Metabolic alterations in adipose tissue during the early phase of experimental endotoxemia in humans

Adipose tissue plays an important role in energy homeostasis; however, there is only little knowledge about its metabolic activity during critical illness or sepsis. We assessed adipose tissue metabolic activity and local blood flow during experimental endotoxemia in otherwise healthy humans. In a prospective, placebo controlled and randomized experiment we measured changes in lactate, glycerol, and pyruvate concentrations in microdialysate samples of femoral adipose tissue after an intravenous bolus of lipopolysaccharide (LPS, 4 ng/kg). Intravenous endotoxin caused an early and constant increase in interstitial pyruvate, while formation of lactate in adipose tissue was not affected. In contrast, lactate levels in serum were elevated significantly after 90 min (p<0.05) and likewise, serum glycerol concentrations rose 90 min after LPS treatment (p<0.05) and 60 min earlier than in adipose tissue. Subcutaneous adipose tissue blood perfusion increased 2-fold while there was a strong decline in skin blood flow. Pyruvate accumulation in subcutaneous adipose tissue is an early marker of endotoxemia. While adipose tissue is a major source of serum glycerol and lactate in humans during physiological conditions, it contributes only little to increased serum lactate and glycerol levels during endotoxemia.     

Lactate interferes with ATP release from red blood cells

Upon exposure to low Po(2), the red blood cells of most species, including humans, release increased amounts of ATP that ultimately serves as a regulator of vascular tone matching oxygen supply with demand. In pathological conditions such as malaria and sepsis, a maldistribution of perfusion exists with its severity often correlated with the extent of elevation of serum lactate frequently in the absence of an alteration in pH. We hypothesized that the increased levels of lactate might impair the ability of red blood cells to appropriately respond to conditions of low Po(2), thus preventing its important blood flow regulatory role. Using an in vitro system and rabbit red blood cells, we evaluated the capacity of cells incubated with lactate to release increased amounts of ATP in response to acute exposure to low Po(2). We found that in the presence of lactate, the red blood cells did not release ATP. However, when sodium dichloroacetate, a drug used clinically to lower blood lactate levels, was added, ATP release was restored to levels that were not different from that of control cells (no lactate), even though intracellular levels of ATP were not. These results support the presence of a distinct flow regulatory pool of ATP within the red blood cell that can be independently regulated, and that lactate interferes with the ATP production within this pool, thereby diminishing the amount of ATP available for release on exposure to low Po(2). Therefore, if lactate levels can be reduced, the vascular regulatory capacity of the red blood cell should be restored, thus enabling the appropriate matching of oxygen supply with oxygen demand.     

体外循环超滤法影响小儿心内直视术患者血糖和血乳酸水平的研究

目的本研究旨在探讨体外循环超滤法对小儿心内直视术患者血糖和血乳酸的影响及临床意义.方法62例小儿先心病患者(男39例,女23例,体重9.5～20kg,年龄平均2.5岁)随机分成AB两组,A组用5%葡萄糖预充,B组无糖预充,各组再分成超滤组和对照组,两组病种基本相同.超滤组和对照组在复温前、停机时,分别采血和超滤液测定血糖和乳酸,进行对比.结果A组和B组超滤后血清中血糖相比(P＜0.01),两组统计学差异有显著性.超滤组与对照组血糖和乳酸浓度,无明显统计学差异.而超滤液中含有较高浓度糖和乳酸.结论无糖预充可以降低血糖和乳酸,用超滤法可直接排除心内直视术小儿患者高血糖和高乳酸,减少神经系统的受损,值得临床推广应用.     

Effect of parathyroidin on the course of acute myocardial ischemia in experiments on rats

Experiments were made on 187 white rats weighing 180-200 g. Acute myocardial ischemia was simulated in 137 animals. Fifty sham-operated rats served as control. Experimental acute myocardial ischemia was accompanied by an increase in blood creatine phosphokinase and lactate dehydrogenase activity as well as by an elevation of serum lactate level and fall of blood plasma calcium concentration, suppression of diuresis and excretion of calcium with urine. Intraperitoneal injections of parathyroidine to rats with acute myocardial ischemia led to rapid normalization of the homeostatic parameters. Lethality of experimental animals decreased 1.8-fold.     

