Patients Survival After Paraquat Poisoning: A Report of Three Cases

Background: Paraquat poisoning is a common and often fatal herbicide poisoning in society. This study presents a clinical case series of patients who survived after paraquat poisoning.Cases Presentation: This study evaluated patients hospitalized between March 2016 and March 2021 with paraquat (PQ) poisoning who survived. Out of 115 patients with PQ poisoning, three cases of severe toxicity with an average age of 24.33 years are presented here. The urinary sodium dithionate test result was positive in all three surviving patients. All patients arrived at the poisoning emergency center within an hour of ingestion and received gastric lavage and charcoal therapy. They were also treated with corticosteroids, N-acetylcysteine (NAC), vitamins C and E, Curcumin, and Livergel. Hemodialysis was performed for the patients, with one undergoing hemodialysis and hemoperfusion after ingesting 250 mL of PQ 20%. After a six-month follow-up, all surviving patients were in good health.Conclusion: Various factors, such as early admission after exposure, prompt gastrointestinal (GI) decontamination, corticosteroids with Curcumin and Livergel, antioxidants, hemodialysis, and hemoperfusion in one case may have contributed to the survival of patients with PQ poisoning in this study. However, individual vulnerability should also be considered a crucial factor requiring further investigation

Utilization of Mean Platelet Volume for Predicting Ischemic Heart Disease in Diabetic and Non-Diabetic Patients

Background: This study aimed to evaluate the relationship between mean platelet volume (MPV) and myocardial perfusion abnormalities in patients with and without Type 2 diabetes mellitus (DM) using myocardial perfusion scans. Materials and Methods: This cross-sectional study compared 49 patients with Type 2 DM without overt cardiovascular symptoms with 49 healthy controls. Both groups underwent myocardial perfusion scans at rest and under stress conditions. Risk factors were assessed and recorded using a special research-made questionnaire. A complete blood count and MPV results were obtained using the Sysmex - KX-21 system. Data were analyzed using SPSS, with a p-value below 0.05 considered statistically significant. Results: No significant differences were observed between the two groups in terms of Summed Stress Score (SSS), Summed Rest Score (SRS), Summed Difference Score (SDS), Ejection Fraction (EF), and End Systolic Volume (ESV). The only marked variance was a higher average platelet count in the control group. Regression analysis revealed that a one-unit increase in MPV correlated with a 0.46 average increase in SRS in the control group (CI: 0.08-0.83, β: 0.46). Conclusion: MPV may serve as a predictive marker for myocardial perfusion abnormalities, especially in individuals without diabetes. This simple metric could act as an early indicator for coronary artery disease

Deproteinization Process of Chitin from Dried Shrimp Shells (Litopenaeus vannamei) Using Papain and Nanochitin Characterizations

