Analysis of expression of m 6A regulators in colorectal tumor samples based on TCGA and GEO databases / 中国基层医药

Objective:To detect the expression of N6-methyladenosine (m 6A) regulators in colorectal tumor samples and its clinical significance. Methods:From September to December 2021, the data regarding the expression of m 6A regulators in colorectal tumor samples were collected using The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) and the expression of m 6A regulators was compared between different clinical pathological characteristics and between different molecular subtypes. Survival analysis was conducted in patients with colorectal cancer with different m 6A expression levels. Results:In TCGA, insulin-like growth factor 2 mRNA binding protein 1 (IGF2BP1), insulin-like growth factor 2 mRNA binding protein 3 (IGF2BP3), and YTH N6-methyladenosine RNA binding protein 1 (YTHDF1) were higher in colorectal tumor samples than normal tissue samples (TCGA-COAD: IGF2BP3: 12.80 vs. 204.46, logFC = 4.00, P = 0.003; YTHDF1: 2 347.56 vs. 3712.77, logFC = 0.66, P < 0.001; TCGA-READ: IGF2BP1: 6.20 vs. 359.32, logFC = 5.82, P = 0.007; YTHDF1: 2 470.10 vs. 4 369.09, logFC = 0.82, P = 0.020). The intersection of TCGA and GEO databases showed that methyltransferase like 14 (METTL14), YTH domain m 6A RNA binding protein 3 (YTHDF3), and α-ketoglutarate dependent dioxygenase AlkB homolog 5 (ALKBH5) were downregulated in colorectal tumor samples compared with normal tissue samples (TCGA-COAD: METTL14: 1 051.56 vs. 711.40, logFC = -0.56, P < 0.001; YTHDF3:4 613.85 vs. 3 155.05, logFC = -0.55, P < 0.001, ALKBH5: 4 250.10 vs. 2 555.55, logFC = -0.73, P < 0.001; TCGA-READ vs. METTL14: 1 113.3 vs. 674.36, logFC = -0.72, P < 0.001; YTHDF3: 5 034.30 vs. 3 331.95, logFC = -0.60, P = 0.004; ALKBH5: 4 902.20 vs. 2 529.71, logFC = -0.95, P < 0.001; GEO-CRC vs. METTL14: 6.58 vs. 6.33, logFC = -0.06, P < 0.001; YTHDF3: 6.28 vs. 6.20, logFC = -0.02, P = 0.002; ALKBH5: 5.07 vs. 4.98, logFC = -0.02, P < 0.001). In colorectal tumor samples of different molecular subtypes, IGF2BP2, HNRNPC, TP53 and YTHDF2 expression was low in KRAS (IGF2BP2: 48.53 vs. 44.04, t = 2.64, P = 0.008; HNRNPC: 121.30 vs. 112.60, t = 2.32, P = 0.020; TP53: 65.30 vs. 60.26, t = 2.11, P = 0.034; YTHDF2: 54.07 vs. 51.19; t = 1.97, P = 0.048). Different clinical pathological characteristics showed that IGF2BP1 was higher in colorectal cancer with positive lymph nodes (8.97 vs. 6.11, W = 20 008, P = 0.002), distant metastasis (8.94 vs. 6.41, W = 19 104, P = 0.009), or stage Ⅲ-Ⅳ (7.46 vs. 7.13, W = 8 779, P = 0.025) than normal tissue. ALKBH5 overexpression, advanced age, presence of vascular invasion, and late pathological staging were significantly related to a short survival period (high ALKBH5 expression vs. low ALKBH5 expression: 5.85 years vs. not available, HR: 1.63, 95% CI: 1.071-2.491, P = 0.021; > 68 years vs. < 68 years: 6.78 years vs. not available, HR: 1.59, 95% CI: 1.049-2.422, P = 0.047; stage Ⅲ-Ⅳ vs. stage Ⅰ-Ⅱ: 4.55 years vs. 8.33 years, HR: 2.89, 95% CI: 1.895-4.425, P < 0.001; presence of vein invasion vs. absence of vein invasion: 5.48 years vs. not available, HR: 2.82, 95% CI: 1.672-4.783, P < 0.001). Conclusion:Some of the m 6A regulators are associated with the biological characteristics and prognosis of colorectal cancer.

