Awanish Kumar
NIT RAIPUR, Biotechnology, Faculty Member
- Dr. Awanish Kumar received his Ph.D. degree in Molecular Parasitology from Jawaharlal Nehru University (JNU), New Del... moreDr. Awanish Kumar received his Ph.D. degree in Molecular Parasitology from Jawaharlal Nehru University (JNU), New Delhi, India and CSIR-Central Drug Research Institute, Lucknow, India. After doing Post-doctoral study in molecular biology, proteomics, and therapeutic aspect of infectious disease at McGill University Canada, he has joined National Institute of Technology, Raipur, India as Assistant Professor, where he is working on various functions and therapeutic areas. The major research area of Dr. Awanish Kumar is disease biology and drug development. His current work is focused on antimicrobial resistance, drug targeting, drug development, and drug delivery of antimicrobial agents. He has guided many Ph.D, MS, and B.Tech students and such supervision are going on.edit
- Supervisoredit
Amyloidogenic pathway in Alzheimer’s disease (AD) involves breakdown of APP byβ-secretase followed byγ-secretase and results in formation of amyloid beta plaque.β-secretase has been a promising target for developing novel anti-Alzheimer... more
Amyloidogenic pathway in Alzheimer’s disease (AD) involves breakdown of APP byβ-secretase followed byγ-secretase and results in formation of amyloid beta plaque.β-secretase has been a promising target for developing novel anti-Alzheimer drugs. To test different molecules for this purpose, test ligands like acylguanidine 7a, rosiglitazone, pioglitazone, and tartaric acid were docked against our target proteinβ-secretase enzyme retrieved from Protein Data Bank, considering MK-8931 (phase III trial, Merck) as the positive control. Docking revealed that, with respect to their free binding energy, acylguanidine 7a has the lowest binding energy followed by MK-8931 and pioglitazone and binds significantly toβ-secretase.In silicoADMET predictions revealed that except tartaric acid all other compounds had minimal toxic effects and had good absorption as well as solubility characteristics. These compounds may serve as potential lead compound for developing new anti-Alzheimer drug.
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The visceral leishmaniasis (VL) caused by Leishmania donovani parasite severely affects large populations in tropical and subtropical regions of the world. The arsenal of drugs available is limited and resistance is common in clinical... more
The visceral leishmaniasis (VL) caused by Leishmania donovani parasite severely affects large populations in tropical and subtropical regions of the world. The arsenal of drugs available is limited and resistance is common in clinical field isolates. Therefore vaccines could be important alternative for prevention against VL. Recently, some investigators advocated the protective efficacy of DNA vaccines, which induces the T-cell based immunity against VL. The vaccine antigens are selected as conserved in various Leishmania species and provide a viable strategy for DNA vaccine development. Our understanding for DNA vaccine development against VL is not enough and much technological advancement is required. Improved formulations and methods of delivery are required, which increase the uptake of DNA vaccine by cells; optimization of vaccine vectors/encoded antigens to augment and direct the host immune response in VL. Despite the many genes identified as vaccine candidates, the disappo...
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GSK-3 is a member of cellular kinases with diversified functions such as cellular differentiation, metabolic signaling, neuronal functions and apoptosis. It has been validated as an important therapeutic target in... more
GSK-3 is a member of cellular kinases with diversified functions such as cellular differentiation, metabolic signaling, neuronal functions and apoptosis. It has been validated as an important therapeutic target in Alzheimer's disease and type 2 diabetes. Few molecules targeting GSK-3 are currently in clinical trials. In this study, we have compared certain docking and computational ADME (Absorption, Distribution, Metabolism, Excretion) parameters of a few GSK-3 targeted ligands (Indirubin, Hymenialdisine, Meridianins, 6-bromoindirubin-3-oxime) against two control molecules (Tideglusib and LY-2090314) to derive and analyze the basic drug-like properties of the test compounds. Docking between the GSK-3 and various ligands was done using AutoDock while ADME parameters were derived from ADMET server PreADMET and admetSAR. Various docked images were retrieved from docking, indicating the docking sites in the target protein. Out of four compounds tested, 6-bromoindirubin-3-oxime (6-BIO) was found as the best docking and ADME parameters, followed by Hymenialdisine (HMD). The LigPlot interaction results show two residues Leu (188) and Thr (138) to be common at the interaction site. The LD50 of 6-BIO is better than one of the control ligands while very similar to the other. Some of the parameters were very similar to the control ligands, thus, making it a suitable candidate among the test ligands. From this in-silico study, we concluded that 6-BIO is a potent drug candidate which could be further tested in vitro and in vivo to establish a drug molecule. Since, 6-BIO is a chemically modified form of the basic molecule Indirubin, we can hypothesize that certain other modified indirubins could be tested as GSK-3 targeted ligands.
