This document summarizes an in-vitro study that assessed the cytotoxicity of halloysite nanotubes against two cancer cell lines (HepG2 and HCT116) and human peripheral blood lymphocytes. The halloysite nanotubes were first characterized using transmission electron microscopy, which found that over 50% were 500 nm in length, 90% were less than 150 nm in diameter, and over 90% had an aspect ratio of less than 12. In vitro assays then evaluated the cytotoxicity of the halloysite nanotubes against the two cancer cell lines and human lymphocytes at concentrations up to 1000 μg/mL over 24, 48 and 72 hours. The results collectively indicated that the halloysite nanotubes were
Content Cytotoxicity Studies of Colorectal Carcinoma Cells Using Printed Impe...journalBEEI
Monitoring the effectiveness of drugs on cancer cells is crucial for chemotherapeutics studies. In-vitro cell-based biosensors can be used as an alternative for characteristic studies of cells’ response to drugs. Cell-based sensors provide real-time measurements and require smaller sample volumes compared to conventional T-flask measurement methods. This paper presents a biosensor that detects in real-time, impedance variations of human colon cancer, HCT-116 cells when treated with anti-cancer agent, 5-Fluorouracil (5-FU). Two different extracellular matrix (ECM); polyaniline and gelatin were tested and evaluated in terms of attachment quality. Polyaniline was found to provide the best attachment for HCT-116 cells and was used for cytotoxicity studies. Cytokinetic behavior indicated that 5-FU inhibited HCT-116 cells at IC50 of 6.8 µg/mL. Trypan blue exclusion method for testing cell viability was used to validate the impedance measurements, where the cancer cell concentrations were reduced to ~35% when treated with 2.5 µg/mL, and 50% when treated with 6.8 µg/mL. The results generated by the microfabricated impedance biosensor are comparable to the Trypan blue method since both gave similar cell growth trend. It can be concluded that the impedance biosensor has potential to be used as an alternative method in drug testing applications.
1) The study investigated the effects of the tyrosine kinase inhibitors crizotinib and dasatinib on canine osteosarcoma cell lines. It found that dasatinib more effectively suppressed cell viability and inhibited hepatocyte growth factor-induced invasion and migration compared to crizotinib.
2) Exogenous hepatocyte growth factor increased invasion, migration, and matrix metalloproteinase production in the osteosarcoma cell lines.
3) Preliminary results showed that four dogs with osteosarcoma treated with adjuvant dasatinib therapy experienced an apparent clinical benefit.
The document describes research on developing doxorubicin-loaded, folic acid-conjugated multi-walled carbon nanotubes (DOX/FA-MWCNTs) for targeted cancer treatment. The nanotubes were engineered through purification, oxidation, and functionalization processes. Doxorubicin was loaded onto the nanotubes through pi-pi stacking interactions. In vitro studies found the DOX/FA-MWCNTs had high drug loading efficiency, controlled release, and preferential uptake by cancer cells. In vivo studies in tumor-bearing rats showed the DOX/FA-MWCNTs improved pharmacokinetics, biodistribution to tumors, and increased survival time compared to free doxorubicin
The document describes a high-throughput screen that identified small molecule compounds that can enhance the pharmacological effects of oligonucleotides. Several "hit" compounds were discovered that potentiated the actions of splice-switching oligonucleotides, antisense oligonucleotides, and siRNAs in cell culture. The hit compounds preferentially caused the release of fluorescent oligonucleotides from late endosomes. Studies in transgenic mice indicated the hit compounds could enhance the in vivo effects of a splice-switching oligonucleotide without significant toxicity. The findings suggest selected small molecules may help advance oligonucleotide-based therapeutics by modulating intracellular trafficking and endosomal release.
Pluripotent stem cells An in vitro model for nanotoxicityDr. Harish Handral
This document discusses the use of pluripotent stem cells (PSCs) as an in vitro model for assessing nanotoxicity. It notes that existing in vitro and in vivo models have limitations, and that PSCs can differentiate into various cell types and provide a more realistic model that reflects human physiology. PSCs are proposed as a promising alternative platform that could help address current challenges in predicting nanomaterial toxicity and screening new drugs and materials in a reliable and cost-effective way. The review focuses on how induced pluripotent stem cells and embryonic stem cells could be used to establish three-dimensional tissue models for more accurately assessing the hazardous effects of nanomaterials.
This study analyzed changes in gut microbiota diversity and composition between tumor and non-tumor tissues in 23 patients with colorectal cancer. DNA was extracted from paired tumor and non-tumor tissue samples and the V4 region of 16S rRNA genes was sequenced. Alpha and beta diversity analyses were performed to evaluate differences in microbial richness and composition between tissue types and cancer stages. The results provide insights into how the gut microbiota may be involved in colorectal cancer progression.
The molecular characterisation of Escherichia coli K1 isolated from neonatal ...Pauline Ogrodzki
This study characterized 30 E. coli strains isolated from the residual liquid and biofilms of neonatal nasogastric feeding tubes. The E. coli strains clustered into five pulsotypes and sequence types (ST). ST95 was the most common and encoded virulence traits associated with neonatal meningitis. ST95 and other strains were able to attach to and invade intestinal and brain cell lines, and persist in macrophages. The colonization of feeding tubes by these pathogenic E. coli strains poses a potential health risk to neonates.
Development of cell culture samples for drug screening with bone marrow stem ...Apollo Hospitals
Side population (SP) refers to a group of cells, which is capable to efflux Hoechst 33342, a DNA-binding dye. SP cells exist both in normal and tumor tissues. SP abundance is used as an indicator for disease prognosis and drug screening in some studies. Our study concentrates on the factors influencing SP analysis.
Content Cytotoxicity Studies of Colorectal Carcinoma Cells Using Printed Impe...journalBEEI
Monitoring the effectiveness of drugs on cancer cells is crucial for chemotherapeutics studies. In-vitro cell-based biosensors can be used as an alternative for characteristic studies of cells’ response to drugs. Cell-based sensors provide real-time measurements and require smaller sample volumes compared to conventional T-flask measurement methods. This paper presents a biosensor that detects in real-time, impedance variations of human colon cancer, HCT-116 cells when treated with anti-cancer agent, 5-Fluorouracil (5-FU). Two different extracellular matrix (ECM); polyaniline and gelatin were tested and evaluated in terms of attachment quality. Polyaniline was found to provide the best attachment for HCT-116 cells and was used for cytotoxicity studies. Cytokinetic behavior indicated that 5-FU inhibited HCT-116 cells at IC50 of 6.8 µg/mL. Trypan blue exclusion method for testing cell viability was used to validate the impedance measurements, where the cancer cell concentrations were reduced to ~35% when treated with 2.5 µg/mL, and 50% when treated with 6.8 µg/mL. The results generated by the microfabricated impedance biosensor are comparable to the Trypan blue method since both gave similar cell growth trend. It can be concluded that the impedance biosensor has potential to be used as an alternative method in drug testing applications.
1) The study investigated the effects of the tyrosine kinase inhibitors crizotinib and dasatinib on canine osteosarcoma cell lines. It found that dasatinib more effectively suppressed cell viability and inhibited hepatocyte growth factor-induced invasion and migration compared to crizotinib.
2) Exogenous hepatocyte growth factor increased invasion, migration, and matrix metalloproteinase production in the osteosarcoma cell lines.
3) Preliminary results showed that four dogs with osteosarcoma treated with adjuvant dasatinib therapy experienced an apparent clinical benefit.
The document describes research on developing doxorubicin-loaded, folic acid-conjugated multi-walled carbon nanotubes (DOX/FA-MWCNTs) for targeted cancer treatment. The nanotubes were engineered through purification, oxidation, and functionalization processes. Doxorubicin was loaded onto the nanotubes through pi-pi stacking interactions. In vitro studies found the DOX/FA-MWCNTs had high drug loading efficiency, controlled release, and preferential uptake by cancer cells. In vivo studies in tumor-bearing rats showed the DOX/FA-MWCNTs improved pharmacokinetics, biodistribution to tumors, and increased survival time compared to free doxorubicin
The document describes a high-throughput screen that identified small molecule compounds that can enhance the pharmacological effects of oligonucleotides. Several "hit" compounds were discovered that potentiated the actions of splice-switching oligonucleotides, antisense oligonucleotides, and siRNAs in cell culture. The hit compounds preferentially caused the release of fluorescent oligonucleotides from late endosomes. Studies in transgenic mice indicated the hit compounds could enhance the in vivo effects of a splice-switching oligonucleotide without significant toxicity. The findings suggest selected small molecules may help advance oligonucleotide-based therapeutics by modulating intracellular trafficking and endosomal release.
