Molecular evaluation of sheep and goats isolates of Pasteurella multocida and their antibiotic resistance.

Pasteurella multocida exists as a commensal in the upper respiratory tracts of livestock, and poultry, and causes a wide variety of diseases in humans and animals. This study aimed to investigate the incidence of P. multocida by bacteriological and molecular characterization in sheep and goats and screening the existence of capsule-specific genes and their antibiotic resistance pattern. Totally, 1650 nasopharyngeal swabs were collected from apparently healthy sheep and goats and 460 lung tissues were collected from slaughtered animals in Fars province, Iran. All samples were cultured and suspected colonies were examined by biochemical tests, antimicrobial assay and polymerase chain reaction (PCR). Among 165 P. multocida (104 sheep and 61 goats) isolates, the capA, capD, and capB genes were amplified in 98, 48, and 12 isolates, respectively. The occurrence of four virulence-associated genes of P. multocida isolates were determined by PCR. Most isolates harbored the toxA (79.40%) and hgbB genes (70.90%) and 59.40% of isolates had the pfhA gene. Almost half of the isolates (46.10%) contained the tbpA gene. According to the current study, P. multocida capsular type A had the most frequency followed by type D. In addition, the high frequency of tbpA, pfhA, toxA, and hgbB genes revealed that these genes are possibly important in the pathogenesis of P. multocida. Oxytetracycline, enrofloxacin, florfenicol, and tilmicosin were the most effective drugs.

Auto-Segmentation and Classification of Glioma Tumors with the Goals of Treatment Response Assessment Using Deep Learning Based on Magnetic Resonance Imaging.

Glioma is the most common primary intracranial neoplasm in adults. Radiotherapy is a treatment approach in glioma patients, and Magnetic Resonance Imaging (MRI) is a beneficial diagnostic tool in treatment planning. Treatment response assessment in glioma patients is usually based on the Response Assessment in Neuro Oncology (RANO) criteria. The limitation of assessment based on RANO is two-dimensional (2D) manual measurements. Deep learning (DL) has great potential in neuro-oncology to improve the accuracy of response assessment. In the current research, firstly, the BraTS 2018 Challenge dataset included 210 HGG and 75 LGG were applied to train a designed U-Net network for automatic tumor and intra-tumoral segmentation, followed by training of the designed classifier with transfer learning for determining grading HGG and LGG. Then, designed networks were employed for the segmentation and classification of local MRI images of 49 glioma patients pre and post-radiotherapy. The results of tumor segmentation and its intra-tumoral regions were utilized to determine the volume of different regions and treatment response assessment. Treatment response assessment demonstrated that radiotherapy is effective on the whole tumor and enhancing region with p-value ≤ 0.05 with a 95% confidence level, while it did not affect necrosis and peri-tumoral edema regions. This work demonstrated the potential of using deep learning in MRI images to provide a beneficial tool in the automated treatment response assessment so that the patient can obtain the best treatment.

Nanoscale vibration could promote tenogenic differentiation of umbilical cord mesenchymal stem cells.

Regulation of mesenchymal stem cell (MSC) fate for targeted cell therapy applications has been a subject of interest, particularly for tissues such as tendons that possess a marginal regenerative capacity. Control of MSCs' fate into the tendon-specific lineage has mainly been achieved by implementation of chemical growth factors. Mechanical stimuli or 3-dimensional (D) scaffolds have been used as an additional tool for the differentiation of MSCs into tenocytes, but oftentimes, they require a sophisticated bioreactor or a complex scaffold fabrication technique which reduces the feasibility of the proposed method to be used in practice. Here, we used nanovibration to induce the differentiation of MSCs toward the tenogenic fate solely by the use of nanovibration and without the need for growth factors or complex scaffolds. MSCs were cultured on 2D cell culture dishes that were connected to piezo ceramic arrays to apply nanovibration (30-80 nm and 1 kHz frequency) over 7 and 14 d. We observed that nanovibration resulted in significant overexpression of tendon-related markers in both gene expression and protein expression levels, while there was no significant differentiation into adipose and cartilage lineages. These findings could be of assistance in the mechanoregulation of MSCs for stem cell engineering and regenerative medicine applications.

Generation and characterization of two immortalized dermal fibroblast cell lines from the spiny mouse (Acomys).

