34 research outputs found
Real-Time Measurement of the Mitochondrial Bioenergetic Profile of Neutrophils
No full textReal-Time Measurement of the Mitochondrial Bioenergetic Profile of Neutrophils
Citation:
Pulikkot, S., Zhao, M., Fan, Z. Real-Time Measurement of the Mitochondrial Bioenergetic Profile of Neutrophils. J. Vis. Exp. (196), e64971, doi:10.3791/64971 (2023)
ESTABLISHMENT OF A 3D CULTURE MODEL OF GASTRIC STEM CELLS SUPPORTING THEIR DIFFERENTIATION INTO MUCOUS CELLS USING MICROFIBROUS POLYCAPROLACTONE SCAFFOLD
No full textIn the stomach, epithelial stem cells are responsible for glandular homeostasis and continuous production of four main cell lineages secreting mucus, acid, pepsinogen and hormones. While alteration in the proliferation and differentiation program of these stem cells is linked to the origin of gastric cancer, they represent an effective target for chemotherapy and a source for cell therapy or tissue engineering in cases of gastric mucosal damage or loss. The aims of this study were 1) to manufacture various forms of scaffolds using a biodegradable polymer (polycaprolactone), 2) to test the suitability of these scaffolds for growth of mouse gastric stem (mGS) cells, and 3) to evaluate whether this culture system could sustain exposure to acidic environment for possible future applications. Three forms of polycaprolactone scaffold were fabricated: nonporous, microporous and microfibrous. Scanning electron microscopy (SEM) and mechanical testing revealed some similarities between the microfibrous scaffold and extracellular matrix of mouse stomach wall. Examination of mGS cells seeded on different forms of scaffold for 3 days using SEM and calcein viability assay revealed their preferential growth on microfibrous scaffolds fabricated by electrospinning technique. Analysis of the growth pattern of mGS cells on microfibrous scaffolds following 3, 6, 9 and 12 days of culture using SEM and DNA PicoGreen assay demonstrated an initial increase in cell number, followed by reduction by days 9 and 12. To test whether this reduction was associated with cell differentiation, cryosections of cultured mGS cells on scaffolds were probed with gastric epithelial cell differentiation markers. On day 3, none of the markers bound to the cells. However by day 9, approximately, 50% of the cells bound to N-acetyl-Dvii glucosamine-specific lectin (Griffonia simplicifolia II) suggesting differentiation into gland mucous cells. This finding was confirmed by the expression of trefoil factor 2 using immunocytochemisty. In addition, gene expression analysis using quantitative reverse transcription polymerase chain reaction (qRT-PCR) demonstrated that the expression of transcription factor SPDEF, required for differentiation of mucous cells, was gradually up-regulated with culture of mGS cells from 3 to 12 days. To test whether this 3D culture system could tolerate the acidic environment of the stomach, the mechanical/chemical integrity of microfibrous scaffolds and cultured mGS cells were studied at acidic pH (3.0 to 7.4) using tensile strength measurements, fourier transform infrared spectroscopy, calcein assay, and mRNA/protein expression analysis. The in vitro wound-healing assay was also used to examine effects of acidic pH on cell migration. RPMI culture media at pH 3.0 and 4.5 reduced the mechanical integrity of scaffolds and significantly inhibited cell viability by \u3e70%. However, at pH 5.5 and 6.0, no significant change in cell viability and scaffold integrity was observed, but cell migration was inhibited by more than 50%. Interestingly, only after 3-day culture at pH 5.5, N-acetyl-Dglucosamine- specific lectin binding combined with significant up-regulation in the expression of SPDEF gene confirmed mucous cell differentiation. In conclusion, a 3D culture model of mGS cells using microfibrous PCL scaffold supporting their differentiation into gland mucous cells has been established. Reducing the pH value of culture media to 5.5 modulates proliferation/migration programs of mGS cells and speeds up their differentiation into mucous cells. This study provides important basic information for the possible use of mGS cells and microfibrous PCL scaffolds for future gastric tissue engineering studies and viii regenerative therapy of some stomach diseases involving gastric mucosal damage or loss
Sunitha Krishnan i jej walka z seksualnym niewolnictwem kobiet w Indiach
