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1.
Front Aging Neurosci ; 13: 767493, 2021.
Article in English | MEDLINE | ID: mdl-34867295

ABSTRACT

Abnormal accumulation of misfolded proteins in the endoplasmic reticulum and their aggregation causes inflammation and endoplasmic reticulum stress. This promotes accumulation of toxic proteins in the body tissues especially brain leading to manifestation of neurodegenerative diseases. The studies suggest that deregulation of proteostasis, particularly aberrant unfolded protein response (UPR) signaling, may be a common morbific process in the development of neurodegeneration. Curcumin, the mixture of low molecular weight polyphenolic compounds from turmeric, Curcuma longa has shown promising response to prevents many diseases including current global severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and neurodegenerative disorders. The UPR which correlates positively with neurodegenerative disorders were found affected by curcumin. In this review, we examine the evidence from many model systems illustrating how curcumin interacts with UPR and slows down the development of various neurodegenerative disorders (ND), e.g., Alzheimer's and Parkinson's diseases. The recent global increase in ND patients indicates that researchers and practitioners will need to develop a new pharmacological drug or treatment to manage and cure these neurodegenerative diseases.

2.
Front Immunol ; 12: 819469, 2021.
Article in English | MEDLINE | ID: mdl-35095923

ABSTRACT

In malaria, anti-parasite immune response of the host may lead to dysregulated inflammation causing severe neuropathology arising from extensive damage to the Blood Brain Barrier (BBB). Use of anti-malarial drugs alone can control parasitemia and reduce inflammation but it cannot reduce pathology if chronic inflammation has already set in. In the present study, we have tested the efficacy of a new oral artemsinin based combination therapy (ACT) regimen using a combination of anti-malarial compounds like nanoartemisinin and nanoallylated-chalcone9 [{1-(4-Chlorophenyl)-3-[3-methoxy-4-(prop-2-en-1-yloxy) phenyl]-prop-2-en-1-one}]given together with anti-inflammatory-cum- anti-malarial compounds like nanoandrographolide and nanocurcumin to C57BL/6 mice infected with P. berghei ANKA. Untreated infected mice developed Experimental Cerebral Malaria (ECM) and died between 10 to 12 days after infection from severe BBB damage. We observed that oral treatments with nanoartemisinin or nano allylated chalcone 9 or nanoandrographolide alone, for 4 days after the onset of ECM, delayed the development of severe neurolopathology but could not prevent it. Nanocurcumin treatment for 4 days on the other hand, prevented damage to the BBB but the mice died because of hyperparasitemia. A single time oral administration of our ACT controlled blood parasitemia and prevented damage to the BBB, but recrudescence occurred due to persistence of parasites in the spleen. However the recrudescent parasites failed to induce ECM and BBB damage, leading to prolonged survival of the animals. A second time treatment at the start of recrudescence led to complete parasite clearance and survival of mice without pathology or parasitemia for 90 days. FACS analysis of spleen cells and gene expression profile in brain and spleen as well as quantitation of serum cytokine by ELISA showed that P. berghei ANKA infection in C57Bl/6 mice leads to a Th1-skewed immune response that result in severe inflammation and early death from ECM. Oral treatment with our ACT prevented a heightened pro-inflammatory response by modulating the Th1, Th2 and Treg immune responses and prevented ECM and death.


Subject(s)
Antimalarials/administration & dosage , Immunologic Factors/administration & dosage , Malaria/drug therapy , Malaria/parasitology , Nanoparticles , Plasmodium berghei/drug effects , Theranostic Nanomedicine , Animals , Blood-Brain Barrier , Brain/drug effects , Brain/immunology , Brain/parasitology , Cytokines/metabolism , Disease Models, Animal , Drug Therapy, Combination , Female , Malaria/immunology , Mice , Spleen/drug effects , Spleen/immunology , Spleen/parasitology
3.
Int J Mol Cell Med ; 10(3): 163-181, 2021.
Article in English | MEDLINE | ID: mdl-35178355