Background and Objective: Chemical treatments in chitin extraction from shrimp shell wastes have affected the environment. Shrimp shell primarily bonds chitin with inorganic salts, lipids, proteins and pigments. Extraction of chitin from shrimp shells involves protein separation processes. Deproteinization process of chitin from dried shrimp (Litopenaeus vannamei) shells with papain enzyme was optimized and nanochitin as a derivative product of chitin was characterized. Material and Methods: Effect of hydrolysis time, temperature and enzyme concentration were optimized using RSM Box-Behnken method to maximize chitin yields. Nanochitin was prepared using dialysis and ultrasonic methods and characterized for physical characteristics using scanning electron microscope, particle size analysis and Fourier transforms infrared spectroscopy. Results and Conclusion: Optimum conditions using enzymatic hydrolysis at 6 h, 50 oC and 1.25% papain decreased the protein content from 33.66 to 2.31% and produced a high chitin yield (46.03%). Deproteinization using enzymatic hydrolysis method was more efficient than that using fermentation. Data of scanning electron microscope, particle size analysis and Fourier transforms infrared spectroscopy showed that the characteristics of chitin and nanochitin products were similar to those of chemical treatments for chitin products. Conflict of interest: The authors declare no conflict of interest. Introduction   Litopenaeus vannamei is one of the shrimp species that includes high commercial values and produces abundant shrimp shell wastes. Production of shell wastes from crustaceans was predicted to be 3.14 million metric tons per year worldwide [1]. Hundreds of shellfish wastes are generated from seafood manufacturing and daily Asian consumption [2]. Shell wastes from crustaceans contain a high quantity of chitin, a polysaccharide material that is important in biological functions and is biodegradable and compatible. Chitin and its derivatives are used in various fields such as pharmaceutical, food, textile and waste water-treatment industries [3]. Chitin in the shrimp shells is bonded with majorly inorganic salts, calcium carbonate, proteins, lipids and pigments. Therefore, isolation of chitin from shrimp shells involves protein separation processes and mineral separation [4]. Structure of chitin is arranged with N-acetylated glucosamine and glucosamine units, linked by β(1,4) covalent bonds. Corresponding to this structure, chitin is stable to chemical and biological actions and the linkage of chitin is similar to the linkage of cellulose [5]. Generally, chitin is extracted through demin-eralization using acid treatment and deproteinization using alkali treatment. These treatments affect the environment and finding an alternative process that is more friendly to the environment is still necessary. Deproteinization process for chitin extraction from shrimp shells can be carried out via chemical, enzymatic and microbial processes [6, 2]. The chemical treatment involves mineral acid at high temperatures, resulting in high volumes of polluted waste containing mineral acids in the washing process. These treatments are harmful to the environment due to high concentrations of mineral acids [7]. Deproteinization with enzymes is a zero waste system resulting in high yields of chitin products. Protease hydrolyzes proteins in the matrix efficiently [8]. Commercially purified enzymes such as alcalase, papain, pepsin and trypsin have been used in chitin extraction studies to remove protein from crustacean shells [9].  Chitin is a biopolymer containing microfibrillar and semicrystalline structures. Based on data of the infrared (IR) spectra and X-ray crystallography (XRD), chitin is naturally in the forms of α-chitin, β-chitin and ɣ-chitin. Characteristics of chitin such as solubility, porosity and surface area restrict its uses. To solve this problem, various derivatives such as chitosan, chitin nanofibers and chitin nanowhiskers are produced [10]. Chitin nanofibers have been prepared via several methods such as ultrasonication, mechanical treatment, gelation and electrospinning [11]. Chitin nanofibers from species such as crabs, prawns and mushrooms have been prepared using mechanical and chemical treatments. The acidic medium was verified in the decrease of chitin nanofibers extracted from crab shells [12]. Under certain extraction conditions, chitin microfibrils are isolated in the form of nanocrystals and nanofibers. Their unique characteristics have been studied and used in food, cosmetics and medical industries [13]. Characteristics of chitin depend on the organisms and chitins may lay in α and β allomorphs shapes. These forms were assessed by the orientation of microfibrils that could be characterized using infrared, nuclear magnetic resonance (NMR) spectroscopy and XRD analysis [14]. Probiotic microorganisms have been studied for the demineralization treatment of crustacean shells. Chitin extraction using microorganisms was carried out simultaneously. Shrimp shells (Penaeus monodon) were fermented with lactic acid bacteria (LAB) and chitin was separated by adding carbohydrates [10]. Based on XRD and NMR data, chitin extraction via enzymatic process is an alternative method to preserve its native structure [1]. Shelma et al. reported the chitin nanofiber preparation via acid hydrolysis of the chitin powder followed by dialysis and ultrasonication [15]. Chitin from P. vannamae byproducts was prepared by associating enzymatic acid-alkaline strategies to achieve further sustainable processes [16]. Moreover, chitosan was produced through papain extract to help deproteinization process. Papain is achieved from the papaya plant with the endopeptidase, dipeptidase and exopeptidase activities. The optimum condition of this process was at 7 h of enzymatic hydrolysis and 25% of papain [8]. The current study was aimed to optimize deproteinization process of chitin from dried shrimp (L. vannamei) shells using low concentration papain (0.75–1.25%) and to achieve nanochitin, which was prepared via dialysis and ultrasonic methods. Furthermore, nanochitin products were characterized through physicochemical characteristics to verify their quality. Materials and Methods 2.1. Materials               Dried white-shrimp (L. vannamei) shells were provided as byproducts of a shrimp processing industry at Muara Gading City Bekasi, West Java, Indonesia. Commercial papain (CAS no. 2323.627-2) (Xian Arisun ChemParm, Shaanxi, China) was purchased in powder form. All chemicals used included laboratory grades. 