Inhaled heparin: Past, present, and future.

While heparin has traditionally served as a key anticoagulant in clinical practice for nearly a century, recent years have witnessed a growing interest in its role as a potent antiinflammatory and antiviral agent, as well as an anticancer agent. To address challenges with injection-based delivery, exploring patient-friendly routes such as oral and pulmonary delivery is crucial. This review specifically highlights the multiple therapeutic benefits of inhaled heparin. In summary, this review serves as a valuable source of information, providing deep insights into the diverse therapeutic advantages of inhaled heparin and its potential applications within clinical contexts.

Dynamic changes of immunocyte subpopulations in thermogenic activation of adipose tissues.

Objectives: The effects of cold exposure on whole-body metabolism in humans have gained increasing attention. Brown or beige adipose tissues are crucial in cold-induced thermogenesis to dissipate energy and thus have the potential to combat metabolic disorders. Despite the immune regulation of thermogenic adipose tissues, the overall changes in vital immune cells during distinct cold periods remain elusive. This study aimed to discuss the overall changes in immune cells under different cold exposure periods and to screen several potential immune cell subpopulations on thermogenic regulation. Methods: Cibersort and mMCP-counter algorithms were employed to analyze immune infiltration in two (brown and beige) thermogenic adipose tissues under distinct cold periods. Changes in some crucial immune cell populations were validated by reanalyzing the single-cell sequencing dataset (GSE207706). Flow cytometry, immunofluorescence, and quantitative real-time PCR assays were performed to detect the proportion or expression changes in mouse immune cells of thermogenic adipose tissues under cold challenge. Results: The proportion of monocytes, naïve, and memory T cells increased, while the proportion of NK cells decreased under cold exposure in brown adipose tissues. Conclusion: Our study revealed dynamic changes in immune cell profiles in thermogenic adipose tissues and identified several novel immune cell subpopulations, which may contribute to thermogenic activation of adipose tissues under cold exposure.

PIPET: predicting relevant subpopulations in single-cell data using phenotypic information from bulk data.

Single-cell RNA sequencing has revealed cellular heterogeneity in complex tissues, notably benefiting research on diseases such as cancer. However, the integration of single-cell data from small samples with extensive clinical features in bulk data remains underexplored. In this study, we introduce PIPET, an algorithmic method for predicting relevant subpopulations in single-cell data based on multivariate phenotypic information from bulk data. PIPET generates feature vectors for each phenotype from differentially expressed genes in bulk data and then identifies relevant cellular subpopulations by assessing the similarity between single-cell data and these vectors. Subsequently, phenotype-related cell states can be analyzed based on these subpopulations. In simulated datasets, PIPET showed robust performance in predicting multiclassification cellular subpopulations. Application of PIPET to lung adenocarcinoma single-cell RNA sequencing data revealed cellular subpopulations with poor survival and associations with TP53 mutations. Similarly, in breast cancer single-cell data, PIPET identified cellular subpopulations associated with the PAM50 clinical subtypes and triple-negative breast cancer subtypes. Overall, PIPET effectively identified relevant cellular subpopulations in single-cell data, guided by phenotypic information from bulk data. This approach comprehensively delineates the molecular characteristics of each cellular subpopulation, offering insights into disease-related subpopulations and guiding personalized treatment strategies.

Gut microbiota-derived gamma-aminobutyric acid from metformin treatment reduces hepatic ischemia/reperfusion injury through inhibiting ferroptosis.

Hepatic ischemia/reperfusion injury (HIRI) is a common and inevitable factor leading to poor prognosis in various liver diseases, making the outcomes of current treatments in clinic unsatisfactory. Metformin has been demonstrated to be beneficial to alleviate HIRI in recent studies, however, the underpinning mechanism remains unclear. In this study, we found metformin mitigates HIRI-induced ferroptosis through reshaped gut microbiota in mice, which was confirmed by the results of fecal microbiota transplantation treatment but showed the elimination of the beneficial effects when gut bacteria were depleted using antibiotics. Detailedly, through 16S rRNA and metagenomic sequencing, we identified that the metformin-reshaped microbiota was characterized by the increase of gamma-aminobutyric acid (GABA) producing bacteria. This increase was further confirmed by the elevation of GABA synthesis key enzymes, glutamic acid decarboxylase and putrescine aminotransferase, in gut microbes of metformin-treated mice and healthy volunteers. Furthermore, the benefit of GABA against HIRI-induced ferroptosis was demonstrated in GABA-treated mice. Collectively, our data indicate that metformin can mitigate HIRI-induced ferroptosis by reshaped gut microbiota, with GABA identified as a key metabolite.

