An Overview of Drug Substance Manufacturing Processes.

To develop a comprehensive understanding of pharmaceutical drug substance manufacturing (DSM) processes, we conducted a data mining study to examine 50 new drug applications (NDAs) approved in 2010-2016. We analyzed the prevalence of several frequently deployed in-process control (IPC) techniques and postreaction workup procedures, as well as the operational conditions specified for reactions and workups. Our findings show that crystallization and high-performance liquid chromatography (HPLC) were the most commonly used workup steps and in-process controls, respectively, in drug substance manufacturing. On average, each NDA implemented 12.6 in-process controls and 11.3 workups. Operation time for reactions and workup procedures varied from a few minutes to multiple days, though 61% of these were between 1 and 10 h.

Tumor Growth Assessment by Computed Tomography Perfusion Imaging (CTPI), Perfusion-Weighted Imaging (PWI), and Diffusion-Weighted Imaging (DWI) in a Rabbit Pleural Squamous Cell Carcinoma VX2-Implanted Model.

BACKGROUND Computed tomography perfusion imaging (CTPI) and perfusion-weighted imaging (PWI) are non-invasive technologies that can quantify tumor vascularity and blood flow. This study explored the blood flow information, tumor cell viability, and hydrothoraces in a rabbit pleural VX2-implanted model through use of CTPI, PWI, and DWI. MATERIAL AND METHODS A pleural VX2-implanted model was established in 58 New Zealand white rabbits. CTPI, PWI, and DWI were applied with a 16-slice spiral CT and an Archival 1.5 T dual-gradient MRI. RESULTS Compared with muscle tissue, PV, PEI, and BV of parietal and visceral pleural tumor implantation rabbits showed significant differences. The t values of PV, PEI, and BV between parietal and visceral pleura were 2.08, 2.29, and 2.88, respectively. Compared with muscle tissue, WIR, WOR, and MAXR of parietal and visceral pleural tumor implantation rabbits showed significant differences. In parietal pleural tumor implantation rabbits, the section surface of lesion tissues was 5.2±2.7 cm². Hydrothorax appeared 6.0±2.0 days after tumor implantation. The mean value of ADC was 1.5±0.6. In visceral pleural tumor implantation rabbits, the section surface of lesion tissues was 1.6±0.8 cm². Hydrothorax appeared 7.0±3.0 days after tumor implantation. The mean value of ADC was 1.4±0.5. The t values of the above 3 indices for the parietal and visceral pleura were 1.85, 1.83, and 1.76, respectively (P<0.05). CONCLUSIONS The combined application of CTPI, PWI, and DWI accurately and visually reflects the blood perfusion of tumor tissues and quantitatively analyzes blood flow information and the mechanism underlying hydrothorax generation in tumor tissues.

MAPK inhibitors modulate Smad2/3/4 complex cyto-nuclear translocation in myofibroblasts via Imp7/8 mediation.

Mitogen-activated protein kinase (MAPK) pathway-dependent linker phosphorylation of Smad2/3 and subsequent formation of Smad2/3/4 complex and its nuclear translocation are crucial for dysregulated transforming growth factor beta (TGF)-ß/Smad signaling in liver fibrosis. Abrogation of this critical step of TGF-ß/Smad signaling leading to liver fibrosis could provide new insights for future therapy, but the mechanisms remain incompletely understood. In pursuit, we investigated the subcellular expression and nuclear trafficking of the rate limiting Smad2/3/4 complex in exogenous TGF-ß1-stimulated myofibroblasts (MFBs) using three MAPK-specific inhibitors. Our results showed that exogenous TGF-ß1 stimulation of MFBs produced both increased protein expression and nuclear translocation of phosphorylated (p)-Smad2C/L, oncogenic pSmad3L, Smad4, importin7/8 (Imp7/8), and plasminogen activator inhibitor (PAI)-1 (Protein and mRNA), while decreased Smad7 protein expression. However, the MAPK-specific inhibitors differentially reversed these observations; for instance, ERK-specific inhibitor blocked the expression and nuclear translocation of pSmad2C/L, while both JNK and p38-specific inhibitors blocked the expression and nuclear translocation of pSmad2C/L and oncogenic pSmad3L. The MAPK-specific inhibitors had no significant effect on the total protein expression of Smad4, but rather significantly blocked its nuclear translocation. All the MAPK-specific inhibitors restored Smad7 expression and also decreased Imp7/8 and PAI-1 (Protein and mRNA) expression. Evidently, the MAPK-specific inhibitors blocked Smad2/3/4 complex formation via restoration of inhibitory Smad7 expression and blockade of Smad3L phosphorylation, while they blocked nuclear translocation of Smad2/3/4 complex through inhibition of Imp7/8 leading to decreased PAI-1 (Protein and mRNA) expression.

