A lot of kitchen waste oil is produced every day worldwide, leading to serious environmental pollution. As one of the environmental protection methods, microorganisms are widely used treating of various wastes. Lipase, as one of the cleaning agents can effectively degrade kitchen waste oil. The composting process of pig carcasses produces many lipase producing microorganisms, rendering compost products an excellent source for isolating lipase producing microorganisms. To our knowledge, there are no reports isolating of lipase producing strains from the high temperature phase of pig carcass compost.Lipase producing strains were isolated using a triglyceride medium and identified by 16S rRNA gene sequencing. The optimal fermentation conditions for maximum lipase yield were gradually optimized by single-factor tests. The extracellular lipase was purified by ammonium sulfate precipitation and Sephadex G-75 gel isolation chromatography. Amino acid sequence analysis, structure prediction, and molecular docking of the purified protein were performed. The pure lipase's enzymatic properties and application potential were evaluated by characterizing its biochemical properties.Bacillus sp. ZF2 was isolated from pig carcass compost products, the optimal fermentation conditions of lipase: sucrose 3 g/L, ammonium sulfate 7 g/L, Mn2+ 1.0 mmol/L, initial pH 6, inoculum 5%, temperature 25 ℃, and fermentation time 48 h. After purification, the specific activity of the purified lipase reached 317.59 U/mg, a 9.78-fold improvement. Lipase had the highest similarity to the GH family 46 chitosanase and molecular docking showed that lipase binds to fat via two hydrogen bonds at Gln146 (A) and Glu203 (A). Under different conditions , lipase can maintain enzymatic activity. Under different types of kitchen oils, lipase has low activity only for ‘chicken oil’, in treating other substrates, the enzyme activity can exceed 50%.In this study, a lipase producing strain of This study reveals the potential of lipase for waste oil removal, and future research will be devoted to the application of lipase.The online version contains supplementary material available at 10.1186/s12896-023-00818-1. Pseudomonas brenneri, and the produced lipase can be used to synthesize fatty acid methyl esters [Fusarium incarnatum KU377454, and after optimization, lipase activity could reach 38.35 U/ml. A rod-shaped Bacillus amylolyticus was isolated from soil, and the lipase production could reach 80.63 U/ml when incubated at 40 ℃, pH 7.0, and 3–5% inoculum for 72 h. Mandari et al. isolated Aspergillus niger MTCC 872 with 269.87 ± 8.09 U/ml lipase activity at 35 ℃, pH 7, and 70% initial water content [Around the world, large quantities of used cooking oil are generated every day , used col esters . Ritika  content . The com content . In addi content . The hig content . To our Bacillus sp.. It was purified by optimizing its enzyme production conditions. Next, the purified lipase was subjected to identification, structure prediction, and molecular docking as well as a series of biochemical characterization experiments, and the enzyme properties, thermal stability, and enzyme activity of the lipase were analyzed in the treatment of different animal and plant fats and oils.In this study, a lipase producing bacterium was isolated from the high temperature composting stage of pig carcasses and identified as 3COONa-3H2O 1.0 g/L, NH4Cl 1.0 g/L, NaCl 1.0 g/L, KH2PO4 0.5 g/L, K2HPO4 1.5 g/L, and MgSO4·7H2O 0.2 g/L; pH 7 ± 0.2, the solid medium was made by the addition of 2% agar powder, all the above reagents are analytically pure and purchased from Aladdin Biochemical Technology Co . Citric acid-disodium hydrogen phosphate buffer , polyethylene glycol 20,000 were purchased from Sangon Biotech , Sephadex G-75 column were purchased from Macklin .Triglyceride medium: peptone 10 g/L, yeast extract 5 g/L, NaCl 5 g/L, triglyceride 2 mL/L, and agar 20 g/L. LB medium: Peptone 10 g/L, yeast extract 5 g/L, NaCl 10 g/L, inorganic salt medium (MSM): CH2O 9.5 μL. The final concentration of PCR reaction reagent was 0.4 μM. Finally, the PCR products were submitted to Sangon Biotech  for nucleotide sequencing, the sequencing results were compared with the 16S rRNA sequences on NCBI by BLAST. Nucleotide sequences of strains close to their homology were obtained from GenBank, and the sequence of ZF2 was repeatedly compared with other sequences using the Clustal W comparison method. Software MEGA 11.0. was used to build a phylogenetic tree by neighbor-joining (NJ) methods.As a source of microorganisms, samples were collected from pig carcasses and sawdust compost during the high temperature period on day 10 (when temperatures were as high as 56.3 ℃) and transported immediately to the laboratory. The compost sample information  is shown in Zhai . About 1600 reached 1.0. 1% (v/v) cultures were inoculated into a 150 mL conical flask containing 100 mL MSM and incubated at 37 ℃, 120 rpm for 48 h. The cultures were centrifuged  and the supernatant was collected as crude lipase. Lipase activity was defined as the substrate hydrolysis in 1 min produces 1 μmol titratable fatty acid, which is 1 U/g (U/mL) [Individual colony was inoculated in 5 mL MSM, cultured in a shaker at 37 ℃, 120 rpm until the ODg (U/mL) . In thisTo optimize the production of lipase, the medium composition and culture conditions were gradually optimized. In the optimization experiment, each treatment group was replicated three times, and all of them were incubated for 2 d according to the conditions of 1% inoculum, 37 ℃, and 120 r/min.3COONa in the fermentation medium as carbon sources, respectively, and the enzyme activities were determined. After the optimal carbon source selection is completed, the carbon source concentration was adjusted to 1, 3, 5, 7, and 9 g/L, and the enzyme activities were measured to determine the optimal carbon source concentration. NH4Cl in the medium was replaced with (NH4)2SO4, NaNO3, urea, peptone, and yeast extract as the nitrogen source, and the enzyme activities were determined. After determining the optimal nitrogen source, the nitrogen source was adjusted to 1, 3, 5, 7, and 9 g/L, and the enzyme activity was measured to determine the optimal nitrogen source concentration. In addition to the optimal carbon and nitrogen sources, Cu2+, Mn2+, Mg2+, Zn2+, Fe3+, Fe2+, and Ca2+ were added at a 1 mmol/L final concentration to determine the enzyme activities. After selecting the optimal ion, ion concentrations were adjusted to 0.5, 1.0, 1.5, 2.0, and 2.5 mmol/L to determine enzyme activities, based on enzyme activity, the optimal concentration was selected.Sodium citrate, sucrose, glucose, maltose, and sodium carboxymethyl cellulose were used to replace CHBased on the optimized fermentation medium, the culture was carried out according to the initial pH of 4, 5, 6, 7, 8, and 9, the inoculum amount of 1%, 3%, 5%, 7%, and 9%, the incubation temperatures of 20, 25, 30, 35 and 40 ℃, and the incubation times of 12, 24, 36, 48, 60 and 72 h, respectively. Subsequently, the enzyme activities were measured and the optimal culture conditions were selected according to the enzyme activities.2 until no BaSO4 was produced. The solution was concentrated with polyethylene glycol 20,000 and filtered through a 0.22 filter membrane. Finally, the lipase activity and protein concentration in the supernatant and precipitate were measured to select the optimal ammonium sulfate salting interval. Subsequent experiments were performed with the optimal ammonium sulfate salting interval.20 mL of the crude enzyme solution was added to eight 50 ml beakers, respectively, and solid powdered ammonium sulfate was added under ice bath conditions and stirred until saturation was 10, 20, 30, 40, 50, 60, 70, and 80%, respectively. Refrigerate at 4 ℃ overnight. Then centrifuge at 12,000 r/min for 25 min at 4 ℃. The precipitate was redissolved with 10 mL citric acid-disodium hydrogen phosphate buffer and dialyzed by using a dialysis bag (14 KDa). The dialysate was changed every 4 h and tested with BaClThe lipase was further purified by Sephadex G-75 gel exclusion chromatography. The column was pre-equilibrated with citric acid-sodium hydrogen phosphate buffer and detected by a 280 nm UV detector. When the value stopped fluctuating, equilibration was completed. The elution flow rate was adjusted to 0.5 mL/min, and one tube was collected every 3 mL, the enzyme activity was measured in each tube. Finally, molecular weights were estimated by SDS-PAGE, proteins were separated with 12% (W/V) acrylamide, and molecular weight was assessed with the protein molecular mass marker . Gels were stained with 0.25% Komas blue (R-250) and decolorized in 1% acetic acid.The tape samples were prepared and lyophilized, and 40 μl of trypsin buffer was added and incubated at 37 °C for 16-18 h. The mobile phase A was 0.1% formic acid aqueous solution and B was 0.1% formic acid acetonitrile aqueous solution. The liquid chromatography column  was equilibrated with 95% A solution, the samples were loaded by an autosampler into Zorbax 300SB-C18 peptide traps , and separated in a liquid chromatography column with a liquid phase gradient: 0–15 min, linear gradient from 4 to 50% for B solution; 15–19 min, linear gradient from 50 to 100% for B solution; 19–20 min, liquid B was maintained at 100%. Subsequent mass spectrometry was performed using a mass spectrometer Q Exactive, analysis time: 20 min, detection: positive ions. The software MaxQuant 1.5.5.1 was used to search the corresponding database for raw files of mass spectrometry assays, and the protein identification and quantitative analysis results were obtained. Based on the identification results, primary structure analysis and secondary and tertiary structure prediction, homology modeling, and crystal structure analysis were performed on the proteins.+, K+, Li+, Co+, Ca2+, Cu2+, Mg2+, Mn2+, Fe2+, Zn2+ and Fe3+, 5% and 10% ethanol, isopropanol, DMSO, methanol, n-hexane, and toluene, SDS, Tween 20, Tween 80, gum arabic, and Triton-x-100, respectively. The heat stability was also examined and enzyme activity was determined.To investigate the effect of different factors  on lipase activity, the enzyme activity of the control group was set to 100%. The experimental groups of lipases were incubated at 30–80 °C, 1 mM and 5 mM AgTo study the action of lipase on different substrates, the substrates were replaced by animal fats  and vegetable fats . The enzyme activity was defined as 100% when olive oil was used as the substrate, and this was used as a basis to determine the lipase activity with other substrates. All animal and vegetable fats and oils are commercial kitchen oils.p < 0.05); ** (p < 0.01). 0.01 *** (p < 0.001)).The resultant graphs were plotted using GraphPad Prism 9.0 and statistical analyses were performed using SPSS 22.0, with one-way ANOVA selected for significance analysis. Data points are means of three experiments and error bars represent standard errors. 2SO4 under different nitrogen sources 2SO4, and Mn2+ were 3 g/L, 7 g/L, and 1.0 mmol/L, respectively, under which the enzyme activity could reach 18.64 U/mL, 26.31 U/mL, and 26.19 U/mL, respectively , the lipase activity decreased, so the optimal inoculum was determined to be 5%  was achieved Fig. B. The efe and pH . High tePseudomonas putida 922 was 48 h [Pseudomonas aeruginosa JCM5962 was 4 days [2+ 1.0 mmol/L, inoculum volume 5%, initial pH 6, temperature 25 ℃, and fermentation time 48 h.In general, extracellular lipases are produced during the logarithmic late or stable phase of microbial culture . The effwas 48 h ; the opts 4 days , the difp < 0.05), indicating that after the lipase was completely precipitated, the precipitated of other proteins affected the enzyme activity. Therefore, 40% to 70% ammonium sulfate saturation was used to precipitate the crude lipase . The results showed that the lipase has 90.4% homology with the extracellular chitosanase produced by Bacillus subtilis 168 (Entry O07921) , the results are shown in Table 1199H1875N341O387S5, and the theoretical isoelectric point is 5.44. The instability index (II) of the protein is 39.63 less than 40, indicating that it is a stable protein [The molecular chemical formula, isoelectric point, and molecular mass of the protein were analyzed using ProtParam software , the results are shown in Fig. Shttp://npsa-pbil.ibcp.fr/cgi-bin/npsa_automat.pl?page=npsa_sopma.html), and the results are shown in Fig. Sα-helix (53.75%), 12 amino acids were β-folded (5.00%), 27 amino acids were extended chain (11.25%) and 72 amino acids were irregularly coiled (30.00%).The amino acid composition of the protein is shown in Table Sα-helix depression exists in the structure. The protein electrostatic surface is shown in Fig. https://swissmodel.expasy.org/interactive/), and its computational structural model and crystal structure after purification are shown in Fig. The Rasch analysis plot of the protein homology model is shown in Fig. https://pubchem.ncbi.nlm.nih.gov/) for molecular docking with protein models in the Molecular Operating Environment (Autodock vina 12.0) platform, and the results are shown in Table The palmitic acid was selected in the Pubchem database  and Glu203 (A). The lipase was heat resistant and less affected by metal ions, organic solvents, and surfactants, and the lipase also showed degradation ability for different animal and vegetable fats. In short, lipase has the potential to degrade kitchen waste oil, future efforts will be devoted to the use of lipase with different enzyme preparations and the immobilization of lipase aiming to contribute to food and industrial waste treatment.In this study, a lipase producing strain of Additional file 1.


