sought after course of treatment for neuronal loss follows a neuroprotective or regenerative strategy. Neuroprotection is the process of preserving the structure and function of the neurons that have survived a harmful insult; while regenerative approaches aim to replace or rewire the neurons and synaptic connections that were lost, or induce regrowth of damaged axons or dendrites. In order to test the neuroprotective effectiveness or the regenerative capacity of a particular agent, a robust experimental model of retinal neuronal damage is essential. Zebrafish are being used more often in this type of study because their eye structure and development is well-conserved between zebrafish and mammals. Zebrafish are robust genetic tools and are relatively inexpensive to maintain. The large array of functional and behavioral tests available in zebrafish makes them an attractive model for neuroprotection studies. Some common insults used to model retinal disease and study neuroprotection in zebrafish include intense light, chemical toxicity and mechanical damage. This review covers the existing retinal neuroprotection and regeneration literature in the zebrafish and highlights their potential for future studies.

Within the project “BIOSEA” funded by the Italian Ministry of Agriculture and Forestry, a preliminary laboratory test was conducted to assess the variability for cold tolerance during germination in 30 cultivars of biomass sorghum, among fiber and sweet types. Seed germination (%) and mean germination time (MGT) were examined at seven constant temperatures (from 8 °C to 35 °C) and base temperature (Tb) and thermal time (θT) for 50% germination were calculated. A wide genetic diversity in the germination response of sorghum was ascertained at 8 °C (CV 45%) and 10 °C (CV 25.4%). At 8 °C, in cultivars of ‘Padana 4’, ‘PR811F’, ‘PSE24213’, ‘PR849’ and ‘Zerberus’, seed germination exceeded 80%. Seeds of ‘Zerberus’ were also the fastest, requiring less than 13 days for final germination at this low temperature. Great differences were found in Tb and θT among cultivars. Tb varied between 7.44 °C (‘PR811F’) and 13.48 °C (‘Nectar’). Thermal time (θT) was, on average, 24.09 °Cd−1, and ranged between 16.62 (‘Nectar’) and 33.42 °Cd−1 (‘PSE24213’). The best combination of the two germination parameters (i.e., low Tb and θT) corresponded to ‘Zerberus’, ‘Sucrosorgo 506’, ‘Jumbo’ and ‘PR811F’. Accordingly, these cultivars are more tolerant to cold stress during germination and, thus, more adapt to early spring sowings in Mediterranean areas (March-April). Cultivars ‘PR811F’ (fiber type) and ‘Sucrosorgo 506’ (sweet type) also combine high cold tolerance with good productivity in terms of final dry biomass, as assessed in open-field conditions (late spring sowing). The genetic variation in the germination response to a low temperature is useful for the identification of genotypes of sorghum suitable to early sowings in semi-arid areas. Selection within existing cultivars for cold tolerance during germination may also contribute to the expansion of biomass sorghum into cooler cultivation areas, such as those of Northern Europe, which are less suitable to this warm season crop.

The solar photosphere and the outer layer of the Sun’s interior are characterized by convective motions, which display a chaotic and turbulent character. In this work, we evaluated the pseudo-Lyapunov exponents of the overshooting convective motions observed on the Sun’s surface by using a method employed in the literature to estimate those exponents, as well as another technique deduced from their definition. We analyzed observations taken with state-of-the-art instruments at ground- and space-based telescopes, and we particularly benefited from the spectro-polarimetric data acquired with the Interferometric Bidimensional Spectrometer, the Crisp Imaging SpectroPolarimeter, and the Helioseismic and Magnetic Imager. Following previous studies in the literature, we computed maps of four quantities which were representative of the physical properties of solar plasma in each observation, and estimated the pseudo-Lyapunov exponents from the residuals between the values of the quantities computed at any point in the map and the mean of values over the whole map. In contrast to previous results reported in the literature, we found that the computed exponents hold negative values, which are typical of a dissipative regime, for all the quantities derived from our observations. The values of the estimated exponents increase with the spatial resolution of the data and are almost unaffected by small concentrations of magnetic field. Finally, we showed that similar results were also achieved by estimating the exponents from residuals between the values at each point in maps derived from observations taken at different times. The latter estimation technique better accounts for the definition of these exponents than the method employed in previous studies.