Isoproterenol fails to produce myocardial necrosis in streptozotocin-induced diabetic rats.

Biochemical alterations in the hearts of non-diabetic and 5 weeks of streptozotocin-induced diabetic rats following isoproterenol (ISO) administration were compared. Serum lactate dehydrogenase (LDH) and myocardial adenosine triphosphate (ATP), creatine phosphate (CP), lactate and glycogen were used as indices of myocardial injury. Hearts from diabetic rats (blood glucose greater than 350 mg/dl), before ISO administration, had normal lactate levels but significantly low high-energy phosphate (HEP) levels and high glycogen levels in comparison to non-diabetic rats. No difference was observed in serum LDH levels between these two groups. ISO administration to non-diabetic rats caused myocardial necrosis as evidenced by a significant depletion of myocardial glycogen and HEPs along with significant myocardial lactate accumulation and an increase in serum LDH. However, the hearts from diabetic rats failed to show any significant HEP depletion, accumulation of lactate and leakage of LDH into serum following ISO-administration, though myocardial glycogen level was significantly lowered. These observations, in conjunction with earlier reports, point to the hypothesis that, in diabetes, there are certain alterations in the sarcolemma which hamper the process by which ISO causes myocardial necrosis.     

Acute and remote biochemical and physiological effects of exhaustive weightlifting exercise

The goal of the work was a study of exhaustive weightlifting exercise effect on prolonged changes in physiological and biochemical variables characterized functional status of skeletal muscles. An exercise gave rise to significant blood lactate concentration increase that was indicative of an anaerobic metabolism to be a predominant mechanism of muscle contraction energy supply. A reduction of m. rectus femoris EMG activity (amplitude and frequency), tonus of tension and an increase in tonus of relaxation were found immediately after exercise. Both EMG amplitude and frequency were increased 1 day post-exercise. However, after 3 days of recovery, EMG amplitude and frequency were decreased again and, in parallel, blood serum creatine kinase (CK) activity was significantly increased. After 9 recovery days, all measured variables with the exception of CK were normalized. A significant reverse correlation was found between blood serum lactate concentration and m. rectus femoris EMG activity at the same time points. Blood serum CK activity and m. rectus femoris EMG and tonus variables were observed to be significantly reversely correlated on the 3rd post-exercise day. Presented data demonstrate that exhaustive exercise-induced muscle injury resulted in phase alterations in electrical activity and tonus which correlated with lactate concentration and CK activity in blood serum.     

Lacticacidosis: A Clinically Significant Aspect of Shock

A study was made of the metabolic acidosis of hypotensive shock in 25 patients in an attempt to elucidate its etiology and to determine if the degree of acidosis might be a good parameter for the evaluation of treatment and prognosis.Blood lactate was elevated (> 1.3 mEq./l.) in 24 of 25 patients in hypotensive shock. There was a good correlation (r= 0.83, p < 0.01) between rising blood lactate and decrease in serum bicarbonate and arterial pH, early in shock. These data indicate that the metabolic acidosis of early shock is largely due to lactate ion. Evidence is presented that high blood lactate levels early in shock are indicative of poor prognosis.     

