scholarly journals Temporary Immersion System Improves Regeneration of in vitro Irradiated Recalcitrant Rice Embryogenic Calli

2021 ◽  
Author(s):  
Alejandro Hernández-Soto ◽  
Jason Pérez-Chávez ◽  
Rebeca Fait-Zuñiga ◽  
Randall Rojas-Vásquez ◽  
Andres Gatica-Arias ◽  
...  
Keyword(s):  
Gamma Ray ◽  
Lethal Dose ◽  
Radiation Condition ◽  
Embryogenic Calli ◽  
Temporary Immersion System ◽  
Semisolid Medium ◽  
Regeneration Rate ◽  
In Vitro Establishment

The development of gamma rays mutant rice lines would be a solution for introducing variability in already farmer using varieties. In vitro gamma (60Co) mutagenesis reduces chimeras and allows a faster selection of desired traits but requires laboratory process optimization. The objective of the present work was the in vitro establishment of a recalcitrant rice embryogenic calli, the determination of its sensitivity to gamma radiation (Co-60), sequencing MATK and Rubisco for identification purposes, as well as generation optimization. The radiosensitivity of embryogenic calli resulted in an LD50 of 110Gy, while the 20% lethal dose was 64Gy. All sequenced genes matched perfectly with already reported MATK and Rubisco O. sativa genes with a clear SNP that identifies the local variety related to the southeast Asia Region. Callus induction improves with an MS with 2mg/L 2,4D, and the regeneration was achieved with an MS medium with 3mg/L BAP and 0,5mg/L NAA. The optimized radiation condition was 60Gy with an 83% regeneration in a semisolid medium, allowing a balance between mutation and regeneration. When increased to 80Gy, the regeneration rate falls to 29%. An immersion system (RITA®) of either 60 or 120 seconds every 8hours allowed a systematic and homogeneous total regeneration of the recalcitrant line, in contrast with the semisolid medium that resulted in positive but irregular regeneration. Other well-known recalcitrant cultivars, CR1821, CR1113 also had an improving regeneration in the immersion system, demonstrating its potential use for recalcitrant materials. To our knowledge, this is the first report on using an immersion system to allow regeneration of gamma-ray mutants from recalcitrant rice materials.

scholarly journals Determination of Median Lethal Concentrations (Lc50) of Pathogenic Antigens and Their Applications in Vaccine Development

2020 ◽  
Author(s):  
Saganuwan Alhaji Saganuwan
Keyword(s):  
Staphylococcus Aureus ◽  
Disease Virus ◽  
Vaccine Development ◽  
Hepatitis A Virus ◽  
Lethal Dose ◽  
Vaccine Dose ◽  
Foot And Mouth Disease ◽  
Mouth Disease Virus

Abstract Objective: Lack of ideal mathematical models to qualify and quantify both pathogenicity, and virulence is a dreadful setback in development of new antimicrobials and vaccines against resistance pathogenic microorganisms. Hence, the modified arithmetical formula of Reed and Muench has been integrated with other formulas and used for determination of antigen concentration and parasites inoculums that would kill 50% of test animals (LC50).Results: Microorganisms’ antigens tested are Staphylococcus aureus, Streptococcus pneumonia, Pseudomonas aeruginosa in mice and rat, Edwardsiella ictaluri, Aeromonas hydrophila, Aeromonas veronii in fish, New Castle Disease virus in chicken, Sheep Pox Virus, Foot-and-Mouth Disease Virus and Hepatitis A virus in vitro, respectively. The LC50s for the pathogens using different routes of administrations are 1.93 x 103 (sheep poxvirus) and 1.75 x 1010 for Staphylococcus aureus (ATCC29213) in rat respectively. N is the number of vaccine dose that could neutralize the LC50.Titre index (TI) equals N log10 LC50 and provides protection against lethal dose in graded fashion which translates to protection index. Hence, parasite inoculum of 103 to 1011 could be used as basis for median lethal dose (LD50), LC50 and median bacterial concentrations (BC50) determination, pathogenic dose for immune stimulation should be sought at concentrations less than LC10.

scholarly journals PENGARUH IRADIASI SINAR GAMMA [60Co] TERHADAP BACTROCERA CARAMBOLAE DREW & HANCOCK IN VITRO DAN IN VIVO

