Outcomes of intensive and nonintensive blast-reduction strategies in accelerated and blast-phase MPN.

ABSTRACT: Transformation of BCR::ABL1-negative myeloproliferative neoplasms (MPN) to an accelerated or blast phase is associated with poor outcomes. The efficacy of acute myeloid leukemia (AML)-type intensive and nonintensive hypomethylating agent-based regimens is not well studied. We therefore performed a retrospective analysis of patients with MPN-AP/BP (N = 138) treated with intensive (N = 81) and nonintensive (N = 57) blast-reduction strategies. We used clinically relatable response criteria developed at the Princess Margaret Cancer Centre. The overall best response, comprising complete remission (CR), complete remission with incomplete hematologic recovery (CRi), and reversion to chronic phase MPN (cMPN), in the intensive and nonintensive groups was 77% (62 of 81) and 39% (21 of 54), respectively. Similar overall best response rates were observed in patients receiving induction with daunorubicin combined with cytarabine arabinoside (daunorubicin + ara-C) (74% [23 of 31]) or FLAG-IDA/NOVE-HiDAC (78% [39 of 50], P = .78). However, patients receiving daunorubicin + ara-C more often required second inductions (29% [9 of 31] vs 4% [2 of 50], P = .002). Most responses in the entire cohort were reversions to cMPN (55 of 83 [66%]). CR and CRi comprised 30% (25 of 83) and 4% (3 of 83) of responses, respectively. Mutations in TP53 (overall response [OR] 8.2 [95% confidence interval [CI] 2.01, 37.1], P = .004) and RAS pathway (OR 5.1 [95%CI 1.2, 23.7], P = .03) were associated with inferior treatment response for intensively treated patients, and poorer performance status (Eastern Cooperative Oncology Group) was associated with inferior treatment response in both intensively (OR 10.4 [95% CI 2.0, 78.5], P = .009) and nonintensively treated groups (OR 12 [95% CI 2.04, 230.3], P = .02). In patients with paired samples before and after therapy (N = 26), there was a significant residual mutation burden remaining irrespective of response to blast-reduction therapy.

Length changes in the distal oblique band during forearm rotation measured using four-dimensional CT: an in vivo study.

Length change in the distal oblique band during forearm rotation was measured using four-dimensional CT in seven volunteers. There was no significant change in length, which provides more theoretical support for distal oblique band reinforcement for treatment of instability of the distal radioulnar joint.

Upfront allogeneic transplantation versus JAK inhibitor therapy for patients with myelofibrosis: a North American collaborative study.

Allogeneic hematopoietic cell transplantation (HCT) is the only curative therapy for myelofibrosis (MF) and is recommended for patients with higher risk disease. However, there is a risk of early mortality, and optimal timing is unknown. JAK inhibitor (JAKi) therapy may offer durable improvement in symptoms, splenomegaly and quality of life. The aim of this multicentre, retrospective observational study was to compare outcomes of patients aged 70 years or below with MF in chronic phase who received upfront JAKi therapy vs. upfront HCT in dynamic international prognostic scoring system (DIPSS)-stratified categories. For the whole study cohort, median overall survival (OS) was longer for patients who received a JAKi vs. upfront HCT, 69 (95% CI 57-89) vs. 42 (95% CI 20-not reached, NR) months, respectively (p = 0.01). In patients with intermediate-2 and high-risk disease, median OS was 55 (95% CI 36-73) months with JAKi vs. 36 (95% CI 20-NR) months for HCT (p = 0.27). An upfront HCT strategy was associated with early mortality and difference in median OS was not observed in any risk group by 5 years of follow-up. Within the limitations of a retrospective observational study, we did not observe any benefit of a universal upfront HCT approach for higher-risk MF.

Differences in the Proportion of CYP2C19 Loss-of-Function Between Cerebral Infarction and Coronary Artery Disease Patients.

