Neuroprotective Effects of Meloxicam and Selegiline in Scopolamine-Induced Cognitive Impairment and Oxidative Stress

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by a gradual decline in memory associated with shrinkage of brain tissue, with localized loss of neurons mainly in the hippocampus and basal forebrain, with diminished level of central cholinergic neurotransmitter-acetylcholine and also reported to be associated with accumulation of ubiquitinated proteins in neuronal inclusions and also with signs of inflammation. In these disorders, the abnormal protein aggregates may themselves trigger the expression of inflammatory mediators, such as cyclooxygenase 2 (COX-2). In the present study, the effects of Meloxicam, Selegiline, and coadministration of these drugs on scopolamine-induced learning and memory impairments in mice were investigated. Rectangular maze test, Morris water maze test, Locomotor activity, and Pole climbing test were conducted to evaluate the learning and memory parameters. Various biochemical parameters such as acetylcholinesterase(AChE), TBARS assay, catalase activity, and DPPH assay were also assessed. The present study demonstrates that Meloxicam, Selegiline, and co-administration of these test drugs had potential therapeutic effects on improving the antiamnesic activity in mice through inhibiting lipid peroxidation, augmenting endogenous antioxidant enzymes, and decreasing acetylcholinesterase activity in brain. The memory enhancing capacity of the drugs was very significant when compared to disease control (P < 0.001)

Optimizing the Treatment of Experimental Autoimmune Encephalomyelitis via Pulmonary Delivery of Soluble Antigen Arrays

Soluble antigen arrays (SAgAs) were used to treat experimental autoimmune encephalomyelitis (EAE), which is a mouse model for multiple sclerosis (MS). SAgAs offer a targeted therapy for MS, which is not present in current therapies. The different routes of administration, injection volume, dosing amount, and dosing schedule were explored to find that pulmonary instillation of SAgAs at 50 μL on a 200 nMol PLP basis on days 4, 7, and 10 of the study are most efficacious. With the next study, the different components of SAgAs (hyaluronic acid, PLP, LABL, and bifunctional peptide inhibitor) were also investigated via pulmonary delivery to find that PLP and SAgAs decreased the symptoms of EAE the most. PLP, HA-PLP, and SAgAs were further explored either via subcutaneous injection or pulmonary instillation to find that HA-PLP and SAgAs decreased disease progression in mice with EAE. Cytokine panels were also used to determine if tolerance was induced in these mice via the measurements of pro- and anti-inflammatory cytokines. Lastly, lung histology was explored to find signs of inflammation. Overall, SAgAs are found to be very efficacious in treating mice via PI and the presence of PLP makes a big difference. Other methods like the presence of regulatory T cells need to be used to find additional signs of tolerance induction in future studies

IDENTIFICATION OF NOVEL CYTOTOXIC PROSTAGLANDIN METABOLITES PRODUCED IN ARACHIDONOYL ETHANOLAMIDE-TREATED TUMOREGENIC KERATINOCYTES

