Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 104
Filter
1.
Vaccimonitor (La Habana, Print) ; 30(1)ene.-abr. 2021. tab
Article in English | LILACS, CUMED | ID: biblio-1150246

ABSTRACT

Foot and mouth disease is a highly contagious viral disease of cloven-hoofed animals that has a significant economic impact on livestock. A recent outbreak was detected and recorded as exotic strain of foot and mouth disease virus SAT2 (Serotype SAT2, topotype VII, Lib-12 lineage). The emergency vaccine was produced and assessed in vivo and large number of vaccine batches were urgently needed. The present work was aimed to provide a rapid evaluation of inactivated foot and mouth disease SAT2 oily vaccine to exclude the unsatisfactory batches during emergency circumstances and to reduce time, effort and cost. The extraction of foot and mouth disease antigen content from oily adjuvanted vaccine was carried out using isopropyl myristate and benzyl alcohol methods. The extracted viral antigen was identified by foot and mouse disease serotyping ELISA and 146S content was quantified using sucrose density gradient analysis. Evaluations were carried out instantly and at 2h, 6h and 24h. The results indicated the efficiency of benzyl alcohol to breakdown the oil emulsion either MONTANIDE™ ISA 206 VG or MONTANIDE™ ISA 50 V2, while the isopropyl myristate was efficient for MONTANIDE™ ISA 50 V2 only. The identification and quantification of 146S for extracted antigen using benzyl alcohol indicated significant stable records at different time intervals for the vaccine batches, while the extraction using isopropyl myristate indicated unstable records at different time intervals. It was concluded that the evaluation of monovalent foot and mouse disease vaccine could be conducted in vitro, using serotyping ELISA and quantification of 146S for the extracted antigen, either using benzyl alcohol or isopropyl myristate (MONTANIDE™ ISA50 V2 only), with the consideration that 146S content should not less than 4 μg/mL(AU)


La fiebre aftosa es una enfermedad viral altamente contagiosa de los animales de pezuña hendida que tiene un impacto económico significativo en el ganado. Se detectó un brote reciente que se registró como causado por una cepa exótica del virus de la fiebre aftosa (serotipo SAT2, topotipo VII, linaje Lib-12). La vacuna de emergencia se elaboró y evaluó in vivo, existiendo una urgente necesidad de contar con un gran número de lotes de la misma. El presente trabajo tuvo como objetivo proporcionar una evaluación rápida de la vacuna oleosa inactivada (SAT2) contra la fiebre aftosa, para excluir los lotes insatisfactorios durante circunstancias de emergencia, reduciendo tiempo, esfuerzo y costo. La extracción del contenido de antígeno de fiebre aftosa, de la vacuna oleosa adyuvada, se llevó a cabo utilizando miristato de isopropilo y alcohol bencílico. El antígeno viral extraído se identificó utilizando un ELISA de serotipificación y se cuantificó el contenido de 146S mediante análisis de gradiente de densidad de sacarosa. Las evaluaciones se realizaron de forma instantánea y a las 2h, 6h y 24h. Los resultados indicaron la eficacia del alcohol bencílico para separar la emulsión de aceite para MONTANIDE ™ ISA 206 VG o MONTANIDE™ ISA 50 V2, mientras que el miristato de isopropilo fue eficaz para MONTANIDE™ ISA 50 V2 únicamente(AU)


Subject(s)
Animals , Enzyme-Linked Immunosorbent Assay/methods , Foot-and-Mouth Disease , Vaccines , Egypt
2.
J. venom. anim. toxins incl. trop. dis ; 27: e20200177, 2021. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1250255

ABSTRACT

The western Russell's viper (Daboia russelii) is widely distributed in South Asia, and geographical venom variation is anticipated among distant populations. Antivenoms used for Russell's viper envenomation are, however, raised typically against snakes from Southern India. The present study investigated and compared the venom proteomes of D. russelii from Sri Lanka (DrSL) and India (DrI), the immunorecognition of Indian VINS Polyvalent Antivenom (VPAV) and its efficacy in neutralizing the venom toxicity. Methods: The venoms of DrSL and DrI were decomplexed with C18 high-performance liquid chromatography and SDS-polyacrylamide gel electrophoresis under reducing conditions. The proteins fractionated were identified through nano-ESI-liquid chromatography-tandem mass spectrometry (LCMS/MS). The immunological studies were conducted with enzyme-linked immunosorbent assay. The neutralization of the venom procoagulant effect was evaluated in citrated human plasma. The neutralization of the venom lethality was assessed in vivo in mice adopting the WHO protocol. Results: DrSL and DrI venom proteomes showed comparable major protein families, with phospholipases A2 (PLA2) being the most abundant (> 60% of total venom proteins) and diverse (six protein forms identified). Both venoms were highly procoagulant and lethal (intravenous median lethal dose in mice, LD50 = 0.24 and 0.32 µg/g, for DrSL and DrI, respectively), while lacking hemorrhagic and anticoagulant activities. VPAV was immunoreactive toward DrSL and DrI venoms, indicating conserved protein antigenicity in the venoms. The high molecular weight venom proteins were, however, more effectively immunorecognized than small ones. VPAV was able to neutralize the coagulopathic and lethal effects of the venoms moderately. Conclusion: Considering that a large amount of venom can be injected by Russell's viper during envenomation, the potency of antivenom can be further improved for optimal neutralization and effective treatment. Region-specific venoms and key toxins may be incorporated into the immunization procedure during antivenom production.(AU)


