Immunocytochemistrical Studies on Cytoskeletal Change in Cultured Cardiomyocyte of Tilapia

2000 ◽  
Vol 6 (S2) ◽  
pp. 888-889
Author(s):  
L. C. Tung
Keyword(s):  
Culture Medium ◽  
Cultured Cells ◽  
Calf Serum ◽  
Amino Acid Mixture ◽  
Penicillin G ◽  
Acid Mixture ◽  
Minimal Essential Medium ◽  
Cultured Fish ◽  
Cytoskeletal Change

In the present study, the myofibril regeneration in the long-term cultured fish cardiomyocytes was studied with immunocytochemistry.Adult Tilapia heart was dissociated into a single-cell suspension with collagenase and protease-minced tissue method. The culture medium was Eagle's minimal essential medium (MEM) with Earle's salts, supplemented with 10% fetal calf serum, 1 x nonessential amino acid mixture, 100 IU/ml penicillin G, and 100 μg/ml streptomycin. The cultured cells were grown in a humidified CO2 incubator at 28°Cand in a medium without glutamine for eliminating fibroblast contamination. In the initial 24 h culture, the elongated-shape cells gradually shortened from their both ends and rounded up. Over 5 to 6 days postcultivation, the cells attached to the bottom of the culture flask and began to protrude pseudopodia. The cells could not be subcultured and also proliferated indefinitely. The life span of cells in culture was 30 to 60 days.

scholarly journals Emerging trends in management of propionic acidemia

2014 ◽  
Vol 58 (3) ◽  
pp. 237-242 ◽  
Author(s):  
Muhammad Rafique
Keyword(s):  
Sodium Benzoate ◽  
Protein Intake ◽  
Propionic Acidemia ◽  
Amino Acid Mixture ◽  
Fresh Frozen Plasma ◽  
Acid Mixture ◽  
Fresh Frozen ◽  
Almost All ◽  
Term Management

Objetivo : To evaluate the therapeutic agents used during metabolic crises and in long-term management of patients with propionic acidemia (PA).Materials and methods : The records of PA patients were retrospectively evaluated.Results : The study group consisted of 30 patients with 141 admissions. During metabolic crises, hyperammonemia was found in 130 (92%) admissions and almost all patients were managed with normal saline, ≥ 10% dextrose, and restriction of protein intake. In 56 (40%) admissions, management was done in intensive care unit, 31 (22%) with mechanical ventilation, 10 (7%) with haemodialysis, 16 (11%) with vasopressor agents, and 12 (9%) with insulin. In the rescue procedure, L-carnitine was used in 135 (96%) patients, sodium bicarbonate in 116 (82%), sodium benzoate in 76 (54%), and metronidazole in 10 (7%), biotin in about one-quarter, L-arginine in one third, and antibiotics in three-quarter of the admissions. Blood/packed RBCs were used in 28 (20%) patients, platelets in 26 (18%), fresh frozen plasma in 8 (6%), and granulocyte-colony stimulating factors in 10 (7%) admissions. All patients were managed completely/partially with medical nutrition formula plus amino acid mixture, vitamins and minerals. For long-term management 24 (80%) patients were on L-carnitine, 22 (73%) on sodium benzoate, 6 (20%) on biotin, one half on alkaline therapy and 4 (13%) on regular metronidazole use. Almost all patients were on medical formula and regular follow-up.Conclusion : Aggressive and adequate management of acute metabolic crises with restriction of protein intake, stabilization of patient, reversal of catabolism, and removal of toxic metabolites are essential steps. Concerted efforts to ensure adequate nutrition, to minimize the risk of acute decompensation and additional therapeutic advances are imperative to improve the outcome of PA patients. Arq Bras Endocrinol Metab. 2014;58(3):237-42

The effect of osmolarity on mouse parthenogenesis

Development ◽  
1974 ◽  
Vol 31 (2) ◽  
pp. 513-526
Author(s):  
M. H. Kaufman ◽  
M. A. H. Surani
Keyword(s):  
Protein Synthesis ◽  
Culture Medium ◽  
Culture Media ◽  
Polar Body ◽  
Amino Acid Mixture ◽  
Culture Conditions ◽  
Follicle Cells ◽  
Acid Mixture ◽  
Second Polar Body

