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It is imperative to emphasize the wear-reducing and load-sustaining abilities of the lubricating oils, as they are an essential part of their operation. In this study, the anti-wear and extreme pressure (EP) performance of Jatropha oil was explored. The investigation was executed on a 4-ball tester machine according to ASTM-D-4172 and D-2783 standards. Nano-sized Molybdenum disulfide and graphene were added to the base oil, whose addition reduced the wear on the test balls but failed to improve the EP properties. However, the addition of Garlic oil elevated the EP properties of the base oil. The Last Non-seizure load and weld point of the jatropha oil was elevated by up to 3 levels, while the Load Wear index was increased by 91.88%. The Flash Temperature Parameter of jatropha oil was increased by 658% and 250% at 100kgf and 126kgf load, respectively, after the addition of 5% garlic oil. Further, the wear scars were examined under an optical microscope and FESEM, which proved that the addition of nano-additives reduced the wear of the balls during anti-wear tests at lower loads, and garlic oil reduced the wear in EP conditions and produced smoother surfaces. These improvements were attributed to the formation of tribo-films on the surfaces as characterized by the EDS analysis. It was concluded that Garlic oil could be a substantial EP additive to vegetable oils (especially jatropha oil) to lift their load-carrying capacity without hampering their environmentally friendly nature.

Due to solar PV panels' growing feasibility as a renewable energy source, PV panel installations have surged recently. Machine learning algorithms can now generate better predictions due to the increasing availability of data and computing capacity. For many stakeholders in the energy business, predicting solar PV energy output is crucial, therefore machine learning and time series models may be used to do this. In this study, time series models and several machine learning techniques are compared across five different sites in India. Since the energy time series are non-stationary, we find that applying time series models is a challenging process. On the other hand, putting machine learning methods into practice was simpler.

Better prediction designs for the contributes directly of renewables are essential to reducing the requirement for controlling the energy associated with traditional power plants. For the extremely variable wind power output to be successfully integrated into the power system, a reliable forecast is especially crucial. For a wind inclusion study in the western United States, we use data from the National Renewable Energy Laboratory (NREL) and focus on summary wind power projection in this chapter. Our approach immediately derives functional connections from data, in contrast to physical procedures that depend on exceedingly complex differential equations. By reprising the regression model as a regression issue, we explore several regression strategies including regression models, k-nearest peers, and support vector regression. In our experiments, we look at forecasts for both individual wind turbines and entire wind parks, proving the viability of a machine learning technique for short-term wind power projection

Lumpy skin disease (LSD) is a highly infectious disease of cattle caused by a virus of the Poxviridae family, genus Capripoxvirus. The present study was designed to determine the prognostic ability of serum IL-6 in LSD using a binary logistic regression model at baseline sampling. A 17-day cohort study was conducted on a recent outbreak of LSD among cattle in the Lodhran District of Punjab, Pakistan. Infected cattle were divided into two categories based on their clinical status on day 17 as recovered (n = 33) or unrecovered (n = 17). Nodular lesions and scab specimens (n = 50) were used for the isolation of the lumpy skin disease virus and were confirmed by PCR. In recovered animals, hematological results showed marked leukocytosis, eosinophilia, lymphocytosis, neutrophilia, and monocytopenia. However, marked erythrocytosis, leukopenia, and thrombocytopenia were observed in the unrecovered animals at the final sampling point of the study. Serum levels of total protein, albumin, and glucose were significantly higher in the recovered animals. Meanwhile, aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase, creatinine phosphokinase, total bilirubin, and direct bilirubin were found considerably higher in the unrecovered group. Receiver-operating characteristic curve analysis for serum IL-6 at baseline predicts the extended clinical conditions at the cut-off value of 85.16 pg/mL (55% specificity, 94% sensitivity, area under the curve 0.8039, respectively). In conclusion, the disease-induced hematological and biochemical alterations were significantly ameliorated in the recovered animals. In addition, the study revealed that serum IL-6 can be used as a valid marker for predicting the clinical worsening of LSD in cattle.

