MyD88-dependent signaling drives toll-like receptor-induced trained immunity in macrophages.

Immunocompromised populations are highly vulnerable to developing life-threatening infections. Strategies to protect patients with weak immune responses are urgently needed. Employing trained immunity, whereby innate leukocytes undergo reprogramming upon exposure to a microbial product and respond more robustly to subsequent infection, is a promising approach. Previously, we demonstrated that the TLR4 agonist monophosphoryl lipid A (MPLA) induces trained immunity and confers broad resistance to infection. TLR4 signals through both MyD88- and TRIF-dependent cascades, but the relative contribution of each pathway to induction of trained immunity is unknown. Here, we show that MPLA-induced resistance to Staphylococcus aureus infection is lost in MyD88-KO, but not TRIF-KO, mice. The MyD88-activating agonist CpG (TLR9 agonist), but not TRIF-activating Poly I:C (TLR3 agonist), protects against infection in a macrophage-dependent manner. MPLA- and CpG-induced augmentation of macrophage metabolism and antimicrobial functions is blunted in MyD88-, but not TRIF-KO, macrophages. Augmentation of antimicrobial functions occurs in parallel to metabolic reprogramming and is dependent, in part, on mTOR activation. Splenic macrophages from CpG-treated mice confirmed that TLR/MyD88-induced reprogramming occurs in vivo. TLR/MyD88-triggered metabolic and functional reprogramming was reproduced in human monocyte-derived macrophages. These data show that MyD88-dependent signaling is critical in TLR-mediated trained immunity.

Does the Establishment of National New Areas Improve Urban Ecological Efficiency? Empirical Evidence Based on Staggered DID Model.

The environmental effects of national new areas have been an important topic but received little attention in academia. This study conducts a quasi-natural experiment using panel data of China's 282 prefecture-level cities from 2006 to 2019, and evaluates the establishment of national new areas on urban ecological efficiency using the staggered difference-in-difference (DID) method, tests the robustness, and further examines the influential mechanism and urban heterogeneity of the empirical results. The results show that the establishment of national new areas has significantly improved urban eco-efficiency. Moreover, the mechanism analysis of the influences shows that national new areas improve urban eco-efficiency by improving urbanization level and urban transportation infrastructure. In addition, the heterogeneity analysis of cities shows that national new areas of cities in eastern and central regions are both significantly improving urban eco-efficiency, while those in western and northeastern regions are not. Furthermore, the promotion effect in the regions of "one new area in one city" model is better than that in "one new area in two cities" model; national new areas in non-resource-based cities show more positive effects on promoting urban eco-efficiency than those in resource-based cities. The conclusions reliably evaluate the results of the current construction of national new areas and provide feasible suggestions for further implementation of the related policy to balance economic development and environmental protection.

Determining dependency and redundancy for identifying gene-gene interaction associated with complex disease.

As interactions among genetic variants in different genes can be an important factor for predicting complex diseases, many computational methods have been proposed to detect if a particular set of genes has interaction with a particular complex disease. However, even though many such methods have been shown to be useful, they can be made more effective if the properties of gene-gene interactions can be better understood. Towards this goal, we have attempted to uncover patterns in gene-gene interactions and the patterns reveal an interesting property that can be reflected in an inequality that describes the relationship between two genotype variables and a disease-status variable. We show, in this paper, that this inequality can be generalized to [Formula: see text] genotype variables. Based on this inequality, we establish a conditional independence and redundancy (CIR)-based definition of gene-gene interaction and the concept of an interaction group. From these new definitions, a novel measure of gene-gene interaction is then derived. We discuss the properties of these concepts and explain how they can be used in a novel algorithm to detect high-order gene-gene interactions. Experimental results using both simulated and real datasets show that the proposed method can be very promising.

Research Status and Molecular Mechanism of the Traditional Chinese Medicine and Antitumor Therapy Combined Strategy Based on Tumor Microenvironment.

