Sequence-to-sequence translation from mass spectra to peptides with a transformer model.

A fundamental challenge in mass spectrometry-based proteomics is the identification of the peptide that generated each acquired tandem mass spectrum. Approaches that leverage known peptide sequence databases cannot detect unexpected peptides and can be impractical or impossible to apply in some settings. Thus, the ability to assign peptide sequences to tandem mass spectra without prior information-de novo peptide sequencing-is valuable for tasks including antibody sequencing, immunopeptidomics, and metaproteomics. Although many methods have been developed to address this problem, it remains an outstanding challenge in part due to the difficulty of modeling the irregular data structure of tandem mass spectra. Here, we describe Casanovo, a machine learning model that uses a transformer neural network architecture to translate the sequence of peaks in a tandem mass spectrum into the sequence of amino acids that comprise the generating peptide. We train a Casanovo model from 30 million labeled spectra and demonstrate that the model outperforms several state-of-the-art methods on a cross-species benchmark dataset. We also develop a version of Casanovo that is fine-tuned for non-enzymatic peptides. Finally, we demonstrate that Casanovo's superior performance improves the analysis of immunopeptidomics and metaproteomics experiments and allows us to delve deeper into the dark proteome.

A learned score function improves the power of mass spectrometry database search.

MOTIVATION: One of the core problems in the analysis of protein tandem mass spectrometry data is the peptide assignment problem: determining, for each observed spectrum, the peptide sequence that was responsible for generating the spectrum. Two primary classes of methods are used to solve this problem: database search and de novo peptide sequencing. State-of-the-art methods for de novo sequencing use machine learning methods, whereas most database search engines use hand-designed score functions to evaluate the quality of a match between an observed spectrum and a candidate peptide from the database. We hypothesized that machine learning models for de novo sequencing implicitly learn a score function that captures the relationship between peptides and spectra, and thus may be re-purposed as a score function for database search. Because this score function is trained from massive amounts of mass spectrometry data, it could potentially outperform existing, hand-designed database search tools. RESULTS: To test this hypothesis, we re-engineered Casanovo, which has been shown to provide state-of-the-art de novo sequencing capabilities, to assign scores to given peptide-spectrum pairs. We then evaluated the statistical power of this Casanovo score function, Casanovo-DB, to detect peptides on a benchmark of three mass spectrometry runs from three different species. In addition, we show that re-scoring with the Percolator post-processor benefits Casanovo-DB more than other score functions, further increasing the number of detected peptides.

Biomechanical analysis for different mandibular total distalization methods with clear aligners: A finite element study.

Objective: : The purpose of this finite element method (FEM) study was to analyze the biomechanical differences and tooth displacement patterns according to the traction direction, methods, and sites for total distalization of the mandibular dentition using clear aligner treatment (CAT). Methods: : A finite element analysis was performed on four FEM models using different traction methods (via a precision cut hook or button) and traction sites (mandibular canine or first premolar). A distalization force of 1.5 N was applied to the traction site by changing the direction from -30 to +30° to the occlusal plane. The initial tooth displacement and von Mises stress on the clear aligners were analyzed. Results: : All CAT-based total distalization groups showed an overall trend of clockwise or counterclockwise rotation of the occlusal plane as the force direction varied. Mesiodistal tipping of individual teeth was more prominent than that of bodily movements. The initial displacement pattern of the mandibular teeth was more predominant based on the traction site than on the traction method. The elastic deformation of clear aligners is attributed to unintentional lingual tipping or extrusion of the mandibular anterior teeth. Conclusions: : The initial tooth displacement can vary according to different distalization strategies for CAT-based total distalization. Discreet application and biomechanical understanding of traction sites and directions are necessary for appropriate mandibular total distalization.

Evaluating Proteomics Imputation Methods with Improved Criteria.

