Purine nucleoside phosphorylase: A potential target for the development of drugs to treat T-cell- and apicomplexan parasite-mediated diseases

Purine nucleoside phosphorylase (PNP) catalyzes the reversible phosphorolysis of nucleosides and deoxynucleosides, generating ribose 1-phosphate and the purine base, which is an important step of purine catabolism pathway. The lack of such an activity in humans, owing to a genetic disorder, causes T-cell impairment, and thus drugs that inhibit human PNP activity have the potential of being utilized as modulators of the immunological system to treat leukemia, autoimmune diseases, and rejection in organ transplantation. Besides, the purine salvage pathway is the only possible way for apicomplexan parasites to obtain the building blocks for RNA and DNA synthesis, which makes PNP from these parasites an attractive target for drug development against diseases such as malaria. Hence, a number of research groups have made efforts to elucidate the mechanism of action of PNP based on structural and kinetic studies. It is conceivable that the mechanism may be different for PNPs from diverse sources, and influenced by the oligomeric state of the enzyme in solution. Furthermore, distinct transition state structures can make possible the rational design of specific inhibitors for human and apicomplexan enzymes. Here, we review the current status of these research efforts to elucidate the mechanism of PNP-catalyzed chemical reaction, focusing on the mammalian and Plamodium falciparum enzymes, targets for drug development against, respectively, T-Cell and Apicomplexan parasites-mediated diseases.

Fasting, adrenalectomy, and gluconeogenesis in the chicken and a carnivorous bird

Previous studies showed that livers from carnivorous birds have a higher gluconeogenic capacity and higher levels of gluconeogenic enzymes than livers from granivorous birds. In this work we compare the effects of fasting and adrenalectomy on gluconeogenesis. Fasting in the chicken elicited increased rates of incorporation of 14C from alanine into blood glucose, increased gluconeogenesis in liver slices, and increased activities of four gluconeogenic enzymes: glucose-6-phosphatase, phosphoenolpyruvate carboxykinase, alanine aminotransferase, and aspartate aminotransferase. These responses in the chicken resemble those observed in fasted rodents. In marked contrast, fasting in black vultures induced decreased rates of incorporation of alanine label into circulating glucose, decreased gluconeogenesis in liver slices, and no change in any of the four enzymes studied. This unusual response to fasting in the carnivorous bird is probably related to the high-protein-low-carbohydrate content of the diet. Fasted adrenalectomized birds (granivorous and carnivorous) had reduced rates of in vivo glucose synthesis, decreased liver gluconeogenesis, and lower activity of glucose-6-phosphatase and aspartate aminotransferase, without change in phosphoenolpyruvate carboxykinase and alanine aminotransferase activities.

Influence of age on the febrile response to E. coli and S. typhimurium endotoxins in growing pullets.

1. The effect of bacterial endotoxin injection was studied in growing pullets of different ages. Commercial chicks were divided into 5 groups according to age. Bacterial endotoxins (E. coli and S. typhimurium) were injected intravenously and rectal temperature was measured over a period of 300 min. 2. The results showed no significant effect of age on the febrile response induced by bacterial endotoxins, but a slight tendency towards a reduced fever peak was observed with increasing age. The response latency also increased with age.

Influence of Dietary Lysine Levels and Arginine:Lysine Ratios on Performance of Broilers Exposed to Heat or Cold Stress during the Period of Three to Six Weeks of Age

Four trials of identical experimental design were conducted to determine the effects of temperature, dietary Lys level, and dietary Arg:Lys ratios on performance and carcass yield of male broilers. Birds of a commercial strain were grown from 21 to 42 d of age in wire-floored finishing batteries placed in environmental chambers. The chambers were programmed to provide either a constant thermoneutral temperature (21.1 C), a constant cold temperature (15.5 C), or a cycling hot diurnal temperature (25.5 to 33.3 C). Within each environment there was a factorial arrangement of three Lys levels (1.0, 1.1, and 1.2%) with four Arg:Lys ratios (1.1:1, 1.2:1, 1.3:1, and 1.4:1). Environmental temperature significantly influenced virtually every characteristic examined. Hot cyclic temperatures reduced weight gain, feed intake, and breast meat yield, and increased feed conversion, dressing percentage, leg quarter yield, and abdominal fat content. The cold environment promoted increased feed intake and mortality. Ascites and cardiomyopathy were the leading causes of death under cold exposure and thermoneutral conditions, whereas complications arising from heat exposure were the main cause of death under hot cyclic conditions. Levels of Lys affected leg quarter yield and abdominal fat content over all environments but increased breast meat yield only under cold conditions. Increasing Arg: Lys ratios improved feed conversion and dressing percentage and reduced abdominal fat content; it could not be determined whether these responses were consistent with Arg per se or were due to a nonspecific N response. As increasing Lys levels or Arg:Lys ratios did not improve weight gain, increase breast meat yield, or attenuate adverse effects due to heat or cold exposure, it is concluded that the levels of Lys and Arg suggested for 21 to 42 d by the NRC are adequate for birds of this age under the environmental conditions encountered.

