Evaluation of antifungal potentials of Albizia kalkora extract as a natural fungicide: In vitro and computational studies.

The antifungal bioactivity potential of the organic extract of silk tree (Albizia kalkora) was investigated in the current study. The crude extracts of A. kalkora and methanol, n-hexane, chloroform, and ethyl acetate fractions were prepared. The antifungal activity of obtained fractions of A. kalkora was studied at different concentrations ranging from 0.39-50 µg/mL. Dimethyl sulfoxide (DMSO) was taken as a toxicity control, whereas thiophanate methyl (TM) as a positive control. All the fractions significantly reduced the FOL growth (methanolic: 9.49-94.93 %, n-hexane: 11.12-100 %, chloroform: 20.96-91.41 %, and ethyl acetate: 18.75-96.70 %). The n-hexane fraction showed 6.25 µg/mL MIC as compared to TM with 64 µg/mL MIC. The non-polar (n-hexane) fraction showed maximum antifungal bioactivity against FOL in comparison with chloroform, methanol, and ethyl acetate fractions. GC/MS analysis exhibited that the n-hexane fraction contained hexadecanoic acid, 9,12,15-octadecatrienoic acid, 9,12-octadecadienoic acid, bis(2-ethylhexyl) phthalate, methyl stearate, and [1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylic acid. The results of in vitro antifungal inhibition were further reinforced by molecular docking analysis. Five virulence proteins of FOL i.e., pH-responsive PacC transcription factor (PACC), MeaB, TOR; target of rapamycin (FMK1), Signal transducing MAP kinase kinase (STE-STE7), and High Osmolarity Glycerol 1(HOG1) were docked with identified phytocompounds in the n-hexane fraction by GC/MS analysis. MEAB showed maximum binding affinities with zinnimide (-12.03 kcal/mol), HOG1 and FMK1with α-Tocospiro-B (-11.51 kcal/mol) and (-10.55 kcal/mol) respectively, STE-STE7 with docosanoic acid (-11.31 kcal/mol), and PACC with heptadecanoic acid (-9.88 kcal/mol) respectively with strong hydrophobic or hydrophilic interactions with active pocket residues. In conclusion, the n-hexane fraction of the A. kalkora can be used to manage FOL.

Effect of Mosquitocidal, histopathological alteration and non target effects of Sigesbeckia orientalis L. on Anopheles stephensi Liston, Culex quinquefasciatus say and Aedes aegypti L.

Diseases transmitted by vectors have a significant collision on society and community health, particularly in tropical and subtropical regions, where they can cause large-scale outbreaks shortly after initial transmission. The intent of this investigation was to study the plant extract derived from Sigesbeckia orientalis L. in controlling the immature stages of Anopheles, Culex and Aedes mosquitoes, while also considering its potential toxicity to ecosystems. The immature stages were exposed to different extracts (62.5-500 ppm), and the mortality of larvae and pupae, as well as ovicidal activity, were noted after 24 and 120 h of the experiment. The hexane and ethyl aceate extract of S. orientalis presented 100% ovicidal activity against the eggs of Anopheles, Aedes and Culex at 500 ppm concentration after 5 days of treatment. The hexane and ethylacetate extracts presented strong larvicidal activity with LC50 values of 215.7, 332.0, 197.4 and 212.6, 694.9 and 201.7 ppm against treated mosquitoes at 24 h, respectively. The same extract also presented promising pupicidal activity. The LC50 values of hexane extract were 219.6, 353.6, 194.2 and LC50 values of ethyl acetate were 257.6, 387.8 and 259.07 ppm against early stage pupae of three vector mosquitoes, respectively. The extracts from S. orientalis had strong inhibitory activity against growth and development of mosquitoes. SI/PSF values showed that the extracts of S. orientalis did not harm Poecilia reticulata, Diplonychus indicus (Water bug), Gambusia affinis and dragon fly nymph at tested concentrations. Furthermore, examinations of histopathology and growth disruption revealed significant damage to the midgut cells in the treated larvae. The formulations utilizing hexane and ethyl acetate extracts exhibited potent activity without posing any toxicity towards non-target organisms. This study clearly indicated that hexane and ethylacetate extracts showed promising results against treated mosquitoes. The present study documents the first report of the extracts from S. orientalis and they can be further assessed to identify compounds for application purposes.

