[Isolated girdle weakness: expansion of the phenotypic spectrum of the MERRF 8344A>G mutation of mitochondrial DNA]. / Debilidad aislada de cinturas: ampliacion del espectro fenotipico de la mutacion MERRF 8344A>G del ADN mitocondrial.

INTRODUCTION: The differential diagnosis of diseases that are accompanied by adult-onset girdle weakness is broad and includes motor neurone, neuromuscular junction or muscular diseases. The 8344A>G mutation of the MTTK gene of mitochondrial DNA usually presents with involvement of multiple organs associated (or not) with girdle weakness. To date no cases of isolated girdle weakness have been reported as the presenting symptom of this mutation. CASE REPORT: A 57-year-old male, with a four-year history of isolated clinical signs of progressive girdle weakness. He is the brother of a 59-year-old woman with the same clinical features. Muscular biopsy played a decisive role in the diagnosis and was characteristic of mitochondrial myopathy. The genetic analysis revealed the 8344A>G mutation of the MTTK gene of mitochondrial DNA. CONCLUSIONS: The 8344A>G mutation of mitochondrial DNA can be associated with clinical signs and symptoms of adult-onset girdle weakness, and must therefore be included as part of its differential diagnosis.

TITLE: Debilidad aislada de cinturas: ampliacion del espectro fenotipico de la mutacion MERRF 8344A>G del ADN mitocondrial.Introduccion. El diagnostico diferencial de los trastornos que cursan con debilidad de cinturas de inicio en la edad adulta es amplio e incluye enfermedades de neurona motora, union neuromuscular o musculo. La mutacion m.8344A>G del gen MTTK del ADN mitocondrial suele presentarse con afectacion de multiples organos asociada o no a una debilidad de cinturas. No se han descrito hasta el momento casos de debilidad de cinturas aislada como sintoma de presentacion de esta mutacion. Caso clinico. Varon de 57 años, con clinica aislada de debilidad progresiva de cinturas, de cuatro años de evolucion. Hermano de una mujer de 59 años con la misma sintomatologia. La biopsia muscular fue decisiva en el diagnostico y es caracteristica de una miopatia mitocondrial. El analisis genetico objetivo la mutacion m.8344A>G del gen MTTK del ADN mitocondrial. Conclusiones. La mutacion 8344A>G del ADN mitocondrial puede cursar con un cuadro aislado de debilidad de cinturas de inicio en el adulto, por lo que debe de formar parte del diagnostico diferencial de este.

Influence of Mitochondrial Genetics on the Mitochondrial Toxicity of Linezolid in Blood Cells and Skin Nerve Fibers.

The antibiotic linezolid is a ribosomal inhibitor with excellent efficacy. Although the administration period has been reduced to 28 days, side effects, usually of hematologic or neuropathic origin, are still reported due to secondary inhibition of mitochondrial protein synthesis. Susceptibility to linezolid toxicity remains unknown. Therefore, the objective of this study was to gain an understanding of clinical heterogeneity in response to identical linezolid exposures through exhaustive examination of the molecular basis of tissue-dependent mitotoxicity, consequent cell dysfunction, and the association of mitochondrial genetics with adverse effects of linezolid administered for the recommended period. Peripheral blood mononuclear cells (PBMC) and skin nerve fibers from 19 and 6 patients, respectively, were evaluated before and after a 28-day linezolid treatment in order to assess toxic effects on mitochondria and cells. Mitochondrial DNA haplotypes and single nucleotide polymorphisms (SNPs) in ribosomal sequences where linezolid binds to mitochondrial ribosomes were also analyzed to investigate their genetic contributions. We found that linezolid reduced mitochondrial protein levels, complex IV activity, and mitochondrial mass in PBMC and was associated with a trend toward an increase in the rate of apoptosis. In skin tissue, mitochondrial mass increased within nerve fibers, accompanied by subclinical axonal swelling. Mitochondrial haplogroup U, mutations in 12S rRNA, and the m.2706A→G, m.3197T→C, and m.3010G→A polymorphisms in 16S rRNA showed a trend toward an association with increased mitochondrial and clinical adverse effects. We conclude that even when linezolid is administered for a shorter time than formerly, adverse effects are reported by 63% of patients. Linezolid exerts tissue-dependent mitotoxicity that is responsible for downstream cellular consequences (blood cell death and nerve fiber swelling), leading to adverse hematologic and peripheral nervous side effects. Multicentric studies should confirm genetic susceptibility in larger cohorts.

