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Background: There is a clear association between ABO blood group status and levels of Von Willebrand factor (vWF). Blood group O is related with lower expression of vWF which leads to relative bleeding tendency. Thus, relationships between Bleeding time (BT), Clotting time (CT) and blood groups are important in certain conditions like epistaxis, surgery, thrombosis etc. Aims and Objectives: i)To determine the blood group, of the students and to determine their relationship with BT and CT. Materials and Methods: Institution based cross-sectional observational study done in Department of Physiology, Tripura Medical College & Dr. BRAM Teaching Hospital. BT and CT were done by Duke’s Method and Capillary glass tube method respectively. Blood groups were determined on basis of presence or absence of agglutination. Qualitative data was expressed in percentage and quantitative data in frequency, mean and standard deviations. Chi-square test was applied. P value of 6 minutes found to be maximum in blood group O (53. 8%).Gender wise distribution showed CT > 6minutes in 39% females as compared to 34 %males (p value: 0.5826). Conclusion: In our study, CT was > 6 minutes and BT was > 4 minutes’ maximum in number in blood group O. Gender-wise BT and CT were higher in females than males. Our study showed that O blood group females are prone to certain diseases like epistaxis bleeding etc.

Polyvalent passive vaccines can act fast in resisting various bacteria with good efficacy, and they have application value in aquaculture. This study prepared live and inactivated Vibrio fluvialis mouse antisera (L-VF-antiserum and I-VF-antiserum), and administered them to goldfish (Carassius auratus) infected with V. fluvialis and Aeromonas hydrophila. The passive protective rates of live and inactivated mouse antisera against V. fluvialis were 60% (p < 0.05) and 40% (p < 0.05), and their passive cross-protective rates against A. hydrophila were 42.86% (p < 0.05) and 35.71% (p < 0.05), respectively. Furthermore, the two mouse antisera could recognize the bacteria in vitro; the content of bacteria in the C. auratus kidney decreased (p < 0.05), the phagocytic activity of C. auratus leukocytes was enhanced (p < 0.05), and the expression of inflammatory genes and activity of antioxidant factors decreased (p < 0.05). Moreover, the kidney, spleen, and intestinal tissue structures were intact, and the apoptosis and DNA damage were decreased (p < 0.05) among the kidney cells. The live V. fluvialis antiserum contained more antibodies against the outer membrane proteins of V. fluvialis than the inactivated mouse antiserum. The immunoprotective abilities of the live V. fluvialis antiserum were higher than those of the inactivated antiserum, and the antiserum of live V. fluvialis immunity demonstrated potential application value as a polyvalent passive immune vaccine in fish. [ABSTRACT FROM AUTHOR]

Mesoporous silica nanoparticles (MSN) are emerging as one of the most appealing candidates for gene carriers. Nevertheless, one major obstacle for their clinical application is the lack of efficient controlled release and gene delivery. In this study, a mesoporous silica‐modified gold nanorods (Au NRs) controlled release platform was synthesized. Then a thermosensitive copolymer was grafted to the surface of release platform, to impart the switching response of carrier under the photothermal of Au NRs. Fluorination were modified on the surface of MSN through a disulfide bond to achieve glutathione (GSH) response release and improve serum‐resistant ability. This gene carrier, with notable DNA condensation ability and negligible cytotoxicity, could be easily internalized into cells, enhanced green fluorescent protein expression in the presence of serum under laser irradiation. This work inspired us to solve the problem of traditional controllable sustained release and realize the performance of intracellular switch response to gene sustained release. [ABSTRACT FROM AUTHOR]

Copyright of Modern Food Science & Technology is the property of Editorial Office of Modern Food Science & Technology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Polyvalent passive vaccines can act fast in resisting various bacteria with good efficacy, and they have application value in aquaculture. This study prepared live and inactivated Vibrio fluvialis mouse antisera (L-VF-antiserum and I-VF-antiserum), and administered them to goldfish (Carassius auratus) infected with V. fluvialis and Aeromonas hydrophila. The passive protective rates of live and inactivated mouse antisera against V. fluvialis were 60% (p < 0.05) and 40% (p < 0.05), and their passive cross-protective rates against A. hydrophila were 42.86% (p < 0.05) and 35.71% (p < 0.05), respectively. Furthermore, the two mouse antisera could recognize the bacteria in vitro; the content of bacteria in the C. auratus kidney decreased (p < 0.05), the phagocytic activity of C. auratus leukocytes was enhanced (p < 0.05), and the expression of inflammatory genes and activity of antioxidant factors decreased (p < 0.05). Moreover, the kidney, spleen, and intestinal tissue structures were intact, and the apoptosis and DNA damage were decreased (p < 0.05) among the kidney cells. The live V. fluvialis antiserum contained more antibodies against the outer membrane proteins of V. fluvialis than the inactivated mouse antiserum. The immunoprotective abilities of the live V. fluvialis antiserum were higher than those of the inactivated antiserum, and the antiserum of live V. fluvialis immunity demonstrated potential application value as a polyvalent passive immune vaccine in fish.

The aim of the study was the serological and structural characterization of the lipopolysaccharide (LPS) O antigen from P. mirabilis Dm55 coming from the urine of a patient from Lodz. The Dm55 LPS was recognized in ELISA only by the O54 antiserum, suggesting a serological distinction of the Dm55 O antigen from all the 84 Proteus LPS serotypes described. The obtained polyclonal rabbit serum against P. mirabilis Dm55 reacted in ELISA and Western blotting with a few LPSs (including O54), but the reactions were weaker than those observed in the homologous system. The LPS of P. mirabilis Dm55 was subjected to mild acid hydrolysis, and the obtained high-molecular-mass O polysaccharide was chemically studied using sugar and methylation analyses, mass spectrometry, and 1H and 13C NMR spectroscopy, including 1H,1H NOESY, and 1H,13C HMBC experiments. The Dm55 O unit is a branched three-saccharide, and its linear fragment contains α-GalpNAc and β-Galp, whereas α-GlcpNAc occupies a terminal position. The Dm55 OPS shares a disaccharide epitope with the Proteus O54 antigen. Due to the structural differences of the studied O antigen from the other described Proteus O polysaccharides, we propose to classify the P. mirabilis Dm55 strain to a new Proteus O85 serogroup. [ABSTRACT FROM AUTHOR]

