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The potent antimalarial drug pyronaridine (PND) was tested for its potential as an anticancer drug. After exposing cancerous (17) and non-cancerous (2) cells to PND for 72 hr, PND was found to exhibit consistent and potent cytotoxic activity at low micromolar (μM) concentrations that ranged from 1.6 μM to 9.4 μM. Moreover, PND exerted a significant selective cytotoxicity index (SCI) on five out of seven breast cancer cell lines tested, with favorable values of 2.5 to 4.4, as compared with the non-cancerous breast MCF-10A cell line. By using the same comparison, PND exhibited a significant SCI on three out of four leukemia/lymphoma cell lines with promising values of 3.3 to 3.5. One breast cancer and one leukemia cell line were tested further in order to determine the likely mode of action of PND. PND was found to consistently elicit phosphatidylserine externalization, mitochondrial depolarization, and DNA fragmentation, in both the triple negative MDA-MB-231 breast cancer and HL-60 leukemia cell lines. In addition, PND treatment altered cell cycle progression in both cancer cells. Subsequent DNA mobility-shift assays, UV-Visible spectroscopic titrations, and circular dichroism (CD) experiments revealed that PND intercalates with DNA. The findings presented in this study indicates that PND induces apoptosis and interfered with cell cycle progression of cancer cell lines and these results indicate that this drug has the potential as a repurposed drug for cancer therapy.

This study aims to investigate the development of pitch-matching, rhythmic entrainment, and socioemotional skills in children who received formal music instruction and other non-music based after school programs. Eighty-three children, averaging 6.81 years old at baseline, were enrolled in either a music, sports, or no after-school program and followed over four years. The music program involved formal and systematic instruction in music theory, instrumental technique, and performance. Most control participants had no music education; however, in some instances, participants received minimal music education at school or at church. Musical development was measured using a pitch-matching and drumming-based rhythmic entrainment task. Sharing behavior was measured using a variation of the dictator game, and empathy was assessed using three different assessments: the Index of Empathy for Children and Adolescence (trait empathy), the Reading the Mind in the Eyes Test (theory of mind), and a Fiction Emotion-Matching task (state empathy). Results revealed no time-related associations in pitch-matching ability; however, formal music instruction improved pitch-matching relative to controls. On the contrary, improvements in rhythmic entrainment were best explained by age-related changes rather than music instruction. This study also found limited support for a positive association between formal music instruction and socioemotional skills. That is, individuals with formal music instruction exhibited improved emotion-matching relative to those with sports training. In terms of general socioemotional development, children's trait-level affective empathy did not improve over time, while sharing, theory of mind, and state empathy did. Additionally, pitch-matching and rhythmic entrainment did not reliably predict any socioemotional measures, with associations being trivial to small. While formal music instruction benefitted pitch-matching ability and emotion-matching to an audiovisual stimulus, it was not a significant predictor of rhythmic entrainment or broader socioemotional development. These findings suggest that the transfer of music training may be most evident in near or similar domains., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Villanueva et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Humans need meaningful social interactions, but little is known about the consequences of not having them. We examined meaningful social interactions and the lack thereof in patients diagnosed with major depressive disorder (MDD) or social phobia (SP) and compared them to a control group (CG). Using event-sampling methodology, we sampled participants' everyday social behavior 6 times per day for 1 week in participants' natural environment. We investigated the quality and the proportion of meaningful social interactions (when they had meaningful social interactions) and degree of wishing for and avoidance of meaningful social interactions (when they did not have meaningful social interactions). Groups differed on the quality and avoidance of meaningful social interactions: Participants with MDD and SP reported perceiving their meaningful social interactions as lower quality (in terms of subjective meaningfulness) than the CG, with SP patients reporting even lower quality than the MDD patients. Further, both MDD and SP patients reported avoiding meaningful social interactions significantly more often than the CG. Although the proportion of meaningful social interactions was similar in all groups, the subjective quality of meaningful social interactions was perceived to be lower in MDD and SP patients. Future research might further identify what variables influenced the reinforcement of the MDD and SP patients so that they engaged in the same number of meaningful social interactions even though the quality of their meaningful social interactions was lower. Increasing awareness of what happens when patients do or do not have meaningful social interactions will help elucidate a potentially exacerbating or maintaining factor of the disorders., Competing Interests: The authors have declared that no competing interests exist.

