The management of hepatocellular carcinoma. Current expert opinion and recommendations derived from the 24th ESMO/World Congress on Gastrointestinal Cancer, Barcelona, 2022.

This article summarises expert discussion on the management of patients with hepatocellular carcinoma (HCC), which took place during the 24th World Gastrointestinal Cancer Congress (WGICC) in Barcelona, July 2022. A multidisciplinary approach is mandatory to ensure an optimal diagnosis and staging of HCC, planning of curative and therapeutic options, including surgical, embolisation, ablative strategies, or systemic therapy. Furthermore, in many patients with HCC, underlying liver cirrhosis represents a challenge and influences the therapeutic options.

Host conservatism, host shifts and diversification across three trophic levels in two Neotropical forests.

Host-parasite systems have been models for understanding the connection between shifts in resource use and diversification. Despite theoretical expectations, ambiguity remains regarding the frequency and importance of host switches as drivers of speciation in herbivorous insects and their parasitoids. We examine phylogenetic patterns with multiple genetic markers across three trophic levels using a diverse lineage of geometrid moths (Eois), specialist braconid parasitoids (Parapanteles) and plants in the genus Piper. Host-parasite associations are mapped onto phylogenies, and levels of cospeciation are assessed. We find nonrandom patterns of host use within both the moth and wasp phylogenies. The moth-plant associations in particular are characterized by small radiations of moths associated with unique host plants in the same geographic area (i.e. closely related moths using the same host plant species). We suggest a model of diversification that emphasizes an interplay of factors including host shifts, vicariance and adaptation to intraspecific variation within hosts.

Effect of zinc supplementation on vitamin status of middle-aged and older European adults: the ZENITH study.

OBJECTIVE: To assess the effects of zinc supplementation on vitamin status in middle-aged and older volunteers. SUBJECTS/METHODS: Three hundred and eighty-seven healthy middle-aged (55-70 years) and older (70-85 years) men and women, randomly allocated to three groups to receive 15 or 30 mg Zn/day or placebo for 6 months. Dietary intake was assessed by means of a validated 4-day recall record. Fasting blood samples were simultaneously analysed for levels of plasma retinol and alpha-tocopherol by high-performance liquid chromatography. Erythrocyte folates were measured by a competitive immunoassay with direct chemiluminescence detection on an automatized immunoanalyser. Biochemical measurements were performed at baseline and after 3 and 6 months of zinc supplementation. RESULTS: Plasma vitamin A levels were significantly increased proportionally with zinc dose and period of treatment, particularly at 6 months (for 15 mg Zn/day, P<0.05; for 30 mg Zn/day, P<0.0001); no significant changes were observed in the placebo group. There was no effect of zinc supplementation on vitamin E/cholesterol ratio and erythrocyte folates. CONCLUSIONS: Our results show that a long-term zinc supplementation increases plasma vitamin A levels in middle-aged and older people of similar characteristics to those involved in this study. Moreover, supplementation influences serum zinc levels but does not affect erythrocyte zinc concentration and both plasma vitamin E and erythrocyte folate status.

Importance of zinc in the elderly: the ZENITH study.

The elderly are at nutritional risk as a result of multiple physiological, social, psychological, and economic factors. Physiological functions naturally decline with age, which may influence absorption and metabolism. Social and economic conditions can adversely affect dietary choices and eating patterns. However, at the same time, the nutrient needs of the elderly for certain nutrient (such as vitamins, minerals, proteins) is higher than for younger adults. This article reviews the importance of zinc (Zn) in elderly people, particularly for behavioural and mental function, micronutrient status, immune and antioxidant system, and bone metabolism.

Estimation of intake and status of vitamin A, vitamin E and folate in older European adults: the ZENITH.

