Blood Methionine Research Articles

Distribution of methionine between cells and incubation medium in suspension of rat hepatocytes

Methionine is an essential amino acid involved in many significant intracellular processes. Aberrations in methionine metabolism are associated with a number of complex pathologies. Liver plays a key role in regulation of blood methionine level. Investigation of methionine distribution between hepatocytes and medium is crucial for understanding the mechanisms of this regulation. For the first time, we analyzed the distribution of methionine between hepatocytes and incubation medium using direct measurements of methionine concentrations. Our results revealed a fast and reversible transport of methionine through the cell membrane that provides almost uniform distribution of methionine between hepatocytes and incubation medium. The steady-state ratio between intracellular and extracellular methionine concentrations was established within a few minutes. This ratio was found to be 1.06±0.38, 0.89±0.26, 0.67±0.16 and 0.82±0.06 at methionine concentrations in the medium of 64±19, 152±39, 413±55, and 1,204±104 μmol/L, respectively. The fast and uniform distribution of methionine between hepatocytes and extracellular compartments provides a possibility for effective regulation of blood methionine levels due to methionine metabolism in hepatocytes.

Short communication: Transport of 2-hydroxy-4-methyl-thio-butanoic isopropyl ester by rumen epithelium in vitro

Our objective was to evaluate the potential of rumen epithelium to transport 2-hydroxy-4-(methylthio)-butanoic isopropyl ester (HMBi) using the Ussing chamber technique. Rumen tissues were obtained from a nearby slaughterhouse, separated from the muscle and serosal layer as quickly as possible after exsanguination, placed in buffer, and gassed with 95:5 (vol/vol) O2:CO2 before tissue mounting. Two levels of HMBi (0.44 and 0.88mg/mL) and 2 incubation times (120 and 180min) were used in 12 chambers with 3 replicates per treatment with an exposed surface area of 2cm2. Four separate experiments were conducted (n=16). Concentrations of HMBi and methionine hydroxy analog (HMB) were measured by HPLC in rumen-side and serosal-side buffers. Data are expressed as percentage of added HMBi. Initial time samples were taken for comparison with incubated samples. Adding the HMBi-buffer mixture to the rumen side caused an immediate release of HMB (mean=6.3%). Breakdown of HMBi to HMB at initial time was due to hydrolysis reactions at the epithelial surface. Overall, a small and variable amount of HMBi was transferred to the serosal buffer (mean of 0.58% across both times and both concentrations). A larger amount of HMB (8.94%) was isolated in the serosal buffer. Increasing incubation time increased the amount of HMB in the ruminal buffer (34.0% at 120min vs. 43.4% at 180min) and decreased the amount of HMBi (37.9% at 120min vs. 28.1% at 180min). These data indicate that very limited amounts of HMBi may cross the rumen epithelium. The amount of HMB isolated on the serosal side was about 10 times higher than HMBi. The hydrolysis of HMBi to HMB required the presence of rumen tissue or perhaps microbes attached to the tissue. Based on this in vitro system, direct transport from the rumen would not explain rapid blood methionine increases observed when HMBi is fed.

An Allosteric Mechanism for Switching between Parallel Tracks in Mammalian Sulfur Metabolism

Methionine (Met) is an essential amino acid that is needed for the synthesis of S-adenosylmethionine (AdoMet), the major biological methylating agent. Methionine used for AdoMet synthesis can be replenished via remethylation of homocysteine. Alternatively, homocysteine can be converted to cysteine via the transsulfuration pathway. Aberrations in methionine metabolism are associated with a number of complex diseases, including cancer, anemia, and neurodegenerative diseases. The concentration of methionine in blood and in organs is tightly regulated. Liver plays a key role in buffering blood methionine levels, and an interesting feature of its metabolism is that parallel tracks exist for the synthesis and utilization of AdoMet. To elucidate the molecular mechanism that controls metabolic fluxes in liver methionine metabolism, we have studied the dependencies of AdoMet concentration and methionine consumption rate on methionine concentration in native murine hepatocytes at physiologically relevant concentrations (40–400 µM). We find that both [AdoMet] and methionine consumption rates do not change gradually with an increase in [Met] but rise sharply (∼10-fold) in the narrow Met interval from 50 to 100 µM. Analysis of our experimental data using a mathematical model reveals that the sharp increase in [AdoMet] and the methionine consumption rate observed within the trigger zone are associated with metabolic switching from methionine conservation to disposal, regulated allosterically by switching between parallel pathways. This regulatory switch is triggered by [Met] and provides a mechanism for stabilization of methionine levels in blood over wide variations in dietary methionine intake.