A medical narrative from a woman in her 90s describes her childhood bout with diphtheria in Kansas City, Missouri, apparently immediately after vaccination, her confinement in the "pest hospital," and her treatment with what she understood as a blood transfusion from a donor who was found through a radio appeal. In this essay, we trace the narrative back to the institutions, medical practices, and historical context, examining both the underlying history of medical practice and scientific understanding that is reflected in her experience and also the contexts of that history, including racial and religious attitudes. [ABSTRACT FROM AUTHOR]

Objective To prepare polyclonal antisera against enterovirus 71(EV71) capsid proteins. Methods Five B cell epitope prediction softwares were used to analyze capsid proteins of EV71 and four peptides against viral protein were synthesized, VP1(aa.204-223), VP2(aa.133-163), VP3(aa.139-148+aa.173-191) and VP4(aa.1-23). They were conjugated with keyhole limpet hemocyanin(KLH). BALB/c mice were subcutaneously immunized with six doses of KLH-peptides adjuvanted with MF59/CpGC274 at two-week interval. Serum sample was collected and examined with ELISA and Western blot. Results The four antisera were bound to both EV71 virus and denatured virus effectively. VP2 antiserum showed activity with denatured virus and the other three had similar binding activity to EV71 virus and denatured virus. The four antiserum samples were bound to both EV71 virus-like particle (VLP) and denatured VLP effectively. VP1 and VP2 antiserum could bind to VP1, VP0 of EV71 VLP separately. VP2 antiserum showed the highese binding activity and VP1 antiserum showed a inperior binding activity as compared to the formers. VP1 and VP2 antiserum were bound specifically and efficiently to EV71 virus. VP0 of empty particles and VP2 of intact EV71 virus particles could be simultaneously identified by VP2 antiserum. VP3 and VP4 antiserum failed to binding to no obvious binding to VLP and virus. Conclusions Four epitope peptide polyclonal antisera against EV71 are successfully prepared, which can provide ootential detection tools used in ELISA and Western blot and may be used for QC evaluation of EV71 vaccine.

Beet yellows virus, which belongs to the genus Closterovirus, family Closteroviridae and has a significant negative economic impact, has proven to be challenging to detect and diagnose. To obtain antibodies against BYV, we propose an easier bioinformatics approach than the isolation and purification of the wild virus as an antigen. We used the SWISS-MODEL Workspace (Biozentrum Basel) protein 3D prediction program to discover epitopes of major coat protein p22 lying on the surface of the BYV capsid. Sequences coding these epitopes were cloned into plasmid pQE-40 (Qiagen) in frame with mouse dihydrofolate reductase gene. Fused epitopes were expressed in Escherichia coli and isolated by the Ni-NTA affinity chromatography. Murine antibodies were raised against each epitope and in a combination of both and characterized by dot-ELISA and indirect ELISA. We successively used these antibodies for diagnosis of virus disease in systemically infected Tetragonia tetragonioides. We believe the approach described above can be used for diagnostics of difficult-to-obtain and hazardous-to-health viral infections. [ABSTRACT FROM AUTHOR]

The aim of the study was the serological and structural characterization of the lipopolysaccharide (LPS) O antigen from P. mirabilis Dm55 coming from the urine of a patient from Lodz. The Dm55 LPS was recognized in ELISA only by the O54 antiserum, suggesting a serological distinction of the Dm55 O antigen from all the 84 Proteus LPS serotypes described. The obtained polyclonal rabbit serum against P. mirabilis Dm55 reacted in ELISA and Western blotting with a few LPSs (including O54), but the reactions were weaker than those observed in the homologous system. The LPS of P. mirabilis Dm55 was subjected to mild acid hydrolysis, and the obtained high-molecular-mass O polysaccharide was chemically studied using sugar and methylation analyses, mass spectrometry, and 1H and 13C NMR spectroscopy, including 1H,1H NOESY, and 1H,13C HMBC experiments. The Dm55 O unit is a branched three-saccharide, and its linear fragment contains α-GalpNAc and β-Galp, whereas α-GlcpNAc occupies a terminal position. The Dm55 OPS shares a disaccharide epitope with the Proteus O54 antigen. Due to the structural differences of the studied O antigen from the other described Proteus O polysaccharides, we propose to classify the P. mirabilis Dm55 strain to a new Proteus O85 serogroup.

The technology of in vitro micropropagation of the apple rootstock MM 106 was developed. From 80 to100% rooting was obtained in variants with indole-3-butyric acid (IBA) 1-2 mg/l. A diagnostic polyclonal antiserum to the apple stem pitting virus (ASPV) was obtained. The results of the virus elimination of apple cultivars 'Coreor', 'Coredova', 'Coretard', 'Izvoras' genetically resistant to scab Venturia inaequalis Cooke Wint using methods of thermotherapy are described. The virus free clone’s category Prebase were planted in the depozitarium of the institute in order to increase and preserve the virus-free genefond of pome fruit crops.

Porcine circovirus (PCV) comprises four types, PCV1, PCV2, PCV3, and PCV4, which belong to the Circovirus genus of the family Circoviridae. PCV1 is nonpathogenic, whereas PCV2, PCV3, and PCV4 can infect pigs and cause disease. However, due to a lack of experimental evidence, whether vaccines based on PCV capsid (Cap) can induce cross-reactivity against PCVs remains controversial. In this study, recombinant truncated capsids (rCaps) of PCV2, PCV3, and PCV4 were highly and efficiently expressed and purified, followed by the development and evaluation of antibodies against PCVs. The results showed that monovalent and trivalent antigens based on the recombinant Caps had adequate immunogenicity to stimulate specific antibodies against the corresponding protein and virus. Furthermore, antisera prepared from the recombinant Caps also cross-reacted with different PCVs. Therefore, recombinant proteins can be used as candidate antigens to develop vaccines and ELISA diagnostic kits. In addition, the antibodies prepared in this study are promising candidates for the simultaneous prevention and treatment of PCVs in the clinic. [ABSTRACT FROM AUTHOR]

