Alpha-fetoprotein Synthesis Research Articles

A simple, sensitive, label-free electrochemical immunosensor based on the chitosan-coated silver/cerium oxide (CS@Ag/CeO2) nanocomposites for the detection of alpha-fetoprotein (AFP)

Metal oxide-based sensors have the benefit of inexpensive, quick response, and high sensitivity in detecting specific biological species. In this article, a simple electrochemical immunosensor was fabricated using antibody-chitosan coated silver/cerium oxide (Ab-CS@Ag/CeO2) nanocomposites on a gold electrode for sensitive alpha-fetoprotein (AFP) diagnosis in human serum samples. Successfully synthesis of AFP antibody-CS@Ag/CeO2 conjugates was confirmed through Fourier transform infrared spectra of the prototype. The amine coupling bond chemistry was then used to immobilize the resultant conjugate on a gold electrode surface. It was observed that the interaction of the synthesized Ab-CS@Ag/CeO2 nanocomposites with AFP prevented an electron transfer and reduced the voltammetric Fe(CN)6 3−/4− peak current, which was proportional to the amount of AFP. The linear ranges of AFP concentration were found from 10−12–10−6 g.ml−1. The limit of detection was calculated using the calibration curve and came out to be 0.57 pg.ml−1. The designed label-free immunosensor successfully detected AFP in human serum samples. As a result, the resulting immunosensor is a promising sensor plate form for AFP detection and could be used in clinical bioanalysis.

URRENT CONCEPT OF THE STRUCTURAL AND FUNCTIONAL PROPERTIES OF ALFA-FETOPROTEIN AND THE POSSIBILITIES OF ITS CLINICAL APPLICATION

This paper was aimed to review the literature data from native and foreign sources accumulated for 40-years period of research of the features of the molecular structure, functions, production and application of human alpha-fetoprotein (AFP), which is known as one of the most studied and increasingly demanded proteins. Results of fundamental studies performed with the use of modern methods, including various types of electrophoresis, chromatography, electron microscopy and immunoassay, in order to characterize the principal physicochemical capacities and localization of free and bound forms of AFP, as well as polypeptide structure, heterogeneity and topography of AFP receptors are highlighted here. The data on the mechanisms of AFP synthesis, its conformational features, binding sites and intracellular metabolism are also presented. The concepts of physiological functions and mechanisms of AFP transport in an organism are presented. Data on AFP isolation from the natural primary products and its production by means of recombinant and synthetic methods are shown. This review also summarizes information on the current possibilities of clinical application of AFP and the prospects for its usage in anticancer therapy for targeted delivery of chemotherapy drugs, with emphasis on the description of the recent progress in this field.

Liquidus Tracking: Large scale preservation of encapsulated 3-D cell cultures using a vitrification machine

Currently, cryo-banking of multicellular structures such as organoids, especially in large volumes at clinical scale >1 L, remains elusive for reasons such as insufficient dehydration and cryoprotectant additive (CPA1) penetration, slow cooling and warming rates and devitrification processes. Here we introduce the concept of Liquidus Tracking (LT) using a semi-automated process for liquid volumes of up to 450 ml including 130 ml of alginate encapsulated liver cells (AELC) that archived controlled and reversible vitrification with minimized toxicity.First a CPA solution with optimal properties for LT was developed by employing different small scale test systems. Combining sugars such as glucose and raffinose with Me2SO improved post-exposure (at +0.5 °C) viabilities from 6% ±3.6 for Me2SO alone up to 58% ±6.1 and 65% ±14.2 respectively (p < 0.01). Other permeating CPAs (e.g. ethylene glycol, propylene glycol, methanol) were investigated as partial replacements for Me2SO. A mixture of Me2SO, ethylene glycol and glucose (ratio 4:2:1– termed LTdeg) supported glass-forming tendencies with appropriate low viscosities and toxicities required for LT. When running the full LT process, using Me2SO alone, no viable cells were recovered; using LTdeg, viable recoveries were improved to 40% ±8 (p<0.001%). Further refinements of improved mixing technique further improved recovery after LT. Recoveries of specific liver cell functions such as synthesis of albumin and alpha-fetoprotein (AFP) were retained in post thaw cultures.In summary: By developing a low-toxicity CPA solution of low viscosity (LTdeg) suitable for LT and by improving the stirring system, post-warming viability of AELC of up to 90% and a AFP secretion of 89% were reached. Results show that it may be possible to develop LT as a suitable cryogenic preservation process for different cell therapy products at large scale.