Metabolic Disturbances in Male Broilers of Different Strains. 1. Performance, Mortality, and Right Ventricular Hypertrophy

Two trials were carried out to test the susceptibility for metabolic disturbances of different strains of male broilers. In Trial 1, 1,890 male chickens were allotted in a randomized block design with seven treatments (Arbor Acres, Avian Farms, Cobb-500, Hubbard-Peterson, ISA, Naked Neck, and Ross) and six blocks of 45 chickens. Trial 2 involved 2,184 male chickens of six strains (Arbor Acres, Avian Farms, Cobb 500, Hubbard-Peterson, ISA Naked Neck, and Ross) allotted in seven complete blocks of 52 birds. The same management system was adopted for all birds, reared up to 42 d in an open house during late winter (Trial 1) or late autumn (Trial 2). The most marked differences observed among the strains tested was the lower BW and higher feed conversion of Naked Neck broilers. Total percentage mortalities were high among the most productive broilers, being more than 50% due to sudden death (SDS) and ascites syndrome (AS). No Naked Neck birds died as a consequence of these disturbances and the total mortalities were significantly lower (P ≤ 0.05) than the other strains. The ratio of right ventricle weight to total ventricle weight of the dead birds was over 0.25, except for Naked Neck birds, which presented a nonhypertrophic ratio. The two trials confirmed the relationship between high productivity and high incidence of SDS and AS and indicated that Naked Neck male broilers are resistant to these metabolic disturbances.

Metabolic disturbances in male broilers of different strains. 2. Relationship between the thyroid and somatotropic axes with growth rate and mortality

Seven male broiler strains (Arbor Acres, Avian Farms, Cobb-500, Hubbard-Peterson, ISA, Naked Neck, and Ross) were compared for their growth rate, feed efficiency, and mortality due to sudden death and ascites. In addition, weekly plasma levels of thyroid hormones [3,3′,5-triiodothyronine (T3) thyroxine (T4), T3: T4 ratio, growth hormone (GH), and insulin-like growth factor-I (IGF-I)] were determined. The highly productive, commercial strains were very similar in their endocrine profiles but differed markedly from the Naked Neck chickens. Naked Neck chickens were characterized by higher plasma T3 and lower T4 levels at similar ages as well as when compared on the same body weight basis. The present findings support the hypothesis that the slightly hypothyroid state of high productive broilers renders them more sensitive to metabolic disorders. Naked Neck chickens also had higher plasma GH levels than those of their age-matched commercial broilers. The coefficient of variation for GH was highest for Naked Neck chickens, which is indicative for an amplified GH burst amplitude. It may be stated that changes in plasma thyroid hormone concentration in indirect response to selection for low feed conversion and fast growth may be causatively linked to susceptibility for metabolic disturbances such as sudden death syndrome and ascites.

Sodium and chloride requirements of young broiler chickens fed corn-soybean diets (one to twenty-one days of age)

