Aneuploidy assessed by DNA index influences the effect of iron status on plasma and/or supernatant cytokine levels and progression of cells through the cell cycle in a mouse model.

Aneuploidy, a condition associated with altered chromosome number, hence DNA index, is frequently seen in many diseases including cancers and affects immunity. Iron, an essential nutrient for humans, modulates the immune function and the proliferation of normal and cancer cells. To determine whether impaired immunity seen in iron-deficient subjects may be related to aneuploidy, we measured spleen cell DNA index, percent of cells in different phases of the cell cycle, plasma and/or supernatant IL-2, IL-10, IL-12, and interferon-gamma in control, pair-fed, iron-deficient, and iron-replete mice (N=20-22/group). The test and control diets differed only in iron content (0.09mmol/kg versus 0.9mmol/kg) and were fed for 68days. Mean levels of hemoglobin and liver iron stores of iron-deficient and iron-replete mice were 40-60% lower than those of control and pair-fed mice (P<0.05). Mean plasma levels of IL-10, interferon-gamma and percent of cells in S+G2/M phases were lower in mice with than in those without aneuploidy (P<0.05). Lowest plasma IL-12 and interferon-gamma concentrations were observed in iron-deficient mice with aneuploidy. Mean percents of cultures with aneuploidy and DNA indexes were higher in iron-deficient and iron-replete than in control and pair-fed mice likely due to delayed cell division (P<0.05). Aneuploidy decreased the concentration of IL-2 and interferon-gamma in baseline cultures while it increased that of interferon-gamma in anti-CD3 treated cultures. Aneuploidic indexes negatively correlated with cytokine levels, percents of cells in S+G2/M phases and indicators of iron status (P<0.05). Although chromosome cytogenetics was not performed, for the first time, we report that increased aneuploidy rate may modulate the immune function during iron-deficiency.

Ceruloplasmin and Alpha-1-Acid Glycoprotein, but not C-Reactive Protein, Correlate With Serum Ferritin During Various Postpartum/Lactation Periods in Congolese Females.

Background: Serum ferritin usually correlates positively with acute phase proteins (APPs), but limited information is available on this association during various postpartum/lactation periods. The objective of this study was to assess the association between serum ferritin and APPs in Congolese females during different postpartum/lactation periods. Methods: Serum ferritin, C-reactive protein (CRP), alpha-1-acid glycoprotein (AGP), ceruloplasmin (Cp), and transferrin saturation (TS) were measured during various postpartum/lactation periods (0.5 to 6, 6.1 to 12, 12.1 to 18, and 18.1 to 24 months) in 131 Congolese females aged 15 to 45 years. Results: Mean serum ferritin concentrations were lower in females in the 0.5- to 6-month postpartum/lactation subgroup than in the other 3 subgroups (P<0.05). Mean concentrations of hemoglobin, APPs, and TS were not different among the 4 subgroups. While serum ferritin concentrations correlated with Cp (r=0.514) and AGP (r=0.795) during the 0.5- to 6-month and the 6.1- to 12-month postpartum/lactation periods, respectively (P<0.05), they did not correlate with CRP. Multiple regression analysis suggested that Cp explained 25% of serum ferritin variance in the 0.5- to 6-month postpartum/lactation period (39.3% at 0.5 to 4 months) and AGP explained 60.5% of the variance in the 6.1- to 12-month period (3.7% at 0.5 to 4 months). CRP explained <5% of the serum ferritin variance at these postpartum/lactation periods. APPs explained ≤15.1% of serum ferritin variance at postpartum/lactation periods >12 months. Conclusion: Data suggest that the association between serum ferritin and inflammation is dependent on APP type and lactation time. This association may affect the diagnosis of iron deficiency in lactating females. The positive association between serum ferritin and Cp at 0.5 to 6 months postpartum may be necessary to increase liver iron release and erythropoiesis after childbirth.

White button and shiitake mushrooms reduce the incidence and severity of collagen-induced arthritis in dilute brown non-agouti mice.

