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White adipose tissue/brown adipose tissue trans-differentiation is one of the main study targets for therapies against obesity and metabolic diseases. In recent years, numerous molecules able to induce such trans-differentiation have been identified; however, their effect in obesity therapies has not been as expected. In the present study, we investigated whether myo-inositol and its stereoisomer D-chiro-inositol could be involved in the browning of white adipose tissue. Our preliminary results clearly indicate that both, at 60 μM concentration, induce the upregulation of uncoupling protein 1 mRNA expression, the main brown adipose tissue marker, and increase mitochondrial copy number as well as oxygen consumption ratio. These changes demonstrate an activation of cell metabolism. Therefore, our results show that human differentiated adipocytes (SGBS and LiSa-2), assume the features typical of brown adipose tissue after both treatments. Furthermore, in the cell lines examined, we proved that D-chiro-inositol and myo-Inositol induce an increase in the expression of estrogen receptor mRNAs, suggesting a possible modulation by these isomers. We also found an increase in the mRNA of peroxisome proliferator-activated receptor gamma, a very important target in lipid metabolism and metabolic diseases. Our results open new opportunities for the use of inositols in therapeutic strategies to counteract obesity and its metabolic complications.

Myo-inositol and D-chiro-inositol are insulin sensitising agents. In the ovary, myo-inositol acts as second messenger of Follicle Stimulating Hormone (FSH). Both molecules were administered to Polycystic Ovary Syndrome (PCOS) women. The gynaecologist Vittorio Unfer was the first to give specific value to myo-inositol for the treatment of PCOS: this important innovation opened new ways of research to identify efficient therapies based on myo-inositol alone or with low doses of D-chiro-inositol. Significant successes were also gained using myo-inositol in treating male and female infertility. Unfer's researches allowed to identify "the D-Chiro-Inositol Paradox in the Ovary" and the best myo-inositol/D-chiro-inositol ratio (40:1) for the treatment of PCOS. Furthermore, his studies allowed to improve the inositol's efficacy using alpha-lactalbumin. As shown in this review, the main stages of Unfer's scientific career have been closely intertwined with important phases of the recent pharmacological research about the topic.

Study question Low doses of D-Chiro-Inositol are beneficial in the treatment of a PCOS mouse model. However, high doses are detrimental for ovarian histology/function. Is D-Chiro-Inositol toxic for the mammalian ovary? Summary answer Five mg/day D-Chiro-Inositol for 21 days produced PCOS-like histological/hormonal features. Ten/20 mg/day for 21 days induced ovarian/hormonal states resembling those typical of aged mice. What is known already Administration of Myo-Inositol and D-Chiro-Inositol combined according to their plasma molar ratio of 40:1 has beneficial effects in the management of human PCOS. We confirmed the efficacy of this formulation, containing 0.2 mg/day D-Chiro-Inositol, in a mouse model of PCOS. However, formulations containing higher amounts of DCIns had negative effects on ovarian histology and mouse fertility. We investigated possible ovarian toxicity of D-Chiro-Inositol, studying its effects after administration to 30-day-old female mice for 21 days. Young adult mice reproduced the condition of young women possibly facing reproductive/metabolic problems, such as PCOS. Study design, size, duration The effects of various doses of D-Chiro-Inositol were analysed on mouse ovarian histology, serum testosterone levels and expression of the ovarian enzyme aromatase. The 21-day period follows normal protocols of pharmacologic PCOS induction in the mouse and spans five ovulatory cycles. Doses employed, 5, 10, 20 mg/day, correspond to doses of 1200, 2400, 4800 mg/day in humans. The first dose is in the range of 1000–1500 mg/day currently prescribed to PCOS patients in clinical practice. Participants/materials, setting, methods Five mice/treatment were provided with water administering various doses of D-Chiro-Inositol or 0,5 mg/day letrozole as PCOS-positive controls, for 21 days. At the end of the period, ovulatory cycles were analysed by observations of vaginal cells after vaginal smears; ovarian histology was evaluated by sectioning, hematoxylin-eosin staining and light-transmission microscopy; serum testosterone levels were measured by ELISA; and expression of the ovarian enzyme aromatase was assayed by Western Blots. Main results and the role of chance The estrus cycle progressed normally in negative control mice, but was arrested at day 8–10 in the majority of mice under all pharmacologic treatments. Uteri of negative control mice displayed the typical aspect of mature and cycling animals. Uteri of all other mice had an immature/metestrus-diestrus-like aspect, typical of non-cycling animals. Ovaries of negative control mice showed a normal presence of primary, secondary and tertiary follicles containing a growing oocyte, and of corpora lutea. Ovaries from mice treated with 5 mg/day D-Chiro-Inositol or 0,5 mg/day letrozole had apparently normal primary and secondary follicles but also cystic tertiary follicles resembling those found in human PCOS. Ovaries from mice treated with 10 or 20 mg/day D-Chiro-Inositol had scarce primary and secondary follicles, a very limited number of tertiary follicles and no cystic follicles, but large follicles/areas with diffused cell proliferation. The typical ovarian structure was lost, especially in the highest dosage. Treatments with 5 mg/day D-Chiro-Inositol and 0,5 mg/day letrozole increased serum testosterone levels above those of negative control mice, but the former reduced, while the latter increased aromatase levels relative to negative controls. Other treatments had no apparent effects on either testosterone or aromatase levels. Our experimental paradigm makes the role of chance highly improbable. Limitations, reasons for caution The strength of our study relies on the use of an animal model representative of general human tissue organisation and physiological pathways. One weakness consists in the lack of data on serum estrogen levels, due to the paucity of blood provided by a single mouse and the ELISA sensitivity. Wider implications of the findings: Under all experimental conditions, D-Chiro-Inositol negatively affected ovarian histology and function. Notwithstanding physiological/biochemical differences between mice and humans, caution is therefore recommended when administering D-Chiro-Inositol to PCOS patients at doses corresponding to those we employed in the mouse and/or for long periods, since it may result ineffective or even toxic. Trial registration number Not applicable

