Chloroform Treatment Research Articles

Seasonal variation in characteristics of wear microparticles of high density (> 1.8 g cm−3) produced on road

Wear microparticles are produced on roads by traffic, and they can be transferred to rivers and seas settling as sediments. The sedimentation rate increases with increasing particle density and size. In this study, the types and amounts of high-density wear microparticles (HDWPs, >1.8 g cm−3) in road dust were investigated. The HDWPs ranging from 106 to 1000 μm were classified into eight categories depending on the color, shape, and physical property: mineral particles (MPs), asphalt pavement wear particles (APWPs), glass particles (GPs), glass beads (GBs), tire-road wear particles (TRWPs), plant-related particles (PRPs), road paint wear particles (RPWPs), and plastic particles (PPs). The HDWPs in road dust were the most abundant in winter (94.0–95.6 wt%), while being the lowest in spring (82.7–90.7 wt%). MPs accounted for over 50 wt% of the HDWPs; however, TRWPs were not found in HDWPs larger than 200 μm. The HDWPs produced by the abrasion of roads, including asphalt pavements and marking paint, exceeded 90 wt%. The non-crosslinked organic components in the HDWPs were removed by chloroform treatment. The chloroform-soluble components in the HDWPs were much more present in winter than in other seasons. Swelling TRWPs with chloroform released mineral particles on the surface.

Comparison of phytochemical composition, antimicrobial, antifungal, and antioxidant activities of lipophilic and ethanolic green tea leaf extracts

Infection diseases are a worldwide important problem for medicine and pharmacy. Today, there is a high interest in the evolution of new antimicrobial drugs due to the increasing percentage of resistant bacteria and fungi strains. Green tea leaf contains a variety of natural compounds that are possible to apply in the creation of new antimicrobial drugs. The aim of the work was to compare phytochemical composition, antimicrobial, antifungal activities of lipophilic and ethanolic green tea leaf extracts. Materials and methods. The object of the research was the lipophilic extract obtained using chloroform, along with two ethanolic extracts of green tea leaves. One of the ethanolic extracts had been previously processed with chloroform, while the other had not. Antioxidant activity was determined by the potentiometric method, while antimicrobial and antifungal activities were assessed by the “wells” method. Results. The lipophilic extract of green tea leaves contains predominantly caffeine and organic acids, with the lowest amount of phenolic compounds. In contrast, the ethanolic extracts show the opposite pattern, with phenolic compounds predominating and caffeine present in lower quantities. The lipophilic extract exhibits greater inhibition of the growth of S. aureus, E. coli, P. vulgaris, B. subtilis, and C. albicans compared to the 96 % ethanolic extract processed with chloroform, with inhibition rates of 19 %, 18 %, 12 %, 12 %, 16 %, and 20 % respectively. When comparing antimicrobial activity to the 96 % ethanolic extract without chloroform treatment, the results remained consistent. The antioxidant activity of the lipophilic extract was 58.7 and 60.0 times lower than that of the 96 % ethanolic extract processed with chloroform and the 96 % ethanolic extract without treatment, respectively. Conclusions. The study revealed that the lipophilic extract exhibited greater inhibition of the growth of both Gram-positive and Gram-negative bacteria as well as fungi compared to the ethanolic extracts. However, it showed a lower level of antioxidant activity. It is hypothesized that caffeine, organic acids, and catechins may interact synergistically to enhance the antimicrobial and antifungal activity of green tea leaf extracts. The lipophilic extract shows promise for further development in the production of antimicrobial and antifungal drugs.

A Method for Epicuticular Wax and Contaminant Removal from Herbarium Specimens for Scanning Electron Microscopy

Abstract We describe a method for cleaning leaf epicuticular wax and contaminants from herbarium specimens in preparation for scanning electron microscopy. Specimens are rinsed in chloroform, heated at 50°C for 8 hrs, ultrasonicated in 70% ethanol, and subjected to critical-point drying. This method helps preserve and reveal the surface details of specimens that are heavily covered with epicuticular wax and contaminants, making many morphologically and functionally important details readily visible. We tested our method by using it to clean herbarium specimens and comparing the results with those obtained using three other chemicals: acetone, xylene, and 1:1 acetone:xylene, confirming that chloroform treatment was the most effective.

