Biomarkers in Alzheimer's disease.

Alzheimer's disease (AD) is among the most common neurodegenerative disorders. AD is characterized by deposition of neurofibrillary tangles and amyloid plaques, leading to associated secondary pathologies, progressive neurodegeneration, and eventually death. Currently used diagnostics are largely image-based, lack accuracy and do not detect early disease, ie, prior to onset of symptoms, thus limiting treatment options and outcomes. Although biomarkers such as amyloid-ß and tau protein in cerebrospinal fluid have gained much attention, these are generally limited to disease progression. Unfortunately, identification of biomarkers for early and accurate diagnosis remains a challenge. As such, body fluids such as sweat, serum, saliva, mucosa, tears, and urine are under investigation as alternative sources for biomarkers that can aid in early disease detection. This review focuses on biomarkers identified through proteomics in various biofluids and their potential for early and accurate diagnosis of AD.

Mechanistic insights and emerging therapeutic stratagems for Alzheimer's disease.

Alzheimer's disease (AD), a multi-factorial neurodegenerative disorder has affected over 30 million individuals globally and these numbers are expected to increase in the coming decades. Current therapeutic interventions are largely ineffective as they focus on a single target. Development of an effective drug therapy requires a deep understanding of the various factors influencing the onset and progression of the disease. Aging and genetic factors exert a major influence on the development of AD. Other factors like post-viral infections, iron overload, gut dysbiosis, and vascular dysfunction also exacerbate the onset and progression of AD. Further, post-translational modifications in tau, DRP1, CREB, and p65 proteins increase the disease severity through triggering mitochondrial dysfunction, synaptic loss, and differential interaction of amyloid beta with different receptors leading to impaired intracellular signalling. With advancements in neuroscience tools, new inter-relations that aggravate AD are being discovered including pre-existing diseases and exposure to other pathogens. Simultaneously, new therapeutic strategies involving modulation of gene expression through targeted delivery or modulation with light, harnessing the immune response to promote clearance of amyloid deposits, introduction of stem cells and extracellular vesicles to replace the destroyed neurons, exploring new therapeutic molecules from plant, marine and biological sources delivered in the free state or through nanoparticles and use of non-pharmacological interventions like music, transcranial stimulation and yoga. Polypharmacology approaches involving combination of therapeutic agents are also under active investigation for superior therapeutic outcomes. This review elaborates on various disease-causing factors, their underlying mechanisms, the inter-play between different disease-causing players, and emerging therapeutic options including those under clinical trials, for treatment of AD. The challenges involved in AD therapy and the way forward have also been discussed.

A network pharmacological approach for the identification of potential therapeutic targets of Brahmi Nei - a complex traditional Siddha formulation.

Brahmi Nei (BN), a traditional Indian polyherbal formulation has been described in classical texts for the treatment of anxiety and depression, as well as to fortify the immune system. The individual herbs of BN have been used for treatment of wide range of disorders including cognition, inflammation, skin ailments and cancer etc., This diverse basket of therapeutic activity suggests that BN may possess therapeutic benefits to other disorders. So, the present study aims to identify the potential therapeutic targets of BN using a network pharmacological approach to comprehend the multi target action of its multiple phytoconstituents. We have employed Randic Index for the first time to calculate the contribution score of module segregated targets towards diseases. Our results suggests that BN targets could also be effective in other diseases such as lysosomal storage disorders, respiratory disorders etc., apart from neurological disorders. The key targets with highest topological measures of Targets-(Pathway)-Targets network were identified as potential therapeutic targets of BN. And the top hit target PTGS2, a gene encoding for cyclooxygenase-2 was further evaluated using molecular docking, molecular dynamic simulation and in vitro studies. Our findings open up new therapeutic facets for BN that can be explored systematically in future.Communicated by Ramaswamy H. Sarma.

Methotrexate delivering microneedle patches for improved therapeutic efficacy in treatment of rheumatoid arthritis.