Vibrio sp. LA is a strain capable of producing cold-adapted laccases. The laccase coding gene L01 was cloned from this strain and expressed in Yarrowia lipolytica, a host with good secretion ability. The secreted L01  had the activity and specific activity of 18.6 U/mL and 98.6 U/mg toward ABTS, respectively. The highest activity occurred at 35 °C. At 20 °C, L01 activity was over 70% of the maximum activity in pH conditions ranging from 4.5–10.0. Several synthetic dyes were efficiently degraded by L01. Owing to its robustness, salt tolerance, and pH stability, L01 is a promising catalytic tool for potential industrial applications.Laccases have been widely used for fruit juice clarification, food modification, and paper pulp delignification. In addition, laccases exhibit remarkable performance in the degradation of toxic substances, including pesticides, organic synthetic dyes, antibiotics, and organic pollutants. Thus, the screening and development of robust laccases has attracted significant attention. In this study,  Laccases EC 1.10.3.2), a class of polyphenol oxidases, are promising biocatalysts that are independent of cofactors .10.3.2, .Laccases are already used in large scale in the textile industry. Together with low molecular weight redox-mediator compounds, laccases can generate a desired worn appearance on denim by bleaching indigo dye ,3. The pLaccases from different organisms have been purified and characterized. The sources of these laccases include plants, insects, fungi, and bacteria . The genBacillus sp., Arthrospira sp., Achromobacter sp., Citrobacter sp., Sinorhizobium sp., and Klebsiella sp. [Vibrio sp. LA. The laccase coding gene L01 was cloned from genomic DNA and expressed in Yarrowia lipolytica. Recombinant laccase L01 is pH-stable and salt-tolerant; thus, it is an efficient tool for catalysis in industrial production.Laccases from bacteria generally possess a wide and suitable pH range and a large yield. Laccase-producing strains have been found in ella sp. . In this2+ at 25 °C. With 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as the substrate, the strain LA was detected with the highest laccase activity of 0.82 ± 0.07 U/mL at flask level (data not shown). For identification, the strain LA was subjected to 16S rDNA sequencing. BLAST alignment showed that the 16S rDNA sequence had the highest similarity (98.05%) to that of an identified strain of Vibrio algivorus, indicating that strain LA is a member of the genus Vibrio. Thus, the assigned designation was Vibrio sp. LA.Laccases are mainly found in terrestrial bacteria and fungi. To date, few marine bacteria have been shown to produce laccases ,7,8. ConL01 gene encoding laccase secreted by Vibrio sp. LA, the genomic DNA of this strain was sequenced and annotated. The putative laccase-encoding gene was confirmed by catalytic region analysis. The coding sequence (CDS) starting from ATG contained 1383 bp, with a translated protein consisting 460 amino acids. It was found that the protein has a theoretical isoelectric point (pI) of 6.14 and a calculated molecular weight (MW) of 51,600 Da. In L01, the first 25 amino acids were inferred to be the signal peptide, which agreed with the extracellular location of L01. NCBI BLAST analysis revealed that L01 has a conserved domain and is a multicopper oxidase.The laccase from strain LA was named L01. To obtain the Streptomyces ipomoeae (WP 141595389.1) and Ochrobactrum sp. 531 (ADK66266.1) [Campylobacter jejuni (PDB: 3ZX1) as the model (Fusarium oxysporum (KAG7428168.1), Bacillus amyloliquefaciens (QHT73050.1), Aspergillus niger (GAQ36284.1), Bacillus subtilis (AEK80414.1), Ochrobactrum sp. (ADK66266.1), and Aureobasidium pullulans (AUW14931.1). The results reflect that L01 has typical conserved motifs appearing in laccases, such as “HPXHXHGHXF”, “WYHPH”, “HWHGI”, “HPXHXHGHXF”, and “HCH”  ,11. The he model b, with and “HCH” , which amoeae WP 1595389.1m KAG74288.1, BaciE. coli, which is infeasible for their industrial production because of the weak secretory ability of E. coli and the complex post-processing procedures of laccases [L01 gene in Yarrowia lipolytica, a commonly used heterologous host that can produce extraordinary extracellular secretion. Many industrial enzymes have been expressed in this host with high extracellular activities [In previous studies, laccases were mostly expressed in laccases ,7,17. Intivities ,19,20,21We investigated the enzymatic properties of purified L01 and found that its catalytic activity peaked at 35 °C toward ABTS, decreased dramatically above 50 °C, and was 51.2 ± 2.1% and 74.5 ± 5.2% of the maximum activity at 10 °C and 20 °C, respectively a. L01 waMost bacterial laccases exhibit their highest activity below 40 °C. Most laccases have an activity of less than half of the highest level below 30 °C ,25,26,27The optimal pH for the activity of L01 toward ABTS was 8.0, and its activity in the range of pH 4.5–10.0 was higher than 60% of the maximum a. After Aureobasidium melanogenum and Pseudomonas extremorientalis are stable in narrow pH ranges of 2.8–3.6 and 7.0–10.0, respectively [Aquisalibacillus elongatus [A. elongatus in terms of pH stability, which can contribute to the biotransformation of various phenolic substrates, especially in cases with a significant pH change.Generally, fungal laccases play a catalytic role in oxidation reactions within a narrow acidic pH range, whereas bacterial laccases prefer a relatively neutral pH ,13,14,15ectively ,24. It ilongatus . L01 out+ were 1 mM, 10 mM, and 20 mM, the relative activities were 106.8 ± 2.3%, 119.2 ± 3.1%, and 121.5 ± 3.2%, respectively, compared to that of the control. With the presence of Na+ at 10 mM or 20 mM, the relative activities were also elevated to 126.3 ± 0.9% and 123.2 ± 2.6%, respectively. Among the tested metal ions, Fe2+ and Ca2+ were detrimental to L01 activity, reducing L01 activity to 41.7 ± 1.0% and 56.1 ± 2.1% at 20 mM, respectively  was used to produce the recombinant enzyme [2216E plates were used to isolate bacteria from seawater samples . The aux2+ at 25 °C for 48 h to screen for potential laccase-producing strains [2+ at 25 °C. PCR amplification was performed on the 16S rDNA of strain LA using universal primers 27F (5′-AGAGTTTGATCCTGGCTCAG-3′) and 1492R (5′-TACCTTGTTACGACTT-3′). 16S rDNA was then subjected to sequencing and BLAST alignment. After centrifugation of the culture at 5000× g, laccase activity of the supernatant was determined using 190 μL of 1 mM ABTS solution  [The isolated strains were cultivated on 2216E plates in the presence of 0.4 mM Cu strains . A total pH 4.5) . The reaL01 gene, the genomic DNA of LA was sent to Novogene (China) for sequencing and annotation. A putative laccase-encoding gene was identified. We analyzed the signal peptide using the online server ) and performed conserved domain analysis (https://www.ncbi.nlm.nih.gov/cdd (accessed on 11 December 2022)). The theoretical pI and Mw values were also calculated (http://web.expasy.org/compute_pi/ (accessed on 11 December 2022)). We also constructed a phylogenetic tree based on the amino acid sequences of L01 and other laccases, using MEGA version 7.0. Multiple sequence alignments were performed on L01, and laccases were characterized using DNAMAN 6.0.To obtain the L01 with the XPR2 signal peptide after codons were optimized, and then transformed the obtained DNA fragment into a URA-strain [We entrusted Synbio Technologies  to synthesize the gene A-strain . The traTo clarify the influence of temperature on L01 activity, we investigated a hydrolysis reaction catalyzed by L01 in glycine-NaOH buffer  at different temperatures (10–60 °C). The activity of L01 was used to determine the optimal reaction temperature. To clarify its thermal stability, we incubated purified L01 for 2 h at 10–60 °C and then detected the remaining activity at 35 °C. ABTS solutions with substrates were prepared with different pH buffers  and Na2HPO4-citric acid ), which were used in enzymatic activity assays to determine the optimal pH for the reaction. The pH stability was estimated by detecting enzymatic activity after 4 h of incubation at 4 °C in pH buffers. The reactions were performed in triplicate.High-concentration metal ions and compounds were diluted with ABTS solution to a concentration of 1 mM. L01 activity was detected in the presence of various metal ions and some compounds through reactions catalyzed by L01 at 35 °C. The reactions were performed in triplicate.Aspergillus oryzae , with an activity of 5 U/mL was added to the dye solution. The resulting mixtures were incubated for 6 h and 24 h at 28 °C. The ODs at the maximum absorbance wavelengths were recorded using a spectrophotometer. Dyes mixed with inactivated L01 were used as controls, and their ODs at maximal absorbance wavelengths were determined using the same method [Safranine 0 (60 mg/L), methylene blue (10 mg/L), bromophenol blue (300 mg/L), eriochrome black T (300 mg/L), crystal violet (10 mg/L), malachite green (10 mg/L), and methyl orange (10 mg/L) were prepared in a pH 5.0 solution. L01 solution and commercial laccase from e method ,32. The In this study, a new cold-adapted laccase was cloned, extracellularly expressed, and characterized. Recombinant laccase L01 presented its highest activity at 35 °C and retained over 50% of its maximum activity at 10 °C and 20 °C. In addition, over 60% of the activity was maintained at pH 4.5–10.0. In addition to being pH-stable, L01 was salt-tolerant and NaCl-independent. Several synthetic dyes were efficiently degraded. L01 is an attractive biocatalyst for industrial applications owing to its robustness and unique pH stability.