OBJECTIVE: To report selected dietary intake and vitamin status at baseline of volunteers participating in the ZENITH study and the correlation of vitamin status with zinc. DESIGN: A multicentre prospective intervention study employing a randomised double-blind design. SETTING: Clermont-Ferrand, Theix (France), Coleraine (Northern Ireland), Grenoble (France), Rome (Italy). PARTICIPANTS: In total, 387 healthy middle-aged (55-70 y) and older (70-87 y) men and women participated in the study. METHODS: Dietary intake was assessed by means of a validated 4-d recall record. Fasting blood samples were simultaneously analysed for retinol and alpha-tocopherol by the HLPC method. Erythrocyte folates were measured by a competitive immunoassay with direct chemiluminescence detection on an automatised immunoanalyser. RESULTS: In all centres, men had a significantly (P < 0.0001) higher mean nutrient intake than women. Comparison between age-groups showed that older individuals had significantly lower intakes of macro- and selected micronutrients than middle-aged subjects (P < 0.0001). A high fat intake (from 36 to 40% of total energy) was observed in all examined groups. In relation to biochemical measures of vitamin status, all parameters were above their respective cut-off values for normality and, thus, none of the subjects had biochemical evidence of deficiency of these selected vitamins. A moderate correlation was found with plasma vitamin A and serum zinc (r = 0.12, P < 0.05) or red blood cell zinc (r = 0.12, P < 0.01) and with erythrocyte folates and red blood cell zinc (r = 0.11, P < 0.05). CONCLUSIONS: There were only moderate differences in the nutrient intake of the ZENITH study volunteers among the four European centres. Their biochemical status for retinol, alpha-tocopherol and folate appeared adequate.

Introduction to the ZENITH study and summary of baseline results.

Zinc is known to be essential for a great number of biochemical activities and physiological and cognitive functions. The objective of Zenith study was to investigate the effects of Zn, as a nutritional supplement, on psychological and behavioural factors and on surrogate markers that are indicative of trends towards better health in order to evaluate the need for dietary recommendations specific to the studied population. We report in this paper the summary of baseline results obtained before Zn supplementation.

Basal metabolic rate and thyroid hormones of late-middle-aged and older human subjects: the ZENITH study.

OBJECTIVE: This paper describes baseline data on basal metabolic rate (BMR), thyroid hormone levels and body composition of middle-aged and older people participating in the ZENITH project and the correlation of thyroid hormone levels with zinc status. DESIGN: A multicentre prospective intervention study employing a randomised double blind design. SETTING: Clermont-Ferrand, Theix (France), Coleraine (Northern Ireland), Grenoble (France), Rome (Italy). INTERVENTIONS: BMR has been measured on a subsample of 70 middle-aged volunteers (35 men and 35 women recruited in Clermont-Ferrand, France, aged 55-70 y) and 108 older volunteers (56 men and 52 women recruited in Rome, Italy, aged 70-85 y). Thyroid hormone levels were evaluated in the entire group of ZENITH volunteers (n = 387). BMR was measured by indirect calorimetry. Fat-free mass (FFM) was derived by four skinfold thicknesses using Durnin and Womersley's equations. Concentrations of thyroid hormones (total T3 and T4) were measured using a competitive immunoassay with an enhanced chemiluminescence end point. RESULTS: Italian older volunteers had a significantly lower FFM than middle-aged French volunteers (-7% P < 0.01). A negative correlation between BMR and age (men, r = -0.64; women, r = -0.62; both P < 0.0001) was observed: BMR was significantly (P < 0.000001) lower in Italian elderly volunteers (4.03+/-0.46 kJ/min and 3.29+/-0.42 kJ/min for men and women, respectively) than in middle-aged French volunteers (4.84+/-0.45 kJ/min and 3.87+/-0.38 kJ/min for men and women, respectively), even after adjustment for FFM (-12%). No correlation has been observed between BMR and thyroid hormones both in French and Italian subjects. Total T4 (TT4) concentrations were lowest in middle-aged population (-10%, P < 0.0001). A moderate negative correlation has been found with TT4 and red blood cell zinc (r = -0.12, P < 0.02, slope -0.026). CONCLUSIONS: The results confirm an age-related decline in BMR not entirely explained by body composition or thyroid hormones differences.

Zinc status and age-related changes in peripheral blood leukocyte subpopulations in healthy men and women aged 55-70 y: the ZENITH study.