Ternary Dry Powder Agglomerate Inhalation Formulation of Melatonin With Air Jet Mixing to Improve In Vitro And In Vivo Performance.

An improved agglomerate formulation with melatonin and fine lactose for dry powder inhalation using Turbuhaler® was developed. Co-grinding lactose with 1 % magnesium stearate prior to air jet mixing served as a key factor to improve the in vitro aerosolization and in vivo efficacy. Elevated mixing pressure facilitated the dispersion and homogenization of the cohesive mixture for even distribution of agglomerate size after spheroidization and subsequent higher emitted dose with lower variation. Magnesium stearate was employed as a tertiary component to adjust the interparticle force for better aerosolization. At optimized mixing pressure, co-grinding lactose with magnesium stearate before jet mixing displayed further improvement of fine particle fraction to 71.6 ± 3.1 %. The superior fine particle deposition efficiency contributed to rapid onset of action and a high bioavailability of 67.0 % after intratracheal administration to rats. Overall, an inhalable melatonin dry powder formulation exhibiting good aerosol property and lung deposition with clinical translation potential was developed.

CXCL13 promotes thermogenesis in mice via recruitment of M2 macrophage and inhibition of inflammation in brown adipose tissue.

Introduction: Brown adipose tissue (BAT) is mainly responsible for mammalian non-shivering thermogenesis and promotes energy expenditure. Meanwhile, similar to white adipose tissue (WAT), BAT also secretes a variety of adipokines to regulate metabolism through paracrine, autocrine, or endocrine ways. The chemokine C-X-C motif chemokine ligand-13 (CXCL13), a canonical B cell chemokine, functions in inflammation and tumor-related diseases. However, the role of CXCL13 in the adipose tissues is unclear. Methods: The expression of CXCL13 in BAT and subcutaneous white adipose tissue (SWAT) of mice under cold stimulation were detected. Local injection of CXCL13 into BAT of normal-diet and high-fat-diet induced obese mice was used to detect thermogenesis and determine cold tolerance. The brown adipocytes were treated with CXCL13 alone or in the presence of macrophages to determine the effects of CXCL13 on thermogenic and inflammation related genes expression in vitro. Results: In this study, we discovered that the expression of CXCL13 in the stromal cells of brown adipose tissue significantly elevated under cold stimulation. Overexpression of CXCL13 in the BAT via local injection could increase energy expenditure and promote thermogenesis in obese mice. Mechanically, CXCL13 could promote thermogenesis via recruiting M2 macrophages in the BAT and, in the meantime, inhibiting pro-inflammatory factor TNFα level. Discussion: This study revealed the novel role of adipose chemokine CXCL13 in the regulation of BAT activity and thermogenesis.

Supramolecular Transistors with Quantum Interference Effect.

The charge transport through supramolecular junctions exhibits unique quantum interference (QI) effects, which provide an opportunity for the design of supramolecular transistors. Benefiting from the configuration dependence of QI, configuration control of the supramolecular assemblies to demonstrate the QI features is a key but challenging step. In this work, we fabricated the supramolecular transistors and investigated the charge transport through the conducting channel of the individual π-stacked thiophene/phenylene co-oligomers (TPCOs) using the electrochemically gated scanning tunneling microscope break junction technique. We controlled the configuration of the supramolecular channel and switched the QI features between the anti-resonance and resonance states of the supramolecular channels. We observed the supramolecular transistor with its on/off ratio above 103 (∼1300), a high gating efficiency of ∼165 mV/dec, a low off-state leakage current of ∼30 pA, and the channel length scaled down to <2.0 nm. Density functional theory calculations suggested that the QI features in π-stacked TPCOs vary depending on the supramolecular architecture and can be manipulated efficiently by fine-tuning the supramolecular configurations. This work reveals the potential of the supramolecular channels for molecular electronics and provides a fundamental understanding of intermolecular charge transport.

Mechanistic Insight into the Internalization, Distribution, and Autophagy Process of Manganese Nanoparticles in Capsicum annuum L.: Evidence from Orthogonal Microscopic Analysis.

