rss_2.0The EuroBiotech Journal FeedSciendo RSS Feed for The EuroBiotech Journal EuroBiotech Journal 's Cover hydroperoxyl antiradical activity of Trolox in water: The effects of pH values on rate constants<abstract> <title style='display:none'>Abstract</title> <p>Trolox (6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid) is a highly hydrophilic α-tocopherol analog that is widely used as a standard against which the antioxidant ability of other chemicals is measured and represented in Trolox equivalents. However, the effect of pH values on the hydroxyl radical scavenging activity has not been fully studied yet. In this study, the HOO<sup>•</sup> antiradical activity of Trolox in water was studied. It was found that the H-abstraction of the O1-H bond determined the activity of the neutral and monoanion states, whereas the electron transfer reaction of the hydroxyl anion state determined the activity of the dianion state. Although the total rate constant increased following the increase in pH levels, the overall rate constant of the Trolox + HOO<sup>•</sup> reaction in water changed when pH levels rose due to the decrease in HOO<sup>•</sup> molar fraction. The results also revealed that at pH &lt; 2, the O1-radical was the main intermediate of the Trolox + HOO<sup>•</sup> reaction in water, whereas, at pH ---gt--- 5, the anion-radical was the significant intermediate. Thus the rate constants and the reaction intermediates vary with the pH values.</p> </abstract>ARTICLE2022-04-30T00:00:00.000+00:00Polyhydroxyalkanoate (PHA) Biopolyesters - Emerging and Major Products of Industrial Biotechnology<abstract> <title style='display:none'>Abstract</title> <p><bold>Background:</bold> Industrial Biotechnology (“White Biotechnology”) is the large-scale production of materials and chemicals using renewable raw materials along with biocatalysts like enzymes derived from microorganisms or by using microorganisms themselves (“whole cell biocatalysis”). While the production of ethanol has existed for several millennia and can be considered a product of Industrial Biotechnology, the application of complex and engineered biocatalysts to produce industrial scale products with acceptable economics is only a few decades old. Bioethanol as fuel, lactic acid as food and PolyHydroxyAlkanoates (PHA) as a processible material are some examples of products derived from Industrial Biotechnology.</p> <p><bold>Purpose and Scope:</bold> Industrial Biotechnology is the sector of biotechnology that holds the most promise in reducing our dependence on fossil fuels and mitigating environmental degradation caused by pollution, since all products that are made today from fossil carbon feedstocks could be manufactured using Industrial Biotechnology – renewable carbon feedstocks and biocatalysts. To match the economics of fossil-based bulk products, Industrial Biotechnology-based processes must be sufficiently robust. This aspect continues to evolve with increased technological capabilities to engineer biocatalysts (including microorganisms) and the decreasing relative price difference between renewable and fossil carbon feedstocks. While there have been major successes in manufacturing products from Industrial Biotechnology, challenges exist, although its promise is real. Here, PHA biopolymers are a class of product that is fulfilling this promise.</p> <p><bold>Summary and Conclusion:</bold> The authors illustrate the benefits and challenges of Industrial Biotechnology, the circularity and sustainability of such processes, its role in reducing supply chain issues, and alleviating societal problems like poverty and hunger. With increasing awareness among the general public and policy makers of the dangers posed by climate change, pollution and persistent societal issues, Industrial Biotechnology holds the promise of solving these major problems and is poised for a transformative upswing in the manufacture of bulk chemicals and materials from renewable feedstocks and biocatalysts.