Wednesday, March 25, 2020
Analysis of 'Feathers,' a Short Story by Raymond Carver American poet and author Raymond Carver (1938 - 1988) is one of those rare writers who is known, likeÃ Alice Munro, primarily for his work in the short story form. Due to his economical use of language, Carver is often associated with a literary movement known as minimalism, but he himself objected to the term. In a 1983 interview, he said, Theres something about minimalist that smacks of smallness of vision and execution that I dont like. Feathers is the opening story of Carvers 1983 collection, Cathedral, in which he began to move away from the minimalist style. Plot of Feathers SPOILER ALERT: If you dont want to know what happens in the story, dont read this section. The narrator, Jack, and his wife, Fran, are invited to dinner at the home of Bud and Olla. Bud and Jack are friends from work, but no one else in the story has met before. Fran is not enthusiastic about going.Ã Bud and Olla live in the country and have a baby and a pet peacock. Jack, Fran, and Bud watch television while Olla prepares dinner and occasionally tends to the baby, who is fussing in another room. Fran notices a plaster cast of very crooked teeth sitting on top of the television. When Olla enters the room, she explains that Bud paid for her to have braces, so she keeps the cast to remind me how much I owe Bud. During dinner, the baby begins fussing again, so Olla brings him to the table. He is shockinglyÃ ugly, butÃ Fran holds himÃ and delights in him in spite of his appearance. The peacock is permitted inside the house and plays gently with the baby. Later that night, Jack and Fran conceive a child even though they had not previously wanted children. As the years pass, their marriage sours and their child demonstrates a conniving streak. Fran blames their problems on Bud and Olla even though she saw them only on that one night. Wishes Wishes play a prominent role in the story. Jack explains that he and Fran regularly wished out loud for things we didnt have, like a new car or the chance to spend a couple of weeks in Canada. They dont wish for children because they dont want children. It is clear that the wishes arent serious. Jack acknowledges as much when he describes approaching Bud and Ollas house: I said, I wish we had us a place out here. It was just an idle thought, another wish that wouldnt amount to anything. In contrast, Olla is a character who has actually made her wishes come true. Or rather, she and Bud together have made her wishes come true. She tells Jack and Fran: I always dreamed of having me a peacock. Since I was a girl and found a picture of one in a magazine. The peacock is loud and exotic. Neither Jack nor Fran has ever seen one before, and it is much more dramatic than any of the idle wishes theyve been making. Yet Olla, an unassuming woman with an ugly baby and teeth that needed straightening, has made it a part of her life. Blame Though Jack would place the date later, Fran believes their marriage began to deteriorate precisely on the night they had dinner at Bud and Ollas, and she blames Bud and Olla for it. Jack explains: Goddamn those people and their ugly baby, Fran will say, for no apparent reason, while were watching TV late at night. Carver never makes it clear exactly what Fran blames them for, nor does he make it clear exactly why the dinner gathering inspires Jack and Fran to have a baby. Perhaps its because Bud and Olla seem so happy with their strange, squawking-peacock, ugly-baby lives. Fran and Jack dont think they want the particulars - a child, a house in the country, and certainly not a peacock - yet perhaps they find they do want the contentedness that Bud and Olla seem to have. And in some ways, Olla does give the impression that her happiness is a direct result of the particulars of her situation. Olla compliments Fran on her naturally straight teeth while she herself had required braces - and Buds devotion - to fix her crooked smile. At one point, Olla says, You wait until you get our own baby, Fran. Youll see. And as Fran and Jack are leaving, Olla even hands Fran some peacock feathers to take home. Gratitude ButÃ Fran seems to be missing one fundamental element that Olla has: gratitude. When Olla explains how grateful she is to Bud for straightening her teeth (and, more generally, giving her a better life), Fran doesnt hear her because she is picking through the can of nuts, helping herself to the cashews. The impression is that Fran is self-centered, so focused on her own needs that she cant even hear someone elses expression of gratitude. Similarly, it seems symbolic that when Bud says grace, Olla is the only one who says amen. Where Happiness Comes From Jack does note one wish that came true: What I wished for was that Id never forget or otherwise let go of that evening. Thats one wish of mine that came true. And it was bad luck for me that it did. The evening seemed very special to him, and it left him feeling good about almost everything in my life. But he and Fran may have miscalculated where that good feeling was coming from, thinking it came from having things, like a baby, rather than feeling things, like love and appreciation.
