#sindbis

[bpod-bpod]

Change the Channel

Blocking the budding of viruses – when they wrap themselves in the host cell membrane and move off to spread the infection – could be a good strategy to stop them in their tracks. Researchers interested in alphaviruses – a type that includes chikungunya and Sindbis viruses – examined the role of a protein called 6K in virus assembly and spreading. They found that it functions as a channel to let molecules pass through membranes, like has been seen in proteins of HIV and influenza. This ion channel activity was needed for the formation of vacuoles in the cell, which promote the transport of proteins to the cell lining where they are incorporated into new virus particles (these proteins in red spread throughout the membrane, left, compared to clumping in the absence of 6K, right). Interfering with 6K action could be a new way to treat infection, and nip budding in the bud.

Written by Anthony Lewis

Image from work by Zeinab Elmasri and colleagues

Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, USA

Image originally published with a Creative Commons Attribution 4.0 International (CC BY 4.0)

Published in PLOS Pathogens, October 2022

You can also follow BPoD on Instagram, Twitter and Facebook

#science #biomedicine #chikungunya #sindbis #influenza #hiv #ion channel #viruses #vacuoles #immunofluorescence

[leedsomics]

Long term co-circulation of multiple arboviruses in southeast Australia revealed by xeno-monitoring of mosquitoes and metatranscriptomics

Arbovirus surveillance of wild-caught mosquitoes is an affordable and sensitive means of monitoring virus transmission dynamics at various spatial-temporal scales, and emergence and re-emergence during epidemic and interepidemic periods. A variety of molecular diagnostics for arbovirus screening of mosquitoes (known as xeno-monitoring) are available, but most provide limited information about virus diversity. PCR-based screening coupled with metatranscriptomics is an increasingly affordable and sensitive pipeline for integrating complete viral genome sequencing into surveillance programs. This enables large-scale, high-throughput arbovirus screening from diverse samples. We collected mosquitoes in CO2-baited light traps from five urban parks in Brisbane from March 2021 to May 2022. Mosquito pools of [≤]200 specimens were screened for alphaviruses and orthoflaviviruses using virus genus-specific primers and reverse transcription quantitative PCR (qRT-PCR). A subset of virus-positive samples was then processed using a mosquito-specific ribosomal RNA depletion method and then sequenced on the Illumina NextSeq. Overall, 54,670 mosquitoes, representing 26 species were screened in 382 pools. Thirty detections of arboviruses were made in 28 pools. Twenty of these positive pools were further characterised using meta-transcriptomics generating 18 full-length genomes. These full-length sequences belonged to four medically relevant arboviruses: Barmah Forest, Ross River, Sindbis-like and Stratford viruses. Phylogenetic and evolutionary analyses revealed the evolutionary progression of arbovirus lineages over the last 100 years, highlighting long-distance dispersal across the Australian continent and continuous circulation characterised by constant turnover of virus lineages. http://dlvr.it/T4sKzP

[rebeccahaphotography]

Pernille Sindby photographed by me, January 2023

#business woman #flash photography #flash #photography advice #lifestyle photography #blonde #power woman #inspiring

[rnomics]

Proteomics-based determination of double stranded #RNA interactome reveals known and new factors involved in Sindbis virus infection [Article]