IN VITRO GERMINATION AND VEGETATIVE PROPAGATION THROUGH BUD DEVELOPMENT OF SACHA INCHI (Plukenetia volubilis) / Germinación y propagación vegetativa in vitro a través del desarrollo de yemas de Sacha Inchi (Plukenetia volubilis)

ABSTRACT This study describes the in vitro seed germination and micropropagation of Plukenetia volubilis (sacha inchi), an oilseed crop rich in omega-3 fatty acids, with health benefits and several industrial applications. Seed germination was evaluated in different culture media (MS and 1/2 MS), seed coat presence/absence and culture temperature (18 °C and 28 °C). Micropropagation was performed using axillary bud development (ABD) on nodal segments from in vitro seedlings. KIN, BAP and 2-ip were evaluated for ABD, and the effect of modified MS in 453 mg L-1 CaCl2 and 351.62 mg L-1 MgSO4 on ABD and shoot survival was assessed to improve the process. Finally, six treatments were evaluated to optimize ABD and shoot leaf formation. Seed germination of 91.6 % was achieved in MS at 28 °C when the seed coat was removed. ABD was obtained in 45 % and 40 % with 0.4 mg L-1 KIN and 0.6 mg L-1 2-ip, respectively, with the least CAL. The modification in 453 mg L-1 CaCl2 then allowed 76 % ABD and 82 % explant survival. ABD response was optimized to 95 % and 2.45 leaves with MS medium + CaCl2 modification + 10 % coconut water + 0.4 mg L-1 KIN. The same results were obtained by replacing the latter with 0.6 mg L-1 2-ip. Rooting was achieved in MS without PGR, and acclimatization was successful. The results indicate that plant production via germination and vegetative propagation is effective for commercial purposes.

RESUMEN Este estudio describe la germinación in vitro y micropropagación de Plukenetia volubilis, un cultivo oleaginoso rico en omega-3 benéfico para la salud y con múltiples aplicaciones industriales. Se evaluó en la germinación diferentes medios de cultivo (MS y 1/2 MS), presencia-ausencia de testa y temperatura de cultivo (18 ° C y 28 ° C). La micropropagación se realizó vía yemas axilares (ABD) de plántulas in vitro. Se evaluó el efecto de KIN, BAP y 2-ip sobre ABD, seguidamente, para mejorar el proceso se evaluó el efecto de MS modificado en 453 mg L-1 CaCl2 y 351.62 mg L-1 MgSO4 sobre ABD y supervivencia del brote. Finalmente, se evaluaron seis tratamientos para optimizar ABD y la formación de hojas. Se logró una germinación 91,6 % en MS a 28 °C cuando se retiró la testa. Se obtuvo 45 % y 40 % de ABD con 0,4 mg L-1 KIN y 0,6 mg L-1 2-ip respectivamente, ambos con la menor CAL. Posteriormente, la modificación de CaCl2 permitió 76 % ABD y 82 % de supervivencia. Se optimizó ABD al 95 % con 2,45 hojas por brote con el medio: MS + modificación de CaCl2 + 10 % de agua de coco + 0.4 mg L-1 KIN, los mismos resultados se obtuvieron cambiando este último con 0,6 mg L-1 2-ip, se logró enraizamiento en MS sin PGR, y la aclimatización fue exitosa. Los resultados indican que la producción de plantas vía germinación y propagación vegetativa es efectiva con fines comerciales.

Effect of Rhustox on Micropropagation of Scoparia dulcis L. through Leaf Explants Culture

Aim: To investigate the effect of Rhus toxicodendron (30CH) along with different compositions of phytohormones (Auxin and Cytokinin) on the basis of growth and multiplication of explants under optimum temperature under in-vitro conditions. Study Design: To establish and design the standard protocol for the in-vitro propagation through leaf explant of Scoparia dulcis under stress of phytohormones and homeopathic medicine Rhus toxicodendron (30CH). Place and Duration of Study: The plant materials were procured from the Herbal Botanical Garden Patna Science College, Department of Botany, Patna University, Patna, Bihar. The experimental part was carried out in Plant Tissue Culture Laboratory, between December 2017 to August 2018 in Department of Botany P.U. Patna. Methodlogy: The sterilized leaf explants were inoculated into MS media fortified with different phytohormones (Auxin and Cytokinin) and Rhus tox(30CH) under aseptic environmental conditions for the growth and development of callus, embryoids etc. Result: The explants in MS medium supplemented with auxins phytohormones and Rhus tox(30CH) exhibited that IAA (0.10 to 2.0 mg/l) and BAP (0.10 to 2.5 mg/l) induces green and compact calli. Whereas at 0.30mg/l of IAA and 0.50 mg/l BAP induced brown friable calli. 2,4-D (1.5 mg/l) and Kinetin (1.5-6.5mg/l) concentrations induced brown and friable calli. Rhus tox(30CH) (100 µl/100 ml) enhances proliferation with 2,4-D and Kinetin (1.5/1.5 mg/l.). Conclusion: After 42 days of culture initiation and establishment the callus was 520.0±1.12 mg in the mixture of 2,4-D and Kinetin (1.5 mg/l) in Rhus tox free medium. Whereas weight of callus were found to be 1092±0.74 mg after 42 days in the same medium of 2, 4-D and Kinetin (1.5/5.5 mg/l) supplemented with Rhus tox (100 µl/100 ml). Hence, the investigation proponded that the Rhus tox (CH30) has increased the rate of callus development and plantlet regeneration.