Fabrication and characterization of graphene coated nickel electrodes with internally stacked double layer supercapacitors

Supercapacitors are getting a lot of attention as high-performance energy storage devices that can find applications in multiple fields including high-power military devices and wearable or portable electronics. For a better supercapacitor device electrode plays a vital role and graphene is being widely studied as an efficient material for electrodes due to its large specific surface area of 2630 m2/g. However, the disordered alignment and the severe aggregation hinder the maximum utilization of its surface area. In this work nickel nitrate and chloroform treatments were used to improve the deposition of graphene on the current collector and to improve the substrate surface quality, respectively. In addition, the stacking technique is used to improve the overall energy storage capability of the device. The electrophoretic deposition (EPD) coating method was used to deposit graphene on the current collector. The important optimization was also performed in a KOH-based 3-electrode system and the chloroform treated and graphene-coated electrode showed an improved capacitance of 151 F/g at scan rate 0.01 V/sec as compared to the 126 F/g of the untreated sample. The graphene-coated electrode showed good cyclic stability with capacitance fading of ∼4.4 × 10−6%/cycle and a higher operating voltage up to 6 V has been achieved by utilizing the stacking method.

Isolation and therapeutic efficacy of enteropathogenic E. coli phage cocktail against colibacillosis in neonatal goats

Phage based therapeutics have shown promising results against the infections caused by the drug resistant bacteria. To combat the problem of antibiotic resistance posed by diarrhoeagenic E. coli, here, we identified and characterized 38 E. coli phages which were isolated from 70 solid sources (goat-faeces and soil). The in vitro lytic range of phage isolates (n=38) against 439 isolates of E. coli was found between 16 and 53%. Three phage isolates with highest host range showed lytic efficacy against 53, 48 and 46% of E. coli isolates, respectively. A preparation with above three phages was developed, and the phages of the preparation were found stable at wide range of temperature, pH and chloroform treatment. Endotoxin content of the preparation was found below the threshold level and it also passed safety and sterility tests. a total of 40 diarrheic goat kids were administered orally with the therapeutic phage preparation for two days twice daily. Total 21 diarrheic goat-kids were successfully treated using the therapeutic phage preparation, whereas 19 kids could not be treated (success rate: 52.5%; 21/40). The results of the current study provide insight for using lytic bacteriophages for therapeutic interventions against drug resistant E. coli responsible for colibacillosis in neonatal goat kids

Influence of Surface Chemistry of Carbon Nanofibers on the Hydrodechlorination of Chloroform to Olefins

Functionalized carbon nanofibers (CNF) are fascinating materials to be used as supports in Pd-based catalysts for the treatment of waste chloroform (TCM) to produce light olefins through the catalytic hydrodechlorination (HDC). The CNF were functionalized by HNO3, HCl, and urea. Compared to the Pd supported on un-treated CNF, all the catalysts using functionalized CNF as support showed lower turnover frequency values with higher stability, owing to their smaller Pd nanoparticles (NPs). These smaller Pd NPs are formed due to the stronger metal–support interactions promoted by the higher concentration of surface groups on the functionalized catalysts. Since the smaller Pd NPs could hinder the hydrogenation of olefins to paraffins, the selectivity to olefins increased on the functionalized catalysts. Moreover, the N-doped CNF was successfully formed on the catalyst functionalized by urea. Since the nitrogen functional groups (pyridinic N and pyrrolic N) could provide much stronger metal–support interactions compared to the oxygen functional groups on the other catalysts, the catalyst functionalized by urea showed the smallest Pd NPs among the four catalysts, leading to the highest selectivity to light olefins.

