Synthesis Of Serum Proteins Research Articles

Haematological and serum biochemical indices of growing female pigs fed dietary Spondias mombin leaf meal

Blood serves as physiological indicator of assessing quality of feed offered to an animal, their health status and body functionality. An experiment was carried out to evaluate the haematological and serum biochemical profiles of growing female pigs fed diets with varying levels of Spondias mombin leaf meal (SMLM). Sixteen growing female pigs weighing 10.5 to 14.0 kg were randomly allotted to four treatments supplemented maize feed ingredient with 0.0, 2.5, 5.0 and 7.5% in two phases of feeding with Spondias mombin leaf meal w/w in a Completely Randomized Design with four animals per treatment and replicated twice. Haematological and serum biochemical parameters were assessed. The results show that Packed Cell Volume (PCV), Red Blood Cell (RBC), Platelets and White Blood Cell (WBC) differentials were similar (p>0.05) in mean values, except for Hb which in T4 was significantly (p<0.05) different from other treatments in Hb (p<0.05). There were similarities in mean values of serum biochemical indices across treatments, except albumin and Alkaline Phosphatase (ALP) values in T3 indicating SMLM inclusion improves serum protein synthesis giving rise to more tissue formation. From this study, supplementation of SMLM up to 5.0% maize in growing female pigs’ diet have no deleterious effect on physiological health status and has potential to improve tissue development of animals.

Cadmium Toxicity: Insight into Sources, Toxicokinetics, and Effect on Vital Organs and Embryos

Background: Elemental cadmium, cadmium oxide, sulfide, and chloride are resurrected to the atmosphere from both natural and anthropogenic sources. cadmium concentrations in the blood, urine and kidney cortex are substantially higher in smokers compared to never smokers. Absorbed cadmium accumulates primly in the renal cortex and liver. The pancreas, thyroid, gall-bladder and testes can also include comparatively high concentrations. Objectives: The current review aimed to highlight on the effect of cadmium chloride on vital organs and embryos. Absorption of cadmium from the lungs is a major source of exposure between smokers and the occupationally exposed. Cadmium accumulates with age even a maximum level is arrived at about age 50. Metallothionein has been identified in the liver, kidneys, duodenum, urine and blood of animals. It has been postulated that metallothionein passes through the red cell membrane and is transformed to the kidney. Cadmium being a divalent cation is accumulated by transfers mechanisms developed for necessary metals. Cadmium may interact with zinc, iron, magnesium, manganese, calcium and selenium and cause their secondary disability and so break down metabolism, resulting in the final morphological and functional changes in many organs. Toxicity could result from cadmium (Cd2+) interacting with cellular ingredients until without get into the cell, but by interaction with receptors on their surface. Cadmium forms covalent and ionic bonds with atoms of sulfur, oxygen, and hydrogen present in the sulfhydryl groups, disulfide, carboxyl, imidazole or severally amino compounds present in the cells, causing significant disruption of their homeostasis. The principle target organelle of cadmium is the mitochondria. Symptoms of acute cadmium intoxicating usually appear after 24 hours are shortness of breath, general weakness, fever. It can also cause pulmonary edema, pneumonia and in acute cases, respiratory failure and death. Cadmium accumulates in the renal cortex and induces tubular damage. It may cause nephrotoxicity by procreating free radicals and/or by inducing necrosis, apoptosis and creatinine increase in plasma and urea increase in the serum. Exposure to cadmium can cause skeletal damage. Cadmium is attached to low bone mineralization, a high rate of fractures, increased osteoporosis and intense bone pain. It accumulates in the proximal tubular cells, it compresses cellular functions, which may result in reduced transformation among two forms of vitamin D3. This is probably to causes a decrease in calcium absorption and bone mineralization, which in turn may produce osteomalacia. The skeletal effects observed in young rats exposed to cadmium during the period of rapid skeletal growth and teratogenesis or skeletal effects recorded include sirenomelia (fused lower limbs), Amelia (absence of one or more limbs) and retard ossification of the sternum and ribs, dysplasia of facial bones and rear limbs and edema. Cadmium has been shown to be both embryotoxic and teratogenic in a different of animal species. It accumulates in human placenta and its concentration in cord blood increase with maternal exposure. Cadmium may be responsible for decreasing the volume of fetal capillaries in the terminal villi of the placenta. Furthermore, increasing the connective tissue about the fetal vessels changed synthesis of serum and amniotic fluid proteins and decreased expression of growth factors also may key a role in cadmium teratogenicity. Cadmium induces oxidative stress in many organisms at the cellular level, which may outcomes in physiological damage to various organs such as kidneys, liver, lung, pancreas, testes, placenta and bones Cadmium induces oxidative stress in many organisms at the cellular level, which may outcomes in physiological damage to various organs such as kidneys, liver, lung, pancreas, testes, placenta and bones. Reactive oxygen species reacting with polyunsaturated fatty acids of cell membranes initiate lipid peroxidation process that results in modulation of proteins, alteration in membrane components and this reason the loss of their impartiality and irrevocable damage. Conclusion: It can be concluded that symptoms of acute cadmium intoxicating are shortness of breath, general weakness, fever. pulmonary edema, and pneumonia. Cadmium accumulates with age in the different organs in the body and induces oxidative stress at the cellular level, which may outcomes in physiological damage to various organs such as kidneys, liver, lung, pancreas, testes, placenta and bones. Reactive oxygen species reacting with polyunsaturated fatty acids of cell membranes initiate lipid peroxidation process that results in modulation of proteins, alteration in membrane components and this reason the loss of their impartiality and irrevocable damage. Cadmium has been shown to be both embryotoxic and teratogenic in a different of animal species.

