The formation of Salt Traps and their importance in the Gulf of Mexico Research Paper

The formation of Salt Traps and their importance in the Gulf of Mexico - Research Paper Example Whereas many traps can be identified, the salt dome traps will be given the greatest emphasis on this paper. When masses of salt are thrust upward with clastic rocks, they tend to block other substances that might come along porous rocks. Due to the fact that salt is impermeable, they block the hydrocarbons in the underlying permeable rocks and create a reservoir. The Gulf of Mexico is one of the leading petroleum producing provinces in the world for more than 100 years (Hudec, Jackson & Peel 77). It remains one of the most active and intensely explored regions due principally to the tight sand and shale gas which promise greater potentials into the future. The regional stratigraphy of the Gulf of Mexico has been a bit elusive due to the fact that the classification of various features has been generally impeded by the changes of oceanic processes. Nonetheless, deep water margins that occasionally exhibit salt mobility further complicate the potentials of exploration. The most critical aspects of such a program however must rely on adequate understanding of the charge, reservoir development and trap integrity for intensive exploration activities to be deployed (Gemmer et al 202). Charge refers to the time of sedimentation processes as correlated to the allochthonous emplacement of salt as well as the rate of formation of salt welds. The process of reservoir development is critical because it determines the manner of sand distribution during the process of evolution within the basin (Seldon & Flemings 202). The third consideration which is trap integrity is crucial because it is a factor directly related to how salt trap structures chan ge over time and the influence such changes may pose to petroleum migration. The region of the Gulf of Mexico is often cited as the most potential even in the 21st century as a result of the new findings from geological explorations (Fort & Jean-Pierre 270; Seldon & Flemings 202). Deep sea oil exploration traditionally

Ethical issues Article Example | Topics and Well Written Essays - 250 words

Ethical issues - Article Example This is whereby human resource managers face the task of making sure employees are competent to work in the decentralized setting as opposed to how it was before. Employees will have to be able to adapt to the virtual desk to incorporate the concept of flexible work arrangements are incorporated. (Misra, 2009) Another crucial challenged encountered is the global business concept which entails that with the fast growing organizations and the world becoming a global village, human resources managers should be knowledgeable about different cultures traditions and business practices so as to be at per with the globalization process. Thus, a need to keep them always informed on global and international issues. (Misra, 2009) Managing the performance of employees has also been a key area of concern as many human resource managers are not fully aware of all field expectations. Thus, they are incapable of determining performance levels of employees and as a result, fail to access level of competence of employees. Incorporation of all sub systems in human resource management has also proved to be a cog in the wheel. For efficient performance to be achieved, it is vital that the existing departments correlate in order to produce a smooth running department. (Mathis,

