Vitamin D-binding protein deficiency: an underrecognized Mendelian disorder of vitamin D metabolism.

Vitamin D-binding protein (VDBP) deficiency is a recently discovered apparently benign biochemical disorder that can masquerade as treatment-resistant vitamin D deficiency and is likely underrecognized. We present the case of a child with persistently low 25OH vitamin D levels despite replacement therapy. Exome sequencing revealed a novel homozygous nonsense variant in the GC gene, leading to undetectable levels of VDBP. Interestingly, exome sequencing also revealed a homozygous loss-of-function variant in ZNF142, which likely explains the additional clinical features of recurrent febrile convulsions and global developmental delay. Our findings corroborate the two previously reported patients with autosomal recessive VDBP deficiency caused by biallelic GC variants and emphasize the importance of measuring VDBP levels in cases of apparent vitamin D deficiency that is treatment-resistant. We also urge caution in concluding "atypical" presentations without careful investigation of a potential dual molecular diagnosis.

Cloning of CAT genes in Satsuma mandarin and their expression characteristics in response to environmental stress and arbuscular mycorrhizal fungi.

KEY MESSAGE: CitCAT1 and CitCAT2 were cloned and highly expressed in mature leaves. High temperatures up-regulated CitCAT1 expression, while low temperatures and Diversispora versiformis up-regulated CitCAT2 expression, maintaining a low oxidative damage. Catalase (CAT), a tetrameric heme-containing enzyme, removes hydrogen peroxide (H2O2) to maintain low oxidative damage in plants exposed to environmental stress. This study aimed to clone CAT genes from Citrus sinensis cv. "Oita 4" and analyze their expression patterns in response to environmental stress, exogenous abscisic acid (ABA), and arbuscular mycorrhizal fungal inoculation. Two CAT genes, CitCAT1 (NCBI accession: PP067858) and CitCAT2 (NCBI accession: PP061394) were cloned, and the open reading frames of their proteins were 1479 bp and 1539 bp, respectively, each encoding 492 and 512 amino acids predicted to be localized in the peroxisome, with CitCAT1 being a stable hydrophilic protein and CitCAT2 being an unstable hydrophilic protein. The similarity of their amino acid sequences reached 83.24%, and the two genes were distantly related. Both genes were expressed in stems, leaves, flowers, and fruits, accompanied by the highest expression in mature leaves. In addition, CitCAT1 expression was mainly up-regulated by high temperatures (37 °C), exogenous ABA, and PEG stress within a short period of time, whereas CitCAT2 expression was up-regulated by exogenous ABA and low-temperature (4 °C) stress. Low temperatures (0 °C) for 12 h just up-regulated CitCAT2 expression in Diversispora versiformis-inoculated plants, and D. versiformis inoculation up-regulated CitCAT2 expression, along with lower hydrogen peroxide and malondialdehyde levels in mycorrhizal plants at low temperatures. It is concluded that CitCAT2 has an important role in resistance to low temperatures as well as mycorrhizal enhancement of host resistance to low temperatures.

Association of Polymorphisms in PD-1 and LAG-3 Genes with Acute Myeloid Leukemia.

Background and objectives: Acute myeloid leukemia (AML) is a hematological malignancy characterized by uncontrolled proliferation of immature myeloid cells. Immune checkpoint molecules such as programmed cell death protein 1 (PD-1) and lymphocyte activation gene-3 (LAG-3) are essential for controlling anti-tumor immune responses. This study aims to explore the correlation between specific genetic variations (SNPs) in the PDCD1 (rs2227981) and LAG3 (rs12313899) genes and the likelihood of developing AML in the Saudi population. Material and methods: total of 98 Saudi AML patients and 131 healthy controls were genotyped for the PDCD1 rs2227981 and LAG3 rs12313899 polymorphisms using TaqMan genotyping assays. A logistic regression analysis was conducted to evaluate the relationship between the SNPs and AML risk using several genetic models. Results: The results revealed a significant association between the PDCD1 rs2227981 polymorphism and increased AML risk. In AML patients, the frequency of the G allele was considerably greater than in healthy controls (OR = 1.93, 95% CI: 1.31-2.81, p = 0.00080). The GG and AG genotypes were associated with a very high risk of developing AML (p < 0.0001). In contrast, no significant association was observed between the LAG3 rs12313899 polymorphism and AML risk in the studied population. In silico analysis of gene expression profiles from public databases suggested the potential impact of PDCD1 expression levels on the overall survival of AML patients. Conclusions: This study provides evidence for the association of the PDCD1 rs2227981 polymorphism with an increased risk for AML in the Saudi population.