Kernel size and weight affected by three plant bioregulators applied at bloom to Non Pareil and Carmel almond cultivars

Background: The yield of almonds [Prunus dulcis (Mill.) D.A. Webb] could be low due to climatic problems and any factor improving kernel size and weight, such as the use of plant bioregulators (PBRs), should be beneficial. Results: Three plant bioregulators: 24-epibrassinolide (BL), gibberellic acid (GA3) and kinetin (KN) were applied at three spray concentrations to Non Pareil and Carmel cultivars, at two phenological stages during bloom, in the 2014 and 2015 seasons. The results showed significant differences (P b 0.0001). For total dry weight of Non Pareil, the best treatment was BL (30 mg·L-1), with an average of 1.45 g, while the control was 1.30 g, at pink button during 2015. For Carmel, the best dry weight was 1.23 g, achieved with BL (30 mg·L-1) at fallen petals in both seasons. The average dry weight of the controls varied between 1.13 and 1.18 g. The greatest almond lengths and widths in Non Pareil were 24.98 mm and 15.05 mm, achieved with BL (30 mg·L-1) and KN (50 µL·L-1) treatments, respectively, applied at pink button in 2015. In Carmel, the greatest length and width were 24.38 and 13.44 mm, obtained with BL (30 mg·L-1) applied at the stages of pink button and fallen petals, respectively, in 2015. The control reached lengths between 22.33 and 23.38 mm, and widths between 11.99 and 12.93 mm. Conclusions: The use of the bioregulators showed significant favorable effects on dry weight, length and width of kernels at harvest, in both cultivars.

Kinetin Improves Barrier Function of the Skin by Modulating Keratinocyte Differentiation Markers

BACKGROUND: Kinetin is a plant hormone that regulates growth and differentiation. Keratinocytes, the basic building blocks of the epidermis, function in maintaining the skin barrier. OBJECTIVE: We examined whether kinetin induces skin barrier functions in vitro and in vivo. METHODS: To evaluate the efficacy of kinetin at the cellular level, expression of keratinocyte differentiation markers was assessed. Moreover, we examined the clinical efficacy of kinetin by evaluating skin moisture, transepidermal water loss (TEWL), and skin surface roughness in patients who used kinetin-containing cream. We performed quantitative real-time polymerase chain reaction to measure the expression of keratinocyte differentiation markers in HaCaT cells following treatment. A clinical trial was performed to assess skin moisture, TEWL, and evenness of skin texture in subjects who used kinetin-containing cream for 4 weeks. RESULTS: Kinetin increased involucrin, and keratin 1 mRNA in HaCaT cells. Moreover, use of a kinetin-containing cream improved skin moisture and TEWL while decreasing roughness of skin texture. CONCLUSION: Kinetin induced the expression of keratinocyte differentiation markers, suggesting that it may affect differentiation to improve skin moisture content, TEWL, and other signs of skin aging. Therefore, kinetin is a potential new component for use in cosmetics as an anti-aging agent that improves the barrier function of skin.

Effects of variations in culture media and hormonal treatments upon callus induction potential in endosperm explant of Barringtonia racemosa L

Objective: To induce callus from the medicinally valuable species, Barringtonia racemosa L. (B. racemosa) whereby the formation of callus is essential for micropropagation studies and in vitro plant secondary metabolites production. Methods: The callus induction potential in B. racemosa was assessed from endosperm explant cultured on different culture media and plant hormonal treatments. Lloyd and McCown's woody plant medium and Murashige and Skoog's medium were used in the study as culture media. On the other hand, various concentrations and combinations of 2,4-dichlorophenoxyacetic acid (1.0-2.0 mg/L) and kinetin (0.5-2.5 mg/L) had been incorporated in the culture media to exert the effects of auxin and cytokinin on callus induction. Results: From the present study, it was found that the profuse [(1.681 ± 0.770) g fresh weight, (0.239 ± 0.239) g dry weight] and friable callus formation was optimally produced with desirable morphology and considerable percentage of callus induction (56.70%) in endosperm explants cultured on 1.0 mg/L 2,4-dichlorophenoxyacetic acid and 1.5 mg/L kinetin in Murashige and Skoog's medium. Conclusions: A reliable protocol for inducing callus formation of profuse and friable morphology in endosperm explant of B. racemosa had therefore been successfully established.