Research Interests: Thermodynamics, Molecular Dynamics Simulation, Computer Software, Humans, Molecular docking, and 12 moreHydrogen Bonding, Protein Secondary Structure Prediction, Oximes, Oral Hypoglycemic Agents, THEORETICAL AND COMPUTATIONAL CHEMISTRY, Protein Binding, Ligands, Indole Alkaloids, Protein Kinase Inhibitors, Static Electricity, Biochemistry and cell biology, and binding sites
Synergistic effect of mixing of extractants (tri-n-butylphosphate, TBP, tri-n-octylamine, TOA, and Aliquat 336, A336) was investigated in view of recovery of pyruvic acid from fermentation media. Extraction experiments were performed at... more
Synergistic effect of mixing of extractants (tri-n-butylphosphate, TBP, tri-n-octylamine, TOA, and Aliquat 336, A336) was investigated in view of recovery of pyruvic acid from fermentation media. Extraction experiments were performed at T=302K, from aqueous solution of pyruvic acid (0.025-0.2 kmol m−3) using single and mixed extractants in decanol. Parameters such as distribution coefficient, KD, equilibrium constant, Keq, and extraction efficiency, E% were used to analyze the results. Equilibrium constants, Keq were evaluated to be 15.376 m3·kmol−1, 9.242 m3·kmol−1 and 0.0778 m3·kmol−1 for (TBP+TOA), (A336+TOA) and (TBP+A336), respectively. No overloading (Z<0.5) was observed and only (1:1) complex was proposed. E% were in the order of (TBP+TOA) > (A336+TOA) > (TBP+A336).
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Water is the most important and vital molecule of our planet and covers 75 % of earth surface. But it is getting polluted due to high industrial growth. The heavy metals produced by industrial activities are recurrently added to it and... more
Water is the most important and vital molecule of our planet and covers 75 % of earth surface. But it is getting polluted due to high industrial growth. The heavy metals produced by industrial activities are recurrently added to it and considered as dangerous pollutants. Increasing concentration of toxic heavy metals (Pb 2? , Cd 2? , Hg 2? , Ni 2?) in water is a severe threat for human. Heavy metal contaminated water is highly carcinogenic and poisonous at even relatively low concentrations. When they discharged in water bodies, they dissolve in the water and are distributed in the food chain. Bacteria and fungi are efficient microbes that frequently transform heavy metals and remove toxicity. The application of bacteria and fungi may offer cost benefit in water treatment plants for heavy metal transformation and directly related to public health and environmental safety issues. The heavy metals transformation rate in water is also dependent on the enzymatic capability of microorganisms. By transforming toxic heavy metals microbes sustain aquatic and terrestrial life. Therefore the application of microbiological biomass for heavy metal transformation and removal from aquatic ecosystem is highly significant and striking. This paper reviews the microbial transformation of heavy metal, microbe metal interaction and different approaches for microbial heavy metal remediation from water bodies.
