In silico study of alkaloids with quercetin nucleus for inhibition of SARS-CoV-2 protease and receptor cell protease.

Covid-19 disease caused by the deadly SARS-CoV-2 virus is a serious and threatening global health issue declared by the WHO as an epidemic. Researchers are studying the design and discovery of drugs to inhibit the SARS-CoV-2 virus due to its high mortality rate. The main Covid-19 virus protease (Mpro) and human transmembrane protease, serine 2 (TMPRSS2) are attractive targets for the study of antiviral drugs against SARS-2 coronavirus. Increasing consumption of herbal medicines in the community and a serious approach to these drugs have increased the demand for effective herbal substances. Alkaloids are one of the most important active ingredients in medicinal plants that have wide applications in the pharmaceutical industry. In this study, seven alkaloid ligands with Quercetin nucleus for the inhibition of Mpro and TMPRSS2 were studied using computational drug design including molecular docking and molecular dynamics simulation (MD). Auto Dock software was used to evaluate molecular binding energy. Three ligands with the most negative docking score were selected to be entered into the MD simulation procedure. To evaluate the protein conformational changes induced by tested ligands and calculate the binding energy between the ligands and target proteins, GROMACS software based on AMBER03 force field was used. The MD results showed that Phyllospadine and Dracocephin-A form stable complexes with Mpro and TMPRSS2. Prolinalin-A indicated an acceptable inhibitory effect on Mpro, whereas it resulted in some structural instability of TMPRSS2. The total binding energies between three ligands, Prolinalin-A, Phyllospadine and Dracocephin-A and two proteins MPro and TMRPSS2 are (-111.235 ± 15.877, - 75.422 ± 11.140), (-107.033 ± 9.072, -84.939 ± 10.155) and (-102.941 ± 9.477, - 92.451 ± 10.539), respectively. Since the binding energies are at a minimum, this indicates confirmation of the proper binding of the ligands to the proteins. Regardless of some Prolinalin-A-induced TMPRSS2 conformational changes, it may properly bind to TMPRSS2 binding site due to its acceptable binding energy. Therefore, these three ligands can be promising candidates for the development of drugs to treat infections caused by the SARS-CoV-2 virus.

The genetic basis of early-onset hereditary ataxia in Iran: results of a national registry of a heterogeneous population.

BACKGROUND: To investigate the genetics of early-onset progressive cerebellar ataxia in Iran, we conducted a study at the Children's Medical Center (CMC), the primary referral center for pediatric disorders in the country, over a three-year period from 2019 to 2022. In this report, we provide the initial findings from the national registry. METHODS: We selected all early-onset patients with an autosomal recessive mode of inheritance to assess their phenotype, paraclinical tests, and genotypes. The clinical data encompassed clinical features, the Scale for the Assessment and Rating of Ataxia (SARA) scores, Magnetic Resonance Imaging (MRI) results, Electrodiagnostic exams (EDX), and biomarker features. Our genetic investigations included single-gene testing, Whole Exome Sequencing (WES), and Whole Genome Sequencing (WGS). RESULTS: Our study enrolled 162 patients from various geographic regions of our country. Among our subpopulations, we identified known and novel pathogenic variants in 42 genes in 97 families. The overall genetic diagnostic rate was 59.9%. Notably, we observed PLA2G6, ATM, SACS, and SCA variants in 19, 14, 12, and 10 families, respectively. Remarkably, more than 59% of the cases were attributed to pathogenic variants in these genes. CONCLUSIONS: Iran, being at the crossroad of the Middle East, exhibits a highly diverse genetic etiology for autosomal recessive hereditary ataxia. In light of this heterogeneity, the development of preventive strategies and targeted molecular therapeutics becomes crucial. A national guideline for the diagnosis and management of patients with these conditions could significantly aid in advancing healthcare approaches and improving patient outcomes.

TPP1 Variants in Iranian patients: A Novel Pathogenic Homozygous Variant Causing Neuronal Ceroid Lipofuscinosis 2.