Pluripotent stem cells An in vitro model for nanotoxicityDr. Harish Handral
This document discusses the use of pluripotent stem cells (PSCs) as an in vitro model for assessing nanotoxicity. It notes that existing in vitro and in vivo models have limitations, and that PSCs can differentiate into various cell types and provide a more realistic model that reflects human physiology. PSCs are proposed as a promising alternative platform that could help address current challenges in predicting nanomaterial toxicity and screening new drugs and materials in a reliable and cost-effective way. The review focuses on how induced pluripotent stem cells and embryonic stem cells could be used to establish three-dimensional tissue models for more accurately assessing the hazardous effects of nanomaterials.
This study analyzed changes in gut microbiota diversity and composition between tumor and non-tumor tissues in 23 patients with colorectal cancer. DNA was extracted from paired tumor and non-tumor tissue samples and the V4 region of 16S rRNA genes was sequenced. Alpha and beta diversity analyses were performed to evaluate differences in microbial richness and composition between tissue types and cancer stages. The results provide insights into how the gut microbiota may be involved in colorectal cancer progression.
The molecular characterisation of Escherichia coli K1 isolated from neonatal ...Pauline Ogrodzki
This study characterized 30 E. coli strains isolated from the residual liquid and biofilms of neonatal nasogastric feeding tubes. The E. coli strains clustered into five pulsotypes and sequence types (ST). ST95 was the most common and encoded virulence traits associated with neonatal meningitis. ST95 and other strains were able to attach to and invade intestinal and brain cell lines, and persist in macrophages. The colonization of feeding tubes by these pathogenic E. coli strains poses a potential health risk to neonates.
Development of cell culture samples for drug screening with bone marrow stem ...Apollo Hospitals
Side population (SP) refers to a group of cells, which is capable to efflux Hoechst 33342, a DNA-binding dye. SP cells exist both in normal and tumor tissues. SP abundance is used as an indicator for disease prognosis and drug screening in some studies. Our study concentrates on the factors influencing SP analysis.
This study analyzed changes in the salivary microbiota of 16 patients with oral squamous cell carcinoma (OSCC), 10 with oral leukoplakia (OLK), and 19 healthy controls (HCs). Nonstimulated saliva samples were collected and the bacterial profiles were analyzed using 16S rRNA sequencing. The results showed that patients with OLK and OSCC had differences in the numbers of bacterial operational taxonomical units and diversity compared to HCs. Specifically, the OLK group had higher bacterial richness than HCs. Further, the proportions of certain bacterial genera like Streptococcus, Bacillus, and Haemophilus differed between the patient and control groups. This suggests that changes in the salivary
Scientists screened 18 novel compounds for their ability to kill cancer cells. Several compounds showed potent cytotoxic effects, with IC50 values less than 1 μM on multiple cell lines. Three compounds - TMCOS-3, TMCOS-6, and TMCOS-11 - were found to induce apoptotic cell death through DNA fragmentation and caspase activation. TMCOS-11 was found to specifically inhibit tubulin polymerization and cause cell cycle arrest in the G2/M phase. These findings suggest that some of the compounds may be promising new anti-cancer drugs that work by targeting the microtubule protein tubulin.
Radiotherapy promotes the polarization of tumor-associated macrophages (TAMs) in mice with Lewis lung cancer into anti-tumor M1 macrophages. This is accompanied by increased expression of the long non-coding RNA lincRNA-p21 in the TAMs. TAMs exposed to radiation therapy suppress the viability and invasion of Lewis lung cancer cells in culture. Overexpression of lincRNA-p21 in TAMs enhances their anti-tumor effects, while decreasing lincRNA-p21 reduces the effects of radiation therapy, suggesting lincRNA-p21 plays a key role in the anti-tumor actions of radiotherapy in lung cancer.
This document summarizes a study that synthesized lipid carriers containing the anticancer drug doxorubicin (Dox) and tested their efficacy on HeLa and HCT116 cells. The carriers were around 200 nm in size. Treatment with 1 μM Dox delivered via the new carriers decreased survival of resistant HeLa cells by over 50%, while the blank carriers did not impair survival of sensitive HCT116 cells. This validated assay shows potential for determining efficacy of drug delivery systems.
This document describes a label-free method for cell counting using paramagnetic bead aggregation. When cells are lysed in a chaotropic solution, the released DNA causes paramagnetic beads to aggregate. The extent of aggregation correlates with the amount of DNA and cell number. This allows direct enumeration of cells from crude samples. The method is demonstrated by monitoring bacterial growth and obtaining white blood cell counts from whole blood samples, showing good agreement with standard methods. Specific cell types like CD4+ T cells can also be enumerated using bead-based immunocapture prior to the aggregation step. The method requires only inexpensive equipment and could provide an accessible alternative to more expensive cell counting techniques.
Gene transfer in the liver using recombinant adeno-associated virusJonathan G. Godwin
This document describes methods for delivering gene transfer vectors to the liver using recombinant adeno-associated virus (rAAV). It discusses that intravenous injection of rAAV serotypes results in efficient transduction of the liver. The document provides protocols for preparing rAAV vector samples and delivering the vectors to mouse liver through lateral tail vein injection, retro-orbital sinus injection, or portal vein injection. It notes that tail vein injection is the most common method but retro-orbital sinus injection can be used as an alternative for young or neonatal mice. Precise vector dosing and injection technique are emphasized for achieving effective liver-directed gene transfer.
The document describes a study that isolated and characterized anti-cancer compounds from marine bacteria. Bacteria were isolated from soil samples and their extracts were screened on cancer cell lines. The ethyl acetate extract of one isolate inhibited cancer cell growth by 87.34% and was non-toxic. 16S rRNA sequencing identified the bacterium as Micrococcus luteus. GC-MS analysis identified the active compound as pyrrolo(1,2-alpha)pyrazine-1,4-dione hexahydro-3-(2-methylpropyl). This compound from M. luteus shows potential as a natural anti-cancer agent.
This document discusses several in vitro methods for assessing the cytotoxicity of chemotherapeutic drugs, including assays using brine shrimp, MTT, sulforhodamine B, trypan blue dye, and acridine orange/ethidium bromide staining. It describes maintaining cell lines from cervical carcinoma and breast adenocarcinoma in culture, assessing cell viability, and preserving cells in liquid nitrogen. Specific methods are provided for assays including brine shrimp lethality, MTT, sulforhodamine B, trypan blue dye exclusion, acridine orange/ethidium bromide staining, and DNA fragmentation to evaluate the cytotoxic effects of test compounds on cultured malignant cell lines.
Continuous Exposure to Chrysotile Asbestos Can CauseGhazal Khan
This document summarizes a study that found continuous exposure to chrysotile asbestos can induce transformation of human mesothelial cells through signaling of HMGB1 and TNF-α, similar to effects of crocidolite asbestos exposure. Both asbestos types induced epithelial-to-mesenchymal transition in cells, characterized by downregulation of E-cadherin and phosphorylation/nuclear translocation of β-catenin. While crocidolite exposure induced sustained gene expression changes and HMGB1 release, chrysotile effects returned to baseline within 5-8 weeks. Continuous chrysotile exposure was required to maintain elevated HMGB1 levels, supporting the role of fiber persistence in biological activity.
This research article summarizes a study investigating potential artifacts in 1H NMR-based metabolomic studies on cell cultures due to contaminants from plastic cell culture dishes. The researchers found that brief rinsing or incubation of culture medium in plastic dishes eluted chemicals that could confound assays of certain metabolites. Extraction of "null samples" using perchloric acid, methanol-chloroform, or acetonitrile also produced artifacts from plastic dishes, though to a lesser extent with methanol and acetonitrile. The best practice is to run extraction of blank dishes with every batch of experiments to identify background contamination and provide a reference spectrum.
This document describes a screen of 50,000 compounds to identify inhibitors of hepatocyte growth factor (HGF)-induced epithelial scattering. Several structurally distinct compounds were identified that had not previously been reported to have biological activity. Further analysis revealed that many of the compounds broadly inhibit cancer cell proliferation by arresting cell division in G2/M phase with minimal induction of apoptosis. This effect is consistent with microtubule-targeting agents. Biochemical assays showed that several compounds directly inhibit microtubule polymerization. Some pharmacokinetic properties of the compounds were also assessed.
This research article assessed the cytotoxic activity of essential oils from rosemary, turmeric, and ginger plants against cervical cancer (HeLa) cells. Turmeric (CEO) and ginger (GEO) essential oils exhibited potent cytotoxicity, with CEO having an IC50 of 36.6 μg/mL and GEO an IC50 of 129.9 μg/mL. Cell morphology analysis revealed CEO and GEO treated cells displayed chromatin condensation, membrane blebbing, and cell content leakage at concentrations as low as 32.81 μg/mL for CEO and 32.12 μg/mL for GEO. An Annexin V assay further showed CEO and GEO induced cell death through apoptosis. The results suggest CEO and GEO have cytotoxic effects in vitro
This study investigated the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) and the effectiveness of disinfectants at Khartoum Teaching Hospital in Sudan. Samples were collected from various units and surfaces, and tested for MRSA. The prevalence of MRSA was found to be 25% overall. Some MRSA isolates were also resistant to vancomycin. The effectiveness of four commonly used disinfectants was evaluated against MRSA isolates. Statistical analysis showed two disinfectants were significantly effective against MRSA, while two were not significantly effective. Thus, not all disinfectants used in the hospital were equally effective in controlling MRSA.