Subacute combined degeneration of the spinal cord in cblC disorder despite treatment with B12

Subacute combined degeneration (SCD) of the spinal cord is a characteristic complication of vitamin B12 deficiency, but it has never been neuropathologically demonstrated in a B12-inborn error of metabolism. In this report SCD is documented in a 15-year-old boy with early-onset cobalamin C (cblC) disorder. The neuropathologic findings included multifocal demyelination and vacuolation with predilection for the dorsal and lateral columns at the mid-thoracic level of the spinal cord, confirming the similarity of SCD in cblC disorder to the classic adult SCD due to vitamin B12 deficiency. SCD developed in this boy despite treatment for cblC disorder that began at 3 months of age. There is clinical and experimental evidence to suggest that a deficiency in remethylation with concomitant reduction in brain methionine may be the cause of SCD. In this patient plasma methionine levels were low without betaine and/or l-methionine supplementation and in the normal range for only a 2-year period during compliance with therapy. In cblC disorder, a consistent increase in blood methionine to high normal or above normal levels by the use of betaine and l-methionine supplementation may be helpful in preventing SCD. This is especially important now that the presymptomatic detection of cblC disorder is possible through the expansion of newborn screening.

Proton MR spectroscopy in hyperhomocysteinemia with elevated blood methionine levels

Proton magnetic resonance spectroscopy imaging (MRSI) and serial MRI were performed on a 10-year-old girl with B6 unresponsive cystathionine B-synthase (CBS) deficiency who developed high methionine levels while on betaine therapy. At presentation, T2-weighted sequences showed diffuse white matter (WM) hyperintensity and sulcal effacement, while MRSI metabolite concentrations were normal. Four months later, after the betaine therapy was discontinued and a methionine-restricted diet with vitamin B6, B12, and folate supplementation was initiated, blood methionine levels and MRI findings returned to normal. Normal MRSI at presentation was predictive of a positive outcome despite the markedly abnormal initial MRI results.

Neonatal Screening by Tandem Mass Spectrometry

After completing this article, readers should be able to: 1. List the major categories of diseases detectable by expanded newborn screening. 2. Give examples of the benefits and limitations of screening by tandem mass spectrometry. 3. Describe the appropriate follow-up for an abnormal screening result. The screening of newborns for inherited metabolic disorders is a well-established public health activity, first implemented in the early 1960s for the presymptomatic identification of patients who have phenylketonuria (PKU). Since then, significant technological advances, most importantly the development of tandem mass spectrometry (MS/MS), have enabled the detection of an increasing number of metabolic disorders in newborn blood. This article reviews the basic technology of MS/MS and its application to newborn screening, with the aim of highlighting the benefits and pointing out some limitations of this powerful technology. Early screening for PKU used the bacterial inhibition assay of Guthrie, in which inhibition of bacterial growth on an agar medium, introduced by the addition of a phenylalanine analog, was overcome by the presence of dried blood discs containing high levels of phenylalanine (such as from a PKU patient). This approach later was adapted for the detection of leucine in patients who have maple syrup urine disease (MSUD), methionine in patients who have classic homocystinuria, and tyrosine in patients who have tyrosinemia. Over the years, other techniques have been implemented to screen for additional disorders, including congenital hypothyroidism, biotinidase deficiency, galactosemia, hemoglobinopathies, and congenital adrenal hyperplasia. The classic criteria used to determine whether a disorder is suitable for screening include: 1) the disorder is clinically and biochemically well defined, 2) it is associated with significant morbidity or mortality, 3) an effective treatment is available, and 4) there is a simple and safe screening test. (1) By the early 1990s, all states offered screening for at least PKU and congenital hypothyroidism, …

Regulation of Mammalian Liver Methionine Adenosyltransferase

S-adenosylmethionine (SAM) is an essential metabolite in all cells. SAM is the most important biological methyl group donor and is a precursor in the synthesis of polyamines. Methionine adenosyltransferase (MAT; EC 2.5.1.6) catalyzes the only known SAM biosynthetic reaction from methionine and ATP. In mammalian tissues, three different forms of MAT (MAT I, MAT III and MAT II) have been identified that are the product of two different genes (MAT1A and MAT2A). Although MAT2A is expressed in all mammalian tissues, the expression of MAT1A is primarily restricted to adult liver. In mammals, up to 85% of all methylation reactions and as much as 48% of methionine metabolism occurs in the liver, which indicates the important role of this organ in the regulation of blood methionine. Recent evidence indicates that not only is SAM the main biological methyl group donor and an intermediate metabolite in methionine catabolism, but it is also an intracellular control switch that regulates essential hepatic functions such as liver regeneration and differentiation as well as the sensitivity of this organ to injury. Therefore, knowledge of factors that regulate the activity of MAT I/III, the specific liver enzyme, is essential to understand how cellular SAM levels are controlled.