A microdroplet cell culture based high frequency somatic embryogenesis system for pigeonpea, Cajanus cajan (L.) Millsp.

A protocol for high frequency production of somatic embryos was worked out in pigeonpea, Cajanus cajan (L.) Millsp. The protocol involved sequential employment of embryogenic callus cultures, low density cell suspension cultures and a novel microdroplet cell culture system. The microdroplet cell cultures involved culture of a single cell in 10 µl of Murashige and Skoog’s medium supplemented with phytohormones, growth factors and phospholipid precursors. By employing the microdroplet cell cultures, single cells in isolation were grown into cell clones which developed somatic embryos. Further, 2,4-dichlorophenoxyacetic acid, kinetin, polyethylene glycol, putrescine, spermine, spermidine, choline chloride, ethanolamine and LiCl were supplemented to the low density cell suspension cultures and microdroplet cell cultures to screen for their cell division and somatic embryogenesis activity. Incubation of callus or the inoculum employed for low density cell suspension cultures and microdroplet cell cultures with polyethylene glycol was found critical for induction of somatic embryogenesis. Somatic embryogenesis at a frequency of 1.19, 3.16 and 6.51 per 106 cells was achieved in the callus, low density cell suspension cultures and microdroplet cell cultures, respectively. Advantages of employing microdroplet cell cultures for high frequency production of somatic embryos and its application in genetic transformation protocols are discussed.

Efficacy and safety of 0.1% kinetin cream in the treatment of photoaging skin.

Background: Kinetin is a plant‑derived compound, which is reported to possess antiaging properties. It has been used in a topical cream to manage facial photo‑damage and aging. Although studies elsewhere have shown its benefits, not many studies of the effects of kinetin in Asian skin are available. Objectives: To assess the efficacy and tolerability of 0.1% kinetin cream in the treatment of facial photo‑aging. Methods: The study was designed to be open‑label and single‑blinded, without a control group. One hundred Thai female and male subjects with mild, moderate or severe facial photo‑damage were enrolled. They were asked to apply 0.1% kinetin cream twice daily for 12 weeks and follow up at 4, 8, and 12 weeks. Subjective patient self‑assessment and physician assessment of facial skin photo‑damage were accompanied by digital photographic analysis using the VISIA® (Canfield Scientific Inc, Fairfield, NJ) imaging system. Results: At baseline, most patients reported moderate skin changes related to photo‑damage, skin texture, skin color and wrinkles. After 12 weeks, physician and patient assessments showed slight but statistically significant improvements in overall skin condition, skin texture, color, and wrinkles. Findings were similar with the digital photographic system analysis, especially in relation to skin color. Facial ultraviolet spots and redness also showed statistically significant improvements after 12 weeks. The treatment was generally well tolerated. Limitations: The study was designed to be pragmatic and hence no randomization was carried out; there were also no intrapatient or interpatient control observations, and no comparison arm. Conclusion: Kinetin (0.1%) cream was found to slightly improve cutaneous facial photo‑damage after 12 weeks of use in a group of Thai patients.

In Vitro Propagation of Chamomilla Recutita From Capitulum Inflorescence: A Medicinal Plant With Multiple Therapeutic Applications

A protocol has been developed for induction of somatic embryogenesis from whole inflorescence explants of Chamomilla recutita L. (chamomile). Chamomile is a well-known medicinal plant from the Asteraceae family often referred to as the “star among medicinal species.” Nowadays, it is a highly favoured medicinal plant in folk and traditional medicine. Its multitherapeutic, cosmetic and nutritional values have been established through the years of traditional and scientific use and research. Chamomile has an established domestic (Indian) and international market, which is increasing day by day. Among the various major constituents, α-bisabolol and chamazulene have been reported to be more useful than others. Chamazulene occurs in the capitula of the flowers in minute quantities and has been demonstrated to exert antiinflammatory activity in-vivo. Moreover, chamomile is a seasonal 4-5 months winter crop in India but is extensively required in various medicinal applications. Therefore, to increase the overall yield of this plant, its in-vitro propagation is needed. In the present study, somatic embryos were developed from capitulum explants after 2-4 weeks of culture on MS medium supplemented with 26.8 µM NAA and 11.5 µM Kin. The somatic embryos were further subcultured in-vitro, where new plantlets regenerated from embryos. It is concluded that in-vitro propagation is possible in case of chamomile and can be used to increase the overall yield of chamazulene present in the capitula of flowers as well as augment the overall yield of this important plant, which is conventionally propagated by seeds.