Insights into the Effect of Chitosan and ß-Cyclodextrin Hybridization of Zeolite-A on Its Physicochemical and Cytotoxic Properties as a Bio-Carrier for 5-Fluorouracil: Equilibrium and Release Kinetics Studies.

Synthetic zeolite-A (ZA) was hybridized with two different biopolymers (chitosan and ß-cyclodextrin) producing biocompatible chitosan/zeolite-A (CS/ZA) and ß-cyclodextrin/zeolite-A (CD/ZA) biocomposites. The synthetic composites were assessed as bio-carriers of the 5-fluorouracil drug (5-Fu) with enhanced properties, highlighting the impact of the polymer type. The hybridization by the two biopolymers resulted in notable increases in the 5-Fu loading capacities, to 218.2 mg/g (CS/ZA) and 291.3 mg/g (CD/ZA), as compared to ZA (134.2 mg/g). The loading behaviors using ZA as well as CS/ZA and CD/ZA were illustrated based on the classic kinetics properties of pseudo-first-order kinetics (R2 > 0.95) and the traditional Langmuir isotherm (R2 = 0.99). CD/ZA shows a significantly higher active site density (102.7 mg/g) in comparison to CS/ZA (64 mg/g) and ZA (35.8 mg/g). The number of loaded 5-Fu per site of ZA, CS/ZA, and CD/ZA (>1) validates the vertical ordering of the loaded drug ions by multi-molecular processes. These processes are mainly physical mechanisms based on the determined Gaussian energy (<8 kJ/mol) and loading energy (<40 kJ/mol). Both the CS/ZA and CD/ZA 5-Fu release activities display continuous and controlled profiles up to 80 h, with CD/ZA exhibiting much faster release. According to the release kinetics studies, the release processes contain non-Fickian transport release properties, suggesting cooperative diffusion and erosion release mechanisms. The cytotoxicity of 5-Fu is also significantly enhanced by these carriers: 5-Fu/ZA (11.72% cell viability), 5-Fu/CS/ZA (5.43% cell viability), and 5-Fu/CD/ZA (1.83% cell viability).

Synthesis and Characterization of Methoxy-Exfoliated Montmorillonite Nanosheets as Potential Carriers of 5-Fluorouracil Drug with Enhanced Loading, Release, and Cytotoxicity Properties.

Natural bentonite clay (BE) underwent modification steps that involved the exfoliation of its layers into separated nanosheets (EXBE) and further functionalization of these sheets with methanol, forming methoxy-exfoliated bentonite (Mth/EXBE). The synthetically modified products were investigated as enhanced carriers of 5-fluorouracil as compared to raw bentonite. The modification process strongly induced loading properties that increased to 214.4 mg/g (EXBE) and 282.6 mg/g (Mth/EXBE) instead of 124.9 mg/g for bentonite. The loading behaviors were illustrated based on the kinetic (pseudo-first-order model), classic isotherm (Langmuir model), and advanced isotherm modeling (monolayer model of one energy). The Mth/EBE carrier displays significantly higher loading site density (95.9 mg/g) as compared to EXBE (66.2 mg/g) and BE (44.9 mg/g). The loading numbers of 5-Fu in each site of BE, EXBE, and Mth/EXBE (>1) reflect the vertical orientation of these loaded ions involving multi-molecular processes. The loading processes that occurred appeared to be controlled by complex physical and weak chemical mechanisms, considering both Gaussian energy (<8 KJ/mol) as well as loading energy (<40 KJ/mol). The releasing patterns of EXBE and Mth/EXBE exhibit prolonged and continuous properties up to 100 h, with Mth/EXBE displaying much faster behaviors. Based on the release kinetic modeling, the release reactions exhibit non-Fickian transport release properties, validating cooperative diffusion and erosion release mechanisms. The cytotoxicity of 5-Fu is also significantly enhanced by these carriers: 5-Fu/BE (8.6% cell viability), 5-Fu/EXBE (2.21% cell viability), and 5-Fu/Mth/EXBE (0.73% cell viability).