Friday, March 6, 2020
Convert Frequency to Wavelength Worked Example Problem This example problem demonstrates how to find the wavelength of light from the frequency. Frequency vs Wavelength The wavelength of light (or other waves) is the distance between subsequent crests, valleys, or other fixed points. The frequency is the number of waves that pass a given point in one second. Frequency and wavelength are related terms used to describe electromagnetic radiation or light. One simple equation is used to convert between them: frequency x wavelength speed of light Ã » v c, when Ã » is wavelength, v is frequency, and c is the speed of light so wavelength speed of light / frequency frequency speed of light / wavelength The higher the frequency, the shorter the wavelength. The usual unit for frequency is Hertz or Hz, which is 1 oscillation per second. Wavelength is reported in units of distance, which often ranges from nanometers to meters. Conversions between frequency and wavelength most often involve wavelength in meters because thats how most people remember the speed of light in a vacuum. Key Takeaways: Frequency to Wavelength Conversion Frequency is how many waves pass a defined point per second. Wavelength is the distance between successive peaks or valleys of a wave.Frequency multiplied by wavelength equals the speed of light. So, if you know either the frequency or the wavelength you can calculate the other value. Frequency To Wavelength Conversion Problem The Aurora Borealis is a night display in the Northern latitudes caused by ionizing radiation interacting with the Earths magnetic field and the upper atmosphere. The distinctive green color is caused by the interaction of the radiation with oxygen and has a frequency of 5.38 x 1014 Hz. What is the wavelength of this light?Solution:The speed of light, c, is equal to the product of the wavelength, lamda;, and the frequency, Ã ½.ThereforeÃ » c/Ã ½Ã » 3 x 108 m/sec/(5.38 x 1014 Hz)Ã » 5.576 x 10-7 m1 nm 10-9 mÃ » 557.6 nmAnswer:The wavelength of the green light is 5.576 x 10-7 m or 557.6 nm.
Wednesday, February 19, 2020
Asset Classification Policies and Procedures - Essay Example Asset Classification Policies and Procedures For information to be handled properly, employees are required to have a working knowledge of the categorization of information into the three provided categories. Employees should be able to categorize the information before forwarding it further. If at some point, employees are confused about properly categorizing the information, the proper course of action is to classify it as confidential while an appropriate supervisor would later review and properly classify. Public Data- As the name suggests, such data is usually open to the public and is easily available. Disclosure of such data does not put the firm in any kind of risk; however certain controls are required to be enforced on such data to prevent modification or destruction of the data by unauthorized parties; Sensitive Data- Data is classified as sensitive data when disclosures of such information publically can result in potential risk for the organization or its people. Such information may be provided to others on a discretionary basis and under the supervision of the data owner. Confidential Data Ã¢â¬â Confidential data is the most sensitive data within the organization and unauthorized disclosure of such information can result in significant risk for the firm. The highest level of security and control are applied on such information. The System Impact level will determine the impact of activities on the system on a scale of one to five with five being the most crucial impact and one having the least crucial impact.
Tuesday, February 4, 2020
TOURIST ATTRACTION SITE VISIT - Essay Example These hotels provide amazing facilities to tourists. I asked organizing committee about procedures for purchasing tickets for soccer season. They replied, online tickets for complete season can be purchased at cheap rates and it is also possible to select the place of own choice for watching the matches in stadium. I asked organizers that what all facilities are available at the stadium for watching soccer competitions. They replied, they have introduced great facilities like, bathrooms, food, cheerleaders, sound effects and screens for adding all sorts of fun in the events. Who all are tough teams in the event? Mayor said, Ã¢â¬Å"Houston DynamoÃ¢â¬ and Ã¢â¬Å"DallasÃ¢â¬ are the best teams of the event. I asked how many stadiums are available for hosting the event. Mayor counted four stadiums including a newly constructed state of the art stadium. What is tailgating in Houston? Mayor said that you have snacks and hotdogs before start of the match is tailgating. What are the other tourist destinations in Houston? Mayor replied, space Centre and downtown aquarium are worth visiting places. Organizing committee told me to visit Orange Avenue as well. This avenue provides free souvenirs, food, shirts and a band plays there to entertain people. I am enjoying the opening games 2014 more than my expectations. It is so fascinating and colourful for people of any place in the world. I heard about the Houston and soccer events from my friend and then visited online sources. After coming here, I am not ready to leave this place now. Event is so organized that tourists do not feel any problem in their activities. There is one problem that spectators cannot buy single ticket for a match and they have to buy for complete season. Organizers need to look into this matter. Moreover, stadiums with more capacity should be constructed for accommodating huge crowds. Dynamo started the season against Ã¢â¬Å"New England RevolutionÃ¢â¬ on 8 Mar 2014. There is a long