In Mediterranean environments, few perennial grass species are available for cultivation in rainfed systems and marginal lands, where plants with excellent adaptation are required. The aim of the present work was to determine the potentiality of five native Mediterranean perennial grasses for lignocellulosic biomass production. Wild accessions of three hemicryptophytes (Ampelodesmos mauritanicus, Hyparrhenia hirta, and Piptatherum miliaceum) and two geophytes (Saccharum spontaneum ssp. aegyptiacum and Sorghum halepense) were collected at three Mediterranean sites (Sicily, Sardinia and Majorca), and their morphological, physiological, productivity and quality traits were evaluated in the field. The species differed in height, with S. spontaneum and A. mauritanicus being the tallest. The leaf mass ratio ranged from 0.23 to 1.0 g g–1 among species. Maximum net photosynthesis was measured in the C4 species S. spontaneum and S. halepense (26.6 and 23.8 mmol CO2 m–2 s–1, respectively). A. mauritanicus showed the lowest transpiration rate and the highest instantaneous water use efficiency (2.7 mmol H2O m–2 s–1 and 6.9 mmol CO2 mmol H2O–1, respectively). S. spontaneum was the most productive species, yielding more than 18 Mg DM ha–1 as a three-year average. The highest content of acid detergent lignin was found in P. miliaceum, while A. mauritanicus was the species richest in hemicellulose and cellulose and poorest in ash. S. spontaneum showed the highest moisture content at harvest. Overall, the studied species showed interesting morphological, physiological, productive and qualitative traits. Nevertheless, additional research is necessary to investigate their long-term performance under different management strategies.

Medullary carcinoma is a rare malignant tumor of the kidney. It affects individuals of African descent and all cases reported show evidence of sickle cell trait. We reviewed an unusual carcinoma arising in a white man, the ninth in the literature. The tumor demonstrated features associated with renal medullary carcinoma, or unclassified renal cell carcinoma, medullary phenotype as recently described; the presence of sickle cell trait confirmed the diagnosis of medullary carcinoma. This case is helpful in the differential diagnosis with non-sickle cell associated “renal cell carcinoma, unclassified with medullary phenotype,” and study of this spectrum of tumors is ongoing.

In the present study, a two-year field trial was carried out with the aim to evaluate daylength and air temperature effects on leaf appearance and related rates in two durum wheat (Triticum durum Desf.), two bread wheat (Triticum aestivum L.) and two barley (Hordeum vulgare L.) cultivars, using six different sowing dates (SD). Significant effects of SD on final main stem leaf number (FLN), thermal leaf appearance rate (TLAR), daily leaf appearance rate (DLAR) and phyllochron (PhL) were found. Cultivars resulted inversely correlated to mean air temperature in the interval emergence - fifth leaf full expansion (E-V). Linear response of leaf number over days after sowing was shown for all SD and cultivars, with R2 higher than 0.95. FLN linearly decreased from the first to the last SD for durum wheat, while more variable behaviour was observed in bread wheat. TLAR and DLAR showed a linear increment of the rate from the first to the last SD in durum wheat, while did not for bread wheat and barley. PhL in durum wheat decreased from the first to the last SD. Barley and bread wheat showed the highest values on those SDs which did not reach flowering. The increase of TLAR was affected by photoperiod and photothermal units in durum wheat, while by temperatures only in barley and bread wheat. Present results might find practical application in the improvement of phenology simulation models for durum wheat, bread wheat and barley grown in Mediterranean area in absence of water and nutrient stress.