Orthogonal techniques were used to track manganese nanoparticles (MnNPs) in Capsicum annuum L. leaf tissue and cell compartments and subsequently to explain the mechanism of uptake, translocation, and cellular interaction. C. annuum L was cultivated and foliarly exposed to MnNPs (100 mg/L, 50 mL/per leaf) before analysis by using scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy (SEM-EDS) as well as dark-field hyperspectral and two-photon microscopy. We visualized the internalization of MnNP aggregates from the leaf surface and observed particle accumulation in the leaf cuticle and epidermis as well as spongy mesophyll and guard cells. These techniques enabled a description of how MnNPs cross different plant tissues as well as selectively accumulate and translocate in specific cells. We also imaged abundant fluorescent vesicles and vacuoles containing MnNPs, indicating likely induction of autophagy processes in C. annuum L., which is the bio-response upon storing or transforming the particles. These findings highlight the importance of utilizing orthogonal techniques to characterize nanoscale material fate and distribution with complex biological matrices and demonstrate that such an approach offers a significant mechanistic understanding that can inform both risk assessment and efforts aimed at applying nanotechnology to agriculture.

Investigation on the influence of design features on the performance of dry powder inhalers: Spiral channel, mouthpiece dimension, and gas inlet.

As inhaler design is rarely studied but critically important in pulmonary drug delivery, this study investigated the influence of inhaler designs, including a novel spiral channel, mouthpiece dimensions (diameter and length) as well as gas inlet. Experimental dispersion of a carrier-based formulation in conjugation with computational fluid dynamics (CFD) analysis, was performed to determine how the designs affect inhaler performance. Results reveal that inhalers with a narrow spiral channel could effectively increase drug-carrier detachment by introducing high velocity and strong turbulent flow in the mouthpiece, although the drug retention in the device is significantly high. It is also found that reducing mouthpiece diameter and gas inlet size could greatly improve the fine particle dose delivered to the lungs, whereas the mouthpiece length plays a trivial influence on the aerosolization performance. This study contributes toward a better understanding of inhaler designs as relevant to overall inhaler performance, and sheds light on how the designs affect device performance.

Chiroptical Activity of An Achiral Emissive Eu Metal-Organic Framework.

Chiroptical activity of achiral crystals is theoretically allowed but very unusual. There is a particularly scarcity of empirical studies on optically active achiral metal-organic frameworks (MOFs). Herein we report an achiral emissive Eu MOF and its chiroptical properties both in the ground and excited states. The framework crystallizes in an achiral space group (Pna21 ) belonging to the polar point group (mm2), where the asymmetric arrangement of racemic trinuclear Eu-oxo clusters is responsible for the optical activity. A pair of circular dichroisms (CD) and circularly polarized luminescence (CPL) peaks with opposite signs were observed for single crystals. Importantly, the luminescence dissymmetry factor can reach up to 1.1×10-3 , which is comparable in magnitude to the value of most of the chiral-linker-bridged MOFs. This work gives the first example of achiral MOFs with CPL response and should be instructive for the discovery of more CPL emitters from racemic MOF family.

Metabolism-associated molecular classification of gastric adenocarcinoma.

Most gastric cancers (GC) are adenocarcinomas, whereas GC is a highly heterogeneous disease due to its molecular heterogeneity. However, traditional morphology-based classification systems, including the WHO classification and Lauren's classification, have limited utility in guiding clinical treatment. We performed nonnegative matrix factorization (NMF) clustering based on 2752 metabolism-associated genes. We characterized each of the subclasses from multiple angles, including subclass-associated metabolism signatures, immune cell infiltration, clinic10al characteristics, drug sensitivity, and pathway enrichment. As a result, four subtypes were identified: immune suppressed, metabolic, mesenchymal/immune exhausted and hypermutated. The subtypes exhibited significant prognostic differences, which suggests that the metabolism-related classification has clinical significance. Metabolic and hypermutated subtypes have better overall survival, and the hypermutated subtype is likely to be sensitive to anti-PD-1 immunotherapy. In addition, our work showed a strong connection with previously established classifications, especially Lei's subtype, to which we provided an interpretation based on the immune cell infiltration perspective, deepening the understanding of GC heterogeneity. Finally, a 120-gene classifier was generated to determine the GC classification, and a 10-gene prognostic model was developed for survival time prediction.

