Corrigendum to "Synthesis and characterization of a (+)-catechin and L-(+)-ascorbic acid cocrystal as a new functional ingredient for tea drinks" [Heliyon 5 (8) (August 2019) e02291].

[This corrects the article DOI: 10.1016/j.heliyon.2019.e02291.].

Synthesis and characterization of a (+)-catechin and L-(+)-ascorbic acid cocrystal as a new functional ingredient for tea drinks.

Tea (Camellia Sinensis) is one of the most popular drink, consumed as infusion or bottled ready to drink beverages. Although tea leaves contain many antioxidants compounds, after processing they can drastically decrease, sometimes up to a full degradation, as in the case of catechin, a very healthy flavan-3-ol. In this context, the synthesis of a cocrystal between (+)-catechin and L-(+)-ascorbic acid, was proved to be a useful strategy to make a new ingredient able to ameliorate the antioxidant profile of both infusions and bottled teas. The obtained cocrystal showed a three-fold higher solubility than (+)catechin and its formation was elucidated unambiguously by FT-IR, thermal (DSC) and diffraction (PXRD) analyses. Antioxidant characteristics of the samples were evaluated by colorimetric assays. As expected, infusions showed much better antioxidant features than ready-to-use lemon and peach teas. The same trend was confirmed after the addition of the cocrystal at two concentration levels. In particular, supplementation at concentration of 2 mg mL-1 improved the bottled tea antioxidant values to the level showed by the not-added infusion tea.

The impact of cultivar on polyphenol and biogenic amine profiles in Calabrian red grapes during winemaking.

In this study, during winemaking, was evaluated the influence of cultivar on bioactive compounds (organic acids, d-(+)-glucose, d-(-)-fructose, biogenic amines (BAs), anthocyanins, polyphenols and flavonoids) and antioxidant activity of Calabrian (Southern Italy) autochthonous grapes (Arvino, Gaglioppo, Greco Nero, Magliocco Canino, Magliocco Dolce and Nocera). Phenolic compounds increased from grapes to wine for all varieties. Arvino grapevine showed the highest DPPH radical scavenging activity, while a promising inhibition of the lipid peroxidation was observed with Greco Nero grapes. BAs were mostly formed during alcoholic fermentation and Arvino always showed the lowest BAs amounts, while Magliocco Canino generally exhibited the highest. Collectively, the results demonstrated that Calabrian autochthonous grapevines were rich in sugars, organic acids and phenolic compounds thus allowing the production of high quality wines.

Immunohistochemical expression of stem cell markers in pheochromocytomas/paragangliomas is associated with SDHx mutations.

OBJECTIVE: Pheochromocytomas (PCCs) are neuroendocrine tumors that occur in the adrenal medulla, whereas paragangliomas (PGLs) arise from paraganglia in the head, neck, thorax, or abdomen. In a variety of tumors, cancer cells with stem cell-like properties seem to form the basis of tumor initiation because of their ability to self-renew and proliferate. Specifically targeting this small cell population may lay the foundation for more effective therapeutic approaches. In the present study, we intended to identify stem cells in PCCs/PGLs. DESIGN: We examined the immunohistochemical expression of 11 stem cell markers (SOX2, LIN28, NGFR, THY1, PREF1, SOX17, NESTIN, CD117, OCT3/4, NANOG, and CD133) on tissue microarrays containing 208 PCCs/PGLs with different genetic backgrounds from five European centers. RESULTS: SOX2, LIN28, NGFR, and THY1 were expressed in more than 10% of tumors, and PREF1, SOX17, NESTIN, and CD117 were expressed in <10% of the samples. OCT3/4, NANOG, and CD133 were not detectable at all. Double staining for chromogranin A/SOX2 and S100/SOX2 demonstrated SOX2 immunopositivity in both tumor and adjacent sustentacular cells. The expression of SOX2, SOX17, NGFR, LIN28, PREF1, and THY1 was significantly associated with mutations in one of the succinate dehydrogenase (SDH) genes. In addition, NGFR expression was significantly correlated with metastatic disease. CONCLUSION: Immunohistochemical expression of stem cell markers was found in a subset of PCCs/PGLs. Further studies are required to validate whether some stem cell-associated markers, such as SOX2, could serve as targets for therapeutic approaches and whether NGFR expression could be utilized as a predictor of malignancy.

