The likely extinction of hundreds of palm species threatens their contributions to people and ecosystems.

Protecting nature's contributions to people requires accelerating extinction risk assessment and better integrating evolutionary, functional and used diversity with conservation planning. Here, we report machine learning extinction risk predictions for 1,381 palm species (Arecaceae), a plant family of high socio-economic and ecological importance. We integrate these predictions with published assessments for 508 species (covering 75% of all palm species) and we identify top-priority regions for palm conservation on the basis of their proportion of threatened evolutionarily distinct, functionally distinct and used species. Finally, we explore palm use resilience to identify non-threatened species that could potentially serve as substitutes for threatened used species by providing similar products. We estimate that over a thousand palms (56%) are probably threatened, including 185 species with documented uses. Some regions (New Guinea, Vanuatu and Vietnam) emerge as top ten priorities for conservation only after incorporating machine learning extinction risk predictions. Potential substitutes are identified for 91% of the threatened used species and regional use resilience increases with total palm richness. However, 16 threatened used species lack potential substitutes and 30 regions lack substitutes for at least one of their threatened used palm species. Overall, we show that hundreds of species of this keystone family face extinction, some of them probably irreplaceable, at least locally. This highlights the need for urgent actions to avoid major repercussions on palm-associated ecosystem processes and human livelihoods in the coming decades.

Volume of the Sexually Dimorphic Nucleus of the Preoptic Area in Rats Exposed to a Maternal High Fat Diet.

Mice and rats exposed to a maternal diet high in fat have an increased risk of reproductive system tumors later in life. To test whether a part of the hypothalamus associated with sex differentiation, the sexually dimorphic nucleus of the preoptic area (SDN-POA), might be involved, this nucleus was measured in the offspring of female rats on a high, or low fat diet during pregnancy and nursing. The measurements showed that the SDN-POA was smaller in female offspring exposed to a maternal high fat diet than in female offspring exposed to a maternal low fat diet. This could result in endocrine disturbances and thus provide a possible explanation for the linkage between high fat exposure prenatally and increased female reproductive system cancer.

Multigenerational effects of dietary fat carcinogenesis in mice.

The possibility of multigenerational transmission of a carcinogenic effect from exposure to a maternal diet high in fat was tested in mice. Diets with 2.6 or 29% fat (by weight) were fed to strain CD-1 mice during pregnancy. The female offspring were raised on a control diet (10% fat), mated, and continued on the control diet through pregnancy. Their female offspring were raised to terminal illness and autopsied. The total number of reproductive system tumors, pituitary tumors, and metastases was increased in the offspring with ancestral exposure to high dietary fat but to a lesser extent than had been reported previously for direct prenatal exposure to high maternal dietary fat. Because previous work has given evidence against germ cell transmission, a hypothesis based on a maternal effect was offered to explain the multigenerational carcinogenesis. These results have implications for epidemiological studies.

Combination oral antioxidant supplementation reduces blood pressure.

1. Hypertension affects 30% of adults and low intakes of antioxidants have been associated with increased risk of hypertension and cardiovascular disease. To investigate the effect of short-term high-dose antioxidant supplementation on blood pressure in hypertensive and normotensive outpatients, we undertook a randomized, double-blind, crossover design placebo-controlled study. 2. Forty subjects were recruited from medical outpatient clinics, of whom 38 completed the study. Twenty-one were attending for treatment of hypertension and 17 were normotensive, attending for minor gastrointestinal complaints. Subjects were randomly assigned to receive either 8 weeks placebo followed by 2 weeks washout then 8 weeks antioxidants or vice versa. The combination of antioxidants consisted of 200 mg of zinc sulphate, 500 mg of ascorbic acid, 600 mg of alpha-tocopherol (sodium succinate salt) and 30 mg of beta-carotene daily. 3. Systolic blood pressure fell at the end of the antioxidant phase compared with the placebo phase both in subjects receiving anti-hypertensive therapy (P < 0.01) and those who were normotensive (P = 0.067). Circulating levels of beta-carotene and alpha-tocopherol increased in all subjects during supplementation (P < 0.01) and urine nitrite increased in hypertensive patients (P < 0.05). 4. Short-term oral high-dose combination antioxidant therapy reduces blood pressure, possibly via increased availability of nitric oxide. This study may have implications for the innovative use of antioxidants as an adjunct to anti-hypertensive therapy.

Intensity of multigenerational carcinogenesis from diethylstilbestrol in mice.

