Recombinant Alpha Research Articles

Preoperative Correction of Anemia to Allow a 3000 ml Blood Loss Without Transfusion in a Jehovah’s Witness Presenting for Explantation of an Infected Hip Joint Prosthesis: A Case Report

A 70-year-old Jehovah's Witness was treated with iron carboxymaltose intravenously, recombinant human erythropoietin alpha subcutaneously, and vitamin B12 and folate orally for 9 weeks to raise hemoglobin (Hb) from 10.8 to 17.0 g/dL before explantation of an infected hip joint prosthesis. The target Hb was calculated from the following formula: Hbtarget = Hbfinal/(1 - ABL/EBV), where Hbtarget= Hb to achieve before surgery, Hbfinal = lowest Hb patient could tolerate taking into consideration his comorbidities (7 g/dL), ABL = volume of blood the surgeon estimated the patient would lose intra- and postoperatively (3000 mL), and EBV = estimated blood volume (75 mL/kg for an adult man). Spinal anesthesia was provided with a single shot hyperbaric bupivacaine and fentanyl. Acute hypervolemic hemodilution was achieved with lactated Ringer's solution and hydroxyethyl starch. To further minimize blood loss, controlled hypotension to a mean blood pressure of 55 mm Hg was achieved with a propofol infusion and tranexamic acid was administered. Surgical blood loss was estimated to be 2500 mL. Hb at the end of surgery was 13.3 g/dL; on postoperative day 5, 11.7 g/L. No blood products were utilized.

Aspects of a Methodological Approach to Determination of Interferon Alpha Specific Activity

Specific antiviral activity is one of the key indicators characterising pharmaceutical quality and pharmacological efficacy of interferon alpha products (IFN-α). Specific activity is determined using a bioassay measuring antiviral activity in cell culture. The aim of the study was to select the most appropriate conditions for in vitro determination of IFN-α product specific activity. Materials and methods: Vero, MDBK, Hep-2, and A-549 homologous and heterologous cell cultures, as well as vesicular stomatitis Indiana virus (VSV) and murine encephalomyocarditis (EMC) virus at a dose of 100 TCD50 /0.1 mL were used for determination of specific antiviral activity. The international reference standard of recombinant interferon alpha-2b activity (Interferon alpha 2b, human, rDNA, E. coli-derived, 2nd WHO International Standard, 1999, NIBSC Code No. 95/566) and human recombinant interferon alpha 2b in the form of solution (batch No. 040214, Pharmapark LLC, Russia) were used as IFN-α samples. Results: the analysis of the obtained data helped to determine: the combinations of cell lines and the indicator virus most sensitive to IFN-α; the optimal concentration of fetal serum in the medium, and the optimal time parameters; the preferred method of reporting test results. Conclusions: the following test conditions were found to be optimal: the MDBK/VSV and Hep-2/EMC combinations proved to be the most sensitive to IFN-α; the optimal period of interferon and cell culture incubation—24 hours; the optimal concentration of fetal bovine serum in the culture medium used for diluting interferon products—2–5%. The instrumental procedure is preferred for reporting the results of interferon antiviral activity determination, because it is up-to-date, reliable, accurate and time-efficient.

A prospective multicenter randomized controlled study on the efficacy and safety of pharyngeal spraying recombinant human interferon alpha 2b in the treatment of children with herpangina

