scholarly journals Temperature-induced reorganisation of Schistocephalus solidus (Cestoda) proteome during the transition to the warm-blooded host

Biology Open ◽  
2021 ◽  
Vol 10 (11) ◽  
Author(s):  
Ekaterina V. Borvinskaya ◽  
Albina A. Kochneva ◽  
Polina B. Drozdova ◽  
Olga V. Balan ◽  
Victor G. Zgoda
Keyword(s):  
Model Experiment ◽  
Protein Production ◽  
Culture Medium ◽  
Gasterosteus Aculeatus ◽  
Protein Composition ◽  
Tegument Protein ◽  
Schistocephalus Solidus ◽  
Induced Genes

ABSTRACT The protein composition of the cestode Schistocephalus solidus was measured in an experiment simulating the trophic transmission of the parasite from a cold-blooded to a warm-blooded host. The first hour of host colonisation was studied in a model experiment, in which sticklebacks Gasterosteus aculeatus infected with S. solidus were heated at 40°C for 1 h. As a result, a decrease in the content of one tegument protein was detected in the plerocercoids of S. solidus. Sexual maturation of the parasites was initiated in an experiment where S. solidus larvae were taken from fish and cultured in vitro at 40°C for 48 h. Temperature-independent changes in the parasite proteome were investigated by incubating plerocercoids at 22°C for 48 h in culture medium. Analysis of the proteome allowed us to distinguish the temperature-induced genes of S. solidus, as well as to specify the molecular markers of the plerocercoid and adult worms. The main conclusion of the study is that the key enzymes of long-term metabolic changes (glycogen consumption, protein production, etc.) in parasites during colonisation of a warm-blooded host are induced by temperature.

scholarly journals Temperature-induced reorganisation of Schistocephalus solidus (Cestoda) proteome during the transition to the warm-blooded host

2021 ◽  
Author(s):  
Ekaterina Borvinskaya ◽  
Albina Kochneva ◽  
Polina Drozdova ◽  
Olga Balan ◽  
Victor Zgoda
Keyword(s):  
Ribosomal Proteins ◽  
Gasterosteus Aculeatus ◽  
Protein Composition ◽  
Short Term ◽  
Initiation Factors ◽  
Schistocephalus Solidus ◽  
Translation Initiation Factors ◽  
Short Term Experiment

The protein composition (proteome) of cestode Schistocephalus solidus was measured in an experiment simulating the transition of the parasite from a cold-blooded to a warm-blooded host. Infective S. solidus plerocercoids obtained from the three-spined stickleback Gasterosteus aculeatus were heated at 40 °C for 1 hour or cultured in vitro at 40 °C and 22 °C for 48 hours. In short-term experiment, the content of only one tegument protein was evidenced to decrease after heating. After long-term heating, which triggered parasite sexual maturation, an increase in the content of ribosomal proteins, translation initiation factors and enzymes of the amino acid biosynthesis pathway was observed. The synthesis of certain gene products for carbohydrate metabolism, including glycolysis/gluconeogenesis, was found to be regulated in parasite by temperature.

scholarly journals The three-spined stickleback-Schistocephalus solidussystem: an experimental model for investigating host-parasite interactions in fish

Parasitology ◽  
2009 ◽  
Vol 137 (3) ◽  
pp. 411-424 ◽  
Author(s):  
I. BARBER ◽  
J. P. SCHARSACK
Keyword(s):  
Immune Responses ◽  
Gasterosteus Aculeatus ◽  
Laboratory Model ◽  
Host Fish ◽  
Schistocephalus Solidus ◽  
Host Parasite Interactions ◽  
Experimental Laboratory ◽  
Host Parasite ◽  
Host Behaviour

SUMMARYPlerocercoids of the pseudophyllidean cestodeSchistocephalus solidusinfect the three-spined sticklebackGasterosteus aculeatus, with important consequences for the biology of host fish. Techniques for culturing the parasitein vitroand generating infective stages that can be used to infect sticklebacks experimentally have been developed, and the system is increasingly used as a laboratory model for investigating aspects of host-parasite interactions. Recent experimental laboratory studies have focused on the immune responses of hosts to infection, the consequences of infection for the growth and reproductive development of host fish and the effects of infection on host behaviour. Here we introduce the host and the parasite, review the major findings of these recent experimental infection studies and identify further aspects of host parasite interactions that might be investigated using the system.

