Active Fraction Research Articles

Evaluation of Anti-angiogenic Activity of Iraqi Mesembryanthemum cordifolium using rat aorta ring assay

New blood vessels are produced during angiogenesis, which is necessary in both normal and abnormal physiological and pathologic states. The use of non-toxic plant extracts that inhibit angiogenesis might lead to more focus on the use of natural sources as a pivotal therapeutic agent in the treatment of many diseases and malignancies. Mesembryanthemum cordifolium is belongs to the Aizoaceae family and commonly known as the Heartleaf Ice Plant or Tiny Sun Rose. The plant was cultivated in the city of al Diwaniyah. No previous research has been conducted on this plant and its antiangiogenic properties. Hence, this research aimed to assess the antiangiogenic capacity of the Iraqi M. cordifolium. For this study, male albino rats aged 12 to 14 weeks were used. With a Soxhlet apparatus, the defatted plant components were extracted in 85 % methanol for 12 h. Subsequently, each fraction was fractionated using petroleum ether, chloroform and ethyl acetate and each fraction underwent phytochemical analysis. Rat aortic rings were infused with 100 ?g/mL of each plant component and placed in a growth medium. In addition, the dose-response relationship was assessed for the most active fraction at 6 different concentrations ranging from 6.25 ?g/mL to 100 ?g/mL. Compared to the negative control (1 % DMSO), the results demonstrated that fractions of M. cordifolium inhibit the growth of blood vessels in a dose-dependent manner; chloroform demonstrates the most potent effect. The results indicate that the chloroform fraction derived from M. cordifolium has a good ability to limit blood vessel development. This makes it a viable option for future investigation as an angiogenesis inhibitor.

Value of 2D speckle tracking technique combined with real-time 3-dimensional echocardiography in the evaluation of the right atrial function in patients with 3-branch coronary artery disease without myocardial infarction.

To evaluate the right atrial function in patients with 3-branch coronary artery disease (TBCAD) without myocardial infarction by 2D speckle tracking echocardiography (2D-STE) combined with real-time 3-dimensional echocardiography (RT-3DE). Fifty-six patients admitted to our hospital without myocardial infarction with TBCAD were selected. We divided them into 2 groups according to the coronary angiography results: 28 patients in group B (the rate of stenosis is 50% ~< 75%); 28 patients in group C (the rate of stenosis is ≥75%); in addition, 30 healthy volunteers were screened as group A. All subjects underwent RT-3DE to obtain the right atrial volume (RAVmax, RAVmin, and RAVp), and then we calculated the right atrial passive and active ejection fraction (RAPEF, RAAEF), and maximum volume index (RAVImax). In addition, to measure the strain rates (RASRs, RASRe, RASRa) of the right atrium during systole, early diastole, and late diastole, 2D-STE was applied. Correlations between the 2D-STE parameters and the results of N-terminal pro-brain natriuretic peptide (NT-proBNP) and Gensini scores were analyzed by Pearson linear analysis. Compared with group A, RAPEF and RASRe were reduced, while RAAEF and RASRa were elevated in group B (P < .05). RAPEF, RASRs, RASRe, and RASRa were decreased compared with groups A and B, while RAVmax, RAVmin, RAVp, RAVImax, and RAAEF were increased in group C (P < .05). There was a significant correlation between 2D-STE parameters and the results of NT-proBNP and Gensini scores (P < .05). The storage, conduit, and pump functions of the right atrium are reduced in patients with 3-branch coronary artery disease without myocardial infarction; 2D-STE combined with RT-3DE is valuable in the evaluation of the right atrium in patients with coronary artery disease.

Investigating the Anti-adipogenic Potential of Xanthium strumarium Linn. Leaves Fractions: HPTLC-MS Characterization, Enzymatic Analysis, and 3T3-L1 Adipocyte Differentiation Assay in Oxidative Stress-modulated Adipogenesis Cascade