Direct Multiple Shoot Induction and Plantlet Regeneration from Cotyledonary Explants of Sapindus emarginatus Vahl (Soapnut).

germination of S. emarginatus in vitro cotyledon explants in BAP/Kn/TDZ (1.0-3.0 mg/L) supplemented MS medium and (2) in plant treatment with BAP/Kn/TDZ (3.0 mg/ L) in combination of 1AA (0.5 mg/L) of the cotyledon explants of plants and maintained under sterile conditions. While the former method resulted in as many as (7.5±8.6 shoot buds) from the cotyledonary explants within four weeks, the latter yielded on average approximately 8 shoot buds from each treated node in eight weeks. The cytokinin treatment in plant consisted of placing sterile filter paper moistened with sterile distilled water over the node and adding different concentrations of 6- benzylaminopurine. The best results for shoot bud regeneration were obtained with cotyledons, when cultured in the presence of (0.5 mg/L) IAA in combination with (3.0 mg/L). The shoots elongated and rooted directly in vermiculite after a pulse treatment with IBA (2.5 mg/L) for 15 min. Fungus growth, a serious problem in S. emarginatus tissue culture, was controlled using a fungicide, Bavistin, along with elimination of organic nutrients from the growth medium.

Kinetin inhibits proliferation of hepatic stellate cells by interrupting cell cycle and induces apoptosis by down-regulating ratio of Bcl-2/Bax / 华中科技大学学报（医学）（英德文版）

Liver fibrosis is an important health problem that can further progress into cirrhosis or liver cancer, and result in significant morbidity and mortality. Inhibiting proliferation and inducing apoptosis of hepatic stellate cells (HSCs) may be the key point to reverse liver fibrosis. At present, anti-fibrosis drugs are rare. Kinetin is a type of plant-derived cytokinin which has been reported to control differentiation and induce apoptosis of human cells. In this study, the HSCs were incubated with different concentrations of kinetin. The proliferation of rat HSCs was measured by MTT assay, cell cycle and apoptosis were analyzed by flow cytometry, and the apoptosis was examined by TUNEL method. The expression of Bcl-2 and Bax proteins was detected by immunocytochemistry staining. It was found that kinetin could markedly inhibit proliferation of HSCs. In a concentration range of 2 to 8 μg/mL, the inhibitory effects of kinetin on proliferation of HSCs were increased with the increased concentration and the extension of time (P < 0.01). Flow cytometry indicated that kinetin could inhibit the DNA synthesis from G0/G1 to S phase in a dose-dependent manner (P < 0.01). The apoptosis rates of the HSCs treated with 8, 4 and 2 μg/mL kinetin (25.62% ± 2.21%, 15.31% ± 1.9% and 6.18% ± 1.23%, respectively) were increased significantly compared with the control group (3.81% ± 0.93%) (P < 0.01). All the DNA frequency histogram in kinetin-treated groups showed obvious hypodiploid peak (sub-G1 peak), and with the increase of kinetin concentrations, the apoptosis rate of HSCs also showed a trend of increase. It was also found that kinetin could down-regulate the expression of Bcl-2, and up-regulate the expression of Bax, leading to the decreased ratio of Bcl-2/Bax significantly. The kinetin-induced apoptosis of HSCs was positively correlated with the expression of Bax, and negatively with the expression of Bcl-2. It was concluded that kinetin can inhibit activation and proliferation of HSCs by interrupting the cell cycle at G1/S restriction point and inducing apoptosis of HSCs via reducing the ratio of Bcl-2/Bax.

Development of Simple, Cost Effective Protocol for Micropropagation of Tylophora indica (Burm f.) Merill., an Important Medicinal Plant.