A 44-kDa of protein identical to the N-terminal amino acid sequence of MCT1 in human circulation

A family of specific carrier protein designated as monocarboxylate transporter (MCT) has been known to transport the lactate and other moncarboxylates in mammalian cells. We hypothesized the presence of serum protein in human circulation that may works as a lactate carrier and that biochemical structure would possesses common structure with MCT on the plasma membrane.Immunoblot analysis with an anti-MCT1 polyclonal antibody suggested the presence of a 44-kDa protein in human circulation and N-terminal amino acid sequencing exhibited a stretch of 14 amino acids which is completely identical to MCT1. The unbound fractions from the GST-MCT1 fusion protein-immobilized glutathione sepharose 4B column demonstrated that lactic acid concentration began to increase with one fraction delay compared to Sepharose 4B and GST-immobilized column. When lactic acid was washed away with PBS, lactic acid concentrations in the effuluent constantly decreased in both Sepharose 4B and GST-immobilized column. However, GST-MCT1-immobilized column showed specific convex curve from fraction approximately 3 mM of lactate and demonstrated wash out delay compared to Sepharose 4B and GST-immobilized column.These observations demonstrated biochemical and immunological similarities between a 44-kDa protein purified from human serum and MCT1 present on the plasma membrane. The studies on MCT1-fusion protein suggested possible functional properties of a 44-kDa protein as a lactate buffer by holding and unhand a lactate according to the lactate concentration in human blood. The experiments described herein have suggested the existence of lactate carrier in human circulation which is free from plasma membrane.     

Activation of carboxylation as a factor in correcting the metabolic disorder in diabetes mellitus

The paper deals with studies of the effect of the 10-day complex therapy with carbostimulin application on the content of the tricarboxylic cycle and glycolysis metabolites, total lipids, cholesterol, triglycerides in blood and potassium and sodium in blood serum of patients with diabetes mellitus. It is established that under the effect of carbostimulin the content of oxaloacetate, lactate and alpha-glycerophosphate becomes normal, the lactate/pyruvate ratio elevated in diabetes decreases, which evidences for intensification of reduction processes in the cytoplasm.     

Effects of a 30-km race upon salivary lactate correlation with blood lactate

Blood lactate has been used to determine the aerobic capacity and long distance performance. Recently, a new methodology has been suggested to supplant the invasive blood lactate techniques. Salivary lactate has received attention because it shows high correlation to blood lactate in progressive overload test. We evaluated the correlation between salivary and blood lactate during a long distance run and assessed possible changes in salivary lactate concentration. Fifteen expert marathon racers ran 30 km as fast as possible. Saliva and 25 muL of blood were collected at rest and at each 6 km for lactate determination. Blood lactate concentration increased in the 6th km and then remained constant until the end of the race. Salivary lactate increased after 18 km in relation to basal. We found high correlations between blood and saliva absolute lactate (r=0.772, p<0.05) and the blood lactate relative concentration corrected by protein (r=0.718, p<0.05). The highest correlation found between absolute and relative salivary lactate was r=0.994 (p<0.001). Our results show that it is possible to use salivary lactate with absolute values or relative protein concentration. In addition, salivary lactate showed a high correlation with blood lactate in endurance events.     

Ease of calving, blood chemistry, insulin and bovine growth hormone of newborn calves derived from embryos produced in vitro in culture systems with serum and co-culture or with PVA

Blood chemistry (pH, pCO2, pO2, glucose, lactate) as well as plasma insulin and growth hormone of calves derived from embryos produced under 2 different in vitro culture systems (modified SOFaa with 20% serum and co-culture with bovine oviduct epithelial cells [IVP serum, n=8] or with 3 mg/mL PVA [IVPdefined, n=6]) were compared with those of calves derived from AI (n=5). Calvings were classified according to the ease (unassisted, light traction, heavy traction). Blood samples were taken from the jugular vein of calves at 5, 15, 30 and 60 min, and at 2, 3, 6, 12, 18 and 24 h after delivery, then daily for 6 d. At the second day of life after 4 feedings and a 4-h fasting period, a glucose tolerance test was performed to evaluate glucose metabolism and insulin secretion. Calves in the IVP serum group had higher birth weights than AI calves (LS mean +/- SEM, IVP serum: 45.2 +/- 1.4 kg vs AI: 40.4 +/- 1.7 kg; P < 0.05), while the birth weights of calves in the IVP defined group were in between (IVPdefined: 41.9 +/- 1.6 kg). More IVP serum calves (75%) needed assistance than IVP defined (33%) or AI (40%) calves. The effect of ease of calving vs the effect of embryo culture was compared in relation to blood parameters at birth. There was an effect of ease of calving but not of embryo culture conditions on blood pH, lactate and PCO2. Calves requiring heavy traction had lower pH during the first 3 h after calving, a higher lactate during the first 60 min after calving and a higher pCO2 the first 2 h after calving than calves born unassisted. Calves requiring heavy traction also had lower pH the first 2 h and higher lactate the first 3 h after calving than calves born by light traction. IVP defined calves had lower lactate than IVP serum calves the first 60 min after calving. At 6 h after delivery, all blood parameters had stabilized. There was no effect of either embryo culture or ease of calving on basal insulin and growth hormone level, or the ability of the calves to handle glucose postnatally and during a glucose tolerance test.     