Synthesis of alpha-fetoprotein by human yolk sac tumor transplanted into nude mice.

All nude mice bearing primary or secondary transplants of a human yolk sac tumor exhibited a high serum alpha-fetoprotein level. This finding clearly indicates the synthesis of alpha-fetoprotein by the human yolk sac tumor and supports an idea that human yolk sac tumor simulates yolk sac endoderm not only morphologically but also functionally.

Impact of alginate type and bead diameter on mass transfers and the metabolic activities of encapsulated C3A cells in bioartificial liver applications

Liver-assist devices have been developed in the last few decades to support patients with liver failure on the road to recovery or transplantation. Fluidised bed bio-artificial livers--where liver cells are encapsulated within alginate beads--appear to be a valuable alternative to hollow fibre devices for improving mass transfers and enhancing treatment efficacy. This approach nevertheless deserves optimization in terms of bead production. The aim of this study was to investigate the impact of alginate type and of two bead diameters (1000 µm and 600 µm) on mass transfers within beads and on the biological functions of encapsulated C3A cells. After assessing the effect of the encapsulation process on bead quality, we investigated cell viability and metabolic activities (ammonia, albumin, alpha-fetoprotein synthesis and glucose consumption). They were successfully maintained over 48 h within fluidised bed bioreactors, independently of alginate type and bead diameter. Mass transfers were not significantly influenced by the latter parameters. Finally, suggestions are made for improving the entrapment process as a means of enhancing the treatment efficiency of the fluidised bed bioartificial liver.

Histopathology and immunohistochemistry of large hepatocellular carcinoma with undetectable or low serum levels of alpha-fetoprotein.

To explore the pathomorphological characteristics of large hepatocellular carcinoma (LHCC) with low serum alpha-fetoprotein (AFP) level. Specimens obtained from surgically resected LHCC with undetectable or low levels of serum AFP were fixed in formalin, embedded in paraffin, and prepared as serial sections. Routine hematoxylin and eosin as well as immunohistochemical stains (LSAB method) were used to test for expression of AFP, alpha-1-antitrypsin, epithelial membrane antigen, and vimentin. Some characteristics of the histopathological changes and immunohistochemical reactions of the cancerous tissues were observed under the light microscope. The majority of the cases (19/30) of LHCC with undetectable or low levels of serum AFP were of the clear-cell-type HCC, with 2 being positive for AFP expression at the periphery of the cytoplasm. The clear cell is the morphological manifestation of disturbance in glycogen and/or lipid metabolism of hepatoma cells. Such changes might be one of the factors hindering the synthesis of AFP and resulting in negative or low level serum AFP of the patient.

High Alpha-Fetoprotein Persistence after Orchiectomy. On a Case of Uncommon Etiology