Introduction: TPP1 variants have been identified as a causative agent of neuronal ceroid lipofuscinosis 2 disease, that ataxia is one of its clinical features. Therefore, here, molecular study of TPP1 variants is presented in an Iranian cohort and a novel pathogenic variant is described. Methods: This investigation was conducted as a cross-sectional study in a tertiary referral hospital, Children's Medical Center, Pediatrics Center of Excellence. Clinical presentations and pedigrees were documented. Patients with cerebellar ataxia were enrolled in this study. Next-generation sequencing was applied to confirm the diagnosis. Segregation and bioinformatics analyses were also done for the variants using Sanger sequencing. Results: Forty-five patients were included in our study. The mean age of onset was 104 (+55.60) months (minimum = 31 months, maximum = 216 months). The majority of cases (73.3%) were born to consanguineous parents and only 1 patient (2.2%) had an affected sibling. Of the 45 patients, only 1 patient with a novel pathogenic variant (c.1425_1425+1delinsAT, p.A476Cfs*15) in the TPP1 gene was identified. Discussion: The main strength of current study is the relatively large sample size. Besides, a novel pathogenic variant could be important toward the diagnosis and management of this condition. With significant advances in various therapies, early diagnosis could improve the treatments using personalized-based medicine.

Application of magnetic dispersive solid phase extraction combined with solidification of floating organic droplet-based dispersive liquid-liquid microextraction and GC-MS in the extraction and determination of polycyclic aromatic hydrocarbons in honey.

A magnetic dispersive solid phase extraction method combined with solidification of floating organic droplet-based dispersive liquid-liquid microextraction has been validated for the extraction of polycyclic aromatic hydrocarbons from honey samples. For this purpose, a carbonised cellulose-ferromagnetic nanocomposite was used as a sorbent through the magnetic dispersive solid phase extraction. For preparation of the sorbent, first, carbonised cellulose nanoparticles were created by treating cellulose filter paper with concentrated solution of sulfuric acid. Then, the prepared nanoparticles were loaded onto Fe3O4 nanoparticles through coprecipitation. In the extraction process, first, a few mg of the sorbent was added to the diluted honey solution and dispersed in it using vortex agitation. The particles were then separated and the adsorbed analytes were eluted with an organic solvent. The eluent was taken and after mixing with a water-immiscible extraction solvent was used in the following solidification of floating organic droplet-based dispersive liquid-liquid microextraction procedure. By performing the extraction process under the obtained optimum conditions, low limits of detection (0.08-0.17 ng g-1) and quantification (0.27-0.57 ng g-1), satisfactory precision (relative standard deviations ≤ 5.0%), and wide linear range (0.57-500 ng g-1) with great coefficients of determination (r2≥ 0.9986) were obtained.

Surfactant-enhanced air-agitation liquid-liquid microextraction of polycyclic aromatic hydrocarbons from edible oil using magnetic deep eutectic solvent prior to HPLC determination.

Herein, an air-agitation liquid-liquid microextraction procedure was developed for the extraction of several polycyclic aromatic hydrocarbons from edible oil samples. In this study, the extraction procedure was achieved using a new magnetic deep eutectic solvent as the extraction solvent, in which there was no need for centrifugation. To enhance the rate of extraction of the analytes from the samples, the method was promoted by the use of surfactant addition. The extracted analytes were determined by high-performance liquid chromatography with a diode array detector. The influence of various parameters on the extraction efficiency was studied by response surface methodology using a central composite design. Under optimal conditions, linear calibration curves for the target analytes were achieved in the range of 0.43-250 ng g-1. The limits of detection and quantification were in the ranges of 0.04-0.13 and 0.13-0.43 ng g-1, respectively. The repeatability of the method in terms of intra- and inter-day precision was ≤4.7% and ≤6.7%, respectively. The extraction recovery of the method ranged from 75 to 88%. The obtained results show that the proposed method is efficient for the analysis of the target analytes in various oil samples without obvious matrix effects. Pyrene was found in olive oil at a concentration of 42 ng g-1.

Selective extraction of apixaban from plasma by dispersive solid-phase microextraction using magnetic metal organic framework combined with molecularly imprinted polymer nanocomposite.