Background: Molecular pathogenesis of obesity is initiated by cellular oxidative stress along with increased adipogenesis. For the search of better alternatives of synthetic molecules in terms of their adverse events, naturally, existing compounds are now a research focus. In this context, Xanthium strumarium ( X. strumarium) Linn. exhibited anti-diabetic activities and proved to have efficacy in neutralizing very near pathological targets to obesity. Purpose: This study is aimed toward bioactivity-guided fraction isolation of methanolic leaves extract, where bioactivity refers to anti-adipogenesis and anti-oxidant efficacy in in vitro enzymatic and three-day transfer, inoculum 3×105 cells (3T3-L1) adipocyte differentiation assay. Materials and method: Collection and drying of the raw leaves followed by cold extraction in methanol and flash chromatography for bioactive fraction isolation. 3T3-L1 cell line was utilized for adipocyte differentiation assay of those and on the other hand in vitro pancreatic lipase, α-glucosidase, and α-amylase enzymatic assays were analyzed. Based on exhibited activities, selected fractions are further characterized by high-performance thin-layer chromatography-mass spectrometry (HPTLC-MS). Results: The most active fractions from these assays underwent characterization via HPTLC-MS. Fractions 2 and 3 exhibited potent activities in all enzymatic assays (α-glucosidase IC50: 2.48 ± 0.015, 2.84 ± 0.030 µg/mL; α-amylase IC50: 1.98 ± 0.050, 1.79 ± 0.045 µg/mL; pancreatic lipase IC50: 3.16 ± 0.030, 3.18 ± 0.040 µg/mL) and displayed significant inhibition of adipocyte differentiation in 3T3-L1 cell line. These fractions further identified through HPTLC-MS, contained compounds like β-sitosterol and quercetin, affirming their potential bioactivity. Conclusion: The study underscores the potential of X. strumarium Linn. fractions as promising natural alternatives for combating obesity-related pathways, highlighting their significance in developing future anti-obesity therapeutics.

Chinese medicine Di-Long (Pheretima vulgaris) and its active fraction exhibit anti-rheumatoid arthritis effects by inhibiting CXCL10/CXCR3 chemotaxis in synovium

Ethnopharmacological relevanceDi-Long (Pheretima vulgaris) is a classic animal sourced traditional Chinese medicine. It has been used for the treatment of joint inflammation and arthralgia for over two thousand years due to its effects of Tong-Luo-Zhi-Tong (dredging collaterals and alleviating pain). Our previous study showed that Chinese medicine Di-Long has significant anti-rheumatoid arthritis (RA) effects. Aim of the studyConsidering Di-Long as a potential source of active compounds with specific anti-RA therapeutic effects, this research was to obtain the anti-RA target-specific active fraction from Di-Long extracts (DL), and to further explore the chemical basis and verify the anti-RA mechanism of this active fraction. Materials and methodsTranscriptomic was applied to obtain the main anti-RA targets of DL on human RA fibroblast-like synoviocytes (FLS) and validated by qPCR. The target-corresponding active fraction was isolated from DL by ethanol precipitation and gel chromatography, and analyzed by nanoliter chromatography-mass spectrometry. Anti-RA effects of this active fraction was investigated by collagen-induced arthritis (CIA) in mice, and anti-RA mechanisms were verified in cocultured model of rat FLS and peripheral blood lymphocytes. ResultsWe confirmed that CXCL10/CXCR3 was the main anti-RA target of DL. The active fraction - A (2182 - 890 Da) was isolated from DL based on its CXCL10 inhibiting effects in RA-FLS. Fraction A contains 195 peptides (192 were newly discovered), 26 of which might be bioactive and were considered to be the chemical basis of its anti-RA effects. Fraction A significantly ameliorated the joint destruction and overall inflammation in CIA mice, and downregulated CXCR3 expression in mice joint. Fraction A inhibited the chemotaxis of Th-cells in rat peripheral blood lymphocytes towards the TNF-α-induced rat FLS through CXCL10/CXCR3 pathway. ConclusionsOur work indicated that active fraction from DL containing small peptides exhibits promising therapeutic effects for RA through inhibiting CXCL10/CXCR3 chemotaxis.

