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1.
Scientifica (Cairo) ; 2024: 4689625, 2024.
Article in English | MEDLINE | ID: mdl-38938544

ABSTRACT

Momordica charantia is well known for its medicinal properties. It has exhibited various pharmacological activities, such as antidiabetic, anti-inflammatory, and antimicrobial activities. Although this plant is used worldwide as a vegetable and medicinal ingredient in herbal medicines, its toxicity studies have not been conducted to date. This study attempts to understand its toxicity. The present study examined the activity of two enzymes, acetylcholinesterase and succinate dehydrogenase, as well as histopathological variations in the liver, intestine, and gills of zebrafish. The results of the acetylcholinesterase assay showed that the concentrations of 40 mg/L and 60 mg/L of the four extracts (leaf and fruit extracts of both varieties) exhibited increased enzyme activity. Interestingly, the leaves of the green fruit variety at a concentration of 60 mg/L showed the highest activity, with a value of 2.824 ± 0.0682 micromoles/min compared to the control value of 1.8347 ± 0.0046 micromoles/min. On the other hand, the succinate dehydrogenase assay revealed that the concentrations of 40 mg/L and 60 mg/L of the extracts decreased the enzyme activity. The highest inhibition was observed in the concentration of 60 mg/L of the leaves of the white-fruited variety and the green-fruited variety, with values of 1.884 ± 0.0482 micromoles/min compared to the control value of 2.747 ± 0.0046 micromoles/min. The studies on histopathological changes also demonstrated abnormalities in the brain, liver, intestine, and gills of zebrafish after the exposure to the extracts of M. charantia. The severity of the damage varied from low to high concentraions. In general, this study sheds light on the safety profile of Momordica charantia and highlights its potential toxicity in animal models. The findings suggest that more research is needed to fully understand the toxicity of this plant and its implications for human use.

2.
MycoKeys ; 104: 9-50, 2024.
Article in English | MEDLINE | ID: mdl-38665970

ABSTRACT

This article presents the results of an ongoing inventory of Ascomycota in Yunnan, China, carried out as part of the research project series "Exploring ascomycete diversity in Yunnan". From over 100 samples collected from diverse host substrates, microfungi have been isolated, identified and are currently being documented. The primary objective of this research is to promote the discovery of novel taxa and explore the ascomycete diversity in the region, utilising a morphology-phylogeny approach. This article represents the second series of species descriptions for the project and introduces three undocumented species found in the families Bambusicolaceae, Dictyosporiaceae and Periconiaceae, belonging to the suborder Massarineae (Pleosporales, Dothideomycetes). These novel taxa exhibit typical morphological characteristics of Bambusicola, Periconia and Trichobotrys, leading to their designation as Bambusicolahongheensis, Periconiakunmingensis and Trichobotryssinensis. Comprehensive multigene phylogenetic analyses were conducted to validate the novelty of these species. The results revealed well-defined clades that are clearly distinct from other related species, providing robust support for their placement within their respective families. Notably, this study unveils the phylogenetic affinity of Trichobotrys within Dictyosporiaceae for the first time. Additionally, the synanamorphism for the genus Trichobotrys is also reported for the first time. Detailed descriptions, illustrations and updated phylogenies of the novel species are provided, and thus presenting a valuable resource for researchers and mycologists interested in the diversity of ascomycetes in Yunnan. By enhancing our understanding of the Ascomycota diversity in this region, this research contributes to the broader field of fungal taxonomy and their phylogenetic understanding.

4.
MycoKeys ; 101: 249-273, 2024.
Article in English | MEDLINE | ID: mdl-38333552

ABSTRACT

Freshwater fungi are highly diverse in China and frequently reported from submerged wood, freshwater insects, herbaceous substrates, sediments, leaves, foams, and living plants. In this study, we investigated two freshwater species that were collected from Yunnan and Guizhou provinces in China. Detailed morphological analysis complemented by multi-gene phylogenetic analyses based on LSU, SSU, ITS, RPB2 and TEF1-α sequences data revealed them to be two new saprobic species, namely Acrogenosporaalangiisp. nov. and Conioscyphayunnanensissp. nov. in their asexual morphs. Additionally, Acrogenosporaalangiisp. nov. is reported for the first time as a freshwater ascomycete associated with the medicinal plant Alangiumchinense (Alangiaceae). Detailed morphological descriptions, illustrations and updated phylogenetic relationships of the new taxa are provided herein.