Matricaria chamomilla flower essential oils (EOs) blue Egyptian (EO-1), chamomile German CO2 (EO-2), and chamomile German (EO-3) and the pure compound α-bisabolol were evaluated against red imported fire ants (RIFA), Solenopsis invicta Buren, black imported fire ants, S. richteri Forel (BIFA), and hybrid imported fire ants (HIFA) for their repellency and toxicity. A series of serial dilutions were tested starting from 125 µg/g until the failure of the treatment. Based on the amount of sand removed, EO-1 showed significant repellency at dosages of 7.8, 7.8, and 31.25 µg/g against RIFA, BIFA, and HIFA, respectively. EO-3 was repellent at 3.9, 7.8, and 31.25 µg/g against BIFA, RIFA, and HIFA, whereas α-bisabolol was active at 7.8, 7.8, and 31.25 µg/g against BIFA, HIFA, and RIFA, respectively. DEET  was active at 31.25 µg/g. Toxicity of EOs and α-bisabolol was mild to moderate. For EO-1, LC50 values were 93.6 and 188.11 µg/g against RIFA and BIFA; 98.11 and 138.4 µg/g for EO-2; and 142.92 and 202.49 µg/g for EO-3, respectively. The LC50 of α-bisabolol was 159.23 µg/g against RIFA. In conclusion, M. chamomilla EOs and α-bisabolol offer great potential to be developed as imported fire ant repellents. Solenopsis invicta Buren), black imported fire ants  (Hymenoptera: Formicidae), and hybrid imported fire ants are notorious pests, impacting humans, wildlife, pets, and livestock through their venomous stings  non-3-ene (6.98%), 7-methoxycoumarin (2.60%), α- bisabolol oxide B (1.45%), (E)-β-farnesene (1.39%), and 9,12-octadecadienoic acid -(1.39%) as major compounds. EO-1 and EO-3 had similar major metabolites except that bisabolol oxide A was 49.19% and 27.83%, in EO-1 and EO-3, respectively.EO-M. chamomilla EOs, its selected pure compound, and the positive control DEET is presented are 1 showed significantly higher repellency than ethanol at dosages of 125–7.8 µg/g against RIFA and BIFA, whereas the activity at 62.5–31.25 µg/g was similar to ethanol against HIFA. EO-2 showed significantly higher repellency than ethanol at dosages of 125–15.6 µg/g against RIFA, whereas the activity at 15.6–3.9 µg/g was similar to ethanol against BIFA and HIFA. EO-3 showed significantly higher repellency than ethanol at dosages of 125–3.9 µg/g against BIFA, whereas the activity was similar to ethanol at 3.9 µg/g against RIFA and at 31.25 µg/g against HIFA. α-Bisabolol showed significantly higher repellency than ethanol at dosages of 125–7.8 µg/g against BIFA and HIFA, whereas the activity was similar to ethanol at 15.6–3.9 µg/g against RIFA. Against BIFA and RIFA, DEET treatments showed significantly higher repellency than ethanol at dosages of 125–62.5 µg/g, whereas the repellency at 31.25 µg/g was similar with ethanol. In HIFA, the repellency of DEET was significantly higher at dosages of 125–62.5 µg/g, whereas the repellency at 31.25 µg/g was similar to ethanol.Mean weight (g) of sand removed by the workers in a digging bioassay treated with different concentrations of M. chamomilla EOs, its pure compound, and bifenthrin at 24 h post-treatment against RIFA, BIFA, and HIFA workers are given in 1 showed toxicity with LC50 values of 93.60 and 188.11 µg/g against RIFA and BIFA workers, respectively. LC50 values of EO-2 were 98.11 and 138.40 µg/g against RIFA and BIFA, respectively. EO-3 showed LC50 of 142.92 and 202.49 µg/g against RIFA and BIFA workers, respectively, whereas LC50 value of α-bisabolol was 159.23 µg/g against RIFA workers. α-Bisabolol gave 30% mortality against BIFA at the highest screening dose of 250 µg/g. EO-1, EO-2, EO-3, and α-bisabolol gave 73%, 20%, 40%, and 80% mortality at the highest screening dose of 250 µg/g against HIFA. Bifenthrin with LC50 values of 0.03, 0.032, and 0.018 µg/g against RIFA, BIFA, and HIFA workers, respectively, at 24 h post treatment was more toxic than the essential oils. Based on LC50 values, toxicity of M. chamomilla EOs and α-bisabolol was significantly higher in RIFA as compared to BIFA.Toxicity data of 1 and EO-3) due to the presence of chamazulene, which originates from pro-azulenes (matricin and matricarin) at the increased temperatures and in the presence of organic acids contained in the flowers , and at the dose of 1 μL/1 L of air, M. recutita essential oil induced a 70.7% and 60.1% reduction in oviposition and emergence [M. chamomilla at a concentration of 15% caused significant mortality (LC50 = 7.78%) in adults T. castaneum at 24 h post-treatment [50 values, EO-1 and EO-3, and α-bisabolol, were more toxic to RIFA as compared to BIFA. The dose–response curves could not be developed for HIFA because all the samples showed less than 100% mortality at the highest screening dose of 250 µg/g and abruptly dropped to 0 % at the next serial dose at 24 h post-treatment. Still, the percent mortality data from this study can be used to predict the toxicity potential of EOs and the pure compound from M. chamomilla against HIFA workers. This is the first detailed report of the toxicity of M. chamomilla EOs and its pure compound against imported fire ant workers.Insecticidal activity of  2 weeks . Wheat gexposure . EOs of mergence . EOs frotality LC = 7.78% 2 essential oil (EO-2) and chamomile German essential oil (EO-3) were purchased from Eden’s Garden . α-Bisabolol was purchased from Sigma Aldrich . GC-MS analysis of three EOs was performed in Agilent 7890 B GC system, which was equipped with a 5977A quadrupole mass spectrometer and with a 7693 autosampler . Samples were prepared at 10 mg/mL concentration in GC-MS-grade methylene chloride . The carrier gas Helium at a constant flow rate of 1.5 mL/min was used. The inlet temperature in split injection mode was set to 280 °C, with a 30:1 split ratio. The initial oven temperature program was set at 60 °C for 2 min, and was increased to 280 °C at a rate of 6 °C/min. Isothermal was at 280 °C for 10 min. The overall run time was 77 min. Agilent MassHunter software (version B.07.06) was used to perform data acquisition.Chamomile oil Blue Egyptian (EO-1) was purchased from Perfumer Supply House  and chamomile German CORIFA, BIFA, and HIFA workers were used in these bioassays. BIFA colonies were brought from Tunica County, MS-713, Hernando, MS 38632 (34°49′56.5″ N 90°12′55.6″ W). RIFA collection was made from Washington County, MS 38748 (33°9′31.2″ N 90°54′56.4″ W). HIFA workers were used from the mounds located under natural field conditions at University Field Station . Ant collection was brought to the laboratory and kept in plastic trays, whose top inner walls were coated with Insect a Slip  to prevent the escape of ants. Crickets and 25% honey-water solution were available as food to the ants. A test tube filled with water and plugged with cotton served as a water source. Moist sand-filled 45 mL fluted aluminum dishes  were kept inside trays to serve as a digging substrate. The ants were maintained under laboratory conditions of 32 ± 2 °C temperature and 50 ± 10% relative humidity for one month before starting the bioassays. The ant species were identified based on venom alkaloid and hydrocarbon indices of the collected individuals ,56.M. chamomilla, a pure compound, and a positive control DEET  were tested. A series of dilutions were tested starting from 125 µg/g until the failure of the treatment. Overall, three sets of the replicates were run on 3 different days. Sand removal data were analyzed by Analysis of Variance (ANOVA)  and means were separated by using Ryan–Einot–Gabriel–Welsch multiple range test ).According to Deletre et al. , repelle50 values were calculated by using probit analysis (SAS 2012).The toxicity bioassay described by Ali et al.  was usedM. chamomilla against red, black, and hybrid imported fire ants. Removal of treated sand in digging bioassays, which may directly be proportional to inhibition of digging ability of the fire ants, was used as a criterion for repellency. Based on the above criterion, essential oils and α-bisabolol showed the potential to be significant repellents against the imported fire ants. All the natural products tested in this study, especially EO-1, EO-3, and α-bisabolol showed repellency higher than DEET. Further studies will be conducted to explore repellent/toxicant potential of these essential oils and the pure compound by testing in different formulations against imported fire ants in elaborate laboratory and field trials.In conclusion, this is the first report on the insecticidal and repellent activities of essential oils of


A field emission scanning electron microscope (FE-SEM), Fourier transform infrared spectroscopy (FT-IR), and X-ray diffraction were used to characterize the synthesized products. The FE-SEM images showed that the analcime and Fe3O4 samples consist of polyhedral and quasi-spherical shapes with average diameters of 923.28 and 28.57 nm, respectively. Besides, the Fe3O4/analcime nanocomposite consists of polyhedral and quasi-spherical shapes with average diameters of 1100.00 nm. The greatest uptake capability of the Fe3O4/analcime nanocomposite toward the copper and cadmium ions is 176.68 and 203.67 mg/g, respectively. The pseudo-second-order kinetic model and Langmuir equilibrium isotherm best describe the uptake of copper and cadmium ions using the Fe3O4/analcime nanocomposite. The uptake of copper and cadmium ions using the Fe3O4/analcime nanocomposite is exothermic and chemical in nature.In the water purification field, heavy metal pollution is a problem that causes severe risk aversion. This study aimed to examine the disposal of cadmium and copper ions from aqueous solutions by a novel Fe The quenching constant of nanocomposite with picric acid was 4.30 × 104 M−1 in DMSO with a limit of detection up to 2.20 μM as a consequence of turn off sensing [3N4/Fe2O3 nanocomposite for photocatalytic removal of indoor formaldehyde under visible light [2O3/HNb3O8 nanocomposite for the uptake of ethyl mercaptan in methane gas where the maximum uptake capability equals 48.05 mg/g [3O4/analcime nanocomposites to remove copper and cadmium ions. The feature of Fe3O4/analcime nanocomposite is its ability to remove copper and cadmium ions using the ion exchange adsorption process, and hence the adsorption capacity is expected to be great. The FE-SEM, XRD, and FT-IR techniques were used to illustrate the structural and physical properties of the synthesized adsorbent. Moreover, the effects of temperature, pH, time, and concentration on the uptake of copper and cadmium ions from aqueous media were examined.In recent decades, water contamination has been one of the greatest significant environmental problems that humans have faced. Consequently, it has garnered the interest of numerous academics and scientists –3. Inorgar level . The cryar level . In additoxicity –28. Also.35 mg/g . Kumar e sensing . Wang etle light . Wang et.05 mg/g . Consump.05 mg/g . Deng et.05 mg/g . Kuang e.05 mg/g . Yang et.05 mg/g . Alvarez.05 mg/g . Joseph .05 mg/g . The obj4·7H2O), sodium hydroxide (NaOH), iron(III) chloride hexahydrate (FeCl3·6H2O), aluminum chloride hexahydrate (AlCl3·6H2O), cadmium(II) nitrate tetrahydrate (Cd(NO3)2·4H2O), glutamine (C5H10N2O3), copper sulfate (CuSO4), hydrochloric acid (HCl), ammonium hydroxide (NH4OH), potassium nitrate (KNO3), ethylenediaminetetraacetic acid disodium salt dihydrate (C10H14N2Na2O8·2H2O), and fumed silica (SiO2) were obtained from the Sigma Aldrich Chemical Company then utilized without purifying. Besides, the analcime was prepared according to the procedure described by Youssef et al. [3O4 was prepared according to the procedure described by Kang et al. [Iron(II) sulfate heptahydrate . FT-IR spectra of the synthesized powders were gotten in the range from 4000 to 400 cm−1 using a Nicolet iS50 spectrophotometer pressed into potassium bromide. The morphology of the synthesized powders was investigated using a field emission scanning electron microscopy of model JSM-IT800 Schottky. The concentrations of copper and cadmium ions were estimated utilizing a Perkin Elmer-3300 multi-element atomic absorption spectrometer. The