Protein Synthesis In Patients Research Articles

Abstract 2055: Delivery of oligonucleotide therapeutic drugs: Universal detection reagents specific for nucleic acid modifications

Abstract Nucleic acid therapies, including oligonucleotide therapeutics (ONTs) and vaccines, have increased in development, respectively, since their inception as an emerging drug modality and after the first mRNA COVID-19 vaccine was granted EUA approval in 2020. RNA-based therapies, such as antisense oligonucleotides (ASO), RNAi, aptamers, and mRNA, are designed to modulate protein synthesis in patients and have applications in oncology, gene therapy, and infectious disease prevention. Strategies for ONT drug delivery have advanced over recent years with the goal of overcoming limitations when targeting extrahepatic tissues. Improving the “drug-likeness” of ONTs is facilitated by chemical modifications to the sugar-phosphate backbone and/or nitrogenous bases. Modifications are designed to increase ONT drug stability, uptake, and efficacy, but may increase the likelihood of toxicity. First generation chemical modifications include phosphorothioate (PS) modification of the phosphate portion of the backbone, whereas second generation modifications include adding substituent groups to the 2’ position of the ribose portion of the backbone, specifically 2’-O-methyl (OME), 2’-O-methoxy-ethyl (MOE), and 2’-fluoro modifications of RNA. Third generation modifications include peptide nucleic acids (PNA), locked nucleic acids (LNA), morpholino phosphoroamidate (PMO) modifications, and others. We have developed, optimized, and validated a library of monoclonal antibody reagents that detect these chemical modifications independent of nucleic acid composition, structure, strandedness, configuration, or platform. These “universal detection reagents” can be used in (1) immunohistochemistry (IHC) to determine biodistribution, (2) immunofluorescence (IF) to determine intracellular localization, (3) ELISA to determine anti-drug antibody binding, immunogenicity, and drug ranking, (4) immunoprecipitation to determine protein binding in serum and/or tissue lysates, (5) in vivo cell culture studies to determine potency, and (6) other immunoassays. While in situ hybridization (ISH) is an established method for the detection and localization of ONTs in cells and tissues, this method has limitations that include lack of utility (as unique probes must be designed individually for each ONT), high costs in time and materials to produce probes, the inability of ISH to bind and detect short ONT drug sequences, and difficulties encountered when repeated sequences are present within the ONT sequence. Here we report differential sensitivity and specificity of panels of antibodies specific for PS and MOE modifications. We demonstrate the performance of these antibodies by various in vitro assays that show the utility of these specialty reagents to facilitate the collection of analytical data to support ADMET studies useful to ensure ONT approval by regulatory agencies. Citation Format: Todd Giardiello, Amy Anderson, Renee Salvo, Carl Ascoli. Delivery of oligonucleotide therapeutic drugs: Universal detection reagents specific for nucleic acid modifications [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2055.

Features of protein synthesis in patients with chronic critical illness

Features of protein synthesis in patients with chronic critical illness

Determining ideal balance among branched-chain amino acids in medical formula for Propionic Acidemia: A proof of concept study in healthy children

BackgroundPropionic Acidemia (PROP) is an inherited metabolic disorder, with defect in the enzyme propionyl-CoA carboxylase (PCC) which catalyzes catabolism of two of the branched-chain amino acids (BCAA), valine, isoleucine. Nutritional management in PROP depends on dietary protein restriction and consumption of medical formula depleted of the offending amino acids. Recently, concerns have been raised about medical formula due to imbalanced content of BCAA (high leucine – another BCAA, and no valine/isoleucine), which negatively impacts plasma concentrations of BCAA, and growth in children with PROP. Objectives and methodsTo determine an optimal BCAA ratio at which total body protein synthesis is optimized in healthy children using the indicator amino acid oxidation method (oxidation of L-13C-Phenylalanine to 13CO2). This was accomplished by reducing leucine intake gradually from the current high dose in medical formula, in order to compare protein synthesis, under different BCAA ratios. ResultsA total of 8 healthy children were studied, completing 42 study days. Significant differences in F13CO2 with different BCAA ratios were found. BCAA ratio (leucine: isoleucine: valine) 1:0:0 was associated with the highest F13CO2 (low protein synthesis) compared to other ratios. By reducing leucine intake, and isoleucine and valine at minimum PROP recommendations, BCAA ratio between1:0.26:0.28 to 1:0.35:0.4 was associated with optimal protein synthesis. ConclusionBCAA ratio of 1:0:0, present in medical formula limited total body protein synthesis. A balanced BCAA ratio was found between 1:0.26:0.28 and 1:0.35:0.4 (leucine:isoleucine:valine). Future research is needed to test this optimal BCAA ratio for optimizing protein synthesis in patients with PROP. SynopsisThe article describes a proof-of-concept study done on healthy school-aged children testing different ratios of branched chain amino acid (BCAA, leucine:isoleucine:valine), in order to determine an optimal ratio at which total body protein synthesis is improved and has implications for dietary management of children with Propionic Acidemia (PROP).