Mice exposed prenatally to diethylstilbestrol (DES-exposed mice) can transmit a carcinogenic influence to the next generation (DES-lineage mice) when mated to control mice. The persistence of this effect was studied one generation further (DES-lineage-2 mice) by mating DES-lineage female mice to control males. The interaction of maternal dietary fat levels with DES was also tested by feeding high and low levels of dietary fat during the pregnancies that produced the final two generations. DES-lineage-2 mice, exposed to low or high fat maternal diets, had significantly more tumors than control mice with corresponding dietary fat exposure. The frequency of tumors in DES-lineage-2 mice was not significantly lower than in DES-lineage mice from a previous experiment. Thus, the multigenerational effect of DES is relatively intense in mice. If this type of carcinogenesis can occur in the human population, it poses a major threat to future generations.

The effects of intravenous antioxidants in patients with septic shock.

Oxidative stress is implicated in septic shock. We investigated the effect of intravenous antioxidant therapy on antioxidant status, lipid peroxidation, hemodynamics and nitrite in patients with septic shock. Thirty patients randomly received either antioxidants (n-acetylcysteine 150 mg/kg for 30 min then 20 mg/kg/h plus bolus doses of 1 g ascorbic acid and 400 mg alpha-tocopherol) or 5% dextrose. Basal vitamin C was low and redox-reactive iron was elevated in all patients. In the 16 patients receiving antioxidants, vitamin C increased (p = .0002) but total antioxidant capacity was unaffected. Lipid peroxides were elevated in all patients but did not increase further in the patients receiving antioxidants. Plasma total nitrite also increased (p = .007) in the antioxidant group. Heart rate increased in patients receiving antioxidants at 60 min (p = .018) and 120 min (p = .004). Cardiac index also increased at 60 min (p = .007) and 120 min (p = .05). Systemic vascular resistance index decreased at 120 min in the antioxidant treated patients (p = .003). The effect of antioxidants on hemodynamic variables has not previously been reported. Antioxidant administration may be a useful adjunct to conventional approaches in the management of septic shock.

Stage of susceptibility to carcinogenicity of prenatal dietary fat exposure tested by blastocyst transfer.

A previous experiment with exposure of female mice to a diet high in fat from 4 weeks of age through to the end of pregnancy demonstrated a substantial increase in the frequency of reproductive system tumors in their female offspring. Over this wide period of exposure, one or more specific periods of sensitivity to the carcinogenic effects of dietary fat should exist. To test whether a high fat diet affects maternal germ cells and/or zygote development through to blastocyst formation, two blastocyst transfer experiments were performed. Blastocysts from a dam on high fat were transferred to a pseudopregnant dam on low fat. The reverse type of transfer was performed for contrast. These experiments did not reveal any significant effect of dietary fat on maternal germ cells or zygote development.

Congestive cardiac failure following laxative withdrawal.

We report the case of a 60-year-old woman who presented with weakness and hypokalaemia due to excessive use of laxatives. When the laxatives were withdrawn, she developed severe congestive cardiac failure requiring treatment with a diuretic and angiotensin-converting enzyme inhibitor. There was no underlying cardiac abnormality, and these drugs were eventually stopped with no recurrence of the cardiac failure. The possible mechanisms of heart failure following laxative withdrawal is discussed.

Regulation of nitric oxide synthase activity in cultured human endothelial cells: effect of antioxidants.

Nitric oxide release is induced in many cells, including vascular endothelium, as part of the host response to inflammation. Nitric oxide synthase activity is increased in patients with sepsis, associated with increased oxidant demands and decreased antioxidant protection. We used a human vascular endothelial cell line to investigate the influence of antioxidants on nitric oxide synthase activity. Cells were cultured to confluence and incubated with interferon gamma, tumor necrosis factor, and lipopolysaccharide in the combined presence of the antioxidants ascorbic acid, Trolox, catalase, or superoxide dismutase, singly and in combination, for 48 h. Additionally, some cells were incubated with hypoxanthine-xanthine oxidase or a nitric oxide donor. Nitric oxide synthase activity was upregulated by cytokine exposure (p < .0005). Ascorbic acid and superoxide dismutase/ catalase resulted in decreased enzyme activity (p < .05). Superoxide anion release from xanthine oxidase caused increased activity (p < .05) and exogenous nitric oxide tended to suppress synthase activity. We suggest that antioxidants scavenge superoxide anion, enabling feedback inhibition of nitric oxide synthase activity by nitric oxide, and thus reducing enzyme activity. Exogenous nitric oxide also has a similar effect. Superoxide generation suppresses this feedback inhibition. This study has important implications in patients with sepsis in whom nitric oxide synthase inhibitor therapy is currently under investigation.

