Callus Diameter Research Articles

The use of auxin to regenerate embryogenic callus of Indonesian local garlic varieties

Abstract Garlic is a national priority commodity with regards to reducing imports. One of the obstacles in its national production is the limitation of seed quality. This study aimed to investigate how auxin impacts the regeneration of embryogenic callus in three local garlic varieties. The experiment employed a two-factor Complete Randomized Design, involving varieties (Lumbu Putih, Geol, and Sangga Sembalun) and auxin types: A (IAA 1 ppm), B (IAA 3 ppm), C (IAA 5 ppm), D (IBA 1 ppm), E (IBA 3 ppm), and F (IBA 5 ppm). Each treatment was repeated three times so that the total treatment was 54 experimental units. Embryogenic calli, weighing about 0.2 g, were cultivated on the treatment medium. The observed variables were the diameter and texture of the callus, as well as the number of green spots on the callus. The results showed no significant interaction between variety and auxin treatment, but variety had a significant effect on callus diameter. The largest callus diameter was shown by the Sangga Sembalun variety under treatment C (2.75 mm) with a friable texture, yellowish color, and rooted structure, while the smallest callus diameter was produced by the Geol variety in treatment E (1.20 mm) with a non-friable texture and white color. The highest number of green spots was also produced from Sangga Sembalun in treatment D (7 green spots). It was concluded that auxin plays a role in the regeneration of somatic embryos in garlic, but its effectiveness depends on the genotype.

Optimalisation of in vitro sterilisation methods for North Sumatra local garlic (Allium Sativum L.)

Abstract The establishment stage is critical to the success of plant tissue culture. Each plant tissue has unique tissue surface characteristics that interact with various natural microbes. The goal of this study was to find a sterilisation method that is effective at removing contamination while not damaging or inhibiting explant regeneration. In this study, Completely Randomized Design (CRD) factorial with 2 factors was used. The first factor was the disinfectant, namely T1 (Benzalkonium chloride 0.5%), T2 (Nordox 6 g/L 0.6%), and T3 (Clorox 10%). The second factor is the type of media, consisting of M1 (BDS supplemented with 1.8 μM/L 2,4-D and 9.2 μM/L Kinetin), M2 (BDS supplemented with 9 μM/L 2,4-D and 9.2 μM/L Kinetin), M3 (MS supplemented with 9 μM/L 2,4-D and 4.6 μM/L Kinetin), and M4 (MS media supplemented with 1.3 μM/L 2,4-D and 9.2 μM/L Kinetin. The observed parameters were the percentage of contamination, the survival rate, the type of contamination, the percentage of callus growing, weight and diameter. The T2 treatment was the most effective in reducing contamination and had the highest survival rate. T3 treatment resulted in the highest callus regeneration percentage, callus weight, and callus diameter, but also the highest level of contamination.

In vitro selection and characterization of cadmium-tolerant calli of Tagetes erecta and Gomphrena globosa

Ornamental plants (OPs) are beneficial to remove, control and reduce heavy metals (HMs) in a process called 'phytoremediation'. This study evaluated the in vitro system-based phytoremediation properties of Tagetes erecta and Gomphrena globosa calli. Leaves from in vitro seed-grown T. erecta and G. globosa were used as an explant source for callus culture. Callus culture was found optimal in MS medium supplemented with 8 μM BAP + NAA for T. erecta and 2 μ M 2, 4-D for G. globosa. These plants were grown in their respective optimized (controlled) medium enriched with different amounts (50, 100, 150, 200, and 250 μM) of Cadmium (Cd) added in the form of Cadmium chloride (CdCl2). The in vitro calli developed were evaluated for Cd stress tolerance based on callus diameter, growth tolerance index (GTI) and catalase (CAT) activity. Over four weeks, the callus diameters of T. erecta and G. globosa grown in different concentrations of Cd had lower growth than that of the controlled one. On the other hand, the GTI measured were greatest at 150 μM of Cd for both T. erecta (130.95%) and G. globosa (149.32%) suggesting a potential Cd tolerance. However, the CAT activity in T. erecta callus increased with the Cd concentration peaking at 150 µM then started declining while G. globosa callus showed the highest CAT activity at 50 μM of Cd. Thus, T. erecta callus showed greater Cd tolerance with the prospect of utilizing it for phytoremediation. The study also suggests growing T. erecta at 150 μM Cd for tolerant calli.

