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1.

Objective

Glucose conversion into disaccharides was performed with β-glucosidases from Prunus dulcis (β-Pd), Aspergillus niger (β-An) and A. awamori (β-Aa), in reactions containing initial glucose of 700 and 900 g l?1.

Results

The reactions’ time courses were followed regarding glucose and product concentrations. In all cases, there was a predominant formation of gentiobiose over cellobiose and also of oligosaccharides with a higher molecular mass. For reactions containing 700 g glucose l?1, the final substrate conversions were 33, 38, and 23.5% for β-An, β-Aa, and β-Pd, respectively. The use of β-An yielded 103 g gentiobiose l?1 (15.5% yield), which is the highest reported for a fungal β-glucosidase. The increase in glucose concentration to 900 g l?1 resulted in a significant increase in disaccharide synthesis by β-Pd, reaching 128 g gentiobiose l?1 (15% yield), while for β-An and β-Aa, there was a shift toward the synthesis of higher oligosaccharides.

Conclusion

β-Pd and the fungal β-An and β-Aa β-glucosidases present quite dissimilar kinetics and selective properties regarding the synthesis of disaccharides; while β-Pd showed the highest productivity for gentiobiose synthesis, β-An presented the highest specificity.
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2.

Background

Diabetes mellitus is a group of metabolic diseases with increased blood glucose concentration as the main symptom. This can be caused by a relative or a total lack of insulin which is produced by the β‐cells in the pancreatic islets of Langerhans. Recent experimental results indicate the relevance of the β‐cell cycle for the development of diabetes mellitus.

Methods

This paper introduces a mathematical model that connects the dynamics of glucose and insulin concentration with the β‐cell cycle. The interplay of glucose, insulin, and β‐cell cycle is described with a system of ordinary differential equations. The model and its development will be presented as well as its mathematical analysis. The latter investigates the steady states of the model and their stability.

Results

Our model shows the connection of glucose and insulin concentrations to the β‐cell cycle. In this way the important role of glucose as regulator of the cell cycle and the capability of the β‐cell mass to adapt to metabolic demands can be presented. Simulations of the model correspond to the qualitative behavior of the glucose‐insulin regulatory system showed in biological experiments.

Conclusions

This work focusses on modeling the physiological situation of the glucose‐insulin regulatory system with a detailed consideration of the β‐cell cycle. Furthermore, the presented model allows the simulation of pathological scenarios. Modification of different parameters results in simulation of either type 1 or type 2 diabetes.
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3.
The chemical structure of hydrothermally treated β-1,3–1,6-glucan from Aureobasidium pullulans was characterized using techniques such as gas chromatography/mass spectrometry (GC/MS) and nuclear magnetic resonance (NMR). The chemical shifts of anomeric carbons observed in the 13C-NMR spectra suggested the presence of single flexible chains of polysaccharide in the sample. β-1,3–1,6-Glucan from A. pullulans became water-soluble, with an average molecular weight of 128,000 Da after hydrothermal treatment, and the solubility in water was approximately 10% (w/w). Sample (3% w/v) was completely hydrolyzed to glucose by enzymatic reaction with Lysing enzymes from Trichoderma harzianum. Gentiobiose (Glcβ1 → 6Glc) and glucose were released as products during the reaction, and the maximum yield of gentiobiose was approximately 70% (w/w). The molar ratio of gentiobiose to glucose after 1 h reaction suggested that the sample is likely highly branched. Sample (3% w/v) was also hydrolyzed to glucose by Uskizyme from Trichoderma sp., indicating that it is very sensitive to enzymatic hydrolysis.  相似文献   