This research aims to synthesize a specific and efficient sorbent to use in the extraction of apixaban from human plasma samples and its determination by high-performance liquid chromatography-tandem mass spectrometry. High specific surface area of metal-organic framework, magnetic property of iron oxide nanoparticles, selectively of molecular imprinted polymer toward the analyte, and the combination of dispersive solid-phase extraction method with a sensitive analysis system provided an efficient analytical method. In this study, first, a molecularly imprinted polymer combined with magnetic metal organic framework nanocomposite was prepared and then characterized using different techniques. Then the sorbent particles were used for selective extraction of the analyte from plasma samples. The efficiency of the method was improved by optimizing effective parameters. According to the validation results, wide linear range (1.02-200 ng mL-1 ), acceptable coefficient of determination (0.9938), low limit of detection (0.32 ng mL-1 ) and limit of quantification (1.02 ng mL-1 ), high extraction recovery (78%), and good precision (relative standard deviations ≤ 2.9% for intra- (n = 6) and interday (n = 6) precisions) were obtainable using the proposed method. These outcomes showed the high potential of the proposed method for screening apixaban in the human plasma samples.

Histopathologic evaluation of the inflammatory factors and stromal cells in the endometriosis lesions: A case-control study.

Background: Endometriosis is a multifaceted gynecological disorder defined as a benign estrogen-dependent chronic inflammatory process in which endometrial glands and stroma-like tissues are located outside the uterine cavity. It affects around 2-10% of all women during their reproductive years. Objective: This study aimed to evaluate the traffic of mesenchymal stem cells and inflammatory factors toward the lesions. Materials and Methods: Ten samples of normal endometrium and eutopic endometrium were studied as a control group and 10 ectopic samples were considered as a case group. Hematoxylin and eosin staining was used to evaluate stromal cells and inflammatory cells. Immunohistochemical staining was performed to show the presence of proliferating cell nuclear antigen in the lesions. The cells were digested and cultured in the laboratory to study cell proliferation. The number of cells and vessels were counted with Image J software, and data analysis was performed with Prism software. Results: Data analysis showed that the number of stromal cells and vessels in ectopic tissue were significantly higher than the control group (p < 0.001). Also, the number of inflammatory cells, including neutrophils, monocytes, lymphocytes, and macrophages, in the ectopic group was much higher than in the control group (p < 0.005). Conclusion: By expanding the number of blood vessels, blood flow increases, and cell migration to tissues is facilitated. The accumulation of inflammatory cells, especially macrophages, stimulates the growth of stem cells and helps implant cells by creating an inflammatory process.

Magnetic dispersive solid-phase extraction of some polycyclic aromatic hydrocarbons from honey samples using iron (III) oxinate nanocomposite as an efficient sorbent.

In this work, iron (III) oxinate magnetic nanocomposite was synthesized and employed as an efficient sorbent for the magnetic dispersive solid-phase extraction of some polycyclic aromatic hydrocarbons from honey samples. In the following, dispersive liquid-liquid microextraction procedure was used for further preconcentration of the analytes. The prepared sorbent was characterized using Fourier transform infrared spectrophotometry, X-ray diffractometry, vibrating sample magnetometry, energy dispersive X-ray spectroscopy, and scanning electron microscopy. The results verified the successful formation of the magnetic sorbent. In the extraction process, the sorbent was added into an aqueous solution and the mixture was vortexed. After completing the adsorption process, the supernatant phase was separated in the presence of a magnet and the analytes adsorbed onto sorbent were eluted by acetonitrile. Then, microliter-level 1,1,1-trichloroethane was mixed with the obtained acetonitrile and injected into NaCl solution. Finally, one microliter of the sedimented phase was injected into gas chromatography-flame ionization detector after centrifugation. Under the optimum conditions, a great repeatability (relative standard deviation equal or less than 5 and 6% for intra- and interday precisions, respectively), acceptable extraction recoveries (59-84%), high enrichment factors (118-168), and low limits of detection and quantification (0.16-0.36 and 0.56-1.22 ng/g, respectively) were acquired.

Dispersive micro-solid-phase extraction of aflatoxins from commercial soy milk samples using a green vitamin-based metal-organic framework as an efficient sorbent followed by high performance liquid chromatography-tandem mass spectrometry determination.