Efficacy of Randu Honey Supplementation on Callus Formation in Closed Fractures of the Tibia in Wistar Rats

Introduction: Tibial fracture is one of the most common injuries. Randu honey has the potential to speed up fracture healing. This study aims to determine the efficacy of randu honey supplementation on callus formation in closed fractures of the tibia in Wistar rats.
 Methods: This research uses a true experimental design with post test only control group design. A total of 36 Wistar rats were divided into two groups, namely the control group and the treatment group which were given 200g/BW(gram)/day of randu honey. Fractures were made on the rat tibia and callus diameter was measured at 3 weeks after treatment.
 Results: The callus diameter in the treatment group was significantly larger than the control group (p<0.05) in radiological and macroscopic measurements.
 Conclusion: Supplementation with randu honey is effective in the formation of callus in closed fractures of the tibia in Wistar rats.

Effects of Elicitation on Invitro Regeneration of two Tomato (Solanum lycopersicum L.) Cultivars in Tissue Culture

Exploring alternative avenues, in vitro culture emerges as a promising option for potential bioactive compound sources. However, compared to intact plants, only a few cultures demonstrate efficient synthesis of secondary metabolites. Elicitors have gained prominence as stress agents for enhancing in vitro micropropagation in specific tissues, organs, and cells. Recent advancements in plant tissue culture involve elicitors, opening new possibilities for in vitro production of crucial food crops. This research aimed to investigate the impact of three elicitors (Activane®, Micobiol®, and Stemicol®) on germination and in vitro multiplication of two tomato cultivars explants, employing both direct and indirect in vitro organogenesis. Among the texted elicitors, Micobiol® emerged as a successful elicitor, promoting optimal seed germination, survival, and 100% growth compared to the 80% in the control group. Further, Activane® exhibited a favourable induction response and achieved 96%, 95%, and 100% in weight and diameter of callus, yet various elicitor concentrations did not exert significant influence across treatments. In conclusion, an effective disinfection and in vitro implantation of tomato seeds ensured successful germination, promoting seedling survival and growth. Various elicitors positively impacted in vitro organogenesis, particularly in root induction, with higher survival percentages in acclimatized plants. The study guides future research on elicitor treatments for large-scale tomato in vitro propagation, emphasizing the need to identify optimal elicitor concentrations.

The Effect of NAA, BAP and NAA, BAP Combinations on the Growth of In-vitro Culturally Promoted Leaves of Sansevieria ehrenbergii

Sansevieria is an ornamental succulent leaf plant of the family Agavaceae, having a beautiful and diverse shape and color. Propagation of Sansevieria can be done by tissue culture method with the addition of cytokinins and auxins to MS media. The study used a complete randomized design consisting of 8 treatments, 3 tests, there were 24 experimental units. The observed parameters are when the callus appears, the color of the callus, the texture of the callus, the weight of the callus, the diameter of the callus, and the percentage of success in growing. Data analysis used variance analysis and tested using 5% DNMRT. The data is processed using a Statistical Analysis System (SAS). The results showed that the application of NAA, BAP, and the combination of NAA and BAP affected the per-plant explant of Sansevieria ehrenbergii leaves on all observation parameters. The treatment of NAA 2 mg.l-1 + BAP 1 mg.l-1 gave effect tending to be faster when callus appeared with an average of 75.75 HST, NAA 1 mg.l-1 + BAP 2 mg.l-1 m. and tends to be larger in callus diameter with an average of 0.9033 g and 19.133 mm and NAA 1 mg.l-1 + BAP 1 mg.l-1 gives a higher influence on the percentage of callus growth by an average of 100 %.