Disentangled ultrahigh molecular weight polyethylene thin film as a transparent substrate for flexible flat panel display

AbstractThe use of polymeric thin film as a substrate material in fabrication of flexible organic light‐emitting diodes (OLEDs) devices is well known. Polyethylene terephthalate (PET) and polyethylene naphtalate (PEN) thin films are used for such purpose commercially. In the present work, 10g of disentangled ultrahigh molecular weight polyethylene (D‐UHMWPE) was processed into 35 ± 5μ thin film by continuous solid state compaction process between two heated rollers of roll mill. The temperature of both rollers was kept at 125°C as well as 128°C and the thin‐film production rate was 1.5 m/min. Subsequently film was studied for substrate application in fabrication of flexible OLEDs. The primary requirement of OLED substrate material namely chemical stability, transparency, and dimensional stability of D‐UHMWPE thin film were studied. D‐UHMWPE film showed excellent chemical stability under various solvents (chloroform, chlorobenzene, dichlorobenzene, acetone, and isopropanol) treatments along with >80% transparency and dimensional stability up to 100 ± 10°C (machine and transverse directions 0.001% and 0% at 90°C and 0.003% and 0% at 110°C). Finally, OLED device was fabricated on D‐UHMWPE surface which showed excellent performance in terms of (light emitting) even when the devices were bended.

Factors Involved in Removing the Non-Structural Protein of Foot-and-Mouth Disease Virus by Chloroform and Scale-Up Production of High-Purity Vaccine Antigens.

Foot-and-mouth disease (FMD) is an economically important and highly infectious viral disease, predominantly controlled by vaccination. The removal of non-structural proteins (NSPs) is very important in the process of FMD vaccine production, because vaccinated and naturally infected animals can be distinguished by the presence of NSP antibodies in the FMD serological surveillance. A previous study reported that 3AB protein, a representative of NSPs, was removed by chloroform treatment. Therefore, in this study, the causes of 3AB removal and factors affecting the effect of chloroform were investigated. As a result, the effectiveness of chloroform differed depending on the virus production medium and was eliminated by detergents. In addition, it was found that 3AB protein removal by chloroform is due to the transmembrane domain of the N-terminal region (59–76 amino acid domain). Further, industrial applicability was verified by applying the chloroform treatment process to scale-up FMD vaccine antigen production. A novel downstream process using ultrafiltration instead of polyethylene glycol precipitation for high-purity FMD vaccine antigen production was established. This result will contribute toward simplifying the conventional process of manufacturing FMD vaccine antigens and ultimately reducing the time and cost of vaccine production.

Evaluation of Pre-Analytical and Analytical Methods for Detecting SARS-CoV-2 in Municipal Wastewater Samples in Northern Italy

(1) Background: The surveillance of SARS-CoV-2 RNA in urban wastewaters allows one to monitor the presence of the virus in a population, including asymptomatic and symptomatic individuals, capturing the real circulation of this pathogen. The aim of this study was to evaluate the performance of different pre-analytical and analytical methods for identifying the presence of SARS-CoV-2 in untreated municipal wastewaters samples by conducting an inter-laboratory proficiency test. (2) Methods: three methods of concentration, namely, (A) Dextran and PEG-6000 two-phase separation, (B) PEG-8000 precipitation without a chloroform purification step and (C) PEG-8000 precipitation with a chloroform purification step were combined with three different protocols of RNA extraction by using commercial kits and were tested by using two primers/probe sets in three different master mixes. (3) Results: PEG-8000 precipitation without chloroform treatment showed the best performance in the SARS-CoV-2 recovery; no major differences were observed among the protocol of RNA extraction and the one-step real-time RT-PCR master mix kits. The highest analytic sensitivity was observed by using primers/probe sets targeting the N1/N3 fragments of SARS-CoV-2. (4) Conclusions: PEG-8000 precipitation in combination with real-time RT-PCR targeting the N gene (two fragments) was the best performing workflow for the detection of SARS-CoV-2 RNA in municipal wastewaters.

Isolation and Characterization of a Novel Lytic Halotolerant Phage from Yuncheng Saline Lake.