Ectopic kidney is a well-known congenital anomaly. Intrathoracic kidney, as a form of renal ectopia, is an extremely rare entity. A 62-year-old man presented to the department of radiodiagnosis for evaluation of urinary problems. On the... more
Ectopic kidney is a well-known congenital anomaly. Intrathoracic kidney, as a form of renal ectopia, is an extremely rare entity. A 62-year-old man presented to the department of radiodiagnosis for evaluation of urinary problems. On the basis of ultrasonography, he was diagnosed to be having intrathoracic kidney, which was confirmed by CT scan. We present the details of the patient and brief review of the relevant literature.
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ABSTRACT Diabetes has been considered as the most dreaded non-communicable disease consuming the mankind rapidly. WHO has predicted the number of diabetics to be approximately 366 millions by 2030. The disease is characterized by... more
ABSTRACT Diabetes has been considered as the most dreaded non-communicable disease consuming the mankind rapidly. WHO has predicted the number of diabetics to be approximately 366 millions by 2030. The disease is characterized by hyperglycemia and the basic symptoms are polyphagia, polydipsia and polyuria. The autoimmune type 1 diabetes represent almost 1% of the total diabetic population, the rest being that of type 2 diabetes (T2D). Type 2 diabetes has been linked to a variety of factors such as heredity, environmental factors, unhealthy eating habits, sedentary lifestyle, stress etc. The uncontrolled hyperglycemia has profound deleterious effects on almost all the organs and results in various cardiovascular disorders, retinopathy, neuropathy, and nephropathy. Recent studies have revealed an array of pulmonary dysfunctions related with T2D ranging from respiratory defects to tuberculosis. Diabetes also predisposes the person to hepatic dysfunctions like NAFLD and HCC and a range of infections at various sites which are difficult to manage. Post-surgical infections are of special interest for subjects with uncontrolled hyperglycemia prior to surgery. Scientists all over the world are revealing different pathways and associated therapies for diabetes in order to control a monster which is destroying almost whole body physiology.
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Ionic liquids (ILs) have found various applications in the field of biotechnology that involves protein extraction from the aqueous phase. However, the stability of biomolecules in ILs is still unpredictable. Therefore, this work aims to... more
Ionic liquids (ILs) have found various applications in the field of biotechnology that involves protein extraction from the aqueous phase. However, the stability of biomolecules in ILs is still unpredictable. Therefore, this work aims to understand the effect of ammonium-based ILs with a fixed (trifluoromethylsulfonyl)imide [NTf2](-) anion and variable ammonium cations such as butyltrimethylammonium (IL-1), ethyldimethylpropylammonium (IL-2), diethylmethyl(2-methoxyethyl)ammonium (IL-3) and methyl-trioctylammonium (IL-4) on the stability of lysozyme. The spectroscopic analysis (UV, fluorescence and circular dichroism (CD)) revealed the existence of native structure of lysozyme in the presence of ILs at 25°C. Evidently, the presence of α-helix structure in lysozyme was confirmed using CD spectroscopy. In contrary, the thermal stability of the protein gradually decreased with increase in the concentration of the ILs. This was due to the strong favorable interactions of the ILs with th...
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Hernia is a common problem of the modern world with its incidence more in developing countries. Inguinal hernia is the most common groin hernia repaired worldwide. With advancement in technology operative techniques of repair have also... more
Hernia is a common problem of the modern world with its incidence more in developing countries. Inguinal hernia is the most common groin hernia repaired worldwide. With advancement in technology operative techniques of repair have also evolved. A PubMed and COCHRANE database search was accomplished in this regard to establish the current status of laparoscopic inguinal hernia repair in view of recent published literature. Published literature support that laparoscopic hernia repair is best suited for recurrent and bilateral inguinal hernia although it may be offered for primary inguinal hernia if expertise is available.