Gefitinib enhances healing of long bone fractures in rats via inhibition of the epidermal growth factor receptor signal pathway

Purpose: To investigate the influence of gefitinib on healing of long bone fractures in rats, and the involvement of epidermal growth factor receptor (EGFR) signal route in the process.
 Methods: A model of long bone fracture was established in 30 Sprague-Dawley (SD) rats which were allocated to control and study groups (n = 15/group). Rats in study group received gefitinib (100 mg/kg per day) via gavage, while control rats were given 0.55 % methylcellulose daily for 7 - 42 days. Fracture healing, maximum callus diameter, serum levels of bone markers and mRNA levels of bone turnover indices in the callus, were determined.
 Results: Following 1 week of therapy, fracture lines were clear in both groups, and callus was produced. On days 7, 14, 21, and 28, maximum callus diameter of study group was significantly higher than that of control group (p < 0.05). However, there was no significant difference in the maximum callus diameter between the two groups after 42 days (p > 0.05). On days 7, 14 and 21, there were significantly higher BALP and PINP levels, and TRACP-5b and CTX values in study group than in control (p < 0.05). On days 7 and 14, study group COLa1 mRNA and osteocalcin (OC) mRNA was significantly raised, relative to control values. On days 7, 14 and 21, COL10 mRNA expression was significantly up-regulated in study group relative to controls (p < 0.05). However, mRNA expression levels of COLa1, OC, COL10, and COL2A1 were similar in both groups at other time points.
 Conclusion: Gefitinib enhances the healing of long bone fractures and callus formation in rats, probably through inhibition of EGFR signaling pathway. Therefore, gefitinib is beneficial in bone formation in rats.

The effects of hydroxychloroquine-induced oxidative stress on fracture healing in an experimental rat model.

The purpose of this study was to investigate whether hydroxychloroquine (HCQ) sulfate causes oxidative stress (OS) and its effect on fracture healing in an experimental rat model. In this experimental study, open diaphyseal femur fractures were induced in 24 eight-week-old male rats (mean weight: 225±25 g; range, 200 to 250 g) and then fixed with K-wire. The rats were divided into four groups: HCQ-2, control-2 (C-2), HCQ-4, and control-4 (C-4). During the study period, rats in the HCQ groups received an HCQ solution (160 mg/kg/day), whereas rats in the control groups received saline. The HCQ-2 and C-2 groups were sacrificed on the 14th day, and the HCQ-4 and C-4 groups were sacrificed on the 28th day. After sacrifice, malondialdehyde levels induced by OS were calculated for each rat, and fracture healing was evaluated radiographically, histomorphometrically, histopathologically, and immunohistochemically. Malondialdehyde levels were higher in the HCQ groups than in the control groups (p<0.05). Hydroxychloroquine caused OS in rats. The ratio of total callus diameter to femur bone diameter was lower in HCQ groups compared to control groups (p<0.05). No differences were observed when comparing radiological and histological healing results between the control and HCQ groups. Alkaline phosphatase levels were lower in the HCQ-4 group than the C-4 group at week four (p<0.05), although osteocalcin and osteopontin levels did not differ between groups (p>0.05). Oxidative stress had no adverse effects on histologic healing outcomes and osteoblast functions. Cathepsin K and tartrate-resistant acid phosphatase-5b levels were higher in the HCQ-4 group than in the C-4 group (p<0.05). While the number and function of osteoclasts increased due to OS in callus tissue, a decrease in the number of chondrocytes was observed. Hydroxychloroquine-induced OS increases the number and function of osteoclasts and decreases the number of hypertrophic chondrocytes and endochondral ossification but has no significant effect on mid-late osteoblast products and histological fracture healing scores.

Respon Pertumbuhan Tanaman Porang (Amorphophallus muelleri Blume.) Dengan Penambahan NAA (Naphthalene Acetic Acid) dan BAP (6-Benzyl Amino Purine) Pada Perbanyakan Secara In Vitro

Porang is one of the export crops that are much hunted in several countries. The increasing number of needs has resulted in limited porang seeds so that there is a need for propagation of porang plants with good quality, one of which is by propagation through tissue culture or in vitro. In order to propagate porang plants in vitro, this research tries to discover the ideal concentration of NAA and BAP. From August to December 2022, this study was conducted in the Plant Tissue Culture Laboratory of Jember State Polytechnic. The experiment employed a parametric statistical design with 2 treatments: 0.5 mg/L NAA + 1.5 mg/L BAP (P1) and 0.5 mg/L + 2.5 mg/L BAP (P2), with each treatment being repeated 10 times. According to the findings, the parameters of callus emergence time (46 HST) and callus diameter (1.51 cm) were best achieved by administering NAA and BAP at a dosage of 0.5 mg/L NAA + 2.5 mg/L BAP. According to the study aims, the optimal concentration for explant growth and development is believed to be 0.5 mg/l NAA + 2.5 mg/l BAP.