Protein synthesis levels are increased in a subset of individuals with fragile X syndrome.

Fragile X syndrome (FXS) is a monogenic form of intellectual disability and autism spectrum disorder caused by the absence of the fragile X mental retardation protein (FMRP). In biological models for the disease, this leads to upregulated mRNA translation and as a consequence, deficits in synaptic architecture and plasticity. Preclinical studies revealed that pharmacological interventions restore those deficits, which are thought to mediate the FXS cognitive and behavioral symptoms. Here, we characterized the de novo rate of protein synthesis in patients with FXS and their relationship with clinical severity. We measured the rate of protein synthesis in fibroblasts derived from 32 individuals with FXS and from 17 controls as well as in fibroblasts and primary neurons of 27 Fmr1 KO mice and 20 controls. Here, we show that levels of protein synthesis are increased in fibroblasts of individuals with FXS and Fmr1 KO mice. However, this cellular phenotype displays a broad distribution and a proportion of fragile X individuals and Fmr1 KO mice do not show increased levels of protein synthesis, having measures in the normal range. Because the same Fmr1 KO animal measures in fibroblasts predict those in neurons we suggest the validity of this peripheral biomarker. Our study offers a potential explanation for the comprehensive drug development program undertaken thus far yielding negative results and suggests that a significant proportion, but not all individuals with FXS, may benefit from the reduction of excessive levels of protein synthesis.

Reprioritisation of Liver Export Protein Synthesis in Patients with Decompensated Alcoholic Liver Disease

Background: Decompensated alcoholic liver disease is associated with abnormalities in protein synthesis. The relationship of this to reprioritisation of hepatic export proteins and markers of the systemic inflammatory response is unclear. We examined the longitudinal relationship between albumin and fibrinogen synthetic rates and disease severity in decompensated alcoholic liver cirrhosis. Patients and Methods: Hepatic protein synthetic rates were measured in patients with decompensated Childs grade B or C alcohol-related cirrhosis using a validated technique, based on incorporation of deuterated phenylalanine into the plasma pool of albumin and fibrinogen. As a measure of liver export protein reprioritisation, we calculated the fibrinogen and albumin synthetic rates and the Acute Phase Protein Quotient (APPQ; the ratio of these rates). Serum CRP and fibrin D-Dimer were recorded. Measurements were at baseline and on clinical recovery at 4-6 weeks. Results: 17 patients were studied. All patients had hypoalbuminaemia with elevated median C-reactive protein (CRP), D-dimer, bilirubin and prothrombin times. Median albumin and fibrinogen synthetic rates were reduced resulting in marginally increased APPQ. On follow up (n=10), there was reduction in Child-Pugh score (p<0.01), plasma concentrations of Fibrin D-dimer (p<0.01), CRP (p<0.01), bilirubin (p<0.01) and prothrombin time (p<0.01). Plasma albumin concentrations increased (p<0.01) and synthetic rates of both albumin (p<0.05) and fibrinogen (p<0.10) increased marginally such that median APPQ remained similar. Conclusion: Patients with decompensated alcohol-related cirrhosis had low albumin and fibrinogen synthetic rates and raised CRP indicative of systemic inflammation. On recovery, albumin synthetic rate increased and CRP levels fell, although albumin and fibrinogen synthetic rates remained below normal. Further studies assessing the interaction between protein synthesis and systemic inflammation in chronic liver disease are indicated.

Habitual Myofibrillar Protein Synthesis Is Normal in Patients with Upper GI Cancer Cachexia.