Treatment of malignant tumors encompasses multidisciplinary comprehensive diagnosis and treatment and reasonable combination and arrangement of multidisciplinary treatment, which is not a simple superimposition of multiple treatment methods, but a comprehensive consideration of the characteristics and specific conditions of the patients and the tumor. The mechanism of tumor elimination by restoring the body's immune ability is consistent with the concept of "nourishing positive accumulation and eliminating cancer by itself" in traditional Chinese medicine (TCM). The formation and dynamic changes in the tumor microenvironment (TME) involve many different types of cells and multiple signaling pathways. Those changes are similar to the multitarget and bidirectional regulation of immunity by TCM. Discussing the relationship and mutual influence of TCM and antitumor therapy on the TME is a current research hotspot. TCM has been applied in the treatment of more than 70% of cancer patients in China. Data have shown that TCM can significantly enhance the sensitivity to chemotherapeutic drugs, enhance tumor-suppressing effects, and significantly improve cancer-related fatigue, bone marrow suppression, and other adverse reactions. TCM treatments include the application of Chinese medicine monomers, extracts, classic traditional compound prescriptions, listed compound drugs, self-made compound prescriptions, as well as acupuncture and moxibustion. Studies have shown that the TCM functional mechanism related to the positive regulation of cytotoxic T cells, natural killer cells, dendritic cells, and interleukin-12, while negatively regulating of regulatory T cells, tumor-associated macrophages, myeloid-derived suppressive cells, PD-1/PD-L1, and other immune regulatory factors. However, the application of TCM in cancer therapy needs further study and confirmation. This article summarizes the existing research on the molecular mechanism of TCM regulation of the TME and provides a theoretical basis for further screening of the predominant population. Moreover, it predicts the effects of the combination of TCM and antitumor therapy and proposes further developments in clinical practice to optimize the combined strategy.

miR-922 regulates apoptosis, migration, and invasion by targeting SOCS1 in gastric cancer.

Gastric cancer (GC) is the second leading cause of cancer-related death worldwide. Studies have shown that miR-922 facilitates the development of various diseases and tumors. However, the role of miR-922 in GC and related molecular mechanisms are still unrevealed. Current study indicated that miR-922 was overexpressed in GC tissues and cells. The survival rate of patients in high miR-922 expression group is significantly lower than that in low miR-922 expression group. In addition, overexpression of miR-922 observably restrained the apoptosis of SGC7901 cells and promoted SGC7901 cell proliferation, migration, and invasion. TargetScan predicted that suppressors of cytokine signaling 1 (SOCS1) was a potential target of miR-922. miR-922 upregulation profoundly inhibited the expression of SOCS1. Furthermore, the mRNA level of SOCS1 in GC tissues was significantly lower than that in adjacent tissues, indicating that miR-922 promoted the proliferation, invasion, and migration, and inhibited apoptosis of SGC7901 cells by downregulating the level of SOCS1. In conclusion, miR-922 may have potential for diagnosis of GC.

Phosphorylated Hexa-Acyl Disaccharides Augment Host Resistance Against Common Nosocomial Pathogens.

OBJECTIVES: To determine whether synthetic phosphorylated hexa-acyl disaccharides provide antimicrobial protection in clinically relevant models of bacterial infection. DESIGN: Laboratory study. SETTING: University laboratory. SUBJECTS: BALB/c, C57BL/10J, and C57BL/10ScNJ mice. INTERVENTIONS: Mice were treated with lactated Ringer's (vehicle) solution, monophosphoryl lipid A, or phosphorylated hexa-acyl disaccharides at 48 and 24 hours prior to intraperitoneal Pseudomonas aeruginosa or IV Staphylococcus aureus infection. Leukocyte recruitment, cytokine production, and bacterial clearance were measured 6 hours after P. aeruginosa infection. In the systemic S. aureus infection model, one group of mice was monitored for 14-day survival and another for S. aureus tissue burden at 3 days postinfection. Duration of action for 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide was determined at 3, 10, and 14 days using a model of intraperitoneal P. aeruginosa infection. Effect of 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide on in vivo leukocyte phagocytosis and respiratory burst was examined. Leukocyte recruitment, cytokine production, and bacterial clearance were measured after P. aeruginosa infection in wild-type and toll-like receptor 4 knockout mice treated with 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide or vehicle to assess receptor specificity. MEASUREMENTS AND MAIN RESULTS: During intraperitoneal P. aeruginosa infection, phosphorylated hexa-acyl disaccharides significantly attenuated infection-induced hypothermia, augmented leukocyte recruitment and bacterial clearance, and decreased cytokine production. At 3 days post S. aureus infection, bacterial burden in lungs, spleen, and kidneys was significantly decreased in mice treated with monophosphoryl lipid A or phosphorylated hexa-acyl disaccharides, which was associated with improved survival. Leukocyte phagocytosis and respiratory burst functions were enhanced after treatment with monophosphoryl lipid A or phosphorylated hexa-acyl disaccharides. A time course study showed that monophosphoryl lipid A- and 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide-mediated protection against P. aeruginosa lasts for up to 10 days. Partial loss of augmented innate antimicrobial responses was observed in toll-like receptor 4 knockout mice treated with 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide. CONCLUSIONS: Phosphorylated hexa-acyl disaccharides significantly augment resistance against clinically relevant Gram-negative and Gram-positive infections via enhanced leukocyte recruitment, phagocytosis, and respiratory burst functions of innate leukocytes. Improved antimicrobial protection persists for up to 10 days and is partially mediated through toll-like receptor 4.