4.
In the course of taxonomic studies on saprobic microfungi from Spain, several slow-growing, dematiaceous hyphomycetes were isolated from soil, submerged plant material and river sediments. Sixteen of these strains were identified as members of the ascomycete order Chaetothyriales on the basis of morphology and DNA sequence analyses of the internal transcribed spacer region and partial large subunit ribosomal RNA gene. These included three novel species (Cladophialophora pseudocarrionii, Cyphellophora chlamydospora, and Rhinocladiella amoena) and five interesting, little-known or clinically-relevant species (Cyphellophora suttonii, Exophiala aquamarina, E. lacus, E. radicis, and Rhinocladiella similis). In addition, Exophiala oligosperma, an emerging opportunistic fungus, was found for the first time in an aquatic freshwater environment (river sediments). Cladophialophora pseudocarrionii resembles C. carrionii in the branching pattern of its conidial chains, but differs from the latter species in its inability to grow at 30 °C. Cyphellophora chlamydospora differs from other species of the genus in the absence of conidiation, producing only chlamydospores in vitro. Rhinocladiella amoena shows branched conidiophores similar to those of R. anceps, R. atrovirens, R. basitona and R. similis, but differs from them in conidial shape and size. The ex-type strain of Phialophora livistonae, included in the phylogenetic study, clustered with high statistical support with members of the genus Cyphellophora and is transferred to this genus.  相似文献   

5.
An axenic culture of a cyanobacterium (strain HPS) was raised from a field specimen of greenish planktonic floccose mass from a local lake, Ganga Sagar. Intense morphological examination and comparative morphological assessment with the genera Fischerella and Hapalosiphon and all the known strains of the genus Westiellopsis indicated that the strain HPS differed in morphology with the closely related strains in trichome arrangement, size of vegetative cells, heterocytes, monocytes, and number of rows of main filaments. There were also differences in the habitat preference, being aquatic rather than terrestrial or subaerial. Intense ecological characterization of the habitat was performed through measurements of important physicochemical characteristics. 16S rRNA gene-based identification and phylogenetic placement indicated conclusively that the strain HPS was different from the most closely related strain Westiellopsis prolifica SAG 16.93. Phylogenetic inferences drawn in between all the branched heterocytous forms and subsequent 16S-23S ITS analyses and folding of the secondary structures revealed an entirely new form that is unknown till now. Subsequent nifD and rbcL gene-based phylogenetic assessments indicated that strain HPS is phylogenetically different from all the other previously known species of true branching cyanobacteria, along with also pointing toward the huge database inconsistencies in case of true branched cyanobacteria. Assessment of morphological and ecological differences along with comprehensive phylogenetic evaluation indicated that the strain HPS is a new species of the genus Westiellopsis and the name being proposed is Westiellopsis ramosa sp. nov.  相似文献   

6.
Aerobic bacteria utilizing methane as the carbon and energy source do not use sugars as growth substrates but possess the gene coding for glucokinase (Glk), an enzyme converting glucose into glucose 6-phosphate. Here we demonstrate the functionality and properties of Glk from an obligate methanotroph Methylomicrobium alcaliphilum 20Z. The recombinant Glk obtained by heterologous expression in Escherichia coli was found to be close in biochemical properties to other prokaryotic Glks. The homodimeric enzyme (2 × 35 kDa) catalyzed ATP-dependent phosphorylation of glucose and glucosamine with nearly equal activity, being inhibited by ADP (K i = 2.34 mM) but not affected by glucose 6-phosphate. Chromosomal deletion of the glk gene resulted in a loss of Glk activity and retardation of growth as well as in a decrease of intracellular glycogen content. Inactivation of the genes encoding sucrose phosphate synthase or amylosucrase, the enzymes involved in glycogen biosynthesis via sucrose as intermediate, did not prevent glycogen accumulation. In silico analysis revealed glk orthologs predominantly in methanotrophs harboring glycogen synthase genes. The data obtained suggested that Glk is implicated in the regulation of glycogen biosynthesis/degradation in an obligate methanotroph.  相似文献   