Exotic mushrooms have been used in ancient Chinese medicine due to their immunomodulatory properties for the treatment and/or prevention of chronic diseases. However, only limited data exist on the health benefits of white button mushrooms (WBM), the most common in the American diet. In the current study, we investigated the effects of WBM and shiitake mushrooms (SM) on collagen-induced arthritis (CIA) using a 2 x 3 factorial design in 8-wk-old female dilute brown non-agouti mice that were fed a control diet (n = 37) or the same diet supplemented with 5% lyophilized WBM or SM (n = 27) for 6 wk. CIA was induced by immunizing mice with 100 µg bovine collagen followed by 50 µg LPS on d 20 post-collagen injection. CIA was assessed by mononuclear cell infiltration, bone erosion, plasma IL-6, TNFα, and intercellular adhesion molecule-1 (sICAM-1) concentrations. Compared with the control diet, WBM and SM tended to reduce the CIA index from 5.11 ± 0.82 to 3.15 ± 0.95 (P = 0.06) (median, 6-9 to 1-2) 31 d post-collagen injection. Whereas 58% of control mice had a CIA index ≥ 7, only 23% of WBM and 29% of SM mice did (P = 0.1). Although both types of mushrooms reduced plasma TNFα (34%, WBM; 64%, SM), only SM increased plasma IL-6 by 1.3-fold (P < 0.05). The CIA index was positively correlated with sICAM1 (r = 0.55; P < 0.05) but negatively correlated with TNFα (r = 0.34; P < 0.05). Whether mushrooms are beneficial for arthritis management remains to be investigated. To our knowledge, this is the first report demonstrating a possible health benefit of WBM in arthritis treatment.

Mango modulates body fat and plasma glucose and lipids in mice fed a high-fat diet.

Consumption of fruits and vegetables has been investigated for their role in the prevention of many chronic conditions. Among the fruits, mango provides numerous bioactive compounds such as carotenoids, vitamin C and phenolic compounds, which have been shown to have antioxidant and anti-inflammatory properties. The present study examined the effects of dietary supplementation of freeze-dried mango pulp, in comparison with the hypolipidaemic drug, fenofibrate, and the hypoglycaemic drug, rosiglitazone, in reducing adiposity and alterations in glucose metabolism and lipid profile in mice fed a high-fat (HF) diet. Male C57BL/6J mice were randomly divided into six treatment groups (eight to nine/group): control (10 % energy from fat); HF (60 % energy from fat); HF+1 or 10 % freeze-dried mango (w/w); HF+fenofibrate (500 mg/kg diet); HF+rosiglitazone (50 mg/kg diet). After 8 weeks of treatment, mice receiving the HF diet had a higher percentage body fat (P = 0·0205) and epididymal fat mass (P = 0·0037) compared with the other treatment groups. Both doses of freeze-dried mango, similar to fenofibrate and rosiglitazone, prevented the increase in epididymal fat mass and the percentage of body fat. Freeze-dried mango supplementation at the 1 % dose improved glucose tolerance as shown by approximately 35 % lower blood glucose area under the curve compared with the HF group. Moreover, freeze-dried mango lowered insulin resistance, as indicated by the homeostasis model assessment of insulin resistance, to a similar extent as rosiglitazone and modulated NEFA. The present findings demonstrate that incorporation of freeze-dried mango in the diet of mice improved glucose tolerance and lipid profile and reduced adiposity associated with a HF diet.

Iron deficiency, but not underfeeding reduces the secretion of interferon-gamma by mitogen-activated murine spleen cells.

Interferon-gamma (IFN-γ), a cytokine primarily secreted by T and natural killer cells regulates cell-mediated and innate immunity. Iron deficiency, a public health problem in children impairs immune function. To determine whether reduced IFN-γ contributes to impaired immunity, we measured IFN-γ in supernatants of activated (2.5 µg/ml concanavalin A, 50 ng/ml anti-CD3 antibody) spleen cells from control (C), iron-deficient (ID), pair-fed (PF), and iron-replete mice for 3 (R3) and 14 days (R14) (11-12/group). Except for iron content, the low iron (5 ppm) and control (50 ppm) diets had identical composition. Mean indices of iron status after 51 days of feeding were as follows: C=PF≈R14>R3>ID (p<0.01). Iron deficiency, but not pairfeeding reduced IFN-γ concentration in mitogen-treated cells by 30-43% (p<0.05); iron repletion improved it. Reduced IFN-γ was not simply due to differences in IL-12 (IFN-γ inducer), percentage of CD3+ T cells, or impaired cell proliferation because these indices were not always decreased. It was likely due to a defect in T cell activation that leads to IFN-γ gene expression. IFN-γ positively correlated with indicators of iron status, body, and thymus weights (r=0.238-0.472; p<0.05). Reduced IFN-γ secretion during iron deficiency may affect response to infections.