The spiny mouse (Acomys) is gaining popularity as a research organism due to its phenomenal regenerative capabilities. Acomys recovers from injuries to several organs without fibrosis. For example, Acomys heals full thickness skin injuries with rapid re-epithelialization of the wound and regeneration of hair follicles, sebaceous glands, erector pili muscles, adipocytes, and dermis without scarring. Understanding mechanisms of Acomys regeneration may uncover potential therapeutics for wound healing in humans. However, access to Acomys colonies is limited and primary fibroblasts can only be maintained in culture for a limited time. To address these obstacles, we generated immortalized Acomys dermal fibroblast cell lines using two methods: transfection with the SV40 large T antigen and spontaneous immortalization. The two cell lines (AcoSV40 and AcoSI-1) maintained the morphological and functional characteristics of primary Acomys fibroblasts, including maintenance of key fibroblast markers and ECM deposition. The availability of these cells will lower the barrier to working with Acomys as a model research organism, increasing the pace at which new discoveries to promote regeneration in humans can be made.

Genotypic evaluation of Pasteurella multocida isolated from cattle and sheep by pulsed-field gel electrophoresis.

Pasteurella multocida a Gram-negative bacterium exists as a commensal in the upper respiratory tracts of livestock, and poultry. It is causative agent of a range of diseases in mammals and birds including fowl cholera in poultry, atrophic rhinitis in pigs and bovine hemorrhagic septicemia in cattle and buffalo. This study aimed to isolate P. multocida from sheep and cattle lungs sampled and assessed by bacteriological procedures and pulse field gel electrophoresis (PFGE) characterization. In this study 52 isolated of P. multocida were obtained (2016-2017) from clinically healthy and diseased animals (sheep and cattle) evaluated by PFGE for determining the relationship between them. According to the results of this study 12 sheep isolates had similarities above 94.00% and two cattle isolates showed similarities above 94.00%. When compared between sheep and cattle, most isolates showed a similarity of less than 50.00% indicating the great differences between isolates. It is noteworthy that in the present study, performed by PFGE to determine the type of P. multocida isolates, a very high distinction was made to determine the type of isolates and the relationship between isolates based on fragments in their genome using enzymes.

Comparison of the predictive value of the Helsinki, Rotterdam, and Stockholm CT scores in predicting 6-month outcomes in patients with blunt traumatic brain injuries.

PURPOSE: Despite advances in modern medicine, traumatic brain injuries (TBIs) are still a major medical problem. Early diagnosis of TBI is crucial for clinical decision-making and prognosis. This study aims to compare the predictive value of Helsinki, Rotterdam, and Stockholm CT scores in predicting the 6-month outcomes in blunt TBI patients. METHODS: This cohort study was conducted on blunt TBI patients of 15 years or older. All of them were admitted to the surgical emergency department of Shahid Beheshti Hospital in Kashan, Iran from 2020 to 2021 and had abnormal trauma-related findings on brain CT images. The patients' demographic data such as age, gender, history of comorbid conditions, mechanism of trauma, Glasgow coma scale, CT images, length of hospital stay, and surgical procedures were recorded. The Helsinki, Rotterdam, and Stockholm CT scores were simultaneously determined according to the existing guidelines. The included patients' 6-month outcome was determined using the Glasgow outcome scale extended. M Data were analyzed by SPSS software version 16.0. Sensitivity, specificity, negative/positive predictive value and the area under the receiver operating characteristic curve were calculated for each test. The Kappa agreement coefficient and Kuder Richardson-20 were used to compare the scoring systems. RESULTS: Altogether 171 TBI patients met the inclusion and exclusion criteria, with the mean age of (44.9 ± 20.2) years. Most patients were male (80.7%), had traffic related injuries (83.1%) and mild TBIs (64.3%). Patients with lower Glasgow coma scale had higher Helsinki, Rotterdam, and Stockholm CT scores and lower Glasgow outcome scale extended scores. Among all the scoring systems, the Helsinki and Stockholm scores showed the highest agreement in predicting patients' outcomes (kappa = 0.657, p < 0.001). The Rotterdam scoring system had the highest sensitivity (90.1%) in predicting death of TBI patients, whereas the Helsinki scoring system had the highest sensitivity (89.8%) in predicting the 6-month outcome in TBI patients. CONCLUSION: The Rotterdam scoring system was superior in predicting death in TBI patients, whereas the Helsinki scoring system was more sensitive in predicting the 6-month outcome.

Bioinformatics design of recombinant chimeric protein containing SipD and LptD immunogens and evaluation of its immunogenicity against Salmonella Typhimurium.