5.
Indian J Thorac Cardiovasc Surg ; 39(6): 632-635, 2023 Nov.
Article in English | MEDLINE | ID: mdl-37885930

ABSTRACT

We report a case of blow-out-type left ventricular free wall rupture (LVFWR) after acute myocardial infarction, who presented with unstable hemodynamic condition in New York Heart Association (NYHA) functional class IV. Immediately, we performed a successful LVFWR repair with sutureless technique using a glue and expanded polytetrafluoroethylene patch on cardio-pulmonary bypass support. Postoperative period and recovery was uneventful. Over a period of 2-year follow-up, the patient is in NYHA class I and cardiac magnetic resonance imaging showed adequate left ventricular (LV) function and no evidence of LV aneurysm.

6.
ACS Biomater Sci Eng ; 9(11): 6357-6368, 2023 11 13.
Article in English | MEDLINE | ID: mdl-37847169

ABSTRACT

Immortalized liver cell lines and primary hepatocytes are currently used as in vitro models for hepatotoxic drug screening. However, a decline in the viability and functionality of hepatocytes with time is an important limitation of these culture models. Advancements in tissue engineering techniques have allowed us to overcome this challenge by designing suitable scaffolds for maintaining viable and functional primary hepatocytes for a longer period of time in culture. In the current study, we fabricated liver-specific nanofiber scaffolds with polylactic acid (PLA) along with a decellularized liver extracellular matrix (LEM) by the electrospinning technique. The fabricated hybrid PLA-LEM scaffolds were more hydrophilic and had better swelling properties than the PLA scaffolds. The hybrid scaffolds had a pore size of 38 ± 8 µm and supported primary rat hepatocyte cultures for 10 days. Increased viability (2-fold increase in the number of live cells) and functionality (5-fold increase in albumin secretion) were observed in primary hepatocytes cultured on the PLA-LEM scaffolds as compared to those on conventional collagen-coated plates on day 10 of culture. A significant increase in CYP1A2 enzyme activity was observed in hepatocytes cultured on PLA-LEM hybrid scaffolds in comparison to those on collagen upon induction with phenobarbital. Drugs like acetaminophen and rifampicin showed the highest toxicity in hepatocytes cultured on hybrid scaffolds. Also, the lethal dose of these drugs in rodents was accurately predicted as 1.6 g/kg and 594 mg/kg, respectively, from the corresponding IC50 values obtained from drug-treated hepatocytes on hybrid scaffolds. Thus, the fabricated liver-specific electrospun scaffolds maintained primary hepatocyte viability and functionality for an extended period in culture and served as an effective ex vivo drug screening platform to predict an accurate in vivo drug-induced hepatotoxicity.


Subject(s)
Nanofibers , Rats , Animals , Drug Evaluation, Preclinical , Tissue Scaffolds , Hepatocytes/metabolism , Liver , Extracellular Matrix , Collagen/metabolism , Polyesters/pharmacology , Polyesters/metabolism
7.
Front Cell Infect Microbiol ; 13: 1252387, 2023.
Article in English | MEDLINE | ID: mdl-37743866

ABSTRACT

Yunnan, located in southwestern China, is known for its high fungal diversity, and many of which are endemic to the region. As part of our ongoing studies on fungi in Yunnan, we introduce two new genera in Phaeothecoidiellaceae (Mycosphaerellales), to accommodate one Repetophragma-like and another Stomiopeltis-like taxa. Pseudorepetophragma gen. nov. is introduced herein as a monotypic genus to accommodate P. zygopetali comb. nov.(≡ Repetophragma zygopetali), whereas Pseudostomiopeltis gen. nov. is introduced to accommodate Ps. xishuangbannaensis gen. et sp. nov. and Ps. phyllanthi comb. nov.(≡ Stomiopeltis phyllanthi), based on a new collection from Yunnan. In addition, Stomiopeltis sinensis is transferred to Exopassalora as E. sinensis comb. nov. due to its phylogenetic affinity and grouped with E. zambiae, the generic type of Exopassalora. This study provides new insights into the biodiversity of fungal species in this region and adds to our understanding of their ecological roles, as well as the resolution to ambiguous taxa in Phaeothecoidiellaceae.