magnetic properties of the synthesized composite were determined at room temperature (27 °C) using a vibrating sample magnetometer (VSM-Cryogenic Limited PPMS). The stability of the synthesized composite was studied under nitrogen atmosphere using a thermal gravimetric analyzer (TGA-Shimadzu DT-60H) with a heating speed of 10 °C/min.The X-ray diffraction patterns of the chemically synthesized powders were gotten utilizing a D8 Advance X-ray diffractometer with a copper anode , time (5.00–120.00 min), uptake temperature (298.00–328.00 K), and concentration (100.00–300.00 mg/L).The batch adsorption method was used to evaluate the adsorption processes of copper and cadmium ions utilizing the Fe3O4/analcime nanocomposite (Q) were determined using Eqs. o is the initial concentration of the cadmium or copper ions (mg/L), Ceq is the equilibrium concentration of the cadmium or copper ions (mg/L) and m is the dry mass of the Fe3O4/analcime nanocomposite (g). Besides, V is considered the taken volume of the cadmium or copper solution (L).The uptake percentage of the copper or cadmium ions (% R) and the uptake capability of the FePZC) of the Fe3O4/analcime nanocomposite was estimated, as illustrated by Khalifa et al., as the following [3O4/analcime nanocomposite were mixed with 70.00 mL of about 0.03 M KNO3 solutions. Moreover, the primary pH (pHi) of the KNO3 solutions was examined in the range from 2.00 to 12.00. Each adsorbent/KNO3 blend was magnetically stirred for 9 h. The final pH values (pHf) were measured and plotted against the initial pH values (pHi). The pHpzc is the pHf level where a characteristic plateau was obtained.The point of zero charge  of cadmium or copper solution. After that, the resulting mixture was magnetically stirred for 80 min. Finally, the nanocomposite was separated using centrifugation then the concentration of copper or cadmium ions in the filtrate was determined at 193.70 and 324.70 nm using a Perkin Elmer-3300 multi-element atomic absorption spectrometer, respectively. To regenerate the nanocomposite, loaded copper or cadmium ions is completely desorbed from the surface of the nanocomposite by stirring the nanocomposite loaded with copper or cadmium ions in 50 mL of 1 M ethylenediaminetetraacetic acid disodium salt dihydrate for 30 min. Afterward, the regenerated nanocomposite was used for the removal of copper and cadmium ions for four consecutive cycles as previously described.The adsorption was carried out as the following; 0.05 g of the Fe3O4, and Fe3O4/analcime nanocomposite, respectively. Moreover, the FT-IR bands of the analcime, which were appeared at 444, 661, and 784 cm−1, are due to the bending vibrations, internal symmetric vibrations, and external symmetric vibrations of W–O–W (W=Si and/or Al), respectively. Also, the FT-IR of the analcime, which were appeared at 1006, 1415, 1648, and 3452 cm−1, are because of the internal asymmetric vibrations of W–O–W, external asymmetric vibrations of W–O–W, bending vibrations of H–O–H, and stretching vibrations of H–O–H, respectively. The FT-IR bands of the Fe3O4, which were appeared at 487 and 551 cm−1, are because of the stretching vibrations of Fe–O. Furthermore, the FT-IR bands of the Fe3O4, which were appeared at 1629 and 3422 cm−1, are due to the bending and stretching vibrations of H–O–H, respectively. The FT-IR bands of the analcime in the Fe3O4/analcime nanocomposite were observed at 440, 660, 790, 1011, 1414, 1625, and 3426 cm−1. Moreover, the FT-IR bands of the Fe3O4 in the Fe3O4/analcime nanocomposite were observed at 490 and 560 cm−1 [3O4, and Fe3O4/analcime, the results displayed that almost all the bands of the analcime and Fe3O4 were observed in the synthesized Fe3O4/analcime nanocomposite. Hence, this indicating the successful synthesis of Fe3O4 modified with analcime.Figure 560 cm−1 . Compari3O4, and Fe3O4/analcime nanocomposite, respectively. In addition, the XRD peaks, which appeared at 2Ɵ = 15.83°, 18.39°, 24.33°, 25.99°, 30.64°, 31.90°, 33.25°, 35.88°, 37.03°, 40.49°, 47.84°, 48.68°, 52.46°, 53.29°, 54.25°, 57.71°, 62.75°, 65.90°, and 69.04°, are due to the (211), (220), (321), (400), (332), (422), (431), (521), (440), (532), (640), (633), (732), (800), (741), (831), (921), (932), and (772) miller indices of the analcime (Na(AlSi2O6)(H2O)) as clarified from JCPDS No. 01-070-1575. The XRD peaks, which appeared at 2Ɵ = 30.22°, 35.57°, 43.23°, 53.83°, 57.29°, and 62.85°, are due to the (220), (311), (400), (422), (511), and (440) miller indices of the Fe3O4 as clarified from JCPDS No. 85-1436. Also, the intensity of the XRD peaks of the Fe3O4/analcime nanocomposite was affected, but it is similar to that of the analcime. In the XRD pattern of nanocomposite, the most intense peak of Fe3O4 at 2Ɵ = 35.57° was found to be diminished due to combination of Fe3O4 with analcime that have more intense peaks than Fe3O4. The mean crystallite sizes of the analcime, Fe3O4, and Fe3O4/analcime nanocomposite, which were determined using Scherrer equation, are 80.24, 15.27, and 98.29 nm, respectively [Figure ectively .Fig. 2Th3O4, and Fe3O4/analcime nanocomposite, respectively. In addition, the results showed that the analcime and Fe3O4 samples consist of polyhedral and quasi-spherical shapes with average diameters of 923.28 and 28.57 nm, respectively. Besides, the Fe3O4/analcime nanocomposite consists of polyhedral and quasi-spherical shapes with average diameters of 1100.00 nm. Upon incorporation of the Fe3O4 nanoparticles into the analcime framework, a distortion of the analcime's original polyhedral structure was observed. This suggests a bonding interaction between analcime and Fe3O4 nanoparticles, resulting in morphological changes. It is known that the SEM shows the shape of the surface of the samples, not the shape of the particles, which could be a combination of multiple particles. Therefore, SEM cannot be used to determine whether the sample is nanosized or not. XRD determines whether the sample is nanosized or not.Figure 3O4/analcime nanocomposite. The results displayed that the Fe3O4/analcime nanocomposite consists of Fe, Si, Al, O, and Na with weight percentages equal to 18.76, 12.82, 12.57, 51.10, and 4.75%, respectively. Hence, this is a confirmation that there is a bonding interaction between analcime and Fe3O4 nanoparticles.Figure The plot of magnetization of the synthesized nanocomposite at 27 °C versus magnetic field is displayed in Fig. 3O4/analcime nanocomposite, respectively. The results showed that both the uptake percentage and the uptake capability increased significantly with the increase of the pH from 2.50 to 6.50. In addition, the increase was very slight when the pH was increased from 6.50 to 8.00 owing to the constant amount of the active sites of the adsorbent. Therefore, pH 6.50 is the ideal value at which the highest uptake rate of copper and cadmium ions is achieved. At pH 6.50, the uptake percentage of copper and cadmium ions using the Fe3O4/analcime nanocomposite is 85.56 and 98.50%, respectively. Besides, the greatest uptake capability of the Fe3O4/analcime nanocomposite toward copper and cadmium ions is 167.11 and 197.00 mg/g, respectively. To understand the influence of pH on the adsorption processes of the cadmium and copper ions, the point of zero charge (pHPZC) of the Fe3O4/analcime nanocomposite was determined and found to be 3.40, as shown in Fig. PZC), the number of positively charged sites of the Fe3O4/analcime nanocomposite increased, which did not favor the uptake of positively charged copper or cadmium ions owing to electrostatic repulsion [PZC), the number of negatively charged sites of the Fe3O4/analcime nanocomposite increased, which favor the uptake of positively charged copper or cadmium ions owing to electrostatic attraction [Figure epulsion , 45.At htraction , 45. At 3O4/analcime nanocomposite, respectively. The results showed that both the uptake percentage and the uptake capability increased significantly with the increase of the time from 5.00 to 80.00 min. In addition, the increase was very slight when the disposal time was changed from 80.00 to 120.00 min due to the saturation of the active sites of the adsorbent. Therefore, 80.00 min is the ideal value at which the greatest uptake rate of copper and cadmium ions is achieved. At time equals 80.00 min, the uptake percentage of copper and cadmium ions utilizing the Fe3O4/analcime nanocomposite is 85.00 and 98.50%, respectively. Besides, the greatest uptake capability of the Fe3O4/analcime nanocomposite toward copper and cadmium ions is 170.00 and 197.00 mg/g, respectively. Two kinetic models, pseudo-first-order , Qe is the quantity of copper or cadmium ions adsorbed at equilibrium (mg/g), k1 is the pseudo-first-order rate constant (1/min), and k2 is the pseudo-second-order rate constant (g/mg.min). The kinetic constants were presented in Table 2) compared with the pseudo-second-order model. Consequently, the pseudo-second-order model may best describe the uptake of copper or cadmium ions using the Fe3O4/analcime nanocomposite.Figure 3O4/analcime nanocomposite, respectively. The results showed that both the uptake percentage and the uptake capability decreased significantly with the increase of the temperature from 298.00 to 328.00 K. Therefore, 298.00 K is the ideal value at which the greatest uptake rate of copper and cadmium ions is achieved. Exploiting Eqs. o, KJ/mol), change in enthalpy , and change in entropy , can be determined [d versus 1/T were presented in Fig. d is the distribution constant (L/g) whereas R is the universal gas constant (KJ/mol kelvin). T is the uptake temperature (kelvin). The uptake of cadmium and copper ions using the Fe3O4/analcime nanocomposite is exothermic, as revealed by the negative sign of △Ho values. Besides, the values of △Ho are higher than 40 kJ/mol, revealing that the adsorption is chemical in nature. Entropy is a measure of randomness, and its positive value in the case of the disposal of copper and cadmium ions using the Fe3O4/analcime nanocomposite means that these ions move randomly in the solution and go to the adsorbent from all directions, and thus the separation efficiency increases. The negative values of △Go revealed that the uptake of copper and cadmium ions occurred spontaneously.Figure termined , 45. The3O4/analcime nanocomposite, respectively. The results showed that the uptake percentage decreased significantly whereas the uptake capability increased significantly with the increase of the concentration from 100.00 to 300.00 mg/L. Two equilibrium isotherms, Langmuir . In addition, kF is the equilibrium constant of the Freundlich isotherm (mg/g)(L/mg)1/n whereas Qmax is the maximum uptake capability of the Langmuir isotherm (mg/g). Equation max using the Freundlich isotherm [Figure muir Eq.  and Freumuir Eq. , were apisotherm , 45.10\d2) compared with the Langmuir isotherm. Consequently, the Langmuir isotherm may best describe the disposal of copper or cadmium ions using the Fe3O4/analcime nanocomposite. The greatest uptake capability of the Fe3O4/analcime nanocomposite toward the copper and cadmium ions is 176.68 and 203.67 mg/g, respectively.The equilibrium constants were presented in Table 3O4/analcime nanocomposite was superior to that of many other adsorbents that published in the literature, as exhibited in Table The uptake capability of the Fe3O4/analcime nanocomposite toward copper and cadmium ions did not change significantly for four consecutive cycles as shown in Fig. 3O4/analcime nanocomposite can be used many times without losing its effectiveness.Ethylenediaminetetraacetic acid disodium salt dihydrate is the head member of the family of ligands. Ethylenediaminetetraacetic acid disodium salt dihydrate is a hexadentate ligand forming highly stable five membered chelates with metal ions such as copper and cadmium ions. Hence, Ethylenediaminetetraacetic acid disodium salt dihydrate has the ability to completely desorb cadmium and copper ions from the surface of the nanocomposite. The uptake capability of the Fe3O4/analcime nanocomposite was facilely synthesized as novel nanostructures for the effective disposal of copper and cadmium ions from the aqueous solutions. The products were characterized using field emission scanning electron microscope (FE-SEM), Fourier transform infrared spectroscopy (FT-IR), and X-ray diffraction (XRD). The greatest uptake capability of the Fe3O4/analcime nanocomposite toward copper and cadmium ions is 176.68 and 203.67 mg/g, respectively.The Fe