PKB/AKT phosphorylation of the transcription factor Twist-1 at Ser42 inhibits p53 activity in response to DNA damage.

Protein kinase B (PKB/Akt) is ubiquitously expressed in cells. Phosphorylation of its multiple targets in response to various stimuli, including growth factors or cytokines, promotes cell survival and inhibits apoptosis. PKB is upregulated in many different cancers and a significant amount of the enzyme is present in its activated form. Here we show that PKB phosphorylates one of the anti-apoptotic proteins--transcription factor Twist-1 at Ser42. Cells expressing Twist-1 displayed inefficient p53 upregulation in response to DNA damage induced by gamma-irradiation or the genotoxic drug adriamycin. This influenced the activation of p53 target genes such as p21(Waf1) and Bax and led to aberrant cell-cycle regulation and the inhibition of apoptosis. The impaired induction of these p53 effector molecules is likely to be mediated by PKB-dependent phosphorylation of Twist-1 because, unlike the wild-type mutant, the Twist-1 S42A mutant did not confer cell resistance to DNA damage. Moreover, phosphorylation of Twist-1 at Ser42 was shown in vivo in various human cancer tissues, suggesting that this post-translational modification ensures functional activation of Twist-1 after promotion of survival during carcinogenesis.

Inhibitory effect of voluntary movement preparation on cutaneous heat pain and laser-evoked potentials.

In our study, preparation of voluntary movement was used to physiologically activate the motor cortex areas and the effect of this activation on CO(2) laser-evoked potentials (LEPs) was explored. LEPs were recorded from 31 scalp electrodes in 10 healthy subjects after painful stimulation of the right C6-C7 skin dermatomes. LEP stimuli were delivered in the time interval between a visual warning stimulus followed after 1 s. by an imperative stimulus. The imperative stimulus triggered: (i) no task in the baseline condition (Pain); (ii) flexion-extension movements of the second finger of the right hand in the movement condition (Pain + Movement); (iii) cognitive task (mathematic computation) in the distraction condition (Pain + Cognition). The experimental conditions were also repeated during application of laser stimuli on the left C6-C7 skin dermatomes. Compared with the baseline condition (no task required), during preparation of right-hand voluntary movement there was a significant reduction in LEP amplitude and subjective pain rating after right- but not after left-hand stimulation, which suggests that the observed effect cannot be attributed to a nonspecific reduction in attention toward painful stimulus. During preparation of a cognitive task, LEP amplitude was reduced compared to baseline. Our results represent the first neurophysiological suggestion that physiological activation of the motor cortex, occurring during movement preparation, inhibits cortical pain processing by a centrifugal mechanism.

Modulation of laser-evoked potentials by experimental cutaneous tonic pain.

The present study aimed to investigate whether tonic cutaneous pain exerts any effect on the cortical processing of nociceptive input and if this effect may involve only body parts in pain. Tonic cutaneous pain was obtained in nine healthy human subjects by infusion of a hypertonic saline (5%) in the s.c. tissue over the hypothenar muscles (10 ml/h for 20 min). Nociceptive cutaneous CO2 laser-evoked potentials were recorded after stimulation of the right hand dorsum, which was adjacent to the painful area, and the right perioral region, corresponding to the adjacent cortical sensory area. Laser-evoked potentials were obtained before saline injection, at the peak pain and 20 min after pain disappeared. During saline infusion, the laser-evoked pain to right hand stimulation was reduced and the vertex laser-evoked potentials (N2a-P2, mean latency 181 ms and 319 ms for the N2a and the P2 potentials, respectively), which are generated in the anterior cingulate cortex, were significantly decreased in amplitude compared with the baseline. Moreover, the topography of these potentials was modified by cutaneous pain, shifting from the central toward the parietal region. Dipolar modeling showed that the dipolar source in the anterior cingulate cortex moved backward during saline infusion. This result suggests that cutaneous pain may modify the relative activities of the anterior and posterior anterior cingulate cortex parts, which are thought to be devoted to encode different aspects of pain sensation. No laser-evoked potential change was observed after stimulation of the right perioral region, suggesting that functional changes in the nociceptive system are selective for the painful regions and not for areas with cortical proximity.