Skeletal muscle wasting and weight loss are characteristic features of cancer cachexia and contribute to impaired function, increased morbidity, and poor tolerance of chemotherapy. This study used a novel technique to measure habitual myofibrillar protein synthesis in patients with cancer compared with healthy controls. An oral heavy water (87.5 g deuterium oxide) tracer was administered as a single dose. Serum samples were taken over the subsequent week followed by a quadriceps muscle biopsy. Deuterium enrichment was measured in body water, serum alanine, and alanine in the myofibrillar component of muscle using gas chromatography-pyrolysis-isotope ratio mass spectrometry and the protein synthesis rate calculated from the rate of tracer incorporation. Net change in muscle mass over the preceding 3 months was calculated from serial CT scans and allowed estimation of protein breakdown. Seven healthy volunteers, 6 weight-stable, and 7 weight-losing (≥5% weight loss) patients undergoing surgery for upper gastrointestinal cancer were recruited. Serial CT scans were available in 10 patients, who lost skeletal muscle mass preoperatively at a rate of 5.6%/100 days. Myofibrillar protein fractional synthetic rate was 0.058%, 0.061%, and 0.073%/hour in controls, weight-stable, and weight-losing patients, respectively. Weight-losing patients had higher synthetic rates than controls (P = 0.03). Contrary to previous studies, there was no evidence of suppression of myofibrillar protein synthesis in patients with cancer cachexia. Our finding implies a small increase in muscle breakdown may account for muscle wasting.

P4.42 Muscle protein synthesis in patients with Dystrophia Myotonica type 1

Dystrophia Myotonica type 1 (DM1) is the most common muscular dystrophy. Aerobic exercise improves fitness in DM1 patients. Whether the improvement is caused by improved cardio-respiratory fitness or increased muscle net-protein synthesis is not known. In healthy individuals exercise, especially in combination with protein-intake improved muscle protein anabolic state. Therefore, we investigated how exercise combined with protein-supplement affected skeletal muscle metabolism in DM1 patients. Seven DM1 patients and 6 healthy controls were studied during two trials with and without oral protein-supplementation for 3h in the recovery phase from exercise.

Effects of recombinant human growth hormone on enterocutaneous fistula patients

To explore the effects of recombinant human growth hormone (rhGH) on intestinal mucosal epithelial cell proliferation and nutritional status in patients with enterocutaneous fistula. Eight patients with enterocutaneous fistulas received recombinant human growth hormone (10 microg/d) for 7 d. Image analysis and immunohistochemical techniques were used to analyse the expression of proliferating cell nuclear antigen (PCNA) in intestinal mucosal epithelial cells in biopsy samples from the patients who had undergone an endoscopic biopsy through the fistula at day 0, 4 and 7. Body weights, nitrogen excretion, serum levels of total proteins, albumin, prealbumin, transferrin and fibronectin were measured at day 0, 4 and 7. Significant improvements occurred in the expression of PCNA in the intestinal mucosal epithelial cells at day 4 and 7 compared to day 0 (24.93 +/- 3.41%, 30.46 +/- 5.24% vs 12.92 +/- 4.20%, P < 0.01). These changes were accompanied by the significant improvement of villus height (500.54 +/- 53.79 microm, 459.03 +/- 88.98 microm vs 210.94 +/- 49.16 microm, P < 0.01), serum levels of total proteins (70.52 +/- 5.13 g/L, 74.89 +/- 5.16 g/L vs 63.51 +/- 2.47 g/L, P < 0.01), albumin (39.44 +/- 1.18 g/L, 42.39 +/- 1.68 g/L vs 35.74 +/- 1.75 g/L, P < 0.01) and fibronectin (236.3 +/- 16.5 mg/L, 275.8 +/- 16.9 mg/L vs 172.5 +/- 21.4 mg/L, P < 0.01) at day 4 and 7, and prealbumin (286.38 +/- 65.61 mg/L vs 180.88 +/- 48.28 mg/L, P < 0.05), transferrin (2.61 +/- 0.12 g/L vs 2.41 +/- 0.14 g/L, P < 0.05) at day 7. Nitrogen excretion was significantly decreased at day 7 (3.40 +/- 1.65 g/d vs 7.25 +/- 3.92 g/d, P < 0.05). No change was observed in the body weight. Recombinant human growth hormone could promote intestinal mucosal epithelial cell proliferation and protein synthesis in patients with enterocutaneous fistula.

