scholarly journals OLIGOMERIC IGA: THE MAJOR COMPONENT OF THE IN VITRO PRIMARY RESPONSE OF MOUSE SPLEEN FRAGMENTS

1973 ◽  
Vol 138 (4) ◽  
pp. 973-988 ◽  
Author(s):  
Ichiro Nakamura ◽  
Alex Ray ◽  
O. Mäkelä
Keyword(s):  
Sucrose Gradient ◽  
Culture Media ◽  
Keyhole Limpet Hemocyanin ◽  
Aminocaproic Acid ◽  
Primary Response ◽  
Mouse Spleen ◽  
Primary Antibody Response ◽  
Hydroxyphenylacetic Acid ◽  
The Media

The primary antibody response elicited from mouse spleen explants by conjugates of the 3-nitro-5-iodo-4-hydroxyphenylacetic acid (NIP) hapten consisted mostly of the IgA class. Poly-L-lysine, pneumococcal polysaccharide Type SIII, keyhole limpet hemocyanin, and sheep erythrocytes were effective carriers in this system, whereas chicken globulin was not. The anti-NIP response against all of the immunogenic conjugates was detectable in culture media 4 days after explantation and immunization, and reached peak titers by 8–10 days. IgA was identified by sucrose gradient velocity centrifugation in conjunction with the use of a class-specific antiserum. The media collected at 4 days contained low titers of IgM antibody, whereas the peak response at 8 days consisted almost entirely of IgA. The primary response IgA secreted by the spleen fragments was characterized as polymeric by its sedimentation rate through a sucrose gradient, and as polyvalent by its drastically greater avidity for NIP14BSA than for free NIP-aminocaproic acid. Its haptenated phage-inactivating activity was abolished by treatment with 0.1 M 2-mercaptoethanol. These experiments indicate that precursor cells existing in the spleen before primary immunization can give rise to production of polymeric IgA.

Biochemical markers for pregnancy in the spent culture medium of in vitro produced bovine embryos

2021 ◽  
Author(s):  
Gabriela de Oliveira Fernandes ◽  
Marcella Pecora Milazzotto ◽  
Andrei Antonioni Guedes Fidelis ◽  
Taynan Stonoga Kawamoto ◽  
Ligiane de Oliveira Leme ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Biochemical Markers ◽  
Culture Medium ◽  
Culture Media ◽  
Ionization Mass ◽  
Metabolic Profiles ◽  
Bovine Embryos ◽  
The Media

Abstract The present study aimed to identify biomarkers to assess the quality of in vitro produced (IVP) bovine embryos in the culture media. IVP embryos on Day (D) 5 of development were transferred to individual drops, where they were maintained for the last 48 h of culture. Thereafter, the medium was collected and the embryos were transferred to the recipients. After pregnancy diagnosis, the media were grouped into the pregnant and nonpregnant groups. The metabolic profiles of the media were analyzed via electrospray ionization mass spectrometry, and the concentrations of pyruvate, lactate, and glutamate were assessed using fluorimetry. The spectrometric profile revealed that the media from embryos from the pregnant group presented a higher signal intensity compared to that of the nonpregnant group; the ions 156.13 Da [M + H]+, 444.33 Da [M + H]+, and 305.97 Da [M + H]+ were identified as biomarkers. Spent culture medium from expanded blastocysts (Bx) that established pregnancy had a greater concentration of pyruvate (p = 0.0174) and lesser concentration of lactate (p = 0.042) than spent culture medium from Bx that did not establish pregnancy. Moreover, pyruvate in the culture media of Bx can predict pregnancy with 90.9% sensitivity and 75% specificity. In conclusion, we identified markers in the culture media that helped in assessing the most viable IVP embryos with a greater potential to establish pregnancy.

scholarly journals Gas Chromatography-Mass Spectrometry (GC-MS) Analysis of Essential Oils from AgNPs and AuNPs Elicited Lavandula angustifolia In Vitro Cultures

Molecules ◽  
2019 ◽  
Vol 24 (3) ◽  
pp. 606 ◽  
Author(s):  
Aneta Wesołowska ◽  
Paula Jadczak ◽  
Danuta Kulpa ◽  
Włodzimierz Przewodowski
Keyword(s):  
Molecular Weight ◽  
Silver Nanoparticles ◽  
Essential Oils ◽  
Culture Media ◽  
In Vitro Cultures ◽  
Gas Chromatography Mass Spectrometry ◽  
Lavandula Angustifolia ◽  
Gold And Silver Nanoparticles ◽  
The Media