Predicting the Health Status of an Unmanned Aerial Vehicles Data-Link System Based on a Bayesian Network.

Unmanned aerial vehicles (UAVs) require data-link system to link ground data terminals to the real-time controls of each UAV. Consequently, the ability to predict the health status of a UAV data-link system is vital for safe and efficient operations. The performance of a UAV data-link system is affected by the health status of both the hardware and UAV data-links. This paper proposes a method for predicting the health state of a UAV data-link system based on a Bayesian network fusion of information about potential hardware device failures and link failures. Our model employs the Bayesian network to describe the information and uncertainty associated with a complex multi-level system. To predict the health status of the UAV data-link, we use the health status information about the root node equipment with various life characteristics along with the health status of the links as affected by the bit error rate. In order to test the validity of the model, we tested its prediction of the health of a multi-level solar-powered unmanned aerial vehicle data-link system and the result shows that the method can quantitatively predict the health status of the solar-powered UAV data-link system. The results can provide guidance for improving the reliability of UAV data-link system and lay a foundation for predicting the health status of a UAV data-link system accurately.

Modeling of BN Lifetime Prediction of a System Based on Integrated Multi-Level Information.

Predicting system lifetime is important to ensure safe and reliable operation of products, which requires integrated modeling based on multi-level, multi-sensor information. However, lifetime characteristics of equipment in a system are different and failure mechanisms are inter-coupled, which leads to complex logical correlations and the lack of a uniform lifetime measure. Based on a Bayesian network (BN), a lifetime prediction method for systems that combine multi-level sensor information is proposed. The method considers the correlation between accidental failures and degradation failure mechanisms, and achieves system modeling and lifetime prediction under complex logic correlations. This method is applied in the lifetime prediction of a multi-level solar-powered unmanned system, and the predicted results can provide guidance for the improvement of system reliability and for the maintenance and protection of the system.

Comparative Transcriptome Profiles of Human Blood in Response to the Toll-like Receptor 4 Ligands Lipopolysaccharide and Monophosphoryl Lipid A.

Monophosphoryl lipid A (MPLA), a less toxic derivative of lipopolysaccharide (LPS), is employed as a vaccine adjuvant and is under investigation as a non-specific immunomodulator. However, the differential response of human leukocytes to MPLA and LPS has not been well characterized. The goal of this study was to compare the differential transcriptomic response of human blood to LPS and MPLA. Venous blood from human volunteers was stimulated with LPS, MPLA or vehicle. Gene expression was determined using microarray analysis. Among 21,103 probes profiled, 136 and 130 genes were differentially regulated by LPS or MPLA, respectively. Seventy four genes were up-regulated and 9 were down-regulated by both ligands. The remaining genes were differentially induced by either agent. Ingenuity Pathway Analysis predicted that LPS and MPLA share similar upstream regulators and have comparable effects on canonical pathways and cellular functions. However, some pro-inflammatory cytokine and inflammasome-associated transcripts were more strongly induced by LPS. In contrast, only the macrophage-regulating chemokine CCL7 was preferentially up-regulated by MPLA. In conclusion, LPS and MPLA induce similar transcriptional profiles. However, LPS more potently induces pro-inflammatory cytokine and inflammasome-linked transcripts. Thus, MPLA is a less potent activator of the pro-inflammatory response but retains effective immunomodulatory activity.

