Solanum lycopersicum (Tomato)-Derived Nanovesicles Accelerate Wound Healing by Eliciting the Migration of Keratinocytes and Fibroblasts.

Plant-derived nanovesicles have been considered interesting in medicine for their breakthrough biological effects, including those relevant to wound healing. However, tomato-derived nanovesicles (TDNVs) have not been studied for their effects on wound closure yet. TDNVs were isolated from Solanum lycopersicum (var. Piccadilly) ripe tomatoes by ultracentrifugation. Extract (collected during the isolation procedure) and NVs (pellet) were characterized by transmission electron microscopy and laser Doppler electrophoresis. Wound healing in the presence of Extract or NVs was analyzed by a scratch assay with monocultures of human keratinocytes (HUKE) or NIH-3T3 mouse fibroblasts. Cell proliferation and migration were studied by MTT and agarose spot assay, respectively. The vesicles in the Extract and NV samples were nanosized with a similar mean diameter of 115 nm and 130 nm, respectively. Both Extract and NVs had already accelerated wound closure of injured HUKE and NIH-3T3 monocultures by 6 h post-injury. Although neither sample exerted a cytotoxic effect on HUKE and NIH-3T3 fibroblasts, they did not augment cell proliferation. NVs and the Extract increased cell migration of both cell types. NVs from tomatoes may accelerate wound healing by increasing keratinocyte and fibroblast migration. These results indicate the potential therapeutic usefulness of TDNVs in the treatment of chronic or hard-to-heal ulcers.

A Vaccine against Cancer: Can There Be a Possible Strategy to Face the Challenge? Possible Targets and Paradoxical Effects.

Is it possible to have an available vaccine that eradicates cancer? Starting from this question, this article tries to verify the state of the art, proposing a different approach to the issue. The variety of cancers and different and often unknown causes of cancer impede, except in some cited cases, the creation of a classical vaccine directed at the causative agent. The efforts of the scientific community are oriented toward stimulating the immune systems of patients, thereby preventing immune evasion, and heightening chemotherapeutic agents effects against cancer. However, the results are not decisive, because without any warning signs, metastasis often occurs. The purpose of this paper is to elaborate on a vaccine that must be administered to a patient in order to prevent metastasis; metastasis is an event that leads to death, and thus, preventing it could transform cancer into a chronic disease. We underline the fact that the field has not been studied in depth, and that the complexity of metastatic processes should not be underestimated. Then, with the aim of identifying the target of a cancer vaccine, we draw attention to the presence of the paradoxical actions of different mechanisms, pathways, molecules, and immune and non-immune cells characteristic of the tumor microenvironment at the primary site and pre-metastatic niche in order to exclude possible vaccine candidates that have opposite effects/behaviors; after a meticulous evaluation, we propose possible targets to develop a metastasis-targeting vaccine. We conclude that a change in the current concept of a cancer vaccine is needed, and the efforts of the scientific community should be redirected toward a metastasis-targeting vaccine, with the increasing hope of eradicating cancer.

Assessment of Stress Salivary Markers, Perceived Stress, and Shift Work in a Cohort of Fishermen: A Preliminary Work.

