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The role of passage numbers of donor cells in the development of Arabian Oryx – Cow interspecific somatic cell nuclear transfer embryos

  • Aiman A. Ammari EMAIL logo , Muath G. ALghadi , Ahmad R. ALhimaidi and Ramzi A. Amran
Published/Copyright: April 20, 2022
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Open Chemistry
From the journal Open Chemistry Volume 20 Issue 1

Abstract

The cloning between different animals known as interspecific somatic cell nuclear transfer (iSCNT) was carried out for endangered species. The iSCNT has been characterized by a poor success rate due to several factors that influence the formation of the SCNT in various cytoplasms. The cell cycle of the transferred somatic cell, the passage number of the cultured somatic cell, the mitochondria oocytes, and their capabilities are among these factors. This study investigates the role of the passage number of the Arabian Oryx somatic cell culture when transplanted to an enucleated domestic cow oocyte and embryo development in vitro. The fibroblast somatic cell of the Arabian Oryx was cultured for several passage lanes (3–13). The optimal passage cell number was found to be 10–13 Oryx cell lines that progressed to various cell stages up to the blastula stage. There was some variation between the different passage numbers of the oryx cell line. The 3–9 cell line did not show a good developmental stage. These could be attributed to several factors that control the iSCNT as stated by several investigators. More investigation is needed to clarify the role of factors that affect the success rate for the iSCNT.

Keywords: iSCNT; enucleation; passage; embryo; Oryx

1 Introduction

Enucleated, metaphase II sheep oocytes fused with 8- or 16-cell embryonic blastomeres were employed in the first report of embryonic nuclear transfer in mammals [1,2]. Following that, other attempts to clone cattle and other animals were made. Numerous examples of effective nuclear transfer utilizing donor cells have been published for sheep [3,4,5], cows [6,7], rabbits [8,9], pigs [10], mice [11], monkeys [12], goats [13,14], and camel [15,16]. At the morula and blastocyst stages, donor cells were also revealed to be totipotent and reprogrammable by cytoplasmic factors in cow [6] and rabbits [8]. American researchers successfully injected the somatic cells of threatened animals (Bos gaurus) into an enucleated oocyte, obtained embryo at blastula stage and transferred to a recipient’s cow [17]. In addition, Saikhun et al. [18] also succeeded to inject buffalo somatic cells into an enucleated oocyte from a species cow (Bos indicus) [19]. Furthermore, Ikumi et al. [20] succeeded in transferring a somatic cell from a whale into an in vitro cloned embryo.

The endangered grey wolf was successfully cloned from somatic cells obtained from the dead carcass [21]. In wild goats, somatic cells were transferred into the oocyte of local goats [22]. Although the use of intra- or interspecies somatic cell nuclear transfer (SCNT) has resulted in the generation of cloned offspring in a variety of mammalian species, the efficiency of somatic cell cloning remains often very or extremely low. Therefore, extensive investigations are needed to identify the determinants of SCNT-mediated cloning, which can be responsible for ameliorating the efficiency of this modern-assisted reproductive technology. Among these determinants, the most important role is played by: (1) the source of nuclear donor somatic cells (NDSCs) [23,24,25,26], (2) the abilities of nuclear recipient oocytes to be artificially activated [26,27,28], and (3) molecular interactions between mitochondrial and nuclear DNA molecules in SCNT-derived embryos [29,30,31,32]. It is also worth noting about the remarkable role of: (4) the abilities of donor cell nuclear genes to epigenomically reprogram their transcriptional activities in somatic cell-cloned embryos [33,34,35] and (5) the molecular quality and survival rates of the in vitro cultured NDSCs and SCNT-derived embryos [36,37,38,39]. Taking the above-indicated determinants affecting the efficiency of somatic cell cloning, the research is required to be undertaken to improve the epigenetic reprogrammability (i.e., dedifferentiability) of donor cell nuclei in interspecies SCNT-derived embryos that have been reconstructed with somatic cells stemming from endangered or extinct mammalian species. The technique of transferring somatic cells from threatened wild species of mammals such as the Arabian Oryx is essential in their preservation. Also, the cytoplasm of cow oocytes supports the development of embryos resulting from SCNT of different types of animals [40]. Cow and Arabian Oryx are both members of the family Bovidae. The cloning of Arabian Oryx via the injection of the enucleated oocyte from cow somatic cells is realistic.