Melia azedarach is a medicinal plant utilized for the treatment of various disorders in Asian countries. The antioxidant potential of fruit-seeds from this plant was investigated using n-hexane, chloroform, methanol and aqueous extracts. Preliminary studies were performed on powder as well as on various extracts. An in vitro anti-oxidant activity was evaluated using DPPH, ABTS and TAC assays. All aforementioned extracts were rich in flavonoids and polyphenols. The aqueous extract (61.30 ± 3.55%) showed high DPPH scavenging activity followed by methanol (52.95% ± 0.53%), chloroform (52.61 ± 3.59%), and n-hexane extract (51.16 ± 1.70%). Similarly, all the extracts exhibited high ABTS scavenging activity. However, in TAC assay, chloroform extract (81.16 ± 1.09%) exhibited the highest inhibitory activity, followed by aqueous, methanol and n-hexane. Therefore, further isolation and characterization of the plant is required to explore the bioactive compounds for their anti-oxidant activity

Background Bifenthrin is an insecticide and anti-estrogenic compound primarily used to control residential pests by depolarizing sodium gated voltage channels in the nervous system. Eryptosis, the suicidal death of erythrocytes, featured by PS exposure, membrane blebbing and cell shrinkage. Anemia is an outcome of uncontrolled eryptosis. Research Design In this study, erythrocytes were treated with different concentrations (.5-1-1.5 μM) of bifenthrin over a period of 48 hours. In order to investigate the oxidative stress induced by bifenthrin, catalase, superoxide dismutase, and glutathione peroxidase activities were investigated. Results Obtained data indicated the decrease in the enzymes (superoxide dismutase, glutathione peroxidase, and catalase) activities in bifenthrin treated cells at 1 μM concentration. In addition, measurement of cell size and confirmation of the role of calcium in the stimulation of the eryptotic activity of bifenthrin were performed. A significant increase in mean cell volume was found in the presence of bifenthrin and a decrease in mean cell volume in the presence of calcium channel blocker was observed. Similarly, there was also a significant increase in the percentage of hemolysis indicating the necrotic activity of bifenthrin. Conclusions It is concluded that the indicated doses of bifenthrin triggered oxidative stress which may lead to early cell death by eryptosis and hemolysis.

Biodiesel has attracted considerable interest as an alternative biofuel due to its many advantages over conventional petroleum diesel such as inherent lubricity, low toxicity, renewable raw materials, biodegradability, superior flash point, and low carbon footprint. However, high production costs, poor low temperature operability, variability of fuel quality from different feedstocks, and low storage stability negatively impact more widespread adoption. In order to reduce production costs, inexpensive inedible oilseed alternatives are needed for biodiesel production. This study utilized inedible tamarind (Tamarind indica) seed oil as an alternative biodiesel feedstock, which contained linoleic (31.8%), oleic (17.1%), and lauric (12.0%) acids as the primary fatty acids. A simple and cost-effective high vacuum fractional distillation (HVFD) methodology was used to separate the oil into three fractions (F1, F2, and F3). Subsequent transesterification utilizing basic, acidic, and enzymatic catalysis produced biodiesel of consistent quality and overcame the problem of low temperature biodiesel performance. The most desirable biodiesel with regard to low temperature operability was produced from fractions F2 and F3, which were enriched in unsaturated fatty acids relative to tamarind seed oil. Other properties such as density and cetane number were within the limits specified in the American and European biodiesel standards.

Nano-materials offer potential scope for an increasing numerous novel applications when engineered to deliver availably functional properties. The nanosized additives when added to biodegradable oils improve their tribological performance and contribute to energy saving and sustainability. In the present study, the MoS2 nanoparticles with different mass ratios were employed as lubricant additives in the base jatropha oil, and their tribological properties were evaluated using a reciprocating ball-on-disc tribometer for steel-steel contacts. The results demonstrate that the MoS2 nanoparticles exhibit superior lubrication performance. The optimal concentration of MoS2 nanoparticles in the base oil was found to be 0.5% for minimum friction and wear rate. Addition of load decreased friction and increased wear rate. The coefficient of friction and wear rate was reduced by 63% and 35%, respectively. The excellent lubrication properties of the MoS2 nanoparticles are attributed to the physical synergistic lubricating actions of nano-MoS2 during the rubbing process.

