2-Deoxy-D-glucose fails to induce feeding in hamsters fed a preferred diet

Hamsters fed a lab chow diet have been found not to increase their food intake when injected with 2-deoxy-D-glucose (2-DG). Lab chow is not a preferred diet for hamsters, however, and rats also do not eat in response to 2-DG when fed unpalatable foods. The present experiment therefore investigated the feeding response to 2-DG of adult male hamsters (Mesocricetus auratus) fed a preferred sunflower seed diet. The failure of the hamsters to increase their intake of sunflower seeds, as well as of lab chow, following injections of 750 or 1000 mg/kg of 2-DG supports the conclusion that hamsters lack a glucoprivic feeding system.     

Distinct Leishmania braziliensis isolates induce different paces of chemokine expression patterns

Inflammatory events during Leishmania braziliensis infection in mice were investigated. Large lesions were directly correlated with the inflammatory reaction but not with parasite burden. Different L. braziliensis strains induce different paces of chemokine expression patterns, leading to diverse cell recruitment and differential inflammatory responses.     

The relationship of spontaneous macronutrient and sodium intake with fluid ingestion and thirst in humans

The amount of solid food eaten by humans in spontaneously ingested bouts is the most important determinant of the amount and timing of fluid ingestion. In order to investigate whether this relationship occurred as a result of the osmotic and volumetric effects of the ingested nutrients, analyses were performed on the data obtained from 219 adult humans. They were paid to maintain diaries for 7 days of everything they ingested, the timing and conditions present at the bout, and pre- and postbout self-ratings of subjective thirst. Carbohydrate and protein intake were found to be the dietary constituents that were most highly related to fluid intake and subjective thirst while sodium and fat were found to be either not at all or only weakly related. Carbohydrate and protein intake were found to be positively related to the amount ingested of total fluid, fluid in excess of digestive requirements, and fluid in the form of "drinks," for the amounts ingested in individual bouts, over the entire day, and over the entire week. In addition, carbohydrate and protein intakes were found to be positively related to the reduction in the subjective state of thirst, while negatively related to the level of thirst self-reported at the end of the bout. The results indicate that fluid intake and subjective thirst are influenced by the repleting characteristics of ingested nutrients and not by their depleting effects, suggesting that fluid intake occurs in response to and as an adjunct of food intake, not fluid homeostasis.     

Effects of 5-Thio-D-glucose and 2-Deoxy-D-glucose upon stomach-emptying in the golden hamster

5-Thio-D-glucose (200 and 500 mg/kg) produced hyperglycemia and significantly retarded emptying of the pregastric pouch of hamsters. 2-Deoxy-D-glucose did not affect stomach-emptying at either of the doses used (500 and 1000 mg/kg), but did cause hyperglycemia at the higher dose. These results are discussed in relation to the failure of these glucose analogs to produce hyperphagia in the golden hamster.     

Feeding and macronutrient selection patterns in rats: adrenalectomy and chronic corticosterone replacement

To analyze further the role of corticosterone (CORT) in the control of feeding behavior, we examined the impact of adrenalectomy (ADX) and chronic CORT implants on the food intake and macronutrient self-selection patterns of adult male rats at different periods of the diurnal cycle. Consistent with a separate study of acute CORT injection in ADX rats (Kumar and Leibowitz, 1988), the present findings indicate that ADX significantly attenuated the rats' daily (24 hr) ingestion of all three macronutrients, namely, protein, carbohydrate and fat. However, food intake in the dark cycle, specifically during the first few hours after dark onset, was significantly more affected (-70%) than feeding in the later dark and light periods (-25%). Moreover, during this early dark time when circulating CORT level normally peaks, ADX appeared to have its strongest suppressive effect on carbohydrate ingestion. Chronic subcutaneous CORT implants in the ADX animals reversed these effects of surgery and generally restored the rats' eating patterns to that of the cholesterol-implanted SHAM animals. These findings suggest that CORT exerts a decisive influence on caloric intake, on the diurnal pattern of feeding, and on appetite for specific macronutrients. The impact of CORT on carbohydrate intake is apparent specifically during the active eating period, particularly at dark onset when endogenous CORT levels normally peak and carbohydrate is exhibited as the preferred macronutrient.     