Aim of the study: To assess the relationship between the severity of liver cirrhosis and its outcomes based on laboratory parameters, Child-Turcotte-Pugh (CTP) score, and upper gastrointestinal (UGI) endoscopy findings.Background: Cirrhosis is the end stage of chronic liver disease (CLD) and is characterised by progressive liver fibrosis and distortion of the liver architecture. It is a major cause of morbidity and mortality all over the world. Cirrhosis is compensated in the initial stages and later progresses to the decompensated stage with various complications. The CTP scoring system predicts mortality in patients with cirrhosis.Materials and methods: This retrospective study was done in the Department of Medicine and Gastroenterology of Tata Main Hospital (TMH), Jamshedpur, Jharkhand, India. It was conducted over a period of two years between 1 January 2019 and 31 December 2020, on 150 confirmed cases of cirrhosis.Results: The most common age group was 41-60 years  and the mean age ± standard deviation (SD) for all patients was 49.82 ± 11.63 years. In a total of 150 CLD cases, males were 96 (64%). The most common cause of CLD was alcohol . Based on presenting symptoms, most CLD patients presented with generalized weakness . The most common signs were icterus  and ascites . Most patients belonged to CTP class A , followed by CTP class B  and class C . The most common UGI endoscopy finding was portal hypertensive gastropathy (mild or severe) . Total deaths were 24 (16.00%), with 17 deaths (70.83%) in patients belonging to CTP class C.Conclusion: CLD is a common entity in eastern India with male preponderance and affects mostly people of the middle age group. Alcohol intake is a major cause of CLD, followed by non-alcoholic fatty liver disease and chronic hepatitis B and C. A significant rise in morbidity and mortality due to alcoholic liver disease (ALD) was observed in the study and needs urgent social and medical intervention. The incidence of ALD in our study was 50.67%. Cirrhosis is one of the most frequent causes of death due to hepatic diseases worldwide. Approximately two million deaths occur per year due to liver diseases, one million due to complications of cirrhosis and one million due to viral hepatitis and hepatocellular carcinoma . CirrhosGlobally, 1.6 billion people had chronic liver disease (CLD) in 2017, with non-alcoholic fatty liver disease (NAFLD) (60%), hepatitis B virus (HBV) (29%), hepatitis C virus (HCV) (9%), and alcoholic liver disease (ALD) (2%) being the most frequent causes. Additionally, cirrhosis contributed to more than 132 million (95% UI: 127-145) deaths worldwide in 2017, with 883,000  deaths among men and 440,000  deaths among women. This is a significant increase as compared to 1990 when the total deaths from CLD in both sexes were 899,000 . In 2017, these fatalities made up 2.4% (2.3-2.6) of all deaths worldwide, up from 1.9% (1.8-2.0) in 1990. Cirrhosis is thought to affect between 16.5 and 23.6 people per 100,000 in East Asia and Southeast Asia, respectively. According to statistics from the Global Burden of Disease survey, there were 20.7 cases of cirrhosis per 100,000 people in 2015, up 13% from the year 2000. Over the previous 20 years, the prevalence of cirrhosis has grown 1.5-2 times [Recent research has demonstrated that systemic inflammatory response syndrome (SIRS), with or without a confirmed bacterial infection, is an independent predictor of survival in critically unwell cirrhotic patients and is also linked to the emergence of problems caused by portal hypertension. In individuals with cirrhosis who develop multiorgan failure, liver function does not appear to be the primary driver of prognosis. In patients with severe or advanced cirrhosis, conventional measures for detecting SIRS lack sensitivity and specificity because of hypersplenism, hyperventilation coupled with encephalopathy, hyperkinetic circulation, and the use of beta-blockers . To predThis retrospective study was done at the Department of Medicine and Gastroenterology of Tata Main Hospital (TMH), Jamshedpur, Jharkhand. It was done over 150 confirmed cases of CLD admitted to the Department of Medicine and Gastroenterology between 1 January 2019 and 31 December 2020. The cases were diagnosed by ultrasonogram (USG) of the whole abdomen, UGI endoscopy, clinical features, and laboratory investigations. USG findings of CLD included shrunken liver with a nodular surface, ascites, and splenomegaly. Laboratory investigations suggestive of CLD were reversed albumin-globulin ratio with low albumin levels and deranged INR. Their records were retrieved from the hospital management system (HMS), a database of the patients coming to the TMH. Inclusion criteria included cases of CLD (compensated or decompensated) above the age of 18 years. Exclusion criteria included the existence of any concurrent infectious or systematic inflammatory diseases, significant haematologic disorders, thyroid dysfunction, and/or severe renal insufficiency  and non-hepatic malignant tumours and patients on warfarin, steroids, and hormone replacement therapy.Clinical findings, laboratory investigations, including complete blood counts, liver function tests, kidney function tests, serum electrolytes, coagulation parameters (prothrombin time and INR), blood sugar, autoimmune profile, hepatitis B surface antigen, anti-HCV antibody, HBV DNA quantitative, and HCV RNA quantitative (where required), USG of the abdomen, chest X-ray posteroanterior view, ascitic fluid, total leucocyte count (TLC), differential leucocyte count (DLC), cultures, serum ascites albumin gradient (SAAG), and UGI endoscopic data were all collected. The statistical analysis was completed using Statistical Package for Social Sciences  and Microsoft Excel . The Student's t-test and the chi-square test were employed in the data analysis to compare continuous and dichotomous variables, respectively. Descriptive statistics were also used as necessary. To investigate distributional differences, the data were tabulated and presented as frequency (n) and percentage (%), and the continuous parameters were presented as mean ± standard deviation (SD). The correlation of different CTP classes with outcomes was done with a one-way ANOVA method. Statistical significance was determined to be a p-value of 0.05 or less. Informed permission was acquired from participants before they were enrolled in our research.In the present study, the most common age of patients with CLD was 41-60 years , followed by 21-40 years  and 61-80 years . Only six (4%) patients were under 20 years of age. The mean age ± SD for all studied patients was 49.82 ± 11.63 years  were male and 54 (36%) were female , followed by NAFLD 37, 24.67%). Hepatitis B, hepatitis C, and cryptogenic factor-causing CLD cases were 11 (07.33%), seven (04.67%), and 15 (10.00%), respectively. The least number of cases were of autoimmune CLD  , followed by jaundice  and blood in vomiting and/or black stool . Of the patients, 49 32.67%) had abdominal distension due to ascites. Pedal oedema and altered sensorium also presented in 27 (18.00%) cases each. The least common symptom was fever  , ascites (29.33%), pedal oedema (13.33%), and asterixis (07.33%). There was an overlap of multiple clinical signs in many patients , followed by CTP class B 44, 29.33%) and class C   135, 75%) and grade I varices  , aspartate aminotransferase (AST), haemoglobin, WBC, platelets, INR, serum creatinine, and serum electrolytes in different CTP classes were evaluated and it was found that all findings were statistically significant with p-value < 0.001, but serum creatinine and serum potassium levels showed statistically insignificant results Table .The total number of deaths in the present study was 24 (16.00%). A total of 17 (58.62%) deaths belonged to CTP class C and seven deaths (18.92%) belonged to CTP class B , followed by 21-40  and 61-80 years . The mean age ± SD for all patients was 49.82 ± 11.63 years. In a study of 50 patients by Sreenivas et al., the mean age of presentation was 43.84 years with an SD of 10.4 years [.33%, folIn the present study, most CLD patients were males 96, 64%). Similar results were seen in a study by Sreenivas et al. who included 50 patients, out of which 46 (92%) were males and four 8%) were females , 64%. Si. Out of % were feIn the present study, the most common cause of CLD was alcohol 76, 50.67%), followed by NAFLD . Hepatitis B, hepatitis C, cryptogenic, and autoimmune CLD cases were 11 (07.33%), seven (04.67%), 15 (10.00%), and four (02.66%), respectively. Sreenivas et al. showed that out of 50 CLD cases, alcohol was the aetiological factor in 41 patients, HCV infection in three patients, HBV infection in one patient, alcohol and HBV in three patients, and alcohol and HCV infection in one patient, respectively , 50.67%,. Also, AIn the present study, most CLD patients presented with generalized weakness 144, 96.00%). In contrast to the present study, Sreenivas et al., who included 50 patients with CLD, showed that the most common clinical presentation was abdominal distension (94%), followed by jaundice (56%) [.00%. In UGI endoscopy findings in the present study showed overlapping findings, out of which the most common were portal hypertensive gastropathy (either mild or severe) , followed by grade I varices 63, 42.00%). In contrast to the present study, Sreenivas et al. found that out of 50 patients with CLD, most patients showed normal UGI endoscopy , followed by grade II varices  and grade IV varices , respectively [3, 42.00%The present study showed that the mean bilirubin in CTP classes A, B, and C was 02.14 ± 00.87, 02.67 ± 01.07, and 03.65 ± 01.57 md/dl, respectively; mean albumin was 03.07 ± 01.11, 02.32 ± 01.01, and 01.92 ± 00.72 gm/dl; mean ALT was 41.24 ± 10.54, 47.24 ± 09.82, and 44.11 ± 06.54 IU/L; mean AST was 47.81 ± 07.63, 58.47 ± 09.77, and 51.24 ± 10.24 IU/L; mean haemoglobin was 10.12 ± 02.12, 09.14 ± 02.54, and 08.41 ± 02.55 gm/dl; mean WBC was 07.55 ± 05.11, 08.41 ± 04.21, and 08.74 ± 03.87 x 103/ml; mean platelets level was 102.50 ± 24.12, 98.20 ± 31.24, and 78.24 ± 21.14; mean INR was 01.41 ± 00.41, 01.31 ± 00.57, and 01.87 ± 00.77; mean serum creatinine (mg/dl) was 01.21 ± 00.21, 01.31 ± 00.57, and 01.61 ± 00.65 x103/ml; mean Na was 128.54 ± 03.21, 126.41 ± 03.11, and 125.41 ± 02.41 mEq/l; and mean K was 04.41 ± 01.11, 04.25 ± 01.23, and 04.88 ± 01.57 mEq/l, respectively. The above findings were statistically significant when compared with healthy individuals with a p-value < 0.001, except for serum creatinine and serum potassium, which showed a p-value > 0.05. Similarly, Tawfik et al. [In the present study, the total number of deaths out of the 150 patients was 24 16.00%). The total number of deaths among 29 patients in CTP class C was 17  and among 44 patients in CTP class B was seven (29.16%), respectively. No deaths were seen in CTP class A. Sreenivas et al. showed that the total number of deaths was 12 (24%) [.00%. TheThe study observed 150 CLD patients visiting a tertiary care centre in eastern India over a two-year period. The most common age group was 41-60 years, and the majority of patients were male. Alcohol was the most common cause of CLD, followed by NAFLD. A significant increasing trend of ALD-related morbidity and mortality was observed in this study. The study provides demographics and clinical features of CLD patients in this group of population and underscores the need for preventive measures, early detection, and medical attention to address the burden of this condition.


