Biosynthesis of copper nanoparticles using Alstonia scholaris leaves and its antimicrobial studies.

The utilization of plants for the production of metallic nanoparticles is gaining significant attention in research. In this study, we conducted phytochemical screening of Alstonia scholaris (A. scholaris) leaves extracts using various solvents, including chloroform, ethyl acetate, n-hexane, methanol, and water. Our findings revealed higher proportions of flavonoids and alkaloids in both solvents compared to other phytochemical species. In the methanol, extract proteins, anthraquinone and reducing sugar were not detected. On the other hand, the aqueous extract demonstrated the presence of amino acids, reducing sugar, phenolic compounds, anthraquinone, and saponins. Notably, ethyl acetate and chloroform extracts displayed the highest levels of bioactive compounds among all solvents. Intrigued by these results, we proceeded to investigate the antibacterial properties of the leaf extracts against two major bacterial strains, Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). All extracts exhibited significant zones of inhibition against both bacterial isolates, with S. aureus showing higher susceptibility compared to E. coli. Notably, the methanol extract displayed the most potent I hibitory effect against all organisms. Inspired by the bioactivity of the methanol extract, we employed it as a plant-based material for the green synthesis of copper nanoparticles (Cu-NPs). The synthesized Cu-NPs were characterized using Fourier infrared spectroscopy (FT-IR), UV-visible spectroscopic analysis, and scanning electron microscopy (SEM). The observed color changes confirmed the successful formation of Cu-NPs, while the FTIR analysis matched previously reported peaks, further verifying the synthesis. The SEM micrographs indicated the irregular shapes of the surface particles. From the result obtained by energy dispersive X-ray spectroscopic analysis, Cu has the highest relative abundance of 67.41 wt%. Confirming the purity of the Cu-NPs colloid. These findings contribute to the growing field of eco-friendly nanotechnology and emphasize the significance of plant-mediated approaches in nanomaterial synthesis and biomedical applications.

Pro-apoptotic effect of chloroform fraction of Moraea sisyrinchium bulb against glioblastoma cells.

INTRODUCTION: Glioblastoma is a common malignant brain tumor, with limited therapeutic options. In our previous study, the Moraea sisyrinchium plant showed cytotoxicity against glioblastoma and hepatocellular carcinoma cells. Among different parts of this plant (flower, stem, and bulb), the bulb showed better anticancer potential. The present work aimed to test the anticancer activity of different fractions of the bulb extract, to determine its phytochemicals, and to study its mechanism action on glioblastoma. METHODS: The bulb extract was partitioned into different fractions using immiscible solvents. The U87 glioblastoma cells were incubated with the obtained fractions. Then, the cell proliferation assay (MTT), cell migration test (scratch), cell cycle analysis (propidium iodide staining), apoptosis/necrosis assay (annexin V/propidium iodide staining), and real-time PCR (PTEN, Akt, mTOR, BAX and BCL-2 genes) were performed. Phytochemicals were determined using liquid chromatography-mass spectroscopy. RESULTS: The chloroform fraction showed more antiproliferative effect than n-hexane, ethyl acetate, and n-butanol fractions. Also, chloroform fraction induced cell cycle arrest, increased apoptosis, and inhibited cell migration ability (P < 0.05). The expression of PTEN, mTOR, and BAX genes was significantly up-regulated, while the expression of Akt and Bcl-2 showed down-regulation. The phytochemicals identified in the chloroform fraction were mainly xanthones, phytosterols, and isoflavones. CONCLUSION: The chloroform fraction of Moraea sisyrinchium bulb inhibits the proliferation and migration of glioblastoma cells by inducing cell cycle arrest and apoptosis by upregulation of the PTEN gene and Bax/Bcl-2 ratio. The identified compounds in the chloroform fraction are potential candidates for further investigation as anticancer agents against glioblastoma.

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.

Alleviating Chlorinated Alkane Inhibition on Dehalococcoides to Achieve Detoxification of Chlorinated Aliphatic Cocontaminants.

