Tetraselmis chuii Edible Microalga as a New Source of Neuroprotective Compounds Obtained Using Fast Biosolvent Extraction.

Tetraselmis chuii is an EFSA-approved novel food and dietary supplement with increasing use in nutraceutical production worldwide. This study investigated the neuroprotective potential of bioactive compounds extracted from T. chuii using green biobased solvents (ethyl acetate, AcOEt, and cyclopentyl methyl ether, CPME) under pressurized liquid extraction (PLE) conditions and supercritical fluid extraction (SFE). Response surface optimization was used to study the effect of temperature and solvent composition on the neuroprotective properties of the PLE extracts, including anticholinergic activity, reactive oxygen/nitrogen species (ROS/RNS) scavenging capacity, and anti-inflammatory activity. Optimized extraction conditions of 40 °C and 34.9% AcOEt in CPME resulted in extracts with high anticholinergic and ROS/RNS scavenging capacity, while operation at 180 °C and 54.1% AcOEt in CPME yielded extracts with potent anti-inflammatory properties using only 20 min. Chemical characterization revealed the presence of carotenoids (neoxanthin, violaxanthin, zeaxanthin, α- and ß-carotene) known for their anti-cholinesterase, antioxidant, and anti-inflammatory potential. The extracts also exhibited high levels of omega-3 polyunsaturated fatty acids (PUFAs) with a favorable ω-3/ω-6 ratio (>7), contributing to their neuroprotective and anti-inflammatory effects. Furthermore, the extracts were found to be safe to use, as cytotoxicity assays showed no observed toxicity in HK-2 and THP-1 cell lines at or below a concentration of 40 µg mL-1. These results highlight the neuroprotective potential of Tetraselmis chuii extracts, making them valuable in the field of nutraceutical production and emphasize the interest of studying new green solvents as alternatives to conventional toxic solvents.

Separation and identification of different therapeutic pharmaceutical drugs on humic acid-based HPLC column: Method optimization using central composite design.

Multimode chromatographic separations are highly desirable in pharmaceutical and environmental sciences. Current study deals with the application of newly developed mixed-mode end capped-immobilized humic acid onto an aminopropyl silica based chromatographic column for separation and identification of six drugs belonging to different therapeutic groups for its applicability in pharmaceutical industries. For this, central composite design was used to evaluate the separation and resolution by optimization of three most effective parameters (acetonitrile%, flow rate, and pH of mobile phase). Second-order quadratic model was used to evaluate their effect on resolution of peaks; the probability value (<0.05) obtained from analysis of variance suggested the best applicability of the model. Desirability function was applied to calculate optimum conditions (44.8% acetonitrile, 1.75 mL/min of flow rate, and 7.5 pH) required to achieve maximum separation with good resolution within 11 min. The method was validated for linearity, precision accuracy, selectivity, and sensitivity. The results revealed a highly precise (coefficient of variance > 1%), linear (R2  = 0.99), and highly selective method. Moreover, the limit of detection/quantification values revealed acceptable sensitivity of the method. The developed column was compared for its efficiency with a commercially available column and found to be highly applicable for industrial applications.

Frequency, risk factors, and impacts on quality of life of the restless legs syndrome and side effects among antidepressant users in a tertiary hospital: an observational cross-sectional study.

Restless leg syndrome (RLS) is a common but underestimated sensorimotor disorder that significantly affects the quality of life (QoL) which can be induced by antidepressants. This study aims to investigate the frequency and potential risk factors of RLS and side effects in selective serotonin reuptake inhibitors/serotonin and noradrenaline reuptake inhibitors (SSRI/SNRI) users. This cross-sectional study included 198 outpatients who received SSRI/SNRI for 4-8 weeks. Clinical evaluation was performed using the International Restless Leg Syndrome Study Group rating scale for RLS, Udvalg for Kliniske Undersøgelser side effects rating scale, and a short form 36 (SF-36) questionnaire for QoL. The frequency of RLS was 25%. RLS significantly increased with smoking and habituality. Also, habituality increased neurologic side effects reporting. The use of antipsychotics and calcium channel blockers decreased reporting of autonomic side effects. QoL decreased with RLS, psychiatric, neurologic, autonomic, and other side effects in different domains of SF-36. These findings suggested that SSRI/SNRI use could be associated with a higher risk of RLS, especially in smokers. QoL could be influenced negatively by RLS and all side effects. However, further prospective studies are needed to confirm these associations in large samples.