7.
On the basis of a two-component (two-fluid) hydrodynamic model, it is shown that the probable phenomenon of solar core rotation with a velocity higher than the average velocity of global rotation of the Sun, discovered by the SOHO mission, can be related to fast solid-body rotation of the light hydrogen component of the solar plasma, which is caused by thermonuclear fusion of hydrogen into helium inside the hot dense solar core. Thermonuclear fusion of four protons into a helium nucleus (α-particle) creates a large free specific volume per unit particle due to the large difference between the densities of the solar plasma and nuclear matter. As a result, an efficient volumetric sink of one of the components of the solar substance—hydrogen—forms inside the solar core. Therefore, a steady-state radial proton flux converging to the center should exist inside the Sun, which maintains a constant concentration of hydrogen as it burns out in the solar core. It is demonstrated that such a converging flux of hydrogen plasma with the radial velocity v r (r) = ?βr creates a convective, v r ?v φ/?r, and a local Coriolis, v r v φ/r,φ nonlinear hydrodynamic forces in the solar plasma, rotating with the azimuthal velocity v φ. In the absence of dissipation, these forces should cause an exponential growth of the solid-body rotation velocity of the hydrogen component inside the solar core. However, friction between the hydrogen and helium components of the solar plasma due to Coulomb collisions of protons with α-particles results in a steady-state regime of rotation of the hydrogen component in the solar core with an angular velocity substantially exceeding the global rotational velocity of the Sun. It is suggested that the observed differential (liquid-like) rotation of the visible surface of the Sun (photosphere) with the maximum angular velocity at the equator is caused by sold-body rotation of the solar plasma in the radiation zone and strong turbulence in the tachocline layer, where the turbulent viscosity reaches its maximum value at the equator. There, the tachocline layer exerts the most efficient drag on the less dense outer layers of the solar plasma, which are slowed down due to the interaction with the ambient space plasma (solar wind).  相似文献   

8.
Cotton (Gossypium spp.), as the most important fiber and oilseed crop in the world, is extremely important for the industry. However, due to its indeterminate growth habit and complex branching system, massive labor costs are needed for shoot apex removal and branch pruning during cotton production. Therefore, it is very important to explore branch-controlling genes and genetically modify the branch architecture of cotton. Strigolactones (SLs) are a novel class of plant hormone that inhibit the outgrowth of lateral branches. To elucidate the role of SLs in branch development of cotton, we cloned and characterized GhMAX2a and GhMAX2b from tetraploid upland cotton (Gossypium hirsutum), the orthologs of Arabidopsis MAX2, rice D3, and petunia RMS4. GhMAX2a/2b was ubiquitously expressed in all tested tissues of cotton, with relatively higher expression levels in leaves and lateral buds. Subcellular localization assay showed that the GhMAX2-GFP fusion protein localized to the nucleus. Both GhMAX2a and GhMAX2b can fully rescue the dwarfed and highly branched phenotypes of the Arabidopsis max2-1 mutant, indicating that GhMAX2s have conserved functions with that of AtMAX2. The cotton GhMAX2b interacted with Arabidopsis Skp1-like 1 (ASK1) proteins in vitro which was further confirmed in the Arabidopsis protoplasts using the co-immunoprecipitation assay, indicating that GhMAX2b probably functions through forming an SCF E3 complex with Skp and other proteins in the Arabidopsis. These results suggest that the cotton GhMAX2s encode functional MAX2 that can inhibit the shoot lateral branching. Further functional analysis of GhMAX2s in determining cotton branch architecture and yield is underway.  相似文献   

9.
Six new natural compounds were isolated from two Far Eastern starfish species, Henricia aspera and H. tumida, collected in the Sea of Okhotsk. Two new glycosylated steroid polyols were obtained from H. aspera: asperoside A and asperoside B, which were shown to be (20R,24R, 25S)-3-O-(2,3-di-O-methyl-β -D-xylopyranosyl)-24-methyl-5α-cholest-4-ene-3β, 6β,8,15α,16β,26-hexaol and (20R, 24R,25S,22E)-3-O-(2,4-di-O-methyl-β-D-xylopyranosyl)-24-methyl-5α-cholest-22-ene-3β,4β,6β,8,15α,26-hexaol, respectively. Two other glycosylated polyols, tumidoside A, with the structure elucidated as (20R, 22E)-3-O-(2,4-di-O-methyl-β -D-xylopyranosyl)-26,27-dinor-24-methyl-5α-cholest-22-ene-3β,4β,6β,8,15α,25-hexaol, and tumidoside B, whose structure was elucidated as (20R,24S)-3-O-(2,3-di-O-methyl-β-D-xylopyranosyl)-5α-cholestan-3β,4β,6β,8,15α,24-hexaol, were isolated from the two starfish species. (20R, 24S)-5α-Cholestan-3β,6β,15α,24-tetraol and (20R, 24S)-5α-cholestan-3β,6β,8,15α,24-pentaol were identified only in H. tumida. The known monoglycosides henricioside H1 and laeviuscolosides H and G were also identified in both species.  相似文献   

