Biochemical and immunochemical similarities among mammalian bilitranslocase and a plant flavonoid translocator

Flavonoids are a large class of plant secondary metabolites, belonging to polyphenol family, which possess pharmacological and nutritional properties. Their synthesis takes place only in plants, while mammals can acquire them only with diet. It has been demonstrated that flavonoid uptake occurs in rat also by the activity of bilitranslocase, a carrier that is involved in anion transport in liver cell, vascular endothelium and gastric mucosa. A sequence of bilitranslocase interacting with flavonoid moieties is already known and characterized. Antibody raised against such protein epitope were shown to exhibit cross-reactivity against plant membrane proteins in tissues involved in flavonoid transport and accumulation, such as teguments of carnation petals and skin of grape berries. Further immunolocalization studies allowed to demonstrate the presence of cross-reacting protein not only at the level of tegumental tissues, but also associated to sieve elements and seed teguments in grape berries

Off-Adherence Keeping (OAK) observational study: intentional off-adherence immunomodulatory multiple sclerosis treatment

Aims: To evaluate how improved treatment adherence with a lower-frequency regimen/treatment of intramuscular (IM) IFN beta-1a impacts therapeutic effectiveness in relapsing-remitting multiple sclerosis (MS) patients switching from a higher-frequency injectable regimen/treatment. Patients & methods: Italian patients with relapsing-remitting MS and prior poor adherence to high-frequency injectable treatments (n = 181) were followed for 24 months after starting IM IFN beta-1a. Results: During the study, 97.4% of patients were treatment adherent; 22.1% of patients reported a relapse. The estimated probability of remaining relapse-free after 2 years was 78%. A high dropout rate (52.5%) led to small sample size and reduced statistical power. Conclusion: Intramuscular IFN beta-1a treatment was associated with high adherence and a low relapse rate. Unfortunately, low patient retention limited the generalizability of these findings.Plain language summary: Prior research suggests that taking the drug IFN beta-1a through less frequent muscle injections enables more patients to adhere to their prescription than taking other medications. This study included 181 Italian patients with relapsing-remitting multiple sclerosis (MS) who historically did not take medication as often as prescribed. Relapses of MS were counted among patients treated with muscle injections of IFN beta-1a for 2 years; 97.4% of patients followed their prescription and 22.1% experienced a relapse. From these data, 78% of patients were estimated not to experience a relapse during 2 years of IFN beta-1a muscle injections. However, an unusually high number of patients (52.5%) left the study within 2 years, which makes it difficult to draw firm conclusions

Flavonoids and darkness lower PCD in senescing Vitis vinifera suspension cell cultures

Background Senescence is a key developmental process occurring during the life cycle of plants that can be induced also by environmental conditions, such as starvation and/or darkness. During senescence, strict control of genes regulates ordered degradation and dismantling events, the most remarkable of which are genetically programmed cell death (PCD) and, in most cases, an upregulation of flavonoid biosynthesis in the presence of light. Flavonoids are secondary metabolites that play multiple essential roles in development, reproduction and defence of plants, partly due to their well-known antioxidant properties, which could affect also the same cell death machinery. To understand further the effect of endogenously-produced flavonoids and their interplay with different environment (light or dark) conditions, two portions (red and green) of a senescing grapevine callus were used to obtain suspension cell cultures. Red Suspension cell Cultures (RSC) and Green Suspension cell Cultures (GSC) were finally grown under either dark or light conditions for 6 days. Results Darkness enhanced cell death (mainly necrosis) in suspension cell culture, when compared to those grown under light condition. Furthermore, RSC with high flavonoid content showed a higher viability compared to GSC and were more protected toward PCD, in accordance to their high content in flavonoids, which might quench ROS, thus limiting the relative signalling cascade. Conversely, PCD was mainly occurring in GSC and further increased by light, as it was shown by cytochrome c release and TUNEL assays. Conclusions Endogenous flavonoids were shown to be good candidates for exploiting an efficient protection against oxidative stress and PCD induction. Light seemed to be an important environmental factor able to induce PCD, especially in GSC, which lacking of flavonoids were not capable of preventing oxidative damage and signalling leading to senescence