Due to work-related stress, today, work itself represents a daily challenge that must be faced in many occupations. While, in the past, the scientific community has focused on the helping professions, since, an increasing number of professions have since been investigated. Therefore, different approaches exist in order to assess this disorder, representing a scientific field wherein biological and psychological dimensions both need to be evaluated. In this paper, we consider three biological salivary markers: interleukin 1 beta (IL-1ß), cortisol, and melatonin. The choice derives from recent contributions to the literature in which the interplay between these markers has been verified. Briefly, such interplay could explain how the central nervous, endocrine, and immune systems communicate with each other, supporting a holistic concept of person. In 30 marine fishermen from the Apulia region of Italy, perceived stress was measured using the Professional Stress Scale (PSS) and sleep disturbances were assessed through the Pittsburgh Sleep Quality Index (PSQI). Salivary markers were collected at 8:00 a.m. and 2:00 p.m. Those subjects reporting sleep disturbance and having altered scores in two PSS subclasses, home-work conflict and self-esteem, presented inverted salivary melatonin and cortisol nictemeral rhythms (with regard to melatonin levels at 8:00 a.m., those workers reporting values higher than the median showed 64.1% versus 48.6% home-work conflict with respect to cortisol levels, subjects having an inverted circadian rhythm showed 69.9% versus 52.5% home-work conflict, and these values resulted 47.7% versus 25.3% when the self-esteem was considered). As regards melatonin, PSQI score is statistically different in the two groups of subjects as identified by median melatonin at 8:00 a.m.; specifically, the subjects who had mean values higher than the median shared higher PSQI scores (10.8 versus 9.8). The same subjects reported more frequent home-work conflict and more sleep disorders. We found a negative correlation between IL-1ß at 8:00 a.m. and Cortdiff (the difference between cortisol at 8:00 a.m.-cortisol at 2:00 p.m.), and that high IL-1ß at 8:00 a.m. was associated with low Cortdiff. Based on our results we would like to propose this approach in health surveillance, in order to prevent mental and/or physical disorders, however our study is surely preliminary. The interesting perspectives and hypotheses cited in this paper, in which the roles of IL-1ß and norepinephrine appear central and important, could remain hypothetical if not supported by more robust observation in order to produce, truly, new knowledge. In the future we will deepen this study with a larger sample, and if these results will be confirmed, this approach could allow preventing, not only mental and physical disorders, but also immuno-mediated diseases, and, perhaps, cancer.

Work Stress, Mental Health and Validation of Professional Stress Scale (PSS) in an Italian-Speaking Teachers Sample.

This study aimed validate the Italian version of the Professional Stress Scale (PSS). A questionnaire was translated into Italian and administered to two sample groups. The first group (n = 200) was the control group and the second (n = 1137) the experimental group. The participants in the study were students enrolled in a special needs training teacher course or a specialization course that aims to train support teachers. The study conducted two analyses; factor and reliability analyses. The factor analysis utilized the Kaiser-Meyer-Olkin (KMO) test which had a result of 0.925 for the scale; this was above the acceptable value of 0.7. The research studied 33 items and the BTS was significant for the 33 items scale (χ2 (528) = 4353.508, p < 0.001). Moreover, five eigenvalues greater than 1 were identified in the data, whereas the total variance explained was 63.7%. The reliability test utilized the Cronbach's Alpha score (0.936) of the scale and the value is calculated based on the response of 1106 individuals. The value is well above the value of 0.80, which indicates a high internal consistency level of the different items of the scale. This study showed that the Italian version of the PSS is a reliable and valid measure that can be used for research and clinical purposes.

How Cells Communicate with Each Other in the Tumor Microenvironment: Suggestions to Design Novel Therapeutic Strategies in Cancer Disease.

Connexin- and pannexin (Panx)-formed hemichannels (HCs) and gap junctions (GJs) operate an interaction with the extracellular matrix and GJ intercellular communication (GJIC), and on account of this they are involved in cancer onset and progression towards invasiveness and metastatization. When we deal with cancer, it is not correct to omit the immune system, as well as neglecting its role in resisting or succumbing to formation and progression of incipient neoplasia until the formation of micrometastasis, nevertheless what really occurs in the tumor microenvironment (TME), which are the main players and which are the tumor or body allies, is still unclear. The goal of this article is to discuss how the pivotal players act, which can enhance or contrast cancer progression during two important process: "Activating Invasion and Metastasis" and the "Avoiding Immune Destruction", with a particular emphasis on the interplay among GJIC, Panx-HCs, and the purinergic system in the TME without disregarding the inflammasome and cytokines thereof derived. In particular, the complex and contrasting roles of Panx1/P2X7R signalosome in tumor facilitation and/or inhibition is discussed in regard to the early/late phases of the carcinogenesis. Finally, considering this complex interplay in the TME between cancer cells, stromal cells, immune cells, and focusing on their means of communication, we should be capable of revealing harmful messages that help the cancer growth and transform them in body allies, thus designing novel therapeutic strategies to fight cancer in a personalized manner.

Molecular links between endocrine, nervous and immune system during chronic stress.