Evaluation of two different extraction methods for chromatographic determination of bioactive amines in tomato products.

Bioactive amines are organic bases originating from corresponding amino acid which have undergone decarboxylation by putrefactive bacteria or lactic acid bacteria. When formed by microbial enzymatic decarboxylation of amino acids, they are called " biogenic" and can produce detrimental effects on human health. Many techniques have been developed for extraction and/or clean up of bioactive amines in food, including acidic or organic extraction as well as solid phase extraction. This study deals with the comparison of two different extraction methods, homogenizing and matrix solid phase dispersion, for the chromatographic determination of eight non-volatile bioactive amines in tomato-based products (mashed tomato, biological mashed tomato, concentrated tomato pasta and ketchup) very popular in Italian alimentary habits. In both cases, perchloric acid has been used for analytes extraction and the influence of different parameters affecting amine recoveries have been evaluated. After a derivatization step with dansyl-chloride, samples were analyzed for amines quantitative determination using 1,7-diaminoheptane as internal standard on a C(18)-RP-HPLC-UV system. Method performances were tested and good results of linearity, repeatability and recovery were obtained for all the considered amines. The collected data have shown that ketchup contains the highest levels of amines followed by concentrated tomato pasta, biological mashed tomato and common mashed tomato. Moreover, it has been found that in all samples, putrescine is the most abundant amine followed by tyramine, spermidine and tryptamine.

Segmental inhibition of cutaneous heat sensation and of laser-evoked potentials by experimental muscle pain.

The aim of the study was to evaluate the effect of tonic muscle pain evoked by injection of 5% hypertonic saline in the right brachioradialis muscle on the somatosensory sensation of laser-evoked heat pain and laser-evoked potentials. The heat pain pathways were studied in 9 healthy human subjects by recording the scalp potentials evoked by CO(2) laser stimuli delivered on four sites: the skin above the right brachioradialis muscle (ipsilateral local pain), the wrist area where muscle pain was referred in all subjects (ipsilateral referred pain), and two areas on the left arm symmetrical to both local and referred pain (contralateral local pain and contralateral referred pain). Laser-evoked potentials were obtained from 31 scalp electrodes before saline injection, during saline infusion (bolus injection with 0.3 ml saline infused over 20 s, followed by a steady infusion rate of 30 ml/h for the next 25 min), and 20 min after muscle pain had disappeared. While the early N1/P1 component (around 130 ms and 145 ms of latency after stimulation of the skin over the brachioradialis muscle and the wrist, respectively) was not affected by muscle pain, the amplitudes of the later vertex laser-evoked potentials (N2 latency of around 175 ms and 210 ms after stimulation of the skin over the brachioradialis muscle and the wrist, respectively; P2 latency of around 305 ms and 335 ms after stimulation of the skin over the brachioradialis muscle and the wrist, respectively) evoked from ipsilateral local pain, ipsilateral referred pain, and contralateral local pain sites were significantly decreased during muscle pain compared with the baseline recording, while they recovered after pain had disappeared. At the same stimulation sites, the rating of the laser-evoked pain sensation was reduced significantly during muscle pain as compared with the baseline and it recovered after pain had disappeared. On the contrary, muscle pain did not show any effect on both laser-evoked pain and laser-evoked potential amplitude when the contralateral referred pain site was stimulated. The muscle pain inhibitory effect on both heat pain sensation and laser-evoked potential amplitude is probably mediated by an ipsilateral and contralateral segmental mechanism which acts also on the referred pain area, while more general inhibitory mechanisms, such as a distraction effect or a diffuse noxious inhibitory control, are excluded by the absence of any effect of muscle pain on laser-evoked pain and laser-evoked potentials obtained from a remote site, such as the contralateral referred pain area. Since muscle pain induced by hypertonic saline injection is very similar to clinical pain, our results can be useful in understanding the pathophysiology of the somatosensory modifications which can be observed in patients with musculoskeletal pain syndromes.