Monday, January 27, 2020
Cahi-DRB and DQB1 Alleles in Sirohi Goat Genetic diversity of DRB and DQB genes of caprine MHC class II in Sirohi goat G. R. Gowane, Najif Akram,Ãâ S.S. Misra, Ved Prakash and Arun Kumar Running Head: CahiDRB and DQB1 alleles in Sirohi goat ABSTRACT Objective of the study was to assess the genetic diversity of the Sirohi goat for DRB and DQB1 loci and to study their association with antibody response induced by the Peste des petits ruminants (PPR) vaccine. A total of 360 Sirohi kids were studied using Single Stranded Confirmation Polymorphism (SSCP) followed by Sequence Based Typing (SBT)-PCR for DRB and DQB1 diversity. C-ELISA was used to assess immune response post PPR vaccination. Study revealed rich diversity of MHC region. A total of 18 DRB and 15 DQB1 alleles were obtained. Allele DRB*0104 and allele DQB1*0101 were most common. All the alleles reported are new. Study revealed variability in DRB and DQB1 region not only at nucleotide but also at amino acid level with high Wu-Kabat index. A total of 16 out of 89 amino acid residue sites had more than 3 amino acid substitutions in DRB. Similarly, 19 out of 86 residue sites in DQB1 had more than 3 amino acid substitutions. Positive evolutionary selection was evident in Sirohi for MHC region. Non-significant association of DRB and DQB1 genotypes with PPRV vaccine response revealed complexity of the phenotype and importance of other factors for vaccine response. Rich diversity of DRB and DQB1 gene reflects the fitness of the population and importance of this locus for future selection programs. Keywords: Cahi-DRB, Cahi-DQB1, Major histocompatibility complex, Vaccine response 1. Introduction Major Histocompatibility Complex (MHC) of goats is polymorphic. A few of the genes such as Caprine Leukocyte Antigen (Cahi)-DRB and Cahi-DQB1 from this complex are recently being investigated for their polymorphism and further potential association with important diseases of goat. The class II antigens encoded by MHC class II genes bind to processed peptides from extracellular antigens and present them to epitope specific CD4+ T lymphocytes. Cahi-DRB exon 2 is polymorphic so is Cahi-DQB1, due to their importance in antigen binding groove formation and evolutionary importance in antigen capture and presentation. Peptide binding site (PBS) in goat is partly coded by DRB and DQB gene. This PBS has several pockets which are highly variable and accommodate the side chains of the bound peptide. A non-synonymous change in the nucleotide sequence of the MHC DRB or DQB1 gene can substantially substitute the coding amino acid and ultimately bring conformational change in the binding groove so as to affect the efficiency of the protein to present the antigen efficiently for further processing. Several reports exists which link the variability in DRB alleles in cattle, sheep and other mammals to resistance or susceptibility to diseases. Herrmann-Hoesing et al.  reported that Ovar-DRB1 alleles contribute as a host genetic factor that control provirus level in sheep. Significant association of DRB1 alleles with susceptibility and resistance to Ovine pulmonary adenocarcinoma (OPA) was reported by Larruskain et al.  in sheep. However as far as studies on goat are concerned, there are very few caprine DRB and DQB1 sequences in Gene Bank. Similarly, there is a scarcity of research database for allelic association of DRB and DQB alleles with disease resistance or susceptibility in goat. It is for no surprise that even the IPD-MHC database has no space dedicated for goat MHC. Amills et al.  assessed the genetic variability in DRB of goat. This was followed by a few report s [4-8] to characterize DRB locus (exon 2 of DRB) in goat. Amills et al. (2004) also characterized DQB1 locus in goat, however not much work  has been carried out since then for its genetic polymorphism. Genetic variability in response to vaccination is likely to become an even more significant factor in designing ideal vaccines . The genes identified might also be important for disease resistance traits, and could potentially provide the tools to select good responders opening the doors for potential implications in future selection programme [11,12]. The Peste des petits ruminant (PPR) being the plague of small ruminants pose heavy threat to the rural economy of India. It is caused by a PPR virus (PPRV) of the genus Morbillivirus within the family Paramyxoviridae. India constitutes a great diversity of small ruminants with 135.17 million goat and 65.07 million sheep (19th Livestock census) . In PPRV endemic regions including India, control measures involve regular vaccination with live attenuated PPR virus vaccine of lineage IV, which has high antigenic stability and induce long term immune response . Currently, three live attenuated PPR vaccines (Sungri/96, Arasur/87 and CBE/97 stains) are available in India for prevention of this disease, of which, Sungri/96, developed by ICAR-Indian Veterinary Research Institute (IVRI), Mukteswar has undergone extensive field trial [15-17]. It is possible that the vaccine