Production Of Plasma Proteins Research Articles

In vivo liver targeted genome editing as therapeutic approach: progresses and challenges.

The liver is an essential organ of the body that performs several vital functions, including the metabolism of biomolecules, foreign substances, and toxins, and the production of plasma proteins, such as coagulation factors. There are hundreds of genetic disorders affecting liver functions and, for many of them, the only curative option is orthotopic liver transplantation, which nevertheless entails many risks and long-term complications. Some peculiar features of the liver, such as its large blood flow supply and the tolerogenic immune environment, make it an attractive target for in vivo gene therapy approaches. In recent years, several genome-editing tools mainly based on the clustered regularly interspaced short palindromic repeats associated protein 9 (CRISPR-Cas9) system have been successfully exploited in the context of liver-directed preclinical or clinical therapeutic applications. These include gene knock-out, knock-in, activation, interference, or base and prime editing approaches. Despite many achievements, important challenges still need to be addressed to broaden clinical applications, such as the optimization of the delivery methods, the improvement of the editing efficiency, and the risk of on-target or off-target unwanted effects and chromosomal rearrangements. In this review, we highlight the latest progress in the development of in vivo liver-targeted genome editing approaches for the treatment of genetic disorders. We describe the technological advancements that are currently under investigation, the challenges to overcome for clinical applicability, and the future perspectives of this technology.

Understanding the Potential Role of Nanotechnology in Liver Fibrosis: A Paradigm in Therapeutics.

The liver is a vital organ that plays a crucial role in the physiological operation of the human body. The liver controls the body's detoxification processes as well as the storage and breakdown of red blood cells, plasma protein and hormone production, and red blood cell destruction; therefore, it is vulnerable to their harmful effects, making it more prone to illness. The most frequent complications of chronic liver conditions include cirrhosis, fatty liver, liver fibrosis, hepatitis, and illnesses brought on by alcohol and drugs. Hepatic fibrosis involves the activation of hepatic stellate cells to cause persistent liver damage through the accumulation of cytosolic matrix proteins. The purpose of this review is to educate a concise discussion of the epidemiology of chronic liver disease, the pathogenesis and pathophysiology of liver fibrosis, the symptoms of liver fibrosis progression and regression, the clinical evaluation of liver fibrosis and the research into nanotechnology-based synthetic and herbal treatments for the liver fibrosis is summarized in this article. The herbal remedies summarized in this review article include epigallocathechin-3-gallate, silymarin, oxymatrine, curcumin, tetrandrine, glycyrrhetinic acid, salvianolic acid, plumbagin, Scutellaria baicalnsis Georgi, astragalosides, hawthorn extract, and andrographolides.

Stepwise access to safe plasma proteins in resource-constrained countries: Local production and pathways to fractionation-Report of an International Society of Blood Transfusion Workshop.

Actions are needed to improve access to safe plasma-derived medicinal products (PDMPs) in low- and middle-income countries (LMICs). The International Society of Blood Transfusion (ISBT) Working Party for Global Blood Safety organized an on-line workshop during 21-23 September 2021 to advance access to safe plasma proteins in resource-constrained countries, consistent with recent World Health Organization (WHO) guidance documents. The meeting drew attention to the considerable unmet needs for access to essential PDMPs in LMICs, in particular coagulation factors and immunoglobulins, and stepwise actions to address these deficits. First, improved access to safe plasma protein therapies requires blood component separation with prevention of wastage of recovered plasma. Quality and safety of collected blood and plasma must be assured so that plasma in excess of transfusion needs can be processed into safe plasma proteins. Second, local production of safe plasma proteins can be implemented using available technologies to locally obtain pathogen-reduced plasma and prepare pathogen-reduced cryoprecipitate and immunoglobulins from small plasma pools. Third, when a sufficient, stable volume of quality-assured plasma is available (approximately 50,000 L/year), contract or toll fractionation by a foreign plasma fractionator can expand the supply of PDMPs. Fourth, when the national infrastructure supports high-technology industrial production and stable volumes of quality plasma reach at least 200,000 L/year, technology transfer for domestic fractionation can be considered. Action is needed including commitments of the organizations that made the workshop possible (WHO, ISBT, World Federation of Haemophilia [WFH], Plasma Protein Therapeutics Association [PPTA], International Plasma Fractionation Association [IPFA], International Patient Organization of Primary Immunodeficiencies [IPOPI] and International Federation of Blood Donor Organizations [FIODS]).