Insight into the Morphological Properties of Nano-Kaolinite (Nanoscrolls and Nanosheets) on Its Qualification as Delivery Structure of Oxaliplatin: Loading, Release, and Kinetic Studies.

Natural kaolinite underwent advanced morphological-modification processes that involved exfoliation of its layers into separated single nanosheets (KNs) and scrolled nanoparticles as nanotubes (KNTs). Synthetic nanostructures have been characterized as advanced and effective oxaliplatin-medication (OXAP) delivery systems. The morphological-transformation processes resulted in a remarkable enhancement in the loading capacity to 304.9 mg/g (KNs) and 473 mg/g (KNTs) instead of 29.6 mg/g for raw kaolinite. The loading reactions that occurred by KNs and KNTs displayed classic pseudo-first-order kinetics (R2 > 0.90) and conventional Langmuir isotherms (R2 = 0.99). KNTs exhibit a higher active site density (80.8 mg/g) in comparison to KNs (66.3 mg/g) and raw kaolinite (6.5 mg/g). Furthermore, compared to KNs and raw kaolinite, each site on the surface of KNTs may hold up to six molecules of OXAP (n = 5.8), in comparison with five molecules for KNs. This was accomplished by multi-molecular processes, including physical mechanisms considering both the Gaussian energy (<8 KJ/mol) and the loading energy (<40 KJ/mol). The release activity of OXAP from KNs and KNTs exhibits continuous and regulated profiles up to 100 h, either by KNs or KNTs, with substantially faster characteristics for KNTs. Based on the release kinetic investigations, the release processes have non-Fickian transport-release features, indicating cooperative-diffusion and erosion-release mechanisms. The synthesized structures have a significant cytotoxicity impact on HCT-116 cancer cell lines (KNs (71.4% cell viability and 143.6 g/mL IC-50); KNTs (11.3% cell viability and 114.3 g/mL IC-50). Additionally, these carriers dramatically increase OXAP's cytotoxicity (2.04% cell viability, 15.4 g/mL IC-50 (OXAP/KNs); 0.6% cell viability, 4.5 g/mL IC-50 (OXAP/KNTs)).

Polyphenols-Rich Extract of Calotropis procera Alone and in Combination with Trichoderma Culture Filtrate for Biocontrol of Cantaloupe Wilt and Root Rot Fungi.

Fungal diseases have always been a major problem for cantaloupe crops; however, synthetic fungicides are hazardous to humans and the environment. Consequently, a feasible alternative to fungicides without side effects could be by using bio agents and naturally occurring plants with antibacterial potential. This study has achieved a novel procedure for managing wilt and root rot diseases by potentially using Trichoderma sp. culture filtrates in consortium with plant extract of Calotropis procera, Rhizoctonia solani, Fusarium oxysporum, and Pythium ultimum, which were isolated from infected cantaloupe roots with identified root rot symptoms. The antagonistic activity of four Trichoderma isolates and analysis of antibiotics and filtrate enzymes of the most active Trichoderma isolate were determined as well as phytochemical analysis of C. procera plant extract using HPLC-UV. The obtained results showed that all Trichoderma isolates considerably lowered the radial growth of P. ultimum, R. solani, and F. oxysporum in varying degrees. The scanning electron micrographs illustrate the mycoparasitic nature of Trichoderma sp. on F. oxysporum. The phytochemical analysis of C. procera indicated that phenolic contents were the major compounds found in extracts, such as vanillin (46.79%), chlorogenic acid (30.24%), gallic acid (8.06%), and daidzein (3.45%) but including only a low amount of the flavonoid compounds rutin, naringenin, and hesperetin. The Pot experiment's findings showed that cantaloupe was best protected against wilting and root rot diseases when it was treated with both Trichoderma sp. culture filtrates (10%) and C. procera extract of (15 mg/mL), both alone and in combination. This study demonstrates that the application of bio agent Trichoderma spp. filtrate with C. procera phenol extract appears useful for controlling wilting and root rot disease in cantaloupe. This innovative approach could be used as an alternative to chemical fungicide for the control of wilting and rot root diseases.

