in vitro Study of Primary Isolation and Culture of Adipose-Derived Stem Cells and Induction of Chondrogenic Differentiation.

Articular cartilage has constrained potential to restore. The mesenchymal stem cellular remedy has presented new treatment possibilities for this circumstance. The experiment aimed to verify the chondrogenic differentiation capacity of rat adipose tissue-derived mesenchymal stem cells (AD-MSCs) in vitro inside the presence or absence of transforming growth factor-beta (TGF-ß1). Rat's subcutaneous adipose tissue minced into a small piece (2-3 mm3) was aseptically collected from the subcutaneous fat under anaesthesia and then digested with collagenase type I (1 mg/mL). Spontaneous chondrogenesis occurred in both AD-MSCs pellet cultures and was similar in both TGF-ß1 treated. The untreated pellet cultures were collected after 21 days. Histological assessment for evaluating the level of proteoglycan by alcin blue staining and immunohistochemistry for detecting the presence of collagen type II. A monoclonal antibody directed against collagen type II. Adipose-derived stem cells (AD-MSCs) isolated from rats were immunophenotyped for the expression of MSCs cell surface markers and was performed by Flow cytometer, which demonstrated AD-MSCs highly expressed CD73 (99.69±2.6%), CD90 (98.11±0.3%), and week expression CD44 (17.15±0.3%). The result of histological staining showed the presence of extracellular matrix (ECM) in the hyaline cartilage. This staining indicated a deposit of "acid mucopolysaccharides" in the proximity of the cells. Additionally, most cells are rounded cells stained positive for the presence of the cells encompassed by extracellular matrix (ECM), which were like chondrocytes as seen from the magnified view, lightly pink stained nuclei, and nuclear fast red stain. However, the immunohistochemistry method demonstrated that the presence of TGF-ß1 decreased the levels of collagen type I and increased the levels of collagen type II. In conclusion, subcutaneous adipose tissue-derived stem cells can be used in cartilage tissue engineering.

Primary Care Providers Underutilize Breast Screening MRI for High-Risk Women.

OBJECTIVE: Supplemental MRI screening for women at high risk for breast cancer is underutilized. Our study assessed how primary care providers in our healthcare network identify high-risk women and recommend high-risk screening breast MRI. METHODS: An electronic survey was distributed to providers in OB/GYN, family, and internal medicine departments between 1/14/19 and 3/22/19. The survey inquired about methods used to assess breast cancer risk, familiarity with the American Cancer Society's definition of high-risk, and whether screening breast MRI is recommended for high-risk women. RESULTS: Response rate was 17% (89/524). After excluding providers who ordered ≤10 mammograms per year, the study included 75 respondents, who mostly ordered 10-1000 mammograms per year and supported annual/biennial screening mammogram starting at age 40-50 years. More providers reported estimating breast cancer risk qualitatively (with family, clinical history, and/or breast density) than quantitatively with risk calculators (73/75, 97% vs 22/75, 29%). A minority of providers (23/75, 31%) correctly defined high lifetime risk. Only 9/75 (12%) providers recommended screening MRI for high-risk women. Use of quantitative risk calculators or ability to correctly define high-risk were not associated with likelihood of recommending MRI screening. More providers had recommended MRI for screening in the setting of dense breasts than for high-risk screening (23/75, 31% vs 9/75, 12%). CONCLUSION: Primary care providers at our institution did not routinely recommend screening MRI for high-risk women. Risk assessment and reporting at the time of mammography may improve MRI utilization and is an opportunity for radiologists to add value and directly participate in patient-centered care.

Impact of Age, Race, and Socioeconomic Status on Women's Perceptions and Preferences Regarding Communication of Estimated Breast Cancer Risk.

RATIONALE AND OBJECTIVES: Performing breast cancer risk assessment at the time of screening mammography has potential to increase high-risk identification, appropriate supplemental screening, and risk management. The study's goal is to investigate women's interest in risk assessment and preferred method of risk communication in a diverse patient population. MATERIALS AND METHODS: Surveys in English and five non-English languages were distributed to women presenting for screening mammography at eight screening mammography facilities between February and May 2019 to assess their interest in risk assessment, preferred method, and level of detail of estimated risk communication in hypothetical scenarios where estimated risks are average and elevated. RESULTS: Among 683 survey respondents, 592 (87%) expressed interest in learning about their estimated lifetime risk of breast cancer. Controlling for age, race/ethnicity, and education, women with higher income were more interested in risk assessment than comparison group (p<0.05). The most preferred method of average risk communication was by a mailed letter accompanying mammographic results (57%), but more women exclusively preferred face-to-face communications of elevated risk than of average risk estimate (191, 28% vs. 128, 19%, p<0.0001). Phone communication was more preferred by younger women, electronic communication was less preferred by older women and those with lower income, and non-Hispanic blacks and older women preferred less detailed communication (p<0.05). CONCLUSION: Sociodemographic factors influence women's interest in risk assessment and preference in risk communication about breast cancer. Screening Mammogram facilities implementing risk assessment should consider risk communication strategies that are most effective for their patient population.