Halophilic phage are a type of virus that exist in salty environments within halophilic archaeal or bacterial hosts. However, relatively few reports on halophilic bacteriophages exist, and our overall understanding of halophilic bacteriophages is quite limited. We used SYBR Green I fluorescent staining to detect the abundance of viruses in Yuncheng Saline Lake, China. Using the double-layer plate method, a lytic phage that could infect halophilic bacterium Salinivibrio sp. YM-43 was isolated and named YXM43. We studied host range, optimal host, morphological characteristics, nucleic acid type, protein composition, and other biological characteristics of the virus. Results reveal a high abundance of this halophilic virus in Yuncheng Saline Lake. The newly isolated bacteriophage YXM43 has a narrow host range, with the most suitable host being Virgibacillus sp. SK39. After purification and enrichment, YXM43 is observed as a spherical particle with a diameter of approximately 30nm, with no tail. No lipid envelope can be seen in YXM43. The capsid protein of the virus can be separated into seven proteins with molecular weights ranging from 62.0 to 13.0kDa. YXM43 is a DNA virus with a genome approximately 23kb. The virus is tolerant of low salinity, and its activity is highest at a temperature of 60°C and a pH of 10. YXM43 is temperature and pH tolerant, and can adapt to environmental change, even withstanding chloroform treatment. The results indicate that bacteriophage YXM43 is a novel halophilic bacteriophage with broad tolerance to environmental change.

An improved RT-qPCR method for direct quantification of enveloped RNA viruses

Reverse transcription quantitative PCR (RT-qPCR) has emerged as the gold standard for virus detection and quantification, being utilized in numerous diagnostic and research applications. However, the direct detection of viruses has so far posed a challenge as the viral genome is often encapsidated by a proteinaceous layer surrounded by a lipid envelope. This necessitates an additional and undesired RNA extraction step prior to RT-qPCR amplification. To circumvent this limitation, we have developed a direct RT-qPCR method for the detection of RNA viruses. In our method, we provide a proof-of-concept using phage phi6, a safe-to-use proxy for pathogenic enveloped RNA viruses that is commonly utilized in e.g. aerosolization studies. First, the phage phi6 envelope is removed by 1% chloroform treatment and the virus is then directly quantified by RT-qPCR. To identify false negative results, firefly luciferase is included as a synthetic external control. Thanks to the duplex format, our direct RT-qPCR method reduces the reagents needed and provides an easy to implement and broadly applicable, fast, and cost-effective tool for the quantitative analysis of enveloped RNA viruses.•One-step direct RT-qPCR quantification of phage phi6 virus without prior RNA isolation.•Reduced reaction volume for sustainable and cost-effective analysis.

Isolation and Identification of a Wastewater Siphoviridae Bacteriophage Targeting Multidrug-resistant Klebsiella pneumoniae

Background: Based on the WHO, multidrug-resistant Klebsiella pneumoniae is a priority pathogen that causes opportunistic infections and is widely spread in the environment. Phage therapy is considered a natural, safe, and very efficient alternative to treat difficult-to-treat infections. Objectives: This study aimed to isolate highly virulent, lytic bacteriophages and evaluate their efficacy for lysing multidrug-resistant K. pneumoniae. Methods: Municipal wastewater samples were collected and filtered using 0.22 µm syringe filters and cultivated with log-phase cultures of K. pneumoniae using enrichment media. After 48 h of incubation, the cultures were centrifuged, and the resultant supernatant was filtered (0.22 µm). The detection of the phage was done using the spot assay with K. pneumoniae as the host. One-step growth kinetics and bacterial reduction tests were conducted to assess the growth kinetics of the isolated phage. The stability of the isolated phage was characterized by subjecting it to various temperature and pH conditions. The chemical stability of the K. pneumoniae phage was determined by exposing it to various organic compounds. A panel of 20 bacterial strains was tested using the spot assay, as well as double agar overlying assay, to determine the host range of the isolated phage. Results: Out of 40 wastewater samples tested, only one sample was tested positive for the K. pneumoniae phage (2.5%) that was lytic against the host strain. The K. pneumoniae phage had a latent period of 15 min and a burst size of 100 virions per infected cell. It was most stable at 37°C and pH range of 6.0 to 10.0. Chemically, the K. pneumoniae phage was resistant to 10% chloroform treatment. Transmission electron micrograph indicated that the K. pneumoniae phage belonged to the order Caudovirales, family Siphoviridae, morphotype B1. Conclusions: Most of the characteristic features of the K. pneumoniae phage indicated the potential of this phage to be used in phage therapy. Hence, a comprehensive study is highly recommended to characterize the K. pneumoniae phage genome, detect its molecular interactions with the host cell, and determine its lytic activity in combination with other phages, which may lead to the efficient utilization of this phage in phage therapy against K. pneumoniae infections.