INTRODUCTION: The stress response is different in various individuals, however, the mechanisms that could explain these distinct effects are not well known and the molecular correlates have been considered one at the time. Particular harmful conditions occur if the subject, instead to cope the stressful events, succumb to them, in this case, a cascade reaction happens that through different signaling causes a specific reaction named "sickness behaviour." The aim of this article is to review the complex relations among important molecules belonging to Central nervous system (CNS), immune system (IS), and endocrine system (ES) during the chronic stress response. METHODS: After having verified the state of art concerning the function of cortisol, norepinephrine (NE), interleukin (IL)-1ß and melatonin, we describe as they work together. RESULTS: We propose a speculative hypothesis concerning the complex interplay of these signaling molecules during chronic stress, highlighting the role of IL-1ß as main biomarker of this effects, indeed, during chronic stress its increment transforms this inflammatory signal into a nervous signal (NE), in turn, this uses the ES (melatonin and cortisol) to counterbalance again IL-1ß. During cortisol resistance, a vicious loop occurs that increments all mediators, unbalancing IS, ES, and CNS networks. This IL-1ß increase would occur above all when the individual succumbs to stressful events, showing the Sickness Behaviour Symptoms. IL-1ß might, through melatonin and vice versa, determine sleep disorders too. CONCLUSION: The molecular links here outlined could explain how stress plays a role in etiopathogenesis of several diseases through this complex interplay.

Gap Junction Intercellular Communication in the Carcinogenesis Hallmarks: Is This a Phenomenon or Epiphenomenon?

If occupational tumors are excluded, cancer causes are largely unknown. Therefore, it appeared useful to work out a theory explaining the complexity of this disease. More than fifty years ago the first demonstration that cells communicate with each other by exchanging ions or small molecules through the participation of connexins (Cxs) forming Gap Junctions (GJs) occurred. Then the involvement of GJ Intercellular Communication (GJIC) in numerous physiological cellular functions, especially in proliferation control, was proven and accounts for the growing attention elicited in the field of carcinogenesis. The aim of the present paper is to verify and discuss the role of Cxs, GJs, and GJIC in cancer hallmarks, pointing on the different involved mechanisms in the context of the multi-step theory of carcinogenesis. Functional GJIC acts both as a tumor suppressor and as a tumor enhancer in the metastatic stage. On the contrary, lost or non-functional GJs allow the uncontrolled proliferation of stem/progenitor initiated cells. Thus, GJIC plays a key role in many biological phenomena or epiphenomena related to cancer. Depending on this complexity, GJIC can be considered a tumor suppressor in controlling cell proliferation or a cancer ally, with possible preventive or therapeutic implications in both cases.

Gap Junctions Are Involved in the Rescue of CFTR-Dependent Chloride Efflux by Amniotic Mesenchymal Stem Cells in Coculture with Cystic Fibrosis CFBE41o- Cells.

We previously found that human amniotic mesenchymal stem cells (hAMSCs) in coculture with CF immortalised airway epithelial cells (CFBE41o- line, CFBE) on Transwell® filters acquired an epithelial phenotype and led to the expression of a mature and functional CFTR protein. In order to explore the role of gap junction- (GJ-) mediated intercellular communication (GJIC) in this rescue, cocultures (hAMSC : CFBE, 1 : 5 ratio) were studied for the formation of GJIC, before and after silencing connexin 43 (Cx43), a major component of GJs. Functional GJs in cocultures were inhibited when the expression of the Cx43 protein was downregulated. Transfection of cocultures with siRNA against Cx43 resulted in the absence of specific CFTR signal on the apical membrane and reduction in the mature form of CFTR (band C), and in parallel, the CFTR-dependent chloride channel activity was significantly decreased. Cx43 downregulation determined also a decrease in transepithelial resistance and an increase in paracellular permeability as compared with control cocultures, implying that GJIC may regulate CFTR expression and function that in turn modulate airway epithelium tightness. These results indicate that GJIC is involved in the correction of CFTR chloride channel activity upon the acquisition of an epithelial phenotype by hAMSCs in coculture with CF cells.

Possible Mechanisms of Mercury Toxicity and Cancer Promotion: Involvement of Gap Junction Intercellular Communications and Inflammatory Cytokines.