Medication use and medical history of 155 patients with oral lichenoid lesions: a retrospective study.

Several medications have been reported as possible etiologic factors for oral lichen planus (OLP) and oral lichenoid lesions (OLLs). This study investigated the medication profile and medical history of patients with biopsy-proven OLP or OLLs, also classified by the clinically nonspecific term oral lichenoid mucositis (OLM), in a busy oral medicine clinic. The University of Florida College of Dentistry records from 2009 to 2014 were searched retrospectively for all patients with a biopsy-proven diagnosis of OLP, OLLs, or OLM. Patients were excluded if dysplasia or carcinoma was diagnosed concurrently at the same biopsy site. The demographics, clinical parameters, systemic diseases, histologic diagnosis, and direct immunofluorescence testing results were recorded. Medication category use was recorded based on both commonly used medications and those that have been reportedly linked to lichenoid disease in the literature. A total of 155 patients with an average age of 63.6 years were included. The majority of patients were women (76.8%) and Caucasian (91.8%). Most of the lesions were multifocal and mixed (white-red) in appearance. The most common systemic conditions were hypertension (n = 80; 51.6%) followed by thyroid disease (n = 52; 33.5%) and diabetes (n = 26; 16.8%). Antihypertensives were the most common medication category followed by, in descending order, nonsteroidal anti-inflammatory drugs, cholesterol-lowering medications, psychiatric medications, and thyroid replacement drugs. The records revealed that 87.7% of the patients took at least 1 medication from 1 of the categories studied. Medication use is common in patients with biopsy-proven OLP or OLLs. Although causation cannot be assessed from the results of this study, the clinician should consider the possibility of medication as a complicating factor in patients with OLP or OLLs.

TIM3 and CTLA4 immune checkpoint polymorphisms are associated with acute myeloid leukemia in Saudi Arabia.

BACKGROUND: Immune checkpoints are receptors on the surface of T cells that function crucially in suppressing the immune response, and they are implicated in autoimmunity and cancer diseases. AIM: The present study aimed to investigate the relationship between functional single nucleotide polymorphisms (SNPs) of two immune checkpoint molecules, CTLA-4 and TIM-3, and acute myeloid leukemia (AML) in a Saudi population. METHODS: Two SNPs in CTLA-4 (rs231775, A > G) and TIM-3 (rs10515746, A > C) were genotyped in 229 subjects, including 98 patients and 131 healthy controls, from the Saudi population using TaqMan assay methods. Differential expression of these two genes was performed using in silico analysis. RESULTS: An association was found between polymorphisms in TIM-3 (OR: 6.01; 95% CI: 3.99-9.05, P < 0.0001) and the risk of AML. Inversely, the rs231775 SNP in the CTLA-4 gene was found to protect against AML in allelic, dominant, and additive models (P < 0.05). A significantly higher expression of TIM-3 in the blood of individuals with AML was observed. CONCLUSION: This is the first study focusing on single nucleotide polymorphisms (SNPs) for CTLA-4 and TIM-3 in acute myeloid leukemia patients in a Saudi community and could be a potential new prognostic factor for this disease.

Endophytic Bacillus vallismortis and Bacillus tequilensis bacteria isolated from medicinal plants enhance phosphorus acquisition and fortify Brassica napus L. vegetative growth and metabolic content.