Background: Contact tracing is a crucial part of the public health surveillance toolkit. However, it is labor-intensive and costly to carry it out. Some countries have faced challenges implementing contact tracing, and no impact evaluations using empirical data have assessed its impact on COVID-19 mortality. This study assesses the impact of contact tracing in a middle-income country, providing data to support the expansion and optimization of contact tracing strategies to improve infection control., Methods: We obtained publicly available data on all confirmed COVID-19 cases in Colombia between March 2 and June 16, 2020. (N = 54,931 cases over 135 days of observation). As suggested by WHO guidelines, we proxied contact tracing performance as the proportion of cases identified through contact tracing out of all cases identified. We calculated the daily proportion of cases identified through contact tracing across 37 geographical units (32 departments and five districts). Further, we used a sequential log-log fixed-effects model to estimate the 21-days, 28-days, 42-days, and 56-days lagged impact of the proportion of cases identified through contact tracing on daily COVID-19 mortality. Both the proportion of cases identified through contact tracing and the daily number of COVID-19 deaths are smoothed using 7-day moving averages. Models control for the prevalence of active cases, second-degree polynomials, and mobility indices. Robustness checks to include supply-side variables were performed., Results: We found that a 10 percent increase in the proportion of cases identified through contact tracing is related to COVID-19 mortality reductions between 0.8% and 3.4%. Our models explain between 47%-70% of the variance in mortality. Results are robust to changes of specification and inclusion of supply-side variables., Conclusion: Contact tracing is instrumental in containing infectious diseases. Its prioritization as a surveillance strategy will substantially impact reducing deaths while minimizing the impact on the fragile economic systems of lower and middle-income countries. This study provides lessons for other LMIC., Competing Interests: The authors have declared that no competing interests exist.

Evidence-based practice (EBP) in management is still in its infancy. Several studies suggest that managers in businesses and other organizations do not consult the scientific evidence when making decisions. To facilitate its uptake, we need to better understand practitioner attitudes and perceived barriers related to EBP. In medicine and nursing, an abundance of research exists on this subject, although such studies are rare in management. To address this gap, we surveyed 2,789 management practitioners in Belgium, the Netherlands, the United States, the United Kingdom and Australia. Our findings indicate that most managers we studied have positive attitudes towards EBP. However, lack of time and a limited understanding of scientific research are perceived as major barriers to the uptake and implementation of EBP in management. Studies in other professions where EBP is far more established also report similar barriers. We discuss the implications of our findings for practice, education and research, providing suggestions to enhance use of EBP in management practice.

A siphon is a device that is used to drain a container, with water rising inside a hose in the form of an inverted U and then going down towards a discharge point placed below the initial water level. The siphon is the first of a number of inventions of the ancients documented about 2.000 years ago by Hero of Alexandria in his treatise Pneumatics, and although the explanation given by Hero was essentially correct, there is nowadays a controversy about the underlying mechanism that explains the working of this device. Discussions concerning the physics of a siphon usually refer to concepts like absolute negative pressures, the strength of liquid's cohesion and the possibility of a siphon working in vacuum or in the presence of bubbles. Torricelli understood the working principle of the barometer and the impossibility of pumping water out of wells deeper than 10.33 m. Following Torricelli's ideas it would also not be possible to build a siphon that drives pure water to ascend higher than 10.33 m. In this work, we report the first siphon that drives water (with surfactant) to ascend higher than the Torricellian limit. Motivated by the rising of sap in trees, we built a 15.4 m siphon that shows that absolute negative pressures are not prohibited, that cohesion plays an important role in transmitting forces through a fluid, and that surfactants can help to the transport of water in a metastable regime of negative pressures.