Fractionation and identification of bioactive peptides from red macroalgae protein hydrolysates: in silico analysis and in vitro bioactivities

Recently, bioactive peptides derived from algae sources have gained significant recognition as vital functional ingredients in the development of health-enhancing foods and nutraceuticals. In this study, protein hydrolysates (SCPH and GSPH) derived from two red macroalgae (Sphaerococcus coronopifolius and Gelidium spinosum) were fractionated using semi-preparative-reversed phase high-performance liquid chromatography (RP-HPLC). Then, the peptides contained in the obtained fractions were identified by RP-HPLC coupled with tandem mass spectrometry (MS/MS). In silico analyzes of hydrophobicity, potential bioactivity using PeptideRanker, and toxicity of identified peptides were performed. Moreover, the prediction of Angiotensin-converting enzyme (ACE)- and Dipeptidyl-peptidase (DPP-IV)-inhibitory peptides using a quantitative structure-activity relationship (QSAR) was also carried out. Furthermore, in vitro evaluation of antioxidant and antibacterial activities, as well as ACE-, and DPP-IV-inhibitory properties was conducted for all peptide fractions. Our results showed that SCPH and GSPH fractions contain a large percentage of highly hydrophobic peptides with low molecular weight, and they exhibit no toxic effect. Among all identified peptides, 61 and 39 peptides, respectively from SCPH and GSPH fractions, presented a PeptideRanker score superior to 0.8. These peptide sequences exhibited a high proportion of potent predicted ACE- and DPP-IV-inhibitory peptides. Additionally, most of the peptide fractions exerted antioxidant, antibacterial, anti-ACE, and anti-DPP-IV inhibitory activities at different levels. Among them, F2 to F6 were the most active fractions. Overall, the results suggest that the bioactive peptide fractions separated from two red macroalgae protein hydrolysates could be considered as a beneficial ingredient for nutraceutical and functional food industries.

CERVUS ELAPHUS: CULTIVATION IN THE KALININGRAD REGION, PROCESSING AND EVALUATION OF THE QUALITY AND SAFETY OF VENISON PRODUCTS

Recently, there has appeared a tendency to use non-traditional animal raw materials to expand the range of sausages. The purpose of the research is to determine the organoleptic and physico-chemical properties, as well as quality indicators of new sausages with a mass fraction of deer muscle tissue in the recipe (&gt; 60%). An organoleptic evaluation was carried out, then the study of physicochemical parameters, the mass fraction of protein, fat, salt, sodium nitrite, total phosphorus, and residual activity of acid phosphatase was performed. Next there were studied such microbiological indicators as QMAFAnM, BGKP (coliforms), sulfite-reducing clostridia, Staphylococcus aureus, and pathogenic microorganisms: Salmonella, Escherichia coli, Listeria monocytogenes. Safety indicators have been established: the content of toxic elements (mercury, arsenic, cadmium, lead) and quality indicators: the content of nitrosamines, pesticides. The quality control of packaging and labeling was carried out visually in accordance with the requirements of the standard. The temperature is measured inside the product with a glass liquid thermometer. The net weight of consumer packaging was determined gravimetrically by static weighing. It has been proved that all three types of the studied sausages «Zavtrak okhotnika», «Drezdenskiye» and «Oldenburgskiye» corresponded to the normative documentation in terms of organoleptic, physico-chemical properties, quality and safety indicators, nutritional value.

Rapid screening of potential α-glucosidase inhibitors from the waste leaves of Rheum tanguticum by activity-oriented extraction and enrichment optimization, UPLC-QTOF-MS/MS, molecular docking and in vitro validation

In the present study, an efficient strategy has been established for rapid screening of potential α-glucosidase inhibitors from the waste leaves of Rheum tanguticum by activity-oriented extraction and enrichment optimization, UPLC-QTOF-MS/MS, molecular docking, and in vitro validation. With α-glucosidase inhibitory activity as the evaluation index, the extraction process was optimized by response surface methodology and the enrichment process by macroporous resins was optimized by adsorption and desorption experiments. The active flavonoids yield was 8.85 mg/g under the optimized extraction conditions. After AB-8 macroporous resins enrichment, the active flavonoids purity increased from 2.89 % to 47.6 %, a 17-fold increase, while the IC50 of α-glucosidase reduced from 13.97 mg/mL to 412 μg/mL, a 33.9-fold reduction. Thirty-six flavonoids in the active flavonoids fraction were identified by UPLC-QTOF-MS/MS. Molecular docking screening, in vitro activity assay and multispectral approaches verification confirmed the potential of six flavonoids as novel α-glucosidase inhibitors.