Changes in the Serum Protein Fractions in Goats after Treatment of Natural Gastrointestinal Parasite Infections.

Gastrointestinal parasitic infections in small ruminants belong to major health problems. The regulation of gastrointestinal infections in goats and the responses developed against them appear to be different from those observed in sheep. In the literature, there is a lack of data on the effect of gastrointestinal parasitic infections on the serum protein profile in goats. Therefore, the aim of the study was to determine the electrophoretic pattern of serum proteins in goats naturally infected with gastrointestinal parasites and to compare the changes in the total serum proteins and serum protein fractions (albumin and α1-, α2-, β-, and γ-globulins) obtained after antihelminthic treatment. Eight adult female goats of the white shorthaired breed from a small dairy goat farm at the age ranging between 3 and 5 years with average body weight 35.4 ± 3.2 kg and body condition score (BCS) from 1.5 to 2.5 were used in the study. The serum proteins in goats were separated into five fractions: albumin, α1- and α2-globulins, and β- and γ-globulins. Significant changes after treatment were found in the relative concentrations of albumin (P < 0.01) and α2- (P < 0.05), β- (P < 0.001), and γ-globulins (P < 0.01), as well as albumin/globulin ratio (P < 0.01). The mean concentration of total serum proteins was, after the antiparasitic treatment, significantly higher. Among the globulin fractions, the γ-globulin fraction contributed most significantly to these changes (P < 0.001). The results presented in the study suggest a significant effect of antiparasitic treatment in goats on the synthesis of blood serum proteins and on the changes of the proportion of serum protein fractions.

Hepatocyte cultures: From collagen gel sandwiches to microfluidic devices with integrated biosensors.

Hepatocytes are parenchymal cells of the liver responsible for drug detoxification, urea and bile production, serum protein synthesis, and glucose homeostasis. Hepatocytes are widely used for drug toxicity studies in bioartificial liver devices and for cell-based liver therapies. Because hepatocytes are highly differentiated cells residing in a complex microenvironment in vivo, they tend to lose hepatic phenotype and function in vitro. This paper first reviews traditional culture approaches used to rescue hepatic function in vitro and then discusses the benefits of emerging microfluidic-based culture approaches. We conclude by reviewing integration of hepatocyte cultures with bioanalytical or sensing approaches.

The Anti-depression Effect of Angelicae Sinensis Radix Is Related to the Pharmacological Activity of Modulating the Hematological Anomalies.