The aim of this study was to determine how the addition of gold and silver nanoparticles to culture media affects the composition of essential oils extracted from Lavandula angustifolia propagated on MS media with the addition of 10 and 50 mg·dm−3 of gold (24.2 ± 2.4 nm) and silver (27.5 ± 4.8 nm) nanocolloids. The oil extracted from the lavender tissues propagated on the medium with 10 mg·dm−3 AgNPs (silver nanoparticles) differed the most with respect to the control; oil-10 compounds were not found at all, and 13 others were detected which were not present in the control oil. The addition of AuNPs (gold nanoparticles) and AgNPs to the media resulted in a decrease of lower molecular weight compounds (e.g., α- and β-pinene, camphene, δ-3-carene, p-cymene, 1,8-cineole, trans-pinocarveol, camphoriborneol), which were replaced by those of a higher molecular weight (τ- and α-cadinol 9-cedranone, cadalene, α-bisabolol, cis-14-nor-muurol-5-en-4-one, (E,E)-farnesol).

scholarly journals Altered Affinity Maturation in Primary Response to (4-hydroxy-3-nitrophenyl) Acetyl (NP) after Autologous Reconstitution of Irradiated C57BL/6 Mice

2002 ◽  
Vol 9 (3) ◽  
pp. 119-125
Author(s):  
Carl De Trez ◽  
Annette Van Acker ◽  
Georgette Vansanten ◽  
Jacques Urbain ◽  
Maryse Brait
Keyword(s):  
Immune Responses ◽  
Single Gene ◽  
Gene Segment ◽  
Keyhole Limpet Hemocyanin ◽  
Primary Response ◽  
Affinity Maturation ◽  
Germinal Center Reaction ◽  
High Affinity ◽  
Primary Antibody Response ◽  
Vh Genes

Immune responses developing in irradiated environment are profoundly altered. The memory anti-arsonate response of A/J mice is dominated by a major clonotype encoded by a single gene segment combination called CRIA. In irradiated and autoreconstituted A/J mice, the level of anti-ARS antibodies upon secondary immunization is normal but devoid of CRIA antibodies. The affinity maturation process and the somatic mutation frequency are reduced. Isotype switching and development of germinal centers (GC) are delayed.The primary antibody response of C57BL/6 mice to the hapten (4-hydroxy-3-nitrophenyl) acetyl (NP)-Keyhole Limpet Hemocyanin (KLH) is dominated by antibodies encoded by a family of closely related VH genes associated with the expression of the λ1 light chain.We investigated the anti-NP primary response in irradiated and autoreconstituted C57BL/6 mice. We observed some splenic alterations as previously described in the irradiated A/J model. Germinal center reaction is delayed although the extrafollicular foci appearance is unchanged. Irradiated C57BL/6 mice are able to mount a primary anti-NP response dominated by λ1 positive antibodies but fail to produce high affinity NP-binding IgGl antibodies. Following a second antigenic challenge, irradiated mice develop enlarged GC and foci. Furthermore, higher affinity NP-binding IgG1 antibodies are detected.

99 DIFFERENTIAL GENE REGULATION OF STEROIDOGENIC TRANSCRIPTS AND ESTRADIOL PRODUCTION FOLLOWING IN VITRO PIG EMBRYO ELONGATION IN ALGINATE HYDROGEL THREE-DIMENSIONAL MATRIX

2012 ◽  
Vol 24 (1) ◽  
pp. 162
Author(s):  
J. R. Miles ◽  
C. N. Sargus ◽  
S. A. Plautz ◽  
J. L. Vallet ◽  
A. K. Pannier
Keyword(s):  
Equal Opportunity ◽  
Culture Media ◽  
Morphological Changes ◽  
Three Dimensional ◽  
Differential Gene Regulation ◽  
Alginate Hydrogels ◽  
The Media ◽  
And Control