Identification and validation of an inflammation-related lncRNAs signature for improving outcomes of patients in colorectal cancer.

Background: Colorectal cancer is the fourth most deadly cancer worldwide. Although current treatment regimens have prolonged the survival of patients, the prognosis is still unsatisfactory. Inflammation and lncRNAs are closely related to tumor occurrence and development in CRC. Therefore, it is necessary to establish a new prognostic signature based on inflammation-related lncRNAs to improve the prognosis of patients with CRC. Methods: LASSO-penalized Cox analysis was performed to construct a prognostic signature. Kaplan-Meier curves were used for survival analysis and ROC curves were used to measure the performance of the signature. Functional enrichment analysis was conducted to reveal the biological significance of the signature. The R package "maftool" and GISTIC2.0 algorithm were performed for analysis and visualization of genomic variations. The R package "pRRophetic", CMap analysis and submap analysis were performed to predict response to chemotherapy and immunotherapy. Results: An effective and independent prognostic signature, IRLncSig, was constructed based on sixteen inflammation-related lncRNAs. The IRLncSig was proved to be an independent prognostic indicator in CRC and was superior to clinical variables and the other four published signatures. The nomograms were constructed based on inflammation-related lncRNAs and detected by calibration curves. All samples were classified into two groups according to the median value, and we found frequent mutations of the TP53 gene in the high-risk group. We also found some significantly amplificated regions in the high-risk group, 8q24.3, 20q12, 8q22.3, and 20q13.2, which may regulate the inflammatory activity of cancer cells in CRC. Finally, we identified chemotherapeutic agents for high-risk patients and found that these patients were more likely to respond to immunotherapy, especially anti-CTLA4 therapy. Conclusion: In short, we constructed a new signature based on sixteen inflammation-related lncRNAs to improve the outcomes of patients in CRC. Our findings have proved that the IRLncSig can be used as an effective and independent marker for predicting the survival of patients with CRC.

The effects of grid design on the performance of 3D-printed dry powder inhalers.

The grid structure is an indispensable part of most dry powder inhalers, but the effects of grid geometry on inhaler performance are rarely reported. This study aims to systemically investigate the influence of grid design on the aerosolization performance of capsule-based inhalers through experiments and computational analysis. In-vitro aerosolization and deposition performance of commercial and 3D-printed customized inhalers with different grid mesh designs were experimentally studied using a Next Generation Impactor (NGI). Flow fields in the inhalers were generated, and average turbulence kinetic energy (TKE) and airstream trajectories were obtained through Computational Fluid Dynamics (CFD) analysis, delineating the effects of the different grid designs. Comparative studies using the commercial inhalers and the 3D-printed inhalers show a slightly better performance for the latter, probably due to the different materials used for the inhalers, confirming the suitability of 3D printing. Experimental results show that intensive grid meshes with a relatively small aperture size are beneficial to enhancing inhaler performance. Computational results illustrate that the intensive grid meshes can reduce vortexed airstreams and increase turbulent kinetic energy at the grids in general, which also supports the experimental results. In summary, inhalers with intensive grid meshes are preferred for capsule-based inhalers to enhance aerosolization performance. These findings have significant implications for the comprehensive understanding of how grid designs influence inhaler performance.

Tomato Fruit Nutritional Quality Is Altered by the Foliar Application of Various Metal Oxide Nanomaterials.

Carbohydrates and phytonutrients play important roles in tomato fruit's nutritional quality. In the current study, Fe3O4, MnFe2O4, ZnFe2O4, Zn0.5Mn0.5Fe2O4, Mn3O4, and ZnO nanomaterials (NMs) were synthesized, characterized, and applied at 250 mg/L to tomato plants via foliar application to investigate their effects on the nutritional quality of tomato fruits. The plant growth cycle was conducted for a total of 135 days in a greenhouse and the tomato fruits were harvested as they ripened. The lycopene content was initially reduced at 0 stored days by MnFe2O4, ZnFe2O4, and Zn0.5Mn0.5Fe2O4; however, after a 15-day storage, there was no statistical difference between the treatments and the control. Moreover, the ß-carotene content was also reduced by Zn0.5Mn0.5Fe2O4, Mn3O4, and ZnO. The effects of the Mn3O4 and ZnO carried over and inhibited the ß-carotene after the fruit was stored. However, the total phenolic compounds were increased by ZnFe2O4, Zn0.5Mn0.5Fe2O4, and ZnO after 15 days of storage. Additionally, the sugar content in the fruit was enhanced by 118% and 111% when plants were exposed to Mn3O4 and ZnO, respectively. This study demonstrates both beneficial and detrimental effects of various NMs on tomato fruit quality and highlights the need for caution in such nanoscale applications during crop growth.