Objective To evaluate the efficacy and safety of pharyngeal spraying recombinant human interfe-ron alpha 2b (rhIFNα2b) in the treatment of herpangina in children. Method s A prospective, multicenter, rando-mized, opened and controlled study was carried out in 11 hospitals in Anhui province from August 2018 to March 2019.According to the time of admission, 180 patients diagnosed as herpangina were prospectively and randomly divided into rhIFNα2b treatment group and Ribavirin control group.On the basis of giving both groups the heat-clearing, detoxifying and anti-infection treatment, the patients in treatment group received pharyngeal spraying rhIFNα2b 9 g/L saline solution[1 million IU/mL, 0.1 million IU/(0.1 mL·press)], and the patients in control group were treated by pharyngeal spraying Ribavirin (0.5 mg RBV/press, 150 press), 3 presses per time, 4 times per day, continuous administration for 5 days for both groups.Those who recovered in advance were no longer given medication.All patients were observed to fully recover.The clinical efficacy and the disappearing time of symptoms and signs between two groups were compared, and the safety of pharyngeal spraying rhIFNα2b for patients was evaluated. Result s All of the 180 patients completed the study, including 90 cases in the treatment group and 90 cases in the control group.There was no statistically significant difference in terms of gender, age, weight and course of illness before treatment between the two groups (all P>0.05), which had clinical comparability.The apparent efficiency of the treatment group [63.3% (57/90 cases)] was significantly higher than that in the control group [38.9% (35/90 cases)] and the difference was statistically significant(χ2=10.934, P=0.004); no significant difference in the total efficiency between the treatment group [96.7% (87/90 cases)]and the control group [92.2% (83/90 cases)]was observed (χ2=2.924, P=0.169). The duration of fever[(32.59±20.73) h vs.(45.72±26.96) h], hyperemia[(76.48±23.12) h vs.(92.44±24.31) h], herpes[(72.99±25.77) h vs.(85.09± 26.62) h], salivation[(45.44±24.96) h vs.(54.42±31.20) h] and anorexia[(62.70±23.99) h vs.(78.71±30.54) h] in the treatment group were significantly shorter than those in the control group, and the differences were statistically significant(all P 0.05). After treatment, the serum levels of TNF-α and IL-6 were(7.26±1.99) ng/L and (2.42±0.73) ng/L in the treatment group, which were significantly lower than those in the control group [(12.09±6.39) ng/L and (7.32±11.51) ng/L](all P 0.05). The comparison on positive rate of virus in pharyngeal swab between the treatment group [65.3% (32/49 cases) and 40.6% (13/32 cases) respectively] and the control group[66.7%(36/54 cases) and 41.0% (16/39 cases), respectively]before and after therapy showed no significant difference (all P>0.05). During the treatment, no serious adverse reactions were observed in the two groups.The incidence of adverse reactions was 1.1% (1/90 cases) in the treatment group and 5.6% (5/90 cases) in the control group.In addition, the serum hemoglobin level of children in the control group after treatment was significantly lower than that before treatment and that in the treatment group (all P<0.05). Conclusions Compared with pharyngeal spraying ribavirin, pharyngeal spraying rhIFNα2b can greatly improve the clinical efficiency, accelerate the disappearance of clinical symptoms and signs, and shorten the total course of disease, and is more safe and worthy of clinical application. Key words: Herpangina; Recombinant human interferon alpha 2b; Ribavirin; Pharyngeal spraying

Effect of the Recombinant Adenovirus-Mediated HIF-1 Alpha on the Expression of VEGF in the Hypoxic Brain Microvascular Endothelial Cells of Rats.

ObjectiveTo investigate the effect of recombinant adenovirus-mediated HIF-1 alpha (HIF-1α) on the expression of vascular endothelial growth factor (VEGFA) and HIF-1α in hypoxic brain microvascular endothelial cells (BMEC) in rats.MethodsPrimary cultured rat BMEC in vitro were treated without or with either recombinant adenovirus-mediated hypoxia-inducible factor-1 alpha (AdHIF-1α) or recombinant adenovirus empty vector (Ad) in the presence of CoCl2 (simulating hypoxia conditions), or were grown under normoxia conditions. The expression of VEGFA and HIF-1α was analyzed at 12h, 24h, 48h and 72h incubation time, respectively. We also accessed a GEO dataset of stroke to analyze in vivo the alteration of HIF-1α and VEGFA expression, and the correlations between HIF-1α, VEGFA and CD31 mRNA levels in vascular vessels after stroke.ResultsVEGFA and HIF-1α expression were significantly higher in at each time point in the AdHIF-1α than other groups (p<0.05), whereas the Ad group and hypoxia group, showed no statistically significant difference (p>0.05). Moreover, VEGFA and HIF-1α levels were significantly higher in BMEC under hypoxia conditions than normoxia conditions (p <0.05). Both HIF-1α and VEGFA expression significantly increased after stroke in vivo with 1.30 and 1.57 fold-change in log2, respectively. There were significantly positive associations between HIF-1α, VEGFA and CD31 mRNA levels in vivo after stroke.ConclusionHypoxia-induced HIF-1α and VEGFA expression in vascular vessels, and recombinant AdHIF-1α could up-regulate VEGFA, and enhance HIF-1ααlevels in BMEC in vitro, which may play an important role in the recovery of stroke.

Immunization of rabbits with recombinant Clostridium perfringens alpha toxins CPA-C and CTB-CPA-C in a bicistronic design expression system confers strong protection against challenge