Angelicae Sinensis Radix (AS), a well-known herb in traditional Chinese medicine (TCM), has been wildly used for replenishing the blood and promoting circulation, in Asia for thousands of years. It has been confirmed that AS also possesses the pharmacological activity of anti-depression. At the same time, recent studies suggested that depression is associated with anemia, and depression could be ameliorated via modulating the blood system. However, it is still unknown whether the anti-depression effect of AS is related to its pharmacological activity of modulating the blood system. In the current study, hematological examination and metabonomic techniques were performed to explore potential anti-depression mechanisms of AS, related to the function of modulating the blood system in a chronic unpredictable mild stress (CUMS) model. The results demonstrated that AS could significantly improve CUMS-induced depressive symptom, hematological anomalies, and hypoxia symptoms. The analysis of metabonomics demonstrated that 26 potential biomarkers in depression could be regulated by the administration of AS. Among them, eight biomarkers participate in the metabolic pathways of amino acid and sphingolipid, and energy metabolism could also be regulated in an anemia model through the administration of AS, as reported in previous literatures. Further results proved that AS modulated energy metabolism in depression through the inhibition of the expression of pyruvate dehydrogenase lipoamide kinase isozyme 1 (PDK-1) and lactate dehydrogenase A (LDHA). These results suggested that the modulation of the blood system was involved in the anti-depression effect of AS. The mechanism may be associated with the promotion of the body’s energy metabolism, the stabilization of cell membranes, the promotion of serum protein synthesis, and the enhancement of immunity.

Oral Supplementation Using Gamma-Aminobutyric Acid and Whey Protein Improves Whole Body Fat-Free Mass in Men After Resistance Training.

BackgroundOral gamma-aminobutyric acid (GABA) supplementation increases growth hormone (GH) serum levels and protein synthesis. Therefore, post-exercise supplementation using GABA and protein may help enhance training-induced muscle hypertrophy. We evaluated whether GABA with whey protein enhanced muscular hypertrophy in men after progressive resistance training.MethodsTwenty-one healthy men (26 - 48 years) were randomized to receive whey protein (WP; 10 g) or whey protein + GABA (WP + GABA; 10 g + 100 mg) daily for 12 weeks. Both groups performed resistance training twice per week (three sets of 12 repetitions at 60% of maximal strength; leg press, leg extension, leg curl, chest press, and pull down). Body composition was assessed using dual-energy X-ray absorptiometry.ResultsIn the WP + GABA group, resting plasma GH concentrations were significantly elevated at 4 and 8 weeks, compared to baseline. However, resting plasma GH concentrations in the WP group were only significantly elevated at 8 weeks. After 12 weeks, the WP + GABA group exhibited significantly greater increase in whole body fat-free mass than the WP group.ConclusionsThe GABA and whey protein combination was more effective for increasing whole body fat-free mass; daily GABA supplementation may help enhance exercise-induced muscle hypertrophy.

Measurement of Albumin Secretion as Functionality Test in Primary Hepatocyte Cultures.

One of the most important functions of hepatocytes is the synthesis of serum proteins, more specifically of serum albumin. Albumin secretion in serum is essential, since it maintains the oncotic pressure in the body. Measurement of albumin secretion is used as a liver function test to indicate potential liver injury and liver pathology. In this chapter, a protocol for the measurement of albumin secretion in the supernatant of cultured rat hepatocytes is described. The procedure relies on an enzyme-linked immunosorbent assay allowing rat albumin to be quantitatively measured.

Ternary ZnO/ZnS/γ-Fe 2O 3 hollow sphere with surface hole: Microwave-enhanced rapid synthesis, bifunctional property, and immobilization of serum protein

High-yield ternary ZnO/ZnS/γ-Fe 2O 3 hollow spheres with an average size of 600 nm were prepared rapidly via a microwave-irradiation route; the reaction period is as short as 30 min. The hollow spheres are composed of numerous ZnO, ZnS, and γ-Fe 2O 3 crystallites and the mechanism responsible for their formation is Ostwald ripening. Every hollow sphere has a distinct surface hole with the pore size of tens of nanometers or hundreds of nanometers. The characterizations of the physical properties of the hollow spheres reveal that they have integrated the superparamagnetism of γ-Fe 2O 3 and the photoluminescence property of ZnO and ZnS. Furthermore, since the pore size of the hole on the surfaces of the hollow spheres is large enough to accommodate bovine serum albumin (BSA) molecule with a giant size, the hollow spheres can not only adsorb BSA with their external surfaces but also trap the proteins in their interiors. Per gram hollow sphere can immobilize 178.6 ± 34.5 mg of BSA when they are interacted with 600 μg/mL BSA solution for 24 h. The investigations on the releasing behavior of BSA by the hollow spheres reveal that BSA molecules on the external surfaces of the spheres are desorbed quickly, while those confined in the interiors of the spheres are released smoothly and the process is sustained over 10 days. The accumulative releasing percentage of BSA is about 51% in 10 days.

