AIM2 enhances Candida albicans infection through promoting macrophage apoptosis via AKT signaling.

Candida albicans is among the most prevalent invasive fungal pathogens for immunocompromised individuals and novel therapeutic approaches that involve immune response modulation are imperative. Absent in melanoma 2 (AIM2), a pattern recognition receptor for DNA sensing, is well recognized for its involvement in inflammasome formation and its crucial role in safeguarding the host against various pathogenic infections. However, the role of AIM2 in host defense against C. albicans infection remains uncertain. This study reveals that the gene expression of AIM2 is induced in human and mouse innate immune cells or tissues after C. albicans infection. Furthermore, compared to their wild-type (WT) counterparts, Aim2-/- mice surprisingly exhibit resistance to C. albicans infection, along with reduced inflammation in the kidneys post-infection. The resistance of Aim2-/- mice to C. albicans infection is not reliant on inflammasome or type I interferon production. Instead, Aim2-/- mice display lower levels of apoptosis in kidney tissues following infection than WT mice. The deficiency of AIM2 in macrophages, but not in dendritic cells, results in a phenocopy of the resistance observed in Aim2-/- mice against C. albican infection. The treatment of Clodronate Liposome, a reagent that depletes macrophages, also shows the critical role of macrophages in host defense against C. albican infection in Aim2-/- mice. Furthermore, the reduction in apoptosis is observed in Aim2-/- mouse macrophages following infection or treatment of DNA from C. albicans in comparison with controls. Additionally, higher levels of AKT activation are observed in Aim2-/- mice, and treatment with an AKT inhibitor reverses the host resistance to C. albicans infection. The findings collectively demonstrate that AIM2 exerts a negative regulatory effect on AKT activation and enhances macrophage apoptosis, ultimately compromising host defense against C. albicans infection. This suggests that AIM2 and AKT may represent promising therapeutic targets for the management of fungal infections.

Tumour-associated antigens in systemic lupus erythematosus: association with clinical manifestations and serological indicators.

OBJECTIVES: To explore the relationship of tumour-associated antigens (TAAs) with the clinical manifestations and serological markers of SLE. METHODS: This was a retrospective study. Clinical data of SLE patients were extracted from the electronic medical records, including serum levels of TAAs such as alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), cancer antigen (CA) 19-9, CA125, CA15-3 and cytokeratin 19-fragments (CYFRA21-1). TAA positivity was defined as serum level exceeding the upper limit of the corresponding reference range. RESULTS: A total of 149 SLE patients (SLE group) and 149 age- and sex-matched healthy subjects (control group) were enrolled. Compared with healthy controls, the SLE group had higher positivity rates for CA19-9 and CYFRA21-1, and elevated serum levels of CA125, CA15-3 and CYFRA21-1. SLE patients with TAA positivity were older, had a higher prevalence of serous effusion, pericardial effusion, albuminuria and thrombocytopenia, and lower positivity rate for anti-dsDNA than patients without TAA positivity. The levels of serum creatinine (SCR), blood urea nitrogen, glutamic oxalate transaminase and 24-h urinary protein were also higher in SLE patients with TAA positivity, but platelet count and serum albumin levels were lower. On logistic regression, thrombocytopenia and SCR levels were identified as independent risk factors for TAA positivity. CA125 positivity rate and serum levels of CA125 were associated with SLE disease activity. CONCLUSION: The positivity rates and serum levels of some TAAs were elevated in SLE, and thrombocytopenia and SCR levels were independent risk factors for TAA positivity.

Time-dependent changes in RPILD and mortality risk in anti-MDA5+ DM patients: a cohort study of 272 cases in China.