IL-15 Enables Septic Shock by Maintaining NK Cell Integrity and Function.

Interleukin 15 is essential for the development and differentiation of NK and memory CD8+ (mCD8+) T cells. Our laboratory previously showed that NK and CD8+ T lymphocytes facilitate the pathobiology of septic shock. However, factors that regulate NK and CD8+ T lymphocyte functions during sepsis are not well characterized. We hypothesized that IL-15 promotes the pathogenesis of sepsis by maintaining NK and mCD8+ T cell integrity. To test our hypothesis, the pathogenesis of sepsis was assessed in IL-15-deficient (IL-15 knockout, KO) mice. IL-15 KO mice showed improved survival, attenuated hypothermia, and less proinflammatory cytokine production during septic shock caused by cecal ligation and puncture or endotoxin-induced shock. Treatment with IL-15 superagonist (IL-15 SA, IL-15/IL-15Rα complex) regenerated NK and mCD8+ T cells and re-established mortality of IL-15 KO mice during septic shock. Preventing NK cell regeneration attenuated the restoration of mortality caused by IL-15 SA. If given immediately prior to septic challenge, IL-15-neutralizing IgG M96 failed to protect against septic shock. However, M96 caused NK cell depletion if given 4 d prior to septic challenge and conferred protection. IL-15 SA treatment amplified endotoxin shock, which was prevented by NK cell or IFN-γ depletion. IL-15 SA treatment also exacerbated septic shock caused by cecal ligation and puncture when given after the onset of sepsis. In conclusion, endogenous IL-15 does not directly augment the pathogenesis of sepsis but enables the development of septic shock by maintaining NK cell numbers and integrity. Exogenous IL-15 exacerbates the severity of sepsis by activating NK cells and facilitating IFN-γ production.

Flt3 Ligand Treatment Attenuates T Cell Dysfunction and Improves Survival in a Murine Model of Burn Wound Sepsis.

INTRODUCTION: Sepsis is a leading cause of death among severely burned patients. Burn injury disrupts the protective skin barrier and causes immunological dysfunction. In our previous studies, we found that burn injury and wound infection causes a significant decline in lymphocyte populations, implying adaptive immune system dysfunction. In the present study, we examined the effect of treatment with Fms-like tyrosine kinase-3 Ligand (Flt3L) on T cell phenotype and function in a model of burn wound sepsis. FLt3L is an essential cytokine required for hematopoietic progenitor cell development and expansion of both myeloid and lymphoid lineages. Flt3L has been shown to potentiate innate immune functions of dendritic cells and neutrophils during burn wound sepsis. However, the ability of Flt3L to improve T cell function during burn wound sepsis has not been previously evaluated. METHODS: Mice underwent 35% total body surface area scald burn and were treated with Flt3L (10 µg) or vehicle daily via the intraperitoneal route starting 1 day after burn injury. On day 4 after burn injury, Pseudomonas aeruginosa was used to induce wound infection. Leukocytes in spleen and wound draining lymph nodes were characterized using flow cytometry. Bacterial clearance, organ injury, and survival were also assessed. RESULTS: Flt3L treatment prevented the decline in splenic CD4 and CD8 T cells caused by burn injury and infection. Flt3L treatment also attenuated the decline in CD28 expression on CD4 and CD8 T cells and IFNγ production by CD8 T cells in the spleen and wound draining lymph nodes. Furthermore, Flt3L decreased the levels of programmed death ligand 1 expression on splenic dendritic cells and macrophages. Flt3 treatment improved systemic bacterial clearance, decreased liver and kidney injury, and significantly improved survival in mice with burn wound sepsis. CONCLUSION: Burn injury and associated sepsis causes significant loss of T cells and evidence of T cell dysfunction. Flt3L attenuates T cell dysfunction and improves host resistance to burn wound sepsis in mice.

The role of MyD88- and TRIF-dependent signaling in monophosphoryl lipid A-induced expansion and recruitment of innate immunocytes.