scholarly journals Long-term in Vitro Regeneration of Hamelia patens

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 873G-874
Author(s):  
D. Sankhla ◽  
T.D. Davis ◽  
N. Sankhla ◽  
A. Upadhyaya
Keyword(s):  
Culture Medium ◽  
In Vitro Regeneration ◽  
Shoot Formation ◽  
Shoot Tips ◽  
Shoot Cultures ◽  
Ms Medium ◽  
Prolific Shoot ◽  
Ornamental Shrub

This report describes an efficient in vitro regeneration protocol for H. patens (firebush), a heat-tolerant ornamental shrub native to tropical and subtropical America. Shoot cultures were initially established using shoot tips placed on MS-revised medium containing 2.3 μM 2,4-D, 2.3 μM kinetin, and 0.25% polyvinylpyrrolidone. Other types of explants (nodal and internodal segments, leaf pieces, floral buds) did not regenerate shoots when placed on this medium. Two-month-old plantlets derived from the shoot tips were subcultured on MS medium supplemented with 0.5 μM thidiazuron (TDZ), and within 3 to 4 weeks, some callus was produced at the root–shoot junction. When this callus, with a small portion of the root and shoots, was placed on MS medium with 0.05 μM TDZ and 0.01 μM ABA, prolific shoot formation occurred within 3 to 4 weeks followed by root formation. By regular subculturing every 5 to 6 weeks, hundreds of plantlets have been obtained over the past 3 years with no apparent decline in regeneration potential. Addition of activated charcoal (0.5%) to the culture medium has greatly improved growth of the plantlets.

scholarly journals Expression of Otx Genes in Müller Cells Using an In Vitro Experimental Model of Retinal Hypoxia

2021 ◽  
Vol 2021 ◽  
pp. 1-10
Author(s):  
Claudio Azzolini ◽  
Simone Donati ◽  
Giovanni Micheloni ◽  
Vittoria Moretti ◽  
Roberto Valli ◽  
...  
Keyword(s):  
Recovery Time ◽  
Culture Medium ◽  
Cobalt Chloride ◽  
Retinal Diseases ◽  
Cellular Events ◽  
In Vitro Response ◽  
Retinal Pathologies ◽  
Eye Morphogenesis

Introduction. Müller glial cells typically activate to react to hypoxic tissue damage in several retinal diseases. We evaluated the in vitro response to a hypoxia-mimicking stimulus on the expression of a set of genes, known to contribute to eye morphogenesis and cell differentiation. Materials and Methods. A MIO-M1 Müller cell line was cultured in a hypoxia-mimicking environment by the addition of cobalt chloride to the culture medium, followed by a recovery time in which we mimic restoration from the hypoxic insult. The HIF-1α protein and VEGF-A gene expression were quantified to verify the induction of a hypoxia-like state. Results. Among the genes under study, we did not observe any difference in the expression levels of Otx1 and Otx2 during treatment; conversely, Otx1 was overexpressed during recovery steps. The VEGF-A gene was strongly upregulated at both the CoCl2 and recovery time points. The transactivated isoform (TA) of the TP73 gene showed an overexpression in long-term exposure to the hypoxic stimulus with a further increase after recovery. Discussion. Our molecular analysis is able to describe the activation of a set of genes, never before described, that can drive the response to a hypoxia-like status. The improved comprehension of these cellular events will be useful for designing new therapeutical approaches for retinal pathologies.

AMINO ACIDS AND PROTEINS IN CULTURE FILTRATES OF THE ANTHRACNOSE PATHOGEN FUNGUS COLLETOTRICHUM LINI

2020 ◽  
pp. 121-127
Author(s):  
N. V. Proletova ◽  
Keyword(s):  
Amino Acids ◽  
Culture Filtrate ◽  
Nutrient Medium ◽  
Culture Medium ◽  
Protein Composition ◽  
Entire Period ◽  
Culture Filtrates ◽  
Russian Research Institute ◽  
Russian Research