Arthritis is an inflammatory disorder that leads to degeneration and swelling in the joints thereby severely affecting mobility. Till date, a complete cure for this disorder remains elusive. Administration of disease modifying anti-rheumatic drugs has not proved effective owing to poor retention of drugs at the site of inflammation in the joints. In most cases, lack of adherence to the therapeutic regimen further aggravates the condition. Localized administration of the drugs through intra-articular injections is highly invasive and painful. A possible solution to overcome these issues will be to ensure sustained release of the anti-arthritic drug at the site of inflammation through a minimally invasive method. The present work focuses on the development of a microneedle patch for localized and minimally invasive delivery of methotrexate to arthritic joints in guinea pig model. The microneedle patch was found to elicit minimal immune response and ensured sustained release of the drug that was manifested through faster restoration of mobility and a distinct reduction in inflammatory and rheumatoid markers at the joints when compared to untreated and those treated through conventional hypodermic injections. Our results demonstrate the promise of microneedle-based platform for an effective arthritic therapy.

Self-assembled multifunctional nanotheranostics against circulating tumor clusters in metastatic breast cancer.

Circulating tumor clusters (CTC) disseminating from the primary tumor are responsible for secondary tumor formation where the conventional treatments such as chemotherapy and radiotherapy does not prevent the metastasis at locally advanced stage of breast cancer. In this study, a smart nanotheranostic system has been developed to track and eliminate the CTCs before it can colonize at a new site, which would reduce metastatic progression and increase the five-year survival rate of the breast cancer patients. Targeted multiresponsive (magnetic hyperthermia and pH) nanomicelles incorporated with NIR fluorescent superparamagnetic iron oxide nanoparticles were developed based on self-assembly for dual modal imaging and dual toxicity for spontaneous killing of CTCs in blood stream. A heterogenous tumor clusters model was developed to mimic the CTCs isolated from breast cancer patients. The nanotheranostic system was further evaluated for the targeting property, drug release kinetics, hyperthermia and cytotoxicity against developed CTC model in vitro. In vivo model in BALB/c mice equivalent to stage III and IV human metastatic breast cancer was developed to evaluate the biodistribution and therapeutic efficacy of micellar nanotheranostic system. Reduced CTCs in blood stream and low distant organ metastasis after treatment with the nanotheranostic system demonstrates its potential to capture and kill the CTCs that minimize the secondary tumor formation at distant sites.

Using Personal Activity Intelligence With Patients in a Clinic Setting: A Feasibility Study.

BACKGROUND: Personal Activity Intelligence (PAI) is a novel heart-rate-based metric used to assess cardiorespiratory fitness and quantify physical activity. OBJECTIVE: The aim of this study was to examine the feasibility, acceptability, and effectiveness of PAI with patients in a clinic setting. METHODS: Patients (n = 25) from 2 clinics underwent 12 weeks of heart-rate-monitored physical activity interfaced with aPAI Health phone app. We used a pre-post design with the Physical Activity Vital Sign and the International Physical Activity Questionnaire. Feasibility, acceptability, and PAI measures were used to evaluate the objectives. RESULTS: Twenty-two patients (88%) completed the study. There were significant improvements in International Physical Activity Questionnaire metabolic equivalent task minutes per week ( P = .046) and a decrease in sitting hours ( P = .0001). The Physical Activity Vital Sign activity increase in minutes per week was not significant ( P = .214). Patients achieved a mean PAI score of 116 ± 81.1 and 100 or greater 71% of the days. Most patients (81%) expressed satisfaction with PAI. CONCLUSIONS: Personal Activity Intelligence is feasible, acceptable, and effective when used with patients in a clinic setting.

Trends in Quantification of HbA1c Using Electrochemical and Point-of-Care Analyzers.