A number of observations indicate that heavy metals are able to alter cellular metabolic pathways through induction of a prooxidative state. Nevertheless, the outcome of heavy metal-mediated effects in the development of human diseases is debated and needs further insights. Cancer is a well-established DNA mutation-linked disease; however, epigenetic events are perhaps more important and harmful than genetic alterations. Unfortunately, we do not have reliable screening methods to assess/validate the epigenetic (promoter) effects of a physical or a chemical agent. We propose a mechanism of action whereby mercury acts as a possible promoter carcinogen. In the present contribution, we resume our previous studies on mercury tested at concentrations comparable with its occurrence as environmental pollutant. It is shown that Hg(II) elicits a prooxidative state in keratinocytes linked to inhibition of gap junction-mediated intercellular communication and proinflammatory cytokine production. These combined effects may on one hand isolate cells from tissue-specific homeostasis promoting their proliferation and on the other hand tamper the immune system defense/surveillance checkmating the whole organism. Since Hg(II) is not a mutagenic/genotoxic compound directly affecting gene expression, in a broader sense, mercury might be an example of an epigenetic tumor promoter or, further expanding this concept, a "metagenetic" effector.

Stress-related temporary hearing loss--evaluation of bio-humoral parameters: forensic and criminological applications.

The body-alarm reaction results from the activation of hypothalamic-pituitary-adrenal axis, which can lead to physio-psychological phenomena such as an exclusion/occlusion of the sense of hearing. One hypothesis to explain this alteration consists in a hydromechanical dysfunction of the internal ear attributable to antidiuretic hormone. In this study, we evaluated the perception of acoustic stimuli administered in stressful conditions in 14 phobic patients and in 20 healthy subjects, in order to assess the influence of stress on perceiving capabilities. We also measured the concentration of salivary cortisol and IL-1ß and neurovegetative parameters to objectivise and quantify the physiological reactions. Our results show a worse perception of the frequencies of the human voice under stress; these findings could have a dual value: in the legal field, concerning criminal liability, and on the operative context, regarding the efficiency of verbal communication among law enforcement officers in situations inducing intense emotional stress.

Subcytotoxic mercury chloride inhibits gap junction intercellular communication by a redox- and phosphorylation-mediated mechanism.

Gap junctions play a central role in coordinating intercellular signal-transduction pathways to control tissue homeostasis. Deregulation of gap junctional intercellular communication is a common phenotype of cancer cells and supports its involvement in the carcinogenesis process. Many carcinogens, like environmental heavy-metal chemical pollutants, are known to activate various signal transduction mechanisms and modulate GJIC. They act as tumor promoters on preexisting "initiated" cells, rather than as genotoxic initiators, albeit their mode of action is often unknown. In this study we investigated the effect of Hg(II) (HgCl(2)) on GJIC in cultured human keratinocytes. It is shown that subcytotoxic concentrations of HgCl(2) as low as 10 nM cause inhibition of the GJIC, assessed by dye transfer assay, despite enhanced expression of connexins. In addition, HgCl(2)-treated keratinocytes exhibited a decrease of free thiols and accumulation of mitochondria-derived reactive oxygen species, albeit no effect on the respiratory chain activity was observed. Treatment of HgCl(2)-exposed keratinocytes with the PKC inhibitor calphostin C and with all-trans retinoic acid resulted in rescue of the mitochondrial ROS overproduction and full recovery of the GJIC. Similar results were obtained with the PKA activator db-cAMP. Overall, the presented results support a cross-talk between the altered intracellular redox tone and PKA- and PKC-mediated signaling in HgCl(2)-challenged keratinocytes. These events, although not cytotoxic, lead to inhibition of GJIC and possibly to carcinogenic priming.

Supercritical CO(2)-extracted tomato Oleoresins enhance gap junction intercellular communications and recover from mercury chloride inhibition in keratinocytes.