Development of somatic embryogenesis of mangrove fan palm (Licuala spinosa Thunb.) from culturing immature zygotic embryo

AbstractMangrove fan palm is an endemic plant that grows well and has for a long period been related to traditional ceremonies in southern Thailand. It is an endangered plant with a slow growth and is difficult to propagate in mass production. Tissue culture technique can be used to produce a large number of plants within a short time and to preserve in vitro germplasm resulting in the plant not becoming extinct from natural habitats. Thus, the objectives of this research were to study effects of auxins and embryo preparation on somatic embryogenesis. The sterile immature zygotic embryo was cultured on MS medium with different types and concentrations of auxins. The results showed that 1 mg/l dicamba gave the best results in callus induction at 100% and callus diameter at 4.67 mm after 8 weeks of culture. For callus proliferation, callus chopped at 100 times and transferred to 0.1 mg/l dicamba containing oil palm culture medium gave the highest callus diameter at 10.33 mm after 4 weeks of culture and somatic embryos were also produced. Upon transferring somatic embryos to MS medium with 0.2 M sorbitol, a large number of various developmental stages of secondary somatic embryos were induced within 4 weeks. Therefore, the finding of this study could be highlighted that dicamba and chopping callus have an important role to induce callus and somatic embryo. This is the first report achieved on in vitro propagation of this plant and it will be beneficial for genetic conservation and biotechnological applications in the future.

GWAS across multiple environments and WGCNA suggest the involvement of ZmARF23 in embryonic callus induction from immature maize embryos.

Combined GWAS, WGCNA, and gene-based association studies identified the co-expression network and hub genes for maize EC induction. ZmARF23 bound to ZmSAUR15 promoter and regulated its expression, affecting EC induction. Embryonic callus (EC) induction in immature maize embryos shows high genotype dependence, which limits the application of genetic transformation in transgenic breeding and gene function elucidation in maize. Herein, we conducted a genome-wide association mapping (GWAS) for four EC induction-related traits, namely rate of embryonic callus induction (REC), increased callus diameter (ICD), ratio of shoot formation (RSF), and length of shoot (LS) across different environments. A total of 77 SNPs were significantly associated these traits under three environments and using the averages (across environments). Among these significant SNPs, five were simultaneously detected under multiple environments and 11 had respective phenotypic variation explained > 10%. A total of 257 genes were located in the linkage disequilibrium decay of these REC- and ICD-associated SNPs, of which 178 were responsive to EC induction. According to the expression values of the 178 genes, we performed a weighted gene co-expression network analysis (WGCNA) and revealed an EC induction-associated module and five hub genes. Hub gene-based association studies uncovered that the intragenic variations in GRMZM2G105473 and ZmARF23 influenced EC induction efficiency among different maize lines. Dual-luciferase reporter assay indicated that ZmARF23 bound to the promoter of a known causal gene (ZmSAUR15) for EC induction and positively regulated its expression on the transcription level. Our study will deepen the understanding of genetic and molecular mechanisms underlying EC induction and contribute to the use of genetic transformation in maize.

Radiographic, Biomechanical and Histological Characterization of Femoral Fracture Healing in Aged CD-1 Mice.

With a gradually increasing elderly population, the treatment of geriatric patients represents a major challenge for trauma and reconstructive surgery. Although, it is well established that aging affects bone metabolism, it is still controversial if aging impairs bone healing. Accordingly, we investigated fracture healing in young adult (3-4 months) and aged (16-18 months) CD-1 mice using a stable closed femoral fracture model. Bone healing was analyzed by radiographic, biomechanical and histological analysis at 1, 2, 3, 4 and 5 weeks after fracture. Our results demonstrated an increased callus diameter to femoral diameter ratio in aged animals at later time points of fracture healing when compared to young adult mice. Moreover, our biomechanical analysis revealed a significantly decreased bending stiffness at 3 and 4 weeks after fracture in aged animals. In contrast, at 5 weeks after fracture, the analysis showed no significant difference in bending stiffness between the two study groups. Additional histological analysis showed a delayed endochondral ossification in aged animals as well as a higher amounts of fibrous tissue at early healing time points. These findings indicate a delayed process of callus remodeling in aged CD-1 mice, resulting in a delayed fracture healing when compared to young adult animals. However, the overall healing capacity of the fractured femora was not affected by aging.