World Health Organization (WHO) considers obesity as one of the fastest growing metabolic disorders other than diabetes. It is a complex interplay of lifestyle and associated genes. Obesity has been considered as a disease with multiple... more
World Health Organization (WHO) considers obesity as one of the fastest growing metabolic disorders other than diabetes. It is a complex interplay of lifestyle and associated genes. Obesity has been considered as a disease with multiple targets and very often compared in this sense with its sibling disease type 2 diabetes. The disease is pathology of the adipocytes and develops as a result of hypertrophy and hyperplasia of these cells, former being the major concern but its effects could be seen on various organs in the form of cardio-vascular disease, stroke, cancer, diabetes, sexual dysfunction, respiratory problems and many more. An increase in the lipid content of the adipocytes changes the physiology of these cells towards more inflammatory phenotype. The array of molecules or adipokines secreted by these cells varies in concentration and type among healthy and obese subjects. On one hand where adiponectin concentration decreases, the resistin concentration increases resulting ...
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Amyloid-beta (Aβ) peptide mediates several neurodegenerative diseases. The 42 amino acid (Aβ1-42) is the predominant form of peptide found in the neuritic plaques and has been demonstrated to be neurotoxic in vivo and in vitro. The... more
Amyloid-beta (Aβ) peptide mediates several neurodegenerative diseases. The 42 amino acid (Aβ1-42) is the predominant form of peptide found in the neuritic plaques and has been demonstrated to be neurotoxic in vivo and in vitro. The availability of large quantities of Aβ peptide will help in several biochemical and biophysical studies that may help in exploring the aggregation mechanism and toxicity of Aβ peptide. We report a convenient and economical method to obtain such a peptide biologically. Synthetic oligonucleotides encoding Aβ1-42 were constructed and amplified through the polymerase cycling assembly (also known as assembly PCR), followed by the amplification PCR. Aβ1-42 gene was cloned into pET41a(+) vector for expression. Interestingly, the addition of 3% (v/v) ethanol to the culture medium resulted in the production of large amounts of soluble Aβ fusion protein. The Aβ fusion protein was subjected to a Ni-NTA affinity chromatography followed by enterokinase digestion, and the Aβ peptide was purified using glutathione Sepharose affinity chromatography. The peptide yield was ∼15mg/L culture, indicating the utility of this method for high-yield production of soluble Aβ peptide. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis and immunoblotting with anti-His antibody confirmed the identity of purified Aβ fusion protein and Aβ peptide. In addition, this method provides an advantage over the chemical synthesis and other conventional methods used for large-scale production of recombinant Aβ peptide.
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ABSTRACT
Isolation of the soluble exogenous antigens (SEAgs), its immune response study and proteome profiling is an essential prerequisite for understanding the molecular pathogenesis of Leishmania donovani. The immunostimulatory potential of L.... more
Isolation of the soluble exogenous antigens (SEAgs), its immune response study and proteome profiling is an essential prerequisite for understanding the molecular pathogenesis of Leishmania donovani. The immunostimulatory potential of L. donovani SEAgs, purified from culture of L. donovani clinical isolate was evaluated for their ability to induce cellular responses in treated/cured hamsters. SEAgs induced significant proliferative responses in lymphocytes (SI 5.6±2.3; P<0.01) isolated from cured hamster. In addition, significant NO production in response to SEAgs was also noticed in macrophages of hamsters, mouse and human cell lines (J774A-1 and THP1). Western blot analyses with antibodies against proteophosphoglycan (PPG; surface-expressed and secreted molecule) of L. donovani revealed that PPG molecules are also present in L. donovani SEAgs. Mass spectrometry (MS) based proteome analysis of 12 protein bands of SEAgs through MALDI-TOF/TOF endorsed the identification of some Th...
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To present a case report of trans-anal barotrauma by high-pressure compressed air jet as a dangerous practical joke, that is, playful insufflation of high-pressure air jet through the anal orifice resulting in sigmoid perforation. The... more
To present a case report of trans-anal barotrauma by high-pressure compressed air jet as a dangerous practical joke, that is, playful insufflation of high-pressure air jet through the anal orifice resulting in sigmoid perforation. The patient presented to emergency a day later with complaints of severe pain in the abdomen and abdominal distension following insufflation of high-pressure air jet through the anus. On examination, he had signs suggestive of perforation peritonitis and x-ray of the abdomen showed gas under the diaphragm. An emergency exploratory laparotomy was performed which revealed a 4-cm perforation in the sigmoid colon. Resection of the segment containing perforation along with the surrounding devitalised part was done with double-barrel colostomy. Reversal of colostomy was done after 8 weeks. Follow-up was uneventful.