Effects of 2-deoxy-D-glucose on schedule dependent and schedule induced behavior and ingestion at different body weights.

The effects of intraperitoneally administered 2-deoxy-D-glucose (2DG) at 0, 50, 150, 300 and 600 mg/kg, were studied in four experiments. Experiments 1 and 2 determined the effects of 2DG on schedule induced licking and drinking and schedule dependent lever pressing in rats maintained at 80% and ad lib free feeding body weights during 3 hr on an FI-1 min food reinforcement schedule. Experiments 3 and 4 evaluated the effects of 2DG on free access home cage food and water consumption under the same body weight conditions. Decreases in food and water consumption were observed under both feeding conditions when animals were reduced to 80% free feeding body weight. At ad lib feeding body weight, 2DG administration had no effects on licking, drinking, and lever pressing during FI-1 min scheduled food reinforcement or on free access home cage feeding and drinking. The effects of 2DG on ingestive behavior are discussed in terms of central and peripheral glucosensitive cells and the consequences of blocking glucose utilization at different body weights under different stimulus conditions.     

Developmental patterns of macronutrient intake in female and male rats from weaning to maturity

Male and female Sprague-Dawley rats had available pure macronutrient diets, protein, carbohydrate and fat, from birth to day 77 of age. Analyses of their intake of these nitrients, as a function of age, demonstrate that, in both sexes, daily protein intake and preference for this nutrient relative to the other macronutrients rise steadily from weaning and peak precisely at the time of puberty (day 37–44 for females and days 42–49 for males), when there is also a peak in body weight gain. This is in contrast to daily carbohydrate intake, which peaks 2 weeks after puberty in males, and also to the female and male rats' preference for carbohydrate, which remains relatively stable from weaning to maturity. These patterns also differ from those observed for daily fat intake and fat preference for females and males, which are relatively high during the first postweaning week and then decline and remain very low until shortly after puberty (day 54), when there occurs a sharp burst in fat intake. Comparisons between the females and males reveal a significantly stronger preference for carbohydrate in the females, exhibited as early as 23 days of age; a stronger preference for protein and fat in the males, evident after day 28; and greater light-period feeding of carbohydrate and fat by females compared to males, apparent after puberty. Correlational analyses demonstrate that body weight and total kcal intake are closely related to daily protein consumption, more strongly in females compared to males; are strongly related to daily fat intake only in males; and are unrelated to intake of carbohydrate, at any age and in either sex.     

The effect of caffeine ingestion on the perceived instability of visual patterns.

The unique cytoarchitecture of the visual cortex, with cells of particular orientation-specificity arranged in vertical columns adjacent to other columns with somewhat different orientation-specificity causes subjective visual instabilities when viewing some repetitive grid patterns for normal subjects. Since caffeine increases cortical arousal by serving as an antagonist to the inhibitory neurotransmitter adenosine, caffeine intake might be expected to increase these subjective disturbances. Twenty subjects were administered a placebo, 100 or 200 mg of caffeine orally. After 40 min subjects rated the level of visual instability on nine subjective dimensions, while viewing grating patterns. Each subject was tested on all levels of caffeine intake, with at least 48 h between tests. With increasing caffeine dosage the level of reported visual disturbances increased. These data suggest that the increased cortical arousal associated with caffeine intake may interact with the structural properties of the visual cortex to increase the perceptual instability associated with viewing grid patterns.     

2-Deoxy-D-glucose inhibits intracellular multiplication and promotes intracellular killing of Legionella pneumophila in A/J mouse macrophages.