ABSTRACT

Malignant brain tumors proliferate aggressively and have a debilitating outcome. Surgery followed by chemo-radiotherapy has been the standard procedure of care since 2005 but issues of therapeutic toxicity and relapse still remain unaddressed. Repurposing of drugs to develop novel combinations that can augment existing treatment regimens for brain tumors is the need of the hour. Herein, we discuss studies documenting the use of curcumin as an adjuvant to conventional and alternative therapies for brain tumors. Comprehensive analysis of data suggests that curcumin together with available therapies can generate a synergistic action achieved through multiple molecular targeting, which results in simultaneous inhibition of tumor growth, and reduced treatment-induced toxicity as well as resistance. The review also highlights approaches to increase bioavailability and bioaccumulation of drugs when co-delivered with curcumin using nano-cargos. Despite substantial preclinical work on radio-chemo sensitizing effects of curcumin, to date, there is only a single clinical report on brain tumors. Based on available lab evidence, it is proposed that antibody-conjugated nano-curcumin in combination with sub-toxic doses of conventional or repurposed therapeutics should be designed and tested in clinical studies. This will increase tumor targeting, the bioavailability of the drug combination, reduce therapy resistance, and tumor recurrence through modulation of aberrant signaling cascades; thus improving clinical outcomes in brain malignancies.

4.
Clocks Sleep ; 2(4): 442-465, 2020 Oct 23.
Article in English | MEDLINE | ID: mdl-33114225

ABSTRACT

Sleep is essential for the survival of most living beings. Numerous researchers have identified a series of genes that are thought to regulate "sleep-state" or the "deprived state". As sleep has a significant effect on physiology, we believe that lack of total sleep, or particularly rapid eye movement (REM) sleep, for a prolonged period would have a profound impact on various body tissues. Therefore, using the microarray method, we sought to determine which genes and processes are affected in the brain and liver of rats following nine days of REM sleep deprivation. Our findings showed that REM sleep deprivation affected a total of 652 genes in the brain and 426 genes in the liver. Only 23 genes were affected commonly, 10 oppositely, and 13 similarly across brain and liver tissue. Our results suggest that nine-day REM sleep deprivation differentially affects genes and processes in the brain and liver of rats.

5.
Infect Immun ; 87(11)2019 11.
Article in English | MEDLINE | ID: mdl-31481412

ABSTRACT

Tuberculosis (TB) is one of the deadliest diseases, causing ∼2 million deaths annually worldwide. Mycobacterium bovis bacillus Calmette-Guérin (BCG), the only TB vaccine in common use, is effective against disseminated and meningeal TB in young children but is not effective against adult pulmonary TB. T helper 1 (Th1) cells producing interferon gamma (IFN-γ) and Th17 cells producing interleukin-17 (IL-17) play key roles in host protection against TB, whereas Th2 cells producing IL-4 and regulatory T cells (Tregs) facilitate TB disease progression by inhibiting protective Th1 and Th17 responses. Furthermore, the longevity of vaccine efficacy critically depends on the magnitude of long-lasting central memory T (TCM) cell responses. Hence, immunomodulators that promote TCM responses of the Th1 and Th17 cell lineages may improve BCG vaccine efficacy. Here, we show that curcumin nanoparticles enhance various antigen-presenting cell (APC) functions, including autophagy, costimulatory activity, and the production of inflammatory cytokines and other mediators. We further show that curcumin nanoparticles enhance the capacity of BCG to induce TCM cells of the Th1 and Th17 lineages, which augments host protection against TB infection. Thus, curcumin nanoparticles hold promise for enhancing the efficacy of TB vaccines.


Subject(s)
BCG Vaccine/immunology , Curcumin/pharmacology , Nanoparticles/administration & dosage , Tuberculosis/prevention & control , Adjuvants, Immunologic , Animals , Curcumin/administration & dosage , Female , Immunization , Mice , Mice, Inbred C57BL , Mycobacterium tuberculosis , Nanoparticles/chemistry
6.
Sci Rep ; 9(1): 4073, 2019 03 11.
Article in English | MEDLINE | ID: mdl-30858542

ABSTRACT

Vasculogenesis and angiogenesis are process of formation of blood vessels. Blood vessels are evolved to distribute nutrients and oxygen to distant organs. These vessels are crucial for growth and repair of wounded tissue. During tumor condition there occurs imbalance in the growth of blood vessels which leads to neo-angiogenesis. Neo-angiogenesis is major perpetrator behind the establishment of tumor. Tumor cells secrete pro-angiogenic factor VEGFA which binds to VEGFR2 present over surface of endothelial cells and triggers formation of new blood vessels. To inhibit tumor-angiogenesis, a physiologically-safe small molecule inhibitor was screened which can potentially interact with kinase domain of VEGFR2 and inhibit its activity. Molecular-docking module and biochemical analysis identified andrographolide as one of the best docking molecules that binds to ATP-binding pocket of VEGFR2 and inhibits its kinase activity. Thus, for a more radical approach towards safe VEGFR2 inhibitor, andrographolide was repurposed to inhibit tumor-angiogenesis and reduce tumor burden.


