Peptide Carriers to Improve Uptake and Functionality and to Cross BBB to Arrest Secondary Injury Post TBI

Cell-penetrating peptides (CPP) possess the ability to transport different cargos efficiently across the cell membrane. In this study, the ability of a CPP to cross the blood-brain barrier (BBB) and stably carry therapeutic components across the plasma membrane was assessed. The therapeutic efficacies were determined by the ability of the KAFAK conjugated anti-inflammatory peptide drug to inhibit the progression of secondary neuronal damage in a traumatic brain injury (TBI) model at 7-day post-injury (DPI) by inhibiting the production of inflammatory cytokines. A related study with minocycline and PgP-rolipram was conducted to improve the outcome in TBI models at 14 DPI. Similarly, the ability of halloysite nanotubules (HNTs) to cross the BBB was investigated. Improving the BBB-crossing carrier library is the primary objective of this study, which seeks to identify reliable carrier molecules that can transport therapeutic compounds to the brain to treat brain injury and disorders. Initially, two peptides were synthesized: (1) an anti-inflammatory peptide (AIP-1) that specifically target the MK2 pathway (mitogen-activated protein kinase) to regulate the inflammatory response (control), and (2) KAFAK, a CPP conjugated to AIP-1. The intracellular delivery, therapeutic efficacy, and cytotoxicity of these peptides were compared in rodent primary BBB cells (primary brain microvascular endothelial cells (BMVEC\u27s)) and four cell lines (macrophages (RAW, J774), neuronal cells (SHEP-1), and hepatocytes (HEP G2)). KAFAK did not induce cell toxicity at concentrations below 250 µM in primary BMVEC cells and below 1000 µM in the cell lines. Subsequently, the CPP conjugate was validated in vivo for its uptake and localization in the brain and its therapeutic efficacy in a TBI mouse model following non-invasive intranasal administration for six days. The results showed that KAFAK was primarily localized in the olfactory bulbs. Some diffused throughout the cortex, and it reduced cytokine (IL-1β, IL-6, and TNF-α) production in TBI mice as compared to vehicle-treated mice. Halloysite nanotubules (HNTs), naturally available nanoparticles, were also explored for their ability to penetrate the BBB. HNTs were loaded with rhodamine isothiocyanate (RITC) to determine their uptake and localization in the brain at 4, 24, and 48 hours after a single noninvasive intranasal administration. Another group of mice received HNTs loaded with diazepam intranasally for six days and was evaluated for behavioral changes versus mice that received HNTs alone. Fluorescence from RITC was observed in the brain tissue of mice treated intranasally with HNTs loaded with RITC but not in mice treated with RITC alone. The intensity of fluorescence decreased over time, and no HNTs-associated behavioral changes were observed. Mice treated with HNT-diazepam exhibited diazepam-associated behavioral changes, indicating that HNTs could penetrate the BBB and release the drug into the brain. In summary, HNTs and CPPs have demonstrated that they have the potential to transport drugs across the BBB

Molecular cloning and characterization of regulatory enzymes in threonine biosynthetic pathway from soybean

The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file.Title from title screen of research.pdf file (viewed on May 12, 2009)Includes bibliographical references.Thesis (M.S.) University of Missouri-Columbia 2007.Dissertations, Academic -- University of Missouri--Columbia -- Agronomy.Soybeans (Glycine max [L.] Merr.) are a good source of protein and oil. Despite being a very good source of protein, soybeans nutritional value is limited by low proportion of sulfur amino acids methionine and cysteine demanding supplementation of soy-based animal feed with synthetic amino acids. The cost involved aside, other essential amino acids, called second-tier amino acids, are also becoming limiting in animal feed due to this practice. Threonine is one of those second-tier amino acids and improving its proportion in soybean seeds can improve the nutritional value of soy-based animal feed. The manipulation of regulatory enzymes in the biosynthesis pathway of this amino acid is one method used to increase the proportion of free threonine in soybean seeds. One prerequisite for such an approach is the isolation and characterization of the genes encoding the key enzymes. In this study, the genes coding both homoserine dehydrogenase (HSDH) and threonine synthase (TS), two regulatory enzymes in threonine biosynthesis pathway, have been isolated and named gmhsdh and gmts, respectively. The gmhsdh seems to code a mono-functional, cytosolic HSDH and is interrupted by 11 introns and hence, the cDNA encoding HSDH was also isolated. The gmts encodes a chloroplast localized TS and this gene has no introns. Multiple sequence analysis of the HSDH amino acid sequence leads to a proposition that this protein is feedback insensitive to threonine. Verification of this fact could make this a very good candidate for achieving a favorable proportion of threonine in the free amino acid pools of soybean seeds