Hexane Fraction of Adenophora triphylla var. japonica Root Extract Inhibits Angiogenesis and Endothelial Cell-Induced Erlotinib Resistance in Lung Cancer Cells.

The aim of this study was to investigate the anti-angiogenic effects of the hexane fraction of Adenophora triphylla var. japonica root extract (HAT) and its influence on the development of erlotinib resistance in human lung cancer cells. HAT significantly reduced the migration, invasion, and tube formation of human umbilical vein endothelial cells (HUVECs). The phosphorylation levels of vascular endothelial growth factor receptor 2 (VEGFR2) and its downstream molecules were decreased via HAT, indicating its anti-angiogenic potential in endothelial cells (ECs). A docking analysis demonstrated that ß-sitosterol and lupeol, representative components of HAT, exhibit a high affinity for binding to VEGFR2. In addition, conditioned media from HAT-pretreated H1299 human lung cancer cells attenuated cancer-cell-induced chemotaxis of HUVECs, which was attributed to the decreased expression of angiogenic and chemotactic factors in H1299 cells. Interestingly, co-culture of erlotinib-sensitive PC9 human lung cancer cells with HUVECs induced erlotinib resistance in PC9 cells. However, co-culture with HAT-pretreated HUVECs partially restored the sensitivity of PC9 cells to erlotinib. HAT inhibited the development of erlotinib resistance by attenuating hepatocyte growth factor (HGF) production by ECs. Taken together, our results demonstrate that HAT exerts its anticancer effects by regulating the crosstalk between ECs and lung cancer cells.

Modulation of quorum sensing and biofilm of Gram-negative bacterial pathogens by Cinnamomum zeylanicum L.

The development of antibiotic resistant microbial pathogens has become a global health threat and a major concern in modern medicine. The problem of antimicrobial resistance (AMR) has majorly arisen due to sub-judicious use of antibiotics in health care and livestock industry. A slow progress has been made in last two decades in discovery of new antibiotics. A new strategy in combatting AMR is to modulate or disarm the microbes for their virulence and pathogenicity. Plants are considered as promising source for new drugs against AMR pathogens. In this study, fraction-based screening of the Cinnamomum zeylanicum extract was performed followed by detailed investigation of antiquorum sensing and antibiofilm activities of the most active fraction that is, C. zeylanicum hexane fraction (CZHF). More than 75% reduction in violacein pigment of C. violaceum 12472 was overserved. CZHF successfully modulated the virulence of Pseudomonas aeruginosa PAO1 by 60.46%-78.35%. A similar effect was recorded against Serratia marcescens MTCC 97. A broad-spectrum inhibition of biofilm development was found in presence of sub-MICs of CZHF. The colonization of bacteria onto the glass coverslips was remarkably reduced apart from the reduction in exopolymeric substances. Alkaloids and terpenoids were found in CZHF. GC/MS analysis revealed the presence of cinnamaldehyde dimethyl acetal, 2-propenal, coumarin, and α-copaene as major phytocompounds. This study provides enough evidence to support potency of C. zeylanicum extract in targeting the virulence of Gram -ve pathogenic bacteria. The plant extract or active compounds can be developed as successful drugs after careful in vivo examination to target microbial infections. RESEARCH HIGHLIGHTS: Hexane fraction of Cinnamomum zeylanicum is active against QS and biofilms. The broad-spectrum antibiofilm activity was further confirmed by microscopic analysis. Dimethyl acetal, 2-propenal, coumarin, α-copaene, and so forth are major phytocompounds.

Exploring the bioactive potential leaves of Sphaeranthus indicus: Targeting immature stages of Aedes aegypti and Culex quinquefasciatus mosquito vectors through bioassay-guided screening and fraction isolation.