induced protection across individuals is not homogenous, wherein, vaccine gives a complete protection for a proportion of individuals while rest acquire only incomplete (leaky) protection of varying magnitude . Variable vaccine response in the population has been reported for several diseases in humans as well as animals [19-27]. Role of host genetics and other non-genetic factors in variation to vaccine response especially for PPR vaccine has not been studied till today in details. The importance of host genetics in vaccine response studies is important as genetic variability may influence vaccine response and hence confound vaccine efficacy studies. Objective of the present study is to decipher the Cahi-DRB and Cahi-DQB1 polymorphisms in detail using sequence based typing polymerase chain reaction (SBT-PCR) and to associate the variation obtained with PPR vaccine elicited immune response in Sirohi goat kids maintained at the farm condition in semi-arid region of India. 2. Materials and Methods 2.1 Animals The study population was a flock of purebred Sirohi goats. The flock was located at ICAR-Central Sheep Wool Research Institute, Avikanagar in the semi-arid region of Rajasthan, India at 75025Ã¢â ¬Ã ²E, 26018Ã¢â ¬Ã ²N, at an altitude of 320 m above mean sea level. The data for the experiment involved 360 Sirohi goat kids. All the animals under the study belonged to same age group, i.e. weaner with mean age at vaccination 142.43 days (SD = 14.67). All the animals in this flock were kept under semi-intensive management system.Ã Concentrate mixture was offered ad libitum to suckling kids from 15 days of age till weaning (90 days). After 3 weeks of age till weaning, kids were sent for grazing for 3 h each in morning and evening, but not along with their dams. During the post-weaning period in addition to 8-10 h grazing and dry fodder supplementation, 300 g of concentrate mixture was provided in the evening hours after browsing. The grazing area consisted of forestland with natur al fodder trees like Khejri (Prosopis cineraria), Ardu (Ailanthus spp.), and Neem (Azadirecta indica). Bushes and surface vegetation including the improved pastures of Cenchrus ciliarisis are also available. Due to scarce grazing resources from March to June, the goats were supplemented with hay of Cenchrus, Cowpea, and Dolichos; pala leaves (Zizyphus) and fodder tree lopping. 2.2 Amplification and typing of DRB alleles Whole blood (1 ml) was collected aseptically from the jugular vein of lambs for DNA isolation (GenElute Blood Genomic DNA Kit, SIGMA) according to the manufacturers instructions. Exon 2 of the DRB gene was amplified from genomic DNA using the primers as suggested by Amills et al. , where DRB.1: 5-TATCCCGTCTCTGCAGCACATTTC-3 and DRB.2: 5-TCGCCGCTGCACACTGAAACTCTC-3 primers were used for amplifying 285 bp product. The reaction mixture of 50ÃÅ½Ã ¼l comprised of: 10X Taq Buffer (05ÃÅ½Ã ¼l), 25mM MgCl2 (03ÃÅ½Ã ¼l), 10mM dNTP (1ÃÅ½Ã ¼l), 20 pmol (1ÃÅ½Ã ¼l) of each primer, Taq DNA Polymerase (1IU), Template (1ÃÅ½Ã ¼l) and Nuclease Free Water (NFW) to make 50ÃÅ½Ã ¼l. The thermal profile was optimized for amplification of the DRB exon2 as follows: Initial denaturation (94Ã °C for 4 min), followed by 35 cycles (denaturation for 94Ã °C for 60 s, annealing at 66Ã °C for 60s and extension at 72Ã °C for 60s) and a final extension at 72Ã °C for 5 min. A single clear band of 285 bp on agarose gel (2%) was obtained. The amplified products were subjected to Single Stranded Confirmation Polymorphism (SSCP) for determination of the genotypic variation . The samples were then grouped according to various genotypes as obtained on the SSCP gel. The representative samples were then again amplified using the PCR protocol as above and purified PCR products (GenEluteÃ¢â¬Å¾Ã ¢ Gel Extraction Kit, SIGMA) were sequenced by BigDye (Applied Biosystems, USA) sequencing reaction that exploits di-deoxy chain termination principle. The PCR-Sequence Based Typing (PCR-SBT) was used for further analysis. The homozygous sequences obtained were assigned an allelic name using nomenclature system as suggested by Ballingall and Tassi  to suit IPD MHC nomenclature system. The heterozygote samples were re-sequenced after cloning (InsTAclone PCR Cloning Kit, Thermo Fisher) to obtain one allele that was subsequently used to deduce another allele in heterozygous sample. Novel alleles were cloned, sequenced and confir med at least thrice. The amino acids at pocket positions were determined from the nucleotide sequences of the alleles using EditSeq software package V5.0 . Alleles which were derived and not confirmed in SBT-PCR were not named. 2.3 Amplification and typing of DQB1 alleles Exon 2 of the DQB1 gene was amplified from genomic DNA using the primers as described by Amills et al. , where DQB-F: 5- CCC CGC AGA GGA TTT CGT G -3 and DQB-R: 5- ACC TCG CCG CTG CCA GGT -3 primers were used for amplifying 280 bp product having 8bp intron1, 270bp exon2 and 2bp intron2. The reaction mixture of 50ÃÅ½Ã ¼l comprised of: 10X Taq Buffer (05ÃÅ½Ã ¼l), 25mM MgCl2 (03ÃÅ½Ã ¼l), 10mM dNTP (1ÃÅ½Ã ¼l), 20 pmol (1ÃÅ½Ã ¼l) of each primer, Taq DNA Polymerase (1IU), Template (1ÃÅ½Ã ¼l) and Nuclease Free Water (NFW) to make 50ÃÅ½Ã ¼l. The thermal profile was optimized for amplification of the DQB exon2 as follows: Initial denaturation (94Ã °C for 4 min), followed by 35 cycles (denaturation for 94Ã °C for 45 s, annealing at 67Ã °C for 45s and extension at 72Ã °C for 45s) and a final extension at 72Ã °C for 5 min. A single clear band of 280 bp on agarose gel (2%) was obtained. The amplified products were subjected to Single Stranded Confirmation Polymorphism (SSCP ) for determination of the genotypic variation . The samples were then grouped according to various genotypes as obtained on the SSCP gel. The PCR-SBT approach was used for analysis. Alleles were named as per requirements of the IPD-MHC database , derived alleles were not named. 