Assessment of Phytochemical Analysis,Nutritional Composition and Antimicrobial Activity of<i> Moringa oleifera</i>

Moringa oleifera is a miracle plant rich in nutrients, antioxidants, and antibiotic properties. Present study was designed to evaluate various biochemical attributes of leaves and flowers of M. oleifera. Plant parts (leaves, flowers) of M. oleifera, collected from different roadsides of Multan district, Punjab, Pakistan, were used as experimental material. Result indicates that alkaloids, saponin, carbohydrates, fats, and protein had a high value in the aqueous extract of both leaves and flowers of M. oleifera. Whereas phenol content was high in methanolic leaves extract and the phenol contents were high in aqueous extract of flowers. The extract yield of M. oleifera leaves and flowers both showed a higher percentage in aqueous extract (57.5%), followed by methanol extract and lowest in ethyl acetate extract. Flavonoids contents were higher in ethyl acetate extract of leaves (33.67%) and aqueous extract of flowers (53.71%). While crude fiber was high in methanolic extract of leaves (12.40%) and in flowers crude fiber was high in ethyl acetate extract (15.86%). The moisture contents were higher in leaves (8.87%) than flowers (7.3%) and similarly, ash percentage in flowers (52.60%) than leaves (41.84%). Ethyl acetate extracts of M. oleifera leaves show antibacterial activity against Pseudomonas aeruginosa while methanolic extract of M. oleifera flowers shows antibacterial activity against Xanthomonas sp. Maximum growth inhibits show in all extracts of leaves against Aspergillus flavus, F. oxysporum, and P. glabrum except for the concentrated aqueous extract of leaves. While in flowers maximum growth inhibits all extracts against P. glabrum, A. niger, and A. flavus except the diluted ethyl acetate extract. Phytochemicals present in different parts of moringa have significant edible and commercial potential. Moringa extracts exhibited significant antimicrobial activity, therefore have applications in pharmaceuticals.     

Kidney Dysfunction in Adult Offspring Exposed In Utero to Type 1 Diabetes Is Associated with Alterations in Genome-Wide DNA Methylation

Background

Fetal exposure to hyperglycemia impacts negatively kidney development and function.

Objective

Our objective was to determine whether fetal exposure to moderate hyperglycemia is associated with epigenetic alterations in DNA methylation in peripheral blood cells and whether those alterations are related to impaired kidney function in adult offspring.

Design

Twenty nine adult, non-diabetic offspring of mothers with type 1 diabetes (T1D) (case group) were matched with 28 offspring of T1D fathers (control group) for the study of their leukocyte genome-wide DNA methylation profile (27,578 CpG sites, Human Methylation 27 BeadChip, Illumina Infinium). In a subset of 19 cases and 18 controls, we assessed renal vascular development by measuring Glomerular Filtration Rate (GFR) and Effective Renal Plasma Flow (ERPF) at baseline and during vasodilatation produced by amino acid infusion.

Results

Globally, DNA was under-methylated in cases vs. controls. Among the 87 CpG sites differently methylated, 74 sites were less methylated and 13 sites more methylated in cases vs. controls. None of these CpG sites were located on a gene known to be directly involved in kidney development and/or function. However, the gene encoding DNA methyltransferase 1 (DNMT1)—a key enzyme involved in gene expression during early development–was under-methylated in cases. The average methylation of the 74 under-methylated sites differently correlated with GFR in cases and controls.

Conclusion

Alterations in methylation profile imprinted by the hyperglycemic milieu of T1D mothers during fetal development may impact kidney function in adult offspring. The involved pathways seem to be a nonspecific imprinting process rather than specific to kidney development or function.     