2.2. Chitin extraction from the shrimp shells Chitin from the sample was extracted using method of Hongkulsup et al. [1] with some modification. The extraction was carried out at two steps, including demineralization and deproteinization. In demineralization process, shrimp shells were ground to achieve a size of 100 mesh. Shrimp shell powder was extracted using 1.5 M HCl (ratio 1:10, w/v) at 25 oC for 6 h and at 150 rpm. Mixture was filtered using vacuum filter and the residue was mixed with distilled water (DW) to achieve neutral pH. Then, residue was dried at 50 oC for 6 h. Dried residue was mixed with 0.75–1.25% w/v papain in a phosphate buffer pH 7 and heated at 40–50 oC for 3–6 h. Hydrolysis was stopped at 90 oC and set for 20 min. Mixture was filtered and the residue was mixed with DW until neutral pH was achieved. Then, residue was dried at 50 oC for 6 h. Total residue was assessed gravimetrically and the soluble proein content in the residue was analyzed using modified Lowry method. Briefly, 1 g of residue was diluted with DW up to 1 ml and filtered using Whatman filter papers. Then, 0.5 ml filtrate was mixed with 5.5 ml of alkaline CuSO4 reagent and incubated at room temperature (RT) for 10 min. Solution was mixed with 0.5 ml of folin phenol reagent. Then, sample solution was mixed with 3.5 ml of DW and the absorbance was measured at 650 nm. The protein soluble content was assessed by plotting bovine serum albumin (BSA) standard curve [17]. 2.3. Optimization of deproteinization of shrimp shells using Box-Behnken method                Optimum condition of the deproteinization process was predicted using response surface methodology (RSM)- Box-Behnken method. Optimization of deproteinization was carried out using three factors of effects of hydrolysis time, temperature and enzyme concentration (Table 1). Proportions of total residue, chitin and protein concentration were used as the responses data. Fifteen trials were carried out indiscriminately. The center value was chosen based on the references, which were 1% papain, 45 oC and 6 h [18, 19]. Design Expert 13.0 software was used in this study. 2.4. Assessment of chitin               Chitin content was assessed using adaptation of the Morrow method [20] with some modification. One gram of the sample was mixed with 40 ml of 1 M HCI and mixed at RT for 2 h. Chitin residue was separated using vacuum filter with a porous sintered glass disc and washed several times with water to reach a neutral pH. The residue was washed off and transferred into a beaker containing 40 ml of 5% NaOH and stirred at 100 °C for 2 h. Chitin product was separated using filter paper (Whatman no. 41, USA) and then rinsed with water until a neutral pH was achieved. Content of the chitin (%) was assessed gravimetrically. 2.5. Nanochitin preparation               The selected chitin sample, which was prepared at optimized conditions, was soaked in 3 M HCl for 90 min at 90  oC. Suspension was precipitated by centrifugation at 6000 rpm for 10 min. Nanochitin from the precipitated fraction was prepared for dialysis and ultrasonic treatments using Mincea method [11] with modifications. Suspension of chitin was transferred to a dialysis bag (cellulose membrane with cut-off proteins mol. wt ≥ 12,000) and dialyzed in DW by changing the water every 2 h for three times. Dialysis was carried out until pH 6 was reached. Ultrasonic treatment of the chitin sample was carried out at pulse of 1/1 and amplitude of 60% (750 W, 20 kHz) for 6 h to 0.1% (w/v) of the suspension. Based on the modification of Wu and Meredith method [21], these samples were freeze-dried at -60 oC for 10 h. 2.6. Microstructure identification               Microstructure of the freeze-dried samples was assessed using scanning electron microscope (SEM) (JSM-IT30, Jeol., Akhishima, Tokyo, Japan). These samples were put in a sample holder and layered with a thin layer of gold (±10 nm). Observation was carried out by accelerating voltage at 20 kV based on a previous method. 2.7. Particle size distribution               Particle size distribution of the samples was analyzed using particle size analyzer (Zetasizer Nano ZS Malvern,UK)  based on Shelma et al. method [15] with modifications. Sample was dispersed in Tween 80 (0,4%; w/v) with a ratio of 1:4. 2.8. Fourier transforms infrared spectroscopy (FTIR)               Spectra of the samples were analyzed using Fourier transforms infrared spectroscopy (FTIR 1000, Perkin-Elmer, USA) at mild conditions and method of KBr pellet scanning. Based on previous studies, KBr (100 mg) and the sample (1 mg) were mixed entirely until KBr pellet was formed. Then, samples were scanned at spectral ranges of 400, 4200 and 4200 cm-1. Results and Discussion 3.1. Optimization of deproteinization of the shrimp shells               The optimum conditions of the enzymatic hydrolysis in the deproteinization process of white shrimp shell powder were predicted using RSM. Fifteen trials were carried out based on the RSM-Box Behnken design. The Box–Behnken design (BBD) is a widely used RSM design that is useful for ascertaining cause-and-effect correlations between factors and responses in experiments. The BBD needs three levels and can be used for factors of 3–21 [22]. Hydrolysis factors and their responses are provided in Table 1. Data showed that the total residue of the products ranged 74.14–80.76%, chitin content ranged 41.52–49.06% and protein content ranged 2.31–6.82%. Analysis of variances (ANOVA) was calculated and p-values of the total residue, soluble protein and chitin content are present in Table 2. Papain concentration (C) and its interaction with temperature (AC) and hydrolysis time (BC) significantly (p < 0.05) affected the total residue of shrimp shell powder. The hydrolysis time (A) and its interaction with the papain concentration (AC) significantly (p < 0.05) affected the chitin content. However, p-values of the soluble protein contents showed that treatments were not significant (p > 0.05). The equation for estimating the optimal condition for all responses (Y1, Y2 and Y3) from the shrimp shells is present in Table 3. Total residue included the yield of the dried product after the deproteinization process with the papain enzyme. Chitin extraction via enzymatic hydrolysis needs removing proteins from the crustacean shells, minimizing the deacetylation and depolymerization processes. This process may be carried out before or after the demineralization step of solid materials for accessibility of the reactants. Efficiency of the enzymatic treatments is inferior to chemical methods ranging 5–10% of the residual protein attached to chitin [9]. Commercial enzymes such as alcalase, econase, pancreatin and other proteases were used in the chitin extraction of shrimp and crustacean shells. The objective of these treatments was to eliminate the protein contained in the waste of shells. Proportion of the chitin ranged 16.5–22% [7]. Combination of the chemical agents and enzymes has been studied to increase yields of the chitin products. Use of sodium sulfite and alcalase was the best treatment for protein recovery. Characteristics of the chitin sample were similar to those of the commercial food-grade products [6]. Three-dimensional (3D) response surfaces of the response; of which, one of the factors is fixed at the central point and the other is varied, are present in Figure 1. The highest predicted chitin content is indicated by the surface confined in the smallest ellipse in two-dimensional (2D) contour plots. This indication was correlated with the interaction between hydrolysis time and papain concentration significantly. This was similar to the results of ANOVA analysis (Table 2). The 2D contour plots showed effects of hydrolysis time (A) in the chitin content prediction (Fig. 1c). However, stagnation was observed in the chitin content with increasing