Firstly, a novel approach of synthesizing of a new substituted pyridooxadiazine-7-carboxylic acid (PO) is reported through reacting(Z)-N’-(1-(3-aminophenyl)ethylidene)-2-cyanoacetohydrazide with (Z)-ethyl 2-cyano-3-(pyridin-3-yl)acrylate. Then Dialdehyde chitosan (DACs) has prepared via periodat oxidation of chitosan (Cs). The synthesized compounds have studied via various spectroscopic instruments to validate their chemical structure such as nuclear magnetic resonance 1 H NMR, 13 C NMR, fourier transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD), and scanning electron microscopy (SEM). The substituted pyrido oxadiazine and the composite were evaluated for antimicrobial activity against pathogenic bacteria and unicellular fungi. The results revealed that, the composite exhibited promising antimicrobial activity against E. coli, S. aureus, B. subtilis and C. albicans where inhibition zones were 19, 18, 36 and 20 mm respectively. Furthermore, the substituted pyrido oxadiazine and the composite were evaluated for cytotoxic activity against MCF-7 human breast cancer cell line as well as vero normal cell line. Results illustrated the prepared composite has anticancer activity against MCF7 where IC50 was 238 µg/ml.Synthesis of new compounds that have biological activity is an indispensible issue in order to deal with the drug resistant bacteria. This wok reports preparation of a novel composite based on substituted pyrido[2,1-The online version contains supplementary material available at 10.1186/s13065-023-01005-1. Multi-drug resistant bacteria emerged due to the overuse or misuse of antibiotics; additionally, the modern treatment of antibiotics via clinically prescribed dosages is not able to manage these pathogens, and therefore preventive strategies are necessary . Mortali245aluminium plates with visualization under UV light. Determination of the melting point was carried out using open capillary tubes on a Stuart SMP30 melting point apparatus and was uncorrected. Spectral data of the compounds were carried out in the Micro-analytical labs in the National Research Centre Cairo, Egypt. The NMR spectra was measured on a Bruker Fourier 500  at 300 K. FT-IR spectra were recorded in the range of 400–4000 cm − 1on (Shimadzu 8400 S) FT-IR Spectrophotometer. The surface morphology was analyzed using (SEM) electron microscope FEI IN SPECTS Company, Philips, Poland, environmental scanning without coating with a JEOL JEM-2100 electron microscope at 100k x magnification and an acceleration voltage of 120 kV. Diano X-ray diffractometer was used for investigating XRD patterns using CuKα radiation source energized at 45 kV and a Philips X-ray diffractometer  with CuK radiation source (λ = 0.15418 nm), at a diffraction angle range of 2θ from 5 to 70° in reflection mode.Chitosan was purchased from sigma Aldrich, sodium periodate was purchased from Analytical Rasayan and starting materials were purchased from Aldrich, Acros, or Fluorochem and were utilized without purification. Reaction progress was monitored by TLC on pre-coated silica gel 60 F5 was afforded as a yellow solid (81%), m.p. 300–303 °C; 1 H NMR : 4.47 , 4.95 , 7.35–7.42 , 7.64 , 8.37–8.46 , 11.00 . C 13 NMR : 39.64, 39.80, 40.13, 40.30, 83.06, 83.82, 99.99, 117.08, 117.69, 124.12, 124.29, 135.41, 135.56, 136.12, 149.13, 149.22, 149.40, 162.81, 162.95. Analysis calculated for C20H16N6O3 (388.38): C, 61.85; H, 4.15; N, 21.64; O, 12.36. Found: C, 61.84; H, 4.14; N, 21.63.A mixture of (Z)-N’-(1-(3-aminophenyl)ethylidene)-2-cyanoacetohydrazide (0.01 mol) and (Z)-ethyl 2-cyano-3-(pyridin-3-yl)acrylate (0.01 mol), was refluxed for 3 h in methanol / acetic acid (1:2). The precipitated solid product was filtered off and recrystallized from methanol. Compound 4) was added at acidic pH and the reaction container was covered with aluminium foil to avoid photo-induced decomposition of the periodate. The reaction was stopped after 24 h by adding 30 ml of ethylene glycol. The resulting dialdehyde chitosan (DACs) was washed with ethanol and dried at room temperature oxadiazine-7-carboxylic acid (5) is reported. The synthesis of the targeted structure was achieved starting from synthesizing the (Z)-N’-(1-(3-aminophenyl)ethylidene)-2-cyanoacetohydrazide (1) as a starting material by reacting the cyano acid hydrazide with the 3-amino acetophenone at reflux for hour in the presence of ethanol. The further reaction of compound 1 with the (Z)-ethyl 2-cyano-3-(pyridin-3-yl)acrylate (2) afforded the substituted PO , the peaks at 3500 cm− 1, 2200 cm− 1, 1618 cm− 1 corresponding to OH/ NH, CN cyano, C = N groups, respectively. While in PODACs (d), the peak of CN cyano is completely disappeared. The peaks of OH/NH, C = N and C-N due to the amino hydroxyl, amide and bond ether linkage at 3500 cm− 1, 1618 cm− 1, 1045 cm− 1, respectively oxadiazine (A) has improved by adding dialdehyde chitosan as in the compound PODACs (B) and it appears in the bond length (Ao) and Energy (e). where the bond length (Ao) decreased PODACs (B) than the bond length (Ao) of PO (A), which means the ligand (B) is very close to the protein pocket of bacteria and can destroy it. So it increases the biological activity oxadiazine exhibited weak antibacterial activity against E. coli and S. aureus only among all tested bacterial strains, where inhibition zones were 14 and 15 mm respectively. On the other hand, pyridooxadiazine did not exhibit any activity neither B. subtils nor C. albicans. Previous studies confirmed that ][b]oxadiazine as a try to increase antimicrobial activity. Results illustrated that, addition of dialdehyde chitosan to pyridooxadiazine led to significant increase in its antibacterial and antifungal activity against all tested bacterial and fungal strains. This composite  exhibited promising antibacterial activity against Gram-negative bacteria (E. coli) and Gram-positive bacterial (S. aureus and B. subtilis) where inhibition zones were 19, 18 and 36 mm respectively. Likewise, MIC of the composite was in accordance with inhibition zone, where MICs were 125, 125 and 15.62 µg/ml toward E. coli, S. aureus respectively  where AmB did not exhibit any activity. Therefore, the prepared composite in the current study can be used as a new antimicrobial agent in biomedical field [Antimicrobial activity of the prepared composite and compound 5 was evaluated against n Fig. b],3,4oxadied that ], 30. Then Fig. b],3,4oxadial field .50 of the prepared composite  and compound 5  toward Vero normal cell line was 472 and 419 µg/ml. Also, result showed that viability of Vero cells at different concentrations of composite 250, 125, 62.5 and 31.25 µg/mL was more than 99% for each. In general, if the IC50 is ≥ 90 µg/mL, the material is classified as non-cytotoxic oxadiazine) and composite  at concentrations ranged from 1000 to 31.25 µg/mL was carried out against cancerous cell lines MCF7 as shown in Fig. b]oxadiazine without dialdehyde chitosan exhibited weak anticancer activity where IC50 was 405 µg/ml, but pyridooxadiazine-dialdehyde chitosan exhibited anticancer activity against MCF7 higher than pyridooxadiazine only where IC50 was 238 µg/ml as shown in Table Anticancer activity of compound 5  through reacting(Z)-N’-(1-(3-aminophenyl)ethylidene)-2-cyanoacetohydrazide with (Z)-ethyl 2-cyano-3-(pyridin-3-yl)acrylate. Then, a novel composite has synthesised based on substituted pyridooxadiazine-dialdehyde chitosan conjugate (PODACs). The prepared compounds have investigated via FTIR, XRD, and SEM. The prepared composite exhibited promising antimicrobial activity against E. coli, S. aureus, B. subtilis and C. albicans. Furthermore, the composite illustrated cytotoxic activity against MCF-7 human breast cancer cell line as well as Vero normal cell line.This work reported dialdehyde chitosan (DACs) preparation via periodat oxidation of chitosan and preparation of a new substituted pyrido[2,1-Below is the link to the electronic supplementary material.Supplementary Material 1


Scheffersomyeces spartinae W9 is a promising biocontrol agent for gray mold caused by Botrytis cinerea in strawberries. Improving the biocontrol efficacy of S. spartinae W9 is necessary for its commercial application. In this study, different concentrations of β-glucan were added to the culture medium to evaluate its effect on the biocontrol efficacy of S. spartinae W9. The results showed that 0.1% β-glucan could increase the biocontrol effect of S. spartinae W9 against B. cinerea in strawberries and in vitro. We found that adding 0.1% β-glucan to the culture medium promoted the growth of S. spartinae W9 in wounds of strawberries, enhanced biofilm formation ability, and secreted more β-1,3-glucanase. In addition, 0.1% β-glucan increased the survival rate of S. spartinae W9 under oxidative, thermal, osmotic, and plasma membrane stressors. Transcriptome analysis revealed 188 differential expressed genes in S. spartinae W9 cultured with or without 0.1% β-glucan, including 120 upregulated and 68 downregulated genes. The upregulated genes were associated with stress response, cell wall formation, energy production, growth, and reproduction. Thus, culturing with 0.1% β-glucan is an effective way to improve the biocontrol ability of S. spartinae W9 against gray mold in strawberries.The marine yeast  Botrytis cinerea is the primary fungal disease of strawberry fruit during planting and postharvest storage . A S. spartinae W9 suspensions  at a concentration of 1 × 105 cells/mL was spread into the PDA medium. A total of 100 µL of sterile water was spread to the PDA and used as the control. Afterwards, a 9-mm diameter plug taken from the edge of the actively growing B. cinerea colony was inoculated into the center of each PDA plate. The plates were incubated at 28 °C for 5 d, and the mycelia diameters were measured daily. The experiment consisted of three replicates, with 5 plates per replicate. For the mycelia growth test, 100 µL of 9 cells/mL S. spartinae W9  were prepared. Then 200 µL of yeast suspension was mixed with 200 µL of B. cinerea (1 × 106 spores/mL) spore suspension and added into a 1.6-µL PDB medium. An amount of 200 µL sterile water mixed with 200 µL B. cinerea spore suspension was used as the control. The spores were incubated for 6 h at 28 °C and 180 rpm on a shaker. The germination rate of B. cinerea spores were counted by light microscopy and hemocytometer. At least 100 spores were observed in each treatment, with 3 replicates per treatment.For the spore germination test, cell suspensions of 1 × 10S. spartinae W9 or 0.1% β-glucan-treated S. spartinae W9. All strawberries were stored at 20 °C with 90% relative humidity. The