The Arabian Oryx is the smallest member of the Oryx genus and is found across the Arabian Peninsula. They vanished from the wild in the early 1970s, but zoos and private custodians preserved them. The Arabian Oryx was categorized as endangered by the International Union for Conservation of Nature in 1986 [41]. In addition, it was the first mammal to be reclassified as vulnerable after being declared extinct in the wild in 2011 [42]. This study aims to produce cloned Arabian Oryx embryos by transferring different passages of fibroblast somatic cells from Arabian Oryx into enucleated cow oocytes.

2 Materials and methods

2.1 Chemicals

Both chemicals, media and hormones, were obtained from Sigma-Aldrich Corp. (St. Louis, MO, USA), unless otherwise specified.

2.2 Ovaries, Oocyte, and maturation

Ovaries were obtained from a local slaughterhouse in Riyadh and transported in 0.9% Sodium Chloride to the laboratory during 1–2 h. The oocytes were extracted from the ovarian follicles using a 19-gauge needle connected to syringe containing 0.5 mL of tissue culture medium (Caisson Lab. Inc., Smithfield, USA). Cumulus oocyte complexes (n 706) with more than three layers of cumulus cells and homogenous cytoplasm (Figure 1a) were matured in TCM-199 at 39°C and 5% CO2 in atmospheric air at high humidity for 24 h as mentioned in our previous research [43].

Figure 1 (a) Immature oocyte, (b) mature oocyte, (c) oocyte with FPB after 24 h (the arrow = 1st polar body) and oocyte cutting, (d) the nucleus was extracted and stained, and then examined under fluorescence microscopy, (e) donor cells from Arabian Oryx, (f) injection somatic cell, (g) 2 cell stage, (h) = 4 cell stage, (i) = 8 cell stage, (j) = 16 cell stage, (k) morula stage, and (l) blastocyst stage.
Figure 1

(a) Immature oocyte, (b) mature oocyte, (c) oocyte with FPB after 24 h (the arrow = 1st polar body) and oocyte cutting, (d) the nucleus was extracted and stained, and then examined under fluorescence microscopy, (e) donor cells from Arabian Oryx, (f) injection somatic cell, (g) 2 cell stage, (h) = 4 cell stage, (i) = 8 cell stage, (j) = 16 cell stage, (k) morula stage, and (l) blastocyst stage.

2.3 Zona dissection with micropipettes

Cumulus-enclosed oocytes were placed on a 35 mm Petri plate with hyaluronidase enzyme (300 IU/1 mL) after in vitro maturation period. Cumulus cells were manually removed from oocyte by using a glass Pasteur pipette. Then, for zona dissection, only the oocytes with an extruded first polar body (FPB) would be utilized. The directing of the oocyte was controlled by the cutting pipette. When the polar body is at 12 o’clock, an oocyte position with the holding pipette, a slot or slit in the zona pellucida (ZP) was made with the cutting pipette on the opposite side of oocyte (Figure 1c) [4].

2.4 Oocyte enucleation

Cutting pipettes were used to softly spin the oocyte to extract a little part of cytoplasm contained in the plasma membrane immediately beneath the FPB of oocytes outside the ZP. Enucleation was ascertained by separating the resultant from the oocyte and cultured primarily in TCM-199. Earle’s salt is mixed with 10% fetal bovine serum and 20 μg/2 mL Hoechst for 30 m (Sigma B2261). The ooplasm was then subjected to UV light to check for the existence of a metaphase II stage [4].