Mosquito control is vital for combating mosquito-borne diseases, but concerns exist regarding the use of synthetic insecticides. This study aimed to explore eco-friendly alternatives derived from natural sources. The larvicidal, pupicidal, and ovicidal activities of various fractions obtained from the hexane leaf extract of Sphaeranthus indicus were investigated against two important mosquito vectors, Aedes aegypti and Culex quinquefasciatus. S. indicus leaves were extracted with hexane and column chromatography was performed with hexane, ethyl acetate, methanol, and their mixtures as eluents. Among the ten fractions (F1-F10) evaluated, fraction 'F-4' exhibited significant activity against third instar larvae, pupae, and eggs of both mosquito species, closely followed by 'F-5' . At a concentration of 10 ppm, 'F-4' achieved 100% mortality in larvae and displayed LC50 values of 5.08 ppm and 5.03 ppm for Ae. aegypti and Cx. quinquefasciatus larvae, respectively. The LC50 values for pupae were 6.12 ppm and 5.83 ppm for Ae. aegypti and Cx. quinquefasciatus, respectively. Regarding ovicidal activity, 'F-4' demonstrated percentages ranging from 63.2% to 64.8% against Ae. aegypti and Cx. quinquefasciatus eggs, respectively. These findings underscore the potent larvicidal, pupicidal, and ovicidal effects of fraction 'F-4' from S. indicus against the targeted mosquito species. Further research is warranted to identify the active compounds responsible for these effects and explore practical applications for sustainable mosquito control strategies.

The Hexane Extract of Citrus sphaerocarpa Ameliorates Visceral Adiposity by Regulating the PI3K/AKT/FoxO1 and AMPK/ACC Signaling Pathways in High-Fat-Diet-Induced Obese Mice.

Obesity is an emerging global health issue with an increasing risk of disease linked to lifestyle choices. Previously, we reported that the hexane extract of Citrus sphaerocarpa (CSHE) suppressed lipid accumulation in differentiated 3T3-L1 adipocytes. In this study, we conducted in vivo experiments to assess whether CSHE suppressed obesity in zebrafish and mouse models. We administered 10 and 20 µg/mL CSHE to obese zebrafish juveniles. CSHE significantly inhibited visceral fat accumulation compared to untreated obese fish. Moreover, the oral administration (100 µg/g body weight/day) of CSHE to high-fat-diet-induced obese mice significantly reduced their body weight, visceral fat volume, and hepatic lipid accumulation. The expression analyses of key regulatory genes involved in lipid metabolism revealed that CSHE upregulated the mRNA expression of lipolysis-related genes in the mouse liver (Pparα and Acox1) and downregulated lipogenesis-related gene (Fasn) expression in epididymal white adipose tissue (eWAT). Fluorescence immunostaining demonstrated the CSHE-mediated enhanced phosphorylation of AKT, AMPK, ACC, and FoxO1, which are crucial factors regulating adipogenesis. CSHE-treated differentiated 3T3L1 adipocytes also exhibited an increased phosphorylation of ACC. Therefore, we propose that CSHE suppresses adipogenesis and enhances lipolysis by regulating the PI3K/AKT/FoxO1 and AMPK/ACC signaling pathways. These findings suggested that CSHE is a promising novel preventive and therapeutic agent for managing obesity.

Mosquito larvicidal, pupicidal and ovicidal effects of the extracts of Peltophorum pterocarpum flowers on Aedes aegypti and Culex quinquefasciatus.

Mosquitoes are the vectors of several diseases like dengue, chikungunya, malaria etc. The three important mosquito species in India are Aedes aegypti, Culex quinquefasciatus and Anopheles stephensi. Several plant extracts have been tested for their anti-mosquito activity. In this dissertation, the larvicidal, pupicidal and ovicidal activities of the successive hexane, chloroform and methanol extracts of Peltophorum pterocarpum (Fabaceae) on mosquitoes are reported. The larvicidal activity of those extracts on both Aedes aegypti and Culex quinquefasciatus mosquitoes was found to be in a decreasing order of hexane, methanol and chloroform - the LC50 values for these extracts were 111.81 and 104.84 ppm, 121.92 and 121.56 ppm, 357.2 and 352.0 ppm respectively. Their pupicidal activity on both mosquitoes was found in the order of methanol, chloroform and hexane - the LC50 values for these extracts being 172.8 and 162.35 ppm, 425.8 respectively. Their ovicidal activity on both mosquitoes was found to be very low, even at the higher concentration of 500 ppm. This is the first report on the effect of the extracts of Peltophorum pterocarpum flowers on the two mosquito species of Aedes aegypti, Culex quinquefasciatus. In the present work, the results showed that the hexane extract had the highest larvicidal activity, while methanol showed the highest pupicidal activity besides larvicidal activity. Hence, the methanol extract holds promise to be used as mosquitocidal agent against the above two vectors.

Induction of Apoptosis and Modulation of Caspase Activity on MCF-7 Human Breast Cancer Cells by Bioactive Fractionated Cocoa Leaf Extract.