The function of dimethylsulphoniopropionate (DMSP) in plants is unclear. It has been proposed as an antioxidant, osmolyte and overflow for excess energy under stress conditions. The formation of DMSP is part of the methionine (MET) pathway that is involved in plant stress responses. We used a new analytical approach to accurately quantify the changes in DMSP concentration that occurred in two ecotypes of the biomass crop Arundo donax subject to moderate drought stress under field conditions. The ecotypes of A. donax were from a hot semi-arid habitat in Morocco and a warm-humid environment in Central Italy. The Moroccan ecotype showed more pronounced reductions in photosynthesis, stomatal conductance and photochemical electron transport than the Italian ecotype. An increase in isoprene emission occurred in both ecotypes alongside enhanced foliar concentrations of DMSP, indicative of a protective function of these two metabolites in the amelioration of the deleterious effects of excess energy and oxidative stress. This is consistent with the modification of carbon within the methyl-erythritol and MET pathways responsible for increased synthesis of isoprene and DMSP under moderate drought. The results of this study indicate that DMSP is an important adaptive component of the stress response regulated via the MET pathway in A. donax. DMSP is likely a multifunctional molecule playing a number of roles in the response of A. donax to reduced water availability.

Great effort has been placed to identify the most suited bioenergy crop under different environments and management practices, however, there is still need to find new genetic resources for constrained areas. For instance, South Mediterranean area is strongly affected by prolonged drought, high vapour pressure deficit (VPD) and extremely high temperatures during summertime. In the present work we investigated the soil water effect on crop growth and leaf gas exchange of Saccharum spontaneum L. ssp. aegyptiacum (Willd.) Hackel, a perennial, rhizomatous, herbaceous grass. Furthermore, the net increase of biomass production per unit light intercepted [radiation use efficiency (RUE)] and per unit water transpired [water use efficiency (WUE)] was also studied. To this end a field trial was carried out imposing three levels of soil water availability (I100, I50 and I0, corresponding to 100%, 50% and 0% of ETm restutition) under a semi-arid Mediterranean environment. Leaf area index (LAI), stem height, biomass dry matter yield, CO2 assimilation rate, and transpiration rate resulted significantly affected by measurement time and irrigation treatment, with the highest values in I100 and the lowest in I0. RUE was the highest in I100 followed by I50 and I0; on the other hand, WUE was higher in I0 than I50 and I100. At LAI values greater than 2.0, 85% photosynthetically active radiation was intercepted by the Saccharum stand, irrespective of the irrigation treatment. Saccharum spontaneum spp. aegyptiacum is a potential species for biomass production in environment characterized by drought stress, high temperatures and high VPD, as those of Southern Europe and similar semi-arid areas.

In the Iblei plateau (Sicily, Southern Italy) the native dairy cattle breed Modicana during the spring season grazes exclusively on natural pastures for the production of the Ragusano protected denomination of origin cheese. Along the grazing season, herbage undergoes to changes on protein, fibre and moisture content, affecting quality parameters such as plant carotenoids concentration, involved in the colour and nutritional characteristics of dairy products and potential biomarkers for authenticating fed green pasture-based diets. The aim of this work was to assess whether the cheese traceability index, based on the carotenoids spectra data elaboration, could be related to seasonal variations of floral composition and pasture quality. Four herbage and cheese samples were collected every two weeks in two representative farms of this area, from March to May 2013. Pasture characteristics as pastoral vegetation composition and pastoral value were analysed using the methodology developed for pastoral resources studies. Traceability index showed a significant positive correlation with pasture moisture and crude protein content (r=0.729* and 0.853**, respectively), while it was negatively correlated with fibre content (r=–0.719*).

The review, after a short introduction on the tendencies of the European Community Policy on biomasses, describes the agronomic, energy potential and environmental aspects of biomass crops for energy in relation to the research activity carried out in Italy on this topic, differentiating crops on the basis of the main energy use: biodiesel and bioethanol (which refers to “first generation biofuel”), heat and electricity. Currently, many of the crops for potential energy purposes are food crops (wheat, barley, corn, rapeseed, soybean, sunflower, grain sorghum, sugar beet) and their production may be used as biofuel source (bioethanol and biodiesel) since their crop management aspects are well known and consequently they are immediately applicable. Other species that could be used, highly productive in biomass, such as herbaceous perennial crops (Arundo donax, Miscanthus spp., cardoon), annual crops (sweet sorghum), short rotation woody crops (SRF) have been carefully considered in Italy, but they still exhibit critical aspects related to propagation technique, low-input response, harvest and storage technique, cultivars and mechanization. Crops for food, however, often have negative energetic indices and environmental impacts (carbon sequestration, Life Cycle Assessment), consequent to their low productivity. Conversely, crops which are more productive in biomass, show both a more favourable energy balance and environmental impact.

