Preparation and neutralization efficacy of IgY antibodies raised against Deinagkistrodon acutus venom

Background The five-paced pit viper (Deinagkistrodon acutus), endemic to China and northern Vietnam, is responsible for most snakebites in the Chinese territory. Antivenom produced from horses is the main treatment for snakebites, but it may cause numerous clinical side effects and have other disadvantages involved in their production such as the welfare of animals. The present study was conducted aiming to develop an alternative antibody (IgY) from the egg yolk of leghorn chickens immunized with snake venom. Methods IgY from the egg yolk of white leghorn chickens previously immunized intramuscularly with D. acutus venom was extracted by water, precipitated by ammonium sulfate and purified by affinity chromatographic system. IgY was identified by SDS-PAGE, ELISA and Western blot. Finally, IgY neutralization assays to test its efficacy against hemorrhagic, edema-forming and myotoxic activities of D. acutus venom were conducted on mice. Results For the first time, IgY antibodies against D. acutus venom were raised successfully in egg yolk of chickens injected with D. acutus venom multiple times. By three steps, including caprylic acid extraction, ammonium sulfate precipitation and affinity chromatography, IgY antibodies were isolated and purified from egg yolk, which exhibited a single protein band on SDS-PAGE and two bands (about 65 kDa and 35 kDa, respectively) under reducing conditions, and presented a high titer (1:40,000) tested by ELISA. Immunoblot analysis confirmed that these IgY were polyclonal antibodies since they bound to components of D. acutus venom. Furthermore, immunodiffusion assay showed that anti-D. acutus venom IgY cross-reacted with the venoms of Trimeresurus albolabris and D. saxatilis Emelianov, but did not react to the venoms of Bungarus multicinctus and Naja atra. In the neutralizing lethal assay, the median effective dose of anti-D. acutus venom IgY was 14.14 mg/kg of mouse body weight under the challenge dose (3 LD50 of D. acutus venom). In neutralizing the hemorrhagic, edema-forming and myotoxic activities of D. acutus venom, IgY showed the characteristic dose-dependent neutralization effects against all these toxic activities of D. acutus venom. Conclusion Anti-D. acutus venom IgY antibodies with high purity and titer were for the first time raised successfully in egg yolk of chickens immunized with D. acutus venom. They were effective in neutralizing the lethal effects, and the hemorrhagic, edema-forming and myotoxic acitivities of D. acutus venom. IgY could be an effective source to develop a treatment against snake bites in humans or animals in the future.(AU)

Preparation and neutralization efficacy of IgY antibodies raised against Deinagkistrodon acutus venom

Abstract Background The five-paced pit viper (Deinagkistrodon acutus), endemic to China and northern Vietnam, is responsible for most snakebites in the Chinese territory. Antivenom produced from horses is the main treatment for snakebites, but it may cause numerous clinical side effects and have other disadvantages involved in their production such as the welfare of animals. The present study was conducted aiming to develop an alternative antibody (IgY) from the egg yolk of leghorn chickens immunized with snake venom. Methods IgY from the egg yolk of white leghorn chickens previously immunized intramuscularly with D. acutus venom was extracted by water, precipitated by ammonium sulfate and purified by affinity chromatographic system. IgY was identified by SDS-PAGE, ELISA and Western blot. Finally, IgY neutralization assays to test its efficacy against hemorrhagic, edema-forming and myotoxic activities of D. acutus venom were conducted on mice. Results For the first time, IgY antibodies against D. acutus venom were raised successfully in egg yolk of chickens injected with D. acutus venom multiple times. By three steps, including caprylic acid extraction, ammonium sulfate precipitation and affinity chromatography, IgY antibodies were isolated and purified from egg yolk, which exhibited a single protein band on SDS-PAGE and two bands (about 65 kDa and 35 kDa, respectively) under reducing conditions, and presented a high titer (1:40,000) tested by ELISA. Immunoblot analysis confirmed that these IgY were polyclonal antibodies since they bound to components of D. acutus venom. Furthermore, immunodiffusion assay showed that anti-D. acutus venom IgY cross-reacted with the venoms of Trimeresurus albolabris and D. saxatilis Emelianov, but did not react to the venoms of Bungarus multicinctus and Naja atra. In the neutralizing lethal assay, the median effective dose of anti-D. acutus venom IgY was 14.14 mg/kg of mouse body weight under the challenge dose (3 LD50 of D. acutus venom). In neutralizing the hemorrhagic, edema-forming and myotoxic activities of D. acutus venom, IgY showed the characteristic dose-dependent neutralization effects against all these toxic activities of D. acutus venom. Conclusion Anti-D. acutus venom IgY antibodies with high purity and titer were for the first time raised successfully in egg yolk of chickens immunized with D. acutus venom. They were effective in neutralizing the lethal effects, and the hemorrhagic, edema-forming and myotoxic acitivities of D. acutus venom. IgY could be an effective source to develop a treatment against snake bites in humans or animals in the future.