Legionella pneumophila can grow intracellularly in A/J mouse macrophages. 2-Deoxy-D-glucose (2dG) (0.1, 1, and 10 mM) inhibited intracellular multiplication and promoted intracellular killing of L. pneumophila dose dependently when it was added to the culture medium of macrophage monolayers, whereas it did not inhibit the bacterial growth in buffered yeast extract broth, which was used for an L. pneumophila culture. The effect of 2dG was reversible because the surviving bacteria resumed intracellular multiplication after the washing away of 2dG from the culture. The effect of 2dG was also competitively inhibited by high concentrations of glucose. The inhibitory effect of 2dG was not attributed to the inhibition of bacterial phagocytosis by macrophages. Furthermore, sodium fluoride (0.1 and 1 mM), cycloheximide (0.1 and 1 microgram/ml), and tunicamycin (1, 2, and 5 micrograms/ml) did not promote the killing of L. pneumophila in macrophages, implying that the inhibitory effect of 2dG cannot be attributed to the inhibition of glycolysis, protein synthesis, and protein glycosylation in macrophages. We suggest that 2dG promotes intracellular killing of L. pneumophila by activating some novel killing mechanism of macrophages.     

Haemophilus influenzae and Streptococcus pneumoniae induce different intracerebral mRNA cytokine patterns during the course of experimental bacterial meningitis

Using in situ hybridization with radiolabelled oligonucleotide probes, we studied the mRNA expression of IL-1β, IL-4, IL-6, IL-10, IL-12, tumour necrosis factor-alpha (TNF-α), TNF-β, interferon-gamma (IFN-γ), and transforming growth factor-beta (TGF-β) in the brain during the lethal course of experimental meningitis in a rat model inoculated intracisternally with Haemophilus influenzae type b (Hib) or Streptococcus pneumoniae and in uninfected control rats inoculated with the same volume of PBS. The production of IL-1β, IL-4, IL-6 and IFN-γ was also evaluated by immunohistochemistry. In the brain of Hib-inoculated rats, there was marked mRNA expression of IL-1β, IL-6, TNF-α, IL-12 and IFN-γ. IL-1β, IL-6 and TNF-α were up-regulated throughout the observation period at 2, 8 and 18 h post-inoculation (p.i.), with similar patterns of induction. The Th1 cytokines IFN-γ and TNF-β were up-regulated within 8 h p.i. IL-10 and TGF-β were down-regulated at 18 h p.i., while IL-4 was not detected. In contrast, the brain of S. pneumoniae-inoculated rats showed lower levels of IL-1β, IL-6 and TNF-α, but higher levels of TNF-β and detectable mRNA expression of IL-4 when compared with Hib-inoculated rats. IL-12, IFN-γ, IL-10 and TGF-β exhibited similar patterns of induction in the brains of Hib- and S. pneumoniae-inoculated rats. At 18 h p.i., immunohistochemistry showed similar patterns of IL-1β, IL-4, IL-6 and IFN-γ as mRNA expression in the brains of Hib- and S. pneumoniae-inoculated rats. The differences of cytokine profiles induced by the two bacterial strains may imply that different immunomodulating approaches should be considered, depending on etiology.     

Intermediate- and high-affinity interleukin-2 receptors expressed in an IL-4-dependent T-cell line induce different signals.

We have previously described a synergism between the two physiological hormones, retinoic acid (RA) and 1 alpha,25-dihydroxyvitamin D3 (VD) in the induction of U937 cell differentiation towards a more mature state. Herein, we investigated the regulation of cytokine production during RA and/or VD treatment of U937 cells. Cell differentiation was followed by measurement of their capacity to give oxidative responses, and interleukin-1 beta (IL-1 beta), tumour necrosis factor-alpha (TNF-alpha) and IL-6 gene and protein expression were determined in RA/VD-treated cells, activated or not with lipopolysaccharide (LPS). The undifferentiated and RA-treated U937 cells were unable to produce monokines even when they were stimulated by LPS. VD induced the monokine mRNA expression in U937 cells but failed to induce protein release. However, unlike RA, it primed the cells to secrete monokines upon endotoxin stimulation. A large enhancement of the production of the monokines both at mRNA and protein levels was observed in the U937 cells exposed to the combination of RA + VD. Nevertheless, protein release required a further step of activation of the RA + VD-primed cells. The co-inducer effect of RA and VD was not observed in HL-60 or THP-1 cells and seems to be restricted to U937 cells. These results on cytokine expression support our previous finding that a combination of RA and VD brings the U937 cells to a high stage of myeloid differentiation with major characteristics of monocytes/macrophages.     

