Sustainability-Driven Accelerated Shear-Mediated Immunoassay for Amyotrophic Lateral Sclerosis Detection.

Healthcare facilities produce millions of tons of waste annually, with a significant portion consisting of diagnostic plasticware. Here, we introduce a new detection platform that completely replaces traditional assay plates with a piece of membrane, offering a much greener and more sustainable alternative. The membrane, integrated within the portable vortex fluidic device (P-VFD), enables rapid detection of a clinically relevant protein biomarker, urinary p75ECD. This biomarker is utilized to evaluate the prognosis, disease severity, and progression of amyotrophic lateral sclerosis (ALS). This assay has a limit-of-detection (LOD) of 4.03 pg, which is comparable to the plate-based assay (2.24 pg) and has been optimized through a full factorial design of experiments (DOE). P-VFD has great potential in quantifying p75ECD in human biofluids and can significantly reduce the assay time to 5 min compared to the current plate-based p75ECD ELISA assay (3 days), with at least a 4.4-fold reduction in the usage of the detection antibody.

Joint MAPLE: Accelerated joint T1 and T 2 * $$ {{\mathrm{T}}_2}

PURPOSE: Quantitative MRI finds important applications in clinical and research studies. However, it is encoding intensive and may suffer from prohibitively long scan times. Accelerated MR parameter mapping techniques have been developed to help address these challenges. Here, an accelerated joint T1, T 2 * $$ {{\mathrm{T}}_2}^{\ast } $$ , frequency and proton density mapping technique with scan-specific self-supervised network reconstruction is proposed to synergistically combine parallel imaging, model-based, and deep learning approaches to speed up parameter mapping. METHODS: Proposed framework, Joint MAPLE, includes parallel imaging, signal modeling, and data consistency blocks which are optimized jointly in a combined loss function. A scan-specific self-supervised reconstruction is embedded into the framework, which takes advantage of multi-contrast data from a multi-echo, multi-flip angle, gradient echo acquisition. RESULTS: In comparison with parallel reconstruction techniques powered by low-rank methods, emerging scan specific networks, and model-based T 2 * $$ {{\mathrm{T}}_2}^{\ast } $$ estimation approaches, the proposed framework reduces the reconstruction error in parameter maps by approximately two-fold on average at acceleration rates as high as R = 16 with uniform sampling. It can outperform evaluated parallel reconstruction techniques up to four-fold on average in the presence of challenging sub-sampling masks. It is observed that Joint MAPLE performs well at extreme acceleration rates of R = 25 and R = 36 with error values less than 20%. CONCLUSION: Joint MAPLE enables higher fidelity parameter estimation at high acceleration rates by synergistically combining parallel imaging and model-based parameter mapping and exploiting multi-echo, multi-flip angle datasets. Utilizing a scan-specific self-supervised reconstruction obviates the need for large data sets for training while improving the parameter estimation ability.

Thermo-hydraulic performance optimization of a disk-shaped microchannel heat sink applying computational fluid dynamics, artificial neural network, and response surface methodology.

The current research focuses on optimizing the Nusselt number (Nu) and pressure drop (ΔP) in a bionic fractal heat sink. The artificial neural network (ANN) and response surface methodology (RSM) were used to model the thermos-hydraulic behavior of the MCHS. The aspect ratios of t/b (cavities' upper side to bottom side ratio) and h/b (cavities' height to bottom side ratio), as well as the Reynolds number, were set as the independent variables in both ANN and RSM models. After finding the optimum state for the copper-made MCHS (containing the optimum design of the cavities along with the best applied velocity), different materials were tested and compared with the base case (heat sink made of copper). The obtained results indicated that both ANN and RSM models (with determination coefficient of 99.9 %) could exactly anticipate heat transfer and ΔP to a large extent. To achieve the optimal design of the microchannel heat sink (MCHS) with the objective of maximizing Nu and minimizing ΔP, the efficiency index of the device was evaluated. The analysis revealed that the highest efficiency index (1.070 by RSM and 1.067 by ANN methods) was attained when the aspect ratios were t/b = 0.2, h/b = 0.2, and the Reynolds number was 1000. Next, the effect of the different materials on heat sink performance was investigated, and it was observed that by reducing the thermal conductivity, the thermal resistance of the heat sink increased and its overall performance decreased.

MiR-21 and Tocilizumab interactions improve COVID-19 myocarditis outcomes.