Mechanical Dissociation of Swine Liver to Produce Organoid Units for Tissue Engineering and In Vitro Disease Modeling

The complex intricate architecture of the liver is crucial to hepatic function. Standard protocols used for enzymatic digestion to isolate hepatocytes destroy tissue structure and result in significant loss of synthetic, metabolic, and detoxification processes. We describe a process using mechanical dissociation to generate hepatic organoids with preserved intrinsic tissue architecture from swine liver. Oxygen-supplemented perfusion culture better preserved organoid viability, morphology, serum protein synthesis, and urea production, compared with standard and oxygen-supplemented static culture. Hepatic organoids offer an alternative source for hepatic assist devices, engineered liver, disease modeling, and xenobiotic testing.

Progress toward liver‐based gene therapy

The liver is involved in the synthesis of serum proteins, regulation of metabolism and maintenance of homeostasis and provides a variety of opportunities for gene therapy. The enriched vasculature and blood circulation, fenestrated endothelium, abundant receptors on the plasma membranes of the liver cells, and effective transcription and translation machineries in the hepatocytes are some unique features that have been explored for delivery, and functional analysis, of genetic sequences in the liver. Both viral and non-viral methods have been developed for effective gene delivery and liver-based gene therapy. This review describes the fundamentals of gene delivery, and the preclinical and clinical progress that has been made toward gene therapy using the liver as a target.

Characteristics of lipid metabolism under different urinary protein excretion in children with primary nephrotic syndrome

Hyperlipidemia (HLP), a common complication, is very prevalent in children with primary nephrotic syndrome (PNS). HLP not only significantly increases the cardiovascular risk in adulthood, but also accelerates the progression of renal disease. Proteinuria as the most important pathophysiological change can reduce serum colloid osmotic pressure, which leads to an increase in the synthesis of serum proteins including lipoproteins in the liver for export to the serum. Thus, the severity of lipid abnormalities may correlate with the degree of proteinuria. A total of 378 children with PNS were divided into three groups according to their urinary protein excretion (UPE), group A (50 mg/kg/d ≤ proteinuria <100 mg/kg/d, 125 cases), group B (100 mg/kg/d ≤ proteinuria <200 mg/kg/d, 132 cases) and group C (proteinuria ≥200 mg/kg/d, 121cases). In addition, 200 healthy volunteers with neither allergic nor renal disease between 3 and 14 years of age were recruited as the control group. Fasting serum levels of lipoprotein (a) [Lp(a)], total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), apolipoprotein A1 (apoA1), apoB, and albumin (Alb) were measured. Serum low density lipoprotein cholesterol (LDL-C) was calculated by the Friedewald formula. As expected, when all patients were compared with healthy children in this study, UPE and the serum concentrations of Lp(a), TC, TG, HDL-C, LDL-C, and apoB were higher in the PNS than in the control group (p<0.01), whereas for apoA1/B ratio the opposite was observed (p<0.01). Furthermore, patients in group C exhibited significantly higher Lp(a), TC, TG, LDL-C, and apoB concentrations than those in group A or B (p<0.01), whereas for apoA1/B ratio the opposite was found (p<0.01). The increase in serum lipids was accompanied by a significant augmented UPE in all patients (p<0.05). More specifically, positive correlations were observed between serum levels of TC (r=+0.80, p<0.01), HDL (r=+0.49, p<0.01), LDL (r=+0.79, p<0.01), ApoB (r=+0.62, p<0.01) and log proteinuria in group B; additionally, a negative correlation was observed between apoA1/B ratio and log proteinuria in group B (r=−0.38, p<0.01). However, no correlation of serum lipid profiles with UPE was determined in group A and C, respectively (p>0.05). Serum Alb was negatively correlated with Lp(a) (r=−0.96, p<0.01), TC (r=−0.78, p<0.01), TG (r=−0.78, p<0.01), LDL-C (r=−0.88, p<0.01), apoA1 (r=−0.26, p<0.01), and apoB (r=−0.71, p<0.01), while positively correlated with apoA1/B (r=+0.27, p<0.01) in all nephrotic children. Furthermore, no correlation existed between serum lipid profiles and Alb in group A, B and C, respectively (p>0.05). In Conclusion, secondary dyslipidemia in children with PNS is in parallel with the degree of UPE. There are diverse characteristics of lipid metabolism under different UPE. As for the patients with medium-UPE, positive correlation between serum lipids and proteinuria is presented.