Between Day 10 and 12 of gestation, the pig embryo elongates from a sphere to a long thin, filament. During this time, the embryo increases the production of oestrogen via an increase in steroidogenic transcripts, which is critical for maternal recognition of pregnancy. To date, attempts to elongate porcine embryos in vitro have been unsuccessful. Therefore, the objective of this study was to utilise alginate hydrogels to establish a culture system that promotes in vitro embryo elongation with a corresponding increase in steroidogenic transcripts and oestradiol production. In 3 replicate collections, White crossbred gilts (n = 15) were bred at Day 0 of the oestrous cycle. At Day 9 of gestation, reproductive tracts were collected and flushed with RPMI-1640 containing antibiotics. Embryos were recovered, grouped according to size and washed with RPMI-1640 containing antibiotics and 10% fetal bovine serum (FBS). Embryos were randomly assigned to be encapsulated using a double encapsulation technique (0.7% sodium alginate and 1.5% calcium chloride solution) or used as controls. Encapsulated and control embryos were cultured for 96 h in CO2 -pretreated RPMI-1640 containing antibiotics and 10% FBS at 38°C, 5% CO2 in air and 100% humidity. Every 24 h, the embryos were imaged and half of the media was replaced. The removed media was stored at –20°C and used to assess oestradiol levels by radioimmunoassay. At the end of culture, a subset of encapsulated and control embryos were snap frozen and used to assess the expression level of steroidogenic transcripts (STAR, CYP11 and CYP19) using quantitative PCR. All data were analysed using general linear model (GLM) procedures for ANOVA. Cell survival, assessed by blastocyst fragmentation and confirmed by live/dead staining in representative embryos, was greater (P = 0.01) for encapsulated embryos (60.1 ± 4.8%) compared with controls (33.3 ± 4.8%). Of encapsulated embryos, 27% had some morphological change (minor flattening and tubal formation) and 14% had significant morphological changes (considerable flattening and tubal formation elongating through the gel), consistent with in vivo embryo elongation. In contrast, the control embryos had no morphological changes observed and remained spherical during culture. The expression levels of STAR, CYP11 and CYP19 were significantly (P < 0.05) greater in encapsulated embryos compared with control embryos. Furthermore, a significant (P < 0.01) time-dependent increase in oestradiol levels in the culture media of encapsulated embryos was identified compared with controls and culture media alone. These results illustrate that cultured pig embryos encapsulated in alginate hydrogels undergo limited morphological changes with increased expression of steroidogenic transcripts and oestrogen production. †USDA is an equal opportunity provider and employer.

Effects of parathyroid hormone on bone acid hydrolases in tissue culture

1968 ◽  
Vol 46 (2) ◽  
pp. 261-267 ◽  
Author(s):  
Susan Tolnai
Keyword(s):  
Parathyroid Hormone ◽  
Acid Phosphatase ◽  
Bone Cells ◽  
Culture Media ◽  
Enzyme Synthesis ◽  
Maximum Activity ◽  
Acid Protease ◽  
Acid Hydrolases ◽  
The Media

Experiments were carried out to assess the activity of acid hydrolases of bone cells under in vitro conditions. Rat and mouse skull bones were kept in vitro for 2 to 6 days in the presence and absence of added parathyroid hormone. The activities of acid protease (cathepsin D), acid phosphatase, and β-glucuronidase were measured in the culture media and in homogenates prepared from the cultured bones. All three enzymes showed increased activity in various degrees in the media when parathyroid hormone (PTE) was present, but increased enzyme synthesis in the homogenates could be detected only in the case of acid protease and acid phosphatase. The acid protease showed maximum activity at pH 3.0–3.2, and the PTE-stimulated increase was time-dependent. Lysozyme incorporated into the culture medium stimulated acid protease activity, while histone had an opposite effect. Cultures kept in incomplete media for a limited period of time also responded to PTE treatment with an increased release of acid hydrolases into the medium.

scholarly journals Proto-oncogenes of the fos/jun family of transcription factors are positive regulators of myeloid differentiation.

1993 ◽  
Vol 13 (2) ◽  
pp. 841-851 ◽  
Author(s):  
K A Lord ◽  
A Abdollahi ◽  
B Hoffman-Liebermann ◽  
D A Liebermann
Keyword(s):  
Transcription Factors ◽  
Leucine Zipper ◽  
Culture Media ◽  
Terminal Differentiation ◽  
Cell Types ◽  
Primary Response ◽  
Myeloid Differentiation ◽  
Active Transcription

The proto-oncogenes c-jun, junB, junD, and c-fos recently have been shown to encode for transcription factors with a leucine zipper that mediates dimerization to constitute active transcription factors; juns were shown to dimerize with each other and with c-fos, whereas fos was shown to dimerize only with juns. After birth, hematopoietic cells of the myeloid lineage, and some other terminally differentiated cell types, express high levels of c-fos. Still, the role of fos/jun transcription factors in normal myelopoiesis or in leukemogenesis has not been established. Recently, c-jun, junB, and junD were identified as myeloid differentiation primary response genes stably expressed following induction of terminal differentiation of myeloblastic leukemia M1 cells. Intriguingly, c-fos, though induced during normal myelopoiesis, was not induced upon M1 differentiation. To gain further insights into the role of fos/jun in normal myelopoiesis and leukemogenicity, M1fos and M1junB cell lines, which constitutively express c-fos and junB, respectively, were established. It was shown that enforced expression of c-fos, and to a lesser extent junB, in M1 cells results in both an increased propensity to differentiate and a reduction in the aggressiveness of the M1 leukemic phenotype. M1fos cells constitutively expressed immediate-early and late genetic markers of differentiated M1 cells. The in vitro differentiation of normal myeloblasts into mature macrophages and granulocytes, as well as the increased propensity of M1fos leukemic myeloblasts to be induced for terminal differentiation, was dramatically impaired with use of c-fos antisense oligomers in the culture media. Taken together, these observations show that the proto-oncogenes which encode for fos/jun transcription factors play important roles in promoting myeloid differentiation. The ability of the M1 leukemic myeloblasts to be induced for terminal differentiation in the absence of apparent fos expression indicates that there is some redundancy among the fos/jun family of transcription factors in promoting myeloid differentiation; however, juns alone cannot completely compensate for the lack of fos. Thus, genetic lesions affecting fos/jun expression may play a role in the development of "preleukemic" myelodysplastic syndromes and their further progression to leukemias.