Background The following report describes a case of inherited elevation of alpha-fetoprotein (AFP) in a young male suspected for testicular cancer. AFP shows very high values during fetal life. After birth the synthesis of AFP decreases dramatically, and only trace amounts are detected in the adult. From this age on, serum AFP can rise above normal in some diseases, e.g. liver disorders, and in some kind of tumors. A condition in which persistent high levels of AFP are found, named Hereditary Persistence of AFP [HPAFP), was first reported in 1983 by Ferguson-Smith, and then recorded in the literature only on eleven occasions till 2004. The occurrence of HPAFP may be underestimated. HPAFP can be easily confirmed by testing AFP levels in blood-related family members. Methods An elevated serum AFP (about 20 μg/mL) was found in a 27–year-old white man with an unremarkable medical history, who was concerned to have left testicular cancer. By our examination, his left testis was markedly reduced in size. β-HCG, LDH, and liver function were normal. Surgical inguinal exploration with testis and spermatic cord excision was carried out. Postoperative repeated AFP levels remained persistently elevated, in the range from 20 to 30 μg/mL. Careful evaluation for occult cancer showed no abnormality. Histology showed necrotic tissue and could not make a reliable diagnosis. A literature search was done using PubMed by key word “alpha-fetoprotein” and “elevation”. Thinking of a hereditary trait, we decided to screen patient's blood-related family members. Results AFP was found to be elevated in another four out of six relatives within three generations, unrelated to any disease. This pedigree was consistent with an autosomal dominant inheritance pattern. Conclusions HPAFP could mislead the physician. Failure to recognize HPAFP can lead to unsuitable treatments. The existence of this clinically benign condition needs to be considered in both children and adults with unexplained and persistent elevation of AFP, e.g. those diagnosed or suspected for germ cell tumor.

Kinetics of albumin- and alpha-fetoprotein-production during rat liver development

Synthesis of most of the plasma proteins is one of the main functions of the hepatocytes. Albumin synthesis is quantitatively the most abundant. In the present study we investigated albumin- and alpha-fetoprotein-gene-expression, and the function of the secretory apparatus during rat liver development. To this purpose we used the method of radioactive biosynthetic labeling of newly synthesized albumin and alpha-fetoprotein (AFP) to monitor the secretory capacity of endodermal cells derived from ventral foregut region (embryonic day 10, E10), and of embryonic and fetal hepatoblasts. Synthesis and secretion of albumin and AFP were already detected in the low numbered ventral foregut endodermal cells; fibrinogen synthesis was detectable in the E12 hepatoblasts, which were in higher number. The whole secretory machinery was functional from the earliest stages of liver development, and the speed of secretion was comparable with that of the adult hepatocytes. There was almost 4-fold increase of hepatoblasts cell volume in fetal stage compared with embryonic stage. The model used suggests that the hepatocyte secretory apparatus is already functional before the emergence of the liver bud. This is the first comparative report to analyze the hepatocyte secretory function, cell proliferation and cell volume during liver development.

Maternal alpha‐fetoprotein levels and sex of fetus

Dear Sir, Lei et al. 1 reported that women who gave birth to boys had higher alpha-fetoprotein (AFP) levels, on the average, than those who gave birth to girls. They gave evidence that this differential exists throughout pregnancy, and occurs even when birth weight is controlled. They cited other evidence for this differential, but expressed uncertainty concerning its cause. So, I would like to suggest a cause. Lei et al. 1 cite various lines of evidence to suggest that estrogen levels correlate with AFP levels, perhaps as a result of a direct causal process. They cite evidence, for example, that estradiol induces AFP synthesis in the mouse 2. A very large quantity of data has been adduced to support the hypothesis that the sexes of mammalian (including human) offspring are partially dependent on parental hormone levels around the time of conception 3, 4. Ex hypothesi, high maternal levels of estrogen around the time of conception are associated with the subsequent births of sons. The suggestion is that high levels of maternal estrogen first are causally responsible for the sons, and then, persisting through pregnancy, are responsible for the high AFP levels.

