Koushlesh Ranjan

Nanotherapeutics: An insight into healthcare and multi-dimensional applications in medical sector of the modern world

In recent years nanotechnology has revolutionized the healthcare strategies and envisioned to have a tremendous impact to offer better health facilities. In this context, medical nanotechnology involves design, fabrication, regulation, and application of therapeutic drugs and devices having a size in nano-range (1-100 nm). Owing to the revolutionary implications in drug delivery and gene therapy, nanotherapeutics has gained increasing research interest in the current medical sector of the modern world. The areas which anticipate benefits from nano-based drug delivery systems are cancer, diabetes, infectious diseases, neurodegenerative diseases, blood disorders and orthopedic problems. The development of nanotherapeutics with multi-functionalities has considerable potential to fill the lacunae existing in the present therapeutic domain. Nanomedicines in the field of cancer management have enhanced permeability and retention of drugs thereby effectively targeting the affected tissues. Polymeric conjugates of asparaginase, polymeric micelles of paclitaxel have been recmended for various types of cancer treatment .The advancement of nano therapeutics and diagnostics can provide the improved effectiveness of the drug with less or no toxicity concerns. Similarly, diagnostic imaging is having potential future applications with newer imaging elements at nano level. The newly emerging field of nanorobotics can provide new directions in the field of healthcare. In this article, an attempt has been made to highlight the novel nanotherapeutic potentialities of polymeric nanoparticles, nanoemulsion, solid lipid nanoparticle, nanostructured lipid carriers, dendrimers, nanocapsules and nanosponges based approaches. The useful applications of these nano-medicines in the field of cancer, nutrition, and health have been discussed in details. Regulatory and safety concerns along with the commercial status of nanosystems have also been presented. In summary, a successful translation of emerging nanotherapeutics into commercial products may lead to an expansion of biomedical science. Towards the end of the review, future perspectives of this important field have been introduced briefly.

Satellite Imaging and Surveillance of Infectious Diseases

The Geographical Information Systems (GIS) and satellite image data can provide useful information for detection and management of both human and animal diseases outbreaks. The satellite surveillance can be used for monitoring of several environmental variables such as temperature, precipitation, humidity, wind speed and direction etc that influences the activity of pathogens, vectors and their interactions with human and animal hosts. By statistical analysis of satellite surveillance data the models based on geographic and vegetation of a particular landscape providing conducive environment to pathogens, spatial and temporal factors determining the distribution of disease can be framed. The GIS data analysis may help in several aspects during outbreak such as identification and spread of diseases over time, population groups at risk, patterns of disease outbreaks, facility available to healthcare and program intervention planning and assessment in disease outbreak. The satellite surveillance have been used in study of several water and vector borne diseases such as diarrhoea, cholera, typhoid, leptospirosis, Rift Valley Fever, Foot and mouth disease, bluetongue, West Nile Virus disease, Japanese encephalitis etc. The remote sensing and GIS data analysis is proved as powerful tools for disease surveillance, predicting its outbreaks, and monitoring control programs.

Biomineralization Abilities of Cupriavidus Strain and Bacillus subtilis Strains In Vitro Isolated from Speleothems, Rani Cave, Chhattisgarh, India

ABSTRACT In vitro culture experiments using three bacterial strains CSJC1, CSJC2, and CSJC3 isolated from speleothems, Rani cave, Chhattisgarh, India, were studied to examine their biomineralization potential. These speleothems showed high microbial cell enumerations on nutrient agar and iron agar (9 × 104 CFU/g) followed by thiosulfate agar (7 × 104 CFU/g), and 60 diverse strains were isolated. The BLASTn sequence search of 16S rRNA sequences with the NCBI database to establish the identity of CSJC1, CSJC2, and CSJC3 strains yielded similarity scores of ≥99% with the respective organisms, and the strains were identified as CSJC1 – Bacillus sp., CSJC2 – Cupriavidus sp., CSJC3 – Bacillus sp. The phylogenetic analysis of CSJC2 strain suggests that it formed a separate major cluster with Cupriavidus sp. and Cupriavidus necator. The phylogenetic analysis of CSJC1 and CSJC3 strains revealed that it formed a major cluster with several strains of Bacillus sp. and Bacillus subtilis. The biominerals induced by Cupriavidus sp. CSJC2 strain imaged with an ultra high-resolution field emission scanning electron microscope (FE-SEM) were seen as calcified coccoid shells that transformed into calcified dumbbells. FE-SEM imaging of biominerals induced by B. subtilis CSJC1 and CSJC3 tested both on B4 media and sheep blood agar individually showed that the precipitates formed calcified dumbbells that were almost similar but not identical phenotypically, indicating that strain-specific morphologies and crystal formation is easier when Ca is present in the media. This is the first comprehensive report on the possible evidences about the role of Cupriavidus sp. in calcite precipitation isolated from speleothems in the Indian caves. These results allow us to postulate that the identified strains may have a role in the biogenic influences in mineral formations at Rani cave.