BACKGROUND: The objective of this study was to investigate the potential anti-proliferative activities of a methanolic extract of cocoa leaves (CL) obtained through sequential partition and fractionation against MCF-7 breast cancer cells.  Methods: The methanolic extract of CL was partitioned in three separated solvents (hexane, dichloromethane, and methanol). Hexane partition was the most potent against MCF-7 cells growth with the lowest IC50 value. Then, it was subjected to two fractionation procedures, resulting in the identification of the CL bioactive fraction (II-F7) with potent toxicity against MCF-7 cells. RESULTS: Further investigation into CL bioactive fraction (II-F7) revealed significant dose-dependent growth inhibitory effects on MCF-7 cells, which were attributed to the induction of apoptosis, as evidenced by the presence of apoptotic bodies, fragmented DNA, and disruption of mitochondrial membrane potential. Additionally, treatment with CL bioactive fraction (II-F7) upregulated the expression of pro-apoptotic genes (DDIT3, GADD45G and HRK) and significantly increased the activities of caspase-8 and caspase-9. CONCLUSION: Overall, this study suggests that bioactive fraction (II-F7) from CL extract has significant and selective cytotoxicity against MCF-7 cells through inducing apoptosis and has potential as a therapeutic agent for breast cancer treatment.

Decellularization: Leveraging a Tissue Engineering Technique for Food Production.

The edible nature of many plants makes leaves particularly useful as scaffolds for the development of cultured meat, where animal tissue is grown in the laboratory setting. Recently, we demonstrated that decellularized spinach leaves can serve as scaffolds to grow and differentiate cells for cultured meat products. However, conventional decellularization methods use solutions that are not considered safe for use in food, such as organic solvents (hexanes) and detergents (triton X-100 (TX100)). This study modified decellularization protocols to incorporate detergents that are regulated (REG) by the United States Food and Drug Administration (FDA) for use in food, such as Polysorbate 20 (PS20), and eliminates the use of hexanes for cuticle removal. Spinach leaves were decellularized with sodium dodecyl sulfate and then with either TX100 (control) or PS20. The average DNA content for TX100 samples and PS20 samples was similar (1.3 ± 0.07 vs 1.3 ± 0.05 ng/mg; TX100 vs PS20, p = ns). The importance of cuticle removal was tested by removing hexanes from the protocol. Groups that included the cuticle removal step exhibited an average reduction in DNA content of approximately 91.7%, and groups that omitted the cuticle removal step exhibited an average reduction of approximately 90.3% (p = ns), suggesting that the omission of the cuticle removal step did not impede decellularization. Lastly, primary bovine satellite cells (PBSCs) were cultured for 7 days (d) on the surface of spinach leaves decellularized using the REG protocol. After the 7 d incubation period, PBSCs grown on the surface of REG scaffolds had an average viability of approximately 97.4%. These observations suggest that the REG protocol described in this study is an effective decellularization method, more closely adhering to food safety guidelines, that could be implemented in lab grown meat and alternative protein products.

Anti-cancer Effect and Active Phytochemicals of Houttuynia cordata Thunb. against Human Breast Cancer Cells.

BACKGROUND: Houttuynia cordata Thunb (HCT) is a medicinal herb used in Southeast Asia. Aim of this work: This study aimed at investigating the cytotoxicity of this plant extract and fractions towards human breast cancer MDA-MB-231 and MCF-7 cells. HCT's phytoactive compounds are determined. MATERIALS AND METHODS: Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The mode of cell death was measured by staining with annexin V-FITC and propidium iodide (PI) employing flow cytometry technique. The oxidative stress was measured by using 2',7'-dihydrodichlorofluorescein diacetate (DCFH-DA) and dihydroethidium (DHE+) fluorescent probes and using a fluorescence microplate reader. HCT phytochemicals were characterized by high performance liquid chromatography (HPLC). RESULTS: The proliferation of MDA-MB-231 and MCF-7 cells was dramatically decreased by the crude extract and individual fraction of HCT. Ethyl acetate was the solvent fraction with the highest toxicity against MCF-7 cells, followed by dichloromethane, crude, and hexane fractions, respectively, whereas in MDA-MB231 cells, dichloromethane, crude, hexane, and ethyl acetate fractions each had the strongest impact, respectively. The methanol fraction had no effect on either cell line up to 200 µg/ml. The extract and fractions were less harmful to the NIH3T3 normal murine fibroblast cell line. The mode of both cell death was apoptosis evidenced by the increase of cell population stained with annexin V-FITC and PI. The fluorescence probes of both DCFH-DA and DHE in MDA-MB-231 cell line were enhanced. Phenolic acids included chlorogenic acid (CA), gallic acid (GA), transcoumaric acid (TCA), vanillic acid (VA), and syringic acid (SA), as well as flavonoids like quercetin and rutin, were identified as the active phytochemicals in the crude and fractions by using HPLC method. CONCLUSION: MDA-MB-231cells underwent apoptosis via oxidative stress when induced with HCT hexane fraction. Phenolic acids and flavonoids were identified in HCT's extract and fractions.