OBJECTIVES: Anti-melanoma differentiation-associated gene 5 positive (anti-MDA5+) DM has a close relationship with rapidly progressive interstitial lung disease (RPILD) and is associated with high mortality. However, data regarding the time-dependent risk of RPILD and deaths during disease progression are limited. We conducted this study to investigate whether the risk of RPILD and death were time-dependent or not in anti-MDA5+ DM. METHODS: We assessed a cohort of 272 patients with anti-MDA5+ DM. The clinical characteristics of patients with anti-MDA5+ were collected, and COX regression was used to analyse independent risk factors for RPILD and death. We also described changes in risk of RPILD and death over time and their potential clinical implications. RESULTS: There were 272 anti-MDA5+ DM patients enrolled in this study. According to the multivariate cox regression analysis, short disease course, high CRP level, anti-Ro52 positive and anti-MDA5 titre (++∼+++) were independent risk factors of RPILD. High creatine kinase level, high CRP level and RPILD were independent risk factors for death, and >90% RPILD and 84% mortality occurred in the first 6 months after disease onset. Notably, the first 3 months is a particularly high-risk period, with 50% of RPILD and 46% of deaths occurring. Hazards regarding RPILD and mortality diminished over time during a median follow-up of 12 months. CONCLUSION: These results suggest significant, time-dependent changes in RPILD and mortality risk in anti-MDA5+ DM patients, providing a cut-off time window to estimate disease progression and poor prognosis.

Nanoscale Thin-Film Boiling Processes on Heterogeneous Surfaces.

Acquiring rapid and efficient boiling processes has been the focus of industry as they have the potential to improve the energy efficiency and reduce the carbon emissions of production processes. Here, we report nanoscale thin-film boiling on different heterogeneous surfaces. Through nonequilibrium molecular dynamics simulation, we captured the triple-phase interface details, visualized the bubble nucleation, and recorded the internal fluid flow and thermal characteristics. It is found that nanoscale thin-film boiling without the occurrence of bubble nucleation shows excellent heat and mass transfer performance, which differs from macroscale boiling. In general, rough structures advance the onset time of stable boiling and improve the efficiency. The heat transfer coefficient and heat flux on a rough hydrophilic surface respectively reach to 7.43 × 104 kW/(m2·K) and 1.3 × 106 kW/m2 at a surface temperature of 500 K, which are 100-fold higher than those of micrometer-scale thin-film boiling. However, due to the resultant vapor film trapped between the liquid and the surface, the rough hydrophobic surface leads to heat transfer deterioration instead. It is revealed that the underlying mechanism of regulatory effects resulting from surface physicochemical properties is originated from the variation of interfacial thermal resistance. It is available to reduce the overall interfacial resistance and further improve the heat and mass transfer efficiency through increasing surface roughness, enhancing surface wettability, and increasing the area proportion of the hydrophilic region. This work provides guidelines to achieve rapid and efficient thin-liquid-film boiling and serves as a reference for the optimized design of surfaces utilized for high-heat flux removal through vaporization processes.

Iguratimod alleviates tubulo-interstitial injury in mice with lupus.

INTRODUCTION: Tubulo-interstitial injury is a poor prognostic factor for lupus nephritis (LN). Here, we tested whether iguratimod could inhibit tubulo-interstitial injury in LN. METHODS: MRL/lpr mice, an animal model of lupus, were treated with iguratimod or vehicle solution. Pathological changes of kidney were evaluated blindly by the same pathologist. Renal type I collagen (COL-I), IgG, E-cadherin, fibroblast-specific protein 1 (FSP-1) were detected by immunofluorescence, immunohistochemical staining or quantitative real-time PCR. After treated with transforming growth factor ß1 (TGF-ß1) and iguratimod, E-cadherin, fibronectin, Smad2/3, p38 MAPK, p-Smad2/3, and p-p38 MAPK, ß-catenin and TGF-ß type II receptor (TGFßRII) in HK2 cells were measured by western blotting, quantitative real-time PCR or immunofluorescence. RESULTS: Iguratimod reduced immune deposition along the tubular basement membrane, inhibited the tubulo-interstitial infiltration of inflammatory cells, and alleviated tubular injury in MRL/lpr mice. Moreover, Iguratimod eased the tubulo-interstitial deposition of collagen fibers, which was confirmed by decreased expression of COL-I. Furthermore, iguratimod suppressed the expression of FSP-1 and increased that of E-cadherin in renal tubular epithelial cells. In HK2 cells cultured with TGF-ß1, iguratimod treatment not only reversed cellular morphological changes, but also prevented E-cadherin downregulation and fibronectin upregulation. In addition, iguratimod inhibited phosphorylation of TGFßRII, Smad2/3 and p38 MAPK in HK2 cells treated with TGF-ß1, and also blocked nuclear translocation of ß-catenin. CONCLUSION: Iguratimod eased tubulo-interstitial lesions in LN, especially tubulo-interstitial fibrosis, and might have potential as a drug for inhibiting the progression of tubulo-interstitial fibrosis in LN.