Evaluation of Anti-Ulcer and Anti-Inflammatory Activity of Ethanolic Extract of Solanum Pubescens Willd Leaves on Experimental Animals.

Herbs are staging a comeback and herbal ‘renaissance’ is happening all over the globe. The herbal products today symbolise safety in contrast to the synthetics that are regarded as unsafe to human and environment. Although herbs had been priced for their medicinal, flavoring and aromatic qualities for centuries, the synthetic products of the modern age surpassed their importance, for a while. However, the blind dependence on synthetics is over and people are returning to the naturals with hope of safety and security. Over three-quarters of the world population relies mainly on plants and plant extract for health care. More than 30% of the entire plant species, at one time or other were used for medicinal purposes the study of the ethonolic extract of Solanum pubescens willd leaves possess Anti-ulcer and Anti-inflammatory activity in animal models. Solanum pubescens willd showed a significant decrease in the ulcer development in both the animal models ( pylorus ligated model and Aspirin-induced ulcer model) used in the study

From Inaccuracy to Insight: Identifying Medication Discrepancies through Observational Reconciliation at a Tertiary Care Hospital, Bhimavaram of Andhra Pradesh

Background: Medication reconciliation is the process of examining the patient's entire medication regimen at the time of admission, transfer, and discharge and comparing it with the regimen being considered for the new setting of care. This helps to prevent unintentional inconsistencies across transitions in medical care. Medication reconciliation protects patients from medication side effects while ensuring that they receive standard care. It serves as the baseline from which therapeutic interventions are developed, drug treatment is continued upon admission, and self-care is continued upon release. Objectives: Determining the frequency and kinds of discrepancies discovered during medication reconciliation was the main goal of this study. Determining the effect of medication reconciliation to assess the possible seriousness of medication inconsistencies and ascertain the drug's role in medication errors was the secondary goal. Methodology: In the inpatient units of a tertiary care hospital in the West Godavari District, a prospective, observational study on medication reconciliation was conducted for six months. Results: Of the 385 patients that made up this study, 224 (58.18%) were males and 161 (41.8%) were females. In 169 (43.89%) of the patients, medication discrepancies were detected. There were inconsistencies discovered at several transition points: 50 disparities were detected at admission, 50 during the transfer phase, and 17 on the discharge. Conclusion: A multi-centric assessment including parameters like the percentage of inpatients encountering at least one major medication error would be intriguing. This may support the idea that drug reconciliation is crucial for patient safety

Understanding The Role Of Health Literacy In Self-Medication: Findings From A Cross-Sectional Study In West Godavari District Of Andhra Pradesh

Background: Self-medication is the practice of treating any ailment or symptom that a person diagnoses for themselves without first visiting a physician. Different communities display different behaviours; hence the purpose of this study is to statistically investigate the patterns and prevalence of self-medication usage. Although health literacy practices have been increasingly recommended in public health literature, there is a lack of studies that examine the relationships between health literacy and self-medication. Methodology: A quantitative, descriptive, cross-sectional, community-based research approach was used in a sample of 316 participants. Health literacy was measured by Single Item Literacy Screener. Data was analysed using SPSS 29.0 version. Results:  A total of 316 participants agreed to participate (63.9% were females). The results showed that more than half, 53.4% had adequate health literacy. The prevalence of self-medication was 74%, in these 52% had used medicines by previous prescription and 8% used alternative medicine. There was a significant relationship between the overall health literacy level and practice of self-medication. Conclusion: Improving the health literacy level of the public can reduce inappropriate self-medication Therefore, the design and implementation of training programs are necessary to increase the perception on the risk of self-medication. Appropriate reading skills are important for accessing health information, using health care services, and achieving desirable health outcomes