Effects of fostering on the increased tumor incidence produced by a maternal diet high in fat.

Previous work has shown that female offspring of mice fed a diet high in fat during pregnancy developed more reproductive system tumors and metastases than offspring of pregnant mice fed a low-fat diet. The purpose of the current experiment was to use fostering to test whether the sensitive period for this cancer effect involved the early postnatal period. Strain CD-1 female mice were placed on a diet of 2.6% fat or 29% fat from corn oil at 4 weeks of age and bred at 6-10 weeks of age. The special diets were discontinued at birth, and litters from dams that had been fed the low-fat diet were fostered to dams previously fed the high-fat diet, and vice versa. The offspring were raised to terminal illness and autopsied. There was no difference in age at terminal illness or in the number of the common nonreproductive system tumors between the two fostered groups. Tumor metastases appeared in both groups. However, the combined frequency of reproductive tract tumors and mammary tumors was significantly higher in mice exposed prenatally to a low-fat diet and fostered to dams that had consumed a high-fat diet during pregnancy than in mice exposed prenatally to a high-fat diet and fostered to a dam fed a low-fat diet. Thus the most sensitive period for a cancer effect from high fat was early postnatal, even though the special diets had been discontinued at birth. This matches the period of greatest sensitivity for sex differentiation of the hypothalamus.

Increased reproductive tract tumors in the female offspring of mice fed a high fat diet during the fetal stage of pregnancy.

In a previous study, female offspring of mice consuming a high fat diet throughout pregnancy developed reproductive system tumors and tumor metastases with a frequency significantly higher than offspring of mice on a low fat diet. To test for the sensitive period more specifically, the feeding of a high fat diet was restricted to the fetal period of pregnancy in the present experiment. The offspring were raised to terminal illness and autopsied. The total number of ovarian, uterine and mammary tumors was 14 among 74 mice exposed prenatally to low fat and 34 among 75 mice exposed prenatally to high fat (P < 0.002). In mice exposed to high fat during the fetal period 13 tumors produced metastases, but no metastases were identified after exposure to low fat (P < 0.001). Thus, a maternal diet high in fat during the fetal period of pregnancy was sufficient to increase reproductive system tumors and metastases in the female offspring.

Total antioxidant capacity and lipid peroxidation during liver transplantation.

1. We assessed plasma total antioxidant capacity using enhanced chemiluminescence and plasma lipid peroxides in eight patients undergoing orthotopic liver transplantation and 26 healthy control subjects. 2. The antioxidant capacity of plasma decreased significantly on reperfusion of the donor liver (P < 0.05) during liver transplantation, although levels were not lower in patients than in control subjects. 3. Pre-reperfusion antioxidant capacity correlated negatively with the concentration of lipid peroxidation products appearing in the plasma by the end of the transplant (P < 0.02), but did not correlate with vitamin A or E concentrations. 4. This study provides evidence of oxidant stress during liver transplantation, as shown by antioxidant utilization associated with increased lipid peroxidation.

The effect of anticoagulant choice on apparent total antioxidant capacity using three different methods.

We assessed total antioxidant capacity using three different methods, in plasma samples treated with either EDTA or heparin as anticoagulant, from 26 healthy subjects. Total antioxidant capacity was determined using an oxygen electrode (as the total peroxyl radical-trapping antioxidant parameter), by enhanced chemiluminescence, and by measurement of the antioxidant-mediated quenching of the absorbance of a radical cation. The choice of anticoagulant had a profound effect on antioxidant capacity with heparinized plasma giving consistently higher values than plasma anticoagulated with EDTA. Using the oxygen electrode the mean value was 786.5 +/- 171.5 mumol/L (heparin) compared to 681.4 +/- 160.4 mumol/L (EDTA, P < 0.01). The chemiluminescence technique gave a mean antioxidant capacity of 915.6 +/- 214.1 mumol/L in heparin samples and 714.4 +/- 195.4 mumol/L in EDTA samples (P < 0.0001). The absorbance quenching technique gave a mean value of 867.0 +/- 199.2 mumol/L (heparin) and 675.5 +/- 245.4 mumol/L (EDTA, P < 0.001). All methods tested showed comparable results for EDTA plasma, but the chemiluminescence technique gave higher apparent antioxidant capacity than either of the two techniques when heparin plasma was used. We suggest that either heparin is interacting to enhance antioxidant protection perhaps through release of superoxide dismutase, or the chelation of metal ions by EDTA is limiting the activity of antioxidant metalloenzymes. Consistency in the choice of anticoagulant is clearly extremely important.