Eggs from (C57B1 × A2G)F1 mice were activated by treatment with hyaluronidase, which removed the follicle cells, and cultured in vitro. Observations were made 6–8 h after hyaluronidase treatment to determine the frequency of activation and the types of parthenogenones induced. Cumulus-free eggs resulting from hyaluronidase treatment were incubated for 2¼ h in culture media of various osmolarities. The frequency of activation was found to be dependent on the postovulatory age of oocytes, while the types of parthenogenones induced were dependent on the osmolarity of the in vitro culture medium and their postovulatory age. Culture in low osmolar medium suppressed the extrusion of the second polar body (2PB). This decreased the incidence of haploid eggs with a single pronucleus and 2PB and immediately cleaved eggs from 97·5% to 42·3% of the activated population. Where 2PB extrusion had been suppressed, 97·4% of parthenogenones contained two haploid pronuclei. Very few were observed with a single and presumably diploid pronucleus. Serial observations from 11 to 18 h after hyaluronidase treatment were made on populations of activated eggs as they entered the first cleavage mitosis after 2¼ h incubation in medium either of normal (0·287 osmol) or low (0·168 osmol) osmolarity. A delay in the time of entry into the first cleavage mitosis similar to the duration of incubation in low osmolar medium was observed. Further, eggs were incubated in control and low osmolar culture media containing uniformly labelled [U-14C]amino acid mixture to examine the extent of protein synthesis in recently activated eggs subjected to these culture conditions. An hypothesis is presented to explain the effect of incubation in low osmolar culture medium in delaying the first cleavage mitosis.

Megakaryocytic Progenitors in Long Term Cultures from MDS Patients.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4924-4924
Author(s):  
Maria Juliana Majado ◽  
María I. Macizo ◽  
Consuelo González-García ◽  
Eduardo Salido ◽  
José A. Molina ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Culture Medium ◽  
Horse Serum ◽  
Calf Serum ◽  
Methyl Cellulose ◽  
Refractory Anemia ◽  
Foetal Calf Serum ◽  
Cell Counts ◽  
Control Bone

Abstract Introduction: bone marrow long-term cultures (LTC) have been reported to produce very poor adherent stromal layer in MDS, but there are not many studies about the behaviour of the progenitor cells in supernatant culture medium. In this study we report the results of megakaryocytic progenitors (CFU-MK) in the supernatant of LTC of patients with refractory anemia (RA). Material and method: LTCs were performed to 11 RA patients and to 13 normal multiorgan donors as control. Bone marrow red cells were sedimented with 1% methyl-cellulose and seeded for LTC in 10ml flasks (Nalge Nunc International), with Iscover medium supplemented with horse serum, foetal calf serum, hydrocortisone and carbonic air, at a final concentration of 1x106 per ml. Flasks were placed in an incubator for 8 weeks. Half culture medium volume was renovated weekly, cell counts and assays of CFU-MK (Megacult C, Stem Cell Technologies), were performed. CFU-MK colonies were separated in three groups, containing 3–20 cells, 21–50 cells and more than 50 cells. Student t-test was used for statistical comparisons. Results: Are in the following table, expressed as mean + standard desviation. After the third week no colony growth was observed in normal such as in MDS. Growth of CFU-MK colonies containing more than 50 cells was higher in basal control bone marrow, no statistical differences were found in the rest of the cultures. Results CFU-MK(3–30 Cells) CFU-MK(20–50 Cells) CFU-MK (>50 Cells) CFU-MK total w: week Control basal 15.75±15.79 3.33±5.42 4.88±7.44 22.42±26.43 RA basal 7.85±8.66 0.80±1.01 0.70±0.98 9.45±9.46 t-St ns ns 0.047 ns Control w 1 3.54±4.42 0.7±1.44 0.74±1.48 4.98±6.62 RA w 1 3.29±4.52 0.29±0.93 0.25±0.67 3.82±5.65 t-St ns ns ns ns Control w 2 0.55±1.14 0.07±0.24 0.05±0.25 0.67±1.43 RA w 2 0.18±0.32 0.00±0.00 0.04±0.13 0.21±0.38 t-St ns ns ns ns Control w 3 0.18±0.53 0.00±0.00 0.00±0.00 0.18±0.53 RA w 3 0.00±0.21 0.00±0.00 0.00±0.00 0.00±0.21 t-St ns ns Discussion: No important difference was found in LTC supernatant CFU-MK in our RA patients, and this supports the idea that the stromal damage is more important, in the pathophysyology of MDS, than that of the stem cells.