Identification and biochemical characterization of the novel mutation m.8839G>C in the mitochondrial ATP6 gene associated with NARP syndrome.

Mutations in the ATP6 gene are reported to be associated with Leber hereditary optic neuropathy, bilateral striatal necrosis, coronary atherosclerosis risk and neuropathy, ataxia and retinitis pigmentosa (NARP)/maternally inherited Leigh syndromes. Here, we present a patient with NARP syndrome, in whom a previously undescribed mutation was detected in the ATP6 gene: m.8839G>C. Several observations support the concept that m.8839G>C is pathogenically involved in the clinical phenotype of this patient: (1) the mutation was heteroplasmic in muscle; (2) mutation load was higher in the symptomatic patient than in the asymptomatic carriers; (3) cybrids carrying this mutation presented lower cell proliferation, increased mitochondrial DNA (mtDNA) copy number, increased steady-state OxPhos protein levels and decreased mitochondrial membrane potential with respect to isogenic wild-type cybrids; (4) this change was not observed in 2959 human mtDNAs from different mitochondrial haplogroups; (5) the affected amino acid was conserved in all the ATP6 sequences analyzed; and (6) using in silico prediction, the mutation was classified as 'probably damaging'. However, measurement of ATP synthesis showed no differences between wild-type and mutated cybrids. Thus, we suggest that m.8839G>C may lower the efficiency between proton translocation within F0 and F1 rotation, required for ATP synthesis. Further experiments are needed to fully characterize the molecular mechanisms involved in m.8839G>C pathogenicity.

Kidney androgen-regulated protein transgenic mice show hypertension and renal alterations mediated by oxidative stress.

BACKGROUND: Kidney androgen-regulated protein (KAP), a proximal tubule androgen-regulated gene, codes for a protein of unknown function. METHODS AND RESULTS: To investigate the consequences of KAP overexpression in kidney, we produced KAP transgenic mice and performed microarray expression analyses in kidneys of control and transgenic males. Downregulation of the androgen-sensitive Cyp4A14 monooxygenase gene in KAP transgenic mice prompted us to analyze blood pressure levels, and we observed that transgenic mice were hypertensive. Inhibition of 20-hydroxyeicosatetraenoic acid synthesis by N-hydroxy-N'-(4-n-butyl-2-methylphenyl) formamidine (HET0016) reduced the increased 20-hydroxyeicosatetraenoic acid levels in urine and normalized arterial pressure in transgenic mice, as did the NADPH oxidase inhibitor apocynin. Increased oxidative stress in transgenic mice was demonstrated by (1) enhanced excretion of urinary markers of oxidative stress, 8-iso-prostaglandin F2alpha, 8-hydroxydeoxyguanosine, and thiobarbituric acid-reacting substances; (2) augmented mitochondrial DNA damage and malondialdehyde levels in kidneys; and (3) diminished catalase and glutathione peroxidase activity in transgenic kidneys. Mice exhibited renal defects that included focal segmental glomerulosclerosis, proteinuria, glycosuria, and fibrosis. CONCLUSIONS: Taken together, these results indicate that KAP expression is critical for cardiovascular-renal homeostasis maintenance and that hypertension is associated with increased oxidative stress. This is the first report showing that overexpression of an androgen-regulated, proximal tubule-specific gene induces hypertension. These observations may shed light on the molecular pathophysiology of gender differences in the prevalence and severity of hypertension and chronic renal disease.