2015 ◽  
Vol 15 (1) ◽  
pp. 17
Author(s):  
Endang Sri Ratna ◽  
Kemas Usman ◽  
Indah Arastuti ◽  
Dadan Hindayana
Keyword(s):  
Gamma Ray ◽  
Lethal Dose ◽  
Fruit Fly ◽  
Probit Analysis ◽  
Adult Survival ◽  
Significance Level ◽  
Bactrocera Carambolae ◽  
Gamma Ray Irradiation

Effect of gamma irradiation [60Co] against Bactrocera carambolae Drew & Hancock in vitro and in vivo. Bactrocera carambolae Drew & Hancock is one of the most important pests on guava fruit. According to a quarantine regulation in export-import commodities, irradiation treatment is a suitable methods for eradicating infested organism, which is relatively safe for the environment. The aim of this research was to determine mortality doses and an effective dose of [60Co] gamma ray irradiation for the eradication purpose, and its implication on the survival of fruit fly B. carambolae. Two irradiation methods of in vitro dan in vivo were carried out, by exposing egg and 3rd instar larvae of B. carambolae obtained from the laboratory reared insect. Eleven doses of gamma ray irradiation of 0, 30, 50, 75, 100, 125, 150, 175, 200, 300, 450, and 600 Gy were applied, respectively. The level of 99% fruit fly mortality was estimated by the value of LD99 using probit analysis and the number of larvae, pupae and adult survival were evaluated by analysis of variance (ANOVA), and the means compared by Tukey’s test, at 5% of significance level. These result showed that the effective lethal dose (LD99) of irradiation that could be successful to eradicate eggs and 3rd instar larvae in vitro were 2225 and 2343 Gy and in vivo were 3165 dan 3177 Gy, respectively. Almost all of the treated larvae survived and developed to pupae, therefore only the minimum irradiation dose of 30 Gy allowed the pupae to develop into adults.

scholarly journals OPTIMASI DAN EVALUASI METODE KRIOPRESERVASI PURWOCENG

2020 ◽  
Vol 19 (3) ◽  
pp. 147
Author(s):  
IKA ROOSTIKA ◽  
IRENG DARWATI ◽  
RITA MEGIA
Keyword(s):  
Visual Observation ◽  
Sem Analysis ◽  
Combined Treatments ◽  
Embryogenic Calli ◽  
Regeneration Rate ◽  
Adenine Sulphate ◽  
In Vitro Shoots ◽  
Better Than