認知症病棟で発生した紫色蓄尿バッグ症候群(PUBS）6例の生化学的および細菌学的検討

Purple urine bag syndrome is a condition in which the urinary catheter bag turns purple. A tryptophan-indigo hypothesis has been proposed as the mechanism of PUBS, in which bacterial decomposition of tryptophan in gut associated with chronic constipation, bacterial overgrowth in the urinary tract and alkaline urine causes production of indigo and discoloration. We considered that further investigation of cases was needed. We investigated 6 cases exhibiting PUBS (3 males and 3 females). All cases had chronic constipation. Oral ingestion was impossible in one case. PUBS disappeared after antibiotic treatment (3 cases) or spontaneously (one case). Alkaline urine and indicanuria were not found in all cases that showed the disappearance of PUBS. In bacterial culture of urine during the exhibition of PUBS, Enterococcus faecalis was isolated together with Morganella morganii (3 cases) and Pseudomonas aeruginosa (one case). Single infections by Klebsiella pneumoniae or Citrobacter species were also found. After disappearance of PUBS, infected bacterial species changed but no cases showed sterile urine. Urine and blood alpha-amino-n-butyric acid levels reduced after the disappearance of PUBS whereas tryptophan levels did not show related changes. In one case, blood protein concentration increased after the spontaneous disappearance of PUBS. Indicanuria and alkalization of urine from urinary catheter bag were more intense than of fresh urine. The present results generally support the 'Tryptophan-indigo hypothesis'. Furthermore, it was suggested that additional factors associated with the occurrence of PUBS are an environment that facilitates specific bacterial growth in a hospital as well as abnormal metabolism relating to alpha-amino-n-butyric acid and reduced protein synthesis in patients.

Amino acid metabolism and inflammatory burden in ovarian cancer patients undergoing intense oncological therapy

Cancer and oncological therapy are associated with a progressive physical deterioration, malnutrition, and enhanced inflammatory burden. Our considerable data showing the strong anabolic potential of amino acids (AA) led us to test whether AA can acutely stimulate muscle protein synthesis in cancer patients (CA) undergoing intense chemotherapy. Mixed muscle fractional synthesis rate (FSR), rates of phenylalanine appearance and disappearance (Ra and Rd), and net phenylalanine balance (NB) were measured during a primed constant infusion of L-[ring-(2)H(5)]phenylalanine. Blood and muscle tissue samples were collected in the basal state and following ingestion of 40 g of AA given in 30 mL boluses every 10 min for 3h. Serum and tissue cytokines and NF-kappaB expression in skeletal muscle were measured and compared to normative, healthy older controls (OC). Skeletal muscle TNF-alpha, IL-6, and NF-kappaB were elevated in CA. FSR and model-derived protein synthesis (Rd) increased significantly from basal to AA (FSR: 0.052+/-0.009 vs. 0.120+/-0.008%h(-1), P<0.001; Rd: 23.1+/-4.1 vs. 36.4+/-5.0 nmol min(-1) 100 mL leg(-1), P0.05). Model-derived protein breakdown (Ra) remained unchanged from basal to AA. Phenylalanine NB improved from a negative basal value (-16+/-2) to zero (0.8+/-6 nmol min(-1) 100 ml leg(-1), P0.05) following AA. Despite advanced cancer, ongoing therapy, and an enhanced inflammatory burden, AA were capable of acutely stimulating muscle protein synthesis in these patients.

Role of branched-chain amino acids in liver disease: the evidence for and against

There is ample evidence that patients with liver disease have an ongoing energy and protein catabolism. Nutritional management in these patients must receive high priority. The administration of branched-chain amino acids to patients with liver disease has been a controversial subject. This review is an update on the data available from various studies involving branched-chain amino acids supplementation in patients with chronic liver disease and associated complications. This review summarizes the results of nutritional interventions involving branched-chain amino acids supplementation carried out in different centres around the world. It is interesting to note that no toxic effects of branched-chain amino acids supplementation have been reported in any of these trials. Administration of branched-chain amino acids stimulates hepatic protein synthesis in patients with chronic liver disease and this could contribute significantly to improving their nutritional status, and result in a better quality of life. The beneficial role of branched-chain amino acids supplementation in patients with chronic hepatic encephalopathy has been clearly documented in some studies but the exact mechanism of action is still not clear.