Cocontamination by multiple chlorinated solvents is a prevalent issue in groundwater, presenting a formidable challenge for effective remediation. Despite the recognition of this issue, a comprehensive assessment of microbial detoxification processes involving chloroethenes and associated cocontaminants, along with the underpinning microbiome, remains absent. Moreover, strategies to mitigate the inhibitory effects of cocontaminants have not been reported. Here, we revealed that chloroform exhibited the most potent inhibitory effects, followed by 1,1,1-trichloroethane and 1,1,2-trichloroethane, on dechlorination of dichloroethenes (DCEs) in Dehalococcoides-containing consortia. The observed inhibition could be attributed to suppression of biosynthesis and enzymatic activity of reductive dehalogenases and growth of Dehalococcoides. Notably, cocontaminants more profoundly inhibited Dehalococcoides populations harboring the vcrA gene than those possessing the tceA gene, thereby explaining the accumulation of vinyl chloride under cocontaminant stress. Nonetheless, we successfully ameliorated cocontaminant inhibition by augmentation with Desulfitobacterium sp. strain PR owing to its ability to attenuate cocontaminants, resulting in concurrent detoxification of DCEs, trichloroethanes, and chloroform. Microbial community analyses demonstrated obvious alterations in taxonomic composition, structure, and assembly of the dechlorinating microbiome in the presence of cocontaminants, and introduction of strain PR reshaped the dechlorinating microbiome to be similar to its original state in the absence of cocontaminants. Altogether, these findings contribute to developing bioremediation technologies to clean up challenging sites polluted with multiple chlorinated solvents.

Antibacterial and antibiotic potentiating capabilities of extracts isolated from Burkillanthus malaccensis, Diospyros hasseltii and Cleisthanthus bracteosus against human pathogenic bacteria.

Antibiotics which once a boon in medicine and saved millions of lives are now facing an ever-growing menace of antibacterial resistance, which desperately needs new antibacterial drugs which are innovative in chemistry and mode of action. For many years, the world has turned to natural plants with antibacterial properties to combat antibiotic resistance. On that basis, we aimed to identify plants with antibacterial and antibiotic potentiating properties. Seventeen different extracts of 3 plants namely Burkillanthus malaccensis, Diospyros hasseltii and Cleisthanthus bracteosus were tested against multi-drug resistant Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Methicillinresistant Staphylococcus aureus (MRSA) and methicillin-susceptible Staphylococcus aureus (MSSA). Antibacterial activity of hexane, methanol and chloroform extracts of bark, seed, fruit, flesh and leaves from these plants were tested using, disk diffusion assay, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays. Antibiotic potentiating capabilities were tested using time-kill assay. B. malaccensis fruit chloroform extract showed the biggest zone of inhibition against MRSA (13.00±0.0 mm) but C. bracteosus bark methanol extract showed the biggest inhibition zone against MSSA (15.33±0.6 mm). Interestingly, bark methanol extract of C. bracteosus was active against MRSA (8.7±0.6 mm), MSSA (7.7±0.6 mm) (Gram-positive) and A. baumannii (7.7±0.6 mm) (Gram-negative). Overall, the leaf methanol and bark methanol extract of C. bracteosus warrants further investigation such as compound isolation and mechanism of action for validating its therapeutic use as antibiotic potentiator importantly against MRSA and A. baumannii.

Liver protective effect of chloroform extract of Bauhinia purpurea leaves is attributed partly to its antioxidant action and the presence of flavonoids.

CONTEXT: Bauhinia purpurea L. (Fabaceae) is used in the Ayurvedic system to treat various oxidative-related ailments (e.g., wounds, ulcers etc.). Therefore, it is believed that the plant also has the potential to alleviate oxidative-related liver damage. OBJECTIVE: This study elucidates the hepatoprotective activity of chloroform extract of B. purpurea leaves (CEBP) in paracetamol (PCM)-induced liver injury (PILI) rats. MATERIALS AND METHODS: Male Sprague-Dawley rats (n = 6) were pre-treated once daily (p.o.) with CEBP (50-500 mg/kg) for seven consecutive days before being administered (p.o.) a hepatotoxic agent, 3 g/kg PCM. Liver enzyme levels were determined from the collected blood, while the collected liver was used to determine the activity of endogenous antioxidant enzymes and for histopathological examination. CEBP was also subjected to radical scavenging assays and phytochemical analysis. RESULTS: CEBP significantly (p < 0.05) reversed the toxic effect of PCM by increasing the serum levels of AST and ALT, and the activity of endogenous catalase (CAT) and superoxide dismutase (SOD) while reducing the liver weight/body weight (LW/BW) ratio. Other than low TPC value and radical scavenging activity, CEBP had a high antioxidant capacity when evaluated using the oxygen radical absorbance capacity (ORAC) assay. UHPLC-ESI-MS analysis of CEBP showed the presence of flavonoids. DISCUSSION AND CONCLUSIONS: CEBP exerts its hepatoprotective activity through a non-free radical scavenging pathway that involves activation of the endogenous enzymatic antioxidant defense system. Further study is needed to identify the responsible bioactive compounds before the plant can be developed as a future alternative hepatoprotective medicament for clinical use.