amount of yeast at the strawberry fruit wounds was measured at 0 h, 12 h, 24 h, 48 h, and 72 h. The fruit wound tissue was removed with a sterile blade and homogenized in a mortar containing 10 mL of sterile water. A total of 100 µL of the homogenate diluted to the appropriate concentration was spread on NYDA and incubated at 28 °C for 48 h to count the number of colonies. The number of yeast colonies at each wound was expressed as log10CFU/wound. The experiment consisted of three replicates, with three strawberries per replicate.In accordance with the method of Zou et al. (2022) , strawbeS. spartinae W9 cells (1 × 108 cells/mL) were cultured in NYDB or NYDB with 0.1% β-glucan at 28 °C. The growth of S. spartinae W9 was recorded by measuring the absorbance at 600 nm at 12 h, 24 h, 36 h, and 48 h. Three biological replicates were available for each treatment.S. spartinae W9 was assessed using the method of Qiu et al. (2022)  wS. spartinae W9 to produce extracellular hydrolases, mainly including chitinase and β-1,3-glucanase (GLU), was determined by an extracellular hydrolase assay plate [S. spartinae W9 secreted chitinase and CLU, the hyaline rings would appear in the plates.The ability of ay plate . ChitinaS. spartinae W9. S. spartinae W9 cultured in NYDB or NYDB with 0.1% β-glucan were sampled at 12, 16, 20, 24, 36, and 48 h. The fermentation broth was centrifuged at 6000 rpm for 10 min, and the supernatant was filtered through a sterile membrane for enzyme assay. The GLU activity was determined using the 3, 5-dinitrosalicylic acid method [According to the result of an extracellular hydrolase assay plate, we determined the GLU activity of d method , and the2O2, and sodium dodecyl sulfate (SDS) were used to simulate stress. S. spartinae W9 were cultured in NYDB or NYDB with 0.1% β-glucan for 24 h, and the yeast suspensions (1 × 108 cells/mL) were prepared. As described by Huang et al. (2021) , high teS. spartinae W9 to high temperatures, the yeast suspensions were placed into a water bath at 40 °C, 45 °C, and 50 °C for 30 min. The heat-treated yeast suspensions were cooled at room temperature for 10 min and diluted to the appropriate concentration. Then, 0.1 mL of each yeast suspension was spread on a NYDA plate to calculate the survival rate. To determine the tolerance of S. spartinae W9 or β-glucan-treated S. spartinae W9 yeast suspensions were injected into NYDB (control) and a selective medium . Yeasts were incubated at 28 °C and 180 rpm for 24 h and then diluted to the appropriate concentration to spread on NYDA plates. All plates were incubated at 28 °C for 48 h, and the numbers of yeast colony were recorded. The survival rates of S. spartinae W9 or β-glucan-treated S. spartinae W9 without stress treatment were used as respective controls. Each treatment contained three independent replicates.S. spartinae W9 cultured in NYDB with or without 0.1% β-glucan was isolated using TRIzol  according to the manufacturer’s instructions. The mRNA with PolyA (polyadenylation) was specifically captured and fragmented with a magnesium ion interruption kit . The fragmented RNA was synthesized into cDNA. Then, double-stranded DNA were synthesized and repaired. The fragment sizes were screened and purified using magnetic beads, and the cDNA library was enriched with PCR. RNA-seq was performed by LC-Bio Technology CO., Ltd. Hangzhou, China. Genes with a fold change in expression level ≥ 1.2 and adjusted p < 0.01 were identified as differentially expressed genes (DEGs). The DEGs’ data was analyzed for GO and KEGG enrichment analysis. RNA from S. spartinae W9 cultured in NYDB with or without 0.1% β-glucan were first reverse transcribed to cDNA. RT-qPCR was performed using ChamQ Universal SYBR qPCR Master Mix . Twelve DEGs were randomly selected for validation by qPCR, and ACT1 was used as an internal reference gene. Relative gene expression was calculated according to the 2−ΔΔCt method. RNA extracted from t-test. Significance was assessed when p < 0.05. Data were expressed as mean ± standard deviation.SPSS statistics software version 20 was used to analyze the data. When the number of comparisons within the group was three or more, the data were compared with the mean by implementing Duncan’s multiple analysis using ANOVA; when the number of comparisons within the group was two, the mean was compared using the independent samples S. spartinae W9 cultured with or without β-glucan reduced the disease incidence of strawberries. S. spartinae W9 cultured with 0.1% β-glucan significantly reduced the disease incidence when compared to yeast without β-glucan after the fourth day (p < 0.05). At the end of storage, the disease incidence was 96.7% in the control group, 63.3% in the S. spartinae W9 group, and only 40% in the 0.1% β-glucan-treated S. spartinae W9 group . Compared to the S. spartinae W9 cultured without β-glucan, 0.1% and 1% β-glucan were effective in increasing the inhibition of B. cinerea mycelial growth, for which the diameter of B. cinerea plaques in the S. spartinae W9 treatment group was 23.4 mm, while the diameters of the 0.1% and 1% β-glucan-induced S. spartinae W9 treated groups were 18.8 mm and 18 mm, respectively. As shown in S. spartinae W9 cultured with or without β-glucan significantly inhibited spore germination of B. cinerea (p < 0.05). S. spartinae W9 cultured with different concentrations of glucan showed different inhibitory effects on spore germination of B. cinerea. The germination rate of B. cinerea spores in 0.1% β-glucan-treated S. spartinae W9 was 9.5%, which was significantly lower than the 17.1% in the S. spartinae W9 group (p < 0.05). Only 0.1% β-glucan increased the ability of S. spartinae W9 to inhibit spore germination of B. cinerea.S. spartinae W9 yeasts were cultured in NYDB and NYDB containing 0.1% β-glucan at a concentration of 0.1% for 48 h, respectively. The OD600 of S. spartinae W9 in both media showed no significant difference . The growth rate of the β-glucan-treated S. spartinae W9 within 24 h was higher than S. spartinae W9 cultured in NYDB. at 24 h A. The poS. spartinae W9. During the 24 h of yeast culture, the biofilm formation capacity remained stable in each group , which showed that higher GLU activity of C. podzolicus is an important factor for the increased biocontrol effect of C. podzolicus cultured with β-glucan [The ability of antagonistic yeast to resist pathogens also depends on the secretion of extracellular hydrolase. GLU can hydrolyze glucan, which is the major component of fungal cell walls, leading to cell malformation and cytoplasmic leakage, and is one of the possible mechanisms of biocontrol of antagonistic yeasts . GLU is tivities . This suβ-glucan ,18.S. spartinae W9 was significantly inhibited under these stressors, while S. spartinae W9 cultured with 0.1% β-glucan could enhance tolerance under these stressors  showed that inactivation of ADH1 caused the accumulation of NADH, resulting in the imbalance of NADH/NAD and growth retardation [ACH1 and ADH1 in S. spartinae W9 cells cultured with 0.1% β-glucan led to higher colonization ability in strawberry wounds and improved biocontrol efficacy. In addition, upregulation of the RNR2 gene required for DNA repair may be helpful for maintaining cellular integrity [S. spartinae W9 cells.Intracellular energy is heavily depleted when yeast cells are under abiotic stress . ACH1 prardation . The uprntegrity . In brieS. spartinae W9 achieved better proliferation and enhanced biological activity by altering cellular processes, energy metabolism, and stress responses. Similarly, Zhao et al. (2020) used transcriptomic analysis to reveal that β-glucan accelerated the synthesis of cell walls and energy of C. podzolicus, enhanced its antioxidant ability, and thus improved the biological control efficacy [The transcriptome analysis demonstrated that the 0.1 % β-glucan-induced efficacy .S. spartinae W9 against B. cinerea in strawberries. β-glucan improved the colonization ability of yeast in strawberries, the biofilm forming ability, and the stress tolerance of S. spartinae W9. β-glucan also upregulated the genes involved in stress responses, cell wall formation, energy metabolism, growth, and reproduction of S. spartinae W9.Adding 0.1% β-glucan into the culture medium significantly enhanced the biocontrol efficacy of


Alzheimer’s disease (AD) is the most common cause of dementia in elderly people; currently, there is no efficient treatment. Considering the increase in life expectancy worldwide AD rates are predicted to increase enormously, and thus the search for new AD drugs is urgently needed. A great amount of experimental and clinical evidence indicated that AD is a complex disorder characterized by widespread neurodegeneration of the CNS, with major involvement of the cholinergic system, causing progressive cognitive decline and dementia. The current treatment, based on the cholinergic hypothesis, is only symptomatic and mainly involves the restoration of acetylcholine (ACh) levels through the inhibition of acetylcholinesterase (AChE). Since the introduction of the Amaryllidaceae alkaloid galanthamine as an antidementia drug in 2001, alkaloids have been one of the most attractive groups for searching for new AD drugs. The present review aims to comprehensively summarize alkaloids of various origins as multi-target compounds for AD. From this point of view, the most promising compounds seem to be the β-carboline alkaloid harmine and several isoquinoline alkaloids since they can simultaneously inhibit several key enzymes of AD’s pathophysiology. However, this topic remains open for further research on detailed mechanisms of action and the synthesis of potentially better semi-synthetic analogues. The World Alzheimer Report 2019 pointed out that with the continuous acceleration of the aging population, the number of AD patients worldwide had exceeded 50 million, but the number of patients is expected to double by 2030 and to reach 152 million by 2050. Moreover, AD has a significant influence on health and economic development that needs to be addressed urgently .,50.49,50In June 2021, a new AD drug named aducanumab, sold under the brand name Aduhelm™, was approved by the FDA in the USA for people with mild symptoms of AD, such as individuals who are still independent in basic daily functioning . AducanuAD is a complex disorder involving multiple factors that necessitate the need to identify and develop hybrid molecules that can target two or more pathological changes in the brain of AD patients . NaturalCatharanthus roseus (L.) G.Don (Apocynaceae); and taxanes, isolated from the genus Taxus L. (Taxaceae), which