Subject(s)
Animals , Poisons/toxicity , Antivenins/biosynthesis , Russell's Viper , Proteomics , Geographic Locations
3.
Acta Pharmaceutica Sinica ; (12): 1453-1459, 2021.
Article in Chinese | WPRIM | ID: wpr-887086

ABSTRACT

Galli Gigerii Endothelium Corneum (GGEC) represents digestion-promoting medicines with measurable effects and extensive clinical application. However, its effective components are not clear. The quality control index in the current edition of Chinese Pharmacopoeia is rather elementary and does not reflect its clinical efficacy. In this study, a bioassay method based on pepsin activity was proposed as a novel quality control method. With pepsin activity as the evaluation index, the extraction of GGEC was optimized and a method for the determination of biological potency was established by using the qualitative reaction parallel line method. The biological potency and consistency of 20 batches of GGEC were investigated. To provide scientific evidence in support of this bioassay method, two validation experiments were designed. One was to study the viscosity-reducing activity of a nutritional semi-solid paste after adding GGEC samples with differing potency. The other was to correlate the gastric residual rate in mice and pepsin activity with the alcohol soluble extract content. The results showed that the optimal preparation method was to dilute crude powder of GGEC with 50 volumes of water and subject to ultrasonic extraction at 300 W and 40 kHz for 0.5 h. The shape of the dose-response curve was similar to that of the positive control drug multienzyme tablets and the precision, intermediate precision and repeatability met the methodology requirements. The results showed that the potency of 20 batches of samples ranged from 13.49 to 34.69 U·mg-1, with an average value of 22.21 U·mg-1. The validation experiment demonstrated that the effect of reducing the viscosity of the nutrient paste became more significant as GGEC sample potency increased. The correlation coefficient R of gastric residual rate with pepsin potency and alcohol soluble extract content was 0.867 and 0.518, respectively, which indicated that the pepsin potency was highly correlated with in vivo activity. This study shows that a bioassay method based on pepsin activity is reliable and reproducible for GGEC and could provide reference method for the quality evaluation of other digestant herbs.

4.
Article | IMSEAR | ID: sea-209639

ABSTRACT

Plant products have been tested as insecticides against mosquitoes as they are promising candidates to replace conventional insecticides. This study was carried out to evaluate the larvicidalpotential of ethanol extract of the aerial parts of Diplazium esculentumagainst Anopheles gambiaeand Culex quinquefasciatus. Ethanol extract of the aerial parts of D. esculentumwas screened for its phytochemical constituents and used for larvicidal assay. A stock solution of the extract (5g in Original Research Article Umohata et al.; IJTDH, 41(3): 40-47, 2020; Article no.IJTDH.5566841100ml of water) was prepared. From the stock solution, 0.45, 0.60, 0.75, 0.90 and 1.05%w/v concentrations of the extract were obtained for the study. Each concentration of the extract had 3 replicates. The control was also replicated. Twenty (20) third instar larvae each of Anopheles gambiae andCulex quinquefasciatuswere separately exposed to each extract concentration for a duration of 48 hours. Larval nutrient was added to each experimental set up. Observations were made after 24 and 48 hours exposure period.Phytochemical screening revealed the presence of some plant metabolites. Mortality of larvae exposed to the extract increased with increased concentration and exposure time. This study revealed a differential susceptibility of larvae of the two mosquito species to the extract as evident by the 24h LC50values obtained which were 0.355 and 2.468%w/v for An. gambiaeand Cx.quinquefasciatus respectively. Exposure of An. gambiaelarvae to the extract resulted in 100% mortality even with the least concentration of 0.45%w/v after 48 hours exposure period while the highest concentration of extract (1.05%w/v) resulted in 53.33% mortality of Cx.quinquefasciatuslarvae, after an exposure period of 48 hours. Results obtained from this study suggest that the aerial parts of D. esculentumif further explored would be useful in the control of An. gambiae andCx.quinquefasciatus.