BACKGROUND: Myocarditis is now one of the most fatal and morbid complications of COVID-19. Many scientists have recently concentrated on this problem. OBJECTIVES: This study assessed the effects of Remdesivir (RMS) and Tocilizumab (TCZ) in COVID-19 myocarditis. DESIGN: Observational, cohort study. METHODS: Patients with COVID-19 myocarditis were enrolled in the study and divided into three groups, TCZ-treated, RMS-treated, and Dexamethasone-treated patients. After 7 days of treatment, patients were reassessed for improvement. RESULTS: TCZ significantly improved patients' ejection fraction in 7 days, but it had limited efficacy. RMS improved inflammatory characteristics of the disease, but RMS-treated patients showed exacerbated cardiac function over 7 days, and the mortality rate with RMS was higher than TCZ. TCZ protects the heart by decreasing the miR-21 expression rate. CONCLUSION: Using Tocilizumab in early diagnosed COVID-19 myocarditis patients can save their cardiac function after hospitalization and decrease the mortality rate. miR-21 level determines the outcome and responsiveness of COVID-19 myocarditis to treatment.

Printable Hydrogel Arrays for Portable and High-Throughput Shear-Mediated Assays.

Hydrogels have been widely used to entrap biomolecules for various biocatalytic reactions. However, solute diffusion in these matrices to initiate such reactions can be a very slow process. Conventional mixing remains a challenge as it can cause irreversible distortion or fragmentation of the hydrogel itself. To overcome the diffusion-limit, a shear-stress-mediated platform named the portable vortex-fluidic device (P-VFD) is developed. P-VFD is a portable platform which consists of two main components, (i) a plasma oxazoline-coated polyvinyl chloride (POx-PVC) film with polyacrylamide and alginate (PAAm/Alg-Ca2+) tough hydrogel covalently bound to its surface and (ii) a reactor tube (L × D: 90 mm × 20 mm) where the aforementioned POx-PVC film could be readily inserted for reactions. Through a spotting machine, the PAAm/Alg-Ca2+ hydrogel can be readily printed on a POx-PVC film in an array pattern and up to 25.4 J/m2 adhesion energy can be achieved. The hydrogel arrays on the film not only offer a strong matrix for entrapping biomolecules such as streptavidin-horseradish peroxidase but are also shear stress-tolerant in the reactor tube, enabling a >6-fold increase in its reaction rate after adding tetramethylbenzidine, relative to incubation. Through using the tough hydrogel and its stably bonded substrate, this portable platform effectively overcomes the diffusion-limit and achieves fast assay detection without causing appreciable hydrogel array deformation or dislocation on the substrate film.

Satralizumab, Novel Interleukine-6 Inhibitor for Preventing Descending Thoracic Aorta Aneurysm Development.

BACKGROUND: Descending thoracic aorta aneurysm (dTAA) has increasing incidence and, if left untreated, could lead to death. There is not any study of satralizumab treatment for preventing dTAA formation and progression. MATERIALS AND METHODS: Forty male 10-week-old Rattus norvegicus were enrolled in the experiment. They were divided into four equal groups: dTAA treated with saline (dTAA-P) and dTAA treated with satralizumab (dTAA-S). One of the control groups was treated with saline (C-P), and the other was treated with satralizumab (C-S). Satralizumab and saline were used once every 2 weeks, subcutaneously 120 mg for 4 weeks. dTA diameter was measured at days 0, 3, 7, 14, 21, and 28. RESULTS: IL-6 level was measured on the 7th day that showed significantly increased IL-6 serum level in dTAA-P rats compared to C-P. Maximal dTA diameter (%MAD) was obtained at day 14, which was scientifically matched to the aorta aneurysm definition (>50% increase in diameter). From the seventh day, a significant difference in %MAD was observed between dTAA-P and dTAA-S groups. However, the %MAD of these two groups was significantly higher than control groups till the end of the 28th day. CONCLUSION: Using an IL-6 inhibitor agent to prevent dTAA formation and progression showed promising results. It suggests that using the IL-6 inhibitors in susceptible persons can be considered a lifesaving therapeutic approach.

Advanced Glycation End Product Blocker Drugs Have a Great Potential to Prevent Diabetic Cardiomyopathy in an Animal Model of Diabetes Mellitus Type-2.