In Vitro Lymphocyte Functions in Undernourished Children With Sickle Cell Anemia.

Background: Children with sickle cell disease (SCD) often suffer from growth deficits and impaired immunity. However, the association between mild to moderate malnutrition and in vitro lymphocyte function has not been well studied. The goal of this study was to investigate the effects of undernutrition on lymphocyte functions in children with SCD. Methods: Weight; height; plasma concentrations of albumin (Alb), prealbumin (PA), transferrin (Tf), retinol-binding protein (RBP), α1-acid glycoprotein (AGP), C-reactive protein (CRP), and ceruloplasmin (Cp); and lymphocyte proliferation and interleukin (IL)-2 in phytohemagglutinin-treated blood lymphocytes were measured in 90 children with SCD (59 SS, 4 Sß°, 27 SC hemoglobin genotypes). Results: The mean age of the children included in the analysis was 7.65 years. Four of the 90 children had weight and height below the fifth percentile. A higher percentage of children with HbSS/HbSß° (61.4%) than of those with HbSC (44%) had ≥2 plasma protein concentrations below normal (Alb <35 g/L, PA <160 mg/L, Tf <2.0 g/L, and RBP ≤20 mg/L). Mean anthropometric measurements, hemoglobin, and hematocrit were lower in children with HbSS/HbSß° than in those with HbSC (P<0.05). Lymphocyte proliferation was reduced by 20% to 27% in children with HbSS/HbSß° with undernutrition plus inflammation (AGP >1 g/L, CRP >5 mg/L, Cp >600 mg/L) compared to children with neither. Regardless of inflammatory status, lymphocyte proliferation was reduced by 29% to 49% in children with HbSS/HbSß° and undernutrition defined by PA or Alb plus RBP (P<0.05) compared to those with RBP within normal range. Neither undernutrition nor inflammation reduced lymphocyte proliferation in children with HbSC. Mean IL-2 activity was reduced by undernutrition, defined as PA <160 mg/L, in both groups. PA, RBP, and hemoglobin concentrations positively correlated with in vitro lymphocyte functions (P<0.05). Conclusion: Undernutrition altered in vitro lymphocyte function in children with the HbSS/HbSß° genotypes. Dietary supplements may improve the altered functions in these children.

Unusual low plasma levels of zinc in non-pregnant Congolese women.

Zn is an essential trace element required throughout the life cycle. Although suboptimal Zn status is thought to be common in many sub-Saharan countries, there is a paucity of data in the Democratic Republic of Congo. The objective of the study was to determine Zn status in non-pregnant Congolese women. We measured plasma Zn and indicators of nutritional status (albumin, prealbumin, retinol-binding protein) and inflammation (C-reactive protein (CRP), ceruloplasmin, alpha1-acid glycoprotein (AGP)) in seventy-seven lactating and thirty non-lactating women (mean age 28 and 31 years, respectively). Blood samples were collected in summer 1989 in rural Bas-Congo during a survey on Fe status. Mean lactation period was 8.3 months. Mean parity was higher in lactating (3.6) than in non-lactating (2.2) women (P < 0.05). Mean biochemical indicators of nutritional status, CRP and ceruloplasmin were within normal range and not different between groups. Mean AGP concentrations were above normal (>1.2 g/l) and higher in lactating (1.365 g/l) than in non-lactating (1.178 g/l) women (P < 0.05). Mean Zn concentration (540 microg/l) of the overall study population was below normal (700 microg/l); and the mean was lower in lactating (455 microg/l) than in non-lactating (759 microg/l) women (P < 0.05). Multiple regression analysis suggested that parity (P < 0.05), but not inflammation, was the most important factor associated with low Zn levels. Despite the lack of data on dietary intake, the results suggest that suboptimal Zn status may be common in the studied population.

The Inhibitory Properties of Ethanol Extracts of Some Culinary-Medicinal Mushrooms on the Secretion of Interleukin-8 and Vascular Endothelial Growth Factor by PC3 Cancer Cells.