Quantitative measurements produced by tandem mass spectrometry proteomics experiments typically contain a large proportion of missing values. Missing values hinder reproducibility, reduce statistical power, and make it difficult to compare across samples or experiments. Although many methods exist for imputing missing values, in practice, the most commonly used methods are among the worst performing. Furthermore, previous benchmarking studies have focused on relatively simple measurements of error such as the mean-squared error between imputed and held-out values. Here we evaluate the performance of commonly used imputation methods using three practical, "downstream-centric" criteria. These criteria measure the ability to identify differentially expressed peptides, generate new quantitative peptides, and improve the peptide lower limit of quantification. Our evaluation comprises several experiment types and acquisition strategies, including data-dependent and data-independent acquisition. We find that imputation does not necessarily improve the ability to identify differentially expressed peptides but that it can identify new quantitative peptides and improve the peptide lower limit of quantification. We find that MissForest is generally the best performing method per our downstream-centric criteria. We also argue that existing imputation methods do not properly account for the variance of peptide quantifications and highlight the need for methods that do.

Effects of different distalization directions and methods on maxillary total distalization with clear aligners: a finite element study.

OBJECTIVES: To analyze the effects of maxillary tooth distalization by clear aligner (CA) treatment with variations in the angular direction of the distalization force, presence of attachments, and force-application method used. MATERIALS AND METHODS: A finite element model containing alveolar bone, dentition, and periodontal ligament was constructed. Analytical model groups were as follows: (1) distalization with buttons (without attachments), (2) buttons on canines (with attachments), (3) precision cuts on the canines (without attachments), and (4) precision cuts on the canines (with attachments). A distalization force of 1.5 N was applied to the button or precision cut at -30°, -20°, -10°, 0°, 10°, 20°, and 30° to the occlusal plane. RESULTS: As the direction of force approached +30°, the dentition inclined posteriorly. The posterior movement pattern was significantly influenced by the presence of an attachment and the direction of force, rather than the area where the force was applied. Distal inclination was dramatically reduced with attachments. A disengagement or deformation of the CA may reduce the distalization efficiency of the dentition or move the dentition in an inappropriate direction. CONCLUSIONS: Attachments for efficient distalization by the CA are necessary. The use of miniscrews in the direction of force parallel to the occlusal plane is more advantageous than the use of Class II elastics. Due to CA deformation, distalization with the button on the canines can be more effective for distal movement of the maxillary dentition.

Comparison of root apex's position between unilateral and bilateral palatally impacted canines: A pilot study.

INTRODUCTION: The purpose was to determine whether the location of the root apices of unilateral palatally impacted canines differs from that of bilateral palatally impacted canines using cone-beam computed tomography. METHODS: The subjects were divided into 3 groups: a bilateral palatally impacted canine group (BPG; n = 28), a unilateral palatally impacted canine group (UPG; n = 28), and a control group (CG; n = 28) that included contralateral normally erupted canines in the UPG. After selecting the root apex in the onDemand3D program, 3-dimensional coordinates were extracted. These 3-dimensional coordinates were converted using the MATLAB program to 2-dimensional coordinates via projection on the palatal plane. Procrustes analysis was used to superimpose these 2-dimensional coordinates. The x- and y-coordinates of the root apices were used to measure the distance between the origin and root apex. RESULTS: The distance between the root apex of the canine and the origin was 17.43 ± 1.78 mm in BPG, 17.96 ± 1.87 mm in UPG, and 13.96 ± 0.95 mm in CG. There was no statistically significant difference between UPG and BPG. However, there was a statistically significant difference between the CG and impacted groups (UPG and BPG). The same results were found for the x- and y-coordinates. CONCLUSIONS: The location of the root apices of unilateral palatally impacted canines is similar to that of bilateral palatally impacted. The location of root apices of palatally impacted canines differs from that of normally erupted canines.

YH29407 with anti-PD-1 ameliorates anti-tumor effects via increased T cell functionality and antigen presenting machinery in the tumor microenvironment.