2.5 Preparation of fibroblast donor somatic cells from Arabian Oryx

Oryx ears were utilized to acquire fibroblast donor cells, which have a normal diploid karyotype, and were used to transduce cell nuclei by lanes 3–13. The fibroblast cells were cultured in Dulbecco’s modified minimal basic 0.5 μg/mL Penicillin streptomycin sigma and 10% fetal bovine serum at 39°C and 5% (CO2) in atmospheric air at high humidity. The cells were grown to 80% confluence and NT cells were separated by 0.25% trypsin-EDTA solution (Gibco 25200) (Figure 1e) [4].

2.6 Analysis of cell cycle by flow cytometry

Flow cytometry was used to determine the cell cycle of Oryx cells. Cells were extracted by centrifugation, washed once with PBS after being treated with three passages, and incubated for 6–8 h. The cells were then fixed in chilly 70% alcohol for 30 m. After that, the cell suspensions were incubated with RNase for 30 min in the dark. By using a Beckman Coulter flow cytometer, 10,000 events of PI-stained cells were recorded in the FL4 Log channel using a 675 nm band-pass filter (Cytomics FC 500, Beckman Coulter, Fullerton, CA, USA). CXP capture and analysis software was used to compute the cell cycle stages and analyze the data (Figure 2).

Figure 2 The different passage cells to make sure from their presence in the G0 stage.
Figure 2

The different passage cells to make sure from their presence in the G0 stage.

2.7 Nuclear transfer, fusion, and activation

One somatic cell from an oryx fibroblast was inserted by polar body dissection pipette into the perivitelline space of every enucleated cows’ oocytes. Immediately following NT at room temperature, the cell fusion was done via a single alternating current pulse of 0.2 kV/cm for one second followed by a single DC pulse of 2.5 kV/cm for 50 s. After fusing, 5 μM ionomycin was employed to activate selected fused SCNT at room temperature for 5 min. Then, they were cultured in SOF medium supplemented with 2 mg/500 μL BSA, 20 μL/2 mL MEM (50×), 20 μL/2 mL MEM (100×), 50 μg/mL Penicillin streptomycin, 5 μg/500 μL Cycloheximide using a 35 mm Petri dish coated by mineral oil that has been tried on embryos, for 6 h at 39°C with 5% CO2. All steps of this protocol follow the method described in a previous study (Figure 1f) [4].

2.8 In vitro culture of iSCNT embryos

After 6 h of activation, iSCNT embryos were cultured for 42 h in SOF medium supplemented with 2 mg/500 L BSA., 20 μL/2 ml MEM (50×), 20 μL/2 mL MEM (100×) 50, μg/2 mL Penicillin streptomycin using Petri dishes 35 mm coated by mineral oil at 39°C for 6 h with mixture gas 90, 5, 5% of N2, CO2, O2, respectively (Figure 1g–l) [4,43].

2.9 Statistical analysis

All results were recorded and analyzed using the Mine Tab INSTAT program. The Blastocyst and cleavage rates were analyzed by using the Chi-square test.

3 Results

In this study, the collected oocytes (n 706) injected with different passage lanes 3–13 of donor cells showed different developmental rates of iSCNT Oryx – Cow embryos. From passage 3 up to 9, was no considerable rate of development, which illustrates the collected oocyte numbers, with enucleation rate, fusion rate, embryo cleavage rate, and blastula development rate. The rate of the different passage 10 to 13 was 80, 91.2, 66.3, and 84%, respectively. The fusion rate of the different passage 10 to 13 was 43, 61.4, 71, and 70%, respectively (Table 1).

Table 1

The role of passage numbers of donor cells in the development of Arabian Oryx – Cow iSCNT embryos

Passage number No. of oocytes Enucleation rate% Fusion rate% Cleavage rate% Blastocyst rate%
Passage 10 190 125 (80) 35 (43) (8) 14% (5) 62%*
Passage 11 140 114 (91.2) 70 (61.4) (25) 57%* (0) 0%
Passage 12 271 112 (66.3) 80 (71.4) (8) 16% (1) 12%
Passage 13 105 142 (84) 100 (70.4) (17) 48%* (0) 0%

*Significant difference at p < 0.01 compared to the others without star at the same type of passage or columns.