Rehmannia glutinosa is a top-grade traditional Chinese medicine, and also is an important planting medicinal material for Chinese poor farmers shaking off poverty. Rehmannia mosaic virus (ReMV) causes big economic loss of R. glutinosa in planting area. However, there is no effective methods for quick, accurate, and high-throughput detection for ReMV in Chinese production area. The preserved R. glutinosa samples carrying ReMV was taken for research material. The coat protein coding sequences (CPReMV) was cloned and sequenced. The target sequence was further placed into a prokaryotic expression vector to express the N-terminal-tagged recombinant CPReMV protein (His-CPReMV). Purified His-CPReMV was used as an antigen to immunize New Zealand white rabbits, and antiserum was obtained. The titers and sensitivities of the antisera were analyzed and evaluated. Polyclonal antibodies were purified from the antiserum, and the titers and sensitivity to the target His-CPReMV protein were evaluated. The results demonstrate that the obtained polyclonal antibodies against His-CPReMV could be successfully used for rapid, accurate, and high-throughput detection of ReMV from R. glutinosa planted in the wild. Our investigation established serological-based detection methods for ReMV for the first time, and provides a foundation for future exploration of the pathogenic mechanisms of ReMV in R. glutinosa.

Copyright of Journal of Dalian Polytechnic University is the property of Journal of Dalian Polytechnic University Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

α-conotoxin (α-CTX) MI is a small peptide toxin with 14 amino acids and two disulfide bonds. It potently inhibits muscle-type nicotinic acetylcholine receptors (nAChRs), and poses a threat as a toxin to tropical fishermen. However, there are currently no effective drugs for the treatment of MI envenomation due to the toxin's low immunogenicity. In this report, we generated neutralizing antiserum and F(ab') 2 to MI by synthesizing a new MI antigen through the coupling of alkynyl-modified MI and azide-modified bovine serum albumin (BSA), followed by immunization into mouse and horse. The new MI-BSA antigen generated high titers of mouse and horse antiserum (1:204,800 and 1:51,200, respectively), and both the antiserum as well as the horse F(ab') 2 displayed highly potent neutralization and detoxification efficacy. 12.5 μL of mouse or horse antiserum preincubated with MI could completely neutralize a lethal dose of the MI (0.4 μg, 1.7 × LD 50), while 6.25 μL (mouse) or 10.41 μL (horse) of the antiserum could exert complete detoxification of mice injected with 1.7 × LD 50 of MI. Moreover, the mouse and horse antiserum exhibited medium cross-reactivity for highly toxic α-CTX GI. These results demonstrate that the integrity of MI's antigen epitope and carrier effect of BSA can improve MI's immunogenicity, and provides an effective detoxification treatment for highly toxic α-conotoxins as well as an effective method for the preparation of antiserum of small peptide toxins. [Display omitted] • α-conotoxin MI-bovine serum albumin conjugate antigen was synthesized by click reaction. • This antigen maintains the integrity of MI's antigen epitope. • This antigen displays high immunogenicity. • This antigen produces antiserum with high neutralizing and detoxification activity. • The MI's antiserum possesses medium cross-reactivity to α-conotoxin GI. [ABSTRACT FROM AUTHOR]

Background: Scorpionism is a worldwide medical issue, especially relevant in the tropical and subtropical countries. Tityus serrulatus is the species responsible for most cases in Brazil. Antivenom administration to victims is the sole specific therapy obtained from donor animals. Most of these donors suffer with symptoms of the poisoning, debilitating their health and reducing their life expectancy. The aim of the present research was to evaluate whether the immunogens prepared from the crude and detoxified venom of T. serrulatus promoted different changes in fractionated sheep plasma proteins, during a scorpion antivenom serum production. Materials, Methods & Results: Twelve sheep, healthy, mean weight of 30 kg, were distributed into 3 groups (n = 4): G1 (control), G2 (crude venom) and G3 (detoxified venom). The adopted immunization protocol (first cycle) had 6 doses, 3 using Freund’s adjuvant, with a 21-day interval between each one (day 0, 22 and 43), and 3 doses with no adjuvant (booster) and 0.2 mg of antigen (reinforcement), spaced 3 days between each other (day 50, 53 and 56). Group control (G1) received 6 immunizations with phosphate buffered saline (PBS) associated with Freund’s adjuvant (1:1), while the other 2 groups received 0.5 mg of venom (G2) and detoxified venom (G3), respectively, diluted in PBS, associated with the Freund adjuvant. The boosters were 1/3 of the initial dose, diluted only PBS. At baseline (T0) and at 24 and 48 h after immunization, all animals underwent clinical examinations. Blood samples were collected at day 0, 22, 43, 53 and 56 for proteinogram analysis. Total protein, albumin and globulins fractions were measured. Plasma albumin concentration at T0 ranged from 3.41-4.86 g/dL, with a mean value of 4.12 g/dL. There was no statistical difference between the 3 experimental groups. The normal values determined for α-globulin range from 0.14 to 0.54 g/dL, with a mean of 0.31 g/dL (T0). There was a significant increase in the 3rd immunization and its respective interval (24-48 h), with values above normal in all groups: G1 (0.66 g/dL), G2 (0.62 g/dL) and G3 (0.65 g/dL). The β-globulin was subdivided into β1 and β2 globulin. At T0, the normal values of β1 ranged from 0.45 to 1.05 g/dL, with a mean of 0.664 g/dL, and no significant change was observed in this classification. On the other hand, there was an abrupt increase in β2 in all groups after the first immunization, compared to the baseline value in T0 (0.37 g/dL mean value). From the third to the 6th immunization, there was an important reduction in β2 fraction when compared with baseline value. The γ-globulins fraction ranged from 0.80 g/dL to 76 g/dL. In the 6th immunization, there was a significant difference between G1 and the groups that received venom (G2 and G3). Therefore, all animals presented an acute inflammatory response, evidenced by the significant reduction of plasma albumin and an increase in α-globulin and β2 -globulin. It is important to point out that T. serrulatus detoxified venom did not cause alterations in ovine proteinogram during the first cycle of immunization. Discussion: The fact that both groups (G2 and G3) presented acute inflammatory response, indicates that this alteration is caused by the adjuvant present in the immunization protocol. Tityus serrulatus venom detoxified with glutaraldehyde did not cause significant alterations in ovine proteinogram in the early stages, suggesting that it may be used as an alternative antigen for the production of antivenom, improving clinical conditions of donor animals. [ABSTRACT FROM AUTHOR]