In Vitro Cytotoxic Activity of Medicinal Plants Used in the Treatment of CancerIJSTA
This study evaluated the in vitro cytotoxic activity of extracts from six Indian medicinal plants traditionally used to treat cancer. Ethanolic and aqueous extracts of plant parts were tested against four human cancer cell lines. Calotropis procera, Ocimum sanctum and Cannabis sativa extracts showed high cytotoxic activity against the cell lines. The extracts demonstrated selectivity in their activity against different cell types. O. sanctum, C. procera, C. sativa and Trigonella foenum-graecum inhibited over 70% of cell growth, indicating anticancer activity. Chenopodium rubrum was active against colon cell lines. The results provide support for the traditional use of some plants for cancer treatment and identify
This document summarizes a study that evaluated the use of the Line Probe Assay (LPA) for rapid detection of Multi Drug Resistant Tuberculosis (MDR-TB) in Sudan. 300 smear-positive sputum samples were collected from TB patients and tested using LPA, culture-based drug susceptibility testing (DST), and conventional laboratory methods. Results found a high prevalence of MDR-TB in Sudan of 38% by DST and 37.3% by LPA. Comparison of LPA and DST results showed high accuracy of LPA for rapid detection of rifampin and isoniazid resistance with a sensitivity of 98.3% and specificity of 100%. LPA provided results within 2
Atmospheric Exposure to Cr III Powder Causes Genotoxicity in Rattus Norvegicus.inventionjournals
Several chemical elements are responsible for altering the genetic integrity of living beings. The metal Cr stands out in this regard. It exists in two oxidation states, Cr VI and Cr III, and has been investigated as an important environmental and occupational contaminant. Although the former is considered carcinogenic, the latter is classified as safe, even for human use in food supplementation. However, most studies with Cr( III) have been carried out by different routes to how it is occupationally found – in the atmosphere. This study evaluated the genotoxicity of Cr(III) inhaled during 8 hours of exposure to the maximum concentration permitted by ATSDR. Fifteen male Rattus norvegicus were used in this study. There were 3 groups (n=5 per group); these were - group exposed to Cr (III) powder (S), the negative control group (NC) and the positive control group (PC). The animals were exposed to Cr aerosol particles at a flow rate of 9L/min and atmospheric concentration of 500μg/m3 for only 8 hours in this study. An increase in genotoxicity and mutagenicity in the group exposed to the metal powder was observed. These findings suggest that further studies should be carried out in order to establish safe levels of exposure to Cr III in work environments
This document provides information about biocompatibility testing of dental materials. It defines biocompatibility and discusses factors that influence a material's biological response such as location, duration, and stresses. Various in vitro tests are described including cytotoxicity, membrane permeability, and cell function assays using cell cultures. Animal tests involve mucous membrane, skin sensitization, and implantation. Usage tests in animals or humans directly evaluate a material's intended clinical use. The document concludes different testing methods provide information but no single test can predict biocompatibility.
In vitro experiments of prokaryotic and eukaryotic antimicrobial peptide cyto...AI Publications
These proteinaceous molecules, called antimicrobial peptides (AMPs), are a varied collection of antimicrobial peptides. The ability of AMPs to combat gut infections necessitates further study of the AMP-GI tract interaction. These peptides need to be tested in vitro for cytotoxicity before they may be considered for use in clinical infections. Using the MTT conversion assay, neutral red dye absorption assay, and a comparison to vancomycin, researchers examined the cytotoxicity of gallidermin, nisin A, natural magainin peptides, and melittin in two gastrointestinal cell types (HT29 and Caco-2). Sheep erythrocyte hemolytic activity was also studied, and the influence of AMPs on paracellular permeability was assessed using transepithelial resistance (TEER) and TEM. Gallidermin, nisin A, magainin I, magainin II, and melittin were the least cytotoxic AMPs. To our knowledge, only Melittin and NIS caused considerable hemolysis. There are two distinct ways that melittin and nisin differ in their ability to kill bacteria. It was the only AMP that had an effect on the permeability of the paracellular space. Intestinal tight junctions and cell–cell adhesion were destroyed by long-term melittin therapy, as were microvilli, cell debris, and cell–cell adhesion. Antimicrobial activity and low cytotoxicity make Gallidermin a promising therapeutic drug. The antibacterial properties of Melittin are limited, but its ability to transport poorly bioavailable medicines may be useful.
Human pulp cells response to portland cement in vitroNelly Castro
Portland cement showed no cytotoxic effects on cultured human pulp cells, unlike other dental materials tested. Cells exposed to Portland cement attached and proliferated normally. Expression of osteonectin and dentin sialophosphoprotein, markers of mineralization, was induced in cells treated with Portland cement. The results suggest Portland cement is biocompatible and may promote reparative dentin formation making it a potential pulp capping material.
it describes the bony anatomy including the femoral head , acetabulum, labrum . also discusses the capsule , ligaments . muscle that act on the hip joint and the range of motion are outlined. factors affecting hip joint stability and weight transmission through the joint are summarized.
Philippine Edukasyong Pantahanan at Pangkabuhayan (EPP) CurriculumMJDuyan
(𝐓𝐋𝐄 𝟏𝟎𝟎) (𝐋𝐞𝐬𝐬𝐨𝐧 𝟏)-𝐏𝐫𝐞𝐥𝐢𝐦𝐬
𝐃𝐢𝐬𝐜𝐮𝐬𝐬 𝐭𝐡𝐞 𝐄𝐏𝐏 𝐂𝐮𝐫𝐫𝐢𝐜𝐮𝐥𝐮𝐦 𝐢𝐧 𝐭𝐡𝐞 𝐏𝐡𝐢𝐥𝐢𝐩𝐩𝐢𝐧𝐞𝐬:
- Understand the goals and objectives of the Edukasyong Pantahanan at Pangkabuhayan (EPP) curriculum, recognizing its importance in fostering practical life skills and values among students. Students will also be able to identify the key components and subjects covered, such as agriculture, home economics, industrial arts, and information and communication technology.
𝐄𝐱𝐩𝐥𝐚𝐢𝐧 𝐭𝐡𝐞 𝐍𝐚𝐭𝐮𝐫𝐞 𝐚𝐧𝐝 𝐒𝐜𝐨𝐩𝐞 𝐨𝐟 𝐚𝐧 𝐄𝐧𝐭𝐫𝐞𝐩𝐫𝐞𝐧𝐞𝐮𝐫:
-Define entrepreneurship, distinguishing it from general business activities by emphasizing its focus on innovation, risk-taking, and value creation. Students will describe the characteristics and traits of successful entrepreneurs, including their roles and responsibilities, and discuss the broader economic and social impacts of entrepreneurial activities on both local and global scales.
This study analyzed changes in the salivary microbiota of 16 patients with oral squamous cell carcinoma (OSCC), 10 with oral leukoplakia (OLK), and 19 healthy controls (HCs). Nonstimulated saliva samples were collected and the bacterial profiles were analyzed using 16S rRNA sequencing. The results showed that patients with OLK and OSCC had differences in the numbers of bacterial operational taxonomical units and diversity compared to HCs. Specifically, the OLK group had higher bacterial richness than HCs. Further, the proportions of certain bacterial genera like Streptococcus, Bacillus, and Haemophilus differed between the patient and control groups. This suggests that changes in the salivary
Scientists screened 18 novel compounds for their ability to kill cancer cells. Several compounds showed potent cytotoxic effects, with IC50 values less than 1 μM on multiple cell lines. Three compounds - TMCOS-3, TMCOS-6, and TMCOS-11 - were found to induce apoptotic cell death through DNA fragmentation and caspase activation. TMCOS-11 was found to specifically inhibit tubulin polymerization and cause cell cycle arrest in the G2/M phase. These findings suggest that some of the compounds may be promising new anti-cancer drugs that work by targeting the microtubule protein tubulin.
Radiotherapy promotes the polarization of tumor-associated macrophages (TAMs) in mice with Lewis lung cancer into anti-tumor M1 macrophages. This is accompanied by increased expression of the long non-coding RNA lincRNA-p21 in the TAMs. TAMs exposed to radiation therapy suppress the viability and invasion of Lewis lung cancer cells in culture. Overexpression of lincRNA-p21 in TAMs enhances their anti-tumor effects, while decreasing lincRNA-p21 reduces the effects of radiation therapy, suggesting lincRNA-p21 plays a key role in the anti-tumor actions of radiotherapy in lung cancer.