Fibrinolytic therapy progressed by the evolution of different strategies that helped in enhancing its efficacy and specificity. The use of microbial fibrinolytic enzymes is leading to a promising direction for the treatment of cardiovascular diseases. Subtilisin, a member of subtilases is a fibrinolytic enzyme abundantly found in Bacillus species. The isolation of subtilisin gene (quaking homolog) from Bacillus subtilis was attempted in the present work. The genomic deoxyribonucleic acid extraction was done following Yamada protocol and used as a template for polymerase chain reaction amplification of the target gene using specifically designed primers. The polymerase chain reaction product was ligated into cloning vector pTZ57R/T followed by its transformation into Escherichia coli top 10 strain. A 1090 base pair, partial gene sequence was amplified coding for subtilisin protein of 363 amino acids with molecular weight of 37550.7 Daltons. The nucleotide sequence revealed significant evolutionary relationships with subtilases from other strains of Bacillus subtilis. Our study confirms the presence of subtilisin (quaking homolog) gene in local Bacillus species suggesting economical way to produce important thrombolytic agents of commercial and pharmaceutical worth.

The emergence of a pathogen responsible for a mysterious respiratory disease was identified in China and later called a novel coronavirus. This disease was named COVID-19. The present study seeks to determine the epidemiological and clinical characteristics of COVID-19 in Pakistan. This report will exhibit a linkage between epidemiology and clinical aspects which in turn can be helpful to prevent the transmission of the virus in Pakistan. A retrospective, multiple center study was performed by collecting the data from patients' with their demographics, epidemiological status, history of co-morbid conditions, and clinical manifestations of the disease. The data was collected from 31 public-sector and 2 private hospitals across Pakistan by on-field healthcare workers. A Chi-square test was applied to assess the relationship between categorical data entries. A total of 194 medical records were examined. The median age of these patients was found to be 34 years. A total of 53.6% active cases were present including 41.2% males and 12.4% females till the end of the study. Adults accounted for most of the cases (94.3%) of COVID-19. Fever (86.60%), cough (85.05%), fatigue (36.60%), dyspnea (24.74%), and gastrointestinal discomfort (10.31%) were among the most frequently reported signs and symptoms by the patients. However, 4.12% of the total patient population remained asymptomatic. The median duration of hospital stay was found to be 14 (0–19) days. The earliest source of the spread of the virus may be linked to the foreigners traveling to Pakistan. Spread among men was more as compared to women. A few cases were found to be positive, due to the direct contact with pets or livestock. Hypertension (7.73%), diabetes (4.64%), cardiovascular conditions (2.58%) were the most common co-morbidities. The percentage mortality was 2.50% with the highest mortality among elders.

Crude oil reserves are sapping with time, therefore researchers have considerably shifted their focus to bio-lubricants which can be surrogated for petroleum-based lubricants. The objective of the present research is to cater an idea to the researchers about the lubricating potentials of Jatropha oil (JO). The study was performed using reciprocating friction monitor (RFM) machine having ball-on-flat arrangement with 52100 chromium steel ball and EN-31 steel as a tribo-pair. The experiments were conducted at three different loads at a temperature of 40 °C. JO was mixed with glycerol (G) and different percentages of molybdenum disulphide (0.5%, 1%, 2%, and 3%) micro-particles to prepare a lubricant. The friction and wear of the tribo-pair was assessed in the presence of these mixtures. The results revealed that MoS2 micro-particles were effective in reducing both friction and wear, with minimum friction recorded at 2% MoS2 concentration. The coefficient of friction (COF) was reduced by 18–52% with the addition of MoS2 as compared to the base JO. The results showed that JO has better lubrication ability as compared to the commercial lubricant SAE 10W/30. The experimental results were substantiated by examinations of tribo-pairs on scanning electron microscope, optical microscope, and optical three-dimensional profilometer. The results were found to be in close agreement with each other. It was concluded that the blend (80% JO + 20% G + 2% MoS2) is a strong candidate that can be used in the place of mineral oils as a lubricant.

Omeprazole, a proton pump inhibitor blocks the H+/K+-ATPase channels of gastric parietal cells. It is used for the treatment of peptic ulcer. Prolonged use of omeprazole may involve in inducing anemia. The key marker of eryptosis includes membrane blebbing, cell shrinkage and phosphatidylserine (PS) exposure at the cell surface. In current study, the eryptotic, oxidative as well as hemolytic effects of therapeutical doses (0.5, 1 and 1.5 µM) of omeprazole were investigated after exposing erythrocytes for 48 hours. Investigation of eryptosis was done by cell size measurement, PS exposure determination and calcium channel inhibition. As a possible mechanism of omeprazole induced eryptosis, oxidative stress was investigated by determining the catalase, glutathione peroxidase and superoxide dismutase activities. Similarly, necrotic effect of omeprazole on erythrocytes was also evaluated through hemolysis measurement. Results of our study illustrated that 1.5 µM of omeprazole may induce significant decrease in superoxide dismutase, glutathione peroxidase and catalase activities as well as triggered the erythrocytes shrinkage, PS exposure and hemolysis. Role of calcium was also confirmed in inducing erythrocyte shrinkage. It is concluded that the exposure of erythrocytes with 1.5 µM omeprazole may enhance the rate of eryptosis and hemolysis by inducing oxidative stress.