The pilot study of radiology nursing intervention in abdominal 3-T Magnetic Resonance examination / Estudio piloto de intervención de enfermería radiológica en examen de resonancia magnética 3-t / Estudo piloto sobre a intervenção de enfermagem na radiologia em exame de ressonância magnética abdominal 3 tesla

Abstract OBJECTIVE The aim of this study was to investigate the effect of radiology nursing intervention in abdominal examination at 3-T MRI. METHOD 60 patients with abdominal diseases were divided into two groups randomly: MR nursing intervention group and control group. All the patients underwent abdominal MR examination at 3-T. The MR nursing interventions were performed in nursing intervention group. The outcomes, including one-time success rate, the ratio of diagnosable MR images and the points of image quality, were compared between these two groups. RESULTS The one-time success rates in control group and MR nursing intervention group were 66.67% and 96.67% with significant difference ( χ2 =9.017, P<0.05). The ratios of diagnosable images in the two groups were 76.67% and 96.67% with significant difference (χ2 =5.192, P<0.05). The points of MR image quality in the two groups were 1.87±0.86 and 2.33±0.55, respectively. There was significant difference between these two groups (t=-2.508, P<0.05). CONCLUSION The effective nursing intervention can make the patients cooperation better in abdominal MR examination and improve the image quality significantly.

Resumen OBJETIVO Se objetivó investigar el efecto de la intervención de enfermería radiológica en examen abdominal de IRM 3-T. MÉTODO Sesenta pacientes con enfermedades abdominales fueron divididos aleatoriamente en dos grupos: grupo intervención de enfermería RM y grupo control. Todos fueron sometidos a examen abdominal de RM 3-T. Las intervenciones de enfermería RM se efectuaron en el grupo establecido. Los resultados, incluyendo tasa de éxito en única vez, relación de imágenes de RM diagnosticables y puntos de calidad de imagen, fueron comparadas entre ambos grupos. RESULTADOS La tasa de éxito en única vez en grupo control y grupo intervención de enfermería RM fueron de 66,67% y 96,67%, expresando diferencias significativas (χ2 =9.017, P<0.05). Los puntos de calidad de imagen de RM en ambos grupos fueron 1,87±0,86 y 2,33±0,55, respectivamente. Existió diferencia significativa entre los grupos (t=-2.508, P<0.05). CONCLUSIÓN La intervención de enfermería efectiva puede mejorar la cooperación de los pacientes en examen abdominal de RM y mejorar significativamente la calidad de imagen.

Resumo OBJETIVO O objetivo deste estudo foi investigar o resultado da intervenção de enfermagem na radiologia em exame de ressonância magnética (RM) abdominal 3 Tesla. MÉTODO 60 pacientes com patologias abdominais foram aleatoriamente divididos em dois grupos: grupo de intervenção de enfermagem em RM e grupo controle. Todos os pacientes foram submetidos ao exame de RM abdominal 3 Tesla. As intervenções de enfermagem em RM foram realizadas no grupo de intervenção de enfermagem. Os resultados foram comparados entre os dois grupos, incluindo a taxa de sucesso único, o coeficiente das imagens de RM diagnosticáveis e os pontos da qualidade de imagem. RESULTADOS As taxas de sucesso único no grupo controle e no grupo de intervenção de enfermagem em RM foram 66,67% e 96,67%, com diferença significativa de (χ2 =9,017, P<0,05). Os coeficientes das imagens diagnosticáveis nos dois grupos foram 76,67% e 96,67%, com diferença significativa de (χ2 =5,192, P<0,05). Os pontos da qualidade de imagem da RM nos dois grupos foram respectivamente 1,87±0,86 e 2,33±0,55. Houve uma diferença significativa entre estes dois grupos (t=-2,508, P<0,05). CONCLUSÃO A intervenção de enfermagem eficaz pode melhorar a cooperação dos pacientes no exame de RM abdominal e melhorar consideravelmente a qualidade da imagem.