The importance of length and area estimate for forensic medicine without using a measurement tool.

Background/aim: Accurately measuring and recording the length or area of lesions affects the judgement of the forensic report, surgical wound management and, in some countries, the billing of health care services. The aim of this study was to determine whether the length and area of lesions described by physicians by estimation are accurate. Materials and methods: This study was designed as a cross-sectional descriptive study and was conducted with 494 participants consisting of internists and physicians at Ondokuz Mayis University, Faculty of Medicine. The participants were asked to estimate the lengths or areas of 6 different shapes in the questionnaire form without using a measuring instrument. Results: Of the participants, 216 (43.7%) were interned physicians and 278 (56.3%) were physicians. Most 122 people (24.7%) answered the curved line shape with a length of 4 cm as "exact value". The average of the values given by the internists to each shape was higher than the average of the values given by the physicians to each shape and was found to be further away from the true value. It was determined that more than half of the participants gave values above the actual length and area values of the shapes. It was observed that the rate of correct estimation of line shapes was higher than the rate of correct estimation of area shapes both as exact values and with ±10% margin of error. For line shapes, it was observed that the number of those who gave exact values decreased as the line length increased. Conclusion: When preparing forensic reports, determining surgical wound management and billing, estimated data should not be used in lesion description. It would also be useful to develop tools that will enable physicians to make measurements in terms of easy use.

Chromatographic evaluation of tocols and sterols of processed canola oil and deodorizer distillate.

Tocopherols and tocotrienols in the combined form are known as tocols. Changes of total and individual tocols and sterols concentration of canola oil and deodorizer distillate (DD) during different processing stages were evaluated with the application of gas chromatography (GC) and high-performance liquid chromatography (HPLC). For sterols analysis, GC coupled with flame ionization detector (FID) was used while tocols in canola oil samples and DD, normal phase (NP) HPLC was applied. The results of the present study indicated that levels of total and individual tocols and sterols content were decreased during processing (neutralization to deodorization). Deodorization was found to be the most effective process for the reduction of total sterols and tocols as 55.9% and 34.2%, respectively. A high amount of tocols and sterols was observed in DD. Among tocols and sterols; beta tocopherol (ß-T) and ß-sitosterol were found to be in greater concentration 53.97% and 31.82%, respectively. Therefore, DD could be used as a valuable by-product in the cosmetics and food industries.

The Relationship of Serum Leptin and Ghrelin Levels with Craving and Withdrawal in Opioid Use Disorder.

Objective: This study aims to investigate plasma levels of leptin, acyl ghrelin, and unacylated ghrelin during heroin withdrawal in patients with opioid use disorder (OUD) with regard to the relationship of these levels with craving and their changes over time. Methods: This study included 28 male patients diagnosed with OUD according to DSM-5 diagnostic criteria who received inpatient rehabilitation. The control group included 28 healthy male volunteers with characteristics similar to the patient group. Plasma leptin, acyl ghrelin, and unacylated ghrelin levels of the patients were measured 3 times throughout the study by collecting blood on the first day, the seventh day at the end of the detox, and the twenty-first day. Blood was collected only once from the control group to determine their plasma leptin, acyl ghrelin, and unacylated ghrelin levels. Results: Our study did not determine any statistically significant differences between patients with OUD and healthy controls with regard to plasma leptin, acyl ghrelin, and unacylated ghrelin levels on the first, seventh, and twenty-first days of withdrawal. Plasma levels of leptin, acyl ghrelin, and unacylated ghrelin did not significantly correlate with craving scores. Conclusion: This study does not support the hypothesis that plasma leptin, acyl ghrelin, and unacylated ghrelin levels are markers in those with OUD. Further research, particularly in humans, is recommended to replicate and expand on the findings of the current literature.