A nutritionally relevant phytochemical such as lycopene, found in tomatoes and other fruits, has been proposed to have health-promoting effects by modulating hormonal and immune systems, metabolic pathways, and gap junction intercellular communication (GJIC). This work analyzes lycopene extracts, obtained from tomato and tomato added with grape seeds by using a safe and environmentally friendly extraction process, based on supercritical carbon dioxide technology (S-CO(2)). Analysis of the innovative S-CO(2)-extracted oleoresins showed peculiar chemical composition with high lycopene concentration and the presence of other carotenoids, lipids, and phenol compounds. The oleoresins showed a higher in vitro antioxidant activity compared with pure lycopene and beta-carotene and the remarkable ability to enhance the GJIC and to increase cx43 expression in keratinocytes. The oleoresins, (0.9 microM lycopene), were also able to overcome, completely, the GJIC inhibition induced by 10 nM HgCl(2), mercury(II) chloride, suggesting a possible action mechanism.

Salivary endothelin and vascular disorders in vibration-exposed workers.

OBJECTIVES: This study investigated the relation between salivary endothelin, vibration exposure, and vascular disorders in a group of forestry workers. METHODS: Altogether 54 forestry workers and 52 controls underwent a medical examination and a cold test with measurement of the percentage of change in finger systolic blood pressure after finger cooling from 30 degrees C to 10 degrees C (FSBP% (10 degrees)). Salivary endothelin concentration (ET(1-21), in fmol/ml) was measured by a commercially available enzyme-linked immunosorbent assay before and after the cold challenge. The anamnestic diagnosis of vibration-induced white finger (VWF), assisted by color charts, was based on the Stockholm Workshop criteria. RESULTS: Six forestry workers (11%) and one control (2%) reported white fingers. Before the cold challenge, the salivary ET(1-21) concentration was significantly greater in the VWF workers than in the controls (P=0.036). The cold response of digital arteries was stronger in the VWF workers than in the controls (P<0.001) and the asymptomatic forestry workers (P=0.008). After the cold test, there was a small, not significant, increase in the salivary ET(1-21) concentration in both the controls and the forestry workers. For the latter, the salivary ET(1-21) concentration was significantly associated with both daily and total operating time with vibrating tools. A significant inverse relation between FSBP% (10 degrees )and the salivary ET(1-21) concentration was observed for the forestry workers with an abnormal cold response in their digital arteries. CONCLUSIONS: This study showed an association between salivary ET(1-21) concentration, daily and cumulative vibration exposure, and vascular disorders in the fingers of professional forestry workers. Since ET(1-21) can induce powerful and long-lasting constriction of human vessels, these findings suggest a possible role of this vasopeptide in the pathogenesis of VWF.

Mercury modulates interplay between IL-1beta, TNF-alpha, and gap junctional intercellular communication in keratinocytes: mitigation by lycopene.

Gap junctional intercellular communication (GJIC) is used to control cell proliferation. It is not surprising then that a lack of GJIC (i.e., during loss of contact inhibition among adjacent cells) is associated with cancer promotion/progression. There also seems to be a link between ineffective GJIC and increases in inflammatory events. Interestingly, many cytokines released during an inflammatory response also have critical roles in cancer cell survival. Specifically, TNFalpha and IL-1beta are important for initiating/augmenting CD8(+)- and NK-cell mediated killing; however, in what appears counterintuitive, each--at times--can act to protect cancer cells against apoptosis, a major mechanism for cell killing from within. It is thus plausible to assume that certain toxicants might act as cancer promoters in manners distinct from/augmentive of direct effects on DNA, i.e., by concurrently altering GJIC and cytokine formation in host or microenvironment of a cancer cell. Our research has evaluated effects of many toxicants upon keratinocytes; in particular, we have examined effects of mercury on GJIC and on TNFalpha and IL-1beta levels in (and secretion by) these cells. In the studies here, a tomato preparation (i.e., an oleoresin) bearing the antioxidant carotenoid lycopene was examined for its effects on GJIC and cytokine formation by keratinocytes in general, and its potential ability to mitigate/reverse the toxic effects of mercury in the cells in particular. It was shown that a 4-hr treatment with the oleoresin (containing 56, 6 nM lycopene) re-established GJIC among--and increased the formation of IL-1beta and TNFalpha that had been significantly reduced within--keratinocytes that had been pre-treated for 24 hr with 10 nM HgCl(2). These results show that effects of mercury likely depend on some level of oxidative stress and that its potential effects on keratinocyte GJIC and cytokine concentrations could, in an exposed host, be mitigated/reversed by increased dietary intake of carotenoids like lycopene.

Effect of psychological stress on salivary interleukin-1beta in psoriasis.