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Bladder pheochromocytoma creates an extremely rare situation (0.06% of all bladder tumours). We came across a case with a complaint of intermittent episodes of haematuria. Cystoscopy revealed a solid, bluish submucosal growth with... more
Bladder pheochromocytoma creates an extremely rare situation (0.06% of all bladder tumours). We came across a case with a complaint of intermittent episodes of haematuria. Cystoscopy revealed a solid, bluish submucosal growth with increased vascularity. Immediately after taking biopsy, the patient developed a sharp headache, chest heaviness and marked rise in blood pressure. The procedure was abandoned. Later, serum and urinary markers for pheochromocytoma were within normal limits. The histopathology report came out to be pheochromocytoma of the bladder. Later, partial cystectomy was carried out under general anaesthesia and histopathology confirmed the same. Thus, a surgeon/urologist should have a high index of suspicion for bladder pheochromocytoma while dealing with such cases if he comes across the characteristic symptoms of sharp headache, hypertension, palpitation, sweating, fainting or blurring of vision immediately after voiding or during the cystoscopic manipulation of tum...
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We came across a case presenting with complaint of multiple fistulae present on back and lateral part of abdominal wall discharging faecal matter. Investigations revealed a fistulous tract arising from posterior wall of ascending colon.... more
We came across a case presenting with complaint of multiple fistulae present on back and lateral part of abdominal wall discharging faecal matter. Investigations revealed a fistulous tract arising from posterior wall of ascending colon. The patient did not respond to conservative management and therefore was subjected to exploratory laparotomy after thorough investigations. Peroperatively, an ileocaecal mass was found along with a fistulous tract which was arising from the posterior wall of ascending colon and communicating with the skin. A right haemicolectomy was performed followed by ileo-transverse anastomosis. Postoperative recovery was uneventful. The histopathology report came out to be adenocarcinoma of colon.
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Spontaneous enterocutaneous fistula is a rare entity. We encountered a case of spontaneous enterocutaneous fistula in the groin region due to femoral hernia. A 60-year-old man presented with spontaneous enterocutaneous fistula in the left... more
Spontaneous enterocutaneous fistula is a rare entity. We encountered a case of spontaneous enterocutaneous fistula in the groin region due to femoral hernia. A 60-year-old man presented with spontaneous enterocutaneous fistula in the left groin region without signs of peritonitis. He was kept on conservative treatment, but on third postadmission day, he developed a swelling in his right groin, which became firm and irreducible with signs of intestinal obstruction. On exploratory laparotomy, bilateral femoral hernias were noted with formation of enterocutaneous fistula on the left side. Reduction and repair of hernia was performed. In view of the rarity of this complication, this case is being reported here.
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Direct interactions between the anion and the catalytic amino acid residues lead to denaturation of CT.
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We encountered a patient who presented with neck swelling, difficulty in swallowing, voice change along with systemic features such as evening rise of temperature, chronic cough and weight loss. Ultrasonography of the thyroid gland... more
We encountered a patient who presented with neck swelling, difficulty in swallowing, voice change along with systemic features such as evening rise of temperature, chronic cough and weight loss. Ultrasonography of the thyroid gland revealed two cystic swellings. An ultrasound guided fine needle aspiration cytology was suggestive of tubercular abscess. The patient responded well to antigravity aspiration of the swellings and antitubercular treatment.
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A 35-year-old man developed priapism with the use of low-molecular-weight heparin and warfarin following repair of left brachial artery sustained after gunshot injury. Priapism progressed to penile gangrene despite decompression and... more
A 35-year-old man developed priapism with the use of low-molecular-weight heparin and warfarin following repair of left brachial artery sustained after gunshot injury. Priapism progressed to penile gangrene despite decompression and distal shunt procedure leading to total penectomy and perineal urethrostomy. We describe the mechanism of anticoagulant (heparin and warfarin)-induced penile gangrene and the possible methods to avert such a devastating complication.