Subject(s)
Ascomycota , China , Phylogeny , Ascomycota/genetics , Biodiversity
8.
MycoKeys ; 98: 273-297, 2023.
Article in English | MEDLINE | ID: mdl-37539358

ABSTRACT

During a survey of microfungi associated with grasslands and related vegetation types from Yunnan Province in China, various ascomycetous and coelomycetous fungi were isolated. This study reports the discovery of four strains of ascomycetous and coelomycetous fungi from dead stalks of Hypericummonogynum L. (Hypericaceae) and Rubusparvifolius L. (Rosaceae) in the Zhaotong region of Yunnan Province, China. The isolates were characterized using multi-locus phylogenetic analyses and were found to represent a new monophyletic lineage in Melanommataceae (Pleosporales, Dothideomycetes). This new clade was named as Dematiomelanommayunnanense gen. et sp. nov. which consists of both sexual and asexual morphs. The sexual morph is characterized by globose to subglobose ascomata with a central ostiole, cylindrical asci with a pedicel and ocular chamber, and muriform, ellipsoidal to fusiform ascospores. The asexual morph has synanamorphs including both brown, muriform macroconidia and hyaline, round to oblong or ellipsoidal microconidia. These findings contribute to the understanding of fungal diversity in grasslands and related vegetation types in Yunnan Province, China.

9.
Anticancer Agents Med Chem ; 23(17): 1924-1931, 2023.
Article in English | MEDLINE | ID: mdl-37469157

ABSTRACT

BACKGROUND: Indazoles are known for their anti-cancer properties. OBJECTIVE: The current investigation was on the synthesis and evaluation of novel indazole derivatives for their anticancer properties. METHODS: A series of novel indazoles were synthesized and characterized by IR, NMR and LCMS. We performed cytotoxic studies for all synthesized compounds on different cell lines such as HeLa, MCF-7 and EAC using MTT assay. The lead compound was tested further for its anti-tumor and anti-angiogenic effect on EAT tumor model. RESULTS: Amongst the series of compounds synthesized, compound KA8 showed potent antiproliferative effect against Hela, MCF-7 and EAC cell lines with IC50 values 10.4 to 11.5 and 13.5 µM respectively. In addition, our compound KA8 significantly decreased the cell viability, body weight, ascites volume and it also showed superior survival ability of mice compared to control groups. Furthermore, it suppressed the formation of neovasculature in the peritoneum of EAT-bearing mice. CONCLUSION: The findings reveal that the lead compound KA8 possesses potent anti-tumor and anti-angiogenic properties thereby promising it to be developed as a novel anticancer agent with further mechanistic studies.


Subject(s)
Antineoplastic Agents , Carcinoma, Ehrlich Tumor , Animals , Mice , Cell Line, Tumor , Indazoles/chemistry , Ascites/drug therapy , Cell Proliferation , Antineoplastic Agents/chemistry , Carcinoma, Ehrlich Tumor/drug therapy , Drug Screening Assays, Antitumor , Molecular Structure , Structure-Activity Relationship
10.
J Fungi (Basel) ; 9(6)2023 Jun 16.
Article in English | MEDLINE | ID: mdl-37367616

ABSTRACT

Fungi are a large and diverse group of microorganisms, and although the estimated number of species ranges between 2 and 11 million, only around 150,000 species have been described thus far. The investigation of plant-associated fungi is beneficial for estimating global fungal diversity, for ecosystem conservation, and for the continued development of industry and agriculture. Mango, one of the world's five most economically important fruit crops, is grown in over 100 countries and has been demonstrated to have a great economical value. During surveys of mango-associated saprobic fungi in Yunnan (China), we discovered three new species (Acremoniisimulans hongheensis, Chaenothecopsis hongheensis and Hilberina hongheensis) and five new records. The phylogenetic analyses of multi-gene sequences (LSU, SSU, ITS, rpb2, tef1-α and tub2) coupled with morphological examinations were used to identify all the taxa.