</p> </abstract>ARTICLE2022-04-30T00:00:00.000+00:00Cell contractile force measured using a deformable hollow capsule<abstract> <title style='display:none'>Abstract</title> <p>There are several possible ways to measure the contraction of cells <italic>in vitro</italic>. Here, we report measurements of the contractile properties of 3T3-L1 cells grown to confluence on 3D hollow capsules. The capsules were fabricated using the layer-by-layer polyelectrolyte deposition technique on a polymer core. After the polyelectrolyte film was completed, the core was dissolved to leave the hollow capsule. The contractile force of the cells was determined from the deformation in the capsule size induced by interruption of the actin cytoskeleton of the cells that adhered to the outer surface of the hollow capsules, using prior measurements of the elastic modulus of the capsule. From the measurements of the compressive modulus for the capsules (of 6.52 μN), those capsule deformations indicate that the forskolin relaxed the layer of cells by 19.6 μN and the cytochalasin-D relaxed the layer of cells by 45.6 μN. The density of cells in the layer indicated that the force associated with the forskolin-induced relaxation of a single cell is 3.2 nN and the force associated with the cytochalasin-D-induced relaxation of a single cell is 7.5 nN. The mechanism of action of forskolin through second messenger pathways to disrupt the assembly of actin stress fibres also explains its reduced effect on cell contraction compared to that for cytochalasin-D, which is a compound that directly inhibits the polymerization of F-actin filaments.</p> </abstract>ARTICLE2022-04-30T00:00:00.000+00:00Genetically modified mice for research on human diseases: A triumph for Biotechnology or a work in progress?<abstract> <title style='display:none'>Abstract</title> <p>Genetically modified mice are engineered as models for human diseases. These mouse models include inbred strains, mutants, gene knockouts, gene knockins, and ‘humanized’ mice. Each mouse model is engineered to mimic a specific disease based on a theory of the genetic basis of that disease. For example, to test the amyloid theory of Alzheimer’s disease, mice with amyloid precursor protein genes are engineered, and to test the tau theory, mice with tau genes are engineered. This paper discusses the importance of mouse models in basic research, drug discovery, and translational research, and examines the question of how to define the “best” mouse model of a disease. The critiques of animal models and the caveats in translating the results from animal models to the treatment of human disease are discussed. Since many diseases are heritable, multigenic, age-related and experience-dependent, resulting from multiple gene-gene and gene-environment interactions, it will be essential to develop mouse models that reflect these genetic, epigenetic and environmental factors from a developmental perspective. Such models would provide further insight into disease emergence, progression and the ability to model two-hit and multi-hit theories of disease. The summary examines the biotechnology for creating genetically modified mice which reflect these factors and how they might be used to discover new treatments for complex human diseases such as cancers, neurodevelopmental and neurodegenerative diseases.</p> </abstract>ARTICLE2022-04-30T00:00:00.000+00:00Serum collected from rats with myocardial infarction increases extracellular matrix accumulation by myofibroblasts isolated from myocardial infarction scar<abstract> <title style='display:none'>Abstract</title> <p>The effect on extracellular matrix content is believed to be an average of several serum derived compounds acting in opposition. The aim of the study is to determine whether whole serum of rats with myocardial infarction may modify the accumulation of extracellular matrix in cultures of myofibroblasts isolated from the myocardial infarction scar. A second aim is to determine whether the tested serum can also degranulate the mast cells. Serum was collected from rats with sham myocardial infarction, rats with myocardial infarction induced by coronary artery ligation and control animals. The experiments were carried out on myocardial infarction scar myofibroblasts or mast cells from the peritoneal cavity. The cultures were divided into three groups containing eight cultures each: one treated with serum from control rats, from animals after sham operation or from those after myocardial infarction. In all groups, the serum was used at concentrations of 10%, 20% or 30%. The total collagen content (Woesner method) glycosaminoglycan level (Farandale method), cell proliferation (BrdU), histamine secretion from mast cells (spectrofluorymetry), β1 integrin and α-smooth muscle actin expression (flow cytometry) were evaluated. Isolated cells were α-smooth muscle actin positive and identified as myofibroblasts. Serum derived from rats with myocardial infarction increased collagen and glycosaminoglycan content in the cultures and modified myofibroblast proliferation in a concentration-dependent manner. The serum also results in an imbalance between collagen and glycosaminoglycan levels. The content of β1 integrin was not influenced by myocardial infarction serum. The serum of rats with myocardial infarction is involved in regulation of collagen and glycosaminoglycan content in myofibroblast cultures, as well as the modification of their proliferation. These changes were not accompanied with integrin β1 density variations. The serum of the myocardial infarction rats did not influence the mast cell degranulation.</p> </abstract>ARTICLE2022-01-24T00:00:00.000+00:00SARS-CoV-2 Alpha Variant Infection of a Patient Immunized by Inactive Sinovac (CoronaVac) Vaccine<abstract> <title style='display:none'>Abstract</title> <p>Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first detected in December 2019, and shortly after pandemic has been declared by the World Health Organization (WHO) due to its unstoppable global spread. Considerable amount of effort has beenput around the World in order to develop a safe and effective vaccine against SARS-CoV-2. Inactivated and RNA vaccines have already passed phase three studies showing sufficient efficacy and safety, respectively. Nowadays, there is a noticeable dominance of SARS-CoV-2 variants with various mutations over the wild type SARS-CoV-2. However, there is no report showing the efficacy of these vaccines on these variants. This case study describes a thirty-eight-year-old male reported to be infected with SARS-CoV-2 alpha variant following two doses of inactive CoronaVac administration with a protective level of SARS-CoV-2 specific antibodies. The variant analysis of the virus reported to be positive for N501Y mutation.This is the first case in the literature demonstrating that inactive SARS-CoV-2 vaccine might have a lower efficacy on alpha variant.</p> </abstract>ARTICLE2022-01-24T00:00:00.000+00:00Chemical toxicity assessment and Physiological investigation in rats exposed to pyrethroid insecticide type 1 and possible mitigation of propolis<abstract> <title style='display:none'>Abstract</title> <p>The current investigation aims to study the potential protective effects of propolis methanolic extract (100 mg/kg BW) on the systemic toxic effects after dietary exposure concentration (1/100 LD50 for 30 days) of permethrin (PM) administered in experimental rats. In this experiment, we added propolis four weeks after PM -administration to examining the medicinal effects. Therapeutic use of propolis mitigated PM -induced deterioration of liver and kidney functions and myocardial damage measured by cardiac enzymes lactate dehydrogenase (LDH) and creatine kinase MB (CK-MB) in serum. In addition, propolis treatment (prophylactic and therapeutic) prevented PM-induced apoptosis index, including B-cell lymphoma protein 2 (BCL-2)-associated X (BAX) protein activates, and lipid peroxide (LP). The results showed propolis induced a significant decrease in serum levels of thyroid hormones (T3 and T4), proinflammatory cytokines tumor necrosis factor-alpha (TNF-α), interferon-gamma (INF-γ), interleukin one beta (IL-1β), interleukin 12 (IL-12), and interleukin 6 (IL-6). Besides, nuclear factor-kappa B (NF-kB), acetylcholine esterase (AChE), and hematological constituents. Cardiac biomarkers, liver, and kidney functions were substantially lower in propolis treatment. High-performance liquid chromatography (HPLC) and Gas chromatography–mass spectrometry (GC- MS) of the propolis-MeOH extract showed valuable antioxidant phenolics and flavonoids capable of alleviating oxidative stress through the free-radical scavenging efficacy and regulating signaling pathways of proinflammatory cytokines.