Purified recombinant A. fumigatus allergens induce different responses in mice

Aspergillus fumigatus an opportunistic fungus is associated with a number of diseases in humans. Allergy resulting from exposure to the A. fumigatus allergens has been recognized frequently. The damage caused by the disease is very striking in patients with atopy and those with cystic fibrosis. Avoidance to exposure is not feasible because A. fumigatus spores are ubiquitously distributed in the environment. Hence, immunotherapeutic regimens in severe forms of A. fumigatus allergy may have a high potential. However, before such forms of therapy can be envisaged, it is essential to understand the immunopathogenesis. In the present study, we investigated the role of purified A. fumigatus allergens in the development of allergic asthma in mice. We have used four major recombinant A. fumigatus allergens in the murine model. Mice exposed to Asp f 1, f 3, and f 4 showed inflammatory changes in the lungs and airway hyperreactivity. The immune responses, including elevated serum IgE, enhanced eosinophils, recruitment in the peripheral blood and lungs, and expression of regulatory cytokines, are characteristic of a Th2 response. Asp f 6 demonstrated only a reduced response in these animals. The results suggest that the pathology induced by crude A. fumigatus extract results from the cumulative effects of the allergens and the individual responses varied considerably with different purified antigens.     

Cholera toxin and Salmonella typhimurium induce different cytokine profiles in the gastrointestinal tract.

Salmonella infection of the gastrointestinal tract (GT) results in fluid secretion and inflammation. In contrast, cholera toxin (CT) induces fluid secretion but no inflammation. Using a murine ligated intestinal loop model, we investigated cytokine production (interleukin-1 [IL-1], IL-2, IL-4, IL-6, IL-10, gamma interferon, and tumor necrosis factor alpha) in the GT following exposure to these agents. Salmonella typhimurium induced a Th1-like cytokine profile in loops obtained from either nonimmune mice or Salmonella-immunized mice. CT induced only IL-6 and IL-10 production in ligated loops from nonimmune mice but induced a Th2-like cytokine profile in ligated loops obtained from CT-immunized mice. These results show that CT and S. typhimurium induce very different cytokine profiles in the GT.     

Effects of adrenal steroid agonists on food intake and macronutrient selection.

These experiments tested the effects of subcutaneous (SC) and paraventricular nucleus (PVN) administration of the steroid receptor agonists, corticosterone (CORT), aldosterone (ALDO), RU28362, and dexamethasone (DEX), on food intake and macronutrient selection during the first h of the dark feeding period in the rat. Results indicate that CORT and the selective type II receptor agonist RU28362 specifically stimulate carbohydrate ingestion after SC or PVN administration, while DEX has no effect on feeding. This selective effect of SC CORT on carbohydrate ingestion is dose dependent, seen at doses ranging from 0.125 to 2.0 mg/kg. Moreover, the stimulatory effects of CORT and RU28362 on carbohydrate intake are observed in ADX rats but not in sham rats. This is in contrast to SC and PVN administration of the type I receptor agonist ALDO, which specifically enhances fat ingestion in both sham and ADX rats. These results, with both peripheral and central steroid administration, reveal selective effects of type I and type II receptor stimulation on fat and carbohydrate intake, respectively.     

Tolerance and suppression of immunity to herpes simplex virus: different presentations of antigens induce different types of suppressor cells.