temperature (Fig. 1b). To achieve the optimum condition of the deproteinization process, an optimization process was analyzed using Design Expert 13.0 RSM optimizer software. The three factors (time, temperature and papain concentration) were adjusted in the importance level 3 (+++) and responses (total residue, soluble protein and chitin yield) were adjusted in the importance level 5 (+++++). The optimum condition with desirability of 0.619 was observed for chitin extraction at 6 h, 50 oC and 1.25% papain. Further, all the responses of the products were validated through laboratory experiments. The experimental and predicted values are present in Table 4. Data showed that the experimental and predicted values were in the range (95% prediction interval); thus, reliability of the optimized condition was verified. The RSM-Box Behnken design was successfully used to assess effects of hydrolysis time, temperature and papain concentration on deproteinization process to produce higher chitin contents. Chitin from the molted shrimp shells was extracted using a chemical method. The optimum condition of deproteinization was achieved in 3% NaOH at 50 oC for 6 h with a residual protein content ≤ 1% [23]. Yulirohyami et al. (2024) reported that chitosan was prepared through processes, including depigmentation, demineralization, deproteiniza-tion and deacetylation. The optimum condition of depro-teinization process was reached at 25% of papain for 7 h of hydrolysis. This study showed that the hydrolysis time of chitin deproteinization affected deacetylation degrees of chitosan [8].               Proteases have been used for chitin extraction from shrimp byproducts. Residual proteins in shrimp wastes included 1.3 and 2.8% after treatment with chymotrypsin and papain enzymes. Combination of papain with other proteases that was used for deproteinization of shrimp wastes showed that the protein removal rates were low [24]. As an alternative to chemicals and decreasing shrimp wastes, 0.2% alcalase was shown to include activities in decreasing protein contents in shrimp wastes from 49.43 to 4.12% [25]. The enzymatic deproteinization of shrimp processing wastes has limited chitin yields nealy  4.4 to 7.9% of the total weight. This might be due to the residual of short peptides appropriately bonded to the compound of chitin. Use of combination agents with protease significantly decreased the protein fraction. Through this combination, protein fraction significantly decreased, assuming that protease degraded disulfide bonds of the shrimp head waste proteins that facilitated entry of sulfite ions [6]. Use of exoenzymes and proteolytic bacteria in deproteinization of demineralized shells produced liquid protein and solid chitin fractions [2]. Papain is a commercial enzyme, which includes endopeptidase, dipeptidase and exopeptidase activities. Binding affinity and catalytic efficiency of papain are affected by the substrate, temperature and incubation time [8]. 3.2 Physicochemical characterization of Chitin Characteristics of chitin, including degree of deacetylation, morphology and molecular mass, vary depending on the extraction method and origin of chitin [14]. For example, chitin achieved by the chemical extraction showed a tightly packed morphology, while a slightly microfibrillar structure was shown by chitins extracted via enzyme treatment [1]. Use of chitin increased significantly due to the prominent characteristics of its derivatives and nanostructure configuration, which are met for industrial processing. Techniques have been developed to produce chitin derivatives. For example, dialysis and ultrasonic methods  to produce nanochitin from shrimp shells; similar to those of the present study. Surface morphologies of the prepared chitin and nanochitin are present in Figure 2. Accordingly, porous-like honeycomb structure with no nanofibers on the surfaces was observed in the chitins (Figure 2a) and nanochitin achieved via dialysis process (Figure 2b). The only difference between these two products was in the pore size as the pore width of chitin (3 μm ±5) was smaller than that of nanochitin (5–15 μm). This result indicated that during the acid hydrolysis process of nanochitin preparation, the amorphous part of chitin was removed, leaving the crystalline side and leading to increases in pore size. The nanochitin generated from the ultrasonic technique (Figure 2c) showed a nanofibrillar structure with a diameter of nearly 160 nm. This reveals that hydrolysis with a strong acid followed by the ultrasonication treatment further facilitated dissolution process of the amorphous chitin [16]. Ultrasonication is a method to change the natural cithin into chitin nanofibers. Fibrillating chitin at 900–1000 W and 20 kHz in water (pH ±7) created nanofiber widths of 25–120 nm. High frequency of ultrasonication induced startling waves on the chitin surface that promoted their factorization with the axial way [26].  Chitin is naturally detected in crystalline microfibrils as a structural component, serving as a functional material that is needed by many organisms [14]. The pH of a solution in chitin treatment affects the surface morphology of chitin nanostructures, as a previous study demonstrated that the nanofiber structures of chitin were destroyed to small irregular shapes under high alkaline environments [23]. Furthermore, chitin nanofibrous structure formed due to chitin nanofibers are not soluble and result in versatile porous structures of the products by adjusting the freezing temperature. Freeze-drying technique includes the potential for the assembly of the nanofibrous structure of water-dispersible materials [21]. Particle size distribution is an important characteristic that affects functionality of the chitin products. The chitin sample included two peaks in the spectra, which were dissolved in 0.4% of Tween 80 solution (Fig. 3a). The Z-average of chitin from the shrimp shells was 511.7 nm and the highest intensity was 21.8%. Moreover, nanochitin samples showed three peaks with Z-averages of 101.7 (Fig. 3b) and 345.4 nm (Fig. 3c), respectively. Nanochitin produced via dialysis method showed a Z-average of the particles smaller than that produced by the ultrasonic method. However, the intensity of nanochitin products was still lower than that of untreated chitin samples. Particle size distribution of the chitin nanofibers demonstrated a bimodal curve with majority sizes of 20–300 nm [15]. In this study, additional peaks in nanochitin products were assumed as degraded chitin products. Temperature of the experiments affected number of the peaks in spectra. For higher temperatures, large particles were observed, which might be caused by degradation of the chitin particles. The lower temperature of ionic liquids was further favorable, resulting in a narrow particle range of particle size distribution spectra [27]. Ionic liquids could change the chitin structure, able to modify the particle size [28]. The FTIR spectrum of chitin is present in Figure 4. Chitin sample showed similar spectra with nanochitin, which was prepared via dialysis and ultrasonic methods. Spectra at 3258 and 2924 cm-1 were recognized as N-H and C-H stretching vibrations. The amide I band wa