Myo-inositol and D-chiro-inositol are insulin sensitising agents. In the ovary, myo-inositol acts as second messenger of Follicle Stimulating Hormone (FSH). Both molecules were administered to Polycystic Ovary Syndrome (PCOS) women. The gynaecologist Vittorio Unfer was the first to give specific value to myo-inositol for the treatment of PCOS: this important innovation opened new ways of research to identify efficient therapies based on myo-inositol alone or with low doses of D-chiro-inositol. Significant successes were also gained using myo-inositol in treating male and female infertility. Unfer's researches allowed to identify "the D-Chiro-Inositol Paradox in the Ovary" and the best myo-inositol/D-chiro-inositol ratio (40:1) for the treatment of PCOS. Furthermore, his studies allowed to improve the inositol's efficacy using alpha-lactalbumin. As shown in this review, the main stages of Unfer's scientific career have been closely intertwined with important phases of the recent pharmacological research about the topic.

Administration of 1000–1500 mg/day D-Chiro-Inositol (DCIns) or a combination of Myo-Inositol (MyoIns) and DCIns in their plasma molar ratio (40:1) for three or more months are among recommended treatments for metabolic syndrome and/or Polycystic Ovary Syndrome (PCOS). We previously confirmed the efficacy of this formulation (8.2 mg/day MyoIns and 0.2 mg/day DCIns for 10 days) in a mouse PCOS model, but also observed negative effects on ovarian histology and function of formulations containing 0.4–1.6 mg/day DCIns. We therefore analyzed effects of higher doses of DCIns, 5, 10 and 20 mg/day, administered to young adult female mice for 21 days, on ovarian histology, serum testosterone levels and expression of the ovarian enzyme aromatase. Five mg/day DCIns (human correspondence: 1200 mg/day) altered ovarian histology, increased serum testosterone levels and reduced the amount of aromatase of negative controls, suggesting the induction of an androgenic PCOS model. In contrast, 10–20 mg/day DCIns (human correspondence: 2400–4800 mg/day) produced ovarian lesions resembling those typical of aged mice, and reduced serum testosterone levels without affecting aromatase amounts, suggesting a failure in steroidogenic gonadal activity. Notwithstanding physiological/biochemical differences between mice and humans, the observed pictures of toxicity for ovarian histology and function recommend caution when administering DCIns to PCOS patients at high doses and/or for periods spanning several ovulatory cycles.

International audience; Myo-inositol (myo-Ins) and D-chiro-inositol (D-chiro-Ins) are natural compounds involved in many biological pathways. Since the discovery of their involvement in endocrine signal transduction, myo-Ins and D-chiro-Ins supplementation has contributed to clinical approaches in ameliorating many gynecological and endocrinological diseases. Currently both myo-Ins and D-chiro-Ins are well-tolerated, effective alternative candidates to the classical insulin sensitizers, and are useful treatments in preventing and treating metabolic and reproductive disorders such as polycystic ovary syndrome (PCOS), gestational diabetes mellitus (GDM), and male fertility disturbances, like sperm abnormalities. Moreover, besides metabolic activity, myo-Ins and D-chiro-Ins deeply influence steroidogenesis, regulating the pools of androgens and estrogens, likely in opposite ways. Given the complexity of inositol-related mechanisms of action, many of their beneficial effects are still under scrutiny. Therefore, continuing research aims to discover new emerging roles and mechanisms that can allow clinicians to tailor inositol therapy and to use it in other medical areas, hitherto unexplored. The present paper outlines the established evidence on inositols and updates on recent research, namely concerning D-chiro-Ins involvement into steroidogenesis. In particular, D-chiro-Ins mediates insulin-induced testosterone biosynthesis from ovarian thecal cells and directly affects synthesis of estrogens by modulating the expression of the aromatase enzyme. Ovaries, as well as other organs and tissues, are characterized by a specific ratio of myo-Ins to D-chiro-Ins, which ensures their healthy state and proper functionality. Altered inositol ratios may account for pathological conditions, causing an imbalance in sex hormones. Such situations usually occur in association with medical conditions, such as PCOS, or as a consequence of some pharmacological treatments. Based on the physiological role of inositols and the pathological implications of altered myo-Ins to D-chiro-Ins ratios, inositol therapy may be designed with two different aims: (1) restoring the inositol physiological ratio; (2) altering the ratio in a controlled way to achieve specific effects.

