Browsing by Subject "In vitro"

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Open Access
Artifcial intelligence–based approaches to evaluate and optimize phytoremediation potential of in vitro regenerated aquatic macrophyte Ceratophyllum demersum L.
(2023-01-06) Aasim, M.; Ali, Seyid Amjad; Aydin, S.; Bakhsh, A.; Sogukpinar, C.; Karatas, M.; Khawar, K.M.; Aydin, M.E.
Water bodies or aquatic ecosystem are susceptible to heavy metal accumulation and can adversely afect the environment and human health especially in underdeveloped nations. Phytoremediation techniques of water bodies using aquatic plants or macrophytes are well established and are recognized as eco-friendly world over. Phytoremediation of heavy metals and other pollutants in aquatic environments can be achieved by using Ceratophyllum demersum L. — a well-known foating macrophyte. In vitro regenerated plants of C. demersum (7.5 g/L) were exposed to 24, 72, and 120 h to 0, 0.5, 1.0, 2.0, and 4.0 mg/L of cadmium (CdSO4·8H2O) in water. Results revealed signifcantly diferent relationship in terms of Cd in water, Cd uptake by plants, bioconcentration factor (BCF), and Cd removal (%) from water. The study showed that Cd uptake by plants and BCF values increased signifcantly with exposure time. The highest BCF value (3776.50) was recorded for plant samples exposed to 2 mg/L Cd for 72 h. Application of all Cd concentrations and various exposure duration yielded Cd removal (%) between the ranges of 93.8 and 98.7%. These results were predicted through artifcial intelligence–based models, namely, random forest (RF), extreme gradient boosting (XGBoost), and multilayer perceptron (MLP). The tested models predicted the results accurately, and the attained results were further validated via three diferent performance metrics. The optimal regression coefcient (R2) for the models was recorded as 0.7970 (Cd water, mg/L), 0.9661 (Cd plants, mg/kg), 0.9797 bioconcentration factor (BCF), and 0.9996 (Cd removal, %), respectively. These achieved results suggest that in vitro regenerated C. demersum can be efcaciously used for phytoremediation of Cd-contaminated aquatic environments. Likewise, the proposed modeling of phytoremediation studies can further be employed more comprehensively in future studies aimed at data prediction and optimization.
Open Access
Artificial neural network and decision tree facilitated prediction and validation of cytokinin‑auxin induced in vitro organogenesis of sorghum (Sorghum bicolor L.)
(Springer Dordrecht, 2023-04-05) Aasim, M.; Ali, Seyid Amjad; Altaf, M. T.; Ali, A.; Nadeem, M. A.; Baloch, F. S.
In this study, in vitro regeneration protocol of sorghum (Sorghum bicolor) was successfully established by using direct organogenesis from a mature zygotic embryo explant. The used basal medium encompassed Murashige and Skoog medium (MS) supplemented with 2–4 mg/L Benzylaminopurine (BAP) alone or with 0.25 mg/L Indole butyric acid (IBA) or Naphthalene acetic acid (NAA). Results demonstrated a significant impact of cytokinin-auxin on shoot count (1.24–3.46) and shoot length (2.80–3.47 cm). Maximum shoot count (3.46) and shoot length (3.97 cm) were achieved on the MS medium enriched with 2 mg/L BAP + 0.25 mg/L NAA and 2.0 mg/L BAP, respectively. To ascertain the impact of BAP alone, BAP + IBA, and BAP + NAA, the data were also analyzed by using a factorial regression model. Pareto chart and normal plots were used to check either the positive or negative impact of input variables on output variables. To further explore the association between BAP + IBA and BAP + NAA on shoot count and shoot length, contour and surface plots were also built. Three different artificial intelligence-based models along with four different performance metrics were utilized to validate the predicted results. Multilayer perceptron (MLP) model performed more efficiently (R2 = 0.799 for shoot count and R2 = 0.831 for shoot length) as compared to the decision tree-based algorithms of random forest (RF) – (R2 = 0.779 for shoot count and R2 = 0.786 for shoot length) and extreme gradient boost (XGBoost) – (R2 = 0.768 for shoot count and R2 = 0.781 for shoot length). As plant tissue culture protocol is a powerful tool for genetic engineering and genome editing of crops, integration of different artificial intelligence-based models can lead to improvement of sorghum with the aid of biotechnological tools.
Open Access
Timing of induction of cardiomyocyte differentiation for in vitro cultured mesenchymal stem cells: a perspective for emergencies
(NRC Research Press, 2009) Tokçaer-Keskin, Zeynep; Akar, A. R.; Ayaloğlu-Bütün, Fatma; Terzioğlu-Kara, Ece; Durdu, S.; Özyurda, U.; Uğur, M.; Akçalı, Kamil C.
Mesenchymal stem cells (MSCs) have the capacity to differentiate into osteoblasts, chondrocytes, adipocytes, myocytes, and cardiomyocytes. Several established methods are presently available for in vitro isolation of MSCs from bone marrow. However, the duration necessary to culture them can be a major handicap to cell-based therapies needed for such urgent cardiovascular conditions as acute myocardial infarction and acute hindlimb ischemia. The best timing of car- diomyocyte differentiation induction after MCS isolation and expansion is still an unresolved issue. Our goal was to investigate the possibility of obtaining functional cardiomyocytes from rat MSC within a shorter time period. We examined MSCs' colony-forming capacity, CD90 and CD34 immunoreactivity during the 14 days of culturing. Cardiomyocyte differentiation was induced by 5-azacytidine. Immunohistochemic staining, together with intracellular Ca2+ measurement experiments, revealed that MSCs do not differentiate into any specific cell lineage but show the characteristics of MSCs on both the 9th and 14th days of the culture. To check the potential for differentiation into cardiomyocytes, experiments with caffeine application and depolarization with KCl were performed. The cells possessed some of the specific biochemical features of contracting cells, with slightly higher capacities on the 14th day. Cells from 9th and 14th days of the culture that were treated with 5-azacytidine had a higher expression of cardiac-specific markers such as troponin I, α-sarcomeric actin, and MEF2D compared with the control groups. This study illustrates that it is possible to get functional cardiomyocytes from in vitro MSC culture in a shorter time period than previously achieved. This reduction in time may provide emergency cases with access to cell-based therapies that may have previously been unavailable.