Formation of Organic Molecular Nanocrystals under Soft Confinement.

Methods to produce nano-sized organic molecular crystals in thin films are of great interest in the pharmaceutical industry due to the potential benefit of increased solubility of poorly soluble drugs and the advantages of film-based dosage forms over traditional tablet/capsule-based dosage form. One method to directly form organic nanocrystals is by crystallization in confined environments where the overall crystallization volume is constrained. We report the use of a novel solution impregnation method to form nanocrystals in polymer matrices with various microstructures in order to study the structure of the confined nanocrystals and the role of soft confinement and polymer chemistry on the nucleation process of nano-sized crystals. The particle diameter correlates with the microstructure of the polymer matrices and the nucleation kinetics. In addition, by carefully choosing the experimental conditions and the polymer matrix, polymorph control of nanocrystals can be achieved. Solid-state nuclear magnetic resonance (ssNMR) was used to examine the local structure of nanocrystals inside the polymer matrices and crystal polymer interactions. This method may serve as a novel formulation method to obtain nanocrystals of poorly soluble active pharmaceutical ingredients (APIs) for pharmaceutical industry.

Bone marrow stroma-mediated resistance to FLT3 inhibitors in FLT3-ITD AML is mediated by persistent activation of extracellular regulated kinase.

A consistent pattern of response has been observed when FMS-like tyrosine kinase 3 (FLT3) tyrosine kinase inhibitors (TKIs) have been used as monotherapy to treat patients with relapsed or refractory FLT3- internal tandem duplication (ITD) acute myeloid leukaemia (AML). Circulating blasts are cleared from the peripheral blood, while bone marrow blasts are either unaffected or are cleared from the marrow at a much slower rate. We used an in vitro model of FLT3-ITD AML blasts co-cultured with normal human bone marrow stromal cells to investigate the basis for this dichotomous response pattern to FLT3 inhibitors. We have found that in blasts on stroma, potent FLT3 inhibition predominantly results in cell cycle arrest rather than apoptosis. The anti-apoptotic effect is mediated through a combination of direct cell-cell contact and soluble factors. The addition of exogenous FLT3 ligand (FL) augments the protection, primarily by shifting the 50% inhibitory concentration for FLT3 inhibition upwards. Cytokine-activated extracellular regulated kinase (ERK), rather than STAT5, appears to be the most important downstream signalling protein mediating the protective effect, and inhibition of MEK significantly abrogates stromal-mediated resistance. These findings explain the phenomenon of peripheral blood versus bone marrow blast responses and suggest that the combination of potent FLT3 inhibition and MEK inhibition is a promising strategy for the treatment of FLT3-ITD AML.

Terminal myeloid differentiation in vivo is induced by FLT3 inhibition in FLT3/ITD AML.

A hallmark of cancer is the disruption of differentiation within tumor cells. Internal tandem duplication mutations of the FLT3 kinase (FLT3/ITD) occur commonly in acute myeloid leukemia (AML) and are associated with poor survival, leading to efforts to develop FLT3 kinase inhibitors. However, FLT3 inhibitors have thus far met with limited success, inducing only a clearance of peripheral blasts with minimal BM responses. Quizartinib is a novel potent and selective FLT3 inhibitor currently being studied in clinical trials. In 13 of 14 FLT3/ITD AML patients with normal karyotype treated with quizartinib, we observed terminal myeloid differentiation of BM blasts in association with a clinical differentiation syndrome. The single patient whose blasts failed to differentiate had a preexisting C/EBPα mutation and another developed a C/EBPα mutation at disease progression, suggesting a mechanism of resistance to FLT3 inhibition. In vitro, in primary blasts cocultured with human BM stroma, FLT3 inhibition with quizartinib induced cell-cycle arrest and differentiation rather than apoptosis. The present study is the first description of terminal differentiation of cancer cells in patients treated with a tyrosine kinase inhibitor. These data highlight the importance of the differentiation block in the patho-genesis of AML.