Processing impact on tocopherols and triglycerides composition of soybean oil and its deodorizer distillate evaluated by high-performance liquid chromatography.

In the present study, high-performance liquid chromatography (HPLC) was used for the separation of tocopherols and triglycerides of processed soybean oil and deodorizer distillate (DD). The results of normal and reversed-phase modes of HPLC revealed that concentrations of tocopherols and triglycerides content were decreased during the neutralization, bleaching, and deodorization processes. The loss of individual tocopherols ranged between 55.16% and 63.25%. During processing, triglycerides containing stearic-oleic-linoleic (SOL) moieties and palmitic-palmitic-linoleic (PPL) fragments showed greater reduction up to 38.14% and 37.69%, respectively. Among tocopherols and triglycerides; γ-tocopherol and oleic-oleic-oleic (OOO) were found to be in greater concentrations 5.53% and 19.78%, respectively in DD as compared to their counterparts. A maximum reduction of tocopherols was observed in the deodorization step. DD was found to be a rich source of bioactive components; therefore, it could be used for many industrial applications including pharmaceutical formulations, cosmetics, and food industries.

Intravenous buprenorphine/naloxone and concomitant oral pregabalin misuse: a case report.

Opioid misuse and dependence are major medical and social concerns worldwide. Buprenorphine/naloxone combination (BNC) is a drug that has misuse potential and is used to treat opioid dependence, including buprenorphine and naloxone. Buprenorphine shows its pharmacological effects by binding to opioid receptors. Buprenorphine is a partial agonist and has smaller maximal effects compared to those of full agonists (heroin, methadone). Naloxone is a non-selective opiate antagonist added to buprenorphine for the prevention of intravenous diversion. BNC is used in the treatment of opioid dependence for detoxification and maintenance. The drug should be used as a sublingual film tablet. Pregabalin is used in the treatment of neuropathic pain, epilepsy and anxiety disorders. It is increasingly being reported as possessing a potential for misuse. In this article, we present a case of intravenous BNC and concomitant oral pregabalin misuse that developed in a monitored and treated patient for the reason of opioid dependence.

New potential humic acid stationary phase toward drug components: Development of a chemometric-assisted RP-HPLC method for the determination of paracetamol and caffeine in tablet formulations.

A new humic acid based stationary phase has been used, for the first time, to achieve the separation and quantification of paracetamol and caffeine in pharmaceutical preparations under reversed-phase high-performance liquid chromatography conditions. Central composite design was applied as a powerful tool to optimize the most dominant parameters that influence the resolution of reversed-phase high-performance liquid chromatography, that is, mobile phase composition (acetonitrile percentage in water), flow rate, and column temperature. The optimum conditions were obtained as 21.69%, 1.5 mL/min, and 15°C, respectively, with the aid of a second-order quadratic model and desirability function. Under the optimum conditions, the peaks could be baseline separated within 10 min. For the developed reversed-phase high-performance liquid chromatography method, the linearity was investigated in the concentration ranges of 2-160 mg/mL (R(2) = 0.999) for paracetamol and 2-9.9 mg/mL (R(2) = 0.991) for caffeine. Mean recoveries for paracetamol and caffeine were 95.90 and 95.68%, respectively. The limits of detection and quantification were 4.1 × 10(-4) and 1.3 × 10(-3) mg/mL for paracetamol and 1.6 × 10(-4) and 5.0 × 10(-4) mg/mL for caffeine. The results showed that the new humic acid based stationary phase is very suitable for the separation of paracetamol and caffeine in pharmaceutical preparations and, thus it can be used effectively in the pharmaceutical industry.

A simplified FTIR chemometric method for simultaneous determination of four oxidation parameters of frying canola oil.