Asparagus falcatus L. (Asparagaceae) leaf extracts attenuate doxorubicin-induced renal toxicity via antioxidant, anti-inflammatory, and anti-apoptotic pathways.

The search for therapeutic agents that improve kidney function against doxorubicin-induced renal toxicity is important. Herein, the potential nephroprotective activity by Asparagus falcatus L. (AF, Asparagaceae) leaf extracts against doxorubicin-induced renal toxicity (5 mg/kg, ip) in Wistar rats (n = 6/group) after oral administration of hexane (55 mg/kg), ethyl acetate (35 mg/kg), butanol (75 mg/kg), and aqueous (200 mg/kg) extracts of AF for 28 consecutive days was investigated. It was noticed that the treatment with the selected extracts of AF significantly attenuated doxorubicin-induced elevations of serum creatinine, urea nitrogen, ß2-microglobulin, cystatin C, and proteinuria in experimental rats. The histology showed attenuation of the features of acute tubular injury. Treatment regimens significantly reversed the doxorubicin-induced reduction in total antioxidant status, glutathione peroxidase, and glutathione reductase activity in renal tissue homogenates. A suppression in lipid peroxidation was noted with hexane, ethyl acetate, and butanol extracts of AF. Moreover, a reduction in the concentration of the pro-inflammatory mediator TNF-α (p < 0.05), and immunohistochemical expression of COX-2 were observed. The immunohistochemical expression of pro-apoptotic Bax protein was decreased and the anti-apoptotic BCL-2 was increased in renal tissues following the treatments. In conclusion, it was revealed that, hexane, ethyl acetate, butanol, and aqueous extracts of AF attenuate doxorubicin-induced renal toxicity in Wistar rats through antioxidant, anti-inflammatory, and anti-apoptotic pathways. The plant, AF could be recommended as a promising therapeutic agent to minimize renal toxicity induced by doxorubicin in cancer patients, however, subsequent clinical trials are warranted.

Toxicity of Ocimum americanum L. extracts against Aedes aegypti larvae (Diptera: Culicidae) and their impact on insecticide resistance.

Mosquitoes are a key threat to millions of people worldwide. They spread the pathogens that cause deadly diseases among humans and animals. Synthetic pesticides are the best agents to control mosquitoes, but they cause several problems for the environment as well as public health. Continuous usage of commonly available insecticides develops multiple resistances among pests. In search of alternatives to synthetic pesticides, botanicals could be one of the best alternatives to control mosquitoes. The present study explores the insecticidal activity of Ocimum americanum against Aedes aegypti larvae and their effect on detoxification enzymes. Leaves of O. americanum were sequentially extracted using hexane, chloroform, and methanol. Among these, hexane extract showed 100% larvicidal activity at 1 g/L concentration for 24 h and the LC50 value was 0.3 g/L. The phytochemical screening of hexane extract was performed through gas chromatography-mass spectrometry analysis, which showed 27 compounds. The major compounds are squalene (13.03%), camphor (9.77%), and 1-Iodohexadecane (8.02%). The toxicity of active hexane extract was tested against third instar larvae of Chironomus costatus (nontarget organism). Results revealed less toxicity (12.2%) at 1 g/L concentration on the nontarget organism. The enzyme activity of acetylcholinesterase and ß-carboxylesterase was significantly inhibited by the hexane extract. The present study reveals the insecticidal potential of O. americanum with minimum effects on nontarget organisms. The O. americanum extract inhibited the activity of A. aegypti's major insecticide-resistant enzymes. O. americanum could be one of the best alternatives to controlling mosquitoes.

Hepatoprotective Role Of N-Hexane Fraction And Crude Methanolic Extract Of Withania Coagulans In Balb C Mice.