5.
Biomédica (Bogotá) ; 40(1): 43-54, ene.-mar. 2020. tab, graf
Article in Spanish | LILACS | ID: biblio-1089104

ABSTRACT

Introducción. Las diferencias en la activación muscular de las porciones lateral y medial del cuádriceps durante la ejecución del salto, pueden convertirse en un factor de riesgo neuromuscular al aumentar el valgo dinámico de rodilla y, en consecuencia, el riesgo de lesión del ligamento cruzado anterior de la rodilla. Objetivo. Determinar la diferencia en la activación de los vastos lateral y medial del cuádriceps mediante electromiografía de superficie durante el salto con una sola pierna en los planos sagital y frontal en mujeres deportistas. Materiales y métodos. Se hizo un estudio cuantitativo de corte transversal con la participación de 64 mujeres deportistas a quienes se les tomaron las medidas antropométricas. Se hicieron pruebas de salto vertical y lateral con una sola pierna evaluados mediante la activación electromiográfica de los vastos medial y lateral, y la valoración de la flexibilidad de la banda iliotibial. Resultados. Se encontró una relación estadísticamente significativa (p≤0,05) entre el índice de masa corporal, el porcentaje de grasa y la potencia en los saltos verticales con una sola pierna. Se encontró, asimismo, significación estadística (p≤0,05) por una mayor activación del vasto lateral en el salto vertical con la pierna derecha y en el salto lateral con las dos piernas. Conclusión. Las deportistas presentaron diferencias en la activación de los cuádriceps, siendo mayor la activación del vasto lateral en la mayoría de los saltos con una sola pierna tanto en el plano sagital como en el frontal, lo cual puede contribuir a un aumento del riesgo de lesión de rodilla en la práctica deportiva.


Introduction: Differences in the lateral and medial portions from quadriceps muscular activation during the execution of the jump can become a neuromuscular risk factor raising knee dynamic valgus and increasing the risk for anterior cruciate ligament injury. Objective: To determine the difference in the activation of the lateral and medial vastus using surface electromyography during single-leg jumps in the sagittal and frontal plane in female athletes. Materials and methods: This was a quantitative, cross-sectional study. A total of 64 female athletes participated. We carried out anthropometric measurements, vertical and lateral single-leg jump tests with the evaluation of vastus medialis and lateral electromyographic activation during the tests and iliotibial band flexibility assessment. Results: There was a statistically significant relationship (p≤0.05) between the body mass index, fat percentage, and vertical single-leg jump power. We also found statistical significance (p≤0.05), with greater activation of the lateral vastus in the right lower limb vertical jump and in the lateral jump in both lower limbs. Conclusion: The athletes presented differences in quadriceps activation, showing higher lateral vastus activation in most of the single-leg jumps, both in the sagittal and frontal plane. This may increase the risk for suffering knee injuries during sports practice.


Subject(s)
Sports , Women , Anterior Cruciate Ligament , Primary Prevention , Potency , Electromyography , Knee , Muscles
6.
Article in Chinese | WPRIM | ID: wpr-846562

ABSTRACT

Objective: To establish a biological potency assay for Xiaojin Pills against platelet aggregation in vitro, evaluate the quality consistency of Xiaojin Pills, and screen traditioanal Chinese medicines which play the role of promoting blood circulation in Xiaojin Pills. Methods: Xiaojin Pills and ten Chinese medicines [artificial musk, Momordica cochinchinensis, Aconitum kusnezoffii, Liquidambar formosana, Boswellia carterii, Commiphora myrrha, Faeces Trogopterori, Angelica sinensis, Pheretima aspergillum, Fragrant Ink] in its formula were extracted by ultrasound in 40% methanol. The antiplatelet aggregation rate of the extract was measured by platelet aggregation meter. The platelet-rich plasma (PRP) and platelet-poor plasma (PPP) were prepared from abdominal aorta of rats. The platelet aggregation was induced by adenosine diphosphate (ADP). With sodium ferulate as a standard reference material, the biological potency of antiplatelet aggregation of Xiaojin Pills was calculated by the simplified probit principle. Results: The results showed that the biological potency of Xiaojin Pills was between 0.598 and 1.338 U/mg among different manufacturers and batches. In Xiaojin Pills group, Pheretima, Faeces Trogopterori, and Momordicae Semen had stronger inhibitory effects on platelet aggregation with inhibition rates of 70.87%, 31.83% and 67.52%, respectively. Conclusion: The quality consistency of Xiaojin Pills from different manufacturers and batches is poor, and Pheretima, Faeces Trogopterori, and Momordicae Semen may be the key drugs for Xiaojin Pills to play the role of promoting blood circulation.