Subject(s)
Diterpenes/pharmacology , Neoplasms/drug therapy , Neovascularization, Pathologic/drug therapy , Vascular Endothelial Growth Factor A/genetics , Vascular Endothelial Growth Factor Receptor-2/genetics , Adenosine Triphosphate/chemistry , Adenosine Triphosphate/genetics , Andrographis paniculata , Angiogenesis Inhibitors/chemistry , Angiogenesis Inhibitors/pharmacology , Blood Vessels/drug effects , Blood Vessels/metabolism , Carrier Proteins/drug effects , Cell Movement/drug effects , Cell Proliferation/drug effects , Cell Survival/drug effects , Collagen/pharmacology , Diterpenes/chemistry , Drug Combinations , Human Umbilical Vein Endothelial Cells/drug effects , Humans , Laminin/pharmacology , Neoplasms/genetics , Neoplasms/pathology , Neovascularization, Pathologic/genetics , Neovascularization, Pathologic/pathology , Plant Extracts/chemistry , Protein Conformation/drug effects , Proteoglycans/pharmacology , Signal Transduction/drug effects , Vascular Endothelial Growth Factor A/chemistry , Vascular Endothelial Growth Factor Receptor-2/chemistry
7.
Sleep Sci ; 11(4): 245-253, 2018.
Article in English | MEDLINE | ID: mdl-30746042

ABSTRACT

BACKGROUND: Rapid Eye Movement sleep deprivation (REMSD) of rats causes inflammation of the liver and apoptotic cell death of neurons and hepatocytes. Studies also suggest that REM sleep deprivation can cause muscle as well as cardiac injury and neurodegenerative diseases. OBJECTIVE AND METHODS: The aim of this research was to determine whether REM sleep deprivation of rats would increase the levels of reactive oxygen species (ROS) in the hepatocytes and create oxidative stress in them. We selectively deprived the rats for REM sleep using the standard flower pot method. RESULTS: We observed that when rats were subjected to REM sleep deprivation, the levels of ROS in their hepatocytes increased ~184.33% compared to large platform control (LPC) group by day 9 of deprivation, but it returned towards normal level (~49.27%) after recovery sleep for 5 days. Nitric oxide synthase (iNOS) gene expression and protein levels as determined by real-time PCR and western blot analysis respectively were found to be elevated in hepatocytes of REM sleep deprived rats as compared to the LPC group. The level of nitric oxide (NO) in the hepatocytes of REMSD rats also increased by ~404.40% as compared to the LPC group but sleep recovery for 5 days normalized the effect (~135.35% compared to LPC group). We used a large platform control group as a reference group to compare with the REM sleep deprived group as the effect on the hepatocytes of both LPC group and cage control groups were not significantly different. DISCUSSION: We have analyzed the oxidative stress generated in the hepatocytes of rats due to REM sleep deprivation and further consequences of it. REMS deprivation not only increased the levels of ROS in the hepatocytes but also induced iNOS and NO in them. REM sleep deprived hepatocytes became more susceptible to oxidative stresses on further exposures. Furthermore, our study has great pathological and physiological.

8.
Front Immunol ; 8: 739, 2017.
Article in English | MEDLINE | ID: mdl-28713372

ABSTRACT

Curcumin, the bioactive component of turmeric also known as "Indian Yellow Gold," exhibits therapeutic efficacy against several chronic inflammatory and infectious diseases. Even though considered as a wonder drug pertaining to a myriad of reported benefits, the translational potential of curcumin is limited by its low systemic bioavailability due to its poor intestinal absorption, rapid metabolism, and rapid systemic elimination. Therefore, the translational potential of this compound is specifically challenged by bioavailability issues, and several laboratories are making efforts to improve its bioavailability. We developed a simple one-step process to generate curcumin nanoparticles of ~200 nm in size, which yielded a fivefold enhanced bioavailability in mice over regular curcumin. Curcumin nanoparticles drastically reduced hepatotoxicity induced by antitubercular antibiotics during treatment in mice. Most interestingly, co-treatment of nanoparticle-formulated curcumin along with antitubercular antibiotics dramatically reduced the risk for disease reactivation and reinfection, which is the major shortfall of current antibiotic treatment adopted by Directly Observed Treatment Short-course. Furthermore, nanoparticle-formulated curcumin significantly reduced the time needed for antibiotic therapy to obtain sterile immunity, thereby reducing the possibility of generating drug-resistant variants of the organisms. Therefore, adjunct therapy of nano-formulated curcumin with enhanced bioavailability may be beneficial to treatment of tuberculosis and possibly other diseases.