Interleukin (IL)-8, a cytokine produced by immune and non-immune cells, induces angiogenesis via increased vascular endothelial growth factor (VEGF) secretion; both cytokines promote tumor growth. IL-8 and VEGF plasma levels correlate with prostate cancer severity, suggesting that therapeutic options aimed at their downregulation may modulate tumor growth. Available data suggest that Agaricus bisporus (white button mushroom [WBM]) extracts inhibit cancer cell proliferation through aromatase inhibition. However, the extent to which they affect IL-8 and VEGF remains to be elucidated. The aims of this study were to (1) investigate the antiproliferative properties of WBM, brown A. bisporus (portabella), and Lentinus edodes (shiitake mushroom) on PC3 cancer cells; (2) demonstrate that these properties are exerted through the regulation of both IL-8 and VEGF; and (3) determine the role of NFκB activation in the antiproliferative process of mushroom extracts. Cytokine secretion in the supernatant, NFκB activity, and cell proliferation were measured in PC3 cells incubated with 0-100 µg/mL of ethanol extracts of mushrooms. Mushroom extracts decreased IL-8 secretion and cell proliferation (P < .05), and also tended to decrease VEGF (P < .09). Decreased cell proliferation did not appear to result from cell death because trypan blue exclusion tests showed comparable cell viability among cultures. Mushroom extracts also decreased nuclear and total NFκB activity, and the ratio of nuclear to cytoplasmic activity (P < .05) suggesting altered translocation from the cytoplasm to the nucleus. Our data suggest that the three types of studied mushrooms may modulate tumor growth through inhibition of IL-8, VEGF, and NFκB pathways.

Clinical Observations, Plasma Retinol Concentrations, and In Vitro Lymphocyte Functions in Children With Sickle Cell Disease.

BACKGROUND: Children with sickle cell disease (SCD) often have infections, growth deficits, and impaired immunity, problems that also are observed in individuals with a vitamin A deficiency (plasma retinol concentration <20 µg/dL). The goal of this study was to investigate the association between vitamin A, health status, and the in vitro immune function of children with SCD. METHODS: Fifty-nine children (40 SS, 11 SC, and 8 Sßthalassemia [Sßthal] hemoglobin genotypes) 9 months to 18 years old were investigated for plasma levels of retinol, retinol binding protein, C-reactive protein, alpha-1-acid glycoprotein, lymphocyte proliferation, and interleukin (IL)-2 activity in supernatant of phytohemagglutinin-treated lymphocytes. RESULTS: The plasma retinol concentrations of children with SCD (mean 57.6 µg/dL, range 4.6-116 µg/dL) were not different from those of 21 normal individuals (mean 62 µg/dL, range 28.7-162 µg/dL). Plasma retinol concentrations did not vary by hemoglobin genotype but were lower in boys than in girls (P < 0.05) and were also lower in children with inflammation (P = 0.1). Seven children (11.9%) (6 HbSS, 1 HbSß0thal) were vitamin A-deficient, and 9 children (15.3%) had suboptimal vitamin A status (plasma retinol concentration of 20-29 µg/dL). Children with vitamin A deficiency had slightly lower height (P = 0.09) and weight mean percentiles, lymphocyte proliferative responses, and IL-2 activity (P > 0.1), but higher means of C-reactive protein (P = 0.05), pain crisis episodes and inflammation (P = 0.1), and health scores (P > 0.1) than children who were not vitamin A-deficient. Lymphocyte proliferative responses negatively correlated with health score, pain crisis episodes, and blood units received, but positively correlated with retinol binding protein (P < 0.05 to P = 0.1). CONCLUSION: Identification and correction of suboptimal vitamin A status in children with SCD may improve immunity and attenuate certain health complications associated with this disease.

Plasma zinc levels inversely correlate with vascular cell adhesion molecule-1 concentration in children with sickle cell disease.

Zinc deficiency has been implicated in impaired cell-mediated immunity of children with sickle cell disease (SCD). However, its influence on the expression of vascular cell-adhesion molecule-1 (VCAM-1) on endothelial cells, a protein involved in vasoocclusion, has not been previously investigated. We therefore measured (soluble) sVCAM-1 and zinc in 76 SCD children and 96 non-SCD children, mean age 7.73 years and 11.24 years, respectively. Although mean zinc levels of both groups were within the normal range (approximately 14.5 micromol/l), 14.5 % of SCD and 11% of non-SCD children (without inflammation) had levels below normal (10.7 micromol/L). Mean sVCAM-1 concentrations of SCD children (837 microg/l) were significantly higher than those of controls (627 microg/l) (p < 0.001). Differences persisted after taking into account age, hemoglobin phenotype, and inflammation (alpha-l acid glycoprotein >l g/l and C-reactive protein >10 mg/I). sVCAM-1 negatively correlated with serum (r = -0.444) and red blood cells zinc (r = -0.242, p < 0.05) but not with acute-phase proteins. Mean sVCAM-1 tended to be higher in SCD children with than in those without a history of a health problem (infection, pain crisis or were transfused; not significant). Data suggest that zinc may modulate the clinical status of SCD children through VCAM-1 expression, and zinc supplementation may be beneficial in these patients.