7.
Chinese Pharmaceutical Journal ; (24): 133-140, 2020.
Article in Chinese | WPRIM | ID: wpr-857800

ABSTRACT

OBJECTIVE: To establish a high performance liquid chromatography combined with pulsed amperometric detection(HPLC-PAD)method for determination of potency of neomycin sulfate. METHODS: An improved HPLC-PAD method from EP method for determination of the content and related substances of neomycin sulfate was established and validated. The study of impurity profile of neomycin sulfate was completed by LC-IT-TOF method with the help of on-line desalination using a suppressor; and the main components in neomycin sulfate were clarified combining the RESULTS of impurity profile and minimum inhibitory concentrations of the main components and impurities. The semi-preparative liquid chromatography-evaporative light scattering detector(ELSD) was self-assembled, highly purified neomycin B and neomycin C were prepared and their structural confirmation was also conducted. The contents of highly purified neomycin B and neomycin C were determined by means of mass balance method. The potencies of highly purified neomycin B and neomycin C were determined by three-dose antibiotic microbial assay and the conversion factors between contents of neomycin B and neomycin C and their potencies were calculated separately and then a formula for the calculation of potency of neomycin sulfate from the content of main components of neomycin B and neomycin C was obtained.At last, a verification experiment for the accuracy of the conversion factor and the formula were designed and a serial of tests were carried out to investigate the interaction and the verification for the actual sample. RESULTS: The improved HPLC-PAD method was superior to the European Pharmacopoeia method in the separation ability and stability, and was suitable for accurate quantification of various components of neomycin sulfate and related substance inspection. The successful removal of trifluoroacetic acid in the mobile phase by the technology of desalination on-line using a suppressor broke a new way for the study of impurity profile of aminoglycoside such as neomycin sulfate. Combining the impurity profile with the RESULTS of MIC it was clarified that the main activity components of neomycin sulfate were neomycin B and neomycin C. Highly purified neomycin B and neomycin C were successfully prepared. A conversion factor for the transition from potency to purity of neomycin sulfate was obtained through experiments and calculations and was verified successfully. CONCLUSION: It is feasible to replace the microbial assay by HPLC-PAD method for determining the potency of neomycin sulfate.

8.
Chinese Journal of Biotechnology ; (12): 1378-1385, 2020.
Article in Chinese | WPRIM | ID: wpr-826839

ABSTRACT

Listeria monocytogenes (Lm) is zoonotic pathogen that can cause listeriosis, and vaccine is one of the effective methods to prevent this pathogen infection. In this study, we developed a novel vaccine that is a mixture of inactivated bacteria and Montanide™ ISA 61 VG, a mineral oil adjuvant, and evaluated the safety and immune response characteristics of this vaccine. The mice immunized with the ISA 61 VG adjuvant had high safety, and it could induce significantly higher titer of anti-listeriolysin O (LLO) antibody and higher value of IgG2a/IgG1 ratio compared with the group without the adjuvant. In particular, it could provide 100% immune protection against lethal doses of Lm challenge in mice. In summary, ISA 61VG adjuvant significantly enhanced the ability of inactivated listeria vaccine to induce humoral and cellular immune responses, thereby enhanced the protective immune response in the host, and it is a potential vaccine candidate for the prevention of Lm infection in humans and animals.


Subject(s)
Adjuvants, Immunologic , Pharmacology , Animals , Hemolysin Proteins , Allergy and Immunology , Pharmacology , Immunity, Cellular , Listeria monocytogenes , Allergy and Immunology , Listeriosis , Mice , Mice, Inbred BALB C , Vaccines, Inactivated , Allergy and Immunology
9.
Psicol. soc. (Online) ; 32: e175161, 2020.
Article in Portuguese | LILACS-Express | LILACS, INDEXPSI | ID: biblio-1135932

ABSTRACT

Resumo Neste texto, amparados pelos conceitos de Deleuze e utilizando-nos do método cartográfico, que consiste no acompanhamento de processos de uma realidade, objetivamos compartilhar um momento de nossa pesquisa-intervenção centrada na produção estética dos corpos em escolas, realizada em escola municipal de EJA. Focar-nos-emos no acompanhamento do caso de uma estudante, a qual angustiava aos educadores por ser entendida como sujeito de educação especial e vítima de abuso sexual. Lançando mão da noção deleuzeana de ética para acompanhar esse caso, discutiremos como se dá a construção de uma educação ética, a qual pretende aumentar a potência de conhecer baseada na produção de encontros alegres. Veremos que a estudante pôde aprender sobre suas potências à medida que a escola, sem vitimizá-la, agenciou encontros baseados naquilo que seu corpo podia.