Treatment with the TLR4 agonist MPLA augments innate resistance to common bacterial pathogens. However, the cellular and molecular mechanisms by which MPLA augments innate immunocyte functions are not well characterized. This study examined the importance of MyD88- and TRIF-dependent signaling for leukocyte mobilization, recruitment, and activation following administration of MPLA. MPLA potently induced MyD88- and TRIF-dependent signaling. A single injection of MPLA caused rapid mobilization and recruitment of neutrophils, a response that was largely mediated by the chemokines CXCL1 and -2 and the hemopoietic factor G-CSF. Rapid neutrophil recruitment and chemokine production were regulated by both pathways although the MyD88-dependent pathway showed some predominance. In further studies, multiple injections of MPLA potently induced mobilization and recruitment of neutrophils and monocytes. Neutrophil recruitment after multiple injections of MPLA was reliant on MyD88-dependent signaling, but effective monocyte recruitment required activation of both pathways. MPLA treatment induced expansion of myeloid progenitors in bone marrow and upregulation of CD11b and shedding of L-selectin by neutrophils, all of which were attenuated in MyD88- and TRIF-deficient mice. These results show that MPLA-induced neutrophil and monocyte recruitment, expansion of bone marrow progenitors and augmentation of neutrophil adhesion molecule expression are regulated by both the MyD88- and TRIF-dependent pathways.

Identification of less lipophilic riminophenazine derivatives for the treatment of drug-resistant tuberculosis.

Clofazimine (CFZ), a member of the riminophenazine class, has been studied in clinical trials for the treatment of multidrug-resistant tuberculosis (MDR-TB). CFZ has several side effects which can be attributed to its extremely high lipophilicity. A series of novel riminophenazine analogues bearing a C-2 pyridyl substituent was designed and synthesized with the goal of maintaining potent activity against Mycobacterium tuberculosis (M. tuberculosis) while improving upon its safety profile by lowering the lipophilicity. All compounds were evaluated for their in vitro activity and cytotoxicity. The results demonstrated that many new compounds had potent activity against M. tuberculosis with MICs of less than 0.03 µg/mL and low cytotoxicity with IC(50) values greater than 64 µg/mL. Some compounds were tested for in vivo efficacy against MDR-TB in an experimental mouse infection model. Two compounds demonstrated equivalent or better efficacy than CFZ in this model with significantly reduced skin discoloration potential.

[Effects of lactational dioxin exposure to development of alveolar bone in SD rat offspring].

OBJECTIVE: To study the influence of lactational dioxin exposure (2, 3, 7, 8-tetrachlorodibenzo-p-dixon, TCDD) on development of alveolar bone in SD rat offspring. METHODS: The rats of TCDD exposure group and control group were sacrificed and the alveolar bone with molars of PD60 rats in the two groups were embedded in resin. The sections were observed by fluorescent microscope. The alveolar bone formation was evaluated by histological examination, tetracycline fluorescence marker and quantitative histomorphometry. The indices of quantitative histomorphometry were compared. RESULTS: The trabecular structure of alveolar bone was looser in TCDD exposure group than in the control group. The tetracycline fluorescence markers were more disorganized in TCDD group. The indices of quantitative histomorphometry of alveolar bone between two groups showed significantly different. CONCLUSIONS: Lactational 2,3,7,8-TCDD exposure decreased the quality and quantity of alveolar bone in SD rat offspring. It is suggested that dioxins exposure may interrupt the spatial configuration.

Detecting objects of variable shape structure with hidden state shape models.

This paper proposes a method for detecting object classes that exhibit variable shape structure in heavily cluttered images. The term "variable shape structure" is used to characterize object classes in which some shape parts can be repeated an arbitrary number of times, some parts can be optional, and some parts can have several alternative appearances. Hidden State Shape Models (HSSMs), a generalization of Hidden Markov Models (HMMs), are introduced to model object classes of variable shape structure using a probabilistic framework. A polynomial inference algorithm automatically determines object location, orientation, scale and structure by finding the globally optimal registration of model states with the image features, even in the presence of clutter. Experiments with real images demonstrate that the proposed method can localize objects of variable shape structure with high accuracy. For the task of hand shape localization and structure identification, the proposed method is significantly more accurate than previously proposed methods based on chamfer-distance matching. Furthermore, by integrating simple temporal constraints, the proposed method gains speed-ups of more than an order of magnitude, and produces highly accurate results in experiments on non-rigid hand motion tracking.

Pulmonary fissure segmentation on CT.