The antioxidants role in cell response regulation attracted great interest in the last decades and it is undergoing to a profound reconsideration. The mere concept of "biological antioxidant" has been frequently misconceived or misused, possibly leading to the misinterpretation of some experimental observation. Organosulfur compounds in general and α-lipoic acid, a dithiol molecule, can be considered a typical example of the kind. Reduced α-lipoic acid, dehydrolipoic acid has been in fact originally considered a bona fide, reducing, electron donor molecule. A more recent approach, according to stoichiometric and thermodynamic evidences, lead to a reinterpretation of the biochemical role of "antioxidants". The electrophilic nature of oxidized nucleophilic molecules, including α-lipoic acid, renders more plausible a mechanism based on the ability to activate Nrf2/EpRE mediated hormetic response. In this study, we demonstrate that nmolar concentrations of oxidized α-lipoic acid, but not dehydrolipoic acid, protect human umbilical primary endothelial cells (HUVEC) from TNF-α induced dysfunction, inhibit NF-κB activation and block apoptosis following the activation of Nrf2 transcription factor. Our observations corroborate the concept that the major, if not the unique, mechanism by which α-lipoic acid can non-enzymatically exert its reducing activity is related to the electrophilic nature of the oxidized form.

Introduction: This Experts' opinion provides an updated scientific support to gynecologists, obstetricians, endocrinologists, nutritionists, neurologists and general practitioners on the use of Inositols in the therapy of Polycystic Ovary Syndrome (PCOS) and non-insulin dependent (type 2) diabetes mellitus (NIDDM). Areas covered: This paper summarizes the physiology of Myo-Inositol (MI) and D-Chiro-Inositol (DCI), two important molecules present in human organisms, and their therapeutic role, also for treating infertility. Some deep differences between the physiological functions of MI and DCI, as well as their safety and intestinal absorption are discussed. Updates include new evidence on the efficacy exerted in PCOS by the 40:1 MI/DCI ratio, and the innovative approach based on alpha-lactalbumin to overcome the decreased therapeutic efficacy of Inositols in some patients. Expert opinion: The evidence suggests that MI, alone or with DCI in the 40:1 ratio, offers a promising treatment for PCOS and NIDDM. However, additional studies need to evaluate some still unresolved issues, such as the best MI/DCI ratio for treating NIDDM, the potential cost-effectiveness of reduced gonadotropins administration in IVF due to MI treatment, or the benefit of MI supplementation in ovulation induction with clomiphene citrate in PCOS patients.

It is well known that myo-inositol (MI) and D-chiro-inositol (DCI) are insulin-sensitizing agents, and MI is of proven utility in polycystic ovary syndrome (PCOS). In addition, MI plays a pivotal role in the physiology of reproduction, and has beneficial effects on the development of oocytes, spermatozoa, and embryos. By contrast, DCI has little effect on spermatozoa, but high concentrations in the ovary can negatively affect the quality of oocytes and the blastocyst. Overall, the evidence in the literature supports the beneficial effects of MI in both female and male reproduction, warranting clinical use of MI in assisted reproductive treatment (ART).

Alpha lipoic acid (ALA) is an essential mitochondrial co-factor and, as a free molecule, it can exert multi-level immunomodulatory functions. Both ALA and its reduced form, dihydrolipoic acid (DHLA), are believed to be able to chelate heavy metals, to regenerate essential antioxidants and to repair important molecules damaged by oxidation. The largest part of the effects of ALA/DHLA couple can be explained by a specific stimulatory activity on Nrf2-dependent gene transcription and by the inhibition of NF-kB activity. These features have prompted its use as a drug for several diseases. Areas covered: This article surveys the main features of ALA/DHLA and its therapeutic effects. Its complex and differentiated function cannot simply be reduced to anti-inflammatory, antioxidant and detoxifying action. We highlight its capability to finely modulate several physiological pathways when unbalanced. In particular, we focus our attention on pregnancy, in relation to ALA administration by oral route and by a new formulation for vaginal delivery, in patients with threatened miscarriage. Expert opinion: Future efforts should be devoted to explaining carefully ALA/DHLA mechanism of action to reactivate the physiological balance when modified during pregnancy. On the other hand, ALA safety in pregnant women and its pharmacokinetics by vaginal route, have to be studied in depth. Moreover, ALA efficacy has to be confirmed in a much larger sample of patients.