Introduction: Sphingosine 1 phosphate (S1P) is a product of the sphingosine kinase 1 (SphK1) enzyme. Increased S1P can lead to tissue fibrosis that is also one of the pathways for developing diabetic cardiomyopathy. Advanced glycation end products (AGEs) increase S1P in cells. The study is aimed at using aminoguanidine (AG) as an AGEs blocker drug to prevent diabetic cardiomyopathy. Materials and methods. 210 rats were enrolled in the study. Diabetes mellitus type-2 was induced, and rats were divided into AG treated diabetic and nondiabetic groups. The heart histology was assessed with Masson's trichrome and hematoxylin-eosin staining. Cardiac function was measured with transthoracic echocardiography. S1P level and SphK1 gene expression were measured by western-blot and RT-qPCR, respectively. Results: Results showed that S1P level increases in diabetes, and its augmentation in cardiac tissue with K6PC-5 leads to cardiac fibrosis. 50 and 200 mg/kg of AG prevented cardiac fibrosis, but 100 mg/kg had no significant preventive effect. AG suppressed the SphK1 gene expression and reduced the fibrotic effect of S1P. AG preserved cardiac function by keeping ejection fraction and fractional shortening within the normal range in diabetic rats. Conclusion: AG has a suppressor effect on SphK1 gene expression besides its AGEs blocker role. AG is a potential drug to use in diabetic patients for preventing the development of diabetic cardiomyopathy. Other drugs that have AGEs or S1P blocker effects are a good choice for diabetic cardiomyopathy prevention.

The utility of SYNTAX score predictability by electrocardiogram parameters in patients with unstable angina.

BACKGROUND: SYNTAX score is one of the risk assessment systems to predict cardiac events in acute coronary syndrome patients. Despite the large number of SYNTAX score benefits, invasive methods such as coronary angiography are necessary to perform the scoring. We hypothesized that ECG parameters could predict the SYNTAX score in unstable angina patients. METHODS: During the retrospective cohort study, a total number of 876 patients were diagnosed with unstable angina. After applying the exclusion criteria, 600 patients were divided into tertiles based on the SYNTAX scores as low (0-22), intermediate (23-32), and high (≥ 33). The association between ECG parameters and SYNTAX score was investigated. RESULTS: The study included 65% men and 35% women with a mean age of 62.4 ± 9.97 years. The delayed transition zone of QRS complex, ST-depression in inferior-lateral territories or/and in all three territories, and T-wave inversion in lateral territory were significant (p < 0.05) independent predictors of intermediate SYNTAX score. High SYNTAX score was predicted by the presence of prolonged P wave duration, ST-depression in lateral territory or/and anterior-lateral territories, ST-elevation in aVR-III leads or/and aVR-III-V1 leads. Among those, all three territories ST-depression (AUC: 0.611, sensitivity: 75%, specificity: 51%) and aVR + III ST-elevation (AUC: 0.672, sensitivity: 50.12%, specificity: 80.50%) were the most accurate parameters to predict intermediate and high SYNTAX scores, respectively. CONCLUSION: The present study demonstrates that accompanying the STE in the right side leads (aVR, III, V1) with ST-depression in other leads indicates the patients with high SYNTAX score; meanwhile, diffuse ST-depression without ST-elevation is a marker for intermediate SYNTAX score in unstable angina patients and can be applied for early risk stratification and intervention.

Removal of methylene blue via bioinspired catecholamine/starch superadsorbent and the efficiency prediction by response surface methodology and artificial neural network-particle swarm optimization.

This paper demonstrates coupling of the artificial neural network (ANN) technique with the particle swarm optimization (PSO) method and compares the performance of ANN-PSO with response surface methodology (RSM) in prediction of the adsorption of methylene blue (MB) by a novel bio-superadsorbent. To this, a starch-based superadsorbent was synthesized using acrylic acid and acryl amid polymers and then catecholamine functional groups were combined onto the surface with oxidative polymerization of dopamine. The adsorption of MB was considered as a function of pH, dye concentration, and contact time. The best topology of the ANN was found to be 3-7-1, and prediction model of the adsorption capacity was demonstrated as a matrix of explicit equations. ANN-PSO is more accurate than RSM. The results revealed that the root-mean-square error, correlation coefficient, and normalized standard deviation for the ANN-PSO are 22.46, 0.99, and 16.83, respectively, while for RSM are 82.89, 0.98, and 65.41, respectively.