No full textThis article introduces the silhouette of Hindu activist Sunitha Krishan, has been strongly committed to the fight against sexual slavery in India for 20 years. It presents the circumstances of the fact that she started her business, initiatives which was undertaken by her and what was the results of her activities. In summary form the article presents stories that Krishnan has collected by seeing with victims – women and children. The author also essays the presentation of Hindu models of femininity and she demonstrates their influence on the current situation of women. This text is largery based on online sources, articles published on the information portals, interviews with Krishnan and record during the conference with her participation.Niniejszy artykuł przybliża sylwetkę hinduskiej aktywistki Sunithy Krishan, która od 20 lat jest silnie zaangażowana w walkę z seksualnym niewolnictwem w Indiach. Przedstawia okoliczności, w jakich doszło do rozpoczęcia jej działalności, inicjatywy przez nią podejmowane oraz rezultaty działań. W skrótowej formie prezentuje opowieści, które zebrała Krishnan spotykając się z pokrzywdzonymi kobietami i dziećmi. Autorka podejmuje również próbę przedstawienia hinduskich modeli kobiecości i wykazania jaki mają one wpływ na obecną sytuację kobiet. Tekst oparty jest w dużej mierze na źródłach internetowych, artykułach publikowanych na portalach informacyjnych, wywiadach przeprowadzonych z Krishnan oraz zapisie konferencji z jej udziałem.Udostępnienie publikacji Wydawnictwa Uniwersytetu Łódzkiego finansowane w ramach projektu „Doskonałość naukowa kluczem do doskonałości kształcenia”. Projekt realizowany jest ze środków Europejskiego Funduszu Społecznego w ramach Programu Operacyjnego Wiedza Edukacja Rozwój; nr umowy: POWER.03.05.00-00-Z092/17-00
Integrin Regulators in Neutrophils
No full textNeutrophils are the most abundant leukocytes in humans and are critical for innate immunity and inflammation. Integrins are critical for neutrophil functions, especially for their recruitment to sites of inflammation or infections. Integrin conformational changes during activation have been heavily investigated but are still not fully understood. Many regulators, such as talin, Rap1-interacting adaptor molecule (RIAM), Rap1, and kindlin, are critical for integrin activation and might be potential targets for integrin-regulating drugs in treating inflammatory diseases. In this review, we outline integrin activation regulators in neutrophils with a focus on the above critical regulators, as well as newly discovered modulators that are involved in integrin activation
Antioxidative properties of fermented poultry green bones
No full textThis Dissertation / Report is the outcome of investigation carried out by the creator(s) / author(s) at the department/division of Central Food Technological Research Institute (CFTRI), Mysore mentioned below in this page
Neuronal plasticity of human Wharton's jelly mesenchymal stromal cells to the dopaminergic cell type compared with human bone marrow mesenchymal stromal cells
No full textA Study on Abstractive Summarization Techniques in Indian Languages
No full textAbstractNatural Language Processing is a vast area which has great importance when people started to interpret human language from one form to another. Summarization is one of the research works in NLP which concentrates on providing meaningful summary using various NLP tools and techniques. Since huge amount of information is used across the digital world, it is highly essential to have automatic summarization techniques. Extractive and Abstractive summarization are the two summarization techniques available. A lot of research works are being carried out in this area especially in extractive summarization. Even though more works are carried out using extractive method, meaningful summary can be attained using abstractive summary techniques which make it more complex. In Indian languages, very few works are carried out in the field of abstractive summarization and there is high need for having research works being carried out in this area. Here, we are concentrating on the various techniques available for abstractive summarization and also try to explain the limited works currently available in abstractive summary field of Indian languages
A Study on Byzantine Fault Tolerance Methods in Distributed Networks
No full textAbstractWith the evolution of new technologies, the growing reliance on networks has increased immensely. Such an advancement induces malicious attacks and software errors to occur frequently. Building robust network services that can tolerate a wide range of failure types is a fundamental problem in distributed systems. The most fundamental approach, called Byzantine fault tolerance, helps mask arbitrary failures exhibited by failing nodes. Here, the problem of reliably broadcasting messages in a mutli hop network is dealt, where some nodes are likely to fail. Most of the existing solutions require high network connectivity and these requirements become difficult to satisfy when the network grows large. In this paper, a study on various Byzantine fault tolerant methods has been carried out which has been developed and implemented by research experts in this field. These solutions are particularly customized to adapt to sparsely connected networks