Ultrasonication assisted percolation extraction, isolation, identification and in silico studies of secondary metabolites from Caesalpinia bonduc (L.) Roxb. seeds with special reference to the target proteins involved in Benign Prostatic Hyperplasia

Medicinal plant-derived phytochemicals provide a vast pool of diverse bioactive compounds for novel drug development. Caesalpinia bonduc L. Roxb. is a traditional medicinal plant that is reported for diverse pharmacological properties. This study aims to compare conventional (maceration) and novel extraction techniques (ultrasonication assisted maceration extraction) to evaluate their efficiency, yield and bioactive potential of hydroalcoholic extracts from C. bonduc seeds. Furthermore, the study also aimed to isolate the bioactive compounds from the hydroalcoholic seed extract obtained from ultrasonication assisted maceration and in-silico studies of isolated compounds against the protein targets involved in Benign Prostatic Hyperplasia (BPH). For comparing the influence of extraction methods, the seeds were extracted using a novel and conventional techniques. Chemical profiling, antioxidant activities, cytotoxic and apoptotic studies were conducted using obtained extracts. The extract yielded from the novel technique recorded the highest yield, significant antioxidant activity, cytotoxicity and chemical content when compared to the conventional technique. From the hydroalcoholic extract, three major bioactive compounds, namely ethyl glucoside, methyl lineolate and linoleic acid, were isolated using column chromatography. The structures of the compounds were elucidated on the basis of extensive NMR (1H, 13C, DEPT, 2D NMR- HMBC and HSQC) and HR-ESI-MS analysis. In-silico anti-BPH activity of isolated compounds clearly reveals that the compounds were targeting 5α‒Reductase and α1-Adrenoceptor linked to the BPH progressions. Based on this investigation, it is demonstrated that isolated molecules or active molecule-enriched fractions of C. bonduc should be further researched to confirm the anti-BPH activity through in-depth preclinical and clinical research for the development of products. Ethyl glucoside and methyl lineolate were isolated for the first time from C. bonduc.

Evaluation of hemostatic, anti-inflammatory, wound healing, skin irritation and allergy, and antimicrobial properties of active fraction from the ethanol extract of Chromolaena odorata (L.) R.M. King & H. Rob

Evaluation of hemostatic, anti-inflammatory, wound healing, skin irritation and allergy, and antimicrobial properties of active fraction from the ethanol extract of Chromolaena odorata (L.) R.M. King & H. Rob

Modified Zhenwu Decoction suppresses chronic colitis via targeting macrophage CCR2/Fyn/p38 MAPK signaling axis

BackgroundUlcerative colitis (UC) is associated with intestinal macrophage infiltration due to disruption of the mucosal barrier and bacterial invasion. Therefore, it is crucial to identify therapeutic agents capable of attenuating the macrophage-induced inflammatory response to preserve mucosal homeostasis and immune tolerance. The modified Zhenwu decoction (CDD-2103) is a novel herbal formulation developed based on the principles of Traditional Chinese medicine. To date, there are no clinically approved herbal formulations for UC with a well-known mechanism of action on macrophages. PurposeThe objective of this study was to systematically investigate the inhibitory effect of the active fraction of CDD-2103 in a mouse model of chronic colitis and delineate the mechanisms underlying its inhibitory action. MethodsCDD-2103 was extracted into four fractions using organic solvents with increasing polarity. A chronic 49-day dextran sulfate sodium (DSS)-induced colitis mice model, closely resembling human clinical conditions, was used to examine the effect of CDD-2103 on chronic colitis. To confirm the effect of CDD-2103 on macrophages in this chronic colitis model, adoptive macrophage transfer and CCL2 supplementation were conducted. The mechanisms of action of CDD-2103 were further elucidated utilizing bone marrow-derived macrophages (BMDMs). Transcriptome analysis was conducted to gain insights into the underlying mechanism of action of CDD-2103 in BMDMs. ResultsOur in vitro and in vivo findings demonstrated that the ethanol-enriched fraction of CDD-2103 exhibited significant anti-inflammatory effects, leading to the suppression of colitis severity. This effect was associated with diminished accumulation of colonic macrophages in the lamina propria of CDD-2103-intervened colitis mice. Specifically, CDD-2103 inhibited CCR2/L2-mediated proinflammatory macrophage infiltration into the colon without affecting macrophage proliferation. Mechanistically, CDD-2103 inhibited Fyn expression-mediated p38 MAPK activation and subsequently suppressed CCR2 expression in BMDMs. ConclusionsCollectively, our study supports the potential use of CDD-2103 to limit macrophage infiltration, thereby reducing inflammation during UC treatment. CDD-2103 and the components in the ethanolic fraction are promising candidates for the development of novel drugs for UC management. Additionally, our study underscores Fyn-mediated CCR2 expression as a potential therapeutic target for the management of UC.