Expression and Localization of Alpha-fetoprotein mRNA and Protein in Human Early Villous Trophoblasts

Alpha-fetoprotein (AFP) is a major plasma protein produced during human fetal life. It is a good marker for several possible disorders affecting gestation. We previously reported that afp gene expression, which takes place mainly in yolk sac and fetal liver, also occurs in normal human placenta, specifically in early pregnancy. The aim of the present study was to determine the precise location of AFP synthesis sites within the placental villi. In situ hybridization and immunohistochemical experiments were performed on sections obtained from placentas of first-trimester and full-term pregnancies. We found that the pattern of afp gene expression was restricted to specific villous trophoblastic areas in early placentas. Both afp transcripts and AFP protein were mainly located in discontinuous regions, at junctions between two villi and at budding sites. In contrast, no AFP expression was detected in the cytotrophoblastic extravillous proliferative zone or in other placental cell types. According to the earlier studies, no AFP synthesis was detected in placental villous tissue from full-term pregnancies, using in situ hybridization and immunohistochemistry.

A two-dimensional electrophoresis preliminary approach to human hepatocarcinoma differentiation induced by PPAR-agonists

Adopting biochemical and proteomic approaches, we investigated the effect of some PPAR-agonists, a new class of differentiating agents, on human hepatocellular carcinoma Hep-G2 cell line. Cancer differentiation was assayed by checking albumin, transferrin and alpha-fetoprotein synthesis. Cell metabolism was studied by NMR spectroscopy of cell culture supernatants and by evaluation of mitochondrial respiratory chain enzyme activities. The two dimensional electrophoresis approach was employed to analyze modifications in the expression of cellular proteins linked to cell phenotype differentiation in the attempt to identify potential diagnostic and prognostic biomarkers of hepatocellular carcinoma. Results indicate that PPAR-agonists are able to act as differentiating inducers in human hepatocellular carcinoma Hep-G2 cell line as well as to inhibit mitochondrial respiratory chain Complex I, provoking a selective derangement of cellular oxidative metabolism. Lastly, two dimensional electrophoresis showed interesting modifications in the pattern of expression of cellular proteins that confirm biochemical data (increase in albumin and transferrin, decrease of alpha-fetoprotein synthesis) and, moreover, emphasize the meaning of these data by the increase of spots indicatively ascribed to HSP70 and catalase.

Elevated expression of bisectingN‐acetylglucosaminyltransferase‐III gene in a human fetal hepatocyte cell line by hepatitis B virus

UDP-N-acetylglucosamine: alpha-D-mannoside beta-1,4 N-acetylglucosaminyltransferase III (GnT-III) is a key enzyme in N-glycan biosynthesis. Human GnT-III enzyme activity was found to be elevated in the serum of patients with hepatomas and liver cirrhosis and in hepatocellular carcinoma tissues. Therefore, to understand the relationship between the elevation in GnT-III activity and hepatitis B viral (HBV) hepartocarcinogenesis, we investigated GnT-III gene expression in the HBV-infected cells. A cell line, HFH-T1, producing HBV was produced by natural infection of human fetal hepatocytes. A 170-bp band corresponding to the pre-S1 region of HBV was detected in the culture medium by polymerase chain reaction. Virions were also isolated from the culture medium by sucrose density gradient centrifugation. The synthesis of both alpha-fetoprotein and albumin as an indicator that these cells were functional hepatocytes and the extent of differentiation was examined. Polymerase chain reaction and Western blot analysis using a monoclonal antibody, GT273, which was prepared using human aglycosyl recombinant GnT-III were used for HBV DNA and GnT-III detection. Two types of HBV-related particles were secreted into the culture medium; one was a Dane particle (40 nm in size) containing HBV DNA and the other was a subviral hepatitis B surface antigen particle (20 nm in size) that did not contain the viral genome. The secretion from the cell line was diminished by the number of passages and, thus, this cell was renamed as HFH-T2. A decreased level of the HBV was secreted from the cells after a rest period. HFH-T2 cells showed a weak staining for alpha-fetoprotein and a moderate staining for albumin in the cytoplasm around the nucleus. High levels of a 0.7 kb DNA fragment originating from GnT-III DNA were detected in HFH-T2 cells. Western blot analysis using a monoclonal antibody, GT273, which was prepared using human aglycosyl recombinant GnT-III showed a single band, corresponding to Mr 63 kDa, whereas aglycosyl GnT-III showed a band at Mr 53 kDa, with a molecular weight difference of about 10 kDa. This indicates that HFH-T2 cells express glycosylated GnT-III. GnT-III activities were 347.2 +/- 53.6 pmol/mg of protein/h in HFH-T2, 276 +/- 26.3 in Hep3B, 252.5 +/- 23.3 in HepG2 and 30.7 +/- 3.4 in NIH-3T3. GnT-III activity was higher in HFH-T2 cells than in the hepatoma cell lines, Hep3B and HepG2. A human fetal hepatocyte cell line was transformed by infection with HBV and the cell line expressed high levels of GnT-III as the levels of secretion of HBV decreased. The decrease in HBV secretion from HFH-T2 cells could be due to a high level of expression of GnT-III. Such a cell line could be used to investigate relationships between HBV infection and glycosyltransferase gene expression. Furthermore, this cell line will be useful in future studies on the effect of the expression of GnT-III on other glycosyltransferase.