In this paper the suitability of three perennial, herbaceous, lignocellulosic grasses (Arundo donax, Saccharum spontaneous spp. aegyptiacum and Miscanthus x giganteus) for the production of second-generation bioethanol in semi-arid Mediterranean environment was studied. Crops were established in spring 2002, supplying irrigation and nitrogen fertilization up to 2004/2005 growing season. Subsequently, crops were grown without any agronomic input and harvested annually. Data reported in this paper refers to 2008/2009 and 2009/2010 growing seasons. Aboveground dry matter (DM) yield was higher in Arundo (35.4±2.1 Mg ha–1 in 2009 and 32.2±1.9 Mg ha–1 in 2010 harvest) than in Saccharum (27.3±2.0 and 23.9±1.9 Mg ha–1, respectively) and Miscanthus (19.6±2.8 and 17.2±1.6 Mg ha–1, respectively). Structural polysaccharides of the raw material were higher in Miscanthus (63.4% w/w) followed by Saccharum (61.5% w/w) and Arundo (57.6% w/w). The same trend was identified for the cellulose content (41.0%, 36.8% and 34.6%, respectively). The highest values in the total hemicellulose complex were observed in Saccharum (24.7%), followed by Arundo (23.1%) and Miscanthus (22.4%). The composition of structural polysaccharides leads to a higher theoretical ethanol yield (TEY) from one dry ton of Miscanthus feedstock (kg DM Mg–1), followed by Saccharum and Arundo. On the other hand, the TEY per unit surface (Mg ha–1) was greater in Arundo than in Saccharum and Miscanthus. When compared to other lignocellulosic sources used in the second-generation bioethanol technology, such as agricultural residues, woody species and other herbaceous perennial crops, Arundo, Saccharum and Miscanthus showed a great potential in terms of TEY ha–1. Given the high levels of biomass yield and composition of structural polysaccharides, the three species might be introduced into the Mediterranean cropping systems to supply lignocellulosic biomass for second-generation industrial plants or bio-refineries.

Bioenergy crops are expected to play an important role in reducing CO2 emission, in energy supply and in European energy policy. However, a sustainable bioenergy supply must be resilient to climate change and the impacts on agriculture at both global and regional scale. The purpose of this study was to forecast the potential distribution of several bioenergy crops based on agronomic and environmental constrains under current conditions and future scenarios (2020 and 2030) in European Union. Potential biomass yield, according to the category end use product achievable in each environmental zone of Europe at present and in the future available land have been also studied. Future yields were assessed according to two factors: technological development and climate change: the former was based on prospect of DG-Agriculture for conventional crops and expert judgments for bioenergy crops, while the latter based on relevant research papers and literature reviews which used site-specific crop growth models. Yields are expected to increase in northern Europe due to climate change and technological development, while in southerneastern Europe the negative effect of climate change will be mitigated by the technological development. The estimated total biomass production in Europe, on the basis of future yields and surplus land made available for energy crops, may not be sufficient to meet the needs of bioenergy supply as claimed in the European directive 2009/28/EC.