This document summarizes a study that synthesized lipid carriers containing the anticancer drug doxorubicin (Dox) and tested their efficacy on HeLa and HCT116 cells. The carriers were around 200 nm in size. Treatment with 1 μM Dox delivered via the new carriers decreased survival of resistant HeLa cells by over 50%, while the blank carriers did not impair survival of sensitive HCT116 cells. This validated assay shows potential for determining efficacy of drug delivery systems.
This document describes a label-free method for cell counting using paramagnetic bead aggregation. When cells are lysed in a chaotropic solution, the released DNA causes paramagnetic beads to aggregate. The extent of aggregation correlates with the amount of DNA and cell number. This allows direct enumeration of cells from crude samples. The method is demonstrated by monitoring bacterial growth and obtaining white blood cell counts from whole blood samples, showing good agreement with standard methods. Specific cell types like CD4+ T cells can also be enumerated using bead-based immunocapture prior to the aggregation step. The method requires only inexpensive equipment and could provide an accessible alternative to more expensive cell counting techniques.
Gene transfer in the liver using recombinant adeno-associated virusJonathan G. Godwin
This document describes methods for delivering gene transfer vectors to the liver using recombinant adeno-associated virus (rAAV). It discusses that intravenous injection of rAAV serotypes results in efficient transduction of the liver. The document provides protocols for preparing rAAV vector samples and delivering the vectors to mouse liver through lateral tail vein injection, retro-orbital sinus injection, or portal vein injection. It notes that tail vein injection is the most common method but retro-orbital sinus injection can be used as an alternative for young or neonatal mice. Precise vector dosing and injection technique are emphasized for achieving effective liver-directed gene transfer.
The document describes a study that isolated and characterized anti-cancer compounds from marine bacteria. Bacteria were isolated from soil samples and their extracts were screened on cancer cell lines. The ethyl acetate extract of one isolate inhibited cancer cell growth by 87.34% and was non-toxic. 16S rRNA sequencing identified the bacterium as Micrococcus luteus. GC-MS analysis identified the active compound as pyrrolo(1,2-alpha)pyrazine-1,4-dione hexahydro-3-(2-methylpropyl). This compound from M. luteus shows potential as a natural anti-cancer agent.
This document discusses several in vitro methods for assessing the cytotoxicity of chemotherapeutic drugs, including assays using brine shrimp, MTT, sulforhodamine B, trypan blue dye, and acridine orange/ethidium bromide staining. It describes maintaining cell lines from cervical carcinoma and breast adenocarcinoma in culture, assessing cell viability, and preserving cells in liquid nitrogen. Specific methods are provided for assays including brine shrimp lethality, MTT, sulforhodamine B, trypan blue dye exclusion, acridine orange/ethidium bromide staining, and DNA fragmentation to evaluate the cytotoxic effects of test compounds on cultured malignant cell lines.
Continuous Exposure to Chrysotile Asbestos Can CauseGhazal Khan
This document summarizes a study that found continuous exposure to chrysotile asbestos can induce transformation of human mesothelial cells through signaling of HMGB1 and TNF-α, similar to effects of crocidolite asbestos exposure. Both asbestos types induced epithelial-to-mesenchymal transition in cells, characterized by downregulation of E-cadherin and phosphorylation/nuclear translocation of β-catenin. While crocidolite exposure induced sustained gene expression changes and HMGB1 release, chrysotile effects returned to baseline within 5-8 weeks. Continuous chrysotile exposure was required to maintain elevated HMGB1 levels, supporting the role of fiber persistence in biological activity.
This research article summarizes a study investigating potential artifacts in 1H NMR-based metabolomic studies on cell cultures due to contaminants from plastic cell culture dishes. The researchers found that brief rinsing or incubation of culture medium in plastic dishes eluted chemicals that could confound assays of certain metabolites. Extraction of "null samples" using perchloric acid, methanol-chloroform, or acetonitrile also produced artifacts from plastic dishes, though to a lesser extent with methanol and acetonitrile. The best practice is to run extraction of blank dishes with every batch of experiments to identify background contamination and provide a reference spectrum.
This document describes a screen of 50,000 compounds to identify inhibitors of hepatocyte growth factor (HGF)-induced epithelial scattering. Several structurally distinct compounds were identified that had not previously been reported to have biological activity. Further analysis revealed that many of the compounds broadly inhibit cancer cell proliferation by arresting cell division in G2/M phase with minimal induction of apoptosis. This effect is consistent with microtubule-targeting agents. Biochemical assays showed that several compounds directly inhibit microtubule polymerization. Some pharmacokinetic properties of the compounds were also assessed.
This research article assessed the cytotoxic activity of essential oils from rosemary, turmeric, and ginger plants against cervical cancer (HeLa) cells. Turmeric (CEO) and ginger (GEO) essential oils exhibited potent cytotoxicity, with CEO having an IC50 of 36.6 μg/mL and GEO an IC50 of 129.9 μg/mL. Cell morphology analysis revealed CEO and GEO treated cells displayed chromatin condensation, membrane blebbing, and cell content leakage at concentrations as low as 32.81 μg/mL for CEO and 32.12 μg/mL for GEO. An Annexin V assay further showed CEO and GEO induced cell death through apoptosis. The results suggest CEO and GEO have cytotoxic effects in vitro
This study investigated the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) and the effectiveness of disinfectants at Khartoum Teaching Hospital in Sudan. Samples were collected from various units and surfaces, and tested for MRSA. The prevalence of MRSA was found to be 25% overall. Some MRSA isolates were also resistant to vancomycin. The effectiveness of four commonly used disinfectants was evaluated against MRSA isolates. Statistical analysis showed two disinfectants were significantly effective against MRSA, while two were not significantly effective. Thus, not all disinfectants used in the hospital were equally effective in controlling MRSA.
In Vitro Cytotoxic Activity of Medicinal Plants Used in the Treatment of CancerIJSTA
This study evaluated the in vitro cytotoxic activity of extracts from six Indian medicinal plants traditionally used to treat cancer. Ethanolic and aqueous extracts of plant parts were tested against four human cancer cell lines. Calotropis procera, Ocimum sanctum and Cannabis sativa extracts showed high cytotoxic activity against the cell lines. The extracts demonstrated selectivity in their activity against different cell types. O. sanctum, C. procera, C. sativa and Trigonella foenum-graecum inhibited over 70% of cell growth, indicating anticancer activity. Chenopodium rubrum was active against colon cell lines. The results provide support for the traditional use of some plants for cancer treatment and identify
This document summarizes a study that evaluated the use of the Line Probe Assay (LPA) for rapid detection of Multi Drug Resistant Tuberculosis (MDR-TB) in Sudan. 300 smear-positive sputum samples were collected from TB patients and tested using LPA, culture-based drug susceptibility testing (DST), and conventional laboratory methods. Results found a high prevalence of MDR-TB in Sudan of 38% by DST and 37.3% by LPA. Comparison of LPA and DST results showed high accuracy of LPA for rapid detection of rifampin and isoniazid resistance with a sensitivity of 98.3% and specificity of 100%. LPA provided results within 2
Atmospheric Exposure to Cr III Powder Causes Genotoxicity in Rattus Norvegicus.inventionjournals
Several chemical elements are responsible for altering the genetic integrity of living beings. The metal Cr stands out in this regard. It exists in two oxidation states, Cr VI and Cr III, and has been investigated as an important environmental and occupational contaminant. Although the former is considered carcinogenic, the latter is classified as safe, even for human use in food supplementation. However, most studies with Cr( III) have been carried out by different routes to how it is occupationally found – in the atmosphere. This study evaluated the genotoxicity of Cr(III) inhaled during 8 hours of exposure to the maximum concentration permitted by ATSDR. Fifteen male Rattus norvegicus were used in this study. There were 3 groups (n=5 per group); these were - group exposed to Cr (III) powder (S), the negative control group (NC) and the positive control group (PC). The animals were exposed to Cr aerosol particles at a flow rate of 9L/min and atmospheric concentration of 500μg/m3 for only 8 hours in this study. An increase in genotoxicity and mutagenicity in the group exposed to the metal powder was observed. These findings suggest that further studies should be carried out in order to establish safe levels of exposure to Cr III in work environments
This document provides information about biocompatibility testing of dental materials. It defines biocompatibility and discusses factors that influence a material's biological response such as location, duration, and stresses. Various in vitro tests are described including cytotoxicity, membrane permeability, and cell function assays using cell cultures. Animal tests involve mucous membrane, skin sensitization, and implantation. Usage tests in animals or humans directly evaluate a material's intended clinical use. The document concludes different testing methods provide information but no single test can predict biocompatibility.