Hydroxyurea and Zileuton Differentially Modulate Cell Proliferation and Interleukin-2 Secretion by Murine Spleen Cells: Possible Implication on the Immune Function and Risk of Pain Crisis in Patients with Sickle Cell Disease.

BACKGROUND: Hydroxyurea (HU) reduces major complications associated with sickle cell disease in part because of the induction of fetal hemoglobin. However, because of its antiproliferative property, its long-term use may impair immunity. Zileuton, a derivative of HU, also induces fetal hemoglobin and has antiinflammatory properties, a feature that can reduce the risk of sickling. Our goal was to investigate the capacity of both drugs to modulate the secretion of interleukin-2 (IL-2), a regulatory cytokine for immune responses. METHODS: Spleen cells obtained from 11 4-month-old C57BL/6 female mice were incubated without and with 10 µg/mL HU or zileuton, 2.5 µg/mL concanavalin A (ConA), 20 µg/mL phytohemagglutinin (PHA), and 50 ng/mL anti-CD3 antibody for 12-48 h. IL-2 was measured in the supernatant by enzyme-linked immunosorbent assay and cell proliferation by (3)H-thymidine uptake. RESULTS: While HU reduced lymphocyte proliferation in response to mitogens (P<0.05), zileuton did not. Baseline IL-2 concentration and PHA-induced IL-2 were not significantly affected by either drug. Contrary to what we expected, while HU increased IL-2 supernatant levels 1.17-fold to 6.5-fold in anti-CD3 antibody-treated cells (P<0.05), zileuton decreased them 35%-65% (P<0.05). Zileuton likely reduced IL-2 levels by inhibiting 5-lipoxygenase, hence leukotriene B4 production, an IL-2 inducer. HU did not decrease IL-2 secretion likely because of its lack of effect on mRNA and protein synthesis. CONCLUSION: Modulation of IL-2 secretion by zileuton and/or reduced lymphocyte proliferation by HU may impair the immune response of patients with sickle cell disease but may also be beneficial by attenuating inflammation independently of fetal hemoglobin induction.

Serum ferritin levels and transferrin saturation in men with prostate cancer.

Elevated body iron stores (serum ferritin >300 microg/L, transferrin saturation TS >50%) are associated with increased risk of liver and lung cancers. To determine whether such association also exists for prostate cancer (PC), we measured serum ferritin, serum iron, total iron-binding capacity (TIBC), and TS in serum samples from 34 men with newly diagnosed, untreated PC and 84 healthy men, ranging in age from 49-78 years. In contrast with other malignancies, men with PC had significantly lower mean concentrations of serum ferritin (156 microg/L) and TS (24.35%) than those without PC (ferritin, 245 microg/L; TS, 31.98%) (p<0.05). The 95% confidence intervals for ferritin were 109-203 microg/L and 205-286 microg/L, and those for TS were 20.29-28.4% and 28.35-35.61% for men with and without PC, respectively. Significant differences were observed between both groups in the distribution of serum ferritin (<100, 101-300, >300 microg/L) and TS (<16, 16-50, >50%) (p<0.05). A lower percentage of cases than of controls had serum ferritin (17.6% versus 29.8%) and TS (5.9% versus 14.7%) above normal. These differences persisted when the analysis was limited to African-American men (31 cases and 52 controls). Data suggest that elevated body iron stores are less common in men with PC compared to those without PC.

White button, portabella, and shiitake mushroom supplementation up-regulates interleukin-23 secretion in acute dextran sodium sulfate colitis C57BL/6 mice and murine macrophage J.744.1 cell line.