Glycated hemoglobin (HbA1c), one of the many variants of hemoglobin (Hb), serves as a standard biomarker of diabetes, as it assesses the long-term glycemic status of the individual for the previous 90-120 days. HbA1c levels in blood are stable and do not fluctuate when compared to the random blood glucose levels. The normal level of HbA1c is 4-6.0%, while concentrations > 6.5% denote diabetes. Conventionally, HbA1c is measured using techniques such as chromatography, spectroscopy, immunoassays, capillary electrophoresis, fluorometry, etc., that are time-consuming, expensive, and involve complex procedures and skilled personnel. These limitations have spurred development of sensors incorporating nanostructured materials that can aid in specific and accurate quantification of HbA1c. Various chemical and biological sensing elements with and without nanoparticle interfaces have been explored for HbA1c detection. Attempts are underway to improve the detection speed, increase accuracy, and reduce sample volumes and detection costs through different combinations of nanomaterials, interfaces, capture elements, and measurement techniques. This review elaborates on the recent advances in the realm of electrochemical detection for HbA1c detection. It also discusses the emerging trends and challenges in the fabrication of effective, accurate, and cost-effective point-of-care (PoC) devices for HbA1c and the potential way forward.

Emerging Trends in Nano-Driven Immunotherapy for Treatment of Cancer.

Despite advancements in the development of anticancer medications and therapies, cancer still has the greatest fatality rate due to a dismal prognosis. Traditional cancer therapies include chemotherapy, radiotherapy, and targeted therapy. The conventional treatments have a number of shortcomings, such as a lack of selectivity, non-specific cytotoxicity, suboptimal drug delivery to tumour locations, and multi-drug resistance, which results in a less potent/ineffective therapeutic outcome. Cancer immunotherapy is an emerging and promising strategy to elicit a pronounced immune response against cancer. Immunotherapy stimulates the immune system with cancer-specific antigens or immune checkpoint inhibitors to overcome the immune suppressive tumour microenvironment and kill the cancer cells. However, delivery of the antigen or immune checkpoint inhibitors and activation of the immune response need to circumvent the issues pertaining to short lifetimes and effect times, as well as adverse effects associated with off-targeting, suboptimal, or hyperactivation of the immune system. Additional challenges posed by the tumour suppressive microenvironment are less tumour immunogenicity and the inhibition of effector T cells. The evolution of nanotechnology in recent years has paved the way for improving treatment efficacy by facilitating site-specific and sustained delivery of the therapeutic moiety to elicit a robust immune response. The amenability of nanoparticles towards surface functionalization and tuneable physicochemical properties, size, shape, and surfaces charge have been successfully harnessed for immunotherapy, as well as combination therapy, against cancer. In this review, we have summarized the recent advancements made in choosing different nanomaterial combinations and their modifications made to enable their interaction with different molecular and cellular targets for efficient immunotherapy. This review also highlights recent trends in immunotherapy strategies to be used independently, as well as in combination, for the destruction of cancer cells, as well as prevent metastasis and recurrence.

Novel Combination Treatment for Melanoma: FLASH Radiotherapy and Immunotherapy Delivered by a Radiopaque and Radiation Responsive Hydrogel.

Immunotherapies have become the standard treatment for melanoma. To further improve patient responses, combinations of immunotherapies and radiotherapy (RT) are being studied, since radiotherapies can potentially provide additional immune stimulation, in addition to direct antitumor effects. FLASH-RT is a novel, ultrahigh dose rate, radiation delivery approach, with the potential of at least equivalent tumor control efficacy and reduced damage to healthy tissue. However, the effects of combining FLASH-RT and immunotherapy have not been extensively studied in melanoma. Toll-like receptor (TLR) agonists, such as imiquimod (IMQ), are potent immunostimulatory agents, although their utility is limited due to poor solubility and systemic side effects. We therefore developed a novel combination therapy for melanoma consisting of IMQ delivered to the tumor via a radiopaque and radiation responsive hydrogel combined with FLASH-RT. We found that FLASH was able to effectively stimulate IMQ release from the hydrogel. In addition, we found that the combination of FLASH and released IMQ resulted in synergistic melanoma cell killing in vitro. The combination therapy reduced tumor growth compared to controls, enhanced survival, and resulted in remarkable enhancements in certain tumor cytokine levels. CT imaging allowed the hydrogel to be monitored in vivo. In addition, no adverse effects of the treatment were observed. Overall, this IMQ-gel and FLASH-RT combination may have potential as an improved treatment for melanoma and indicates that the interactions of FLASH-RT and TLR agonists merit further study.

