Discovery of novel anaplastic lymphoma kinase (ALK) and histone deacetylase (HDAC) dual inhibitors exhibiting antiproliferative activity against non-small cell lung cancer.

A series of novel benzimidazole derivatives were designed and synthesised based on the structures of reported oral available ALK inhibitor and HDAC inhibitor, pracinostat. In enzymatic assays, compound 3b, containing a 2-acyliminobenzimidazole moiety and hydroxamic acid side chain, could inhibit both ALK and HDAC6 (IC50 = 16 nM and 1.03 µM, respectively). Compound 3b also inhibited various ALK mutants known to be involved in crizotinib resistance, including mutant L1196M (IC50, 4.9 nM). Moreover, 3b inhibited the proliferation of several cancer cell lines, including ALK-addicted H2228 cells. To evaluate its potential for treating cancers in vivo, 3b was used in a human A549 xenograft model with BALB/c nude mice. At 20 mg/kg, 3b inhibited tumour growth by 85% yet had a negligible effect on mean body weight. These results suggest a attracting route for the further research and optimisation of dual ALK/HDAC inhibitors.

Trajectories of body mass index before the diagnosis of type 2 diabetes in a cohort of Taiwanese adults.

BACKGROUND: Although the prevalence of overweight/obesity is lower in Asian countries, the risk of type 2 diabetes (T2DM) is disproportionally higher. We identified and characterized the trajectory patterns of body mass index (BMI) before the onset of T2DM in a Taiwanese population. METHODS: Using the Taiwan MJ cohort study, we sampled the health examination data of 22,934 participants, including 7618 cases of T2DM and 15,316 controls. We used latent class trajectory analysis to identify distinct groups of pre-disease BMI trajectory. To compare the trajectories of cardiometabolic risk factors among different groups, we used linear mixed-effects models. RESULTS: These 22,934 participants included 13,074 men (57%) and 9860 women (43%) who were on average followed for 9.0 years. We identified three distinct pre-disease BMI trajectories in cases: "stable overweight" (n = 7016, 92.1%), "weight gain" (n = 333, 4.4%) and "obesity" (n = 269, 3.5%). The "stable overweight" group had a mean BMI of 24.6 kg/m2 at 15 years prior to diagnosis, had a 1.2 unit increase during follow-up, and had a mean BMI of 25.8 kg/m2 at the time of diagnosis. The "weight gain" group had the most increasing trends in blood pressure/low-density lipoprotein cholesterol over time. CONCLUSION: The BMI trajectory patterns among individuals who later developed diabetes in Taiwan seemed comparable to that of Western populations, but our population developed T2DM at a much lower BMI. Given that most cases belong to the "stable overweight" group, we also support using a population-based strategy for diabetes prevention instead of focusing on the high risk individuals.

Oral health knowledge, attitudes and behaviours of adults: Findings from a National Oral Health Survey in Singapore.

OBJECTIVES: Prevention complements the curative management of oral diseases. Effective preventive interventions involve the adoption of oral health promoting behaviours. Little is known about the awareness of oral disease and its prevention among Singaporean adults as well as their prevailing oral health attitudes and behaviours. The aim of the study was to describe the oral health knowledge, attitudes and behaviours of adults in Singapore. METHODS: A random sample of adults (≥21 years old) in Singapore was selected to complete an interviewer-administered questionnaire. The questionnaire gathered information about their knowledge of the aetiology, signs and symptoms as well as prevention of dental caries and periodontal disease; attitudes about the value of teeth, locus of control in maintaining oral health and oral health behaviours including toothbrushing, flossing and dental attendance. RESULTS: A total of 1196 adults of weighted mean age 48 years old with almost equal proportions of males and females responded to the questionnaire. Participants were more unaware about the causes of periodontal disease (25.7%) than dental caries (4%). While more than 90% of participants felt that healthy teeth were important and could affect their overall health, many (67.0%) felt it was natural to lose their teeth with old age. Among the participants, 83.5% brushed their teeth twice a day; 41.9% flossed their teeth and 53.9% visited the dentist at least once a year. CONCLUSIONS: The study findings showed good knowledge around dental caries but some gaps around periodontal disease. It also found that participants perceived having limited control over preventing tooth loss. Irregular dental attendance and poor denture wearing habits were also noted. The findings shed light on key areas to focus on through oral health promotion to improve overall oral health.