Blood lactate production and recovery from anaerobic exercise in trained and untrained boys

Blood lactate production and recovery from anaerobic exercise were investigated in 19 trained (AG) and 6 untrained (CG) prepubescent boys. The exercises comprised 3 maximal test performances; 2 bicycle ergometer tests of different durations (15 s and 60 s), and running on a treadmill for 23.20 +/- 2.61 min to measure maximal oxygen uptake. Blood samples were taken from the fingertip to determine lactate concentrations and from the antecubital vein to determine serum testosterone. Muscle biopsies were obtained from vastus lateralis. Recovery was passive (seated) following the 60 s test but that following the treadmill run was initially active (10 min), and then passive. Peak blood lactate was highest following the 60 s test (AG, 13.1 +/- 2.6 mmol.1-1 and CG, 12.8 +/- 2.3 mmol.1-1). Following the 15 s test and the treadmill run, peak lactate values were 68.7 and 60.6% of the 60 s value respectively. Blood lactate production was greater (p less than 0.001) during the 15 s test (0.470 +/- 0.128 mmol.1-1.s-1) than during the 60 s test (0.184 +/- 0.042 mmol.1-1.s-1). Although blood lactate production was only nonsignificantly greater in AG, the amount of anaerobic work in the short tests was markedly greater (p less than 0.05-0.01) in AG than CG. Muscle fibre area (type II%) and serum testosterone were positively correlated (p less than 0.05) with blood lactate production in both short tests. Blood lactate elimination was greater (p less than 0.001) at the end of the active recovery phase than in the next (passive) phase.(ABSTRACT TRUNCATED AT 250 WORDS)     

Lactate dehydrogenase and its isoenzymes in human umbilical cord tissue]

Study on umbilical cord tissue, completely bloodless, for determination of lactate deshydrogenase activity and its distribution among the five iso-enzymes. Comparison with placenta, amniotic fluid, serums of blood of cord and of mother. Cord tissue is very active (about 360 muKatals, in average) and it is a similar result in placenta (as it is possibly bloodless). Blood serum of cord is more active than amniotic fluid, which is more active than maternal serum, but they are 80 to 200 times less active than cord tissue. After electrophoresis, a very large predominance of the slow iso-enzymes L.D.H. 4--5 is found in cord tissue (72%), amniotic fluid (67%) and placenta (56%), whereas the fast iso-enzymes L.D.H. 1--2 are predominant in the serums of cord blood and of mother. These data indicate an intense metabolic activity in the cord tissue, which has also an high level of lactate, and this seems related to the foetal metabolism for anaerobic glycolysis in oxygen weakly provided tissues.     

Lactate Dehydrogenase Activity and Insulin and Lactate Levels at an Altitude Below Sea Level (–350 m) Compared to Those at an Altitude Above Sea Level (620 m) After Exercise

This study was designed to investigate the effect of exercise at 350 m below sea level altitude (–350 m) on the serum levels of lactate dehydrogenase (LDH), insulin, and lactate. The study was carried out on ten trained adult males with mean age of 23.3 ± 3.4 years following a 21-km noncompetitive run. Venous blood was withdrawn from the subjects before exercise and 5 min post exercise. For comparison purposes, a similar study was performed with the same subjects but at 620 m above sea level (+620 m). The results show a significant increase in LDH and lactate levels after exercise only at low altitude (–350 m). Serum insulin levels decreased significantly after exercise at both altitudes. These changes in serum levels of LDH, insulin, and lactate at different altitudes suggest that a type of metabolic adjustment is present that meets energy requirements during exercise.     