Aims: The present studies were initiated to develop a cost effective protocol for micropropagation, as a mean for conservation of medicinal plant- Tylophora indica (Burm f.) Merill. The plant is threatened and needs immediate conservation, therefore, the study was undertaken with following objective:  In vitro multiplication of Tylophora indica using nodal axillary bud proliferation and through organogenesis of callus. Study Design: For all experiments ten replicates were used per treatment and all the experiments were repeated three times. Data have been presented as Mean ± Standard deviation. Place and Duration of Study: Department of Plant Cell and Molecular Biology, Indian Institute of Advanced Research (IIAR), Gandhinagar, Gujarat, India. Methodology: For in vitro plant regeneration, micropropagation and organogenesis techniques were used. For micropropagation, surface sterilized nodal explants were inoculated on different shoot inducing media and further multiplication was obtained. Root containing shoots were transferred to pot containing pre autoclaved mixture of soil and soilrite. For organogenesis, surface sterilized leaf explants were inoculated on different types of callus inducing media. Results: All the nodal explants were sprouted at a very high frequency, i.e. 98% and sprouted buds elongated up to 8cm on three different media. Dissected explants grew further and average height of shoots reached 9.5cm±0.80cm within 30 days. Interestingly, root formation was observed on the same media; so that the best media was 0.4mg L-1 BA and 0.1mg L-1 Kn for both initiation as well as for multiplication. For organogenesis, the fragile callus was observed on media containing 2mg L-1 2,4-D and 0.1mg L-1 Kn. Green pigmented calli were transferred to MS media, where it regenerated in to shoots and roots, simultaneously Conclusion: The protocol of micropropagation through axillary bud proliferation described here is very simple, repetitive and cost effective, which can be easily utilized for commercial cultivation. On shoot multiplication media, root formation observed, thereby making the process is one step; which is very easy to follow.

In-Vitro Micropropagation and Media Optimization of Stevia Reboudiana an Important Bio-Sweetener and Medicinal Plant”.

The technique of Plant Biotechnology has an important role to play in the production of agriculture, horticulture and ornamental plants and in the manipulation of plants for improved agronomic performance. In the present study initiation of callus has been done by using explants from auxiliary and apical meristems of Stevia rebaudiana. Explants were inoculated on MS basal medium having Vitamin supplement with Auxin like 2-4, D and NAA (0.5mg/lt. -2.0mg/lt.) alone or in combination with cytokinis like BAP & Kinetin (0.52 mg/lt. -1.0 mg/lt.). Sucrose 30 gm. and agar 4.5 gm./lt. After the formation of explants, they were transferred into shoot induction medium containing different concentration of cytokine like BAP & Kinetin (0.5 mg/lt. to 30 mg/lt.) with additional vitamins. After few days numbers of multiple shoots were formed. Strong and elongated shoots were treated with root initiating medium i.e MS containing Auxin like IBA, NAA & activated charcoal. Maximum 60-70% callusing in 10 to 15 days was initiated on 2-4D (1-2mg/lt.) and 40-50% callusing were reported in medium containing (1.0 NAA & 0.5 mg/lt. BAP) in 25-30 days. Nodular compact cells were formed in 2-4D. The medium containing BAP alone showed shoot formation 70-80% with 40% coconut water. Second rooting medium used was containing NAA and activated charcoal and 85% rooting were observed within 7-10 days after transferring. Finally the cultured plants were transferred for hardening.

Kinetin inhibits apoptosis of aging spleen cells induced by D-galactose in rats

Kinetin (Kn) is a cytokinin growth factor that exerts several anti-aging and antioxidant effects on cells and organs. To investigate the mechanism underlying apoptotic events in aging cells induced by D-galactose (D-gal), we examined the effect of Kn delivered via nuchal subcutaneous injection on D-gal-induced aging and apoptosis in rats. Our results showed that interleukin (IL)-2 levels and mitochondrial membrane potential (DeltaPsim) were decreased by Kn in aging rats while IL-6 production and apoptosis increased. In addition, the expression of anti-apoptotic Bcl-2 was low while that of Bax was high in the aging group. After treated with Kn, compared with aging group, there showed obvious difference in Kn group with elevated IL-2, proliferation index, Bcl-2, DeltaPsim and decreased IL-6 and Bax in splenic lymphocyte. Based on these results, we concluded that Kn can effectively protect the rat spleen from aging, apoptosis, and atrophy.

Effect of cytokinins on in vitro multiplication of Sophora tonkinensis

Objective:To determine the effects of different cytokinins at various concentrations on in vitro shoot multiplication of an important medicinal plant. Methods: Nodal explants (1.5-2.0 cm) of Sophora tonkinensis were used. Multiple shoots were induced from nodal explants cultured on the Murashige and Skoog (MS) medium supplemented with 0.0, 0.5, 1.0, 2.0, 4.0, 8.0, or 16.0 μmol 2-isopentyladenine (2iP), N6 benzyladenine, kinetin or thiadiazuron. Results: Among the four investigated cytokinins, 2iP showed the best response for shoot multiplication. Maximum shoot induction (75%) was achieved on the MS medium supplemented with 2.0 μmol 2iP, with a mean number of 5.0 shoots per explant. In comparison to other cytokinins tried, 2iP showed the highest shoot elongation with a mean shoot length of 4.8 cm. Root initiation was observed within 15 d within the transfer of shoots onto the MS basal medium, and the rooting percentage was 100%with a mean number of 5.4 roots per shoot and root length of 6.2 cm over a period of 4 weeks. The healthy plants, hardened and transferred to a greenhouse for proper acclimatization, exhibited 100%survival. Conclusions:It can be summarized that 2iP is the optimal plant growth regulator for Sophora multiplication.