Estrogen, progesterone, cortisol, brain-derived neurotrophic factor, and vascular endothelial growth factor during the luteal phase of the menstrual cycle in women with premenstrual dysphoric disorder.

The interplay between ovarian hormones, stress, and inflammatory markers in developing premenstrual dysphoric disorder (PMDD) remains inadequately understood. This study investigated the associations of dynamic changes in the levels of estrogen, progesterone, cortisol, brain-derived neurotrophic factor (BDNF), and vascular endothelial growth factor (VEGF) with PMDD during the luteal phase of the menstrual cycle. A total of 58 women with PMDD and 50 healthy women were recruited in this study. These women's estrogen, progesterone, cortisol, BDNF, and VEGF levels were evaluated during the preovulation (PO), mid-luteal (ML), and late-luteal (LL) phases. Furthermore, the severity of P MDD symptoms, depressive symptoms, perceived stress, inattention, craving for sweet foods, and fatigue was assessed. The findings revealed that women with PMDD with higher levels of progesterone during the ML or LL phase or a greater increase (ML-PO) or higher sum (ML + LL) of luteal progesterone level exhibited a greater increase in PMDD symptoms during the luteal phase than did the healthy controls. Furthermore, women with PMDD exhibited higher cortisol levels during the LL phase than did the controls. The BDNF level was negatively correlated with PMDD severity. Furthermore, BDNF and VEGF levels were negatively correlated with inattention and craving for sweet foods among women with PMDD. These results suggest an association between progesterone and the exacerbation of PMDD symptoms during the LL phase. Women with PMDD have relatively high cortisol levels during the LL phase. Future investigations with experimental designs or larger sample sizes are warranted to verify the roles of progesterone and cortisol in the development of PMDD.

Comparative Proteomics Reveals Prolonged Corneal Preservation Impaired Ocular Surface Immunity Accompanied by Fibrosis in Human Stroma.

Cornea transplantation is one of the most commonly performed allotransplantations worldwide. Prolonged storage of donor corneas leads to decreased endothelial cell viability, severe stromal edema, and opacification, significantly compromising the success rate of corneal transplantation. Corneal stroma, which constitutes the majority of the cornea, plays a crucial role in maintaining its shape and transparency. In this study, we conducted proteomic analysis of corneal stroma preserved in Optisol-GS medium at 4 °C for 7 or 14 days to investigate molecular changes during storage. Among 1923 identified proteins, 1634 were quantifiable and 387 were significantly regulated with longer preservation. Compared to stroma preserved for 7 days, proteins involved in ocular surface immunomodulation were largely downregulated while proteins associated with extracellular matrix reorganization and fibrosis were upregulated in those preserved for 14 days. The increase in extracellular matrix structural proteins together with upregulation of growth factor signaling implies the occurrence of stromal fibrosis, which may compromise tissue clarity and cause vision impairments. This study is the first to provide insights into how storage duration affects corneal stroma from a proteomic perspective. Our findings may contribute to future research efforts aimed at developing long-term preservation techniques and improving the quality of preserved corneas, thus maximizing their clinical application.

Association Between Socioeconomic Status and Prevalence of Hypersensitivity Diseases and Autism: A Nationwide Study of Children.

OBJECTIVE: Prior research suggests that children with a low socioeconomic status (SES) background are at an increased risk for special healthcare needs. Conversely, for hypersensitivity-related diseases, many studies reported a lower risk among children with lower SES according to the hygiene hypothesis. We aimed to evaluate the association between SES and several hypersensitivity diseases and autism in a representative American sample. METHODS: We used data from the 2016, 2017 and 2018 US National Survey of Children's Health. A total of 102,341 children aged 0-17 years were included. The dependent variables were doctor-diagnosed allergies, arthritis, asthma, diabetes, and autism. The main SES indicators were family poverty levels, highest education of the reported adults and difficulty in family income. Our analysis used logistic regression that accounted for the survey sampling design. RESULTS: The sample had a mean age of 9.4 ± 5.3 years. The weighted prevalence for allergies was 24.4%, 0.3% for arthritis, 11.9% for asthma, 0.5% for diabetes and 2.6% for autism. Children with adults reporting higher educational levels had greater odds of allergies (adjusted odds ratio and 95% CI: 1.48, 1.23-1.78) than those with lower educational levels. But for all other diseases, most findings suggested that a higher odds of disease was associated with lower SES instead of higher SES. CONCLUSIONS: A low SES background remains an important risk factor for hypersensitivity diseases in children. Most of our results suggested that children with low SES were associated with a higher risk of hypersensitivity diseases and autism.