Propagation of Gembili (Dioscorea esculenta. L) Accession from West Papua by In-Vitro Callus Induction

Diversification of tuber foods is expected to help food security and improve public health. Gembili tuber glucomannan contains high fiber. Gembili seeds are becoming scarce, causing gembili plants to be very rarely found in the community. In the West Papua region, local people still rely on conventional supplies which are feared to threaten the gembili plant population. Tissue Culture Technology is very necessary in multiplying gembili seeds quickly and without knowing the season, by growing callus, plant propagation does not require a lot of explants to produce a large number of seeds. The research was conducted at the Politeknik Negeri Lampung Tissue Culture Laboratory, from May to July 2022. The research used a randomized block design, with each treatment repeated three times. The results showed that the composition of the media with the addition of 2.4 D 2 mL.L-1 produces an average of 8.00 days of callus growth, however, for callus growth the culture media with the addition of 2.4 D mL.L-1 produces a value best. 1.73 cm for callus diameter and a value of 0.22 g for callus weight.

Possible Effect on the Recovery of Grafted Mango Seedlings Using Naphthalene Acetic Acid

The application of naphthalene acetic acid (NAA) of different concentrations (25, 50, and 75 ppm) was applied to grafted mango seedlings to identify its effect on callus formation, callus diameter, shoot emergence, and plant survival. NAA effectively reduced the number of days to callus formation, mainly using 75 ppm. Lower than this concentration exhibited a comparable callus formation, the same as the untreated seedlings. The callus diameter, shoot emergence, and plant survival showed no significant effect by applying NAA, even with how high or low the concentration was. Low and higher than the concentrations used, young scions may be used for further investigation. Likewise, other plant growth regulators should be used to hasten the callus development and higher plant recovery rate in grafted mango seedlings.

IN VITRO PLANT REGENERATION IN LOCAL AUS RICE THROUGH MATURED SEED CULTURE

An investigation was carried out to decipher the effect of genotypes and concentrations of growth regulators for callus induction and subsequent plant regeneration from mature seeds of rice. Two local rice cultivars viz. Nayanmony and Kalomona and two growth regulators viz. 2, 4-D and NAA were used to evaluate their performance. The seeds were cultured under aseptic condition onto MS basal medium at varying concentrations and combinations of 2, 4-D (1.0, 1.5 and 2.0 mgL- 1) and NAA (0.0, 0.5 and 1.0 mgL-1) for callus induction. For plant regeneration MS medium fortified with 0.5 mgL- 1 NAA and 3.0 mgL-1 BAP was applied. Between the two cultivars Nayanmony showed better responses in callus induction frequency (91.38%), weight of callus (0.90 g)as well as diameter of callus (7.48 mm). Among the combinations 2, 4-D (1.5 mgL-1) alone produced the best callus (95.16%) and the highest weight of callus (1.88 g) but diameter of callus (8.27 mm) was the highest from 2, 4-D 1.0 mgL-1 +0.5 mgL-1 NAA. It was observed that both genotype and growth regulator significantly affected callus induction and plant regeneration. The medium supplemented with 2, 4-D 1.5 mgL-1 + NAA0.0 mgL-1 exhibited better performance in callus induction (99.33%) and weight of callus (1.92 g) in cultivar Nayanmony whereas Kalomona was better for diameter of callus (8.38 mm) at 2, 4-D 1.0 mgL-1 + NAA0.5 mgL-1. The cultivar Nayanmony performed better regarding plant regeneration (88.15%) compared to Kalomona (86.67%). Plantlets derived from the 2, 4-D 1.5 mgL-1+ NAA 0.0 mgL-1 showed the best results for survival. Therefore, it is suggested that application of MS medium supplemented with 2,4-D 1.5 mgL-1 and NAA 0.0 mgL-1 is advantageous to accomplish overall efficiency of callus induction, subsequent plant regeneration and survival from seeds of Nayanmony cultivars. Thus, selection of better responsive cultivar like Nayanmony and MS medium fortified with 1.5 to 2.0 mgL-1 2, 4-D would be beneficial in transformation attempts to improve the crop.

Combined QTL Mapping across Multiple Environments and Co-Expression Network Analysis Identified Key Genes for Embryogenic Callus Induction from Immature Maize Embryos.