Background: Avian influenza A (H7N9) virus has emerged recently and continues to cause severe disease with a high mortality rate in humans prompting the development of candidate vaccine viruses. Live attenuated influenza vaccines (LAIV) are 6:2 reassortant viruses containing the HA and NA gene segments from wild type influenza viruses to induce protective immune responses and the six internal genes from Master Donor Viruses (MDV) to provide temperature sensitive, cold-adapted and attenuated phenotypes., Methodology/principal Findings: LAIV candidate A/Anhui/1/2013(H7N9)-CDC-LV7A (abbreviated as CDC-LV7A), based on the Russian MDV, A/Leningrad/134/17/57 (H2N2), was generated by classical reassortment in eggs and retained MDV temperature-sensitive and cold-adapted phenotypes. CDC-LV7A had two amino acid substitutions N123D and N149D (H7 numbering) in HA and one substitution T10I in NA. To evaluate the role of these mutations on the replication capacity of the reassortants in eggs, the recombinant viruses A(H7N9)RG-LV1 and A(H7N9)RG-LV2 were generated by reverse genetics. These changes did not alter virus antigenicity as ferret antiserum to CDC-LV7A vaccine candidate inhibited hemagglutination by homologous A(H7N9) virus efficiently. Safety studies in ferrets confirmed that CDC-LV7A was attenuated compared to wild-type A/Anhui/1/2013. In addition, the genetic stability of this vaccine candidate was examined in eggs and ferrets by monitoring sequence changes acquired during virus replication in the two host models. No changes in the viral genome were detected after five passages in eggs. However, after ten passages additional mutations were detected in the HA gene. The vaccine candidate was shown to be stable in the ferret model; post-vaccination sequence data analysis showed no changes in viruses collected in nasal washes present at day 5 or day 7., Conclusions/significance: Our data indicate that the A/Anhui/1/2013(H7N9)-CDC-LV7A reassortant virus is a safe and genetically stable candidate vaccine virus that is now available for distribution by WHO to vaccine manufacturers.

Highly pathogenic avian influenza (HPAI) H5N1 is endemic in Vietnamese poultry and has caused sporadic human infection in Vietnam since 2003. Human infections with HPAI H5N1 are of concern due to a high mortality rate and the potential for the emergence of pandemic viruses with sustained human-to-human transmission. Viruses isolated from humans in southern Vietnam have been classified as clade 1 with a single genome constellation (VN3) since their earliest detection in 2003. This is consistent with detection of this clade/genotype in poultry viruses endemic to the Mekong River Delta and surrounding regions. Comparison of H5N1 viruses detected in humans from southern Vietnamese provinces during 2012 and 2013 revealed the emergence of a 2013 reassortant virus with clade 1.1.2 hemagglutinin (HA) and neuraminidase (NA) surface protein genes but internal genes derived from clade 2.3.2.1a viruses (A/Hubei/1/2010-like; VN12). Closer analysis revealed mutations in multiple genes of this novel genotype (referred to as VN49) previously associated with increased virulence in animal models and other markers of adaptation to mammalian hosts. Despite the changes identified between the 2012 and 2013 genotypes analyzed, their virulence in a ferret model was similar. Antigenically, the 2013 viruses were less cross-reactive with ferret antiserum produced to the clade 1 progenitor virus, A/Vietnam/1203/2004, but reacted with antiserum produced against a new clade 1.1.2 WHO candidate vaccine virus (A/Cambodia/W0526301/2012) with comparable hemagglutination inhibition titers as the homologous antigen. Together, these results indicate changes to both surface and internal protein genes of H5N1 viruses circulating in southern Vietnam compared to 2012 and earlier viruses.

One of the important lessons learned from the 2009 H1N1 pandemic is that a high yield influenza vaccine virus is essential for efficient and timely production of pandemic vaccines in eggs. The current seasonal and pre-pandemic vaccine viruses are generated either by classical reassortment or reverse genetics. Both approaches utilize a high growth virus, generally A/Puerto Rico/8/1934 (PR8), as the donor of all or most of the internal genes, and the wild type virus recommended for inclusion in the vaccine to contribute the hemagglutinin (HA) and neuraminidase (NA) genes encoding the surface glycoproteins. As a result of extensive adaptation through sequential egg passaging, PR8 viruses with different gene sequences and high growth properties have been selected at different laboratories in past decades. The effect of these related but distinct internal PR8 genes on the growth of vaccine viruses in eggs has not been examined previously. Here, we use reverse genetics to analyze systematically the growth and HA antigen yield of reassortant viruses with 3 different PR8 backbones. A panel of 9 different HA/NA gene pairs in combination with each of the 3 different lineages of PR8 internal genes (27 reassortant viruses) was generated to evaluate their performance. Virus and HA yield assays showed that the PR8 internal genes influence HA yields in most subtypes. Although no single PR8 internal gene set outperformed the others in all candidate vaccine viruses, a combination of specific PR8 backbone with individual HA/NA pairs demonstrated improved HA yield and consequently the speed of vaccine production. These findings may be important both for production of seasonal vaccines and for a rapid global vaccine response during a pandemic.