BACKGROUND: Psoriasis is a chronic inflammatory skin disease in which several Th1 cytokines such as interleukin-1 beta (IL-beta) have been shown to play a pivotal role. Psychological stress has also been implicated in triggering or exacerbating the disease. METHODS: Salivary IL-1beta and cortisol levels of 25 patients with psoriasis were compared with those of 50 age- and sex-matched healthy controls under basal conditions and after a standardized stressful procedure including mental arithmetics and the Stroop Color-Word Naming Test. RESULTS: At baseline, mean IL-1beta levels were higher in patients with psoriasis than controls (p <0.001), whereas mean cortisol levels did not differ significantly between groups. Although IL-1beta levels increased after stress among controls, they did not increase among patients with psoriasis, with a significant group-by-time interaction (p <0.01). After stress, cortisol levels were significantly increased in both groups as compared with baseline (p < or =0.001), without any group-by-time interaction. Perceived stress was similar among psoriatic patients and controls. There was no significant correlation between changes in IL-1beta and changes in cortisol. CONCLUSIONS: The higher basal IL-1beta levels among psoriatic patients suggest that its production is increased. Changes in proinflammatory cytokine activity in psoriatic skin may play an important role in propagating inflammation. The blunted response of IL-1beta to stress observed in psoriatic patients may reflect a "ceiling effect", or be ascribed to a defective response of the immune system to adrenergic stimuli.

Application of a latent variable model for a multicenter study on early effects due to mercury exposure.

A latent variable model was applied to the results of an Italian multicenter nation-wide cross-sectional study to assess the earliest health effects due to mercury (Hg) exposure caused by occupation, dental amalgams (DENTAM) and fish eating (FISH). The studied population included subjects recruited from four different geographical areas. A total number of 122 workers from chloroalkali plants and production of thermometers and neon lamps formed the occupationally exposed group, whereas 196 subjects, recruited from the same areas and not occupationally exposed to mercury, formed the control group. Neuropsychological functions were assessed with neurobehavioral testing including vigilance, motor and cognitive function, tremor measurements, and with symptoms concerning neuropsychological and mood assessment. Neuroendocrine function was examined with the measurement of prolactin (PRL) level. Parameters of immunological and renal function were also measured. The target population was characterized by the number and surface of dental amalgams and consumption of fish. In the exposed workers the average urinary mercury (U-Hg) was 10.4+/-6.9 (geometric mean 8.3, range 0.2-35.2) microg/g creatinine, whereas in the control group it was 1.9+/-2.8 (geometric mean 1.2, range 0.1-33.2) microg/g creatinine. The preliminary results indicated that finger tapping (FT) and the Branches alternate movement task (BAMT) coordination test were associated with the occupational exposure (OCCEXP). PRL was significantly decreased among the exposed workers, and inversely related to U-Hg. Among the immunological and renal parameters, cytokine serum interleuchin-8 (sIL8) and beta(2) micro globulin (beta(2)MG) were lower in the exposed group and negatively correlated to U-Hg. Small-size fish consumption was associated to a beneficial effect on symptoms reporting. No effects were observed concerning dental amalgams. After first evaluating the relationship between mercury exposure and each indicator of effect, further assessment was performed to identify the earliest effects related to mercury exposure among those who resulted in being associated in the preliminary elaboration. Two latent variables "exposure" and "effect" were identified, integrating respectively the different forms of exposure (occupational, due to dental amalgams and fish consumption) and the indicators of effects (FT, BAMT, PRL, sIL8, beta(2)MG). Confounding factors (age, alcohol, body mass index (BMI)) were considered in the same model. This further analysis showed that an inverse association of occupational exposure to mercury with PRL and BAMT, with Hg-U mediating the effect on PRL, was predominant with respect to the other form of mercury exposure, the other indicators of effect and the confounders. In conclusion, this study supports the finding of alterations of neuroendocrine secretion and motor coordination at very low occupational exposure levels of inorganic mercury, below the current ACGIH Biological Exposure Index. These changes occur at lower levels than other subtle effects on the renal function and the immunitary system. On the contrary, dental amalgams and small-size fish consumption do not seem to be associated to any adverse health effect at these exposure levels.