The effect of different blood sampling sites and analyses on the relationship~ between exercise intensity and 4.0 mmol ·1? blood lactate concentration

The aim of the study was to examine whether the difference in lactate concentration in different blood fractions is of practical importance when using blood lactate as a test variable of aerobic endurance capacity. Ten male firefighters performed submaximally graded exercise on a cycle ergometer for 20-25 min. Venous and capillary blood samples were taken every 5 min for determination of haematocrit and lactate concentrations in plasma, venous and capillary blood. At the same time, expired air was collected in Douglas bags for determination of the oxygen consumption. A lactate concentration of 4.0 mmol.l-1 was used as the reference value to compare the oxygen consumption and exercise intensity when different types of blood specimen and sampling sites were used for lactate analysis. At this concentration the exercise intensity was 17% lower (P less than 0.01) when plasma lactate was compared to venous blood lactate, and 12% lower (P less than 0.05) when capillary blood lactate was used. Similar discrepancies were seen in oxygen consumption. The results illustrated the importance of standardizing sampling and handling of blood specimens for lactate determination to enable direct comparisons to be made among results obtained in different studies.     

Investigation of dye/protein interaction and its application to enzyme purification

In this review the results of the interaction of the active dyes used in the USSR textile industry with microbial enzymes and blood serum proteins are discussed. The complexity of dye/protein interaction and the dependence of this interaction on different factors is demonstrated. Some practical aspects of the use of dye containing sorbents are presented and discussed. Their suitability for RNA ligase and DNA ligase, acetate kinase, alcohol dehydrogenase, lactate dehydrogenase and glucose-6-phosphate dehydrogenase purification and blood serum protein fractionation is demonstrated.     

Insulin and non-esterified fatty acids. Acute regulators of lipogenesis in perfused rat liver.

Livers from fed rats were perfused with whole rat blood and infused with oleate to maintain constant concentrations of serum non-esterified fatty acids over a wide physiological range. Infusion of insulin opposed the antilipogenic effects of increasing concentrations of serum non-esterified fatty acids. Secretion of very-low-density-lipoprotein triacylglycerols was directly proportional to the concentration of serum non-esterified fatty acids and was increased by insulin. The secretion of newly-synthesized fatty acids in very-low-density-lipoprotein triacylglycerols decreased with increasing concentrations of serum non-esterified fatty acid. Insulin opposed this change. Cholesterol biosynthesis was unaffected by alterations in concentration of serum non-esterified fatty acid but was increased by insulin. Equilibrium concentrations of perfusate lactate and glucose were increased by serum non-esterified fatty acids but steady-state rates of hepatic glucose output and lactate uptake were unchanged. Insulin decreased perfusate glucose concentrations and abolished the increase in its concentration that resulted from increases in non-esterified fatty acid concentrations.     

The blood chemical status of Atlantic cod Gadus morhua following capture by jig and demersal longline with differential hook removal methods

Common haematological [haematocrit (Hct)], primary (serum cortisol) and secondary (serum glucose and plasma lactate) analytes were utilized to compare blood biochemical status of Gadus morhua captured rapidly by jig with that of G. morhua captured by commercial demersal longline. In general, the physiological status of G. morhua, despite blind hook times, was significantly more disrupted (pronounced haemo-concentration and significantly elevated concentrations of cortisol, glucose and lactate) following longline capture relative to capture by jig, while no differences were detected among longline-caught fish as a function of dehooking method (or concomitant extent of overt physical trauma). Blood profiles from the more stressed G. morhua, a possible function of more extended longline hook times, were similar to the most stressed values reported for this species. The results also demonstrate that, although acute blood biochemical status is an effective gauge of relative stress, it does not reflect physical injury status, which has been shown to exert a strong influence on delayed mortality in previous studies in this species. Thus, acute blood chemical status alone may not be the most complete predictor of mortality. Future studies should evaluate physiological repercussions from capture-handling against physical trauma during more extended post-release periods for this species.     