Background: D-chiro-inositol (DCI) and glucose transporter inhibitors may inhibit myo-inositol (MI) transporters, and the aim is to clinically evaluate their effect on MI absorption. Research design and methods: Fasting 18 healthy volunteers received orally 6000 mg MI, 6000 mg MI with 1000 mg DCI, and 6000 mg MI with SelectSIEVE® Apple PCQ and Sorbitol, Maltodextrin and Sucralose (PCQ-SMS), in three different phases with a washout period of 7 days. At each phase, blood samples were collected before administration, and every 60 minutes until 540 minutes after administration. MI plasma levels (μmol/L) were quantified by gas chromatography-mass spectrometry; maximum plasma concentration (Cmax), time to reach it (Tmax), and the area under the time-concentration curve of MI (AUC 0-540) were evaluated. Results: The Cmax of MI alone (Tmax = 180min) was 1.29-fold higher than those of MI with DCI (Tmax = 180min) (p Conclusions: DCI, glucose transporter inhibitors and sugars, such as sorbitol and maltodextrin, seem to inhibit MI absorption, decreasing MI plasma concentration as compared to MI alone.

BACKGROUND Myo-inositol is a natural molecule with important therapeutic applications and an impaired oral absorption may result in a reduced clinical effect. Aim of this study was to determine if the combined oral administration of α-lactalbumin and myo-inositol in healthy subjects, could increase the plasma level of myo-inositol administered alone. In vitro studies on human differentiated intestinal Caco-2 cells were also conducted to identify the mechanisms involved in myo-inositol absorption. OBJECTIVE The in vivo study was conducted on healthy volunteers in two phases. Subjects received a single oral myo-inositol dose. After 7 days washout, the same subjects were administered a single dose of myo-inositol and α-lactalbumin. Cmax, Tmax and AUC for myo-inositol in plasma were calculated from samples collected at different times. Transepithelial myo-inositol passage, with or without addition of digested α-lactalbumin, was measured in vitro in differentiated Caco-2 cells and compared to transepithelial electrical resistance and phenol red passage. RESULTS The bioavailability of myo-inositol was modified by the concomitant administration of α- lactalbumin. Although peak concentration of myo-inositol at 180 min (Tmax) was similar for both treatments, administration of α-lactalbumin with myo-inositol in a single dose, significantly increased the plasma concentrations of myo-inositol compared to when administered alone. In vitro, myo-inositol absorption in Caco-2 cells was improved in the presence of digested α-lactalbumin, and this change was associated with an increase in tight junction permeability. CONCLUSION Better myo-inositol absorption when orally administered with α-lactalbumin can be beneficial in non-responder patients. Preliminary in vitro findings suggest that peptides deriving from α- lactalbumin digestion may modulate tight junction permeability allowing increased absorption of myoinositol.

Summary The aim of the article was to study the effect of different growth promoters on growth performance, carcass quality, biochemical and haematological traits and immune response of growing rabbits. A total number of 105 male growing NZW rabbits during 35–91 days of age were randomly distributed among 7 groups fed the same basal diet and submitted to different dietary treatments: the first group was unsupplemented and used as control; the other groups were supplemented respectively with bee pollen and/or propolis at 200 mg/kg BW of each and inulin and/or mannanoligosaccharides (MOS) at 35 mg/kg BW of each. Body weight gain, daily feed intake, feed conversion ratio (FCR), biochemical and haematological blood parameters, carcass characteristics, histological studies of ileum and spleen and economical efficiency were measured. Bee pollen administered alone or with propolis significantly (p < 0.01) the body weight gain and improved FCR in respect of the control group. Inulin with MOS significantly improved just FCR than the control group. Bee pollen with propolis and MOS supplemented-groups resulted in significantly higher (7.96 and 8.41% respectively) white blood cells compared to the control group. Plasma total cholesterol was significantly higher for the MOS group in respect of the control, bee pollen, inulin and inulin with MOS supplemented-groups. Propolis resulted in significantly higher dressed carcass percentage than the control group and higher carcass index than only bee pollen with propolis supplemented-group. Bee pollen, in particular if in combination with propolis, could be used as a supplement in the growing rabbits during days 35–91 of age with positive effects on growth rate and feed conversion ratio.

Short Retraction Notice The paper does not meet the standards of "Food and Nutrition Sciences". This article has been retracted to straighten the academic record. In making this decision the Editorial Board follows COPE's Retraction Guidelines. The aim is to promote the circulation of scientific research by offering an ideal research publication platform with due consideration of internationally accepted standards on publication ethics. The Editorial Board would like to extend its sincere apologies for any inconvenience this retraction may have caused. Editor guiding this retraction: Prof. Dr. Alessandra Bordoni (EiC of FNS) The full retraction notice in PDF is preceding the original paper, which is marked "RETRACTED".