Mitochondrial DNA oxidation and manganese superoxide dismutase activity in peripheral blood mononuclear cells from type 2 diabetic patients.

AIM: To investigate the balance between parameters of oxidative stress and antioxidant defences in the mitochondria of peripheral blood mononuclear cells (PBMCs) of type 2 diabetic patients with late complications. METHODS: Ten type 2 diabetic patients with late diabetic complications and 10 age-matched healthy volunteers (controls) were prospectively recruited. Mitochondrial DNA (mtDNA) oxidative damage and mtDNA content were measured as indices of oxidative stress. Manganese superoxide dismutase (MnSOD) activity has been used as an index of mitochondrial antioxidant defence. Mitochondrial respiratory-chain function (cytochrome C oxidase activity) was also assessed. RESULTS: Mitochondrial DNA (mtDNA) oxidation was significantly higher in the PBMCs of diabetic patients than in control subjects (P<0.0001) and, although mtDNA content was lower in the diabetic group, this was not statistically significant. MnSOD activity was significantly increased in PBMCs of type 2 diabetic patients compared with healthy controls (1366+/-187 versus 686+/-167 U/g of protein; P=0.01), and was related to mtDNA oxidative damage. No differences in mitochondrial respiratory-chain function were found between diabetic patients and controls. CONCLUSION: PMBCs from type 2 diabetic patients with late diabetic complications exhibit high mtDNA oxidative damage. The degree of mtDNA oxidation was associated with an increase in MnSOD as an adaptive response to oxidative stress. The consequences of mtDNA oxidative damage on PBMC function and the progression of diabetic complications remain to be elucidated.

Nutritional modulation of protein metabolism after gastrointestinal surgery.

BACKGROUND: The metabolic response to surgery includes alterations in protein metabolism, resulting in a net loss of proteins. Protein hypercatabolism is considered an unavoidable consequence of injury, and an important source of morbidity and mortality. Our purpose was to determine the effect of nutrition on protein metabolism following gastrointestinal surgery, and to elucidate whether postoperative protein loss can be prevented with adequate nutritional support. METHODS: Patients who had undergone gastrointestinal surgery were given four different parenteral nutritions with increasing glucose, lipid and amino acid content during the 7 days following surgery. Nitrogen balance, protein synthesis and protein breakdown were determined using in vivo stable isotope labelling. Other metabolites (3-methylhistidine, creatinine, urea, cortisol, glucose, insulin, amino acids and C-reactive protein) were measured. RESULTS: A nutrition-dependent alteration of protein metabolism was found in response to surgical injury. Nutrition modified nitrogen balance, whole-body protein breakdown and, to a lesser extent, whole-body protein synthesis and muscle protein breakdown. The low-energy parenteral nutrition without amino acids produced a negative nitrogen balance (postoperative day 7=-0.381 g protein kg(-1)day(-1)) and important alterations in postoperative protein metabolism that did not normalize during the study period (day 7 protein synthesis=239% and protein breakdown 217% vs preoperative). Patients receiving the two low energy parenteral nutritions containing amino acids had a less negative nitrogen balance (day 7=-0.011 and -0.133 g protein kg(-1)day(-1)) and a transient increase in protein metabolism. The complete parenteral nutrition maintained, during all studied days, protein metabolism parameters within the preoperative reference range (synthesis day 2=92%, day 4=110% day 7=79%; breakdown day 2=85%, day 4=80%, day 7=76% vs preoperative) and a positive nitrogen balance (day 2=+0.0387, day 4=+0.578 and day 7=+0.227 g protein kg(-1)day(-1)). CONCLUSION: Complete nutritional support can prevent protein loss after gastrointestinal surgery and maintain protein metabolism without alterations.

Comparison of different techniques for purification of triamcinolone acetonide suspension for intravitreal use.