Clean graphene interfaces by selective dry transfer for large area silicon integration.

Here we present a very fast, selective mechanical approach for transferring graphene with low levels of copper contamination from seed wafers on which it was grown to target wafers for very large scale integration (VLSI) electronics. We found that graphene/copper or copper/silicon oxide delamination paths could be selected by slow and faster separation rates, respectively. Thus graphene can be transferred to a target wafer, either exposed or protected by the seed copper layer, which can later be removed by etching. Delamination paths were identified by SEM and Raman spectroscopy. The sheet resistance of the graphene produced by the two approaches was slightly higher than graphene transferred by a PMMA wet-transfer process, indicating reduced impurity doping, and the variation in the sheet resistance values was much lower. Copper contamination levels, quantitatively established by TOF-SIMS, were several orders of magnitude lower than the values for PMMA assisted transfer. In addition, we demonstrated that top-gated transistor devices from our mechanical, delamination transferred graphene exhibited superior transistor behavior to PMMA-assisted wet transfer graphene. The adhesion energy, strength and range of the interactions were quantitatively determined by nonlinear fracture analyses, and suggest that the roughness of the interface between graphene and copper plays an important role with implications for improvements in manufacturing processes.

Three dimensional graphene scaffold for cardiac tissue engineering and in-situ electrical recording.

In this paper, we present a three-dimensional graphene foam made of few layers of CVD grown graphene as a scaffold for growing cardiac cells and recording their electrical activity. Our results show that graphene foam not only provides an excellent extra-cellular matrix (ECM) for the culture of such electrogenic cells but also enables recording of its extracellular electrical activity in-situ. Recording is possible due to graphene's excellent conductivity. In this paper, we present our results on the fabrication of the graphene scaffold and initial studies on the culture of cardiac cell lines such as HL-1 and recording of their real-time electrical activity.

Utilization of graphene electrode in transparent microwell arrays for high throughput cell trapping and lysis.

Here we present a high-throughput, transparent microfluidic device with embedded microwell arrays sandwiched between transparent electrodes made from graphene (at the bottom) and indium tin oxide (at the top) for dielectrophoretic cell trapping and electrical lysis. Graphene suppresses unwanted faradaic reaction effects on the cells and the medium that is typically observed in ITO based electrodes from application of DC field for electrical lysis. This is because graphene is more electrochemically inert than indium tin oxide (ITO) where ITO undergoes reduction-oxidation (redox) reaction in the presence of electrolyte in most standard cell media. This redox process also compromises ITO's electrical properties and optical transparency over multiple use. The presented microfluidic device shows high efficiency for cell trapping and lysis and an electrochemically stable behavior for long operational life.

A new induction period based reaction rate method for determination trace amounts of phenylhydrazine in water samples.

A simple, selective, and sensitive kinetic spectrophotometric method is described for the determination of trace amounts of phenylhydrazine, which is based on its inhibition effect on the reaction between meta cresol purple (MCP) and periodate in the presence of bromide ions. The reaction was monitored spectrophotometrically by measuring the change in absorbance of MCP at 525 nm. The calibration graph was linear in the range of 1.0-10.0 microM. The detection limit (3 sigma) was 0.020 microM. The relative standard deviations for 10 replicate measurements of 3.0, 5.0 and 7.0 microM of phenylhydrazine were 2.0%, 1.4%, and 0.90%, respectively. The potential interfering substances were studied in the presence of phenylhydrazine. The proposed method was applied to the analysis of water samples.

The homogeneity of vanB gene cluster among enterococcal isolates in Iran.

AIMS: This study aimed to analyse the diversity of the vanB gene cluster in enterococcal species isolated from sewage treatment plants (STP) in Tehran, Iran. METHODS AND RESULTS: The enterococcal isolates were collected from three sewage treatment plants in Tehran, Iran, during 2005. A total of 203 enterococcal isolates, collected over six rounds of sampling from three STPs were tested for the presence of vanB gene. Long-PCR showed that amongst the isolates, three Enterococcus faecium, one Enterococcus gallinarum and one Enterococcus casseliflavus harboured the complete vanB gene cluster. Restriction fragment length polymorphism (RFLP) of the vanB1 gene cluster (5900 bp) from the isolates showed an identical pattern to a standard strain of Enterococcus faecalis (V583). None of the isolates were able to transfer the vanB gene in conjugation experiments. Different pulsed-field gel electrophoresis patterns were obtained for the three E. faecium isolates with vanB gene clusters. CONCLUSIONS: Our results indicated that the dissemination of vanB is not widespread in Tehran. Although only a few vanB positive isolates were detected, vanB was found in several enterococcal species. SIGNIFICANCE AND IMPACT OF THE STUDY: In view of the lack of information on vanB resistance genes and their diversity in Iran, knowledge of the global dissemination of vanB genes in Enterococcus spp. is noteworthy.