Brucellosis, caused by Brucella species, is common among humans and animals, and is one of the most common infectious diseases in Iran. Several assays are available to detect brucellosis, but serological tests may be the only method used in many laboratories. In Iran, different kits are produced in the Pasteur Institute based on agglutination, such as Rose Bengal and 2-mercaptoethanol (2-ME). The positive antiserum control used in these kits is produced in rabbits. The purposeof this study was to produce the antiserum in goats and to compare the titer, quality, and quantity with the antiserum produced in rabbits. The goat immunization was performed by intramuscular injection. Seven days after the last injection, sera were collected. The produced antibody was used in slide and tube agglutination tests with different antigens. The results indicated that the antiserum, produced by the goats had a high quality and quantity. Slide agglutination test showed positive results at 1/6400 dilution with goat antiserum (4+) and 1/1600 dilution with rabbit antiserum (1+). The application of the goats is a better and more appropriate choice, in terms of both cost and quantity, when a high concentration of serum is required. In addition, one goat can provide a higher amount of antiserum compared to several rabbits. HIGHLIGHTS • This study presents a robust and time-saving method for the production of an efficient antiserum. • Goat is a better and more appropriate host when higher amount of serum is required. • The amount of serum produced in goat is approximately 8 times greater than that of rabbit.

The use of the serum or plasma of patients or animals who have recovered from an infectious disease, or had been immunized with a relevant antigen, to treat or prevent the same infection in others began in the late 1880s when French and German scientists uncovered, one step at a time, several of the elements of the immune system's response to infection. A key finding was that the damage caused by some bacteria depends upon their secreted toxins which can be neutralized by biologic agents. Antitoxins to diphtheria and tetanus began to be manufactured in large animals in France, Germany, and the US in the 1890s and were soon being used worldwide. The impact of diphtheria antitoxin on childhood mortality was profound. Shortly after the development of antitoxins, convalescent serum began to be used for its anti-bactericidal properties thus addressing serious infections caused by non-toxin-producing organisms. The effectiveness of antitoxins and antisera was demonstrated by examining mortality rates in hospitals before and after the introduction of antitoxins, by comparisons of treated and untreated patients, by comparing early and late treatment and dosage, by examining vital data mortality trends, and by several randomized and alternate assignment trials. Antitoxins continue to have a role in the rare cases of diphtheria and other conditions largely eradicated by immunization, but serum therapy nearly disappeared from the medical armamentarium with the development of antibiotics in the 1940s. Inasmuch as new human pathogens are now emerging with unprecedented regularity as seen in the recent COVID-19 pandemic, and because specific therapies are unlikely to be available for them, plasma-based antibody therapies are likely to again carve out a niche in infectious disease control., (© 2024. The Author(s), under exclusive license to Springer Nature Switzerland AG.)

Beet yellows virus, which belongs to the genus Closterovirus, family Closteroviridae and has a significant negative economic impact, has proven to be challenging to detect and diagnose. To obtain antibodies against BYV, we propose an easier bioinformatics approach than the isolation and purification of the wild virus as an antigen. We used the SWISS-MODEL Workspace (Biozentrum Basel) protein 3D prediction program to discover epitopes of major coat protein p22 lying on the surface of the BYV capsid. Sequences coding these epitopes were cloned into plasmid pQE-40 (Qiagen) in frame with mouse dihydrofolate reductase gene. Fused epitopes were expressed in Escherichia coli and isolated by the Ni-NTA affinity chromatography. Murine antibodies were raised against each epitope and in a combination of both and characterized by dot-ELISA and indirect ELISA. We successively used these antibodies for diagnosis of virus disease in systemically infected Tetragonia tetragonioides. We believe the approach described above can be used for diagnostics of difficult-to-obtain and hazardous-to-health viral infections.

Klebsiella spp. and Proteus spp. cause hospital-acquired urinary tract infections (UTIs), which are often related to the use of catheters. To create a vaccine preventing UTI, immunogenic bacterial antigens with common epitopes are still being looked for. In this work, the role of polysaccharide antigens of four Klebsiella spp. and eight Proteus spp. strains in serological cross-reactions with specific antisera was examined. Enzyme-linked immunosorbent assay (ELISA), Western blotting, and silver staining by Tsai method were performed. The Klebsiella and Proteus spp. LPSs and cells were used as antigens. Polyclonal rabbit sera specific to Klebsiella oxytoca 0.023 and 0.062 strains and four Klebsiella spp. LPSs were obtained. The ELISA and Western blotting results showed the strongest cross-reactions occurring between lipopolysaccharides (LPSs) from four Klebsiella strains and P. vulgaris O42 antiserum. The silver-staining procedure revealed the patterns typical of both slow- and fast-migrating mass species of the Klebsiella LPSs. The Klebsiella spp. antigens also cross-reacted with four P. penneri antisera, and most of the reactions were observed as low-migrating patterns. From two K. oxytoca antisera obtained in this work, only one, the K. oxytoca 0.062 antiserum, cross-reacted with satisfactory strength with P. penneri LPSs (19, 22, and 60). Obtaining cross-reactions between the antigens of Klebsiella strains and Proteus antisera and in the opposite systems is important for proving the immunogenic role of polysaccharide antigens in triggering the immunological response.