In the passage from 10 to 13, the cleavage rate of the iSCNT embryos injection with the different passage was 14, 57, 16, and 48%, respectively. The cleavage rate of the iSCNT in passages 11 and 13 indicated a considerable increase (p < 0.01) than passage lines 10 and 12 (Table 1).

While passage line 10 produced a better blastula rate compared to the other passage lines, it was significantly higher (p < 0.01) than passage lines 11, 12, and 13 (Table 1). In general, the total embryos at the blastocyst stage were five embryos (62%) from passage 10 and one embryo (12%) from passage 12, while no embryos developed up to the blastocyst stage from passages 11 and 13 (Table 1).

4 Discussion

The failure of cloning has been related to several variables, one of which is a greater rate of oocyte degeneration. This might be owing to the kind of somatic cell employed since most of the donor cells were from skin or cumulus cells, which are not in the G0 stage, or the electrical current used to fuse the somatic cell with the oocytes, which could damage the ova’s cytoplasm. In addition, due to the long term of the fibroblast cell, the rate of development of the SCNT may be affected by in vitro culture [45].

This work demonstrated the ability of the cytoplasm of domestic cows to reprogram adult Arabian Oryx fibroblast nuclei. Interspecies somatic cell nuclear transfer embryos have the potential to develop to the blastocyst. The development rate of iSCNT rebuilt Arabian Oryx–Cow embryos can be obtained at a low rate as per Li et al. [45]. SCNT efficiency was substantially lower when embryonic germ (EG) cells with lower passage numbers (3–5) were employed as donor cells than when fetal fibroblast donor cells were used. Higher passage EG cells (numbers 9–12) produced similar blastocyst rates, implying that higher passage EG cells are better suited for SCNT. This agrees with our result regarding the use of passes more than 10. The SCNT development was dramatically reduced when lower passage (numbers 3–5) porcine EG cells were used as donor cells, but higher passage (numbers 9–12) EG cells produced a greater blastula rate. However, these results disagree with Jin et al. [44] which they got better-cloned embryo development from the first passage cells compared to 4–6 passages.

There are several factors reported about early and late passages [45,46,47], which affect the proliferation of donor somatic cell culture, such as the abnormalities in chromosomes, DNA damage, and DNA methylation. The bulk of iSCNT research has focused on establishing whether an oocyte can reprogram a foreign species’ nucleus and support the initial cleavage divisions. Fewer studies, on the other side, have looked at the variables that cause embryonic failure in less taxonomically various species during the preimplantation stages [48]. Two of the key studied challenges in iSCNT include failure to reprogram the somatic cell and incompatibility between the nucleus and the cytoplasm [49,50]. Incompatibilities between the nucleus and the cytoplasm have been found to cause harmful cellular processes as an outcome of species variation in studies on and nuclear hybrid embryos. Larger species’ divergence lengths, according to some theories, may hinder communication between the nucleus and cytoplasm components such as the mitochondrial genome [51,52].

Many studies have shown the molecular that may cause the failure of the development of the embryos it produces by iSCNT [53,54,55]. In our research, we studied two species known to produce related species (Bovidae species) through interspecies cloning, which indicates some similarities between the two species.

Furthermore, the timing of DNA replication appears to vary among SCNT embryos [56]. In addition, the cytoplasm of donated oocytes for iSCNT is from different species which have different regulations for DNA demethylation of somatic cells [57,58].

The findings of this study revealed information about some of the factors that contribute to the development of Arabian Oryx – Cow iSCNT embryos in certain passages and another passage failure in others. The reduced rate of cleavage and blastocyst formation in iSCNT might be attributed to mitochondrial function, which causes embryo development failure during the early stages of division in embryos [53,59]. The results of our study showed the best cleavage and blastula rate obtained from passages 10 and 12. There may be other reasons that are recommended for more investigation such as gene expression of the Cow–Oryx iSCNT.