What is the Role of Preoperative Breathing Exercises in Reducing Postoperative Atelectasis after CABG?

BACKGROUND: Atelectasis and hypoxemia are frequently reported after coronary artery bypass graft surgery (CABG). Some studies confirm the benefits of breathing exercises on pulmonary complications, but the efficacy of preoperative breathing exercises in patients undergoing CABG is controversial. In this study, the effect of preoperative breathing exercises on the incidence of atelectasis and hypoxemia in patients candidate for CABG was examined. METHODS: In a single-blinded randomized clinical trial, 100 patients who were undergoing coronary artery bypass graft surgery were randomly allocated into two groups of experimental and control, each consisted of 50 patients. Before the operation, experimental group patients were enrolled in a protocol including deep breathing, cough and incentive spirometer. In the control group, hospital routine physiotherapy was implemented. All the patients received the hospital routine physiotherapy once a day for 2 to 3 minutes in the first four days postoperatively. Arterial blood gases and atelectasis were compared between groups. RESULTS: There was no significant difference between groups in terms of atelectasis and hypoxemia (p Value>0.05). CONCLUSION: Preoperative breathing exercise does not reduce pulmonary complications in patients undergoing CABG.

Effect of weight losing on the clinical status improvement of patients with knee osteoarthritis.

OBJECTIVE: Osteoarthritis causes severe pain and disability in joints, one of the most prevalent involved joints is the knee joint. There are several therapeutics ways to control pain and disability, but almost none of them are definite treatment. In this article, we tried to reveal the effect of weight loss on improving symptoms of knee osteoarthritis as an effective and permanent therapeutic approach. METHODS: We chose 62 patients with grade 1-2 (mild to moderate) knee osteoarthritis and divided them equally into case and control groups. Patients should not had used NSAIDs at least for 6 months before study initiation. Symptoms severity was measured by WOMAC and VAS questionnaires before and after 3 months follow up. Weight and BMI were recorded too. Case group was suggested to have weight loss diet of less fat and carbohydrates and control group did not have any limitation. RESULTS: Comparison of variables' average of case and control groups was not logistically meaningful at the initiation and after the end of the study. But there was a meaningful correlation between variables' changes and lifestyle change in both groups, especially in WOMAC and VAS scores. All variables in case group had statistically meaningful differences between their amounts at the beginning and after the end of the study, on the contrary of the control group. CONCLUSION: In the comparison of our study with similar studies in the world. We deduced that weight loss can improve symptoms of knee osteoarthritis even in short time weight loss diet (3 months). TRIAL REGISTRATION NUMBER: ZUMS.REC.1394.94.

Bioinspired catecholamine/starch composites as superadsorbent for the environmental remediation.

Focusing on the encouraging properties of starch-based composite materials, starch­g­(acrylic acid­co­acrylamide) superabsorbent was synthesized using solution polymerization method, and then the catecholamine functional groups were introduced on to pore surface of the absorbent via oxidative polymerization of dopamine (DA). The adsorbent was optimized in terms of the monomers' mass ratio and synthesis conditions, and characterized by different characterization techniques. The polydopamine (PDA) coating thickness was estimated using transmission electron microscopy (TEM) image and it was found to be 83 nm. The bimodal mesoporous adsorbent with 5914.66% swelling ratio bearing micropores with a specific surface area of 2.8031 m2 g-1 was used for the adsorption of methylene blue (MB) as a model water pollutant dye. The maximum adsorption capacity was obtained 2276 mg g-1 at pH 9 and within 100 min. The adsorbent with unprecedented super high adsorption capacity can be encouraging from different environmental remediation points of view.

Cytotoxic effect of 2, 5-dimethyl-celecoxib as a structural analog of celecoxib on human colorectal cancer (HT-29) cell line.

Dimethyl-celecoxib (DMC), a close derivative of celecoxib (CXB) with a low COX-2 inhibitory function, exhibits significant anti-neoplastic properties. In this study, we have investigated the effect of CXB and DMC on the human HT-29 cell line. The cellular viability, caspase-3 activity, and VEGF, NF-κB, and COX-2 genes expressions were assessed respectively with MTT, colorimetric, and real-time RT-PCR methods. DMC, a close analogue of CXB, was more potent in inhibiting the growth of cells (IC50: 23.45 µM at 24 hr) than CXB (IC50: 30.41 µM at 24 hr). Both CXB and DMC caused a significant difference in caspase-3 activity compared to the control group. DMC significantly decreased the NF-κB expression. Down-regulation of the COX-2 mRNA expression in the celecoxib-treated group was significant compared with that in the DMC-treated group. Alterations in the mRNA expression of VEGF were not significant between the groups. Owing to the more potent growth inhibitory effects of DMC compared to that of celecoxib, it may be important to conduct research on the anticancer application of this compound, which can reduce the side effects relating to COX2 inhibition.

