Effect of aging on the availability of amino acids from an immune-enhancing diet (IED) after a surgical stress in rats.

BACKGROUND & AIMS: Dietary amino acid (AA) requirements increase after a surgical stress while the systemic AA availability from the diet decreases with age, due to splanchnic sequestration. While immune-enhancing diets (IEDs) have been recommended for the nutritional management of surgical patients, the systemic bioavailability of their AA supply has not been evaluated in elderly surgical patients. This was determined in surgically-stressed IED-fed aged rats. METHODS: Thirty-four 5-month- or 21-month-old male Sprague-Dawley rats were used. After a gastrostomy and placement of a jugular vein catheter and a one-week recovery period, the animals underwent two 24 h-enteral feedings with an arginine-enriched IED (Impact®, Nestlé Health Science) before (healthy state) and 18 h after a standardized laparotomy, used as a model of surgical stress. During enteral nutrition, blood samples were repeatedly collected to measure plasma AA bioavailability (incremental areas under the curve) at 2, 5 and 24 h. Surgical stress was evaluated from urinary catecholamines and plasma protein profile. RESULTS: Whatever the age or stress situation, IED feeding was associated with decreased plasma glycine and increased alanine, proline and arginine. Aging was mainly associated with a delayed plasma AA accumulation in the first hours after the initiation of enteral nutrition. Stress was associated with higher plasma arginine increase and lower histidine, methionine, phenylalanine and tyrosine accumulation. Age and stress interactions seem limited. CONCLUSIONS: AA bioavailability from an arginine-enriched IED seems to be maintained whatever age and stress situation. Aging appears to be mainly associated with a delay in plasma AA accumulation probably related to age-associated splanchnic sequestration of AAs. Additional effects of surgical stress per se seem limited.

Effect of citrulline on muscle functions during moderate dietary restriction in healthy adult rats.

Low calorie diets are designed to reduce body weight and fat mass, but they also lead to a detrimental loss of lean body mass, which is an important problem for overweight people trying to lose weight. In this context, a specific dietary intervention that preserves muscle mass in people following a slimming regime would be of great benefit. Leucine (LEU) and Citrulline (CIT) are known to stimulate muscle protein synthesis (MPS) in post-prandial and post-absorptive state, respectively. This makes them interesting bioactive components to test in the context of dietary restriction. We tested the concept of combining LEU and CIT in adult female rats. We postulated that the sequential administration of LEU (mixed in chow) and CIT (given in drinking water before a rest period) could be beneficial for preservation of muscle function during food restriction. Sixty female rats (22 weeks old) were randomized into six groups: one group fed ad libitum with a standard diet (C) and five food-restricted groups (60 % of spontaneous intake for 2 weeks) receiving a standard diet (R group), a CIT-supplemented diet (0.2 or 1 g/kg/day, CIT0.2 group and CIT1 group, respectively), a LEU-supplemented diet (1.0 g/kg/day) or a CIT + LEU-supplemented diet (CIT + LEU 1.0 g/kg/day each). At the end of the experiment, body composition, muscle contractile properties and muscle protein synthesis (MPS) rate were studied in the tibialis anterior muscle. Dietary restriction tended to decrease MPS (R: 2.5 ± 0.2 vs. C: 3.4 ± 0.4 %/day, p = 0.06) and decrease muscle strength (R: 3,045 ± 663 vs. C: 5,650 ± 661 A.U., p = 0.03). Only CIT administration (1 g/kg) was able to restore MPS (CIT1: 3.4 ± 0.3 vs. R: 2.5 ± 0.2 %/day, p = 0.05) and increase muscle maximum tetanic force (CIT1: 441 ± 15 vs. R: 392 ± 22 g, p = 0.05) and muscle strength (CIT1: 4,259 ± 478 vs. R: 3,045 ± 663 A.U., p = 0.05). LEU had no effect and CIT + LEU supplementation had few effects, limited to adipose mass and fatigue force. The results of this study highlight the ability of CIT alone to preserve muscle function during dietary restriction. Surprisingly, LEU antagonized some effects of CIT. The mechanisms involved in this antagonistic effect warrant further study.

Evaluation of biohazards in dehydrated biofilms on foodstuff packaging.