Structure-Activity Relationship Studies of Antimalarial Plasmodium Proteasome Inhibitors─Part II.

With increasing reports of resistance to artemisinins and artemisinin-combination therapies, targeting the Plasmodium proteasome is a promising strategy for antimalarial development. We recently reported a highly selective Plasmodium falciparum proteasome inhibitor with anti-malarial activity in the humanized mouse model. To balance the permeability of the series of macrocycles with other drug-like properties, we conducted further structure-activity relationship studies on a biphenyl ether-tethered macrocyclic scaffold. Extensive SAR studies around the P1, P3, and P5 groups and peptide backbone identified compound TDI-8414. TDI-8414 showed nanomolar antiparasitic activity, no toxicity to HepG2 cells, high selectivity against the Plasmodium proteasome over the human constitutive proteasome and immunoproteasome, improved solubility and PAMPA permeability, and enhanced metabolic stability in microsomes and plasma of both humans and mice.

Trends in the distribution of body mass index, waist circumference and prevalence of obesity among Taiwanese adults, 1993-2016.

BACKGROUND: Differences in the prevalence of general and abdominal obesity by subgroups such as age, sex, and education have been reported worldwide. Most studies in Taiwan regarding obesity prevalence were targeted at school-aged children or without further stratification by subgroups. Our aim was to examine the age-specific secular trend of body mass index (BMI), waist circumference (WC), and obesity prevalence stratified by sex, education and urbanization levels in Taiwanese adults. METHODS: We used three waves of nationally representative population from the Nutrition and Health Survey in Taiwan (NAHSIT) 1993-1996 (n = 2 989), 2005-2008 (n = 2 495), and 2013-2016 (n = 2 880). The data included standardized measurement of body weight, height, and WC. We conducted a serial cross-sectional analysis among adults aged 20 years or above to examine the age-specific trends of BMI, WC, and the prevalence of underweight, overweight, general obesity, and abdominal obesity with stratification by sex, education, and urbanization levels. RESULTS: The general obesity prevalence was 16%, 21%, and 20% and the abdominal obesity prevalence was 27%, 42%, and 47% in the 1993-1996, 2005-2008, and 2013-2016 surveys, respectively. The age-specific secular trend of BMI differed across subgroups; however, the trend of WC increased rapidly regardless of subgroups, except for women aged ≥60 years. The general obesity prevalence increased noticeably among men, younger- and middle-age adults with high school or higher education, middle- and older-age adults with lower than high school education, people <39 and ≥50 years of age residing in rural areas, and among those between 30 and 59 and ≥70 years of age residing in urban areas. CONCLUSIONS: Although the increasing trend of general obesity prevalence was levelling off among several subgroups, the abdominal obesity prevalence increased significantly and rapidly in Taiwan. Future research in developing effective weight and WC control interventions tailored to different subgroups is urgently needed.

Di-(2-ethylhexyl) Phthalate Promotes Allergic Lung Inflammation by Modulating CD8α+ Dendritic Cell Differentiation via Metabolite MEHP-PPARγ Axis.

Di-(2-ethylhexyl) phthalate (DEHP), a common plasticizer, is a ubiquitous environmental pollutant that can disrupt endocrine function. Epidemiological studies suggest that chronic exposure to DEHP in the environment is associated with the prevalence of childhood allergic diseases; however, the underlying causal relationship and immunological mechanism remain unclear. This study explored the immunomodulatory effect of DEHP on allergic lung inflammation, while particularly focusing on the impact of DEHP and its metabolite on dendritic cell differentiation and activity of peroxisome proliferator-activated receptor gamma (PPARγ). The results showed that exposure to DEHP at a human tolerable daily intake dose exacerbated allergic lung inflammation in mice. Ex vivo flow cytometric analysis revealed that DEHP-exposed mice displayed a significantly decreased number of CD8α+ dendritic cells (DCs) in spleens and DC progenitors in the bone marrow, as well as, less interleukin-12 production in splenic DCs and increased T helper 2 polarization. Pharmacological experiments showed that mono-(2-ethylhexyl) phthalate (MEHP), the main metabolite of DEHP, significantly hampered the differentiation of CD8α+ DCs from Fms-like tyrosine kinase 3 ligand-differentiated bone marrow culture, by modulating PPARγ activity. These results suggested that chronic exposure to DEHP at environmentally relevant levels, promotes allergic lung inflammation, at least in part, by altering DC differentiation through the MEHP-PPARγ axis. This study has crucial implications for the interaction(s) between environmental pollutants and innate immunity, with respect to the development of allergic asthma.