Ultrashort Peptide-Based Hydrogel for the Healing of Critical Bone Defects in Rabbits.

The use of hydrogels as scaffolds for three-dimensional (3D) cell growth is an active area of research in tissue engineering. Herein, we report the self-assembly of an ultrashort peptide, a tetrapeptide, Asp-Leu-IIe-IIe, the shortest peptide sequence from a highly fibrillogenic protein TDP-43, into the hydrogel. The hydrogel was mechanically strong and highly stable, with storage modulus values in MPa ranges. The hydrogel supported the proliferation and successful differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) in its matrix as assessed by cell viability, calcium deposition, alkaline phosphatase (ALP) activity, and the expression of osteogenic marker gene studies. To check whether the hydrogel supports 3D growth and regeneration in in vivo conditions, a rabbit critical bone defect model was used. Micro-computed tomography (CT) and X-ray analysis demonstrated the formation of mineralized neobone in the defect areas, with significantly higher bone mineralization and relative bone densities in animals treated with the peptide hydrogel compared to nontreated and matrigel treatment groups. The ultrashort peptide-based hydrogel developed in this work holds great potential for its further development as tissue regeneration and/or engineering scaffolds.

Artificial Intelligence-Driven Diagnosis of Pancreatic Cancer.

Pancreatic cancer is among the most challenging forms of cancer to treat, owing to its late diagnosis and aggressive nature that reduces the survival rate drastically. Pancreatic cancer diagnosis has been primarily based on imaging, but the current state-of-the-art imaging provides a poor prognosis, thus limiting clinicians' treatment options. The advancement of a cancer diagnosis has been enhanced through the integration of artificial intelligence and imaging modalities to make better clinical decisions. In this review, we examine how AI models can improve the diagnosis of pancreatic cancer using different imaging modalities along with a discussion on the emerging trends in an AI-driven diagnosis, based on cytopathology and serological markers. Ethical concerns regarding the use of these tools have also been discussed.

Investigations of an organic-inorganic nanotheranostic hybrid for pancreatic cancer therapy using cancer-in-a-dish andin vivomodels.

The incidence of highly aggressive pancreatic cancer is increasing across the globe and is projected to increase to 18.6% by 2050. The mortality rate for this form of cancer is very high and the 5 y relative survival rate is only about 9%-10%. The 3D pancreatic cancer microenvironment exerts a major influence on the poor survival rate. A key factor is the prevention of the penetration of the chemotherapeutic drugs in the three-dimensional (3D) microenvironment leading to the development of chemoresistance which is a major contributor to the survival rates. Hence,in vitrostudies using 3D cultures represent a better approach to understand the effect of therapeutic formulations on the cancer cells when compared to conventional 2D cultures. In the present study, we have explored three different conditions for the development of a 3D pancreatic tumour spheroid model from MiaPaCa-2 and PanC1 cells cultured for 10 days using Matrigel matrix. This optimized spheroid model was employed to evaluate a multi-functional nanotheranostic system fabricated using chitosan nanoparticles co-encapsulated with the chemotherapeutic agent gemcitabine and gold-capped iron oxide nanoparticles for multimodal imaging. The effect of the single and multiple-dose regimens of the theranostic system on the viability of 3D spheroids formed from the two pancreatic cancer cell lines was studied. It was observed that the 3D tumour spheroids cultured for 10 days exhibited resistance towards free gemcitabine drug, unlike the 2D culture. The administration of the multifunctional nanotheranostic system on alternate days effectively reduced the cancer cell viability after five doses to about 20% when compared with other groups. The repeated doses of the nanotheranostic system were found to be more effective than the single dose. Cell line-based differences in internalization of the carrier was also reflected in their response to the nanocarrier with PanC1 showing better sensitivity to the treatment.In vivostudies revealed that the combination of gemcitabine and magnetic field induced hypothermia produced superior regression in cancer when compared with the chemotherapeutic agent alone by a combination of activating the pro-apoptotic pathway and heat-induced necrosis. Our results reveal that this multi-functional system holds promise to overcome the current challenges to treat pancreatic cancers.