In vitro experiments of prokaryotic and eukaryotic antimicrobial peptide cyto...AI Publications
These proteinaceous molecules, called antimicrobial peptides (AMPs), are a varied collection of antimicrobial peptides. The ability of AMPs to combat gut infections necessitates further study of the AMP-GI tract interaction. These peptides need to be tested in vitro for cytotoxicity before they may be considered for use in clinical infections. Using the MTT conversion assay, neutral red dye absorption assay, and a comparison to vancomycin, researchers examined the cytotoxicity of gallidermin, nisin A, natural magainin peptides, and melittin in two gastrointestinal cell types (HT29 and Caco-2). Sheep erythrocyte hemolytic activity was also studied, and the influence of AMPs on paracellular permeability was assessed using transepithelial resistance (TEER) and TEM. Gallidermin, nisin A, magainin I, magainin II, and melittin were the least cytotoxic AMPs. To our knowledge, only Melittin and NIS caused considerable hemolysis. There are two distinct ways that melittin and nisin differ in their ability to kill bacteria. It was the only AMP that had an effect on the permeability of the paracellular space. Intestinal tight junctions and cell–cell adhesion were destroyed by long-term melittin therapy, as were microvilli, cell debris, and cell–cell adhesion. Antimicrobial activity and low cytotoxicity make Gallidermin a promising therapeutic drug. The antibacterial properties of Melittin are limited, but its ability to transport poorly bioavailable medicines may be useful.
Human pulp cells response to portland cement in vitroNelly Castro
Portland cement showed no cytotoxic effects on cultured human pulp cells, unlike other dental materials tested. Cells exposed to Portland cement attached and proliferated normally. Expression of osteonectin and dentin sialophosphoprotein, markers of mineralization, was induced in cells treated with Portland cement. The results suggest Portland cement is biocompatible and may promote reparative dentin formation making it a potential pulp capping material.
it describes the bony anatomy including the femoral head , acetabulum, labrum . also discusses the capsule , ligaments . muscle that act on the hip joint and the range of motion are outlined. factors affecting hip joint stability and weight transmission through the joint are summarized.
Philippine Edukasyong Pantahanan at Pangkabuhayan (EPP) CurriculumMJDuyan
(𝐓𝐋𝐄 𝟏𝟎𝟎) (𝐋𝐞𝐬𝐬𝐨𝐧 𝟏)-𝐏𝐫𝐞𝐥𝐢𝐦𝐬
𝐃𝐢𝐬𝐜𝐮𝐬𝐬 𝐭𝐡𝐞 𝐄𝐏𝐏 𝐂𝐮𝐫𝐫𝐢𝐜𝐮𝐥𝐮𝐦 𝐢𝐧 𝐭𝐡𝐞 𝐏𝐡𝐢𝐥𝐢𝐩𝐩𝐢𝐧𝐞𝐬:
- Understand the goals and objectives of the Edukasyong Pantahanan at Pangkabuhayan (EPP) curriculum, recognizing its importance in fostering practical life skills and values among students. Students will also be able to identify the key components and subjects covered, such as agriculture, home economics, industrial arts, and information and communication technology.
𝐄𝐱𝐩𝐥𝐚𝐢𝐧 𝐭𝐡𝐞 𝐍𝐚𝐭𝐮𝐫𝐞 𝐚𝐧𝐝 𝐒𝐜𝐨𝐩𝐞 𝐨𝐟 𝐚𝐧 𝐄𝐧𝐭𝐫𝐞𝐩𝐫𝐞𝐧𝐞𝐮𝐫:
-Define entrepreneurship, distinguishing it from general business activities by emphasizing its focus on innovation, risk-taking, and value creation. Students will describe the characteristics and traits of successful entrepreneurs, including their roles and responsibilities, and discuss the broader economic and social impacts of entrepreneurial activities on both local and global scales.
Strategies for Effective Upskilling is a presentation by Chinwendu Peace in a Your Skill Boost Masterclass organisation by the Excellence Foundation for South Sudan on 08th and 09th June 2024 from 1 PM to 3 PM on each day.
Temple of Asclepius in Thrace. Excavation resultsKrassimira Luka
The temple and the sanctuary around were dedicated to Asklepios Zmidrenus. This name has been known since 1875 when an inscription dedicated to him was discovered in Rome. The inscription is dated in 227 AD and was left by soldiers originating from the city of Philippopolis (modern Plovdiv).
हिंदी वर्णमाला पीपीटी, hindi alphabet PPT presentation, hindi varnamala PPT, Hindi Varnamala pdf, हिंदी स्वर, हिंदी व्यंजन, sikhiye hindi varnmala, dr. mulla adam ali, hindi language and literature, hindi alphabet with drawing, hindi alphabet pdf, hindi varnamala for childrens, hindi language, hindi varnamala practice for kids, https://www.drmullaadamali.com
Chapter wise All Notes of First year Basic Civil Engineering.pptxDenish Jangid
Chapter wise All Notes of First year Basic Civil Engineering
Syllabus
Chapter-1
Introduction to objective, scope and outcome the subject
Chapter 2
Introduction: Scope and Specialization of Civil Engineering, Role of civil Engineer in Society, Impact of infrastructural development on economy of country.
Chapter 3
Surveying: Object Principles & Types of Surveying; Site Plans, Plans & Maps; Scales & Unit of different Measurements.
Linear Measurements: Instruments used. Linear Measurement by Tape, Ranging out Survey Lines and overcoming Obstructions; Measurements on sloping ground; Tape corrections, conventional symbols. Angular Measurements: Instruments used; Introduction to Compass Surveying, Bearings and Longitude & Latitude of a Line, Introduction to total station.
Levelling: Instrument used Object of levelling, Methods of levelling in brief, and Contour maps.
Chapter 4
Buildings: Selection of site for Buildings, Layout of Building Plan, Types of buildings, Plinth area, carpet area, floor space index, Introduction to building byelaws, concept of sun light & ventilation. Components of Buildings & their functions, Basic concept of R.C.C., Introduction to types of foundation
Chapter 5
Transportation: Introduction to Transportation Engineering; Traffic and Road Safety: Types and Characteristics of Various Modes of Transportation; Various Road Traffic Signs, Causes of Accidents and Road Safety Measures.
Chapter 6
Environmental Engineering: Environmental Pollution, Environmental Acts and Regulations, Functional Concepts of Ecology, Basics of Species, Biodiversity, Ecosystem, Hydrological Cycle; Chemical Cycles: Carbon, Nitrogen & Phosphorus; Energy Flow in Ecosystems.
Water Pollution: Water Quality standards, Introduction to Treatment & Disposal of Waste Water. Reuse and Saving of Water, Rain Water Harvesting. Solid Waste Management: Classification of Solid Waste, Collection, Transportation and Disposal of Solid. Recycling of Solid Waste: Energy Recovery, Sanitary Landfill, On-Site Sanitation. Air & Noise Pollution: Primary and Secondary air pollutants, Harmful effects of Air Pollution, Control of Air Pollution. . Noise Pollution Harmful Effects of noise pollution, control of noise pollution, Global warming & Climate Change, Ozone depletion, Greenhouse effect
Text Books:
1. Palancharmy, Basic Civil Engineering, McGraw Hill publishers.
2. Satheesh Gopi, Basic Civil Engineering, Pearson Publishers.
3. Ketki Rangwala Dalal, Essentials of Civil Engineering, Charotar Publishing House.
4. BCP, Surveying volume 1
Main Java[All of the Base Concepts}.docxadhitya5119
This is part 1 of my Java Learning Journey. This Contains Custom methods, classes, constructors, packages, multithreading , try- catch block, finally block and more.
2. F.R. Ahmed et al. / Colloids and Surfaces B: Biointerfaces 135 (2015) 50–55 51
Fig. 1. (a) length distribution of HNTs ranging from <250 nm to >2000 nm. (b) Diameter distribution of HNTs from <50 nm to 250 nm. (c) Aspect ratio (length/diameter) of
HNTs from <3 to >15. Counts of HNTs are given as percentage (%) and standard deviation bars (SD) of total no. (>200) of nanotubes counted per sample (total samples; n = 3).
(d) TEM image of halloysite nanotubes (HNTs).
evaluated in-vitro of their cytotoxic potential at high doses against
two model cell lines (colorectal carcinoma cells HCT116; and hep-
atocellular carcinoma cells HepG2) [21,22] which represent the
earliest entry point and the first accumulating organ, respectively,
for xenobiotics and nanoparticles en-route to systemic circulation
after oral delivery [23,24]. Moreover, cytogenetic toxicity of hal-
loysite nanotubes has also been estimated in this study for the first
time by in-vitro mitotic index assay using peripheral blood human
lymphocytes cultures [25].