Anti-Trimeresurus albolabris venom IgY antibodies: preparation, purification and neutralization efficacy

Background Snakebite incidence in southwestern China is mainly attributed to one of the several venomous snakes found in the country, the white-lipped green pit viper Trimeresurus albolabris. Since antivenom produced from horses may cause numerous clinical side effects, the present study was conducted aiming to develop an alternative antivenom antibody (immunoglobulin Y - IgY) from leghorn chickens. Methods IgY in egg yolk from white leghorn chicken previously injected with T. albolabris venom was extracted by water, precipitated by ammonium sulfate and purified by affinity chromatographic system. IgY was identified by SDS-PAGE, ELISA and Western blot, and its neutralizing assay was conducted on mice. Results Chickens injected multiple times with T. albolabris venom elicited strong antibody responses, and from their egg yolk IgY was isolated and purified, which exhibited a single protein band on SDS-PAGE and two bands (about 65 and 35 kDa, respectively) under reduced conditions. Immunoblot analysis revealed that these IgY are polyclonal antibodies since they bind with most venom components. In the neutralizing assay, all mice survived while the ratios of IgY/venom reached up to 3.79 (50.0 mg/13.2 mg). Conclusions IgY antibody response was successfully conducted in white leghorn chicken injected with T. albolabrisvenom. IgY against T. albolabris venom was obtained for the first time, and it exhibited strong neutralizing potency on mice. These results may lay a foundation for the development of IgY antivenom with clinical applications in the future.(AU)

Anti-Trimeresurus albolabris venom IgY antibodies: preparation, purification and neutralization efficacy

Snakebite incidence in southwestern China is mainly attributed to one of the several venomous snakes found in the country, the white-lipped green pit viper Trimeresurus albolabris. Since antivenom produced from horses may cause numerous clinical side effects, the present study was conducted aiming to develop an alternative antivenom antibody (immunoglobulin Y - IgY) from leghorn chickens. Methods IgY in egg yolk from white leghorn chicken previously injected with T. albolabris venom was extracted by water, precipitated by ammonium sulfate and purified by affinity chromatographic system. IgY was identified by SDS-PAGE, ELISA and Western blot, and its neutralizing assay was conducted on mice. Results Chickens injected multiple times with T. albolabris venom elicited strong antibody responses, and from their egg yolk IgY was isolated and purified, which exhibited a single protein band on SDS-PAGE and two bands (about 65 and 35 kDa, respectively) under reduced conditions. Immunoblot analysis revealed that these IgY are polyclonal antibodies since they bind with most venom components. In the neutralizing assay, all mice survived while the ratios of IgY/venom reached up to 3.79 (50.0 mg/13.2 mg). Conclusions IgY antibody response was successfully conducted in white leghorn chicken injected with T. albolabrisvenom. IgY against T. albolabris venom was obtained for the first time, and it exhibited strong neutralizing potency on mice. These results may lay a foundation for the development of IgY antivenom with clinical applications in the future.

Design, construction, and analysis of specific zinc finger nucleases for microphthalmia - associate transcription factor

This work studied the design, construction, and cleavage analysis of zinc finger nucleases (ZFNs) that could cut the specific sequences within microphthalmia - associate transcription factor (mitfa) of zebra fish. The target site and ZFPs were selected and designed with zinc finger tools, while the ZFPs were synthesized using DNAWorks and two-step PCR. The ZFNs were constructed, expressed, purified, and analyzed in vitro. As expected, the designed ZFNs could create a double-stand break (DSB) at the target site in vitro. The DNAWorks, two-step PCR, and an optimized process of protein expression were firstly induced in the construction of ZFNs successfully, which was an effective and simplified protocol. These results could be useful for further application of ZFNs - mediated gene targeting.