2.4 PPR Vaccination, Sampling and ELISA for detection of antibody against PPRV vaccine As part of the scheduled vaccination program, the animals were vaccinated (1 ml subcutaneous) with freeze dried live attenuated PPR virus (Sungri 96 strain) vaccine with PPR virus titre Ã¢â¬ °Ã ¥ 102.5 TCID50 (Raksha-PPR, Indian Immunologicals, India).Ã Whole blood was collected aseptically by jugular vein puncture from the kids at 28 days post vaccination (28DPV) for serum separation. Serum was collected and stored at Ãâ Ã¢â¬â¢20-Ã ¦C until testing. The ELISA for further analysis was done as described earlier . 2.6 Statistical Analysis The allelic frequencies, genotypic frequencies, phylogenetic analysis and residue substitution was studied using Microsoft excel package of the MS office (2010) and EditSeq (DNA STAR) software. Phylogenetic analysis was performed using MEGA 4.0, neighbor joining method. To assess the effect of genotype on vaccine response (observed PI values), a General Linear Model (GLM) was used that included Cohort (2 levels), Sex (2 levels), age at vaccination (continuous) as fixed effects along with either DRB or DQB1 genotype. All the above analyses were performed using a statistical package SPSS . 2.7 The dn/ds ratio and Wu Kabat variability index The frequencies of non-synonymous (dn) versus synonymous (ds) substitutions were calculated by the method of Yang and Nielsen  with the help of software PAML 4 . The Wu Kabat variability index with respect to amino acids at peptide binding pockets was calculated using the formula given by Wu and Kabat . Index =Ã Ã Ã The number of different amino acids occurring at a given positionÃ Ã Ã Ã Ã Ã Ã Ã Ã Ã Ã Ã Ã Ã Frequency of the most common amino acid at the position Where, frequency of the common amino acid is obtained as number of times the most common amino acid occurs divided by the total number of protein examined. 3. Results and Discussion 3.1 Genetic variability for DRB Sirohi goat kids (N=360) were typed for DRB exon 2. A total of 18 new alleles were obtained after analysis in the population using SBT-PCR approach (Table 1). Out of the 18 alleles, 12 alleles were confirmed by cloning and sequencing, however 6 were derived using SBT-PCR. All 12 alleles were new and named as per the requirements of the Immuno-Polymorphism Database (IPD) following guidelines . Alleles were Cahi-DRB*0701Ã Ã (accession no. KX431913), Cahi-DRB*0104Ã Ã (accession no. KX431914), Cahi-DRB*0402 (accession no. KX431915), Cahi-DRB*0102Ã Ã (accession no. KX431916), Cahi-DRB*0202Ã Ã (accession no. KX431917), Cahi-DRB*0501 (accession no. KX431918), Cahi-DRB*0401 (accession no. KX431919), Cahi-DRB*0103 (accession no. KX431920), Cahi-DRB*0203 (accession no. KX431921), Cahi-DRB*0101 (accession no. KX431922), Cahi-DRB*0201 (accession no. KX431923) and Cahi-DRB*0601 (accession no. KX431924). A total of 6 new alleles were derived using PCR-SBT approach, however not given names as per IPD-MHC nomenclature (N7, N11, N13, N16, N17, N18). Allele CahiDRB*0104 had highest frequency 29.72% followed by *0701 allele (22.64%), *0202 (13.89%) and *0102 (11.25%). In congruence with our finding, rich diversity of this region has been reported earlier in different goat breeds worldwide [3-8]. However, most of the studies were carried out using Restriction Fragment Length Polymorphism (RFLP) PCR, whereas, the current method of SSCP followed by SBT-PCR has more power to detect the genetic variability at DRB in goat. The ratio of non-synonymous (dN) to synonymous (dS) substitution for DRB gene in the Sirohi goat population was 3.24. This ratio was significantly greater than 1 indicating positive evolutionary selection for DRB gene in the present populations. However, the results are read with caution as the evidence is presumed and not absolute, due to lack of evidence for Capra species. It may be impossible to infer the selection pressure from the dN/dS measurement . In another study, 11.1 ratio for dN/dS was recorded in Peptide Binding Region of 12 Chinese indigenous goats for DRB*02 sequences . PBR being polymorphic, its importance is seen here. According to Simmons et al. , the long-term evolution, ancient and silent mutations also carried with translated mutations and became maintained in these regions. Pathogen-host interaction is complex, according to the Red Queen hypothesis , to be a part of this competition, diversity of MHC is important from hosts perspective. Plotting the phylogenetic tree for allelic relationship at nucleotide level revealed that the diversity was large (Fig 1). Clustering of the alleles revealed that some alleles tended to form closer clusters than others. Fig 2 reveals the amino acid variation between the alleles and it is seen that the population is polymorphic at coding region too, thus providing enough raw material for Sirohi goat population to tackle the pathogen variability. Study