Biodiesel offers an advantage only if it can be used as a direct replacement for ordinary diesel. There are many reasons to promote biodiesel. However, biodiesel cannot get wide acceptance until its drawbacks have been overcome including poor low temperature flow properties, variation in the quality of biodiesel produced from different feedstocks and fuel filter blocking. In the present study, a much cheaper and simpler method called high vacuum fractional distillation (HVFD) has been used as an alternative to produce high-quality refined biodiesel and to improve on the abovementioned drawbacks of biodiesel. The results of the present study showed that none of biodiesel sample produced from crude Azadirachta indica (neem) oil met standard biodiesel cetane number requirements. The high vacuum fractional distillation (HVFD) process improved the cetane number of produced biodiesels which ranged from 44–87.3. Similarly, biodiesel produced from fractionated Azadirachta indica oil has shown lower iodine values (91.2) and much better cloud (−2.6 °C) and pour point (−4.9 °C) than pure Azadirachta indica oil. In conclusion, the crude oil needs to be vacuum fractioned for superior biodiesel production for direct utilization in engine and consistent quality production.

Humans are experiencing serious health issues like myocardial infarction and diabetes. Thrombosis is the reason of myocardial infarction that may cause death. Bioactive compounds or enzymes can be used to dissolve the clot. Whereas diabetes is a disorder of metabolism in which the level of glucose in blood becomes high. It can be controlled by inhibiting α-amylase enzyme. The current project was, therefore planned to investigate the thrombolytic, α-amylase inhibitory and cytotoxic (to access drug safety) potentials of the organic and aqueous bioactive fractions of Bacillus clausii KP10. The cytotoxicity was assessed with hemolytic assay, α-amylase inhibition assay was done by using DNS and in-vitro thrombolytic effect was checked with human blood. In our experiments, the maximum hemolytic activity was shown by ethyl acetate fraction (12.64%). Results were compared with standard Triton X-100 which showed 91.61% hemolytic activity whereas all other fractions showed least cytotoxic activity. The extracts were also evaluated as thrombolytic agents as correlated to streptokinase (73.83%). All the extracts showed clot lysis activity, among which water soluble fraction exhibited maximum (35.16%) clot lysis activity. In our experiment methanol soluble fraction of B. clausii KP10 showed maximum 26.49% α-amylase inhibitory activity. Results were analyzed statistically through analysis of variance (ANOVA).

The aim of the research work was to synthesize different biologically active carbamate derivatives bearing 2-furoyl-1-piperazine and having modest toxicity. The synthesis was completed in multiple steps. The structural confirmation of all the synthesized compounds was carried out by EI-MS, IR and 1 H-NMR spectral data. The enzyme inhibition and antibacterial potential of the synthesized compounds was evaluated. Moreover, the cytotoxicity of the compounds was also checked to find their utility as possible therapeutic agents. All the compounds were active against acetylcholinesterase enzyme especially 12 and 14 showed very good inhibitory potential relative to Eserine, a reference standard. Almost all the compounds showed good activities against the both Gram-positive and Gram-negative bacterial strains.

