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.

Lead poisoning as a differential diagnosis in pediatric patients with chronic abdominal pain: a case-control study in Tehran-Iran.

BACKGROUND: Chronic abdominal pain is a potential symptom of lead poisoning, which is often challenging to diagnose. This case-control study aimed to evaluate blood lead levels in pediatric patients with chronic abdominal pain. METHODS: The case-control study was conducted on 190 pediatrics who presented to the Children's Medical Center Hospital clinics, Tehran between April 2021- 2023. The children were divided into two groups: the case group, consisting of 81 patients with chronic abdominal pain, and the matched control group; 109 children without any gastrointestinal symptoms. The statistical analysis of the data was performed using STATA 16. A multiple logistic regression model was used to assess the association of different independent variables with chronic abdominal pain. RESULTS: There was no significant difference between mean (± standard deviation [SD]) of age (8.80(2.7) years vs. control group: 9.23(3.9) years), sex, and BMI (16.55(4.6) vs. 17.32(4.7)) of the patients with chronic abdominal pain (case group) and the control group, whereas the mean weight was remarkably low in patients with chronic abdominal pain: 27.25(± 12.1) kg vs. 31.70(± 14.7) kg (P value = 0.028). Fifty-nine percent of children with chronic abdominal pain had serum lead levels ≥ 10 µg/dL. The mean (SD) of blood lead levels was statistically high in the case group: 11.09 (± 5.35) µg/dL vs. control group: 8.26 (± 5.01) µg/dL) (P value ≤ 0.05). The appetite level was significantly low in the case group: 3.8 (± 2.5) vs. control group 5.4 (± 1.3). CONCLUSIONS: Lead poisoning could be a possible cause of children's chronic abdominal pain. Regarding the high rate of lead poisoning in children exerting appropriate measures to reduce their exposure to lead is necessary.

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.

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 ).

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.

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.

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.

Combination of temperature-assisted ternary phase homogenous liquid-liquid extraction with deep eutectic solvent-based dispersive liquid-liquid microextraction for the extraction of phytosterols from cow milk and cream samples.

A three-phase homogenous liquid-liquid extraction method followed by deep eutectic solvent-based dispersive liquid-liquid microextraction has been used for the extraction and preconcentration of some phytosterols from cow milk and cream samples. In this method, cow milk or melted cream was transferred into a glass test tube and a mixture of sodium hydroxide solution (as fatty acids saponification agent) and acetonitrile (as extraction/dispersive solvent) was added to the solution. The mixture was shaken manually and placed in a water bath. After dissolving sodium chloride and centrifugation, the obtained supernatant phase (acetonitrile) was removed and mixed with ethyl methyl ammonium chloride: phenyl acetic acid deep eutectic solvent at microliter level. Under optimum conditions, low limits of detection (1.6-4.1 µg/L) and quantification (5.3-13.1 µg/L), high enrichment factors (138-207), and extraction recoveries (55-83%), and good precision (relative standard deviations ≤ 6.8%) can be obtained. The method was done on different milk and cream samples and the results showed that total phytosterols level in all samples were in the permitted level established by the National Standard Organization Guidelines, except for three cream samples.

Development of a liquid-nitrogen-induced homogeneous liquid-liquid microextraction of Co(II) and Ni(II) from water and fruit juice samples followed by atomic absorption spectrometry detection.

In this study, a simple and rapid sample preparation method named liquid-nitrogen-induced homogeneous liquid-liquid microextraction has been developed for the extraction and pre-concentration of Co(II) and Ni(II) ions before their analysis by flame atomic absorption spectrometry. For this purpose, first, acetonitrile containing 8-hydroxyquinoline is added into a sample solution and the mixture is vortexed. As a result, a homogeneous solution is formed. Subsequently, the solution is cooled using liquid nitrogen for a few seconds. By this process, due to difference in the freezing point of acetonitrile and water, the homogeneous state is broken and the analytes (as oxinate complexes) are extracted into liquid acetonitrile phase collected on top of the frozen aqueous phase. The linear dynamic ranges obtained for Ni(II) and Co(II) were 1.0-30 and 0.50-20 µg L-1, respectively. The obtained limits of detection were 0.36 and 0.20 µg L-1 for Ni(II) and Co(II), respectively.

A three-phase solvent extraction system combined with deep eutectic solvent-based dispersive liquid-liquid microextraction for extraction of some organochlorine pesticides in cocoa samples prior to gas chromatography with electron capture detection.