Arachidonoyl ethanolamide (AEA) induces apoptosis in mouse tumorigenic keratinocytes (JWF-2 cells). Our previous data show that AEA is metabolized by COX-2 to pro-apoptotic J-series prostaglandins. COX-2 is an enzyme that is abundant in tumor cells but not in the normal epithelial cells surrounding the tumor. Thus, the pro-apoptotic J-series prostaglandins should be selectively synthesized in AEA-exposed tumor cells with elevated COX-2 expression. As such, AEA could be developed as a topical agent to treat non-melanoma skin cancer. The main goal of this project is to identify the specific J-series prostaglandins that are produced as a result of the metabolism of AEA by COX-2 using mass spectrometry. (ELISA analysis can detect J-series family prostaglandins but cannot distinguish between the individual J-series isoforms). Exogenous J-series prostaglandins were added to fresh cell culture medium, and the prostaglandins were extracted using solid phase extraction. Concentrated samples were then subjected to Liquid Chromatography/Electrospray Ionization /Mass Spectrometry (LC-ESI-MS) in negative mode for identification of J-series prostaglandin isoforms. Our data show good recovery of extracted species and acceptable resolution of these chemically similar standards. The AEA-treated cell culture medium and its control were extracted using the validated extraction protocol and analyzed using the method developed with LC-ESI-MS. The mass spectrum of AEA-treated, extracted and concentrated cell culture media clearly shows peaks at m/z ratio identified as the parent ion peak (M-H)� of ethanolamide conjugates of the J-series prostaglandins. The identification of the ethanolamide conjugates was confirmed by performing tandem mass spectrometry. It was also observed that with increased AEA concentration, the mass spectral intensity of the ethanolamide conjugates of J-series prostaglandins increased. The effect of COX-2 inhibitor and N-acetyl cysteine on the production of ethanolamide conjugates of J-series was also studied. The identification of the cytotoxic ethanolamide conjugates of J-series prostaglandins as the metabolites of AEA synthesized in tumor cells help us to determine the mechanism by which AEA induces apoptosis.M.S

Genetic divergence studies for yield and quality traits in high protein landraces of rice (Oryza sativa L.)

The present study was undertaken to study the extent of genetic diversity in high protein rice landraces with respect to yield, yield components and quality characters. In this direction, 30 high protein rice landraces, collected from different parts of country by ICAR-Indian Institute of Rice Research (ICAR-IIRR), Hyderabad along with the high protein check, CR DHAN 310 were evaluated during Kharif 2021 at ICAR-IIRR farm located at International Crops Research Institute of Semi Arid Tropics (ICRISAT), Hyderabad. The study examined the genetic divergence of high protein rice cultures for yield, quality and nutritional parameters. Multivariate analysis techniques of Mahalanobis D2 and Principal Component Analysis (PCA) were used to estimate the genetic diversity in the experimental material. In Mahalanobis D2, the 31 high protein rice cultures were divided into six clusters. Cluster I had highest number of rice cultures (19), followed by Cluster III and V with five, four cultures, respectively. The clusters, II, IV, VI were mono-genotypic. It was discovered that grouping of these cultures into several clusters was random and was not related to geographical diversity. Inter-cluster distances between clusters V and VI were maximum. Cluster V had also exhibited higher intra-cluster distance. Further, Cluster VI had showed maximum yield plant-1, grains per panicle-1, zinc content and test weight, while, Cluster V had recorded high protein content. The greatest contribution to genetic divergence was recorded by yield plant-1 (21.60%), followed by iron (10.54%) and zinc content (9.54%). In Principal Component Analysis, the first five Principal Components (PCs) with eigen values &gt;1 accounted for cumulative contribution of 67.69% to the total variability. The three traits, yield plant-1, iron content, and amylose content contributed the most to variability. The 2D scatter diagram exhibited 18 different clusters, out of which 11 clusters were mono-genotypic. Mahalanobis D2 Statistic and PCA concluded maximum genetic diversity between the landraces, JAK 248-3 and JAK 638 with JAK 611

Shift in Conformational Equilibrium Underlies the Oscillatory Phosphoryl Transfer Reaction in the Circadian Clock