CHARACTERIZATION OF LIPASE ACTIVITY IN GREEN COFFEE BEANS DURING STORAGE AND GERMINATION

Lipase, green coffee, germination, storage, lipid Coffee seeds possess an intermediate storage pattern, showing a non-quiescent behaviour characterized by various metabolic reactions occurring during storage. In particular, lipase is the main enzyme involved in the mobilization of triacylglycerols, providing energy and a source of carbon skeleton during early stages of germination. During storage, the triacylglycerols might be involved in the generation of undesirable compounds (known as \u201coff-flavours\u201d), lowering both the viability of coffee seeds and the cup quality. In this work, the soluble protein fraction from coffee seeds and plantlets was extracted by acetone and then was utilised to assay lipase activity. Green coffee beans, harvested in Colombia, were stored at room temperature for 2-3 months, in order to verify the influence of prolonged storage on lipase activity. For germination experiments, the seeds were imbibed for 7 days at 30 \ub0C and transferred in perlite at 28 \ub0C and 90% R.H. for further 3 weeks. Lipase activity was detected by a colorimetric method based on specific degradation of a chromogenic substrate, at pH 8.2. Green coffee seeds exhibited an appreciable lipase activity that was slightly increased during storage. Such an activity was inhibited by tetrahydrolipstatin (THL) in a concentration-dependent manner, while it was slightly stimulated by both EGTA and EDTA. During the germination, after 10, 14, 17 and 21 days, lipase activity showed an initial increase that was followed by a gradual decrease. The effect of the presence or absence of the parchment (seed coat), during the first stages of germination, has also been investigated

Phospholipase activities in green coffee beans (Coffea arabica L.) harvested in different countries

Triacylglycerols (TAGs) are accumulated in specialised organelles called \u201coil bodies\u201d, which are enclosed in a phospholipid monolayer embedded with some unique proteins. Upon germination, such membranes are modified to allow the availability of TAGs as an energy source during early stages of seedling growth in oilseeds. This process occurs by the sequential and/or collective action of many hydrolytic enzymes, such as phospholipases, lipoxygenases and lipases that are associated to oil body membranes. In contrast, during seed storage, oilseed lipids may undergo lipolytic degradation processes leading to a wide range of metabolites potentially harmful for seed viability. In particular, green coffee endosperm consist of approx. 99% of the mature seed mass and contains many polyunsaturated fatty acids whose degradation leads to volatile compound formation through the oxylipin pathway. In spite of this, the enzymes involved in TAGs degradation (particularly lipases) are poorly studied. Therefore, the aim of this work was to evaluate the involvement of phospholipase activity in oil body membrane degradation during storage of green coffee (Coffea arabica L.) and to determine the correlations between storage lipid mobilization and maintenance of seed viability in beans harvested in different countries (Ethiopia, India, Kenya and Tanzania). Green coffee beans were frozen and powdered in liquid nitrogen and oil bodies were extracted with cold acetone. Phospholipase A1 and A2 activities were assayed in crude extracts by a fluorimetric method, using different probes. Such activities were just partially stimulated by free Ca2+, in contrast with what reported by others. Furthermore, PLA2 activity was assayed in a wide range of pH, evidencing two peaks of pH optimum. These results suggests that green coffee bean presents at least two isoforms of PLA2. Phospholipase profiles (PLA2 and total) were correlated with the provenience of the beans, showing a higher activity in those harvested in Ethiopia, while the lower was associated to beans from India

The permeability transition in plant mitochondria: The missing link

The synthesis of ATP in mitochondria is dependent on a low permeability of the inner membrane. Nevertheless, mitochondria can undergo an increased permeability to solutes, named permeability transition (PT) that is mediated by a pore (PTP). PTP opening requires matrix Ca2+ and leads to mitochondrial swelling and release of intramembrane space proteins (e.g. cytochrome c). This feature has been initially observed in mammalian mitochondria and tentatively attributed to some components present either in the outer or inner membrane. Recent works on mammalian mitochondria point to mitochondrial ATP synthase dimers as physical basis for PT, a finding that has been substantiated in yeast and Drosophila mitochondria. In plant mitochondria, swelling and release of proteins have been linked to programmed cell death, but in isolated mitochondria PT has been observed in only a few cases and in plant cell cultures only indirect evidence is available. The possibility that mitochondrial ATP synthase dimers could function as PTP also in plants is discussed here on the basis of the current evidence. Finally, a hypothetical explanation for the origin of PTP is provided in the framework of molecular exaptation

Cross Cultural Adaptation and Validation of Italian Version of the Leeds Assessment of Neuropathic Symptoms and Signs Scale and Pain DETECT Questionnaire for the Distinction between Nociceptive and Neuropathic Pain