The ability of immature embryos to induce embryogenic callus (EC) is crucial for genetic transformation in maize, which is highly genotype-dependent. To dissect the genetic basis of maize EC induction, we conducted QTL mapping for four EC induction-related traits, the rate of embryogenic callus induction (REC), rate of shoot formation (RSF), length of shoot (LS), and diameter of callus (DC) under three environments by using an IBM Syn10 DH population derived from a cross of B73 and Mo17. These EC induction traits showed high broad-sense heritability (>80%), and significantly negative correlations were observed between REC and each of the other traits across multiple environments. A total of 41 QTLs for EC induction were identified, among which 13, 12, 10, and 6 QTLs were responsible for DC, RSF, LS, and REC, respectively. Among them, three major QTLs accounted for >10% of the phenotypic variation, including qLS1-1 (11.54%), qLS1-3 (10.68%), and qREC4-1 (11.45%). Based on the expression data of the 215 candidate genes located in these QTL intervals, we performed a weighted gene co-expression network analysis (WGCNA). A combined use of KEGG pathway enrichment and eigengene-based connectivity (KME) values identified the EC induction-associated module and four hub genes (Zm00001d028477, Zm00001d047896, Zm00001d034388, and Zm00001d022542). Gene-based association analyses validated that the variations in Zm00001d028477 and Zm00001d034388, which were involved in tryptophan biosynthesis and metabolism, respectively, significantly affected EC induction ability among different inbred lines. Our study brings novel insights into the genetic and molecular mechanisms of EC induction and helps to promote marker-assisted selection of high-REC varieties in maize.

The Effect of Diclofenac Sodium on Callus Formation in White Male Rat (Rattus norvegicus) Cruris Fracture Healing

Highlights: Sodium diclofenac is one of NSAID a common treatment to relieve pain associated with bone fractures. Sodium diclofenac with a some dose of body weight could decrease the callus quality on fracture healing. Abstract: Non-steroidal anti-inflammatory drugs (NSAIDs), such as diclofenac sodium, are standard treatments to relieve pain associated with bone fractures. The bone healing process consists of four stages: inflammation, soft callus formation, complex callus formation, and bone remodeling. Previous studies mentioned that intake of NSAIDs (sodium diclofenac) could inhibit the bone healing process. This study examined the effect of diclofenac sodium intake on callus formation in fracture healing. In this study, thirty-six rats (Rattus Norvegicus) with fractures were used and divided into two groups, namely 18 rats for the control and 18 rats for the treatment group. In the treatment group, each rat was given 1.8 mg sodium diclofenac/150 grams of body weight per day. In the control group, each rat was given CMC-Na 0.5% with equal volume as diclofenac sodium in the treatment group. After 28 days, all the rats were stunned until dead, and the diameter and strength of their calluses were measured. In the treatment group with diclofenac sodium1.8 mg/ 150 grams BW/ 28 days after the tibia bone callus was pressed using the Shimadzu tool, the lowest callus strength was found to be 56.500 N, and the highest callus strength was 59.000 N. The lowest callus diameter in the treatment group was 4 mm, the highest was 5 mm. In the control group, the lowest callus strength was 76 N, and the highest callus strength was 77 N. The lowest callus diameter in the control group was 6 mm, and the highest was 8 mm. The strongest callus in the treatment group was found in the sixth observation, with a value of 59 N and a diameter of 4 mm. In the control group, the highest callus strength was 77 N, with a diameter of 7-8 mm. These measurements were found on the 5th, 7th, 8th, 9th, 16th, and 17th observations. Diclofenac sodium with a dose of 1.8 mg/150 grams of body weight could decrease the callus quality parameters, such as callus strength and diameter on fracture healing.