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In the recent past years, a large number of proteins have been expressed in Escherichia coli with high productivity due to rapid development of genetic engineering technologies. There are many hosts used for the production of recombinant... more
In the recent past years, a large number of proteins have been expressed in Escherichia coli with high productivity due to rapid development of genetic engineering technologies. There are many hosts used for the production of recombinant protein but the preferred choice is E. coli due to its easier culture, short life cycle, well-known genetics, and easy genetic manipulation. We often face a problem in the expression of foreign genes in E. coli. Soluble recombinant protein is a prerequisite for structural, functional and biochemical studies of a protein. Researchers often face problems producing soluble recombinant proteins for over-expression, mainly the expression and solubility of heterologous proteins. There is no universal strategy to solve these problems but there are a few methods that can improve the level of expression, non-expression, or less expression of the gene of interest in E. coli. This review addresses these issues properly. Five levels of strategies can be used to increase the expression and solubility of over-expressed protein; (1) changing the vector, (2) changing the host, (3) changing the culture parameters of the recombinant host strain, (4) co-expression of other genes and (5) changing the gene sequences, which may help increase expression and the proper folding of desired protein. Here we present the resources available for the expression of a gene in E. coli to get a substantial amount of good quality recombinant protein. The resources include different strains of E. coli, different E. coli expression vectors, different physical and chemical agents and the co expression of chaperone interacting proteins. Perhaps it would be the solutions to such problems that will finally lead to the maturity of the application of recombinant proteins. The proposed solutions to such problems will finally lead to the maturity of the application of recombinant proteins.
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In recent years, ionic liquids (ILs) represent a new class of biocompatible co-solvents for biomolecules. In this work, we report the apparent transfer free energies... more
In recent years, ionic liquids (ILs) represent a new class of biocompatible co-solvents for biomolecules. In this work, we report the apparent transfer free energies (ΔG&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;tr) for six amino acids (AA) from water to aqueous solutions of six ammonium based ILs (diethylammonium acetate (DEAA), diethylammonium sulfate (DEAS), triethyl ammonium acetate (TEAA), triethylammonium sulfate (TEAS), triethylammonium dihydrogen phosphate (TEAP), and trimethylammonium acetate (TMAA)) through solubility measurements, as a function of IL concentration at 298.15 K under atmospheric pressure. Salting-out effect was found for AA in aqueous IL solutions with increasing IL concentrations. In addition, we observed positive values of ΔG&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;tr for AA from water to ILs, indicating that the interactions between ILs and AA are unfavorable. From the obtained results, we found that the selected ammonium based ILs act as stabilizers for the structure of AA.
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ABSTRACT Epsilon-poly-L-lysine (ɛ-PL) is a water-soluble, thermostable, biodegradable, and cationic homo-biopolymer with no toxicity to human health and the environment. It exhibits a characteristic peptide bond between the α-carboxyl... more
ABSTRACT Epsilon-poly-L-lysine (ɛ-PL) is a water-soluble, thermostable, biodegradable, and cationic homo-biopolymer with no toxicity to human health and the environment. It exhibits a characteristic peptide bond between the α-carboxyl groups and ɛ-amino groups of 25-35 L-lysine residues. It has a wide antimicrobial spectrum. Microorganisms such as bacteria and fungi do not easily develop resistance to it. ɛ-PL is polycationic in nature; therefore, it interferes with their cell membranes by ionic adsorption, leading to physiological damage to the microbial cell. ɛ-PL producers commonly possess a membrane-bound ɛ-PL-degrading aminopeptidase, which has high enzyme activity at pH above 4.0 and may play a role in self-protection. Because ɛ-PL is safe for human consumption and is biodegradable, therefore it has been used in many novel applications in the fields of food, medicine, environment, agriculture, and electronics in past decades. This review article addresses the occurrence, microbial synthesis, physiochemical properties, production enhancement, biodegradation, and potential applications of ɛ-PL.