Antireproductive changes instigated by efficient drug delivery via papaya seed chloroform extract-based nanoparticles in male rat Bandicota bengalensis.

Anti-reproductive potential of papaya seed chloroform extract-based solid lipid nanoparticles (PSCEN) was investigated for the first time in lesser bandicoot rat, Bandicota bengalensis. Mature male rats (n = 30 per group) were fed bait (loose mixture of cracked wheat, powdered sugar, and groundnut oil in the ratio 88:10:2) containing two different concentrations of PSCEN (5% and 10%) in a bi-choice condition for 15 days with one group as vehicle control. The ingestion of active ingredient in 15 days treatment was significantly (P ≤ 0.05) higher by rats treated with 10% PSCEN (39.17-58.70 g/kg body weight) as compared to rats treated with 5% PSCEN (21.30-33.23 g/kg body weight). A dose dependent significant (P ≤ 0.05) decrease was observed in the level of testosterone, FSH, LH and GnRH in plasma of treated rats. A significant (P ≤ 0.05) decrease was also observed in level of total soluble proteins, total lipids, phospholipids and cholesterol in both plasma and testicular tissue, and level of 17ß-HSD and 3ß-HSD in testicular tissue indicating anti-reproductive effects of PSCEN treatment. There was observed significant (P ≤ 0.05) effect of treatment on histomorphology of testis and cauda epididymis in the form of reduced tubular diameter, germinal epithelial thickness, number of germ cells and dissociation of epithelial cycle in seminiferous tubules, and reduced tubular diameter, increased epithelial thickness, vacuolization, loose contact of principle cells and reduced number of spermatozoa in the cauda epididymal tubules. Maximum antifertility effect was observed with 10% PSCEN treatment, which was not reversed upto 105 days of treatment withdrawal indicating long-term efficacy. The current investigation suggests the use of PSCEN in the management of reproduction of B. bengalensis by exerting influence on testicular and cauda epididymal functions and biochemical parameters.

Antibacterial Effects of Henna (Lawsonia inermis) Leaf Extracts (Chloroform) Against Two Food Borne and Nosocomial Infection Causing Pathogens: Staphylococcus aureus and Escherichia coli.

Evaluation of the in vitro antibacterial activity of Chloroform extracts isolated from Henna (Lawsonia inermis) leaf against two food born & nosocomial infection causing pathogens, gram-positive Staphylococcus aureus and gram-negative Escherichia coli. This interventional study was carried out during the period of January 2021 to December 2021 in the Department of Pharmacology and Therapeutics in collaboration with the Department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh. The antibacterial activity was tested at different concentrations of Chloroform Henna leaf extracts by using disc diffusion and broth dilution method. The extract was prepared by using solvents chloroform and 0.1% DMSO. The test microorganisms were also tested for their activity against a standard antibiotic Ciprofloxacin by broth dilution method and the result was compared with that of Chloroform extracts. Chloroform Henna Extracts (CHE) were used initially in nine different concentrations (2.5, 5, 10, 20, 50, 100, 200, 500 and 1000mg/ml) and later in selected concentrations as needed to confirm the more precise margin of antimicrobial sensitivity of the extracts. Among different concentrations of the CHE, 100mg/ml and above concentrations showed inhibitory effect against Staphylococcus aureus and 300mg/ml and above concentrations showed inhibitory effect against Escherichia coli. The MIC for Staphylococcus aureus and Escherichia coli were 100 and 350mg/ml in CHE respectively. The MIC of Ciprofloxacin was 1µg/ml against both Staphylococcus aureus and Escherichia coli. The MIC of Ciprofloxacin was the lowest in comparison to MICs of CHE for the test organisms. The present study showed that Chloroform Henna extracts demonstrated antibacterial effects against food borne pathogens. From this study, it is clearly observed that there is definite antibacterial effect of the chloroform extract of Henna leaves against Staphylococcus aureus and Escherichia coli.

Chloroform extracts of Atractylodes chinensis inhibit the adhesion and invasion of Salmonella typhimurium.