Klebsiella spp. and Proteus spp. cause hospital-acquired urinary tract infections (UTIs), which are often related to the use of catheters. To create a vaccine preventing UTI, immunogenic bacterial antigens with common epitopes are still being looked for. In this work, the role of polysaccharide antigens of four Klebsiella spp. and eight Proteus spp. strains in serological cross-reactions with specific antisera was examined. Enzyme-linked immunosorbent assay (ELISA), Western blotting, and silver staining by Tsai method were performed. The Klebsiella and Proteus spp. LPSs and cells were used as antigens. Polyclonal rabbit sera specific to Klebsiella oxytoca 0.023 and 0.062 strains and four Klebsiella spp. LPSs were obtained. The ELISA and Western blotting results showed the strongest cross-reactions occurring between lipopolysaccharides (LPSs) from four Klebsiella strains and P. vulgaris O42 antiserum. The silver-staining procedure revealed the patterns typical of both slow- and fast-migrating mass species of the Klebsiella LPSs. The Klebsiella spp. antigens also cross-reacted with four P. penneri antisera, and most of the reactions were observed as low-migrating patterns. From two K. oxytoca antisera obtained in this work, only one, the K. oxytoca 0.062 antiserum, cross-reacted with satisfactory strength with P. penneri LPSs (19, 22, and 60). Obtaining cross-reactions between the antigens of Klebsiella strains and Proteus antisera and in the opposite systems is important for proving the immunogenic role of polysaccharide antigens in triggering the immunological response. [ABSTRACT FROM AUTHOR]

T lymphocytes play an important role in cellular and adaptive immunity in vertebrates. The mechanisms of the fish immune system are little studied because of the lack of population‐specific antibodies. This study examined the expression of two T lymphocyte markers, TCRα (PoTCRα) and CD8α (PoCD8α) in the Japanese flounder (Paralichthys olivaceus). The expression of PoTCRα and PoCD8α was mainly detected in immune/mucosal tissues. Recombinant PoTCRα and PoCD8α were expressed in pET32a and pET259, respectively. Then, rabbit anti‐PoTCRα serum and rat anti‐PoCD8α serum were prepared. Using serum, the characteristics of TCR+ and CD8+ head kidney leucocytes (HKLs) were investigated. The results of laser scanning confocal microscopy (LSCM) demonstrated that TCRα and CD8α were transmembrane proteins localized on the cell surface. The populations of CD8α−, CD8α+, TCRα−, and TCRα+ were sorted by flow cytometry (FCM) and analysed using qRT‐PCR. The results demonstrated that all TCRα+/TCRα− or CD8α+/CD8α− HKLs expressed IFN‐γ. The CD4‐1 and IgM transcripts were detected only in TCRα− and CD8α− cells. Furthermore, HKL mitogenesis was induced with concanavalin A (ConA) stimulation. Taken together, the results from LSCM and FCM analyses showed that mammalian and P. olivaceus TCR+ and CD8+ leucocytes share basic characteristics. [ABSTRACT FROM AUTHOR]

Rotavirus is the important etiological agents of infectious diarrhea among children under 5 years old. Rotaviruses are divided into 10 serogroups (A–J) and each group is based on genetic properties of major structural protein VP6. We designed a novel VP6 sequence optimization to increase the expression level of this protein. Numerous factors such as codon adaptation index, codon pair bias, and guanine‐cytosine content were adapted based on Escherichiacoli codon usage. In addition, the ribosome binding site (RBS) of pET‐15b was redesigned by the RBS calculator and the secondary structure of VP6 messenger RNA was optimized in the whole length of the coding sequence. Various factors including isopropyl beta‐ d‐thiogalactoside (IPTG) concentration, temperature, and induction time were analyzed for the optimization of the best expression in E. coli by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and western blotting. The recombinant VP6 (rVP6) protein was purified by the Ni‐sepharose and then the hyperimmune sera were generated against rVP6 in rabbits. Among three different temperatures, IPTG concentrations, and postinductions, the level of rVP6 was higher at 37°C, 1 mM of IPTG, and 8 h, respectively. Also, the high expression level of rVP6 was obtained in the insoluble aggregate form (43.8 g/L). After purification, the yield of rVP6 was 10.83 g/L. The rVP6 specific antiserum was confirmed by both immunofluorescent and western blotting. The versatile sequence optimization was the reason to produce a high level of rVP6 compared to other reports and can potentially apply to produce cheaper commercial kits to diagnose serological tests and new rotavirus vaccines. Highlights: The versatile sequence optimization was the reason to produce a high level of rVP6.The high yield of rVP6 can apply to produce cheaper commercial kit.Polyclonal antibody was raised against rVP6.Immunoblotting and immunofluorescence assays showed the potency of VP6 antibody to detect RV. [ABSTRACT FROM AUTHOR]

Copyright of Agriscientia is the property of Revista AgriScientia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

During World War I (WWI), infectious diseases including tetanus were among the most important causes of death. Even though its efficacy was somewhat controversial before the war, tetanus antiserum played a key role in reducing the mortality of this disease. A vial of tetanus antiserum dating back from WWI, left behind on the French battlefield by the US Army, was borrowed from a private collection and opened. The serum contained within was characterized by orthogonal biochemical techniques to determine if any neutralizing IgGs could remain after 100 years of storage. In vitro analysis by Size Exclusion Chromatography and Serum Protein Electrophoresis suggested the presence of residual IgG. In spite of our hopes, these IgGs were not able to protect mice against tetanus toxin challenge in a neutralizing assay. Even though our results indicate the presence of remaining IgGs inside the serum, they were functionally disabled. These results show that obscurity alone is insufficient to protect IgGs from degradation over very long periods of time at room temperature. Highlights: Tetanus antiserum found its place in the therapeutic arsenal during World War I A century-old vial of tetanus antiserum was opened for biochemical and in vivo characterization Biochemical assays revealed the presence of proteins having all the characteristics of IgGs The serum was unable to protect mice against toxinic challenge [ABSTRACT FROM AUTHOR]