Background: Withania coagulans is one of the most important medicinal herbs due to its wide range of biological activities. The aim of this study was to compare the hepatoprotective activity of crude methanolic extract versus n- hexane fraction of fruit of Withania coagulans in CCl4 induced liver toxicity. Methods: This study was done on 36 Balb c mice in Department of Anatomy of Khyber Medical College Peshawar. Results: The mean serum of Group 1(control Group) was 33.41±1.82U/L, for Group 2 (CCl4 treated Group) was 89.01±7.51 U/L, for Group 3(low dose Group) was 49.91±3.48 U/L and for Group 4 (High dose Group) was 50.86±4.87 U/L. There was significant difference in the readings of Group1 and Group 2 which indicated CCL4 induced hepatotoxicity in two groups. There was significant difference in the values of Group 3 and Group 4 (p value .000), showing the hepatotoxicity in these Groups was further enhanced. The mean AST at the end of six weeks for Group1 was 26.80±3.21U/L, for Group 2 was 149.01±13.63U/L, for Group 3 (including both low doses) was 70.81±7.92U/L and for Group 4 (High doses group) was 51.01±11.05U/. Conclusion: Withania coagulans both fractions have hepatoprotective effect against CCL4 induced hepatic toxicity in high and low doses in Balb c mice.

First Evidence of Gastroprotection by Schinus molle: Roles of Nitric Oxide, Prostaglandins, and Sulfhydryls Groups in Its Mechanism of Action.

Schinus molle is a plant traditionally used in Mexico to treat gastric disorders. However, no scientific evidence has been reported on its gastroprotective effect. The aim of the current contribution was to conduct a bioassay-guided study on S. molle to evaluate its gastroprotective activity in a model of Wistar rats given ethanol orally to induce gastric lesions. The hexane and dichloromethane extracts from the tested plant showed over 99% gastroprotection at a dose of 100 mg/kg. From the hexane extract, two of the three fractions (F1 and F2) afforded over 99% gastroprotection. The F1 fraction was subjected to column chromatography, which revealed a white solid. Based on the ESI-MS analysis, the two main compounds in this solid were identified. The predominant compound was probably a triterpene. This mixture of compounds furnished about 67% gastroprotection at a dose of 100 mg/kg. Pretreatment with L-NAME, indomethacin, and NEM was carried out to explore the possible involvement of nitric oxide, prostaglandins, and/or sulfhydryl groups, respectively, in the gastroprotective activity of the white solid. We found evidence for the participation of all three factors. No antisecretory activity was detected (tested by pylorus ligation). In conclusion, evidence is herein provided for the first time of the gastroprotective effect of S. molle.

Insecticidal effects of some selected plant extracts against Anopheles stephensi (Culicidae: Diptera).

BACKGROUND: The use of synthetic insecticides against mosquitoes may lead to resistance development and potential health hazards in humans and the environment. Consequently, a paradigm needs to shift towards the alternative use of botanical insecticides that could strengthen an insecticide resistance management programme. This study aimed to assess the insecticidal effects aqueous, hexane, and methanol crude leaf extracts of Calpurnia aurea, Momordica foetida, and Zehneria scabra on an insectary colony of Anopheles stephensi larvae and adults. METHODS: Fresh leaves of C. aurea, M. foetida and Z. scabra were collected and dried, then separately ground to powder. Powdered leaves of test plants were extracted using sonication with aqueous, hexane, and methanol solvents. The extracts were concentrated, and a stock solution was prepared. For comparison, Temephos (Abate®) and control solutions (a mixture of water and emulsifier) were used as the positive and negative controls, respectively. Different test concentrations for the larvae and the adults were prepared and tested according to WHO (2005) and CDC (2010) guidelines to determine lethal concentration (LC) values. Mortality was observed after 24 h exposure. The statistical analyses were performed using Statistical Package for the Social Sciences (SPSS) software (Kruskal-Wallis test) and R software (a generalized linear model was used to determine LC50 and LC90 values of the extracts). RESULTS: The lowest LC50 values were observed in aqueous extracts of M. foetida followed by Z. scabra extract and C. aurea leaves at 34.61, 35.85, and 38.69 ppm, respectively, against the larvae. Larval mortality was not observed from the hexane extracts and negative control, while the standard larvicide (temephos) achieved 100% mortality. Further, the adulticidal efficacy was greatest for aqueous extract of Z. scabra with LC50 = 176.20 ppm followed by aqueous extract of C. aurea (LC50 = 297.75 ppm). CONCLUSION: The results suggest that the leaf extracts of the three test plants have the potential of being used for the control of vector An. stephensi larvae and adult instead of synthetic mosquitocides. Further studies need to be conducted to identify the active ingredients and their mode of action.