Polycystic ovary syndrome (PCOS), a relevant cause of infertility, is a heterogeneous, endocrine disorder affecting up to 10-15% of women in reproductive age. Besides hyperandrogenism, insulin resistance (IR) plays a key role in such syndrome. Insulin-sensitizing drugs, such as Metformin, are effective in treating hyper-insulinemic PCOS patients. Recently, inositols - myo-inositol (MI) and D-chiro-inositol (DCI) - have shown to be an efficient and safe alternative in PCOS management, as both inositol isoforms are able to counteract downstream consequences of insulin resistance. Yet, whereas DCI contributes in mediating insulin activity mainly on non-ovarian tissues, MI displays specific effects on ovary, chiefly by modulating glucose metabolism and FSH-signaling. Moreover, MI may also improve ovarian functions by modulating steroid metabolism through non-insulin-dependent pathways. As DCI and MI activity likely involves different biological mechanisms, both inositol isoforms can be synergistically integrated according to a multitargeted design, by combining MI and DCI in a ratio corresponding to their physiological plasma relative amount (40:1). New experimental and clinical evidence with MI plus DCI evidenced the suitability of such integrated approach, and provided promising results. Further studies need to investigate thoroughly the molecular mechanism and confirm such preliminary data.

This study evaluated the gut and peripheral immune response to genetically modified (GM) maize in mice in vulnerable conditions. Weaning and old mice were fed a diet containing MON810 or its parental control maize or a pellet diet containing a GM-free maize for 30 and 90 days. The immunophenotype of intestinal intraepithelial, spleen, and blood lymphocytes of control maize fed mice was similar to that of pellet fed mice. As compared to control maize, MON810 maize induced alterations in the percentage of T and B cells and of CD4(+), CD8(+), gammadeltaT, and alphabetaT subpopulations of weaning and old mice fed for 30 or 90 days, respectively, at the gut and peripheral sites. An increase of serum IL-6, IL-13, IL-12p70, and MIP-1beta after MON810 feeding was also found. These results suggest the importance of the gut and peripheral immune response to GM crop ingestion as well as the age of the consumer in the GMO safety evaluation.

The commercialisation of Bt (Bacillus thuringensis) maize expressing the Cry endotoxin specific for some types of Lepidopteran and Coleopteran pests has lead researchers to study the reduction in mycotoxin concentrations in Bt hybrids compared to the correspondent isogenic plants. Indeed, insect damage is one of the main inoculation pathways of mycotoxinogenic moulds. The present study aims to evaluate, according to the scientific literature published to date, the real efficiency of Bt maize hybrids in reducing the mycotoxin problem. The results obtained from the analysis of the literature do not show significant variations in the content of aflatoxins, zearalenone and trichothecenes, between Bt hybrids and corresponding isogenic control plants. The only mycotoxins where Bt hybrids have any effect are the fumonisin group, but even in this case studies in commercially planted fields have shown that their effect is mitigated by many biotic and abiotic factors.

Cloricromene decreases myocardial infarct size after ischemic-reperfusion injury in vivo, and it has been suggested that this is due to inhibition of tumor necrosis factor-alpha (TNF-alpha). The purpose of this work was to characterize the mechanism of cloricromene-induced inhibition of TNF-alpha in rat macrophages. Cloricromene inhibited lipopolysaccharide-induced TNF-alpha release in a dose-dependent manner (IC(50)=5.9 +/- 0.8 microM). This was not due to cytotoxicity, as cloricromene was well tolerated up to 500 microM. Cloricromene inhibited lipopolysaccharide-induced expression of TNF-alpha mRNA, which suggests a pre-transcriptional effect. We then investigated the early signal transduction pathway triggered by lipopolysaccharide. The binding of lipopolysaccharide to its receptor CD14 activates protein kinase C and nuclear factor-kappaB (NF-kappaB). Cloricromene inhibited NF-kappaB activation in a dose-dependent manner, but affected protein kinase C translocation only slightly. We then established that cloricromene inhibited lipopolysaccharide-induced cellular oxidative activity, which is important for NF-kappaB activation. Our results show that cloricromene interferes with the early signal transduction pathway triggered by lipopolysaccharide.