Phosphorus fertilization imposes critical limitations on crop productivity and soil health. The aim of the present work is to explore the potential of two phosphate solubilizing bacteria (PSB) species in phosphorus supplementation of canola (Brassica napus L.). Out of 38 bacterial isolates obtained from nine medicinal plants, two bacterial strains (20P and 28P) were proved as the most potent for the in-vitro tricalcium phosphate solubilization test. These isolates verified their activity toward different enzymes as nitrogenase and alkaline phosphatase. Also, 20P and 28P gave a high amount of indole-3-acetic acid, 34.16 µg/ml and 35.20 µg/ml, respectively, and were positive for siderophores production as they detected moderate affinity for iron chelation. Molecular identification confirmed that strain 20P was Bacillus vallismortis and strain 28P was Bacillus tequilensis. A pot experiment was conducted to study the effect of four different phosphorus concentrations (0%, 50%, 75%, and 100% P) each alone and/or in combination with B. vallismortis, B. tequilensis, or both bacterial isolates on the vegetative growth and some physiological parameters of canola. The combined treatment of 50% phosphorus + (B. vallismortis + B. tequilensis) was generally the most effective with respect to shoot height, shoot dry mass, leaf area, photosynthetic pigment fractions, total sugar content, and accumulated NPK content. In contrast, the rhizosphere pH reached the minimum value under the same treatment. These findings highlighted the potential use of PSB (B. vallismortis and B. tequilensis) along with phosphorus fertilization as a safe sustainable tactic.

Hypomorphic variants of SEL1L-HRD1 ER-associated degradation are associated with neurodevelopmental disorders.

Recent studies using cell type-specific knockout mouse models have improved our understanding of the pathophysiological relevance of suppressor of lin-12-like-HMG-CoA reductase degradation 1 (SEL1L-HRD1) endoplasmic reticulum-associated (ER-associated) degradation (ERAD); however, its importance in humans remains unclear, as no disease variant has been identified. Here, we report the identification of 3 biallelic missense variants of SEL1L and HRD1 (or SYVN1) in 6 children from 3 independent families presenting with developmental delay, intellectual disability, microcephaly, facial dysmorphisms, hypotonia, and/or ataxia. These SEL1L (p.Gly585Asp, p.Met528Arg) and HRD1 (p.Pro398Leu) variants were hypomorphic and impaired ERAD function at distinct steps of ERAD, including substrate recruitment (SEL1L p.Gly585Asp), SEL1L-HRD1 complex formation (SEL1L p.Met528Arg), and HRD1 activity (HRD1 p.Pro398Leu). Our study not only provides insights into the structure-function relationship of SEL1L-HRD1 ERAD, but also establishes the importance of SEL1L-HRD1 ERAD in humans.

The immunopathological landscape of human pre-TCRα deficiency: From rare to common variants.

We describe humans with rare biallelic loss-of-function PTCRA variants impairing pre-α T cell receptor (pre-TCRα) expression. Low circulating naive αß T cell counts at birth persisted over time, with normal memory αß and high γδ T cell counts. Their TCRα repertoire was biased, which suggests that noncanonical thymic differentiation pathways can rescue αß T cell development. Only a minority of these individuals were sick, with infection, lymphoproliferation, and/or autoimmunity. We also report that 1 in 4000 individuals from the Middle East and South Asia are homozygous for a common hypomorphic PTCRA variant. They had normal circulating naive αß T cell counts but high γδ T cell counts. Although residual pre-TCRα expression drove the differentiation of more αß T cells, autoimmune conditions were more frequent in these patients compared with the general population.

Immunohistochemical Evaluation of the Pathological Effects of Diabetes Mellitus on the Major Salivary Glands of Albino Rats.

