Fructose-induced ROS generation impairs glucose utilization in L6 skeletal muscle cells

Abstract

High fructose consumption has implicated in insulin resistance and metabolic syndrome. Fructose is a highly lipogenic sugar that has intense metabolic effects in liver. Recent evidences suggest that fructose exposure to other tissues has substantial and profound metabolic consequences predisposing toward chronic conditions such as type 2 diabetes. Since skeletal muscle is the major site for glucose utilization, in the present study we define the effects of fructose exposure on glucose utilization in skeletal muscle cells. Upon fructose exposure, the L6 skeletal muscle cells displayed diminished glucose uptake, glucose transporter type 4 (GLUT4) translocation, and impaired insulin signaling. The exposure to fructose elevated reactive oxygen species (ROS) production in L6 myotubes, accompanied by activation of the stress/inflammation markers c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase 1/2 (ERK1/2), and degradation of inhibitor of NF-κB (IκBα). We found that fructose caused impairment of glucose utilization and insulin signaling through ROS-mediated activation of JNK and ERK1/2 pathways, which was prevented in the presence of antioxidants. In conclusion, our data demonstrate that exposure to fructose induces cell-autonomous oxidative response through ROS production leading to impaired insulin signaling and attenuated glucose utilization in skeletal muscle cells.     

Postnatal exposure to di-(2-ethylhexyl)phthalate alters cardiac insulin signaling molecules and GLUT4Ser488 phosphorylation in male rat offspring

Di-(2-ethylhexyl)phthalate (DEHP), a distinctive endocrine-disrupting chemical, is widely used as a plasticizer in a variety of consumer products. It can easily cross the placenta and enter breast milk and then it is rapidly absorbed by offspring. Since it is generally accepted that individuals are more sensitive to chemical exposure during vital developmental periods, we investigated whether DEHP exposure during lactation affects cardiac insulin signaling and glucose homeostasis in the F₁ male rat offspring at postnatal day 22 (PND22). Lactating Wistar rats were administered with DEHP (1, 10, and 100 mg/kg/d) or olive oil from lactation day 1 to 21 by oral gavage. All the male pups were perfused and killed on PND22. On the day before the killing, they were kept for fasting overnight and blood was collected. The cardiac muscle was dissected out, washed in ice-cold physiological saline repeatedly and used for the assay of various parameters. DEHP-exposed offspring had significantly lower body weight than the control. DEHP-exposed offspring showed elevated blood glucose, decreased ¹⁴C-2-deoxyglucose uptake and ¹⁴C-glucose oxidation in cardiac muscle at PND22. The concentration of upstream insulin signaling molecules such as insulin receptor subunit β (InsRβ) and insulin receptor substrate 1 (IRS1) were downregulated in DEHP-exposed offspring. However, no significant alterations were observed in protein kinase B (Akt) and Akt substrate of 160 kDa (AS160). Surprisingly, phosphorylation of IRS1 ^Tyr632 and Akt ^Ser473 were diminished. Low levels of glucose transporter type 4 (GLUT4) protein and increased GLUT4 ^Ser488 phosphorylation which decreases its intrinsic activity and translocation towards plasma membrane were also recorded. Lactational DEHP exposure predisposes F ₁ male offspring to cardiac glucometabolic disorders at PND22, which may impair cardiac function.     

Comparison of Effects of Sublethal Microwave Radiation and Conventional Heating on the Metabolic Activity of Staphylococcus aureus