Tensor veli palatini electromyography with surface electrode applied transnasally.

A new technique is proposed for paratubal electromyography, using a surface, non-invasive, electrode applied transnasally under nasopharyngoscope guidance. This electrode records activity of the tensor veli palatini muscle during swallowing. This technique is of interest for two reasons: endoscopic guidance offers the possibility to check correct positioning of the electrode recording at tensor veli palatini muscle level. Introduction of the non-invasive surface electrode is simple and not painful.

Parietal generators of low- and high-frequency MN (median nerve) SEPs: data from intracortical human recordings.

OBJECTIVE: To identify low and high-frequency median nerve (MN) somatosensory evoked potential (SEP) generators by means of chronically implanted electrodes in the parietal lobe (SI and neighbouring areas) of two epileptic patients. METHODS: Wide-pass short-latency and long-latency SEPs to electrical MN stimulation were recorded in two epileptic patients by stereotactically chronically implanted electrodes in the parietal lobe (SI and neighbouring areas). To study high-frequency responses (HFOs) an off-line digital filtering of depth short-latency SEPs was performed (500-800 Hz, 24 dB roll-off). Spectral analysis was performed by fast Fourier transform. RESULTS: In both patients we recorded a N20/P30 potential followed by a biphasic N50/P70 response. A little negative response in the 100 ms latency range was the last detectable wide-pass SEP in both patients. Two HFOs components (called iP1 and iP2) were detected by mere visual analysis and spectral analysis, and were supposed to be originated within the parietal cortex. CONCLUSIONS: This was the very first study that recorded wide bandpass and high frequency SEPs by electrodes, exploring both the lateral and the mesial part of the parietal lobe and particularly that of the post-central gyrus.

Reduced habituation to experimental pain in migraine patients: a CO(2) laser evoked potential study.

The habituation to sensory stimuli of different modalities is reduced in migraine patients. However, the habituation to pain has never been evaluated. Our aim was to assess the nociceptive pathway function and the habituation to experimental pain in patients with migraine. Scalp potentials were evoked by CO(2) laser stimulation (laser evoked potentials, LEPs) of the hand and facial skin in 24 patients with migraine without aura (MO), 19 patients with chronic tension-type headache (CTTH), and 28 control subjects (CS). The habituation was studied by measuring the changes of LEP amplitudes across three consecutive repetitions of 30 trials each (the repetitions lasted 5 min and were separated by 5-min intervals). The slope of the regression line between LEP amplitude and number of repetitions was taken as an index of habituation. The LEPs consisted of middle-latency, low-amplitude responses (N1, contralateral temporal region, and P1, frontal region) followed by a late, high-amplitude, negative-positive complex (N2/P2, vertex). The latency and amplitude of these responses were similar in both patients and controls. While CS and CTTH patients showed a significant habituation of the N2/P2 response, in MO patients this LEP component did not develop any habituation at all after face stimulation and showed a significantly lower habituation than in CS after hand stimulation. The habituation index of the vertex N2/P2 complex exceeded the normal limits in 13 out of the 24 MO patients and in none of the 19 CTTH patients (P<0.0001; Fisher's exact test). Moreover, while the N1-P1 amplitude showed a significant habituation in CS after hand stimulation, it did not change across repetitions in MO patients. In conclusion, no functional impairment of the nociceptive pathways, including the trigeminal pathways, was found in either MO or CTTH patients. But patients with migraine had a reduced habituation, which probably reflects an abnormal excitability of the cortical areas involved in pain processing.

Influence of cholinergic circuitries in generation of high-frequency somatosensory evoked potentials.

