Objective To evaluate response to esketamine nasal spray plus an oral antidepressant (ESK + AD) at day 28 in patients with major depressive disorder (DSM-5) and treatment-resistant depression (TRD) who did not meet response criteria within the first week of treatment.Methods The current study is a pooled post hoc analysis of two phase 3, double-blind, active-controlled studies, conducted between August 2015 and February 2018, comparing ESK + AD with an oral antidepressant plus placebo (AD + PBO). Early treatment response was defined as a ≥ 50% decrease in Montgomery-Åsberg Depression Rating Scale total score at day 2 or days 2 and 8. Response rates at day 28 were determined among those not meeting early response criteria.Results 518 patients in the analysis had day 28 observations (ESK + AD, n = 310; AD + PBO, n = 208). A greater percentage of patients treated with ESK + AD versus AD + PBO met response criteria beginning at day 2 (17.3% [55/318] vs 9.4% [19/203]) and at all subsequent timepoints, including day 28 (58.7% [182/310] vs 45.2% [94/208]). In day 2 nonresponders, 54.9% vs 44.3% (ESK + AD vs AD + PBO, respectively) achieved response at day 28 (P  less then  .01). Similarly, among day 2 and 8 nonresponders, 52.1% vs 42.4% achieved response by day 28 (P = .01). In nonresponders at day 2 and at days 2 and 8, the odds ratio for a response at day 28 was 1.61 (95% CI, 1.09-2.40) with ESK + AD versus 1.56 (95% CI, 1.04-2.35) with AD + PBO.Conclusions Patients with TRD without a demonstrated response within the first week of treatment may still derive benefit from a full 4-week induction course of esketamine nasal spray.Trial Registration ClinicalTrials.gov identifiers NCT02417064 and NCT02418585.New genetic engineering techniques have advanced the field of plant molecular biology, and Agrobacterium-mediated transformation has enabled the discovery of numerous molecular and genetic functions. It has been widely used in many plants, including the economically important crop, soybean. Large-scale genetic analyses are needed to comprehend the molecular mechanisms that underlie the agronomic traits of soybean, and the generation of stable transgenic plants involves a lengthy and laborious process. Agrobacterium rhizogenes-mediated hairy root transformation is a quick and efficient method for investigations of root-specific processes and interactions. Generation of composite plants with transgenic roots and wild-type shoots allows for the study of the genetic mechanisms involved in root biology, such as the Bradyrhizobium-soybean interaction. Here, we provide an updated protocol for generating hairy soybean roots in as little as 18 days in a cost- and space-effective manner and demonstrate possible uses of composite plants with soybean nodulation assays and gene expression analysis as examples. © 2021 Wiley Periodicals LLC. Basic Protocol 1 Soybean hairy root transformation Basic Protocol 2 Soybean nodulation assay Alternate Protocol Soybean nodulation assay in germination pouches Support Protocol Bradyrhizobium japonicum culture preparation for inoculation Basic Protocol 3 Histochemical GUS staining for promoter analysis. A series of repeated questionnaire surveys among 8- to9-year-old school children in the city of Patras, Greece, demonstrated a continuous rise in the prevalence of wheeze/asthma from 1978 to 2003, with a plateau between 2003 and 2008. We further investigated wheeze/asthma trends within the same environment over the last decade. Two follow-up surveys were conducted in 2013 (N = 2554) and 2018 (N = 2648). Physician-diagnosed wheeze and asthma were analyzed in relation to their occurrence (recent-onset within the last 2 years; noncurrent before 2 years; persistent both prior and within the last 2 years). In 2018, spirometry was also performed in participants reporting symptoms and in a sample of healthy controls. The prevalence of current wheeze/asthma declined from 6.9% in 2008% to 5.2% in 2013% and 4.3% in 2018. The persistent and noncurrent wheeze/asthma groups followed this overall trend (P-for-trend <0.001), while the prevalence of recent-onset wheeze/asthma remained unchanged (P-for-trend >0.05). see more Persistent and noncurrent wheezers were also more frequently diagnosed with asthma, in contrast to those with recent-onset wheeze. The FEV z-score was less than -1 in 32.1% of children with recent-onset and in 22.4% of those with persistent wheeze/asthma; both rates were higher than those of the Noncurrent wheeze/asthma group (7.1%; p < .05) and of healthy controls (3.5%; p < .001). The prevalence of childhood wheeze/asthma has declined significantly during the last decade in Greece. The reversing trend may in part be attributed to changing asthma perceptions among physicians and/or parents, especially in the case of younger children with troublesome respiratory symptoms.The prevalence of childhood wheeze/asthma has declined significantly during the last decade in Greece. The reversing trend may in part be attributed to changing asthma perceptions among physicians and/or parents, especially in the case of younger children with troublesome respiratory symptoms.Deletion of the pyruvate kinase muscle (PKM) gene, which is involved in conversion of phosphoenolpyruvate to pyruvate, has been shown to curb lactogenic behavior in Chinese hamster ovary (CHO) cells. This study describes the generation of pyruvate kinase muscle isoforms 1 and 2 knockout (PKM-KO) and pyruvate kinase muscle isoform-1 knockout (PKM1-KO) CHO host cells to understand metabolic shifts that reduce lactate secretion in these cells. Glucose and amino acids uptake levels in wild-type (WT), PKM-KO, and PKM1-KO stable cell lines, expressing two different antibodies, were analyzed in 14-day fed-batch production assays using different vessels. PKM-KO and PKM1-KO cells consumed more glucose per cell, altered amino acids metabolism, had higher flux of pyruvate into the tricarboxylic acid (TCA) cycle, and as previously shown reduced lactate secretion levels compared with the WT cells. Additionally, both PKM-KO and PKM1-KO cells had higher specific productivity and lower cell growth rates compared with the WT cells.