Selenium-Enriched Microalgae: A Novel Bioactive Strategy for Immune System Enhancement

Background and Objective: Selenium-enriched microalgae represent a promising functional food and nutraceutical resource, offering a symbiotic blend of bioactive molecules and essential micronutrients. Selenium, a trace element, is vital for immune system control, antioxidant defense systems, and cell homeostasis. Microalgae possess high nutritional value and rapid growth capabilities, allowing them to bioaccumulate selenium in the form of organic compounds like selenomethionine and selenocysteine. These organic forms have increased bioavailability and reduced toxicity compared to inorganic selenium supplements. This review examines the latest advancements in the cultivation, selenium-enrichment strategies, and biological effects of selenium enriched microalgae, with a particular focus on their immunomodulatory properties. Results and Conclusion: The review highlights the potential applications of selenium enriched microalgae in disease prevention, immunotherapy, and functional food development. Also, the immunomodulatory effects of selenium enriched microalgae have been illustrated. Selenium and Se-enriched microalgae, due to their effective roles in enhancing immune function, reducing inflammation, and providing highly bioavailable forms of Se, hold strong potential for applications in food biotechnology and nutritional supplements. Despite existing challenges in optimizing production and clarifying mechanisms of action, the future outlook is highly promising given the growing demand for functional foods and natural health-promoting solutions. Keywords: Antioxidant, Bioavailability, Chlorella vulgaris, functional foods, Immunomodulation, Selenium-enriched microalgae, Selenoproteins, Spirulina, Sustainable biotechnology. Introduction   Selenium (Se) is an essential element for the proper functioning of immune system cells, including macro-phages, natural killer (NK) cells, neutrophils, and T lymphocytes. This element plays an essential role in reducing oxidative stress, inflammation, and preventing the spread of infectious diseases, especially when its serum concentration is adequately increased through the diet [1, 2]. It is involved in modulating immune and reproductive function. This micronutrient induces the production of selenoproteins, helping to protect cells against reactive oxygen species (ROS) [3]. Se primarily functions in the body as selenoproteins, which enhance the regulation of the human immune system in various ways. Immunity is one aspect of human health is influenced by Se levels and the expression of selenoproteins in the body [4]. Although the exact physiological role of Se is unknown, it has been determined that it exhibits most of its effects by being incorporated into selenoproteins. One such class of proteins is iodothyronine deiodinases, which are essential in the maturation of thyroid hormones T3 and T4. These hormones control body weight, development, and metabolism. A deficiency in Se may therefore disrupt the process of maturation of T3 and T4, leading to stunted growth. [5]. Dietary supplements containing l-selenometh-ionine are a source of Se for nutritional purposes that is easily absorbed and utilized by the body, approved for adult use at a dosage level of less than 250 mcg per day [6,7]. Seafood and meat are regarded as the primary sources of Se for humans, providing more than 70% of daily Se intake. In contrast, fruits and vegetables contribute only a small fraction to this consumption. Se is crucial because of its antioxidant and chemoprotective roles at low levels, offering protection against various cancers, heart diseases, and type 2 diabetes. Therefore, food systems must produce sufficient amounts of this essential trace element to ensure a daily intake of at least 40 micrograms, which supports the optimal expression of Se enzymes, and potentially up to 300 micrograms per day to lower cancer risk [8]. The availability of Se varies significantly depending on the geographical region. In some places, soil Se concen-trations are limited or reduced [9]. It is reported that approximately 1 billion people worldwide are affected by Se deficiency, meaning that diets are limited to products harvested from these regions or are nutritionally unbalanced and may result in Se deficiency [10]. The global population is rapidly increasing, leading to the emergence of new diseases and placing a significant strain on healthcare systems. Therefore, it is more important than ever to research immune modulators, including Se, to tune the immune system to help combat new pathogens in a changing world [11, 12]. Essential minerals for optimal health are provided by edible plants, which are collected from the environment, namely, soils and aquatic sediments. The consumption of Se-rich plants makes this element bioavailable to humans [9]. Soils used in agriculture can be challenging to meet daily Se requirements through diet if they are low in Se. Se mineral salts are mainly used in food supplements and animal feed, and this supplementation method with sodium selenite or selenate also has disadvantages [13-15]. Today, organic Se-enriched foods, mainly plants, animals, and microorganisms enriched with Se, have been widely developed to address the problem [13]. In recent years, the production of Se-enriched microalgae has attracted much interest as an efficient and accessible method for producing organic Se [16, 17]. Microscopic algae, or microalgae, grow in freshwater and marine environments and have the capability to convert sunlight energy into chemical form. The two most notable ones are Arthrospira platensis (Spirulina) and Chlorella vulgaris. Spirulina is blue-green algae and a nutritional powerhouse containing a plethora of vitamins and poly-unsaturated fatty acids and plays critical roles in nearly every activity of human life [18, 19]. Microalgae represent promising sources of bioactive compounds suitable for pharmaceutical and food uses [20]. Humans have been eating algae as a food crop and dietary supplement for thousands of years. Algae also serve as a natural carbon sequester, counteracting global warming and reducing the pressure on arable land and freshwater resources for conventional food crops. For the best nutritional com-position of algae, there is a need to stress their cultivation keeping in view many environmental parameters such as pH, intensity of light, availability of nutrients, availability of CO2, temperature, and mixing conditions [21]. Micro-algae, especially Chlorella vulgaris, are well-known as excellent sources of protein, balanced amino acids, and essential vitamins and micronutrients. Due to their high nutritional content, these microalgae are commonly con-sumed by humans. Chlorella vulgaris can absorb inorganic Se salts and convert them into protein-based compounds such as selenomethionine, selenocysteine, and methyl selenocysteine [16]. Thus, the aim of this review is to draw on scientific studies used to explore how Se-enriched microalgae modulate the immune system, providing updates on the recent advancements in this research area. This review is distinct from others due to its up-to-date and broad inclusion of all research drawn, thus giving the reader a com-prehensive overview of Se-enriched microalgae immune-modulatory effects. Types of Se-enriched microalgae Se-enriched biomass (Se-Chlorella) can serve as both a dietary supplement and an antioxidant [16]. Food products derived from Chlorella include green tea powder, soups, noodles, bread, biscuits, ice cream, and soy sauce. Raw Chlorella is commonly offered in tablets, capsules, powders, granules, and drinks. Noodles containing 1.5% Chlorella extract are exceptionally nutritious. Adding Chlorella powder to barley bread not only enhances its nutritional content but also improves its appearance and flavor. Additionally, Chlorella is used as a food additive to enhance the taste and quality of pasta, wine, and fermented foods. With vitamins C, K, A, and E, Chlorella is valuable for pharmaceuticals, animal feed, food additives, aquaculture, and cosmetics [22]. Chlorella can develop under conditions of light, carbon dioxide, water, and a minimum amount of nutrients, thus its culture is easy. The life cycle of the microalga is simple while its metabolic processes are as complicated as those of superior plants, which makes it capable of synthesizing high amounts of proteins, carotenoids, vitamins, and minerals. Being so, it is a popular source of nutritious food. Among numerous species of Chlorella, Chlorella vulgaris, Chlorella sp., and Chlorella pyrenoidosa are the most utilized in industrial production and scientific research [23]. Se exerts its biological effects via selenoproteins. Various microalgae, such as Chlamydomonas, Volvox, Ostreococcus, Micromonas, and Emiliania, have been identified as possessing selenoproteins [24]. Spirulina, a type of microalgae, possesses antioxidant properties and can be fortified with Se during its growth process. Research revealed a hierarchy in the distribution of Se and the expression of selenoproteins based on the form of supplementation. Supplementing with sodium selenite enhanced glutathione peroxidases (GPx) activities and selenoprotein expression, whereas Se-enriched spirulina was more effective in restoring Se levels [5]. In 1992, the World Health Organization listed it as a future food. Spirulina is used as a dietary supplement due to its high protein value, vitamins, β-carotene, and phycocyanin-like pigments. It contains anti-inflammatory, anti-cancer, and antioxidant properties. In vivo and in vitro studies have demonstrated that spirulina supplementation can reduce markers of oxidative stress and enhance the activity of antioxidant enzymes. Also, as it matures, spirulina may be enriched with elements like Se, which is incorporated into organic molecules like selenomethionine and seleno-cysteine [5]. In recent years, although there has been a notable rise in algal production, it remains insufficient to satisfy the high commercial demand for biomass [21]. The dried biomass of microalgae is an excellent option for food use because of its rich nutritional content, economical processing expenses, and various health advantages [25]. Table 1 provides a summary of the types of Se-enriched microalgae. Biotechnological ethods for seleni-um enrichment (cultivation methods and genetic modifications) The ideal harvesting process for microalgae should maintain the integrity of the composition of the biomass and facilitate effective recovery of the target product. Traditional harvesting processes involve filtration, centrifugation, flotation, flocculation, electroflocculation, sedimentation, electrolytic treatment, electrophoresis, and magnetic separation. Incorporating an additional step of chemical or biological coagulation or