Semi-perfusion cultures of suspension MDCK cells enable high cell concentrations and efficient influenza A virus production

Control and prevention of rapid influenza spread among humans depend on the availability of efficient and safe seasonal and pandemic vaccines, made primarily from inactivated influenza virus particles. Current influenza virus production processes rely heavily on embryonated chicken eggs or on cell culture as substrate for virus propagation. Today’s efforts towards process intensification in animal cell culture could innovate viral vaccine manufacturing using high-yield suspension cells in high cell density perfusion processes. In this work, we present a MDCK cell line adapted to grow as single cell suspension with a doubling time of less than 20 h, achieving cell concentrations over 1 × 10⁷ cells/mL in batch mode. Influenza A virus titer obtained in batch infections were 3.6 log₁₀(HAU/100 µL) for total- and 10⁹ virions/mL for infectious virus particles (TCID₅₀), respectively. In semi-perfusion mode concentrations up to 6 × 10⁷ cells/mL, accumulated virus titer of 4.5 log₁₀(HAU/100 µL) and infectious titer of almost 10¹⁰ virions/mL (TCID₅₀) were possible. This exceeds results reported previously for cell culture-based influenza virus propagation by far and suggests perfusion cultures as the preferred method in viral vaccine manufacturing.     

Exercise-induced laryngeal obstruction: From clinical examination to continuous laryngoscopy during exercise

The diagnosis of exercise-induced laryngeal obstruction (EILO) should be suspected when exercise triggers inspiratory stridor. EILO is common in adolescent populations and has a significant impact on sports. Identification of laryngeal obstruction during continuous laryngoscopy during exercise (CLE) is the gold standard diagnostic tool for this disorder, which is not widely known in France. The challenge faced by otolaryngologists is to identify, among patients referred by pulmonologists or sports physicians, those with exercise-induced inspiratory symptoms or poorly controlled exercise-induced asthma, in whom a diagnosis of EILO is strongly suspected. Laryngoscopy at rest may reveal a laryngeal, glottic or supraglottic abnormality predictive of obstruction at increased inspiratory airflow. When pulmonary function tests are normal or in the case of failure of treatment of exercise-induced asthma, the otolaryngologist must complete the examination by a CLE test to confirm the diagnosis of EILO and identify the site of obstruction. This examination is well tolerated, minimally invasive and allows identification of the site of airflow obstruction, allowing specific conservative or surgical treatment. This technical note describes in detail clinical examination and CLE testing in patients with suspected EILO.     

Dorsal suspension for Morton’s neuroma: A comparison with neurectomy

BackgroundThe purpose of this study was to investigate and compare the clinical outcomes of dorsal suspension with those of neurectomy for the treatment of Morton’s neuroma.MethodsWe conducted a retrospective study of dorsal suspension and neurectomy group. The dorsal suspension was performed by dorsal transposition of neuroma over the dorsal transverse ligament after neurolysis. The visual analog scale (VAS), the Foot and Ankle Ability Measure (FAAM), postoperative satisfaction, and complications were evaluated.ResultsBoth groups reported significant pain relief, and there were no significant differences between the groups with respect to postoperative pain. The postoperative FAAM outcomes showed no significant between-group differences. Satisfaction analysis showed ‘excellent’ and ‘good’ results in the dorsal suspension and neurectomy groups (95% and 77.7%, respectively). Complications of numbness and paresthesia reported in the dorsal suspension group (5% and 5%, respectively) were significantly fewer than those of neurectomy group (61.1% and 33.3%, respectively) (both, p < .05).ConclusionsWith its favorable results, dorsal suspension can be another operative option for the treatment of Morton’s neuroma.Level of Evidence: Level III, retrospective comparative case series.     

LC-HR-MS/MS法鉴定头孢丙烯干混悬剂中的未知杂质

目的:检测头孢丙烯干混悬剂中的未知杂质,并对其进行结构鉴定。方法:采用高效液相色谱-串联高分辨质谱法检测并鉴定头孢丙烯干混悬剂中的未知杂质。色谱柱为Thermo HyPURITYTMC18,流动相为乙腈-0.013%甲酸水溶液(梯度洗脱),检测波长为230 nm,流速为1.0 m L/min,柱温为40℃,进样量为20μL;以电喷雾离子源行正离子全扫描,扫描范围为质荷比(m/z)100~1500,喷雾电压为3.8 kV,金属毛细管温度为320℃,鞘气压力为60 Arb,辅助气压力为10 Arb,喷雾温度为280℃。结果:在该色谱条件下,杂质K的检测限为0.202μg/mL,精密度、重复性试验的RSD均小于4%。杂质K附近发现3个未知杂质,且互为异构体,离子保留时间为17.83~19.31 min,二级母离子均为m/z 436.1500[M+H]+,可能为头孢丙烯开环、脱水后的产物。结论:本方法检测出头孢丙烯干混悬剂中杂质K附近的3个未知杂质。     

多索茶碱联合布地奈德混悬液雾化吸入对哮喘患者炎性因子及肺功能的影响

目的探讨多索茶碱联合布地奈德混悬液雾化吸入治疗哮喘的临床效果。方法选择我院2018年5月至2019年4月收治的哮喘患者114例,随机分为两组各57例。对照组采用布地奈德混悬液雾化吸入治疗,观察组在此基础上采用多索茶碱治疗。观察两组的临床疗效、炎性因子指标(IL-4、 IFN-γ)、肺功能指标(FVC、 FEV1)及不良反应情况。结果观察组的治疗总有效率为92.98%,高于对照组的78.95%(P <0.05)。治疗后,观察组的IL-4水平低于对照组,IFN-γ、 FEV1、 FVC水平均高于对照组,差异有统计学意义(P <0.05)。两组的不良反应发生率比较,差异无统计学意义(P>0.05)。结论多索茶碱联合布地奈德混悬液雾化吸入治疗哮喘可有效改善患者的肺功能和临床症状,调节炎性因子水平,且药物副作用少。