Plastic materials used for food packaging are clean but not sterile when the food is just packaged. Accidental wet contamination may occur at every moment between packaging and opening by the consumer: on polyethylene (PET), bacteria may adhere strongly and constitute a biofilm in less than 24 h. By rolling on themselves, PET sheets may contaminate food. We tried to show that contact with salted foodstuffs favoured microbial recovery. Four strains were chosen to perform biofilms on PET: Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa and Escherichia coli. Biofilms were dried up 24 h. Biofilm bacteria were stressed by adhesion, by starvation and by dehydration. However, they were capable of recovery in salted solutions or media, probably because one (or more) stress protected them against another stress. Stress was demonstrated by stress protein production, by mean of electrophoresis, and membrane lesions by mean of flow cytometry. Stress recovery was performed in aqueous salted solutions or salted brain-heart infusion with NaCl 9, 15, 20 and 30 g/l. Staphylococci were more sensitive to these stresses and recovery was a function of salt concentration. Gram-negative bacteria were little affected by stresses; salt effects were less important. If all these biofilms were capable of recovery from stresses in salted media, flexible PET could possibly lead to a health hazard when it is used for wet salt meats, e.g.

[Infection on foreign material: bacterial colonization of ureteral endoprostheses]. / Infection sur matériel étranger: colonisation bactérienne des endoprothèses urétérales.

Infection on foreign body: bacterial colonization of ureteric stents. The most frequent cause of the early removal of ureteric endoprostheses (double J) is generally due to bacterial colonization. In order to prevent or to restrict the prosthesis colonization, it is necessary to understand the major steps and the factors influencing the colonization. This is the reason why we aimed to extract the most relevant parameters influencing the bacterial colonization from the observations made in vivo thanks to in vitro analyses. We have studied in vivo the relationship between the bacterial colonization of the endoprostheses, the urinary infections and the antibiotherapy. In vitro, we have defined the conditions promoting the primary adhesion of the most frequently isolated bacteria on endoprostheses. Surface properties of bacteria and materials have been compared to:--the bacterial count of infected double J samples with respect to bacterial species,--the bacterial count of the infected samples with respect to pH and Ca2+, Mg2+ concentration. The results show a great variability of the biomaterial surface properties which could be optimized, the fact that the urinary medium acidification could lower the bacterial adhesion and the ambiguous role of Ca2+ and Mg2+ ions which is discussed in this paper. In the case of in vivo analyses, the conflicting results between leukocyturia and bacteriuria lead to the detection of the bacterial colonization under antibiotic treatment. The characterized urinary infection must warn the risk of pyelonephritis.

The role of p53 as a transcription factor in the induction of apoptosis.

The tumor suppressor gene p53 plays a major role in the protection of cells from DNA damage. Activation of the protein in response to irradiation or genotoxic agents, and possibly by other signals, results in growth arrest at the G1 phase of the cell cycle or in apoptosis. While it has been shown that the ability of p53 to function as a sequence-specific transcriptional activator is necessary for the induction of growth arrest, the mechanism of p53-mediated apoptosis is not clear yet. In the present report we summarize the results obtained by several groups concerning p53-mediated apoptotic pathway. We suggest that p53 may induce apoptosis via a complex network of interacting pathways, which may be transcriptionally dependent or independent, depending on external signals and on the cellular context. Whatever the mechanisms are, the outcome-cell death by apoptosis-is a key function of the tumor suppressor p53.

Microorganisms as a tool of studying copper metal ions--induced changes in electrophysical cell properties.

Microorganisms are very sensitive to the effect of metal ions and a deficit or an excess of essential metal ions may induce some cell disturbances. The main objective of this research is to evaluate the effect of toxic metal ions. Doing that, the effects of metal ions on the electrophysical properties of the cell membrane, probed by electroorientational spectroscopic techniques and analysis of the electrophoretic mobility of cells have been chosen for investigation. The purpose of this work is to determine the optimal conditions of investigation of different kinds of microorganisms exposed to Cu. pH had a great influence on the electrophysical changes induced by Cu and this may be related to the speciation of the metal, according to its degree of hydroxylation. It will now be possible to compare the effects of toxic metal ions (Cd, Hg, Pb, etc..) or therapeutic metal ions (Pt, Ga, Ru, Ti, Sn, etc..) with those observed for Cu, in the conditions optimized by this study.

[In vivo and in vitro analysis of the ability of urinary catheter to microbial colonization]. / Analyse in vivo et in vitro de l'aptitude á la colonisation microbienne des sondes urinaires.

Bacterial adhesion to biomaterials is a complex phenomenon involving numerous factors. The ability to reduce urinary catheters infections simply by general hygiene and asepsis is low: an ascending colonization cannot be avoided. This will lead to a clinical infection only if several factors favour the bacterial adhesion or the bacterial coaggregation and the feeding of the bacterial biofilm. Among the many factors involved in bacterial adhesion, we focused in this paper on the physical parameters of surface hydrophobicity of the urinary catheters (Van der Waals and acido-basic forces) and the surface hydrophobicity of the bacteria (BATH and zeta potential). We also compared scanning electron microscopy (SEM) of in vivo and in vitro infected urinary catheters. We provided evidence that the more hydrophobic the bacteria, the more they are able to colonize hydrophobic materials, whereas hydrophilic cells are able to colonize hydrophilic materials more easily. Some biomaterials were found to display an irregular texture of hydrophobic and hydrophilic areas: they favour both types of adhesion. Moreover the divalent cations (MgII) drastically increased the bacterial coaggregation and favour bacterial growth within the biofilm. Finally, an increase in urinary pH and ionic strength increases the colonization risk. Consequently, choice of urinary catheter biomaterials is essential as patient hygiene and diet in order to avoid clinical infections.