This review of tick paralysis caused by Ixodes holocyclus in Australia addresses the question: What are the key discoveries that have enabled effective treatment and prevention of tick paralysis in dogs and cats? Critical examination of 100 years of literature reveals that arguably only three achievements have advanced treatment and prevention of tick paralysis in animals. First, the most significant treatment advance was the commercial availability of tick antiserum in the 1930s. Hyperimmune serum currently remains the only specific anti‐paralysis tick therapy available to veterinarians in Australia. Second, advances in veterinary critical care have increased survival rates of the most severely affected dogs and cats. Critical care advancements have been enabled through specialised veterinary hospitals that can provide appropriate care 24 h a day, and advanced training of veterinarians, veterinary nurses and technicians. Third, perhaps that biggest advance of all in the last 100 years of research has been the commercial availability of the isooxazoline class of acaricidal preventatives in Australia specifically for I. holocyclus. This highly effective class of preventatives offers long duration of action, low cost, spot‐on or oral formulations and a low rate of adverse reactions. Animal owners and veterinarians now have the most useful tool of all – a reliable preventative. This review reveals the key events in research over the last 100 years and the tortuous pathway to delivering better treatment and preventative options for this enigmatic Australian parasite. [ABSTRACT FROM AUTHOR]

Three-finger toxins (3FTXs) are the most clinically relevant components in cobra (genus Naja) venoms. Administration of the antivenom is the recommended treatment for the snakebite envenomings, while the efficacy to cross-neutralize the different cobra species is typically limited, which is presumably due to intra-specific variation of the 3FTXs composition in cobra venoms. Targeting the clinically relevant venom components has been considered as an important factor for novel antivenom design. Here, we used the recombinant type of long-chain α-neurotoxins (P01391), short-chain α-neurotoxins (P60770), and cardiotoxin A3 (P60301) to generate a new immunogen formulation and investigated the potency of the resulting antiserum against the venom lethality of three medially important cobras in Asia, including the Thai monocled cobra (Naja kaouthia), the Taiwan cobra (Naja atra), and the Thai spitting cobra (Naja Siamensis) snake species. With the fusion of protein disulfide isomerase and the low-temperature settings, the correct disulfide bonds were built on these recombinant 3FTXs (r3FTXs), which were confirmed by the circular dichroism spectra and tandem mass spectrometry. Immunization with r3FTX was able to induce the specific antibody response to the native 3FTXs in cobra venoms. Furthermore, the horse and rabbit antiserum raised by the r3FTX mixture is able to neutralize the venom lethality of the selected three medically important cobras. Thus, the study demonstrated that the r3FTXs are potential immunogens in the development of novel antivenom with broad neutralization activity for the therapeutic treatment of victims involving cobra snakes in countries.

For the past few years, Edwardsiella tarda has become major problem in African catfish culture in Jambi. Detection by biochemical characteristic can lead to inaccurate result and there is a necessity to develop more specific and accurate method, one of which is Agar Gel Precipitation (AGP) method. Six samples each were collected from two African catfish farm located in District Sungai Gelam and Telanai Pura in Jambi, which was showed clinical signs of E. tarda outbreak with more than fifty percent mortality rate. Heat stable soluble antigen was prepared from 2 groups of pure culture isolated from sample for AGP test. Antiserum for test wells was antiserum of E. tarda (ATCC 15947) that have been produced by inoculating whole-cell antigen (heat-stable) and flagellar antigen (formalin-killed) in rabbit. For control positive, soluble antigen prepared from E. tarda (ATCC 15947), and control negative from Aeromonas hydophila (ATCC 35654) and Edwardsiella ictaluri (NCIMB 13272). Both antiserums were able to show positive reaction visible by the formation of specific precipitin lines between antiserum and antigen wells, and there was no precipitin reaction for negative control. In conclusion AGP method is a one of reliable technique to identify E. tarda.

Each year, millions of humans fall victim to animal envenomings, which may either be deadly or cause permanent disability to the effected individuals. The Nobel Prize-winning discovery of serum therapy for the treatment of bacterial infections (tetanus and diphtheria) paved the way for the introduction of antivenom therapies for envenomings caused by venomous animals. These antivenoms are based on polyclonal antibodies derived from the plasma of hyperimmunized animals and remain the only specific treatment against animal envenomings. Following the initial development of serum therapy for snakebite envenoming by French scientists in 1894, other countries with high incidences of animal envenomings, including Brazil, Australia, South Africa, Costa Rica, and Mexico, started taking up antivenom production against local venomous animals over the course of the twentieth century. These undertakings revolutionized envenoming therapy and have saved innumerous patients worldwide during the last 100 years. This review describes in detail the above-mentioned historical events surrounding the discovery and the application of serum therapy for envenomings, as well as it provides an overview of important developments and scientific breakthroughs that were of importance for antibody-based therapies in general. This begins with discoveries concerning the characterization of antibodies, including the events leading up to the elucidation of the antibody structure. These discoveries further paved the way for other milestones in antibody-based therapies, such as the introduction of hybridoma technology in 1975. Hybridoma technology enabled the expression and isolation of monoclonal antibodies, which in turn formed the basis for the development of phage display technology and transgenic mice, which can be harnessed to directly obtain fully human monoclonal antibodies. These developments were driven by the ultimate goal of producing potent neutralizing monoclonal antibodies with optimal pharmacokinetic properties and low immunogenicity. This review then provides an outline of the most recent achievements in antivenom research, which include the application of new biotechnologies, the development of the first human monoclonal antibodies that can neutralize animal toxins, and efforts toward creating fully recombinant antivenoms. Lastly, future perspectives in the field of envenoming therapies are discussed, including rational engineering of antibody cross-reactivity and the use of oligoclonal antibody mixtures.