2. Materials and methods
2.1. Materials
Halloysite (premium grade; Al2Si2O5(OH)4·2H2O, 99.7%) was
received as a gift from New Zealand China Clays Ltd., (New Zealand)
and further it was sieved (125 m) to separate large agglomerates
[4]. Potassium chloride solution (≥99.5% AT), dimethyl sulfoxide
(DMSO anhydrous ≥99.9) and giemsa stain were purchased from
Sigma–Aldrich Chemical Co., Ltd., (Germany). Methanol (100%,
redistilled) and glacial acetic acid (100%) were procured from
Riedel-de Haen (Germany). Phosphate buffer saline tablets (pH 7.4)
were obtained from MP Biomedicals, LLC (France) and KaryoMAX®
Colcemid Solution (10 g/mL) was purchased from Invitrogen
(USA).
2.2. Culture media
The culture media for HCT116 (colorectal carcinoma cells;
ATCC CCL247) and HepG2 (hepatocellular carcinoma cells; ATCC
HB-8065) cell lines was prepared by supplementing high glu-
cose containing ‘Dulbecco’s modified eagle’s medium (DMEM-high
glucose)’ (GIBCO-Invitrogen, Grand Island, NY) with 10% (v/v)
fetal bovine serum (FBS; GIBCO, USA) and 100 IU/mL of Anti-
AntiR (GIBCO). The culture media for lymphocytes was based
on RPMI-1640 containing l-glutamine (Sigma–Aldrich, Germany)
supplemented with 10% (v/v) fetal bovine serum (GIBCO, USA),
100 IU/mL of Anti-AntiR (GIBCO, USA) and 1.5% phytohaemagglu-
tinin (GIBCO, USA).
2.3. Electron microscopy and particle size distribution
Transmission electron microscopy (TEM) was employed to
determine the length, diameter, and aspect ratio distribution of
halloysite nanotubes (HNTs). The samples were prepared by plac-
ing few drops of aqueous suspension of the halloysite samples
(10 mg/mL) on the carbon coated copper grid and then drying in
the air. TEM was then performed using JEOL JEM-2100 at an accel-
erating voltage of 15 kV. The length, diameter distribution analysis
of halloysite nanotubes was carried out by means of first measur-
ing minimum of 200 nanotubes at different places of grid in each
of the 3 TEM samples prepared using Adobe Acrobat 9 Pro software
3. 52 F.R. Ahmed et al. / Colloids and Surfaces B: Biointerfaces 135 (2015) 50–55
and then sorting and graphing the size distribution using Origin-
Pro 8 software [26,27]. The aspect ratio of HNTs was calculated by
dividing the length of the nanotube by its diameter.
2.4. Statistical analysis
All the graphs in this study were plotted using OriginPro 8
software (OriginLab Corporation, Northampton, MA, USA) while
the statistical calculations were performed with computer soft-
ware IBM SPSS Statistics 22 (SPSS Inc., Chicago, IL, USA). The mean
and standard deviations of treatments in WST-1 were compared
with respective controls by means of student-t test (<0.05) while
one-way ANOVA and Duncan multiple range tests were applied
to calculate the pair-wise variance among different treatments for
mitotic index assay with respect to control (<0.05) [28].
2.5. In-vitro cytotoxicity study
The in-vitro cell viabilities of both, HCT116 (colorectal carcinoma
cells) and HepG2 (hepatocellular carcinoma cells) against halloysite
nanotubes were tested using a WST-1 assay (Biovision, CA, USA)
[29–31]. Initially, cells from continuous passage numbers of 12 and
7, respectively, were each seeded into three 96-well plates (Flat
Bottom Costar, Corning, NY, USA) at a density of 2.5 × 104 cells per
well for HCT116 and 2.0 × 104 cells per well for HepG2. The plates
were then incubated at 37 ◦C, 5% CO2 and 95% humidified air for
24 h. After incubation, the cells were washed with PBS (pH 7.4),
followed by the addition of 100 L of halloysite suspensions in cul-
ture media at various final concentrations of 10, 50, 100, 250, 500,
and 1000 g/mL. Non-treated cells containing only culture media
served as the control. Sample media with halloysite without cells
served as blank. These plates were then further incubated for 24 h,
48 h, and 72 h. The plates especially for 48 h and 72 h incubation
were observed for any exhaustion of culture media periodically
after every 24 h as can be visualized by the change in color and were
replaced gently with fresh media with or without final concentra-
tion of HNTs accordingly in the respective non treated and treated
cells. Afterwards the cells were gently washed thrice with PBS (pH
7.4) and then 100 L of solution of WST-1 dye in culture media
(1:10) was added to each well. The plates were allowed to incu-
bate for an hour in the dark and then the absorbance was taken on
scan mode at a wavelength of 460 nm (620 nm was used as a refer-
ence wavelength) using SpectraMaxM5e (Molecular Devices, USA)
plate reader. For calculations, absorbance values of media contain-
ing wells were subtracted from the values of corresponding treated
wells. The percent value indicating the cell viability was obtained
by dividing values of treated cells by those of untreated cells as
control.
2.6. In-vitro mitotic activity assay
Human blood lymphocytes culture was established after blood
taken (with informed consent) from three healthy, non-smoking
male volunteers who were not exposed to any medicine and radi-
ation in the past 1 month and 6 months, respectively.
The human blood lymphocyte cultures were prepared as
reported by Surrallés et al. [32] and the assay was performed
according to the method mentioned by Eroglu et al. [25] with few
modifications. Initially human venous blood (5 mL) was collected
in an anticoagulant containing vacutainer (5 mL) by venipuncture
[32,33].
Then five tubes containing 0.5 mL of blood were added with
4.5 mL of lymphocyte culture medium at ambient temperature fol-
lowed by incubation for 24 h in an incubation chamber (37 ◦C in 5%
CO2 and 95% humidified air). After that, these tubes were added
with lymphocyte culture medium (control) and four concentra-
tions of halloysite suspensions in culture medium, so that final
concentrations of 10, 100, 500, 1000 g/mL of halloysite may be
achieved. This was followed by incubation at slanting (oblique)
position for further 48 h with occasional gentle shaking of the
tubes. To arrest the cells in metaphase, colcemid solution (100 l;
colchicine 10 g/ml) was added in the sample and incubated for
further 1.5 h. The samples were finally centrifuged at 1000 rpm for
8 min and the resulting pellet was re-suspended with gentle vor-
texing. Pre-warmed (37 ◦C) hypotonic solution (75 mM KCl; 5 mL)
was gradually added into it and incubated in water bath (37 ◦C)
for 20 min followed by centrifugation at 1000 rpm for 8 min. The
supernatant was removed without disturbing the Buffy coat. The
freshly prepared ice cold fixative solution (5 mL; methanol:glacial
acetic acid 3:1) was added gradually followed by centrifugation at
1000 rpm for 8 min. This step was repeated a few times until a clear
pellet was obtained. This pellet was re-suspended in fixative solu-
tion (250 l) and placed overnight at 4 ◦C. The cell suspension was
then dropped (2–3 drops) onto the pre-cleaned cold microscopic
slide. These microscopic slides were air dried, stained with Giemsa
stain (2%, 5 min), washed with de-ionized water and dried at room
temperature. For scoring the cells at least ∼1000 cells/microscopic
slide were counted for the presence of interphase and metaphase
stages in control and various treatments. The images were acquired
at 20× magnification using Nikon compound microscope and pro-
cessed in Windows Photo Gallery.
The mitotic index was calculated according to the following for-
mula,
Mitotic Index % =
Cells in Metaphase
Cells in Metaphase and Interphase
× 100
3. Results and discussion
Halloysite nanotubes were first analyzed for their length, diam-
eter, and aspect ratio distribution by means of TEM. The results
suggest that more than 50% of halloysite tubes were in the length
range of 500 nm and the rest mainly within sub-micrometer size
range (see Fig. 1a) while ∼90% of these had diameter of less than
150 nm with more than 60% below 100 nm (Fig. 1b). Calculation
of the aspect ratios of halloysite nanotubes revealed that almost
all (>90%) had low aspect ratios (<12; Fig. 1c) in contrast to higher
aspects ratios of tens to hundreds and thousands which are known
to exert high toxicity in-vitro [34]. These results are consistent with
other studies reporting the size range of halloysite nanotubes in the
range of 500–2000 nm and aspect ratios from 1 to 10 [1,4].
To evaluate the toxic potential of halloysite nanotubes at higher
concentrations, they were first subjected to WST-1 in-vitro cyto-
toxicity testing against the two cell lines HCT116 and HepG2
representing, respectively, the epithelial lining of the major absorp-
tive site for drugs administered via oral route; and the cells of the
first major organ where the nanomaterials are generally localized
and accumulated after absorption [29,31]. A total of six different
final concentrations of 10, 50, 100, 250, 500, and 1000 g/mL of
halloysite nanotubes were employed for this purpose (see Fig. 2).