Non-destructive strategy to extract sustainable helix and high-strength Musa core fibers for rapid water conduction and evaporation.

The reuse and development of natural waste resources is a hotspots and challenges in the research of new fiber materials and the resolution of environmental concern globally. Herein, this study aimed to develop a simple and direct manual extraction process to extract Musa core fibers (MCFs) for rapid water conduction and evaporation. Through simple processes such as ring cutting and stretching, this green and non-destructive inside-out extraction strategy enabled Musa fibers to be naturally and harmlessly degummed from natural Musa stems, with good maintenance of the fiber structure and highly helical morphology. The extracted fibers are composed of regularly and closely arranged cellulose nanofibrils in the shape of ribbon spirally arranged multi-filaments, and the single filament is about 2.65 µm. The high-purity fibers exhibit ultra-high tensile strength under a non-destructive extraction process, and the ultimate tensile strength in dry state is as high as 742.95 MPa. The tensile strength is affected by the number of fiber bundles, which shows that tensile strength and tensile modulus is higher than those of vascular bundle fibers in dry or wet condition. In addition, the MCFs membrane indicates good water conductivity, with a water absorption height of 50 mm for the sample in only 60 s. Moreover, the water evaporation rate of MCFs reaches 1.37 kg m-2 h-1 in 30 min, which shows that MCFs have excellent water conductivity and evaporation rate compared with ordinary cotton fibers. These results indicate that MCFs have great potential in replacing the use of chemical methods to extract fibers from vascular bundles, providing an effective way to achieve sustainability in quick-drying applications, as well as in the sustainable development of natural waste resources.

Spontaneous thermal energy transfer and anti-gravitational water pumping using Al2O3 fiber-enhanced flexible nonwoven material as a high-performance and self-floating solar evaporator.

Solar-driven evaporation is promising to address water scarcity. However, preserving the heat inside evaporators instead of allowing run-off, and synergistically utilizing it to wick water from the bulk, is still underexplored. Herein, a dual-functional bridge of longitudinal orientated channels of Al2O3 fibers (AOFs) embedded in a multi-layered nonwoven evaporator was proposed to create a buffer for spontaneous thermal conduction and anti-gravitational water pumping. As a self-floating system with high porosity and flexibility, benefiting from the strong water transporting ability and high thermal conductivity of the AOFs, a superhigh evaporation rate (2.79 kg m-2 h-1 under 1 sun) can be achieved with great stability and durability. This work highlights the potential of promoting thermal management using a large-scale vapour chamber and mass-producible nonwoven technology to prepare a high-performance evaporator for practical applications.

FLT3 ligand impedes the efficacy of FLT3 inhibitors in vitro and in vivo.

We examined in vivo FLT3 inhibition in acute myeloid leukemia patients treated with chemotherapy followed by the FLT3 inhibitor lestaurtinib, comparing newly diagnosed acute myeloid leukemia patients with relapsed patients. Because we noted that in vivo FLT3 inhibition by lestaurtinib was less effective in the relapsed patients compared with the newly diagnosed patients, we investigated whether plasma FLT3 ligand (FL) levels could influence the efficacy of FLT3 inhibition in these patients. After intensive chemotherapy, FL levels rose to a mean of 488 pg/mL on day 15 of induction therapy for newly diagnosed patients, whereas they rose to a mean of 1148 pg/mL in the relapsed patients. FL levels rose even higher with successive courses of chemotherapy, to a mean of 3251 pg/mL after the fourth course. In vitro, exogenous FL at concentrations similar to those observed in patients mitigated FLT3 inhibition and cytotoxicity for each of 5 different FLT3 inhibitors (lestaurtinib, midostaurin, sorafenib, KW-2449, and AC220). The dramatic increase in FL level after chemotherapy represents a possible obstacle to inhibiting FLT3 in this clinical setting. These findings could have important implications regarding the design and outcome of trials of FLT3 inhibitors and furthermore suggest a rationale for targeting FL as a therapeutic strategy.