Phytophthora cinnamomi Rands, a soil-borne oomycete plant pathogen, is a major threat to natural vegetation in many countries, especially Australia. The disease that it causes also has economic impacts on several agricultural and forestry systems worldwide. To investigate the virulence mechanisms of P. cinnamomi the roles of elicitins and elicitin-like proteins were examined using a susceptible model species, Lupinus angustifolius. Elicitins are 10 kDa proteins secreted by many Phytophthora species. Here, elicitins were isolated from P. cinnamomi liquid culture and separated according to their isoelectric point. A β-cinnamomin elicitin was then purified and used to develop an immunoaffinity purified antibody as a tool to examine virulence. The specificity of the β-cinnamomin antiserum was confirmed through Western-blot analysis. β-cinnamomin was found to be produced at different P. cinnamomi life stages and in inoculated plant roots by using immuno detection via confocal laser scanning microscopy. Pre-treatment of P. cinnamomi zoospores with β-cinnamomin antiserum and then inoculation of susceptible L. angustifolius roots revealed a partial loss of virulence of the pathogen. In addition, the expression of the β-cinnamomin gene at an early infection stage in susceptible L. angustifolius suggested an intrinsic role of the elicitin. Furthermore, the identification of covalently bound cell wall proteins in P. cinnamomi mycelia in this study has opened up the opportunity to investigate their role in virulence. • A protein extract from a P. cinnamomi liquid culture showed a single band at 10 kDa suggesting the presence of elicitin(s). • A purified anti-β-cinnamomin antibody recognised a single band on a western blot confirming the specificity of the antiserum. • β-cinnamomin immuno-labelling was found at different P. cinnamomi life stages and in inoculated roots. • Immunodepletion tests showed a loss of pathogen virulence towards Lupinus angustifolius confirming β-cinnamomin's role. • Elucidation of cell wall-associated proteins revealed an opportunity to examine the role of individual proteins in virulence. [ABSTRACT FROM AUTHOR]

Each year, millions of humans fall victim to animal envenomings, which may either be deadly or cause permanent disability to the effected individuals. The Nobel Prize-winning discovery of serum therapy for the treatment of bacterial infections (tetanus and diphtheria) paved the way for the introduction of antivenom therapies for envenomings caused by venomous animals. These antivenoms are based on polyclonal antibodies derived from the plasma of hyperimmunized animals and remain the only specific treatment against animal envenomings. Following the initial development of serum therapy for snakebite envenoming by French scientists in 1894, other countries with high incidences of animal envenomings, including Brazil, Australia, South Africa, Costa Rica, and Mexico, started taking up antivenom production against local venomous animals over the course of the twentieth century. These undertakings revolutionized envenoming therapy and have saved innumerous patients worldwide during the last 100 years. This review describes in detail the above-mentioned historical events surrounding the discovery and the application of serum therapy for envenomings, as well as it provides an overview of important developments and scientific breakthroughs that were of importance for antibody-based therapies in general. This begins with discoveries concerning the characterization of antibodies, including the events leading up to the elucidation of the antibody structure. These discoveries further paved the way for other milestones in antibody-based therapies, such as the introduction of hybridoma technology in 1975. Hybridoma technology enabled the expression and isolation of monoclonal antibodies, which in turn formed the basis for the development of phage display technology and transgenic mice, which can be harnessed to directly obtain fully human monoclonal antibodies. These developments were driven by the ultimate goal of producing potent neutralizing monoclonal antibodies with optimal pharmacokinetic properties and low immunogenicity. This review then provides an outline of the most recent achievements in antivenom research, which include the application of new biotechnologies, the development of the first human monoclonal antibodies that can neutralize animal toxins, and efforts toward creating fully recombinant antivenoms. Lastly, future perspectives in the field of envenoming therapies are discussed, including rational engineering of antibody cross-reactivity and the use of oligoclonal antibody mixtures. [ABSTRACT FROM AUTHOR]

To have an opportunity for reliable detection of benyvirus Beet soil-borne mosaic virus (BSBMV) and its discrimination from Beet necrotic yellow vein virus (BNYVV), an attempt to generate a specific immunoassay was made. Using a purified recombinant polypeptide corresponding to the 55 C-terminal amino acid residues of viral coat protein as antigen, a specific polyclonal antiserum against BSBMV was raised. The obtained immunoglobulin G fraction reacted with high specificity in western blot analysis with both the recombinant polypeptide expressed in Escherichia coli and total protein extract from BSBMV-infected plants. In parallel, to improve the identification of benyviruses, a duplex RT-PCR method for simultaneous detection of BNYVV and BSBMV was developed. This method is based on the combination of multiple primer sets to different pathogens into single multiplexed amplification reaction, which share single standard amplification conditions. [ABSTRACT FROM AUTHOR]

Bacterial black node, a disease of barley and wheat, is caused by Pseudomonas syringae pv. syringae (synonym pv. japonica, hereafter Psj). Here a selective medium, serine-potassium tellurite-based Psj-selective agar (SPTPsjA) was developed to isolate and detect Psj. After 5–7 days on SPTPsjA at 25 °C, characteristic black Psj colonies formed with an efficiency equal to that on potato–peptone–glucose agar. Except for P. viridiflava, P. cichorii, and some strains of P. syringae group bacteria, SPTPsjA either inhibited growth of other phytopathogenic bacteria or enabled their discrimination. To facilitate diagnosis of Psj, we prepared three antisera against three strains; in a slide agglutination test, the antisera reacted to all tested strains of Psj and failed to react to P. viridiflava and P. cichorii although some strains of other P. syringae group bacteria were indistinguishable from Psj with SPTPsjA and antisera. When barley and wheat seeds from fields in which bacterial black node had occurred were placed on SPTPsjA in the dark at 25 °C for 7 days, black colonies grew on the medium around the seeds. Among these strains, only those responsive to the above antisera were pathogenic on barley. These results show that the combination of SPTPsjA and antisera can isolate Psj from seeds of barley and wheat. [ABSTRACT FROM AUTHOR]