<p>ABSTRAK<br />Optimasi dan evaluasi metode kriopreservasi perlu dilakukan dalam<br />menentukan protokol standar untuk penyimpanan jangka panjang biakan<br />purwoceng. Penelitian ini bertujuan untuk mengetahui pengaruh kombinasi<br />perlakuan pratumbuh, prakultur, dan formulasi media pemulih terhadap<br />daya tumbuh dan daya regenerasi tunas in vitro dan kalus embriogenik<br />serta untuk mengevaluasi metode kriopreservasi melalui observasi<br />morfologi, anatomi, dan sitologi. Penelitian dilakukan di Laboratorium<br />Kultur Jaringan Kelompok Peneliti Biologi Sel dan Jaringan BB Litbang<br />Biogen pada tahun 2008-2009. Teknik kriopreservasi yang digunakan<br />adalah vitrifikasi (untuk apeks) dan enkapsulasi-vitrifikasi (untuk kalus<br />embriogenik). Pada teknik vitrifikasi, tunas pucuk diberi perlakuan<br />pratumbuh dengan sukrosa (3, 4, 5, dan 6%) selama 1 dan 2 minggu,<br />perlakuan prakultur dilakukan pada media yang mengandung sukrosa 0,3<br />M selama 1 dan 3 hari, perlakuan dehidrasi dengan PVS2 diberikan selama<br />15 dan 30 menit, dan media pemulih yang diujikan adalah media dasar MS<br />atau DKW dengan dan tanpa penambahan adenin sulfat 20 ppm. Pada<br />teknik enkapsulasi-vitrifikasi, kalus embriogenik dienkapsulasi terlebih<br />dahulu dengan Na-alginat 3%, perlakuan dehidrasi dengan PVS2 diberikan<br />selama 0, 30, dan 60 menit. Evaluasi metode teknik kriopreservasi<br />dilakukan melalui pengamatan morfologi secara visual, anatomi meristem<br />dengan scanning electron microscope (SEM), pengujian viabilitas dengan<br />fluorescein diacetate (FDA), dan analisis ploidi secara flowcytometry.<br />Hasil penelitian menunjukkan bahwa teknik enkapsulasi-vitrifikasi lebih<br />baik daripada teknik vitrifikasi untuk kriopreservasi purwoceng. Walaupun<br />persentase keberhasilan kriopreservasi rendah (10%), kalus embriogenik<br />purwoceng mampu berproliferasi dan beregenerasi menjadi ribuan embrio<br />somatik dewasa. Evaluasi metode kriopreservasi dengan SEM dan FDA<br />dapat diterapkan untuk memperkirakan keberhasilan teknik kriopreservasi<br />secara dini sedangkan analisis flowcytometry dapat diterapkan untuk<br />menguji stabilitas genetik bahan tanaman pasca-kriopreservasi.<br />Kata kunci: Pimpinella pruatjan Molk., kriopreservasi, SEM, FDA,<br />flowcytometry</p><p>ABSTRACT<br />Optimization and evaluation of cryopreservation methods should be<br />conducted to obtain standard protocol for long term conservation of<br />pruatjan. The objective of this study was to evaluate the effect of<br />combined treatments of pregrowth, preculture, and recovery media to the<br />survival and regeneration rate of in vitro shoots and embryogenic calli and<br />to evaluate the cryopreservation methods by observing the morphological,<br />anatomical, and cytological characters. The techniques of vitrification (for<br />apex) and encapsulation-vitrification (for embryogenic calli) were applied<br />in this study. On vitrification technique, the apical shoots were pregrown<br />on media containing of 3, 4, 5, and 6% sucrose for 1 and 2 weeks,<br />precultured on media containing of 0,3 M sucrose for 1 and 3 days,<br />dehydrated by PVS2 solution for 15 and 30 minutes, and planted on<br />recovery media (MS or DKW basal media supplemented with 20 ppm<br />adenine sulphate). On encapsulation-vitrification technique, embryogenic<br />calli were encapsulated by 3% Na-alginate, dehydrated by PVS2 solution<br />for 0, 30, and 60 minutes. The evaluation of cryopreservation methods was<br />done through visual observation, SEM analysis, viability test, and<br />flowcytometry determination. The result showed that encapsulation-<br />vitrification was better than vitrification technique for cryopreservation of<br />pruatjan. The successful rate of this method was low (10%) but the<br />embryogenic calli could proliferate and regenerate into thousands mature<br />somatic embryos. The evaluation by SEM and FDA can be applied as<br />early detection to estimate the successful of cryopreservation, whereas<br />flowcytometry  analysis  may  determine  the  genetic  stability  of<br />cryopreserved materials.<br />Key words: Pimpinella pruatjan Molk., cryopreservation, SEM, FDA,<br />flowcytometry</p>

scholarly journals Evaluation of Bioactive Compounds, in Vitro Antioxidant Activity and Acute Toxicity of Ethanol Extract of Morinda lucida Leaves

2021 ◽  
pp. 32-38
Author(s):  
F. M. Idih ◽  
O. V. Alagbe ◽  
V. D. Sheneni ◽  
J. Ebune
Keyword(s):  
Antioxidant Activity ◽  
Bioactive Compounds ◽  
Lethal Dose ◽  
Ethanol Extract ◽  
Gas Chromatography Mass Spectrometry ◽  
Medium Size ◽  
Traditional Treatment ◽  
In Vitro Antioxidant Activity

Plants are known to contain phytochemicals of pharmacological relevance and as such have been utilized in the treatment and management of various diseases. Morinda lucida, a medium size tropical tree belonging to the rubiaceae family and widely distributed in Africa is one of these plants. It has been reportedly used in the traditional treatment and management of diseases. This study is aimed at identifying compounds with pharmacological relevance in the ethanol extract of Morinda lucida leaves, the antioxidant activity and lethal dose determination of the extract. The leaves of Morinda lucida was extracted with ethanol; phytochemical and bioactive compounds analysis, in vitro antioxidant activity and lethal dose (LD50) determinations were carried out. It was observed in the study that the extract contains alkaloids, quinines, quinones, flavonoids and tannins. The gas chromatography mass spectrometry (GC-MS) identified phenol 2, 4-bis (1,1-dimethylethyl) (2.82%), Stilbenes (12.32%), Phenoxazine (2.60%) and Benz(cd) indol-2(1H)-one, 1-methyl- (2.60%) amongst other compounds in the extract. The in vitro antioxidant activity evaluation of the extract revealed that it possesses a significant antioxidant activity which increased with increasing concentration. The LD50 determination revealed the extract was safe as there was no death recorded even at a dose as high as 5000 mg/kg. This study shows that Morinda lucida possesses enormous pharmacological potentials.