There are 27 million cases of Salmonella Typhimurium (STM) reported worldwide annually, which have resulted in 217,000 deaths to date. Thus, there is an urgent requirement to develop novel antibacterial agents to target the multidrug-resistant strains of STM. We evaluated the inhibitory effect of the chloroform extracts of Atractylodes chinensis (Ac-CE) on the virulence of STM in vitro and develop it as a potential antibacterial agent. First, we determined the in vitro effects of Ac-CE on STM biofilm formation, and swimming, swarming, and adhesion to mucin. Further, we evaluated the effect of Ac-CE on the adhesion and invasion of STM at the gene level. Lastly, we evaluated the inhibitory effect of Ac-CE on STM infectivity at the cellular level. Ac-CE could attenuate both the adhesion and invasion abilities of STM in vitro. At the gene level, it could inhibit the expression of flagella, pilus, biofilm, SPI-1, and SPI-2 genes, which are related to the adhesion and invasion ability of STM in cells. Ac-CE significantly downregulated the expression of inflammatory cytokines and the TLR4/MyD88/NF-κB pathway in an STM infection cell model. It also significantly recovered the expression of intestinal barrier-related genes and proteins in intestinal cells that are damaged during STM infection. Ac-CE is effective as an antivirulence agent in alleviating STM infection. Although the main components of Ac-CE were analyzed.We have not demonstrated the antivirulence effect of the active ingredients in Ac-CE. And the antivirulence effect of Ac-CE and its active ingredients warrant further in vivo studies.

Antiapoptotic and antioxidative efficacy of rhizomes of Curcuma amada on the management of diabetes-induced male infertility in albino rat: An effective fraction selection study.

Men with diabetes have negative effects on reproduction that causes sexual dysfunction. Medicinal plants are non-toxic and much safer than synthetic drugs because regular use of synthetic drugs shows long-term side effects. Curcuma amada (Roxb) is a medicinal plant used in Ayurveda and Unani medicinal systems in India. The goal of this study is to rummage the potential efficiency of the most potent solvent fraction of effective extract of hydro-methanol 60:40 of C. amada rhizome on male gonadal hypofunction in streptozotocin-induced diabetic rat. Diabetes-induced testicular hypofunction was evaluated by glycemic, spermiological, biochemical, genomic, flow cytometric, and histology of testicular tissue. The n-hexane, chloroform, ethyl-acetate, and n-butanol solvent fractions of the said extract were administrated for 4 weeks at 10 mg dose/100 g body weight/day. Among all the used fractions, the ethyl-acetate solvent fraction-treated group showed maximum recovery in serum insulin (177.42%), sperm count (92.84%), sperm motility (97.15%), and serum testosterone (164.33%). The diabetic rats treated with ethyl-acetate solvent fraction also exhibited the maximum resettlement in flow cytometric analysis of sperm viability (55.84%) and sperm mitochondrial integrity (149.79%), gene expression patterns of key markers for androgenesis (Δ5, 3ß-HSD 87.50%, and 17ß-HSD 74.66%) and apoptosis (Bax 44.63%, Bcl-2 54.03%, and Caspase-3 35.77%) along with testicular histology. The ethyl-acetate fraction contains alkaloids, flavonoids, and polyphenols where all of these components are not present in other fractions, may be the most effective cause for the recovery of diabetes-linked oxidative stress-mediated testicular hypofunctions. PRACTICAL APPLICATIONS: Nowadays worldwide, the use of synthetic drugs are reduced due to their toxic effect. At present, synthetic drugs are replaced by several herbal drugs, the natural source of medicine which has many therapeutic values. C. amada has strong antioxidant activity due to the presence of bio-active compound(s) that can able to manage streptozotocin-induced diabetes linked to oxidative damage of male gonadal organs. Therefore, these bio-active compound(s)-containing said medicinal plant may use as a good source of antioxidative food in the food industry as nutraceuticals and in pharmaceutical industries for the development of the herbal drug to manage diabetes-linked male gonadal hypofunctions. At present, WHO also gives emphasis for developing one drug-multi-disease therapy. From such a viewpoint, this active fraction-containing phytomolecules may have corrective efficacy against diabetes as well as oxidative stress-linked testicular complications.

[Investigation of the Anti-Leishmanial Effects of Prangos ferulacea and Ferula orientalis Extracts Collected from Sirnak Province Against Leishmania tropica Isolated in Turkey]. / Sirnak Ili ve Çevresinden Toplanan Prangos ferulacea ve Ferula orientalis Ekstrelerinin Türkiye'den Izole Edilmis Leishmania tropica'ya Karsi Anti-Leishmanial Etkilerinin Arastirilmasi.