In vitro analysis of a hepatic device with intrinsic microvascular-based channels

A novel microfluidics-based bilayer device with a discrete parenchymal chamber modeled upon hepatic organ architecture is described. The microfluidics network was designed using computational models to provide appropriate flow behavior based on physiological data from human microvasculature. Patterned silicon wafer molds were used to generate films with the vascular-based microfluidics network design and parenchymal chamber by soft lithography. The assembled device harbors hepatocytes behind a nanoporous membrane that permits transport of metabolites and small proteins while protecting them from the effects of shear stress. The device can sustain both human hepatoma cells and primary rat hepatocytes by continuous in vitro perfusion of medium, allowing proliferation and maintaining hepatic functions such as serum protein synthesis and metabolism. The design and fabrication processes are scalable, enabling the device concept to serve as both a platform technology for drug discovery and toxicity, and for the continuing development of an improved liver-assist device.

Macromolecular Contrast Agents for Optical Imaging of Tumors: Comparison of Indotricarbocyanine-labeled Human Serum Albumin and Transferrin¶

Macromolecules accumulate in solid tumors and can thus be used as carriers for the delivery of attached contrast agents to tumors. We report the synthesis and use of serum protein–dye conjugates consisting of transferrin (Tf) or human serum albumin (HSA) and an indotricarbocyanine (ITCC) derivative as contrast agents for the optical imaging of tumors. The compounds were characterized with respect to their photophysical properties and tested in vitro for their ability to bind to tumor cells and in vivo for their potential to delineate experimental tumors. In contrast to HAS-ITTC, Tf-ITCC showed receptor-mediated uptake by HT29 human colon cancer cells in vitro. After intravenous injection into HT29 tumor-bearing nude mice both compounds induced increased fluorescence contrast of tumors in vivo. After 24 h the contrast between tumor and normal tissue was significantly higher for Tf-ITCC than for HAS-ITCC. Dye-induced fluorescence was found to be predominantly located in perinecrotic areas of the tumor. Furthermore, Tf-ITCC produced fluorescence of viable tumor cells, whereas HAS-ITCC fluorescence was recorded along connective tissue. We conclude that ITCC-labeled Tf and HSA can serve as macromolecular contrast agents for the optical imaging of tumors, with Tf-ITCC showing higher efficiency.

Protein turnover and sensory traits of longissimus muscle from implanted and nonimplanted heifers.

Primary bovine muscle cell culture studies were conducted to determine whether implanting heifers had a direct effect on in vitro protein synthesis and degradation and to determine the effect of implanting heifers on longissimus muscle palatability. Feedlot heifers (n = 96) were administered one of six implant regimens to characterize their effect on in vitro amino acid uptake and protein degradation. Treatments consisted of: 1) a nonimplanted control (NI/NI); 2) no implant on d 1 and Revalor-H administered on d 84 of the experiment (NI/Rev); 3) Revalor-H on d 1, but no implant given at d 84 (Rev/NI); 4) Revalor-H administered on d 1 and d 84 (Rev/Rev); 5) Revalor-IH administered on d 1 and Revalor-H at d 84 (RIH/Rev); and 6) Synovex-H given at d 1 and Revalor-H administered at d 84 (Syn/Rev). Blood and longissimus lumborum muscle were collected 20 min postmortem, and serum and muscle extracts were incubated with primary bovine muscle cells. Implant treatments had minimal effects on shear force and sensory traits; however, steaks from Rev/Rev heifers were 0.31 kg more tender (P < 0.05) than steaks from NI/NI heifers. Serum protein synthesis and degradation were not affected (P > 0.10) by any implant treatment. When primary bovine muscle cells were treated with muscle extract, amino acid uptake was greater for heifers implanted with Rev/ Rev than for the average of all other treatments (P < 0.01). The Rev/Rev implant regimen also increased (P < 0.05) amino acid uptake compared with heifers treated with RIH/Rev, Syn/Rev, NI/NI, NI/Rev, or Rev/NI. Cellular protein degradation of the muscle cell culture treated with muscle extract tended (P < 0.10) to be higher in NI/NI-treated cells compared with the average of all implant treatments. In addition, cells treated with muscle extract from heifers implanted with Rev/Rev had lower (P < 0.05) protein degradation than the NI/NI control heifers. These results indicate that anabolic implant strategies can directly affect both muscle protein synthesis and degradation via effects that seem to be more autocrine than paracrine in nature.