A sample pretreatment method based on the combination of a three-phase solvent extraction system and deep eutectic solvent-based dispersive liquid-liquid microextraction has been introduced for the extraction of four organochlorine pesticides in cocoa samples before their determination by gas chromatography-electron capture detection. A mixture of sodium chloride, acetonitrile, and potassium hydroxide solution is added to cocoa bean or powder. After vortexing and centrifugation of the mixture, the collected upper phase (acetonitrile) is removed and mixed with a few microliters of N,N-diethanol ammonium chloride: pivalic acid deep eutectic solvent. Then it is rapidly injected into deionized water and a cloudy solution is obtained. Under optimum conditions, the limits of detection and quantification were found to be 0.011-0.031 and 0.036-0.104 ng/g, respectively. The obtained extraction recoveries varied between 74 and 92%. Also, intra- (n = 6) and interday (n = 4) precisions were less than or equal to 7.1% for the studied pesticides at a concentration of 0.3 ng/g of each analyte. The suggested method was applied to determine the studied organochlorine pesticide residues in various cocoa powders and beans gathered from groceries in Tabriz city (Iran) and aldrin and dichlobenil were found in some of them.

Development of organic solvents-free mode of solidification of floating organic droplet-based dispersive liquid-liquid microextraction for the extraction of polycyclic aromatic hydrocarbons from honey samples before their determination by gas chromatography-mass spectrometry.

In this study, a green mode of solidification of floating organic droplet-based dispersive liquid-liquid microextraction has been developed for the extraction of 16 polycyclic aromatic hydrocarbons from honey samples before their determination by gas chromatography-mass spectrometry. In this method, an appropriate volume of menthol:decanoic acid deep eutectic solvent (as an extraction solvent) is added on a sugar cube (as a disperser agent). In the following, the cube is released into the diluted honey sample placed in a tube. After manual shaking a cloudy state is obtained as a result of dispersing the extraction solvent droplets throughout the sample solution and the analytes are extracted into them. After placing the tube in an ice bath, the droplet of the extractant is solidified on the top of the solution. This drop is taken and after dissolving in acetonitrile, an aliquot of the solution is injected into the separation system. Under optimum conditions, the suggested approach had high extraction recoveries (76-93%) and enrichment factors (380-465), low limits of detection (14-52 ng/kg) and quantification (47-173 ng/kg), and satisfactory repeatability (relative standard deviation ≤ 9%).

Development of a stir bar sorptive extraction method coupled to solidification of floating droplets dispersive liquid-liquid microextraction based on deep eutectic solvents for the extraction of acidic pesticides from tomato samples.

A stir bar sorptive extraction method coupled with deep eutectic solvent based solidification of floating organic droplets-dispersive liquid-liquid microextraction has been used for the simultaneous derivatization and extraction of some acidic pesticides in tomato samples. In this method, initially the analytes are adsorbed on a coated stir bar from tomato juice filled in a narrow tube. After extraction, the stir bar is removed and a water-miscible deep eutectic solvent is used to elute the analytes. Afterward, a derivatization agent and a water-immiscible deep eutectic solvent (as an extraction solvent) with melting point near to room temperature are added to the obtained eluant at µL-levels and the obtained mixture is rapidly injected into deionized water. Under the optimum conditions, the introduced method indicated high enhancement (1543-3353) and enrichment (2530-2999) factors, low limits of detection (7-14 ng/L) and quantification (23-47 ng/L), good linearity (r2 ≥ 0.9982), and satisfactory repeatabilities (relative standard deviation ≤12% for intra- and inter-day precisions at a concentration of 100 ng/L of each analyte). Finally, the proposed method was applied in analysis of the analytes in tomato samples.

Combination of a modified quick, easy, cheap, efficient, rugged, and safe extraction method with a deep eutectic solvent based microwave-assisted dispersive liquid-liquid microextraction: Application in extraction and preconcentration of multiclass pesticide residues in tomato samples.

In this study, a new two-step extraction procedure based on the combination of a modified quick, easy, cheap, effective, rugged, and safe extraction method with a deep eutectic solvent based microwave-assisted dispersive liquid-liquid microextraction has been developed for the extraction of multiclass pesticides in tomato samples before their analysis by gas chromatography with flame ionization detection. In this method, initially, an aliquot of tomato is crushed and diluted with deionized water. The mixture is then passed through a filter paper and its residue and aqueous phase are separated. Afterwards, acetonitrile as an extraction/disperser solvent is passed through the filter paper containing the refuse. The analytes remained in the refuse are extracted into the acetonitrile and then the obtained extract is mixed with a deep eutectic solvent. The obtained mixture is injected into the tomato juice and placed in a microwave oven for 15 s. Consequently, a cloudy state is formed and the extractant containing the analytes are sedimented at the bottom of the tube after centrifugation. Finally, 1 µL of the sedimented phase is removed and injected into the separation system. Under the optimum conditions, limits of detection and quantification were in the ranges of 0.42-0.74 and 1.4-2.5 ng/g, respectively.

Development of green sodium sulfate-induced solidification of floating organic droplets-dispersive liquid phase microextraction method: Application to extraction of four antidepressants.