Resumen En este texto, con el apoyo de los conceptos de Deleuze y del método cartográfico, que consiste em acompañar los procesos de una realidad, nuestro objetivo es compartir un momento de nuestra pesquisa-intervención, que se centró en la producción estética de los cuerpos en escuelas, realizada en una escuela municipal de EJA. Nos centraremos en el acompañamiento del caso de una estudiante que afligía a los educadores por ser entendida como sujeto de educación especial y víctima de abuso sexual. Usando el concepto deleuzeano de ética para seguir este caso, discutiremos cómo se da la construcción de una educación ética, cuyo objetivo es aumentar la potencia del conocer basada en la producción de encuentros alegres. Veremos que la estudiante ha podido aprender sobre sus potencias a la medida que la escuela, sin victimizarla, agenció encuentros basados en lo que su cuerpo pudiera.


Abstract In this paper, supported by Deleuze´s concepts and using the cartographic method, which consists in following the processes of a reality, we aim to share a moment of our intervention-research focused on aesthetic bodies production in schools, which was developed in a public school of youth and adult education. We focused on monitoring the case of a student, who distressed educators for being understood as a special education subject and victim of sexual abuse. Making use of the Deleuzian notion of ethics to accompany this case, in this paper we discuss the production of an ethical education, which aims to increase the power of knowing based on the promotion of cheerful gatherings. In this process, it can be noted that the student was able to learn about her potentials as the school promoted meetings based on what her body could, without victimizing her.

10.
Article | IMSEAR | ID: sea-205273

ABSTRACT

Background: Corticosteroids are widely prescribed drugs in dermatology. Rational prescribing of steroids is important for best therapeutic outcome at lowest possible dose. A study was carried out at a tertiary care teaching hospital in order to evaluate the use of corticosteroids which provided a picture of trends in the usage of corticosteroids in dermatology at that set-up. Materials and Methods: This prospective, observational study was carried out in department of dermatology for 1 year after ethical approval. Data was analysed for parameters related to corticosteroids, their potency, WHO drug prescribing indicators, effectiveness as well as effects of corticosteroids on quality of life of patients. Statistical analysis was done using Microsoft Excel Office 365. Results: In the 223 patients, 44.84% patients belonged to 21-40 years age group. Mostcommon indication was eczema in 29.15% cases. Topical betamethasone (25.11%) and oral prednisolone (20.17%) were most frequently prescribed. 95/140 topical steroids prescribed were super highly potent. Among concomitant drugs, a majority of 38% were antihistaminics. Degree of polypharmacy showed 04 drugs in a majority (43.15%) of prescriptions. Only 6.27% drugs were prescribed by generic name. Conclusion: Corticosteroids were beneficial to a large no. of patients. Initial usage of low potency steroids topically wherever possible can be emphasized. WHO drug prescribing indicators analysis indicated the need to adhere to WHO guidelines as well as prescribing drugs by generic name. To maintain a balance between judicious use and frequent abuse with corticosteroid is important along with physician’s vigilance and patient education.

11.
Indian J Med Microbiol ; 2019 Mar; 37(1): 95-98
Article | IMSEAR | ID: sea-198842

ABSTRACT

There is a need of a relatively simple and inexpensive method for the determination of relative potency of various generic brands of antibiotics in comparison to original products. The current study describes an agar diffusion method which can be performed in any microbiology laboratory, is cheap (costs $2 per test) and its results can be available after overnight incubation. The results show that neither all generics are reliable nor are all generic antibiotics of poor quality.