Synthesis, antimalarial activity and cytotoxic potential of new monocarbonyl analogues of curcumin.

A series of novel monocarbonyl analogues of curcumin have been designed, synthesized and tested for their activity against Molt4, HeLa, PC3, DU145 and KB cancer cell lines. Six of the analogues showed potent cytotoxicity towards these cell lines with IC(50) values below 1 µM, which is better than doxorubicin, a US FDA approved drug. Several analogues were also found to be active against both CQ-resistant (W2 clone) and CQ-sensitive (D6) strains of Plasmodium falciparum in an in-vitro antimalarial screening. This level of activity warrants further investigation of the compounds for development as anticancer and antimalarial agents.

[Comparison of left ventricular function evaluation using gated myocardial perfusion imaging with different software of quantitative analysis].

In the present study, the accuracy of the 4-dimensional model single photon emission computed tomography (4D-MSPECT) and quantitative gated single photon emission computed tomography (QGS) was investigated for assessing left ventricular end-diastolic (EDV), end-systolic volume (ESV) and ejection fraction (EF) from gated myocardial perfusion imaging (G-MPI) using left ventriculography (LVG) as reference. From December 2008 to June 2011, 85 patients, who underwent rest G-MPI and LVG (within 30 days) in West China Hospital, Sichuan University, were retrospectively recruited. EDV, ESV, and EF were calculated from G-MPI using 4D-MSPECT and QGS. Eighty-five patients (47 men, 38 women; age 57 +/- 13 years) were finally analyzed. Correlation between results of G-MPI and LVG was high for EDV, r = 0.89 (4D-MSPECT), r = 0.81 (QGS); ESV, r = 0.97 (4D-MSPECT), R = 0.95(QGS); EF, r = 0.95 (4D-MSPECT), r = 0.93 (QGS). 4D-MSPECT and QGS underestimated EDV significantly compared with LVG [(125 +/- 20) mL (4D-MSPECT), (118 +/- 39) mL (QGS), (131 +/- 33)mL (LVG)]. The ESV, 4D-MSPECT and QGS values did not differ significantly from LVG [(47 +/- 32) mL (4D-MSPECT), (53 +/- 29) mL (QGS), (49 +/- 37) mI (LVG)]. For LVEF, only QGS yielded values were significantly lower than LVG [61% +/- 21% (4D-MSPECT), 55% +/- 17% (QGS), and 63% +/- 19% (LVG)]. EDV, ESV, and EF as determined by 4D-MSPECT and QGS from G-MPI agree well with relevant values with LVG. However, Algorithm-inherent also showed slightly over- or under-estimation of volumes. Therefore, separated normal databases should be set up for each algorithm.

Clinical significance of the controlling nutritional status (CONUT) score in gastric cancer patients: A meta-analysis of 9,764 participants.

Background: The clinical value of the controlling nutritional status (CONUT) score has been widely reported in multiple malignancies. The aim of this study is to investigate the association between the CONUT score and clinical outcomes in patients with gastric cancer. Methods: A comprehensive literature search of electronic databases including PubMed, Embase, and Web of Science was performed up to December 2022. The primary endpoints were survival outcomes and postoperative complications. Subgroup analysis and sensitivity analysis were performed during the pooled analysis. Results: Nineteen studies including 9,764 patients were included. The pooled results indicated that patients in the high CONUT group had a worse overall survival (HR = 1.70 95%CI: 1.54-1.87; P < 0.0001; I 2 = 33%) and recurrence-free survival (HR = 1.57; 95%CI: 1.36-1.82; P < 0.0001; I 2 = 30%), and a higher risk of complications (OR = 1.96; 95%CI: 1.50-2.57; P < 0.0001; I 2 = 69%). In addition, a high CONUT score was significantly associated with larger tumor size, higher percentage of microvascular invasion, later TNM stage and fewer patients receiving adjuvant chemotherapy, but not with tumor differentiation. Conclusion: Based on existing evidence, the CONUT score could act as a valuable biomarker to predict clinical outcomes in patients with gastric cancer. Clinicians could use this useful indicator to stratify patients and formulate individual treatment plans.