9.
Nanotechnology ; 28(16): 165101, 2017 Apr 21.
Article in English | MEDLINE | ID: mdl-28206982

ABSTRACT

The activation of cell-mediated and humoral immune responses to Mycobacterium tuberculosis (Mtb) is critical for protection against the pathogen and nanoparticle-mediated delivery of antigens is a more potent way to induce different immune responses. Herein, we show that mice immunized with Mtb lipid-bound chitosan nanoparticles (NPs) induce secretion of prominent type-1 T-helper (Th-1) and type-2 T-helper (Th-2) cytokines in lymph node and spleen cells, and also induces significantly higher levels of IgG, IgG1, IgG2 and IgM in comparison to control mice. Furthermore, significantly enhanced γδ-T-cell activation was observed in lymph node cells isolated from mice immunized with Mtb lipid-coated chitosan NPs as compared to mice immunized with chitosan NPs alone or Mtb lipid liposomes. In comparison to CD8+ cells, significantly higher numbers of CD4+ cells were present in both the lymph node and spleen cells isolated from mice immunized with Mtb lipid-coated chitosan NPs. In conclusion, this study represents a promising new strategy for the efficient delivery of Mtb lipids using chitosan NPs to trigger an enhanced cell-mediated and antibody response against Mtb lipids.


Subject(s)
Cytokines/metabolism , Intraepithelial Lymphocytes/immunology , Lipids/administration & dosage , Mycobacterium tuberculosis/chemistry , Nanoparticles/administration & dosage , Animals , Biocompatible Materials , Chitosan/chemistry , Endocytosis/physiology , Female , Humans , Immunity, Humoral/immunology , Immunoglobulin G/blood , Immunoglobulin M/blood , Intraepithelial Lymphocytes/drug effects , Macrophages, Peritoneal/drug effects , Mice, Inbred BALB C , Nanoparticles/chemistry , Tuberculosis, Pulmonary/drug therapy , Tuberculosis, Pulmonary/immunology
10.
High Alt Med Biol ; 17(4): 342-352, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27626325

ABSTRACT

Nehra, Sarita, Varun Bhardwaj, Santosh Kar, and Deepika Saraswat. Chronic hypobaric hypoxia induces right ventricular hypertrophy and apoptosis in rats: therapeutic potential of nanocurcumin in improving adaptation. High Alt Med Biol. 17:342-352, 2016.-a sustained work load on the right heart on ascent to high altitudes promotes right ventricular hypertrophy (RVH), which eventually undergoes decompensation and promotes pathological damage. However, the exact set of events leading to damage remains unidentified. Curcumin is a natural antioxidant and antihypertrophic agent, but it has poor biostability. Nanotized curcumin (nanocurcumin) has emerged as a promising agent with improved biostability while retaining the therapeutic properties of curcumin. The present study aimed at analyzing the therapeutic properties of nanocurcumin in ameliorating cardiac damage due to chronic hypobaric hypoxia (HH)-induced RVH in comparison to curcumin. Sprague-Dawley rats exposed to HH (25,000 feet, effective oxygen fraction in air [FIO2] ∼0.08, temperature 28°C ± 1°C, relative humidity 55% ± 2% for 3, 7, 14, and 21 days) developed RVH with increased interstitial collagen content, Fulton's index, and cardiomyocyte cross-sectional area while upregulating atrial natriuretic peptide. Tissue damage due to apoptotic cell death was evident by cytochrome-c/caspase-3 activation and TUNEL assay. Concomitant modulation of cyclic guanosine monophosphate (cGMP)/cGK-1, calmodulin-dependent protein kinase II (CaMkinase II), and intracellular calcium levels with increased free radical-induced damage and lipid peroxidation further contributed to the right heart pathology. Nanocurcumin supplementation decreased HH-induced RVH and apoptosis while modulating cardiac cGMP/cGK-1 signaling, and maintaining CaMkinase II, intracellular calcium levels and redox status better than curcumin. Nanocurcumin-mediated antiapoptotic effects might have benefited residents and sojourners at high altitude in preventing hypoxic cardiac damage.