Interleukin-23 (IL-23), a cytokine produced primarily by dendritic cells, is involved in host defense against gut pathogens and promotes innate immunity and inflammatory responses through the IL-23/interleukin-17 axis. We previously reported that extracts from edible mushrooms enhanced antimicrobial α-defensin production n HL60 cells. Because IL-23 is involved in defensin production, we hypothesized that edible mushrooms may modulate its secretion and gut inflammation. Eight-week-old C57BL/6 mice were fed the AIN76 diet or the same diet supplemented with 5% white button (WBM), portabella, or shiitake mushrooms. To assess in vivo and in vitro cytokine secretion, 7 to 8 mice per group received 3% dextran sodium sulfate (DSS) in drinking water during the last 5 days of the 6-week feeding period. To delineate the mechanisms by which mushrooms alter IL-23 secretion, J.744.1 cells were incubated with (100 µg/mL) WBM, portabella, and shiitake extracts without and with 100 µg/mL curdlan (a dectin-1 agonist) or 1 mg/mL laminarin (a dectin-1 antagonist). The dectin-1 receptor is a pattern-recognition receptor found in phagocytes, and its activation promotes antimicrobial innate immunity and inflammatory responses. In DSS-untreated mice, mushrooms significantly increased IL-23 plasma levels but decreased those of interleukin-6 (IL-6) (P < .05). In DSS-treated mice, mushroom-supplemented diets increased IL-6 and IL-23 levels (P < .05). Mushroom extracts potentiated curdlan-induced IL-23 secretion, and mushroom-induced IL-23 secretion was not blocked by laminarin in vitro, suggesting the involvement of both dectin-1-dependent and dectin-1-independent pathways. Although all mushrooms tended to increase IL-6 in the colon, only WBM and shiitake tended to increase IL-23 levels. These data suggest that edible mushrooms may enhance gut immunity through IL-23.

Iron deficiency reduces serum and in vitro secretion of interleukin-4 in mice independent of altered spleen cell proliferation.

Iron deficiency, a worldwide public health problem in children and adult women, impairs innate and cell-mediated immunity including interferon-γ secretion. Its effects on interleukin (IL)-4 have not been well investigated. Interleukin-4, a cytokine primarily secreted by TH2 lymphocytes, regulates B-cell proliferation and the switching of immunoglobulin (Ig)M to IgE subtypes; the latter is involved in the defense against helminth infection. Considering the fact that interferon-γ is a potent inhibitor of IL-4, we hypothesize that iron deficiency would increase the secretion of IL-4 and IgE. We measured IL-4 in serum and supernatant of concanavalin A and anti-CD3 antibody-treated spleen cells from iron-deficient, control, pair-fed DBA and C57BL/6 mice (20-24/group) and iron-replete mice for 3, 7, and 14 days (8-13/group). Feeding the low-iron diet (5 ppm vs 50 ppm for the control diet) for 2 months significantly reduced the mean levels of hemoglobin, hematocrit, liver iron stores, thymus weight, and induced splenomegaly in both strains of mice (P < .001). Iron deficiency, and not pair-feeding, reduced plasma IL-4 levels (P < .05), although it did not significantly affect IgE levels. Iron deficiency, especially when associated with thymus atrophy, reduced in vitro IL-4 secretion by activated spleen cells, cell proliferation, and percentage of CD4⁺IL-4⁺ cells (P < .05). Impaired cell proliferation did not fully explain reduced in vitro IL-4 secretion because iron-deficient mice with a normal thymus weight had a normal (3)H-thymidine uptake but decreased supernatant IL-4. It was likely due to low percentage of CD4⁺IL-4⁺. Iron repletion improved IL-4 measurements. Data suggest that iron deficiency has generalized negative effects on T-cell function. Unaltered plasma IgE may be due to other cytokines (ie, IL-13) that also modulate its secretion.

Dietary supplementation with dried plum prevents ovariectomy-induced bone loss while modulating the immune response in C57BL/6J mice.

This study was designed to investigate the effects of dried plum on the changes in bone metabolism and the immune response associated with ovarian hormone deficiency. Adult female C57BL/6J mice were either sham-operated (Sham) and fed AIN-93 diet (control) or ovariectomized (OVX) and fed a control diet with 0%, 5%, 15% or 25% dried plum (w/w), corresponding to control, low- (LDP), medium- (MDP) and high (HDP)-dose dried plum. Four weeks of HDP supplementation prevented the decrease in spine bone mineral density and content induced by OVX. The OVX compromise in trabecular bone of the vertebra and proximal tibia was prevented by the higher doses of dried plum, and in the vertebra these effects resulted in greater (P<.05) bone strength and stiffness. In the bone marrow, OVX suppressed granulocyte and committed monocyte populations and increased the lymphoblast population, but the MDP and HDP restored these myeloid and lymphoid populations to the level of the Sham. Dried plum also suppressed lymphocyte tumor necrosis factor (TNF)-α production ex vivo by splenocytes, in response to concanavalin (Con) A stimulation. These data indicate that dried plum's positive effects on bone structural and biomechanical properties coincide with the restoration of certain bone marrow myeloid and lymphoid populations, and suppressed splenocyte activation occurring with ovarian hormone deficiency.