S‐Adenosylmethionine: a control switch that regulates liver function

Genome sequence analysis reveals that all organisms synthesize S-adenosylmethionine (AdoMet) and that a large fraction of all genes is AdoMet-dependent methyltransferases. AdoMet-dependent methylation has been shown to be central to many biological processes. Up to 85% of all methylation reactions and as much as 48% of methionine metabolism occur in the liver, which indicates the crucial importance of this organ in the regulation of blood methionine. Of the two mammalian genes (MAT1A, MAT2A) that encode methionine adenosyltransferase (MAT, the enzyme that makes AdoMet), MAT1A is specifically expressed in adult liver. It now appears that growth factors, cytokines, and hormones regulate liver MAT mRNA levels and enzyme activity and that AdoMet should not be viewed only as an intermediate metabolite in methionine catabolism, but also as an intracellular control switch that regulates essential hepatic functions such as regeneration, differentiation, and the sensitivity of this organ to injury. The aim of this review is to integrate these recent findings linking AdoMet with liver growth, differentiation, and injury into a comprehensive model. With the availability of AdoMet as a nutritional supplement and evidence of its beneficial role in various liver diseases, this review offers insight into its mechanism of action.

Reduction of false negative results in screening of newborns for homocystinuria.

Mental retardation and other disabilities (including ectopia lentis, osteoporosis, and thromboembolism) in patients who have homocystinuria as a result of a deficiency of cystathionine beta-synthase can be prevented by the screening of newborns with measurement of blood methionine, followed by the early treatment of affected infants. Many infants with this disorder, however, are not identified by screening and have irreversible brain damage. We reviewed the results of neonatal screening for homocystinuria over a period of 32 years in New England. Additional specimens were requested for repeated analysis when blood methionine measurements were at or above the established cutoff level. Homocystinuria due to cystathionine beta-synthase deficiency was confirmed by quantitative amino acid analyses. For the first 23.5 years of the review period, the blood methionine cutoff value was 2 mg per deciliter (134 micromol per liter). Among the 2.2 million infants screened during that period, 8 with homocystinuria were identified (1:275,000). In 1990, the cutoff value was reduced to 1 mg per deciliter (67 micromol per liter). Among the 1.1 million infants screened in the subsequent 8.5 years, 7 with the disorder were identified (1:157,000). During the latter period, the specimens were collected from six of the seven infants when they were two days of age or less; five of the six had blood methionine concentrations below 2 mg per deciliter. Use of the reduced cutoff level increased the false positive rate from 0.006 percent to 0.03 percent. A cutoff level for blood methionine of 1 mg per deciliter in neonatal screening tests for homocystinuria should identify affected infants who have only slightly elevated concentrations of methionine and reduce the frequency of false negative results.

Effects of dietary cysteine on blood sulfur amino acid, glutathione, and malondialdehyde concentrations in cats

Abstract Objective To determine effects of dietary cysteine on blood sulfur amino acids (SAA), reduced glutathione (GSH), oxidized glutathione (GSSG), and malondialdehyde (MDA) concentrations in cats. Animals 12 healthy adult cats. Procedure Cats were fed diets with a nominal (0.50 g/100 g dry matter [DM]), moderate (1.00 g/100 g DM), or high (1.50 g/100 g DM) cysteine content in a 3 × 3 Latin square design with blocks of 8 weeks’ duration. Venous blood samples were collected after each diet had been fed for 4 and 8 weeks, and a CBC and serum biochemical analyses were performed; poikilocyte, reticulocyte, and Heinz body counts were determined; and MDA, GSH, GSSG, and SAA concentrations were measured. Results Blood cysteine and MDA concentrations were not significantly affected by dietary cysteine content. Blood methionine, homocysteine, and GSSG concentrations were significantly increased when cats consumed the high cysteine content diet but not when they consumed the moderate cysteine content diet, compared with concentrations obtained when cats consumed the nominal cysteine content diet. Blood GSH concentrations were significantly increased when cats consumed the moderate or high cysteine content diet. Conclusions Increased dietary cysteine content promotes higher blood methionine, homocysteine, GSH, and GSSG concentrations in healthy cats. Clinical Relevance Supplemental dietary cysteine may be indicated to promote glutathione synthesis and ameliorate adverse effects of oxidative damage induced by disease or drugs. (Am J Vet Res 1999;60:328–333)