Quantitative Proteomic Analysis of Cellular Responses to a Designed Amino Acid Feed in a Monoclonal Antibody Producing Chinese Hamster Ovary Cell Line

Background: Chinese hamster ovary (CHO) cell line is considered as the most common cell line in the biopharmaceutical industry because of its capability in performing efficient post-translational modifications and producing the recombinant proteins, which are similar to natural human proteins. The optimization of the upstream process via different feed strategies has a great impact on the target molecule expression and yield. Methods: To determine and understand the molecular events beneath the feed effects on the CHO cell, a label-free quantitative proteomic analysis was applied. The proteome changes followed by the addition of a designed amino acid feed to the monoclonal antibody producing CHO cell line culture medium were investigated. Results: The glutathione synthesis, the negative regulation of the programmed cell death, proteasomal catabolic process, and the endosomal transport pathway were up-regulated in the group fed with a designed amino acid feed compared to the control group. Conclusion: Our findings could be helpful to identify new targets for metabolic engineering.

Comparison of fenofibrate and pioglitazone effects on patients with nonalcoholic fatty liver disease.

INTRODUCTION: Nonalcoholic fatty liver disease (NAFLD) is known to be a health-related problem; there is no proven treatment for NAFLD. However, a wide range of possible therapies have been proposed and studied. In the current study, we aimed to compare the therapeutic effects of fenofibrate and pioglitazone on NAFLD. PATIENTS AND METHODS: In this randomized clinical trial study (ethic number: ZUMS.REC.1393.133), patients with NAFLD and alanine aminotransferase in range of 1-1.5 folds of normal and BMI (25-35) were studied. Blood lipids and liver enzymes were measured. The patients were divided randomly into three groups (recipient of fenofibrate, pioglitazone, and exercise). After the patients completed the course of treatment, liver enzymes were measured. RESULTS: According to the results of this study, 90 patients with NAFLD were divided into three groups of 30 patients. All variables at the beginning of the study showed no significant difference among the three groups, but after the treatment period, the results showed that the levels of alanine aminotransferase, aspartate transaminase, systolic blood pressure, diastolic blood pressure, and BMI changed significantly: the levels decreased in all groups. CONCLUSION: This study showed beneficial effects of fenofibrate and pioglitazone in patients with fatty liver. Further studies with larger study populations on the effects of these drugs on fatty liver, lipid profile, blood glucose, and insulin are suggested.

Imatinib and its combination with 2,5-dimethyl-celecoxibinduces apoptosis of human HT-29 colorectal cancer cells.

Mono-targeting by imatinib as a main antitumor agent does not always accomplish complete cancer suppression. 2,5-dimethyl-celecoxib (DMC) is a close structural analog of the selective cyclooxygenase-2 (COX-2) inhibitor, celecoxib, that lacks COX-2 inhibitory function. In this study, we aimed to show the apoptotic effects of imatinib in combination with DMC in human HT-29 colorectal cancer (CRC) cells. HT-29 CRC cells were treated with IC50 dose of imatinib (6.60 µM), DMC (23.45 µM), and their combination (half dose of IC50) for 24 h. The caspase-3 activity was estimated with colorimetric kit. The caspase-3 gene expression was evaluated by real-time PCR method. There was a significant up-regulation in caspase-3 enzyme activity and caspase-3 expression by imatinib and its half dose combination with DMC as compared to control. As a summary, the results of this study strongly suggest that half dose combination of imatinib with DMC induced apoptosis as potent as full dose imatinib in human HT-29 CRC cells, while minimizing undesired side effects related to imatinib mono-therapy. This study also pointed towards possible caspase-dependent actions of imatinib and DMC.

Designed Amino Acid Feed in Improvement of Production and Quality Targets of a Therapeutic Monoclonal Antibody.