Objective. This study aimed to validate Italian versions of Leeds Assessment of Neuropathic Symptoms and Signs (LANSS) scale and Pain DETECT questionnaire (PD-Q) and evaluate the ability of these questionnaires to discriminate between nociceptive and neuropathic pain. Design. Multicenter prospective validation cohort study. Subjects and Setting. One hundred patients were included with a diagnosis formulated by a specialist in outpatient settings (50 affected by knee osteoarthritis as nociceptive pain and 50 affected by trigeminal or postherpetic neuralgia as neuropathic pain). Methods. The Italian versions of both questionnaires according to Italian cultural characteristics were performed according to the following steps: (1) translation of the questionnaires from English into Italian; (2) review by a bilingual individual for consistency; (3) proposed version after a mail round between experts; (4) backward translation; (5) comparison with the original English version by the experts; (6) approved version of the questionnaires. One hundred patients were enrolled and completed the two questionnaires administered by a specialist or blinded nursing staff, at the baseline and after 24/48 hours. Internal consistency, stability, validity, and discriminative power were analyzed. Results. Statistically significant differences were reported about the ability of both questionnaires to discriminate between patients affected by neuropathic or nociceptive pain. Internal consistency for the Italian version of the LANSS was 0.76, and for PD-Q, it was 0.80, assessed by Cronbach’s α; LANSS showed a good test-retest reliability with an ICC of 0.76, and PD-Q showed a high test-retest reliability with an ICC of 0.96. For interrater reliability, there was a concordance rate of 83.3% between reference diagnosis and LANSS (Cohen’s kappa = 0.67, CI 95% 0.52–0.75). Conclusions. This study validated the Italian versions of LANSS and PD-Q as reliable instruments with good psychometric characteristics, for pain evaluation, discriminating between nociceptive and neuropathic pain. Our findings were similar to those observed in the original study. Furthermore, we have reported the test-retest reliability for both questionnaires, not addressed in original validation studies

Properties of the permeability transition of pea stem mitochondria

In striking analogy with Saccharomyces cerevisiae, etiolated pea stem mitochondria did not show appreciable Ca2+ uptake. Only treatment with the ionophore ETH129 (which allows electrophoretic Ca2+ equilibration) caused Ca2+ uptake followed by increased inner membrane permeability, membrane depolarization and Ca2+ release. Like the permeability transition (PT) of mammals, yeast and Drosophila, the PT of pea stem mitochondria was stimulated by diamide and phenylarsine oxide and inhibited by MgADP and Mg-ATP, suggesting a common underlying mechanism; yet, the plant PT also displayed distinctive features: (i) as in mammals it was desensitized by cyclosporin A, which does not affect the PT of yeast and Drosophila; (ii) similarly to S. cerevisiae and Drosophila it was inhibited by Pi, which stimulates the PT of mammals; (iii) like in mammals and Drosophila it was sensitized by benzodiazepine 423, which is ineffective in S. cerevisiae; (iv) like what observed in Drosophila it did not mediate swelling and cytochrome c release, which is instead seen in mammals and S. cerevisiae. We find that cyclophilin D, the mitochondrial receptor for cyclosporin A, is present in pea stem mitochondria. These results indicate that the plant PT has unique features and suggest that, as in Drosophila, it may provide pea stem mitochondria with a Ca2+ release channel

Pornography: Non-consensual, Vengeful, Online

Structural characterization of kaempferol standard by Mass spectra, using MSn. Upper spectrum shows full spectrum of kaempferol. Lower spectrum shows spectrum of CID of the [M-H]ˉ ion at m/z 285 ion of kaempferol. (TIF 32 kb

Do dimers of ATP synthase form the PTP in pea stem mitochondria?

In animal cells Ca2+ and ROS induce a sudden change in the inner mitochondrial membrane permeability, which has been named Permeability Transition (PT). Recently, it has been proposed that dimers of F-ATP synthase form the Permeability Transition Pore (PTP), the megachannel involved in this phenomenon [1]. This feature has not yet been characterized in plants, even if their mitochondria possess the candidate components for PTP formation. Therefore, wecharacterized the functional properties of PTP in plant mitochondria and verified if F-ATP synthase possesses channel function in electrophysiology experiments. Mitochondria isolated from pea stem underwent PT when Ca2+ was added in the presence of the ionophore ETH129. The membrane electrical potential was then collapsed and the phenomenon matched by Ca2+ release but not by mitochondrial swelling. As is observed with the PT of animal mitochondria, Cyclosporin A (CsA) significantly delayed occurrence of PT, which was inhibited by Mg2+-nucleotides and favored by benzodiazepine Bz-423 and oxidants, such as phenylarsine oxide and diamide. In electrophysiology experiments, F-ATP synthase dimers inserted into an artificial bilayer showed channel activity characterized by a rather small conductance, which could explain the inability of plant PTP to mediate mitochondrial swelling. These data suggest that F-ATP synthase is involved in PTP formation also in plant mitochondria