</p> </abstract>ARTICLE2022-01-24T00:00:00.000+00:00Successful Mestrual Regularity and Spontaneous Pregnancies with a Resveratrol-Based Multivitamin Supplement in Women with Idiopathic Premature Ovarian Insufficiency<abstract> <title style='display:none'>Abstract</title> <p>Premature ovarian insufficiency (POI) is a clinical syndrome defined by loss of ovarian activity before the age of 40 years and is characterized by menstrual disturbance, follicle stimulating hormone (FSH) concentration above 40 IU/l and infertility. In some patients the best option is to conceive spontaneously since many treatment strategies remain unsuccessful or involve eggs donation. In this case report series, we describe the effects of a resveratrol-based multivitamin supplement containing trans-resveratrol, folic acid, vitamin B6, B12 and D, in six women with poor prognosis of pregnancy due to POI and evaluate the achievement of desired conception.</p> <p>These women, aged less then 40 years, suffered from menstrual irregularities, anovulation and infertility. They all had normal karyotype, and no history of ovarian surgery, radiation exposure or chemotherapy. Blood test showed at least two values of FSH above 40 IU/l.</p> <p>Four out of six patients with POI conceived after 3-6 months of a resveratrol-based multivitamin supplement, ultimately giving birth to a full-term baby. Regular menstrual cycle was restored in all patients after two to four months the start of treatment. In conclusion the treatment with a resveratrol-based supplement improved menstrual regularity and suggest a useful potential of this supplementation in some cases of POI.</p> </abstract>ARTICLE2022-01-24T00:00:00.000+00:00Aerobic Composting of Sugar Pressmud with Stabilized Spentwash and selected Microbial Consortium<abstract> <title style='display:none'>Abstract</title> <p>India is the world’s largest producer of sugar, with an annual production capacity of 29 million tonnes. Each crushing season, this intern produces over 10 million tonnes of pressmud, which is difficult to dispose of due to its inherent properties. The present study is part of larger investigation for treatment and disposal of pressmud and spentwash. Further, scope of this research article is confined to utilization of pressmud for aerobic composting of pressmud along with selected microbial consortium and stabilized spentwash. Composting was carried out in an open area with 50 kg of pressmud and 1% (w/w) dosage of microbial consortium. Stabilized spentwash was used at concentrations of 25, 50, 100, 150 and 200% (v/w) and applied at predetermined time intervals. The entire study lasted for 50 days and the results were compared to those recommended by the Fertilizer Control Order (FCO), Ministry of Agriculture, Government of India (1). In an organic compost, the FCO recommends a minimum concentration of 12%, 0.80%, 0.40%, and 0.40% in TOC, TKN, phosphorous and potassium, with a maximum C/N ratio of 20. During composting, the addition of 150% (CH5) stabilized spentwash resulted in a maximum nutrient concentration in the majority of the parameters analysed. CH5 showed that the concentration of TOC, TKN, C/N, phosphorous and potassium were 25.92±2.19%, 2.16±0.29%, 12.28±0.66, 6.55±0.11% and 15.90±1.37% respectively. Hence, it can be concluded that selected microbial consortium is capable of decomposing the organic matter found in pressmud. Additionally, the application of stabilized spentwash enhanced the nutritional content of end product.</p> </abstract>ARTICLE2022-01-24T00:00:00.000+00:00SCM-198 Can Regulate Autophagy Through the Bax/Bcl-2/TLR4 Pathway to Alleviate Renal Ischemia-Reperfusion Injury<abstract><title style='display:none'>Abstract</title><p>Renal ischemia-reperfusion (I/R) injury is frequently observed in several clinical cases. In this study, we want to investigate that SCM-198 attenuates renal injury in the renal I/R model and find out the possible mechanisms. Wistar albino 40 male rats were classified into four groups (n=10): control, DMSO, I/R, and SCM-198 30 mg/kg. In the group 4, SCM-198 was administered intraperitoneally once at the doses of 30 mg/kg following the reperfusion. Glomerular associated proteins (PCX), tubular damage factors (NGAL, KIM-1), blood urea nitrogen (BUN), serum creatinine, inflammatory cytokines (IL-1β, IL-18, and TNF-α), Bax/Bcl-2, TLR4, LC3B, and Beclin-1 were evaluated. SCM-198 played an essential role in mitigating kidney damage. SCM-198 alleviated tubular damage and decreased IL-1β, IL-18, and TNF-α levels. SCM-198 reduced the apoptosis marker Bax/Bcl-2 ratio, immune system protein TLR4, and autophagy proteins LC3B and Beclin-1. In brief, our results support the notion that SCM-198 has protective effects on I/R-induced renal injury. SCM-198 therapy may be a new alternative for the prevention and treatment of renal I/R injury.