A double-edged effect of manganese-doped graphene quantum dots on salt-stressed Capsicum annuum L.

The objective of the current study is to evaluate both the positive and negative effects of manganese-doped graphene quantum dots (GQD-Mn) on Capsicum annuum L. grown under salt stress. GQD-Mn was synthesized, characterized, and foliar-applied (250 mg/L, 120 mg/L, 60 mg/L) to C. annuum L. before and after the flowering stage, during which 100 mM of NaCl solution was introduced into the soil as salt stress. Controls were designed as absolute control (no nanomaterials or salt) and negative control (no nanomaterials only salt). Herein, we report that GQD-Mn offset the reduction of fruit production in salt-stressed C. annuum L. by around 40 %. However, based on a comprehensive analysis of normal alkanes (n-alkane) using gas chromatography-mass spectrometry (GC-MS), we also observed that the leaf epicuticular wax profile was disturbed by GQD-Mn, as the concentration of long-chain n-alkanes was increased. Meanwhile, the content of magnesium (Mg) and zinc (Zn) indicated a potential promoted photosynthesis activity in C. annuum L leaves. We hypothesize that the optical properties of GQD-Mn allow leaves to utilize light more efficiently, thus improving photosynthetic activities in plants to acclimate salt stress. But the increased light usage also induced heat stress on the leaf surfaces, which caused n-alkanes changes. Our results provided a unique perspective on nano-plant interaction that value both beneficial and toxic effects of nanomaterials, especially when evaluating the safety of nano-enabled agriculture in areas facing harsh environmental conditions such as salinity.

Multi-Omics Integrative Analysis of Lung Adenocarcinoma: An in silico Profiling for Precise Medicine.

Lung adenocarcinoma (LUAD) is one of the most common histological subtypes of lung cancer. The aim of this study was to construct consensus clusters based on multi-omics data and multiple algorithms. In order to identify specific molecular characteristics and facilitate the use of precision medicine on patients we used gene expression, DNA methylation, gene mutations, copy number variation data, and clinical data of LUAD patients for clustering. Consensus clusters were obtained using a consensus ensemble of five multi-omics integrative algorithms. Four molecular subtypes were identified. The CS1 and CS2 subtypes had better prognosis. Based on the immune and drug sensitivity predictions, we inferred that CS1 may be less responsive to immunotherapy and less sensitive to chemotherapeutic drugs. The high immune infiltration of CS2 cells may respond well to immunotherapy. Additionally, the CS2 subtype may also respond to EGFR molecular targeted therapy. The CS3 and CS4 subtypes were associated with poor prognosis. These two subtypes had more mutations, especially TP53 ones, as well as higher sensitivity to chemotherapeutics for lung cancer. However, CS3 was enriched in immune-related pathways and may respond to anti-PD1 immunotherapy. In addition, CS1 and CS4 were less sensitive to ferroptosis inhibitors. We performed a comprehensive analysis of the five types of omics data using five clustering algorithms to reveal the molecular characteristics of LUAD patients. These findings provide new insights into LUAD subtypes and potential clinical treatment strategies to guide personalized management and treatment.

Generation of Environmentally Persistent Free Radicals on Metal-Organic Frameworks.