This study was conducted in an attempt to characterize some of the effects of sublethal microwave radiation on cells of Staphylococcus aureus. Cultures were exposed to microwave radiation for 10, 20, 30, and 40 s. The effects of a conventional heat treatment were also compared by placing flasks containing cultures in a boiling water bath for the amount of time required to reach temperatures equivalent to those found in cultures exposed to microwave radiation. Control, microwave-treated, and conventionally heat-treated cultures were centrifuged, pellets were resuspended in distilled water, and the resulting suspensions were passed through a French pressure cell. Cell lysates and walls were then isolated and assayed for enzymatic activity. Thermonuclease production was also determined at various levels of exposure of cells to microwave radiation. Activities of malate and α-ketoglutarate dehydrogenases, cytochrome oxidase, and cytoplasmic adenosine triphosphatase were higher in microwave-treated cells than in control cells. Membrane adenosine triphosphatase, alkaline phosphatase, and lactate dehydrogenase activities were unaffected when cells were exposed to microwave radiation. The activity of glucose-6-phosphate dehydrogenase was decreased by exposure of cells to microwave radiation. In conventionally heated cells, activities of glucose-6-phosphate and malate dehydrogenases and cytoplasmic adenosine triphosphatase increased activities of α-ketoglutarate and lactate dehydrogenases decreased, and alkaline phosphatase activity remained unaffected. Increased levels of thermonuclease activity were observed when cells were exposed to microwave radiation for 10 or 20 s. Data indicate that microwave radiation affects S. aureus in a manner which cannot be explained solely by thermal effects.     

Evidence That Calcium Release-activated Current Mediates the Biphasic Electrical Activity of Mouse Pancreatic β-Cells

The electrical response of pancreatic β-cells to step increases in glucose concentration is biphasic, consisting of a prolonged depolarization with action potentials (Phase 1) followed by membrane potential oscillations known as bursts. We have proposed that the Phase 1 response results from the combined depolarizing influences of potassium channel closure and an inward, nonselective cation current (I _CRAN) that activates as intracellular calcium stores empty during exposure to basal glucose (Bertram et al., 1995). The stores refill during Phase 1, deactivating I _CRAN and allowing steady-state bursting to commence. We support this hypothesis with additional simulations and experimental results indicating that Phase 1 duration is sensitive to the filling state of intracellular calcium stores. First, the duration of the Phase 1 transient increases with duration of prior exposure to basal (2.8 mm) glucose, reflecting the increased time required to fill calcium stores that have been emptying for longer periods. Second, Phase 1 duration is reduced when islets are exposed to elevated K⁺ to refill calcium stores in the presence of basal glucose. Third, when extracellular calcium is removed during the basal glucose exposure to reduce calcium influx into the stores, Phase 1 duration increases. Finally, no Phase 1 is observed following hyperpolarization of the β-cell membrane with diazoxide in the continued presence of 11 mm glucose, a condition in which intracellular calcium stores remain full. Application of carbachol to empty calcium stores during basal glucose exposure did not increase Phase 1 duration as the model predicts. Despite this discrepancy, the good agreement between most of the experimental results and the model predictions provides evidence that a calcium release-activated current mediates the Phase 1 electrical response of the pancreatic β-cell. Received: 5 June 1996/Revised: 15 August 1996     

The Effect of Microwave Radiation on Binding of Propyl-2,3-[3H]-Dehydroalprenololto β-Adrenergic Receptors of Rat Erythrocyte Membranes

The effect of microwave radiation (2450 MHz) on binding of the ligand propyl-2,3-[³H]-dehydroalprenolol to β-adrenergic receptors of rat erythrocyte membranes has been studied. It is shown that microwave exposure decreases specific binding of the ligand to receptors. The effect is dependent on time of exposure (66.2% on 10 min exposure and 37.3% on 30 min exposure). EDTA and heparin used as anticoagulants do not significantly alter the experimental results. However, the Ca²⁺ content in the incubation medium is essential for the effect. Exposure of erythrocytes isolated in the presence of EDTA and incubated in Hanks' solution without CA²⁺ has no effect on binding. The effect is manifest when Ca²⁺ is added to the specimen exposed. Possible mechanisms of action of microwaves on the studied parameter are discussed.     