Oscillatory phosphorylation/dephosphorylation can be commonly found in a biological system as a means of signal transduction though its pivotal presence in the workings of circadian clocks has drawn significant interest: for example in a significant portion of the physiology of Synechococcus elongatus PCC 7942. The biological oscillatory reaction in the cyanobacterial circadian clock can be visualized through its reconstitution in a test tube by mixing three proteins—KaiA, KaiB and KaiC—with adenosine triphosphate and magnesium ions. Surprisingly, the oscillatory phosphorylation/dephosphorylation of the hexameric KaiC takes place spontaneously and almost indefinitely in a test tube as long as ATP is present. This autonomous post-translational modification is tightly regulated by the conformational change of the C-terminal peptide of KaiC called the “A-loop” between the exposed and the buried states, a process induced by the time-course binding events of KaiA and KaiB to KaiC. There are three putative hydrogen-bond forming residues of the A-loop that are important for stabilizing its buried conformation. Substituting the residues with alanine enabled us to observe KaiB’s role in dephosphorylating hyperphosphorylated KaiC, independent of KaiA’s effect. We found a novel role of KaiB that its binding to KaiC induces the A-loop toward its buried conformation, which in turn activates the autodephosphorylation of KaiC. In addition to its traditional role of sequestering KaiA, KaiB’s binding contributes to the robustness of cyclic KaiC phosphorylation by inhibiting it during the dephosphorylation phase, effectively shifting the equilibrium toward the correct phase of the clock

The Differences of Funding in Education in the United States

Many individuals are disadvantaged due to the variation of funding in the United States. However, over time, schools have received a greater amount of funding to source primary through secondary schools in order to decrease the monetary gap between schools. In order to help those who do not have the same privileges, the government has increased funding, which has illustrated great strides in educational systems. Students are now able to receive the materials and supplies necessary for an effective learning environment. For example, students now have experienced teachers, as well as textbooks to allow them to learn curriculum. As the funding for education has increased in schools over time, an increase in test scores display the help that funding has provided to less fortunate districts. There have been trends over time describing places that have lesser funding throughout schools in their district, such as South Chicago neighborhoods, not allowing students to work up from these neighborhoods. To provide these students with opportunity, lesser funding over the years have been mapped out and traced to specific school districts. With this funding, students are able to excel and reach for opportunities they may not have been able to with previous resources. Throughout the states, there has also been evidence that funding in education from local governments have varied based on representation. In order for students to obtain the necessary materials for an effective education, tracing the allowance for funds in each state and changes over time in these states to permit students to reach higher success is beneficial. Therefore, through the understanding of trends for funding in states over time, governments and funds can be targeted towards these less fortunate educational systems to provide these students with opportunities for success, as well

Synthesis, characterization and antimicrobial activity of a series of substituted coumarin-3-carboxylatosilver(1) complexes

A series of new coumarin-derived carboxylate ligands and their silver(I) complexes have been synthesized, characterized and screened for their in vitro antibacterial activity against a range of Gram-positive and Gram-negative bacteria as well as for their antifungal activity against a clinical isolate of Candida albicans. The ligands were synthesised by either acid or base hydrolysis of their corresponding esters, which in turn were synthesised via the Knoevenegal reaction. The reaction of silver(I) nitrate with the coumarin carboxylate ligands in either aqueous or aqueous/ethanol solutions allowed the isolation of a series of novel Ag(I) carboxylate complexes. Whilst none of the ligands showed any antimicrobial activity, a number of the Ag(I) complexes exhibited potent activity. In particular, Ag(I) complexes of hydroxy-substituted coumarin carboxylates demonstrated potent activity against the clinically important methicillin-resistant Staphylococcus aureus (MRSA) bacterium (MIC80 = 0.63 μM)

Mechanism of action of coumarin and silver(I)- coumarin complexes against the pathogenic yeast Candida albicans

The anti-fungal activity and mode of action of a range of silver(I)- coumarin complexes was examined. The most potent silver(I)- coumarin complexes, namely 7-hydroxycoumarin-3-carboxylatosilver(I), 6-hydroxycoumarin-3-carboxylatosilver(I) and 4-oxy-3-nitrocoumarinbis(1,10-phenanthroline)silver(I), had MIC80 values of between 69.1 and 4.6 M against the pathogenic yeast Candida albicans. These compounds also reduced respiration, lowered the ergosterol content of cells and increased the trans-membrane leakage of amino acids. A number of the complexes disrupted cytochrome synthesis in the cell and induced the appearance of morphological features consistent with cell death by apoptosis. These compounds appear to act by disrupting the synthesis of cytochromes which directly aVects the cell's ability to respire. A reduction in respiration leads to a depletion in ergosterol biosynthesis and a consequent disruption of the integrity of the cell membrane. Disruption of cytochrome biosynthesis may induce the onset of apoptosis which has been shown previously to be triggered by alteration in the location of cytochrome c. Silver(I)- coumarin complexes demonstrate good anti-fungal activity and manifest a mode of action distinct to that of the conventional azole and polyene drugs thus raising the possibility of their use when resistance to conventional drug has emerged or in combination with such drugs