Active Soil Organic Carbon Pools Decrease with Increased Time since Land-Use Transition from Rice Paddy Cultivation to Areca Nut Plantations under the Long-Term Application of Inorganic Fertilizer

Many croplands in the tropics of China have been converted over the last decades into areca nut plantations due to their high economic returns. This land-use transition was accompanied by changes in agricultural practices such as soil moisture regimes and fertilizer inputs, which may affect soil organic carbon (SOC) and its fractions, especially in tropical soils with low fertility and high nitrogen loss. Yet, how the time since land-use transition from rice paddy cultivation to areca nut plantations affects soil carbon dynamics and their underlying mechanisms in the tropics of China remains elusive. Here, areca nut plantation soils with different ages (2, 5, 10, 14, and 17 years) and paddy fields in the tropical region of China were investigated. The study result indicates that the contents of dissolved organic carbon (DOC), particulate organic carbon (POC), easily oxidized organic carbon (EOC), light organic carbon (LFOC), and microbial biomass carbon (MBC) decreased significantly with increased time since land-use transition from rice paddy cultivation to areca nut plantations. Similarly, the ratios of DOC/SOC, MBC/SOC, POC/SOC, LFOC/SOC, and EOC/SOC decreased significantly with increased time since land-use transition. Compared with the paddy soil, the carbon pool management index decreased by 36.6–76.7% under the areca nut plantations, concluding that increasing the time since land-use transition from rice paddy cultivation to areca nut plantations with high application rates of chemical fertilizers resulted in reduced soil active carbon fractions and SOC supply capacity. Therefore, agricultural practices such as the use of organic fertilizers should be applied to improve the soil’s ability to supply organic carbon in managed plantation ecosystems in the tropics of China.

Antidiabetic and antihyperlipidemic effects of crude fractions from Chlorophytum borivilianum root methanolic extract on streptozotocin induced diabetic rats and phytochemical investigation by LCMS analysis.

To evaluate the in vitro and in vivo pharmacological efficacy of the plant Chlorophytum borivilianum in diabetes and hyperlipidemia and to confine and describe the synthetic constituents from the roots that are in charge of the action. The present study was carried out to investigate the ethno-medical use of Chlorophytum borivilianum root methanolic extract as a potential anti-diabetic and antihyperlipidemic agent in STZ-induced diabetic rats. Extract was tested for in vitro and in vivo biological activities. Soxhlet extraction was carried out using methanol as a solvent, and TLC and column chromatography were used for fractionation. Liquid Chromatography and Mass Spectroscopic study confirmed the structures of isolated compounds. Chlorophytum borivilianum root methanolic extract showed the presence of phytoconstituents as Dihydrocapsaicin, Reserpine, Deserpidine, Biliverdin-IX-α, and Cassiamin C having a therapeutic effect. Dihydrocapsaicin was identified at RT 7.572 and the Chlorophytum borivilianum root chloroform methanolic extract fraction noticeably depleted increased blood glucose levels and had positive effects on altered lipid profile after administering a dose of 150 mg/kg orally compared with oral hypoglycemic drug metformin. All the results are dose-dependent. Active chloroform-methanol fraction from methanol extract showed the presence of anti-diabetic compound, Dihydrocapsaicin. The chloroform-methanol fraction from the methanolic extract of Chlorophytum borivilianum root can inhibit the parameters linked to diabetes and hyperlipidemia.