Immunologic and hematopoietic alterations by 2,450-MHz electromagnetic radiation

A biphasic modulation of responsiveness of spleen lymphocytes to mitogens was observed in mice exposed to 2,450-MHz radiation at power densities of 5–15 mW/cm² over various periods ranging between one and 17 days. This modulated phenomenon may be explained on the basis of 1) suppression of lymphocyte response by microwave-activated macrophages which persists throughout the entire course of radiation, and 2) concurrent progressive direct stimulation of lymphocytes which culminates around day 9 of exposure. Tumor cytotoxicity of killer lymphocytes from mice exposed to five or nine days of radiation did not appear different from sham controls. The highly proliferative hematopoietic marrow cells were sensitive to microwave radiation. Nine days of exposure to radiation (15 mW/cm²) reduced the colonyforming units of myeloid and erythroid series by 50%. This observation may offer a new and more sensitive assay for studying biological effects of electromagnetic radiation.     

Micronucleus induction after whole-body microwave irradiation of rats

Adult male Wistar rats were exposed for 2 h a day, 7 days a week for up to 30 days to continuous 2450 MHz radiofrequency microwave (rf/MW) radiation at a power density of 5–10 mW/cm². Sham-exposed rats were used as controls. After ether anesthesia, experimental animals were euthanized on the final irradiation day for each treated group. Peripheral blood smears were examined for the extent of genotoxicity, as indicated by the presence of micronuclei in polychromatic erythrocytes (PCEs). The results for the time-course of PCEs indicated significant differences (P<0.05) for the 2nd, the 8th and the 15th day between control and treated subgroups of animals. Increased influx of immature erythrocytes into the peripheral circulation at the beginning of the experiment revealed that the proliferation and maturation of nucleated erythropoietic cells were affected by exposure to the 2450 MHz radiofrequency radiation. Such findings are indicators of radiation effects on bone-marrow erythropoiesis and their subsequent effects in circulating red cells. The incidence of micronuclei/1000 PCEs in peripheral blood was significantly increased (P<0.05) in the subgroup exposed to rf/MW radiation after eight irradiation treatments of 2 h each in comparison with the sham-exposed control group. It is likely that an adaptive mechanism, both in erythrocytopoiesis and genotoxicity appeared in the rat experimental model during the subchronic irradiation treatment.     

Significance of microthermal effects derived from low level UHF-microwave irradiation of the head: indirect caloric vestibular stimulation

Reports of behavioral and clinical changes following weak microwave irradiation, though not fully documented, are of sufficient moment to require examination of each possible biological consequence of low level exposure, particularly with respect to the central nervous system. In this report the hypothesis that significant cytological microthermal effects are induced by low intensity microwave fields (10 mW/cm² incident power density) is examined. An estimate of the upper bound on the thermal effects thus produced is made, showing the thermal variations to be no larger than those endogenous to neural tissue. A similar analysis of microthermal effects within the vestibulo-cochlear apparatus, however, suggests the more limited hypothesis that this structure is responsive to weak, absorbed microwave energy. An estimate of the temperature gradients, hence, local fluid density changes within the labyrinth supports the existence of detectable intralabyrinthine convective forces at incident power densities as low as 15–20 mW/cm². This suggests (i) that microwave induced vestibular effects may provide a cue to alert personnel to significant acute microwave exposure, (ii) that reports of behavioral and/or clinical reactivity to low level microwave exposure may derive from such a benign but potentially useful interaction and (iii) that geometric peculiarities of the vestibulo-cochlear apparatus may result in markedly enhanced microwave-labyrinthine coupling at particular radiation wavelengths.     