In the current work, for the first time, a vitamin-based metal-organic framework constructed from cobalt ions and vitamin B3 has been used as a sorbent in dispersive micro-solid-phase extraction. The proposed method was used to extract and enrich aflatoxins (B1, B2, G1, and G2) from soy milk samples before their quantification by high performance liquid chromatography-tandem mass spectrometry. In this work, first the metal-organic framework was synthesized and characterized using techniques such as X-ray diffraction, Fourier transform infrared spectrophotometry, scanning electron microscopy, nitrogen adsorption/desorption, and energy-dispersive X-ray spectroscopy. Then it was used as an efficient sorbent in the proposed dispersive micro-solid-phase extraction. For this purpose, after precipitating the proteins of soy milk sample with the aid of trichloroacetic acid, the supernatant phase was taken, mixed with the synthesized sorbent, and vortexed. After centrifuging, the analytes loaded on the adsorbent were eluted with methanol to transfer them into an organic phase which was compatible with the subsequently employed separation system. The adsorption capacity of the synthesized MOF for aflatoxins B1, B2, G1, and G2 were 0.77, 0.83, 0.70, and 0.54 mg g-1, respectively. Under the best experimental situations, satisfactory outcomes including acceptable extraction recoveries (64-75%), low limits of detection (11.3-48.2 ng L-1) and quantification (42.8-161.6 ng L-1), and good repeatability (relative standard deviations equal or less than 4.0 and 4.7% for intra- and inter- day precisions, respectively) were obtained. In addition, green synthesis of the metal-organic framework (using vitamin B3 as a linker, water as the reaction solvent, and mild conditions) and usage low amount or volume of the adsorbent and organic solvents during the extraction process were the other beneficial aspects of this work which caused the suggested analytical method to be environmentally friendly.

Sensitive determination of vincristine in plasma of children with leukaemia using vortex-assisted dispersive liquid-liquid microextraction based on hydrophobic deep eutectic solvent.

Vincristine has a wide spectrum of clinical activity and is currently used in the treatment of leukemia. Despite its high therapeutic properties, vincristine has common side effects. Accordingly, it is desirable to determine vincristine in plasma for the use of the drug with strict monitoring. In the present research, for the first time a hydrophobic deep eutectic solvent (DES) composed of methyltrioctylammonium chloride (MTOAC) and n-butanol in a molar ratio of 1 : 3 was used as the extractant in dispersive liquid-liquid microextraction (DLLME) for the extraction and determination of vincristine in the plasma of children with leukemia prior to its analysis by high-performance liquid chromatography-ultraviolet detection (HPLC-UV). Under optimal experimental conditions, the method showed good linearity with a correlation coefficient (R 2) of 0.9986 in the linear range of 0.06-300 µg L-1, low limit of detection of 0.02 µg L-1 and acceptable extraction efficiency (EE) of 88.4%. In the final stage of the study, this proposed technique was successfully applied to determine vincristine in real plasma, and the obtained results demonstrated the ability of the synthesized DES to extract drugs from biological fluids.

Application of calcium oxide as an efficient phase separation agent in temperature-induced counter-current homogenous liquid-liquid extraction of aflatoxins from dried fruit chips followed by high-performance liquid chromatography-tandem mass spectrometry determination.

A temperature-induced counter-current homogenous liquid-liquid extraction procedure performed in a burette has been proposed for the isolation of aflatoxins B1, B2, G1, and G2 from various fruit chip samples. In this method, a homogenous solution of deionized water and cyclohexylamine is added to the solid sample and the resulted mixture is vortexed. In the following, the liquid phase is taken and passed through the burette filled with a mixture of calcium oxide (as a phase separation agent) and sand (to avoid clumping the calcium oxide). By doing so, the temperature of the solution is increased by hydration of calcium oxide and consequently, the homogenous state is broken and the aflatoxins are migrated into the resulted tiny droplets of cyclohexylamine. This phase is collected on the top of the solution owing to its low density with respect to an aqueous solution. Numerous parameters which can affect the efficiency of the suggested approach were evaluated and under the best situations, great repeatability, low limits of determination and quantification, and high extraction recoveries were acquired. In the end, the suggested approach was employed for the quantification of the selected aflatoxins in various fruit chips samples marketed in Tabriz City, Iran.

The safety and efficacy of umbilical cord blood mononuclear cells in individuals with spastic cerebral palsy: a randomized double-blind sham-controlled clinical trial.