Inadequate Laboratory Technique for Amino Acid Analysis Resulting in Missed Diagnoses of Homocystinuria

The diagnosis of cystathionine β-synthase (CBS) deficiency and disorders of methionine remethylation depend on the demonstration of homocyst(e)inemia and homocystinuria. To be detectable in the usual analysis of amino acids, homocystine must remain as a free disulfide in the plasma or serum. However, homocystine undergoes an exchange reaction with free sulfhydryls, including cysteine, readily forming covalent disulfide bonds with these thiols, and gradually disappears as a free amino acid (1). Because preparation for quantitative amino acid analysis usually includes deproteinization, protein-bound homocysteine is not detected by standard methods (2)(3)(4). Homocysteine-cysteine mixed disulfide, also increased in the homocystinurias, undergoes a similar exchange. This phenomenon is still not appreciated by many laboratories that perform amino acid analyses, and lack of awareness of the phenomenon continues to result in missing the diagnosis of homocystinuria. This was brought to our attention by the case of an infant with CBS-deficient homocystinuria. Routine newborn screening had identified an increased concentration of blood methionine (5). Quantitative plasma amino acid analysis 1 week later disclosed isolated hypermethioninemia. …

Newborn screening for homocystinuria: Irish and world experience.

Newborn screening for cystathionine beta-synthase deficiency (homocystinuria; HCU) was started in the late 1960s using a bacterial inhibition assay (BIA). At least seven countries have either national or regional screening programmes; 12 programmes are known to have discontinued. The worldwide incidence of HCU is approximately 1 in 335,000 but varies from 1:65,000 (Ireland) to 1:900,000 (Japan). Methodologies include the BIA, one-dimensional or thin-layer amino acid chromatography and, more recently, tandem mass spectrometry. The BIA diagnostic cut off concentration of blood methionine varies from 67 to 270 micromol/ (10-40 mg/l) with a median of 135 micromol/l (20 mg/l). In Ireland, 25 cases of HCU from 19 families have been identified from 1.58 million newborn infants since 1971; 21 cases were detected through the screening programme. Of the four missed cases, three were breast-fed at the time of blood collection and one was pyridoxine responsive. These findings were in broad agreement with the results from five other programmes, in which approximately one in every five cases was missed by the screening programme. Early hospital discharge, low protein intake, high blood methionine cut-off concentration and pyridoxine responsiveness were all identified as contributing to missed cases.

Intracarotid recombinant human tumor necrosis factor-alpha reduces cerebral blood flow and methionine uptake in rat brain tumors.

The aim of the present study is to understand the therapeutic effects of recombinant human tumor necrosis factor-alpha (rH-TNF) on hemocirculation and metabolism of brain tumors. Using double-label autoradiographic technique, we have monitored changes in regional cerebral blood flow (rCBF) and protein-bound fraction of (3H-methyl)-L-methionine, expressed as acid-insoluble fraction (AIF), in rat brain tumors following treatment with intracarotid rH-TNF. The central portion of tumors showed a significant decrease in rCBF and AIF at 4 hours after the injection (p < 0.01, p < 0.05, respectively, as compared with non-treated control rats), turned microscopically necrotic at 24 hours, and became more extensively necrotic at 72 hours. Tumor cells remained viable only in the peripheral portion of the tumors after the treatment. The peripheral portion also showed a moderate decrease in rCBF, but less change in AIF to 4 to 72 hours after the treatment. Neither ipsilateral nor contralateral non-involved cortex demonstrated appreciable changes in rCBF and AIF during the observed period. Intracarotid rH-TNF selectively reduces tumor rCBF and AIF, resulting in histological modification.

The importance of transmethylation reactions to methionine metabolism in sheep: effects of supplementation with creatine and choline.