Background: Cytochrome P450 2C19 (CYP2C19) genotypes and metabolic phenotypes (extensive metabolizer (EM), intermediate metabolizer (IM), and poor metabolizer (PM)) are related to the metabolism of therapeutic drugs for cardiovascular and cerebrovascular diseases. This study aimed to investigate the differences of CYP2C19 gene polymorphism distribution between coronary artery disease (CAD) patients and cerebral infarction (CI) patients. Methods: We identified 413 CI patients, 509 CAD patients, and 241 CI+CAD patients from 2016 to 2020 and studied genotypes of CYP2C19 rs4986893 (636G>A) and rs4244285 (681G>A) polymorphisms using PCR-gene chip detection method. Differences in CYP2C19 genotypes and metabolic phenotypes between the groups were compared. To analyze the efficacy of CYP2C19 metabolic phenotypes in discriminating between cerebral infarction and coronary artery disease, multiple logistic regression analysis was conducted after adjusting for gender, age, smoking history, drinking history, hypertension, and diabetes. Results: There were significant differences in the distribution of CYP2C19 genotypes and metabolic phenotypes between CI and CAD patients. The results of multivariate logistic regression (adjusted for sex, age, smoking, drinking, hypertension, and diabetes) indicated that CYP2C19 IM phenotype (IM vs EM: OR 1.443, 95% CI: 1.086-1.918, P=0.011) and CYP2C19 IM+PM phenotype (IM or PM vs EM: OR 1.440, 95% CI: 1.100-1.885, P=0.008) may be indicators of CI from CAD. Conclusion: CYP2C19 EM metabolic phenotype was dominant in CAD patients, and CYP2C19 IM metabolic phenotype was dominant in CI patients. After adjusting for other confounding factors, patients with the CYP2C19 IM metabolic phenotype were more likely to develop CI than CAD.

Lattice Compressive Strain of Co3O4 Induced by Synthetic Solvents Promotes Efficient Oxidation of Benzene at Low Temperature.

A series of Co3O4 with different surface defective structures were prepared by the solvothermal method and tested for the activity of benzene oxidation. The characterizations revealed that the synthetic solvent had a dramatic effect on the composition of Co3O4 precursors as well as the physicochemical properties of Co3O4. Although all Co3O4 exhibited a cubic spinel structure, Co3O4 prepared with triethylene glycol (Co-TEG) had the highest compressive strain due to the nature of high viscosity of triethylene glycol. These in turn affected the surface chemical structure and the low-temperature redox properties. Co-TEG exhibited the best benzene oxidation activity and showed excellent stability and good water resistance. In situ diffuse reflectance infrared Fourier transform spectroscopy was used to study the oxidation process of benzene. It was found that Co-TEG with more defective structures had abundant surface adsorbed oxygen and active lattice oxygen, which promoted the conversion of benzene and the corresponding intermediates at low temperature.

Right-to-left ventricle ratio determined by machine learning algorithms on CT pulmonary angiography images predicts prolonged ICU length of stay in operated chronic thromboembolic pulmonary hypertension.

OBJECTIVE: Right-to-left ventricle diameter ratio (dRV/dLV) on CT pulmonary angiography (CTPA) is a predictor of outcomes in non-operated chronic thromboembolic pulmonary hypertension (CTEPH) patients. The purpose of this study is to evaluate the performance of a novel machine learning (ML) algorithm for dRV/dLV measurement in operated CTEPH patients and its association with post-operative outcomes. METHODS: This retrospective study reviewed consecutive CTEPH patients who underwent pulmonary endarterectomy between 2013 and 2017. ML calculated dRV/dLV on pre-operative CTPA and compared with manual measures. Associations of dRV/dLV with patient characteristics and post-operative outcomes were evaluated including intensive care (ICU) and hospital length of stay (LOS) using multivariable linear regression analysis. Prolonged LOS was defined as greater than median. RESULTS: ML segmented the ventricles in 99/125 (79%) patients. The most common cause of failure was misidentification of the moderator band as the interventricular septum (7.9%). Mean dRV/dLV by ML was 1.4 ± 0.4 and strongly correlated with manual measures (r = 0.9-0.96 p < 0.0001). dRV/dLV was moderately correlated with measures of pulmonary hypertension on right heart catheterization and RV dilatation on echocardiogram (r = 0.5-0.6, p < 0.0001). dRV/dLV ≥ 1.2 was associated with proximal Jamieson type disease (p = 0.032), longer cardiopulmonary bypass (p = 0.037), aortic cross-clamp (p = 0.022) and circulatory arrest (p < 0.001) at surgery and dRV/dLV ≥ 1.6 with post-operative ECMO (p = 0.006). dRV/dLV was independently associated with prolonged ICU LOS (OR = 3.79, 95% CI 1.1-13.06, p = 0.035). CONCLUSION: dRV/dLV was associated with CTEPH severity and independently associated with prolonged ICU LOS. This CT parameter may therefore assist in perioperative planning. Further refinement of the ML algorithm or CTPA technique is required to avoid errors in ventricular segmentation. ADVANCES IN KNOWLEDGE: Automated right-to-left ventricle ratio measurement by machine learning is feasible and is independently associated with outcome after pulmonary endarterectomy.