Design, Synthesis, and Optimization of Macrocyclic Peptides as Species-Selective Antimalaria Proteasome Inhibitors.

With over 200 million cases and close to half a million deaths each year, malaria is a threat to global health, particularly in developing countries. Plasmodium falciparum, the parasite that causes the most severe form of the disease, has developed resistance to all antimalarial drugs. Resistance to the first-line antimalarial artemisinin and to artemisinin combination therapies is widespread in Southeast Asia and is emerging in sub-Saharan Africa. The P. falciparum proteasome is an attractive antimalarial target because its inhibition kills the parasite at multiple stages of its life cycle and restores artemisinin sensitivity in parasites that have become resistant through mutation in Kelch K13. Here, we detail our efforts to develop noncovalent, macrocyclic peptide malaria proteasome inhibitors, guided by structural analysis and pharmacokinetic properties, leading to a potent, species-selective, metabolically stable inhibitor.

Benchmarking differential expression, imputation and quantification methods for proteomics data.

Data analysis is a critical part of quantitative proteomics studies in interpreting biological questions. Numerous computational tools for protein quantification, imputation and differential expression (DE) analysis were generated in the past decade and the search for optimal tools is still going on. Moreover, due to the rapid development of RNA sequencing (RNA-seq) technology, a vast number of DE analysis methods were created for that purpose. The applicability of these newly developed RNA-seq-oriented tools to proteomics data remains in doubt. In order to benchmark these analysis methods, a proteomics dataset consisting of proteins derived from humans, yeast and drosophila, in defined ratios, was generated in this study. Based on this dataset, DE analysis tools, including microarray- and RNA-seq-based ones, imputation algorithms and protein quantification methods were compared and benchmarked. Furthermore, applying these approaches to two public datasets showed that RNA-seq-based DE tools achieved higher accuracy (ACC) in identifying DEPs. This study provides useful guidelines for analyzing quantitative proteomics datasets. All the methods used in this study were integrated into the Perseus software, version 2.0.3.0, which is available at https://www.maxquant.org/perseus.

Birth cohort, sex and educational disparities in the trajectories of body mass index in Taiwan: a longitudinal study.

BACKGROUND: Taiwan has gone through rapid industrialization, urbanization and economic growth in the 20th and early 21st centuries. Therefore, the population has experienced significant changes in the physical and social environment during the life course, which may affect the overall adiposity. Our aim was to examine the age trajectories of height, weight and body mass index (BMI) in the Taiwanese population and to explore the influences of sex, birth cohort and education. METHODS: The sample comprised 572,358 residents between 20 and 94 years of age in Taiwan who attended at least one health examination during 1996 to 2017 in a cohort study. Repeated measures of body weight and height were collected using an auto-anthropometer. We conducted a series of linear mixed-effects growth curve models to examine the trajectory of height, weight, and BMI across the life course with stratification by sex. RESULTS: Age-related trajectories of BMI differed between men and women and stronger cohort effects were observed among men, with younger cohorts having higher BMI. After holding cohort and age variables constant, men with junior high or lower education were shorter, thinner and had higher BMI than men with university or higher education (effect sizes: - 3.138 cm, p < 0.001; - 2.277 kg, p < 0.001; 0.121 kg/m2, p < 0.001, respectively). Women with junior high or lower education were shorter, heavier and had higher BMI than women with university or higher education (effect sizes: - 2.368 cm, p < 0.001; 2.417 kg, p < 0.001; 1.691 kg/m2, p < 0.001, respectively). The educational disparities in BMI were found to be larger among women. CONCLUSIONS: Our findings suggest that younger generations, especially men, and lower educational level individuals, particularly women, have increasing levels of BMI. The influence of age and cohort effects together with sex and educational disparities on adiposity should be highlighted when designing future interventions and policies regarding overweight and obesity.

