Methodological pipeline for monitoring post-harvest quality of leafy vegetables.

Plants are primary source of nutrients for humans. However, the nutritional value of vegetables tends to decrease once organ and tissue sinks are detached from the plant. Minimal processing of leafy vegetables involves cutting and washing before packaging and storage. These processing procedures result in stressful conditions and post-harvest disorders senescence-related can also occur. The aim of this work is to define a methodological pipeline to evaluate the "quality" changes of fresh cut leafy vegetables over their shelf-life. At this purpose, intra-species variability has been investigated considering two varieties of Lactuca sativa (var. longifolia and capitata), showing different susceptibility to browning. Since browning mainly depends on phenol oxidation, redox parameters as well as the activity of the enzymes involved in phenol biosynthesis and oxidation have been monitored over storage time. At the same time, the metabolic changes of the lettuce leaves have been estimated as response patterns to chemical sensors. The obtained sensor outputs were predictive of browning-related biological features in a cultivar-dependent manner. The integration of the results obtained by this multivariate methodological approach allowed the identification of the most appropriate quality markers in lettuce leaves from different varieties. This methodological pipeline is proposed for the identification and subsequent monitoring of post-harvest quality of leafy vegetables.

Exploring the role of MKK7 in excitotoxicity and cerebral ischemia: a novel pharmacological strategy against brain injury.

Excitotoxicity following cerebral ischemia elicits a molecular cascade, which leads to neuronal death. c-Jun-N-terminal kinase (JNK) has a key role in excitotoxic cell death. We have previously shown that JNK inhibition by a specific cell-permeable peptide significantly reduces infarct size and neuronal death in an in vivo model of cerebral ischemia. However, systemic inhibition of JNK may have detrimental side effects, owing to blockade of its physiological function. Here we designed a new inhibitor peptide (growth arrest and DNA damage-inducible 45ß (GADD45ß-I)) targeting mitogen-activated protein kinase kinase 7 (MKK7), an upstream activator of JNK, which exclusively mediates JNK's pathological activation. GADD45ß-I was engineered by optimizing the domain of the GADD45ß, able to bind to MKK7, and by linking it to the TAT peptide sequence, to allow penetration of biological membranes. Our data clearly indicate that GADD45ß-I significantly reduces neuronal death in excitotoxicity induced by either N-methyl-D-aspartate exposure or by oxygen-glucose deprivation in vitro. Moreover, GADD45ß-I exerted neuroprotection in vivo in two models of ischemia, obtained by electrocoagulation and by thromboembolic occlusion of the middle cerebral artery (MCAo). Indeed, GADD45ß-I reduced the infarct size when injected 30 min before the lesion in both models. The peptide was also effective when administrated 6 h after lesion, as demonstrated in the electrocoagulation model. The neuroprotective effect of GADD45ß-I is long lasting; in fact, 1 week after MCAo the infarct volume was still reduced by 49%. Targeting MKK7 could represent a new therapeutic strategy for the treatment of ischemia and other pathologies involving MKK7/JNK activation. Moreover, this new inhibitor can be useful to further dissect the physiological and pathological role of the JNK pathway in the brain.