10.
Dynamic light scattering was used to study the interaction of phosphorylase kinase (PhK) and glycogen phos-phorylase b (Phb) from rabbit skeletal muscle with glycogen under molecular crowding conditions arising from the presence of 1 M trimethylamine N-oxide and at physiological ionic strength. The mean value of hydrodynamic radius of the initial glycogen particles was 52 nm. Crowding stimulated Phb and PhK combined binding on glycogen particles. Two-stage character of PhK binding to glycogen particles containing adsorbed Phb was found in the presence of the crowding agent. At the initial stage, limited size particles with hydrodynamic radius of ~220 nm are formed, whereas the second stage is accompanied by linear growth of hydrodynamic radius. Flavin adenine dinucleotide (FAD) selectively inhibited PhK binding at the second stage. The data indicate that in the first stage Phb is involved in PhK binding by glycogen particles containing adsorbed Phb, whereas PhK binding in the second stage does not involve Phb.  相似文献   

11.

Background

Platelet-derived growth factor receptor α (PDGFRα) expression is increased in activated hepatic stellate cells (HSCs) in cirrhotic liver, while normal hepatocytes express PDGFRα at a negligible level. However, cancerous hepatocytes may show upregulation of PDGFRα, and hepatocellular carcinoma is preceded by chronic liver injury. The role of PDGFRα in non-cancerous hepatocytes and liver fibrosis is unclear. We hypothesized that upon liver injury, PDGFRα in insulted hepatocytes contributes to liver fibrosis by facilitating intercellular crosstalk between hepatocytes and HSCs.

Methods

Hepatocytes were isolated from normal and thioacetamide (TAA)-induced cirrhotic livers for assessment of PDGFRα expression. Conditional knock-out (KO) C57BL/6 mice, in which PDGFRα was selectively deleted in hepatocytes, were generated. Liver fibrosis was induced by injecting TAA for 8?weeks. Hep3B cells were transfected with a small interfering RNA (siRNA) (PDGFRα or control) and co-cultured with LX2 cells.

Results

PDGFRα expression was increased in hepatocytes from fibrotic livers compared to normal livers. Conditional PDGFRα KO mice had attenuated TAA-induced liver fibrosis with decreased HSC activation and proliferation. Immunoblot analyses revealed decreased expression of phospho-p44/42 MAPK in TAA-treated KO mice; these mice also showed almost complete suppression of the upregulation of mouse double minute 2. Although KO mice exhibited increased expression of transforming growth factor (TGF)-β and Smad2/3, this was compensated for by increased expression of inhibitory Smad7. LX2 cells co-cultured with PDGFRα siRNA-infected Hep3B cells showed decreased PDGFRα, α smooth muscle actin, collagen α1(I), TGFβ, and Smad2/3 expression. LX2/PDGFRα-deleted hepatocyte co-culture medium showed decreased PDGF-BB and PDGF-CC levels.

Conclusions

Deletion of PDGFRα in hepatocytes attenuated the upregulation of PDGFRα in HSCs after TAA treatment, resulting in decreased liver fibrosis and HSC activation. This suggests that in the event of chronic liver injury, PDGFRα in hepatocytes plays an important role in liver fibrosis by affecting PDGFRα expression in HSCs.
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12.
13.
Human amniotic membrane (HAM) due to its high biocompatibility, low immunogenicity, anti-microbial, anti-viral properties as well as the presence of growth factors has been used in various clinical applications. The growth factors play an important role in wound healing. The current study aimed to explore the effect of 15 kGy gamma radiation dose on selected growth factors and receptors mRNA present in HAM. Eight growth factors, namely, EGF, HGF, KGF, TGF-α, TGF-β1, TGF-β2, TGF-β3 and bFGF and two growth factor receptors, HGFR and KGFR were evaluated in this study. The total RNA was extracted and converted to complimentary DNA using commercial kits. Subsequently, the mRNA expressions of these growth factors were evaluated using real-time PCR and the results were statistically analyzed using REST-MCS software. This study confirmed the presence of these mRNA growth factors and receptors in fresh, glycerol cryopreserved and irradiated glycerol cryopreserved HAM. In glycerol cryopreserved HAM, the results showed up-regulation of HGF and bFGF and down-regulation of EGF, HGFR, KGF, KGFR, TGF-α, TGF-β1, TGF-β2 and TGF-β3 relative to the fresh HAM which acted as the control, whereas in irradiated glycerol cryopreserved HAM, the results showed up-regulation of EGF, HGF, KGF, KGFR, TGF-β1, TGF-β2 and TGF-β3 and down-regulation of HGFR, TGF-α and bFGF relative to the glycerol cryopreserved HAM which acted as the control. However, these mRNA expressions did not show any statistical significant difference compared to the control groups. This study concluded that a dose of 15 kGy of gamma radiation did not affect the mRNA expression for the growth factors’ and receptors’ in the glycerol cryopreserved HAM.  相似文献   

14.