A novel methylated cell-free DNA marker panel to monitor treatment response in metastatic prostate cancer.

Aim: This study examined circulating cell-free DNA (cfDNA) biomarkers associated with androgen treatment resistance in metastatic castration resistance prostate cancer (mCRPC). Materials & methods: We designed a panel of nine candidate cfDNA methylation markers using droplet digital PCR (Methyl-ddPCR) and assessed methylation levels in sequentially collected cfDNA samples from patients with mCRPC. Results: Increased cfDNA methylation in eight out of nine markers during androgen-targeted treatment correlated with a faster time to clinical progression. Cox proportional hazards modeling and logistic regression analysis further confirmed that higher cfDNA methylation during treatment was significantly associated with clinical progression. Conclusion: Overall, our findings have revealed a novel methylated cfDNA marker panel that could aid in the clinical management of metastatic prostate cancer.

Identification of specific clinical risk factors associated with the malignant transformation of oral epithelial dysplasia.

BACKGROUND: Factors that increase the risk of malignant transformation of oral epithelial dysplasia (OED) are not completely elucidated. METHODS: A retrospective chart review was performed assessing risk factors for transformation of OED, and cancer staging for transformed cases at Sunnybrook Health Sciences Centre. RESULTS: Two-hundred four patients were diagnosed with OED, and 16.7% (34) underwent malignant transformation. Risk factors associated with transformation included: heavy tobacco smoking, excessive EtOH consumption, non-homogenous leukoplakia, size >200 mm2 , moderate dysplasia or greater than moderate, progression of dysplasia grades, and immunosuppression. Transformed cases followed for a dysplastic lesion were associated with a stage-I cancer diagnosis, and cancer cases with no prior biopsy were associated with a stage-IV diagnosis. CONCLUSIONS: In addition to commonly cited risk factors, immunosuppression was associated with malignant transformation, including the use of topical steroids. Analyzing risk factors can help clinicians define risk of progression in patients with OED.

Evaluation of serum exosomal LncRNA-based biomarker panel for diagnosis and recurrence prediction of bladder cancer.

Exosomes are small membrane vesicles released by many cells. These vesicles can mediate cellular communications by transmitting active molecules including long non-coding RNAs (lncRNAs). In this study, our aim was to identify a panel of lncRNAs in serum exosomes for the diagnosis and recurrence prediction of bladder cancer (BC). The expressions of 11 candidate lncRNAs in exosome were investigated in training set (n = 200) and an independent validation set (n = 320) via quantitative real-time PCR. A three-lncRNA panel (PCAT-1, UBC1 and SNHG16) was finally identified by multivariate logistic regression model to provide high diagnostic accuracy for BC with an area under the receiver-operating characteristic curve (AUC) of 0.857 and 0.826 in training set and validation set, respectively, which was significantly higher than that of urine cytology. The corresponding AUCs of this panel for patients with Ta, T1 and T2-T4 were 0.760, 0.827 and 0.878, respectively. In addition, Kaplan-Meier analysis showed that non-muscle-invasive BC (NMIBC) patients with high UBC1 expression had significantly lower recurrence-free survival (P = 0.01). Multivariate Cox analysis demonstrated that UBC1 was independently associated with tumour recurrence of NMIBC (P = 0.018). Our study suggested that lncRNAs in serum exosomes may serve as considerable diagnostic and prognostic biomarkers of BC.