INTRODUCTION: The current multi-center, randomized, double-blind study was conducted among children with cerebral palsy (CP) to assess the safety and efficacy of umbilical cord blood mononuclear cell (UCB-MNC). We performed the diffusion tensor imaging to assess the changes in the white matter structure. METHODS: Males and females aged 4 to 14 years old with spastic CP were included. Eligible participants were allocated in 4:1 ratio to be in the experimental or control groups; respectively. Individuals who were assigned in UCB-MNC group were tested for human leukocyte antigen (HLA) and fully-matched individuals were treated with UCB-MNCs. A single dose (5 × 106 /kg) UCB-MNCs were administered via intrathecal route in experimental group. The changes in gross motor function measure (GMFM)-66 from baseline to one year after treatment were the primary endpoints. The mean changes in modified Ashworth scale (MAS), pediatric evaluation of disability inventory (PEDI), and CP quality of life (CP-QoL) were also evaluated and compared between groups. The mean changes in fractional anisotropy (FA) and mean diffusivity (MD) of corticospinal tract (CST) and posterior thalamic radiation (PTR) were the secondary endpoints. Adverse events were safety endpoint. RESULTS: There were 72 included individuals (36 cases in each group). The mean GMFM-66 scores increased in experimental group; compared to baseline (+ 9.62; 95%CI: 6.75, 12.49) and control arm (ß: 7.10; 95%CI: 2.08, 12.76; Cohen's d: 0.62) and mean MAS reduced in individuals treated with UCB-MNCs compared to the baseline (-0.87; 95%CI: -1.2, -0.54) and control group (ß: -0.58; 95%CI: -1.18, -0.11; Cohen's d: 0.36). The mean PEDI scores and mean CP-QoL scores in two domains were higher in the experimental group compared to the control. The imaging data indicated that mean FA increased and MD decreased in participants of UCB-MNC group indicating improvements in white matter structure. Lower back pain, headaches, and irritability were the most common adverse events within 24 h of treatment that were related to lumbar puncture. No side effects were observed during follow-up. CONCLUSIONS: This trial showed that intrathecal injection of UCB-MNCs were safe and effective in children with CP. TRIAL REGISTRATION: The study was registered with ClinicalTrials.gov ( NCT03795974 ).

Improved magnetic solid-phase extraction based on magnetic sorbent obtained from sand for the extraction of pesticides from fruit juice.

BACKGROUND: A combination of magnetic solid-phase extraction using an efficient and cheap magnetic sorbent obtained from sand and dispersive liquid-liquid microextraction has been developed for the extraction of nine multiclass pesticides (clodinafop-propargyl, haloxyfop-R-methyl, fenoxaprop-P-ethyl, oxadiazon, penconazole, diniconazole, chlorpyrifos, fenazaquin, and fenpropathrin) from commercial fruit juices (sour cherry, pomegranate, grape, watermelon, orange, apricot, and peach juices). The enriched pesticides were determined by gas chromatography-flame ionization detector and gas chromatography-mass spectrometry. The sorbent was natural iron oxide entrapped in silica along with some impurities. In this method, to extract the analytes from the samples, an appropriate amount of the magnetic sorbent (at mg level) is added. Then the sorbent particles are isolated from the solution using an external magnetic field and the adsorbed analytes are desorbed from the sorbent by acetone. In the following, a dispersive liquid-liquid microextraction procedure is carried out to concentrate the analytes more and to reach low limits of detection. RESULTS: Under optimized extraction conditions, the method revealed satisfactory repeatability (relative standard deviation ≤8% for intra-day and inter-day precision), reasonable extraction recovery (43.3-55.9%), high enrichment factors (433-559), and low limits of detection (0.45-0.89 µg L-1 ). CONCLUSION: The method was applied in the analysis of pesticides in various fruit juices. Chlorpyrifos was found in peach juice at a concentration of 27 ± 2 µg L-1 (n = 3) using a gas chromatography-flame ionization detector. To verify the results, the peach juice was also injected into gas chromatography-mass spectrometry after applying the proposed extraction method. © 2022 Society of Chemical Industry.

Development of an in-syringe gas-assisted density tunable solidification of floating organic droplet-based dispersive liquid phase microextraction method coupled with HPLC-MS/MS for monitoring amikacin in biological fluids.