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ABSTRACTPreviously, through a proteomic analysis, proliferating cell nuclear antigen (PCNA) was found to be overexpressed in the sodium antimony gluconate (SAG)-resistant clinical isolate compared to that in the SAG-sensitive clinical... more
ABSTRACTPreviously, through a proteomic analysis, proliferating cell nuclear antigen (PCNA) was found to be overexpressed in the sodium antimony gluconate (SAG)-resistant clinical isolate compared to that in the SAG-sensitive clinical isolate ofLeishmania donovani. The present study was designed to explore the potential role of the PCNA protein in SAG resistance inL. donovani. For this purpose, the protein was cloned, overexpressed, purified, and modeled. Western blot (WB) and real-time PCR (RT-PCR) analyses confirmed that PCNA was overexpressed by ≥3-fold in the log phase, stationary phase, and peanut agglutinin isolated procyclic and metacyclic stages of the promastigote form and by ∼5-fold in the amastigote form of the SAG-resistant isolate compared to that in the SAG-sensitive isolate.L. donovaniPCNA (LdPCNA) was overexpressed as a green fluorescent protein (GFP) fusion protein in a SAG-sensitive clinical isolate ofL. donovani, and modulation of the sensitivities of the transfec...
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ABSTRACT The early evolution of acute kidney injury (AKI) in humans is difficult to study noninvasively. We hypothesized that urine proteomics could provide insight into the early pathophysiology of human AKI. A prospective nested... more
ABSTRACT The early evolution of acute kidney injury (AKI) in humans is difficult to study noninvasively. We hypothesized that urine proteomics could provide insight into the early pathophysiology of human AKI. A prospective nested case-control study (n = 250) compared serial urinary proteomes of 22 patients with AKI and 22 patients without AKI before, during, and after cardiopulmonary bypass surgery. AKI was defined as a greater than 50% increase in serum creatinine level, and non-AKI, as less than 10% increase from baseline. Serum creatinine, urine protein-creatinine ratio, neutrophil gelatinase-associated lipocalin (NGAL), alpha1-microglobulin, interferon-inducible protein-10 (IP-10), monokine induced by interferon gamma (Mig), interferon-inducible T cell alpha chemoatractant (I-TAC), interleukin 6 (IL-6), IL-1beta, and IL-10. Urine protein profiling by means of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). SELDI-TOF-MS showed intraoperative tubular stress in both groups on arrival to the intensive care unit, evidenced by beta2-microglobulinuria. Non-AKI proteomes returned toward baseline postoperatively. In contrast, AKI proteomes showed a second phase of tubular injury/stress with the reappearance of beta2-microglobulin and multiple unidentified peaks (3 to 5 and 6 to 8 kDa) and the appearance of established tubular injury markers: urinary protein, alpha1-microglobulin, and NGAL. Furthermore, 2 novel peaks (2.43 and 2.78 kDa) were found to be dominant in postoperative non-AKI urine samples. The 2.78-kDa protein was identified as the active 25-amino acid form of hepcidin (hepcidin-25), a key regulator of iron homeostasis. Finally, an inflammatory component of reperfusion injury was evaluated by means of enzyme-linked immunosorbent assay analysis of candidate chemokines (IP-10, I-TAC, and Mig) and cytokines (IL-6, IL-1beta, and IL-10). Of these, IP-10 was upregulated in patients with versus without AKI postoperatively. This is an observational study. SELDI-TOF-MS is a semiquantitative technique. Evaluation of human AKI revealed early intraoperative tubular stress in all patients. A second phase of injury observed in patients with AKI may involve IP-10 recruitment of inflammatory cells. The enhancement of hepcidin-25 in patients without AKI may suggest a novel role for iron sequestration in modulating AKI.