Multi-generational carcinogenesis from diethylstilbestrol investigated by blastocyst transfers in mice.

Previous studies have shown that a carcinogenic effect can be transmitted from female mice exposed prenatally to diethylstilbestrol (DES) to their female offspring. Furthermore, male mice exposed pre-natally to DES can transmit a carcinogenic effect to their offspring through their germ cells. To study how multi-generational carcinogenesis is transmitted through females exposed pre-natally to DES, the technique of blastocyst transfer was utilized. Blastocysts from strain CD-1 mice exposed pre-natally to vehicle were transferred to mice exposed pre-natally to DES. Among 143 offspring from these transfers, there were 10 ovarian adenomas and 10 uterine adenocarcinomas. Among 92 offspring from blastocyst transfers between mice exposed pre-natally to vehicle only, there was 1 ovarian adenoma and 1 uterine adenocarcinoma. Thus the pre-natal exposure of the host to DES produced a maternal environment which increased the incidence of ovarian and uterine tumors. The reverse type of transfer was also performed, in which blastocysts from female mice exposed pre-natally to DES were transferred into mice exposed to vehicle only pre-natally. Among 99 offspring derived from DES-exposed germ cells, 6 developed ovarian adenomas and 16 developed uterine adenocarcinomas. Thus DES also has a multi-generational effect transmitted through the blastocyst, which is consistent with fetal germ cell mutation from DES.

Decreased antioxidant status and increased lipid peroxidation in patients with septic shock and secondary organ dysfunction.

OBJECTIVE: To determine antioxidant vitamin concentrations, lipid peroxidation, and an index of nitric oxide production in patients in the intensive care unit (ICU) with septic shock and relate the findings to the presence of secondary organ failure. DESIGN: A prospective, observational study. SETTING: A nine-bed ICU in a University teaching hospital. PATIENTS: Sixteen consecutive patients with septic shock, defined as: a) clinical evidence of acute infection; b) hypo- or hyperthermia (< 35.6 degrees C or > 38.3 degrees C); c) tachypnea (> 20 breaths/min or being mechanically ventilated); d) tachycardia (> 90 beats/min); e) shock (systolic pressure < 90 mm Hg) or receiving inotropes. Fourteen patients also had secondary organ dysfunction. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Antioxidant vitamin concentrations were significantly lower in the patients than the reference range obtained from a comparable group of healthy controls. The mean plasma retinol (vitamin A) concentration was 26.5 +/- 19.3 micrograms/dL compared with 73.5 +/- 18.3 micrograms/dL in healthy subjects (p < .01). Additionally, 13 (81%) patients had retinol values below the lower limit of our reference range (< 37.0 micrograms/dL). Tocopherol (vitamin E) plasma concentrations were below the reference range in all patients (< 9.0 mg/L), with a mean value of 3.6 +/- 2.0 mg/L compared with 11.5 +/- 1.3 mg/L in healthy subjects (p < .001). Plasma beta carotene and lycopene concentrations were undetectable (< 15 micrograms/L) in eight (50%) patients, and below our reference range (< 101 micrograms/L and < 154 micrograms/L, respectively) in the remaining patients. In the five patients with three or more dysfunctional secondary organs, plasma thiobarbituric acid-reactive substances were significantly increased (p < .05), suggesting increased lipid peroxidation. Concentrations of thiobarbituric acid-reactive substances correlated negatively with both plasma retinol and plasma tocopherol (r2 = .42, p < .01 and r2 = .48, p < .005, respectively). In the five patients from whom we were able to collect urine, nitrite excretion was increased approximately 400-fold (p < .001). CONCLUSIONS: These data indicate decreased antioxidant status in the face of enhanced free radical activity, and suggest potential therapeutic strategies involving antioxidant repletion.

Nitric oxide synthase activity is increased in patients with sepsis syndrome.

1. We measured nitric oxide synthase activity in peripheral blood polymorphonuclear leucocytes from 10 patients with sepsis syndrome and 10 healthy subjects. 2. Synthase activity was significantly higher in patients with sepsis than in control subjects (1202 +/- 579 compared with 595 +/- 544 pmol of nitric oxide min-1 mg-1 of cell protein, P < 0.05). 3. Activity was greatest in those patients with the larger number of organ failures, although this failed to reach significance (1489 +/- 560 in patients with three or more organ failures and 843 +/- 404 pmol of nitric oxide min-1 mg-1 of cell protein in those with less than three, P = 0.11). 4. This study provides evidence for the role of overproduction of the vasodilator nitric oxide in sepsis syndrome.

