A novel assay for phosphoserine phosphatase exploiting serine acetyltransferase as the coupling enzyme

Phosphoserine phosphatase (PSP) catalyzes the final step of de novo L-serine biosynthesis— the hydrolysis of phosphoserine to serine and inorganic phosphate—in humans, bacteria, and plants. In published works, the reaction is typically monitored through the discontinuous malachite green phosphate assay or, more rarely, through a continuous assay that couples phosphate release to the phosphorolysis of a chromogenic nucleoside by the enzyme purine nucleoside phosphorylase (PNP). These assays suffer from numerous drawbacks, and both rely on the detection of phosphate. We describe a new continuous assay that monitors the release of serine by exploiting bacterial serine acetyltransferase (SAT) as a reporter enzyme. SAT acetylates serine, consuming acetyl-CoA and releasing CoA-SH. CoA-SH spontaneously reacts with Ellman’s reagent to produce a chromophore that absorbs light at 412 nm. The catalytic parameters estimated through the SAT-coupled assay are fully consistent with those obtained with the published methods, but the new assay exhibits several advantages. Particularly, it depletes L-serine, thus allowing more prolonged linearity in the kinetics. Moreover, as the SAT-coupled assay does not rely on phosphate detection, it can be used to investigate the inhibitory effect of phosphate on PSP

The NMDA receptor activation by D-serine and glycine is controlled by an astrocytic Phgdh-dependent serine shuttle

Astrocytes express the 3-phosphoglycerate dehydrogenase (Phgdh) enzyme required for the synthesis of L-serine from glucose. Astrocytic L-serine was proposed to regulate NMDAR activity by shuttling to neurons to sustain D-serine production, but this hypothesis remains untested. We now report that inhibition of astrocytic Phgdh suppressed the de novo synthesis of L-and D-serine and reduced the NMDAR synaptic potentials and long-term potentiation (LTP) at the Schaffer collaterals-CA1 synapse. Likewise, enzymatic removal of extracellular L-serine impaired LTP, supporting an L-serine shuttle mechanism between glia and neurons in generating the NMDAR coagonist D-serine. Moreover, deletion of serine racemase (SR) in glutamatergic neurons abrogated D-serine synthesis to the same extent as Phgdh inhibition, suggesting that neurons are the predominant source of the newly synthesized D-serine. We also found that the synaptic NMDAR activation in adult SR-knockout (KO) mice requires Phgdh-derived glycine, despite the sharp decline in the postnatal glycine levels as a result of the emergence of the glycine cleavage system. Unexpectedly, we also discovered that glycine regulates D-serine metabolism by a dual mechanism. The first consists of tonic inhibition of SR by intracellular glycine observed in vitro, primary cultures, and in vivo microdialysis. The second involves a transient glycine-induce D-serine release through the Asc-1 transporter, an effect abolished in Asc-1 KO mice and diminished by deleting SR in glutamatergic neurons. Our observations suggest that glycine is a multifaceted regulator of D-serine metabolism and implicate both D-serine and glycine in mediating NMDAR synaptic activation at the mature hippocampus through a Phgdh-dependent shuttle mechanism

The impact of multidisciplinary team management on outcome of hepatic resection in liver-limited colorectal metastases

Hepatic resection is the gold standard treatment for patients affected by liver-limited colorectal metastases. Reports addressing the impact of multidisciplinary team (MDT) evaluation on survival are controversial. The aim of this study was to evaluate the benefit of MDT management in these patients in our Institution experience. The objective of the analysis was to compare survivals of patients managed within our MDT (MDT cohort) to those of patients referred to surgery from other hospitals without MDT discussion (non-MDT cohort). Of the 523 patients, 229 were included in the MDT cohort and 294 in the non-MDT cohort. No difference between the two groups was found in terms of median overall survival (52.5 vs 53.6 months; HR 1.13; 95% CI, 0.88–1.45; p = 0.344). In the MDT cohort there was a higher number of metastases (4.5 vs 2.7; p < 0.0001). The median duration of chemotherapy was lower in MDT patients (8 vs 10 cycles; p < 0.001). Post-operative morbidity was lower in the MDT cohort (6.2 vs 21.5%; p < 0.001). One hundred and ninety-seven patients in each group were matched by propensity score and no significant difference was observed between the two groups in terms of OS and DFS. Our study does not demonstrate a survival benefit from MDT management, but it allows surgery to patients with a more advanced disease. MDT assessment reduces the median duration of chemotherapy and post-operative morbidities

Erratum to: Drivers for energy efficiency and their effect on barriers: empirical evidence from Italian manufacturing enterprises (Energy Efficiency, (2017), 10, 4, (855-869), 10.1007/s12053-016-9488-x)

© Springer Science+Business Media Dordrecht 2016. This paper was unfortunately published with error. The given name of one of the authors was incorrectly captured as “Vincenzo” when it should be “Giovanni.” Further, the email address should be corrected to [email protected]. This was then corrected and is not anymore present in the published copies of the article

Classification of drivers for industrial energy efficiency and their effect on the barriers affecting the investment decision-making process

© 2016, Springer Science+Business Media Dordrecht. Industrial energy efficiency represents a priority for European industrial competitiveness. Many studies offer contributions providing evidence of the existence of driving forces supporting the adoption of energy efficiency measures, but a structured approach to drivers for industrial energy efficiency is still lacking. Therefore, in the present study, we propose a definition of drivers, making emphasis on the industrial decision-maker perspective, that is needed for their classification here proposed. Focus is given to point out the difference between internal and external drivers, highlighting the major stakeholders responsible for their promotion. Drivers are further categorized into: regulatory, economic, informative, and vocational training. Moreover, we propose a framework describing the effect of drivers on barriers in the decision-making process, as well as a preliminary identification of the major stakeholders to promote drivers. The study opens several opportunities for further research in the area of industrial energy efficiency

Drivers for energy efficiency and their effect on barriers: empirical evidence from Italian manufacturing enterprises

© 2017, Springer Science+Business Media Dordrecht. Industrial activities are responsible for a significant share of both global delivered energy demand and CO2 emissions. Hence, a widespread adoption of energy-efficient technologies and practices represents a crucial means for sustainable production. Adopting a novel framework of drivers for energy efficiency describing the effect of drivers on barriers in the decision-making process steps and able to account for the nature of drivers and the stakeholders responsible for their promotion, we have performed an exploratory investigation into 61 manufacturing small- and medium-sized enterprises in Northern Italy. Our findings have highlighted the importance of information and economic drivers, showing the need for enterprises to be supported not only by public institutions but also by external stakeholders involved in the supply of energy-efficient technologies and practices such as industrial associations and groupings, as well as service and technology suppliers. Moreover, our study has highlighted an almost equal relevance of both internal and external drivers. According to our findings, the increase of awareness—generating the interest to energy issues and stimulated both by external and internal stakeholders—as well as financial issues have emerged as the most critical in the decision-making process to adopt an energy-efficient measure. A preliminary comprehension of the mechanisms relating drivers to barriers in the decision-making process brought additional value to the study, highlighting the most effective and specific means to overcome the existing barriers. We have also explored the effect of several firm characteristics, such as firm size and energy intensity offering suggestions for industrial decision-makers as well as policy-makers