Nanodiagnostics: a new frontier for veterinary and medical sciences.

Infectious diseases are one of the greatest threats to animal and human population living in the developing world. These diseases have capacity to instigate in a small area and then open out very fast to the rest of the world and causing a heavy pandemic situation, for example; avian influenza pandemic. Such diseases infect large masses of population and may lead to loss of lives and also incur huge economic losses. Therefore, the best way to control these diseases is by diagnosing it at a very primary level and taking necessary precautionary measures so as to avoid the spread. Since last few years, the diagnostic approach has changed from tedious molecular biological techniques, to easy and rapid diagnostic techniques. Nanotechnology has extended the molecular diagnostics limit to nanoscale. These developed techniques do not require sophisticated laboratories and expert personnel, and hence are a cheap diagnostic approach. These assays can also be performed at the field level where the patient is present and get the results there itself. Hence, they are also called as pen side test or lab on chip diagnostic assays. The biological tests using nanotechnology become quicker, more flexible and more sensitive. These techniques have greatly influenced the diagnostic approach in the veterinary as well as medical field. Especially in the developing countries such as India, where the laboratory services are not Upendra Lambe 1 , Minakshi P 1, *, Basanti Brar 1 , Madhusudan Guray 1 , Ikbal 1 , Koushlesh Ranjan 2 , Nitish Bansal 1 , Sandip Kumar Khurana 3 and Manimegalai J 1

BATS: CARRIERS OF ZOONOTIC VIRAL AND EMERGING INFECTIOUS DISEASES

Bats are reported as reservoir host for several viruses, which cause significant illness in human and animals. Some of the bat transmitted zoonotic viral diseases such as Ebola, Hendra, Nipah and rabies may cause severe human casualties. They also harbor several other viruses such as MERS and SARS corona viruses, which may cause disease in human through direct spillover to human or through an intermediate host or vectors. Being reservoir hosts bats do not get affected by these viruses. This probably may happen due to the specificity of bat immune system, which reacts differently with viral pathogens in comparison to their other mammalian counterparts. Although bats are important reservoir hosts for several zoonotic viruses, very little information is available regarding host/virus relationships as only few experimental studies have been done on bat colonies, lack of expertise for study of bat immunology and antiviral responses and difficulty in conducting field work. However, with the advancement in epidemiology and molecular biology, these problems can be addressed, which will provide the insight into interactions of bats and zoonotic viruses. It may also clarify regarding virus persistence in nature and various associated risk factors which might facilitate viral transmission to animals and humans. Koushlesh Ranjan 1,* , Minakshi Prasad 2 and Gaya Prasad 3

Occurrence of Neglected Picobirna Virus in Buffalo Calves Associated with Diarrhoea

Picobirna viruses (PBVs) thought to be neglected viruses, are emerging as causative agents of viral gastrointestinal infections in human and other animal species with enteritis. In the present study, a total of 408 fecal samples of diarrheic buffalo calves of under age of 2 months were collected from different locations in Haryana. These samples were preliminarily screened for PBV by RNApolyacrylamide gel electrophoresis (RNA-PAGE). The RNA-PAGE positive samples were further confirmed by RT-PCR using RdRp gene specific primer pair PicoB25 (+) and PicoB43 (-) for amplification of genogroup I PBV that generated an amplicon of 201 bp. Out of 408 buffalo calves fecal samples, 52 (13%) samples were found positive for genogroup 1(GGI) PBV using RT-PCR. Genogroup II (GGII) specific primer generating 369 bp product size did not show amplification in any of the samples. The results suggested occurrence of PBV in Haryana State with predominance of GGI PBV. To the best of our knowledge, this is the first report of detection as well as occurrence of GGI picobirna virus among diarrheic buffalo calves in Haryana State.