Alphafetoprotein and alphafetoprotein receptor expression in the normal human placenta at term

Alphafetoprotein (AFP) is detectable in maternal serum from around six weeks of gestation and is synthesised by the yolk sac and the fetal liver. The role of the placenta in the transport and possible synthesis of AFP is uncertain. The aim of this study was to investigate placental expression of AFP and the AFP receptor in uncomplicated pregnancies at term. Immunohistochemistry and Western blotting clearly demonstrated the presence of AFP in villous tissue at term. However, evidence of AFP mRNA expression or synthesis of AFP was not found following reverse transcription polymerase chain reaction of total RNA isolated from villous tissue and trophoblast cell cultures. The presence of a cell surface receptor for AFP in placental villous tissue, identified by immunohistochemistry and Western blotting, suggests a possible receptor-mediated mechanism for placental transport of AFP while the patterns of expression of AFP and its receptor may indicate a possible route by which AFP is transported across the placenta between the fetal and maternal circulations. These findings demonstrate that the placenta does not synthesise AFP at term and that the presence of AFP in the placenta is a reflection of transplacental transport of AFP possibly via a receptor-mediated mechanism.

Requirement of 3D extracellular network for maintenance of mature hepatocyte morphology and suppression of alpha-fetoprotein synthesis in vitro

It has been previously shown that adult mouse hepatocytes when co-cultured with rat liver-derived cell lines IAR-2 and IAR-20 can form organotypic hepatocyte islands consisting of cuboidal cells with expressed cell polarity and domain-specific localization of plasma-membrane proteins. The synthesis of alpha-fetoprotein, a fetal-specific antigen, was almost completely suppressed in these islands. It was noticed that organotypic islands were surrounded by fibrils and often covered with a roof of supporting cells. Here we demonstrate that the formation of adult mouse hepatocyte islands in co-cultures depends on the creation of 3D ECM scaffold formed by co-operative activity of hepatocytes and supporting partners. Noteworthy, the collagen type I network was observed exclusively around organotypic islands with complete suppression of alpha-fetoprotein synthesis. In contrast to non-transformed cells, IAR-2 cells, transformed either spontaneously or by transfection with mutant human N-ras Asn12 cDNA, were completely unable to support in co-culture the differentiated morphology of hepatocytes and to suppress AFP synthesis. Transformed IAR-2 cells were able to produce most of the main ECM components except collagen type I, but failed to form extracellular fibrils both in pure culture and in co-culture with hepatocytes. Together, the data show that 3D ECM scaffold (possibly based on collagen type I) plays an important role in maintaining the differentiated morphology of hepatocytes and suppression of alpha-fetoprotein synthesis in vitro.