In this report, we examine tolerance (hyporesponsiveness) and suppression of delayed hypersensitivity (DH) to herpes simplex virus (HSV) in mice, using two different forms of tolerogen: HSV particles and HSV-infected spleen cells. The intravenous injection of mice with either HSV particles or spleen cells 7 days before subcutaneous immunization with virus induced a profound state of unresponsiveness. This unresponsive state was mediated, at least in part, by suppressor T cells (Ts), which were demonstrated by passive transfer to naive recipients. However, different types of Ts were induced depending on the form of the tolerogen. The injection of HSV particles induced Ts which suppressed the induction but not the expression of DH. On the other hand, the injection of HSV spleen cells induced two types of Ts: one which inhibited the induction of the DH response and one which inhibited the expression of DH to HSV. Both tolerance and Ts are virus specific (i.e., the DH response to an unrelated virus was not inhibited) but not type specific for HSV type 1 and HSV type 2. Since both virus particles and virus-infected cells may be present in the blood during HSV infection, the induction of this type of immune regulation may influence the outcome of both acute and latent HSV infections.     

Phagocytosis. Clinical disorders of recognition and ingestion.

Tentative conclusions concerning the role of recognition and ingestion of microorganisms by phagocytes in host defense and the consequences of disorders of phagocytosis can be derived by correlating a) knowledge about recognition and ingestion derived from studies in vitro, b) investigations of the clearance of particulate matter from the circulation of animals and man, and c) analyses of the behavior of phagocytes in patients susceptible to recurrent pyogenic infections. Deficiency of the major serum recognition-conferring (immunoglobulins and complement proteins that deposit a fragment of C3 on microbes) prevents the optimal clearance of virulent encapsulated pathogens by fixed mononuclear phagocytes. Confrontation of phagocytes with particulate matter appearing in pathologic states (viruses, immune complexes, damaged erythrocytes in sickle cell anemia and other hemoglobinopathies) diverts them from their normal task of clearing opsonized encapsulated microorganisms. Corticosteroids impair the phagocytic capacity by an unknown mechanism. Major impediments to progress in this field are inadequate assays for phagocytosis and the difficulty in measuring phagocytosis in the intact organism.     

Liver-derived cell lines QSG-7701 and HepG2 support different HBV replication patterns

Summary. Hepatitis B virus (HBV) infection is currently still a worldwide heath concern. In our study, we compared HBV replication patterns in two liver-derived cell lines, QSG-7701 and HepG2. Viral markers of HBV replication in culture medium and cells were analyzed after transfection of these cells with plasmid pUC18-HBV1.2 into. We showed that QSG-7701 cells could support more stable and a higher level of HBV replication than HepG2 cells. Gene expression profiles of QSG-7701 and HepG2 cells by microarray analysis showed that many genes were differentially expressed between these two cell lines, including those that are related to the HBV life cycle. The global gene expression profile of these two cell types provides some clues to explain how different HBV replication is achieved. QSG-7701 cells offer a new opportunity for basic research on HBV virus-host interactions.     

Superantigens and conventional antigens induce different responses in alpha beta T-cell receptor transgenic mice.

While superantigens such as staphylococcal enterotoxin B (SEB) have been shown to induce both clonal deletion and clonal anergy, it is still not known why tolerance rather than memory is induced. To address this issue, we tested the proliferative capacity of T cells from ovalbumin (OVA)-specific alpha beta T-cell receptor transgenic mice primed with either SEB emulsified in complete Freund's adjuvant (CFA) or with OVA peptide, the specific antigen, in CFA. By contrast cells from mice primed with SEB in CFA appeared to be anergic in that they were hyporesponsive to OVA peptide as well as to SEB. The anergic cells could respond to phorbol myristate acetate (PMA) and ionomycin, suggesting that a proximal signal transduction step was affected. Cells from transgenic mice primed with OVA peptide and CFA were not anergic and in fact displayed an enhanced response when they were challenged with OVA in vitro. Thus, when the two antigens are emulsified in CFA and then injected subcutaneously, they behave very differently: the superantigen SEB induces anergy whereas the conventional antigen OVA induces a memory type of response.