Effect of chronic microwave radiation on T cell-mediated immunity in the rabbit

Experiments were conducted to elucidate the effects of chronic low power-level microwave radiation on the immunological systems of rabbits. Fourteen male Belgian white rabbits were exposed to microwave radiation at 5 mW/cm², 2.1 GHz, 3 h daily, 6 days/week for 3 months in two batches of 7 each in specially designed miniature anechoicchambers. Seven rabbits were subjected to sham exposure for identical duration. The microwave energy was provided through S band standard gain horns connected to a 4K3SJ2 Klystron power amplifier. The first batch of animals were assessed for T lymphocyte-mediated cellular immune response mechanisms and the second batch of animals for B lymphocyte-mediated humoral immune response mechanisms. The peripheral blood samples collected monthly during microwave/sham exposure and during follow-up (5/14 days after termination of exposures, in the second batch animals only) were analysed for T lymphocyte numbers and their mitogen responsiveness to ConA and PHA. Significant suppression of T lymphocyte numbers was noted in the microwave group at 2 months (P<0.01, % 21.5%) and during follow-up (P<0.01, % 30.2%). The first batch animals were initially sensitised with BCG and challenged with tuberculin (0.03 ml) at the termination of microwave irradiation/sham exposure and the increase in foot pad thickness ( mm), which is a measure of T cell-mediated immunity (delayed type hypersensitivity response, DTH) was noted in both the groups. The microwave group revealed a better response than the control group (%+12.4 vs.+7.54). The animals were sacrified and the tissue T lymphocyte counts (spleen and lymph node) were analysed. No significant variation was observed in the tissue T lymphocyte counts of microwave-irradiated rabbits. From these results it is speculated that the T lymphocytes are sequestered to various lymphoid organs under the influence of microwaves. A sub-population of T cells known as T helper cells (mediating DTH response) are probably not affected by microwave radiation. It is clear from our experiments that although chronic microwave radiation at 5 mW/cm² leads to suppression of peripheral T lymphocyte numbers, there is no concomitant functional impairment of these cells as evidenced by functional assays.     

The Quantitative Relationship Between “The Metabolic Syndrome” and Abdominal Obesity in Women

Abdominal obesity is closely associated with the presence of metabolic risk factors and elevated blood pressure in selected materials. This has, however, never been analyzed quantitatively in a non-selected cohort. Therefore, in a population-based study of 1462 Swedish women, four selected risk factors for non-insulin dependent diabetes mellitus (NIDDM) and cardiovascular disease (CVD), serum triglycerides, blood glucose and systolic blood pressure and also serum insulin in a subsample, were examined in relation to regional and overall obesity. This was performed by subdividing the age adjusted sample into quintiles of waist to hip circumference ratio (WHR) or body mass index (BMI) as indicators of abdominal distribution of body fat and overall obesity, respectively. The risk factors serum triglycerides, blood glucose, blood pressure and serum insulin were defined as being elevated when the value of the risk factor was higher than the mean plus one or two standard deviations of the total age-adjusted cohort. The percentage of women with elevated risk factors according to this definition was then calculated in each of these quintiles. Having a risk factor which was elevated according to the definition was significantly correlated to WHR and BMI (p<0.0001) independent of age. The presence of one or several of these elevated risk factors was clearly higher than expected in the fifth quintile of WHR, and to a lesser extent in the fifth quintile of BMI while this was not the case in the lower quintiles of WHR and BMI. When studying the combination of the WHR and BMI, the presence of risk factors higher than the mean plus two standard deviations increased gradually with WHR in all five quintiles of BMI. A significant association was observed between WHR and presence of risk factors independent of BMI (p<0.0001) but BMI did not remain significantly correlated to presence of risk factors when controlling for WHR (p=0.09). These results indicate that abdominal distribution of body fat in women independently of general obesity is closely associated with metabolic risk factors including elevated blood pressure, a metabolic syndrome with increased risk for cardiovascular disease and non-insulin dependent diabetes mellitus.     

Reduction of Phosphorylated Synapsin I (Ser-553) Leads to Spatial Memory Impairment by Attenuating GABA Release after Microwave Exposure in Wistar Rats

Background

Abnormal release of neurotransmitters after microwave exposure can cause learning and memory deficits. This study investigated the mechanism of this effect by exploring the potential role of phosphorylated synapsin I (p-Syn I).