Enterohemorrhagic Escherichia coli (EHEC) is dependent on acid resistance for gastric passage and low oral infectious dose, and the locus of enterocyte effacement (LEE) for intestinal colonization. Mutation of rpoN, encoding sigma factor N (σ(N)), dramatically alters the growth-phase dependent regulation of both acid resistance and the LEE. This study reports on the determinants of σ(N)-directed acid resistance and LEE expression, and the underlying mechanism attributable to this phenotype. Glutamate-dependent acid resistance (GDAR) in TW14359ΔrpoN correlated with increased expression of the gadX-gadW regulatory circuit during exponential growth, whereas upregulation of arginine-dependent acid resistance (ADAR) genes adiA and adiC in TW14359ΔrpoN did not confer acid resistance by the ADAR mechanism. LEE regulatory (ler), structural (espA and cesT) and effector (tir) genes were downregulated in TW14359ΔrpoN, and mutation of rpoS encoding sigma factor 38 (σ(S)) in TW14359ΔrpoN restored acid resistance and LEE genes to WT levels. Stability, but not the absolute level, of σ(S) was increased in TW14359ΔrpoN; however, increased stability was not solely attributable to the GDAR and LEE expression phenotype. Complementation of TW14359ΔrpoN with a σ(N) allele that binds RNA polymerase (RNAP) but not DNA, did not restore WT levels of σ(S) stability, gadE, ler or GDAR, indicating a dependence on transcription from a σ(N) promoter(s) and not RNAP competition for the phenotype. Among a library of σ(N) enhancer binding protein mutants, only TW14359ΔntrC, inactivated for nitrogen regulatory protein NtrC, phenocopied TW14359ΔrpoN for σ(S) stability, GDAR and ler expression. The results of this study suggest that during exponential growth, NtrC-σ(N) regulate GDAR and LEE expression through downregulation of σ(S) at the post-translational level; likely by altering σ(S) stability or activity. The regulatory interplay between NtrC, other EBPs, and σ(N)-σ(S), represents a mechanism by which EHEC can coordinate GDAR, LEE expression and other cellular functions, with nitrogen availability and physiologic stimuli.

Photoreceptor terminals contain post-synaptic density (PSD) proteins e.g., PSD-95/PSD-93, but their role at photoreceptor synapses is not known. PSDs are generally restricted to post-synaptic boutons in central neurons and form scaffolding with multiple proteins that have structural and functional roles in neuronal signaling. The Shank family of proteins (Shank 1-3) functions as putative anchoring proteins for PSDs and is involved in the organization of cytoskeletal/signaling complexes in neurons. Specifically, Shank 1 is restricted to neurons and interacts with both receptors and signaling molecules at central neurons to regulate plasticity. However, it is not known whether Shank 1 is expressed at photoreceptor terminals. In this study we have investigated Shank 1A localization in the outer retina at photoreceptor terminals. We find that Shank 1A is expressed presynaptically in cone pedicles, but not rod spherules, and it is absent from mice in which the Shank 1 gene is deleted. Shank 1A co-localizes with PSD-95, peanut agglutinin, a marker of cone terminals, and glycogen phosphorylase, a cone specific marker. These findings provide convincing evidence for Shank 1A expression in both the inner and outer plexiform layers, and indicate a potential role for PSD-95/Shank 1 complexes at cone synapses in the outer retina.

BACKGROUND:The global disparity in cancer incidence remains a major public health problem. We focused on prostate cancer since microscopic disease in men is common, but the incidence of clinical disease varies more than 100 fold worldwide. Ca(2+) signaling is a central regulator of cell proliferation, but has received little attention in cancer prevention. We and others have reported a strong dose-dependent reduction in the incidence of prostate and lung cancer within populations exposed to boron (B) in drinking water and food; and in tumor and cell proliferation in animal and cell culture models. METHODS/PRINCIPAL FINDINGS:We examined the impact of B on Ca(2+) stores using cancer and non-cancer human prostate cell lines, Ca(2+) indicators Rhod-2 AM and Indo-1 AM and confocal microscopy. In DU-145 cells, inhibition of Ca(2+) release was apparent following treatment with Ringers containing RyR agonists cADPR, 4CmC or caffeine and respective levels of BA (50 microM), (1, 10 microM) or (10, 20, 50,150 microM). Less aggressive LNCaP cancer cells required 20 microM BA and the non-tumor cell line PWR1E required 150 microM BA to significantly inhibit caffeine stimulated Ca(2+) release. BA (10 microM) and the RyR antagonist dantroline (10 microM) were equivalent in their ability to inhibit ER Ca(2+) loss. Flow cytometry and confocal microscopy analysis showed exposure of DU-145 cells to 50 microM BA for 1 hr decreased stored [Ca(2+)] by 32%. CONCLUSION/SIGNIFICANCE:We show B causes a dose dependent decrease of Ca(2+) release from ryanodine receptor sensitive stores. This occurred at BA concentrations present in blood of geographically disparate populations. Our results suggest higher BA blood levels lower the risk of prostate cancer by reducing intracellular Ca(2+) signals and storage.