Subject(s)
Altitude Sickness/physiopathology , Antioxidants/pharmacology , Apoptosis/drug effects , Curcumin/pharmacology , Hypertrophy, Right Ventricular/drug therapy , Nanostructures/therapeutic use , Acclimatization/drug effects , Altitude Sickness/complications , Animals , Hypertrophy, Right Ventricular/etiology , Male , Oxidation-Reduction/drug effects , Rats , Rats, Sprague-Dawley , Signal Transduction/drug effects
11.
Innate Immun ; 21(5): 546-52, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25466232

ABSTRACT

Dysregulation of the cytokine network in severe malaria owing to variations in factors like parasite load, strains and host factors is well documented but the key cytokines that are dysregulated remain poorly elucidated. Longitudinal changes in cytokine levels in an individual with parasitemia and disease resolution is likely to identify the key cytokines. We have analyzed the mRNA expression of cytokines over a 7-d period in severe (SM) and uncomplicated (UM) Plasmodium falciparum malaria. We found up-regulated expression of TNF-α, IL-1ß, IFN-γ and TGF-ß in SM, with decreased expression of IL-10 on d 0. Further, we observed a negative correlation of IL-10 expression with parasitemia and pro-inflammatory cytokines, suggesting IL-10 to be the key cytokine in tilting the balance to an inflammatory response. Longitudinal analysis revealed that the key cytokines associated with disease were TNF-α, IL-1ß, IFN-γ, IL-12α, RANTES and TGF-ß, while TNF-α, IL-10 and TGF-ß discriminated between SM and UM. A higher neutrophil count in SM and its positive association with parasite density and IL-1ß and IL-8 provides support for neutrophils in inflammation in malaria. Our findings suggest subversion of anti-inflammatory response in SM by parasite factors towards an exaggerated pro-inflammatory response with involvement of neutrophils, the classical inflammatory cells.


Subject(s)
Inflammation/metabolism , Inflammation/pathology , Interleukin-10/biosynthesis , Malaria, Falciparum/pathology , Neutrophils/pathology , Adolescent , Adult , Anemia/etiology , Animals , Child , Cytokines/metabolism , Female , Forkhead Transcription Factors/biosynthesis , Forkhead Transcription Factors/genetics , Humans , Leukocyte Count , Malaria, Falciparum/blood , Male , Middle Aged , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Young Adult
12.
Immunology ; 144(4): 561-73, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25284464

ABSTRACT

Tumour progression is associated with immune-suppressive conditions that facilitate the escape of tumour cells from the regimen of immune cells, subsequently paralysing the host defence mechanisms. Induction of CD4(+)  CD25(+)  FoxP3(+) T regulatory (Treg) cells has been implicated in the tumour immune escape mechanism, although the novel anti-cancer treatment strategies targeting Treg cells remain unknown. The focus of this study is to define the interaction between tumour and immune system, i.e. how immune tolerance starts and gradually leads to the induction of adaptive Treg cells in the tumour microenvironment. Our study identified hyperactivated mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) -signalling as a potential target for reversing Treg cell augmentation in breast cancer patients. In more mechanistic detail, pharmacological inhibitors of MEK/ERK signalling inhibited transforming growth factor-ß (TGF-ß) production in tumour cells that essentially blocked TGF-ß-SMAD3/SMAD4-mediated induction of CD25/interleukin-2 receptor α on CD4(+) T-cell surface. As a result high-affinity binding of interleukin-2 on those cells was prohibited, causing lack of Janus kinase 1 (JAK1)/JAK3-mediated signal transducer and activator of transcription 3 (STAT3)/STAT5 activation required for FoxP3 expression. Finally, for a more radical approach towards a safe MEK inhibitor, we validate the potential of multi-kinase inhibitor curcumin, especially the nano-curcumin made out of pure curcumin with greater bioavailability; in repealing tumour-shed TGF-ß-induced Treg cell augmentation.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Breast Neoplasms/drug therapy , Curcumin/pharmacology , MAP Kinase Kinase Kinases/antagonists & inhibitors , Paracrine Communication/drug effects , Protein Kinase Inhibitors/pharmacology , T-Lymphocytes, Regulatory/drug effects , Transforming Growth Factor beta/metabolism , Tumor Microenvironment , Animals , Breast Neoplasms/enzymology , Breast Neoplasms/genetics , Breast Neoplasms/immunology , Breast Neoplasms/pathology , Case-Control Studies , Chemistry, Pharmaceutical , Coculture Techniques , Dose-Response Relationship, Drug , Female , Forkhead Transcription Factors/metabolism , Humans , Interleukin-2 Receptor alpha Subunit/metabolism , MAP Kinase Kinase Kinases/genetics , MAP Kinase Kinase Kinases/metabolism , Mice, Inbred BALB C , Nanoparticles , STAT Transcription Factors/metabolism , Signal Transduction/drug effects , Smad Proteins/metabolism , T-Lymphocytes, Regulatory/immunology , T-Lymphocytes, Regulatory/metabolism , Time Factors , Transforming Growth Factor beta/genetics , Tumor Cells, Cultured , Tumor Escape/drug effects
13.
J Biol Chem ; 289(6): 3555-70, 2014 Feb 07.
Article in English | MEDLINE | ID: mdl-24297177