11.
J Genet Eng Biotechnol ; 21(1): 9, 2023 Jan 30.
Article in English | MEDLINE | ID: mdl-36715825

ABSTRACT

BACKGROUND: Cisplatin resistance is one of the major contributors to the poor survival rate among head and neck cancer (HNC) patients. Focusing on the protein-protein interaction rather than a single protein could provide a better understanding of drug resistance. Thus, this study aimed to identify hub genes in a complex network of cisplatin resistance associated genes in HNC chemotherapy via a series of bioinformatic tools. METHODS: The genes involved in cisplatin resistance were retrieved from the NCBI gene database using "head and neck cancer" and "cisplatin resistance" as key words. The human genes retrieved were analyzed for their interactions and enriched using the STRING database. The interaction between KEGG pathways and genes was visualized in Cytoscape 3.7.2. Further, the hub gene was identified using the Cytohubba plugin of Cytoscape and validated using UALCAN and Human Protein Atlas database. Validated genes were investigated for the drug-gene interaction using the DGIbd database. RESULTS: Out of 137 genes obtained using key words, 133 were associated with cisplatin resistance in the human species. A total of 150 KEGG pathways, 82 cellular components, 123 molecular functions, and 1752 biological processes were modulated on enrichment analysis. Out of 37 hub genes, CCND1, AXL, CDKN2A, TERT, and EXH2 genes were found to have significant (p < 0.05) mRNA expression and effect on overall survival whereas protein expression was found to be positive for all the significant genes except TERT. Thus, they can be targeted with palbociclib, methotrexate, bortezomib and fluorouracil, sorafenib, dasatinib, carboplatin, paclitaxel, gemcitabine, imatinib, doxorubicin, and vorinostat. CONCLUSION: As the pathogenesis of head and neck cancer is complex, targeting hub genes and associated pathways involved in cisplatin resistance could bring a milestone change in the drug discovery and management of drug resistance which might uplift overall survival among HNC patients.

12.
J Reprod Infertil ; 23(3): 213-223, 2022.
Article in English | MEDLINE | ID: mdl-36415497

ABSTRACT

Background: Chromosomal rearrangements play an important role in infertility. Carriers of chromosomal rearrangements have a lower chance of producing normal or balanced gametes due to abnormal segregation of chromosomes at meiosis, which leads to recurrent spontaneous abortions and infertility. Preimplantation genetic testing for structural chromosome rearrangements (PGT-SR) is offered to couples who have balanced chromosomal rearrangements in order to select embryos with a balanced karyotype prior to implantation, thereby increasing the chances of pregnancy. The purpose of the current study was to assess the outcomes of PGT-SR in patients carrying various balanced chromosomal rearrangements and to assess their clinical pregnancy outcome after in vitro fertilization (IVF). Methods: In this study, infertile couples with balanced chromosomal abnormalities undergoing PGT-SR were retrospectively analyzed at a single fertility center from January 2016 to December 2019. Results: PGT-SR was performed on 87 embryos from 22 couples in whom one partner carried a balanced translocation or an inversion. Fifty-seven (65.5%) of these embryos had unbalanced or sporadic aneuploidies, 30 (34.5%) embryos were normal or chromosomally balanced, which were then transferred in 18 couples. A higher rate of unbalanced translocations in comparison to sporadic aneuploidies was observed in couples with reciprocal translocation. The live birth rate per embryo transfer was found to be 66.6% (12/18). Conclusion: PGT-SR is a useful tool in selecting normal or balanced embryos for transfer in IVF, which could lead to a pregnancy by reducing the chance of miscarriages due to chromosome aneuploidy in couples with balanced chromosomal rearrangements.

13.
Biomimetics (Basel) ; 7(4)2022 Sep 30.
Article in English | MEDLINE | ID: mdl-36278706

ABSTRACT

The major goal of liver tissue engineering is to reproduce the phenotype and functions of liver cells, especially primary hepatocytes ex vivo. Several strategies have been explored in the recent past for culturing the liver cells in the most apt environment using biological scaffolds supporting hepatocyte growth and differentiation. Nanofibrous scaffolds have been widely used in the field of tissue engineering for their increased surface-to-volume ratio and increased porosity, and their close resemblance with the native tissue extracellular matrix (ECM) environment. Electrospinning is one of the most preferred techniques to produce nanofiber scaffolds. In the current review, we have discussed the various technical aspects of electrospinning that have been employed for scaffold development for different types of liver cells. We have highlighted the use of synthetic and natural electrospun polymers along with liver ECM in the fabrication of these scaffolds. We have also described novel strategies that include modifications, such as galactosylation, matrix protein incorporation, etc., in the electrospun scaffolds that have evolved to support the long-term growth and viability of the primary hepatocytes.