Alpha-fetoprotein in human fetal cerebrospinal fluid

Alpha-fetoprotein (AFP) is a fetal glycoprotein. It has been ascribed a regulatory function of growth factor responses and immune functions. The concentrations of AFP and albumin (ALB) are highly variable in fetal serum and CSF and change with gestational age. The AFP index=[AFP CSF/AFP SERUM]/[ALB CSF/ALB SERUM] was determined in six normal fetuses at gestational age 17–23 weeks and found to be independent of gestational age and close to unity, mean 0.90±0.11 (S.D.). The ratio of CSF-serum concentrations of AFP and ALB both decreased significantly ( p<0.05) with gestational age. The mean fraction of AFP being non-reactive with concanavalin A was 1.7% in serum and 1.9% in CSF, suggesting a common hepatic origin of AFP in both compartments. In conclusion, the concentration of AFP in CSF seems to be determined largely by the serum-CSF concentration gradient in normal fetuses. This finding, combined with the remarkable constancy of the AFP index compared to the highly variable absolute concentrations of AFP in both serum and CSF should make the AFP index the marker of choice when analyzing for intrathecal AFP synthesis during development and in pathological conditions.

Influence of exogenous RAR alpha gene on MDR1 expression and P-glycoprotein function in human and rodent cell lines.

The goal of our study was to obtain direct evidence of co-ordinated regulation of P-glycoprotein (P-gp)-mediated multidrug resistance (MDR) and differentiation in tumour cells and to study some signalling pathways involved in joint regulation of these two cell phenotypes. The sublines of human melanoma (mS) and hepatoma (human HepG2 and rat McA RH 7777) cell lines were obtained by retroviral infection of the wild-type cells with the cDNA of the human retinoic acid receptor alpha (RAR alpha). The resulting sublines stably overexpressed exogenous RAR alpha gene. The infectants became more differentiated than the parental cells as determined by a decrease in the synthesis of the embryo-specific alpha-fetoprotein in HepG2 and McA RH 7777 hepatoma cells and by an increase in melanin synthesis in mS cells. The differentiation of human cells was accompanied by an increase in the amounts of MDR1 mRNA but not by an increase in P-gp activity as a drug transporter, in contrast, in the rat RAR alpha overexpressing cells P-gp functional activity was elevated. Treatment with cytotoxic drug (colchicine) or retinoic acid (RA) resulted in a slight increase in P-gp activity in the parental and RAR alpha-infected melanoma cells, whereas the increase in P-gp function in the infected hepatoma cells (both human and rat) was very prominent. Thus, we provide new evidence that cell differentiation caused by the overexpression of the gene participating in the differentiation programme leads to overexpression of MDR1 gene and drug resistance and that this effect is tissue and species specific. These data imply that the activation of the RA-controlled signalling pathway up-regulates MDR1 gene expression.

Rat yolk sac explants as a system for studying the regulation of endodermal genes: Down-regulation of the alpha-fetoprotein gene by clexamethasone and phorbol ester

The visceral yolk sac is a fetal membrane with essential placental functions. It is the major site of synthesis of alpha-fetoprotein (AFP), the most abundant plasma protein in the fetus. We developed a system of rat yolk sac explants in serum-free culture medium to study the regulation of endodermal gene expression in yolk sac. The explanted yolk sac tissues retained their double-sided morphology for up to 48 hours. The epithelial cells of both layers remained tightly joined on a basement membrane as seen by light and electron microscopy. This probably accounts for the continued expression of several endodermal cell-specific markers. The levels of mRNA encoding AFP, vitamin D-binding protein (DBP), hepatocyte nuclear factor 1alpha and beta transcription factors did not change during the 48-hour culture period. This reflects the stability of the differentiation state of the yolk sac endodermal cells. Dexamethasone and phorbol ester (TPA) specifically reduced the AFP mRNA level without affecting that of DBP. This suggests that these transduction pathways are functional in the yolk sac during this period of gestation and could be involved in the physiological down-regulation of AFP gene expression before birth. All these results show that this serum-free culture of rat yolk sac explants is a valuable system for further investigating the action of natural compounds and pharmacological drugs on endodermal gene expression during the embryonic and fetal periods.