ABSTRACT

L-Asparaginase-II from Escherichia coli (EcA) is a central component in the treatment of acute lymphoblastic leukemia (ALL). However, the therapeutic efficacy of EcA is limited due to immunogenicity and a short half-life in the patient. Here, we performed rational mutagenesis to obtain EcA variants with a potential to improve ALL treatment. Several variants, especially W66Y and Y176F, killed the ALL cells more efficiently than did wild-type EcA (WT-EcA), although nonleukemic peripheral blood monocytes were not affected. Several assays, including Western blotting, annexin-V/propidium iodide binding, comet, and micronuclei assays, showed that the reduction in viability of leukemic cells is due to the increase in caspase-3, cytochrome c release, poly(ADP-ribose) polymerase activation, down-regulation of anti-apoptotic protein Bcl-XL, an arrest of the cell cycle at the G0/G1 phase, and eventually apoptosis. Both W66Y and Y176F induced significantly more apoptosis in lymphocytes derived from ALL patients. In addition, Y176F and Y176S exhibited greatly decreased glutaminase activity, whereas K288S/Y176F, a variant mutated in one of the immunodominant epitopes, showed reduced antigenicity. Further in vivo immunogenicity studies in mice showed that K288S/Y176F was 10-fold less immunogenic as compared with WT-EcA. Moreover, sera obtained from WT-EcA immunized mice and ALL patients who were given asparaginase therapy for several weeks recognized the K288S/Y176F mutant significantly less than the WT-EcA. Further mechanistic studies revealed that W66Y, Y176F, and K288S/Y176F rapidly depleted asparagine and also down-regulated the transcription of asparagine synthetase as compared with WT-EcA. These highly desirable attributes of these variants could significantly advance asparaginase therapy of leukemia in the future.


Subject(s)
Antineoplastic Agents , Asparaginase , Epitopes, B-Lymphocyte , Escherichia coli Proteins , Mutation, Missense , Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy , Amino Acid Substitution , Animals , Antineoplastic Agents/immunology , Antineoplastic Agents/pharmacology , Asparaginase/genetics , Asparaginase/immunology , Asparaginase/pharmacology , Caspase 3/genetics , Caspase 3/immunology , Caspase 3/metabolism , Cell Line, Tumor , Cytochromes c/genetics , Cytochromes c/immunology , Cytochromes c/metabolism , Epitopes, B-Lymphocyte/genetics , Epitopes, B-Lymphocyte/immunology , Epitopes, B-Lymphocyte/pharmacology , Escherichia coli Proteins/genetics , Escherichia coli Proteins/immunology , Escherichia coli Proteins/pharmacology , Female , Humans , Male , Mice , Mice, Inbred BALB C , Mutagenesis , Precursor Cell Lymphoblastic Leukemia-Lymphoma/immunology , bcl-X Protein/genetics , bcl-X Protein/immunology , bcl-X Protein/metabolism
14.
Biochimie ; 94(6): 1300-8, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22386870

ABSTRACT

A non-toxic, direct-acting fibrinolytic serine protease (Bafibrinase) demonstrating thrombolytic and anticoagulant properties was purified from Bacillus sp. strain AS-S20-I. Bafibrinase was monomeric, with a molecular mass of 32.3 kDa. The peptide mass fingerprinting of Bafibrinase revealed only 8.3% sequence coverage, suggesting it was a novel fibrinolytic enzyme. However, two of the tryptic digested de novo peptide sequences of Bafibrinase demonstrated good similarity with endopeptidases possessing serine in their catalytic triad. Further, catalytic activity of Bafibrinase was inhibited by serine protease inhibitor reinforcing this is a subtilisin-like serine protease. The apparent K(m) and V(max) values of Bafibrinase towards fibrin were determined as 0.24 µM and 2.8 µmol/min, respectively. It showed a K(m) value of 0.139 mM towards a chromogenic substrate for plasmin (D-Val-Leu-Lys-p-Nitroanilide dihydrochloride) and optimum activity at physiological conditions (37 °C and pH 7.4). Based on the cleavage pattern of fibrin and fibrinogen, Bafibrinase may be classified as an α,ß-fibrinogenase. Bafibrinase could not degrade collagen and was non-cytotoxic to HT29 cells or mammalian erythrocytes. Further, Bafibrinase at a dose of 2 mg/kg was devoid of toxicity as well as hemorrhagic activity on BALB/c mouse model, supporting its suitability for the development of a better and safer thrombolytic drug. Bafibrinase was also superior to human plasmin in degrading in vitro thrombus. The in vivo anticoagulant nature of Bafibrinase is being explored for the treatment and prevention of thrombosis and other cardiovascular diseases.