Background

Insulin secreted by pancreatic islet β-cells is the principal regulating hormone of glucose metabolism and plays a key role in controlling glucose level in blood. Impairment of the pancreatic islet function may cause glucose to accumulate in blood, and result in diabetes mellitus. Recent studies have shown that mitochondrial dysfunction has a strong negative effect on insulin secretion.

Methods

In order to study the cause of dysfunction of pancreatic islets, a multiple cell model containing healthy and unhealthy cells is proposed based on an existing single cell model. A parameter that represents the function of mitochondria is modified for unhealthy cells. A 3-D hexagonal lattice structure is used to model the spatial differences among β-cells in a pancreatic islet. The β-cells in the model are connected through direct electrical connections between neighboring β-cells.

Results

The simulation results show that the low ratio of total mitochondrial volume over cytoplasm volume per β-cell is a main reason that causes some mitochondria to lose their function. The results also show that the overall insulin secretion will be seriously disrupted when more than 15% of the β-cells in pancreatic islets become unhealthy.

Conclusion

Analysis of the model shows that the insulin secretion can be reinstated by increasing the glucokinase level. This new discovery sheds light on antidiabetic medication.
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15.
Andrographolide (AP), a phytoconstituent of Andrographis paniculata is reported as a potent hepatoprotective agent. However, utility of this molecule is restricted due to its low aqueous solubility, gastric instability and hence low bioavailability. It was aimed to formulate and characterize AP-loaded, natural biopolymer stabilized, multilayered nano-hydrocolloid delivery system. Nanoemulsion (NE) was formulated using layer-by-layer (LbL) technology via electrostatic deposition of chitosan over alginate encrusted o/w NE by ultra-sonication. Improved transparency and stability of NE were observed with increasing sonication time. Best stability was obtained after 20 min sonication and particle size of the multilayered NE was measured in the range of 90.8–167.8 nm. Transmission electron microscopy confirmed the progressive layering of nanosized NE. Higher magnitude of zeta potential (i.e., 22.9 to 31.01 mV) confirmed higher stability and coating of alginate layer over NE surface for the period of 3 months. NE showed strategic release pattern when assessed in vitro in various simulated biological fluids of GIT in timed pattern. Multilayered NE showed significant modulation in liver function test (ALT, ALP, AST, TBIL, DBIL, and liver glycogen) and serum cytokines (TNF-α, IL-6, IL-10, and IL-β) when assessed in vivo in galactosamine-lipopolysaccharide intoxicated mice. In conclusion, the andrographolide engrained multi-layered NE enhanced the solubility, stability and henceforth assured the increased availability in simulated biological fluids. The in vivo study exhibited the significantly improved hepatoprotection by andrographolide when delivered in stable multi-layered NE carrier systems.  相似文献   

16.
17.
Acinetobacter baylyi ADP1 naturally produces wax esters that could be used as a raw material in industrial applications. We attempted to improve wax ester yield of A. baylyi ADP1 by removing rmlA, a gene involved in exopolysaccharide production. Growth rate, biomass formation and wax ester yield on 4-hydroxybenzoate were not affected, but the rmlA ? strain grew slower on acetate, while reaching similar biomass and wax ester yield. The rmlA ? cells had malformed shape and large size and grew poorly on glucose without expression of the gene for pyruvate kinase (pykF) from Escherichia coli. The pykF-expressing rmlA ? strain had similar growth rate, lowered biomass formation and improved wax ester production on glucose as compared to the wild-type strain expressing pykF. Cultivation of the pykF-expressing rmlA ? strain on an elevated glucose concentration in a medium supplemented with amino acids resulted in doubled molar wax ester yield and acetate production.  相似文献   