BACKGROUND: Intravitreal triamcinolone has increasingly been used for the treatment of oedematous and neovascular diseases and purification of triamcinolone suspension may be important in order to avoid the potential toxic effects of the vehicle. The aim was to evaluate different techniques used to reduce the solvent agent benzyl alcohol (9.9 mg/ml) from a commercially prepared triamcinolone acetonide suspension. METHODS: Different techniques were used to reduce the solvent agent benzyl alcohol: filter techniques using 0.22 mum or 5 mum pore size, and non-filter techniques using sedimentation or centrifugation. Quantification of triamcinolone acetonide and benzyl alcohol was performed by high pressure liquid chromatography (HPLC). RESULTS: Benzyl alcohol concentration was decreased significantly in all the techniques used compared with the original commercial suspension (p<0.05), with no significant differences among them. The reduction was approximately one tenth of its original concentration. However, triamcinolone acetonide concentration differed significantly depending on the method used. Centrifugation method showed no differences versus the original commercial solution; sedimentation technique reduced the expected dose only 25%; the filter technique using a 5 mum pore size membrane reduced the expected dose to one fourth, while the filter technique using a 0.22 mum pore size membrane reduced the expected dose to 45%. CONCLUSIONS: All the different techniques employed effectively reduced the concentration of benzyl alcohol. However, the final concentration of triamcinolone was much lower than expected using the filter techniques. The pore size membrane inversely influenced the final concentration, with part of the triamcinolone crystals probably being entrapped in the filter. Centrifugation is recommended as the best way of administering the drug.

Usefulness of short-lived proteins as nutritional indicators surgical patients.

BACKGROUND AND AIMS: Biochemical indicators are used to assess the adequacy of nutritional support given to postoperative patients. However, the metabolic alterations present in these patients diminish the efficiency of these indicators. The objective of this work is to determine the usefulness of short-lived proteins as indicators to assess the nutritional support administered to patients during the metabolic stress phase produced by surgery. METHODS: The nitrogen balance and plasma concentrations of transthyretin, retinol binding protein, and insulin-like growth factor-1 were determined in 24 patients who received 4 different nutritional regimens during 7 days after surgery. RESULTS: Transthyretin and retinol binding protein, although sensitive to nutritional intake (P<0.0005 and P<0.04 respectively), were strongly affected by the stress response (P<0.008 and P<0.0003 respectively), thus limiting their usefulness for nutrition assessment. Insulin-like growth factor-1 was not influenced by the stress response and was sensitive to the nutritional supply (P<0.0001). Insulin-like growth factor-1 was the only component that showed similar efficiency than nitrogen balance as nutritional indicator. CONCLUSIONS: Transthyretin and retinol binding protein are not adequate to assess the nutritional supply during the stress phase after surgery, while insulin-like growth factor-1 is a suitable indicator of the adequacy of recent intake in this situation, similar in performance to nitrogen balance.

Effect of starvation on organ blood flow in the senescent rat.

To investigate the amino acid requirements of the senescent rat, as part of a study directed toward nutritional support in the aged, it was necessary to determine amino acid levels in plasma and tissue, but also regional blood flow of the animals subjected to fast. Only this latter allows the determination of the amounts of each amino acid present in the tissue before starvation by extrapolation of values measured during starvation. As plasma and tissue amino acid had been previously determined, the aim of this study had been to measure regional blood flow in the liver, kidney, testis, spleen, stomach, small intestine and large intestine in senescent rats submitted to 1, 5, 9 and 15 days of starvation. Twenty-four-month-old male Wistar rats (n = 16) were divided into four groups (n = 4), and submitted to starvation for 1, 5, 9 and 15 days. Blood flow in the liver, kidney, testis, spleen, stomach, and small and large intestine was measured by injecting 0.5 ml of a microsphere solution (15 microns diameter) labelled with 57Co, 0.25 microCi/ml. Over the 15-day period studied, the response to starvation showed two distinct phases: an early effect (from day 1 to day 9) in which there were decreases in the weight of the organs and in organ blood flow, and a second phase (from day 9 to day 15) in which blood flow and organ weight were maintained. However, organ blood flow related to mass was not substantially affected by starvation. This implies that measurement of substrate plasma concentration alone can reliably reflect organ substrate flow.