found that alleles DRB*0101, *0102, *0103 and *0104 had less than or equal to 4 codon change and hence clubbed together in one family. Derived allele *N18 also formed member of this group due to similarity of amino acid sequence. Similarly, alleles *0201, *0202, and *0203 had less than 4 amino acid differences. Alleles *0401 and *0402 had less than 4 amino acid differences, whereas, alleles *0501, *0601 and *0701 differed by more than 4 amino acid differences from each group. Predicted allele *N7 was related to *0701 due to similarity at amino acid level. Derived alleles *N11, *N13 and *N17 formed a group separate from others, similarly derived allele *N16 formed a different group. Phylogenetic analysis revealed that clustering based on nucleotide similarity and differences remained almost similar to clustering based on amino acid differences. 3.2 Genetic variability for DQB1 Sirohi goat kids (N=339) were typed for DQB1 exon 2. A total of 15 new alleles were obtained after analysis in the population using SBT-PCR approach (Table 1). Out of the 15 alleles, 13 alleles were confirmed by cloning and sequencing, however 2 were derived using SBT-PCR. All 13 alleles were new and named as per the requirements of the IPD . Alleles were CahiDQB1*0101 (Accession number KX431925), CahiDQB1*0201 (Accession number KX431926), CahiDQB1*0301 (Accession number KX431927), CahiDQB1*0302 (Accession number KX431928), CahiDQB1*0103 (Accession number KX431929), CahiDQB1*0501 (Accession number KX431930), CahiDQB1*0104 (Accession number KX431931), CahiDQB1*0701 (Accession number KX431932), CahiDQB1*0801 (Accession number KX431933), CahiDQB1*0102 (Accession number KX431934), CahiDQB1*070101 (Accession number KX431935), CahiDQB1*0502 (Accession number KX431936) andCahiDQB1*0202 (Accession number KX431937). A total of 2 new alleles were derived using PCR-SBT approach, however not given names as per IPD-MHC nomenclature (*N2, *N3). Allele CahiDQB1*0101 had highest frequency 27.22% followed by *070101 allele (13.02%), *N2 (11.69%) and *0201 (11.54%). Very high genetic diversity for this region has also been reported earlier [3, 31]. Similar diversity is also observed in sheep and cattle DQB1 region, however for goat there are very few studies. This study is the first report for DQB1 diversity in any Indian goat breed. To study the evolutionary stability or instability of the DQB1 region in Sirohi goat, the ratio of non-synonymous (dN) to synonymous (dS) substitution for Sirohi goat has been studied. We found that the ratio was 1.08. Yakubu et al.  reported a ratio of 2.14 in Nigerian goat breeds.Ã Results reveal balancing selection in favour of variability at DQB1 in Sirohi goat. Phylogenetic analysis for alleles reported that the diversity at nucleotide level was large (Fig 1). There was a clustering of alleles for their nucleotide substitutions and thus clubbing in one or the other family. Fig 3 reveals the amino acid variation between the alleles and it is seen that the population is polymorphic at coding region. Alleles DQB1*0101, *0102, *0103 and *0104 were in one group as they had less than 4 amino acid changes. Similarly, alleles *0201, *0202, and *0203 had less than 4 amino acid differences. Alleles *0201 and *0202 formed another family, alleles *0301 and *0302 formed separate family, and alleles 0501 and 0502 were clubbed together. It was seen that derived alleles N3 had similarity at amino acid level with allele *0201, indicative of synonymous substitution at nucleotide level.Ã Alleles *0701 and 070101 were in one family and they did not have a single amino acid substitution. However, they had synonymous differences at nucleotide level that resul ted in the no change at peptide level. Derived allele *N2 was related with *N3, however placed in separate group due to differences at amino acid level. 3.3 Association of DRB and DQB1 genes with PPRV vaccine elicited immune response Results of C-ELISA on sera samples at 28DPV revealed mean PI value of 69.99Ã ±0.42 (Min 13.32, max 91.60) with minimum PI 35.12 and maximum PI 98.82.Ã Ã AverageÃ ageÃ at vaccinationÃ wasÃ 142.43Ã Ã ±Ã 14.67Ã daysÃ withÃ minimumÃ ageÃ 93 daysÃ and maximumÃ ageÃ 164Ã days.Ã Variability in the vaccine response was evident in the lambs.Ã Frequency distribution of Ovar-DRB and DQB1 alleles revealed rich diversity amongst Sirohi goat. A total of 16 DRB genotypes and 16 DQB1 genotypes were observed to be present in the population of Sirohi goat flock. For association analysis, genotypes with >5 occurrences in the population (11 genotypes in DRB and 12 genotypes in DQB1) were only used to avoid biased estimates. Genotypic association analysis was carried out to assess the effect of genotype (Table 2) along with other environmental factors on vaccine response in Sirohi goat sheep. In the DRB group (N=299), Genotype I(DRB*0104-*0104) had highest frequency (30.10%) followed by genotype A(DRB*0701-*0701) 22.07% and genotype M(DRB*0202-*0202) 13.38%. In the DQB1 group (N=298), highest frequency was obtained for genotype E(DQB1*0801-*0801) 20.13%, followed by genotype J(DQB1*0301-*0101) 14.43% and genotype G(DQB1*0502-*0502) 11.41%. In the model that studied the effect of DRB genotype along with other environmental factors such as cohort, sex of the animal and age group, on vaccine response, explained 63.6% variation (R2=0.636). The genotypic association study revealed non-significant (P = 0.606) effect of genotype on 28DPV PI value, whereas significant effect of cohort and age at vaccination. However, ranking of genotypes revealed that the genotype L(DRB*0102-*0102) gave highest response for PPRV vaccination at 28th day (Table 2) followed by genotype J(DRB*0402-*0402) and A(DRB*0701-*0701).