FT-IR spectroscopy study of perturbations induced by antibiotic on bacteria (Escherichia coli).

Fourier transform infrared spectroscopy (FT-IR) is an analysis method which over the spectral absorption, gives information about the molecular structures of systems. Recently, this method is widely used to the investigation of complex systems like cells and bacteria. Characteristic of FT-IR spectrum of bacteria depend closely to physiological and culture parameters. In this study, the infrared bands of intact bacteria are first tentatively attributed to the contribution of the cellular components. Secondly are compared the FT-IR spectra of Escherichia coli bacteria before and after treatment at sub-inhibitrice concentrations (< or = MIC) at penicillin A, penicillin G and nalidixic acid. The observed spectral perturbations are closely depending on the antibiotic treatment and are observed even if bacterial cell mass is far away from cell death. On the other hand, this spectral changes are related to the known mode of action of the used antibiotic.

[Prevention of cariogenic dental plaque. Study of the structures implicated in the adhesion and coaggregation in Streptococcus mutans and Streptococcus sobrinus]. / Prévention de la plaque dentaire cariogène étude des structures impliquées dans l'adhésion et la coagrégation chez Streptococcus mutans et Streptococcus sobrinus.

Cariogenic dental plaque may be assimilated to a biofilm resulting from the adhesion of S. mutans, then from the coaggregation of other streptococci, or other genus. We used a static monospecific biofilm model. Supports or bacteria were treated with inhibitors before adhesion in order to clarify the nature of adhesins responsible for the primary adhesion of S. mutans and S. sobrinus on Tygon. To determine the bindings of coaggregation, inhibitors were applied on one-day-old biofilms. Analysis of effects were performed by automatic inoculator Spiral (Interscience) for microbiological methods, and by SEM JEOL 5400 LV for microscopic methods. In the aim of preventing adhesion and coaggregation, different traps were assayed:sugars, chemical inhibitors such as F- and EDTA salts. Of these, only the latter showed efficiency. This confirmed the role of bivalent mineral ions and electrostatic attraction forces in the adhesion and coaggregation of streptococci.

Monitoring of bacterial growth and structural analysis as probed by FT-IR spectroscopy.

Fourier-transform infrared spectroscopy was used to explore structural changes in bacteria under different incubation conditions. In particular, differences between Bradyrhizobium japonicum (BRJ) grown in liquid and on solid media were investigated, as well as the rearrangement of BRJ after transfer from one medium to the other. The FT-IR absorption bands located between 1200 and 900 cm-1 region, vary in spectral shape and intensity when BRJ were suspended in solution medium or plated on solid medium. In agreement with the electronic micrograph data, these spectroscopic changes are due to the changes involving the bacterial wall (peptidoglycan) when BRJ are plated in agar medium. By means of this FT-IR ultrastructural study of Bradyrhizobium japonicum bacteria, it has been possible to follow and to evaluate the rate of the molecular change in bacteria without any destructive interference. This indicates that FT-IR spectroscopy can prove to be a valuable technique in the monitoring of metabolic events in bacterial cells relevant to agriculture as well as environmental and health sciences.

[Synergism of the combination of enzymes or surfactants and a phenolic disinfectant on a bacterial biofilm]. / Synergie de l'association d'enzymes ou de surfactants et d'un désinfectant phénolique sur un biofilm bactérien.

Disrupting bacterial biofilms is necessary for a wide application domains such as reusable medical devices, or systems of pipes for water or fluids in cosmetics, food and chemicals industry. Bacterial cells embedded in a biofilm are less susceptible to disinfectants than suspended cells. This property is referable to the structure of the biofilm itself. The gangue of exopolymers and the thickness of a 5-day-old biofilm of Escherichia coli (more than 200 layers of bacteria), contribute to this decrease of susceptibility. The present work deals with the release of an Escherichia coli biofilm by the sequential action of enzymes and a phenolic disinfectant on the one hand, and by the sequential or simultaneous action of surfactants and the previous disinfectant on the other hand. The decrease of bacteria count per mm2 and the Scanning Electron Microscope observations exhibited a synergic action in every case. Nevertheless, Escherichia coli biofilms quickly reconstructed even after exposition to the previous treatment.