OBJECTIVES: Diabetes mellitus is a notorious chronic disease characterized by hyperglycemia. Our study aimed to determine the expression of cytokeratin 17 (CK17) in all major salivary glands of diabetic albino rats to provide more information about the pathological effects of DM on the intracellular structures of the gland parenchyma. MATERIALS AND METHODS: Twenty male adult albino rats were utilized in the experiment and divided into two equal groups, group 1 (control rats) and group 2 (diabetic rats). The animals were sacrificed 45 days after diabetes induction. The major salivary gland complex of all groups was dissected and prepared for evaluation by histological and immunohistochemical expression of CK17. RESULTS: Histological results prove that the salivary gland parenchyma of diabetic group undergo gland atrophy characterized with the presence of degenerated acini, dilated duct system, and presence of duct-like structure with predominance of fibrous tissue compartment and discrete fat cells. Immunohistochemical expression of CK17 of major salivary gland of control group revealed negative to diffuse mild expression in all duct cells and some serous acinar cells, whereas mucous acini were negatively stained. On the other hand, major salivary gland parenchyma of diabetic group demonstrated mild to strong expression of duct cells more concentrated at their apical part with moderate to strong expression of some serous acini of diffuse type, whereas mucous acini of both submandibular gland and sublingual gland were negatively stained. CONCLUSION: The severity and prevalence of CK 17 in our results are predictive of the pathological influence of the DM that interferes with saliva production and/or secretion leading to dry mouth. The results also showed clear changes in the cytokeratin expression of diabetic sublingual salivary gland, although it had little effect in the routine histological study with hematoxylin and eosin, confirming that routine studies are not sufficient to form a definitive opinion.

Comparative Study of Cytokeratin Immunostaining of Parotid Gland Parenchyma in Normal, Diabetic, and Excretory Duct Ligation of Mongrel Dogs.

OBJECTIVES: The present study aimed to give a glimpse of the normal distribution of intermediate filaments within the parotid gland parenchyma of mongrel dogs and to reveal the pathological changes that may occur as a result of the effects of diabetes mellitus or atrophy of the gland caused by the ligation of the excretory duct to discover whether there is a similarity in these pathological behaviors. MATERIALS AND METHODS: Twelve healthy mongrel dogs were used in the experiment and were divided into three groups: group I (the control group), group II (dogs with alloxan-induced diabetes), and group III (dogs with the right-side duct-ligated parotid gland). The dogs were sacrificed 45 days after the parotid excretory duct were tied. The right parotid gland of all groups was dissected and prepared for histological and immunohistochemical expression of cytokeratin 17 assay. RESULTS: Histological findings confirmed that the parotid gland parenchyma of the diabetic group had glandular atrophy characterized by the loss of gland structure, degenerated acini, and dilatation of the duct system. Moreover, there is a predominance of the fibrous component with the presence of fat cells within the gland compartments. On the contrary, the excretory duct-ligated group undergoes severe glandular atrophy of the previous character with the presence of duct-like structure as well as extravasation and vasodilatation. Immunohistochemical expression of cytokeratin 17 in control parotid using an immunoperoxidase technique showed that cytokeratin expression varies from negative to mild in all ducts and some serous acinar cells. The gland parenchyma of the diabetic group showed mild to strong cytokeratin expression of duct cells more concentrated in the apical part with moderate to strong expression of diffuse type in some serous acini. The intensity of cytokeratin 17 in gland compartments of the excretory duct-ligated group revealed a variation in expression that ranged from negative to strong diffuse staining throughout the gland. CONCLUSION: The severity and prevalence of cytokeratin 17 in our results are predictive of the pathological influence of both diabetes mellitus and duct ligation on the cytokeratin intracellular filaments of the salivary gland parenchyma in a different way that interferes with saliva production and/or secretion leading to xerostomia.

Evaluation of the level of integration of the dental curriculum in Umm Al-Qura University.