Methanogenesis suppression and increased power generation in microbial fuel cell during treatment of chloroform containing wastewater

Chloroform is a commonly found solvent in industrial wastewater and is very toxic if present in high concentration. Chloroform degradation in anaerobic processes has been reported earlier and it has also been used as a methanogenesis suppressor in rumen and rice fields. Methanogenesis is undesirable in a microbial fuel cell (MFC) operation, as it results in substrate competition between methanogens and exoelectrogens, thereby reducing the electricity output. Hence, an attempt has been made in this research work to study the effect of chloroform on the performance of MFCs and its effectiveness as methanogenesis suppressor. MFCs with different chloroform dosages in batch as well as continuous mode (0 mM, 8 mM, 16 mM and 22 mM; 0 mM and 22 mM) were operated. High open circuit voltages in batch MFC22 (781 mV) and 53 % higher peak power densities in dosed continuous MFC confirmed its efficiency as a methanogenesis suppressor. Further experimentations showed 74 % lower internal resistance in dosed MFCs. Higher coulombic efficiencies of 31.2 % (MFC22) and 18 % (MFCC22) were noted in dosed reactors than the control reactors (MFC0 – 23.8 % and MFCC0 - 8%). At the end of batches, chloroform concentration was found to be negligible, confirming its degradation in MFC.

Oral Treatment With Ileal Spores Triggers Immunometabolic Shifts in Chicken Gut.

The animal gut is a major site affecting productivity via its role in mediating functions like food conversion and pathogen colonization. Live microorganisms like probiotics are widely used to improve poultry productivity. However, given that chicks receive their microbiota from the environment at-hatch, a bacterial treatment that can stimulate gut immune maturation in early life can benefit animal health. Thus, our lab has begun investigating alternative means to improve poultry health via single inoculation with microbial spores. In this study, we orally-inoculated day-old chicks with ileal scrapings (ISs) enriched for spores via chloroform treatment (SPORE) or non-treated (CON). At 3, 7, and 14 days post-inoculation (dpi), gut permeability was measured via FITC-dextran assay in serum. Additionally, small intestinal scrapings (SISs) were tested for in vitro Salmonella killing and total IgA. Lastly, distal ileum was either fixed or flash-frozen for microscopy or kinome peptide array, respectively. Using bacterial 16S rRNA gene sequencing, SPORE and CON inocula were highly-similar in bacterial composition. However, spores were detected in SPORE but not in CON inoculum. Segmented filamentous bacteria (SFB) filaments were observed in the distal ileum in SPORE birds as early as 3 dpi and all birds at 7 and 14 dpi. Additionally, SFB were detected via PCR in the ceca, colonizing all SPORE birds at 3 dpi. At 3 dpi, SPORE birds exhibited lower gut permeability vs. CON. In SPORE birds, SISs induced greater Salmonella growth in vitro at 3 dpi yet significantly-reduced Salmonella load at 7 and 14 dpi compared to CON in an IgA-independent manner. SPORE distal ileal tissue exhibited unique upregulation of several immunometabolic processes vs. CON birds, including innate (Toll-like receptor, JAK-STAT) and adaptive (T/B cell receptor, TH17 differentiation) immune pathways, PI3K/Akt signaling, mTOR signaling, and insulin-related pathways. Collectively, these data suggest oral inoculation with ileal spores generally-improved gut health.Importance: We report that ileal, spore-forming commensal microbes have potent effects on ileum immunometabolism. Additionally, we identify a functional ileal phenotype in spore-treated chickens, which matched several of the observed immunometabolic changes and was associated with SFB colonization in the ileum.