are used as chemotherapy agents indole-7-carboxylic acid methyl ester; 50 values of 71 ± 9 nM and 103 ± 4 nM, respectively .Suzuki coupling and Cadogan cyclization were used for the preparation of HAR and a series of fifteen derivatives, which were screened for dual GSK-3β/DYRK1A inhibitory activity. Among them, the carbonyl group as a hydrogen bond acceptor containing compound ZDWX-25 -9ectively . MoleculBerberine (BBR) and palmatine (PAL) are quaternary isoquinoline alkaloids of protoberberine-type, biosynthetically derived from tyrosine . Both alCoptis chinensis Franch. (Ranunculaceae), Berberis vulgaris L. (Berberidaceae), Hydrastis canadensis L. (Ranunculaceae), and Phellodendron amurense Rupr. (Rutaceae). For decades, Chinese medicine has used plants and their extracts containing BBR to treat various diseases [The Divine Farmer’s Herb-Root Classic [BBR is commonly found and isolated from the roots, rhizomes, stems, and bark of diseases . BBR was Classic . BBR sho Classic . For the Classic . Moreove Classic ,105,106; Classic .42 production, AChE activity, and inflammation [50 for BACE-1 was in another study determined to be higher than 100 μM [40/42 production by BBR was also confirmed by a study on HEK293 cells, which can be explained by the inhibition of the expression of BACE via activation of the extracellular signal-regulated kinase 1/2 pathway [50 for the inhibition of Aβ1–40 of 43.84 μM (50 for the induction of Aβ1–40 disaggregation (104.90 μM) [hAChE, its IC50 ranges from 0.52 μM to 0.7 μM with Ki = 0.54 μM [50 values are several times higher, being up to 30.7 μM for hBuChE inhibition [50 of 126 μM [In recent years, BBR has been extensively investigated by various researchers for its activity against AD; numerous studies have indicated that BBR treatment significantly improves memory and cognitive dysfunction in different animal models of AD ,107. Forammation . In the ammation . Anotherammation . BBR in ammation . Using tammation . BBR’s cammation . In a raammation ; howevern 100 μM . BBR hasn 100 μM . The dec pathway . The stu43.84 μM , as well4.90 μM) . Additio4.90 μM)  and by i4.90 μM) , further4.90 μM) . In HEK24.90 μM) . Simulta4.90 μM) , as well4.90 μM) . It is n 0.54 μM ,83. Compf 126 μM ; howeverf 126 μM . BBR hasf 126 μM . Lastly,f 126 μM .Coptis chinensis, and Corydalis DC. species (Papaveraceae), and many other herbs used in traditional Chinese medicine [50 values ranging from 0.46 μM [hAChE [50 for hBuChE > 100 μM [eqBuChE; 50 = 47 μM [C. elegans AD model containing human Aβ1–42, showing that treatment significantly delayed the paralytic process, reduced the amount of oxidative stress, and alleviated the deposition of Aβ [PAL can be found in the roots of medicine ,126,127. 0.46 μM  to 1.69 M [hAChE ; however> 100 μM , and 6.8 = 47 μM . PAL was = 47 μM . Further = 47 μM . The neuon of Aβ . The capon of Aβ .hAChE in vitro, revealing the synergic action of these two protoberberine alkaloids [The study of Mak et al. elucidated the effect of simultaneous administration of BBR and PAL on the inhibition of lkaloids . The higlkaloids . PAL andlkaloids . OverallZanthoxylum rigidum Humb. et Bonpl. ex Willd. (Rutaceae) for detailed phytochemical study [EeAChE, hAChE, and eqBuChE inhibitory activity, as well as for MAO-A and B inhibition, and Aβ aggregation. Avicine contains in its structure a dioxomethylene bridge connecting positions 8 and 9, while in nitidine, these positions are substituted by two methoxy groups micromolar concentration range , with an eqBuChE/hAChE selectivity index of 1.67. Kinetic studies indicated that avicine and nitidine are reversible-mixed inhibitors of both cholinesterases. Mixed-type inhibitors are able to bind at the catalytic and peripheral anionic sites (PAS) of the enzyme. PAS inhibitors can regulate ChE-induced Aβ aggregation, which supported results obtained within the study, since avicine and nitidine were able to inhibit Aβ1–42 aggregation with IC50 values of 5.56 ±0.94 µM and 1.89 ± 0.40 µM, respectively [50 > 100 µM).Preliminary screening studies led to the selection of al study . Multi-sy groups . Both coon range . Both alectively . In the Chelidonium majus L. (Papaveraceae) [hAChE (1.54 ± 0.07 µM) than towards the EeAChE (3.78 ± 0.15 µM). A reversed situation was obtained for BuChE, as chelerythrine was slightly more active towards the horse eqBuChE (6.33 ± 0.95 µM) when compared to the human enzyme . The same mechanism of inhibition was found using hAChE with a icK = 0.32 ± 0.08 µM and icK = 1.12 ± 0.07 µM. Based on these results, a docking study of chelerythrine was performed using crystal structure PDB ID: 1FSS  as a model receptor, as this structure is fundamentally similar to that of human AChE [1–40 aggregation [1–40 aggregation was studied using 1,1,1,3,3,3-hexafluoro-2-propanol as an aggregation enhancer [1–40 aggregation at a concentration of 10 µM; thus, the value of IC50 was subsequently determined (4.2 ± 0.43 µM), which indicated that chelerythrine is a highly active inhibitor of Aβ1–40 aggregation. Moreover, chelerythrine was also tested for its ability to disaggregate already preformed Aβ1–40 aggregates. This ability could be more relevant from a clinical perspective, when disaggregation of already existing Aβ fibrils in the AD brain could be indicated, especially at the beginning of the treatment to reduce the neurotoxic effects of Aβ fibrils and thus prevent neurodegeneration. Chelerythrine showed a high activity in disaggregating preformed Aβ1–40 aggregates, with an IC50 of 13.03 ± 2.89 µM after 45 min incubation [A further benzophenanthridine alkaloid chelerythrine, commonly isolated from eraceae) , containman AChE . Cheleryman AChE . Cheleryregation . The infenhancer . Chelerycubation .50 value of 0.55 ± 0.042 µM [iK = 0.22 ± 0.033 µM). Docking simulation showed that chelerythrine binds to MAO-A due to two hydrogen bond interactions with Cys323 and Tyr444 [Chelerythrine also selectively inhibited an isoform of recombinant human MAO-A with an IC0.042 µM  and was d Tyr444 .Almost all of the current natural product-derived therapeutics have terrestrial origins. However, there are four approved marine or marine-derived drugs , which are used as either anti-viral or anticancer agents  136,137137. ThusParazoanthus axinellae, exhibited anticholinesterase activity. Subsequent RP-HPLC separation led to the isolation of pseudozoanthoxanthin (PSX), containing 2-amino imidazole groups in its structure  theta rhythm during a cognitive performance in an experimental model of AD [An ethanolic extract of a zoanthid crust coral, tructure , which hi = 4 µM . Whereasi = 4 µM , this ali = 4 µM . For the eqBuChE . Docking0 values  and werepeptides . Moreoveomplexes . Furtherescribed . These eel of AD ,148. PSXAelasidae, Axinellidae, and Halichondriade families, has been identified as an inhibitor of various kinases within marine alkaloids carrying an imidazole-core in the structure [50 > 1 µM); however, four, CDK1/cyclin B, CDK5/p35, GSK-3β, and casein kinase 1 (CK1), were strongly sensitive to HD  of 20 µM. In a follow-up study, PLMB showed balanced inhibitory activity against both cholinesterases with IC50 values of 37.02 ± 2.11 µM for AChE and 30.70 ± 1.44 µM for BuChE [1–42 aggregation using the ThT fluorescence method. Tacrine and donepezil were used as positive control. PLMB is a weaker inhibitor of self-induced Aβ1–42 aggregation (29.78 ± 1.45% at 10 µM) compared to donepezil. On the other hand, PLMB showed comparable potency to inhibit AChE-induced Aβ1–42 aggregation (27.60 ± 1.96% at 10 µM) as donepezil (22.42 ± 2.56% at 10 µM) [50 > 80 µM).Pulmonarin B (PLMB), a dibrominated phenylacetic acid derivative containing a quaternary ammonium group, has been isolated from the ascidian  potency . In thisor BuChE . PLMB wat 10 µM) . MoreoveAspergillus ochraceus, together with other circumdatins [50 value of 2.4 ± 0.5 µM, and thus it was selected for further experiments. In vitro, CIRD has demonstrated neuroprotective effects by inhibiting the toll-like receptor 4-mediated NF-κB, mitogen-activated protein kinases, and Janus kinase/signal transducer and activator of transcription protein signaling pathways responsible for neuroinflammation. Furthermore, this alkaloid has been shown to substantially inhibit the lipopolysaccharide (LPS)-induced production of NO and cytokines such as TNF-α, IL-1β, and cyclooxygenase-2 expression in microglial BV-2 cells. CIRD-treated neuronal cells have also been observed to have significantly reduced LPS-induced AChE activity. The AD model of C. elegans strain CL4176 was used for in vivo assay of CIRD. This model expresses Aβ in muscle cells, which leads to progressive neurodegeneration and paralysis [C. elegans [Circumdatin D CIRD,  is a quiumdatins , and subumdatins  or gorgoumdatins . This coaralysis . Moreove elegans .Multi-factorial diseases like AD require complex treatment strategies that involve simultaneous modulation of a network of interacting targets. During the last few years, the multi-target compounds have been explored as an effective therapeutic approach for the treatment of AD. Besides direct inhibition of enzymes involved in AD pathology, intensively studied targets include amyloid plaque deposition, neuroinflammation signaling pathways, anti-apoptotic and anti-oxidative stress activities, and neuroprotection.In the current review article, which is focused on alkaloids and other nitrogen compounds, various pathogenic pathways associated with AD have been briefly described, followed by an overview of selected natural alkaloids that can be recognized as multi-target compounds for the development of new anti-AD drugs. Of these, harmine is the most promising alkaloid, displaying a wide spectrum of compelling anti-AD activities. Isoquinoline alkaloids such as berberine, avicine, and chelerythrine also appear to be promising multi-target compounds, exhibiting strong inhibitory activity on key pathological enzymes of AD. Furthermore, marine flora have emerged as a viable source of multi-target compounds as well; for example, hymenialdisine has a broad range of protein kinase-inhibiting activities in a nanomolar range. Such compounds are currently of interest for the preparation of semi-synthetic derivatives.To conclude, it is essential to undertake more studies on the discussed alkaloids at a cellular and molecular level, such as influence on the various signal pathways connected to the neurodegeneration, docking studies and molecular dynamics studies in the active sites of the target enzymes, and cytotoxicity evaluations to select clinically important candidates for treating neurodegenerative diseases.