The Clostridium perfringens alpha toxin (CPA), encoded by the plc gene, is the causative pathogen of gas gangrene, which is a lethal infection. In this study, we used an E. coli system for the efficient production of recombinant proteins and developed a bicistronic design (BCD) expression construct consisting of two copies of the C-terminal (247–370) domain of the alpha toxin (CPA-C) in the first cistron, followed by Cholera Toxin B (CTB) linked with another two copies of CPA-C in the second cistron that is controlled by a single promoter. Rabbits were immunized twice with purified proteins (rCPA-C rCTB-CPA-C) produced in the BCD expression system, with an inactivated recombinant E. coli vaccine (RE), C. perfringens formaldehyde-inactivated alpha toxoid (FA-CPA) and C. perfringensl-lysine/formaldehyde alpha toxoid (LF-CPA) vaccines. Following the second vaccination, 0.1 mL of pooled sera of the RE-vaccinated rabbits could neutralize 12× mouse LD100 (100% lethal dose) of CPA, while that of the rCPA-C rCTB-CPA-C-vaccinated rabbits could neutralize 6× mouse LD100 of CPA. Antibody titers against CPA were also assessed by ELISA, reaching titers as high as 1:2048000 in the RE group; this was significantly higher compared to the C. perfringens alpha toxoid vaccinated groups (FA-CPA and LF-CPA). Rabbits from all vaccinated groups were completely protected from a 2× rabbit LD100 of CPA challenge. These results demonstrate that the recombinant proteins are able to induce a strong immune responses, indicating that they may be potentially utilized as targets for novel vaccines specifically against the C. perfringens alpha toxin.

Designing a light-activated recombinant alpha hemolysin for colorectal cancer targeting.

Introduction: Colorectal cancer (CRC) is one of the main health burden worldwide, which can cause major economic and physiological problems along with relatively high rate of mortality. It is important to develop new methods for the localized delivery of recombinant protein therapeutics, in large part due to the failure of conventional therapies in most cases. Since E. coli Nissle 1917 (EcN) does not produce any virulence factors, here we used these bacteria with the light-activated promoter system to deliver therapeutic agents in the desired location and time. Methods: In this study, Staphylococcus aureus alpha hemolysin (SAH), after codon usage optimization, was cloned into blue light activating vector (pDawn) and transferred to EcN strain. Then, the functionality and cytotoxicity of secreted alpha hemolysin was evaluated in the SW480 colon cancer cell line by using different experiments, including blood agar test, flow cytometry analysis, and DAPI staining. Results: Our findings revealed that EcN can produce functional SAH under the blue light irradiation against SW480 cancer cells. Moreover, cytotoxicity assays confirmed the dose- and time-dependent toxicity of this payload (SAH) against SW480 cancer cells. Conclusion: Based on our results, EcN is proposed as an appropriate light-activated vehicle for delivery of anticancer agents to the target cancer cells/tissues.

Recombinant Porcine Interferon Alpha Enhances Immune Responses to Killed Porcine Reproductive and Respiratory Syndrome Virus Vaccine in Pigs.

In this study, the immunoadjuvant effects of recombinant porcine interferon alpha (rPoIFNα) on the killed virus vaccine (KV) of porcine reproductive and respiratory syndrome virus (PRRSV) in pigs were investigated. The experimental pigs were divided into six groups, including normal control group, rPoIFNα control group, PRRSV KV control group, KV+40,000 U rPoIFNα immunization group, KV+400,000 U rPoIFNα immunization group, and KV+4,000,000 U rPoIFNα immunization group. The experimental pigs were boosted immunized on the 28th day after the initial immunization, and the heparinized blood and serum samples were collected at different time points of these two immunizations to detect and evaluate the immune responses of pigs after immunization by ELISA assay, neutralization assay, flow cytometry, and so on. The results showed that the proportion of the levels of PRRSV-specific antibodies, neutralizing antibodies, stimulation index, IL-4, IFN-γ, and lymphocytes within the groups immunized with KV+rPoIFNα were significantly higher than that group immunized with KV alone. The humoral and cellular immune responses in pigs were markedly enhanced by rPoIFNα after the coadministration with KV vaccine. Therefore, we tentatively think that rPoIFNα is a potential immune promoter with prospects for future applications in the pig industry.

Safety of aerosol inhalation of recombinant human interferon alpha 2b injection(P.putida)and excipient hydroxyethyl starch 40 to healthy SD rats

Objective To investigate the effects of recombinant human interferon alpha 2b injection(P.putida)and hydroxyethyl starch inhalation on physiological indexes and tissues and organs of SD rats, such as lungs, trachea and bronchus, and thus to explore the safety of the drug and excipients for inhalation. Methods Rats were randomly divided into two groups(hydroxyethyl starch 40 group and recombinant human interferon alpha 2b group).5 million IU/ml recombinant human interferon alpha 2b injection or 6% hydroxyethyl starch 40 injection were given respectively, which were diluted with 0.9% sodium chloride injection in a radio of 1: 2.Both groups were given aerosol inhalation for 30 minutes once a day for 14 days.Within 14 days after administration, the behavior of rats and the degree of toxicity were continuously observed.During the drug administration and recovery period, the clinical signs of the rats in each group were observed daily, and the body weight and food consumption were measured once a week.At the end of drug administration and the end of recovery period, hematology test, blood biochemical test, gross anatomical observation, organ weight determination, and histopathological examination were performed. Results The mass median aerodynamic diameter(MMAD)of the drug mist was 3.56 μm, and the geometric standard deviation(GSD)was 1.84 μm, which met the particle size requirements of arrival and deposition in the lower respiratory tract via atomization inhalation.No abnormality in clinical signs, body weight and food consumption, hematology and blood biochemical indexes were found in hydroxyethyl starch 40 group and recombinant human interferon alpha 2b group.No abnormal histopathological changes were observed in oral mucosa, tongue, nasal cavity(paranasal sinus), larynx, trachea, main bronchial tube, lung, heart, liver, spleen, kidney, reproductive system and other organs or tissues examined. Conclusion Results suggest the drug particle size of recombinant human interferon alpha 2b injection(pseudomonas)can reach the lower respiratory tract and deposit in the lower respiratory tract.Continuous atomization inhalation for 14 days has a good safety, and the excipient hydroxyethyl starch 40 has a high safety. Key words: Recombinant human interferon alpha 2b; Injection; Aerosol inhalation; Hydroxyethyl starch 40; Safety