Cell culture feeds optimization is a critical step in process development of pharmaceutical recombinant protein production. Amino acids are the basic supplements of mammalian cell culture feeds with known effect on their growth promotion and productivity. In this study, we reported the implementation of the Plackett-Burman (PB) multifactorial design to screen the effects of amino acids on the growth promotion and productivity of a Chinese hamster ovary DG-44 (CHO-DG44) cell line producing bevacizumab. After this screening, the amino acid combinations were optimized by the response surface methodology (RSM) to determine the most effective concentration in feeds. Through this strategy, the final monoclonal antibody (mAb) titre was enhanced by 70%, compared to the control group. For this particular cell line, aspartic acid, glutamic acid, arginine and glycine had the highest positive effects on the final mAb titre. Simultaneously, the impact of the designed amino acid feed on some critical quality attributes of bevacizumab was examined in the group with highest productivity. The product was analysed for N-glycan profiles, charge variant distribution, and low molecular weight forms. The results showed that the target product quality has been improved using this feeding strategy. It was shown how this strategy could significantly diminish the time and number of experiments in identifying the most effective amino acids and related concentrations in target product enhancement. This model could be successfully applied to other components of culture media and feeds.

Implications of mechanism-based inhibition of CYP2D6 for the pharmacokinetics and toxicity of MDMA.

The aim of this study was to model the in vivo kinetic consequences of mechanism-based inhibition (MBI) of CYP2D6 by 3,4 methylenedioxymethamphetamine (MDMA, ecstasy). A model with physiologically-based components of drug metabolism was developed, taking account of change in the hepatic content of active CYP2D6 due to MBI by MDMA. Based on the in vitro information, plasma concentration time profiles of MDMA after various doses were computed and compared with reported observations. The analysis suggested that a typical recreational MDMA dose could inactivate most hepatic CYP2D6 within an hour, and the return to a basal level of CYP2D6 could take at least 10 days. Thus, the genetic polymorphism of CYP2D6 and coadministration of CYP2D6 inhibitors may have less impact on MDMA pharmacokinetics and the risk of acute toxicity than previously thought. This is consistent with clinical observations that indicate no obvious link between inherited CYP2D6 deficiency and acute MDMA intoxication.

The impact of experimental design on assessing mechanism-based inactivation of CYP2D6 by MDMA (Ecstasy).

MDMA (3-4-methylenedioxymethamphetamine, commonly known as Ecstasy) is a potent mechanism-based inhibitor (MBI) of cytochrome P450 2D6 (CYP2D6), causing quasi-irreversible inhibition of the enzyme in vitro. An evaluation of the in vivo implications of this phenomenon depends on the accuracy of the estimates of the parameters that define the inhibition in vitro, namely k(inact) (the maximal inhibition rate) and KI (the inactivation constant). These values are determined in two steps, pre-incubation of the enzyme with the inhibitor (enzyme inactivation), followed by dilution and further incubation to measure residual enzyme activity with a probe substrate. The aim of this study was to assess the impact of different dilutions and probe substrate concentrations on the estimates of k(inact) and KI using recombinantly expressed CYP2D6. Enzyme activity was measured by the conversion of dextromethorphan (DEX) to dextrorphan (DOR). Dilution factors of 1.25, 2, 5, 10, 25 and 50 (DEX at 30 microM) gave mean (+/-SE) values of k(inact) (min-1) of 0.20+/-0.06, 0.21+/-0.05, 0.31+/-0.06, 0.37+/-0.11, 0.51+/-0.10 and 0.58+/-0.08, respectively, and KI (microM) values (after correction for non-specific microsomal binding) of 2.22+/-1.90, 2.80+/-1.34, 5.78+/-2.07, 6.36+/-2.93, 3.99+/-1.57 and 4.86+/-1.37, respectively. Accordingly, high (e.g. 50 fold) and low (e.g. 1.25 fold) dilutions were associated with statistically significant differences in kinetic values (p <0.05). Varying DEX concentration (10-100 microM) was not associated with significant changes in k(inact) and KI values when a five-fold dilution was used (with the exception of a lower KI at 10 microM DEX). High dilution was also shown to reduce non-specific microsomal binding of MDMA. The changes in the two kinetic parameters were dependent on the experimental procedure and shown to be unlikely to have a material influence on the maximum inhibition of CYP2D6 expected in vivo after typical recreational doses of MDMA (50-100 mg), since the potency of inhibition was high. The different values of the kinetic parameters were predicted to have a marginal influence on the time for recovery of enzyme activity following re-synthesis of CYP2D6.