Determination of the Lethal Dose of Dexamethasone for Early Passage In Vitro Human Glioblastoma Cell Cultures

Neurosurgery ◽  
1993 ◽  
Vol 33 (3) ◽  
pp. 485-488 ◽  
Author(s):  
Robert J. Maciunas ◽  
Robert A. Mericle ◽  
Christopher L. Sneed ◽  
Deborah J. Hefner ◽  
Patricia A. Commers ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Lethal Dose ◽  
Glioblastoma Cell ◽  
Early Passage ◽  
Human Glioblastoma ◽  
Human Glioblastoma Cell

scholarly journals Formation of upland rice drought-tolerant mutants by mutation induction and in vitro selection

2020 ◽  
Vol 21 (4) ◽  
Author(s):  
Rossa Yunita ◽  
ISWARI S. DEWI ◽  
ENDANG GATI LESTARI ◽  
RAGAPADMI PURNAMANENGSIH ◽  
SUCI RAHAYU ◽  
...  
Keyword(s):  
In Vitro Selection ◽  
Gamma Ray ◽  
Upland Rice ◽  
Mutation Induction ◽  
Selection Methods ◽  
Rice Varieties ◽  
Drought Tolerant ◽  
Gamma Ray Irradiation

Abstract. Yunita R, Dewi IS, Lestari EG, Purnamanengsih R, Rahayu S, Mastur. 2020. Formation of upland rice drought-tolerant mutants by mutation induction and in vitro selection. Biodiversitas 21: 1476-1482. Increased production can be done by increasing the planting area, but the available land is sub-optimal land with drought stress. Drought-tolerant rice varieties are needed to utilize this land. To create drought-tolerant rice can use induction mutation and in vitro selection methods. This study aims to obtain tolerant upland rice mutants through mutation and in vitro selection. The parents used were Batutegi and Situpatenggang varieties. This research consisted of several main activities, namely callus induction, determination of LD 50% value ofPEG determination of LC 50% value of gamma-ray irradiation, mutation induction and in vitro selection, bud regeneration and acclimatization. The results of this study are mutant callus formed by gamma-ray irradiation at a dose of 24.68 Gy for Situpatenggang and 22.15 Gy for Batutegi (LD50) and selected on PEG media at a dose of 24.11% for Situpatenggang and 25.18% for Batutegi (LC50). The mutant callus regenerated on MS + BA 3 mg/L + Zeatin 0.1 mg/L produced 83 Situpatenggang shoots and 73 Batutegi shoots and successfully acclimatized were 52 Situpatenggang lines and 49 Batuteg linesBatutegi

scholarly journals Development of an optimum proliferation medium via the graph kernel statistical analysis method for genetically stable in vitro propagation of endemic Thymus cilicicus (Turkey)

2021 ◽  
Vol 80 (2) ◽  
Author(s):  
Ergun Kaya ◽  
Selin Galatali ◽  
Irem Aktay ◽  
Onur Celik ◽  
Bilge Ozturk ◽  
...  
Keyword(s):  
In Vitro Propagation ◽  
Kernel Method ◽  
Eastern Mediterranean ◽  
Medium Composition ◽  
Regeneration Rate ◽  
Graph Kernel ◽  
Proliferation Medium ◽  
Shoot Number

