Fe65-engineered neuronal exosomes encapsulating corynoxine-B ameliorate cognition and pathology of Alzheimer's disease.

Alzheimer's disease (AD) is a neurodegenerative disorder characterized by the predominant impairment of neurons in the hippocampus and the formation of amyloid plaques, hyperphosphorylated tau protein, and neurofibrillary tangles in the brain. The overexpression of amyloid-ß precursor protein (APP) in an AD brain results in the binding of APP intracellular domain (AICD) to Fe65 protein via the C-terminal Fe65-PTB2 interaction, which then triggers the secretion of amyloid-ß and the consequent pathogenesis of AD. Apparently, targeting the interaction between APP and Fe65 can offer a promising therapeutic approach for AD. Recently, exosome, a type of extracellular vesicle with diameter around 30-200 nm, has gained much attention as a potential delivery tool for brain diseases, including AD, due to their ability to cross the blood-brain barrier, their efficient uptake by autologous cells, and their ability to be surface-modified with target-specific receptor ligands. Here, the engineering of hippocampus neuron cell-derived exosomes to overexpress Fe65, enabled the development of a novel exosome-based targeted drug delivery system, which carried Corynoxine-B (Cory-B, an autophagy inducer) to the APP overexpressed-neuron cells in the brain of AD mice. The Fe65-engineered HT22 hippocampus neuron cell-derived exosomes (Fe65-EXO) loaded with Cory-B (Fe65-EXO-Cory-B) hijacked the signaling and blocked the natural interaction between Fe65 and APP, enabling APP-targeted delivery of Cory-B. Notably, Fe65-EXO-Cory-B induced autophagy in APP-expressing neuronal cells, leading to amelioration of the cognitive decline and pathogenesis in AD mice, demonstrating the potential of Fe65-EXO-Cory-B as an effective therapeutic intervention for AD.

PPARɑ Ligand Caudatin Improves Cognitive Functions and Mitigates Alzheimer's Disease Defects By Inducing Autophagy in Mice Models.

The autophagy-lysosomal pathway (ALP) is a major cellular machinery involved in the clearance of aggregated proteins in Alzheimer disease (AD). However, ALP is dramatically impaired during AD pathogenesis via accumulation of toxic amyloid beta (Aß) and phosphorylated-Tau (phospho-Tau) proteins in the brain. Therefore, activation of ALP may prevent the increased production of Aß and phospho-Tau in AD. Peroxisome proliferator-activated receptor alpha (PPARα), a transcription factor that can activate autophagy, and transcriptionally regulate transcription factor EB (TFEB) which is a key regulator of ALP. This suggests that targeting PPARα, to reduce ALP impairment, could be a viable strategy for AD therapy. In this study, we investigated the anti-AD activity of Caudatin, an active constituent of Cynanchum otophyllum (a traditional Chinese medicinal herb, Qing Yang Shen; QYS). We found that Caudatin can bind to PPARα as a ligand and augment the expression of ALP in microglial cells and in the brain of 3XTg-AD mice model. Moreover, Caudatin could activate PPARα and transcriptionally regulates TFEB-augmented lysosomal degradation of Aß and phosphor-Tau aggregates in AD cell models. Oral administration of Caudatin decreased AD pathogenesis and ameliorated the cognitive dysfunction in 3XTg-AD mouse model. Conclusively, Caudatin can be a potential AD therapeutic agent via activation of PPARα-dependent ALP.

Studying the use of a proposed three-level grossing protocol in the histopathological diagnosis of COVID-19-associated rhinomaxillary mucormycosis.