Abstract 407: Belantamab Mafodotin detection by protein electrophoresis: Assessing interference for defining clinical response

Abstract Multiple Myeloma (MM) is a life threatening incurable cancer of the bone marrow resulting in plasma cell expansion and over production of plasma proteins or ‘M' proteins. Belantamab Mafodotin (Belamaf) is an antibody drug conjugate (ADC) for the treatment of multiple myeloma. The antibody component is an afucosylated IgG1 directed against B-cell maturation antigen (a protein expressed on normal B lymphocytes and multiple myeloma cells) conjugated to the microtubule inhibitor monomethylauristatin F (MMAF).Current criteria for assessing clinical response include changes in serum or urine M-protein levels by serum protein electrophoresis and immunofixation electrophoresis (IFE). Therapeutic use of ADC's has the potential to interfere with M-protein assessment by electrophoresis thus confounding clinical trial results. We aimed to modify an existing electrophoresis (EPS) platform (SAS-1 plus) to identify Belamaf in both saline and patients sera using a specific Anti-mcMMAF antibody.Objectives:1)To determine if Belamaf can be detected by serum protein electrophoresis and/or immunofixation electrophoresis methods that are used to measure ‘M' protein in MM patients. 2)If Belamaf can be detected by this assay, then determine if the detectable levels could interfere with defining clinical outcomes such as complete response*.By modifying an existing platform and using an mcMMAF specific antibody we were able to detect Belamaf in patients sera with no ‘M' protein at concentrations down to 0.5g/L by IFE. In multiple myeloma patients whose total protein concentrations were in excess of 90g/L Belamaf could be detected at 1.5g/L. Belamaf in patients sera can only be identified by anti-mcMMAF as anti-IgG and anti-IgKappa will detected those proteins present in sera in both MM and healthy patients.Belamaf could not be detected below 0.5g/L either by EPS or IFE using Anti-mcMMAF. As therapeutic Cmax levels of Belamaf are expected to be 0.04g/L, it will not be identified by EPS or IFE using the SAS-1 plus system. Further refinement of the SAS-1 plus system may achieve greater sensitive down to 0.2g/L (0.2g/L was detected using anti- IgG) however increased sensitivity and optimisation is highly unlikely to detected therapeutic concentrations of Belamaf.* Complete Response: Patients with CR must show that no abnormal monoclonal proteins (also called M-proteins) are present in the blood serum or urine. Citation Format: Fiona J. McClure. Belantamab Mafodotin detection by protein electrophoresis: Assessing interference for defining clinical response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 407.

Application of molecular farming based on barley plant (Hordeum vulgare L.) platform: A Review

Historically, humans have used plants as a source of medicine since ancient times. But the use of transgenic plants in molecular farming is a major source of molecular drugs. The use of molecular farming offers a promising prospect for producing of large-scale, low-cost pharmaceutical and industrial compounds. The results of this research include the production of plasma proteins, enzymes, growth factors, vaccines and antibodies. As plants can offer an inexpensive and convenient platform for the large-scale production of recombinant proteins with various functions, the driven force from the giant market for recombinant protein pharmaceuticals and industrial enzymes makes this subject grow and advance very quickly. With an increased demand for biopharmaceuticals, high costs, and inefficiency of current production systems, transgenic plants have received more attention as a new generation of bioreactors; but determining the best plant platform for the expression of recombinant proteins is one of the critical issues in molecular farming. So far, several expression platforms on this basis for the production of foreign proteins have been evaluated. Among these, the use of dry cereal seeds is one of the most efficient methods used to accumulate recombinant proteins. On the other hand, barley is a right candidate for recombinant protein expression platforms due to its easy cultivation management, high protein content, and low protein diversity, high adaptability to climatic conditions and low production costs. Besides, the highly self-pollinating nature of barley, along with its inability to produce fertile hybrids with related crops or wild species, creates a genetic mechanism, which practically prevents the unwanted dispersion of genetically modified germplasm under field conditions. Iran is also one of the centers of barley diversity in the Middle East, and the vast genetic variety, and nativeness of barley in this region is of particular importance to breeders.