Effect of an enteral diet supplemented with a specific blend of amino acid on plasma and muscle protein synthesis in ICU patients

Polytrauma patients are characterized by a negative nitrogen balance and muscle wasting. Standard nutrition is relatively inefficient to improve muscle protein turnover. The aim of this study was to investigate the effect of enteral nutrition (EN) supplemented with specific amino acids on protein metabolism in polytrauma patients. In a double blind study, 12 polytrauma patients were randomized to receive EN supplemented with either a mixture of cysteine, threonine, serine and aspartate (AA patients) or alanine at isonitrogenous levels (Ala patients). An intravenous infusion of l-[1-(13)C]-leucine was performed in the fed state between day 9 and 12 post-injury (Df) in patients and in a group of healthy volunteers (n=8) (EN+Ala) to measure whole body leucine kinetics, plasma and muscle protein synthesis rates. Nitrogen balance, 3-methyl histidine excretion were measured from day 3 to Df. The contribution of total plasma proteins to whole body protein synthesis was greatly increased, from 11% in healthy volunteers to about 25% in polytrauma patients. AA supplementation had no effect on nitrogen balance, leucine kinetics or plasma protein synthesis in patients. In contrast, the urinary excretion of 3-methyl histidine tended to decrease along the study in the AA supplemented group compared to an increase in the Ala group. Muscle protein synthesis tended to be higher in the AA group than in the Ala group (46%, P=0.065). During injury, an increased supply of cysteine, threonine, serine and aspartate could be able to better cover the specific amino requirements, thus resulting in improved muscle protein synthesis without impairment of acute phase protein synthesis.

The Effect of Hyperglycemic Hyperinsulinemia on Small‐Intestinal Mucosal Protein Synthesis in Patients After Surgical Stress

Hyperglycemic hyperinsulinemia cannot stimulate intestinal protein synthesis in healthy individuals but does so in conditions characterized by an altered somatotropic axis such as diabetes. Only in a state of growth hormone resistance (high growth hormone but low insulin like growth factor [IGF-1] concentrations), extra insulin may acutely reverse the impaired, growth-hormone-induced IGF-1 release, thereby exerting anabolic actions at the intestinal tract. Growth hormone resistance can be also found in patients after surgical stress. Therefore, we wanted to test the hypothesis whether hyperglycemic hyperinsulinemia would stimulate ileal protein synthesis in the latter condition. Mass spectrometry techniques (capillary gas chromatography/combustion isotope ratio mass spectrometry) were used to directly determine the incorporation rate of 1-[(13)C]-leucine into ileal mucosal protein. All subjects had an ileostomy, which allowed easy access to the ileal mucosa, and consecutive sampling from the same tissue was performed during continuous isotope infusion (0.16 mumol/kg min). Isotopic enrichments and fractional protein synthesis were determined at baseline (period I) and after a 4-hour glucose infusion (170 mg/kg/h) or after infusion of saline (control group) (period II). In controls, ileal protein synthesis declined significantly during prolonged isotope infusion (period I: 1.11 +/- 0.14%/h, period II: 0.39 +/- 0.13%/h, p < .01). In contrast, ileal protein synthesis remained constant during glucose infusion (period I: 1.32 +/- 0.35%/h, period II: 1.33 +/- 0.21%/h, n.s. vs period I, but p < .005 vs the corresponding value at the end of period II in the control group). Using the continuous tracer infusion technique, ileal protein synthesis seemingly declines over a short time in control subjects. We found evidence that this artificial decline was due to mass effects of a rapidly turning over mucosa protein pool in which an isotopic plateau was reached during the experiment and of which the size amounted to approximately 4% of the total mixed protein pool. Maintenance of ileal protein synthesis during glucose infusion therefore indicates a rise of ileal protein synthesis in a slowly turning over protein pool. This effect in postsurgical patients would be compatible with the concept of intestinal insulin action to depend on the specific clinical state (eg, growth hormone resistance).