</p></abstract>ARTICLE2021-10-21T00:00:00.000+00:00Large scale propagation and in vitro weaning for the restoration of to support assisted colonisation of a threatened butterfly<abstract><title style='display:none'>Abstract</title><p>The distribution and abundance of <italic>Boloria selene</italic> (small pearl-bordered fritillary butterfly, SPBF) declined over recent decades in many parts of the UK. Availability of food plants, especially marsh violet (<italic>Viola palustris</italic>), for the caterpillars of the SPBF has been identified as one of the major causes of this decline. To achieve augmentation of existing colonies and develop new populations of SPBF large numbers of marsh violet propagules were required specifically to feed the larvae. The main objectives of the study were to produce thousands of good quality marsh violet propagules to restore selected habitats in the Heart of Durham, Northern England, using in vitro methods. Preliminary trials showed that in vitro multiplication of seedlings from wild collected seeds through conventional agar-based cultures was lengthy, expensive and turned out to be a non-viable route to achieve the objectives. This study explored the potential of bioreactor-based cloning and cost-effective one step rooting and weaning. Robust propagules, ready for transplantation following rapid propagation and one step rooting and weaning in vitro, were raised in a plug system for transplantation and establishment under field conditions. This was achieved by using simple and cost-effective methods to support the large-scale restoration exercise using 14,000 propagules. Application of high throughput micropropagation and low cost one step weaning systems for time-bound conservation and restoration projects are discussed in detail. This research highlights the important role of in vitro methods to support integrated biodiversity conservation of a native larval host plant and threatened butterfly.</p></abstract>ARTICLE2021-10-21T00:00:00.000+00:00Influence of scarification method on seed germination of the terrestrial orchid (Lam.)<abstract><title style='display:none'>Abstract</title><p>A critical step during <italic>in vitro</italic> sexual propagation of terrestrial orchids is the treatment of the microscopic seeds with a disinfecting solution that kills bacteria and fungi attached to the seeds. This treatment is necessary to prevent infection of the culture vessels. At the same time, the treatment serves to scarify the seeds, a process that disrupts seed dormancy and initiates germination. The literature is inconclusive with respect to the proper combination of disinfecting solution strength and treatment duration. Both factors should be adapted to each species to guarantee minimal infection rate without damaging the embryo. This research aims to compare three disinfection/scarification methods for seeds of <italic>Anacamptis laxiflora</italic> (Lam.): (i) soaking in 0.5% NaClO, (ii) soaking in 0.5% NaClO, then centrifugation, and (iii) presoaking the seeds in sucrose solution, then soaking in 0.5% NaClO. The seeds were soaked in the disinfecting solution for 5 to 85 min. Following scarification, the seeds were sown in modified Malmgren nutrient medium. Infected and germinated vessels were counted at 41 and 189 d after sowing. We found that the longer the chemical treatment, the lower the infection rate, and the higher the germination rate. There was no significant difference in germination rate between the NaClO and the NaClO-plus-centrifugation method; in fact, the slight savings in disinfection time effected by centrifugation were more than offset by the added complexity of the method. Moreover, we found that centrifugation significantly delays germination. The sucrose presoak-plus-NaClO method was superior to plain NaClO, as the sucrose stimulates the germination of microbial spores on the surface of the seeds, making them easier to kill. Perhaps seeds with thicker testa as well as whole immature capsules could benefit even more from the pretreatment in sucrose solution.