OBJECTIVE: To determine zinc status and age-related changes in the immune function of healthy late-middle-aged men and women (aged 55-70 y). DESIGN: Observational study. SETTING: Population of Northern Ireland. SUBJECTS: Apparently healthy, free-living individuals (45 men, 48 women) aged 55-70 y. INTERVENTION: Zinc status markers were analysed by flame atomic absorption spectrometry and commercially available kits. Immune function was assessed by flow cytometry. RESULTS: Serum and erythrocyte zinc concentrations were 13.0 (s.d. 1.40) micromol/l and 222 (s.d. 48.2) micromol/l, respectively. Serum alkaline phosphatase (ALP) concentrations were 76.8 (s.d. 16.1) U/l; women showed significantly higher concentrations of ALP (P = 0.011). Women demonstrated (1) a significant inverse correlation in naive T lymphocytes, specifically naive T-helper lymphocytes (% expression, r = -0.364, P = 0.007 and absolute count, r = -0.275, P = 0.036) with age and (2) a significant positive correlation between late activation of T lymphocytes (% expression, r = 0.299, P = 0.019 and absolute count, r = 0.260, P = 0.039) with advancing age. Men demonstrated a significant positive correlation in the % expression of (CD3-/CD16+/CD56+) natural killer (NK) cells with age (r = 0.316, P = 0.017). CONCLUSIONS: Between the ages of 55 and 70 y, healthy individuals experience significant alterations in immune function; however, such changes appear largely sex specific. Given the reported importance of adequate zinc status in maintaining optimal immune function, further studies are required to explore the effect of enhanced zinc status on emerging immune deficiencies in cell-mediated immunity in healthy 55-70 y olds.

Age-related oxidative stress and antioxidant parameters in middle-aged and older European subjects: the ZENITH study.

OBJECTIVE: Oxidative stress has been reported to increase with ageing. However, the data in healthy humans remain controversial and studies in free-living elderly people are scarce. The objective of the present study was to compare age-related oxidative stress in late middle-aged and older free-living subjects. DESIGN: The effect of ageing on oxidative stress and antioxidant parameters was investigated in 188 middle-aged subjects from Clermont-Ferrand (France) and Coleraine (UK), and in 199 older subjects from Grenoble (France) and Roma (Italy). Plasma thiol (SH) groups, define definition (TBAR's) and total glutathione (GSH), define definition (FRAP), and superoxide dismutase (SOD) activity were measured at baseline of the ZENITH study. RESULTS: Plasma SH groups and FRAP and, surprisingly, TBAR's were significantly lower in free-living older subjects compared to younger subjects (P < 0.001, P < 0.001, P < 0.01, respectively), but there was no significant differences in GSH levels. CONCLUSION: European free-living healthy older do not appear to be exposed to an acute oxidative stress. However, the highly significant positive correlation between plasma SH group oxidation or decreased FRAP and ageing is predictive of an increased risk of oxidative stress in older subjects. Moreover, the comparison between middle-aged and older subjects regarding oxidative stress parameters suggests also a progressive and slow decline of antioxidant status in healthy free-living older elderly and underline the impact on life-style factors on successful ageing.

The relationship between the zinc nutritive status and biochemical markers of bone turnover in older European adults: the ZENITH study.

OBJECTIVE: To investigate the relationship between indices of zinc nutritive status and biochemical markers of bone turnover in older adult European subjects. DESIGN: Use of baseline data from a multicentre prospective zinc intervention (ZENITH) study. SETTING: Centres in France, Italy and Northern Ireland. PARTICIPANTS: A total of 387 healthy adults, aged 55-87 y. METHODS: Zinc intake was assessed by 4-day recall records. Circulating and urinary biochemical zinc status measures were assessed by atomic absorption spectrophometry. Serum bone-specific alkaline phosphatase and osteocalcin were assessed by ELISA and urinary pyridinoline (Pyr) and deoxypyridinoline (Dpyr) by HPLC. RESULTS: Zinc intake was negatively correlated with urinary Pyr and Dpyr (r = -0.298 and -0.304, respectively; P < 0.0001), but was not correlated with bone formation markers. There was a tendency for serum zinc to be negatively correlated with urinary Dpyr (r = -0.211; P = 0.080). Erythrocyte zinc was negatively correlated with serum osteocalcin (r = -0.090; P < 0.0001). None of the other correlations were significant. After adjustment for confounder (age, gender and research centre) the only significant association that remained was between serum osteocalcin and erythrocyte zinc (beta = -0.124; P = 0.011). CONCLUSIONS: There was some, albeit inconsistent, evidence of a relationship between zinc nutritive status and bone turnover in the older adult participants of the ZENITH study.