The research was carried out on the basis of laboratory biotechnologies of All-Russian research institute of flax (Tver region) in 2010–2012, 2016. The aim of the work was to determine the amino acid and protein composition of culture filtrates of the anthracnose pathogen fungus Colletotrichum lini Manns et Bolley in order to adjust the concentration of selective agent in the nutrient medium when creating in vitro new flax genotypes resistant to anthracnose. It was established that the culture filtrates of strains 527 and 608 contain such amino acids as alanine, glycine, asparagine, cysteine, threonine, aspartic acid, glutamic acid, as well as arginine in strain 527 and traces of tyrosine and lysine in strain 608. It was established that the concentration of amino acids in EC of strain 527 was significantly higher than in culture filtrate of strain 608. It was shown that the toxicity of the culture filtrate depended on the degree of aggressiveness of the anthracnose pathogen strain – culture filtrate of a strongly aggressive strain is more toxic than the culture filtrate of a weakly aggressive strain. Studies have revealed that when cultivating the fungus-causative agent of anthracnose on a nutrient medium, as the mycelium of fungus grew, the concentrations of asparagine, alanine, aspartic and glutamic acids, and glycine decreased in the culture filtrates. It was established that the change in amount of proteins happened during the entire period of cultivation of the mycelium of fungus on a liquid nutrient medium. It is shown that accumulation and content of proteins in culture filtrates of strains of different aggressiveness occurs in different ways. The more aggressive strain is (639), which is more toxic, contains and accumulates more proteins in the culture medium during the entire period of growth and development the less aggressive strain is (419).

scholarly journals Extract of Amburana cearensis maintains the survival of ovine preantral follicles during long-term ovarian tissue transport and promotes primordial follicle activation after in vitro culture

2018 ◽  
Vol 39 (5) ◽  
pp. 2001
Author(s):  
Vanúzia Gonçalves Menezes ◽  
Ricássio De Sousa Barberino ◽  
Bruna Bortoloni Gouveia ◽  
Rodrigo José de Souza Gonçalves ◽  
Jackson Roberto Guedes da Silva Almeida ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Culture Medium ◽  
Ovarian Tissue ◽  
Primordial Follicle ◽  
Chi Square ◽  
Preantral Follicles ◽  
Primordial Follicle Activation ◽  
Chi Square Test

This study evaluated the effect of Amburana cearensis extract as a preservation or culture medium for ovine ovarian tissue. Ovarian fragments were fixed in 4% buffered formaldehyde for 18 h (fresh control), stored in Minimal Essential Medium (MEM) or in A. cearensis extract (0.1; 0.2 or 0.4 mg/mL) at a temperature of 4ºC for 6, 12 or 24 h (preservation - experiment 1) or cultured for 7 days in ?-MEM+ or in A. cearensis extract without (0.1; 0.2 or 0.4 mg/mL) or with supplements (0.1+ ; 0.2+ or 0.4+ mg/ mL; experiment 2). The percentages of morphologically normal follicles and follicular activation were submitted to analysis of variance (ANOVA) and Tukey´s test. The values of TUNEL-positive cells were submitted to Chi-square test (P < 0.05). The storage of fragments for 6 h in MEM showed higher percentages of normal follicles (62%) and a lower rate of TUNEL positive cells (36.17%) compared to other treatments (normal follicles: 46%; 43% and 52%; TUNEL positive cells: 58.57%; 55.30% and 55.63% for Amb 0.1; Amb 0.2 and Amb 0.4 mg/mL, respectively). However, after 12 or 24 h, MEM (12 h: 48%; 24 h: 45%) and Amb 0.2 mg/mL (12 h: 37%; 24 h: 39%) showed similar percentages of normal follicles and TUNEL positive cells (MEM - 12 h: 43.26%; 24 h: 58%; Amb 0.2 mg/mL - 12 h: 50%; 24 h: 61%). After culture, ?-MEM+ recorded a higher percentage of normal follicles (58.25%) than A. cearensis treatments (32.8%; 25.4% and 34.2% for Amb 0.1; Amb 0.2 and Amb 0.4 mg/mL, and 22.25%; 20.0% and 36.6% for Amb 0.1+ ; Amb 0.2+ and Amb 0.4+ mg/mL, respectively) (P < 0.05). Follicular activation increased in all treatments (52.5%; 36.73%; 54.05%; 47.5% and 58.19% for ?-MEM+ ; Amb 0.1; Amb 0.1+ ; Amb 0.2+ and Amb 0.4+ mg/mL, respectively) compared to the fresh control (11.65%), except for Amb 0.2 mg/mL (23.69%) and Amb 0.4 mg/mL (28.85%) (P > 0.05). Moreover, after in vitro culture, A. cearensis at a concentration of 0.1 mg/mL maintained the percentage of TUNEL positive cells (30.0%) in a way that is similar to that observed in the fresh control (22%) (P > 0.05). In conclusion, ovine preantral follicles can be preserved at 4°C in MEM for 6 h. For longer periods of preservation (24 h), MEM and 0.2 mg/mL A. cearensis are recommended. Moreover, after in vitro culture, A. cearensis extract (0.1 mg/mL) showed higher activation and lower DNA fragmentation in ovine preantral follicles.