Nitric oxide production by human peripheral blood polymorphonuclear leucocytes.

1. We describe a rapid and reliable technique for the assessment of basal nitric oxide release in clinical situations, using peripheral blood polymorphonuclear leucocytes isolated by a single-step density gradient procedure. The assay is based on the quantitative conversion of oxyhaemoglobin to methaemoglobin by nitric oxide. We have further examined the ability of these cells to respond to various stimuli. 2. Basal (unstimulated) nitric oxide release occurred, which was augmented by superoxide dismutase. The mean value for healthy subjects was 283 +/- 96.7 pmol min-1 10(-6) cells. 3. Both phorbol myristate acetate and N-formyl-methionyl-leucylphenylalanine induced further release of nitric oxide, which was increased by preincubation with lipopolysaccharide, interleukin-6 and interferon-gamma. 4. Preincubation of cells with NG-monomethyl-L-arginine or L-canavanine sulphate inhibited nitric oxide production. 5. The procedure provides a valuable tool for monitoring nitric oxide up-regulation in clinical situations.

Reperfusion injury, antioxidants and hemodynamics during orthotopic liver transplantation.

Endothelial injury occurs as a result of oxygen free radical production after ischemia and reperfusion of transplanted livers, causing hemodynamic disturbance. Patients with chronic liver disease generally have low levels of fat-soluble vitamins, which have important antioxidant roles. We therefore assessed circulating levels of the antioxidants vitamin A, vitamin E, beta-carotene and lycopene, indices of lipid peroxidation and hemodynamic changes during elective orthotopic liver transplantation in 12 patients. We found that initial antioxidant levels were severely depleted compared with healthy subjects, and in some patients carotene and lycopene levels were undetectable. Increased lipid peroxidation was also evident, as shown by thiobarbituric acid-reactive substances. On reperfusion of the liver graft, vitamin A and E levels fell (p < 0.01) and were associated with decreases in systemic vascular resistance (p < 0.02). These data show that patients undergoing liver transplant have lowered antioxidant defenses and evidence of free radical damage, which compound the additional insult of reperfusion injury. Antioxidant therapy in these patients before transplantation may ameliorate the effects of reperfusion.

Potentiation of triamcinolone-induced cleft palate in mice by maternal high dietary fat.

This study investigated whether the current range in dietary fat levels, which has arisen partly in response to some major health concerns, would affect frequency of congenital anomalies if continued into the period of early pregnancy. The effect of 5.6%, or 48% of calories from fat in the maternal diet, was tested on pregnant strain CD-1 mice injected with triamcinolone in doses of 0.01 mg, 0.02 mg, 0.04 mg, or 0.06 mg per day on days 11 through 14 of gestation. Frequency of cleft palate increased with increasing doses of triamcinolone, with clefts of the palate being rare at the two lower doses. No clefts appeared without triamcinolone on either diet. In combination with triamcinolone treatment, 226 fetuses exposed to a maternal low fat diet had normal palates and 86 had cleft palates. With exposure to high fat, 186 fetuses had normal palates and 101 had cleft palates, which was a significant increase in clefting (p < 0.05). Also, the latter group showed a greater degree of retardation in palate development (p < 0.05). Thus both a greater frequency and a more severe form of clefting support the conclusion that high dietary fat potentiated the cleft palate-producing effects of triamcinolone in mice.

Pituitary tumors in mice exposed prenatally to diethylstilbestrol.

Hyperprolactinemia and prolactinomas are among the abnormalities reported for women exposed prenatally to diethylstilbestrol (DES). To pursue this issue in an animal model replicating the other abnormalities of prenatal DES exposure, pituitary glands were studied in the offspring of CD-1 mice receiving an i.p. injection of 1 or 2 micrograms DES/g body weight during late pregnancy. Among 132 mice exposed prenatally to DES and then raised to terminal illness, there were 24 pituitary tumors compared to only 1 tumor among 64 controls. The tumors consisted predominantly of cells with an eccentric nucleus and cytoplasm characterized by an acidophilic core and basophilic rim. These cells were identified as lactotrophs on the basis of prolactin immunohistochemistry and by an expected variation in frequency relative to physiological states. Evaluation of ovaries from the same mice revealed a deficiency of corpora lutea and an elevated incidence of ovarian tumors. These findings are consistent with abnormal sex differentiation of the fetal hypothalamus being the cause of most adverse effects from prenatal DES exposure.