Human liver model systems in a dish.

The human adult liver has a multi-cellular structure consisting of large lobes subdivided into lobules containing portal triads and hepatic cords lined by specialized blood vessels. Vital hepatic functions include filtering blood, metabolizing drugs, and production of bile and blood plasma proteins like albumin, among many other functions, which are generally dependent on the location or zone in which the hepatocyte resides in the liver. Due to the liver's intricate structure, there are many challenges to design differentiation protocols to generate more mature functional hepatocytes from human stem cells and maintain the long-term viability and functionality of primary hepatocytes. To this end, recent advancements in three-dimensional (3D) stem cell culture have accelerated the generation of a human miniature liver system, also known as liver organoids, with polarized epithelial cells, supportive cell types and extra-cellular matrix deposition by translating knowledge gained in studies of animal organogenesis and regeneration. To facilitate the efforts to study human development and disease using in vitro hepatic models, a thorough understanding of state-of-art protocols and underlying rationales is essential. Here, we review rapidly evolving 3D liver models, mainly focusing on organoid models differentiated from human cells.

Effects of Highly Active Antiretroviral Treatment on Liver and Kidney Functions

Aim: This study assesses the effects of HAART on liver and renal functions in HIV infected individuals on HAART.
 Study Design: Cross sectional study.
 Place and Methods: This study was conducted in Tamale, Ghana from August, 2015 to November 2017.
 Methodology: A total of 300 HIV infected participants with ages ranging from 19 to 79 years who have been administered with HAART for at least 6 months were recruited. Pre-HAART administration (baseline) demographic and clinical information, with initial liver and renal function test results were retrieved from the medical records of the participants present at the ART center. Post HAART administration blood sample (5 mLs) was taken from each participant into a gel separated vacutainer tube, allowed to clot and spun at 3000 rpm for 3 minutes to produce serum. The product (serum) was used for liver and renal function test analysis using a fully automated chemistry analyser (Vital Scientific Selectra Flexor XL).
 Results: Of the study population, 72% were administered with AZT/3TC/EFV, 13% with AZT/3TC/NVP, 6.7% with TDF/3TC/LPV/r and TDF/3TC/NVP, 1% with AZT/3TC/EFV while 0.7% were administered with TDF/FTC/EFV. The following parameters were significantly increased post HAART administration; ALT (25.53 ± 16.90 to 30.87 ± 19.28 U/L), ALP (163.7 ± 141.0 to 215.2 ± 143.4 U/L), GGT (37.27 ± 25.21 to 53.19 ± 41.71 U/L), Total protein (73.97 ± 17.08 to 82.31 ± 11.62 g/L), Albumin (38.02 ± 9.331 to 41.01 ± 7.471 g/L), Globulin 38.02 ± 15.71 to 42.79 ± 25.20 (g/L). There were however significant reductions in Total bilirubin (12.13 ± 10.85 to 9.434 ± 4.560 µmol/L), Direct bilirubin (6.616 ± 5.770 to 4.184 ± 2.806 µmol/L), (Creatinine 73.19 ± 36.13 to 63.14 ± 27.14 µmol/L) and Urea (3.515 ± 2.552 to 3.011±1.274 mmol/L).
 Conclusion: HAART improves renal function, induces elevation in liver enzymes, stimulates the production of plasma proteins and reduces serum bilirubin concentration.