Stimulation of protein synthesis in human tumours by parenteral nutrition: Evidence for modulation of tumour growth

Eighteen patients with localized colorectal carcinoma were randomized to receive intravenous nutrition or to be fasted during the 24 h before surgery. Protein synthesis, an index of tumour growth, was then measured by the incorporation of [13C]leucine into tumour protein immediately before surgery. The mean (s.e.m.) rate of tumour protein synthesis in patients receiving nutrition (42.7(3.5) per cent per day) was 89 per cent higher than the rate in the fasted group (22.6(1.9) per cent per day) (P = 0.002). As tumours consist of a variety of different cell types, in vitro rates of protein synthesis were measured in malignant cells isolated from colorectal tumours and cultured with autologous serum obtained from the patient in either the fasted or the fed state. There was a mean increase of 81 per cent in protein synthesis when fed rather than fasted serum was used (P less than 0.02), indicating that the malignant cells themselves respond to nutrient supply. This increase in tumour protein synthesis provides the first evidence in vivo that the exogenous supply of nutrients can modulate the rate of growth of a human tumour.

HIV-associated nephropathy: virologic issues related to renal sclerosis.

Human immunodeficiency virus (HIV) associated nephropathy (HIVAN) is the most common disease delineated in biopsy series of patients with HIV infection and renal disease. Although the renal histologic lesions in patients with HIV infection present a spectrum of findings, several groups have emphasized characteristic clinical and pathologic features of HIVAN. Consensus has since been reached that HIVAN has a distinctive pathology, consistent with focal segmental glomerulosclerosis, but involving all subunits of the kidney. Several studies have linked the pathogenesis of focal glomerulosclerosis to abnormalities of the glomerular epithelial cell. Recent advances in molecular histologic studies and treatment link the pathogenesis of HIVAN to factors associated with the viral lifecycle. Cytopathic effects of HIV gene products, apoptosis mediated by HIV infection, the elaboration of chemokines and cytokines as a result of viral or host protein synthesis in patients with genetic susceptibility to nephropathy, and the subversion of the host metabolic and synthetic machinery by the virus might be sufficient to create a rapidly progressive disease. If HIV infects and has cytopathic effects on glomerular epithelial cells, and dysfunction of these cells is intimately related to the pathogenesis and progression of renal disease, a reasonable pathogenic mechanism for the development of HIVAN may be inferred. The similarities of HIVAN and the collapsing variant of focal segmental glomerulosclerosis pose the intriguing possibility that the latter is a viral illness as well. The disease is marked by glomerular sclerosis with varying degrees of collapse, tubular epithelial cell degeneration, simplification, microcystic dilatation, interstitial fibrosis and immune cell infiltration. At the level of electron microscopy, tubular reticular inclusions in renal endothelial cells are a typical, but not pathognomonic feature of HIVAN.

Effect of tumor removal on mucosal protein synthesis in patients with colorectal cancer.

It is currently controversial whether mucosal hyperproliferation is involved in colorectal cancerogenesis. The purpose of the present study was to examine protein synthetic rate as an indicator of potential tissue proliferation in grossly normal rectal mucosa from cancer-bearing subjects and to compare this rate with that in mucosa from subjects posttumor removal. Six postabsorptive patients with localized rectal cancer and five postsurgical control subjects received a primed constant infusion of [1-(13)C]leucine (0.16 micromol/kg min, 9.6 micromol/kg prime). Forceps biopsies from the mucosa were taken after 3 and 6 h. Protein synthesis was calculated from protein-bound leucine enrichment (determined by capillary GC-combustion IRMS) and from the enrichment of free intracellular leucine (determined by GC-quadrupole MS). In cancer-bearing subjects, mucosal protein synthesis amounted to 1.28 +/- 0.24%/h. This rate was significantly higher (P < 0.05) than the corresponding rate of mucosa from patients after cancer removal (0.69 +/- 0.09%/h). These findings do not support the concept that colorectal cancer originates from a proliferative disease of the whole colon. Increased mucosal protein synthesis appears to depend on the presence of the tumor itself and should therefore be considered a secondary phenomenon.

Plasma protein synthesis in patients with low-grade nephrotic proteinuria.