Halloysite nanotubes exhibited similar profile against both the cell
lines and statistically relevant decline in cellular viability (100,
250, 500, and 1000 g/mL) was found to be concentration depen-
dent. Interestingly, the usually considered toxic concentrations
of 100, 250 and 500 g/mL for many nanomaterials [35], were
found to have no major anti-proliferative activity upon the two
cell types and only cytotostatic effect was observed at concentra-
tions of 250–500 g/mL, as evidenced by the ∼14–28% inhibition
of proliferation over the course of 72 h incubation in both the
cell lines [36,37]. The highest concentration of halloysite nano-
tubes of 1000 g/mL, however was found to exert significant
anti-proliferative activity in both the cases and significant decline
4. F.R. Ahmed et al. / Colloids and Surfaces B: Biointerfaces 135 (2015) 50–55 53
Fig. 2. WST-1 cytotoxicity assay of HCT116 and HepG2 cells treated with different concentrations of HNTs from 10–1000 g/mL at 24 h, 48 h, and 72 h time periods. (* = p < 0.05
compared to the respective controls; ˛ = p < 0.05 compared to cell viability with preceding concentration; n = 3).
Fig. 3. Graph showing mitotic index (MI) of human peripheral blood lymphocytes treated against various concentrations of HNTs compared to control. (n = 3). Duncan
multiple range test* = p < 0.05. Representative compound microscope image (1000×) of geimsa stained peripheral lymphocytes treated with HNTs (1000 g/mL) and showing
metaphase spread and various cells in interphase (scale bar = 50 m).
in viability (∼48–70% from 24–72 h) was observed which were also
significant as compared to preceding concentrations (see Fig. 3;
˛ = p < 0.05). These results represent similar and comparable cyto-
compatibility and safety profile of halloysite nanotubes with the
results demonstrated by Vergaro et al. against HeLa and MCF-7 cells
[12].
Halloysite nanotubes (HNTs) were further assessed for their
cytogenetic toxicity by determining their activity against human
peripheral lymphocytes by means of mitotic index assay. For this
purpose four different concentrations (10, 100, 500, 1000 g/mL) of
halloysite nanotubes were incubated with the peripheral lympho-
cyte culture. In accordance with the WST-1 assay results against
the two cell lines, mitotic index assay demonstrated slight but
statistically relevant inhibition of proliferation of lymphocytes at
only 1000 g/mL concentration (see Fig. 3 and Table 1). How-
ever, the lower concentrations (10, 100, and 500 g/mL) did not
induce mitotic inhibition. It could be attributed to the fact that
the two cell lines are more sensitive towards nanoparticles as
compared to peripheral lymphocytes, thus even lower concentra-
tions of 250–500 g/mL of nanoparticles exhibit anti-proliferative
effect [38]. This might further substantiate the results of WST-1
assay where only 1000 g/mL concentration was found to exert
cytotoxicity. While discussing the prospects of halloysite nano-
tubes two things must be kept in mind, the first being the fact
that other nanoclay materials are known to exhibit lower toxicity
in-vivo as compared to in-vitro results [14]. Secondly, for vari-
ous conventional dosage forms especially tablets, the diluents are
used in the range of minimum 20% for tablets having large dose
sizes (∼400–500 mg) to maximum 90% in tablets having low dose
sizes (≤25 mg), respectively [39]. If the average percentage quan-
tity of diluent of ∼50% is considered for a conventional large tablet
of 250 mg dose than even this would not account for more than
250 mg (50% of total weight 500 mg) of diluent [40–42]. More-
over, upon dilution in GI fluids this supposedly maximum amount
of 250 mg becomes diluted to a concentration of approximately
125–165 g/mL in an average content of 1.5–2.0 L of gastric secre-
tions and food content especially in postprandial situations [43].
It is also pertinent to discuss that as per the FDA Redbook (2007)
guidance to test the toxicological potential of supposedly nontoxic
ingredients in food, nutrition, and pharmaceuticals, the highest
concentration applicable for insoluble substances which in this
case is 1000 g/mL, was used since at the highest concentration
of 2000 g/mL, recommended for soluble substances, HNTs tend
to sediment significantly and interfere with the testing conditions
[44].
These results are indicative of the potential of halloysite nano-
tubes to be used in various oral drug delivery systems; particularly
as diluent/filler material in tablets, capsules, and suspensions;
without causing toxicity to the absorptive sites and first accumu-
lating organ. This study primarily aims to shed light upon the safety
profile of this novel clay mineral, prevalently used in Chinese tradi-
tional medicine, that could save pharmaceutical industry hundreds
5. 54 F.R. Ahmed et al. / Colloids and Surfaces B: Biointerfaces 135 (2015) 50–55
Table 1
Mitotic index (MI) scoring of human peripheral blood lymphocytes against various concentrations of HNTs compared to control. More than 1000 cells were counted each
time (n = 3).
HNT concentration(g/mL) Human lymphocytes count Metaphase scored Mitotic index(MI) Cumulative (MI)
10 1005 1001 1003 59 61 68 5.35 5.73 5.42 5.50 ± 0.20
100 1000 1000 1010 60 56 58 5.66 5.30 5.43 5.46 ± 0.10
500 1008 1000 1020 60 61 70 5.62 5.75 6.42 5.93 ± 0.25
1000 1020 1026 1000 45 42 42 4.22 3.94 4.03 4.06 ± 0.08*
Control 1000 1000 1000 62 66 57 5.83 6.20 5.39 5.81 ± 0.23
*
Duncan multiple range test (p < 0.05).
of millions of dollars in coming years, especially in generic indus-
try [5]. However, these results must be followed by detailed in-vivo
toxicity evaluation of the nanotubes.
4. Conclusion
Halloysite nanotubes decrease cellular viability of HepG2 and
HCT116 cells in a concentration dependent manner with cyto-
static activity at 250 g/mL and 500 g/mL and cytotoxicity at
1000 g/mL. The mitotic index assay against human peripheral
lymphocytes demonstrates that halloysite nanotubes exert statis-
tically relevant cytogenetic toxicity at only 1000 g/mL by blocking
the passage of cell cycle. The safety profile against the two model
cell lines and human peripheral lymphocytes strongly advocate
and justify for in-vivo toxicity study of halloysite nanotubes (HNTs)
and make an important case for commercial pharmaceutical and
biomedical applications based on earlier evidence in literature of
its nature to sustain the release of loaded drugs.
References
[1] M. Du, B. Guo, D. Jia, Newly emerging applications of halloysite nanotubes: a
review, Polym. Int. 59 (2010) 574–582.
[2] Y.M. Lvov, D.G. Shchukin, H. Möhwald, R.R. Price, Halloysite clay nanotubes
for controlled release of protective agents, ACS Nano 2 (2008) 814–820.
[3] E. Joussein, S. Petit, J. Churchman, B. Theng, D. Righi, B. Delvaux, Halloysite
clay minerals — a review, Clay Miner. 40 (2005) 383–426.
[4] S.R. Levis, P.B. Deasy, Characterisation of halloysite for use as a microtubular
drug delivery system, Int. J. Pharm. 243 (2002) 125–134.
[5] Chi Shi Zhi (Halloysite, Kaolin)—Chinese Herbal Medicine, in: Yin Yang House,
2014.
[6] K. Krejcova, P.B. Deasy, M. Rabiskova, Optimization of diclofenac sodium
profile from halloysite nanotubules, Ceska Slov. Farm. 62 (2013) 71–77.
[7] S.R. Levis, P.B. Deasy, Use of coated microtubular halloysite for the sustained
release of diltiazem hydrochloride and propranolol hydrochloride, Int. J.
Pharm. 253 (2003) 145–157.
[8] J. Forsgren, E. Jamstorp, S. Bredenberg, H. Engqvist, M. Stromme, A ceramic
drug delivery vehicle for oral administration of highly potent opioids, J.
Pharm. Sci. 99 (2010) 219–226.
[9] Y.F. Shi, Z. Tian, Y. Zhang, H.B. Shen, N.Q. Jia, Functionalized halloysite
nanotube-based carrier for intracellular delivery of antisense
oligonucleotides, Nanoscale Res. Lett. 6 (2011) 608.
[10] H. Cornejo-Garrido, A. Nieto-Camacho, V. Gómez-Vidales, M.T. Ramírez-Apan,
P. del Angel, J.A. Montoya, M. Domínguez-López, D. Kibanova, J. Cervini-Silva,
The anti-inflammatory properties of halloysite, Appl. Clay Sci. 57 (2012)
10–16.