Leishmaniasis is a vector-borne disease that is caused by the protozoa of Leishmania genus. Leishmaniasis is endemic in tropical, subtropical, and large areas of the Mediterranean basin, and covers a total of 98 countries worldwide. It is estimated, according to the World Health Organization (WHO) data, that approximately 350 million people are at risk in these areas, and approximately 12 million people are infected. Increased drug resistance has been documented lately, in the treatment of leishmaniasis which causes almost 1.2 million new cases annually. Thus, interest in plant-derived active substances has increased in recent years, and new anti-leishmanial agents are investigated with in vitro studies. The aim of the present study was to investigate the anti-leishmanial effects of Prangos ferulacea and Ferula orientalis plant extracts collected from the rural areas of Sirnak province against Leishmania tropica. The water, chloroform, and ethanol extracts of the roots, stems, and fruits of P.ferulaceae and F.orientalis plants were obtained, and the cytotoxic activity tests of the extracts were performed. L.tropica isolate obtained from the Parasite Bank in Manisa Celal Bayar University in Turkey (MHOM/TR/2012/CBCL-LT) was grown on NNN and RPMI 1640 broth medium. The cytotoxicity of each extract on the L.tropica isolate was evaluated with the XTT test. Amphotericin B (AmpB) was used as the positive control, and the IC50 values were determined. The lowest IC50 values of the plant extracts were found to be as follows: P.ferulaceae root chloroform extract 36 µg/ml and fruit chloroform extract 20 µg/ml, F.orientalis root ethanol extract 2.5 µg/ml, and fruit ethanol extract 48 µg/ml, stem chloroform extract 24 µg/ml, and fruit chloroform extract 3.1 µg/ml. It was also determined in our study that only P.ferulaceae root ethanol extract showed cytotoxic activity on the WI-38 fetal lung fibroblast cell line at 65.19 µg/ml at 72 hours. This is the first study that assessed the anti-leishmanial activities of P.ferulaceae and F.orientalis plants that grow in high altitude areas of our country. It was determined that P.ferulaceae root ethanol extract and fruit chloroform extract had the lowest IC50 values among the 18 plant extracts that we examined for their anti-leishmanial activities. The outcomes of this study will be useful in further studies for the determination of active compounds in P.ferulaceae and F.orientalis plant extracts.

Fungi anaesthesia.

Electrical activity of fungus Pleurotus ostreatus is characterised by slow (h) irregular waves of baseline potential drift and fast (min) action potential likes spikes of the electrical potential. An exposure of the myceliated substrate to a chloroform vapour lead to several fold decrease of the baseline potential waves and increase of their duration. The chloroform vapour also causes either complete cessation of spiking activity or substantial reduction of the spiking frequency. Removal of the chloroform vapour from the growth containers leads to a gradual restoration of the mycelium electrical activity.

Phytochemical Investigation and Therapeutical Potential of Cotinus coggygria Scop. in Alloxan-Induced Diabetic Mice.

Plants are a significant source for the development of new phytomedicines due to their great clinical benefits, efficiency, cost-effectiveness, fewer side effects, and more affordable therapies. Numerous plants used in traditional treatments, such as Cotinus coggygria Scop., have been effective in the treatment of diabetes mellitus (DM). Therefore, the study is aimed at assessing the phytochemical, antioxidant, and antidiabetic properties of C. coggygria. The hypoglycemic and hypolipidemic activity was evaluated in Swiss male Albino mice by administering an oral dose of 150-250 mg/kg of C. coggygria extracts in alloxan-induced diabetic mice for 15 days. The antioxidant activity and phytochemical composition of the extracts were assessed by using α, α diphenyl-ß-picrylhydrazyl (DPPH) and hydrogen peroxide scavenging assays and through standard chemical procedures. The effects of extracts on blood glucose, body weight, lipid profile, and biochemical parameters like total cholesterol (TC), triglyceride (TG), low-density lipids (LDL), high-density lipids (HDL), plasma insulin, liver glycogen, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), urea, and creatinine were determined according to standard procedures. The activities of antioxidant enzymes such as superoxide-dismutase (SOD), peroxidase (POD), and catalase (CAT) were also analyzed spectrophotometrically. The hypoglycemic and hypolipidemic effects with chloroform extracts of 250 mg/kg were found significant in the treatment of diabetes in alloxanised mice compared to the diabetic group. The haematological parameters such as TC, TG, HDL, LDL, creatinine, urea, AST, ALT, and ALP were significantly improved (p < 0.01) by the chloroform extract of 250 mg/kg compared to the diabetic group. Treatment for 15 days showed significant elevation (p < 0.01) of antioxidant enzymes. Fourier-transform infrared spectroscopic (FTIR) and gas chromatography-mass spectrometry (GC-MS), column chromatography (CC), and nuclear magnetic resonance (NMR) analyses tentatively identified different phytoconstitutents and metabolites in C. coggygria leaves, which have been reported to possess antihyperglycemic properties. In conclusion, the chloroform extract of 250 mg/kg of C. coggygria possesses significant hypoglycemic and hypolipidemic potential which may prove the claimed use of the plant in amelioration of diabetes and associated complications in folkloric medicine. Additional studies are required for the purification, characterization, and structural elucidation of bioactive compounds.