A new sample preparation method named in-syringe gas-assisted density tunable dispersive liquid phase microextraction based on solidification of floating organic droplet has been introduced. This method was coupled with high-performance liquid chromatography-tandem mass spectrometry and used for the extraction and quantification of amikacin in plasma and exhaled breath condensate (EBC) samples of the patients receiving amikacin. In the proposed approach, an inert gas is bubbled into a syringe barrel containing aqueous solution of the analyte and a mixture of low density extraction solvent and volatile density modifier. Consequently, the density modifier is evaporated and the analyte is migrated into the released extractant droplets. Basic parameters affecting efficiency of the developed method were optimized. Under optimum conditions, the method limits of detection were 0.06 and 0.29 ng/mL in EBC and plasma, respectively. The extraction recoveries were 90% and 87% in EBC and plasma, respectively. Also, the obtained relative standard deviations were below 9.5% and 9.8% for EBC and plasma, respectively. Considering these results, the developed method provides a quick and efficient way to determine amikacin in patients' biological fluids and can be used widely in drug monitoring and clinical studies.

Determination of carnitine ester profile in the children with type 1 diabetes: a valuable step towards a better management.

Objective: This study is designed to investigate the levels of carnitine and acylcarnitines (ACs) in the children with diabetes type 1 compared to the healthy subjects.Methods: Forty-two type 1 diabetic children and healthy subjects were recruited in the study, respectively. In addition to FBS and Hb A1C, free carnitine and ACs in butyl-ester form in the fasting blood samples were assessed by isotope dilution mass spectrometry for all diabetics and controls using the tandem mass spectrometry system.Results: Diabetic patients had a higher level of C, C4, C6, C14, C18:2, and C18:2OH. Females had elevated C14:2 compared to the males. The C18:2 and C18:2OH levels were elevated as the Hb A1C level increased. The C18:2, C14OH were mostly increased in the prediabetic and diabetic patients, respectively.Conclusion: Increased ACs level indicates the increased acyl-CoA intermediates for the fatty acids and amino acids oxidation.

Early Glycemic Control Assessment Based on Consensus CGM Metrics.

Continuous glucose monitoring (CGM) has revolutionized the world of diabetes and transformed the approach to diabetes care. In this context, an expert panel has reached consensus on clinical targets for CGM data interpretation based on eight CGM metrics. At least 70% of 14 consecutive CGM days (referred to as a period) are recommended to assess glycemic control based on the metrics. In clinical practice less CGM data may be available. Therefore, the primary aim of this study is to explore the ability to recover the consensus metrics utilizing less than 14 days of CGM data (intra-period). As a secondary aim, we investigate the recovery considering two consecutive periods (inter-period). The analyses are based on real-world CGM data from 484 diabetes users (4726 periods) acquired from the Cornerstones4Care® Powered by Glooko app. Using up to 14 accumulated days, the consensus metrics are calculated for each user and period, and compared to the fully 14 accumulated intra- and inter-period days. Relatively low deviations were observed for time in range (TIR) and average based metrics when using less than 14 days, however, we observed large deviations in metrics characterizing infrequent events such as time below range (TBR). Furthermore, the consensus metrics obtained in two consecutive 14 day periods have clear discrepancies (inter-period). Recovering consensus metrics using less than 14 days might still be valuable in terms of interpreting CGM data in certain clinical contexts. However, caution should be taken if treatment decisions would be made with less than 14 days of data on critical metrics such as TBR, since the metrics characterizing infrequent events deviate substantially when less data are available. Substantial deviation is also seen when comparing across two consecutive periods, which means that care should be taken not to over-generalize consensus metric based glycemic control conclusions from one period to subsequent periods.

COVID-19: A New Horizon in Congenital Heart Diseases.