Subject(s)
Anticoagulants/pharmacology , Bacterial Proteins/metabolism , Bacterial Proteins/pharmacology , Fibrinolytic Agents/pharmacology , Serine Endopeptidases/metabolism , Serine Proteases/metabolism , Serine Proteases/pharmacology , Animals , Bacillus/enzymology , Bacterial Proteins/isolation & purification , Fibrin/metabolism , Fibrinolysis/drug effects , Fibrinolytic Agents/metabolism , Humans , Mice , Serine Proteases/isolation & purification , Thrombolytic Therapy
15.
Biotechnol Adv ; 30(1): 310-20, 2012.
Article in English | MEDLINE | ID: mdl-21619927

ABSTRACT

Curcumin has been shown to have anti malarial activity, but poor bioavailability and chemical instability has hindered its development as a drug. We have bound curcumin to chitosan nanoparticles to improve its bioavailability and chemical stability. We found that curcumin bound to chitosan nanoparticles did not degrade that rapidly in comparison to free curcumin when such particles were incubated in mouse plasma in vitro at room temperature. The uptake of bound curcumin from chitosan nanoparticles by mouse RBC was much better than from free curcumin. Oral delivery of curcumin bound chitosan nanoparticles to normal mice showed that they can cross the mucosal barrier intact and confocal microscopy detected the nanoparticles in the blood. Curcumin loaded chitosan nanoparticles when delivered orally improved the bioavailability of curcumin in the plasma and RBC. While mice infected with a lethal strain of Plasmodium yoelii (N-67) died between 8 and 9 days post infection, feeding of chitosan nanoparticles alone made them to survive for five more days. Feeding 1mg of native curcumin to infected mice per day for seven days resulted in survival of one third of mice but under the same condition when 1mg of curcumin bound to chitosan nanoparticles was fed all the mice survived. Like chloroquine, curcumin inhibited parasite lysate induced heme polymerization in vitro in a dose dependent manner and curcumin had a lower IC(50) value than chloroquine. We believe that binding of curcumin to chitosan nanoparticles increases its chemical stability and enhances its bioavailability when fed to mice. In vitro data suggest that it can inhibit hemozoin synthesis which is lethal for the parasite.


Subject(s)
Antimalarials/administration & dosage , Chitosan/chemistry , Curcumin/administration & dosage , Malaria/drug therapy , Nanoparticles/chemistry , Plasmodium yoelii/drug effects , Administration, Oral , Animals , Antimalarials/chemistry , Biological Availability , Curcumin/chemistry , Dose-Response Relationship, Drug , Drug Stability , Erythrocytes/drug effects , Female , Heme/metabolism , Inhibitory Concentration 50 , Mice
16.
Biochem Biophys Res Commun ; 410(2): 242-6, 2011 Jul 01.
Article in English | MEDLINE | ID: mdl-21651899

ABSTRACT

REM sleep is essential for maintenance of body physiology and its deprivation is fatal. We observed that the levels of ALT and AST enzymes and pro-inflammatory cytokines like IL-1 ß, IL-6 and IL-12 circulating in the blood of REM sleep deprived rats increased in proportion to the extent of sleep loss. But in contrast the levels of IFN-γ and a ∼200 kDa protein, identified by N-terminal sequencing to be alpha-1-inhibitor-3(A1I3), decreased significantly. Quantitative PCR analysis confirmed that REM sleep deprivation down regulates AII3 gene and up regulates IL1 ß, IL6 and their respective receptors gene expression in the liver initiating its inflammation.