Introduction: Mucormycosis is an acute and rapidly progressing opportunistic fungal infection. COVID-19-associated mucormycosis (CAM) had re-emerged as a complication of COVID-19 infection during the second wave of the pandemic in 2021. The rhinomaxillary form is a variant of the rhino-cerebral mucormycosis that presents a diagnostic challenge to the dentist and the oral and maxillofacial pathologist. Gross examination of pathological specimens is the most undermined step even though it plays a vital role in the final diagnosis. No studies have described this post-clinical step for the maxillofacial soft and hard tissue submitted for examination. Material and Methods: A prospective comparative study was carried out on 52 COVID-19-associated rhinomaxillary mucormycosis (CARM) cases to achieve complete, representative, and informative sampling of the submitted tissue and establish a three-level gross macroscopic examination protocol. Complete clinical and radiological histories were recorded after informed, written consent from every patient was received. Details of the number and type of samples received were recorded, grossing procedure was done as per the proposed three-level grossing protocol and were then compared to the presence of fungal hyphae in the soft tissue or decalcified hard tissue. Result: All 100% of the samples consisted of soft tissue (maxillary sinus lining), while 90.4% of the samples contained different hard tissue specimens. Seventy percent of the grossing workload was carried out by first-year oral pathology residents. Sixty-seven point three percent of the total soft tissue samples submitted showed no presence of fungal hyphae, while 69.2% of total decalcified sections of hard tissue were positive for fungal hyphae with a positive correlation. Out of the 29 cases grossed via the three-level grossing protocol, 89.6% of the cases were histopathologically positive for fungal hyphae. Thus a positive association (P < 0.05) between histopathological diagnosis and the proposed three-level grossing protocol was found. Conclusion: It is imperative to recognise that no mucormycosis report is to be signed out without multi-site (three-level grossed) bone decalcified reports. There is an immediate need to realise how vital documentation, correct laboratory practices, and grossing are for accurate histopathological diagnosis.

Delivery of Apoplastic Extracellular Vesicles Encapsulating Green-Synthesized Silver Nanoparticles to Treat Citrus Canker.

The citrus canker pathogen Xanthomonas axonopodis has caused severe damage to citrus crops worldwide, resulting in significant economic losses for the citrus industry. To address this, a green synthesis method was used to develop silver nanoparticles with the leaf extract of Phyllanthus niruri (GS-AgNP-LEPN). This method replaces the need for toxic reagents, as the LEPN acts as a reducing and capping agent. To further enhance their effectiveness, the GS-AgNP-LEPN were encapsulated in extracellular vesicles (EVs), nanovesicles with a diameter of approximately 30-1000 nm naturally released from different sources, including plant and mammalian cells, and found in the apoplastic fluid (APF) of leaves. When compared to a regular antibiotic (ampicillin), the delivery of APF-EV-GS-AgNP-LEPN and GS-AgNP-LEPN to X. axonopodis pv. was shown to have more significant antimicrobial activity. Our analysis showed the presence of phyllanthin and nirurinetin in the LEPN and found evidence that both could be responsible for antimicrobial activity against X. axonopodis pv. Ferredoxin-NADP+ reductase (FAD-FNR) and the effector protein XopAI play a crucial role in the survival and virulence of X. axonopodis pv. Our molecular docking studies showed that nirurinetin could bind to FAD-FNR and XopAI with high binding energies (-10.32 kcal/mol and -6.13 kcal/mol, respectively) as compared to phyllanthin (-6.42 kcal/mol and -2.93 kcal/mol, respectively), which was also supported by the western blot experiment. We conclude that (a) the hybrid of APF-EV and GS-NP could be an effective treatment for citrus canker, and (b) it works via the nirurinetin-dependent inhibition of FAD-FNR and XopAI in X. axonopodis pv.

Co-Occurrence of Taurodontism in Nonsyndromic Cleft Lip and Palate Patients in Subset of Indian Population: A Case-Control Study Using CBCT.

The present study aimed to assess the co-occurrence of taurodontism in nonsyndromic cleft lip and palate (NSCLP) patients in a subset of Indian population using cone beam computed tomography (CBCT).The study was a retrospective case-control kinds assessing 1500 CBCT scans over a period of 2 years; 67 scans out of 1500 showed cleft lip and palate (CLP). After fulfilling the inclusion and exclusion criteria, 38 subjects out of 67 comprised the case group; 80 randomly selected subjects constituted the control group. The first and second permanent molars in both the arches (8 teeth) of each subject were assessed for the presence and severity of taurodontism using the objective criteria. Statistical analysis was done using the χ2 test. Inter and intraobserver agreement was evaluated by Kappa statistics.71.05% subjects showed taurodontism in case group, while 45% subjects showed its presence in control group; results being significant for both the groups. However, the comparison of number of teeth with taurodontism in the 2 groups came out to be statistically insignificant. Also no significant association of taurodontism was seen with gender or the different types of cleft (P = .437). Hypotaurodontism was the most prevalent type.There was a higher prevalence of taurodontism in both groups which had been underestimated in the previous studies. This emphasizes the significance of the use of CBCT in detection of dental anomalies which would otherwise go undetected.