5 Conclusion

Our current investigations have proved the occurrence of intergroup variability in the in vitro developmental outcomes of interspecies (oryx → bovine) cloned embryos generated using somatic cell lines at different passage numbers (between 10th and 13th passage). In contrast, the use of somatic cell lines between passages 3 and 9 has resulted in remarkable alleviations in the in vitro developmental capabilities of oryx → bovine iSCNT-derived embryos to reach the blastocyst stage. The intergroup differences between the extracorporeal developmental competencies of oryx → bovine cloned embryos might be evoked by a variety of factors that affected the iSCNT efficiency. Among them, one of the most important factors seems to be molecular cooperation between nuclear and mitochondrial genomes in interspecies cloned embryos [32,39,60]. The successful developmental rates of such embryos appear to be arising from epigenomic reprogrammability of foreign (xenogeneic) somatic cell nuclear genomes in iSCNT-derived embryos [61,62,63,64,65,66,67]. For those reasons, additional intensive studies are needed to clarify how the high impact of the abovementioned molecular factors is exerted on the ex vivo development of oryx → bovine iSCNT-derived embryos.

Acknowledgment

The Project for funding this work number (RSP-2021/232) at King Saud University, Riyadh, Saudi Arabia.

  1. Funding information: The authors sincerely acknowledge the Researchers Supporting Project for funding this work number (RSP-2021/232) at King Saud University, Riyadh, Saudi Arabia.

  2. Author contributions: Conceptualization, A.A.A. and M.G.A.; methodology, A.A.A. and R.A.A; validation, A.R.A., A.A.A.; formal analysis, A.A.A.; investigation, A.A.A. and M.G.A.; data curation, A.R.A.; writing – original draft preparation, A.A.A. and A.R.A.; writing – review and editing, A.A.A., A.R.A.; visualization, A.R.A., A.A.A., and M.Q.A.; supervision, A.R.A., and M.Q.A. All authors have read and agreed to the published version of the manuscript.

  3. Conflict of interest: The authors declare that they have no conflict of interest.

  4. Ethical approval: All of the experiments were conducted according to the Guidelines for the Institutional Animal Care and Use Committee of the Zoology Department, College of Science at King Saud University.

  5. Data availability statement: The authors confirm that the data supporting the findings of this study are available within the article.

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Received: 2022-01-28
Revised: 2022-03-28
Accepted: 2022-03-31
Published Online: 2022-04-20

© 2022 Aiman A. Ammari et al., published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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https://doi.org/10.1515/chem-2022-0153
Keywords for this article
iSCNT; enucleation; passage; embryo; Oryx
Creative Commons
BY 4.0