The influence of administering the methylated products choline and creatine on methionine irreversible-loss rate (ILR) and recycling from homocysteine has been investigated in sheep fed close to energy and N equilibrium. Two methods to estimate methionine recycling were compared. The first involved [U-13C]methionine infused as part of a labelled amino acid mixture obtained from hydrolysed algal protein. In this approach the isotope dilution of methionine with all five C atoms labelled (m + 5) will represent the ILR which does not recycle through homocysteine, while that which includes molecules with C-1-C-4 labelled will allow for loss of the labelled methyl (5)-C atom and replacement by an unlabelled moiety in the remethylation of homocysteine. The second method involved a combined infusion of [1-13C]- and [S-methyl-2H3]methionine. These two approaches gave similar data for methionine ILR which does not include label recycled to the amino acid from homocysteine but differed for recycled methionine fluxes. Consequently the two procedures differed in the calculated extent of homocysteine methylation under control conditions (6 v. 28%). These extents of remethylation are within the range observed for the fed human subject, despite the fact that fewer dietary methyl groups are available for the ruminant. Using combined data from the infusions, significant depression of methionine recycling occurred in blood (P < 0.05), with a similar trend for plasma (P = 0.077), when choline plus creatine were infused. Wool growth, assessed by intradermal injection of [35S]cysteine, was not altered by supplementation with the methylated products. From changes in the label pattern of free methionine in aortal, hepatic portal and hepatic venous blood during U-13C-labelled algal hydrolysate infusion, the major sites of homocysteine remethylation appear to be the portal-drained viscera and the liver. This was confirmed by analysis of free methionine enrichments in various tissues following dual infusion of [1-13C]- and [S-methyl-2H3]methionine, with the greatest activities occurring in rumen, jejunum and liver. Of the non-splanchnic tissues examined, only kidney exhibited substantial methionine cycling; none was detected in muscle, heart, lung and skin. The implications of methyl group provision under net production conditions are discussed.

Temporal changes in concentrations of amino acids in plasma and whole blood of healthy neonatal foals from birth to two days of age

Summary Temporal changes, as well as differences in distribution, in concentrations of 24 amino acids in plasma and whole blood of neonatal foals were determined from birth to 2 days of age. In addition, differences in concentrations of amino acids in plasma between mare and foal pairs were determined at birth. Significant (P &lt; 0.05) hypoaminoacidemia existed for 15 amino acids in plasma of foals at birth, compared with mares (paired t-test). Concentrations of 7 amino acids (aspartate, glutamate, glutamine, glycine, hydroxyproline, phenylalanine, proline) in plasma of foals were higher (P &lt; 0.05) at birth than in mares, and concentrations of 2 (taurine, tryptophan) were not different (P &gt; 0.05). Significant (P &lt; 0.05) temporal changes for concentrations of 19 of 24 amino acids in plasma were observed during the 48-hour period. Concentrations of 13 of the 19 amino acids in plasma that had significant changes were higher (P &lt; 0.05) at 48 hours. Significant (P &gt; 0.05) effect of time on concentration of 5 amino acids (alanine, methionine, phenylalanine, taurine, threonine) in plasma was not found after birth. Temporal changes in concentrations of 7 amino acids (alanine, asparagine, glutamine, histidine, hydroxyproline, methionine, and threonine) in whole blood were not significantly (P &gt; 0.05) different from those in plasma. Temporal changes for concentrations of the remaining 17 amino acids in whole blood were significantly (P &lt; 0.05) different, compared with plasma. Distribution of the concentrations of 18 amino acids between whole blood and plasma was significantly (P &lt; 0.05) different. Concentrations of 5 amino acids (citrulline, cystine, glutamine, methionine, tryptophan) were significantly (P &lt; 0.05) lower in whole blood than in plasma, whereas concentrations of 13 amino acids were significantly (P &lt; 0.05) higher in whole blood vs plasma. Concentrations of 6 amino acids (asparagine, isoleucine, leucine, proline, serine, valine) in whole blood were not significantly different from concentrations in plasma. Significant differences in temporal patterns of concentrations of amino adds in plasma and whole blood may be attributable to nutritional or physiologic changes associated with parturition. Significant differences between concentrations of amino acids in whole blood and plasma may be attributable to ontogeny or specificity of transport systems across cell membranes.

Short Term Effect of Nitrous Oxide on Methionine and S-Adenosyl Methionine Concentrations

Concentrations of methionine and S-adenosyl methionine (SAM) have been measured in various tissues of rats exposed to 50% nitrous oxide for periods up to 80 min, a time by which there is detectable interference with thymidine synthesis in bone marrow. There were no significant changes of either SAM or methionine in plasma, whole blood or brain. Hepatic SAM was unchanged, but there was a highly significant reduction in hepatic methionine, to 62% of control, after 80 min exposure to nitrous oxide.