Brassinosteroid Combined With Indolbutyric Acid in Blueberry Micropropagation

The lack of availability of good quality seedlings for blueberry cultivation is an obstacle in the market, preventing the increase of production and cultivated areas. In order to improve rooting of blueberry in vitro, different concentrations of BIOBRAS 16&amp;reg; associated with indolbutyric acid in blueberry micropropagation were evaluated. For such a purpose, the Wood Plant Medium (WPM) culture medium plus the following plant regulators: 0.1, 0.3 and 0.5 mg L-1 brassinosteroids (BIOBRAS 16&amp;reg;) in conjunction with indolbutyric acid (IBA) concentrations of 1.3 and 5 mg L-1, with four replications were taken into account. At the end of 82 days of cultivation and development of the explants in these culture media, the following response variables were assessed: callus percentage (CP), callus diameter (CD), rooting percentage (RP), number of shoots (NS), number of leaves (NL), shoot length (SL), root length (RL) and fresh mass of shoots (FMS). CP was found to be stimulated in so far as concentrations of BIOBRAS 16&amp;reg; and IBA increased up to 0.5 mg L-1 and their diameter increased at concentrations of 3.0 and 5.0 mg L-1 for IBA. Thus, it is concluded that a combination of 0.3 mg L-1 BIOBRAS 16&amp;reg; combined with IBA concentrations of 3.0 and 5.0 mg L-1 contributes to promote root growth and rises in leaf number and fresh mass of micro-propagated blueberry. The use of a 0.3 mg L-1 of BIOBRAS 16&amp;reg; associated with concentrations of 3.0 and 5.0 mg L-1 of IBA showed a high percentage of root formation in blueberry.

The effect of Aloe vera ethanolic extract in preventing ischemic-reperfusion injury during long bone fracture healing after tourniquet application: An animal study

Background: Tourniquet is a common instrument used in Orthopedics field to reduce blood loss, providing a better operating field. One of the deleterious effects of tourniquet is ischemic-reperfusion injury, which occurs after deflation of the tourniquet. One of the possible ways to mitigate ischemic-reperfusion injury is by administrating antioxidants to reduce oxidative stress. Natural ingredients like Aloe vera are well known for its antioxidant and anti-inflammatory activities. This experimental study was conducted by fracturing the tibia of male Wistar strain rats (Rattus norvegicus) and administering Aloe vera gel orally as antioxidant. Method: A total of 18 rats were used in this study, divided into 6 groups. The first group was the control group, that included rats with fractured tibia not receiving torniquet and aloe vera treatment. The second and third groups were rats with fracture, who received torniquet application for one hour and two hours respectively without aloe vera treatment. The rest were the treatment groups, the rats with factures were given different dosages of Aloe vera extract: 40 mg/kg bodyweight (BW), 60 mg/kg BW, and 80 mg/kg BW. Three hours after administration, the tourniquet was inflated for two hours, followed by its deflation. The tibia was harvested to examine levels of superoxide dismutase (SOD), Malondialdehyde (MDA), and Bone morphogenetic protein 7 (BMP-7), and callus diameter, and osteoblast numbers were evaluated. Result: Administration of Aloe vera extract reduced the oxidative stress parameters (SOD and MDA). Tourniquet application decreased the osteoblast cell count and callus diameter, and administration of Aloe vera extract increased both variables close to the control value (p&lt;0.05). Conclusion: The result of this study suggests that Aloe vera extract could be used to ameliorate ischemic-reperfusion injury.

Various Concentration Distribution of Benzil Amino Purine Concentration on Callus Growth in Apple Cucumber Originated from Karawang

&#x0D; The purpose of the study is to find the best Benzil Amino Purine (BAP) concentrations and proper explants for the growth of the apple cucumber callus (Cucumis sp.) on Murashige and Skoog (MS) media. The research method used was an experimental method with nonparametric statistics with treatments A0B1 (BAP 0 + Leaves), A0B2 (BAP 0 + Stem), A1B1 (BAP 0.5 μM + Leaf), A1B2 (BAP 0.5 μM + Stem), A2B1 (BAP 1.5 μM + Leaf), A2B2 (BAP 1.5 μM + Stem), A3B1 (BAP 3.0 μM+ Leaves) and A3B2 (BAP 3.0 μM+ Stems) and analyzed descriptively using the Kruskal Wallis Test. The parameters observed were callus emergence, callus diameter, growth rate and callus morphology. The results showed accretion of an apple cucumber callus at concentration 0.5 μM BAP + stem which exerted a faster effect on callus formation time (27 hsi), 1.5 μM BAP + stem giving the largest diameter with 1.2 cm. The stem explant gives the best growth rate (0.8 cm/3 day) as well as concentrations of 0.5 μM and 1.5 μM giving the best growth rate (0.462 cm/3 day and 0.887 cm/3 day). This result showed calluses A0B2 and A1B1 (compact + green) as well as A1B2 and A2B2 (crumb + yellowish-green).