Methods

Wistar rats, rat hippocampal synaptosomes, and differentiated (neuronal) PC12 cells were exposed to microwave radiation for 5 min at a mean power density of 30 mW/cm². Sham group rats, synaptosomes, and cells were otherwise identically treated and acted as controls for all of the following post-exposure analyses. Spatial learning and memory in rats was assessed using the Morris Water Maze (MWM) navigation task. The protein expression and presynaptic distribution of p-Syn I and neurotransmitter transporters were examined via western blotting and immunoelectron microscopy, respectively. Levels amino acid neurotransmitter release from rat hippocampal synaptosomes and PC12 cells were measured using high performance liquid chromatograph (HPLC) at 6 hours after exposure, with or without synapsin I silencing via shRNA transfection.

Results

In the rat experiments, there was a decrease in spatial memory performance after microwave exposure. The expression of p-Syn I (ser-553) was decreased at 3 days post-exposure and elevated at later time points. Vesicular GABA transporter (VGAT) was significantly elevated after exposure. The GABA release from synaptosomes was attenuated and p-Syn I (ser-553) and VGAT were both enriched in small clear synaptic vesicles, which abnormally assembled in the presynaptic terminal after exposure. In the PC12 cell experiments, the expression of p-Syn I (ser-553) and GABA release were both attenuated at 6 hours after exposure. Both microwave exposure and p-Syn I silencing reduced GABA release and maximal reduction was found for the combination of the two, indicating a synergetic effect.

Conclusion

p-Syn I (ser-553) was found to play a key role in the impaired GABA release and cognitive dysfunction that was induced by microwave exposure.     

Insulin secretion kinetics from single islets reveals distinct subpopulations

Type II diabetes progresses with inadequate insulin secretion and prolonged elevated circulating glucose levels. Also, pancreatic islets isolated for transplantation or tissue engineering can be exposed to glucose over extended timeframe. We hypothesized that isolated pancreatic islets can secrete insulin over a prolonged period of time when incubated in glucose solution and that not all islets release insulin in unison. Insulin secretion kinetics was examined and modeled from single mouse islets in response to chronic glucose exposure (2.8‐20 mM). Results with single islets were compared to those from pools of islets. Kinetic analysis of 58 single islets over 72 h in response to elevated glucose revealed distinct insulin secretion profiles: slow‐, fast‐, and constant‐rate secretors, with slow‐secretors being most prominent (ca., 50%). Variations in the temporal response to glucose therefore exist. During short‐term (<4 h) exposure to elevated glucose few islets are responding with sustained insulin release. The model allowed studying the influence of islet size, revealing no clear effect. At high‐glucose concentrations, when secretion is normalized to islet volume, the tendency is that smaller islets secrete more insulin. At high‐glucose concentrations, insulin secretion from single islets is representative of islet populations, while under low‐glucose conditions pooled islets did not behave as single ones. The characterization of insulin secretion over prolonged periods complements studies on insulin secretion performed over short timeframe. Further investigation of these differences in secretion profiles may resolve open‐ended questions on pre‐diabetic conditions and transplanted islets performance. This study deliberates the importance of size of islets in insulin secretion. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1059–1068, 2018     

Insulin‐ and Exercise‐Stimulated Skeletal Muscle Blood Flow and Glucose Uptake in Obese Men