Evaluation of the placenta: suggestions for a greater role in developmental toxicology.

Both in human and in rat, two types of placenta are present: the yolk sac (YS) and the chorioallantoic placenta. Histiotrophy, alpha-fetoprotein synthesis and blood cell formation occur in YS of both species. Besides, the midgut, primordial germ cells and possibly immunological structures originate from the YS tissue. The specialised cells of the chorioallantoic placenta attach the embryo to the uterus and form the vascular connections necessary for the nutrient transport. The placenta redirects maternal endocrine, immune and metabolic functions to conceptus advantage. These complex activities are sensitive to direct toxicity. Indirect effects on the placental functions might be elicited by immunomodulators and endocrine disrupters. Some experimental models could be utilised to identify possible toxic effects on placenta. Among the in vitro models the rodent giant yolk sac culture may be used to study the transport of materials, morphological and/or biochemical alterations and biotransformation activity of the visceral YS epithelium. Other in vitro approaches utilise human derived trophoblastic cells and tissues to investigate implantation and perimplantation toxicology. Besides specific studies, in vivo reproductive toxicity tests could pay more attention to the evaluation of placental tissues. Nowadays, some physiologically based pharmacokinetic models for developmental toxicity are also available to describe the disposition of toxic substances and their metabolites during pregnancy in rodents. Thus, more detailed studies on the embryo-foetal placenta may provide an important tool to understand developmental toxicity mechanisms, with particular regard to embryolethality and delayed development.

Regulation and activities of alpha-fetoprotein.

alpha-Fetoprotein (AFP) is one of the major serum proteins in the early life of mammals. The function of this protein is not yet fully understood. AFP is an oncodevelopmental gene product which is expressed at high levels in the embryonic yolk sac and fetal liver. The synthesis of AFP decreases dramatically after birth. Only trace amounts of AFP are synthesized in the adult liver. However, expression of the AFP gene is reactivated in the adult during liver regeneration and hepatocarcinogenesis. AFP is an excellent model system for studying the temporal and tissue-specific regulation of oncodevelopmental gene expression. Experiments with transgenic mice and DNA transfection studies have revealed several transcriptional control regions and cis-acting elements in the AFP gene. A large number of trans-acting protein factors interacting with these cis-acting elements have also been identified. Recent studies demonstrated that expression of AFP is regulated by at least two major signal transduction pathways in response to extracellular stimuli. The interactions between steroid hormone receptors and transcriptional factors which respond to separate signal transduction pathways result in transcriptional regulation of AFP gene expression. Trans-acting protein factors or steroid receptors complexed with given response elements can display different activities in different cell types due to cross-talk among both local protein-protein interactions within the DNA-binding domain, and distal protein-protein interactions. However, the detailed mechanisms of AFP gene expression are still not completely understood.

Serum levels of selected liver proteins following partial hepatectomy in the female rat.

Liver regeneration in the rat following partial hepatectomy (PH) is a frequently used model to study regulatory mechanisms in relation to cell growth and differentiation. In the present study, we analysed quantitative changes in the peripheral circulation of a number of important serum proteins following PH and laparotomy. Alpha-fetoprotein synthesis was induced in the PH rats and remained at very low levels in non-operated controls. Pregnancy-associated murine protein-1 levels and fibronectin levels were lower in the circulation of all operated animals compared with the intact controls. The serum levels of alpha-2 macroglobulin were elevated in all operated animals compared with the non-operated controls. Circulating complement factors C3c and C4 were present at significantly lower levels in PH animals than in rats following laparotomy. Small growth stimulating molecules (< 10 kDa) synthesized by different organs during the regenerative process have been described in the literature, but dialysates of the spleen and liver from non-operated rats and PH rats showed no significant impact on serum levels of the proteins in the present study.