Новий метаболіт Myrothecium cinctum 910 трихотеценової природи та його біологічні властивості

Fungi of the Myrothecium genus are well-known producers of macrocyclic trichothecenes, characterized by the acute toxic effect on warm-blooded organisms and fungicidal action against a wide range of fungi. At the same time, as established by us earlier, M. cinctum (current name Striaticonidium cinctum) 910 shows a wide spectrum of antibacterial activity: along with antifungal activity against mycelial, yeast-like and phytopathogenic test-cultures it strongly inhibited the growth of gram-positive test-strains and to lesser extend – the growth of gram-negative and phytopathogenic bacteria. This strain also revealed significant phytotoxic potential suppressing the growth of green algae Chlorella strains. The aim of the work was to isolate, to purify and to obtain in crystalline form the biological active metabolites of M. cinctum 910 and to study their physicochemical characteristics and biological activity. Methods. To obtain biologically active metabolites in the purified crystalline form, the extraction followed by separation by column chromatography and recrystallization was used; physicochemical methods and microbiological tests were used for characterization of their properties. Results. Three substances which showed the wide spectrum of biological activity to indicator test-cultures were obtained in crystalline form: preparation МС910 with high antibiotic activity and preparations from fractions 8/2 and 9 with high antifungal and phytotoxic activities. Taking into account the data of spectroscopy in the UV-range of spectrum, of spectrums of antibiotic activity and of qualitative reaction with 4-(4-nitrobenzyl)pyridine, it can be concluded that active fractions  8/2 and  9 contain macrocyclic trichothecenes whereas the metabolite МС910 (which is proved by data of IR-spectroscopy) belongs to simple trichothecenes – substances like to macrocyclic trichothecenes but without macrocyclic fragment in their structure. Conclusions. Antibiotic properties of M. cinctum 910 are due to the complex of biological active metabolites with different biological and physicochemical properties. In view on the biological activity of MC910, it can be argued that it has atypical for trichothecenes, including macrocyclc trichothecenes, antibacterial properties as well as it is non-toxic and does not reveal dermatocidal activity in studied concentrations. Therefore, we assume that MC910 is new, not described earlier metabolite of trichothecene nature, which needs further studies. At the same time, active metabolites from fractions 8/2 and 9 are represented by macrocyclic trichothecenes, which is also indicated by the spectra of their biological activity.

Exploring the anti‐inflammatory ingredients and potential of golden buckwheat (Fagopyrum dibotrys) on the TLR4/NLRP3 pathway in acute lung injury

AbstractGolden buckwheat, also called Fagopyrum dibotrys (D. Don) H. Hara, is a plant of the genus Buckwheat in the buckwheat family. The aim of this study was to screen the bioactive ingredients of golden buckwheat extract and investigate the protective effect on acute lung injury (ALI). The ethyl acetate extract (EAE) was identified as the active fraction in LPS‐induced RAW264.7 cells, with gallic acid, proanthocyanidin B2, and epicatechin at 0.0563%, 0.3707%, and 0.3868%, respectively. At the same time, 20 compounds (mainly flavonoids and organic acids) were identified by UPLC‐Q‐Exactive Orbitrap‐HRMS in EAE. Furthermore, the EAE reduced lung histopathology scores in mice with ALI, decreased the dry‐to‐wet weight ratio of lung tissue, and significantly inhibited the concentrations of IL‐1β, TNFα, and IL‐6 in bronchoalveolar lavage fluid (BALF). It also reduced the number of leukocytes, decreased the activity of MPO in lung tissue, and inhibited the levels of TLR4/NLRP3 pathway mRNA and protein in lung tissue. Our study indicated that golden buckwheat as a source of functional food prevents or treats associated lung diseases by modulating the activation of the TLR4/NLRP3 signaling pathway.

Evolutionarily conserved ovarian fluid proteins are responsible for extending egg viability in salmonid fish

In contrast to most fishes, salmonids exhibit the unique ability to hold their eggs for several days after ovulation without significant loss of viability. During this period, eggs are held in the body cavity in a biological fluid, the coelomic fluid (CF) that is responsible for preserving egg viability. To identify CF proteins responsible for preserving egg viability, a proteomic comparison was performed using 3 salmonid species and 3 non-salmonid species to identify salmonid-specific highly abundant proteins. In parallel, rainbow trout CF fractions were purified and used in a biological test to estimate their egg viability preservation potential. The most biologically active CF fractions were then subjected to mass spectrometry analysis. We identified 50 proteins overabundant in salmonids and present in analytical fractions with high egg viability preservation potential. The identity of these proteins illuminates the biological processes participating in egg viability preservation. Among identified proteins of interest, the ovarian-specific expression and abundance in CF at ovulation of N-acetylneuraminic acid synthase a (Nansa) suggest a previously unsuspected role. We show that salmonid CF is a complex biological fluid containing a diversity of proteins related to immunity, calcium binding, lipid metabolism, proteolysis, extracellular matrix and sialic acid metabolic pathway that are collectively responsible for preserving egg viability.