Macrocyclic Peptides that Selectively Inhibit the Mycobacterium tuberculosis Proteasome.

Treatment of tuberculosis (TB) currently takes at least 6 months. Latent Mycobacterium tuberculosis (Mtb) is phenotypically tolerant to most anti-TB drugs. A key hypothesis is that drugs that kill nonreplicating (NR) Mtb may shorten treatment when used in combination with conventional drugs. The Mtb proteasome (Mtb20S) could be such a target because its pharmacological inhibition kills NR Mtb and its genetic deletion renders Mtb unable to persist in mice. Here, we report a series of macrocyclic peptides that potently and selectively target the Mtb20S over human proteasomes, including macrocycle 6. The cocrystal structure of macrocycle 6 with Mtb20S revealed structural bases for the species selectivity. Inhibition of 20S within Mtb by 6 dose dependently led to the accumulation of Pup-tagged GFP that is degradable but resistant to depupylation and death of nonreplicating Mtb under nitrosative stress. These results suggest that compounds of this class have the potential to develop as anti-TB therapeutics.

Oral cancer knowledge and screening behavior among smokers and non-smokers in rural communities.

BACKGROUND: Research suggests having an oral and pharyngeal cancer (OPC) examination for early diagnosis can increase survival rate. However, the OPC screening rate is low in certain populations. To improve OPC screening rate, this study identified factors that are associated with having an OPC examination. METHODS: Participants with landlines and aged 25 years and older were recruited from six northern Florida counties. Bivariate and logistic regressions were used to predict the outcome of whether the participants had ever had an OPC examination as well as whether participants had ever heard of an OPC examination. RESULTS: Of 2260 participants with a mean age of 55.9 ± 15.0 years, the majority of participants never smoked (53.4%), self-identified as Whites (70.6%), and had some college or 2-year degree education (30.3%). Smokers were significantly less likely to have ever heard of an OPC examination than those who never smoked. Significant interaction between smoking status and race, and smoking status and social support interaction were found. Whites who never smoked were more likely to have had an OPC examination than non-Whites who never smoked. Former and current smokers with greater social support were more likely to have had an OPC examination than those with lower social support. CONCLUSION: The findings from this study inform the need to enhance the awareness of having an OPC examination among smokers and to reduce barriers for racial minority populations to receive an OPC examination. Future research is warranted to develop interventions to target certain populations to improve the rate of OPC examination.

Development of a Highly Selective Plasmodium falciparum Proteasome Inhibitor with Anti-malaria Activity in Humanized Mice.

Plasmodium falciparum proteasome (Pf20S) inhibitors are active against Plasmodium at multiple stages-erythrocytic, gametocyte, liver, and gamete activation stages-indicating that selective Pf20S inhibitors possess the potential to be therapeutic, prophylactic, and transmission-blocking antimalarials. Starting from a reported compound, we developed a noncovalent, macrocyclic peptide inhibitor of the malarial proteasome with high species selectivity and improved pharmacokinetic properties. The compound demonstrates specific, time-dependent inhibition of the ß5 subunit of the Pf20S, kills artemisinin-sensitive and artemisinin-resistant P. falciparum isolates in vitro and reduces parasitemia in humanized, P. falciparum-infected mice.

A comparative study on the clinical features of COVID-19 with non-SARS-CoV-2 respiratory viral infections.

During this coronavirus disease 2019 (COVID-19) pandemic, physicians have the important task of risk stratifying patients who present with acute respiratory illnesses. Clinical presentation of COVID-19, however, can be difficult to distinguish from other respiratory viral infections. Thus, identifying clinical features that are strongly associated with COVID-19 in comparison to other respiratory viruses can aid risk stratification and testing prioritization especially in situations where resources for virological testing and resources for isolation facilities are limited. In our retrospective cohort study comparing the clinical presentation of COVID-19 and other respiratory viral infections, we found that anosmia and dysgeusia were symptoms independently associated with COVID-19 and can be important differentiating symptoms in patients presenting with acute respiratory illness. On the other hand, laboratory abnormalities and radiological findings were not statistically different between the two groups. In comparing outcomes, patients with COVID-19 were more likely to need high dependency or intensive care unit care and had a longer median length of stay. With our findings, we emphasize that epidemiological risk factors and clinical symptoms are more useful than laboratory and radiological abnormalities in differentiating COVID-19 from other respiratory viral infections.