The growing emergence of resistant bacteria is the current global concern for the humans and animals. Vaccination could be the desirable method to preventing such infectious diseases. Safe and effective vaccines are urgently needed to manage and prevent Salmonella contamination. Subunit vaccines are safe approaches for the protection against Salmonella spp. The bioinformatics methods were performed to determine the gene structure. Gene cassette (rLPSI) was ordered in pET28a (+), and cloned into E.coli BL21 (DE3), and the recombinant protein was expressed using IPTG (1 mM). Mice were immunized by subcutaneous administration of recombinant protein. Then, the mice were challenged by oral administration of 100LD50 of live S. Typhimurium. The Codon adaptation index of the chimeric gene was multiplied by 0.92. Validation results showed that >90% of residues lie in the desired or extra allowed area of the Ramachandran plot. The recombinant protein (65.9 kDa) was expressed in E.coli. Antibody titers in vaccinated mice were significantly different from those in the control groups. Recombinant protein immunization of the mice provided 90% and 70% protection against 10LD50 and 100LD50 of S. Typhimurium, respectively. In general, the results showed the high efficiency of rLPSI chimeric protein as a protective antigen against S. Typhimurium infection. The rLPSI chimeric protein could be an effective recombinant vaccine candidate against S. Typhimurium infection, but more research is needed.

Effects of chemically EGFR targeting on non-targeted physical cell behaviors in 2D and 3D microfluidic cultures of invasive and non-invasive breast cancer cell lines.

Cancer development comprehends changes in cell structural and physical states. Cancer cells are softer than normal cells, produce higher contractile forces, and migrate more easily. While chemotherapy, targets proteins involved in biological behaviors, it may affect cell physicomechanical state due to the interconnections among signaling pathways. Here we treated non-invasive and invasive breast cancer cell lines by targeting EGRF which modulates major biological behaviors. We quantified migration potential of cancer cells in a microfluidic device, and evaluated expression of proteins associated with physical behaviors. Results indicated significant alterations in physical behaviors, with a higher impact on invasive cells. The anti-cancer synergy between biological and physical behaviors was shown by decreasing actin, vinculin, and myosin II content and altered distribution, limiting cell invasion in 3D collagen structure, accompanied by decreasing cell viability and vimentin expression as the EMT biomarker. The center point of changes in physical behaviors was in cytoskeletal remodeling by chemical treatment, potentially through lower contractile force generation and less development of focal adhesions and stress fibers. The synergy between physical and chemical pathways can be used in enhancing anti-cancer drug efficacy.

Identification of Bordetella bronchiseptica in the throat and nose of dogs and cats by PCR.

B.bronchiseptica is pathogenic for some domestic and wild animals. Due to the importance of this bacterium, its presence in dogs and cats has been investigated using PCR. Pharyngeal and nasal swabs were taken from 135 dogs and 42 cats. Based on the PCR performed on the dogs' samples, in 25/63 (39.68%) pharyngeal samples and 20/59 (33.89%) nasal samples DNA of B. bronchiseptica detected. On the other hand, according to the PCR performed on the cats' samples, in 9/23 (39.13%) pharyngeal samples and 319 (15.78%) nasal samples DNA of B. bronchiseptica was existed. According to the present study, the rate of B. bronchiseptica infection is high among dogs and cats in Iran. Also, due to the fact that the prevalence of this bacterium among pets animals is not exactly known in Iran, necessary measures should be taken for rapid diagnosis and treatment and proper control of the infection.

A novel approach of recombinant laterosporulin production using the N-SH2 domain of SHP-2.

BACKGROUND: The current study was aimed at evaluating the role of the N-SH2 domain of SHP-2 as a partner protein in the expression of a toxic peptide, laterosporulin (LTS). We also investigated its effects on the formation of the disulfide bond and functional folding of the peptide in vitro. The N-SH2-LTS protein was expressed as a His-tagged fusion protein, capable of undergoing enzymatic cleavage. RESULTS: Based on the data presented herein, the total yield of the folded fusion protein from inclusion bodies was found to be about 105 mg/l, demonstrating a high-level of heterologous expression. After enzymatic cleavage, 1.5 mg of the folded recombinant laterosporulin was obtained from each 10 mg of the fusion protein. The purity of the recombinant laterosporulin was analyzed by RP-HPLC, to yield peptides with suitable purity (85%). CONCLUSIONS: Our findings indicated the advantages of using the N-SH2 domain of SHP-2 as a rapid and easy approach not only in producing easy target proteins but also in its function as a chaperone. N-SH2 domain of SHP-2 can influence on the purification of laterosporulin at reasonable yield and in a cost-effective fashion. The N-SH2 domain of SHP-2 as a protein chaperone may be potentially favorable to produce other proteins with disulfide bonds.