Objective: Previous studies have demonstrated that both children and adult patients with a history of congenital heart disease (CHD) are at high risk for coronavirus disease 2019 (COVID-19) infection. This study investigates the status of COVID-19 infection among children undergoing surgical repair within the past 2 years. Methods: All alive patients operated on in a tertiary referral center between March 2018 and March 2020 were recruited in the present study. Detailed demographics, past medical and surgical history, and physical examination were reviewed for each patient. During the COVID-19 pandemic, data regarding the patient's status were collected by telephone survey from April 15 to April 30, 2020. Results: A total number of 210 patients are analyzed in this study. Participants' median age was 21.59 months [interquartile range (IQR) = 12-54.67], and 125 (59.5%) were female. The median interval between surgery and COVID-19 assessment was 305 days (IQR = 215-400). In addition, 67 (32%) patients used angiotensin receptor blocker (ARB)/angiotensin-converting enzyme (ACE) inhibitor (spironolactone and/or captopril). Sixteen patients (7.6%) were symptomatic and had positive chest CT results and/or RT-PCR compared to the previously reported prevalence of COVID-19 among the pediatric population (2.4% of children with <18 years of age); the prevalence of COVID-19 among the patients operated on due to CHD in the present study was significantly higher (p = 0.00012). Two patients were admitted to the intensive care unit (ICU); one patient was discharged 2 weeks later with acceptable status, and one patient died 2 days after ICU admission due to cardiac and respiratory arrest and myocarditis. The complexity of the underlying cardiac disorders was not different between patients with low risk (p = 0.522), suspicious patients (p = 0.920), and patients positive for COVID-19 (p = 0.234). The ARB/ACE inhibitor consumption was not associated with the COVID-19 infection [p = 0.527, crude odds ratio (OR) = 1.407, 95% CI = 0.489-4.052]. Conclusion: Children with a history of previous CHD surgery are more susceptible to infections, especially those infections with pulmonary involvements, as the lung involvement could cause worsening of the patient's condition by aggravating pulmonary hypertension. The results of the current study indicate that these patients are more prone to COVID-19 infection compared to the healthy children population.

Modeling and optimization of radish root extract drying as peroxidase source using spouted bed dryer.

The main advantages of the dried enzymes are the lower cost of storage and longer time of preservation for industrial applications. In this study, the spouted bed dryer was utilized for drying the garden radish (Raphanus sativus L.) root extract as a cost-effective source of the peroxidase enzyme. The response surface methodology (RSM) was used to evaluate the individual and interactive effects of main parameters (the inlet air temperature (T) and the ratio of air flow rate to the minimum spouting air flow rate (Q)) on the residual enzyme activity (REA). The maximum REA of 38.7% was obtained at T = 50 °C and Q = 1.4. To investigate the drying effect on the catalytic activity, the optimum reaction conditions (pH and temperature), as well as kinetic parameters, were investigated for the fresh and dried enzyme extracts (FEE and DEE). The obtained results showed that the optimum pH of DEE was decreased by 12.3% compared to FEE, while the optimum temperature of DEE compared to FEE increased by a factor of 85.7%. Moreover, kinetic parameters, thermal-stability, and shelf life of the enzyme were considerably improved after drying by the spouted bed. Overall, the results confirmed that a spouted bed reactor can be used as a promising method for drying heat-sensitive materials such as peroxidase enzyme.

Combining a liquid-liquid extraction with successive air assisted liquid-liquid microextraction for the analysis of phytosterols present in animal based butter and oil samples.

In this study, an elevated temperature liquid-liquid extraction combined method with successive air-assisted liquid-liquid microextraction has been proposed for the extraction of four phytosterols in cow milk butter and animal oil samples prior to gas chromatography-flame ionization detector. The method is started by combining a few grams of the melted butter or oil samples with ethanol. The mixture is vortexed and placed into a water-bath adjusted at 50 °C. After a few minutes, the mixture is allowed to cool at room temperature. In this step, the butter or oil is become stiff and ethanol is collected on top of the sample. The separated ethanol phase is collected and mixed with deionized water to obtain a homogenous solution. After that, a few microliters of ethyl methyl ammonium chloride: pivalic acid deep eutectic solvent is added into the solution and the mixture was pulled into a glass test tube and pushed back to the tube for five times. After centrifugation, whole of the collected phase at the bottom of tube was withdrawn and transferred into a microtube and contacted with sodium hydroxide solution. The mixture is withdrawn and released to the tube 2 times to remove the extracted fatty acids. The validation data verified that high enrichment factors (385-450) and extraction recoveries (77-90%), low limits of quantification (2.6-5.2 ng g-1) and detection (0.73-1.5 ng g-1), and satisfactory relative standard deviations (≤ 9.3%) can be obtained with this method. At last, the developed method was successfully used for the analysis of phytosterols in various butter and oil samples marketed in Tabriz, Iran.