Antiseptic activity of some antidental plaque chemicals on Streptococcus mutans biofilms.

S. mutans is one of the bacterial species involved in the initiation of cariogenic dental plaque. In situ, antiseptic activity evaluation of dental plaque chemicals is fairly difficult. A dynamic biofilm model of S. mutans on Tygon support is suitable for the quantification of antiseptic activity and the comparison of commercial chemical formulations, submitted or not to the french standards and Pharmacopea. Some of these chemicals, especially quaternary ammonia solutions, do not reduce living bacteria counts by 10E5 as AFNOR specifies. In the case of dental plaque, antiseptic activity is probably not the only relevant parameter: biofilm dissociation is rather more relevant because the dead or living bacteria adhere to the teeth, favouring the establishment of a new active biofilm. Two proceedings can be followed: support surface can be modified, or bacteria exopolymers synthesis can be inhibited to prevent cell adherence; the disruption of the biofilm glycocalyx can be achieved with enzymes or surfactants.

[Comparison of the activity against Streptococcus mutans of a chlorhexidine-based antiseptic as a suspension or as a biofilm]. / Comparaison de l'activité d'un antiseptique à base de chlorhexidine sur Streptococcus mutans en suspension et en biofilm.

When bacteria colonize a surface they form a biofilm whose susceptibility to anti-microbials is different from that of the same bacterial species forming a homogeneous suspension in a liquid. This study investigated colonization of an inert solid phase (Tygon) with Streptococcus mutans ATCC 25,175--one of the strains involved in the initiation of cariogenic dental plaque--in a continuous flow of fresh medium sufficiently diluted so as to preclude growth of suspended bacteria. Only those bacteria which adhered to the solid phase grew, forming a biofilm. The antiseptic activity of Eludril (0.1% chlorhexidine) on this biofilm under dynamic (flowing medium) and static (stagnant medium) conditions was studied by comparison with the same strain in a suspension (in compliance with the AFNOR NF T 72-150 norm) and in a confluent culture on a filtering membrane. The biofilm was less susceptible under dynamic than under static conditions; under both conditions, the biofilm was less susceptible than the suspension. According to this model, the concentration of antiseptic recommended by the manufacturer according to studies using AFNOR norm NF T 72-150 (bacteria in a suspension) may be inadequate for bacteria adhering to tooth surfaces or gingival mucosa.

Tratamiento antibiótico preventivo en cirugía urológica / Antibiotical treatment in urologic surgery

En estudio retrospectivo de 151 pacientes sometidos a RTU y con orinas estériles, observan un 38% sde infecciones post-operatorias. En trabajo prospectivo se ensaya una dosis única de Mezlocillina, antibiótico efectivo sobre Gr (+), Gr (-) y Pseudomonas. Se observa una sóla infección (2%) en el grupo tratado durante el primer día, contra 9 casos (18%) en el grupo no tratado. Al quinto día de control, la infección está presente en 20 casos (41,6%) y 25 casos (49%) respectivamente

[Bone and blood levels of azlocillin after intra-arterial injection in the dog]. / Taux osseux et sanguins de l'azlocilline après injection intra-artérielle chez le chien.

Pharmacokinetic values of azlocillin--a semisynthetic penicillin of the ureidopenicillin family--were determined in dogs after injection of the drug into the right femoral artery by measuring plasma concentrations in the right femoral artery and left femoral vein and tissue concentrations in ipsi- and contralateral bones. Two different doses of azlocillin were administered, one group of dogs receiving 100 mg X kg-1 by fast (1 min) bolus injection and the other group receiving 250 mg X kg-1. The drug was assayed by high performance liquid chromatography in reverse phase, which proved to be the simplest, most rapid and most sensitive method. Changes in plasma and bone tissue concentrations after intra-arterial injection were interpreted as order one kinetics with a two-compartment open model. Following the 100 mg/kg dose, concentrations versus time corresponded to the following equations: --Arterial blood (mg/l) = 7570 e-0.15 t (min) + 240 e-0.017 t --Venous blood (mg/l) = 553 e-0.13 t + 222 e-0.016 t --Bone tissue (micrograms/g) = 15 e-0.0115 t (elimination phase). Distribution and elimination constants were identical in arterial and venous blood. Distribution half-life was 5 +/- 1 min in arterial blood and biological half-life was 42 +/- 11 min. However, bioavailability was comparatively lower in veins due to higher levels in arteries during the distribution phase. Bone tissue concentrations were much lower than plasma concentrations and were identical on both sides despite high arterial levels on the ipsilateral side during the distribution phase. Bone concentrations diminished rapidly and elimination kinetic values were the same in bone and in plasma. It is concluded that bone tissue concentrations of azlocillin after intra-arterial injection are highest during the distribution phase.