Systems of intraganglia and intraneural control over latent alpha-herpesvirus in sensory ganglia of peripheral nervous system

The article presents modern data on interferon‑dependent mechanisms of control of latent alpha herpesvirus in the sensory nerve ganglia of the peripheral nervous system. The data on intraganglionar control mechanisms presented by resident natural killers, natural killer T‑lymphocytes and cytotoxic T‑cells that are part of the satellite cells of the nerve node, and the system of intraneuronal defense contained in the centripetal fibers and presented by cascade system TLR‑3, which leads to the production of pre‑immune interferons‑alpha, ‑beta and ‑lambda with a visorathic action, thus counteracting the centripetal transneural spread of the virus from the periphery to the central nervous system. The joint functioning of the interferon‑dependent systems of intraganglionar and intraneuronal control of alpha herpes viruses ensures the maintenance of the latent state of the pathogen, and the disruption of their work is associated with the risk of reactivation of the virus from latency with the development of clinical symptoms. At the same time, lack of intraganglionar cellular immunity is associated with frequent herpetic eruptions on the skin and mucous membranes, while a deficiency of intraneuronal TLR‑3‑dependent immunity leads to the development of herpes virus lesions of the central nervous system — myelitis, encephalitis, and meningitis. It supports the well‑known clinical fact, that herpetic encephalitis often develops in patients without a history of frequent specific exanthema, and individuals with frequent herpetic eruptions on the skin and mucous membranes may never suffer from viral lesions of the brain and spinal cord. New knowledge about the mechanisms of retaining alpha herpesviruses in the latent state of the sensory ganglia can help in the selection of rational therapy for immunocompromised patients with episodes of herpesvirus neuroinfections with the inclusion of drugs of natural, recombinant and lymphoblastoid human alpha interferons and various inducers of interferonogenesis of chemical or natural origin.The article will be useful, first of all, to neurologists, neurosurgeons, infectious diseases specialists, clinical immunologists, pathologists, physiologists.

Recombinant human soluble thrombomodulin ameliorates acetaminophen‑induced liver toxicity in mice

Recombinant human soluble thrombomodulin alpha (rhTM) has been developed as an anticoagulant with anti-inflammatory activity. Notably, acetaminophen (APAP) -induced liver disease (AILI) is caused by direct metabolite-induced hepatotoxicity as well as hepatic hyper-coagulation. To evaluate the utility of anticoagulant for the treatment of AILI, rhTM was administered in a mouse AILI model and liver damage was analyzed. AILI was induced in 8-week-old mice by intraperitoneal injection of APAP. rhTM (20 mg/kg) or placebo was injected at the same time as APAP administration. Serum alanine aminotransferase, fibrin degradation products and high-mobility group box 1 levels were significantly decreased in the rhTM-treated group compared with the control group. Furthermore, rhTM reduced the necrotic area and fibrin deposition in liver sections. rhTM suppressed the mRNA expression of heme oxygenase-1, plasminogen activator inhibitor type-1, tissue factors, and inflammatory cytokines compared with the control group. rhTM did not change the hepatic GSH content at 2 h after APAP injection, but restored them at 4 h after the insult. rhTM ameliorated liver damage in mice with AILI, probably via the improvement in liver perfusion induced by it's anticoagulant acitivity, which can lead to the suppression of secondary liver damage.