Cahi-DRB and DQB1 Alleles in Sirohi Goat

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, 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. [1] 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. [2] 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. [3] 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 [9] 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 [10]. 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) [13]. 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 [14]. 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 [18]. 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 75025â‚ ¬Ã‚ ²E, 26018â‚ ¬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.   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. [3], where DRB.1: 5-TATCCCGTCTCTGCAGCACATTTC-3 and DRB.2: 5-TCGCCGCTGCACACTGAAACTCTC-3 primers were used for amplifying 285 bp product. The reaction mixture of 50ÃŽÂ ¼l comprised of: 10X Taq Buffer (05ÃŽÂ ¼l), 25mM MgCl2 (03ÃŽÂ ¼l), 10mM dNTP (1ÃŽÂ ¼l), 20 pmol (1ÃŽÂ ¼l) of each primer, Taq DNA Polymerase (1IU), Template (1ÃŽÂ ¼l) and Nuclease Free Water (NFW) to make 50ÃŽÂ ¼l. The thermal profile was optimized for amplification of the DRB exon2 as follows: Initial denaturation (94 °C for 4 min), followed by 35 cycles (denaturation for 94 °C for 60 s, annealing at 66 °C for 60s and extension at 72 °C for 60s) and a final extension at 72 °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 [28]. 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 (GenElute„ ¢ 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 [29] 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 [30]. 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. [31], 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 50ÃŽÂ ¼l comprised of: 10X Taq Buffer (05ÃŽÂ ¼l), 25mM MgCl2 (03ÃŽÂ ¼l), 10mM dNTP (1ÃŽÂ ¼l), 20 pmol (1ÃŽÂ ¼l) of each primer, Taq DNA Polymerase (1IU), Template (1ÃŽÂ ¼l) and Nuclease Free Water (NFW) to make 50ÃŽÂ ¼l. The thermal profile was optimized for amplification of the DQB exon2 as follows: Initial denaturation (94 °C for 4 min), followed by 35 cycles (denaturation for 94 °C for 45 s, annealing at 67 °C for 45s and extension at 72 °C for 45s) and a final extension at 72 °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 [28]. 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 [29], 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‚ ¥ 102.5 TCID50 (Raksha-PPR, Indian Immunologicals, India).   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 ˆ’20- ¦C until testing. The ELISA for further analysis was done as described earlier [27]. 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 [32]. 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 [33] with the help of software PAML 4 [34]. The Wu Kabat variability index with respect to amino acids at peptide binding pockets was calculated using the formula given by Wu and Kabat [35]. Index =  Ã‚  Ã‚   The number of different amino acids occurring at a given position  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 [29]. Alleles were Cahi-DRB*0701  Ã‚   (accession no. KX431913), Cahi-DRB*0104  Ã‚   (accession no. KX431914), Cahi-DRB*0402 (accession no. KX431915), Cahi-DRB*0102  Ã‚   (accession no. KX431916), Cahi-DRB*0202  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 [36]. In another study, 11.1 ratio for dN/dS was recorded in Peptide Binding Region of 12 Chinese indigenous goats for DRB*02 sequences [6]. PBR being polymorphic, its importance is seen here. According to Simmons et al. [37], 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 [38], 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 [30]. 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. [9] reported a ratio of 2.14 in Nigerian goat breeds.   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.   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 69.99 ±0.42 (Min 13.32, max 91.60) with minimum PI 35.12 and maximum PI 98.82.  Ã‚   Average   age   at vaccination   was   142.43    ±Ã‚   14.67   days   with   minimum   age   93 days   and maximum   age   164   days.   Variability in the vaccine response was evident in the lambs.   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).   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 [39]. 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 ÃŽÂ ²65 (6.41%) and ÃŽÂ ²73 (5.94%).   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 ÃŽÂ ²29, ÃŽÂ ²60 to 7.19 at ÃŽÂ 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 [27], 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.   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 [8] 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. References Herrmann-Hoesing LM, White SN, Mousel MR, Lewis GS, Knowles DP (2008) Ovine progressive pneumonia provirus levels associate with breed and Ovar DRB1. Immunogenet 60:749-758 Larruskain A, Minguijà ³n E, Garcà ­a-Etxebarria K, Moreno B, Arostegui I, Juste RA, Jugo BM (2010) MHC class II DRB1 gene polymorphism in the pathogenesis of Maedi-Visna and pulmonary adenocarcinoma viral diseases in sheep. Immunogenet 62:75-83 Amills M, Francino O (1995) Nested PCR allows the characterization of Taq I and Pst I RFLPs in the second exon of the Caprine MHC class II DRB gene. Vet Immunol Immunopathol 48:313-321 Amills M, Francino O (1996) A PCR-RFLP typing method for the Caprine MHC class II DRB gene. Vet Immunol Immunopathol 55:255-260 Dongxiao S, Yuan Z (2004) Polymorphisms of the second exon of MHC-DRB gene in Chinese local sheep and goat. 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Children Are Influenced by T.V. :: Media Argumentative Persuasive Argument

Children Are Influenced by T.V.      Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚   When children are young, they tend to act different than when they are older. They are not mature yet and are still easily influenced.   Little kids are usually very easily influenced by their surroundings.   Whether it is television, friends, family members, or just plain strangers, everyone and everything are influences on a little kid.   I believe that young boys are usually more easily influenced than young girls.      Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  First, one influence that has a major effect on children's behavior is television.   Believe it or not, T.V. plays a big role in how kids act. Shows like the Power Rangers and Beetleborgs can make boys violent and want to fight.   Girls are not so easily influenced by television as boys are. Although there are some girls that want to fight along with the boys, for the most part, girls do not like those types of shows.      Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚   Along with television, children may also be influenced by people they don't know.   Although most children are taught not to talk to strangers, we would be surprised how many actually do.   Studies have shown that both boys and girls do talk to strangers, but boys are more likely to do what a stranger tells them than girls are.   A lot of young boys are easily deceived.   Girls, on the other hand, are more cautious.   Even when girls are young, they are still weary of trusting people they don't know. There are, of course, exceptions to every rule.      Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚   Finally, kids are most influenced by their parents and their friends. Because friends and family members are people that kids trust, they tend to want to be like them.   Parents play a big role in a child's life.   If a kid sees his/her parents fight all the time at home, it will probably make the child violent or secluded.   If a kid hears his friends cuss all the time, then he/she will probably start cussing too.   If a child, whether it be a boy or a girl, places a lot of trust in a person, every