Positive and negative mood in the elderly: the ZENITH study.

OBJECTIVE: To assess the quality of positive and negative affect (mood) in an ageing European sample. BACKGROUND: Mood quality has important implications for both physical and mental wellbeing. Poor quality moods are associated with deficits in the diverse areas of cognitive function, health, and social relationships. The ageing process presents a number of potential challenges to successful mood regulation that could have wider implications. DESIGN AND PARTICIPANTS: The current study examines the quality of positive and negative affect in 387 healthy participants from three European countries. Moods were measured four times a day for 4-7 d with the Positive and Negative Affect Schedule (PANAS) mood scales. Measures of zinc (Zn) status were taken also. SETTING: Two centres concentrated on 55-70 yr olds (Coleraine, N.Ireland, n = 93 and Clermont-Ferrand, France, n = 95), and two centres concentrated on 70-87 yr olds (Rome, Italy, n = 108, and Grenoble, France, n = 91). RESULTS: Positive affect scores for the centre in Rome were significantly (P < 0.01) lower than for the other three centres, and the Grenoble centre had significantly (P < 0.05) higher scores on negative affect than the other three centres. Mood was not related to measures of zinc status (all Ps > 0.05). CONCLUSIONS: The two centres with the oldest participants showed deficits in mood quality that may have implications for broader well-being.

Zinc status and taste acuity in older Europeans: the ZENITH study.

BACKGROUND: Age-related decline in taste acuity may be both a cause and an effect of depleted zinc and/or increased zinc requirement. OBJECTIVE: The aim of this study was to explore associations between zinc status and taste acuity in healthy older European adults aged 55-90 y. SAMPLE: Volunteers were recruited within Italy (n = 108 aged 70-90 y), the United Kingdom (UK) (n = 93 aged 55-70) and two regions of France (n = 186), Grenoble (aged 70-90 y) and Clermont-Ferrand (aged 55-70 y). METHODS: A signal detection theory approach was adopted, employing a three-alternative, forced-choice procedure. The data were converted to R-indices and bivariate correlations were computed to explore relationships between serum zinc, erythrocyte zinc and taste acuity. ANOVA was undertaken to determine regional differences in zinc status. RESULTS: Higher erythrocyte zinc status was associated with better acuity for salt (sodium chloride) taste in the sample as a whole (P = 0.012) (n = 385). Higher serum zinc levels were associated with greater sensitivity to sour taste (citric acid) (P = 0.015) only in the older groups (aged 70-90 y). There were no apparent associations between serum or erythrocyte zinc status and acuity for bitter (quinine) or sweet (sucrose) tastes irrespective of age. CONCLUSION: These results agree with those previously suggesting that age-related detriment in sensitivity for salt taste may be associated with depleted zinc.

Zinc intake and status in middle-aged and older European subjects: the ZENITH study.

OBJECTIVE: Inadequate intakes of micronutrients in elderly negatively affect the nutritional status. Zinc is an essential micronutrient in the elderly, especially in relation to its impact on immune function, bone mass, cognitive function and oxidative stress. However, data are lacking on zinc intake and status during normal ageing. In this study, we evaluate the intake and status of zinc in late middle-aged and older free-living subjects. DESIGN: Dietary zinc intake and zinc status in 188 middle-aged subjects from Clermont-Ferrand (Fr) and Coleraine (UK), and in 199 older subjects from Grenoble (Fr) and Roma (It) were assessed at the entry in the ZENITH study. RESULTS: In relation to the zinc RDA for people older than 55 y, zinc intakes in most of the middle-aged and older subjects (more than 96%) in the present study were adequate. Older people had significantly lower (P < 0.01) energy intakes as compared to middle-aged. Zinc intake expressed per MJ was also significantly (P < 0.01) higher in older people compared to middle-aged. Erythrocyte and urinary zinc concentrations were significantly (P < 0.001) higher in middle-aged subjects compared to older ones. The prevalence of biological Zn deficiency in free-living ageing European people was low (<5%). CONCLUSION: The results of the present study showed a relatively low prevalence of zinc deficiencies in healthy free-living late middle-age and older subjects. These results should be useful for health professionals to have reference data on zinc intake and status for a healthy ageing.

Screening and recruitment procedure of late-middle aged and older subjects: the ZENITH study.