Molecular biology of trophoblast interferons and studies of their effects in vivo

Southern blotting of bovine genomic DNA indicated the presence of at least 3 bovine tIFN genes. The full DNA sequence of one of these genes, thought to be expressed in trophoblast, has been determined, including 193 bp of 5' non-coding region. The inferred amino acid sequence of bovine tIFN is more similar to ovine tIFN (80%) than to bovine IFN-alpha II (70%). The 5' flanking sequence has some similarity with bovine IFN-alpha II, and may contain a viral response element. A recombinant bovine alpha I interferon (Ciba Geigy; brIFN), resembling tIFN, extended oestrous cycle length in sheep when administered by intrauterine infusion over the period, Days 12-15 after oestrus, when maternal recognition of pregnancy occurs. Intramuscular injection was only effective at the doses used if given over a longer period (i.e. Days 9-15). Our experiments indicate that both tIFN and brIFN inhibit luteolysis by preventing a rise in endometrial oxytocin receptor concentrations, and suggest that tIFN achieves this by extending the time for which progesterone suppresses oxytocin receptor development. Further studies are required to confirm this hypothesis and to elucidate the interaction of the effects of progesterone and tIFN in endometrial cells.

High level extracellular production of recombinant human interferon alpha 2b in glycoengineered Pichia pastoris: culture medium optimization, high cell density cultivation and biological characterization.

The present study was aimed at design of experiments (DoE)- and artificial intelligence-based culture medium optimization for high level extracellular production of a novel recombinant human interferon alpha 2b (huIFNα2b) in glycoengineered Pichia pastoris and its characterization. The artificial neural network-genetic algorithm model exhibited improved huIFNα2b production and better predictability compared to response surface methodology. The optimized medium exhibited a fivefold increase in huIFNα2b titre compared to the complex medium. A maximum titre of huIFNα2b (436mgl-1 ) was achieved using the optimized medium in the bioreactor. Real-time capacitance data from dielectric spectroscopy were utilized to model the growth kinetics with unstructured models. Biological characterization by antiproliferative assay proved that the purified recombinant huIFNα2b was biologically active, exhibiting growth inhibition on breast cancer cell line. Culture medium optimization resulted in enhanced production of huIFNα2b in glycoengineered P. pastoris at both shake flask and bioreactor level. The purified huIFNα2b was found to be N-glycosylated and biologically active. DoE-based medium optimization strategy significantly improved huIFNα2b production. The antiproliferative activity of huIFNα2b substantiates its potential scope for application in cancer therapy.

A robust feeding control strategy adjusted and optimized by a neural network for enhancing of alpha 1-antitrypsin production in Pichia pastoris

Exact monitoring of the key process variables, such as specific growth rate (μ), is essential for effective process control, and maintenance of the optimal conditions for the production of recombinant proteins. In this study, a novel and confident on-line μ-stat approach for setting up methanol feeding strategy is described that is based on the consumption of ammonium hydroxide as feedback control. For this purpose a new and robust control with a time-based stable control structure was used, which had the ability to reconstruct the controller. Moreover, the MLP3 neural network was applied to adjust and optimize the performance of the robust control system. To evaluate the cited strategy, the overproduction of human recombinant alpha 1-antitrypsin (A1AT) under the control of alcohol oxidase 1 promoter (pAOX1) was investigated. On-line control of the μ at the optimal amount (0.03 1/h) led to 120 g/L dry cell weight and 324 mg/L of A1AT concentration. In comparison with traditional protocols of methanol feeding, the obtained product concentration demonstrated a significant improvement. It is suggested that this new presented method has a remarkable potential to be used for the overexpression of different recombinant proteins in the host Pichia pastoris as well as for process development in other hosts.

Choroid-plexus derived extracellular vesicles in Alzheimer’s disease and Parkinson’s disease: spreading the word