The effects of different dietary amounts of organic and inorganic Zn were studied in male White Pekin ducks (WPD) from 1 to 56 days of age. The control diet (26 ppm of Zn from raw ingredients) was supplemented with 30, 60 and 120 ppm of Zn from both inorganic and organic sources, for a total of seven treatment groups, each containing five replicates of nine 1-day-old ducklings each. BW, feed intake and feed conversion ratio (FCR) were recorded at 1, 28 and 56 days of age. At 56 days of age, five birds per group were used in a digestibility trial to measure Zn retention and excretion. At the end of the trial, five birds per treatment were slaughtered and carcass traits as well as Zn content in tibia and liver were measured. Samples of blood from five birds per treatment were used to measure plasma concentration of Zn and Cu. BW gain during the entire period of the trial increased (P < 0.001) by 30 and 60 ppm of Zn. Increasing Zn contents progressively increased (P < 0.001) the tibia and the liver Zn contents as well as the plasma Zn and Cu contents. The concentration of 120 ppm of Zn increased (P < 0.001) tibia ash and decreased (P < 0.001) abdominal fat in the carcasses. In the period 1 to 56 days, Zn oxide increased (P < 0.001) growth rate and improved (P < 0.03) FCR compared with organic Zn, whereas organic Zn increased (P < 0.003) the dressed carcass percentage. Organic Zinc increased (P < 0.001) Zn and Cu concentration in the plasma. A level of 30 ppm of Zn from an inorganic source was adequate for male WPD during 1 to 56 days of age, based on positive effects of growth rate and Zn excretion.

The synthetic analogue of phosphatidylserine, cholesterylphosphorylserine (CPHS) inhibits T-cellmediated immune responses in mice. Tested in cultured mouse spleen cells, CPHS inhibits concanavalin A-induced activation of DNA synthesis ( IC 50 , 3.5 μM). Injected i.p. during the efferent phase, CPHS (25–100 mg/kg) inhibits the manifestations of delayed-type of hypersensitivity. The compound (25 mg/kg i.p., daily) reduces the acute graft-versus-host reaction when given for 5 days to donor mice before the isolation of spleen cells used for the inoculum. These data suggest that the addition of a phosphorylserine group to a steroid ring may produce immunoregulatory compounds.

Since the central nervous system and neuropeptides modulate immune functions, we investigated whether the different susceptibility of Lewis and Brown Norway rats to experimental allergic encephalomyelitis could also reflect differences in beta-endorphin and substance P concentrations in brain areas and macrophages during the development of the disease. We show that beta-endorphin concentrations increase much more in the hypothalamus and macrophages of Lewis rats during the development of the disease, while the increase is much lower or absent in Brown Norway rats. Tumor necrosis factor-alpha seems to play an important role in this difference. The administration of the opiate receptor antagonist naltrexone worsens the development of the disease, suggesting that the increase of the opioid beta-endorphin might represent a mechanism to downregulate the immune response. In both strains, the concentrations of substance P do not change.

Adrenaline has been shown to inhibit the release of tumor necrosis factor (TNF) elicited by lipopolysaccharide (LPS) when tested in vitro on cultured human blood cells and rat macrophages. In this report we have examined the effect of the in vivo administration of adrenaline on TNF serum levels induced by LPS. In agreement with in vitro data, adrenaline (0.1 mg/kg, s.c.) was found to inhibit in the mouse the LPS-induced TNF release. The β-adrenergic antagonist propranolol administered 1 h before adrenaline completely blocked the adrenaline activity, whereas the α-adrenergic antagonist phentolamine was ineffective. These data demonstrate that: (i) adrenaline is an effective antagonist of LPS-induced TNF release in vivo, and (ii) its effect is mediated by β-adrenergic receptors.

Lycopene β-cyclase (tlcy-b) tomatoes, obtained by modulating carotenogenesis via genetic engineering, contain a large amount of β-carotene, as clearly visible by their intense orange colour. In the present study we have subjectedtlcy-btomatoes to anin vitrosimulated digestion and analysed the effects of digestate on cell proliferation. To this aim we used HT-29 human colon adenocarcinoma cells, grown in monolayers, as a model. Digested tomatoes were diluted (20 ml, 50 ml and 100 ml/l) in culture medium and added to the cells for different incubation times (24 h, 48 h and 72 h). Inhibition of cell growth by tomato digestate was dose-dependent and resulted from an arrest of cell cycle progression at the G0/G1 and G2/M phase and by apoptosis induction. A down-regulation of cyclin D1, Bcl-2 and Bcl-xl expression was observed. We also found that heat treatment of samples before digestion enhanced β-carotene release and therefore cell growth inhibition. To induce with purified β-carotene solubilised in tetrahydrofuran the same cell growth inhibition obtained with the tomato digestate, a higher amount of the carotenoid was necessary, suggesting that β-carotene micellarised during digestion is utilised more efficiently by the cells, but also that other tomato molecules, reasonably made available during digestion, may be present and cooperate with β-carotene in promoting cell growth arrest.