Differential effects of shiitake- and white button mushroom-supplemented diets on hepatic steatosis in C57BL/6 mice.

Shiitake mushrooms (SMs) have been used in Asia for treatment and/or prevention of chronic diseases and hypercholesterolemia. Previously, we observed a diet supplemented with 5% SM resulted in a twofold increase in plasma IL-6 levels in DBA arthritic mice. An elevation in plasma IL-6 has also been implicated in the pathogenesis fatty liver disease. Thus, the aim of this study was to investigate the effect of SM supplemented-diet on hepatic steatosis. In study 1, eight-week old female C57BL/6 mice were randomly assigned to the following groups for 6 weeks: the AIN-93 diet; 5% SM, and 5% white button mushroom (WBM) supplemented diets (12/group). In study 2, mice were fed either the AIN-93 diet or SM (20/group). After 6 weeks, 13 mice fed SM diet were given the AIN93 diet for 8 or 15 days. Unlike other groups, all mice fed the SM diet developed fatty liver (mean histopathology score 4.5 vs <1 in the other groups; p<0.001) without fibrosis and inflammation. Fifteen days post withdrawal of SM completely normalized liver histology. To the best of our knowledge, this is the first report that chronic consumption of SM is associated with the development of fatty liver. The mechanism by which SM causes hepatic steatosis warrants further investigation.

Differential effects of isoflurane and CO2 inhalation on plasma levels of inflammatory markers associated with collagen-induced arthritis in DBA mice.

Inhalation of CO2 or isoflurane is a commonly used method of euthanasia with mice, but information related to their effects on serum inflammatory markers in chronic models of inflammation is limited. In the current study, nineteen-week old DBA female mice with (n = 53) or without (n = 51) collagen-induced arthritis were randomly assigned to euthanization with CO2 (n = 55) or isoflurane (n = 49. Plasma was collected for the measurement of soluble intercellular adhesion molecule-1 (sICAM-1), tumor necrosis factor alpha (TNF-alpha), and interleukin-6 (IL-6) by ELISA. When mice without and with collagen-induced arthritis were pooled, compared to CO2, administration of isoflurane was associated with lower production of the pro-inflammatory cytokines TNF-alpha (pg/ml, mean +/- SEM) (26.1 +/- 2.82 versus 48.1 +/- 7.99) and IL-6 (25.18 +/- 2.73 versus 48.1 +/- 6.82) (ANOVA, p < 0.05). In contrast to TNF-alpha and IL-6, administration of CO2 decreased the plasma sICAM-1 level (1170+/- 50 versus 758 +/- 24 for CO2) (p < 0.00001). When data were analyzed as a function of collagen-induced arthritis, the differences between CO2 and isoflurane persisted. Low plasma sICAM-1 levels found in CO2 euthanasia group may be due to degradation. Since mice are the most common animal model for studying inflammation, researchers should be aware of these iatrogenic experimental variables before interpreting their data.

Correlation between serum prostate-specific antigen and alpha-1-antitrypsin in men without and with prostate cancer.