Novel antifungal compound from Prunella vulgaris: An in-silico approach towards the discovery of potent inhibitor to combat phytopathogenic fungi

Phytopathogenic fungi wreak havoc on crops, causing substantial losses globally. Numerous chemical fungicides are employed to manage the pathogen, yet effectiveness is limited and resistance concerns are rising. The current study aimed to identify the antifungal compound from the spikes of a significant medicinal herb Prunella vulgaris subsp. hispida (Benth.) Hultén as an alternative to combat the fungi. In vitro, antifungal assay revealed that the ethyl acetate extract of P. vulgaris emerged as a potent inhibitor against selected fungal phytopathogens Fusarium oxysporum, Alternaria solani and Penicillium digitatum. Through chromatographic analysis, 32 compounds were identified from the active fraction of ethyl acetate extract. Notably, five compounds were reported for the first time from P. vulgaris. They include dichloroacetic acid, tridec-2-ynyl ester (1), phenol, 3,5-bis(1,1-dimethylethyl)- (2), 2-thiopheneacetic acid, 3-tetradecyl ester (3), 1H-1,2,4-triazol-3-amine, 1-ethyl- (4) and sulfurous acid, octadecyl 2-propyl ester (5). Compound 1 was identified as the most abundant. NMR studies confirmed the presence of compound 1, prompting its selection for in silico antifungal analysis. Various computational models showcased that Compound 1 achieved a high bioactive score and met the criteria for medicinal chemistry friendliness. Additionally, a molecular docking study revealed that compound 1 displayed a good binding affinity with the multiple target fungal enzymes such as scytalone dehydratase and cytochrome P450 which is a prime target for antifungal drugs. Remarkably, this study marks the first detection of compound 1’s effectiveness against phytopathogenic fungi. Therefore, compound 1 holds the potential to pave the way for the development of a potent novel fungicide.

Screening for Potential Antioxidants from &lt;i&gt;Chrozophora senegalensis&lt;/i&gt; Crude Extract

Numerous reports have indicated oxidative stress as the under-laying factor or associated with the onset or progression of numerous  human ailments and disorders such as dementia, Alzheimer disease, diabetes, macular degeneration and aging. Chrozophora  senegalensis (C. senegalensis) is a plant used in traditional medicine to cure both infectious and non-infectious diseases. This research  was aimed to carry out the antioxidant activities guided isolation and characterization of phytocompounds from C. senegalensis  methanol crude extract. The plant sample was extracted using maceration and concentrated on a rotary evaporator at 45oC. Three  different but complementary methods; Ferric reducing antioxidant power (FRAP), Nitric oxide and hydrogen peroxide assays were used  to evaluate the antioxidant activities of C. senegalensis crude extracts. Column chromatography was used to isolate and purify the  compounds from the most active n-hexane fraction and coded as CSC-1, CSC-2 and CSC-3. The compounds were characterized using NMR  (1H &amp; 13C), GC-MS, melting point determination and in comparison with literature. The percentage free radical scavenging activity  for FRAP, nitric oxide and hydrogen peroxide assays were 94.51%, 92.38% and 91.67% respectively while that of ascorbic acid standard  was 95.43±0.76%. The isolated compounds were identified as 1- hexacosanol (CSC-1), 1-triacontanol (CSC-2) and oleic acid (CSC-3). The  isolation and characterization of these bioactive compounds from C. senegalensis crude extract validates its usefulness in traditional  medicine practice.   

Major endothelial damage markers identified from hemadsorption filters derived from treated patients with septic shock - endoplasmic reticulum stress and bikunin may play a role.