flocculation into these processes can enhance the efficiency of the process as well as reduce operating expenses. Centrifugation is widely utilized for microalgae harvesting due to its effectiveness and speed in cell recovery. Centrifugation involves centrifugal force to separate microalgal biomass from the culture medium, from which excess water may easily be drained. Centrifugation can achieve up to 98% yield, but its greatest flaws are high energy demand and the potential for cellular structure breakdown during the process [23]. Microalgae are highly efficient at fixing carbon dioxide during their growth, so they do not require arable land for cultivation. Microalgae biomass production involves three main steps: cultivation, harvesting, and processing. Among these, harvesting is particularly challenging due to its high costs, and it has a direct impact on the processing stage [26]. The harvesting stage of microalgae is crucial for its overall production. Research has shown that harvesting accounts for 20–30% of the total production cost. Key challenges in the harvesting and dewatering process include the small size of the cells (<30 µm), their low concentration and dilute presence in the culture medium (<1 g.l-1), the highly electronegative nature of the cell membrane surface, and the relatively fast growth rate of the algae. As a result, the energy required for the harvesting process exceeds the energy content of the microalgal biomass itself [26]. Coagulation using polyaluminum chloride (Al2O3) and hydrophilic polytetrafluoroethylene membranes has been used as a sustainable technology for harvesting microalgae. Transparent exopolymer particles that are also produced by microalgae have been used to reduce the fouling of membranes in microfiltration. The particles prevent membrane fouling when solutions are collected through the use of the filtration-based method, hence improving filtration efficiency [26]. Microalgae possess the ability to absorb inorganic Se and combine it with amino acids to create Se amino acids, including selenomethionine, selenocystine, which are advantageous for the health of both humans and animals. Furthermore, Se can be added from a primary source to cultivate Se-enriched microalgae in domestic wastewater, which can act as a nutrient base for microalgae growth. Low levels of Se may stimulate microalgae growth. Turbidity significantly increased over the incubation period, indicating that microalgae growing in domestic wastewater treatment systems can tolerate such high Se concentrations. The removal process facilitated by microalgae in HRAP involves biomass assimilation, as microalgae can use these substances to produce cellular components, including proteins, nucleic acids, and carbohydrates [27]. A post talks about various methods of cultivation, starting with open systems using open ponds for growing microalgae. They rely on natural water and sunlight but are prone to environmental stresses and the risk of contamination. It also talks about closed systems, which use photobioreactors or bioreactors that yield closed environ-ments for the growth of microalgae. These closed systems are efficient and less likely to cause contamination compared to open operations. In addition to this, advanced techniques like mutagenesis and genetic alteration are also discussed, which are used in order to enhance some characteristics of microalgae to maximize biomass production and quality. The article also emphasizes the necessity of specific environmental conditions, such as pH, temperature, and light intensity, in order for maximum growth and production. Altering these can lead to increased production of Se and other secondary products. Lastly, nutrient-rich conditions are suggested as an alternate approach to enhance microalgae production. Nevertheless, it is essential to maintain adequate levels of nutrients so as not to compete with Se absorption. These culture processes enhance microalgae productivity across various industries [16]. One advantage of cultivating microalgae in a mixotrophic manner is the increased biomass production [28]. Today, with advancements in microalgae harvesting, methods such as bioflocculation, electroflocculation, bio-electroflocculation, ultrasound/hydrodynamic techniques, magnetic nanoparticle flocculation, and phototaxis-based harvesting are employed [29]. Table 2 shows biotechnological methods for Se enrichment. Analysis of bioavailability of selenium in microalgae The way Se is absorbed differs among animal species and is affected by various factors, including physiological traits, functional status, the number of intestinal contents, the chemical forms of Se, the duration of Se’s stay in the intestine, and the methods of Se administration. Se is mainly found in the liver, kidneys, heart, and pancreas, with muscles, bones, and blood having the following highest levels, while fat tissues contain the least amount. Typically, in normal circumstances, the Se that animals metabolize is mainly eliminated through urine and feces. However, when Se is consumed in excessive amounts, breathing also becomes a significant pathway for its excretion. Moreover, Se can be removed from the body through hair and sweat [30]. The bioavailability of Se in microalgae, indicating how much Se can be absorbed in the gastrointestinal tract, was assessed in a study. Grinding the microalgae, their cell walls are broken down, facilitating the release of Se from the biomass during digestion. Under gastrointestinal conditions, 49% of Se from raw microalgae and 63% from ground Se-enriched microalgae were solubilized, indicating their potential bioavailability. A similar level of Se bio-availability (approximately 49%) has been observed in Se-enriched Chlorella vulgaris. Additionally, it's important to highlight that the bioavailability of Se in Se-rich yeast widely recognized as a leading organic Se supplement is generally greater than that found in microalgae cultivated in HRAP-Se. Furthermore, the type of Se also influences absorption rates [27]. The Se in food sources is mainly in organic forms, including selenocysteine and selenomethionine, which are more easily absorbed than inorganic forms like selenite or selenate. There is a significant risk of losing dietary Se during processing and cooking; for instance, the refining of grains can diminish Se levels by 50 to 75%, while boiling can lead to an additional reduction of 45%. Microalgae that utilize Sec, such as Nannochloropsis oceania, can enhance the organic Se content and can thus be used as Se-enriched food or feed additives [24]. Organic Se compounds have higher bioavailability and usability than their inorganic counterparts. Studies also indicate that Se-enriched plant products usually provide greater bioavailability than animal-based products. Moreover, adding Se to proteins can not only increase their Se content but also improve their functional qualities, including emulsion stability and gelling properties, along with enhancing their bioavailability [31]. In numerous microalgal species, selenite may exhibit greater toxicity compared to selenate, while the opposite can be true for other species. Due to its high solubility, selenate is more readily bioavailable to aquatic organisms than selenite, indicating that selenate might be the predominant dissolved form. Prior research has shown that selenate does not pose toxicity risks for some microalgal species [8]. Applications in food biotechnology development and utilization of function-al foods, fortified foods, beverages, and dietary supplements Microalgae cultivation is one factor that can enhance world food security and reduce the environmental impact of rising agricultural food production. The quality and safety of microalgae product, in turn, are both a function of proper cultivation, harvesting, and processing protocols since foreign organic and inorganic material occur as a result of environmental contamination or process operations when dealing with microalgae biomass [28]. Functional food production with microalgal biomass will see growth in the coming years as a result of growing demand for healthy foods globally. Microalgae incor-poration in food products is the next wave of evolution of the functional food industry, given that there is continuous need to search for new raw materials for use in creating new foodstuffs. Nevertheless, some of the major questions need to be answered to facilitate the successful development of microalgal-derived food products. Some of the other questions that belong to this category are how microalgal biomass interacts with the other components of the food matrix, the effect of microalgal addition on the sensory attributes of food, particularly color and flavor, textural and rheological modification of food products with microalgal addition, and changes and stability of microalgal constituents under various processing conditions [32]. Understanding the mechanisms and potential for Se accumulation in plants and animals is one of the most critical directions in developing nutritional science and supplementation [33]. To create algae-based food products and by-products, the first essential step is enhancing our knowledge of the biochemical makeup and digestibility of microalgae. In line with this, it’s important to assess more microalgal species for novel food approval, ultimately supporting the broader inclusion of these microorganisms in human diets over time [34]. Numerous studies have demonstrated that microalgae species, particularly from the Chlorella and Spirulina genera, are well-suited for direct use in their natural forms. This adaptability has led to their widespread application in the food industry, where they enhance the nutritional content of products like noodles, cookies, energy bars, and juices [23]. In the food and beverage industry, as well as within the cosmetics industry, the demand for spirulina has risen due to its fat-reducing, antioxidant, and anti-inflammatory properties [35]. An article discusses the production of beverages such as "selenized kawas," made from fennel grains soaked and germinated in solutions containing Na₂SeO₃. These beverages are considered a source of Se. Regarding Se in beer, yeasts (S. cerevisiae) can convert inorganic Se (Na₂SeO₃) into bioactive organic forms that are utilized in selenized beers. This process enhances the Se content in the beverages [36]. In a study, Chlorella vulgaris was blended into spreadable processed cheese, leading to increased magnesium, potassium, Se, Zn, iron, and antioxidant capacity, with greater enhancements observed at higher formulation levels. Microalgae were also incorporated into various snacks, breads, isotonic drinks, and yogurts; however, these products did not attract consumer interest due to their unappealing green color and distinct flavor. On the other hand, a dehydrated soup prepared with S.