Characterization and genomic analysis of a ranavirus associated with cultured black-spotted pond frogs (Rana nigromaculata) tadpoles mortalities in China.

Ranaviruses are globally emerging pathogens causing mass mortality in both farmed and wild populations. In this study, we confirmed that a novel ranavirus was related to mass die-offs of black-spotted pond frogs (Rana nigromaculata) in Sichuan Province, China. Histological examination revealed marked degeneration or necrosis in parenchymatous organs, and intracytoplasmic inclusions can be observed in the liver and intestine cells. The virus was isolated from diseased tadpoles (tentatively named the virus as RNRV), and the ranavirus-like particles could be observed in infected EPC cells by the electron microscope. The tadpoles challenged with the isolated virus displayed similar clinical symptoms and pathogenicity as those naturally infected. Physicochemical characteristics showed that it was sensitive to chloroform treatment, trypsin treatment, heating treatment, acidity and alkalinity. Biological characteristics showed that RNRV can induce cytopathic effect (CPE) in various fish cell lines and the optimum growth temperature is 25~28°C. In addition, the complete genome sequence of RNRV was determined (GenBank accession number MG791866) and analysed. The results showed that the genome of RNRV consists of 104,286bp containing 55.2% GC content and 104 predicted ORFs were identified in the genome. Phylogenetic analysis of the complete genome revealed that RNRV should belong to the FV3-like group in genus Ranavirus, and is more closely related to RGV and STIV. These studies confirmed that the RNRV was the causative agent of this natural epizootic event and genome analysis indicated that it belongs to the FV3-like group. In addition, viral physicochemical and biological characteristics will provide a scientific basis for prevention and control.

Effects of polymer residue on the pull-in of suspended graphene

Graphene is an ideal candidate for building microelectromechanical system (MEMS) devices because of its extraordinary electronic and mechanical properties. Some research has been done to study the MEMS pull-in phenomenon in suspended graphene, but no one has yet considered the effects of polymer residue. Polymer residue is an inevitable consequence when transferring polycrystalline graphene (PCG) grown using chemical vapor deposition, the most common graphene growth method. Polymer residue is also introduced when using photolithography to build MEMS devices. In this paper, the authors study the effects of polymer residue on the pull-in of suspended PCG ribbon devices and find that thick polymer residues cause a variation in pull-in voltage. However, after removing most of the polymer residue using a more abrasive chloroform treatment, the authors find that the graphene structure is no longer able to suspend itself as the graphene-substrate interaction energy becomes greater than the strain energy needed to conform graphene to the substrate. Therefore, polymer residue is found to cause variation in the pull-in voltage but is also found to help in graphene’s suspension at high length to displacement ratios.

Effects of olfactory deprivation on action potential and ankyrin-G expression in glutamatergic neurons in the barrel cortex of mice

To investigate effect of upregulated touch sensation induced by olfactory deprivation on action potentials and ankyrin-G expression in the glutamatergic neurons in the barrel cortex of mice. Chloroform (40 μL) was dropped into the unilateral nasal cavity to induce olfactory deprivation in 40 C57 mice (12 days old), whose glutamatergic neurons were genetically labeled with yellow fluorescent protein (YFP). Behavioral experiments were carried out to assess the effects of olfactory deprivation on whisker tactile of the mice. The action potentials of the glutamatergic neurons in the barrel cortex on the side with or without chloroform treatment (olfactory deprivation group and control group, respectively) were recorded by patch-clamp electrophysiological recording, and ankyrin-G expression in the proximal axonal segment of the neurons was detected with immunohistochemistry. Compared with those on the control side, the inter-spike intervals of the barrel glutamatergic neurons were significantly decreased and the absolute refractory periods were significantly shortened on the side with olfactory deprivation (P < 0.01); the expression of ankyrin-G was also significantly increased in the proximal axonal segment of the glutamatergic neurons in the barrel cortex on the side with olfactory deprivation (P < 0.01). Olfaction deprivation induces up-regulation of touch sensation in mice possibly as a result of functional enhancement of the glutamatergic neurons and increased ankyrin-G expression in the barrel cortex.