The genetic modification in fruit and vegetables could lead to changes in metabolic pathways and, therefore, to the variation of the molecular pattern, with particular attention to antioxidant compounds not well-described in the literature. The aim of the present study was to compare the quality composition of transgenic wheat ( Triticum durum L.), corn ( Zea mays L.), and tomato ( Lycopersicum esculentum Mill.) to the nontransgenic control with a similar genetic background. In the first experiment, Ofanto wheat cultivar containing the tobacco rab1 gene and nontransgenic Ofanto were used. The second experiment compared two transgenic lines of corn containing Bacillus thuringiensis "Cry toxin" gene (PR33P67 and Pegaso Bt) to their nontransgenic forms. The third experiment was conducted on transgenic tomato ( Lycopersicum esculentum Mill.) containing the Agrobacterium rhizogenes rolD gene and its nontransgenic control (cv. Tondino). Conventional and genetically modified crops were compared in terms of fatty acids content, unsaponifiable fraction of antioxidants, total phenols, polyphenols, carotenoids, vitamin C, total antioxidant activity, and mineral composition. No significant differences were observed for qualitative traits analyzed in wheat and corn samples. In tomato samples, the total antioxidant activity (TAA), measured by FRAP assay, and the naringenin content showed a lower value in genetically modified organism (GMO) samples (0.35 mmol of Fe (2+) 100 g (-1) and 2.82 mg 100 g (-1), respectively), in comparison to its nontransgenic control (0.41 mmol of Fe (2+) 100 g (-1) and 4.17 mg 100 g (-1), respectively). On the basis of the principle of substantial equivalence, as articulated by the World Health Organization, the Organization for Economic Cooperation and Development, and the United Nations Food and Agriculture Organization, these data support the conclusion that GM events are nutritionally similar to conventional varieties of wheat, corn, and tomato on the market today.

In the present study, we utilised an in vitro digestion procedure to deliver molecules contained in tomatoes to cultured cells and to analyse potential mechanisms underlying the antitumoural effects of tomatoes reported in the literature. Ripe tomatoes underwent in vitro simulated digestion and the aqueous fraction obtained was delivered to HT-29 and HCT-116 colon adenocarcinoma cells. The amount of lycopene released during digestion and transferred to the aqueous fraction during digestion was 10-fold lower than that present in tomato homogenate before digestion. The carotenoid was accumulated by colon adenocarcinoma cells in a dose-dependent manner after the addition of tomato digestate (20 –100 ml/l) for 24 h. Tomato digestate inhibited the growth of HT-29 and HCT-116 cells in a dose-dependent manner. Growth inhibition resulted from an arrest of cell cycle progression at the G0/G1 phase and by apoptosis induction. A down regulation of cyclin D1, Bcl-2 and Bcl-xL expression was also observed, without apparent changes in p53, p21, p27 and Bax. In conclusion, the present data demonstrate that the in vitro digestion procedure represents a useful approach to supply tomato to colon cultured cells. Moreover, we have shown that tomato digestate is able to inhibit the growth of colon cancer cells by modulating the expression of regulators of the cell cycle and apoptosis.

GMO crops were introduced for commercial production in 1996. Since then, their use has increased rapidly. GMOs have primarily benefited large farms and multinational companies in Industrialised Countries and now is more and more debating their utilisation in Developing World. The objective of the present review is an analysis of this subject from a comprehensive point of view; in addition to that, the changes related to the nutritional content of transgenic foods will be treated. Despite the progress that has been made, the world food situation is still marked by mass hunger and chronic malnutrition. In particular micronutrient malnutrition, that means vitamin and mineral deficiencies, represents an important public health problem in several areas of the world. The "golden rice" bioengineered to contain beta-carotene, as a source of vitamin A is the most famous example of GM food used for reduction (or even to solve) of a public health problem. The expected results of this approach have presently not been achieved. Further studies are necessary to increase the general knowledge about GMOs and their long-term effects on human health. Collaborative attitude of different research sectors (private and public) and involvement of different sectors of society will be an added value for comprehension of the real impact of the application of modern biotechnology to food and agriculture systems.

The synthetic steroid cholesterylphosphoserine (CPHS) inhibited the secretion of TNF-alpha in lipopolysaccharide-challenged human monocytes. CPHS (5-20 microM) was effective when added together with the endotoxin, or after an interval (1-2 h) sufficient to have allowed for initiation of TNF-alpha synthesis. Consistently, CPHS did not alter TNF-alpha gene transcription. In contrast to its action on TNF-alpha, CPHS showed only marginal effects on interleukin 1beta secretion. Given intraperitoneally to mice 2 h before lipopolysaccharide CPHS prevented the rise in plasma TNF-alpha (IC(50): 5 mg/kg). The inhibition of TNF-alpha secretion by CPHS may contribute to the immunosuppressive activity of this steroid.

In human cultured monocytes we examined the ability of myelin basic protein (MBP) to induce the production of proinflammatory cytokines potentially involved in inflammatory demyelination. Northern blots and specific immunoassays demonstrated that monocytes incubated with optimal doses of MBP showed increased mRNA expression and release of tumor necrosis factor (TNF-alpha), interleukin-1beta (IL-1beta), interleukin-6 (IL-6), interleukin-8 (IL-8) but not of interleukin-12/p40 (IL-12/p40). We also showed that cytokine production by MBP-stimulated monocytes was abrogated by incubation with Dexamethasone. These data suggest that interaction of mononuclear phagocytes with MBP may participate in the regulatory process of cytokine production during inflammatory demyelination and support the beneficial role of corticosteroids therapy in aberrant immune responses to the myelin sheath.