Cytotoxic activity of the chloroform fraction of Piper aduncum and its effect on the cell cycle in gastric cancer cell lines. / Actividad citotóxica de la fracción clorofórmica de Piper aduncum y su efecto en el ciclo celular en líneas celulares de cáncer gástrico.

OBJECTIVES: To evaluate the cytotoxic activity of the chloroform fraction of the Piper aduncum methanolic extract (PAMoCl) and its effect on the cell cycle in two gastric cancer cell lines: AGS and KATO III. MATERIALS AND METHODS: The cytotoxic effect of PAMoCl was evaluated in cell lines AGS and KATO III. The following PAMoCl concentrations were tested, 1.25, 2.5, 5, 10, 20, 40, 80 and 160 µg/mL. Resazurine was used to evaluate cell viability. In the cell cycle assay, the cells were treated with 19.62 µg/mL and 39.23 µg/mL of PAMoCl for AGS as well as 87.49 µg/mL and 160 µg/mL for KATO III. After 24 hours both cell lines were analyzed by flow cytometry. RESULTS: PAMoCl showed cytotoxic activity, inhibiting cell growth by 50%. It presented a (IC50) of 39.23 µg/mL and 87.49 µg/mL at 24 hours and a (IC50) of 49.47 µg/mL and 64.68 µg/mL at 48 hours against AGS and KATO III cell lines, respectively. In addition, it was observed that PAMoCl has an effect on the cell cycle, it causes an accumulation of cells in the G2/M phase. CONCLUSIONS: PAMoCl contains secondary metabolites with cytotoxic activity that have an effect on the G2/M phase of the cell cycle, in two gastric cancer cell lines, both primary and metastatic. The results of this study will allow us to deepen the search for more effective active ingredients found in PAMoCl for eliminating gastric cancer cells, but with less toxicity for healthy cells.

OBJETIVOS: Evaluar la actividad citotóxica de la fracción clorofórmica del extracto metanólico de Piper aduncum (PAMoCl) y su efecto en el ciclo celular en dos líneas celulares de cáncer gástrico: AGS y KATO III. MATERIALES Y MÉTODOS: El efecto citotóxico de PAMoCl se evaluó en las líneas celulares: AGS y KATO III. Se probaron concentraciones de PAMoCl: 1,25; 2,5; 5; 10; 20; 40; 80 y 160 µg/mL. Para evaluar la viabilidad celular se usó el reactivo resazurina. En el ensayo de ciclo celular las células fueron tratadas con 19,62 µg/mL y 39,23 µg/mL de PAMoCl para AGS, así como 87,49 µg/mL y 160 µg/mL para KATO III. Después de 24 horas ambas líneas celulares fueron analizadas por citometría de flujo. RESULTADOS: PAMoCl mostró actividad citotóxica con una inhibición del crecimiento celular en un 50% (IC50) de 39,23 µg/mL y 87,49 µg/mL a las 24 horas y un IC50 de 49,47 µg/mL y 64,68 µg/mL a las 48 horas frente a las líneas celulares AGS y KATO III, respectivamente. Además, se observó que PAMoCl tiene efecto a nivel del ciclo celular: provoca una acumulación de células en la fase G2/M. CONCLUSIONES: PAMoCl contiene metabolitos secundarios con actividad citotóxica que tienen efecto en la fase G2/M del ciclo celular, en dos líneas celulares de cáncer gástrico tanto primario como metastásico. Los resultados de este estudio permitirán profundizar en la búsqueda de principios activos presentes en PAMoCl que tengan mayor eficacia en la eliminación de células de cáncer gástrico, pero con menor toxicidad en células sanas.