Hamlet’s Other Apparition Essay

I was lying in on a hard cold floor. My head was reeling with a thousand bright lights – stars everywhere. Is this another apparition? The ghost of my father beckons me? I realized the cold floor along my back. The incessant throbbing in my head echoes a swosh, swosh sound ringing in my ear. The cold permeates me, my body is chilled but it does not move. The somewhat forgotten heaviness of responsibility returned and settled like a warm heavy cloak. Ah! what relief to shake it off. I lie there with the wet cold soaking through the wool and velvet of my coat and clothes to me and in me. I was wrong; there were no bright lights, only darkness. The sound receded to the distance, the throbbing in my head traveled to my temples and stayed there, as if to remind me that this is no apparition, as I had hoped it was, setting me free from this cursed mission to set things right. The knowledge surprised me into sitting up. The cold retreated somewhat. I knew now the source of the chill. It was the stone floor damp by the winter night’s mist. The dark did not retreat as everything else had – the pain in my head, the ringing in my ear and the cold. It was still dark – blindingly so. Suddenly, there appears a glow of light in front of me like a white dot from afar, visible only because of the total darkness. Ah, the apparition. It reminded me of a star in a dark, moonless night. Only it was lonesome, like I usually am standing in the tallest turret of the castle watching the heavens. Only now, I am not in the turret warmly surrounded by a million stars of the heaven, I am in this long dank corridor guided by a lonesome star. I felt around the floor for what I do not know. I did not summon the servants, for some reason I sat there in the damp stone floor feeling for something. No such luck in the dark. The light was merely a white dot and not a confirmation of a long held suspicion. I became aware of a flowing wetness in my neck, leaking from my ear. I touched it. It was sticky. Blood? Mine? Where was I? I feel that I should be wearing something in my head – my, mind, perhaps? I felt around some more, forgetting myself and crawling around in the dark like an animal. A poor animal not gifted with night vision. I stopped. Am I now mad in fact and not just in act? I stood up. The sudden movement sent me spinning. Was there an abyss in the castle? Why am I falling and spinning out of control. The bright lights threatened to come again. I closed my eyes fiercely. My head was pounding now. The swosh, swosh ringing in my ear threatened to return. I feel like I would embarrass myself as I keeled forward. Is it possible to embarrass oneself while alone in the dark? But what of embarrassment, have I not met with some maiden unkempt and uncouth to breathe deeply in her ear? Ah, madness, is it you? With closed eyes, I tried to stay still. The abyss, the pounding, and the ringing receded. They still threatened from a close distance but at least they were bearable and I was able to stay on my feet. Gingerly, I took a step forward, towards that white dot in the distance. The nausea came back but I conquered it. I took another step, and barely stayed on my feet. I raised my hand from my side and found a wall. Finally, support for my unfamiliar body. I made baby steps toward the bright light. I heard sounds. Is the ringing in my ear coming back to hunt me? No, it was different. It was like the rumblings of the sea from the distance. Am I still in Elsinore? As I draw nearer to light the sounds became the roar of waves. I remember the crash of waves in the moors. I could almost taste the salt in the air. I anticipate the fresh salty smell of the sea. It is what I need. Suddenly, I feel as if the hallway was suffocating me. The dark was closing near. I tried to make my way to light faster. My steps were small, but hurried, propelled by need to breathe in the sea. A soft breeze came to me. Ah! The sea, it beckons temptingly. The light became bigger. It became a slash, instead of a dot. It was long standing white line in the distance. The crash of waves became rumbles. The sea was gone even if the cool breeze remains, becomes more constant. The rumbles became murmurs. The words rush and tumble with each other, like a hurriedly spoken prayer, a long one from the sound of it. Little by little I hear a voice†¦ The light†¦ The light†¦ It was gone and a barely open door stands in its place. What is wrong with me? The whimsy of it all came back to taunt me †¦ my father’s ghost†¦ stars†¦ abyss†¦sea†¦ a guiding light†¦ Must kill! Must Kill! Was that me? No, it was the voice, a long familiar voice. I know that voice. I know that†¦ I did not rush to open the door. I peeped inside, trying to fit my vision in that long white line the door allowed. There was movement from inside. Then, before me is a face, a familiar face, familial face. I know that face. I know that face. You! Then, in came the abyss. There was dark and then, light – extremely bright light. Ah, the sun. It warms me, it bathes me. It seeps through the covers, to my night shirt, to my body. My frail mother, shifting sheets again? Ophelia? I was lying in a wonderfully warm bed and covered by wonderfully warm sheets. Where has the winter gone? The corridor! The room! The face! Where had they all gone? My head was reeling with a thousand questions. The light of day does not illuminate the dark corners of my suspicion. I pray for the throbbing in my head, the ringing in my ear. For then, there will be no questions, only answers and – vengeance! The doubts forced me into sitting up. The warmth retreated somewhat. The cool winter morning penetrates the sheets and touched my skin bringing with it wakefulness and resolve. I touched my ear and found something sticky. Slowly, an eerie smirk made it way into my face. It must be so†¦ It must be so†¦