Articles in the same Issue

  1. Regular Articles
  2. Photocatalytic degradation of Rhodamine B in aqueous phase by bimetallic metal-organic framework M/Fe-MOF (M = Co, Cu, and Mg)
  3. Assessment of using electronic portal imaging device for analysing bolus material utilised in radiation therapy
  4. A detailed investigation on highly dense CuZr bulk metallic glasses for shielding purposes
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  45. Texturing of nanocoatings for surface acoustic wave-based sensors for volatile organic compounds
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  55. Mechanical and gamma-ray shielding examinations of Bi2O3–PbO–CdO–B2O3 glass system
  56. Machine learning-based forecasting of potability of drinking water through adaptive boosting model
  57. The potential effect of the Rumex vesicarius water seeds extract treatment on mice before and during pregnancy on the serum enzymes and the histology of kidney and liver
  58. Impact of benzimidazole functional groups on the n-doping properties of benzimidazole derivatives
  59. Extraction of red pigment from Chinese jujube peel and the antioxidant activity of the pigment extracts
  60. Flexural strength and thermal properties of carbon black nanoparticle reinforced epoxy composites obtained from waste tires
  61. A focusing study on radioprotective and antioxidant effects of Annona muricata leaf extract in the circulation and liver tissue: Clinical and experimental studies
  62. Clinical comprehensive and experimental assessment of the radioprotective effect of Annona muricata leaf extract to prevent cellular damage in the ileum tissue
  63. Effect of WC content on ultrasonic properties, thermal and electrical conductivity of WC–Co–Ni–Cr composites
  64. Influence of various class cleaning agents for prosthesis on Co–Cr alloy surface
  65. The synthesis of nanocellulose-based nanocomposites for the effective removal of hexavalent chromium ions from aqueous solution
  66. Study on the influence of physical interlayers on the remaining oil production under different development modes
  67. Optimized linear regression control of DC motor under various disturbances
  68. Influence of different sample preparation strategies on hypothesis-driven shotgun proteomic analysis of human saliva
  69. Determination of flow distance of the fluid metal due to fluidity in ductile iron casting by artificial neural networks approach
  70. Investigation of mechanical activation effect on high-volume natural pozzolanic cements
  71. In vitro: Anti-coccidia activity of Calotropis procera leaf extract on Eimeria papillata oocysts sporulation and sporozoite
  72. Determination of oil composition of cowpea (Vigna unguiculata L.) seeds under influence of organic fertilizer forms
  73. Activated partial thromboplastin time maybe associated with the prognosis of papillary thyroid carcinoma
  74. Treatment of rat brain ischemia model by NSCs-polymer scaffold transplantation
  75. Lead and cadmium removal with native yeast from coastal wetlands
  76. Characterization of electroless Ni-coated Fe–Co composite using powder metallurgy
  77. Ferrate synthesis using NaOCl and its application for dye removal
  78. Antioxidant, antidiabetic, and anticholinesterase potential of Chenopodium murale L. extracts using in vitro and in vivo approaches
  79. Study on essential oil, antioxidant activity, anti-human prostate cancer effects, and induction of apoptosis by Equisetum arvense
  80. Experimental study on turning machine with permanent magnetic cutting tool
  81. Numerical simulation and mathematical modeling of the casting process for pearlitic spheroidal graphite cast iron
  82. Design, synthesis, and cytotoxicity evaluation of novel thiophene, pyrimidine, pyridazine, and pyridine: Griseofulvin heterocyclic extension derivatives
  83. Isolation and identification of promising antibiotic-producing bacteria
  84. Ultrasonic-induced reversible blood–brain barrier opening: Safety evaluation into the cellular level
  85. Evaluation of phytochemical and antioxidant potential of various extracts from traditionally used medicinal plants of Pakistan
  86. Effect of calcium lactate in standard diet on selected markers of oxidative stress and inflammation in ovariectomized rats
  87. Identification of crucial salivary proteins/genes and pathways involved in pathogenesis of temporomandibular disorders
  88. Zirconium-modified attapulgite was used for removing of Cr(vi) in aqueous solution
  89. The stress distribution of different types of restorative materials in primary molar