Nutritional therapy for selected inborn errors of metabolism.

Nutritional approaches are available for the management of several different classes of inborn metabolism errors. In phenylketonuria (PKU), phenylalanine is not properly metabolized; and its accumulation leads to neurologic dysfunction and metal retardation. Altering the diet to limit phenylalanine intake led to remarkable improvement in children with PKU. It was later found that instituting dietary therapy immediately after identification of the disorder in newborns prevented mental retardation. Throughout the 1960s nutritional therapies were found for other inborn disorders, including galactosemia, maple syrup urine disease, and homocystinuria. For the group of disorders associated with defects in the urea cycle, leading to profound hyperammonemia, therapy based on the concept of waste nitrogen excretion (i.e., by increasing excretion of urea cycle intermediates in the urine, nitrogen that would otherwise recycle as ammonia can be eliminated) dramatically produced better control of hyperammonemia and its consequences. Some inborn errors of metabolism respond to vitamin therapy. Biotin-related multiple carboxylase synthetase deficiency can be produced by either of two enzyme defects--holocarboxylase synthetase deficiency or biotinidase deficiency. Both are treatable with biotin supplementation. The symptoms of multiple carboxylase deficiency can also occur after intestinal resection or ingestion of raw eggs. Multiple carboxylase deficiency has been treated successfully in utero by giving the mother biotin supplements. Peroxisomal disorders may respond to dietary management. Liver disease in hereditary tyrosinemia may be accentuated by hypermethioninemia and treated by controlling the blood methionine level. Glycogen storage disease Type I, which causes hypoglycemia, can be controlled by oral administration of cornstarch.

SHORT TERM EFFECT OF NITROUS OXIDE ON METHIONINE AND S-ADENOSYL METHIONINE CONCENTRATIONS

Concentrations of methionine and S-adenosyl methionine (SAM) have been measured in various tissues of rats exposed to 50% nitrous oxide for periods up to 80 min, a time by which there is detectable interference with thymidine synthesis in bone marrow. There were no significant changes of either SAM or methionine in plasma, whole blood or brain. Hepatic SAM was unchanged, but there was a highly significant reduction in hepatic methionine, to 62% of control, after 80 min exposure to nitrous oxide.

Studies on fate of methionine in mice.

血液中のメチオニン濃度に近いトレーサーの標識メチオニン(Met)水溶液の微量を，C57BLマウスの尾静脈内(i．v．)に注射した．メチオニンの体内での代謝による分解の割合は，定められた標識位置の〔1-14C〕-Met，〔m-14C〕-Metおよび〔35S〕-Metを用いることによって求められた．i・v・投与後のkineticsとdynamicsについて詳細な資料を得た結果，下記のことが明らかとなった．(1)投与後5分で，大部分の標識体は器官・組織内へ取り込まれ，一部は最終代謝物14CO2となって，呼気中に回収された．血液中放射能濃度の最小を示す30分値は，呼気14CO2排泄の最大と一致した．30分以降の血液中放射能濃度の緩徐な上昇は，代謝物の一部血中への寄与によるものと思われた．(2)取り込まれたメチオニンは，少なくとも48時間までに40%強は体内で利用され〔1-14C〕，〔m-14C〕，〔35S〕の比を保っていた．(3)残りの60%のうち，40%は投与後6時間までに代謝を受け，〔1-14C〕部は主として呼気へ，〔m-14C〕部は呼気と尿へ，また，〔35S〕部は主として尿と一部糞へ排泄された．(4)最:後の20%の一部については，膵液，腸液中の酵素蛋白へ取り込まれて，48時間以内に消費分解されてしまうことが，ミクロオートラジオグラフィーによって明らかにされた．

Regional coupling of blood flow and methionine uptake in an experimental tumor assessed with autoradiography

Regional distribution of L-[methyl-14C] methionine (14C-MET) and 4-[18F] fluoro-antipyrine was compared using experimental rat tumors (AH109A) and a computerized autoradiogram image processor. Tissue distributions of the two tracers were found to be inhomogeneous in the tumor with nearly identical image patterns. Analysis of tissue radioactivities revealed that 82% of 14C-MET was derived from the acid insoluble fraction at 60 min after injection. The present study showed that 14C-MET uptake closely relates to tissue blood flow and may depend on its blood to tissue transport. Rapid incorporation of MET in the acid insoluble fraction implies that it is rapidly metabolized after transport into tumor tissue.