Sodium sulfate-induced deep eutectic solvent-based solidification of floating organic droplets-dispersive liquid phase microextraction was developed prior to gas chromatography-mass spectrometry. In this method, a mixture of Na2 SO4 solution (as phase separation agent and disperser) containing menthol-decanoic acid was rapidly injected into an alkaline aqueous solution containing Na2 SO4 . The solution was placed in an ice bath and the menthol-decanoic acid solvent was solidified on the surface of the solution. Under the optimal conditions, the enrichment factors and extraction recoveries were 122-147 and 74-89%, respectively. Finally, an aliquot of the collected organic phase was removed and mixed with acetonitrile and injected into the separation system. The limits of detection and lower limits of quantification were obtained at the ranges of 13-25 and 24-41 ng L-1 , respectively. The relative standard deviations of the proposed method were ≤11% for intra- and inter-day precisions at four concentrations.

Application of a clean-up procedure using a ternary liquid phase system combined with pre-concentration by microextraction in the analysis of seven pesticides from soya milk.

BACKGROUND: A method has been developed based on a three-phase system, followed by dispersive liquid-liquid microextraction for the extraction of seven pesticides from soya milk prior to analysis by gas chromatography-flame ionization detection. The base of this method is the different extraction capability of the components of soya milk according to each of the phases involved. In this procedure, a homogeneous solution consisting of soya milk and a water-miscible solvent (acetonitrile) is separated into two phases in the presence of Na2 SO4 and the analytes are extracted into the produced acetonitrile droplets. The acetonitrile phase is mixed with a pre-concentration solvent to perform the next microextraction procedure for further enrichment of the analytes. RESULTS: Limits of detection and quantification were reached in the ranges of 0.11-0.35 and 0.35-1.20 µg L-1 , respectively. Enrichment factors and extraction recoveries were in the ranges of 562-933 and 56-93%, respectively. Relative standard deviations were ≤7% for intra- (n = 6) and inter-day (n = 5) precisions at two concentrations of 10 and 50 µg L-1 of each analyte. CONCLUSION: The proposed method was applied to the analysis of pesticides in soya milk samples at µg L-1 concentrations. © 2019 Society of Chemical Industry.

Predicting coronary artery occlusion risk from noninvasive images by combining CFD-FSI, cGAN and CNN.

Wall Shear Stress (WSS) is one of the most important parameters used in cardiovascular fluid mechanics, and it provides a lot of information like the risk level caused by any vascular occlusion. Since WSS cannot be measured directly and other available relevant methods have issues like low resolution, uncertainty and high cost, this study proposes a novel method by combining computational fluid dynamics (CFD), fluid-structure interaction (FSI), conditional generative adversarial network (cGAN) and convolutional neural network (CNN) to predict coronary artery occlusion risk using only noninvasive images accurately and rapidly. First, a cGAN model called WSSGAN was developed to predict the WSS contours on the vessel wall by training and testing the model based on the calculated WSS contours using coupling CFD-FSI simulations. Then, an 11-layer CNN was used to classify the WSS contours into three grades of occlusions, i.e. low risk, medium risk and high risk. To verify the proposed method for predicting the coronary artery occlusion risk in a real case, the patient's Magnetic Resonance Imaging (MRI) images were converted into a 3D geometry for use in the WASSGAN model. Then, the predicted WSS contours by the WSSGAN were entered into the CNN model to classify the occlusion grade.

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.

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.

Quantification and Correlation Analysis of Bacteroides Species with Diabetes-Related Amino Acids in Individuals with Prediabetes and Type 2 Diabetes Mellitus.

Background: The relationship between gut microbiota and diabetes-related amino acids significantly impacts insulin resistance and obesity. We aimed to quantify two Bacteroidetes species and their correlation with branched-chain amino acids, aromatic amino acids, and glutamate in prediabetes (preDM) and type 2 diabetes mellitus (T2DM). Methods: Fecal samples were collected from 68 participants, including 21 with T2DM, 23 with preDM, and 24 with normal glycemic tolerance (NGT). The abundance of Bacteroides vulgatus and Bacteroides thetaiotaomicron was determined by quantitative real-time polymerase chain reaction. Plasma amino acid measurements were performed using liquid chromatography coupled with tandem mass spectrometry. Results: The quantities of B. vulgatus and B. thetaiotaomicron were reduced in preDM and T2DM than in NGT subjects, but it was not statistically significant. The concentrations of leucine, valine, and tyrosine were significantly higher in preDM and T2DM than in NGT subjects (P < 0.05). A negative correlation was observed between B. thetaiotaomicron abundance and two aromatic amino acids (tyrosine, r = -0.28, P = 0.04; phenylalanine, r = -0.26, P = 0.05). Conclusions: These findings imply that, since gut microbiota varies throughout ethnic groups, further research with many participants will be required to determine the abundance of B. vulgatus and B. thetaiotaomicron in preDM and T2DM and their association with diabetes-related amino acids.

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.