Chemical inhibitor anticancer drugs regulate mechanical properties and cytoskeletal structure of non-invasive and invasive breast cancer cell lines: Study of effects of Letrozole, Exemestane, and Everolimus.

Regardless of their target and mechanism, anticancer drugs directly influence biological behavior of cancer cells by activating chemical signaling pathways. Due to the complex interaction between diverse signaling pathways, these drugs may profoundly impact the physical characteristics of cancer cells and regulate their mechanical properties. In this study, the effects of two Aromatase Inhibitor (Letrozole and Exemestane), and one mTOR Inhibitor (Everolimus) on cell mechanical properties, actin content/distribution, and nuclear areas of two invasive and non-invasive breast cancer cell line after 24 h treatment with concentrations previously reported were investigated. While metabolic activity of cell lines was highly affected by drug treatment, significant alterations in Young's modulus of cell bodies, nuclear areas, and actin content and distribution were reported with higher impact on invasive cells. It was concluded that regulation of mechanical behavior of cells by all three drugs emphasizes the cross talk between chemical and physical signaling cascades, and describes a correlation between biological and physical behaviors of cancer cells which might give an insight to a better understanding of mechanisms by which anti-cancer drugs function to enhance their performances.

Molecular characterization of lumpy skin disease virus in Iran (2014-2018).

Lumpy skin disease was first reported in the western provinces of Iran in 2014, and this was followed by several outbreaks throughout the country. In this study, 10 Iranian lumpy skin disease virus (LSDV) samples collected during the period of 2014-2018 were characterized by sequence analysis of the GPCR, LSDV142, and IL10LP genes. Sequence comparison of the respective genes revealed a close relationship between Iranian LSDV isolates and viruses from Asia and Europe. Interestingly, some nucleotide sequence diversity was also observed in the IL10LP genes of the Iranian field isolates.

Altered mechanical properties of actin fibers due to breast cancer invasion: parameter identification based on micropipette aspiration and multiscale tensegrity modeling.

The biophysical properties of cells change with cancer invasion to fulfill their metastatic behavior. Cell softening induced by cancer is highly associated with alterations in cytoskeleton fibers. Changes in the mechanical properties of cytoskeletal fibers have not been quantified due to technical limitations. In this study, we used the micropipette aspiration technique to calculate and compare the viscoelastic properties of non-invasive and invasive breast cancer cells. We evaluated the mechanical properties of actin fibers and microtubules of two cancerous cell lines by using multiscale tensegrity modeling and an optimization method. Cancer invasion caused altered viscoelastic behavior of cells and the results of modeling showed changes in mechanical properties of major cytoskeleton fibers. The stiffness and viscosity constant of actin fibers in non-invasive cells were 1.28 and 2.27 times higher than those of the invasive cells, respectively. However, changes in mechanical properties of microtubules were minor. Immunofluorescent staining of fibers and their quantified distributions confirmed altered actin distribution among two cell lines, in contrast to microtubule distribution. This study highlights the function of cytoskeletal fibers in cancer progression, which could be of interest in designing therapeutic strategies to target cancer progress. Firstly, the viscoelastic behavior of non-invasive and invasive cells is examined with micropipette aspiration tests. A tensegrity model of cells is developed to mimic the viscoelastic behavior of cells, and tensegrity element stiffness is evaluated in an optimization procedure based on micropipette aspiration tests. Finally, by using immunofluorescent staining and confocal imaging, mechanical properties of actin filaments and microtubules of cancer cells are investigated during the course of metastasis.

A phenotypic and molecular investigation of biofilm formation in clinical samples of Pseudomonas aeruginosa.

Pseudomonas aeruginosa is identified as a versatile opportunistic microorganism with metabolic diversity contributing to a wide range of health burdens, especially in immunocompromised patients. This bacterium is the cause of 10 to 20% of nosocomial infections. In this study, we evaluated the phenotypic characterizations of biofilm formation in P. aeruginosa clinical isolates using micro-titer plate assay. Indeed, we estimated the prevalence of QS (rhlI, rhlR, rhlAB, lasB, lasI, lasR, aprA) and virulence genes (pslA and cupA) by PCR. The results showed that among 69% of the isolates forming biofilm, 9% were strong biofilm producers, whereas 13% and 47% of isolates produced moderate and low amounts of biofilm, respectively. All isolates possessed cupA and seven QS genes (rhlI, rhlR, rhlAB, lasB,  lasI, lasR, aprA), while 92% of the isolates possessed the pslA gene. Identification of these genes and their association with biofilm formation can be advantageous in adopting therapeutic methods.