Dietary management of liver disorders

The liver is responsible for many important functions, including metabolism of ingested nutrients and toxins, and production of plasma proteins, clotting factors and bile salts. When the liver is diseased or damaged, dietary therapy is beneficial in preventing further damage; supporting hepatic repair and liver function; and decreasing signs of hepatic encephalopathy. The nutritional status of patients with liver disease is often compromised by anorexia; nutrient malassimilation due to portal hypertension and cirrhosis; and decreased hepatic nutrient synthesis or storage capacity. Appetite and the amounts of protein, carbohydrates, fats, vitamins and minerals should all be considered. Previously, reduced dietary protein was often recommended; however, reducing dietary protein concentrations in all animals with liver disease is not appropriate. Nutraceuticals may have a beneficial effect in some cases. Nutritional support specifically for feline hepatic lipidosis, copper associated liver disease and hepatic encephalopathy are discussed.

The role of H19, a long non-coding RNA, in mouse liver postnatal maturation.

H19 RNA is highly expressed at early postnatal ages and precipitously decreases at a specific time corresponding with increases in expression of genes important for mature liver function, such as drug metabolizing enzymes. H19’s role in the regulation of liver maturation is currently unknown. Using an H19 knockout mouse model to determine the role of H19 in liver development, we quantified gene expression for insulin growth factor signaling, Wnt signaling, key cytochrome P450 (P450) enzymes known to change as the liver develops, and fetal and adult plasma protein produced in liver. In mice lacking H19 expression, liver weights were significantly increased immediately after birth and significant increases were found in the number of actively proliferating cells. Increases in cell proliferation may be due to increases in β-catenin protein affecting Wnt signaling, increases in insulin-like growth factor 2 (IGF2) expression, and/or increases in insulin-like growth factor 1 receptor (IGF1R) expression at the protein level. Loss of targeted inhibition of IGF1R by microRNA 675 (miR-675) may be the cause of IGF1R increases, as miR-675 expression is also abrogated with loss of H19 expression in our model. P450 expression patterns were largely unchanged. No change in the production of plasma proteins was found, indicating H19 may not be important for liver maturation despite its role in controlling cell proliferation during liver growth. H19 may be important for normal liver development, and understanding how the liver matures will assist in predicting drug efficacy and toxicity in pediatric populations.

New process for purifying high purity α1-antitrypsin from Cohn Fraction IV by chromatography: A promising method for the better utilization of plasma.

α1-antitrypsin (AAT) is a 52kDa serine protease inhibitor that is abundant in plasma. It is synthesized mainly by hepatic cells, and widely used to treat patients with emphysema due to congenital deficiency of AAT. A new isolation method for the purification of AAT from Cohn Fraction IV (Cohn F IV) is described. Cohn F IV is usually discarded as a byproduct from Cohn process. Using Cohn F IV as starting material does not interfere with the production of other plasma proteins and the cost of purification could be reduced greatly. Parameters of each step during purification were optimized, 15% polyethyleneglycol (PEG) concentration and pH 5.2 for PEG precipitation, elution with 0.05M sodium acetate and pH 4.7 for ion-exchange chromatography, and two steps blue sepharose affinity chromatography were chosen for AAT purification. The final protein with purity of 98.17%, specific activity of 3893.29 IU/mg, and yield of 28.35%, was achieved. Western blotting was applied for qualitative identification of final product, which specifically reacted with goat anti-human AAT antibody. LC-ESI-MS/MS was also employed to confirm the final protein. High performance liquid chromatography was used to analyze the composition of purified protein suggesting that pure protein was achieved. The molecular weight of AAT is 51062.77Da which was identified by LC-MS-MS. The manufacturing process described here may make better use of human plasma with Cohn F IV as starting material. The simple process described in this study is simple and inexpensive, it has a potential value for large scale production.

The genetic predisposition of complement C5 gene polymorphisms in proliferative diabetic retinopathy in Chongqing Han population