Viruses are obligate intracellular parasites, which depend on the host cellular machineries to replicate their genome and complete their infectious cycle. Long double stranded (ds)RNA is a common viral by-product originating during RNA virus replication and is universally sensed as a danger signal to trigger the antiviral response. As a result, viruses hide dsRNA intermediates into viral replication factories and have evolved strategies to hijack cellular proteins for their benefit. The characterization of the host factors associated with viral dsRNA and involved in viral replication remains a major challenge to develop new antiviral drugs against RNA viruses. Here, we performed anti-dsRNA immunoprecipitation followed by mass spectrometry analysis to fully characterize the dsRNA interactome in Sindbis virus (SINV) infected human cells. Among the identified proteins, we characterized SFPQ (Splicing factor, proline-glutamine rich) as a new dsRNA-associated proviral factor upon SINV infection. We showed that SFPQ depletion reduces SINV infection in human HCT116 and SK-N-BE(2) cells, suggesting that SFPQ enhances viral production. We demonstrated that the cytoplasmic fraction of SFPQ partially colocalizes with dsRNA upon SINV infection. In agreement, we proved by RNA-IP that SFPQ can bind dsRNA and viral RNA. Furthermore, we showed that overexpression of a wild type, but not an RNA binding mutant SFPQ, increased viral infection, suggesting that RNA binding is essential for its positive effect on the virus. Overall, this study provides the community with a compendium of dsRNA-associated factors during viral infection and identifies SFPQ as a new proviral dsRNA binding protein. http://rnajournal.cshlp.org/cgi/content/short/rna.079270.122v1?rss=1&utm_source=dlvr.it&utm_medium=tumblr

[houmannjust06]

Organic as well as Unnatural Brains: A shorter review of the particular interplay among AI along with neuroscience investigation.

Using structure-based design and style, we all produced a brand new semisynthetic compilation of spectinomycin analogs along with selective ribosomal inhibition and ideal narrow-spectrum antitubercular task. Inside several murine disease designs, these spectinamides had been properly tolerated, significantly lowered bronchi mycobacterial load and improved tactical. Within vitro scientific studies proven too little combination resistance using existing tb therapeutics, exercise towards multidrug-resistant (MDR) as well as extensively drug-resistant tb and a good medicinal report. Answer to their particular strong antitubercular properties had been his or her structurel modification for you to free yourself from your Rv1258c efflux water pump, that is upregulated throughout MDR ranges which is implicated inside macrophage-induced drug threshold. The particular antitubercular efficiency regarding spectinamides signifies that synthetic adjustments in order to Berzosertib classical prescription antibiotics can easily defeat the process involving intrinsic efflux pump-mediated weight along with expands opportunities for target-based tuberculosis medicine breakthrough discovery.To be able to overcome emerging coronaviruses, building safe and sound and effective programs to gauge virus-like protease routines along with the effectiveness of protease inhibitors can be a high concern. Right here, we manipulate a biosafety amount Two (BSL-2) chimeric Sindbis malware method to guage protease actions and the usefulness involving inhibitors aimed up against the papain-like protease (PLpro) of severe severe the respiratory system syndrome coronavirus (SARS-CoV), a biosafety stage Three or more (BSL-3) virus. Many of us designed Sindbis trojan to be able to coexpress PLpro plus a substrate, murine interferon-stimulated gene Fifteen (ISG15), and found that will PLpro mediates removal of ISG15 (deISGylation) from cellular healthy proteins. Mutation of the catalytic cysteine deposit of PLpro or inclusion of a new PLpro chemical clogged deISGylation inside virus-infected cellular material. Thus, deISGylation is often a sign regarding PLpro activity. An infection regarding alpha/beta interferon receptor ko (IFNAR(-/-)) rats using these chimeric infections said PLpro deISGylation action eliminated ISG15-mediated security during well-liked an infection. Notably, management of your PLpro inhibitor shielded these kind of these animals from deadly an infection, demonstrating the particular efficacy of an coronavirus protease chemical in the computer mouse product. However, this specific PLpro chemical wasn't sufficient to safeguard the particular these animals via deadly infection with SARS-CoV MA15, advising that more optimization from the supply and stability regarding PLpro inhibitors is necessary. We all prolonged your chimeric-virus podium to evaluate the actual papain-like protease/deISGylating action associated with Center Far east respiratory system symptoms coronavirus (MERS-CoV) to supply a small-animal model to judge PLpro inhibitors on this just lately come about virus. This specific podium has the potential to end up being universally adaptable to other well-liked and also cell phone nutrients which may have deISGylating routines. Relevance Evaluating viral protease inhibitors inside a small-animal style is often a crucial part of the trail in the direction of antiviral drug improvement. We all modified a new biosafety amount Only two chimeric trojan system to assist in look at inhibitors focused versus remarkably pathogenic coronaviruses. We all used this product to demonstrate the throughout vivo efficiency of an chemical of the papain-like protease involving serious acute respiratory system syndrome coronavirus. Furthermore, many of us show that the actual chimeric-virus system could be tailored to analyze the proteases associated with emerging individual bad bacteria, like Midst East breathing malady coronavirus. This system gives an essential tool to be able to rapidly assess the efficiency associated with protease inhibitors targeting active along with rising man infections, and also other nutrients capable of eliminating ISG15 via cellular protein.