Thymus cilicicus is an endemic Eastern Mediterranean element that has aromatic-medicinal properties. Its natural population spreads across gravelly ground and open rocky areas of South and Southwest Anatolia. The current study on in vitro propagation of T. cilicicus focused deeply on environmental applications such as the development of an optimum medium composition for efficient and genetically stable micropropagation and improved preservation procedures for long-time conservation of elite germplasms for further studies. For this purpose, MS and OM media were used individually and in combination with cytokinins, charcoal, AgNO3, Fe-EDDHA, and H3BO3. The raw data were statistically analyzed via the graph kernel method to optimize the nonlinear relationship between all parameters. The optimal proliferation medium for T. cilicicus was OM supplemented with a combination of 10 g L-1 charcoal and 1 mg L-1 KIN and the calculated averages of the best regeneration rate, the best shoot number and the best shoot length were 96.89%, 3 and 1.24 respectively on this medium. The determination of genetic stability of in vitro grown plants on the optimum medium compositions obtained by the graph kernel method was carried out with the use of the ISSR-PCR technique. All the ISSR primers produced a total of 192 reproductive band profiles, none of which were polymorphic. Furthermore, the micropropagated plants were successfully rooted and acclimatized to greenhouse conditions. In this study, we present a graph kernel multiple propagation index which considers all the possible parameters needing to be analyzed. Such an index is used for the first time for the determination of the optimum proliferation medium.

scholarly journals Effects of gamma rays from 60Co source on dwarf Dendrobium cross-breeding

2020 ◽  
Vol 19 (01) ◽  
pp. 32-40
Author(s):  
Vinh V. Nguyen
Keyword(s):  
Gamma Rays ◽  
Gamma Ray ◽  
Lethal Dose ◽  
Survival Ratio ◽  
Cross Breeding ◽  
60Co Source ◽  
60Co Gamma ◽  
Effective Doses ◽  
Higher Doses

In this research, seeds of DM12x13 dwarf Dendrobium cross-breeding germinated on (1 )/2 MS medium after 40 days were irradiated by 60Co source with various doses to determine LD50 (Lethal Dose 50%). After LD50 had been determined, protocorms of DM12x13 dwarf Dendrobium cross-breeding was irradiated by gamma rays from 60Co source in 5 doses: 0 Gy (control), 20 Gy, 40 Gy, 60 Gy and 80 Gy, dose rate 90 Gy/h to study the effect of gamma radiation on mutagenicity and evaluate the growth of these mutations in vitro. The result showed that the lethal dose of hybrid DM12x13 Dendrobium protocorm under the effect of 60Co gamma ray was 68 Gy. Survival ratio, growth and the ability to growth of plants reduced in higher doses and dead ratio were 100% when irradiated more than 80 Gy after 7 months. The effective doses for treating protocorm of DM12x13 crossbred lines were 20, 40 and 60 Gy. These doses were suitable for increased frequency of variation with generating wide-spread, diverse in structure and color of stems and leaves

Determination of the Lethal Dose of Dexamethasone for Early Passage In Vitro Human Glioblastoma Cell Cultures

Neurosurgery ◽  
1993 ◽  
Vol 33 (3) ◽  
pp. 485-488 ◽  
Author(s):  
Robert J. Maciunas ◽  
Robert A. Mericle ◽  
Christopher L. Sneed ◽  
Deborah J. Hefner ◽  
Patricia A. Commers ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Lethal Dose ◽  
Glioblastoma Cell ◽  
Early Passage ◽  
Human Glioblastoma ◽  
Human Glioblastoma Cell

scholarly journals Effects of human serum albumin on post-mortem changes of malathion

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yoshikazu Yamagishi ◽  
Hirotaro Iwase ◽  
Yasumitsu Ogra
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Lethal Dose ◽  
Free Form ◽  
Post Mortem ◽  
Flight Mass Spectrometry ◽  
Dose Dependent Fashion

AbstractMalathion, diethyl 2-[(dimethoxyphosphinothioyl)thio]butanedioate, is one of most widely used organophosphoryl pesticide, and it has been detected in several clinical cases of accidental exposure and suicide. It is reported that the observed malathion concentration in blood of persons who suffer from malathion poisoning is smaller than the expected concentration. Because malathion is bound to human serum albumin (HSA), recovery of malathion in the free form is insufficient. We detected malathion adducts in HSA by liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q/TOF–MS). The mass spectra showed that malathion was preferably bound to the lysine (K) and cysteinylproline (CP) residues of HSA. The K- and CP-adducts of malathion were increased in vitro with a dose-dependent fashion when its concentration was smaller than the lethal dose. Further, the K-adduct was also detected in post-mortem blood of an autopsied subject suffering from intentional malathion ingestion. These results suggest that the K-adduct seems to be available to use a biomarker of malathion poisoning, and the determination of the K-adduct could make possible to estimate the amount of malathion ingestion.

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