Transmission Fourier transform infrared (FTIR) spectroscopic method using 100 µm KCl cell was applied for the determination of total polar compounds (TPC), carbonyl value (CV), conjugated diene (CD) and conjugated triene (CT) in canola oil (CLO) during potato chips frying at 180 °C. The calibration models were developed for TPC, CV, CD and CT using partial least square (PLS) chemometric technique. Excellent regression coefficients (R(2)) and root mean square error of prediction values for TPC, CV, CD and CT were found to be 0.999, 0.992, 0.998 and 0.999 and 0.809, 0.690, 1.26 and 0.735, respectively. The developed calibration models were applied on samples of canola oil drawn during potato chips frying process. A linear relationship was obtained between CD and TPC with a good correlation of coefficient (R(2)=0.9816). Results of the study clearly indicated that transmission FTIR-PLS method could be used for quick and precise evaluation of oxidative changes during the frying process without using any organic solvent.

Occurrence of ochratoxin A in poultry feeds and feed ingredients from Pakistan.

A study was conducted to evaluate the occurrence of ochratoxin A (OTA) in complete poultry feeds (n=80) and poultry feed ingredients (n=286) from Pakistan. All samples were first analyzed by indirect enzyme linked immunosorbent assay (ELISA), samples with OTA concentrations above the European Union maximum regulatory limit (MRL, 100 µg/kg) were further confirmed by HPLC-FLD. Contamination frequency and mean OTA levels were 31% and 51 µg/kg in feed ingredients, and the corresponding values for complete feeds were 38% and 75 µg/kg. Ten samples of complete poultry feed and 19 samples of feed ingredients contained OTA at levels higher than the MRL. The results of the present study indicate that there is a strong need for a more intense monitoring programs for OTA in poultry feed.

Application of multivariate chemometric techniques for simultaneous determination of five parameters of cottonseed oil by single bounce attenuated total reflectance Fourier transform infrared spectroscopy.

Single bounce attenuated total reflectance (SB-ATR) Fourier transform infrared (FTIR) spectroscopy in conjunction with chemometrics was used for accurate determination of free fatty acid (FFA), peroxide value (PV), iodine value (IV), conjugated diene (CD) and conjugated triene (CT) of cottonseed oil (CSO) during potato chips frying. Partial least square (PLS), stepwise multiple linear regression (SMLR), principal component regression (PCR) and simple Beer׳s law (SBL) were applied to develop the calibrations for simultaneous evaluation of five stated parameters of cottonseed oil (CSO) during frying of French frozen potato chips at 170°C. Good regression coefficients (R(2)) were achieved for FFA, PV, IV, CD and CT with value of >0.992 by PLS, SMLR, PCR, and SBL. Root mean square error of prediction (RMSEP) was found to be less than 1.95% for all determinations. Result of the study indicated that SB-ATR FTIR in combination with multivariate chemometrics could be used for accurate and simultaneous determination of different parameters during the frying process without using any toxic organic solvent.

The performance of BALISTIKA 2010 system for 9 mm × 19 mm and 7.65 mm × 17 mm cartridge case correlation.

The firearm identification has two examination phases; the first phase is "one by one" cartridge case or bullet identification. The second phase is "Open Case File (OCF)" examination. Due to the size of the OCF archive, the OCF examination with only comparison microscopes takes a long time and is an unfeasible process. The Computerized Ballistic Identification Systems (CBIS) has become an essential tool for archive examination by correlation and preliminary eliminations. In this study, two objectives were pursued; the first is measuring the performance of the BALISTIKA 2010 system on cartridge case acquisition of handguns, correlation and examination. The second objective is the examination of the performance on the correlation according to brand and models of firearms. Detailed experimental results are demonstrated for about 2000 cartridge cases.

Rapid determination of free fatty acid content in waste deodorizer distillates using single bounce-attenuated total reflectance-FTIR spectroscopy.