Objective To observe the genetic predisposition of complement C5 gene polymorphisms in proliferative diabetic retinopathy (PDR) in Chongqing Han population. Methods 400 type 2 diabetes (T2D) patients (case group) and 600 age- and sex-matched healthy controls (control group) were enrolled in this study. There were 8 PDR patients in case group. All the subjects were Han ethnic people. The immune-related representative SNP locus of C5 gene including rs2269067, rs7040033, rs7027797 were screened by linkage disequilibrium analysis. Locus rs1017119 was selected by TagSNP and was around the above three loci. Subjects' peripheral venous blood was collected and DNA was extracted. Genotyping was examined by PCR-restriction fragment length polymorphism method. The level of C5 plasma protein was measured by enzyme-linked immunoabsorbent assay. Results The frequency of GG genotype of rs2269067 was significantly increased in PDR patients in cases group compared with controls (Pc=3.4×10-5,OR=1.87, 95%CI=1.43 - 2.44; P=3.1×10-6). There was no differences in frequency of G, CC and CG genotype of rs2269067 between two groups (P=1.4×10-4, 1.000, 1.0×10-6). There were no differences in frequency of G, CC, CG, GG genotype of rs7040033, rs1017119, and rs7027797 between two groups (P>0.05). The production of C5 plasma protein was significantly increased in case group as compare with control group (P=0.0004). An increased production of C5 plasma protein was observed in rs2269067 GG genotype cases compared to CG or CC cases (P=0.003, 0.001). Conclusion C5 rs2269067 GG genotype may be associated with the PDR of T2D in Chongqing Han population. Key words: Diabetic retinopathy/etiology; Diabetic retinopathy/genetics; Complement C5/genetics; Genes/genetics; Polymorphism, single nucleotide/genetics

Book review: Production of Plasma Proteins for Therapeutic Use

Production of Plasma Proteins for Therapeutic Use edited by Joseph Bertolini, Neil Goss and John Curling provides comprehensive data for the plasma area. This comprehensive book covers the topics of the biology of plasma proteins and their production, from blood plasma collection to finished products, and also gives a description of these products' clinical use. Read this book review by Dr. Kjell Eriksson to find out more about this book.

Inflammatory Markers: Serum Amyloid A, Fibrinogen and C-Reactive Protein — A Revisited Study

The acute phase response is the part of the innate defence system of an animal against trauma, inflammation or infection. During this response, there is increased production and release of certain plasma proteins known as acute phase proteins, which include C-reactive protein (CRP), serum amyloid A (SAA) and fibrinogen (Fg). CRP consists of five identical subunits of 206 amino acids with a molecular weight of approximately 23 kDa. There is strong evidence from numerous studies that CRP is a predictor of inflammation. The acute-phase protein serum amyloid A (SAA) is a clinically useful marker of inflammation. SAA plays not only an important role in the development of AA amyloidosis (an important complication of rheumatoid arthritis) but also interacts with events closely involved in the metabolic syndrome as a high- and low-grade inflammatory modulator. Fibrinogen (Fg) is a high molecular weight plasma adhesion protein and is a biomarker of inflammation. It is synthesized and assembled in hepatocytes and fibroblasts and when secreted into the circulation, its plasma half-life ranges from 3 to 4 days. Several cytokines, are involved in the induction of acute phase protein synthesis, but the mutual importance of these cytokines seems to be cell-type specific and to vary in various experimental settings. Here we revisited the classic acute phase proteins SAA, C-Reactive protein and fibrinogen in their role in inflammation and their interrelationship with some cytokines.

Male Reproductive Traits and Their Relationship to Reproductive Traits in Their Female Progeny: A Systematic Review