Long-term survival and aldosterone secretion pattern of aldosteronoma in culture

1983 ◽  
Vol 104 (4) ◽  
pp. 495-501 ◽  
Author(s):  
Tetsuro Okabe ◽  
Hiroshi Hidaka ◽  
Nakaaki Ohsawa ◽  
Toshio Tsushima
Keyword(s):  
Angiotensin Ii ◽  
Primary Aldosteronism ◽  
Culture Medium ◽  
Cultured Cells ◽  
Aldosterone Secretion ◽  
Term Survival ◽  
Eosinophilic Cytoplasm

Abstract. In an attempt to obtain an in vitro experimental model for aldosteronoma, primary culture was initiated with adenomas from 3 patients with primary aldosteronism. The cells grown in culture retained the morphology and functional properties characteristic of aldosteronoma cells well for periods of up to 200 days. The cells formed monolayer cell colonies and showed an epithelioid morphology with small nuclei containing prominent nucleoli. The cells possessed a clear, eosinophilic cytoplasm resembling that of aldosteronoma cells in vivo. The cultured cells continued to secrete large amounts of aldosterone throughout the culture period. The cells responded to angiotensin II and III by increased release of aldosterone into the culture medium. They also responded to Db-cAMP and ACTH by increased secretion of the hormone.

A Serum-Free in vitro Culture System for Crayfish Organs

1986 ◽  
Vol 41 (4) ◽  
pp. 472-476 ◽  
Author(s):  
Gerd Gellissen ◽  
Marco Traub ◽  
Klaus-Dieter Spindler
Keyword(s):  
Culture System ◽  
Amino Acid Mixture ◽  
Acid Mixture ◽  
Serum Free Medium ◽  
Free Medium ◽  
Astacus Leptodactylus ◽  
Serum Free ◽  
In Vitro Culture System

Midgut gland and hypodermis of the crayfish Astacus leptodactylus have been cultured in a serum-free medium for several days. The medium consists of 1 part of van Harreveld solution and 1 part of an amino acid mixture supplemented with 1.2 mᴍ Na2HP04, 12 mᴍ Hepes and 80 mᴍ glucose. The antibiotics penicillin (15 mg/l) and streptomycin (25 mg/1) were added for long term culturing. This medium, called TG medium, allows the maintainance of the tissues for more than 100 h without any loss of their viability with respect to protein synthesis and secretion.

scholarly journals Minocycline Protects Dopaminergic Neurons Against Long-Term Rotenone Toxicity

2010 ◽  
Vol 37 (1) ◽  
pp. 81-85 ◽  
Author(s):  
Khaled Radad ◽  
Rudolf Moldzio ◽  
Wolf-Dieter Rausch
Keyword(s):  
Lactate Dehydrogenase ◽  
Tyrosine Hydroxylase ◽  
Cell Cultures ◽  
Culture Medium ◽  
Cultured Cells ◽  
Disease Process ◽  
Dopaminergic Cell ◽  
20 Nm

Background:In Parkinson's disease, most of current therapies only provide symptomatic treatment and so far there is no drug which directly affects the disease process.Objectives:To investigate the neuroprotective effects of minocycline against long-term rotenone toxicity in primary dopaminergic cell cultures.Methods:Embryonic mice of 14-days-old were used for preparation of primary dopaminergic cell cultures. On the 6th day in vitro, prepared cultures were treated both with minocycline alone (1, 5, 10 and 20 μM) and concomitantly with rotenone (5 and 20 nM) and minocycline. Cultures were incubated at 37°C for six consecutive days. On Day 12 in vitro culture medium was aspirated and used for measuring lactate dehydrogenase. Cultured cells were fixed in 4% paraformaldhyde and stained immunohistochemically against tyrosine hydroxylase.Results:Treatment of cultures with 5 and 20 nM of rotenone significantly decreased the survival of tyrosine hydroxylase immunoreactive neurons by 27 and 31% and increased the release of lactate dehydrogenase into the culture medium by 31 and 236%, respectively compared to untreated controls. Minocycline (1, 5, 10 μM) significantly protected tyrosine hydroxylase immunoreactive neurons by 17, 15 and 19% and 13, 22 and 23% against 5 and 20 nM of rotenone, respectively compared to rotenone-treated cultures. Minocycline (only at 10 μM) significantly decreased the release of lactate dehydrogenase by 79% and 133% against 5 and 20 nM of rotenone, respectively.Conclusion:Minocycline has neuroprotective potential against the progressive loss of tyrosine hydroxylase immunoreactive neurons induced by long-term rotenone toxicity in primary dopaminergic cultures.