18.
Three new steroid glycosides (evasteriosides C, D, and E) along with six known compounds were isolated from two Pacific starfish of the genus Evasterias. Evasterioside C from E. retiferacollected from the Sea of Japan was identified as (20R, 22E)-3-O-(β-D-xylopyranosyl)-24-nor-5α-cholest-22-ene-3β,6β,15α,26-pentaol 26-sulfate sodium salt. The structures of evasteriosides D and E from E. echinosoma (collected from the Gulf of Shelichov, the Sea of Okhotsk) were established as (20R, 24S)-24-O-(β-D-glucopyranosyl)-5α-cholestane-3β,6α,8,15β,24-pentaol and (20R,24S)-3,24-di-O-(β-D-xylopyranosyl)-cholest-4-ene-3β,6β,8,15α,24-pentaol, respectively. In addition, the known compounds pycnopodiosides A and C, luridoside A, 5α-cholestane-3β,6α,8,15β,16β,26-hexaol. 5α-Cholestane-3β,6α,8,15β,24-pentaol 24-sulfate sodium saltand marthasterone sulfate sodium salt were identified in E. echinosoma. The structures of the isolated compounds were established on the basis of spectroscopic analyses, using 1D and 2D NMR techniques, mass spectrometry, and some chemical transformations.  相似文献   

19.
Increasing studies have shown protective effects of intermittent hypoxia on brain injury and heart ischemia. However, the effect of intermittent hypoxia on blood glucose metabolism, especially in diabetic conditions, is rarely observed. The aim of this study was to investigate whether intermittent hypoxia influences blood glucose metabolism in type 1 diabetic rats. Streptozotocin-induced diabetic adult rats and age-matched control rats were treated with intermittent hypoxia (at an altitude of 3 km, 4 h per day for 3 weeks) or normoxia as control. Fasting blood glucose, body weight, plasma fructosamine, plasma insulin, homeostasis model assessment of insulin resistance (HOMA-IR), pancreas β-cell mass, and hepatic and soleus glycogen were measured. Compared with diabetic rats before treatment, the level of fasting blood glucose in diabetic rats after normoxic treatment was increased (19.88?±?5.69 mmol/L vs. 14.79?±?5.84 mmol/L, p?<?0.05), while it was not different in diabetic rats after hypoxic treatment (13.14?±?5.77 mmol/L vs. 14.79?±?5.84 mmol/L, p?>?0.05). Meanwhile, fasting blood glucose in diabetic rats after hypoxic treatment was also lower than that in diabetic rats after normoxic treatment (13.14 ± 5.77 mmol/L vs. 19.88 ± 5.69 mmol/L, p<0.05). Plasma fructosamine in diabetic rats receiving intermittent hypoxia was significantly lower than that in diabetic rats receiving normoxia (1.28?±?0.11 vs. 1.39?±?0.11, p?<?0.05), while there were no significant changes in body weight, plasma insulin and β-cell mass. HOMA-IR in diabetic rats after hypoxic treatment was also lower compared with diabetic rats after normoxic treatment (3.48?±?0.48 vs. 3.86?±?0.42, p?<?0.05). Moreover, intermittent hypoxia showed effect on the increase of soleus glycogen but not hepatic glycogen. We conclude that intermittent hypoxia maintains glycemia in streptozotocin-induced diabetic rats and its regulation on muscular glycogenesis may play a role in the underlying mechanism.  相似文献   

20.
The red-toothed shrews (genus Sorex) are one of the smallest mammals. The amount of food they consume a day exceeds their own weight, and without food they can survive for only few hours. Representatives of this genus have extremely high metabolic rates. This study addressed the effect of 8-h fasting and 13-h refeeding on the body weight, blood glucose level, liver glycogen and lipid levels, and relative weight of inguinal and interscapular adipose tissues in the even-toothed shrews (S. isodon). Fasting led to a decrease in the body weight, blood glucose and liver glycogen levels. The relative weight of adipose tissue also decreased, while the liver lipid level increased significantly. After refeeding, blood glucose and liver glycogen levels were considerably higher than in control, while other parameters remained almost the same as in control. Physiological response to fasting develops in S. isodon quite rapidly, promoted by the high metabolic rate.  相似文献   

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