Novel therapeutic agents discovery have become crucial because humankind is experiencing issues regarding health and the environment. The present study is focusing on thrombolytic agents, which are therapeutics for cardiovascular diseases, eco-friendly, safe and can be used in comparison to synthetic agents that are associated with some side effects and their production is not environment friendly. The main objective of the present research was to evaluate the thrombolytic and cytotoxic potential of different extracts of Escherchia coli-ATCC35218. Soxhlet extraction method with ethanol was used for extraction from cells. Crude protein was extracted from the cell mass with protein extraction buffer. Crude isoprenoid quinone and polar lipids were extracted by using petroleum ether with methanol and chloroform as extraction solvent. In another method, the extracts were prepared using methanol as well as n-hexane, chloroform, ethyl acetate and water in serial extraction. These organic and aqueous extracts of E. coli were dried, weighed, dissolved in DMSO and tested for their hemolytic and thrombolytic potential. Mild to moderate percentage of thrombolytic activity was shown by these extracts and compared with citric acid (standard). Chloroform, ethyl acetate and methanol soluble extracts showed significant thrombolytic activities that were 43.5% ​± ​0.06, 41.8% ​± ​0.16 and 40.5% ​± ​0.07 respectively, while citric acid showed 73.7% ​± ​0.05. Maximum hemolytic activities were shown by polar lipids, crude methanol extract and n-hexane that are 87.20% ​± ​0.08, 82.5% ​± ​0.06 and 73.46%± ​0.08 respectively as compared with 0.1% Triton X-100 (positive control) with 94.75% ​± ​0.15 hemolysis. Finally, results were analyzed statistically through analysis of variance (ANOVA). Further studies on isolation and identification of bioactive thrombolytic agents from each bioactive thrombolytic extract will help us in reporting natural thrombolytic agents from E.coli that could benefit human medically. This is one of the few reports of thrombolytics available from natural sources.

Cardiac patients with weak immune system are susceptible to bacterial infections. Their prescriptions frequently contain simvastatin and clarithromycin together. The objective of present project was to assess the potential interaction between simvastatin and clarithromycin by evaluating the clarithromycin effects on the pharmacokinetics of simvastatin in healthy adult male subjects. The study design comprised of two phases, used at interval of one week. In first phase simvastatin 20 mg alone was administered to each volunteer. In second phase, co-administration of simvastatin 20 mg with clarithromycin 250 mg was made under similar specified conditions. Blood samples were collected at specified time intervals. Simvastatin plasma concentrations were analyzed through High Performance Liquid Chromatography with UV detector at 238 nm wavelength. Using one compartment open model, MW/PHARM version 3.02 software program was used by F. Rombut for pharmacokinetic parameters calculation. Clarithromycin co-treatment resulted in 2.3 fold increase in maximum plasma concentration Cmax (from 2.47±0.34 ng.mL

In aromatic plants species, biosynthesis of essential oils occurs through two complex natural biochemical pathways involving different enzymatic reactions. Isopentenyl diphosphate (IPP) and its isomer dimethylallyl diphosphate (DMAPP) are the universal precursors of essential oil biosynthesis and are produced by the cytosolic enzymatic MVA (mevalonic acid) pathway or by plastidic and enzymatic 1-deoxy-d-xylolose-5-phosphate (DXP) pathway, also called the 2-C-methylerythritol-4-phosphate (MEP) pathway. In the particular plant cell part, prenyl diphosphate synthases condense isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) further to form prenyl diphosphates, which are used as substrates for geranyl diphosphate (GPP; C10) or for fernesyl diphosphate (FPP; C15). Essential oils are final terpenoid products and are formed by a huge group of enzymes known as terpene synthases (TPS). Essential oils are important secondary metabolites of plants and have been used not only in different i...

Terpenoids are synthesized in plants through the mevalonate (MVA) and the methylerythritol phosphate (MEP) pathways, among isopentenyl diphosphate (IPP) as the central intermediate metabolite. 3-Hydroxy-3-methylglutaryl- Coenzyme A synthase (HMGS) is a subsequent enzyme in MVA pathway of isoprenoid biosynthesis and catalyses the condensation of acetyl-CoA by acetoacetyl-CoA to yield HMG-CoA. Different genes of HMGS are present in different species of plants and animals for the synthesis of secondary metabolites. The present study was planned to compare and analyse bioinformatically the HMG-CoA gene sequence of Brassica rapa with other sequences of different species. The gene sequence have 925 bp coding DNA sequence and was submitted in National Centre for Biotechnology Information (NCBI) with accession Number. MF577077. The alignment of nucleotide and protein sequences was carried out through Geneious. Bioinformatics comparison including structural and functional analysis of HMG-CoA synthase like gene sequence isolated from Brassica rapa was carried out by using different bioinformatics tools and softwares. These comparisons gave a better information about properties, genetic and functional diversity of this gene along with amino acid substitutions.