#CDK2-IN-4 #PF-00299804 #LY2603618 #GSK2399872A #TEW-7197 #KPT-8602 #RG-7388 #NSC 122758 #NSC-100880 #LY2228820 #VPA #AG-120 #EDHS-206 #BC-2059 #BU-4061T #LY2606368 #Menin-MLL Inhibitor #GDC-0980 #ABBV-075 #NVP-AUY922 #Repertaxin #IPI-145 #FI-6934 #MK 733 #Berzosertib #Cycloheximide #Actinomycin D #Pelabresib #WC2031 #Nicotinamide Riboside

[health28]

Study warns of a potential Sindbis virus epidemic in 2022

Study warns of a potential Sindbis virus epidemic in 2022

In a recent Eurosurveillance study, researchers seek to raise awareness of a potentially impending Sindbis virus (SINV) epidemic due to its high incidence throughout Finland in 2021. Study: Sindbis virus outbreak and evidence for geographical expansion in Finland, 2021. Image Credit: Kateryna Kon / Shutterstock.com What is SINV? Although ribonucleic acid (RNA) viruses are not typically found in…

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[libertyforge]

There is so little understanding by those who say Ivermectin is an anti-parasitic drug, and it cannot, or it's ineffective or unknown in the treatment used for viruses, for Covid-19. From NCID, doctors, professors, pharmacists, mainstream news, keep hearing that. Well, IMO, they're very wrong. Ivermectin is not only an anti-parasitic drug. It's also a very effective anti-viral drug, and it works very well against single-stranded RNA viruses....including SARS-COV-2....and Dengue. On Viruses and Ivermectin Fact: The Zika virus is a single-stranded RNA virus. Fact: The dengue virus is a single-stranded RNA virus. Fact: Yellow river is a single-stranded RNA virus. Fact: Chikungunya is a single-stranded RNA virus. Fact: Sindbis is a single-stranded RNA virus. Fact: Semliki forest is a single-stranded RNA virus. Fact: Avian influenza A is a single-stranded RNA virus. Fact: Ivermectin is proven to be effective against Zika. Fact: Ivermectin is proven to be effective against Dengue. Fact: Ivermectin is proven to be effective against Yellow River. Fact: Ivermectin is proven to be effective against Sindbis. Fact: Ivermectin is proven to be effective against Semliki Forest. Fact: Ivermectin is proven to be effective against Avian influenza A. Fact: Human grade ivermectin is effective against all these as it can inhibit the replication process, which they all share. Fact: SARS-CoV-2 is a single-stranded RNA virus. Fact: SARS-CoV-2 shares the same replication process. https://ift.tt/3eVrLoR via Facebook https://ift.tt/3zcbjtH

#IFTTT #Facebook

[wapiti3]

The vertebrate zoology of Sind

By Murray, James A. Publication info London :Richardson,1884. Contributor: Smithsonian Libraries BIODIV LIBRARY

#scientific illustration #herpetology #lizards #snakes #reptiles #india #natural history #illustrations

[cheaponlinemedicinesblog]

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A partnership between Japan’s Kitasato Institute and Merck & Co. produced Ivermectin in the 1970s. Due to its ability to combat both internal and external parasites, Ivermectin 6mg tablets in USA quickly became a blockbuster drug in the veterinary field.