Objectives: To identify the models and levels of integration of the undergraduate dental curriculum in Umm Al-Qura University in KSA. This comprehensive evaluation and analysis of the current dental curriculum will facilitate better planning for curriculum reform, thus improving the quality of dental education. Methods: All courses were evaluated by three reviewers who independently checked the most recent course specifications forms (2021) to extract data relating to course descriptions and contents. A model of integration was identified for each course (using a modified Harden's integration ladder). Courses and their relative weighting (by credit hours) were mapped to the level of integration by years, departments, course classification, and educational methods. The overall pattern of curriculum integration was then determined. Results: All courses exhibited some level of integration to varying degrees throughout years and across departments. The most frequently used model is the nested model of integration. The overall pattern of curriculum integration is low to moderate. Highly integrated courses are only taught during the second and final years and are managed by the Departments of Basic Oral Sciences and Restorative Dentistry. Clinical courses represent 44.3% of the curriculum although only 26.6% of clinical courses have a high level of integration. Problem-based learning/case-based learning (PBL/CBL) and clinical training strategies are mostly applied in moderately to highly integrated courses, although PBL/CBL is the least used educational method throughout the curriculum. Conclusion: All courses exhibited some level of integration with an overall low to moderate pattern. More collaborative planning and working between departments are recommended to increase the level of integration of courses throughout different academic years. In addition, modern educational strategies such as PBL/CBL and blended learning should be implemented more in our dental curriculum.

Effect of Biotic Elicitors on the Growth, Antioxidant Activity and Metabolites Accumulation in In Vitro Propagated Shoots of Pueraria tuberosa.

Pueraria tuberosa contains a wide range of bioactive compounds, including polyphenols, alkaloids, and phytosterols, which make it valuable to the pharmaceutical and food industries. Elicitor compounds trigger the defense mechanisms in plants and are widely used to increase the yield of bioactive molecules in in vitro cultures. The present study was conducted to evaluate the effects of different concentrations of biotic elicitors such as yeast extract (YE), pectin (PEC), and alginate (ALG) on growth, antioxidant activity, and metabolite accumulation in in vitro propagated shoots of P. tuberosa. The elicitors applied to shoot cultures of P. tuberosa significantly increased biomass (shoot number, fresh weight, and dry weight), and metabolites such as protein, carbohydrates, chlorophyll, total phenol (TP), and total flavonoid (TF) contents, as well as antioxidant activity compared to untreated control. Biomass, TP, and TF contents, as well as antioxidant activity, were most significant in cultures treated with 100 mg/L PEC. In contrast, chlorophyll, protein, and carbohydrate increased most in cultures treated with 200 mg/L ALG. Application of 100 mg/L of PEC led to the accumulation of high amounts of isoflavonoids including puerarin (220.69 µg/g), daidzin (2935.55 µg/g), genistin (5612 µg/g), daidzein (479.81 µg/g), and biochanin-A (111.511 µg/g) as analyzed by high-performance liquid chromatography (HPLC). Total isoflavonoids content of 100 mg/L PEC treated shoots was obtained as 9359.56 µg/g, 1.68-fold higher than in vitro propagated shoots without elicitors (5573.13 µg/g) and 2.77-fold higher than shoots of the mother plant (3380.17 µg/g). The elicitor concentrations were optimized as 200 mg/L YE, 100 mg/L PEC, and 200 mg/L ALG. Overall, this study showed that the application of different biotic elicitors resulted in better growth, antioxidant activity, and accumulation of metabolites in P. tuberosa, which could lead to obtaining phytopharmaceutical advantages in the future.

Synthesis and Characterization of ß-Cyclodextrin-Hybridized Exfoliated Kaolinite Single Nanosheets as Potential Carriers of Oxaliplatin with Enhanced Loading, Release, and Cytotoxic Properties.

Natural kaolinite was subjected to a successful exfoliation process into separated kaolinite nanosheets (KNs), followed by hybridization with ß-cyclodextrin biopolymer (ß-CD), forming an advanced bio-nanocomposite (ß-CD/KNs). The synthetic products were evaluated as enhanced delivery structures for oxaliplatin chemotherapy (OXAPN). The hybridization of KNs with ß-CD polymer notably enhanced the loading capacity to 355.3 mg/g (ß-CD/KNs) as compared to 304.9 mg/g for KNs. The loading of OXAPN into both KNs and ß-CD/KNs displayed traditional pseudo-first-order kinetics (R2 > 0.85) and a conventional Langmuir isotherm (R2 = 0.99). The synthetic ß-CD/KNs validates a greater occupied effective site density (98.7 mg/g) than KNs (66.3 mg/g). Furthermore, the values of the n steric parameter (4.7 (KNs) and 3.6 (ß-CD/KNs)) reveal the vertical orientation of the loaded molecules and the loading of them by multi-molecular mechanisms. These mechanisms are mainly physical processes based on the obtained Gaussian energy (<8 KJ/mol) and loading energy (<40 KJ/mol). The release profiles of both KNs and ß-CD/KNs extend for about 120 h, with remarkably faster rates for ß-CD/KNs. According to the release kinetic findings, the release of OXAPN displays non-Fickian transport behavior involving the cooperation of diffusion and erosion mechanisms. The KNs and ß-CD/KNs as free particles showed considerable cytotoxicity and anticancer properties against HCT-116 cancer cell lines (71.4% cell viability (KNs) and 58.83% cell viability (ß-CD/KNs)). Additionally, both KNs and ß-CD/KNs significantly enhanced the OXAPN's cytotoxicity (2.04% cell viability (OXAPN/KNs) and 0.86% cell viability (OXAPN/ß-CD/KNs).