Bacillus strains have been widely used for the production of fibrinolytic enzymes having role in the treatment of cardiovascular disorders. Purification and overproduction of such enzymes has increased their usage in medical fields including metalloproteinases with the ability to degrade extracellular matrix (ECM). Camelysin, a neutral metalloproteinase has been isolated from different species of bacteria like Bacillus cereus, Bacillus anthracis, and Bacillus thuringiensis with fibrinolytic, collagenolytic and actin degradation activity. This project successfully demonstrated the presence of 734-bp coding DNA sequence (CDS) encoding a 20.72331 kDa camelysin gene in local strain of Bacillus thuringiensis containing a signal peptide with cleavage site between residues 19 and 20. The sequence was submitted to GenBank (KT023597) and the sequence showed high homology with the camelysin protein of closely related Bacillus species. The alignment of related proteins through ClustalW displayed difference of four amino acids (“Q” replaced by “P” at position 169 and at position 182–184, “NQE” replaced by “HLK”) in the isolated protein. Comparison including structural and functional analysis of camelysin sequences isolated from different Bacillus species was carried out using different bioinformatics tools and software. The information would help in better understanding the properties of camelysin protein and its role in pathogenicity and clinical treatments.

The expression of xylanases and cellulases is under carbon catabolite repression, a regulatory repression mechanism of transcription of these enzymes caused bycre1 gene. In the present study we isolatedcre1 partial gene from a thermophilic fungusChaetomium thermophilum ATCC 28076. The fungus was grown on Eggins and Pugh medium with glucose as a carbon source. Genomic DNA was isolated by two different methods and the integral DNA was subjected to polymerase chain reaction (PCR) for the amplification ofcre1 partial gene sequence. The PCR product was ligated into pTZ57R/T vector and transformed in E.coli TOP 10 strain. cDNA from total RNA was utilized as template for RT-PCR analysis, which confirmed the presence of an intronless cre1 gene in the model organism. This is the first report on isolation ofcre1 gene fragment in C.thermophilum.

Essential oils are industrially important natural products of aromatic plants, which are biosynthesized in specialized cells types, such as osmophores, Glandular Trichomes, and ducts and cavities, present on different parts of these plants. In these cells, the essential oils are biosynthesized, accumulated and transferred to the atmosphere by different reported secretion mechanisms. Numerous environmental factors also affect biosynthesis of essential oils and their secretion in the atmosphere. A deep understanding of the entire process is important to improve the yield of biomass of aromatic plants and thus produce large quantities of commercial volatile oils. In this paper, the recent research on types and structure of essential oil-bearing cells, secretion mechanisms, and effect of different environmental factors on essential oil biosynthesis have been comprehensively reviewed. Therefore, this review article will provide new research directions for the researchers working in the field of aromatic plants physiology, essential oil production and many others applications.

In the present work, a new series of different 4-{[4-(2-furoyl)-1-piperazinyl]methyl}-N-(substituted-phenyl) benzamides (5a-h) have been synthesized as possible therapeutic agents for the treatment of Alzheimer’s disease. The structural confirmation of all the synthesized compounds was carried out by their IR, 1H-NMR and EI-MS spectral data. Enzyme inhibition activity was performed against butyrylcholinestrase enzyme, which revealed that, 4-{[4-(2-furoyl)-1- piperazinyl]methyl}-N-(4-ethylphenyl)benzamide (5b) showed excellent IC50 value 0.82 ± 0.001 μM relative to Eserine, a reference standard having IC50 value of 0.85 ± 0.0001 μM. The enhanced potential of this molecule may be attributed to the 4-ethylphenyl group. As the cholinesterase enzyme inhibitors are good targets for Alzheimer’s disease, therefore, the inhibition study of these synthesized molecules was carried out to discover their possible therapeutic effect as target for aforesaid disease.

We report for the first time the recombinant expression of fully folded bioactive cyclotides inside live yeast cells by using intracellular protein trans-splicing in combination with a highly efficient split-intein. This approach was successfully used to produce the naturally occurring cyclotide MCoTI-I and the engineered bioactive cyclotide MCoCP4. Cyclotide MCoCP4 was shown to reduce the toxicity of human α-synuclein in live yeast cells. Cyclotide MCoCP4 was selected by phenotypic screening from cells transformed with a mixture of plasmids encoding MCoCP4 and inactive cyclotide MCoTI-I in a ratio of 1:5×10(4). This demonstrates the potential for using yeast to perform phenotypic screening of genetically encoded cyclotide-based libraries in eukaryotic cells.