Ivermectin is used to treat humans for what?

This section describes some of the approved uses around the world.

The US market offers ivermectin in doses up to 200 mcg/kg once a year for the following indications:

a. Strongyloides stercoralis, an intestinal parasite that can cause severe systemic disease.

b. Onchocerciasis, also known as river blindness.

2. Ivermectin is also available in Europe as a single dose for Lymphatic filariasis and scabies.

3. Australia’s standard treatment for crusted scabies includes three doses of 200 mcg/kg administered within one month of diagnosis.

Another proven or potential use?

Ascaris lumbricoides and Trichuris trichura are two other intestinal parasites that Ivermectin may be effective against, although these parasites are not available for these purposes.

Off-label use is also popular for treating Tunga penetrans and head lice.

A promising area of research conducted by ISGlobal researchers involves applying Ivermectin 6mg tablets in USA at population levels to kill mosquitoes that feed on humans or animals and reduce malaria transmission.

How safe is ivermectin?

Ivermectin is a very safe drug when taken as prescribed for the current indications and dose. With an excellent safety profile, more than three billion treatments have been given to patients in the context of the Mectizan Donation Program alone to date. Most adverse reactions are mild, temporary, and caused by parasites dying rather than by the drug.

Invertebrates are the only animals with glutamate-gated chlorine channels.

Are higher doses of ivermectin safe?

In a study conducted by Guzzo et al., some volunteers safely received doses upto 2,000 mcg/kg (ten times the recommended dose for onchocerciasis).

According to Guzzo’s study, ivermectin does not cause transient visual disturbances, but other studies suggest that the drug may cause visual disturbances.

A dose of 600 mcg/kg (per day) for three days was administered safely by Smit et al.

Ivermectin has anti-viral properties

Definitely. There is evidence that Ivermectin 12mg tablets in USA  inhibits the replication of several RNA viruses, including:

Dengue

Zika

Yellow fever

West Nile

Chikungunya

Venezuelan equine encephalitis

Semliki Forest virus

Sindbis virus.

The virus is responsible for porcine reproduction and respiratory illnesses.

SARS-CoV-2

Why then do we need to study this?

There is no specific treatment available for COVID-19, which is a public health emergency of international concern. The excellent safety profile of Ivermectin and this fact warrant further investigation into its potential use. Similarly, applying the same conclusions directly to a living organism from a Petri dish is incorrect. Here are a few reasons why ivermectin may be more effective in vivo at lower doses than those described by Caly et al:

The immune system. Unlike a cellular culture, which is just a layer of cells in a Petri dish, a cellular culture lacks an immune system. Thus, it cannot defend itself against a virus alongside ivermectin.

A virus load. Ivermectin may understate the true efficacy of ivermectin because the virus-to-cell ratio may be too high in the petri dish, i.e. there may be an excess of virus-to-cells in the culture that would unbalance the fight in favour of the virus.

Immunomodulating effects. Ivermectin can modulate the immune system. Among the pathophysiologic characteristics of COVID-19 is an aggravated immune response.

A higher rate of lung penetration. Due to its lipophilicity, ivermectin accumulates in deep compartments, such as the lungs, although there is some disagreement regarding this.

Summary

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[didanawisgi]