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Prolonged diabetes ultimately leads to Diabetic Retinopathy (DR) which is one of the leading causes of preventable blindness in the world. Through advanced image analysis techniques are used for abnormalities detection in retina that... more
Prolonged diabetes ultimately leads to Diabetic Retinopathy (DR) which is one of the leading causes of preventable blindness in the world. Through advanced image analysis techniques are used for abnormalities detection in retina that define and correlate the severity of DR. A thorough study is done in this area in recent past years and on the basis of these studies we have developed a computer based prediction model that is used to determine the severity of DR. To identify severity DR, we have analyzed the human eye image. We have extracted some important features from human eye image i.e. Blood Artery, Optical disc, Exudates. Based on these image and data we have designed an automated system for the determination of DR severity. This automated DR severity assessment methods can be used to predict the clinical case and conditions when young clinicians would agree or disagree with their more experienced fellow members. The algorithms described in this study may be used in clinical pr...
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Helicobacter pylori are the well known human pathogen associated with gastric cancer and peptic ulcer. Pathogenesis is mainly due to the presence of 40kb cagPAI (cag Pathogenicity Island) region that encodes the type IV secretion system... more
Helicobacter pylori are the well known human pathogen associated with gastric cancer and peptic ulcer. Pathogenesis is mainly due to the presence of 40kb cagPAI (cag Pathogenicity Island) region that encodes the type IV secretion system (TFSS) consisting of a cytoplasmic part, a middle part/core complex (spans from inner membrane to outer membrane), and an outer membrane associated part. CagX and CagT are two important proteins of TFSS that have homology with virB9 and virB7 of Agrobacterium tumefaciens TFSS. In this study, we have shown that the CagX and CagT interact directly by using co-immunoprecipitation of endogenous CagX and CagT and MBP pull down assay. We further authenticate this observation using yeast two-hybrid assay and co-expression of both the protein coding gene in Escherichiacoli. We also observed that the C-terminal region of CagX is important for CagT interaction. We reconfirm that CagT depends on CagX for its stabilization. These observations could contribute in...
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Alzheimer's disease (AD) is a neurodegenerative disorder in which the death of brain cells causes memory loss and cognitive decline i.e. dementia. The disease starts with mild symptoms and gradually becomes severe. AD is one of the... more
Alzheimer's disease (AD) is a neurodegenerative disorder in which the death of brain cells causes memory loss and cognitive decline i.e. dementia. The disease starts with mild symptoms and gradually becomes severe. AD is one of the leading causes of mortality worldwide. Several different hallmarks of the disease have been reported such as deposits of β-amyloid around neurons, hyperphosphorylated tau protein, oxidative stress, dyshomeostasis of bio-metals, low levels of acetylcholine etc. AD is not simple to diagnose since there is no single diagnostic test for it. Pharmacotherapy for AD currently provides only symptomatic relief and mostly targets cognitive revival. Computational biology approaches have proved to be reliable tools for selection of novel targets and therapeutic ligands. Molecular docking is a key tool in computer-assisted drug design and development. Docking has been utilized to perform virtual screening on large libraries of compounds, and propose structural hypotheses of how the ligands bind with the target with lead optimization. Another potential application of docking is optimization stages of the drug-discovery cycle. This review summarizes the known drug targets of AD, in vivo active agents against AD, state-of-the-art docking studies done in AD and future prospects of the docking with particular emphasis on AD.
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Leishmania donovani, a causative organism of visceral leishmaniasis (VL), is responsible for high mortality throughout the world. Due to the unsatisfactory treatment measures and increasing drug resistance, there has been an urgent need... more
Leishmania donovani, a causative organism of visceral leishmaniasis (VL), is responsible for high mortality throughout the world. Due to the unsatisfactory treatment measures and increasing drug resistance, there has been an urgent need to develop novel drug/vaccine targets against VL. The aim of this study was to identify novel targets in soluble L. donovani (SLD) protein. SLD protein was isolated and resolved by two-dimensional gel electrophoresis and analyzed through MALDI-TOF/TOF-based mass spectrometry. Proteomic results identified several proteins as drug targets, Th1 stimulatory, novel, and hypothetical proteins which could have crucial biological functions in Leishmania pathogenesis.