Among cancer cells, indoleamine 2, 3-dioxygenase1 (IDO1) activity has been implicated in improving the proliferation and growth of cancer cells and suppressing immune cell activity. IDO1 is also responsible for the catabolism of tryptophan to kynurenine. Depletion of tryptophan and an increase in kynurenine exert important immunosuppressive functions by activating regulatory T cells and suppressing CD8+ T and natural killer (NK) cells. In this study, we compared the anti-tumor effects of YH29407, the best-in-class IDO1 inhibitor with improved pharmacodynamics and pharmacokinetics, with first and second-generation IDO1 inhibitors (epacadostat and BMS-986205, respectively). YH29407 treatment alone and anti-PD-1 (aPD-1) combination treatment induced significant tumor suppression compared with competing drugs. In particular, combination treatment showed the best anti-tumor effects, with most tumors reduced and complete responses. Our observations suggest that improved anti-tumor effects were caused by an increase in T cell infiltration and activity after YH29407 treatment. Notably, an immune depletion assay confirmed that YH29407 is closely related to CD8+ T cells. RNA-seq results showed that treatment with YH29407 increased the expression of genes involved in T cell function and antigen presentation in tumors expressing ZAP70, LCK, NFATC2, B2M, and MYD88 genes. Our results suggest that an IDO1 inhibitor, YH29407, has enhanced PK/PD compared to previous IDO1 inhibitors by causing a change in the population of CD8+ T cells including infiltrating T cells into the tumor. Ultimately, YH29407 overcame the limitations of the competing drugs and displayed potential as an immunotherapy strategy in combination with aPD-1.

Reducing Peptide Sequence Bias in Quantitative Mass Spectrometry Data with Machine Learning.

Quantitative mass spectrometry measurements of peptides necessarily incorporate sequence-specific biases that reflect the behavior of the peptide during enzymatic digestion and liquid chromatography and in a mass spectrometer. These sequence-specific effects impair quantification accuracy, yielding peptide quantities that are systematically under- or overestimated. We provide empirical evidence for the existence of such biases, and we use a deep neural network, called Pepper, to automatically identify and reduce these biases. The model generalizes to new proteins and new runs within a related set of tandem mass spectrometry experiments, and the learned coefficients themselves reflect expected physicochemical properties of the corresponding peptide sequences. The resulting adjusted abundance measurements are more correlated with mRNA-based gene expression measurements than the unadjusted measurements. Pepper is suitable for data generated on a variety of mass spectrometry instruments and can be used with labeled or label-free approaches and with data-independent or data-dependent acquisition.

Prediction of maxillary canine impaction using eruption pathway and angular measurement on panoramic radiographs.

OBJECTIVES: To compare the normal eruption pattern and angulation in impacted maxillary canines using panoramic radiographs to predict maxillary canine impaction. MATERIALS AND METHODS: Patients aged 6 to 15 years were classified into the normal eruption group (n = 229) and the impaction group (n = 191). At least two panoramic radiographs were taken in the normal eruption group during the eruption process of the maxillary canine. The growth pattern of the maxillary canine was analyzed using an XY coordinate system, with the tip of the maxillary lateral incisor as the origin and the tooth's long axis as the Y-axis and measurement of the relative position of the crown tip and angulation of the maxillary canine. RESULTS: The crown tips of normally erupted maxillary canines were intensively distributed along the distal surface of the maxillary lateral incisor, while those of impacted canines were widely distributed. The angulations of the normally erupted canines increased as eruption increased along the lateral incisor and then decreased at the cervical point of the lateral incisor. The angulations of the impacted canines were scattered, with no uniform pattern. CONCLUSIONS: While using the normal eruption path of the maxillary canine and the pattern of change in angulation based on the distal surface of the maxillary lateral incisor, early intervention or regular follow-up is needed to prevent maxillary canine impaction.

Cardiac Safety Assessment of Lazertinib: Findings From Patients With EGFR Mutation-Positive Advanced NSCLC and Preclinical Studies.