Assessment of Antibacterial Efficacy of Callistemon viminalis (Sol. ex Gaertn.) G. Don against Some Isolates Obtained from Urinary Tract Infections.

This study aimed to examine the antibacterial effects of constituents obtained from Callistemon viminalis leaves. This goal was achieved by using three organic solvents, namely Ethanol, Ethyl acetate, and Hexane to prevent the growth of the causative urinary tract infections isolates, such as Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, and Proteus sp. in Iraq. The C. viminalis fresh leaves collected from different regions of Hillah City, during March 2020, were classified according to the taxonomic features of Iraqi Flora. Extractions were completed by a method of digestion and then the stock solution of 200 mg/mL was prepared in 10% of Dimethylsulfoxide. A Millipore filter (0.22 µm) was used for the sterilization of all the extracts used in this study. Agar well diffusion method was utilized to test the antibacterial effects of the constituents separated from the dried leaves of C. viminalis against the urinary tract infection bacteria at three concentrations of 50, 100, and 200 mg/mL for each extracted constitute by the three different solvents. Dimethylsulfoxide 10% and the meropenem were utilized as the negative and positive controls. Constituents separated by ethanol solvent at 200 mg/mL exhibited significant supremacy (P≤0.05) over the meropenem against Proteus sp. isolate, and exhibited the same significant difference (P≤0.05), compared to the meropenem drug against E. coli. Constituents extracted by Ethyl acetate organic solvent at a concentration of 200 mg/mL exhibited a similarly significant effect (P≤0.05), compared to the meropenem against Proteus sp. isolate. However, the hexane extract was the least effective among the other solvents utilized in this study. The results of the current study revealed that constituents in the leaves of C. viminalis could be considered a valuable herbal remedy for controlling urinary tract infections pathogenic bacteria.

Permixon®, hexane-extracted Serenoa repens, inhibits human prostate and bladder smooth muscle contraction and exerts growth-related functions in human prostate stromal cells.

AIMS: Recently, the European Association of Urology recommended hexane-extracted fruit of Serenoa repens (HESr) in their guidelines on management of non-neurogenic male lower urinary tracts symptoms (LUTS). Despite previously lacking recommendations, Permixon® is the most investigated HESr in clinical trials, where it proved effective for male LUTS. In contrast, underlying mechanisms were rarely addressed and are only marginally understood. We therefore investigated effects of Permixon® on human prostate and detrusor smooth muscle contraction and on growth-related functions in prostate stromal cells. MAIN METHODS: Permixon® capsules were dissolved using n-hexane. Contractions of human prostate and detrusor tissues were induced in organ bath. Proliferation (EdU assay), growth (colony formation), apoptosis and cell death (flow cytometry), viability (CCK-8) and actin organization (phalloidin staining) were studied in cultured human prostate stromal cells (WPMY-1). KEY FINDINGS: Permixon® inhibited α1-adrenergic and thromboxane-induced contractions in prostate tissues, and methacholine-and thromboxane-induced contractions in detrusor tissues. Endothelin-1-induced contractions were not inhibited. Neurogenic contractions were inhibited in both tissues in a concentration-dependent manner. In WPMY-1 cells, Permixon® caused concentration-dependent breakdown of actin polymerization, inhibited colony formation, reduced cell viability, and proliferation, without showing cytotoxic or pro-apoptotic effects. SIGNIFICANCE: Our results provide a novel basis that allows, for the first time, to fully explain the ubiquitous beneficial effects of HESr in clinical trials. HESr may inhibit at least neurogenic, α1-adrenergic and thromboxane-induced smooth muscle contraction in the prostate and detrusor, and in parallel, prostate stromal cell growth. Together, this may explain symptom improvements by Permixon® in previous clinical trials.

Biological Evaluation of 3-Azaspiro[Bicyclo[3.1.0]Hexane-2,5'-Pyrimidines] as Potential Antitumor Agents.