A pulmonary fissure is a boundary between the lobes in the lungs. Its segmentation is of clinical interest as it facilitates the assessment of lung disease on a lobar level. This paper describes a new approach for segmenting the major fissures in both lungs on thin-section computed tomography (CT). An image transformation called "ridge map" is proposed for enhancing the appearance of fissures on CT. A curve-growing process, modeled by a Bayesian network, is described that is influenced by both the features of the ridge map and prior knowledge of the shape of the fissure. The process is implemented in an adaptive regularization framework that balances these influences and reflects the causal dependencies in the Bayesian network using an entropy measure. The method effectively alleviates the problem of inappropriate weights of regularization terms, an effect that can occur with static regularization methods. The method was applied to segment and visualize the lobes of the lungs on chest CT of 10 patients with pulmonary nodules. Only 78 out of 3286 left or right lung regions with fissures (2.4%) required manual correction. The average distance between the automatically segmented and the manually delineated "ground-truth" fissures was 1.01 mm, which was similar to the average distance of 1.03 mm between two sets of manually segmented fissures. The method has a linear-time worst-case complexity and segments the upper lung from the lower lung on a standard computer in less than 5 min.

[The effect of lead exposure in utero on the teeth eruption and enamel development of rat offspring.].

OBJECTIVE: To investigate the effects of lead exposure at different levels in utero on the teeth eruption and enamel development of rat offsprings. METHODS: 27 pregnant SD rats were divided into three groups randomly: high level lead group (HLG), low level lead group (LLG) and control group with nine rats in each group. The three groups from the gestation day to the end of the gestation were given either deionized water in control group or deionized water containing 200 mg/L Pb2+ as lead acetate through drinking method in high level lead experimental group and 50 mg/L Pb2+ as lead acetate through drinking method in low level lead experimental group. The incisors of newborn rats were marked at the level of the gingival papilla on the 26th day after birth. On the 36th day, the incisors of newborn rats were marked again at the same level. Then the rat offsprings were killed and their blood was collected for lead analysis. The mandible incisors of rat offspring were separated and the content of Pb in incisors was determined by using a graphite furnace atomic absorption spectrometric method. The teeth of rat offspring were observed and the distance between two marks were measured by means of stereomicroscope. The ratio of calcium to phosphate of enamel of rat offspring was compared by electron probe microanalyses. RESULTS: The level of blood lead in 200 mg/L, 50 mg/L treated rat offspring groups was higher than that in control group. The tooth lead of 200 mg/L, 50 mg/L treated rat offspring groups [(77.3 +/- 6.3), (27.8 +/- 4.5) microg/g] were higher than the control [(6.6 +/- 0.8) microg/g, P < 0.01]. Compared with the control group, the teeth of lead exposure experimental groups were smaller and severity of attrition was obvious and pulpal perforations were often observed. These appearances was more distinct in rats of high level lead experimental group. The incisors of lead-treated rat offspring erupted [(0.25 +/- 0.08), (0.30 +/- 0.09) mm/d] more slowly than control ones [(0.39 +/- 0.09) mm/d, P < 0.01]. The ratio of calcium to phosphate (Ca/P) decreased with the increase of lead exposure. It was found that Ca/P in lead exposure experimental groups (1.68 +/- 0.54), (1.37 +/- 0.47) was significantly lower than that in the control group (2.14 +/- 0.33). CONCLUSION: Lead exposure in utero affects the normal eruption of teeth and enamel formation and the degree was related with the lead exposure level.

Segmentation of nodules on chest computed tomography for growth assessment.

Several segmentation methods to evaluate growth of small isolated pulmonary nodules on chest computed tomography (CT) are presented. The segmentation methods are based on adaptively thresholding attenuation levels and use measures of nodule shape. The segmentation methods were first tested on a realistic chest phantom to evaluate their performance with respect to specific nodule characteristics. The segmentation methods were also tested on sequential CT scans of patients. The methods' estimation of nodule growth were compared to the volume change calculated by a chest radiologist. The best method segmented nodules on average 43% smaller or larger than the actual nodule when errors were computed across all nodule variations on the phantom. Some methods achieved smaller errors when examined with respect to certain nodule properties. In particular, on the phantom individual methods segmented solid nodules to within 23% of their actual size and nodules with 60.7 mm3 volumes to within 14%. On the clinical data, none of the methods examined showed a statistically significant difference in growth estimation from the radiologist.