In septic patients the damage of the endothelial barrier is decisive leading to circulatory septic shock with disseminated vascular coagulation, edema and multiorgan failure. Hemadsorption therapy leads to rapid resolution of clinical symptoms. We propose that the isolation of proteins adsorbed to hemadsorption devices contributes to the identification of mediators responsible for endothelial barrier dysfunction. Plasma materials enriched to hemadsorption filters (CytoSorb®) after therapy of patients in septic shock were fractionated and functionally characterized for their effect on cell integrity, viability, proliferation and ROS formation by human endothelial cells. Fractions were further studied for their contents of oxidized nucleic acids as well as peptides and proteins by mass spectrometry. Individual fractions exhibited a strong effect on endothelial cell viability, the endothelial layer morphology, and ROS formation. Fractions with high amounts of DNA and oxidized DNA correlated with ROS formation in the target endothelium. In addition, defined proteins such as defensins (HNP-1), SAA1, CXCL7, and the peptide bikunin were linked to the strongest additive effects in endothelial damage. Our results indicate that hemadsorption is efficient to transiently remove strong endothelial damage mediators from the blood of patients with septic shock, which explains a rapid clinical improvement of inflammation and endothelial function. The current work indicates that a combination of stressors leads to the most detrimental effects. Oxidized ssDNA, likely derived from mitochondria, SAA1, the chemokine CXCL7 and the human neutrophil peptide alpha-defensin 1 (HNP-1) were unique for their significant negative effect on endothelial cell viability. However, the strongest damage effect occurred, when, bikunin - cleaved off from alpha-1-microglobulin was present in high relative amounts (>65%) of protein contents in the most active fraction. Thus, a relevant combination of stressors appears to be removed by hemadsorption therapy which results in fulminant and rapid, though only transient, clinical restitution.

Purification and structural characterization of APTs, a novel antifungal peptide produced by Lactobacillus paracasei ALAC-4, and its inhibitory effect on Candida albicans

Fungal contamination poses significant food safety risks. Lactobacillus paracasei ALAC-4, originally isolated from traditional fermented foods in Inner Mongolia, exhibits antifungal properties. Here, we isolated and purified the active fraction from ALAC-4 to derive the antifungal peptide, demonstrating its antifungal efficacy against Candida albicans. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) revealed severe deformation of C. albicans cell surfaces upon exposure to the antifungal peptide, resulting in membrane rupture and pore formation. Simultaneously, a decrease in Zeta potential and K+ ion leakage occurred on the C. albicans cell surface. Treatment with antifungal peptides reduced the hydrophobicity of the cell surface, indicating decreased adsorption rates of C. albicans cells. Additionally, LC-MS/MS analysis identified the antifungal peptide APTs with a molecular mass of 423.18 Da and an amino acid sequence of S-G-G-G-F, representing a novel and previously unreported antifungal peptide. Circular dichroism analysis indicated that the secondary structure of APTs primarily comprised α-helices. This study underscores the potential of APTs as a natural biopreservative for food preservation.

Saturated Iso-Type Fatty Acids from the Marine Bacterium Mesoflavibacter zeaxanthinifaciens with Anti-Trypanosomal Potential.

Chagas disease is a Neglected Tropical Disease with limited and ineffective therapy. In a search for new anti-trypanosomal compounds, we investigated the potential of the metabolites from the bacteria living in the corals and sediments of the southeastern Brazilian coast. Three corals, Tubastraea coccinea, Mussismilia hispida, Madracis decactis, and sediments yielded 11 bacterial strains that were fully identified by MALDI-ToF/MS or gene sequencing, resulting in six genera-Vibrio, Shewanella, Mesoflavibacter, Halomonas, Bacillus, and Alteromonas. To conduct this study, EtOAc extracts were prepared and tested against Trypanosoma cruzi. The crude extracts showed IC50 values ranging from 15 to 51 μg/mL against the trypomastigotes. The bacterium Mesoflavibacter zeaxanthinifaciens was selected for fractionation, resulting in an active fraction (FII) with IC50 values of 17.7 μg/mL and 23.8 μg/mL against the trypomastigotes and amastigotes, respectively, with neither mammalian cytotoxicity nor hemolytic activity. Using an NMR and ESI-HRMS analysis, the FII revealed the presence of unsaturated iso-type fatty acids. Its lethal action was investigated, leading to a protein spectral profile of the parasite altered after treatment. The FII also induced a rapid permeabilization of the plasma membrane of the parasite, leading to cell death. These findings demonstrate that these unsaturated iso-type fatty acids are possible new hits against T. cruzi.