OBJECTIVE: High-frequency oscillations (HFOs) evoked by upper limb stimulation reflect highly synchronised spikes generated in the somatosensory human system. Since acetylcholine produces differential modulation in subgroups of neurons, we would determine whether cholinergic drive influences HFOs. METHODS: We recorded somatosensory evoked potentials (SEPs) from 31 scalp electrodes in 7 healthy volunteers, before and after single administration of rivastigmine, an inhibitor of central acetylcholinesterase. Right median nerve SEPs have been analysed after digital narrow bandpass filtering (500-700 Hz). Raw data were further submitted to Brain Electrical Source analysis (BESA) to evaluate the respective contribution of lemniscal, thalamic and cortical sources. Lastly, we analysed by Fast Fourier transform spectral changes after drug administration in the 10-30 ms latency range. RESULTS: Rivastigmine administration caused a significant increase of HFOs in the 18-28 ms latency range. Wavelets occurring before the onset latency of the conventional N20 SEP did not show any significant change. A similar increase concerned the strength of cortical dipolar sources in our BESA model. Lastly, we found a significant power increase of the frequency peak at about 600 Hz in P3-F3 traces after drug intake. CONCLUSIONS: Our findings demonstrate that the cortical component of HFOs is significantly enhanced by cholinergic activation. Pyramidal chattering cells, which are capable to discharge high-frequency bursts, are mainly modulated by cholinergic inputs; by contrast, acetylcholine does not modify the firing rate of fast-spiking GABAergic interneurons. We thus discuss the hypothesis that cortical HFOs are mainly generated by specialised pyramidal cells.

The human supplementary motor area-proper does not receive direct somatosensory inputs from the periphery: data from stereotactic depth somatosensory evoked potential recordings.

Somatosensory evoked potentials to median nerve (MN) stimulation were recorded by stereotactically implanted electrodes in the frontal lobe of two epileptic patients in order to evaluate whether short-latency cortical responses could be generated in the supplementary motor area (SMA)-proper. In both patients two potentials, called P20 and N30 responses, showed a decreasing amplitude from the most superficial to the deepest contacts and were recorded in the white as well in the grey matter of the frontal lobe. Furthermore, no phase reversal of both P20 and N30 potentials was identifiable along electrode trajectories. Our results suggest that short-latency somatosensory evoked responses recorded in the SMA-proper contralateral to MN stimulation are volume conducted from remote cortical generators.

Cervical cord dysfunction during neck flexion in Hirayama's disease.

Neck flexion may play a role in the pathogenesis of Hirayama disease. Upper limb somatosensory evoked potentials were recorded in five patients with Hirayama disease, six patients with ALS, and 14 healthy subjects. Neck flexion caused a significant amplitude decrease of the N13 cervical response only in patients with Hirayama disease. Direct cord compression or microvascular changes can in theory account for this position-related dysfunction.

Contribution of GABAergic cortical circuitry in shaping somatosensory evoked scalp responses: specific changes after single-dose administration of tiagabine.

OBJECTIVES: To determine whether conventional as well as high-frequency somatosensory evoked potentials (SEPs) to upper limb stimulation are influenced by GABAergic intracortical circuitry. METHODS: We recorded SEPs from 6 healthy volunteers before and after a single-oral administration of tiagabine. Conventional low-frequency SEPs have been obtained after stimulation of the median nerve, as well as after stimulation of the first phalanx of the thumb, which selectively involves cutaneous finger inputs. Median nerve SEPs have been further analyzed after digital narrow-bandpass filtering, to selectively examine high-frequency responses. Lastly, in order to explain scalp SEP distribution before and after tiagabine administration, we performed the brain electrical source analysis (BESA) of raw data. RESULTS: After tiagabine administration, conventional scalp SEPs showed a significant amplitude increase of parietal P24, frontal N24 and central P22 components. Similarly, BESA showed a significant strength increase of the second peak of activation of the first two perirolandic dipoles, which are likely to correspond to the N24/P24 and P22 generators. By contrast, no significant changes of high-frequency SEPs were induced by drug intake. CONCLUSIONS: Our findings support the view that both N24/P24 and P22 SEP components are probably generated by deep spiny cell hyperpolarization, which is strongly increased by inhibitory inputs from GABAergic interneurons. By considering the clear influence of inhibitory circuitry in shaping these SEP components, conventional scalp SEP recording could be useful in the functional assessment of the somatosensory cortex in different physiological and pathological conditions. By contrast, intrinsic firing properties of the cell population generating high-frequency SEP responses are unaffected by the increase of recurrent GABAergic inhibition.