scholarly journals Urochloa brizantha cv. Marandu presents a better response to in vitro salt stress than other commercial cultivars

2021 ◽  
Vol 17 (4) ◽  
pp. 74-82
Author(s):  
Paula Beatriz Ramos Guimarães ◽  
Mayara de Oliveira Vidotto Figueiredo ◽  
Tiago Benedito dos Santos ◽  
Alessandra Ferreira Ribas
Keyword(s):  
Salt Stress ◽  
Molecular Mechanisms ◽  
Culture Media ◽  
Proline Content ◽  
Quantitative Expression ◽  
Mechanisms Of Adaptation ◽  
The Media ◽  
Urochloa Brizantha ◽  
Biomass Reduction

Urochloa brizantha is the main forage grass to raise cattle in Brazil, but salt stress can reduce yield. Physiological and molecular mechanisms of adaptation to salt stress remain poorly understood in this species. The objective of this study was to evaluate the responses of three cultivars of U. brizantha to in vitro salt stress. Seeds of three cultivars (Piatã, Marandu, and Xaraés) germinated in filter paper and then transferred to growth on culture media in vitro containing 0, 50, 100, and 200 mg L-1 of sodium chloride (NaCl). Biometric parameters and proline content were determined after 28 days. The data were subjected to analysis of variance and the separation of means was performed by the LSD test (p<0.05). Semi-quantitative expression of the Δ1-pyrroline-5-carboxylate synthase (P5CS1) gene was performed. In all cultivars, increase of NaCl concentration in the media affected roots and shoots growth. Xaraes cultivar presented the greater biomass reduction while Marandu cultivar was the least affected. Salt stress increased by approximated 0.6 folds transcription of the P5CS1 gene in all cultivars. However, Marandu cultivar presented a higher proline content and least biomass reduction suggesting a better response to in vitro to salt stress.

scholarly journals In Vitro Effect of Crude Extract from Traganum Nudatum on Glucose-Uptake in Liver Slices Isolated from Westar Rats

2020 ◽  
Vol 4 (2) ◽  
pp. 550-551
Author(s):  
Faiza Mouderas
Keyword(s):  
Glucose Uptake ◽  
Crude Extract ◽  
Wistar Rats ◽  
Culture Media ◽  
Chronic Hyperglycemia ◽  
Liver Slices ◽  
Insulin Sensitizers ◽  
The Media ◽  
Vitro Effect

Background: Diabetes mellitus is a metabolic disorder characterized by chronic hyperglycemia resulting from defects in insulin secretion, insulin action, or both. There are many classes of drugs used for treatment, and these include insulin sensitizers, insulin secretagogues, and agents that delay the absorption of carbohydrates from the bowel. This study intends to investigate the effect of crude extract from a plant from South Algeria Traganum nudatum (Chenopodiaceae) on glucose uptake in liver slices isolated from Wistar rats. Methods: The liver slices were incubated for 90 min at 37° in normoglycaemic (1g/l of glucose) and hyperglycaemic (3g/l of glucose) KRBA Krebs Ringer Bicarbonate Albumin 4% media using 24 well-polyethylene plates. In each, well different concentrations of insulin (10, 50 and 100µU/ml) and hydromethanolic crude extract (100, 200 and 500µg/ml) were added. After every 30 minutes, aliquots of the culture media were assayed for the determination of glucose left. Results: Tests showed that the glucose left after 90 minutes in the media which contained insulin at 100µg/ml was the lowest (0.44 and 1.41 )g/l in the normo and hyperglycaemic media respectively, which reflect that insulin at this concentration was the most effective on the stimulation of glucose uptake. The extract had the highest effect at 500µg/ml, the concentrations of glucose left after 90 minutes of incubation were found to be (0.38 and 1.31)g/l in the normoglycaemic and hyperglycaemic media respectively. Conclusion: From the obtained results, it can be concluded that our extract seems to have an insulin-like effect on glucose uptake in liver slices isolated from Wistar rats.