Ã Lowest response was obtained for the genotype E(DRB*0201-*0201) preceded by D(DRB*0101-*N13) and I(DRB*0104-*0104). It was noteworthy that alleles in high ranking genotypes were exclusive to low ranking genotypes. Effect of genotype was non-significant on the vaccine response, however, the trend was visible with increasing rank and declining mean PI for 28DPV (Table 2). The variability within DRB region of Sirohi goat population was calculated using the Wu-Kabat Variability Index (Table 3). The ability of a pocket to anchor a peptide is due to the electrostatic charges of the pocket region and electrostatic charges of the peptide . Out of several amino acid positions in DRB, a total of 16 different amino acid positions were polymorphic with three or more than 3 amino acid differences (residue 6, 21, 32, 35, 37, 52, 61, 62, 65, 66, 68, 69, 72, 73, 76 and 81). The region revealed Wu-Kabat index varying from 2.20 to 6.95. Highest index was observed at residue 6 (6.95%), followed by ÃÅ½Ã ²65 (6.41%) and ÃÅ½Ã ²73 (5.94%).Ã Present results corroborates with the earlier observations in sheep breeds [41, 42], where positive selection at important residues in DRB1 amino acid sequence was observed. In DQB1 group, again the inclusive model could explain 62% of the total variation in the 28DPV vaccine response trait (R2=0.62). The model included sex, age and cohort of the animal along with the DQB1 genotype. The effect of genotype was non-significant (P = 0.868), however, the effect of cohort and age at vaccination were highly significant (PDQB1*0104-*0701) gave highest response for PPRV vaccination at 28th day (Table 2) followed by Genotype E(DQB1*0801-*0801) and I(DQB1*0201-*0201). Lowest response was obtained for the genotype D(DQB1*0101-*N3) preceded by A(DQB1*0101-*0101) and then by F(DQB1*070101-*070101). Alleles in low ranking genotypes and high ranking genotypes were exclusive to each other and hence represent the allelic substitution as an effect for change in the vaccine response. The variability within DQB1 region of Sirohi goat population was calculated using the Wu-Kabat Variability Index (Table 4).Our result suggest a lot of interesting sites in the amino acid structure of the DQB1, where substitution has taken place. The Wu-Kabat index reveal variability starting from 2.67 at ÃÅ½Ã ²29, ÃÅ½Ã ²60 to 7.19 at ÃÅ½Ã ²81. A total of 19 residues in the translated sequence of DQB1 were found to be polymorphic with at least three amino acid substitutions. Similar results were reported by Amills et al. (2004), where many amino acid residues within and outside the pockets were found to be polymorphic in nature. In present study, although a significant association of these substitutions with vaccine response is not observed, but variability of the region is well explored. Many factors influence the vaccine response as a trait in mammals. Role of environmental factors as well as other MHC and non-MHC genes is important, however apart from that the nature of the responding variable is also one of the most important criteria to look for in such analysis. PPR vaccine is a strong antigen and its invasion produces a cascade of reactions responsible for antibody production. In our earlier study , 94.92% Sirohi kids were observed to be protected with a single dose of PPRV vaccine. Therefore in spite of having variability within the protected category, the differences between the animals is not much and hence association of minor change in the phenotype vis a vis genotype is not visible.Ã There are several studies which revealed the effect of QTLs and non-genetic factors in detail showing the role of non-MHC genes and environmental influences on vaccine response [12,26,27,42,]. In goat, only one study  could show significant association of DRB gene p olymorphism obtained by PCR-RFLP with Johnes disease. Apart from this there are no studies which reveal association of MHC genotypes with disease resistance or susceptibility in goat. 4. Conclusion The genetic variability of DRB and DQB1 gene in Sirohi goat revealed a very rich diversity of this locus with positive evolutionary trend. Our study provide first description of the evidence of such a strong diversity of MHC in Indian goat breed for DRB and DQB region. Due to complex nature of the phenotype, i.e. vaccine response, and good response to the antigen used, association with studied loci was not observed. Apart from this several factors apart from MHC also affected the outcome of the response. Observed variability within the DRB and DQB1 loci reveals potential of the breed for combating several antigenic attacks and hence importance of the studied region in antigen capture and presentation to T cells. Acknowledgements Authors duly acknowledge Department of Biotechnology (GOI) for project grant to carry out the desired work. Authors are thankful to the Director ICAR-CSWRI for providing facilities for carrying out the work. Authors are also thankful to AICRP on Goat for funding the project on Sirohi goat at ICAR-CSWRI Avikanagar. Conflict of Interest Statement: The authors declare that they have no conflict of interest. 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Saturday, January 18, 2020