Oxidative stress induces age-dependent changes in lymphocyte protein synthesis and second messenger levels.

Cumulative damage in cells from aged people could lead to a greater fragility against acute oxidative stress. The effects of acute oxidative stress on cell viability, cAMP and cGMP concentrations, and protein synthesis rates were studied in lymphocytes from 25 young and 26 elderly subjects. Lymphocytes were exposed to stress by hydrogen peroxide 25 micromol/l and incubated for 18 hours. Cell viability after stress was lower (p<0.0001, Student's t test) in cells from the elderly (63.4%) than in cells from the young donors (73.2%). The protein synthesis rate was also lower after stress (p<0.04, Mann-Whitney U test) in cells from the elderly (47.3% vs. non-stressed cells), than in cells from the young (82.19% vs. non-stressed cells). After oxidative stress, cAMP and cGMP concentrations showed no significant changes in cells from young subjects; there were, however, significant decreases in these cyclic nucleotides in cells from the elderly (p<0.008 for both nucleotides, paired Student's t test). There were no differences in basal cAMP or cGMP levels between the two groups. These results show that mortality and metabolic changes due to oxidative stress are greater in lymphocytes proceeding from elderly subjects than in those from young subjects.

Effect of oxidative stress on lymphocytes from elderly subjects.

1. Oxidative damage has been associated with ageing, but there is no agreement as to whether or not it is produced by a decrease in antioxidant defences with the ageing process. In purified lymphocytes from 47 healthy elderly (75.27 +/- 0.91 years) and 47 healthy young (29.87 +/- 0.53 years) volunteers, we studied the levels of antioxidant enzyme activity (superoxide dismutase, catalase and glutathione peroxidase), protein oxidative damage (as protein carbonyl content) and lysosomal proteolytic activity (cathepsins B, H and L), with and without exposure to oxidative stress produced by 25 mumol/l H2O2. 2. There were no differences in antioxidant enzyme activities in the stressed and non-stressed samples between the young and elderly subjects, indicating that there was no relationship between age and antioxidant enzyme activity even in oxidative stress. However, a dissimilar response to oxidative stress was observed in protein oxidative damage and cathepsin B and L activities, depending on the age of the donor. 3. With these results we conclude that oxidative stress produces greater protein oxidative damage and increased protein degradation in elderly subjects than in young ones; this effect cannot be attributed to dissimilar antioxidant enzyme responses to oxidative stress, since these did not differ between the two age groups.

Early curbing of protein hyper-catabolism in postoperative patients by nutritional support with glucose plus amino acids, but not with glucose alone.

This work attempts to determine if there are differences in protein metabolism in post-surgical patients who receive parenteral nutrition with amino acids plus glucose (G+AA) or conventional gluco-salinal solution (GS). Eighteen patients submitted to gastrointestinal surgery were randomized and double-blindly administered either G+AA (1 g AA/kg x d and 28 kJ/kg x d), or GS (28 kJ/kg x d). Protein metabolism was determined 12 h after surgery (day 0) and after 5 days of nutritional support. On day 0, protein breakdown was similarly elevated, with respect to reference values, in both groups (GS: 4.62 +/- 0.25; G+AA: 5.25 +/- 0.50 g prot/kg x d) as a result of surgical stress. These values increased significantly at day 5 (P < 0.03) with the administration of GS to 6.93 +/- 1.00 g prot/kg x d, while they decreased (P < 0.002, 3.30 +/- 0.42 g prot/kg x d) with G+AA. Protein synthesis was increased (5.69 +/- 0.86 g prot/kg x d) with GS (P < 0.02), and was decreased (2.79 +/- 0.44 g prot/kg x d) with G+AA (P < 0.0002). Both synthesis and breakdown were inside normal reference values after 5 days for group G+AA. In both groups, nitrogen balance did not change significantly at day 5 compared to day 0. G+AA is effective in curbing the hypermetabolism produced by postoperative stress, achieving normal protein metabolism in 5 days, while GS increases the protein breakdown and synthesis. Nitrogen balance does not detect these modifications of the protein metabolism. Undernutrition on prognosis is not yet fully recognized.