Head-on Collision of Two Nanodroplets on a Solid Surface: A Molecular Dynamics Simulation Study.

Most researchers focus on the collision of a single droplet with a solid surface, while it is common for a droplet to collide with a sessile droplet on a solid surface in reality. This study performed the head-on collision of two nanodroplets on a solid surface using the molecular dynamics simulation method. The effects of impact velocity, interaction intensity between solid and liquid atoms, and the solid fraction of the surface on the collision process are studied with independent simulation cases. The maximum spreading factor and the dimensionless maximum spreading time are recorded and calculated to describe the collision process quantitatively. The simulation results indicate that the maximum spreading factor depends more on the solid fraction than the interaction intensity since it does not fundamentally change the wetting state of the droplet at its maximum spreading state. Because of two different effects, the maximum dimensionless spreading time decreases first and then increases with the interaction intensity, and both effects weaken with the increase of impact velocity. As the solid fraction increases, the maximum spreading factor increases significantly at high impact velocity, and the maximum dimensionless spreading time first decreases and then increases because the wetting state of the coalescent droplet at the maximum spreading moment gradually changes from the Wenzel state to the Cassie state. In general, the initial wetting state of the sessile droplet and the wetting state of the coalescent droplet at the maximum spreading moment have important effects on the maximum spreading factor and the maximum spreading time. We establish a theoretical prediction model for the maximum spreading factor on a smooth surface based on energy conservation with quite good accuracy. This research has improved our understanding of the head-on collision process of two nanodroplets on a solid surface.

Evaporation-Induced Wetting Transition of Nanodroplets on Nanopatterned Surfaces with Concentric Rings: Surface Geometry and Wettability Effects.

Droplet evaporation is widespread in natural and industrial application, and the rapid and efficient evaporation can significantly improve energy efficiency. However, the fundamental mechanism of contact line dynamics and the microscopic characteristics of evaporating nanodroplets are not well understood. Moreover, how to design a nanostructure surface to enhance nanodroplet evaporation remains unclear. Here, through molecular dynamics simulation, we investigated the evaporation dynamics of nanodroplets on various nanoring surfaces with different geometric parameters and wettability. By measuring the changes of contact radius and contact angle, the results showed that nanodroplets successively exhibit constant contact angle (CCA), constant contact radius (CCR), and mix mode during evaporation, and the evaporation-induced CCA-CCR transition, in essence, is a Cassie-Wenzel wetting transition, whose onset time is remarkably dependent on the surface roughness and wettability. We found that this evaporation-induced wetting transition is postponed on the surface with small nanostructure spacing and weak hydrophilicity, and the evaporation rate of nanodroplets improves accordingly. The dense and hydrophobic nanostructures can not only restrain the Cassie-Wenzel transition, but also enhance the evaporation rate of nanodroplets. Last, through the potential energy field analysis of nanoring substrates, we revealed that the Cassie-Wenzel wetting transition of nanodroplets is a process of molecule migration to low potential energy regions. Our work provides guidance for designing nanostructure surfaces to effectively control the droplet wetting state and enhance its mass transfer performance of phase change.

Anticoagulant and Antithrombotic Properties in Vitro and in Vivo of a Novel Sulfated Polysaccharide from Marine Green Alga Monostroma nitidum.

Sulfated polysaccharides from marine algae have high potential as promising candidates for marine drug development. In this study, a homogeneous sulfated polysaccharide from the marine green alga Monostroma nitidum, designated MS-1, was isolated using water extraction and anion-exchange and size-exclusion chromatography. Results of chemical and spectroscopic analyses showed that MS-1 mainly consisted of →3)-α-l-Rhap-(1→ and →2)-α-l-Rhap-(1→ residues, with additional branches consisting of 4-linked ß-d-xylose, 4-/6-linked d-glucose, terminal ß-d-glucuronic acid, and 3-/2-linked α-l-rhamnose. Sulfate ester groups substituted mainly at C-2/C-4 of →3)-α-l-Rhap-(1→ and C-4 of →2)-α-l-Rhap-(1→ residues, slightly at C-2 of terminal ß-d-glucuronic residues. MS-1 exhibited strong anticoagulant activity in vitro and in vivo as evaluated by the activated partial thromboplastin time and thrombin time assays, and significantly decreased platelet aggregation. The anticoagulant activity mechanism of MS-1 was mainly attributed to strong potentiation thrombin by heparin cofactor-II, and it also hastened thrombin and coagulation factor Xa inhibitions by potentiating antithrombin-III. MS-1 possessed markedly thrombolytic activity evaluated by plasminogen activator inhibitior-1, fibrin degradation products, and D-dimer levels using rats plasma, and recanalization rate by FeCl3-induced carotid artery thrombosis in mice. MS-1 exhibited strong antithrombotic activity in vitro and in vivo evaluated by the wet weighs and lengths of thrombus, and thrombus occlusion time by electrically-induced carotid artery thrombosis in rats. These results suggested that MS-1 could be a promising marine drug for prevention and therapy of thromboembolic disease.