Corporation information Essay

This section it’s dedicated to provide all the information related to the company that our clients need, and to offer a brief history of how the company was created as a responsible company and providing a service that exceeds our customer’s expectations. Our company recognizes that accomplishment and maintenance of security program is our personal responsibility, therefore, we will have to take initiative and be an example, this way we will maintain our system implemented by different techniques and tools like: Trainings on C-TPAT Security processes described * Security Controled and Registred. * Technology of innovation * Personnel recruited * Internal Audits * Selection of business partners Since we look for  implementation and development of the program of security C-TPAT we must fulfill some requirements to maintain a greater security in all the chain of supplies. Thus we have verifiable writings for selection of our businesses partners. We request for procedures of security and the processes to fulfill the minimum requirements of security established by the C-TPAT, participating in a questionnaire applied by our company. When completing this questionnaire, not only will help us to fulfill our obligations like member of the C-TPAT, but also some of our customers will look for fulfill requirements for importer and to become members of the C-TPAT. Guarantee security of our processes and our clients’ also. Questionnaire is designed to identify those areas where improvements are necessary to fulfill minimm security requirements. Correct identification of a security weakness will not affect our relation of businesses, but it will allow us to work with our clients to develop a plan of security improvement if necessary. All the people working on the company contribute to our success. An integration of our different talents and perspective, stimulate new and creative opportunities for our business. Collectively, it will generate a rewarding and more exciting work atmosphere where each individual feels like person in charge of performance of our company. We respect rights and dignity of all employees. We made an effort for being a company that independently attracts the best people, no matter origin, beliefs or life style. We are commited to create a work atmosphere with confidence and respect, inclusion and diversity, and employees are listened to improve team work. We assure all employees know our job enviroment, tasks, responsibilities and general activities to aim our organization. We develop their capacities through open and constructive conversations on their professional direction. Also we try to guarantee that all the employees are recognized and compensated by their yield. For our company the responsibility of each individual as employe is being conscious of policies and be responsible of his own behavior. Our integrity and reputation as company will not have to be committed. Any attitude that can seem questionable is not acceptable. (Customs Trade Partnership Against Terrorism), is a joint initiative of the government and businesses of create cooperation relations to fortify security of supply chains and borders. By this project, customs requests all companies to guarantee integrity of their security and to communicate this to commercial partners of supply chain. The company look for a certification as member of C-TPAT. Obtaining this certification, we will offer a better service to our customers with great security, reliance and rapidity exporting cargo, also our customers (importers) and our company will be benefitted with results obtained with implementation of C-TPAT. In order to articipate in C-TPAT, companies must elaborate and apply programs of integral security to improve security internal and external with partners and clients. Therefore, we have developed and maintained a complete and detailed security plan with directives of values of EE. UU. Customs. As C-TPAT includes all components of supply chain, such as importers, carriers, intermediaries, operators of warehouses and manufacturers, the company is determined to communicate security requirements for C-TPAT to suppliers and clie nts.