Shinto in Modern Japan Religion is a constant variable in todayÃ¢â¬â¢s world as well as the past. In order to understand Shinto in modern Japan first Shinto must be looked at from the past. Native Japanese religion states Shinto is the way of the gods. Going into depth of Shinto history and the modern view of Shinto now will bring up where Shinto originated from, itÃ¢â¬â¢s comparison to other religions, and ShintoÃ¢â¬â¢s role in modern Japan. Like many main religions, Shinto originated from prehistoric times but is not truly known because it goes much too far back in time showing as far back as 720 A. D. Its name comes from Chinese words Ã¢â¬Å"shin-taoÃ¢â¬ . Native Japanese religion, Shinto, plays a very significant part in JapanÃ¢â¬â¢s society (Shinto2). Shinto is not only a religion but a way of living for the Japanese. This religion has made a permanent place in history around the world. Writer Chikao Fjisawa pronounced, Ã¢â¬Å"State Shinto Ã¢â¬â a system embodying nationalism loyalty and Emperor Veneration the Shinto was a Ã¢â¬Å"perversion of Shinto theory and beliefs into militaristic and ultra-nationalistic propagandaÃ¢â¬ (Boyd). State Shinto is known as the old Shinto. Just as any other religion, Shinto has been compared to other religions show close resemblance. Shinto has no defined dogma, scared scriptures, or ethical precepts. Japanese tend to combine Shinto with other religious beliefs like Buddhism and Christianity. They also tend not to attach just one of the religious beliefs giving each equal attention. Shrines called jinja is used to practice Shinto and has very distinct gates which make it easy to tell from Buddhist temples. The only reason Shinto was named and systemized in the 16th century was to mark the difference from Buddhism and Confucianism (Shinto1). After World War II a separation between government and Shinto took place. This separation was noted in the constitution and history. As history shows, the emperor issued a statement forbidding use of Shinto symbols as nationalistic reasons and renouncing all rights to divinity. Even in modern day, extremists still favor protests against these and other changes involving the Shinto. Today Shinto is still a strong practice. Many Japanese still use the Shinto shrines for marriage, or to bless a new child, car, and etc. Building, homes and other architectural plots are also known to be blessed for safety and protection. Hundreds of Shinto ceremonies are still carried out daily in todayÃ¢â¬â¢s modern life such as festivals just naming one. In modern life the old Shinto is the main practice still being used. Main religions around the world have an unknown exact birth dates as well as playing an influence role in history. Shinto has not only made its mark in Japanese history, but still withstands in present day society. Religion tends to be a subject who holds controversy and mystery intriguing the minds of almost everyone at some point. ShintoÃ¢â¬â¢s origin may not be able to be definitely pinpointed but itÃ¢â¬â¢s very apparent that Shinto religion still stands very prominent in JapanÃ¢â¬â¢s modern life. As stated earlier, Shinto ceremonies take place daily in Japan and the religion is still practiced by a very large population of Japanese still today. Religion is truly a everlasting time capsule of knowledge telling bits and pieces of the world and relationships within the world. Works Cited Boyd, W. James and Williams, G. Ron Ã¢â¬Å"Reconsidering Shinto after World War II: Perspectives from the Life and Thought of a Shinto Priest. Ã¢â¬ . Motonisa, Yamakage. Ã¢â¬Å"The Essence of Shinto: JapanÃ¢â¬â¢s Spiritual Heart. Ã¢â¬ Kodansha America, Inc. , 2006. Ã¢â¬Å"Shinto. Ã¢â¬ . Ã¢â¬Å"Shinto. Ã¢â¬ .
Friday, January 10, 2020
In the late century, extinction becomes common topic in our daily discussion and debates. An average of 27,000 species is currently extinct each year and there is a possibility of 22% of extinction in the overall species if action is not taken. Animal extinction is due to human immorality and irresponsible behavior just for their own benefits. Since the year 1600, a total of 83 mammals species are known to have become extinct. Wildlife population depleted the trade in live animals, damaged habitats and the countless animals of suffering. Wildlife International, 2008) Apart from that, animal is a good source to make traditional medicine. For example, Chinese believes that snake galls are good to cure diseases and strengthen immune system of the body. Sometimes, human thoughts are powerful which may kill many innocent animals. According to Jeanette McDermott, the book of Bear Muze stated that every year, approximately 10,000 Asiatic black bears are locked in cages the size of their own bodies just to have their bile drained and sold the use in traditional medicine.Besides, many ocean mammals are suffering due to the business benefits. For example, Chinese culture like to consume shark fin soup as part of the cuisine. Businessman will take the advantage to hunt shark fin for money because of its high demand. An organization devoted to preserve marine life, Ocean Environment, stated in Asian Geographic(Nov-Dec 2008) that shark fin soup prized as a delicacy in Chinese cuisine. It is becoming un-cool to consume because 90% of the sharks are un-finned while alive thrown into the sea.