Efecto de dos citoquininas, ácido ascórbico y sacarosa en la obtención de plantas in vitro de Sorghum bicolor para la formación de callos / Effect of two cytokinin, ascorbic acid and sucrose to obtain in vitro shoots of sorghum for calli formation

El trabajo se realizó con el objetivo de obtener brotes in vitro vía organogénesis directa en sorgo variedad CIAP 2E-95, para la formación de callos con estructuras embriogénicas. Se tomaron semillas maduras de plantas que crecían en condiciones controladas, las que se desinfectaron y fueron colocadas a germinación en un medio de cultivo con las sales MS, mioinositol 100 mg L-1, sacarosa 3%, pH 5,7, Fitagel 2,5 g L-1. Para la multiplicación de los brotes, se estudiaron diferentes concentraciones de 6 Bencilaminopurina (6-BAP) y kinetina. El empleo en el medio de cultivo de multiplicación con las sales MS y 6- BAP 0,22 mg L-1, incrementó el número de brotes por explante y se observó la presencia de oxidación fenólica. La oxidación fenólica se contrarrestó con la adición al medio de cultivo de 50 mg L-1 de ácido ascórbico y se incrementó significativamente el número de brotes, altura y número de hojas. En la obtención de brotes in vitro de calidad para la formación de callos, estos fueron engrosados con las sales MS, sin reguladores de crecimiento, ácido ascórbico 50 mg L-1, Fitagel 2,5 g L-1, pH 5,7 y 40 g L-1 de sacarosa. La formación de callos con estructuras embriogénicas a partir del cilindro central de las hojas enrolladas del segmento más próximo a la base de los brotes engrosados se alcanzó a los 45 días de cultivo. Los segmentos del cilindro central de las hojas enrolladas constituyen una fuente segura y promisoria de explantes para la formación de callos embriogénicos.

The current work aimed to obtain in vitro shoots by direct organogenesis from sorghum variety CIAP 2E-95, for the formation of calluses with embryogenic structures. Mature seeds were collected from plants grown under controlled conditions, which were disinfected and were placed in a germination medium with MS salts, myo-inositol 100 mg L-1, 3% sucrose, pH 5.7, phytagel 2.5 g L-1. Different concentrations of 6-benzylaminopurine (6-BAP) and kinetin were studied for shoot multiplication. Using 6 - BAP 0.22 mg L-1 in the multiplication culture medium increased the number of shoots per explant, showing phenolic oxidation. Adding 50 mg L-1 of ascorbic acid to the culture medium resisted phenolic oxidation and the number of sprouts (5.0), height and number of leaves increased significantly. Quality in vitro buds for callus formation were obtained stimulating the thickening by using MS salts, without growth regulators, ascorbic acid 50 mg L-1, phytagel 2.5 g L-1 and pH 5.7 and 40 g L-1 of sucrose. The largest diameter of shoots, height, number of leaves and roots of in vitro plants, was achieved after 21 days of cultured in MS salts with 40 g L-1 sucrose. Calluses formation with embryogenic structures from the central cylinder of curled leaves of the nearest segment to the base of thickened shoots was achieved after 45 days of culture. Segments from the central cylinder of the curled leaves are a safe source for the formation of embryogenic calli.

An efficient in vitro plant regeneration of Dipteracanthus prostratus (Poir.) Nees.- a medicinal herb

Objective: This is the first attempt for an efficient plant regeneration protocol through in vitro direct organogenesis for a valuable medicinal plant, Dipteracanthus prostratus using nodal segment. Methods: Multiple shoots were induced from nodal explants cultured on Murashige and Skoog medium supplemented with kinetin (KIN), 6-benzylaminopurine (BAP) and 1-Naphthaleneacetic acid (NAA). Results: Maximum shoot responses (80%) were obtained with kinetin at 1.0 mg-1. The rate of shoot multiplication was maintained in subsequent subculture on similar fresh culture medium. The highest shoot length (3.96cm) was obtained with seventy three percentages of shoots at 0.2 mg-1 NAA along with 1.0 mg-1 kinetin. Maximum length of root (3.63cm) was formed at 0.5 mg-1 IBA with significant responses (80%). Rooted plantlets were then transferred to perforated plastic cups and grown in the green house at 80% survival rate. Conclusions: The highest survival rate was noticed and this plant developmental protocol could be used for large- scale regeneration of D. prostratus.