Sodium (Na+) and chloride (Cl-) nutritional requirements, dietary electrolyte balance (DEB), and their effects on acid-base balance, litter moisture, and tibial dyschondroplasia (TD) incidence for young broiler chickens were evaluated in two trials. One-day-old Cobb broilers were distributed in a completely randomized design with six treatments, five replicates, and 50 birds per experimental unit. Treatments used in both experiments were a basal diet with 0.10% Na+ (Experiment 1) or Cl- (Experiment 2) supplemented to result in diets with Na+ or Cl- levels of 0.10, 0.15, 0.20, 0.25 ,0.30, or 0.35%, respectively. In Experiment 1, results indicated an optimum Na+ requirement of 0.26%. Sodium levels caused a linear increase in arterial blood gas parameters, indicating an alkalogenic effect of Na+. The hypertrophic area of growth plate in the proximal tibiotarsi decreased with Na+ levels. The TD incidence decreased with increases in dietary Na+. Litter moisture increased linearly with sodium levels. In Experiment 2, the Cl- requirement was estimated as 0.25%. Chloride levels caused a quadratic effect (P ≤ 0.01) on blood gas parameters, with an estimated equilibrium [blood base excess (BE) = 0] at 0.30% of dietary CT-. No Cl- treatment effects (P ≥ 0.05) were observed on litter moisture or TD incidence. The best DEB for maximum performance was 298 to 315 mEq/kg in Experiment 1 and 246 to 264 mEq/kg in Experiment 2. We concluded that the Na+ and Cl- requirements for optimum performance of young broiler chickens were 0.28 and 0.25%, respectively.

Tele-education environment and didactic iconography in sexual transmitted diseases

The field of advertising has developed techniques of mass communication and rapid transmission of information. Among these techniques are slogans and iconographies. This work used such tools for educational purposes and was divided into three modules. Module 1 evaluated the use of simplified textual communication (slogans) about the subject Sexually Transmitted Diseases. It was applied to third-year medical students, which had not taken the course on this theme. To evaluate the impact of the textual communication form (slogans), long phrases were elaborated in scientific language containing 15 topics. From these long phrases, compact phrases were developed using techniques of the advertising area for elaboration of slogans. Three forms of didactic material about that theme were developed. The first form consisted of long phrases in descriptive topics, using scientific language. The second was constituted exclusively of compact phrases in the form of slogans, and the third was composed of the combination of the two previous forms. Then, 10 multiple-choice questions were elaborated and applied in two phases. In the first phase, application occurred immediately after the reading of didactic texts; in the second phase, it happened 60 days after the reading. For statistical analysis, Snedcor's F test was used for analysis of variance, at 5% significance level. There was an increase in memorization by students who read the material containing the association between long phrases and slogans, which indicates that the latter, when used as an auxiliary model of learning, can bring significant benefits for education. Module 2 consisted of analyzing the elaboration of educational videos produced in graphic computing (called iconographies) for development of dynamic communication means. The theme Hair Cycle was utilized. Viability of high quantities of information in few minutes of animation could be demonstrated together with the advantage of presenting the process in a dynamic form without wasting scientific details. Module 3 presented the inclusion of slogans in educational videos produced in graphic computing about the Hair Cycle, showing a new tool for rapid and efficient transference of data. Slogans and iconographies, when utilized in educational material, can bring significant benefits for the student's learning.

Chorismate synthase: An attractive target for drug development against orphan diseases

The increase in incidence of infectious diseases worldwide, particularly in developing countries, is worrying. Each year, 14 million people are killed by infectious diseases, mainly HIV/AIDS, respiratory infections, malaria and tuberculosis. Despite the great burden in the poor countries, drug discovery to treat tropical diseases has come to a standstill. There is no interest by the pharmaceutical industry in drug development against the major diseases of the poor countries, since the financial return cannot be guaranteed. This has created an urgent need for new therapeutics to neglected diseases. A possible approach has been the exploitation of the inhibition of unique targets, vital to the pathogen such as the shikimate pathway enzymes, which are present in bacteria, fungi and apicomplexan parasites but are absent in mammals. The chorismate synthase (CS) catalyses the seventh step in this pathway, the conversion of 5-enolpyruvylshikimate-3-phosphate to chorismate. The strict requirement for a reduced flavin mononucleotide and the anti 1,4 elimination are both unusual aspects which make CS reaction unique among flavin-dependent enzymes, representing an important target for the chemotherapeutic agents development. In this review we present the main biochemical features of CS from bacterial and fungal sources and their difference from the apicomplexan CS. The CS mechanisms proposed are discussed and compared with structural data. The CS structures of some organisms are compared and their distinct features analyzed. Some known CS inhibitors are presented and the main characteristics are discussed. The structural and kinetics data reviewed here can be useful for the design of inhibitors. © 2007 Bentham Science Publishers Ltd.