Inhibition of maternally expressed gene 3 attenuated lipopolysaccharide-induced apoptosis through sponging miR-21 in renal tubular epithelial cells.

Acute kidney injury (AKI) results in retention of waste products and dysregulation of extracellular volume and electrolytes, thus leading to a variety of complications. Recent advances in long noncoding RNAs suggested their close relationship with disease progression. In the current study, we investigated the role and mechanism of maternally expressed gene 3 (MEG3) on AKI pathogenesis. Real-time polymerase chain reaction found that the expression of MEG3 was significantly increased in both kidney tissues and TKPTS cells induced by lipopolysaccharide (LPS). Western blot assay showed that the expression of apoptosis regulator Bcl-2 was increased in MEG3-inhibited TKPTS cells. Flow cytometry assay confirmed that LPS-induced apoptosis was significantly attenuated after transfection of si-MEG3. The RNAhybrid informatics algorithm predicted that there was a strong binding capacity between miR-21 and MEG3. Luciferase reporter assay confirmed that MEG3 could function as a competing endogenous RNA of miR-21. The antiapoptotic effect of si-MEG3 could be neutralized by a miR-21 inhibitor, demonstrated by the decreased expression of Bcl-2 and flow cytometry results. Further investigation showed that programmed cell death protein 4 (PDCD4), a validated target of miR-21, was highly expressed in both injured kidney tissues and LPS-stimulated TKPTS cells. Meanwhile, the protein expression of PDCD4 was significantly reduced by inhibition of MEG3, but retrieved by coinhibition of MEG3 and miR-21. In conclusion, our results demonstrated that inhibition of MEG3 could attenuate LPS-induced apoptosis in TKPTS cells by regulating the miR-21/PDCD4 pathway, suggesting that the MEG3/miR-21/PDCD4 axis could be developed as a potential therapeutic target of AKI.

Nanodroplets Impact on Rough Surfaces: A Simulation and Theoretical Study.

Impact of droplets is widespread in life, and modulating the dynamics of impinging droplets is a significant problem in production. However, on textured surfaces, the micromorphologic change and mechanism of impinging nanodroplets are not well-understood; furthermore, the accuracy of the theoretical model for nanodroplets needs to be improved. Here, considering the great challenge of conducting experiments on nanodroplets, a molecular dynamics simulation is performed to visualize the impact process of nanodroplets on nanopillar surfaces. Compared with macroscale droplets, apart from the similar relation of restitution coefficient with the Weber number, we found some distinctive results: the maximum spreading time is described as a power law of impact velocity, and the relation of maximum spreading factor with impact velocity or the Reynolds number is exponential. Moreover, the roughness of substrates plays a prominent role in the dynamics of impact nanodroplets, and on surfaces with lower solid fraction, the lower attraction force induces an easier rebound of impact nanodroplets. At last, on the basis of the energy balance, through modifying the estimation of viscous dissipation and surface energy terms, we proposed an improved model for the maximum spreading factor, which shows greater accuracy for nanodroplets, especially in the low-to-moderate velocity range. The outcome of this study demonstrates that a distinctive dynamical behavior of impinging nanodroplets, the fundamental insight, and more accurate prediction are very useful in the improvement of the hydrodynamic behavior of the nanodroplets.

Reverse electrodialysis in bilayer nanochannels: salinity gradient-driven power generation.