INTRODUCTION: Lazertinib is a potent, irreversible, brain-penetrant, mutant-selective, and wild-type-sparing third-generation EGFR tyrosine kinase inhibitor (TKI), creating a wide therapeutic index. Cardiovascular adverse events (AEs), including QT prolongation, decreased left ventricular ejection fraction (LVEF), and heart failure, have emerged as potential AEs with certain EGFR TKI therapies. METHODS: Cardiac safety of lazertinib was evaluated in TKI-tolerant adults with EGFR mutation-positive locally advanced or metastatic NSCLC receiving lazertinib (20-320 mg/d). QT intervals corrected with Fridericia's formula (QTcF) prolongation, time-matched concentration-QTcF relationship, change of LVEF, and cardiac failure-associated AEs were evaluated. The clinical findings were supplemented by the following three preclinical studies: an in vitro hERG inhibition assay, an ex vivo isolated perfused rabbit heart study, and an in vivo telemetry-instrumented beagle dog study. RESULTS: Preclinical evaluation revealed little to no physiological effect on the basis of electrocardiogram, electrophysiological, proarrhythmic, and hemodynamic parameters. Clinical evaluation of 181 patients revealed no clinically relevant QTcF prolongation by centralized electrocardiogram in any patient and at any dose level. The predicted magnitude of QTcF value increase at maximum steady-state plasma concentration for the therapeutic dose of lazertinib (240 mg/d) was 2.2 msec (upper bound of the two-sided 90% confidence interval: 3.6 msec). No patient had clinically relevant LVEF decrease (i.e., minimum postbaseline LVEF value of <50% and a maximum decrease in LVEF value from baseline of ≥10 percentage points). Cardiac failure-associated AE occurred in one patient (grade 2 decreased LVEF) and resolved without any dose modifications. CONCLUSIONS: Our first-in-human study, together with preclinical data, indicates that lazertinib is not associated with increased cardiac risk.

Comparison of maxillary basal arch forms using the root apex in adult women with different skeletal patterns: A pilot study.

INTRODUCTION: This study aimed to establish maxillary basal arch forms using the root apices and to determine the differences in the basal arch forms in adult women with different sagittal skeletal patterns. METHODS: This retrospective study included 91 adult women, with either a Class I (n = 24), Class II Division 1 (n = 22), Class II Division 2 (n = 23), or Class III (n = 22) malocclusion, who underwent cone-beam computed tomography. Three-dimensional coordinates of the root apices were determined using the multiplanar reformation mode of OnDemand3D software (Cybermed Inc, Seoul, South Korea). Two-dimensional coordinates were converted from acquired 3-dimensional coordinates via projection on the palatal plane, and the Procrustes superimposition method was used to build the basal arch form. Finally, interroot width measurements were performed for basal arch form comparisons. RESULTS: There were significant differences among the 4 groups (P <0.05) with respect to the intercanine width. The intercanine width of Class II Division 1 group was significantly narrower than that of the other groups. The Class II Division 1 and Class II Division 2 groups tended to have tapered arch forms and squared arch forms, respectively. CONCLUSIONS: We established maxillary basal arch forms using the root apices. The Class II Division 1 group had a significantly narrower intercanine distance. The use of the root apex to depict the basal arch form seems reasonable.

YH25448, an Irreversible EGFR-TKI with Potent Intracranial Activity in EGFR Mutant Non-Small Cell Lung Cancer.

PURPOSE: Given that osimertinib is the only approved third-generation EGFR-TKI against EGFR activating and resistant T790M mutated non-small cell lung cancer (NSCLC), additional mutant-selective inhibitors with a higher efficacy, especially for brain metastases, with favorable toxicity profile are still needed. In this study, we investigated the antitumor efficacy of YH25448, an oral, mutant-selective, irreversible third-generation EGFR-TKI in preclinical models. EXPERIMENTAL DESIGN: Antitumor activity of YH25448 was investigated in vitro using mutant EGFR-expressing Ba/F3 cells and various lung cancer cell lines. In vivo antitumor efficacy, ability to penetrate the blood-brain barrier (BBB), and skin toxicity of YH25448 were examined and compared with those of osimertinib using cell lines and PDX model. RESULTS: Compared with osimertinib, YH25448 showed a higher selectivity and potency in kinase assay and mutant EGFR-expressing Ba/F3 cells. In various cell line models harboring EGFR activating and T790M mutation, YH25448 effectively inhibited EGFR downstream signaling pathways, leading to cellular apoptosis. When compared in vivo at equimolar concentrations, YH25448 produced significantly better tumor regression than osimertinib. Importantly, YH25448 induced profound tumor regression in brain metastasis model with excellent brain/plasma and tumor/brain area under the concentration-time curve value. YH25448 rarely suppressed the levels of p-EGFR in hair follicles, leading to less keratosis than osimertinib in animal model. The potent systemic and intracranial activity of YH25448 has been shown in an ongoing phase I/II clinical trial for advanced EGFR T790M mutated NSCLC (NCT03046992). CONCLUSIONS: Our findings suggest that YH25448 is a promising third-generation EGFR inhibitor, which may be more effective and better tolerated than the currently approved osimertinib.