The prevalence rate of sexual violence worldwide: a trend analysis.

BACKGROUND: The purpose of the present study is to showcase the image of Sexual Violence (SV) temporal trends through exploring differences in its prevalence rates during 1990-2017 across 195 countries and territories. METHODS: The SV prevalence rates were derived from the Global Burden of Disease (GBD) database during 1990-2017, worldwide. First, the Latent Growth Model (LGM) was employed for assessing the change in SV prevalence rate over time in Asia, Africa, Europe, North America, South America, Australia & Oceania, for men and women separately. Then, the change in SV prevalence rate over time was determined within countries with high and low Human Development Index (HDI). Finally, the Latent Growth Mixture Models (LGMM) were applied for identifying classes where countries within each class have similar trend of SV prevalence rate over time. RESULTS: The SV prevalence was higher among women than men and decreased in both genders over time across the world. The declining trend in SV prevalence against men is visible in both countries with high and low HDI, but SV prevalence against women in countries with low HDI shows an increase. The findings of LGMM identified six classes of SV prevalence trajectories. LGMM allocated Bermuda into the class with the highest decrease in SV prevalence against men, and Equatorial Guinea and Luxembourg into the class with the highest increase. Other countries had very slow declining trends. In terms of SV prevalence against women, LGMM allocated China, North Korea, and Taiwan into the class with the most increase among the countries in the world. Bermuda, Guyana, Mexico, Nigeria, and Saint Lucia were placed into the class which witnessed the largest decline and Angola, Congo, and Equatorial Guinea were ranked next. The trend in other countries was mostly decreasing. CONCLUSION: Given the high economic and social burden that SV has on victims and societies, the rate of SV in most countries does not seem to have dropped remarkably and requires special attention by relevant policymakers. The SV prevalence rate is highly heterogeneous among world countries which may be due to the definitions and tools used, and more importantly, the culture norms.

Development of a double-recombinant antibody sandwich ELISA for quantitative detection of epsilon toxoid concentration in inactivated Clostridium perfringens vaccines.

BACKGROUND: Epsilon toxin (ETX) causes a commonly fatal enterotoxemia in domestic animals. Also, ETX causes serious economic losses to animal husbandry. In this study, we selected several clones against ETX using repertoires displayed on filamentous phage. Anti-ETX specific clones were enriched by binding to immobilized antigen, followed by elution and re-propagation of phage. After multiple rounds of binding selection, ELISA analysis showed that most isolated clones had high affinity and specificity for ETX. RESULTS: Two recombinant monoclonal antibodies against ETX were isolated by phage display technology. B1 phage VH antibody isolated from DAb library and G2 soluble scFv antibody isolated from Tomlinson I + J libraries have been applied as the capture and detection antibodies for developing an ETX sandwich ELISA test, respectively. CONCLUSIONS: Designed ETX sandwich ELISA could be a valuable tool for quantitative detection of ETX in inactivated commercial vaccines against enterotoxemia.

Leader-following group consensus of discrete-time fractional-order double-integrator multi-agent systems.

The current study concerns with the leader-following group consensus issue for discrete-time fractional-order multi-agent systems. The agents are assumed as fractional-order double-integrators. The interconnection among the agents is characterized by a bidirectional fixed graph. By considering some assumptions on the interaction graph, it is proved that leader-following group consensus will be realized if the controller parameters lie in a specific two-dimensional region. In the beginning, the proposed distributed control structure's ability is evaluated for two groups with two leaders. Then, it is extended to an arbitrary number of groups with an arbitrary number of leaders. Simulation results indicate the efficiency of this distributed control structure.

The effect of Bacillus coagulans on cytotoxicity and apoptosis induced by Salmonella Typhimurium in HT-29 cell culture.