To evaluate the possibility of nano-fluidic reverse electrodialysis (RED) for salinity gradient energy harvesting, we consider the behavior of ion transportation in a bilayer cylindrical nanochannel consisting of different sized nanopores connecting two large reservoirs at different NaCl concentrations. Numerical simulations to illustrate the electrokinetic behavior at asymmetric sub-pore length and surface charge density are conducted, the impacts of which on transference number, osmotic current, diffusive voltage, maximum power and maximum power efficiency are systematically investigated. The results reveal that the transference number in Config. I (where high NaCl concentration is applied at the larger nanopore) is always larger than that in the opposite configuration (Config. II). At low concentration ratios, the osmotic current and maximum power have maximum values, while the maximum power efficiency decreases consistently. For Config. II, the ion transportation is impacted by the surface charge density at both sub-nanopores, while for Config. I, it is determined by the surface charge density at the downstream small nanopore. When large surface charge density is applied at the downstream small nanopore in contact with a very low concentration reservoir, there exists an interesting phenomenon: the larger surface charge density at the large nanopore induces a slight performance drop due to the impact of upstream EDL overlap.

Effects of Solid Fraction on Droplet Wetting and Vapor Condensation: A Molecular Dynamic Simulation Study.

Recently, numerous studies focused on the wetting process of droplets on various surfaces at a microscale level. However, there are a limited number of studies about the mechanism of condensation on patterned surfaces. The present study performed the dynamic wetting behavior of water droplets and condensation process of water molecules on substrates with different pillar structure parameters, through molecular dynamic simulation. The dynamic wetting results indicated that droplets exhibit Cassie state, PW state, and Wenzel state successively on textured surfaces with decreasing solid fraction. The droplets possess a higher static contact angle and a smaller spreading exponent on textured surfaces than those on smooth surfaces. The condensation processes, including the formation, growth, and coalescence of a nanodroplet, are simulated and quantitatively recorded, which are difficult to be observed by experiments. In addition, a wetting transition and a dewetting transition were observed and analyzed in condensation on textured surfaces. Combining these simulation results with previous theoretical and experimental studies will guide us to understand the hypostasis and mechanism of the condensation more clearly.

Iron deficiency anemia's effect on bone formation in zebrafish mutant.

Iron is one of the essential elements of life. Iron metabolism is related to bone metabolism. Previous studies have confirmed that iron overload is a risk factor for osteoporosis. But the correlation between iron deficiency and bone metabolism remains unclear. Ferroportin 1 is identified as a cellular iron exporter and required for normal iron cycling. In zebrafish, the mutant of ferroportin 1 gene (fpn1), weh(tp85c) exhibited the defective iron transport, leading to developing severe hypochromic anemia. We used weh(tp85c) as a model for investigating iron deficiency and bone metabolism. In this study, we examined the morphology of the developing cartilage and vertebrae of the Weh(tp85) compared to the wild type siblings by staining the larvae with alcian blue for cartilage and alizarin red for the bone. In addition, we evaluated the expression patterns of the marker genes of bone development and cell signaling in bone formation. Our results showed that weh(tp85c) mutant larvae exhibited the defects in bone formation, revealing by decreases in the number of calcified vertebrae along with decreased expression of osteoblast novel genes: alpl, runx2a and col1a1a and BMPs signaling genes in osteoblast differentiation: bmp2a and bmp2b. Our data suggest that iron deficiency anemia affects bone formation, potentially through the BMPs signaling pathway in zebrafish.

Estrogen-induced expression of tumor necrosis factor-like weak inducer of apoptosis through ERα accelerates the progression of lupus nephritis.