</p></abstract>ARTICLE2021-01-22T00:00:00.000+00:00Investigation of the Therapeutic Effects of Chloroquine in Adriamycin-Induced Hepatotoxicity<abstract><title style='display:none'>Abstract</title><p>The aim of this study is to investigate the therapeutic effects of Chloroquine (CLQ) against Adriamycin (ADR) induced hepatotoxicity. ADR is a chemotherapeutic agent used in the treatment of many cancer types, but it causes hepatotoxicity. CLQ is used as an anti-inflammatory drug in the treatment of malaria, rheumatoid arthritis, and pneumonia caused by Covid-19. Rats were divided into four groups: Control group, ADR group (2 mg/kg Adriamycin, one in three days for 30 days, i.p.), CLQ group (50 mg/kg Chloroquine, per day for 30 days, i.p.), ADR+CLQ (2 mg/kg Adriamycin, one in three days for 30 days, i.p. and 50 mg/ kg Chloroquine, per day for 30 days, i.p.). Animals were sacrificed, and liver tissues were extracted for further examinations. Histopathological changes in liver tissues were scored and <italic>IL-17</italic> immunostaining was performed to determine the expression levels among experimental groups. Bodyweights in the ADR group decreased significantly compared to the Control group and CLQ group. Furthermore, bodyweight in ADR+CLQ group was significantly higher compared to ADR group. The histopathological score was significantly higher in ADR group when compared to Control and CLQ group while CLQ administrations reduced the damage induced by ADR in the ADR+CLQ group. <italic>IL-17</italic> immunoreactivity was considerably increased in the ADR group. On the other hand, <italic>IL-17</italic> expressions of ADR+CLQ were substantially less compared to ADR group. We suggest that CLQ can be used as a therapeutic agent to reduce the detrimental effects of ADR, thanks to its anti-inflammatory properties.</p></abstract>ARTICLE2021-01-22T00:00:00.000+00:00COVID-19 vaccines: Where do we stand?<abstract><title style='display:none'>Abstract</title><p>Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) was first reported in the city Wuhan, China in December 2019. The high rates of infection led to quick spread of the virus around the world and on March 11<sup>th</sup>, 2020, the World Health Organization (WHO) announced the pandemic of the Coronavirus disease 2019 (COVID-19) caused by the SARS-CoV-2. The pharmaceutical companies and institutions have been working towards developing a safe and effective vaccine in order to control the pandemic. The biology of SARS-CoV-2 is briefly discussed describing the transcription of the virus and the receptor recognition. The spike protein of SARS-CoV-2 is important in the attachment of the host cell and RNA-dependent RNA polymerase (RdRp) is involved in the replication of the virus making them good candidates for drug and vaccine targets. To date many different strategies have been employed in the development of vaccines and a number of them are in the phase III of clinical trials with promising results. In this mini-review, we assessed the literature throughly and described the latest developments in SARS-CoV-2 vaccines for humans. The main benefits and drawbacks of each platform is evaluated and the possible changes in the vaccine effectivity due to naturally occuring SARS-CoV-2 mutations have been described.</p></abstract>ARTICLE2021-01-22T00:00:00.000+00:00Recent Advances in 3D Printing of Polyhydroxyalkanoates: A Review<abstract><title style='display:none'>Abstract</title><p>In the 21<sup>st</sup> century, additive manufacturing technologies have gained in popularity mainly due to benefits such as rapid prototyping, faster small production runs, flexibility and space for innovations, non-complexity of the process and broad affordability. In order to meet diverse requirements that 3D models have to meet, it is necessary to develop new 3D printing technologies as well as processed materials. This review is focused on 3D printing technologies applicable for polyhydroxyalkanoates (PHAs). PHAs are thermoplastics regarded as a green alternative to petrochemical polymers. The 3D printing technologies presented as available for PHAs are selective laser sintering and fused deposition modeling. Stereolithography can also be applied provided that the molecular weight and functional end groups of the PHA are adjusted for photopolymerization. The chemical and physical properties primarily influence the processing of PHAs by 3D printing technologies. The intensive research for the fabrication of 3D objects based on PHA has been applied to fulfil criteria of rapid and customized prototyping mainly in the medical area.