The overall objective of one of the major research programs in the Co-operative Research Centre (CRC) for Beef Genetic Technologies is to 'Improve female reproductive performance' in tropical, northern Australian beef cattle herds. To address this overall objective, a quantitative genetics project focused on investigation of male reproductive traits was designed and linked to three female reproduction-focussed projects, (i) discovery of genes associated with post-partum re-conception and age at puberty; (ii) expression of genes associated with post-partum re-conception; and (iii) early predictors of lifetime female reproductive performance. During the initial planning of this male reproductive traits project, the CRC Scientific Review Committee recommended that the research team investigate and evaluate potentially new, early-life (i.e able to be measured before 2 years of age) predictors of both male and female reproductive performance. To address this recommendation, the following was carried out: (i) criteria for selection of traditional and candidate traits were established; (ii) methodology for tabulation of potential traits/phenotypes that define male and female reproductive function was developed; and (iii) a systematic scientific review of early-life predictors of male and female fertility was prepared. This review concluded that although factors that might be useful in predicting male reproductive performance have been studied for many years, there was relatively little useful information available to meet the objectives of this review. It was also concluded that the direction of future research should be guided not only by previous research which was scarce, but also by speculative hypotheses arising from an understanding of the physiological, endocrinological and genetic processes active in reproduction. A small number of new traits were recommended in addition to traditional sperm morphology, sexual behaviour, anatomical structure and growth traits. Potential additional traits include measurement of gonadotrophin-releasing hormone-stimulated luteinizing hormone (GnRH-stimulated LH); inhibin; several seminal plasma proteins (osteopontin, spermadhesin and seminal plasma proteins BSP30 and phospholipase A(2) could be used in an index); 11β-hydroxysteriod dehydrogenase; and leptin. In addition, the potential also exists to screen animals for a number of genetic markers associated with age of puberty, follicular recruitment and ovulation rate and genes associated with bovine seminal plasma protein and testosterone production. Insulin-like growth factor-1 (IGF-1) measurements are included because of their association with growth parameters, and an additional analysis demonstrated associations with male and female reproductive traits. Some of these factors have been previously evaluated in small numbers of animals of various species under intensive management conditions. Therefore, there is a need to evaluate these factors in much larger numbers of beef cattle grazing semi-extensive tropical production systems in northern Australia to determine their value in improving beef cattle enterprise profitability through improved herd fertility.

Recombinant plasma proteins

For almost 50 years, the fractionation of human plasma has been the sole possible source of a wide range of therapeutic proteins--such as coagulation factors, anticoagulants, immunoglobulins, and albumin--essential to the treatment of serious congenital or acquired bleeding or immunological diseases. In the last 20 years, the application of recombinant technologies to mammalian cell cultures has made possible--although with some limitations in productivity, costs, and immunogenic risks--to produce and commercialize complex plasma glycoproteins for human therapeutic applications and has opened the way to the development of new molecular entities. Today, the advanced exploration of alternative cell lines and enhanced cell culture systems, as well as the development of alternative expression technologies, such as transgenic animals, is opening a new era in the production of the full range of recombinant plasma protein therapeutics. In this review, we examine the achievements and ongoing challenges of the recombinant DNA technology as a platform for the production of plasma proteins and the advantages and limitations of such products compared to fractionated plasma proteins.

Hepatic AAV Gene Transfer and the Immune System: Friends or Foes?

Two ongoing clinical trials utilize different adeno-associated viral (AAV) vectors for liver-directed factor IX (F.IX) gene transfer with the goal of sustained therapy in patients with severe hemophilia B. Although preclinical studies have documented immune tolerance and long-term expression of F.IX in animals, the single prior clinical trial of this approach achieved only transient therapeutic gene expression and exposed preexisting immunity to the AAV vector as a major obstacle for therapy.1 Although accumulating preclinical data continue to fuel a debate over the potential impact of immune responses on hepatic AAV gene transfer, these data do not allow us to predict whether the human immune system will reject or tolerate therapy. This commentary dissects the relevant human and animal data, some of which are contradictory or allow us to draw only limited conclusions. More clinical trial data are critically needed and should ultimately help us develop better protocols.

HUPO’s Effort To Define the Liver Proteome

HUPO’s Effort To Define the Liver Proteome

Loss of Clotting Function in Mice Expressing a Mutant Form of Fibrinogen Lacking the α Chain Thrombin Cleavage Site