Abstract

Ivermectin proposes many potentials effects to treat a range of diseases, with its antimicrobial, antiviral, and anti-cancer properties as a wonder drug. It is highly effective against many microorganisms including some viruses. In this comprehensive systematic review, antiviral effects of ivermectin are summarized including in vitro and in vivo studies over the past 50 years. Several studies reported antiviral effects of ivermectin on RNA viruses such as Zika, dengue, yellow fever, West Nile, Hendra, Newcastle, Venezuelan equine encephalitis, chikungunya, Semliki Forest, Sindbis, Avian influenza A, Porcine Reproductive and Respiratory Syndrome, Human immunodeficiency virus type 1, and severe acute respiratory syndrome coronavirus 2. Furthermore, there are some studies showing antiviral effects of ivermectin against DNA viruses such as Equine herpes type 1, BK polyomavirus, pseudorabies, porcine circovirus 2, and bovine herpesvirus 1. Ivermectin plays a role in several biological mechanisms, therefore it could serve as a potential candidate in the treatment of a wide range of viruses including COVID-19 as well as other types of positive-sense single-stranded RNA viruses. In vivo studies of animal models revealed a broad range of antiviral effects of ivermectin, however, clinical trials are necessary to appraise the potential efficacy of ivermectin in clinical setting.

#ivermectin #family medicine #covid-19 #sars-cov-2 #immunology #virology #fav #print this off later

[rebeccahaphotography]

Pernille Sindby photographed by me, January 2023

#business #flash photography #flash #photography #portrait #different portrait #real life photography

[suckhoevietnamsk]

Fucoidan hỗ trợ sức khỏe cho người bị viêm gan

Fucoidan thường có nhiều trong rong biển, có khả năng ức chế sự nhân lên của virus viêm gan C, có lợi cho bệnh nhân mắc viêm gan.

Gan là cơ quan nội tạng lớn nhất trong cơ thể, được ví như "nhà máy lọc máu chính". Gan còn đóng vai trò quan trọng trong quá trình chuyển hóa và một số chức năng khác như dự trữ, tổng hợp các chất, khử độc, miễn dịch.

Khi gan bị viêm làm ảnh hưởng đến khả năng hoạt động nhất là giải độc. Viêm gan B, C là những loại viêm gan thường gặp và tiến triển âm thầm. Theo báo cáo của Cục Y tế Dự phòng, ước tính nước ta có 10 triệu người nhiễm virus viêm gan B và gần một triệu người nhiễm virus viêm gan C. Cứ 100 người thì có đến 20 người nhiễm virus viêm gan B, 4 người nhiễm virus viêm gan C.

Một trong những thực phẩm có nhiều ứng dụng phòng ngừa và hỗ trợ sức khỏe cho người bị viêm gan như fucoidan. Fucoidan là một polysaccharide sulfat được chiết xuất từ rong nâu biển. Rong nâu có chứa fucoidan được tiêu thụ rộng rãi, có trong độ ăn uống ở Đông Á (nhất Trung Quốc, Nhật Bản và Hàn Quốc).

Nhiều nghiên cứu cho thấy fucoidan có một số hoạt tính sinh học bao gồm hỗ trợ kháng vi khuẩn, góp phần chống oxy hóa và nhất là khả năng chống viêm. Fucoidan có lợi trong các lĩnh vực viêm nhiễm, điều hòa miễn dịch và ức chế enzym.

Rong nâu thường có trong Rong nâu thường có trong chế độ ăn của người Nhật Bản. Ảnh: freepik.

Một nghiên cứu của Naoki Mori và đồng sự tại Đại học Ryukyus, Nhật Bản đươc đăng tải trên Trung tâm Thông tin Công nghệ sinh học Quốc gia Mỹ năm 2012 cho thấy, fucoidan ức chế sự nhân lên của virus viêm gan C (HCV) nhưng không có tác dụng gây độc tế bào. Nghiên cứu được thực hiện trên 15 người trong 12 tháng. Sau khi dùng fucoidan, những bệnh nhân nhiễm HCV có xu hướng làm giảm nồng độ alanin aminotransferase trong huyết thanh, tương quan với việc giảm đáng kể HCV. Nghiên cứu kết luận, fucoidan ức chế sự nhân lên của HCV nhưng không có tác dụng gây độc tế bào.