BACKGROUND AND OBJECTIVES: Human epithelial cells have been widely used to study the interaction between intestinal cells and pathogens, in vitro. In this study, the effect of probiotic bacteria Bacillus coagulans and its supernatant on the growth inhibition, cytotoxicity and induction of apoptosis caused by Salmonella Typhimurium and its adhesion to HT-29 cells were investigated. MATERIALS AND METHODS: B. coagulans supernatant was used to obtain the minimum inhibitory concentration. To evaluate the cytotoxicity and percent of apoptotic cells, B. coagulans and its supernatant (2, 4, 6 and 8% concentrations) with S. Typhimurium was added to HT-29 cells. The MTT assay was used in order to evaluate the cytotoxicity. Percent of apoptotic cells was reported using a fluorescence staining method. Additionally, the adhesion of S. Typhimurium to HT-29 cells was investigated. The effect of B. coagulans on the level of adhesion was also studied. RESULTS: The most inhibitory effect was shown at the concentration of 80000 µg/ml supernatant of B. coagulans (54.77% ± 1.43). The simultaneous culture of S. Typhimurium with B. coagulans had the lowest amount of cytotoxicity and induced apoptosis among the all co-culture groups of S. Typhimurium with B. coagulans or its supernatant. The determined cytotoxicity and induced apoptosis were 26.06 % ± 3.79 and 17.63 % ± 2.14 respectively. In the adhesion test, it was observed that B. coagulans can significantly prevent adhesion of S. Typhimurium to HT-29 cell. CONCLUSION: B. coagulans can reduce the adhesion, cytotoxicity and induction of apoptosis caused by S. Typhimurium in HT-29 cells in vitro.

Isolation and identification of Pandoraea spp. From bronchoalveolar lavage of cystic fibrosis patients in Iran.

BACKGROUND: Pandoraea species are gram negative, motile, non-spore forming, rod shaped and oxidase positive, obligate aerobes bacteria, and have one polar flagellum. Most of Pandoraea species are associated with lung infections in cystic fibrosis patients. Cystic fibrosis is the most prevalent autosomal recessive hereditary disease in the world that affects various organs of the body. The main important cause of death in these patients is lung involvement. This study was conducted to isolate and identify Pandoraea bacterium from bronchoalveolar lavage and sputum samples of cystic fibrosis patients in Shiraz, Iran. METHODS: In this research 31 samples of bronchoalveolar lavage and sputum were examined by culture and PCR method. Then confirmed isolates were evaluated for susceptibility to different antibiotics and ability to produce biofilm. RESULTS: The results of this study after cultivation, purification and DNA extraction led to the isolation of 4 Pandoraea bacterium by PCR using specific primers. Antibiotic susceptibility test were indicated all isolates were resistant to gentamicin, amikacin and imipenem and susceptible to ciprofloxacin, trimethoprim-sulfumethoxazole, piperacillin and tetracycline. Ability to create biofilm was indicated by some of Pandoraea isolates. According to findings of this study, ability to synthesis biofilm by Pandoraea isolates and resistance to some antibiotics are very important. CONCLUSIONS: Our study notes the role of P. pnomenusa as an emerging pathogen that can cause chronic lung colonization in CF patients. Identification tools need to be accurate and must be based on molecular techniques. Also our findings should raise awareness about antibiotic resistance in cystic fibrosis patients in Iran and ability of including bacterial agents to produce biofilm is an alarm for public health. Thus clinicians should exercise caution about finding of clinical relevance of this pathogen to the infection and prescribing antibiotics, especially in cases of children infections.

Correlation of the cell mechanical behavior and quantified cytoskeletal parameters in normal and cancerous breast cell lines.

BACKGROUND: Cancerous transformation of cells affects their mechanical behavior and cytoskeleton structure. OBJECTIVE: The objective of this research is to investigate a correlation between mechanical properties and cytoskeletal structure features in cancer cell formation. METHODS: Micropipette aspiration was used to compare mechanical properties of normal (MCF10A) and cancerous (T47D) epithelial breast cell lines. Immunofluorescence and confocal microscopy were employed for staining and imaging F-actin and microtubules, and quantifying their fluorescent intensity, anisotropy and fiber distribution. RESULTS: Results indicated higher F-actin intensity (43%) and anisotropy (50%) in normal cells compared to cancer cells, although there was no difference in the microtubules intensity between cell lines. Furthermore, reductions of cortex thickness and actin layer index (60%) were observed in suspended cancer cells compared to normal cells. Changes in cell physical properties induced by cancer were attributed to microtubules. The arranged fibrous structure of microtubules in normal cells was replaced by a disorganized structure in cancer cells. Cancerous cells were about four times softer with higher creep compliance compared to normal cells. CONCLUSIONS: Results of this study confirmed that alterations in cell mechanical properties induced by cancer are highly correlated with changes in F-actin and microtubule content and arrangement. It is suggested that such changes can enhance our knowledge of cancer initiation and progression.