The conversion of soluble fibrinogen to an insoluble fibrin matrix following thrombin cleavage of the Aα and Bβ chains is critical to hemostasis. However, even soluble fibrinogen holds potential biological significance apart from fibrin polymer formation; for example, fibrinogen contributes significantly to overall blood rheology and can engage a number of integrins (e.g., αIIbβ3), enzymes (e.g., fXIII) and matrix proteins (e.g. fibronectin). To provide a system to study mice carrying fibrinogen but no capacity for fibrin formation, we employed a gene-targeting strategy to eliminate the Aα chain thrombin cleavage site and replace it with a sequence recognized by the highly-selective alternate protease, enterokinase (P6-P1: ADDDDK). Based on the prevailing view that thrombin cleavage of the Aα chain is essential to polymer formation, we hypothesized that this mutant fibrinogen, termed FibEK, would be locked in the soluble form in vivo, but would be readily clotted in vitro by exogenous enterokinase. The FibEK allele was transmitted through the germline and supported the expected level of mRNA expression and plasma protein production. Like previously established fibrinogen-null mice, FibEK mice uniformly developed to term. However, survival beyond the perinatal period was vastly superior in mice carrying fibrinogen-EK relative to fibrinogen-null animals in the same (C57Bl/6) genetic background; ~90% of FibEK mice survive to adulthood, whereas less than 30% of fibrinogen-null mice survive the perinatal period. Consistent with our initial hypothesis, FibEK plasma was found to be unclottable following the addition of excess exogenous murine thrombin at 37°C and complementary fibrinopeptide release assays by HPLC revealed that murine thrombin is completely incapable of releasing fibrinopeptide A (FpA) from fibrinogen-EK. Murine thrombin did support quantitative FpB release from fibrinogen-EK, albeit at a slower rate than wild-type fibrinogen. No thrombin-induced fibrin polymer formation could be appreciated by standard turbidity assays in incubation mixtures containing FibEK plasma at 37°C, even after 24 hrs. In contrast, wild-type plasma supported rapid polymer formation, with turbidity peaking within minutes of thrombin addition. Interestingly, despite the complete absence of FpA release, both plasma and purified fibrinogen-EK supported the formation of small and irregular clots (characterized by thin fibrils in scanning EM) following long incubations at reduced temperatures (i.e., 24 hr at 22 °C). Unlike fibrinogen-null mice, platelet-rich plasma prepared from FibEK mice supported robust ADP-induced platelet aggregation. However, FibEK animals essentially phenocopy fibrinogen-null mice in their inability to efficiently clear the microbial pathogen S. aureus. Following intraperitoneal infection with 109 CFU of S. aureus, control mice successfully cleared >99% of the bacteria within 1 hour, whereas the number of bacteria retrieved within peritoneal lavage fluid of both FibEK and FibAα−/− mice remained similar to the original input CFU. In summary, FibEK mice provide a unique opportunity to further explore the biochemistry of thrombin-mediated clot formation in vitro and in vivo. These animals will also be a vital tool in defining the specific contribution of fibrin matrices to broad array of physiological and pathological processes in vivo.

Pyrrolidine Dithiocarbamate Inhibits Interleukin-6 Signaling through Impaired STAT3 Activation and Association with Transcriptional Coactivators in Hepatocytes

Pyrrolidine Dithiocarbamate Inhibits Interleukin-6 Signaling through Impaired STAT3 Activation and Association with Transcriptional Coactivators in Hepatocytes

Liver cirrhosis and pregnancy

The liver accomplishes multiple functions such as the biotransformation of insoluble compounds (drugs, toxins, bilirubin), the metabolism and excretion of lipids (cholesterol in particular) the production of several plasma proteins (good examples are albumin, coagulation factors, transferrin), and the metabolism of amino acids and carbohydrates. The liver also plays a crucial role in the metabolism of different hormones, estrogens and progesterone among the others. This explains why during a chronic liver disease (CLD) severe alteration on the hormonal status occurs. Cirrhosis is often associated with amenorrhea and consequently, women with CLD (cirrhosis obviously included) have difficulty conceiving. If pregnancy occurs, it is associated with an increased risk of complications. Approximately 15 to 20 percent of pregnant women with CLD suffer spontaneous abortion, increased risk for premature childbirth or stillbirth. 1 If portal hypertension is present bleeding from esophageal varices is the biggest risk for women with cirrhosis. Variceal bleeding is most common during the second trimester, occurring in approximately 20 to 45 percent of women with portal hypertension. Variceal bleeding may also occur during labor. Death of the mother is similar to that occurring in the absence of pregnancy and accounts for about 20%. 2 It is recommendable that women with cir