Notoginseng Radix and Rehmanniae Radix Preparata Extract Combination (YH23537) Reduces Pain and Cartilage Degeneration in Rats with Monosodium Iodoacetate-Induced Osteoarthritis.

Notoginseng Radix and Rehmanniae Radix Preparata have been widely used traditionally for treating inflammatory diseases. This research studies the therapeutic effects of YH23537, the extracts of Notoginseng Radix and Rehmanniae Radix Preparata, on pain and cartilage degeneration in an experimental osteoarthritis (OA) model. Male Wistar rats were inoculated intra-articularly with 3 mg of monosodium iodoacetate (MIA) in the right intra-articular. Four days later, the animals were administrated orally with YH23537 daily for 24 days. Tactile allodynia and weight bearing were measured. Macroscopic and microscopic observations for articular cartilage were performed at the end of the experiment. Protein expression in the joint was determined by immunohistochemistry. The effects of YH23537 on mRNA levels in chondrocytes stimulated with interleukin (IL)-1ß were analyzed using random polymerase chain reaction. OA induction was confirmed by significant decrease of paw withdrawal latency, paw withdrawal threshold, and weight bearing compared with the normal group at 3 days after MIA injection. The YH23537-treated groups displayed significant increases in pain thresholds and weight bearing throughout the observation period. The damage to articular cartilage was significantly lessened visually and histopathologically by YH23537 treatment. YH23537 suppressed the expression of metalloproteinase-3, nitrotyrosine, IL-1ß and IL-6 increased in OA joints. YH23537 upregulated tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-3 in IL-1ß-stimulated human OA chondrocytes. The protein levels of the NF-κBp65 and HIF-2α in the joint tissues were reduced by YH23537. YH23537 exerted antinociceptive effects and cartilage protective effects in experimental OA rats by suppressing oxidative injury, inflammatory mediators, and inducing anabolic factors. We suggest that YH23537 may have efficacy for treating OA in humans.

The novel, potent and highly selective 5-HT4 receptor agonist YH12852 significantly improves both upper and lower gastrointestinal motility.

BACKGROUND AND PURPOSE: 5-HT4 receptor agonists have been shown to be effective at treating various gastrointestinal tract disorders. However, a lack of selectivity against off-targets has been a limiting factor for their clinical use. EXPERIMENTAL APPROACH: The binding affinity and selectivity of YH12852 for human 5-HT4(a) receptor in CHO-K1 cells were evaluated using radioligand binding assays, and agonistic activity was assessed using a ß-lactamase reporter system. Contractile activity and propulsive motility were measured in the guinea pig isolated distal colon. Its prokinetic effect on the gastrointestinal tract was evaluated in guinea pigs, dogs and monkeys. Its tissue distribution was evaluated in rats. KEY RESULTS: YH12852 exhibited high affinity and potency for human recombinant 5-HT4(a) receptor with high selectivity over other 5-HT and non-5-HT receptors, ion channels, enzymes and transporters. YH12852 induced contractions and increased propulsive motility in guinea pig isolated colon. These effects were abolished by the 5-HT4 receptor antagonist GR113808. YH12852 increased defecation more effectively than prucalopride in guinea pigs and dogs and improved gastric emptying more effectively than mosapride in guinea pigs, dogs and monkeys. YH12852 was highly distributed to the gastrointestinal tract as the target organ. CONCLUSION AND IMPLICATIONS: The high in vitro potency and selectivity of YH12852 for 5-HT4 receptor translated into potent in vivo efficacy with good tolerability. YH12852 significantly improved both upper and lower bowel motility in the animal models tested and has the potential to address considerable unmet needs in patients with functional constipation, gastroparesis or both.

How far is the root apex of a unilateral impacted canine from the root apices' arch form?