Subject(s)
Acute-Phase Reaction/etiology , Liver/physiopathology , Sleep Deprivation/complications , Sleep Deprivation/physiopathology , Sleep, REM , Acute-Phase Proteins/biosynthesis , Acute-Phase Reaction/blood , Alanine Transaminase/blood , Animals , Aspartate Aminotransferases/blood , Cytokines/blood , Male , Rats , Rats, Wistar , Sleep Deprivation/blood
18.
Proteomics ; 9(17): 4192-208, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19722191

ABSTRACT

Wolbachia is an intracellular endosymbiont of Brugia malayi parasite whose presence is essential for the survival of the parasite. Treatment of B. malayi-infected jirds with tetracycline eliminates Wolbachia, which affects parasite survival and fitness. In the present study we have tried to identify parasite proteins that are affected when Wolbachia is targeted by tetracycline. For this Wolbachia depleted parasites (B. malayi) were obtained by tetracycline treatment of infected Mongolian jirds (Meriones unguiculatus) and their protein profile after 2-DE separation was compared with that of untreated parasites harboring Wolbachia. Approximately 100 protein spots could be visualized followed by CBB staining of 2-D gel and included for comparative analysis. Of these, 54 showed differential expressions, while two new protein spots emerged (of 90.3 and 64.4 kDa). These proteins were subjected to further analysis by MALDI-TOF for their identification using Brugia coding sequence database composed of both genomic and EST sequences. Our study unravels two crucial findings: (i) the parasite or Wolbachia proteins, which disappeared/down-regulated appear be essential for parasite survival and may be used as drug targets and (ii) tetracycline treatment interferes with the regulatory machinery vital for parasites cellular integrity and defense and thus could possibly be a molecular mechanism for the killing of filarial parasite. This is the first proteomic study substantiating the wolbachial genome integrity with its nematode host and providing functional genomic data of human lymphatic filarial parasite B. malayi.


Subject(s)
Brugia malayi/metabolism , Brugia malayi/microbiology , Helminth Proteins/metabolism , Parasites/metabolism , Parasites/microbiology , Tetracycline/pharmacology , Wolbachia/drug effects , Animals , Antibodies, Helminth , Brugia malayi/drug effects , Brugia malayi/immunology , Electrophoresis, Gel, Two-Dimensional , Electrophoresis, Polyacrylamide Gel , Immunoblotting , Parasites/drug effects , Polymerase Chain Reaction , Proteome/analysis , Reproducibility of Results
19.
Biochem Biophys Res Commun ; 364(3): 608-13, 2007 Dec 21.
Article in English | MEDLINE | ID: mdl-17963728

ABSTRACT

Ribonuclease S peptide and S protein offer a unique complementation system to understand the finer features of molecular recognition. In the present study the S peptide (1-16), and its retro and retro-inverso analogs have been analyzed for their structural and biological attributes. RPHPLC, CD, and NMR analyses have revealed that the physicochemical and conformational properties of the S peptide are distinct from those of its retro and retro-inverso analogs. On the functional side, while the S peptide complemented the S protein to give RNase activity, was recognized by anti-S peptide antibodies and induced T cell proliferation, neither the retro nor the retro-inverso S peptides could do so.


Subject(s)
Peptide Fragments/chemistry , Peptide Fragments/immunology , Ribonucleases/chemistry , Ribonucleases/immunology , T-Lymphocytes/immunology , Amino Acid Sequence , Binding Sites , Peptide Fragments/ultrastructure , Protein Binding , Ribonucleases/ultrastructure , Structure-Activity Relationship
20.
J Clin Immunol ; 27(4): 397-408, 2007 Jul.
Article in English | MEDLINE | ID: mdl-17372703

ABSTRACT

To characterize putatively protective immune response in bancroftian filariasis, Th1/Th2 cytokine profile induced in peripheral blood mononuclear cells (PBMCs) of endemic normal (EN) and asymptomatic microfilaremic (ASM) individuals were studied using different molecular weight fractions of Brugia malayi adult soluble antigens (BmA), which are differentially recognized by IgG4 antibodies present in their sera. Infection free and putatively immune individuals living in a filaria endemic area were identified and included in the present study as EN only after careful longitudinal follow up for three years. It was observed that the low molecular weight antigens present in Fr4 and Fr5 induced differential cytokine response; EN individuals showed a strong Th1 bias whereas ASM individuals showed a strong Th2 bias even though both the groups produced Th1 cytokines, albeit of different quantity, when a nonhelminthic antigen like H37Rv whole cell lysate was used. Since antigens present in Fr5 induced a highly polarized response, they should be examined for their diagnostic potential in lymphatic filariasis.


Subject(s)
Brugia malayi/immunology , Carrier State/immunology , Cytokines/metabolism , Filariasis/immunology , Immunoglobulin G/blood , Lymphocytes/immunology , Adolescent , Adult , Animals , Antigens, Helminth/blood , Antigens, Helminth/pharmacology , Cells, Cultured , Cytokines/immunology , Female , Filariasis/blood , Filariasis/epidemiology , Humans , Immunologic Tests , India/epidemiology , Lymphocytes/drug effects , Male , Microfilariae/immunology , Middle Aged , Molecular Weight , Solubility
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