A critical balance between Cyclin B synthesis and Myt1 activity controls meiosis entry in Xenopus oocytes

In fully grown oocytes, meiosis is arrested at first prophase until species-specific initiation signals trigger maturation. Meiotic resumption universally involves early activation of M phase-promoting factor (Cdc2 kinase-Cyclin B complex, MPF) by dephosphorylation of the inhibitory Thr14/Tyr15 sites of Cdc2. However, underlying mechanisms vary. In Xenopus oocytes, deciphering the intervening chain of events has been hampered by a sensitive amplification loop involving Cdc2-Cyclin B, the inhibitory kinase Myt1 and the activating phosphatase Cdc25. In this study we provide evidence that the critical event in meiotic resumption is a change in the balance between inhibitory Myt1 activity and Cyclin B neosynthesis. First, we show that in fully grown oocytes Myt1 is essential for maintaining prophase I arrest. Second, we demonstrate that, upon upregulation of Cyclin B synthesis in response to progesterone, rapid inactivating phosphorylation of Myt1 occurs, mediated by Cdc2 and without any significant contribution of Mos/MAPK or Plx1. We propose a model in which the appearance of active MPF complexes following increased Cyclin B synthesis causes Myt1 inhibition, upstream of the MPF/Cdc25 amplification loop.     

Energy‐Restricted High‐Fat Diets Only Partially Improve Markers of Systemic and Adipose Tissue Inflammation

This study aimed at investigating whether the weight loss due to energy‐restricted high‐fat diets is accompanied with parallel improvements in metabolic markers and adipose tissue inflammation. Eight‐week‐old C57BL/6J mice were given free access to a low‐fat (LF) or a high‐fat (45% of energy from fat—HF) diet for 6 months. Restricting intake of the HF diet by 30% (HFR) during the last 2 months of the HF feeding trial decreased fasting plasma insulin, homeostasis model assessment of insulin resistance (HOMA_IR), and plasma triglyceride levels and improved hepatic steatosis compared to ad libitum HF feeding, indicating an improved metabolic profile. Further, analysis of gonadal white adipose tissue (GWAT) gene expression by microarray and quantitative PCR analyses demonstrated that HFR downregulated expression of genes linked to cell and focal adhesion, cytokine‐cytokine receptor interaction, and endoplasmic reticulum (ER)–associated degradation pathway. However, HFR had no effect on circulating plasminogen activator inhibitor‐1 (PAI‐1) and nonesterified fatty acid levels, which were persistently higher in both HF and HFR groups compared to the LF group. Furthermore, HFR had a negative effect on plasma total adiponectin level. Finally, while HFR decreased GWAT monocyte chemotactic protein‐1 (MCP‐1), interleukin‐2 (IL‐2), and PAI‐1 levels, it did not affect several other cytokines including granulocyte‐macrophage colony‐stimulating factor, interferon‐γ, IL‐1β, IL‐6, and IL‐10. In summary, energy‐restricted high‐fat diets improve insulin sensitivity, while only partially improving markers of systemic and adipose tissue inflammation. In conclusion, our study supports the recommended low‐fat intake for overall cardiovascular health.     

Characterization of lipoteichoic acid structures from three probiotic Bacillus strains: involvement of d-alanine in their biological activity

Probiotics represent a potential strategy to influence the host’s immune system thereby modulating immune response. Lipoteichoic Acid (LTA) is a major immune-stimulating component of Gram-positive cell envelopes. This amphiphilic polymer, anchored in the cytoplasmic membrane by means of its glycolipid component, typically consists of a poly (glycerol-phosphate) chain with d-alanine and/or glycosyl substitutions. LTA is known to stimulate macrophages in vitro, leading to secretion of inflammatory mediators such as Nitric Oxide (NO). This study investigates the structure–activity relationship of purified LTA from three probiotic Bacillus strains (Bacillus cereus CH, Bacillus subtilis CU1 and Bacillus clausii O/C). LTAs were extracted from bacterial cultures and purified. Chemical modification by means of hydrolysis at pH 8.5 was performed to remove d-alanine. The molecular structure of native and modified LTAs was determined by ¹H NMR and GC–MS, and their inflammatory potential investigated by measuring NO production by RAW 264.7 macrophages. Structural analysis revealed several differences between the newly characterized LTAs, mainly relating to their d-alanylation rates and poly (glycerol-phosphate) chain length. We observed induction of NO production by LTAs from B. subtilis and B. clausii, whereas weaker NO production was observed with B. cereus. LTA dealanylation abrogated NO production independently of the glycolipid component, suggesting that immunomodulatory potential depends on d-alanine substitutions. d-alanine may control the spatial configuration of LTAs and their recognition by cell receptors. Knowledge of molecular mechanisms behind the immunomodulatory abilities of probiotics is essential to optimize their use.