Proteomics in Chronic Prostatitis: Biomarker Discovery, Molecular Pathways, and Emerging Targets for Precision Medicine

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a common and disabling urological disorder that affects quality of life in men. Despite accounting for most prostatitis cases, its causes remain unclear, involving immune dysregulation, oxidative stress, microbial factors, and epithelial barrier dysfunction. This uncertainty complicates diagnosis and treatment.Proteomics offers a high-throughput approach to identify proteins and pathways involved in CP/CPPS. Recent studies have profiled urine, seminal plasma, prostatic secretions, and serum to uncover biomarkers linked to inflammation, oxidative stress, and microbial virulence. These findings provide insights into molecular endotypes, guide new classifications, and point toward novel therapeutic targets such as cytokine signaling, pyroptosis pathways, and heat shock proteins. Although technical variability and small cohort sizes remain major challenges, integration of proteomics with multi-omics platforms and explainable AI may transform CP/CPPS management by enabling personalized diagnostics and targeted interventions

Investigating the effect of 8 weeks of aerobic training and Borage extract on the serum concentration of liver enzymes and lipid profile in rats with non-alcoholic fatty liver disease (NAFLD)

Objective: Non-alcoholic fatty liver disease is known as one of the chronic liver diseases that is closely related to obesity and metabolic disorders, and borage improves oxidative indices due to its antioxidant properties. The aim of this study is to investigate the effect of a part of aerobic exercise with borage extract on the serum concentration of liver enzymes and lipid profile in rats with non-alcoholic fatty liver disease. Materials and Methods: In this experimental study, 40 male Wistar rats weighing 250-300 grams were divided into 4 groups (N: 10). The groups were: Control group (suffering from fatty liver and receiving enough water and food/without exercise and supplements) supplement group (suffering from fatty liver and receiving 200 mg/kg borage extract) exercise group (suffering from fatty liver and doing aerobic exercise daily for 8 weeks) exercise + supplement group (suffering from fatty liver and receiving 200 mg/kg borage supplement and performing aerobic exercise for 8 weeks). At the end, blood was drawn from all the animals, then the serum lipid profile, including total cholesterol and triglycerides, as well as liver enzymes such as alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured by ELISA method. They were analyzed with SPSS and a significance level of <0.05. Results: The findings of the present study showed that in the comparison of total cholesterol serum level changes during the study period after poisoning, there was a significant decrease in the control groups (P=0.004) and supplement (borage extract) + exercise (0.047). P=0) was observed. Nevertheless, the training and supplementation groups showed a significant increase in serum total cholesterol levels during the study period. In addition, comparing the changes in serum triglyceride levels during the study period after poisoning, only a significant decrease was observed in the supplemented group (P=0.002). Also, comparing the changes of serum ALT level during the study period after poisoning, a significant increase in serum ALT level was observed in the control (P=0.045) and exercise (P=0.002) groups. The supplement group also showed a significant decrease in ALT serum level (P<0.001). In comparing the changes of AST serum level during the study period after poisoning, a significant decrease was observed in the control (P=0.003) and supplement (P=0.006) groups. Conclusion: The findings of the present study showed that 8 weeks of supplementing with borage extract alone reduced serum triglyceride, AST, and ALT levels, and 8 weeks of aerobic exercise with borage extract reduced total cholesterol levels in rats with non-alcoholic fatty liver disease. &nbsp

Ethical Considerations of Using Artificial Intelligence in Libraries at Medical Universities