Naoki Mori cho biết, fucoidan còn có lợi cho những bệnh nhân bị nhiễm virus viêm gan B mãn tính. Fucoidan có thể ức chế hiệu quả sự nhân lên của HBV trong cơ thể sống và trong ống nghiệm. Điều trị bằng fucoidan cũng làm giảm lượng protein lõi HBV và sự bài tiết HBsAg và HBeAg. Tuy nhiên, cơ chế tác động của fucoidan đối với sự biểu hiện HBV cần được nghiên cứu thêm.

Cũng theo nghiên cứu này, các polysaccharid sulfat bao gồm cả fucoidan, cũng được báo cáo là có khả năng ức chế sự nhân lên của các loại virus như virus herpes simplex, virus sindbis, virus suy giảm miễn dịch ở người, virus parainfluenza loại II và virus sốt xuất huyết. Vì fucoidan không có độc tính hoặc không gây kích ứng ở người nên nó cũng có thể hữu ích như một chất chống HCV.

Bệnh viện Đại học Quốc gia Seoul - Hàn Quốc đã nghiên cứu ứng dụng của Fucoidan trong rong nâu. Kết quả cho thấy Fucoidan góp phần đẩy mạnh quá trình tự chết của tế bào ung thư gan, hỗ trợ làm chậm quá trình tự chết của tế bào gan khỏe mạnh. Tác động kép này phần nào góp phần giúp bảo tồn chức năng gan, ngăn khối u di căn nhanh hơn.

Fucoidan được ứng dụng trong lĩnh vực sức khỏe, có trong các thực phẩm chức năng với hoạt tính góp phần kháng virus. Một số sản phẩm có chứa fucoidan hỗ trợ cải thiện cho bệnh nhân mắc các bệnh về mãn tính về gan, ung thư.

[medicinevilla]

Why Ziverdo Kit is the Effective Tripple Therapy to Treat COVID-19?

Many experts’ medical experts and directors suggested that FDA and TGA approved Ivermectin i.e ziverdo kit which utilizes consistently in his clinic, has shown positive outcomes for COVID-19 and will be considered promptly to battle the pandemic.

 Ivermectin was found in the 1970s and is in the world Health Organization (WHO) rundown of fundamental meds. utilized an identical philosophy with the COVID-19 Ivermectin Triple Therapy as he utilized when he built up the world's first remedy for peptic ulcers saving numerous lives throughout the planet. This blend of three endorsed "off the rack" medications may be the answer for the COVID-19 emergency.

 Our older are at absolute best danger and this is frequently a truly protected alternative particularly once we don't have whatever else except for ventilators. Additionally, our forefront laborers merit more security with a deterrent medicine this way, and as a crisis treatment if that they test positive. Buy ziverdo kit online to get rid of COVID-19.

 An Ivermectin tablet can cost as little which could make it far and away from the most cost-effective, safest, and fastest cure for peoples and therefore our country's economy.

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 Research has driven him to a triple treatment of Ivermectin, zinc and an anti-toxin (Ziverdo Kit) which are all TGA and FDA affirmed which may be the quickest and most secure because of stop the Victorian episode inside 6 two months.

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 Ziverdo Kit is recommended for Covid patients as per recent studies and has shown significant results. It consists of Ivermectin 12mg, Zinc Acetate 50mg & Doxycycline 100mg a lethal Combination to fight against Covid-19.

 the key antiviral research project literature and identified the mixture of three drugs that are in chemists immediately and may be prescribed by doctors immediately. The tablets are often taken reception as a preventive treatment by high-risk individuals, or by those that test positive to minimize the need for hospitalization at the upper curative dose.

 A few investigations detailed antiviral impacts of ivermectin on RNA infections like Zika, dengue, yellow jack, West Nile, Hendra, Newcastle, Venezuelan equine encephalomyelitis, chikungunya, Semliki Forest, Sindbis, Avian flu A, Porcine Reproductive and Respiratory Syndrome, Human immunodeficiency infection type 1, and serious intense respiratory condition Covid 2.