Preparation of hydroxyapatite coated porous carbon nanofibres for DEX loading and enhancing differentiation of BMSCs.

The proliferation and differentiation of bone mesenchymal stem cells (BMSCs) in vitro are the key properties of bone tissue engineering for biomaterials. In this study, hydroxyapatite (HA) coated porous carbon nanofibres (PCNFs) were prepared to load dexamethasone (DEX) and further improve the differentiation ability of the BMSCs. Various characterisations were applied to reveal the DEX loading efficacy and biocompatibility, especially the differentiation strength. The results showed that HA could be successfully coated on the PCNFs by pretreating the surface using PEG conjugation. With an increase of HA, the particle diameter increased and the DEX loading decreased. In vitro experiments proved higher cell viability, alkaline phosphatase (ALP) activity, calcium nodule secretion ability and the RUNX2 protein expression, indicating that the as-prepared was of great biocompatibility and optimised osteoconductivity, which was attributed to the componential imitation to natural bone and the accelerated BMSCs differentiation. Consequently, the novel DEX loaded and HA coated PCNFs can provide potential applications in bone tissue regeneration.

Stabilization of a premixed CH4/O2/N2 flame using femtosecond laser-induced plasma.

The influence of femtosecond laser-induced plasma (FLIP) on the stability of a premixed CH(4)/O(2)/N(2) flame is investigated at atmospheric pressure. The laser energy, laser repetition rate, the equivalence ratios, and the volume percentage of oxygen in O(2)/N(2) blends are varied. Our findings indicate that the flame blow-off velocity is a function of these parameters. It has been experimentally found that the flame blow-off velocity increases by a factor of two with FLIP than without FLIP. A high-repetition-rate and a great energy laser-induced plasma flameholding, as a non-intrusive optical flameholding, may be a feasible alternative for any combustor.

[Influence of gated myocardial perfusion imaging with different acquisition models on the assessment of left ventricular function].

This paper is aimed to investigate the influence of gated myocardial perfusion imaging (G-MPI) with 8- and 16-frame acquisition models on the assessment of left ventricular function. Patients prepared for stress and rest G-MPI were prospectively recruited from January 2010 to January 2011 in the Department of Nuclear Medicine of West China Hospital, Sichuan University. Two separate G-MPI studies, one with 8 and the other with 16 frames, were simultaneously acquired during a single gantry orbit using Concurrent Imaging technique. We calculated the left ventricular ejection fraction (EF) and volumes using the Auto Quant software. Forty-eight patients (29 men, 19 women; average age 51 +/- 16 years old) were finally analyzed. The differences in left ventricular EF between 8- and 16-frame were small: 3.27% (95% CI: 6.41%-0.12%) for post-stress and 3.13% (95% CI: 5.93%-0.32%) for rest. Both using 8 and 16 frames, there were significantly larger volumes and lower EF in patients with stress-induced ischemia than without. As for detecting left ventricular EF, 8-frame and 16-frame acquisition models should not be mutually alternated.

Prognostic value of pan-immune-inflammation value in colorectal cancer patients: A systematic review and meta-analysis.

Background: The pan-immune-inflammation value (PIV) has been reported as a novel prognostic biomarker in multiple malignancies. The aim of this study is to investigate the prognostic value of the PIV in patients with colorectal cancer. Methods: We comprehensively searched electronic databases including PubMed, Embase and Web of Science up to August 2022. The endpoints were survival outcomes. Hazard ratios (HRs) with 95% confidence intervals (CIs) for survival data were collected for analysis. Results: Six studies including 1879 participants were included. A significant heterogeneity in the PIV cut-off value among studies was observed. The combined results indicated that patients in the high baseline PIV group had a worse overall survival (HR=2.09; 95%CI: 1.67-2.61; P<0.0001; I2 = 7%) and progression-free survival (HR=1.82; 95%CI: 1.49-2.22; P<0.0001; I2 = 15%). In addition, early PIV increase after treatment initiation was significantly associated with decreased overall survival (HR=1.79; 95%CI: 1.13-2.93; P=0.01; I2 = 26%), and a trend toward poor progression-free survival (HR=2.00; 95%CI: 0.90-4.41; P=0.09; I2 = 70%). Conclusion: Based on existing evidence, the PIV could act as a valuable prognostic index in patients with colorectal cancer. However, the heterogeneity in the PIV cut-off value among studies should be considered when interpreting these findings.