Modification of organ protein synthesis after surgical stress by low energy diets with different supplements.

We have studied the effects of hypocaloric diets with different supplements on liver and jejunal mucosa protein synthesis. The supplements assayed were medium chain triglycerides (diet MCT, with 50% carbohydrates: 25% long chain triglycerides (LCT): 25% medium chain triglycerides (MCT), standard amino acids), branched-chain amino acids (diet BCA, identical to control diet L50, with 15.3% of nitrogen replaced by branched-chain amino acids) and glutamine (diet GLN, identical to diet L50, with 15.3% of nitrogen replaced by glutamine). The control diet (L50) had 50% carbohydrates: 50% LCT and standard amino acids. The diets were assayed on 86 rats with femoral fracture immobilized by Kirschner pin insertion. Nutrition was administered for 4 days. On the fifth day, liver and jejunal mucosa protein synthesis was determined. A branched-chain amino acid supply in a proportion higher than 21.2% of amino acid nitrogen significantly decreased liver and jejunal mucosa protein synthesis, while the same amount of glutamine did not modify it. MCT had no effect on jejunal mucosa protein synthesis, while it was decreased significantly in the liver.

Modification of organ protein synthesis after surgical stress by low energy diets with different lipid/glucose ratios.

The aim of this work was to study the effects of low energy parenteral diets with different lipid/glucose ratios on rat liver and jejunal mucosa protein synthesis. The studied diets were: L0 (100% glucose, control diet), L25 (25% lipids: 75% glucose), L50 (50% lipids: 50% glucose) and L75 (75% lipids: 25 % glucose). All diets were isoenergetic and isonitrogenated, with a standard amino acid content. The diets were assayed in 93 rats with open femoral fracture immobilized by Kirschner pin insertion. The diets were administered for 4 days. On the fifth day, liver and jejunal mucosa protein synthesis were determined. Highest liver protein synthesis rates were obtained with the diet compositions: lipid/carbohydrate ratio: 25% lipids and 75% carbohydrates (expressed as energy ratio). A higher proportion of lipids significantly decreases liver protein synthesis (p <0.05). Jejunal mucosa protein synthesis followed the same pattern, with the same statistical differences.

[Amino acid pattern variation in animals after the administration of hypocaloric peripheral parenteral nutrition]. / Variación del patrón de aminoácidos en animales tras la administración de nutrición parenteral periférica hipocalórica.

The pattern of intracellular aminoacids may be reproduced individually and is different for each illness. This study seeks to measure alterations in this pattern in rats after the administration of hypocaloric peripheral parenteral nutrition (HPPN), group B, for five days, by comparing them with rats subjected to conventional fluidtherapy, group A, for the same period. HPPN tolerance was good in all cases. Our results show a higher tendency to glyconeogenesis in group A, measured by a reduction in the glycogenic aminoacids especially alanine. An increase in branched aminoacids was observed in group A due to an increase in proteolisis. The animals in group B showed a lesser reduction in the intracellular aminoacids necessary for glyconeogenesis, especially glycine. HPPN, was found, to alter the aminoacids pattern in fasting rats. The results might be corroborated in humans studies.

Protein synthesis in specific tissues during sepsis.