Prion Disease and the Innate Immune System

Prion diseases or transmissible spongiform encephalopathies are a unique category of infectious protein-misfolding neurodegenerative disorders. Hypothesized to be caused by misfolding of the cellular prion protein these disorders possess an infectious quality that thrives in immune-competent hosts. While much has been discovered about the routing and critical components involved in the peripheral pathogenesis of these agents there are still many aspects to be discovered. Research into this area has been extensive as it represents a major target for therapeutic intervention within this group of diseases. The main focus of pathological damage in these diseases occurs within the central nervous system. Cells of the innate immune system have been proven to be critical players in the initial pathogenesis of prion disease, and may have a role in the pathological progression of disease. Understanding how prions interact with the host innate immune system may provide us with natural pathways and mechanisms to combat these diseases prior to their neuroinvasive stage. We present here a review of the current knowledge regarding the role of the innate immune system in prion pathogenesis

MLH1 mediates PARP-dependent cell death in response to the methylating agent N-methyl-N-nitrosourea

Background:Methylating agents such as N-methyl-N-nitrosourea (MNU) can cause cell cycle arrest and death either via caspase-dependent apoptosis or via a poly(ADP-ribose) polymerase (PARP)-dependent form of apoptosis. We wished to investigate the possible role of MLH1 in signalling cell death through PARP.Methods:Fibroblasts are particularly dependent on a PARP-mediated cell death response to methylating agents. We used hTERT-immortalised normal human fibroblasts (WT) to generate isogenic MLH1-depleted cells, confirmed by quantitative PCR and western blotting. Drug resistance was measured by clonogenic and cell viability assays and effects on the cell cycle by cell sorting. Damage signalling was additionally investigated using immunostaining.Results:MLH1-depleted cells were more resistant to MNU, as expected. Despite having an intact G2/M checkpoint, the WT cells did not initially undergo cell cycle arrest but instead triggered cell death directly by PARP overactivation and nuclear translocation of apoptosis-inducing factor (AIF). The MLH1-depleted cells showed defects in this pathway, with decreased staining for phosphorylated H2AX, altered PARP activity and reduced AIF translocation. Inhibitors of PARP, but not of caspases, blocked AIF translocation and greatly decreased short-term cell death in both WT and MLH1-depleted cells. This MLH1-dependent response to MNU was not blocked by inhibitors of ATM/ATR or p53.Conclusion:These novel data indicate an important role for MLH1 in signalling PARP-dependent cell death in response to the methylating agent MNU

Gαq-containing G proteins regulate B cell selection and survival and are required to prevent B cell–dependent autoimmunity

Survival of mature B cells is regulated by B cell receptor and BAFFR-dependent signals. We show that B cells from mice lacking the Gαq subunit of trimeric G proteins (Gnaq−/− mice) have an intrinsic survival advantage over normal B cells, even in the absence of BAFF. Gnaq−/− B cells develop normally in the bone marrow but inappropriately survive peripheral tolerance checkpoints, leading to the accumulation of transitional, marginal zone, and follicular B cells, many of which are autoreactive. Gnaq−/− chimeric mice rapidly develop arthritis as well as other manifestations of systemic autoimmune disease. Importantly, we demonstrate that the development of the autoreactive B cell compartment is the result of an intrinsic defect in Gnaq−/− B cells, resulting in the aberrant activation of the prosurvival factor Akt. Together, these data show for the first time that signaling through trimeric G proteins is critically important for maintaining control of peripheral B cell tolerance induction and repressing autoimmunity