Overt nephrotic syndrome is characterized by albumin and fibrinogen hyperproduction and reduced very low density lipoprotein apolipoprotein B-100 (VLDL apoB-100) clearance. Whether similar changes also occur in low-grade proteinuria is not known. Thus we measured albumin, fibrinogen, and VLDL apoB-100 kinetics in six patients with modest proteinuria and normal creatinine clearance (P) and in ten control subjects (C) by leucine tracer infusion and precursor-product relationships. In P, plasma albumin concentration was decreased (P < 0.003), whereas concentrations of fibrinogen and VLDL apoB-100 were increased (P < 0.001). In P, albumin fractional secretion rate (FSR) was increased (P < 0.01), fibrinogen FSR was normal, and VLDL apoB-100 FSR was decreased (P < 0.03). As a result, in P, absolute secretion rates (ASR) of albumin and fibrinogen were increased (P < 0.03), whereas VLDL apoB-100 ASR was normal. Albumin FSR was inversely correlated to oncotic pressure in P but not in C. These findings suggest that low-grade nephrotic proteinuria is characterized by simultaneous multiple alterations in turnover rates of albumin, fibrinogen, and VLDL apoB-100. Their pathogenesis, however, appears to be multifactorial.

The effect of insulin on human small intestinal mucosal protein synthesis

Background & Aims: Insulin deficiency was recently shown to stimulate splanchnic protein synthesis in vivo, whereas insulin enhances small intestinal mucosal cell proliferation in vitro. Because insulin is a postprandial hormone, it was hypothesized that it has an important role in regulating small intestinal protein synthesis in humans. Methods: Small intestinal mucosal protein synthesis was measured in C-peptide–negative patients with type 1 diabetes mellitus during insulin deprivation (n = 6) and during insulin treatment (n = 6) and in nondiabetic control subjects (n = 6). Mucosal protein synthesis was measured from the increment of [ 13C]leucine enrichment in endoscopically obtained duodenal mucosa samples during a primed continuous infusion of L-[1- 13C]leucine. Results: During insulin treatment, the rate of mucosal protein synthesis in patients with type 1 diabetes was similar (1.32% ± 0.05%/h) to that of nondiabetic controls (1.33% ± 0.06%/h). However, during insulin deprivation, the mucosal protein synthesis rate in patients with type 1 diabetes was significantly lower (1.15% ± 0.33%/h) than during either insulin treatment ( P = 0.01) or in nondiabetic controls ( P = 0.04). Conclusions: These studies show that insulin is required for the maintenance of normal rates of protein synthesis in small intestinal mucosa. Because protein synthesis is an essential component of the remodeling process of this fast turning over tissue, the decline in the synthesis rate of small intestinal mucosa during insulin deprivation may be a contributing factor in the development of gastrointestinal complications that occur in poorly controlled type 1 diabetic patients. GASTROENTEROLOGY 2000;118:299-306

Threonine and Methionine Are Limiting Amino Acids for Protein Synthesis in Patients with AIDS

This study was conducted to identify the most rate-limiting amino acids for whole-body protein synthesis in acquired immunodeficiency syndrome (AIDS) patients. We postulated that an essential amino acid that would be rate limiting in AIDS should have a low basal plasma concentration and should remain at a low level during amino acid infusion. Seven male AIDS patients (median age 37 y, CD4 cell count: 76 mm−3) without any clinically active opportunistic infection during the month before the experiment were infused intravenously with a complete amino acid-glucose mixture for 2.5 h. Eight healthy volunteers were used as controls. Before the infusion, the concentrations of most free essential amino acids (methionine, threonine, histidine, isoleucine, leucine and tryptophan) were significantly lower (P < 0.05) in AIDS patients than in controls. Most plasma free essential amino acids increased significantly during infusion. However, the absolute increase above basal levels for threonine, valine, lysine, (P < 0.05) and methionine (P < 0.073) was smaller in AIDS patients than in control subjects. Thus, threonine and possibly methionine may be rate limiting for whole-body protein synthesis in AIDS patients, suggesting that there are selective amino acid requirements in patients with AIDS.

Measurement of duodenal mucosal protein synthesis in patients with coeliac disease: [formula omitted], S Downie∗, M J Rennie∗, R I Russell. Department of Gastroenterology, Royal Infirmary, Glasgow and Department of Anatomy & Physiology∗, University of Dundee, Dundee, Scotland

Measurement of duodenal mucosal protein synthesis in patients with coeliac disease: [formula omitted], S Downie∗, M J Rennie∗, R I Russell. Department of Gastroenterology, Royal Infirmary, Glasgow and Department of Anatomy & Physiology∗, University of Dundee, Dundee, Scotland