Lipid molecules such as arachidonic acid (AA) and sphingolipid metabolites have been implicated in modulation of neuronal and endocrine secretion. Here we compare the effects of these lipids on secretion from cultured bovine chromaffin cells. First, we demonstrate that exogenous sphingosine and AA interact with the secretory apparatus as confirmed by FRET experiments. Examination of plasma membrane SNARE microdomains and chromaffin granule dynamics using total internal reflection fluorescent microscopy (TIRFM) suggests that sphingosine production promotes granule tethering while arachidonic acid promotes full docking. Our analysis of single granule release kinetics by amperometry demonstrated that both sphingomyelinase and AA treatments enhanced drastically the amount of catecholamines released per individual event by either altering the onset phase of or by prolonging the off phase of single granule catecholamine release kinetics. Together these results demonstrate that the kinetics and extent of the exocytotic fusion pore formation can be modulated by specific signalling lipids through related functional mechanisms.

Although cyclin dependent kinase (CDK)-2 is known to be dispensable for the growth of most tumors, it is thought to be important for the proliferation of melanoma cells, where its expression is controlled by the melanocyte-lineage specific transcription factor MITF. Treatment of a panel of melanoma cells with the CDK inhibitor dinaciclib led to a concentration-dependent inhibition of growth under both 2D adherent and 3D organotypic cell culture conditions. Dinaciclib targeted melanoma cell lines regardless of cdk2 or MITF levels. Inhibition of growth was associated with a rapid induction of G2/M cell arrest and apoptosis. Treatment of human melanoma mouse xenografts with dinaciclib led to tumor regression associated with reduced retinoblastoma protein phosphorylation and Bcl-2 expression. Further mechanistic studies revealed that dinaciclib induces p53 expression whilst simultaneously downregulating the expression of the anti-apoptotic factors Mcl-1 and XIAP. To clarify the role of p53 activation in the dinaciclib-induced cell death, we generated melanoma cell lines in which p53 expression was knocked down using a shRNA lentiviral vector. Knockdown of p53 completely abolished the induction of apoptosis seen following dinaciclib treatment as shown by a lack of annexin-V staining and caspase-3 cleavage. Altogether, these data show that dinaciclib induces apoptosis in a large panel of melanoma cell lines through a mechanism requiring p53 expression.

Aims: This proof-of-concept study was designed to determine if functional, pharmacodynamic profiles relevant to targeted therapy could be derived from live human melanoma samples using a novel automated platform., Methods: A series of 13 melanoma cell lines was briefly exposed to a BRAF inhibitor (PLX-4720) on a platform employing automated fluidics for sample processing. Levels of the phosphoprotein p-ERK in the mitogen-activated protein kinase (MAPK) pathway from treated and untreated sample aliquots were determined using a bead-based immunoassay. Comparison of these levels provided a determination of the pharmacodynamic effect of the drug on the MAPK pathway. A similar ex vivo analysis was performed on fine needle aspiration (FNA) biopsy samples from four murine xenograft models of metastatic melanoma, as well as 12 FNA samples from patients with metastatic melanoma., Results: Melanoma cell lines with known sensitivity to BRAF inhibitors displayed marked suppression of the MAPK pathway in this system, while most BRAF inhibitor-resistant cell lines showed intact MAPK pathway activity despite exposure to a BRAF inhibitor (PLX-4720). FNA samples from melanoma xenografts showed comparable ex vivo MAPK activity as their respective cell lines in this system. FNA samples from patients with metastatic melanoma successfully yielded three categories of functional profiles including: MAPK pathway suppression; MAPK pathway reactivation; MAPK pathway stimulation. These profiles correlated with the anticipated MAPK activity, based on the known BRAF mutation status, as well as observed clinical responses to BRAF inhibitor therapy., Conclusion: Pharmacodynamic information regarding the ex vivo effect of BRAF inhibitors on the MAPK pathway in live human melanoma samples can be reproducibly determined using a novel automated platform. Such information may be useful in preclinical and clinical drug development, as well as predicting response to targeted therapy in individual patients.