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[cloeadventurelog]

5/3/18

PAMPHLET TIME Adenovirus Poliovirus HIV Sindbis VSV Flu SV40 Polyomavirius Herpes Simplex virus type 1 Poxvirus Bacteriophages

[lupine-publishers-aoics]

Lupine Publishers|

Follow on Linkedin : https://www.linkedin.com/company/lupinepublishers Follow on Twitter   :  https://twitter.com/lupine_online For more Lupine Publishers Open Access Journals Please visit our website: http://lupinepublishers.us/ For more Open Access Journal on Chemistry articles Please Click Here: https://lupinepublishers.com/chemistry-journal/ To Know More About Open Access Publishers Please Click on Lupine PublishersMolecular Characterization of Chikungunya Virus in Serum- Relevance for Disease Management

Lupine publishers| Polymer chemistry

Abstract

Background: Chikungunya Virus is a single stranded RNA virus of the genus alphavirus. The transmission of this virus occurs by the bite of infected mosquitoes. It is characterized by an abrupt onset of fever frequently accompanied by joint pain.

Aim: Molecular Characterization of Chikungunya Virus in Serum- Relevance for Disease Management.

Methods and Materials: 30 cases were considered having symptom of fever suspected to have chikungunya. RNA was isolated from magnetic beads methods which were further studied for Real Time-PCR (RT-PCR).

Results: Among the 30 cases suspected for chikungunya fever, only 18 were positive for the same. Hence, the prevalence was calculated to be 60%. About 39% of the positive cases were in the age range of 21-40 years, which is the active age group of the total positive cases, 66.66% were males and 33.33% were females. The virus can be even transmitted together as a co-infection. Patients infected with chikunguniya virus are more likely to experience symptoms of high fever, severe polyarthralgia and rash with lymphopenia.

Keywords: Chikunguniya; Real time polymerase chain reaction; Aedes aegypti; Aedes albopictus

Abbrevations: CHIK V: Chikungunya Virus; CMRL: Central Molecular Research Laboratory; SGRRIM & HS: Shri Guru Ram Rai Institute of Medical & Health Science; CDNA: Complementary DNA; MRNA: Messenger RNA

Introduction

Chikungunya Virus (CHIKV) is an alphavirus with a positive sense single-stranded RNA genome of approximately 11.6kb, 60- 70nm diameter capsid and a phospholipid envelope. It is sensitive to desiccation and to temperatures above 58°C. It is a member of the Semliki Forest Virus complex. The transmission of this virus occurs with the bite of infected mosquitoes of the genus Aedes. In addition, blood born transmission is possible [1-3]. The incubation period ranges from 3 to 12 days. The onset is usually abrupt and the acute stage is characterized by sudden high fever, incapacitating arthralgia, myalgias and skin rash. Chronic arthritis may develop in the patients and is associated with fever, asthenia and exacerbation of arthralgia, inflammatory polyarthritis, and stiffness [4,5]. The Alphavirus group comprises of 28 viruses, six of which can cause human joint disorders-namely chikungunya virus, o’nyong-nyong virus (central Africa), Ross River and Barmah Forest viruses (Australia and the Pacific), Sindbis virus (cosmopolitan), and Mayaro virus (South America, French Guyana). These alphaviruses share certain antigenic determinants [6-9].

Materials and Methods

30 clinical samples were considered in this study. Blood samples were taken from the suspected cases of Chikungunya from the different departments of Shri mahant Indiresh Hospital, dehradun (Uttarakhand) and further processed at Central Molecular Research Laboratory (CMRL), Shri Guru Ram Rai Institute of Medical & Health science (SGRRIM & HS), Patel nagar, dehradun (U.K.) for the molecular characterization of Chikungunya Virus.