Polysomal ribonuclease 1 exists in a latent form on polysomes prior to estrogen activation of mRNA decay.

Estrogen induces a global change in the translation profile of Xenopus hepatocytes, replacing serum protein synthesis with production of the yolk protein precursor vitellogenin. This is accomplished by the coordinate destabilization of serum protein mRNAs and the transcriptional induction and subsequent stabilization of vitellogenin mRNA. Previous work identified an endonuclease activity whose appearance on polysomes correlated with the disappearance of serum protein mRNAs. This enzyme, polysomal ribonuclease 1 (PMR1), is a novel member of the peroxidase gene family. The current study examined the association of PMR1 with its mRNA targets on polysomes and mRNPs. The highest amount of polysome-bound PMR1 was observed prior to estrogen induction of mRNA decay. Its distribution on sucrose density gradients matched the absorbance profile of polysome-bound mRNA, suggesting that PMR1 forms a latent complex with mRNA. Following dissociation with EDTA the 62 kDa PMR1 sedimented with a larger complex of >670 kDa. Estrogen induces a 22-fold increase in unit enzymatic activity of polysome-bound PMR1, and a time-dependent loss of PMR1 from polysomes in a manner that mirrors the disappearance of albumin mRNA. These data suggest that the key step in the extensive estrogen-induced change in mRNA decay in Xenopus liver is activation of a latent mRNA endonuclease associated with its target mRNA.

PENGARUH EKSTRAK AKAR GINGSENG JAWA DALAM BERBAGAI KADAR TERHADAP SINTESIS PROTEIN SERUM TIKUS

The aim of this research was identifying the stimulation serum protein synthesis following the treatment of Java Gingseng (Talinum paniculatum Gaertn) root extract in varying concentration. This research used 12 male rats of 8-9 weeks, divided into 4 groups. First group as control, second group to five group were administratered with Java gingseng root extract equal with 3.5, 10.5, and 21 mg dry roots / 100g BW / day per oral for 14 days. Blood serum was collected for electrophoresized at the end of treatment. Electrophoresis was conducted with 7 percent SDS-PAGE method. Gel stained in 0.25 percent Coomasie Brilliant Blue and it followed densitometry on band in gel to comparing those quantitatively. The result show the effect on the level of serum protein whose MW 29. Highest stimulation of protein synthesis (2.2 times than control) have been reached at treatment with extract that equal with 3,5 mg dry root/ 100 g BW/ day. Increasing Java gingseng root extract was not followed by increasing of stimulation protein synthesis level.

Proteins of rat serum V: adjuvant arthritis and its modulation by nonsteroidal anti-inflammatory drugs.

The effect of adjuvant arthritis (AA) on the pattern of rat serum proteins includes the upregulation of haptoglobin, orosomucoid, alpha2-macroglobulin, serine protease inhibitor-3, thiostatin, alpha1-antitrypsin, C-reactive protein, and the downregulation of kallikrein-binding protein, alpha1-inhibitor III, apolipoprotein A-I, alpha2-HS-glycoprotein, albumin, apolipoprotein A-IV, transthyretin and transferrin. Minor changes (+/- 20%) are observed for Gc-globulin, ceruloplasmin, and alpha1-macroglobulin. AA thus grossly resembles the acute inflammatory response elicited by the injection of turpentine, although the changes in the levels of negative acute-phase proteins (APP) are smaller in acute inflammation. Indomethacine and ibuprofen inhibit the effects of arthritis on the synthesis of rat serum proteins in different ways: The former is, on average, three times as effective as the latter. Each drug interferes differently with different proteins. In animals without AA, both nonsteroidal anti-inflammatory drugs (NSAID) mimic the inflammatory pattern to a certain extent, with more effect on the negative than on the positive APPs. Overall, the shifts in serum protein levels parallel changes in inflammatory parameters such as joint swelling and serum interleukin-6 (IL-6) activity. Protein quantitation after two-dimensional electrophoresis (2-DE) reveals some effects of the drugs per se which escape detection by other routine tests.