Peptide-Conjugated Nano Delivery Systems for Therapy and Diagnosis of Cancer.

Peptides are strings of approximately 2-50 amino acids, which have gained huge attention for theranostic applications in cancer research due to their various advantages including better biosafety, customizability, convenient process of synthesis, targeting ability via recognizing biological receptors on cancer cells, and better ability to penetrate cell membranes. The conjugation of peptides to the various nano delivery systems (NDS) has been found to provide an added benefit toward targeted delivery for cancer therapy. Moreover, the simultaneous delivery of peptide-conjugated NDS and nano probes has shown potential for the diagnosis of the malignant progression of cancer. In this review, various barriers hindering the targeting capacity of NDS are addressed, and various approaches for conjugating peptides and NDS have been discussed. Moreover, major peptide-based functionalized NDS targeting cancer-specific receptors have been considered, including the conjugation of peptides with extracellular vesicles, which are biological nanovesicles with promising ability for therapy and the diagnosis of cancer.

Factors Affecting Extracellular Vesicles Based Drug Delivery Systems.

Extracellular vesicles (EVs) play major roles in intracellular communication and participate in several biological functions in both normal and pathological conditions. Surface modification of EVs via various ligands, such as proteins, peptides, or aptamers, offers great potential as a means to achieve targeted delivery of therapeutic cargo, i.e., in drug delivery systems (DDS). This review summarizes recent studies pertaining to the development of EV-based DDS and its advantages compared to conventional nano drug delivery systems (NDDS). First, we compare liposomes and exosomes in terms of their distinct benefits in DDS. Second, we analyze what to consider for achieving better isolation, yield, and characterization of EVs for DDS. Third, we summarize different methods for the modification of surface of EVs, followed by discussion about different origins of EVs and their role in developing DDS. Next, several major methods for encapsulating therapeutic cargos in EVs have been summarized. Finally, we discuss key challenges and pose important open questions which warrant further investigation to develop more effective EV-based DDS.

In-silico Prediction of the Beta-carboline Alkaloids Harmine and Harmaline as Potent Drug Candidates for the Treatment of Parkinson's disease.

BACKGROUND: Parkinson's disease (PD) is a progressive neurodegenerative disease manifested by core symptoms of loss of motor control and postural instability. Loss of dopaminergic neurons is the cause of PD, thus enhancing dopamine level by pharmacological treatment is one of the key treatment strategies for PD. However, the limitations of current treatment strategies open the possibility of novel drug candidates for the treatment of PD. OBJECTIVE: To investigate the anti-PD potential of Harmine and Harmaline. We aim to evaluate the therapeutic potential of Harmine and Harmaline by in-silico approaches; molecular docking, pharmacokinetic and Prediction of Activity Spectra for Substances (PASS) analysis were used for evaluating the therapeutic potential of Harmine and Harmaline and standard drug levodopa (L-DOPA). METHODS: Auto dock vina was used for molecular docking of all three compounds against D2- and D3- dopamine receptors. The pharmacokinetics (PKs) and toxicity profile were predicted by pkCSM, and the pharmacological activity was predicted by PASS analysis. RESULTS: Molecular docking showed a higher binding affinity of Harmine and Harmaline as compared to L-DOPA, and these results were supported by in-silico pharmacokinetic and toxicity profiling. Moreover, PASS analysis showed anti-PD activity of Harmine and Harmaline. CONCLUSION: Harmine and Harmaline exhibit higher binding affinity towards D2- and D3- dopamine receptors compared to L-DOPA, and PKs and toxicity profile support their potential as drug candidates for PD therapy.