</p></abstract>ARTICLE2021-01-22T00:00:00.000+00:00Comparison of deep learning and conventional machine learning methods for classification of colon polyp types<abstract><title style='display:none'>Abstract</title><p>Determination of polyp types requires tissue biopsy during colonoscopy and then histopathological examination of the microscopic images which tremendously time-consuming and costly. The first aim of this study was to design a computer-aided diagnosis system to classify polyp types using colonoscopy images (optical biopsy) without the need for tissue biopsy. For this purpose, two different approaches were designed based on conventional machine learning (ML) and deep learning. Firstly, classification was performed using random forest approach by means of the features obtained from the histogram of gradients descriptor. Secondly, simple convolutional neural networks (CNN) based architecture was built to train with the colonoscopy images containing colon polyps. The performances of these approaches on two (adenoma &amp; serrated vs. hyperplastic) or three (adenoma vs. hyperplastic vs. serrated) category classifications were investigated. Furthermore, the effect of imaging modality on the classification was also examined using white-light and narrow band imaging systems. The performance of these approaches was compared with the results obtained by 3 novice and 4 expert doctors. Two-category classification results showed that conventional ML approach achieved significantly better than the simple CNN based approach did in both narrow band and white-light imaging modalities. The accuracy reached almost 95% for white-light imaging. This performance surpassed the correct classification rate of all 7 doctors. Additionally, the second task (three-category) results indicated that the simple CNN architecture outperformed both conventional ML based approaches and the doctors. This study shows the feasibility of using conventional machine learning or deep learning based approaches in automatic classification of colon types on colonoscopy images.</p></abstract>ARTICLE2021-01-22T00:00:00.000+00:00Are new genome variants detected in SARS-CoV-2 expected considering population dynamics in viruses?<abstract><title style='display:none'>Abstract</title><p>The pandemic COVID-19 is caused by a highly transmissible severe acute respiratory coronavirus 2 (SARS-CoV-2) which showed the highest morbidity and mortality rates among the other coronavirus infections such as SARS-CoV and MERS-CoV. However, the numbers of infected cases as well as mortality rates are varying from population to population. Therefore, scientist has urged the SARS-CoV-2 genome and host genetic factors investigations. Recently, new SARS-CoV-2 variants has been detected and though to affect the diseases transmission from human to human. In this mini-review, we aimed to explained detected SARS-CoV-2 variants that thought to influence the COVID-19 severity and transmission using the literature.</p></abstract>ARTICLE2021-01-22T00:00:00.000+00:00Vitamin D3 as possible diagnostic marker of Eating Disorders<abstract><title style='display:none'>Abstract</title><sec id="j_ebtj-2021-0005_s_006_w2aab3b7c20b1b6b1aab1c15b1Aa"><title style='display:none'>Purpose</title><p>Eating Disorders (EDs) refer to a group of psychiatric conditions in which disorderly food intake results in impaired psychological functioning or physical health. Nowadays, these disorders represent an increasing problem in modern society. There are no universally validated clinical parameters to confirm, disprove or simply help to identify EDs except for diagnostic criteria on psychiatric basis. The aim of this study was the assessment of Vitamin D3 level in patients with EDs to understand if it might be a valid clinical biochemistry parameter useful as prognostic marker.</p></sec><sec id="j_ebtj-2021-0005_s_007_w2aab3b7c20b1b6b1aab1c15b2Aa"><title style='display:none'>Methods</title><p>The sample consists of 28 female patients, who suffer from EDs. Blood samples were examined in terms of blood count, glucose, cholesterol and Vitamin D3 levels. The other clinical biochemistry parameters were analysed to understand if the Vitamin D3 was the only altered parameter.</p></sec><sec id="j_ebtj-2021-0005_s_008_w2aab3b7c20b1b6b1aab1c15b3Aa"><title style='display:none'>Results</title><p>The parameters that appear altered are glycemia, cholesterol and, in particular, Vitamin D3. Significant results were obtained comparing controls with restrictive-type anorexia nervosa (p value= 0,003) and with purging-type anorexia nervosa (p value= 0,007).