Event Abstract Back to Event Choroid-plexus derived extracellular vesicles in Alzheimer’s disease and Parkinson’s disease: spreading the word Charysse Vandendriessche1, 2*, Sriram Balusu3, Arnout Bruggeman1, 2, 4, Caroline Van Cauwenberghe1, 2, Griet Van Imschoot1, 2, Elien Van Wonterghem1, 2 and Roosmarijn Vandenbroucke1, 2 1 VIB-UGent Center for Inflammation Research, Belgium 2 Department of Biomedical Molecular Biology, Ghent University, Belgium 3 VIB & KU Leuven Center for Brain & Disease Research, Belgium 4 Department of Neurology, Ghent University Hospital, Belgium Objectives: The blood-cerebrospinal fluid barrier is formed by choroid plexus epithelial (CPE) cells, a single layer of epithelial cells situated at the interface between blood and the cerebrospinal fluid (CSF-) containing ventricular cavities. We could show that the CPE cells secrete more extracellular vesicles (EVs) into the CSF in the context of systemic inflammation (Balusu et al., EMBO Mol Med, 2016). We are currently studying this process in the context of Alzheimer’s disease (AD) and Parkinson’s disease (PD). Methods: We mimic AD or PD by the intracerebroventricular (icv) injection of respectively amyloid beta oligomers (AβO) or recombinant human alpha synuclein sonicated pre-formed fibrils (αSyn sPFFs) in wild type mice. Quantification of the amount of vesicles in the CSF is done using Nanoparticle Tracking Analysis (NTA) and the importance of the CPE cells as the source of EVs is studied using immunostainings, transmission electron microscopy and primary CPE cultures. Results: We found that after the icv injection of AβO or αSyn sPFFs, significantly more EVs are present in the CSF. We could show that at least an important part of these EVs are CPE-derived. Moreover, our models are associated with brain inflammation and the occurrence of behavioural deficits. Interestingly, we have indications that the CPE-derived EVs might play a role in these events. Conclusion: Our data show that AβO and αSyn sPFFs induce EV secretion into the CSF. Further research is needed to determine what their exact role in the pathogenesis of respectively AD and PD might be. Keywords: Choroid plexus (CP), Extracellular vesicle (EV), Alzheimer's disease (AD), Parkinson's disease (PD), Blood-cerebrospinal fluid barrier (BCSFB) Conference: 13th National Congress of the Belgian Society for Neuroscience , Brussels, Belgium, 24 May - 24 May, 2019. Presentation Type: Poster presentation Topic: Cellular/Molecular Neuroscience Citation: Vandendriessche C, Balusu S, Bruggeman A, Van Cauwenberghe C, Van Imschoot G, Van Wonterghem E and Vandenbroucke R (2019). Choroid-plexus derived extracellular vesicles in Alzheimer’s disease and Parkinson’s disease: spreading the word. Front. Neurosci. Conference Abstract: 13th National Congress of the Belgian Society for Neuroscience . doi: 10.3389/conf.fnins.2019.96.00008 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 30 Apr 2019; Published Online: 27 Sep 2019. * Correspondence: Mx. Charysse Vandendriessche, VIB-UGent Center for Inflammation Research, Ghent, Belgium, charyssevdd@irc.vib-ugent.be Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Charysse Vandendriessche Sriram Balusu Arnout Bruggeman Caroline Van Cauwenberghe Griet Van Imschoot Elien Van Wonterghem Roosmarijn Vandenbroucke Google Charysse Vandendriessche Sriram Balusu Arnout Bruggeman Caroline Van Cauwenberghe Griet Van Imschoot Elien Van Wonterghem Roosmarijn Vandenbroucke Google Scholar Charysse Vandendriessche Sriram Balusu Arnout Bruggeman Caroline Van Cauwenberghe Griet Van Imschoot Elien Van Wonterghem Roosmarijn Vandenbroucke PubMed Charysse Vandendriessche Sriram Balusu Arnout Bruggeman Caroline Van Cauwenberghe Griet Van Imschoot Elien Van Wonterghem Roosmarijn Vandenbroucke Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Corrective Effect of Gentabiferon-S on Weaned Piglet Immune Status and Its Effectiveness in Prevention of Intestinal Infections

The objective of the survey was to study the corrective effect of the Gentabiferon-S preparation on the immune status of the weaned piglets and its effectiveness in prevention of intestinal infections. A positive effect of the preparation on the morphological and immunological variables expressing the increased levels was ascertained. The red blood cell count, the relative lymphocyte count, the relative and absolute T–lymphocyte counts, the relative and absolute B-cell counts, the complement system activity, the lysozyme level, the neutrophil phagocytic activity, the number of phagocytes, the phagocytic index, the spontaneous and stimulated NBT test, the neutrophil activation reserve and index, and the amount of total immunoglobulins increased by 10.8, 16.5, 16.8 and 14.0%, 21.0 and 20.0%, 23.5, 66.8, 15.9, 38.2, 16.2, 11.0 and 21.7%, 13.3 and 17.1%, and 9.7%, respectively. A high (100%) effectiveness of the preparation in prevention of the intestinal infections is ensured by the presence of the antibiotic with broad-spectrum antimicrobial activity and the porcine recombinant alpha and gamma interferon.

Optimizing a recombinant estrogen receptor binding assay for analysis of herbal extracts

Phytoestrogens may directly or indirectly affect the function of estrogen receptors. A method using human recombinant estrogen receptors alpha and beta (ERα and ERβ) was adapted for pure isoflavone compounds: glycitein, glycitin, biochanin A and formononetin; and extracts from chaste tree, Queen-Anne’s lace, Cyperus rhizome, dong quai, flaxseed, black cohosh, Siberian ginseng, saw palmetto, hops, licorice, red clover and alfalfa. Estrogen binding equivalents (EBE) for extracts binding with ERα were: black cohosh < Queen-Anne’s Lace < saw palmetto < hops concentrate < licorice < red clover < alfalfa. Extracts from Cyperus, flaxseed, Siberian ginseng, chaste tree and dong quai had no detectable binding to ERα. For ERβ the estrogen binding equivalents were: chaste tree berry < hops concentrate <Cyperus rhizome < black cohosh < Siberian ginseng < dong quai < Queen-Anne’s Lace seed < flaxseed < saw palmetto berry < licorice < red clover < alfalfa. The relative binding affinity (RBA) of isoflavone standards for ERα and ERβ was compared to that of estradiol. At ERα, the RBA for isoflavone standards were as follows: glycitein > glycitin > biochanin A > formononetin and at ERβ the RBA were: biochanin A > glycitein > glycitin > formononetin. The use of recombinant ERα and ERβ receptor binding assays offer a relatively inexpensive and rapid technique that can be used as a tool to screen and identify plants with receptor binding activity.