OBJECTIVE: To report the rational, design, recruitment, baseline characteristics and preliminary overview of volunteers in the ZENITH study. DESIGN: A multicentre prospective intervention study employing a randomised double-blind design. SETTING: Clermont-Ferrand, Theix (France), Coleraine (Northern Ireland), Grenoble (France), Rome (Italy). PARTICIPANTS: Healthy men and women middle-aged (55-70 y) and older volunteers (70-87 y). INTERVENTIONS: At baseline (prior to zinc (Zn) supplementation), all volunteers underwent a full clinical examination, anthropometric measurements, health and lifestyle questionnaire, Mini Mental State Examination, Geriatric Depression Scale, and biochemistry profile. RESULTS: In total, 842 volunteers (378 men and 464 women) were invited to take part in the study. A total of 49% of these volunteers were excluded on the basis of inclusion/exclusion criteria. In total, 433 participants were admitted to the Zn supplementation for 6 months. During this period, about 10% of volunteers dropped out from the study. CONCLUSIONS: A total of 387 subjects (197 male and 190 female subjects) successfully completed the supplementation phase of the ZENITH study.

Health and lifestyle characteristics of older European adults: the ZENITH study.

OBJECTIVE: To describe health and lifestyle factors of participants in the ZENITH study. DESIGN: A prospective multicentre intervention study employing a randomised double-blind design. PARTICIPANTS: Community dwelling older adults (n = 387), aged 55-87 y were recruited from regions in France, Italy and the UK. INTERVENTION: A self-report questionnaire comprising socio-demographic variables, dietary habits, physical activity in the home, at work and recreation. RESULTS: Participants differed with regards dietary habits and physical activity for each region. Recreational activity was higher in France and women generally tend to perform less hours of recreational activity per week than men. CONCLUSIONS: The differences found for these regions of Europe in relation to lifestyle factors will affect health and well-being within these countries and may mediate the impact of zinc supplementation on various biological and psychological parameters.

The effect of copper supplementation on red blood cell oxidizability and plasma antioxidants in middle-aged healthy volunteers.

A multicenter European study (FoodCue) was undertaken to provide data on the significance of increased dietary copper as a pro-oxidant or antioxidant in vivo. The present work describes the effect of Cu supplementation on (2,2'-azo-bis(2-amidinopropane) hydrochloride (AAPH)-induced red blood cell oxidation in middle-aged people. Double-blinded copper supplementation was achieved in 26 healthy volunteers (50-70 years) with pills containing 3 mg CuSO(4), 3 mg Cu glycine chelate (CuG) and 6 mg CuG. Each 6 week supplementation period was preceded and followed by 6 weeks of washout (WO) on placebo. The results show significant increases in time necessary to achieve 50% hemolysis (LT(50)) after 3CuSO(4) and 6CuG compared with values after WO periods. Cu supplementation did not increase the levels of (Cu,Zn)SOD activity in red blood cells. Resistance to hemolysis was significantly and positively correlated (r =.30, p <.01) with alpha- and beta-carotene content in the plasma. Together, these data suggest that intake of copper as high as 7 mg/d has no pro-oxidant activity and may rather result in protection of red blood cells against oxidation. The decreased oxidizability of red blood cells did not result from increased (Cu,Zn)SOD activity and may occur through other mechanisms such as changes in membrane antioxidant content.

Copper supplementation in humans does not affect the susceptibility of low density lipoprotein to in vitro induced oxidation (FOODCUE project).

The oxidative modification of low-density lipoprotein cholesterol (LDL) has been implicated in the pathogenesis of atherosclerosis. Copper (Cu) is essential for antioxidant enzymes in vivo and animal studies show that Cu deficiency is accompanied by increased atherogenesis and LDL susceptibility to oxidation. Nevertheless, Cu has been proposed as a pro-oxidant in vivo and is routinely used to induce lipid peroxidation in vitro. Given the dual role of Cu as an in vivo antioxidant and an in vitro pro-oxidant, a multicenter European study (FOODCUE) was instigated to provide data on the biological effects of increased dietary Cu. Four centers, Northern Ireland (coordinator), England, Denmark, and France, using different experimental protocols, examined the effect of Cu supplementation (3 or 6 mg/d) on top of normal Cu dietary intakes or Cu-controlled diets (0.7/1.6/6.0 mg/d), on Cu-mediated and peroxynitrite-initiated LDL oxidation in apparently healthy volunteers. Each center coordinated its own supplementation regimen and all samples were subsequently transported to Northern Ireland where lipid peroxidation analysis was completed. The results from all centers showed that dietary Cu supplementation had no effect on Cu- or peroxynitrite-induced LDL susceptibility to oxidation. These data show that high intakes (up to 6 mg Cu) for extended periods do not promote LDL susceptibility to in vitro-induced oxidation.