Objective: Insulin resistance in obese subjects results in the impaired use of glucose by insulin‐sensitive tissues, e.g., skeletal muscle. In the present study, we determined whether insulin resistance in obesity is associated with an impaired ability of exercise to stimulate muscle blood flow, oxygen delivery, or glucose uptake. Research Methods and Procedures: Nine obese (body mass index = 36 ± 2 kg/m²) and 11 age‐matched nonobese men (body mass index = 22 ± 1 kg/m²) performed one‐legged isometric exercise during hyperinsulinemia. Rates of femoral muscle blood flow, oxygen consumption, and glucose uptake were measured simultaneously in both legs using [¹⁵O]H₂O, [¹⁵O]O₂, [¹⁸F]fluoro‐deoxy‐glucose, and positron emission tomography. Results: The obese subjects exhibited resistance to insulin stimulation of glucose uptake in resting muscle, regardless of whether glucose uptake was expressed per kilogram of femoral muscle mass (p = 0.001) or per the total mass of quadriceps femoris muscle. At similar workloads, oxygen consumption, blood flow, and glucose uptake were lower in the obese than the nonobese subjects when expressed per kilogram of muscle, but similar when expressed per quadriceps femoris muscle mass. Discussion: We conclude that obesity is characterized by insulin resistance of glucose uptake in resting skeletal muscle regardless of how glucose uptake is expressed. When compared with nonobese individuals at similar absolute workloads and under identical hyperinsulinemic conditions, the ability of exercise to increase muscle oxygen uptake, blood flow, and glucose uptake per muscle mass is blunted in obese insulin‐resistant subjects. However, these defects are compensated for by an increase in muscle mass.     

Microwave radiation (2.45 GHz)-induced oxidative stress: Whole-body exposure effect on histopathology of Wistar rats

Man-made microwave and radiofrequency (RF) radiation technologies have been steadily increasing with the growing demand of electronic appliances such as microwave oven and cell phones. These appliances affect biological systems by increasing free radicals, thus leading to oxidative damage. The aim of this study was to explore the effect of 2.45 GHz microwave radiation on histology and the level of lipid peroxide (LPO) in Wistar rats. Sixty-day-old male Wistar rats with 180 ± 10 g body weight were used for this study. Animals were divided into two groups: sham exposed (control) and microwave exposed. These animals were exposed for 2 h a day for 35 d to 2.45 GHz microwave radiation (power density, 0.2 mW/cm²). The whole-body specific absorption rate (SAR) was estimated to be 0.14 W/kg. After completion of the exposure period, rats were sacrificed, and brain, liver, kidney, testis and spleen were stored/preserved for determination of LPO and histological parameters. Significantly high level of LPO was observed in the liver (p < 0.001), brain (p < 0.004) and spleen (p < 0.006) in samples from rats exposed to microwave radiation. Also histological changes were observed in the brain, liver, testis, kidney and spleen after whole-body microwave exposure, compared to the control group.

Based on the results obtained in this study, we conclude that exposure to microwave radiation 2 h a day for 35 d can potentially cause histopathology and oxidative changes in Wistar rats. These results indicate possible implications of such exposure on human health.     

Fluorescence depolarization studies of red cell membrane fluidity. The effect of exposure to 1.0-GHz microwave radiation

The internal viscosity of human red blood cell membranes was investigated during exposure to continuous wave 1.0-GHz microwave radiation using fluorescence measurements of a lipid seeking molecular probe, diphenylhexatriene. Samples were exposed in a Crawford cell arranged so that fluorescence was measured during microwave exposure; specific absorption rates calculated from electrical measurements were approximately 0.6, 2 and 15 W/kg. Measurements were obtained at selected temperatures between 15 °C and 40 °C and as a function of the duration of exposure at 23 °C. Arrhenius-type plots of the temperature profile data were linear and showed no difference between exposed and control samples. The exposure duration data also showed no difference between exposed and control samples except for a small effect of elevated temperature at the highest exposure. The activation energy for motion of the fluorescent probe in its environment within the membrane lipid was not affected by the application of the microwave energy and no evidence for a lipid phase transition was found. These results indicate that the increased cation efflux from red cells, observed by others at certain transition temperatures during microwave exposure, was more likely to have been caused by alteration of the membrane bound protein than by changes in the lipid constituents of the red cell membrane.     