A simple, rapid, economical, and environmentally friendly analytical method was developed for the quantitative assessment of free fatty acids (FFAs) present in deodorizer distillates and crude oils by single bounce-attenuated total reflectance-FTIR spectroscopy. Partial least squares was applied for the calibration model based on the peak region of the carbonyl group (C=O) from 1726 to 1664 cm(-1) associated with the FFAs. The proposed method totally avoided the use of organic solvents or costly standards and could be applied easily in the oil processing industry. The accuracy of the method was checked by comparison to a conventional standard American Oil Chemists' Society (AOCS) titrimetric procedure, which provided good correlation (R = 0.99980), with an SD of +/- 0.05%. Therefore, the proposed method could be used as an alternate to the AOCS titrimetric method for the quantitative determination of FFAs especially in deodorizer distillates.

Towards multimodal HPLC separations on humic acid-bonded aminopropyl silica: RPLC and LEC behavior.

In the present study, metal binding property of humic acid (HA) was successfully adapted to the ligand-exchange concept, and metal-loaded immobilized humic acid was used as a ligand exchanger stationary phase for separation of some nucleosides. Humic-acid bonded aminopropyl silica (EC-HA-APS) was turned into ligand exchanger forms by loading aqueous solutions of Cu(2+), and Co(2+) to the column (4.6 × 150; as mm) packed with EC-HA-APS. Metal ion solutions were loaded to the column in a stepwise manner where the concentration of metal ion solution being loaded to the column was increased gradually between 5 and 100mM. The progress of metal loading process was monitored via the breakthrough curves propagated stepwise. Ligand-exchange chromatography (LEC) studies were performed on an HPLC system, and chromatographic behaviors of the studied nucleosides (i.e. uridine, Urd; thymidine, Tyd; cytidine, Cyd; adenosine, Ado; and guanosine, Guo) were investigated on Cu(2+) and Co(2+) loaded forms of the EC-HA-APS (Cu-EC-HA-APS and Co-EC-HA-APS). Effect of mobile phase composition, temperature, and the type of metal ion loaded to the column on the retentive behaviors of the compounds was studied, in detail. The studied solutes exhibited mixed-mode RPLC/LEC behavior on the stationary phase. Metal-loaded column (M-EC-HA-APS) was easily regenerated into its original form, EC-HA-APS, with 98 ± 2% metal recoveries, by using aqueous mixture of EDTA+NH(3) at pH=7.5. Thus, the stationary phase exhibited a high flexibility between RPLC and LEC modes. This property, also, made it possible to convert the stationary phase into various ligand exchanger forms by loading different metal ions. Hence, capacity and selectivity of the stationary phase towards the studied species was manipulated easily by loading different metal ions to the stationary phase. Baseline separation for the studied species was achieved on Cu-EC-HA-APS and Co-EC-HA-APS and some differentiations were observed in capacity and selectivity, depending on the type of metal loaded. Thus, being as the first endeavor on usability of immobilized HA as a ligand exchanger stationary phase, the present study is believed to be useful to understand multifunctional character of HA-based solid/stationary phases.

A novel approach for determination of free fatty acids in vegetable oils by a flow injection system with manual injection.

A non-aqueous flow injection method for determining free fatty acid (FFA) content in corn and sunflower oil samples was developed. A single-line manifold system was built by modification of an HPLC for flow injection analysis (FIA). Without pre-treatment, oil samples were injected into a n-propanol solution containing KOH and phenolphthalein (PHP). The main parameters, such as flow rate of carrier phase, length, geometry, inner diameters of the coils and reagent concentration were all optimized. The proposed FIA method was validated for precision, accuracy, linear region, limit of detection (LOD) and limit of quantification (LOQ). The intra- and inter-day measurements of the precision of the method were found to be within the limits of acceptance criteria (RSD < 1%), and were rugged when the method was performed by a different analyst. The linear concentration range was calculated as 0.09-1.50 and 0.07-1.40 FFA% for corn and sunflower oils, correspondingly. The LOD and LOQ were found to be 7.53 × 10(-4)-2.28 × 10(-3) oleic acid % and 7.11 × 10(-4)-2.23 × 10(-3) oleic acid % for corn and sunflower oils, respectively. The results were compared with those obtained by the AOCS (Ca-5a-40) method using statistical t and F tests, and a significant difference was not observed between the methods at a 95% confidence level. The proposed method is suitable for quality control of routine applications due to its simplicity, high sample throughput, and economy of solvents and sample, offering considerable promise as a low cost analytical system that needs minimum human intervention over long periods of time.