Application of Weighted Gene Co-expression Network Analysis for Data from Paired Design.

Investigating how genes jointly affect complex human diseases is important, yet challenging. The network approach (e.g., weighted gene co-expression network analysis (WGCNA)) is a powerful tool. However, genomic data usually contain substantial batch effects, which could mask true genomic signals. Paired design is a powerful tool that can reduce batch effects. However, it is currently unclear how to appropriately apply WGCNA to genomic data from paired design. In this paper, we modified the current WGCNA pipeline to analyse high-throughput genomic data from paired design. We illustrated the modified WGCNA pipeline by analysing the miRNA dataset provided by Shiah et al. (2014), which contains forty oral squamous cell carcinoma (OSCC) specimens and their matched non-tumourous epithelial counterparts. OSCC is the sixth most common cancer worldwide. The modified WGCNA pipeline identified two sets of novel miRNAs associated with OSCC, in addition to the existing miRNAs reported by Shiah et al. (2014). Thus, this work will be of great interest to readers of various scientific disciplines, in particular, genetic and genomic scientists as well as medical scientists working on cancer.

Biological effects of toosendanin, a triterpenoid extracted from Chinese traditional medicine.

Toosendanin (TSN) is a triterpenoid extracted from Melia toosendan Sieb et Zucc, which was used as a digestive tract-parasiticide and agricultural insecticide in ancient China. TSN was demonstrated to be a selective presynaptic blocker and an effective antibotulismic agent. By interfering with neurotransmitter release through an initial facilitation followed by a subsequent depression, TSN eventually blocks synaptic transmission at both the neuro-muscular junction and central synapses. Despite sharing some similar actions with botulinum neurotoxin (BoNT), TSN has a marked antibotulismic effect in vivo and in vitro. Studies suggest that the antibotulismic effect of TSN is achieved by preventing BoNT from approaching its enzymatic substrate, the SNARE protein. It is also found that TSN can induce differentiation and apoptosis in several cell lines, and suppress proliferation of various human cancer cells. TSN inhibits various K(+)-channels, selectively facilitates Ca(2+)-influx via L-type Ca(2+) channels and increases intracellular Ca(2+) concentration ([Ca(2+)](i)). The TSN-induced [Ca(2+)](i) increase and overload could be responsible for the TSN-induced biphasic effect on transmitter release, cell differentiation, apoptosis as well as the cytoxicity of TSN.

[Biological effects of toosendanin, an active ingredient of herbal vermifuge in Chinese traditional medicine].

The fact that the fruit and bark of plant belonging to family Melia could be used as digestive tract-parasiticide and agricultural insecticide was recorded about two thousand years ago in ancient China. Toosendanin (TSN, C30H38O11, FW=574), a triterpenoid derivative, was extracted from the bark of Melia toosendan Sieb. et Zucc. by Chinese scientists in 1950os and used as an ascarifuge in China instead of imported sendanin. Studies have demonstrated that TSN possesses special biological actions as well as considerable various values in scientific research, clinic medicine and agriculture. The first is that by interfering with neurotransmitter release by causing an initial facilitation, TSN eventually blocks synaptic transmission at both the neuromuscular junction and central synapses. The action might result from TSN-induced Ca(2+)-sensitivity change and final elimination of transmitter release machinery. The second is that despite sharing many similar actions with botulinum neurotoxin (BoNT) on blocking neuromuscular transmission, TSN has a markedly antibotulismic action in vivo and in vitro: TSN-treatment saves the botulism mice or monkeys from death; TSN-incubation in vitro or TSN-injection in vivo endows neuromuscular junction with a high tolerance to BoNT. Studies suggest that the antibotulismic action is achieved by preventing BoNT from approaching its enzymatic substrate, SNARE protein. The third, in recent years, it is also observed that TSN can induce differentiation and apoptosis in several cell lines, and suppress proliferation of various human cancer cells. The TSN-induced differentiation is Ca(2+)-dependent and the mitochondria-dependent apoptosis pathway is involved in the TSN-induced apoptosis. The fourth is that TSN inhibits various K(+) channels and selectively facilitates Ca(2+) current through L-type Ca(2+) channels and hence elevates [Ca(2+)](i). The TSN-induced [Ca(2+)](i) increase and overload could be responsible for the TSN-induced biphasic effect on neurotransmitter release, cell differentiation, apoptosis as well as the cytotoxicity of TSN.