A Comparative Evaluation of Antibacterial Efficacy of Moringa oleifera Leaf Extract, Octenidine Dihydrochloride, and Sodium Hypochlorite as Intracanal Irrigants against Enterococcus faecalis: An In Vitro Study.

Objectives: The aim of this study is to investigate and compare the microbial efficacy of Moringa oleifera leaf extract, octenidine dihydrochloride (OCT), NaOCl, and their combinations as intracanal irrigants against Enterococcus faecalis. Materials and Methods: Sixty single-rooted mandibular premolars were decoronated followed by root canal preparation. Each root specimen was autoclaved, inoculated with E. faecalis, and incubated at 37°C for 48 hr. Then, the specimens were divided into six groups based on the irrigation solution used: 2.5% NaOCl (Group 1), 0.1% OCT (Group 2), M. oleifera leaves extract (Group 3), a combination of M. oleifera extract and 1.25% NaOCl (Group 4), a combination of M. oleifera extract and OCT (Group 5) and normal saline (Group 6). Microbial samples were taken from each root canal before (S1) and after (S2) irrigation and the bacterial viability was assessed using colony-forming units (CFU) on bile esculin agar plates. Results: Comparing the number of CFU/ml before and after irrigation showed a significant reduction (P < 0.001) in all studied groups. Comparison between the CFU/ml after irrigation by NaOCl and each of the combination groups showed a significant difference. Conclusion: M. oleifera leaves extract and 0.1% OCT solutions have antibacterial effect against E. faecalis comparable to 2.5% NaOCl and might be used as root canal irrigants. The combination groups showed better antimicrobial activities than individual irrigants. However, further studies are required to investigate the biocompatibility and possible toxic effects of the tested irrigants.

Wheat Grains as a Sustainable Source of Protein for Health.

Protein deficiency is recognized among the major global health issues with an underestimation of its importance. Genetic biofortification is a cost-effective and sustainable strategy to overcome global protein malnutrition. This study was designed to focus on protein-dense grains of wheat (Triticum aestivum L.) and identify the genes governing grain protein content (GPC) that improve end-use quality and in turn human health. Genome-wide association was applied using the 90k iSELECT Infinium and 35k Affymetrix arrays with GPC quantified by using a proteomic-based technique in 369 wheat genotypes over three field-year trials. The results showed significant natural variation among bread wheat genotypes that led to detecting 54 significant quantitative trait nucleotides (QTNs) surpassing the false discovery rate (FDR) threshold. These QTNs showed contrasting effects on GPC ranging from -0.50 to +0.54% that can be used for protein content improvement. Further bioinformatics analyses reported that these QTNs are genomically linked with 35 candidate genes showing high expression during grain development. The putative candidate genes have functions in the binding, remobilization, or transport of protein. For instance, the promising QTN AX-94727470 on chromosome 6B increases GPC by +0.47% and is physically located inside the gene TraesCS6B02G384500 annotated as Trehalose 6-phosphate phosphatase (T6P), which can be employed to improve grain protein quality. Our findings are valuable for the enhancement of protein content and end-use quality in one of the major daily food resources that ultimately improve human nutrition.