Bioprospecting Staphylococcus Phages with Therapeutic and Bio-Control Potential.

Emergence of antibiotic-resistant bacteria is a serious threat to the public health. This is also true for Staphylococcus aureus and other staphylococci. Staphylococcus phages Stab20, Stab21, Stab22, and Stab23, were isolated in Albania. Based on genomic and phylogenetic analysis, they were classified to genus Kayvirus of the subfamily Twortvirinae. In this work, we describe the in-depth characterization of the phages that electron microscopy confirmed to be myoviruses. These phages showed tolerance to pH range of 5.4 to 9.4, to maximum UV radiation energy of 25 µJ/cm2, to temperatures up to 45 °C, and to ethanol concentrations up to 25%, and complete resistance to chloroform. The adsorption rate constants of the phages ranged between 1.0 × 10-9 mL/min and 4.7 × 10-9 mL/min, and the burst size was from 42 to 130 plaque-forming units. The phages Stab20, 21, 22, and 23, originally isolated using Staphylococcusxylosus as a host, demonstrated varied host ranges among different Staphylococcus strains suggesting that they could be included in cocktail formulations for therapeutic or bio-control purpose. Phage particle proteomes, consisting on average of ca 60-70 gene products, revealed, in addition to straight-forward structural proteins, also the presence of enzymes such DNA polymerase, helicases, recombinases, exonucleases, and RNA ligase polymer. They are likely to be injected into the bacteria along with the genomic DNA to take over the host metabolism as soon as possible after infection.

Hemistepsin A alleviates liver fibrosis by inducing apoptosis of activated hepatic stellate cells via inhibition of nuclear factor-κB and Akt.

Hemistepsin A (HsA), isolated from Hemistepta lyrata (Bunge) Bunge, has the ability to ameliorate hepatitis in mice. However, the effects of H. lyrata and HsA on other types of liver disease have not been explored. In this report, we investigated the effects of H. lyrata and HsA on liver fibrosis and the underlying molecular mechanisms in activated hepatic stellate cells (HSCs). Based on cell viability-guided isolation, we found HsA was the major natural product responsible for H. lyrata-mediated cytotoxicity in LX-2 cells. HsA significantly decreased the viability of LX-2 cells and primary activated HSCs, increased the binding of Annexin V, and altered the expression of apoptosis-related proteins, suggesting that HsA induces apoptosis in activated HSCs. HsA reduced the phosphorylation of IKKε and the transactivation of nuclear factor-κB (NF-κB). Moreover, HsA decreased the phosphorylation of Akt and its downstream signaling molecules. Transfection experiments suggested that inhibition of NF-κB or Akt is essential for HsA-induced apoptosis of HSCs. In a CCl4-induced liver fibrosis model, HsA administration significantly decreased ALT and AST activities. Furthermore, HsA attenuated CCl4-mediated collagen deposits and profibrogenic genes expression in hepatic tissue. Thus, HsA may serve as a natural product for managing liver fibrosis through inhibition of NF-κB/Akt-dependent signaling.

Protective Effect of Spirulina platensis Extract against Dextran-Sulfate-Sodium-Induced Ulcerative Colitis in Rats.

Inflammatory bowel disease is a multifactorial inflammatory condition. This study aimed to test the protective effects of Spirulina platensis against ulcerative colitis (UC). UC was induced in thirty-six male Wistar rats by adding dextran sulfate sodium (DSS) to their drinking water, while a control group received only drinking water. UC rats were equally-divided into six groups that received a single oral daily dose of vehicle (DSS), sulfasalazine (SSZ, 50 mg/kg/day), chloroform or the hydroalcoholic extracts of Spirulina platensis (100 or 200 mg/kg/day) for 15 days, and then blood and colon samples were harvested for determination of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), erythrocyte sedimentation rate (ESR), myeloperoxidase (MPO), and histopathology. At the end of the study, compared to time-matched controls, UC rats showed increased TNF-α (1.64-fold), IL-6 (5.73-fold), ESR (3.18-fold), and MPO (1.61-fold), along with loss of body weight (24.73%) and disease activity index (1.767 ± 0.216 vs. 0 ± 0), p < 0.001. These effects were prevented by SSZ treatment (p < 0.001 vs. DSS). The hydroalcoholic extract of Spirulina platensis dose-dependently modulated all DSS-induced inflammatory changes. However, the chloroform extract significantly lowered only IL-6 and ESR, but not TNF-α or MPO levels. The protective effects of the hydroalcoholic extract of Spirulina platensis against experimental UC involved mitigation of DSS-induced inflammation.