12.
Article in English | WPRIM | ID: wpr-785829

ABSTRACT

BACKGROUND AND OBJECTIVES: Most studies in cardiac regeneration have explored bone marrow mesenchymal stem cells (BM-MSC) with variable therapeutic effects. Amniotic fluid MSC (AF-MSC) having extended self-renewal and multi-potent properties may be superior to bone marrow MSC (BM-MSC). However, a comparison of their cardiomyogenic potency has not been studied yet.METHODS: The 5-azacytidine (5-aza) treated AF-MSC and BM-MSC were evaluated for the expression of GATA-4, Nkx2.5 and ISL-1 transcripts and proteins by quantitative RT-PCR and Western blotting, respectively as well as for the expression of cardiomyogenic differentiation markers cardiac troponin-T (cTNT), beta myosin heavy chain (βMHC) and alpha sarcomeric actinin (ASA) by immunocytochemistry.RESULTS: The AF-MSC as compared to BM-MSC had significantly higher expression of GATA-4 (183.06±29.85 vs. 9.80±0.05; p<0.01), Nkx2.5 (8.3±1.4 vs. 1.82±0.32; p<0.05), and ISL-1 (39.59±4.05 vs. 4.36±0.39; p<0.01) genes as well as GATA-4 (2.01±0.5 vs. 0.6±0.1; p<0.05), NKx2.5 (1.9±0.14 vs. 0.8±0.2; p<0.01) and ISL-1 (1.7±0.3 vs. 0.9±0.1; p<0.05) proteins. The AF-MSC also had significantly elevated expression of cTNT (5.0×10⁴±0.6×10⁴ vs. 3.5 ×10⁴±0.8×10⁴; p<0.01), β-MHC (15.7×10⁴±0.9×10⁴ vs. 8.2×10⁴±0.6×10⁴; p<0.01) and ASA (18.6×10⁴±4.9×10⁴ vs. 13.1×10⁴±3.0×10⁴; p<0.05) than BM-MSC.CONCLUSIONS: Our data suggest that AF-MSC have greater cardiomyogenic potency than BM-MSC, and thus may be a better source of MSC for therapeutic applications in cardiac regenerative medicine.


Subject(s)
Actinin , Amniotic Fluid , Antigens, Differentiation , Azacitidine , Blotting, Western , Bone Marrow , Female , Humans , Immunohistochemistry , Mesenchymal Stem Cells , Regeneration , Regenerative Medicine , Therapeutic Uses , Troponin T , Ventricular Myosins
13.
Article in Chinese | WPRIM | ID: wpr-756232

ABSTRACT

Objective To validate a cell infection-based quantitative RT-PCR for evaluating the potency of rotavirus vaccine. Methods According to the ICH ( the International Council for Harmonization) Harmonised Tripartite Guideline, the method was validated for its specificity, accuracy, precision, linearity and robustness. Results The method had good specificity as it could only amplify and detect the corre-sponding type of rotavirus strain. The recovery rates for determining the potency against rotaviruses of G2, G3 and G4 types were 97% to 108%. The percent coefficient of variation ( CV) of both intra-plate and in-ter-plate precision was≤2. 62%, while the intraday and interday CV was≤1. 76% and≤2. 27%, respec-tively. The CV between the two experimenters was≤7. 68%. The linearity range of the method was 4. 4-6. 5 UI for G2 type rotavirus, 3. 9-8. 3 UI for G3 type and 3. 5-8. 1 UI for G4 type. Good robustness was observed using the cells of 140 to 160 generations. Conclusions The cell infection-based quantitative RT-PCR was shown to have satisfactory specificity, accuracy, precision, linearity and robustness, suggesting that it was a suitable method for evaluating the potency of multivalent rotavirus live vaccines.

14.
Chinese Pharmaceutical Journal ; (24): 1099-1103, 2019.
Article in Chinese | WPRIM | ID: wpr-857977

ABSTRACT

OBJECTIVE: To study the algorithm of four-parameter logistic regression and apply the model in the potency calibration of bacterial endotoxin standard. METHODS: The potency of endotoxin reference standard was estimated by a novel monocyte activation test,HL60-IL6 assay.The endotoxin national standard was set as standard, while the endotoxin working standard was set as test. The four-parameter logistic model was fitted by the method of nonlinear least squares when observations of standard or sample solutions were plotted against endotoxin concentration. The algorithm was performed by C Language, the RESULTS: of which were compared by those of softmax, R Language as well as Microsoft Office Excel. RESULTS The estimated parameters as well as estimated potency were parallel between the four statistical tools. The goodness-of-fit were all over 0.99.The variance was within 0.1%. CONCLUSION: The four-parameter logistic model can be established by the four statistical tools. Researchers can select a suitable tool according to demands.