An investigation of the biological activity of bullrout (Notesthes robusta) venom.

Bullrout envenomation is known to cause intense pain. Crude bullrout venom and venom fractions were assessed for protease, hyaluronidase, phospholipase and hemolytic activities, reactivity with stonefish antivenom, lethality to brine shrimp and ability to elicit pain in human subjects. Compared with venom obtained from frozen specimens, live fish venom-milking techniques rendered greater venom potency and improved storage characteristics. Although mild proteolytic and hemolytic activity was observed, crude venom demonstrated no hyaluronidase or phospholipase A2 activity, did not affect brine shrimp, or show antigenicity with stonefish antivenom. A single venom protein isolated from bullrout venom is attributed with causing pain in human subjects. The sensations elicited by this novel algesic protein are consistent with chemical stimulation of polymodal nociceptors.

No effect of copper supplementation on biochemical markers of bone metabolism in healthy young adult females despite apparently improved copper status.

OBJECTIVE: To investigate the effects of increasing Cu intakes, above the usual dietary intake, on biomarkers of bone metabolism in healthy young adult females (aged 21-28 y) over a 4 week period. DESIGN: A double-blind, placebo-controlled randomised repeat crossover Cu supplementation trial. SETTING: The study was conducted at the Royal Veterinary and Agricultural University (RVAU), Copenhagen, Denmark. SUBJECTS: Sixteen healthy young adult females aged 20-28 y were recruited from among students at the RVAU. INTERVENTION: During the 4 week intervention periods in this randomised, crossover trial (3x4 weeks with a minimum 3 week wash-out period), each subject received, in addition to their usual diet, either 3 or 6 mg elemental Cu/day as CuSO4 or a matching placebo. On the last 3 days of each dietary period 24 h urines were collected. In addition, blood was collected on the last day of each dietary period. RESULTS: Serum Cu and erythrocyte superoxide dismutase (but not caeruloplasmin protein concentration or activity (putative indices of Cu status)) were significantly increased (P<0.05) after daily Cu supplementation with 3 and 6 mg/day for 4 weeks. Serum osteocalcin (biomarker of bone formation), urinary creatinine (Cr) concentration, urinary pyridinoline (Pyr)/Cr or deoxypyridinoline (Dpyr)/Cr excretion, or daily urinary Pyr or Dpyr excretion (biomarkers of bone resorption) were unaffected by Cu supplementation. CONCLUSION: Copper supplementation of the usual diet in healthy young adult females, while apparently improving Cu status, had no effect on biochemical markers of bone formation or bone resorption over 4 week periods. SPONSORSHIP: Funding from the European Commission.

In vivo DNA/RNA adduction of 7,12-dimethylbenz(a)anthracene (DMBA) and benzo(a)pyrene (BaP) in the liver of rainbow trout (Oncorhynchus mykiss).

Juvenile rainbow trout were exposed via two intramuscular injections to either 14C-DMBA for 24 hr or 14C-BaP for 48 hr, after which the livers were removed for DNA extraction and analysis. In the fish exposed to 14C-BaP, 0.2 ng was bound to the DNA, representing 0.5% of the total liver PAH-derived radioactivity and 2.38% of the administered dose. Liver DNA and RNA were found to contain 0.5% of the administered dose, respectively. Liver analysis of rainbow trout exposed to 14C-DMBA demonstrated that 0.4 ng and 0.3 ng were bound to the DNA and RNA, respectively. This represents 1.0% and 0.6% of the liver DMBA burden, respectively. The DNA adduct concentrations formed were comparable to both in vitro and in vivo experiments with both mammals and fishes, indicating that relatively small, "environmentally realistic" doses of PAH have the ability to bind significantly to critical cellular macromolecules of young fish in vivo.