scholarly journals Effects of Exposure to Tobacco Cigarette, Electronic Cigarette and Heated Tobacco Product on Adipocyte Survival and Differentiation In Vitro

Toxics ◽  
2020 ◽  
Vol 8 (1) ◽  
pp. 9 ◽  
Author(s):  
Zoi Zagoriti ◽  
Mohamed A. El Mubarak ◽  
Konstantinos Farsalinos ◽  
Stavros Topouzis
Keyword(s):  
Culture Medium ◽  
Electronic Cigarettes ◽  
Body Weight Loss ◽  
Tobacco Product ◽  
Dependent Manner ◽  
Ppar Γ ◽  
Beige Adipocytes ◽  
Thermogenic Adipocytes

Cigarette smoking (CS) causes significant morbidity worldwide, attributed to the numerous toxicants generated by tobacco combustion. Electronic cigarettes (ECIG) and heated tobacco products (HTP) are considered alternative smoking/vaping products that deliver nicotine through an inhaled aerosol and emit fewer harmful constituents than CS. However, their long-term impacts on human health are not well established. Nicotine exposure has been linked to lipolysis and body weight loss, while smoking has been associated with insulin resistance and hyperinsulinemia. Enhanced function of beige (thermogenic) adipocytes has been proposed as a means to reduce obesity and metabolic disorders. In this study, we compared the effect of extract-enriched media via exposure of culture medium to CS, HTP aerosol, and ECIG aerosol on the viability and the differentiation of 3T3-L1 pre-adipocytes to beige adipocytes. Only CS extract caused a decrease in cell viability in a dose- and time-dependent manner. Furthermore, relative lipid accumulation and expression levels of the adipocyte markers Pgc-1α, Ppar-γ and Resistin were significantly decreased in cells exposed to CS extract. Our results demonstrate that CS extract, in contrast to HTP and ECIG extracts, significantly impairs differentiation of pre-adipocytes to beige adipocytes and may therefore impact significantly adipose tissue metabolic function.

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.

Effect of phosphatidylcholine–cholesterol liposomes on Entamoeba histolytica virulence

2010 ◽  
Vol 56 (12) ◽  
pp. 987-995 ◽  
Author(s):  
Jesús Serrano-Luna ◽  
Manuel Gutiérrez-Meza ◽  
Ricardo Mejía-Zepeda ◽  
Silvia Galindo-Gómez ◽  
Víctor Tsutsumi ◽  
...  
Keyword(s):  
Entamoeba Histolytica ◽  
Culture Medium ◽  
In Vitro Cultivation ◽  
Biological Functions ◽  
Axenic Cultures ◽  
Term Maintenance ◽  
Histolytica Strain

Trophozoites of Entamoeba histolytica HM-1:IMSS become less virulent after long-term maintenance in axenic cultures. The factors responsible for the loss of virulence during in vitro cultivation remain unclear. However, it is known that in vitro cultivation of amoeba in culture medium supplemented with cholesterol restores their virulence. In this study, we analyzed the effect of adding phosphatidylcholine–cholesterol (PC–Chol) liposomes to the culture medium and evaluated the effect of this lipid on various biochemical and biological functions of E. histolytica HM-1:IMSS in terms of its virulence. The addition of PC–Chol liposomes to the culture medium maintained the virulence of these parasites against hamster liver at the same level as the original virulent E. histolytica strain, even though these amoebae were maintained without passage through hamster liver for 18 months. The trophozoites also showed increased endocytosis, erythrophagocytosis, and carbohydrate residue expression on the amoebic surface. Protease activities were also modified by the presence of cholesterol in the culture medium. These findings indicate the capacity of cholesterol to preserve amoeba virulence and provide an alternative method for the maintenance of virulent E. histolytica trophozoites without the need for in vivo procedures.

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