Establishment of an efficient callus induction and plant regeneration system in Pakistani wheat (Triticum aestivum) cultivars

Four commercially grown wheat varieties of Pakistan, namely Inqilab-91, Chakwal-97, Tatara and Manthar were used for this investigation. For callus induction different concentrations of 2,4-Dichlorophenoxyaceticacid (2,4-D) along with 0.1 mg/L of Kinetin were evaluated. For regeneration initially different concentrations of Indole-3-Acetic Acid (IAA) and 6-BenzylAminoPurine (BAP) were tested. Best hormone combinations were further subjected to Kinetin and 6-ã-ã-dimethylallylaminopurine (2iP). For Inqilab-91, Chakwal-97 and Manthar, 3 mg/L of 2,4-D was found optimum, which induced 83.25 percent, 77.75 percent and 95.20 percent of embryogenic calli, respectively. Maximum callus induction (97.18 percent) was observed in Tatara when 2 mg/L of 2,4-D was used. As regard to regeneration, Inqilab-91, Chakwal-97 and Manthar showed maximum regeneration on media containing 0.1 mg/L IAA, 0.4 mg/L Kinetin and 0.5 mg/L 2iP, regenerating 87.25 percent, 81.75 percent and 68.75 percent respectively. For Tatara maximum regeneration of 12.25 percent was obtained on 0.1 mg/L IAA and 2 mg/L of BAP. Presently optimized regeneration method holds promise for facilitating the deployment of agronomical important trait through genetic transformation for the improvement of this important food crop.

Efeitos de citocininas sobre a anatomia foliar e o crescimento de Annona glabra L. durante o cultivo in vitro e ex vitro / Effects of cytokinins on the leaf anatomy and growth of Annona glabra L. during in vitro and ex vitro culture

O efeito de diferentes fontes de citocininas durante o cultivo in vitro de A. glabra sobre características anatômicas de folhas e crescimento das plantas foi avaliado neste trabalho. BAP (6-benzilaminopurina) e KIN (cinetina) induziram aumento na espessura do mesofilo, enquanto que ZEA (zeatina) promoveu aumento na densidade e no índice estomático e no desenvolvimento do sistema vascular de folhas. A utilização de KIN e BAP proporcionou maior desenvolvimento e taxa de sobrevivência das plantas durante as fases de enraizamento e aclimatização.

The effect of different sources of cytokinins during the in vitro cultivation of A. glabra on anatomical characteristics of leaves and plant growth was evaluated in this work. BAP (6-benzilaminopurine) and KIN (kinetin) induced an increase in leaf mesophyll thickness, while the ZEA (zeatin) promoted an increase in density and stomatic index and development of leaves vascular system. The utilization of KIN and BAP improved higher plant development and survival rate during the acclimatization and rooting phases.

Otimização de protocolo para micropropagação da figueira "Roxo de Valinhos" / Protocol otimization for micropropagation of 'Roxo de Valinhos' fig tree

Foram avaliados os efeitos de alterações na concentração de sacarose, do meio WPM - Wood Plant Medium - e da variação no número de gemas por segmento com diferentes doses de cinetina. Os experimentos foram conduzidos no Laboratório de Cultura de Tecidos da Universidade Federal de Lavras, utilizando plântulas previamente estabelecidas in vitro, seguindo o delineamento experimental inteiramente casualizado, em esquema fatorial 5 x 4. Os resultados mostraram que, na multiplicação de brotações de figueira "Roxo de Valinhos", podem-se usar 100 por cento do meio WPM com adição de 10gL-1 de sacarose. Foram obtidos brotos alongados quando foi usado o meio WPM sem adição de cinetina e segmentos com uma ou duas gemas. A adição de cinetina a 0,5mgL-1 e a utilização de segmentos com três gemas promoveu maior número de brotações.

Effects of different concentration of sucrose on Wood Plant Medium -WPM and variations on number of bud/plantlets with different doses of kinetin were evaluated on this research. The experiments were carried out in the Tissue Culture Laboratory at Federal University of Lavras, using plantlets which was already established in vitro. The experimental design adopted was the complete randomized in factorial scheme 5 x 4. The results revealed that better sprouts multiplication of ‘Roxo de Valinhos' fig plants occurred in WPM medium, 100 percent salts, added with 10gL-1 of sucrose and the sprout elongation in WPM medium without kinetin taking explants within one or two buds. Adding 0.5mgL-1 of kinetin and taking explants with tree buds promoted larger sprouts number.