Source generators of the early somatosensory evoked potentials to tibial nerve stimulation: an intracerebral and scalp recording study.

OBJECTIVE: To investigate the location of the cerebral generators of the early scalp somatosensory evoked potentials (SEPs) after tibial nerve stimulation. METHODS: Tibial nerve SEPs were recorded in 15 patients, suffering from Parkinson's disease, who underwent implantation of intracerebral (IC) electrodes in the subthalamic nucleus, in the globus pallidum or in the thalamic ventralis intermediate nucleus. SEPs were recorded both from the scalp surface and from the IC leads. RESULTS: The lemniscal P30 response was recorded by all the electrodes. The IC waveforms included a negative N40IC response, followed by a positive (P50IC) and a negative (N60IC) potential. The N40IC, the P50IC and the N60IC potentials did not differ in latency from the P40, the N50 and the P60 responses recorded by the Cz electrode. In 6 patients, in which SEPs were recorded also during the voluntary movement of the stimulated foot (active gating), an amplitude reduction of the SEP components following the P30 potential was observed during movement at the vertex and in the IC traces. Instead, in the contralateral temporal traces the SEP components (N40temp and P50temp) were not modified by active gating, and in the ipsilateral parietal traces only the positive potentials at about 60ms of latency was decreased. CONCLUSIONS: Two differently oriented generators are active in the contralateral hemisphere at both 40 and 50ms of latency after tibial nerve stimulation. One source is oriented perpendicularly to the mesial hemispheric surface and generates the potentials recorded by the contralateral temporal and the ipsilateral parietal leads; the other dipolar source is radial to the hemispheric convexity, and generates the potentials at the vertex and those recorded by the IC electrodes.

Inhibition of biceps brachii muscle motor area by painful heat stimulation of the skin.

ract This study aimed to evaluate whether painful cutaneous stimuli can affect specifically the excitability of the arm proximal muscle motor area. The motor evoked potentials (MEPs), recorded from the right biceps brachii muscle after either transcranial magnetic or electrical anodal stimulation of the left primary motor (MI) cortex, were conditioned by painful CO2 laser stimuli delivered either on the right hand dorsum or on the lateral surface of the right arm. Painful CO2 laser stimuli delivered on the hand skin reduced significantly the amplitude of MEPs evoked by the transcranial magnetic stimulation of the contralateral MI area, while the MEP amplitude was not significantly modified by CO2 laser pulses delivered on the arm skin. The inhibitory effect followed the arrival of the nociceptive inputs to the cerebral cortex. The amplitude of MEPs evoked by anodal electrical stimulation of the motor cortex was not decreased by conditioning painful stimuli delivered on the hand dorsum. Since the magnetic stimulation led to transynaptic activation of pyramidal neurons, while the anodal stimulation activated directly corticospinal axons, our findings suggest that CO2 laser pulses delivered on the hand are able to inhibit the arm proximal muscle motor area.

Functional changes of the primary somatosensory cortex in patients with unilateral cerebellar lesions.

Although cerebellar lesions do not cause evident sensory deficits, it has been suggested recently that the cerebellum might play a role in sensory acquisition and discrimination. To determine whether the cerebellum influences the early phases of cortical somatosensory processing, we recorded cortical somatosensory evoked potentials after median nerve stimulation in five patients with unilateral cerebellar damage. We also performed a dipolar source analysis of traces by means of brain electrical source analysis. In all patients, the amplitude of the frontal N24 and parietal P24 components, as well as the strength of the corresponding dipolar sources, were significantly smaller after stimulation of the symptomatic side. These neurophysiological findings indicate that the primary somatosensory cortical processing is altered after contralateral cerebellar damage. They represent the first indication of a possible substrate for the reduction in cerebral blood flow observed in the parietal cortex after cerebellar lesion. Furthermore, the present data allow characterization of the functional influence of the cerebellar input to the primary somatosensory cortex as specifically acting over the inhibitory components of somatosensory processing.