INTRODUCTION: The purpose of this study was to determine the arch form of the root apices of normally erupting teeth and then determine the differences in the location of the apex of impacted canines relative to normally erupting canines. In addition, we sought to determine whether the labiopalatal position of the impacted canines influences the position of the apices. METHODS: The study included 21 patients with unerupted canines that subsequently had a normal eruption, 21 patients with palatally impacted canines, 27 patients with labially impacted canines, and 17 patients with midalveolus impacted canines. Images were obtained using cone beam computed tomography, and the x, y, and z coordinates of the root apices were determined using Ondemand3D software (Cybermed Co., Seoul, Korea). Two-dimensional coordinates were converted from acquired 3-dimensional coordinates via projection on a palatal plane, and the Procrustes method was used to process the converted 2-dimensional coordinates and to draw the arch forms of the root apices. Finally, we measured the extent of root apex deviation from the arch forms of the root apices. RESULTS: Normally erupting canines showed that even though calcifications may be immature, their positions were aligned with a normal arch form. The root apices of the impacted canines were an average of 6.572 mm away from the root apices' arch form, whereas those of the contralateral nonimpacted canines were an average distance of 2.221 mm away, a statistically significant difference. The palatally impacted canines' root apices distribution tended toward the first premolar root apices. CONCLUSIONS: Incompletely calcified, unerupted teeth with a subsequent normal eruption showed a normal arch form of the root apices. The root apices of impacted canines were farther from the arch forms than were the nonimpacted canines. Also, the root apices of impacted canines in the palatal area showed distributions different from those of the other impacted canine groups.

Association between brain lateralization and mixing ability of chewing side.

BACKGROUND/PURPOSE: Previous studies have suggested that functional dominance in one part of the body can be correlated with functional dominance in another part. Thus, the present research aimed to determine whether brain laterality (handedness, footedness, earedness, and eyedness) was related to mixing ability and chewing side preference. MATERIALS AND METHODS: Fifty-four volunteers who were not undergoing any form of dental treatment took part in this study. Self-defined brain laterality was determined through a questionnaire. The volunteers performed five tasks related to brain laterality, which was identified by the side used to perform three or more of the five tasks. Chewing side preference was determined by observing the main gum location on the occlusal area when volunteers chewed for 30 strokes. Mixing Ability Index (MAI) was measured by analyzing the degree of mixing of two differently colored waxes (height, 3 mm; diameter, 20 mm). Occlusion contact area was measured by taking the maximum intercuspation bite with polysiloxane. RESULTS: Thirty-nine volunteers (72%) showed significant agreement between brain dominance and chewing preference side. The association between brain dominance and MAI was not significant. The occlusal contact area of the dominant side (mean = 48.2 mm2) was significantly wider than that of the nondominant side (25.7 mm2). CONCLUSION: Brain laterality can be explained by the side of functional (preference of the hands, eyes, ears, and feet, and survey) has a positive correlation with chewing preference side. MAI between the brain dominant and nondominant sides was not significant. This shows that mastication efficiency does not differ between dominant and nondominant sides. So, this study suggests that brain dominance is correlated with chewing preference, but it does not affect efficiency of mastication.

Safety and Long-Term Efficacy of Forehead Contouring With a Polycaprolactone-Based Dermal Filler.

BACKGROUND: Recently, a biostimulatory dermal filler based on polycaprolactone (PCL) microspheres was introduced. To our knowledge, no report has examined the safety and efficacy of PCL-based dermal fillers in forehead augmentation. OBJECTIVE: To evaluate the safety and efficacy of forehead augmentation using a PCL-based dermal filler. MATERIALS AND METHODS: The study population consisted of 58 patients (57 women, 98%; 1 man, 2%), aged 20 to 65 years, undergoing forehead augmentation using a PCL-based dermal filler between October 2013 and October 2015 at our clinic. The physicians used the Global Aesthetic Improvement Scale (GAIS) to evaluate its efficacy 1, 3, 6, 12, and 24 months postoperatively. RESULTS: The mean GAIS score at 1, 3, 6, 12, and 24 months was 2.14 ± 0.95, 2.38 ± 0.77, 2.50 ± 0.76, 2.45 ± 0.52, and 2.33 ± 0.50, respectively. The scores increased markedly from 1 to 3 months and were maintained at 24 months. CONCLUSION: A PCL-based dermal filler is a good option for soft tissue augmentation of the forehead, as it is safe and has long-lasting favorable cosmetic efficacy.

Retnla overexpression attenuates allergic inflammation of the airway.