Vaginosis: Advances in new therapeutic development and microbiome restoration.

Vaginosis is a condition experienced by most women at least once in their lifetime. This condition arises due to the imbalance in the microbiome of the vaginal ecosystem. Most of the pathogens of this disease are organisms which are commonly found in a normal healthy vagina. The vaginal microbiome is important as they act as a primary defence against secondary infections and Sexually transmitted diseases and infections (STDs and STIs). The vagina is mostly dominated by Lactobacillus along with other microbes including Gardnerella vaginalis, Atopobium vaginae., Prevotella spp., Mobiluncus spp., etc. Vaginal microbiome also includes Candida albicans and other species of the genus. The ratio in which these species are present varies from person to person and the dominant species decides the whether a vagina is "normal" or not. Lactobacillus dominated vagina is considered normal and if dominated by Gardnerella and such it is considered to be Bacterial vaginosis (BV) and similarly for Vulvovaginal Candidiasis (VVC). The microbiome also undergoes changes during menstrual cycles and menopausal stages. Due to the dynamic nature of this microbiome, it is tough to perfectly restore the balance. But several treatments are currently available with antibiotics like Clindamycin and derivatives of 5-nitroimidazole drugs like Metronidazole. The extensive use and the non-adherence to the treatment regimen has led to drug resistance through biofilm formation, efflux pumps, single nucleotide polymorphisms and resulting recurrent episode of vaginosis in women. Alternative medicines, preparations from plant sources, anti-microbial peptides and nano formulations are also being explored. Most of these medicines tend to focus on reducing the pathogen load rather than restoring the balance of the ecosystem. Vaginal microbiome transplant, an effort to restore the normalcy in the vaginal environment is becoming a popular treatment. In this review we discuss about the types of vaginosis, available treatments, challenges in treating the condition and the new drugs that are under investigation.

Emerging trends in nano-bioactive-mediated mitochondria-targeted therapeutic stratagems using polysaccharides, proteins and lipidic carriers.

The emergence of new lifestyle disorders and pharmaco-resistant variants of diseases has necessitated the search for effective therapeutic moieties and approaches that could overcome the limitations in the existing treatment modalities. In this context, bioactives such as flavonoids, polyphenols, tannins, terpenoids and alkaloids have demonstrated promise in therapy owing to their ability to scavenge free radicals and modulate the mitochondrial function as well as regulate metabolic pathways. However, their clinical applicability is low owing to their poor bioavailability and aqueous solubility. The encapsulation of bioactives in nanodimensional particles has overcome these limitations to a large extent while simultaneously conferring additional advantages of improved circulation time, enhanced cell uptake and target specific release. A wide range of nanocarriers derived from biopolymers such as polysaccharides, lipids and proteins, have been explored for encapsulation of different bioactives and have reported significant improvement of the bioavailability and therapeutic efficacy of the encapsulated cargo. However, incorporation of cell-specific and mitochondria-specific elements on the nanocarriers has been relatively less explored. This review summarizes some of the recent attempts to treat different disorders using bioactives encapsulated in biopolymer nanostructures and few instances of mitochondria-specific delivery.

Nanoparticle-based strategies to target HIV-infected cells.

Antiretroviral drugs employed for the treatment of human immunodeficiency virus (HIV) infections have remained largely ineffective due to their poor bioavailability, numerous adverse effects, modest uptake in infected cells, undesirable drug-drug interactions, the necessity for long-term drug therapy, and lack of access to tissues and reservoirs. Nanotechnology-based interventions could serve to overcome several of these disadvantages and thereby improve the therapeutic efficacy of antiretrovirals while reducing the morbidity and mortality due to the disease. However, attempts to use nanocarriers for the delivery of anti-retroviral drugs have started gaining momentum only in the past decade. This review explores in-depth the various nanocarriers that have been employed for the treatment of HIV infections highlighting their merits and possible demerits.

A Pyrazolo Imine-based Colorimetric and Turn-on Fluorescent Sensor Probe for Determination of Hg2+ Ion and its Application in Test Paper Strips.