Virus-Host Interactions: New Insights and Advances in Drug Development Against Viral Pathogens

BACKGROUND Viruses are the most devastating pathogens of almost all life forms including humans and animals. Viruses can replicate very fast and may affect any metabolic and physiological function of the host cell. Therefore, it has been a challenge to develop a universal and common treatment against viral pathogens, in contrast to bacterial pathogens. Virus-host interaction is a complex phenomenon and often is virus- and host cell-specific. Exciting new insights into the molecular pathogenesis and host-virus interactions have been gained over the past few decades. These advances have enabled researchers to design better antiviral drugs. METHODS The literature related to various aspects of virus-host interactions: new insights and advances in drug development was collected from several scientific research related databases such as Science Direct, Google Scholar, Scopus, PubMed, AGRICOLA, and Medline, etc. Total number of 319 research papers was used to compile the information regarding drug development against viral pathogens. RESULTS Clinical adequacy of antiviral drugs and their bioavailability are important parameters for effective treatment of viral infections. The problems associated with effective delivery of a drug in a safe and desired quantity have led to the search for (and design of) better drug delivery systems. In recent past, several new antiviral drugs have been developed, which have high therapeutic effectiveness against life-threatening viral diseases such as HIV, hepatitis B virus, herpes virus, dengue virus and influenza virus infections. The majority of recent advances in antiviral drug discovery were possible due to the developments in allied fields such as in vitro virus cultivation technology, molecular biology of viral-genome-encoded enzymes, complete-genome-sequence-based studies of viruses and identification of suitable targets for antiviral drugs in viral genomes. Recently, several novel drug delivery approaches including small interfering RNAs (siRNAs) have emerged to aid antiviral therapy. CONCLUSION The present review is aimed at providing an update on research and development efforts being made to create effective antiviral chemotherapeutic agents and approaches to their delivery to appropriate targeted cells or tissues.

Synthesis, characterization and anticancer activity of vincristine loaded folic acid-chitosan conjugated nanoparticles on NCI-H460 non-small cell lung cancer cell line

Abstract Vincristine is used to treat different type of cancers. But unwanted side effects limit their applications in medicine. To overcome these side effects by targeted drug delivery approach, we synthesized vincristine loaded folic acid-chitosan conjugated nanoparticles by ionic gelation method. The nanoparticles were characterized using Dynamic Light Scattering (DLS), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy. DLS confirmed small sized nanoparticles (<200 nm) while FTIR confirmed different functional groups associated with synthesized nanoparticles. SEM showed spherical shaped nanoparticles with smooth surface, whereas, TEM confirmed loading of vincristine in folic acid-chitosan conjugated nanoparticles. Anticancer activity of vincristine loaded folic acid-chitosan conjugated nanoparticles was checked by MTT assay on NCI-H460 cells followed by reactive oxygen species level, mitochondrial transmembrane potential and apoptotic morphological changes, which confirmed the cytotoxicity of nanoparticles. Erythrocyte aggregation assay confirmed non-toxicity of nanoparticles. Hence, these nanoparticles can be used to treat NCI-H460 cells.

First Report of Isolation of Bluetongue Virus 23 from Culicoides Peregrinus Vector from India

Bluetongue (BT) disease is caused by bluetongue virus (BTV) of Reoviridae family. BTV is transmitted by several species of biting midges (Culicoides sp.) to domestic and wild ruminants. BT is non-contagious but infectious disease. BTV isolate was isolated from Culicoides peregrinus vector from Tamil Nadu state. After appearance of 75% cytopathic effect in BHK-21cell culture, viral nucleic acid (dsRNA) was extracted. RNA-PAGE analysis showed the BTV specific characteristics migration pattern (3: 3: 3: 1) of viral nucleic acid. Furthermore, viral nucleic acid was subjected to cDNA synthesis for ns1 and vp2 gene based RT-PCR analysis. The ns1 gene RT-PCR, specific cytopathic effect and migration pattern in RNA-PAGE confirmed the sample as BTV. The vp2 gene serotype specific RT-PCR identified the isolate as BTV serotype 23. The study suggested that Culicoides peregrinus could be a potential vector which plays important role in BTV transmission in southern India.

Geminivirus: Indian Scenario

Geminiviruses are a diverse group of plant virus which infects several crop species in India. Based on host range, insect vector, and genome organization, geminiviruses are classified into four genera: Begomovirus, Curtovirus, Topocuvirus, and Mastrevirus. Begomovirus are isolated from many of the Indian crop plants such as tomato, chilies, cotton, pulses, papaya, cucurbits, okra, etc. Begomoviruses are transmitted by whiteflies. They possess a bipartite genome of approximately 2.6 kb in size. Begomovirus genome may also possess satellite DNA molecules along with its genome. The infections of begomoviruses lead to significant economic loss to Indian crops. Therefore research on begomoviruses in India has been mainly focused on the molecular diagnosis of virus along with phylogenetic analyses, DNA replication, transgenic resistance, etc. Various techniques used for geminiviruses are antibody-based detection methods such as enzyme-linked immunosorbent assay (ELISA), nucleic acid hybridizations, dot blot, Southern blot, and polymerase chain reaction (PCR) along with direct visualization of virus using electron microscopy.