15.
Chinese Pharmaceutical Journal ; (24): 2010-2017, 2019.
Article in Chinese | WPRIM | ID: wpr-857819

ABSTRACT

OBJECTIVE: To develop a novel optimization and validation method based on design of experiment(DOE) for the antibody-dependent cell-mediated cytotoxicity (ADCC) potency of anti-programmed cell death 1 and anti-programmed cell death-ligand 1 (PD-1/PD-L1) monoclonal antibodies using reporter genes. METHODS: Jurkat-hFcγRIIIa-NFAT transgenic cell line was used as effector cells, 293FT-PD-1 cell line and CHO-PD-L1 cell line were used as target cells, respectively. The ADCC potency for anti-PD-1/PD-L1 monoclonal antibodies was detected with Luciferase detection system (BrightGloTM Luciferase Assay system), then the method was optimized and validated based on DOE. RESULTS: The anti-PD-1/PD-L1 monoclonal antibodies showed a dose-response relationship and the determination result complied with the following four-parameter equation: y=(A-D)/+D. The method was optimized and the testing parameters were determined as follows: the working concentration of anti-PD-1 monoclonal antibody was 10 000 ng•mL-1 to 4.833 ng•mL-1 and that of anti-PD-L1 was 2 000 ng•mL-1 to 0.488 ng•mL-1, the ratio of effector cells and target cells for anti-PD-1/PD-L1 monoclonal antibodies were 6:1 and 3:1, and the induction time for both of these antibodies was 20 h. The method possessed good specificity. The recovery rate test samples in the four different dilution groups were determined for 3 times, and the results showed that the relative potencies of anti-PD-1 monoclonal antibody were (51.74±2.22)%, (77.12±3.14)%, (118.71±2.83)% and (156.20±12.99)%, and the recoveries of which were (103.49±4.44)%, (102.83±4.19)%, (94.96±2.26)% and (104.14±8.66)%, respectively. While as for anti-PD-L1 monoclonal antibody, the relative potencies were (54.32±4.75)%, (75.24±4.25)%, (127.40±2.43)%, (156.82±3.27)% and the recoveries were (108.64±9.51)%, (100.33±5.67)%, (101.92±1.94)% and (104.55±2.18)%, respectively. The RSDs of the above results were all less than 10%. CONCLUSION: A novel optimization and validation method based on DOE for detecting ADCC potency of anti-PD-1/PD-L1 mAb is successfully developed. This detecting method based on reporter gene shows high specificity, good reproducibility and high accuracy, and might be used in the evaluation of ADCC potency of anti-PD-1/PD-L1 mAb.

16.
Article in Chinese | WPRIM | ID: wpr-850800

ABSTRACT

Quality marker (Q-marker) is one of the new directions in the research of quality control of Chinese materia medica. The Q-marker can be either quality chemomarker (Q-chemomarker) or quality biomarker (Q-biomarker). The establishment of a good Q-biomarker can improve the linkage of Chinese medicine quality control methods and standards with clinical safety and effectiveness, at the same time considering the operability of quality control. At present, the FDA of China and the United States have adopted biological evaluation methods as an important part of quality control of Chinese medicines and botanicals medicines. Many companies and researchers are also actively exploring and developing methods for the evaluation of quality of Chinese materia medica. This paper further enlarges and expands the methods and indicators of biological evaluation of Chinese materia medica, and summarizes and analyzes relevant research examples from the perspective of Q-biomarker. We hope that it would promote the richness and development of the theoretical system of Chinese medicine quality markers, so as to provide a reference to solve the practical problems for the quality control and evaluation of Chinese materia medica.

17.
Acta Pharmaceutica Sinica ; (12): 2141-2148, 2019.
Article in Chinese | WPRIM | ID: wpr-780352

ABSTRACT

To explore the application of an effect-constituents index (ECI) for the quality evaluation of rhubarb, we carried out the simultaneous determination of 12 chemical components by ultra-performance liquid chromatography and used the ICR mouse constipation model to determine the diarrhea biopotency of these 12 components. With the diarrhea biopotency of sennoside A as a reference, the diarrhea biopotency weight coefficient of each chemical component was obtained. A multi-component chemical quantitative analysis combined with the biopotency weight coefficients for rhubarb was developed, named the diarrhea ECI. Animal experiment ethics requirements were approved by the Animal Experimental Ethics Committee of the 302 Hospital of the People's Liberation Army (Grant Number: IACUC-2015-012). The results showed that there were significant differences in the content of the 12 chemical components in different batches of processed products of rhubarb. Especially worthy of attention was the content of aloe-emodin-8-O-β-D-glucoside in sample Rh03, nearly 40-fold higher than that in Rh07 (4.79 vs 0.12 mg·g-1), and the content of rhein-8-O-β-D-glucoside in sample Rh03, nearly 45 times higher than that in Rh07 (3.56 vs 0.08 mg·g-1). The actual measured diarrhea biopotencies of the 12 chemical components ranged from 61.65 ± 4.28 to 233.84 ± 5.58 U·mg-1. The calculated diarrhea effect-constituents indices of 16 rhubarb samples ranged from 1.07 (Rh15) to 19.38 (Rh03), and the actual measured diarrhea biopotencies of 16 rhubarb samples based the ICR mouse constipation model ranged from 23.84 U·g-1 (Rh16) to 310.94 U·g-1 (Rh05). The correlation between the diarrhea ECIs and the actual measured diarrhea biopotencies of 16 rhubarb samples was good (r = 0.969 5), suggesting that the diarrhea effect-constituents indices may be the most suitable for evaluating the quality of different rhubarbs with regard to diarrhea.