The risk of surgery in patients with liver disease.

Because of the multiple functions of the liver—synthesis of most serum proteins, metabolism of nutrients and drugs, excretion and detoxification of endogenous toxins and exogenous agents, and filtering of portal venous blood—assessing the risk of surgery in patients with liver disease is a challenging endeavor. Any or all of the functions of the liver may be impaired in patients with liver disease.1,2 As a result, the pharmacokinetic parameters of anesthetics, muscle relaxants, analgesics, and sedatives can be affected by changes in binding to plasma proteins, detoxification, and excretion; bleeding risk can be increased because of coagulopathy; and susceptibility to infection can be increased because of altered functioning of hepatic reticuloendothelial cells and other changes in the immune system as well as portal hypertension. Perhaps most importantly, a diseased liver is particularly susceptible to the hemodynamic changes that accompany surgery. On the other hand, operative risk is probably not increased in the vast majority of patients with liver disease, including most of those with chronic hepatitis C, in whom liver function is preserved. Mild elevations of serum aminotransferase, alkaline phosphatase, or bilirubin levels are frequent after surgical procedures, whether performed under general, spinal, or epidural anesthesia.3,4 In patients without preexisting liver disease, these perturbations are usually transient, of no clinical significance, and often unnoticed. Clinically important hepatic dysfunction is more likely to occur in patients with preexisting liver disease. This review will consider the factors that contribute to perioperative hepatic dysfunction and prediction of operative risk in patients with liver disease undergoing surgery other than liver transplantation.

Serum Deprivation and Protein Synthesis Inhibition Induce Two Different Apoptotic Processes in N18 Neuroblastoma Cells

N18 are murine neuroblastoma cells that underwent cell death upon serum deprivation or inhibition of protein synthesis by means of cycloheximide (CHX). In both cases, an ultrastructural morphology and an internucleosomal pattern of DNA fragmentation typical of apoptosis were found. However, electron microscopy revealed abundant lipid vesicles in the cytoplasm of CHX-treated cells that were not found in their serum-deprived counterparts. In addition, when both types of apoptotic cells were compared by means of flow cytometry and chromatin staining with propidium iodide, the former showed consistently less fluorescence than the latter. Therefore, in N18 cells, both apoptotic processes seemed to differ at a structural level. At a functional level, we found that apoptosis was blocked by the protease inhibitor TLCK in CHX-treated but not in serum-deprived cells. On the other hand, we generated N18 clones that overexpressed Bcl-2 protein. After a period of 48 h we found that identical levels of Bcl-2 protein were able to block apoptosis in serum-deprived but not in CHX-treated cells. In conclusion, two different biochemical pathways leading to apoptosis seem to coexist in N18 neuroblastoma cells.

An attempt to add biological functions by genetic engineering in order to produce high-performance bioreactor cells for hybrid artificial liver: transfection of glutamine synthetase into Chinese hamster ovary (CHO) cell.

In the course of immortalization, hepatocyte cell lines lose their original differentiated functions, such as ammonia removal and urea formation, drug metabolism, serum protein synthesis, etc. (Enosawa et al., Cell Transplant. 5:S39-S40; 1996). With the aim of adding lost or deficient functions and producing cell lines for the bioreactor of a hybrid artificial liver, rat glutamine synthetase (GS) gene was transfected into Chinese hamster ovary (CHO) cells, because it is able to lower the ammonia level. The GS gene-inserted pSV2 plasmid was transfected into the CHO-K1 line by electroporation. Transfected CHO (GS-CHO) cells were cultured in a glutamine-free medium containing ammonia, glutamic acid, and the GS inhibitor methionine sulfoximine (MSX). The MSX concentration was increased stepwise from 25 mumol/L to 1600 mumol/L to amplify the GS gene. In several GS-CHO sublines resistant to 300-1600 mumol/L of MSX, the specific activities of GS were increased from 0.2 x 10(4) to 1.7-2.9 x 10(4) unit/10(6) cells. When the amplified GS-CHO cells were cultured in the ammonia-containing medium, a slow but steady decrease of the ammonia level was observed when the level was high. Finally, the prospect of genetically modulated cells for bioreactors in the hybrid artificial liver is discussed.