</p></sec><sec id="j_ebtj-2021-0005_s_009_w2aab3b7c20b1b6b1aab1c15b4Aa"><title style='display:none'>Conclusion</title><p>There are currently no universally validated and diagnostic reliable clinical biochemistry parameters for EDs but, in the light of the findings, but our research indicates the potential use of Vitamin D3 as a biomarker for anorexia nervosa.</p></sec><sec id="j_ebtj-2021-0005_s_010_w2aab3b7c20b1b6b1aab1c15b5Aa"><title style='display:none'>Level of evidence</title><p>Level III: Evidence obtained from a single-center cohort study.</p></sec></abstract>ARTICLE2021-01-22T00:00:00.000+00:00Nile Red assay development for the estimation of neutral lipids in and<abstract><title style='display:none'>Abstract</title><p>Fluorescent dyes offer a useful method for the measurement of intracellular lipids. They are inexpensive and require simple optical measurement instrumentation, whilst simultaneously providing high throughput application. Nile Red is a hydrophobic, metachromatic dye which has been widely used for detection of intracellular lipids. However, Nile Red fluorescence depends on its concentration, microenvironment polarity, incubation time and, therefore, requires strain specific optimization. Hence, neutral lipids in <italic>Chlorella emersonii</italic> and <italic>Pseudokirchneriella subcapitata</italic> cannot be quantified using existing Nile Red methods developed for other microalgae strains and, therefore an optimised procedure for these strains is required. In this method development, the optimal excitation and emission wavelengths were selected based on the solvent used for Nile Red dissolution. The effect of Nile Red concentration, microalgae cell concentration, incubation time on fluorescence intensity was explored and optimised. Quintuplet assay repeats were executed for increased assay robustness for two microalgae strains, <italic>Chlorella emersonii</italic> and <italic>Pseudokirchneriella subcapitata</italic>, with protocol reliability confirmed by fluorescence microscopy. In brief, 20% (v/v) DMSO containing 10μg/ml and 5μg/ml Nile red was found to be ideal concentration for neutral lipid estimation in <italic>Chlorella emersonii</italic> and <italic>Pseudokirchneriella subcapitata</italic> respectively when an incubation time of 60mins and 40mins at 40°C was used. This optimised Nile Red protocol is a robust, simple and cost-effective method for neutral lipid quantification in <italic>Chlorella emersonii</italic> and <italic>Pseudokirchneriella subcapitata</italic>.</p></abstract>ARTICLE2020-10-21T00:00:00.000+00:00Revealing the therapeutic effects of aminolevulinate mediated femtosecond laser induced photo-chemotherapy in different cancer cells<abstract><title style='display:none'>Abstract</title><p>Photodynamic therapy (PDT) is a photo chemotherapeutic strategy that is the application of photosensitizing agent and light on disease or tumor site. The aim of this study is to confirm the feasibility for femtosecond (fs) laser for aminolevulinate (ALA) mediated PDT on skin, breast and bladder cancer cells. Also the remarkable aspects of ALA mediated and laser induced PDT with respect to other literally known applications were investigated.</p><p>Metastatic melanoma cells SK-MEL30, mammary epithelial carcinoma cells MCF-7 and bladder cancer cells UMUC-3 were treated with ALA and then the cells were irradiated by fs laser at thirty wavelengths in between 230 and 800 nm for 30s and 60s. Anti-cancer effects of ALA phototherapy on different cancer cell lines were determined. Protoporphyrin IX (PpIX) accumulation was visualized by confocal microscopy. The effective PDT wavelengths were applied to evaluate the degree of apoptosis and necrosis in cells.</p><p>The viability tests demonstrated that wavelengths 400-440 nm and 600-630 nm were found to decrease the viability on three model cell lines. PDT at 630 nm exerted cell death by necrosis and apoptosis after 30 s and 60 s periods.</p><p>This paper confirms that ALA and femtosecond laser mediated PDT may be used together as therapeutic and diagnostic method to target breast, skin and urinary bladder cancer cells. The use of fs laser allows the flexibility for optimization of wavelength for photosensitizing agents.</p></abstract>ARTICLE2020-10-21T00:00:00.000+00:00en-us-1