OBJECTIVES: Oestrogens have been shown to play key roles in the pathogenesis of SLE. The aim of this study was to investigate the roles and mechanisms of 17ß-estradiol (E2) in TNF-like weak inducer of apoptosis (TWEAK) expression in LN. METHODS: Peripheral blood mononuclear cells (PBMCs) obtained from LN patients were used for in vitro experiments, while female MRL/lpr and MRL/MpJ mice were used for in vivo studies. E2, ICI 182 780 [estrogen receptor (ER)-selective antagonist], methyl-piperidino-pyrazole (MPP, ERα-selective modulator), lentivirus (LV)-TWEAK-short hairpin RNA (shRNA) and LV-control-shRNA treatments were used in this study. RESULTS: TWEAK mRNA expression in PBMCs was significantly increased following E2 treatment and downregulated after incubation with ICI 182 780 or MPP. Compared with sham-operated MRL/lpr mice, ovariectomized mice, treated with dimethyl sulphoxide vehicle alone, showed lower expression levels of renal TWEAK mRNA and protein. The expression of both mRNA and protein in ovariectomized mice was upregulated after E2 treatment and downregulated after ICI 182 780 or MPP co-treatment. Severe renal damage was observed in E2-treated ovariectomized mice, as were higher serum levels of IL-6, compared with dimethyl sulphoxide vehicle-treated ovariectomized mice. Co-treatment with LV-TWEAK-shRNA reversed these changes, and LV-control-shRNA treatment had no effect on them. CONCLUSION: Our results demonstrated that E2 plays an important role in the upregulation of TWEAK expression in LN, most likely through an ERα-dependent pathway, causing kidney damage. This provides a novel insight into the mechanisms of the E2-TWEAK signalling pathway in LN.

Tumor Necrosis Factor-Like Weak Inducer of Apoptosis Accelerates the Progression of Renal Fibrosis in Lupus Nephritis by Activating SMAD and p38 MAPK in TGF-ß1 Signaling Pathway.

This study aim was to explore the effects of tumor necrosis factor-like weak inducer of apoptosis (TWEAK) in lupus nephritis and its potential underlying mechanisms. MRL/lpr mice were used for in vivo experiments and human proximal tubular cells (HK2 cells) were used for in vitro experiments. Results showed that MRL/lpr mice treated with vehicle solution or LV-Control shRNA displayed significant proteinuria and severe renal histopathological changes. LV-TWEAK-shRNA treatment reversed these changes and decreased renal expressions of TWEAK, TGF-ß1, p-p38 MAPK, p-Smad2, COL-1, and α-SMA proteins. In vitro, hTWEAK treatment upregulated the expressions of TGF-ß1, p-p38 MAPK, p-SMAD2, α-SMA, and COL-1 proteins in HK2 cells and downregulated the expressions of E-cadherin protein, which were reversed by cotreatment with anti-TWEAK mAb or SB431542 treatment. These findings suggest that TWEAK may contribute to chronic renal changes and renal fibrosis by activating TGF-ß1 signaling pathway, and phosphorylation of Smad2 and p38 MAPK proteins was also involved in this signaling pathway.

Value of SLE-DAS in assessing disease activity in patients with systemic lupus erythematosus: a single-centre retrospective study.

OBJECTIVES: This study aimed to evaluate the clinical value of the Systemic Lupus Erythematosus Disease Activity Score (SLE-DAS) for assessing disease activity in patients with SLE. METHODS: Clinical data were collected from patients with SLE who were admitted at the Second Affiliated Hospital of Soochow University from January 2009 to December 2022. The glucocorticoid dose grading was used as the gold standard for disease activity assessment in SLE. The SLE-DAS value was calculated, and the SLE disease activity status was graded based on the SLE-DAS value. Another scoring criterion, the SLE Disease Activity Index 2000 (SLEDAI 2000), served as a control. Spearman correlation analysis was used to calculate the correlation between the scoring criteria and other variables. RESULTS: The analysis included 396 patients with SLE. A strong correlation was found between SLE-DAS and SLEDAI 2000 (ρ=0.709, 95% CI 0.648 to 0.766, p<0.001), with median SLE-DAS and SLEDAI 2000 scores of 15.32 (7.90 to 24.45) and 13 (8 to 19), respectively. Compared with the SLEDAI 2000 value, the SLE-DAS value correlated better with glucocorticoid dose grading (ρ=0.434 vs 0.518), gammaglobulin use (ρ=0.170 vs 0.318) and immunosuppressant use (ρ=0.122 vs 0.221). A moderate correlation based on disease activity grading was found between SLE-DAS and glucocorticoid dose grading (ρ=0.441), whereas a mild correlation was observed between SLEDAI 2000 and glucocorticoid dose grading (ρ=0.325). Additionally, SLE-DAS revealed a positive correlation with severe thrombocytopenia, cardiac involvement and pulmonary involvement but not SLEDAI 2000. CONCLUSION: Compared with SLEDAI 2000, SLE-DAS may provide a more accurate disease activity assessment in patients with SLE, especially those with severe thrombocytopenia and cardiopulmonary involvement.