Novel system for real-time ex vivo lactate monitoring in human whole blood

The objective of the study was to evaluate the performance of an amperometric enzyme based lactate sensor and to investigate the possibility of replacing a double lumen catheter based blood withdrawal system with a heparin coated single lumen system. The inner lumen of a double lumen catheter which was placed in a peripheral vein was perfused with heparin solution. The outer lumen was used to collect heparinized blood samples at a defined flow rate. The single lumen system was attached to a heparinized catheter which was also placed in a peripheral vein. The undiluted blood samples were collected at a specified flow rate. A sensor flow chamber incorporating an amperometric thin-film lactate microbiosensor was placed in the sampling line for real-time lactate monitoring. Plasma lactate concentrations were measured during frequently performed hyperlactatemia bicycle ergometer experiments in six healthy volunteers (age 25.8±2.8 years, BMI 22.7±1 kg/m²). Additionally, plasma lactate was measured in real-time using the lactate sensors. The first three experiments were performed with a double lumen based catheter system whereas the following three experiments were performed with a heparin coated catheter system. The correlation coefficients of sensor readings and laboratory analyzer results in all six experiments were between 0.93 and 0.99, respectively (P<0.001). The miniaturized lactate sensors showed a linear range up to 25 mmol/l lactate concentration and 95% response times <30 s in undiluted serum. During the experiments maximum lactate concentrations of 14 mmol/l were achieved. Improvements of system performance using heparin coated catheter systems could be shown. The overall SD of the sensor readings compared to laboratory results using three double lumen catheter based systems was 0.91 mmol/l whereas the SD using three heparin coated systems was 0.65 mmol/l. In summary, real-time monitoring of lactate in human whole blood is feasible with such a device and can be improved by using heparin coated catheter systems.     

Oxamate Improves Glycemic Control and Insulin Sensitivity via Inhibition of Tissue Lactate Production in db/db Mice

Oxamate (OXA) is a pyruvate analogue that directly inhibits the lactate dehydrogenase (LDH)-catalyzed conversion process of pyruvate into lactate. Earlier and recent studies have shown elevated blood lactate levels among insulin-resistant and type 2 diabetes subjects and that blood lactate levels independently predicted the development of incident diabetes. To explore the potential of OXA in the treatment of diabetes, db/db mice were treated with OXA in vivo. Treatment of OXA (350–750 mg/kg of body weight) for 12 weeks was shown to decrease body weight gain and blood glucose and HbA1c levels and improve insulin secretion, the morphology of pancreatic islets, and insulin sensitivity in db/db mice. Meanwhile, OXA reduced the lactate production of adipose tissue and skeletal muscle and serum lactate levels and decreased serum levels of TG, FFA, CRP, IL-6, and TNF-α in db/db mice. The PCR array showed that OXA downregulated the expression of Tnf, Il6, leptin, Cxcr3, Map2k1, and Ikbkb, and upregulated the expression of Irs2, Nfkbia, and Pde3b in the skeletal muscle of db/db mice. Interestingly, LDH-A expression increased in the islet cells of db/db mice, and both treatment of OXA and pioglitazone decreased LDH-A expression, which might be related to the improvement of insulin secretion. Taken together, increased lactate production of adipose tissue and skeletal muscle may be at least partially responsible for insulin resistance and diabetes in db/db mice. OXA improved glycemic control and insulin sensitivity in db/db mice primarily via inhibition of tissue lactate production. Oxamic acid derivatives may be a potential drug for the treatment of type 2 diabetes.