In this work, we synthesized diethylamino substituted pyrazolo imine (3) fluorescent probe for recognition of Hg2+ ion.The sensor probe 3 can detect Hg2+ by colorimetric method, and there is a 10-fold enhancement in fluorescence response. When the fluorescent probe bound with Hg2+ ion, turn-on fluorescence was observed via the coordination. Probe 3 has an excellent selectivity toward Hg2+ in the CH3 CN/H2 O (8:2, v/v) solution with low limit of detection and high binding association constant of 551 parts per billion (ppb) and 6.6067 × 106 m-1 for 3+Hg2+ , respectively. Furthermore, the formation of 3+Hg2+ complex with 1:1 binding mode was evidenced by Job's plot, 1 H NMR spectroscopy and Mass analysis. In addition, probe 3 is a feasible option to detect Hg2+ in various sources of water samples. Bio-imaging experiments have demonstrated that probe 3 can be used to monitor Hg2+ in Escherichia coli bacterial cell. The sensor 3 was also used for paper strip application to detect Hg2+ ion.

Passive immunotherapy using chicken egg yolk antibody (IgY) against diarrheagenic E. coli: A systematic review and meta-analysis.

BACKGROUND: Animal diarrhea due to diarrheagenic Escherichia coli (E. coli) has been a major concern in the field of livestock farming leading to a severe loss of domesticated animals. This systematic review aims to analyze medical shreds of evidence available in the literature and to discover the effect of IgY in treatment and protection against E. coli diarrhea. METHODS AND RESULTS: Research reports that aimed to evaluate the effect of IgY against E. coli diarrhea were searched and collected from several databases (Science Direct, Springer link, Wiley, T&F). The collected studies were screened based on the inclusion criteria. 19 studies were identified and included in the meta-analysis. The pooled relative risk ratios were calculated for the studies and found to be statistically significant to support the therapeutic effect of IgY against E. coli diarrhea but the 95% confidence interval of a majority of studies includes a relative risk of 1. This variability between the effect of IgY in the overall estimate and individual studies accounts due to the presence of methodological heterogeneity. In addition, subgroup analysis revealed the grounds for heterogeneity. CONCLUSIONS: This systematic review and meta-analysis provide concrete evidence for the favorable effect of IgY as a prophylactic and therapeutic modality against E. coli diarrhea. Yet, more research pieces of evidence with standardized animal studies aimed to utilize IgY against E. coli are vital. Further studies and trials on human subjects could open new perspectives in the application IgY as a therapeutic agent.

Towards understanding the mechanism of action of a polyherbal formulation using a multi-pronged strategy.

Herein, we investigate the cognitive effects of a traditional polyherbal formulation, Brahmi Nei (BN) for its effect on cognitive health. Network pharmacological analysis of the bioactives reported in the phytoconstituents of BN was performed by retrieving information from various databases. The in-silico predictions were experimentally validated using in vitro and in vivo models through a combination of biochemical, behavioural and molecular studies. The network pharmacological analysis of the key molecules in BN revealed their ability to modulate molecular targets implicated in memory, cognition, neuronal survival, proliferation, regulation of cellular bioenergetics and oxidative stress. Behavioral studies performed on normal adult rats administered with BN showed a significant improvement in their cognitive performance. Microarray analysis of their brain tissues exhibited an up-regulation of genes involved in oxidative phosphorylation, learning, neuronal differentiation, extension, regeneration and survival while pro-inflammatory and pro-degenerative genes were down-regulated. The oxygen consumption rate in BN-treated hippocampal cells showed a significant improvement in the bioenergetic health index when compared to untreated cells due to the mitochondrial membrane fortifying effect and anti-inflammatory property of the BN constituents. The neuroregenerative potential of BN was manifested in increase in axonal length and neurite outgrowth. Western blots and 2D gel electrophoresis revealed a reduction in pro-apoptotic proteins while increasing Akt and cyclophilin proteins. Taken together, our data reveal that BN, although traditionally used to treat anxiolytic disorders can be explored as a nutraceutical to improve neuronal health as well as a therapeutic option to treat cognitive disorders.