  90. Reducing surface heat loss in steam boilers
  91. Deformation behavior and formability of friction stir processed DP600 steel
  92. Synthesis and characterization of bismuth oxide/commercial activated carbon composite for battery anode
  93. Phytochemical analysis of Ziziphus jujube leaf at different foliar ages based on widely targeted metabolomics
  94. Effects of in ovo injection of black cumin (Nigella sativa) extract on hatching performance of broiler eggs
  95. Separation and evaluation of potential antioxidant, analgesic, and anti-inflammatory activities of limonene-rich essential oils from Citrus sinensis (L.)
  96. Bioactivity of a polyhydroxy gorgostane steroid from Xenia umbellata
  97. BiCAM-based automated scoring system for digital logic circuit diagrams
  98. Analysis of standard systems with solar monitoring systems
  99. Structural and spectroscopic properties of voriconazole and fluconazole – Experimental and theoretical studies
  100. New plant resistance inducers based on polyamines
  101. Experimental investigation of single-lap bolted and bolted/bonded (hybrid) joints of polymeric plates
  102. Investigation of inlet air pressure and evaporative cooling of four different cogeneration cycles
  103. Review Articles
  104. Comprehensive review on synthesis, physicochemical properties, and application of activated carbon from the Arecaceae plants for enhanced wastewater treatment
  105. Research progress on speciation analysis of arsenic in traditional Chinese medicine
  106. Recent modified air-assisted liquid–liquid microextraction applications for medicines and organic compounds in various samples: A review
  107. An insight on Vietnamese bio-waste materials as activated carbon precursors for multiple applications in environmental protection
  108. Antimicrobial activities of the extracts and secondary metabolites from Clausena genus – A review
  109. Bioremediation of organic/heavy metal contaminants by mixed cultures of microorganisms: A review
  110. Sonodynamic therapy for breast cancer: A literature review
  111. Recent progress of amino acid transporters as a novel antitumor target
  112. Aconitum coreanum Rapaics: Botany, traditional uses, phytochemistry, pharmacology, and toxicology
  113. Corrigendum
  114. Corrigendum to “Petrology and geochemistry of multiphase post-granitic dikes: A case study from the Gabal Serbal area, Southwestern Sinai, Egypt”
  115. Corrigendum to “Design of a Robust sliding mode controller for bioreactor cultures in overflow metabolism via an interdisciplinary approach”
  116. Corrigendum to “Statistical analysis on the radiological assessment and geochemical studies of granite rocks in the north of Um Taghir area, Eastern Desert, Egypt”
  117. Corrigendum to “Aroma components of tobacco powder from different producing areas based on gas chromatography ion mobility spectrometry”
  118. Corrigendum to “Mechanical properties, elastic moduli, transmission factors, and gamma-ray-shielding performances of Bi2O3–P2O5–B2O3–V2O5 quaternary glass system”
  119. Erratum
  120. Erratum to “Copper(ii) complexes supported by modified azo-based ligands: Nucleic acid binding and molecular docking studies”
  121. Special Issue on Applied Biochemistry and Biotechnology (ABB 2021)
  122. Study of solidification and stabilization of heavy metals by passivators in heavy metal-contaminated soil
  123. Human health risk assessment and distribution of VOCs in a chemical site, Weinan, China
  124. Preparation and characterization of Sparassis latifolia β-glucan microcapsules
  125. Special Issue on the Conference of Energy, Fuels, Environment 2020
  126. Improving the thermal performance of existing buildings in light of the requirements of the EU directive 2010/31/EU in Poland
  127. Special Issue on Ethnobotanical, Phytochemical and Biological Investigation of Medicinal Plants
  128. Study of plant resources with ethnomedicinal relevance from district Bagh, Azad Jammu and Kashmir, Pakistan
  129. Studies on the chemical composition of plants used in traditional medicine in Congo
  130. Special Issue on Applied Chemistry in Agriculture and Food Science
  131. Strip spraying technology for precise herbicide application in carrot fields
  132. Special Issue on Pharmacology and Metabolomics of Ethnobotanical and Herbal Medicine
  133. Phytochemical profiling, antibacterial and antioxidant properties of Crocus sativus flower: A comparison between tepals and stigmas
  134. Antioxidant and antimicrobial properties of polyphenolics from Withania adpressa (Coss.) Batt. against selected drug-resistant bacterial strains