Toosendanin interferes with pore formation of botulinum toxin type A in PC12 cell membrane.

AIM: Botulinum neurotoxins (BoNT) abort the process of neurotransmitter release at presynaptic motor nerve terminals, causing muscle paralysis. The ability of botulinum toxin to produce its effect is dependent on the ability of the light chain to cleave the SNARE proteins associated with transmitter release. Translocation of the light chain protease through the heavy chain-formed channel is a pivotal step in the intoxication process. Toosendanin (TSN), a triterpenoid derivative extracted from a Chinese traditional medicine, has been demonstrated to be an effective cure for experimental botulism. This study was designed to explore the antibotulismic mechanisms of toosendanin. METHODS: The inside-out single-channel recording patch-clamp technique was used to record the BoNT/A-induced currents in the presence and absence of TSN. RESULTS: Channel formation was delayed and the sizes of the channels were reduced in the TSN-treated PC12 cell membrane. CONCLUSION: The antibotulismic effect of TSN might occur via interference with toxin translocation.

Growth inhibition and apoptosis-induced effect on human cancer cells of toosendanin, a triterpenoid derivative from chinese traditional medicine.

Toosendanin, a triterpenoid derivative isolated from the barks of Melia toosendan Sieb et Zucc, has been used as an anthelmintic vermifuge against ascaris for more than fifty years in China. In the present study, we investigated the growth inhibition and apoptosis-induced effect of toosendanin on human cancer cells. The result showed that toosendanin significantly suppressed the proliferation of tested human cancer cell lines. The IC(50) values were less than 1.7 x 10(-7) M and U937 was the most sensitive cell line with a IC(50) of 5.4 x 10(-9) M. Flow cytometric analysis revealed that treatment of U937 cells with toosendanin resulted in a dose- and time-dependent accumulation of cells in the S phase with a concomitant decrease in cells processing to G(0)/G(1) phase. The growth inhibition of U937 cells after exposure to toosendanin was subsequently associated with the induction of apoptosis, as evidence by the typical condensed and fragmented nuclei, DNA fragmentation, and exposure of phosphatidylserine on the outer leaflet of plasma membrane. All these results indicated that toosendanin could serve as a potential candidate for anticancer drug.

The long-term effect of toosendanin on current through nifedipine-sensitive Ca2+ channels in NG108-15 cells.

Toosendanin is a triterpenoid derivative extracted from Melia toosendan Sieb et Zucc. Previous studies demonstrated that toosendanin could block neurotransmission and stimulate PC12 cell into differentiation and apoptosis. These actions of toosendanin were suggested to result from a continuous increase in Ca2+ influx, which led to intracellular Ca2+ overload. Here, we observed the long-term effect of toosendanin on Ca2+ channels in NG108-15 cells by whole-cell patch-clamp recording. Obtained data showed that a prolonged exposure to toosendanin induced a continuous increase in the Ca2+ influx in a concentration and time-dependent manner while a brief treatment induced an irreversible increase in Ca2+ influx in differentiated NG108-15 cells. The nifedipine-sensitive L-type currents were significantly increased after exposure to TSN while the nifedipine-resistant or omega-conotoxin MVIIC-sensitive currents were not affected.

Involvement of cytochrome c release and caspase activation in toosendanin-induced PC12 cell apoptosis.