Following pre-treatment with a non-depleting anti-CD4 mAb (H129.19) that produces long-lasting receptor saturation, PL/J mice were fully protected from experimental auto-immune encephalomyelitis (EAE) induced by injection of myelin basic protein (MBP). These mice did not develop EAE following MBP re-challenge 5-10 weeks later when the CD4+ cells were no longer coated by the mAb and their lymph node cells were specifically unresponsive to MBP stimulation in vitro. Moreover, superantigen staphylococcal enterotoxin B (SEB) inoculation, which re-induces EAE in MBP immunized mice, failed to activate encephalitogenic T-cells in anti-CD4 + MBP treated mice, even after MBP re-challenge, indicating that tolerance in the peripheral T-cell compartment was achieved. However, MBP re-challenge 16 weeks later, but not SEB, produced an acute episode of EAE in these mice, while it failed to induce disease in a parallel group of adult thymectomized mice. These results indicate that no memory of the first priming exists at this time and that new MBP-specific T-cell precursors are peripheralized and produce EAE after MBP recognition.

We tested the effect of bovine cortex phosphatidylserine (BC-PS), a membrane phospholipid known to inhibit the release of the cytokine tumor necrosis factor (TNF), in SJL/J mice sensitized for adoptively transferred experimental autoimmune encephalomyelitis (EAE). Control, sensitized mice developed severe clinical and histologic EAE within 6 to 9 days, whereas only 20% of mice given BC-PS displayed clinical signs that were much less severe and minimal CNS pathology. Cessation of BC-PS treatment after 15 to 40 days led to disease within 1 to 2 weeks, but when treatment was more prolonged, animals remained healthy after cessation. Serial transfer of spleen cells (SO from BC-PS-treated and control animals into naive recipients resulted in acute EAE within 6 to 7 days with cells from either donor type. Animals treated late with BC-PS failed to relapse and generally remained healthier than controls did over a 40-day period of observation. Cultures of lymph node cells or SC from BC-PS-treated and control animals showed an 80 to 90% reduction in TNF production in the BC-PS-treated group. Thus, we demonstrate that PS can abrogate or significantly reduce the severity of EAE without permanently inhibiting effector T cells. The approach might be considered a candidate for future therapies of relevance to MS.

The hypothesis of this study is summarized in Fig. 6. Phosphatidylserine due to distribution in the internal side of plasma membrane is prevented to react with the extracellular environment. When injury to cell occurs, phospholipid asymmetry is lost and the exposed phosphatidylserine becomes a signal of cell damage. Phosphatidylserine may activate defense reactions while it is still anchored to plasma membrane (Zwaal, 1978; Tanaka and Schroit, 1983). Alternatively, the soluble lysophosphatidylserine is generated, ready to diffuse and transmit the information of tissue damage to other cells. In this sequence of events, lysophosphatidylserine becomes an autacoid, originated from a membrane phospholipid. In rodents, lysophosphatidylserine seems specifically devoted to activate mast cells. The role of these cells in the regulation of the immune reactions and in tissue repair has been advocated (Dexter et al., 1981). The lysophosphatidylserine-induced mast cell activation has been shown in vivo and in vitro in a variety of rodent species (mouse, rat, gerbil, hamster). It may occur through a direct effect or through the participation of synergistic endogenous compounds. Structure-activity relationships in the action of lysophosphatidylserine show that the effect on mast cells is linked to a definite molecular organization. Determinants of the mast cell activation are the free amino group and the carboxyl group of the serine. Support to the general hypothesis of this study originates from the observation that active lysophosphatidylserine is generated within a population of leukocytes, the cells migrating in areas of wounded tissue (Mietto et al., 1987). Production of lysophosphatidylserine can be anticipated in pathological situations associated with extensive cell death (tumor growth, graft rejection, burns). At present, the observations on lysophosphatidylserine are confined to rodent mast cells. Other histamine-secreting cells (e.g., the human basophil) are unresponsive to this phospholipid (Kolster et al., 1987). Among the endogenous compounds interacting with lysophosphatidylserine, nerve growth factor seems of particular interest (Bruni et al., 1982). The synergism with lysophosphatidylserine has been confirmed in other laboratories (Sugiyama et al., 1985; Pearce and Thompson, 1986; Mazurek et al., 1986). The concerted effects by these two compounds on mast cells is in line with current opinion on the participation of nerve growth factor in the regulation of inflammatory and immune reactions (Mietto et al., 1987; Weskamp and Otten, 1987).(ABSTRACT TRUNCATED AT 400 WORDS)