[11] Refusal Details as Recorded in OASIS by FDA for Refusal FS2-1092363-7/1/24,
in: Import Refusal Report-US Food and Drug Adminitration, 2009.
[12] V. Vergaro, E. Abdullayev, Y.M. Lvov, A. Zeitoun, R. Cingolani, R. Rinaldi, S.
Leporatti, Cytocompatibility and uptake of halloysite clay nanotubes,
Biomacromolecules 11 (2010) 820–826.
[13] N.G. Veerabadran, R.R. Price, Y.M. Lvov, Clay nanotubes for encapsulation and
sustained release of drugs, Nano 02 (2007) 115–120.
[14] S. Maisanaba, S. Pichardo, M. Puerto, D. Gutiérrez-Praena, A.M. Cameán, A. Jos,
Toxicological evaluation of clay minerals and derived nanocomposites: a
review, Environ. Res. 138 (2015) 233–254.
[15] Y.J. Suh, D.S. Kil, K.S. Chung, E. Abdullayev, Y.M. Lvov, D. Mongayt, Natural
nanocontainer for the controlled delivery of glycerol as a moisturizing agent,
J. Nanosci. Nanotechnol. 11 (2011) 661–665.
[16] N. Verma, E. Moore, W. Blau, Y. Volkov, P. Ramesh Babu, Cytotoxicity
evaluation of nanoclays in human epithelial cell line A549 using high content
screening and real-time impedance analysis, J. Nanopart. Res. 14 (2012)
1–11.
[17] G.I. Fakhrullina, F.S. Akhatova, Y.M. Lvov, R.F. Fakhrullin, Toxicity of halloysite
clay nanotubes in vivo: a Caenorhabditis elegans study, Environ. Sci. Nano 2
(2015) 54–59.
[18] J. Cervini-Silva, A. Nieto-Camacho, E. Palacios, J.A. Montoya, V. Gomez-Vidales,
M.T. Ramirez-Apan, Anti-inflammatory and anti-bacterial activity, and
cytotoxicty of halloysite surfaces, Colloids Surf. B Biointerfaces 111C (2013)
651–655.
[19] M. Guo, A. Wang, F. Muhammad, W. Qi, H. Ren, Y. Guo, G. Zhu, Halloysite
nanotubes, a multifunctional nanovehicle for anticancer drug delivery, Chin. J.
Chem. 30 (2012) 2115–2120.
[20] M. Massaro, C.G. Colletti, R. Noto, S. Riela, P. Poma, S. Guernelli, F. Parisi, S.
Milioto, G. Lazzara, Pharmaceutical properties of supramolecular assembly of
co-loaded cardanol/triazole-halloysite systems, Int. J. Pharm. 478 (2015)
476–485.
[21] J.A. Sergent, V. Paget, S. Chevillard, Toxicity and genotoxicity of nano-SiO2 on
human epithelial intestinal HT-29 line cell, Ann. Occup. Hyg. 56 (2012)
622–630.
[22] K. Kawata, M. Osawa, S. Okabe, In vitro toxicity of silver nanoparticles at
noncytotoxic doses to HepG2 human hepatoma cells, Environ. Sci. Technol. 43
(2009) 6046–6051.
[23] C. Schleh, M. Semmler-Behnke, J. Lipka, A. Wenk, S. Hirn, M. Schaffler, G.
Schmid, U. Simon, W.G. Kreyling, Size and surface charge of gold
nanoparticles determine absorption across intestinal barriers and
accumulation in secondary target organs after oral administration,
Nanotoxicology 6 (2012) 36–46.
[24] B. Ballarin-Gonzalez, F. Dagnaes-Hansen, R.A. Fenton, S. Gao, S. Hein, M. Dong,
J. Kjems, K.A. Howard, Protection and systemic translocation of siRNA
following oral administration of chitosan/siRNA nanoparticles, Mol. Ther.
Nucleic Acids 2 (2013) e76.
[25] H.E. Eroglu, A. Aksoy, E. Hamzaoglu, U. Budak, S. Albayrak, Cytogenetic effects
of nine Helichrysum taxa in human lymphocytes culture, Cytotechnology 59
(2009) 65–72.
[26] W.D. Pyrz, D.J. Buttrey, Particle size determination using TEM: a discussion of
image acquisition and analysis for the novice microscopist, Langmuir 24
(2008) 11350–11360.
[27] John E. Bonevich, Wolfgang K. Haller, Measuring the size of nanoparticles
using transmission electron microscopy (TEM), in: NIST-NCL Joint Assay
Protocol, PCC-7, National Cancer Institute-Frederick, Nanotechnology
Characterization Laboratory, 2010, pp. 2–11.
[28] T. Lialiaris, E. Lyratzopoulos, F. Papachristou, M. Simopoulou, C. Mourelatos, N.
Nikolettos, Supplementation of melatonin protects human lymphocytes
in vitro from the genotoxic activity of melphalan, Mutagenesis 23 (2008)
347–354.
[29] A.R. Kim, F.R. Ahmed, G.Y. Jung, S.-W. Cho, D.-I. Kim, S.H. Um, Hepatocyte
cytotoxicity evaluation with zinc oxide nanoparticles, J. Biomed. Nanotechnol.
9 (2013) 926–929.
[30] S. Dey, M. Das, V.K. Balla, Effect of hydroxyapatite particle size, morphology
and crystallinity on proliferation of colon cancer HCT116 cells, Mater. Sci. Eng.
C 39 (2014) 336–339.
[31] Z. Hanif, F.R. Ahmed, S.W. Shin, Y.-K. Kim, S.H. Um, Size- and dose-dependent
toxicity of cellulose nanocrystals (CNC) on human fibroblasts and colon
adenocarcinoma, Colloids Surf. B: Biointerfaces 119 (2014) 162–165.
[32] J. Surralles, E. Carbonell, R. Marcos, F. Degrassi, A. Antoccia, C. Tanzarella, A
collaborative study on the improvement of the micronucleus test in cultured
human lymphocytes, Mutagenesis 7 (1992) 407–410.
[33] C.E. Alakoc, Halil Erhan, Determining mitotic index in peripheral lymphocytes
of welders exposed to metal arc welding fumes, Turkish J. Biol. 35 (2011)
325–328.
[34] W.S. Journeay, S.S. Suri, H. Fenniri, B. Singh, High-aspect ratio nanoparticles in
nanotoxicology, Integr. Environ. Assess. Manag. 4 (2008) 128–129.
[35] P.C. Ray, H. Yu, P.P. Fu, Toxicity and environmental risks of nanomaterials:
challenges and future needs, J. Environ. Sci. Health C Environ. Carcinog.
Ecotoxicol. Rev. 27 (2009) 1–35.
[36] V.N. Sumantran, Cellular chemosensitivity assays: an overview, Methods Mol.
Biol. (Clifton, N.J.) 731 (2011) 219–236.
[37] A. Narang, D. Desai, Anticancer drug development, in: Y. Lu, R.I. Mahato (Eds.),
Pharmaceutical Perspectives of Cancer Therapeutics, Springer, US, 2009, pp.
49–92.
[38] M.M. Joseph, S.R. Aravind, S. Varghese, S. Mini, T.T. Sreelekha, PST-Gold
nanoparticle as an effective anticancer agent with immunomodulatory
properties, Colloids Surf. B: Biointerfaces 104 (2013) 32–39.
[39] United States Pharmacopeia and National Formulary (USP 32-NF 27), in:
Uniformity of Dosage Units, United States Pharmacopeia Convention,
Rockville, MD, 2011.
6. F.R. Ahmed et al. / Colloids and Surfaces B: Biointerfaces 135 (2015) 50–55 55
[40] Tablets Obtained by Direct Compression, in: G. BASF (Ed.) Generic Drug
Formulations, BASF, 2005.
[41] WHO, Training Workshop on Pharmaceutical Development with focus on
Paediatric Formulations, in: P. Development (Ed.), vol. 2014, World Health
Organization, Tallin, Estonia, 2007, Workshop.
[42] Cellulose, Microcrystalline, in: Raymond C. Rowe, Paul J. Paul Sheskey, Marian
E. Quinn (Eds.), Handbook of Pharmaceutical Excipients, Pharmaceutical Press
and American Pharmaceutical Association, London, 2009, pp. 129–132.
[43] Susan A. Charman, William N. Charman, Oral modified-release delivery
systems, in: Michael J. Rathbone, Jonathan Hadgraft, Michael S. Roberts (Eds.),
Modified-Release Drug Delivery Technology, Marcel Dekker, Inc., New York,
Basel, 2003, pp. 1–10.
[44] Guidance for Industry and Other Stakeholders: Toxicological Principles for the
Safety Assessment of Food Ingredients, REDBOOK 2000, in: U. FDA (Ed.), US
Deparment of Health and Human Services, Silver Spring, MD 2007,
pp. 286.