Our previous study showed that toosendanin, a triterpenoid derivative isolated from a Chinese traditional medicine, could induce apoptosis in PC12 cells. In this study we confirmed the apoptosis-inducing effect of toosendanin in PC12 cells with new evidences in morphology and biochemistry: the shrinkage of cytosol, the condensation and fragmentation of nuclei and the formation of DNA ladder. It was also demonstrated that toosendanin decreased the PC12 cell viability in a time- and concentration-dependent manner. To elucidate the pathway linked with the toosendanin-induced apoptosis, the cytochrome c in the cytosol and the cleavage of poly(ADP-ribose) polymerase (PARP) were examined. The obtained results showed that toosendanin caused the release of cytochrome c from mitochondria into the cytosol and then led to the activation of caspase, indicating that the cytochrome c release and caspase activation were involved in the toosendanin-induced apoptosis process. These results suggested the possibility that toosendanin could serve as a candidate for anti-cancer drug.

Toosendanin increases free-Ca(2+) concentration in NG108-15 cells via L-type Ca(2+) channels.

AIM: To examine if toosendanin (TSN) affects intracellular free-Ca(2+) concentration ([Ca(2+)](i)) in neuroblastoma pluglioma hybrid cells (NG108-15 cells). METHODS: The [Ca(2+)](i) was determined by laser-scanning confocal microscopic imaging technique in which Fluo-3 was used as Ca(2+) indicator. RESULTS: TSN induced an increase in resting [Ca(2+)](i) and in high K(+)-evoked Ca(2+) transient in differentiated NG108-15 cells. The TSN-induced increase in [Ca(2+)](i) was dose-dependent and disappeared in CdCl(2-), nifedipine-containing or Ca(2+)-free solution, and appeared after washing out the Ca(2+) channel blockers or adding Ca(2+). CONCLUSION: TSN increased [Ca(2+)](i) in differentiated NG108-15 cells. The [Ca(2+)](i) enhancement was due to the influx of extracellular Ca(2+) and related to L-type Ca(2+) channels.

Toosendanin, a triterpenoid derivative, increases Ca2+ current in NG108-15 cells via L-type channels.

Toosendanin, a triterpenoid derivative extracted from Melia toosendan Sieb et Zucc, was demonstrated to be a selective presynaptic blocker and an effective antibotulismic agent in previous studies. Here, we observed its effects on Ca(2+) channels in NG108-15 cells by whole-cell patch-clamp recording. Obtained data showed that toosendanin concentration dependently increased the high-voltage-activated (HVA) Ca(2+) current with an EC(50) of 5.13 microM in differentiated NG108-15 cells. The enhancement effect was still observed when the cells were pretreated with 5 microM omega-conotoxin MVIIC. However, when the cells were preincubated with 5 microM nifedipine or 10 microM verapamil-containing solution, the effect was absent. In undifferentiated NG108-15 cells, which only express T-type Ca(2+) channels, toosendanin did not affect Ca(2+) currents. These results show that toosendanin increases Ca(2+) influx in NG108-15 cells via L-type Ca(2+) channels.

Effects of demethylzeylasteral and celastrol on spermatogenic cell Ca2+ channels and progesterone-induced sperm acrosome reaction.

The male antifertility effect of a water-chloroform extract of Tripterygium wilfordii Hook. f. (GTW) and several monomers isolated from GTW has attracted worldwide interest. In the present study, the effects of two isolated monomers from GTW, demethylzeylasteral and celastrol, on the Ca(2+) channels in mouse spermatogenic cells and on the sperm acrosome reaction were investigated by whole-cell patch-clamp recording and chlortetracycline staining methods, respectively. The results showed that demethylzeylasteral concentration-dependently and in a partially reversible manner inhibited the Ca(2+) current in spermatogenic cells with an IC(50) of 8.8 microg/ml. Celastrol decreased the Ca(2+) current in the cells time-dependently and irreversibly. The changes in the activation and inactivation time constants of Ca(2+) currents after application of these two compounds were also examined. Demethylzeylasteral increased both activation and inactivation time constants of Ca(2+) currents, and celastrol had no significant effect on them. The two compounds also inhibited significantly the sperm acrosome reaction initiated by progesterone. These data suggest that inhibition of Ca(2+) currents could be responsible for the antifertility activity of these compounds.