Collection of EDTA Blood & Separation of Serum/Plasma

Whole blood samples were centrifuged at 5,000rpm for 10- 15 minutes at 4°C. The serum was separated from blood & then subjected for nucleic acid extraction. Then Isolation of RNA for the detection of chikungunya virus was done by magnetic beads method where Magnetic carriers bearing an immobilized affinity or hydrophobic ligand or ion exchange groups or magnetic biopolymer particles having affinity to the isolated structure are mixed with the sample containing target compounds. The specimens with target nucleic acid are processed by lysis buffer and then the nucleic acids are efficiently bound to the specifically modified magnetic beads. The beads were removed from the solution and attached to the tube wall manually using a magnetic separator stand. After washing, purification and elution, ultra-purified RNA can be obtained. After RNA was obtained master mix was prepared for chikungunya RNA for qualitative detection by Real Time PCR. Reverse transcription was done (RT-PCR) for the extracted RNA specimens. In this method, RNA was first transcribed into complementary DNA (cDNA) by reverse transcriptase from total RNA or messenger RNA (mRNA). The cDNA is then used as the template for the qPCR reaction.

Results

Among the 30 cases suspected for chikungunya fever, only 18 were positive for the same. Hence, the prevalence was calculated to be 60%. About 39% of the positive cases were in the age group ranging in between 21-40 years, which is the active age group of the total positive cases, 66.66% were males and 33.33% were females. Ct value in molecular profiling indicates that the sample is positive for chikungunya viral genome whereas no Ct value is indication of negative results for chikungunya. Ct value for internal control was in the range of 14-17 shows that the result is valid (Figure 1).

Figure 1: Amplification curves in Real time PCR for Chikungunya virus.

Discussion and Conclusion

Chikungunya (CHIKV) is a mosquito-borne disease caused by an RNA alphavirus of the Togaviridae family. The main mosquito vectors are the aggressive Aedesaegypti and Aedesalbopictus. The most common presentation of CHIKV is acute onset of fever and polyarthralgia, sometimes followed by a macupopular rash. Other associated symptoms can include headache, myalgia, nausea and vomiting. These symptoms typically occur 3 to 7 days after the mosquito bite and generally last 7 to 10 days. Rare complication of CHIKV can include uveitis, retinitis, myocarditis, hepatitis, nephritis, bullos skin lesions, hemorrhage, meningoencephalitis. The clinical manifestations of CHIKV can be very similar to dengue fever, which is transmitted by the same species of the mosquito. The 2 viruses can be even transmitted together as a co-infection. Patients infected with CHIK are more likely to experience symptoms of high fever, severe polyarthralgia and rash with lymphopenia, whereas patients with dengue fever are more likely to have symptoms of hemorrhage, shock and death with associated laboratory derangements of neutropenia and thrombocytopenia. However, because of the close similarities, dengue fever should be strongly considered in the differential diagnosis of patients suspected with CHIK. Conformational testing for CHIK can be performed in one of the three ways. These methods include viral culture if tested within first three days of illness, PCR in the first eight days, or antibody serology after the first three days of illness. Other laboratory abnormalities may include lymphopenia, thrombocytopenia, elevated creatinine and elevated hepatic transaminases. Treatment is primarily supportive. The preventive measure among the individual level included the use of mosquito repellents like coils, body creams, mosquito nets, etc. At the community level, it is important to ensure that there are no collections of water in household vessels or around dwelling places. The usual recommendation is that on every fifth day, all the vessels, which contain water, should be emptied; this observance of a dry day breaks the life cycle of the mosquito [10].

From above discussion, we conclude that, Chikungunya fever is self-limiting, the morbidity can be very high in major outbreaks, resulting in heavy social and economic tolls. The prevention of the disease requires a planned approach, besides knowledge and awareness on the early warning signs. An integrated vector management through the elimination of the breeding sites, the use of anti-adult and anti-larval measures and personal protection will contribute to the prevention of outbreaks. A community empowerment and mobilization is crucial for the prevention and control of Chikungunya.

Acknowledgement

The authors are grateful to Honorable Chairman, Shri Guru Ram Rai Education Mission for his kind support, guidance and favor.

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I like the slices being shown with the corresponding cutaway of the reconstructed particle