Towards multimodal HPLC separations on humic acid-bonded aminopropyl silica: RPLC and HILIC behavior.

The stationary phase characteristics of the material obtained through immobilization of humic acid (HA) to aminopropyl silica (APS) via amide-bond formation were investigated. The material was characterized in terms of elemental analysis, FTIR, thermogravimetric analyses, pH point of zero charge measurements, potentiometric titrations, and contact angle measurements. Amount of HA bonded to APS was determined from the elemental analysis results, and found as 170 mgHA/gAPS. Stability of the material was studied in aqueous media at different pH values, and amount of HA released at pH=8 did not exceed 2% of the total immobilized HA. Stationary phase characteristics of the well-characterized material were investigated in an HPLC system by using some low-molecular weight polar compounds (i.e. some nucleosides and nucleobases) as test solutes. Effect of some experimental variables such as column conditioning, composition of mobile phase, and temperature on the chromatographic behavior of the studied compounds was studied. Role of ammonium solutions at different pH values on retentive properties of the species was also studied. Retention factors (k') versus volume percentage of organic modifier exhibited a U-curve, which was evaluated as an indication for RPLC/HILIC mixed-mode behavior of the stationary phase. Orthogonality between RPLC and HILIC modes was analyzed through geometric approach, and found as 48.5%. Base-line separation for the studied groups of compounds was achieved under each studied mode, and some differentiations were observed in elution order of the compounds depending on the HPLC mode applied. Chromatograms recorded under RPLC and HILIC modes were compared with those recorded on APS under similar conditions, and thus the influence/importance of HA immobilization process was evaluated in detail. In light of the obtained results, immobilized HA is represented as a useful stationary phase for HPLC separations.

Determination of defensin HNP-1 in human saliva of patients with oral mucosal diseases.

Saliva is a biological fluid that is easily obtainable and that can give useful information both in systemic and oral diseases. In this study, a chromatographic method was applied to determine the amount of defensin HNP-1 in human saliva of patients with oral mucosal diseases before and after treatments and compared with controls. Defensin human neutrophil peptide-1 (HNP-1) was identified and confirmed. The concentration of HNP-1 in saliva was determined by comparing the area of eluted HNP-1 with that of HNP-1 standard. Linear calibration range of defensin HNP-1 was 0.10 to 0.90 µg/10 µL with R(2) values of 0.996. The concentrations of HNP-1 in the saliva of patients with oral lichen planus, Behçet's disease, and recurrent apthous stomatitis were 33.6 ± 10.6, 15.5 ± 7.6, and 36.3 ± 9.5 µg mL(-1) (mean ± S.D.), respectively. The salivary defensin-1 concentration was significantly higher in patients with oral mucosal diseases than in healthy volunteers; furthermore, in patients with oral mucosal diseases, the concentration was significantly higher before treatment than after treatment.

Monitoring of some organochlorine pesticide residues of butter in Konya, Turkey.

Some organochlorine pesticide residues were investigated in the samples of all local commercial butter brands sold in the supermarket in Konya (Turkey). Some of the samples were found to have the DDT complex (DDT, DDD, DDE and isomers), total HCH complex (alpha-HCH, beta-HCH, gamma-HCH), aldrin, dieldrin and endosulfan (I and II). Nearly 94% of the butter samples were found to be contaminated. Endrin and heptachlorine were detected in three and two samples, respectively, while 87% of samples were contaminated by one or more HCH isomers. 78% of the organochlorine pesticide was DDT and/or its metabolites. These results confirmed that butters sold in Konya presented organochlorine pesticide residues indicating a human exposure.