A potassium permanganate fixative for membranes in sections

1990 ◽  
Vol 48 (3) ◽  
pp. 722-723
Author(s):  
Jindan Song
Keyword(s):  
Potassium Permanganate ◽  
Cultured Cells ◽  
Calf Serum ◽  
Trisodium Citrate ◽  
Membrane System ◽  
Adenocarcinoma Cell Line ◽  
Mouse Embryo Fibroblast ◽  
African Green Monkey Kidney ◽  
Adenocarcinoma Cell ◽  
Green Monkey

Potassium permanganate has been used as a fixative for the botanical specimen and membrane system in thin section by Glauert (1975). A new potassium permanganate fixative ( Trisodium citrate 60mM, Potassium chloride 25mM, Magnesium chloride 35mM, and Potassium permanganate 125mM ) for localizing membranous system in whole_mount cultured cells with standard trasmission electron microscopy and phase_contrast microscopy has been developed). Here, we report that using this new potassium permanganate fixative for membranous system in sections.Cultured cells, CV_1 (African green monkey kidney epithelial cells), Balb/c 3T3 ( Mouse embryo fibroblast ) and MCF_7 (Human adenocarcinoma cell line) were used for this study. All cells were grown on 35mm plastic dishes in DME medium containing 5% calf serum at 37 c with 100% humidity and 5% CO2. Using the potassium permanganate fixative to fix the cells for about 7 minutes. After fixation, the cells were dehydrated in a graded series of ethanol.

An open-label, single-center pilot study to test the effects of an amino acid mixture in older patients admitted to internal medicine wards

Nutrition ◽  
2020 ◽  
Vol 69 ◽  
pp. 110588 ◽  
Author(s):  
Francesco Bellanti ◽  
Aurelio Lo Buglio ◽  
Elena Di Stasio ◽  
Giorgia di Bello ◽  
Rosanna Tamborra ◽  
...  
Keyword(s):  
Internal Medicine ◽  
Amino Acid ◽  
Pilot Study ◽  
Older Patients ◽  
Amino Acid Mixture ◽  
Acid Mixture ◽  
Single Center ◽  
Open Label ◽  
Internal Medicine Wards

scholarly journals Structure-Properties Correlation of Cross-Linked Penicillin G Acylase Crystals

Crystals ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 451
Author(s):  
Marta Kubiak ◽  
Janine Mayer ◽  
Ingo Kampen ◽  
Carsten Schilde ◽  
Rebekka Biedendieck
Keyword(s):  
Mechanical Stability ◽  
Structural Characteristics ◽  
Penicillin G ◽  
Bacillus Species ◽  
Diffusion Limitations ◽  
Small Water ◽  
Long Term Stability ◽  
Atomic Force ◽  
First Time

In biocatalytic processes, the use of free enzymes is often limited due to the lack of long-term stability and reusability. To counteract this, enzymes can be crystallized and then immobilized, generating cross-linked enzyme crystals (CLECs). As mechanical stability and activity of CLECs are crucial, different penicillin G acylases (PGAs) from Gram-positive organisms have proven to be promising candidates for industrial production of new semisynthetic antibiotics, which can be crystallized and cross-linked to characterize the resulting CLECs regarding their mechanical and catalytic properties. The greatest hardness and Young’s modulus determined by indentation with an atomic force microscope were observed for CLECs of Bacillus species FJAT-PGA CLECs (26 MPa/1450 MPa), followed by BmPGA (Priestia megaterium PGA, 23 MPa/1170 MPa) and BtPGA CLECs (Bacillus thermotolerans PGA, 11 MPa/614 MPa). In addition, FJAT- and BtPGA CLECs showed up to 20-fold higher volumetric activities compared to BmPGA CLECs. Correlation to structural characteristics indicated that a high solvent content and low number of cross-linking residues might lead to reduced stability. Furthermore, activity seems to be restricted by small water channels due to severe diffusion limitations. To the best of our knowledge, we show for the first time in this study that the entire process chain for the characterization of diverse industrially relevant enzymes can be performed at the microliter scale to discover the most important relationships and limitations.

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