Synthesis, In-Vitro and In-Silico Evaluation of Silver Nanoparticles with Root Extract of Withania somnifera for Antibacterial Activity via Binding of Penicillin-Binding Protein-4.

BACKGROUND: Metal Nanoparticles (NPs) have been widely used for various applications in biomedical sciences, including in drug delivery, and as therapeutic agents, but limited owing to their toxicity towards the healthy tissue. This warrants an alternative method, which can achieve the desired activity with much reduced or no toxicity. Being a biological product, Withania somnifera (W. somnifera) is environment friendly, besides being less toxic as compared to metal-based NPs. However, the exact mechanism of action of W. somnifera for its antibacterial activities has not been studied so far. OBJECTIVE: To develop "silver nanoparticles with root extract of W. somnifera (AgNPs-REWS)" for antimicrobial and anticancer activities. Furthermore, the analysis of their mechanism of action will be studied. METHODS: Using the in-silico approach, the molecular docking study was performed to evaluate the possible antibacterial mechanism of W. somnifera phytochemicals such as Anaferine, Somniferine, Stigmasterol, Withaferin A, Withanolide- A, G, M, and Withanone by the inhibition of Penicillin- Binding Protein 4 (PBP4). Next, we utilized a bottom-up approach for the green synthesis of AgNPs- REWS, performed an in-detail phytochemical analysis, confirmed the AgNPs-REWS by SEM, UVvisible spectroscopy, XRD, FT-IR, and HPLC. Eventually, we examined their antibacterial activity. RESULTS: The result of molecular docking suggests that WS phytochemicals (Somniferine, Withaferin A, Withanolide A, Withanolide G, Withanolide M, and Withanone) possess the higher binding affinity toward the active site of PBP4 as compared to the Ampicillin (-6.39 kcal/mol) reference molecule. These phytochemicals predicted as potent inhibitors of PBP4. Next, as a proof-of-concept, AgNPs- REWS showed significant antibacterial effect as compared to crude, and control; against Xanthomonas and Ralstonia species. CONCLUSION: The in-silico and molecular docking analysis showed that active constituents of W. somnifera such as Somniferine, Withaferin A, Withanolide A, Withanolide G, Withanolide M, and Withanone possess inhibition potential for PBP4 and are responsible for the anti-bacterial property of W. somnifera extract. This study also establishes that AgNPs via the green synthesis with REWS showed enhanced antibacterial activity towards pathogenic bacteria.

Application of Artificial Intelligence in Pharmaceutical and Biomedical Studies.

BACKGROUND: Artificial intelligence (AI) is the way to model human intelligence to accomplish certain tasks without much intervention of human beings. The term AI was first used in 1956 with The Logic Theorist program, which was designed to simulate problem-solving ability of human beings. There have been a significant amount of research works using AI in order to determine the advantages and disadvantages of its applicabication and, future perspectives that impact different areas of society. Even the remarkable impact of AI can be transferred to the field of healthcare with its use in pharmaceutical and biomedical studies crucial for the socioeconomic development of the population in general within different studies, we can highlight those that have been conducted with the objective of treating diseases, such as cancer, neurodegenerative diseases, among others. In parallel, the long process of drug development also requires the application of AI to accelerate research in medical care. METHODS: This review is based on research material obtained from PubMed up to Jan 2020. The search terms include "artificial intelligence", "machine learning" in the context of research on pharmaceutical and biomedical applications. RESULTS: This study aimed to highlight the importance of AI in the biomedical research and also recent studies that support the use of AI to generate tools using patient data to improve outcomes. Other studies have demonstrated the use of AI to create prediction models to determine response to cancer treatment. CONCLUSION: The application of AI in the field of pharmaceutical and biomedical studies has been extensive, including cancer research, for diagnosis as well as prognosis of the disease state. It has become a tool for researchers in the management of complex data, ranging from obtaining complementary results to conventional statistical analyses. AI increases the precision in the estimation of treatment effect in cancer patients and determines prediction outcomes.