A series of heterocyclic compounds containing spirofused barbiturate and 3-azabicyclo[3.1.0]hexane frameworks have been studied as potential antitumor agents. Antiproliferative activity of products was screened in human erythroleukemia (K562), T lymphocyte (Jurkat), and cervical carcinoma (HeLa) as well as mouse colon carcinoma (CT26) and African green monkey kidney epithelial (Vero) cell lines. The most effective among the screened compounds show IC50 in the range from 4.2 to 24.1 µM for all tested cell lines. The screened compounds have demonstrated a significant effect of the distribution of HeLa and CT26 cells across the cell cycle stage, with accumulation of cells in SubG1 phase and induced apoptosis. It was found, using a confocal microscopy, that actin filaments disappeared and granular actin was distributed diffusely in the cytoplasm of up to 90% of HeLa cells and up to 64% of CT26 cells after treatment with tested 3-azaspiro[bicyclo [3.1.0]hexane-2,5'-pyrimidines]. We discovered that the number of HeLa cells with filopodium-like membrane protrusions was reduced significantly (from 91% in control cells to 35%) after treatment with the most active compounds. A decrease in cell motility was also noticed. Preliminary in vivo experiments on the impact of the studied compounds on the dynamics of CT26 tumor growth in Balb/C mice were also performed.

Rapeseed meal processing and dietary enzymes modulate excreta inositol phosphate profile, nutrient availability, and production performance of broiler chickens.

This study aimed to assess the effect of rapeseed meal (RSM) processing method, where solvent extraction occurred under standard industry conditions (ST) or cold-pressed hexane extraction was employed (MT), and exogenous enzyme supplementation (phytase [PHY] and xylanase [XYL]) alone or in combination on key nutritional factors of broiler chickens. A randomized control experiment was performed using 144 male Ross 308 broilers in a 2 × 2 × 3 factorial arrangement. Three diets including a nutritionally complete wheat-based basal diet (BD), a diet containing 200 g/kg of RSM extracted under ST and another diet containing 200 g/kg of RSM extracted under MT were produced. Each diet was then split into 4 parts and was fed as is, or supplemented with PHY at 1,500 FTU/kg or XYL at 16,000 BXU/kg, alone or in combination, resulting in 12 diets in total. Response criteria: feed intake (FI), weight gain (WG), and feed conversion ratio (FCR), from 7 to 21 d age, AMEn, retention coefficients for dry matter (DMR), nitrogen (NR), fat (FR), and the profile of inositol phosphate esters (IP2-6) and myo-inositol (MI) in excreta. Diets containing MT had higher AMEn compared to ST diets (P < 0.05). There was RSM by PHY interaction for FI, as only birds fed MT diet responded to PHY supplementation with reduced FI and FCR (P < 0.001). Feeding XYL reduced overall FI and FCR (P < 0.05). Feeding PHY reduced IP6 and increased MI in excreta (P < 0.001). Feeding XYL and PHY in combination reduced MI in excreta compared to PHY only (P = 0.05). Compared to BD, birds fed RSM diets had an increased IP6 (P < 0.05) and MI concentration in excreta (P < 0.01). This may be due to IP ester differences in RSM and BD.

Fractionation of hexane extracts from Achyrocline satureioides and their biological activities against Paenibacillus larvae.

Previous studies carried out in our laboratory described the antimicrobial activity of the whole hexanic extract (HE) of Achyrocline satureioides (Lam.) DC against Paenibacillus larvae, the causal agent of American Foulbrood (AFB) a disease of the honey bee larvae. In this study, the HE was partitioned into five main fractions by chromatographic techniques leading to the isolation of four known compounds: two prenylated phloroglucinol α-pyrones (1 and 3), 5,7-dihydroxy-3,8-dimethoxyflavone (gnaphaliin A) (2), and 23-methyl-6-O-demethylauricepyrone (4). Isolated compounds were further analyzed towards structural elucidation using 1H RMN and 13C RMN spectroscopic techniques. For the first time, the antimicrobial activity of the isolated compounds was evaluated against P. larvae strains by broth microdilution method and compared with that of the whole HE. Compounds 1-4 displayed minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values ranging between 0.07 and 62.5 µg/mL and 0.26 and 12.5 µg/mL, respectively. The lowest MIC and MBC values were obtained with compounds 3 and 4, respectively. The antimicrobial activity of each single compound and the combination of them showed that the presence of all compounds is needed for the antimicrobial efficacy of whole HE.