Long-Term Efficacy and Safety of Recombinant Interferon Alpha-2 Vs. Hydroxyurea in Polycythemia Vera: Preliminary Results from the Three-Year Analysis of the Daliah Trial - a Randomized Controlled Phase III Clinical Trial

BackgroundRecombinant Interferon Alpha-2a (r-IFNα) is a potent immunomodulating agent, which has been used off-label for the treatment of polycythemia vera (PV) for more than three decades and has been demonstrated to induce high rates of clinical, hematological and molecular responses. Only few studies have compared efficacy and safety of r-IFNα vs. hydroxyurea (HU), which is considered first line therapy for PV patients > 60 years in most parts of the world. However, recent studies have provided encouraging results for the treatment of PV with r-IFNα compared to HU irrespective of age (R. Hoffmann 2016; H. Gisslinger 2018).AimsTo examine the difference in efficacy and safety of low-dose r-IFNα in PV patients ≤ 60 or > 60 years of age compared to HU > 60 years of age.MethodsNinety newly diagnosed or previously phlebotomized PV patients only (WHO 2008) were enrolled in the DALIAH trial (NCT01387763). All patients provided written informed consent. Patients ≤ 60 years were randomized (I:I) to r-IFNα-2a (Pegasys®) or to r-IFNα-2b (PegIntron®) whereas patients > 60 years were randomized (I:I:I) to either r-IFNα-2a, r-IFNα-2b or to HU. The starting dose of r-IFNα-2a and r-IFNα-2b was 45 or 35 µg/week, respectively. The HU dose was 500 to 2000 mg/day. Patients randomized to r-IFNα who presented with major thrombosis or platelets > 1500 109/L received HU from inclusion and until normalization of the platelet count. Efficacy assessment consisted of the clinicohematological and the molecular response rates by intention to treat analysis (ITT) using the European Leukemia Net (ELN) 2009 criteria. JAK2V617F analysis was performed by qPCR. Groups were compared by Fisher's Exact Test.ResultsThree-year analysis was available in all but five patients (n=85) at time of abstract submission (Table 1). The analysis was performed after a median of 36 months (range: 33-39 months). The median treatment dose was 684 mg/day (IQR: 131 - 942) for HU, 51 μg/week (IQR: 30-90 μg/week) and 54 μg/week (IQR: 30-66 μg/week) for r-IFNα-2a age ≤ 60 and > 60, respectively, and 41 μg/week (IQR: 29-45 μg/week) and 36 μg/week (IQR: 31-37 μg/week) for r-IFNα-2b age ≤ 60 and > 60.The overall clinicohematological response rate (ORR) was 68% (13/19) for HU, 42% (14/33) for r-IFNα ≤ 60 years and 39% (13/33) for r-IFNα > 60 years. The partial clinicohematological response rate (PHR) and the complete clinicohematological response rate (CHR) was 53% (10/19) and 16% (3/19) for HU, 9% (3/33) and 33% (11/33) for r-IFNα ≤ 60 years and 9% (3/33) and 30% (10/33) for r-IFNα > 60 years. Neither the ORR, CHR nor the PHR was significantly different between the three groups. Maintenance of CHR from first occurrence to data analysis after 36 months was 11% (2/19) for HU, 21% (7/33) for r-IFNα ≤ 60 years and 18% (6/33) for r-IFNα > 60 years.Forty-seven JAK2V617F positive patients were available for molecular response analysis after 36 months of therapy. A partial molecular response (PMR) was detected in 21% (4/19) of HU treated patients and in 24% (7/29) of r-IFNα treated patients ≤ 60 years and 18% (6/33) of r-IFNα > 60 years. Notably, 7% (2/29) of the r-IFNα treated patients ≤ 60 years obtained a complete molecular response (CMR). The median JAK2V617F reduction from baseline was 38% (IQR: 31-63%) for HU, 79% (IQR: 59-92%) for r-IFNα ≤ 60 years and 73% (IQR: 49-97%) for r-IFNα > 60 years. There was no statistically significant difference in the PMR between groups.Discontinuation of treatment for any reason after 36 months of therapy was 21% (4/19) for HU, 52% (17/33) for r-IFNα ≤ 60 years and 45% (15/33) for r-IFNα > 60 years. Toxicity related discontinuation was 5% (1/19) for HU and 30% (10/33) for both r-IFNα ≤ 60 and > 60 years. Grade 3-4 AEs occurred in 32% (6/19) of HU treated patients, 27% (9/33) in r-IFNα treated patients ≤ 60 years and in 42% (14/33) r-IFNα treated patients > 60 years. SAEs were reported in 21% (4/19) for HU, 9% (3/33) for r-IFNα ≤ 60 years and 24% (8/33) for r-IFNα > 60 years. The numbers of grade 3-4 AEs as well as SAEs were comparable between groups.ConclusionAfter 36 months of therapy CHR was non-significantly higher in PV patients treated with r-IFNα compared to HU by ITT, irrespective of age. Also, maintenance of CHR was longer for r-IFNα. However, ORR was non-significantly higher for HU. PMR was almost similar between the three groups but the median JAK2V617F reduction was greater for r-IFNα. Toxicity related discontinuation from study therapy was higher for r-IFNα compared to HU. DisclosuresStentoft:Bristol-Myers Squibb: Research Funding; Merck Sharp&Dohme: Research Funding. Hasselbalch:Novartis: Research Funding.