Plasma-assisted combustion of methane using a femtosecond laser.

We first research the effects of femtosecond-laser-induced plasma (FLIP) on a laminar premixed methane/oxygen/nitrogen flame speed with a wide range of equivalence ratios (0.8-2.0) at atmospheric pressure. It is experimentally found that the flame speed increases by 30.8% at equivalence ratio 1.33, and the effects of the FLIP on the flame speed are more remarkable when the methane is rich. The self-emission spectra from the flame and the plasma are studied, and the presence of the oxygen atom is likely to be a key factor in enhancing flame speed.

Chemical validation of phosphodiesterase C as a chemotherapeutic target in Trypanosoma cruzi, the etiological agent of Chagas' disease.

Trypanosoma cruzi phosphodiesterase (PDE) C (TcrPDEC), a novel and rather unusual PDE in which, unlike all other class I PDEs, the catalytic domain is localized in the middle of the polypeptide chain, is able to hydrolyze cyclic GMP (cGMP), although it prefers cyclic AMP (cAMP), and has a FYVE-type domain in its N-terminal region (S. Kunz et al., FEBS J. 272:6412-6422, 2005). TcrPDEC shows homology to the mammalian PDE4 family members. PDE4 inhibitors are currently under development for the treatment of inflammatory diseases, such as asthma, chronic pulmonary diseases, and psoriasis, and for treating depression and serving as cognitive enhancers. We therefore tested a number of compounds originally synthesized as potential PDE4 inhibitors on T. cruzi amastigote growth, and we obtained several useful hits. We then conducted homology modeling of T. cruzi PDEC and identified other compounds as potential inhibitors through virtual screening. Testing of these compounds against amastigote growth and recombinant TcrPDEC activity resulted in several potent inhibitors. The most-potent inhibitors were found to increase the cellular concentration of cAMP. Preincubation of cells in the presence of one of these compounds stimulated volume recovery after hyposmotic stress, in agreement with their TcrPDEC inhibitory activity in vitro, providing chemical validation of this target. The compounds found could be useful tools in the study of osmoregulation in T. cruzi. In addition, their further optimization could result in the development of new drugs against Chagas' disease and other trypanosomiases.

An automated modular assembly line for drugs in a miniaturized plant.

We report here a fully automated, end-to-end, integrated continuous manufacturing process for a small-molecule generic medication with built-in quality assurance. The entire process fits into a box of 30.7 m2 modular footprint and a total residence time of less than 30 h, with a throughput up to 40.3 × 106 tablets per year.

Acceleration of osteogenic differentiation of preosteoblastic cells by chitosan containing nanofibrous scaffolds.

To accelerate bone tissue formation, scaffolds better provide stimulatory niches to the residing cells for osteogenic differentiation. In this study, polycaprolactone (PCL) nanofibers containing various amounts of chitosan (0, 3, 9, and 23%) were prepared and evaluated for their osteogenic differentiation of preosteoblasts in 2D and 3D cultures. Characterization of the obtained nanofibers revealed that average fiber diameter, hydrophilicity, Young's modulus, and fiber degradation were closely correlated with the amount of chitosan in PCL nanofibers. Incorporation of chitosan in PCL nanofibers not only improved the adhesion and proliferation of MC 3T3-E1 cells but also elevated calcium deposition, alkaline phosphatase (ALP) activity, and the expression of osteopontin (OPN) compared to PCL alone nanofibers. Culture of cell-rich 3D constructs prepared by layer-by-layer assembling MC 3T3-E1 with chitosan containing PCL nanofibers led to a uniform tissue formation with significant mineralization at 21 days. In all, chitosan containing PCL nanofibers are superior to PCL nanofibers in promoting bone tissue formation.