  135. Integrating network pharmacology and molecular docking to explore the potential mechanism of Xinguan No. 3 in the treatment of COVID-19
  136. Chemical composition and in vitro and in vivo biological assortment of fixed oil extracted from Ficus benghalensis L.
  137. A review of the pharmacological activities and protective effects of Inonotus obliquus triterpenoids in kidney diseases
  138. Ethnopharmacological study of medicinal plants in Kastamonu province (Türkiye)
  139. Protective effects of asperuloside against cyclophosphamide-induced urotoxicity and hematotoxicity in rats
  140. Special Issue on Essential Oil, Extraction, Phytochemistry, Advances, and Application
  141. Identification of volatile compounds and antioxidant, antibacterial, and antifungal properties against drug-resistant microbes of essential oils from the leaves of Mentha rotundifolia var. apodysa Briq. (Lamiaceae)
  142. Phenolic contents, anticancer, antioxidant, and antimicrobial capacities of MeOH extract from the aerial parts of Trema orientalis plant
  143. Chemical composition and antimicrobial activity of essential oils from Mentha pulegium and Rosmarinus officinalis against multidrug-resistant microbes and their acute toxicity study
  144. Special Issue on Marine Environmental Sciences and Significance of the Multidisciplinary Approaches
  145. An insightful overview of the distribution pattern of polycyclic aromatic hydrocarbon in the marine sediments of the Red Sea
  146. Antifungal–antiproliferative norcycloartane-type triterpenes from the Red Sea green alga Tydemania expeditionis
  147. Solvent effect, dipole moment, and DFT studies of multi donor–acceptor type pyridine derivative
  148. An extensive assessment on the distribution pattern of organic contaminants in the aerosols samples in the Middle East
  149. Special Issue on 4th IC3PE
  150. Energetics of carboxylic acid–pyridine heterosynthon revisited: A computational study of intermolecular hydrogen bond domination on phenylacetic acid–nicotinamide cocrystals
  151. A review: Silver–zinc oxide nanoparticles – organoclay-reinforced chitosan bionanocomposites for food packaging
  152. Green synthesis of magnetic activated carbon from peanut shells functionalized with TiO2 photocatalyst for Batik liquid waste treatment
  153. Coagulation activity of liquid extraction of Leucaena leucocephala and Sesbania grandiflora on the removal of turbidity
  154. Hydrocracking optimization of palm oil over NiMoO4/activated carbon catalyst to produce biogasoline and kerosine
  155. Special Issue on Pharmacology and metabolomics of ethnobotanical and herbal medicine
  156. Cynarin inhibits PDGF-BB-induced proliferation and activation in hepatic stellate cells through PPARγ
  157. Special Issue on The 1st Malaysia International Conference on Nanotechnology & Catalysis (MICNC2021)
  158. Surfactant evaluation for enhanced oil recovery: Phase behavior and interfacial tension
  159. Topical Issue on phytochemicals, biological and toxicological analysis of aromatic medicinal plants
  160. Phytochemical analysis of leaves and stems of Physalis alkekengi L. (Solanaceae)
  161. Phytochemical and pharmacological profiling of Trewia nudiflora Linn. leaf extract deciphers therapeutic potentials against thrombosis, arthritis, helminths, and insects
  162. Pergularia tomentosa coupled with selenium nanoparticles salvaged lead acetate-induced redox imbalance, inflammation, apoptosis, and disruption of neurotransmission in rats’ brain
  163. Protective effect of Allium atroviolaceum-synthesized SeNPs on aluminum-induced brain damage in mice
  164. Mechanism study of Cordyceps sinensis alleviates renal ischemia–reperfusion injury
  165. Plant-derived bisbenzylisoquinoline alkaloid tetrandrine prevents human podocyte injury by regulating the miR-150-5p/NPHS1 axis
  166. Network pharmacology combined with molecular docking to explore the anti-osteoporosis mechanisms of β-ecdysone derived from medicinal plants
  167. Chinese medicinal plant Polygonum cuspidatum ameliorates silicosis via suppressing the Wnt/β-catenin pathway
  168. Special Issue on Advanced Nanomaterials for Energy, Environmental and Biological Applications - Part I
  169. Investigation of improved optical and conductivity properties of poly(methyl methacrylate)–MXenes (PMMA–MXenes) nanocomposite thin films for optoelectronic applications
  170. Special Issue on Applied Biochemistry and Biotechnology (ABB 2022)
  171. Model predictive control for precision irrigation of a Quinoa crop
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