14.
J Pharm Bioallied Sci ; 14(Suppl 1): S390-S393, 2022 Jul.
Article in English | MEDLINE | ID: mdl-36110695

ABSTRACT

Aims: We aimed to evaluate and compare the efficacy of moisture-insensitive primer (MIP) Transbond MIP while bonding to wet enamel surface. Materials and Methods: This study was done on 100 freshly extracted human premolar teeth, collected from the Department of Oral Surgery, Government Dental College, Calicut. They were stored in distilled water at room temperature. All teeth were healthy and without caries lesion, no evidence of surface defects, or any developmental morphological aberrations. Teeth were divided into 5 groups of twenty each for the testing and Brackets were bonded onto the tooth surface in 5 different bonding sequences using Transbond XT and Transbond MIP.. Results: All the three groups of MIP have gained statistically significant higher SBS compared to Group I (Transbond XT primer in wet). Marked decrease in SBS of Transbond XT resin in wet environment was deduced. MIP in repriming sequence (Group IV) is found to have higher bond strength than other MIP groups. Conclusions: Clinical acceptability of Transbond MIP in wet environment could be recommended. MIP repriming should be done in cases where there is a doubt of contamination with saliva after initial primer application.

15.
ACS Omega ; 7(35): 31377-31387, 2022 Sep 06.
Article in English | MEDLINE | ID: mdl-36092608

ABSTRACT

Cereals and pulses comprise the largest proportion in a typical Indian diet plate. This research mainly focuses on determining the nutritional composition, bioactive compound characterization, and antioxidant activities of seven selected cereals and pulses. The total carbohydrate content was high in unripe banana (67.65/100 g) and arrowroot (63.76/100 g). Finger millet (44.55 µmol %), chickpea (53.33 µmol %), and green gram (17.40 µmol %) showed high oleic, linoleic, and linolenic acid contents, respectively. The ascorbic acid content was the highest in chickpea and horse gram at 86.83 and 83.76 mg/100 g, respectively. The major phenolics and flavonoids quantified and confirmed using HPLC and UHPLC-HRMS/MS were gallic, protocatechuic, vanillic, para-coumaric, ferulic, chlorogenic, sinapic, and trans-cinnamic acids, rutin, and quercetin. The sample extracts showed dose-dependent antioxidant activity to combat the reactive oxygen species. Hence, these serve as an excellent source for the development of functional food formulations for lowering the risk of various diseases.

16.
ACS Omega ; 7(36): 31713-31725, 2022 Sep 13.
Article in English | MEDLINE | ID: mdl-36120066

ABSTRACT

Printed circuit boards (PCBs) constitute an important segment of electronic waste that can be effectively utilized to recover valuable metals and organics. The present work is focused on the kinetics and product distribution from pyrolysis of three different PCB samples, viz., television PCB (TV PCB), motherboard PCB (MB PCB), and hard disk PCB (HD PCB). The PCBs were pretreated to eliminate most of the metallic constituents. Kinetic analysis was performed using Vyazovkin's isoconversional method and distributed activation energy model (DAEM). The average apparent activation energies obtained from the Vyazovkin method were 207.2, 158.9, and 179.7 kJ mol-1 for the TV PCB, MB PCB, and HD PCB, respectively. The DAEM with five, four, and four pseudo-components was used to describe the decomposition kinetics of the TV PCB, MB PCB, and HD PCB, respectively. Importantly, two types of distributions, viz., Gaussian and Weibull, were utilized to effectively model the nonisothermal data obtained from thermogravimetric analysis at 10 and 20 °C min-1. The evolution of pyrolysates belonging to functional groups such as phenolics, aromatics, aliphatics, halogenated compounds, N-containing compounds, and oxygenates was studied at two different temperatures (500 and 700 °C) using analytical pyrolysis-gas chromatograph/mass spectrometry (Py-GC/MS). The Py-GC/MS results demonstrated an increase in selectivity to aromatics and straight-chain aliphatics at 700 °C with a concomitant decrease in selectivity to phenols and oxygenates.