Proposition of a safe Mycobacterium tuberculosis complex denaturation method that does not compromise the integrity of DNA for whole-genome sequencing.

Whole-genome sequencing plays now a leading role in epidemiologic studies of tuberculosis. DNA extraction of Mycobacterium tuberculosis complex (MTBC) requires complete inactivation of the strains, to be handled for further molecular procedures. In this study we compared two chloroform-based denaturation methods (one with a step of heat killing, one without) to a traditional heat inactivation method. Our results showed that 40% of the strains of MTBC treated by the traditional protocol resulted in a positive culture whereas no culture was observed with the two chloroform-based protocols. The DNA extracts obtained with chloroform-based protocols preparation were successfully used for whole-genome sequencing. We recommend inactivation with our rapid and efficient denaturation method using chloroform without heat killing which met our expectations and biosecurity requirements.

A general dose-response relationship for chronic chemical and other health stressors and mixtures based on an emergent illness severity model.

Current efforts to assess human health response to chemicals based on high-throughput in vitro assay data on intra-cellular changes have been hindered for some illnesses by lack of information on higher-level extracellular, inter-organ, and organism-level interactions. However, a dose-response function (DRF), informed by various levels of information including apical health response, can represent a template for convergent top-down, bottom-up analysis. In this paper, a general DRF for chronic chemical and other health stressors and mixtures is derived based on a general first-order model previously derived and demonstrated for illness progression. The derivation accounts for essential autocorrelation among initiating event magnitudes along a toxicological mode of action, typical of complex processes in general, and reveals the inverse relationship between the minimum illness-inducing dose, and the illness severity per unit dose (both variable across a population). The resulting emergent DRF is theoretically scale-inclusive and amenable to low-dose extrapolation. The two-parameter single-toxicant version can be monotonic or sigmoidal, and is demonstrated preferable to traditional models (multistage, lognormal, generalized linear) for the published cancer and non-cancer datasets analyzed: chloroform (induced liver necrosis in female mice); bromate (induced dysplastic focia in male inbred rats); and 2-acetylaminofluorene (induced liver neoplasms and bladder carcinomas in 20,328 female mice). Common- and dissimilar-mode mixture models are demonstrated versus orthogonal data on toluene/benzene mixtures (mortality in Japanese medaka, Oryzias latipes, following embryonic exposure). Findings support previous empirical demonstration, and also reveal how a chemical with a typical monotonically-increasing DRF can display a J-shaped DRF when a second, antagonistic common-mode chemical is present. Overall, the general DRF derived here based on an autocorrelated first-order model appears to provide both a strong theoretical/biological basis for, as well as an accurate statistical description of, a diverse, albeit small, sample of observed dose-response data. The further generalizability of this conclusion can be tested in future analyses comparing with traditional modeling approaches across a broader range of datasets.

Antioxidant and Antimutagenic Activities of Different Fractions from the Leaves of Rhododendron arboreum Sm. and Their GC-MS Profiling.

In this era of urbanization and environmental pollution, antioxidants and antimutagens derived from plants are promising safeguards for human health. In the current investigation, we analyzed the antioxidant and antimutagenic effects of the hexane, chloroform, and ethyl acetate fractions of Rhododendron arboreum Sm. leaves and determined their chemical composition. The different fractions inhibited lipid peroxidation, repressed the production of nitric oxide radicals, and prevented deoxyribose degradation. The antimutagenic activity of the leaf fractions was analyzed against 4-nitro-O-phenylenediamine, sodium azide and 2-aminofluorene mutagens in two test strains (TA-98 and TA-100) of Salmonella typhimurium. The experiment was conducted using pre- and co-incubation modes. The best results were obtained in the pre-incubation mode, and against indirect acting mutagen. The presence of a number of bioactive constituents was confirmed in the different fractions by GC-MS analysis. The study reveals the strong antioxidant and antimutagenic activity of R. arboreum leaves. We propose that those activities of R. arboreum might correspond to the combined effect of the phytochemicals identified by GC-MS analysis. To the best of our knowledge, this is the first report on the antimutagenic activity of R. arboreum leaves.