Resistin-like molecule alpha (Retnla), also known as 'Found in inflammatory zone 1', is a secreted protein that has been found in bronchoalveolar lavage (BAL) fluid of ovalbumin (OVA)-induced asthmatic mice and plays a role as a regulator of T helper (Th)2-driven inflammation. However, the role of Retnla in the progress of Th2-driven airway inflammation is not yet clear. To better understand the function of Retnla in Th2-driven airway inflammation, we generated Retnla-overexpressing (Retnla-Tg) mice. Retnla-Tg mice showed increased expression of Retnla protein in BAL fluid and airway epithelial cells. Retnla overexpression itself did not induce any alteration in lung histology or lung function compared to non-Tg controls. However, OVA-sensitized/challenged Retnla-Tg mice had decreased numbers of cells in BAL and inflammatory cells accumulating in the lung. They also showed a reduction in mucus production in the airway epithelium, concomitant with a decreased Muc5ac level. These results were accompanied by reduced levels of Th2 cytokines, including interleukin (IL)-4, IL-5, and IL-13, with no effect on levels of OVA-specific immunoglobulin isotypes. Furthermore, phosphorylation of ERK was markedly reduced in the lungs of OVA-challenged Retnla-Tg mice. Taken together, these results indicates that Retnla protects against Th2-mediated inflammation in an experimental mouse model of asthma, suggesting that therapeutic approaches to enhance the production of Retnla or Retnla-like molecules could be valuable for preventing allergic lung inflammation.

Phytochemicals: a multitargeted approach to gynecologic cancer therapy.

Gynecologic cancers constitute the fourth most common cancer type in women. Treatment outcomes are dictated by a multitude of factors, including stage at diagnosis, tissue type, and overall health of the patient. Current therapeutic options include surgery, radiotherapy, and chemotherapy, although significant unmet medical needs remain in regard to side effects and long-term survival. The efficacy of chemotherapy is influenced by cellular events such as the overexpression of oncogenes and downregulation of tumor suppressors, which together determine apoptotic responses. Phytochemicals are a broad class of natural compounds derived from plants, a number of which exhibit useful bioactive effects toward these pathways. High-throughput screening methods, rational modification, and developments in regulatory policies will accelerate the development of novel therapeutics based on these compounds, which will likely improve overall survival and quality of life for patients.

Identification and characterization of CW108F, a novel ß-carboline compound that promotes cardiomyogenesis of stem cells.

AIMS: The aim of this study was to identify new compounds that induce cardiomyocyte differentiation of stem cells through cell-based screening and investigate lineage specificity and mechanisms in vitro. MAIN METHODS: Embryoid bodies (EBs) formed from TC-1/KH2 mouse embryonic stem cells (ESCs) carrying the gene for enhanced green fluorescent protein (EGFP) under the control of the α-myosin heavy chain (MHC) promoter were treated with test compounds. The number of cardiomyocyte-like (EGFP-expressing) cells in EBs was determined by fluorescence-activated cell sorting. Cardiomyocyte differentiation was further confirmed using lineage-specific biochemical assays and by investigating the expression of cardiomyocyte-specific and "stemness"-associated genes. Nuclear factor-kappaB (NF-κB) signaling activity was measured in A549 cells using a reporter-gene assay. KEY FINDINGS: A ß-carboline compound, designated CW108F, increased the number of mouse ESCs expressing α-MHC promoter-driven EGFP and the proportion of beating EBs. CW108F also increased expression of MHC in P19 stem cells, but did not induce osteogenesis of MC3T3-E1 cells, suggesting lineage-specific activity toward cardiomyocytes. CW108F upregulated expression of cardiac-specific GATA-4 and atrial natriuretic factor (ANF) genes in TC-1/KH2 cells, but downregulated expression of the stemness genes, Oct-4 and brachyury. CW108F inhibited NF-κB transcriptional activity, an effect that might contribute to its cardiomyogenesis-promoting activity. SIGNIFICANCE: The results of this study suggest that the novel ß-carboline, CW108F, promotes the differentiation of ESCs into cardiomyocytes and may be useful for investigating molecular pathways of cardiomyogenesis and generating cardiomyocytes from ESCs.