Prostate specific antigen (PSA) is frequently used for prostate cancer (PCa) screening, but serum levels are also increased by prostate inflammation. Elevations in serum levels of alpha1-antitrypsin (ATT), a marker of inflammation, in cancer patients are well documented. However, an association between PSA and ATT has never been investigated. The authors, therefore, measured serum acute phase proteins (APPs) ATT, alpha1-acid glycoprotein, C-reactive protein, and alpha1-antichymotrypsin in 174 men without and 34 with newly diagnosed untreated PCa (38-80 years old). As expected, men with PCa had higher mean PSA levels than those without PCa (P < 0.00001). Men with PCa and those without PCa but with PSA >2 ng/mL (n = 68) had significantly higher ATT concentrations than those without these conditions (n = 106) (mean +/- SEM g/L): 1.94+/-0.083, 1.92+/-0.066, 1.25+/-0.043, respectively; p <0.005). Interestingly, African-American men without PCa (n=111) had higher ATT levels than Caucasian men (n=63) (1.565+/-0.045 g/l versus 1.395+/-0.056 g/l; p <0.005); and differences persisted in men with PSA >2 ng/ml (2.094+/-0.07 g/l versus 1.593 for all0.095 g/l; p<0.0002). There were no differences among groups in the levels of other APP. ATT showed the strongest correlation with PSA (r = 0.346 to 0.395; p <0.001) than any other APP (r < or =0.245). Our data suggest that men with PCa have higher ATT levels than those without PCa; and African-American men without PCa have higher ATT levels than Caucasian men. The possible implications of elevated ATT levels in African-American men on the risk of PCa are discussed.

Differential effects of hydroxyurea and zileuton on interleukin-13 secretion by activated murine spleen cells: implication on the expression of vascular cell adhesion molecule-1 and vasoocclusion in sickle cell anemia.

BACKGROUND: Interleukin-13 (IL-13), a TH2 cytokine, upregulates the expression of vascular cell adhesion molecule-1 on endothelial cells, a factor involved in vasoocclusion in sickle cell disease (SCD). Hydroxyurea improves clinical status of SCD patients in part by induction of fetal hemoglobin. Its effect on IL-13 secretion has not been investigated. OBJECTIVE: To determine whether hydroxyurea and zileuton, a hydroxyurea derivative with antiinflammatory properties, affect IL-13 secretion. METHODS: We measured IL-13 in the supernatant of murine spleen cells incubated without and with hydroxyurea, zileuton (10 microg/ml), concanavalin A (2.5 microg/ml), and anti-CD3 (50 ng/ml) (n=8). RESULTS: Hydroxyurea and zileuton do not affect baseline IL-13 secretion. Unexpectedly, hydroxyurea increases IL-13 levels above baseline (120%, 216.5%, [p<0.05] after 24 h and 48 h, respectively) in lymphocytes activated by anti-CD3, while zileuton reduces them by 59%-78% (p<0.005). In lymphocytes activated by concanavalin A, hydroxyurea and zileuton reduce IL-13 secretion by 24-36% and 50-87%, respectively (p<0.05). Hydroxyurea, but not zileuton, significantly inhibits spleen cell proliferative responses to mitogens (p<0.005). CONCLUSION: Data suggest that hydroxyurea up-regulates IL-13 secretion in anti-CD3-activated lymphocytes through gene activation but not by altered cell proliferation. Increased IL-13 secretion may contribute to unresponsiveness of certain SCD patients to hydroxyurea. The potential benefit of zileuton in the management of vasoocclusion is discussed.

Iron deficiency and in vitro iron chelation reduce the expression of cluster of differentiation molecule (CD)28 but not CD3 receptors on murine thymocytes and spleen cells.

Cluster of differentiation molecule (CD)3 and CD28 receptors play crucial roles in T-lymphocyte proliferation. Fe deficiency in man and animals impairs T-lymphocyte proliferation by unknown mechanisms. To test the hypothesis that reduced CD3 and CD28 expression is one of them, thymocytes and splenocytes from control (C; n 24), Fe-deficient (ID; n 24), pair-fed (PF; n 24), and ID mice that were Fe-repleted for 3 (R3; n 24) or 14 d (R14; n 12) were labelled with anti-CD3-fluorescein isothiocyanate and anti-CD28-phycoerythrin antibodies. Positive cells were analysed by flow cytometry. Significant differences were observed among groups in the mean levels of haemoglobin and liver Fe stores (C=PF=R14>R3>ID; P<0.005). While Fe deficiency slightly increased the percentage of CD3+ splenocytes, it reduced that of CD28+ thymocytes in mice with thymus atrophy and splenomegaly (P<0.05). These changes were corrected by Fe repletion. CD28 mean fluorescence intensity (FI) was lower and CD3 FI was higher in lymphocytes from R3 and ID, especially those with splenomegaly, than in those from R14 and PF mice (P<0.05). In vitro Fe chelation by deferoxamine (60 min) significantly decreased CD28 expression (P<0.05), and slightly increased that of CD3 (P>0.05). Spleen cell proliferative responses to concanavalin A and anti-CD3+/-anti-CD28 were reduced by Fe deficiency (ID