Impact of Molecular Weight on Lymphatic Drainage of a Biopolymer-Based Imaging Agent

New lymphatic imaging technologies are needed to better assess immune function and cancer progression and treatment. Lymphatic uptake depends mainly on particle size (10–100 nm) and charge. The size of carriers for imaging and drug delivery can be optimized to maximize lymphatic uptake, localize chemotherapy to lymphatic metastases, and enable visualization of treatment deposition. Toward this end, female BALB/c mice were injected subcutaneously in the hind footpad or forearm with a series of six different molecular weight hyaluronan (HA) near-infrared dye (HA-IR820) conjugates (ca. 5–200 nm). Mice were imaged using whole body fluorescent imaging over two weeks. HA-IR820 fluorescence was clearly visualized in the draining lymphatic capillaries, and in the popliteal and iliac or axillary lymph nodes. The 74-kDa HA-IR820 had the largest lymph node area-under-the-curve. In contrast to prior reports, mice bearing limb tumors exhibited three-fold longer retention of 74-kDa HA-IR820 in the popliteal node compared to mice without tumors. HA conjugate kinetics and disposition can be specifically tailored by altering their molecular weight. The specific lymphatic uptake and increased nodal retention of HA conjugates indicate significant potential for development as a natural biopolymer for intralymphatic drug delivery and imaging

Role of cell cycle events and apoptosis in mediating the anti-cancer activity of a silver(I) complex of 4-hydroxy-3-nitro-coumarin-bis(phenanthroline) in human malignant cancer cells.

The central objective of the current study was to investigate the potential in vitro anti-proliferative effect of 4-hydroxy-3-nitro-coumarin (hncH), and the mixed-ligand silver (I) complex of 4-oxy-3-nitro-coumarin-bis (phenanthroline), [Ag(hnc)(phen)2] using four human-derived model cell lines. In addition, selected mechanistic studies were carried out using the most sensitive of the four cell lines. Results obtained show that the complex could decrease the proliferation of all four cell lines including neoplastic renal and hepatic, namely A-498 and HepG2 cells, respectively, along with two non-neoplastic renal and hepatic cell lines, HK-2 and Chang, respectively. Furthermore, non-neoplastic hepatic cells (Chang) appeared to be less sensitive to the effect of the complex, but this effect was not replicated in the non-neoplastic renal (HK-2) cells. Based on IC50 values [Ag(hnc)(phen)2] was shown to be almost four times more potent than cisplatin, using HepG2 cells. In addition, the observed anti-proliferative effect was shown to be both dose- and time-dependent. Furthermore, the complex was shown to decrease DNA synthesis, but did not intercalate with it. Moreover, there was no evidence that P-glycoprotein-mediated multi-drug resistance was likely to decrease antiproliferative activity. Cytological stains, analysis of genomic DNA, and biochemical assays [caspase-3 and -9 and cleaved poly(ADP-ribose)-polymerase protein] showed that cell death appeared to result from apoptosis, with the possibility of secondary necrosis. Additionally, flow cytometric analysis showed that the complex functioned through an alteration in cell cycle progression. Taken together, [Ag(hnc)(phen)2] has been shown to be a more potent anti-proliferative agent than cisplatin, capable of altering key biochemical events leading to cell death. Additional mechanistic studies are underway to probe more fully its mechanism of action