RETRACTED: Peptide-induced formation of protein aggregates and amyloid fibrils in human and guinea pig αA-crystallins under physiological conditions of temperature and pH

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the authors. The senior author contacted the journal in a forthright manner, in an effort to preserve the scientific integrity of the literature, after discovering a significant error in the results reported in the article. The authors were recently made aware of a paper by Kim et al. (Nature Commun. 2019) which shows a spirosome structure (the enzyme aldehyde-alcohol dehydrogenase) present in E. coli (Fig. 5a) that is very similar to the structure the authors thought formed when synthetic alpha A crystallin (66-80) peptide was incubated for 24 h with recombinant guinea pig alpha A insert crystallin (see Kumarasamy et al., Figs. 7C and F, and Fig. 9). Subsequent to publication of their report, the authors later found a number of images that showed what appeared to be the same structure present in samples of their presumably purified recombinant guinea pig alpha A insert crystallin which had been incubated without peptide for 24 h. Hence, the authors now conclude that the structures shown in Figs. 7C and F, and Fig. 9 of their article published in this journal are actually due to E. coli contaminant aldehyde-alcohol dehydrogenase. The authors deeply regret this error and any inconvenience it may have caused.

Design, Synthesis, and Pharmacological Characterization of Carbazole Based Dopamine Agonists as Potential Symptomatic and Neuroprotective Therapeutic Agents for Parkinson's Disease.

We have developed a series of carbazole-derived compounds based on our hybrid D2/D3 agonist template to design multifunctional compounds for the symptomatic and disease-modifying treatment of Parkinson's disease (PD). The lead molecules (-)-11b (D-636), (-)-15a (D-653), and (-)-15c (D-656) exhibited high affinity for both D2 and D3 receptors and in GTPγS functional assay, the compounds showed potent agonist activity at both D2 and D3 receptors (EC50 (GTPγS); D2 = 48.7 nM, D3 = 0.96 nM for 11b, D2 = 0.87 nM, D3 = 0.23 nM for 15a and D2 = 2.29 nM, D3 = 0.22 nM for 15c). In an animal model of PD, the test compounds exhibited potent in vivo activity in reversing hypolocomotion in reserpinized rats with a long duration of action compared to the reference drug ropinirole. In a cellular antioxidant assay, compounds (-)-11b, (-)-15a, and (-)-15c exhibited potent activity in reducing oxidative stress induced by neurotoxin 6-hydroxydopamine (6-OHDA). Also, in a cell-based PD neuroprotection model, these lead compounds significantly increased cell survival from toxicity of 6-OHDA, thereby producing a neuroprotective effect. Additionally, compounds (-)-11b and (-)-15a inhibited aggregation and reduced toxicity of recombinant alpha synuclein protein in a cell based in vitro assay. These observations suggest that the lead carbazole-based dopamine agonists may be promising multifunctional molecules for a viable symptomatic and disease-modifying therapy of PD and should be further investigated.

Attach great importance to innovative research and clinical application of recombinant human interferon alpha 1b in pediatrics

As the first class Ⅰ novel genetic engineering medical product in China, recombinant human interferon alpha 1b (IFNα1b) shows unique biological significance and clinical value.Especially in the field of pediatrics, IFNα1b has been used in the treatment of viral pneumonia, viral hepatitis, bronchiolitis, herpetic angina, hand-foot-mouth disease and some tumors in children and shown definite efficacy and good safety.It is also used in the prevention and treatment of virus-related wheezing and major infectious diseases, such as influenza.With a greater application prospect, IFNα1b plays an important role in promoting children′s health in China.Now, IFNα1b′s gene and molecular characteristics, new delivery methods, and the frontier clinical research will be discussed. Key words: Interferon α1b; Child; Virus; Inhalation; Wheezing; Influenza