Abstract Motile Aeromonas septicemia (MAS), caused by new virulent Aeromonas hydrophila (vAh) strains, has been one of the major diseases in channel catfish in recent years. Previous studies showed that channel catfish developed immunity against vAh infection after immunization with the pathogen's extracellular proteins (ECP). To understand the mechanisms associated with the immunity, anti-ECP fish serum (antiserum) was analyzed in this study. Our results revealed that the antiserum elicited agglutination of both ECP and cells of vAh. Five fish proteins were identified in ECP agglutinants, including two innate immunity associated proteins (serotransferrin and rhamnose-binding lectin), two immunoglobulin M (IgM) molecules (IgM heavy chain and light chain) and a constitutively-produced protein (warm temperature acclimation protein). More than 68 vAh proteins in ECP were recognized and caused to aggregate by IgM in the antiserum. IgM was isolated from vAh cell agglutinants and the native IgM was shown to form a tetramer that was responsible for bacterial agglutination. Immunoblotting analysis indicated that the isolated native IgM was able to recognize some proteins in ECP, such as aerolysin and hemolysin (in the form of a high molecular weight heterologous polymer). Gene expression analysis by quantitative PCR showed that fish immunized with vAh ECP had more transcripts of genes coding for IgM, serotransferrin and rhamnose binding lectin than mock-immunized fish. Both innate and antibody-mediated immune responses in serum and expressed genes contributed to fish immunity upon immunization with ECP. Results of this study shed light on the versatility of vAh antigens and catfish IgM, which would help identify specific antigens for vaccine development and antigen specific antibodies in catfish. Highlights • Immunization with extracellular proteins (ECP) of virulent Aeromonas hydrophila (vAh) provides channel catfish with immunity • Anti-ECP serum elicited agglutination of both ECP and cells of vAh. • Analysis of agglutinants revealed the diversity and versatility of IgM in the anti-ECP serum. • Both innate and antibody-mediated immune responses in serum and expressed genes contributed to fish immunity. [ABSTRACT FROM AUTHOR]

Objective: To date, a commercial antibody to human Neuritin for immunoprecipitation is still limited. In this study, we aimed to develop a specific antibody for further research on the potential function of Neuritin. Methods and results: By epitope prediction of recombinant human Neuritin, the active fragment of human Neuritin that could be used as an excellent immunogen. Soluble His-tagged Neuritin was expressed and purified from Pichia pastoris. Polyclonal antibody against Neuritin was obtained by immunizing Sprague-Dawley rats with purified recombinant human Neuritin. Affinity-purified polyclonal antibody against Neuritin was characterized with indirect enzyme-linked immunosorbent assay, immunoblotting, immunoprecipitation, and immunofluorescence. The results demonstrated that the polyclonal antibody against Neuritin had been prepared successfully. The prepared antibody bound to both exogenous and endogenous Neuritin. Importantly, the anti-Neuritin polyclonal antibody could be used in immunoprecipitation assays. Conclusions: The prepared polyclonal antibody could be used in immunoprecipitation and provide researchers with a useful tool for further investigating the function and mechanism of Neuritin. [ABSTRACT FROM AUTHOR]

Contamination of avian attenuated vaccines by avian leukosis virus (ALV) is considered to be a specific horizontal transmission of ALV. Eradication of ALV contamination in vaccine virus seeds is thus a precondition of qualified vaccine production. In this study, we used the nucleoside reverse transcriptase (RT) inhibitor azidothymidine (AZT) together with monofactor antiserum against avian leukosis virus subgroup A (ALV-A) to remove ALV-A from vaccine virus seeds. Different doses of ALV-A were artificially added to the Newcastle disease virus (NDV) vaccine seeds, then the ALV-contaminated attenuated virus vaccine virus seeds were cultured in DF-1 cells. Single-drug treatment with 5 μg/mL AZT or 5% (v/v) as well as combined treatment with AZT and antiserum significantly suppressed ALV-A replication (P < 0.001) in the vaccine virus seeds. Complete absence of virus replication was observed in cells exposed to joint treatment with AZT and antiserum. The treated virus seeds met the requirements of the Chinese Ministry of Agriculture. Therefore, combined treatment with AZT and antiserum can be used to eradicate contaminating ALV-A from vaccine virus seeds, thus providing a new approach for improving vaccine safety. [ABSTRACT FROM AUTHOR]

【Objective】Human apolipoprotein B(hAPOB) is an important protein that keeps the blood lipid constant. Detecting hAPOB has a important clinical value. 【Method】In this study, the hAPOB(97-526 AA) sequence was ligated into the prokaryotic expression vector pGEX-4 T-1 to construct the pGEX-4 T-1-hAPOB(97-526 AA) prokaryotic expression plasmid which was then transferred into the host strain BL21,and an 80 kDa fusion protein was expressed with an expected molecular weight;The purified fusion protein was immunized against Hu sheep to prepare anti-hAPOB serum,and antibody titers were detected. 【Result】The antiserum produced by Huyang was the best when 0.4 mg prokaryotic expression antigen was injected each time. Indirect enzyme-linked immunosorbent assay(ID-ELISA) showed that the titer of the hAPOB antibody obtained by immunizing Hu sheep with the prokaryotic expression protein was 1∶40000. 【Conclusion】The antiserum obtained from this experiment can meet the commercial standards and lay the foundation for the subsequent application. [ABSTRACT FROM AUTHOR]

Highlights • SV2C is expressed in distinct nuclei within the brains of mouse, macaque, and human. • GABAergic and dopaminergic basal ganglia regions show enriched expression of SV2C. • Consistent cross-species SV2C expression supports translatability of rodent studies. Abstract The synaptic vesicle glycoprotein 2C (SV2C) is an undercharacterized protein with enriched expression in phylogenetically old brain regions. Its precise role within the brain is unclear, though various lines of evidence suggest that SV2C is involved in the function of synaptic vesicles through the regulation of vesicular trafficking, calcium-induced exocytosis, or synaptotagmin function. SV2C has been linked to multiple neurological disorders, including Parkinson's disease and psychiatric conditions. SV2C is expressed in various cell types—primarily dopaminergic, GABAergic, and cholinergic cells. In mice, it is most highly expressed in nuclei within the basal ganglia, though it is unknown if this pattern of expression is consistent across species. Here, we use a custom SV2C-specific antiserum to describe localization within the brain of mouse, nonhuman primate, and human, including cell-type localization. We found that the immunoreactivity with this antiserum is consistent with previously-published antibodies, and confirmed localization of SV2C in the basal ganglia of rodent, rhesus macaque, and human. We observed strongest expression of SV2C in the substantia nigra, ventral tegmental area, dorsal striatum, pallidum, and nucleus accumbens of each species. Further, we demonstrate colocalization between SV2C and markers of dopaminergic, GABAergic, and cholinergic neurons within these brain regions. SV2C has been increasingly linked to dopamine and basal ganglia function. These antisera will be an important resource moving forward in our understanding of the role of SV2C in vesicle dynamics and neurological disease. [ABSTRACT FROM AUTHOR]