Environmentally persistent free radicals (EPFRs) have been recognized as one of the important emerging contaminants with biological toxicity, environmental persistence, and global mobility. Previous studies have identified the catalytic role of surface metal oxides in EPFRs formation and illustrated the metal-dependence of EPFRs by studying on various metal oxide nanoparticles and single crystals. However, there is still lack of an understanding on the formation of EPFRs from the point of view of metal sites. Various factors (e.g., crystalline phases and surface species) of metal oxides are regarded to contribute to the generation of EPFRs, which present profound difficulties for scientists to tease apart the impact of metal type. Herein, a laboratory investigation, in terms of the acidity and oxidation strength of metal cations, was conducted by selecting metal-variable isostructural metal-organic frameworks as material platforms. Specifically, we evaluated EPFRs generation on MIL-100(M) (M = Al, Cr, Fe) from chlorine-substituted phenol vapor and catechol under thermal conditions. It is found that high Lewis acidity of metal sites is crucial for capturing the above two phenolic precursors, activating the O-H bond and promoting EPFRs formation. Radical species with half-life as long as 70 days were generated on MIL-100 rich in 5-fold coordinated Al3+ sites. The unpaired electron spin density donation was further confirmed by using 27Al solid-state nuclear magnetic resonance spectroscopy. Despite their higher oxidation power than Al3+, the exposed Cr3+ and Fe3+ sites show undetectable catalytic activity for the formation of EPFRs, because of their insufficient Lewis acidity. Our results suggest that the surface species rather than Lewis acid sites may be a major contributor to the formation of EPFRs on metal oxides like Fe2O3.

Association Analysis between Genetic Variants of elovl5a and elovl5b and Poly-Unsaturated Fatty Acids in Common Carp (Cyprinus carpio).

The allo-tetraploid common carp, one widely cultured food fish, is able to produce poly-unsaturated fatty acids (PUFAs). The genetic markers on the PUFA contents for breeding was limited. The polymorphisms in elovl5a and elovl5b, the rate-limiting enzymes in the PUFA biosynthesis, have not been investigated yet. Herein, we identified one coding SNP (cSNP) in elovl5a associated with the content of one PUFA and two cSNPs in elovl5b with the contents of eight PUFAs. The heterozygous genotypes in these three loci were associated with higher contents than the homozygotes. Together with previously identified two associated cSNPs in fads2b, we found the joint effect of these four cSNPs in fads2b and elovl5b on the PUFA contents with the increased explained percentages of PUFA contents. The genotype combinations of more heterozygotes were associated with higher PUFA contents than the other combinations. Using ten genomic selection programs with all cSNPs in fads2b and elovl5b, we obtained the high and positive correlations between the phenotypes and the estimated breeding values of eight PUFAs. These results suggested that elovl5b might be the major gene corresponding to common carp PUFA contents compared with elovl5a. The cSNP combinations in fads2b and elovl5b and the optimal genomic selection program will be used in the future selection breeding to improve the PUFA contents of common carp.

The future of dry powder inhaled therapy: Promising or discouraging for systemic disorders?

Dry powder inhalation therapy has been shown to be an effective method for treating respiratory diseases like asthma, Chronic Obstructive Pulmonary Diseases and Cystic Fibrosis. It has also been widely accepted and used in clinical practices. Such success has led to great interest in inhaled therapy on treating systemic diseases in the past two decades. The current coronavirus (COVID-19) pandemic also has increased such interest and is triggering more potential applications of dry powder inhalation therapy in vaccines and antivirus drugs. Would the inhaled dry powder therapy on systemic disorders be as encouraging as expected? This paper reviews the marketed and in-development dry powder inhaler (DPI) products on the treatment of systemic diseases, their status in clinical trials, as well as the potential for COVID-19 treatment. The advancements and unmet problems on DPI systems are also summarized. With countless attempts behind and more challenges ahead, it is believed that the dry powder inhaled therapy for the treatment of systemic disorders still holds great potential and promise.

M&M: A maximum duration design with the Maxcombo test for a group sequential trial of an immunotherapy with a random delayed treatment effect.

A random delayed treatment effect is expected in a confirmatory clinical trial for an immunotherapy due to the individual heterogeneity of physiological conditions. For this reason, the delay time will be assumed to follow a continuous distribution that is difficult to estimate accurately based on the early-phase data, which hinders the specification of the most powerful weighted log-rank test. Therefore, we propose a simulation-based maximum duration design with a robustly powerful Maxcombo test for a group sequential trial for the immunotherapy with the random delayed treatment effect. The design obtains the group sequential boundaries by a simulation procedure and determines the required maximum sample size using a one-dimensional search in which another simulation procedure is used to calculate empirical power. The simulation researches proved the accuracy of the group sequential boundaries and their robustness against the misspecified maximum sample sizes for large samples and revealed their moderate sensitivity against the misspecified survival distributions under the null hypothesis of no difference. The studies investigated whether the type I error rate would inflate under the "inferior" null hypothesis and evaluated the robustness against different distributions of the delay time in terms of the empirical power among the Maxcombo tests and component weighted log-rank tests.