Effects of Lower Extremity Muscle Fatigue on Knee Loading During a Forward Drop Jump to a Vertical Jump in Female Athletes.

The aim of this study was to examine changes in the kinematic and kinetic parameters of female athletes performing a forward drop jump to a vertical jump under muscle fatigue condition. Twelve female college athletes performed a forward drop jump to a vertical jump with and without muscle fatigue conditions. A motion capture system and two AMTI force plates were used to synchronously collect kinematic and kinetic data. Inverse dynamics were implemented to calculate the participant's joint loading, joint moment, and energy absorption. A paired sample t-test was used to compare statistical differences between pre-fatigue and post-fatigue conditions (α = .05). The forward trunk lean angle at initial foot contact, as well as the knee range of motion, total negative work and energy absorption contribution of the knee joint during the landing phase were significantly decreased under post-fatigue condition. The increased peak vertical ground reaction force and peak tibial anterior shear forces were also found under post-fatigue condition. These results indicated that muscle fatigue caused participants to change their original landing posture into stiff landing posture and decrease the energy absorption ability, which increased the tibial anterior shear forces. Therefore, female athletes should appropriately increase the knee flexion angle under muscle fatigue condition to reduce the risk of anterior cruciate ligament injuries.

Dendritic cells treated with a prostaglandin I2 analog, iloprost, promote antigen-specific regulatory T cell differentiation in mice.

Iloprost, a stable prostaglandin I2 (PGI2) analog, can inhibit allergic inflammation in an ovalbumin (OVA)-induced asthma model via inhibition of airway dendritic cell (DC) function. However, the underlying mechanism of PGI2 signaling-mediated immunosuppression remains unclear. This study explored whether iloprost-treated DCs can suppress inflammation by promoting antigen-specific regulatory T cell (Treg) differentiation through PGI2-G-protein-coupled receptor (IP). We established an allergic lung inflammation model using a hydrogel biomaterial delivery system and observed that iloprost significantly suppressed OVA-induced Th2 lung inflammation and increased the frequency of OVA-specific Tregs in vivo. We further observed that iloprost-treated DCs displayed tolerogenic characteristics, including low inflammatory cytokine (IL-12, TNF-α, IL-6, IL-23) expression levels, high anti-inflammatory cytokine (IL-10) production, and a semimature phenotype. In addition, iloprost-treated DCs increased OVA-specific CD4+Foxp3+ T cell differentiation from naïve T cells in an IP-dependent pathway in vitro and in vivo. Blocking experiments showed that iloprost-treated DCs promoted Treg differentiation, at least in part, through programmed death ligand 1 (PD-L1), whereas iloprost-induced PD-L1 expression in DCs was through the IP receptor. Furthermore, iloprost treatment suppressed DC-mediated airway inflammation and increased the frequency of OVA-specific Tregs through PD-L1 in vivo. Taken together, these results show that PGI2-IP signaling mediated by iloprost in DCs may lead to immune tolerance, suggesting that the PGI2 analog has the potential to be applied therapeutically for tolerogenic DC immunotherapy in autoimmune diseases or allergic asthma.

Improvement of Asparagine Ethylenediamines as Anti-malarial Plasmodium-Selective Proteasome Inhibitors.

The Plasmodium proteasome (Pf20S) emerged as a target for antimalarials. Pf20S inhibitors are active at multiple stages of the parasite life cycle and synergize with artemisinins, suggesting that Pf20S inhibitors have potential to be prophylactic, therapeutic, and transmission blocking as well as are useful for combination therapy. We recently reported asparagine ethylenediamines (AsnEDAs) as immunoproteasome inhibitors and modified AsnEDAs as selective Pf20S inhibitors. Here, we report further a structure-activity relationship study of AsnEDAs for selective inhibition of Pf20S over human proteasomes. Additionally, we show new mutation that conferred resistance to AsnEDAs and collateral sensitivity to an inhibitor of the Pf20S ß2 subunit, the same as previously identified resistant mutation. This resistance could be overcome through the use of the structure-guided inhibitor design. Collateral sensitivity to inhibitors among respective proteasome subunits underscores the potential value of treating malaria with combinations of inhibitors of different proteasome subunits to minimize the emergence of drug resistance.