BACKGROUND: The hypothesis that fractional protein synthesis rates (Ks) are tissue-specific and bidirectional during sepsis was tested in an animal model. MATERIAL AND METHODS: Ks in liver, triceps muscle, and diaphragm were measured in septic (n = 27) and control rats (n = 26). Sepsis was induced by a reproducible model established in our laboratory (intraperitoneal injection of sterile NaOH 0.75 N at 0.075 ml/100 g of body weight). Ks were measured using the flooding-dose method in tissue obtained from the diaphragm, liver, and from the triceps muscle. RESULTS: In hepatic and diaphragmatic tissue, Ks were significantly higher in the septic animals (Ks: 112.2 +/- 8 and 5.4 +/- 1.9, respectively) than in control animals (Ks: 78.5 +/- 13 and 2.9 +/- 1.7, respectively). In the triceps, Ks were significantly lower in septic animals (Ks: 2.9 +/- 1.4) than in control animals (Ks: 5 +/- 1.8). CONCLUSION: The results suggest that in septic animals the rate of protein synthesis is enhanced in tissues of priority, such as the liver, and varies in response to differences in muscle activity.

Addition of glutamine does not improve protein synthesis and jejunal mucosa morphology in non-hypercatabolic stress.

To investigate the effect of glutamine-enriched total parenteral nutrition (TPN) on the protein synthesis and morphology of jejunal mucosa in non-hypercatabolic stress, sixty-two male Sprague-Dawley rats were subjected to surgical stress by femoral fracture. The rats were divided into 3 groups and received TPN for 8 days. One group received a standard amino acid solution without glutamine, the second group a standard solution enriched with glycine and glutamic acid, and the third group a standard solution enriched with glycyl-glutamine. All regimens were isocaloric and isonitrogenous-nitrogen (2.2 g/kg.day), glucose (150 Kcal/kg.day), and lipids (150 Kcal/kg.day). There were no statistically significant differences in jejunal mucosal thickness, DNA content, protein content, fractional synthesis rate or absolute protein synthesis among the groups after eight days of parenteral nutrition. In conclusion, the addition of glutamine to TPN did not influence either protein metabolism or morphology of the jejunal mucosa in non-hypercatabolic surgical stress.

Effect of medium chain triglycerides (MCT) on jejunal mucosa mass and protein synthesis.

The effects of medium chain triglycerides (MCT) on jejunal mucosa mass and protein synthesis were compared with results from previous experiments with rats fed by parenteral nutrition or enteral nutrition. Other published studies have also been analysed. Three experimental models were studied. In the traumatic model, production of a femoral fracture was followed by Kirschner pin insertion into the medullary canal of both fragments at reduction. (Forty ras were fed enteral nutrition and 93 were given parenteral nutrition.) A second model entailed resection under ether anaesthesia using the technique described by Higgins. (Fifty five rats were fed enteral nutrition and 28 with parenteral nutrition.) A third model entailed a terminolateral portocaval shunt under anaesthesia with pentobarbital. (Sixty nine rats were treated this way and then given enteral nutrition.) Proportions of medium chain/long chain triglycerides (LCT) were as follows: 0/100, 20/80, 40/60, 50/50, and 92/8 for enteral nutrition and 0/100, 30/70, 50/50, and 70/30 for parenteral nutrition. Faecal losses of alpha amino nitrogen, protein, total fats, and free fatty acids were analysed together with the quantitative intake, weight gain of the rats, jejunal mucosal mass, and protein synthesis in relation to the MCT proportion ingested or given by enteral nutrition or parenteral nutrition. From analysis of our results and those of others, several conclusions could be drawn. Firstly, the route of administration of MCT is extremely important and enterocytes might be considered one of the main target sites. Secondly, a high proportion of MCT (more than 80%) offers no advantage for jejunal mucosa and produces undesirable side effects. Thirdly, the effect of MCT on jejunal mucosal protein synthesis depends on the metabolic state. Finally, an increase in jejunal mucosal mass directly correlated with MCT concentrations, but no correlation was found between mass and protein synthesis. A positive correlation, however, between MCT proportion and enzyme activity (alkaline phosphatase and sucrase) in the brush border membrane was seen as well as a positive correlation with the concentration of phospholipids in the microvilli.