Drosophila oogenesis as a bio-marker responding to EMF sources

The model biological organisms Drosophila melanogaster and Drosophila virilis have been utilized to assess effects on apoptotic cell death of follicles during oogenesis and reproductive capacity (fecundity) decline. A total of 280 different experiments were performed using newly emerged flies exposed for short time daily for 3–7?d to various EMF sources including: GSM 900/1800?MHz mobile phone, 1880–1900?MHz DECT wireless base, DECT wireless handset, mobile phone-DECT handset combination, 2.44?GHz wireless network (Wi-Fi), 2.44?GHz blue tooth, 92.8?MHz FM generator, 27.15?MHz baby monitor, 900?MHz CW RF generator and microwave oven’s 2.44?GHz RF and magnetic field components. Mobile phone was used as a reference exposure system for evaluating factors considered very important in dosimetry extending our published work with D. melanogaster to the insect D. virilis. Distance from the emitting source, the exposure duration and the repeatability were examined. All EMF sources used created statistically significant effects regarding fecundity and cell death-apoptosis induction, even at very low intensity levels (0.3?V/m blue tooth radiation), well below ICNIRP’s guidelines, suggesting that Drosophila oogenesis system is suitable to be used as a biomarker for exploring potential EMF bioactivity. Also, there is no linear cumulative effect when increasing the duration of exposure or using one EMF source after the other (i.e. mobile phone and DECT handset) at the specific conditions used. The role of the average versus the peak E-field values as measured by spectrum analyzers on the final effects is discussed.     

Regulation of glucose utilization and lipogenesis in adipose tissue of diabetic and fat fed animals: Effects of insulin and manganese

In order to evaluate the modulatory effects of manganese, high fat diet fed and alloxan diabetic rats were taken and the changes in the glucose oxidation, glycerol release and effects of manganese on these parameters were measured from adipose tissue. An insulin-mimetic effect of manganese was observed in the adipose tissue in the controls and an additive effect of insulin and manganese on glucose oxidation was seen when Mn²⁺ was addedin vitro. The flux of glucose through the pentose phosphate pathway and glycolysis was significantly decreased in high fat fed animals. Although thein vitro addition of Mn²⁺ was additive with insulin when¹⁴CO2 was measured from control animals, it was found neither in young diabetic animals (6–8 weeks old) nor in the old (16 weeks old). Both insulin and manganese caused an increased oxidation of carbon-1 of glucose and an increase of its incorporation into¹⁴C-lipids in the young control animals; the additive effect of insulin and manganese suggests separate site of action. This effect was decreased in fat fed animals, diabetic animals and old animals. Manganese alone was found to decrease glycerol in both the control and diabetic adipose tissue inin vitro incubations. The results of the effects of glucose oxidation, lipogenesis, and glycerol release in adipose tissue of control and diabetic animals of different ages are presented together with the effect of manganese on adipose tissue from high fat milk diet fed animals.     

Studies on blood-brain barrier permeability after microwave-radiation

Summary Since the reported alterations of permeability of the blood-brain barrier by microwave radiation have implications for safety considerations in man, studies were conducted to replicate some of the initial investigations. No transfer of parenterally-administered fluorescein across the blood-brain barrier of rats after 30 min of 1.2-GHz radiation at power densities from 2–75 mW/cm² was noted. Increased fluorescein uptake was seen only when the rats were made hyperthermic in a warm-air environment. Similarly, no increase of brain uptake of¹⁴C-mannitol using the Oldendorf dual isotope technique was seen as a result of exposure to pulsed 1.3-GHz radiation at peak power densities up to 20 mW/cm², or in the continuous wave mode from 0.1–50 mW/cm². An attempt to alter the permeability of the blood-brain barrier for serotonin with microwave radiation was unsuccessful. From these studies it would appear that the brain must be made hyperthermic for changes in permeability of the barrier induced by microwave radiation to occur.The research reported in this paper was conducted by personnel of the Radiation Sciences Division, USAF School of Aerospace Medicine, Brooks AFB, Tex. 78235. The animals involved in the study were procured, maintained, and used in accordance with the Animal Welfare Act of 1970 and the Guide for the Care und Use of Laboratory Animals prepared by the Institute of Laboratory Animal Resources, National Research Council