scholarly journals In Vitro Multiplication of Potato (Solanum tuberosum L.) cv. Cristal under Different MS Salt and Sucrose Concentrations

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 471E-471
Author(s):  
Gerson R. de L. Fortes ◽  
Luciana B. Andrade ◽  
Marisa de F. Oliveira ◽  
Nilvane T.G. Müller ◽  
Janine T. C. Faria
Keyword(s):  
Culture Media ◽  
Solanum Tuberosum L ◽  
Sucrose Concentration ◽  
Sugar Content ◽  
Potato Cultivar ◽  
In Vitro Multiplication ◽  
Full Strength ◽  
High Dry Matter ◽  
The Media

The potato cultivar Cristal has recently been released by the CPACT/EMBRAPA Breeding Program. Such cultivar was selected for having high dry matter and low sugar content, which makes it desirable for the chip industry. However, this is a recalcitrant cultivar as far as in vitro multiplication is concerned. The aim of this work was to improve the rate of multiplication for this cultivar when it was submitted to different MS salt and sucrose concentrations in the culture media. Two-bud microcuttings were inoculated in test tubes (20 × 150) mm with 10 ml MS media at 3/4-, 1/2-, and full-strength and MS vitamins added to: myo-inositol (100 mg·L–1), agar (7.0 g·L–1) and sucrose as follows: 10, 20 and 30 g·L-1. Each treatment was repeated eight times and each replicate had eight explants. After inoculation the whole material was kept in a growth room at 25 ± 2°C, 16-hr photoperiod and 2000 lux. The evaluation was done 35 days later. It was found and increase in the number of buds as the sucrose concentration in the media decreased. As far as MS salts are concerned no difference in bud number was observed. The rate of multiplication was slightly higher for MS media at full strength and sucrose at low concentration (10 g·L–1). This treatment could be recommended for this cultivar.

scholarly journals Efisiensi Mikropropagasi Pisang Kepok Amorang melalui Modifikasi Formula Media dan Temperatur

2016 ◽  
Vol 6 (2) ◽  
pp. 91
Author(s):  
Yati Supriati
Keyword(s):  
In Vitro Culture ◽  
Incubation Temperature ◽  
Culture Media ◽  
Shoot Multiplication ◽  
Root Induction ◽  
Incubation Condition ◽  
Growth Environment ◽  
Media Formulation ◽  
The Media

<p>Micropropagation Efficiency of Banana cv Kepok<br />Amorang through Modifications of Culture Media and<br />Incubation Temperature. Yati Supriati. The budless<br />banana cv Kepok Amorang is potentially commercialized<br />due to its sweet taste and does not have flower bud, hence<br />reduced the potential of being infected by the blood disease<br />pathogen. Enhancement of banana industry needs continuous<br />supplies of large number banana seedlings. In vitro<br />culture enable the production of seedlings in a large scale,<br />uniform, quick. The research aims: (1) to formulate an<br />efficient medium for in vitro multiplication of cv Kepok<br />Amorang shoot, (2) to identify efficient growth environment<br />for in vitro culture of cv Kepok Amorang, and (3) to formulate<br />an efficient culture medium for roots inductions of cv<br />Kepok Amorang. The plant material used was in vitro culture<br />of Kepok cv Amorang, 2 cm in height without leaf and root.<br />The media formulation for shoot multiplication were full<br />strength, half strength, one fourth strength MS media,<br />supplemented with either 1, 3, or 5 ppm IBA. On optimization<br />step, the media tested were MS, Knop, Knop and<br />Heller, Hyponex N, Growmore N, and Rosasol N containing<br />of 1 ppm BA. The explants were incubated in culture room<br />with 8, 12, and 16 hours photoperiod with temperatures 30oC<br />(non air conditioned) and 25oC (air conditioned). The root<br />induction trial was done using MS, Knop, Knop and Heller,<br />Hyponex N, Growmore N, and Rosasol N media containing<br />of 1 ppm and 3 ppm IBA. The results showed that the best<br />medium formula for shoot multiplication was ¼ MS + 1 ppm<br />IBA. The best incubation condition was 16 hours photoperiods<br />at 30oC. The best media for root induction was<br />Hyponex 2 g/l + 1 ppm IBA. This culture method reduced<br />cost by Rp 261.7 per plantlet through efficiency of media<br />formulation and electricity use.</p>

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