Structural and magnetic properties of P25 TiO2 nanoparticles doped by Co

The present work reports on the structural and magnetic properties of TiO2 P25 Degussa powder doped with cobalt. Dissolution of cobalt in TiO2 matrix was achieved by precipitation of tris(diethyldithiocarbamato) cobalt(III) (CoDtc - Co[S2CN(C2H5)2]3 in chloroform (CHCl3) and consequent thermal treatment. Five different Co atomic percentage contents were chosen, e.g., 0.5%, 1%, 2%, 5% and 10%. It was shown that cobalt atoms are uniformly distributed across TiO2 matrix and exhibit antiferromagnetic interactions at low temperatures.

Achillea biebersteinii extracts suppress angiogenesis and enhance sensitivity to 5-fluorouracil of human colon cancer cells via the PTEN/AKT/mTOR pathway in vitro

Objective: To investigate the antiproliferative, anti-angiogenic, and apoptotic effects of extracts of Achillea biebersteinii (ABE) and combined treatments of ABE with 5-fluorouracil (5-FU) on HT-29 cells. Methods: The effects of ABE, 5-FU, and combined treatments on the viability of HT-29 cells were determined by 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Isobologram analysis was used to determine synergism between ABE and 5-FU. The apoptotic and anti-angiogenic effects were determined by cell death detection and human vascular endothelial growth factor ELISA method, respectively. Transcriptional and translational expressions of p53, Bax, Bcl-2, p38 MAPK, Akt, PTEN, and mTOR were also evaluated by real-time PCR and Western blotting analysis. Results: ABE decreased the viability of HT-29 cells in a dose-dependent manner. Combined treatment of hexane, chloroform, and methanol extracts of Achillea biebersteinii with 5-FU at IC50 doses decreased the cell viability to 26.0%, 19.1%, and 14.9%, respectively (P Conclusions: Our findings indicate that ABE shows synergism with 5-FU and inhibits the proliferation of HT-29 cells by inducing apoptosis and suppressing angiogenesis, which may provide biological evidence for further use of ABE in the treatment of colorectal cancer.

Effects of different pre-treatment methods on anaerobic mixed microflora for hydrogen production and COD reduction from domestic effluent

Effects of different pretreatment methods on sludge inoculum were evaluated concerning hydrogen (H2) production enhancement and COD (chemical oxygen demand) reduction, using domestic effluent in a batch system. The sludge was taken from a recycled line of the activated sludge reactor. Two types of pretreatment were investigated, heat treatment and chloroform treatment. The experiment was conducted at pH 4-6 and inoculum sizes of microbes were 10%, 20%, and 30% respectively; and experiment without sludge pretreatment was also conducted as control. The result showed that 30% COD reduction was achieved for chloroform pretreatment at pH 3 and 10% inoculum size. For heat treatment, a maximum COD removal of 60% was achieved in the experiment at pH 6 and 10% inoculum size. In chloroform pretreatment, a maximum volume of gas evolved was 3.6 mL, at pH 3 and 20% inoculum size. For heat pretreatment, maximum biogas evolved was 2.1 mL, at pH 3 and 10% inoculum size. The experimental results showed that the pretreatment methods (heat treatment and chloroform treatment) at 35 °C and initial pH 5.5 had a positive influence on H2 production yield and COD removal efficiency during the fermentative H2 production as compared to the control experiments (without pretreatment). Heat treatment method was shown to be a simple and useful method for enhancing both H2 producing and COD removal processes from domestic effluent with highest H2 yield and COD removal efficiency at 0.314 mmol H2/g COD and 86%, respectively.