Background: Artificial intelligence (AI) is increasingly integrated into medical university libraries, transforming information retrieval, knowledge organization, and user support. While AI offers efficiency and personalization, its adoption raises critical ethical concerns that align with both medical ethics and library science. Objective: This narrative review examines the ethical implications of AI in medical university libraries, focusing on four guiding principles: privacy and data protection, transparency and accountability, equity and access, and trust and user autonomy. Methods: A systematic search of the literature identified 43 relevant studies published between 2019 and 2025. Evidence was synthesized to highlight risks, ethical implications, and strategies proposed for responsible AI integration. Results: Privacy concerns centered on risks of data breaches, vendor misuse, and long-term data retention, requiring compliance with GDPR/HIPAA and adoption of encryption and anonymization protocols. Transparency and accountability challenges stemmed from algorithmic opacity and bias, necessitating audits, explainable AI, and shared governance. Equity and access issues reflected institutional disparities and barriers for digitally marginalized users, emphasizing open-source tools, multilingual support, and digital literacy programs. Finally, trust and user autonomy were threatened by over-reliance on automated systems, highlighting the need for librarian oversight, ethics education, and user feedback mechanisms. Conclusion: AI adoption in medical university libraries requires a robust ethical framework to safeguard privacy, promote transparency, ensure equitable access, and preserve user trust. Future research should focus on empirical evaluations, cross-cultural perspectives, and policy frameworks tailored to academic medical libraries

Smart marketing and advanced personalization in healthcare services: the role of consumer data in improving patient experience and brand interaction

Background: With the expansion of digital technologies in healthcare, smart marketing has emerged as a data-driven approach that plays a prominent role in enhancing patient experience and improving interaction with healthcare service brands. Methods: This descriptive-analytical study was conducted in a field setting. The study population consisted of 220 managers and experts in healthcare marketing from healthcare institutions and health-focused companies. Using Cochran's formula, a simple random sample of 140 participants was selected. The data collection tool was a researcher-developed questionnaire, whose validity was confirmed by digital health experts and reliability was verified with a Cronbach's alpha of 0.87. Data analysis was performed using SPSS software and multiple regression analysis. Results: The Results showed that smart marketing significantly and positively influences patient (customer) experience, explaining approximately 62% of its variance (β = 0.55, p < 0.01). Additionally, brand interaction contributed to 56% of the changes in patient experience (β = 0.44, p < 0.01), while the use of consumer data accounted for 22% of its variation (β = 0.81, p < 0.01). Furthermore, patient experience itself explained 28% of the variance in advanced Personalization, confirming its mediating role in enhancing personalize healthcare delivery. Conclusion: Data-driven smart marketing in healthcare can facilitate the design of personalized healthcare services, leading to sustainable patient interactions and gaining a competitive advantage in the healthcare market

Anxiolytic and Potential Neurotoxic Effects of Salvia hypoleuca in Mice

Anxiety disorders are prevalent worldwide, significantly impacting various aspects of patients’ lives. The use of Salvia species in ethnobotanical medicine has been well documented, with applications as diuretics, analgesics, anti-hyperhidrosis agents, laxatives, antipyretics, and antitussives. Among these, Salvia hypoleuca, an endemic plant in Iran, is recognized for its tonic, carminative, digestive, antispasmodic, anti-inflammatory, antioxidant, antimicrobial, and anti-nociceptive properties. The objectives of this study were to assess the anxiolytic effects of S. hypoleuca ethanolic extract and its potential neurotoxicity using established pharmacological models in mice. Animals were randomly allocated into five groups: one control group, three groups underwent treatment that received oral doses of S. hypoleuca at 30, 100, and 300 mg/kg, and a positive control group given diazepam at 10 mg/kg. Behavioral evaluations were performed using the light/dark test (LDT) and the elevated plus maze (EPM) tests. To evaluate potential neurotoxic effects, open-field and rotarod tests were also performed. The results specified that S. hypoleuca extract at doses of 30 and 100 mg/kg significantly enhanced entries and time spent in the open arms of the EPM, suggesting anxiolytic effects. In the LDT, the 30 mg/kg dose notably increased the time spent in the light box. However, the rotarod test showed a slight decrease in latency to fall at both 30 and 300 mg/kg doses, indicating possible motor impairment at higher concentrations. Open-field analysis revealed significant reductions in total distance moved and velocity at 30 and 300 mg/kg doses, suggesting potential locomotor suppression. Additionally, the total phenolic and total flavonoid contents of S. hypoleuca ethanolic extract were measured as 47.26 ± 2.87mg GAE/g DW and 31.73 ± 5.38 mgRE/g DW, respectively. In conclusion, our efforts suggest that oral administration of S. hypoleuca extract exhibits anxiolytic effects in mice, as demonstrated by improved performance in the EPM and LDT. However, the observed locomotor impairment at higher doses warrants further investigation to determine the optimal therapeutic dose and potential safety concerns

Magnetic resonance spectroscopy and diffusion contrast imaging in differentiating recurrence of primary brain tumors

Objective: Differentiation of brain tumor recurrence from necrosis caused by radiation after radiotherapy is often considered a radiological ambiguity. The purpose of this study is to investigate the effectiveness of magnetic resonance spectroscopy in differentiating the recurrence of primary brain tumors (glioma) from necrosis caused by radiation. Materials and Methods: 15 patients (with an average age of 40.07, 8 men and seven women) were examined in this study. The samples of this study were selected from patients who have already been confirmed to have a glioma brain tumor and requested radiotherapy. 12 weeks (or three months) after radiotherapy, MRS imaging sequences with CSI technique and DWI imaging sequence with b-value=1000 s/mm2 were performed for all patients, and metabolic coefficients and quantitative ADC maps were also obtained. Then, the average ADC value and metabolic ratios were calculated for all patients. Results: The pathological and clinical results of the patients were compared with the results of MRS and DWI. By analyzing the ROC curve on the data obtained from the two techniques implemented in this study, it shows that the area under the ROC curve for ADC maps was higher than that of the MRS technique, which shows that the results of DWI can reliably diagnose and differentiate tumor recurrence from radiotherapy necrosis. Also, the result of this study showed that tumor recurrence has a significantly lower ADC compared to radiotherapy necrosis (p=0.035). Conclusion: The results of MRS indicates that the metabolic ratios of Cho/NAA and Cho/Cr are higher in people who have had tumor recurrence than in people in whom necrosis from radiotherapy has been reported. DWI images obtained with b-value = 1000 s/mm2 provide more reliable results than MRS images with the CSI technique and are considered a valuable tool in differentiating tumor recurrence from radiotherapy necrosis; the reliability of both imaging results increases, and the ability to detect tumor recurrence from necrosis caused by radiotherap