17.
J Fungi (Basel) ; 8(9)2022 Aug 24.
Article in English | MEDLINE | ID: mdl-36135624

ABSTRACT

The genus Alternaria is comprised of well-known plant pathogens causing various important diseases in plants, as well as being common allergens in animals and humans. Species of Alternaria can be found as saprobes associated with various dead plant materials. This research aims to enhance the taxonomy of saprobic species in the genus Alternaria found on grasses and herbaceous plants from Italy, based on multi-locus phylogenetic analyses of a concatenated ITS, LSU, SSU, tef1-α, rpb2, gapdh and Alt-a1 DNA sequence dataset combined with morphological characteristics. Multi-locus phylogenetic analyses demonstrated six novel species belonging to the genus Alternaria sect. Alternaria as: A. muriformispora sp. nov., A. obpyriconidia sp. nov., A. ovoidea sp. nov., A. pseudoinfectoria sp. nov., A. rostroconidia sp. nov. and A. torilis sp. nov. Detailed morphological descriptions, illustrations and an updated phylogenetic relationship of taxa in the genus Alternaria sect. Alternaria are provided herein.

18.
Indian J Otolaryngol Head Neck Surg ; 74(Suppl 1): 164-169, 2022 Aug.
Article in English | MEDLINE | ID: mdl-36032922

ABSTRACT

Cisplatin is a chemotherapeutic agent effective against head and neck carcinoma but unfortunately it is cochleotoxic. This study has been designed to investigate the efficacy of OAE in identifying early effects of cisplatin on the cochlea and the importance of protocol for audiological monitoring of cisplatin induced ototoxicity. This is a prospective observational study conducted from October 2012 to September 2014 on 70 patients, receiving Cisplatin for various malignant conditions. Audiological criteria for ototoxicity was considered as a difference of 10 d B or more in pure tone thresholds of two or more adjacent frequencies in conventional audiometry and in DPOAE-Signal noise ratio less than 6 dB or DPOAE amplitude less than 20 dBSPL (irrespective of SNR > 6 dB). According to PTA, 60.7% patients showed ototoxicity after completion of chemotherapy. In DPOAE, according to SNR and amplitude criteria more than 60% patients showed ototoxicity after first cycle of cisplatin at high frequencies (4-8 kHz). DPOAEs is a sensitive tool for early detection of ototoxicity and protocol is necessary for monitoring ototoxicity in patients receiving cisplatin to improve the quality of life.

19.
J Fungi (Basel) ; 8(7)2022 Jun 26.
Article in English | MEDLINE | ID: mdl-35887428

ABSTRACT

Arthrobotrys is the most complex genus of Orbiliaceae nematode-trapping fungi. Its members are widely distributed in various habitats worldwide due to their unique nematode-trapping survival strategies. During a survey of nematophagous fungi in Yunnan Province, China, twelve taxa were isolated from terrestrial soil and freshwater sediment habitats and were identified as six new species in Arthrobotrys based on evidence from morphological and multigene (ITS, TEF, and RPB2) phylogenetic analyses. These new species i.e., Arthrobotrys eryuanensis, A. jinpingensis, A. lanpingensis, A. luquanensis, A. shuifuensis, and A. zhaoyangensis are named in recognition of their places of origin. Morphological descriptions, illustrations, taxonomic notes, and a multilocus phylogenetic analysis are provided for all new taxa. In addition, a key to known species in Arthrobotrys is provided, and the inadequacies in the taxonomic study of nematode-trapping fungi are also discussed.

20.
Micromachines (Basel) ; 13(6)2022 Jun 19.
Article in English | MEDLINE | ID: mdl-35744583

ABSTRACT

Particle synthesis has seen significant advances in current trends. However, the synthesis of metal particles without oxidation is a challenge for researchers. The current study presents a straightforward, convenient, and convincing approach for manufacturing copper (Cu) particles free of surface oxide. The die-sink Electrical Discharge Machine (EDM) of copper alloys with oleic acid resulted in the formation of Cu particles with diameters between 10 to 20 µm. X-ray diffraction (XRD) was used for particle examination after cleaning and sonication with distilled water. Cu particles with oleic acid coating retained a Cu phase without oxidation after synthesis. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were used to determine the size and morphology of generated particles. Fourier transforms infrared (FT-IR) analysis revealed the oleic acid-coated Cu surface bonded with an oxygen atom. Also, the agglomeration and change of size involving Cu particles with increasing voltages in the pulse supply in EDM were reported.

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