18.
Acta Pharmaceutica Sinica ; (12): 2149-2154, 2019.
Article in Chinese | WPRIM | ID: wpr-780333

ABSTRACT

We established a quality evaluation method for Shuanghuanglian preparations based on an effect-constituent index (ECI), which is guided by the clinical efficacy of Shuanghuanglian and a dose-efficacy correlation. An HPLC method was used to establish the quantitative fingerprint of Shuanghuanglian from different manufacturers and to determine the content of 10 fingerprint components, including baicalin, chlorogenic acid, forsythin, galuteolin, wogonin, forsythoside A, luteolin, caffeic acid, baicalein, and scutellarin. Using Staphylococcus aureus as biological model, the potency of Shuanghuanglian preparations was determined by antibiotic microbial assay. Using the method of PLC-DA, the efficacious antibacterial components were screened by "dose-efficacy" correlation analysis. According to the antibacterial potency and content of the antibacterial ingredients, combined with the method of the custom weight coefficient, ECI was calculated and verified. The results show that the antibacterial ECI can facilitate evaluation of the efficacy of Shuanghuanglian based on the composition of its contents, providing a new method for the quality control of traditional Chinese medicine.

19.
Acta Pharmaceutica Sinica ; (12): 2178-2183, 2019.
Article in Chinese | WPRIM | ID: wpr-780326

ABSTRACT

The objective of this work was to develop a bioassay to quantify the antiplatelet aggregation activity of hirudo for quality evaluation and control. Antithrombin activity of hirudo extracted by high temperature decoction was determined by thrombin titration. Antiplatelet aggregation activity of hirudo was determined through pharmacodynamic experiments in vitro and in vivo using a bioassay we developed for quantifying inhibition of platelet aggregation. Methodological investigation was carried out and the titers of 12 batches of hirudo samples were determined. During the experiment, the disposal of animals is in accordance with the ethical standards of animal experiments. The results showed that the antithrombin activity of hirudo decocted at high temperature decreased significantly and almost lost its activity. Hirudo inhibited platelet aggregation and results in vivo and in vitro were consistent. These assays were employed to test 12 batches of hirudo. The results demonstrated that the biopotency of 12 batches was 113.49, 96.13, 121.22, 127.33, 83.48, 108.72, 131.41, 127.95, 76.90, 126.27, 132.89 and 573.53 U·mg-1. The method was reliable and reproducible and can be used to assess the quality of hirudo.

20.
Article in English | WPRIM | ID: wpr-761793

ABSTRACT

In drug discovery or preclinical stages of development, potency parameters such as IC₅₀, K(i), or K(d) in vitro have been routinely used to predict the parameters of efficacious exposure (AUC, C(min), etc.) in humans. However, to our knowledge, the fundamental assumption that the potency in vitro is correlated with the efficacious concentration in vivo in humans has not been investigated extensively. Thus, the present review examined this assumption by comparing a wide range of published pharmacokinetic (PK) and potency data. If the drug potency in vitro and its in vivo effectiveness in humans are well correlated, the steady-state average unbound concentrations in humans [C(u_ss.avg) = f(u)·F·Dose/(CL·τ) = f(u)·AUCss/τ] after treatment with approved dosage regimens should be higher than, or at least comparable to, the potency parameters assessed in vitro. We reviewed the ratios of C(u_ss.avg)/potency in vitro for a total of 54 drug entities (13 major therapeutic classes) using the dosage, PK, and in vitro potency reported in the published literature. For 54 drugs, the C(u_ss.avg)/in vitro potency ratios were < 1 for 38 (69%) and < 0.1 for 22 (34%) drugs. When the ratios were plotted against f(u) (unbound fraction), “ratio < 1” was predominant for drugs with high protein binding (90% of drugs with f(u) ≤ 5%; i.e., 28 of 31 drugs). Thus, predicting the in vivo efficacious unbound concentrations in humans using only in vitro potency data and f(u) should be avoided, especially for molecules with high protein binding.


Subject(s)
Drug Discovery , Humans , In Vitro Techniques , Plasma , Protein Binding
SELECTION OF CITATIONS
SEARCH DETAIL