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Veterinary Infectious Diseases
Ryan Arsenault
University of Delaware, United States
Michael Kogut
Agricultural Research Service, United States Department of Agriculture, United States
Guillermo Tellez
University of Arkansas, United States
Application of Monoclonal Antibodies Developed Against the IpaJ Protein for Detection of Chickens Infected With Salmonella enterica Serovar Pullorum Using Competitive ELISA
Kequan Yin1,2,3†, Jingwei Ren1,2,3†, Yue Zhu1,2,3†, Lijuan Xu1,2,3, Chao Yin1,2,3, Yang Li1,2,3, Yu Yuan1,2,3, Qiuchun Li1,2,3* and Xinan Jiao1,2,3
1Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agri-food Safety and Quality, Ministry of Agriculture of China, Yangzhou University, Yangzhou, China
2Jiangsu Key Lab of Zoonosis/Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, China
3Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China
Pullorum disease remains an epidemic in the poultry industry in China. The causing pathogen is a host-restricted Salmonella enterica serovar Pullorum, which can spread through both horizontal and vertical transmissions. To eradicate the pullorum disease from poultry farms, it is necessary to specifically monitor the prevalence of the bacterial infection in adult chicks. In this study, we constructed a new competitive ELISA method based on the development of monoclonal antibodies (MAbs) against a specific immunogen of S. Pullorum, IpaJ protein. In total, eight MAbs against IpaJ were prepared using the purified recombinant His-IpaJ protein as the immunogen. Characterization of the eight MAbs demonstrated that 4G5 can be used as the competitive antibody in ELISA. A competitive ELISA was subsequently developed using purified MBP-IpaJ as the capture (0.5 μg/ml) and the HRP-labeled 4G5 (0.14 μg/ml) as the competitive antibody, respectively. A specificity test demonstrated that the ELISA assay can differentiate antisera of S. Pullorum-infected chickens from that of S. Gallinarum and S. Enteritidis. Furthermore, 4 out of 200 clinical antisera collected from a poultry farm were detected to be S. Pulloram positive using this method. The plate agglutination test (PAT) and the previously established indirect ELISA confirmed that these positive antisera reacted specifically with S. Pullorum. We propose that the established competitive ELISA assay based on MAb against IpaJ protein, is a novel and quick method that can detect S. Pullroum infection in the poultry industry.
Avian salmonellosis remains a big threat to the poultry industry and the public health system. The pullorum disease has mostly been eradicated in many developed countries in Europe, America, and Asia, apart from sporadic incidences in free-ranging backyards. However, it remains endemic in some developing countries like China (1). Monitoring the prevalence of pullorum diseases or adult chickens harboring the pathogen (Salmonella enterica serovar Pullorum, S. Pullorum) in poultry farms is necessary to control the transmission of the bacteria for the eradication of the disease. However, the traditional method to diagnose the disease is to identify the pathogen in infected chickens through biochemical tests or molecular detection methods including PCR analysis (2–4). All of these methods need the bacteria to be extracted from the chicken samples, which is time-consuming. Additionally, the adult birds which are the carrier of S. Pullorum may continue to shed Salmonella in eggs because of its colonization in the reproductive tract, but the pathogen cannot be detected in the intestine, liver, and spleen. Thus, new methods based on the detection of antibodies can improve the efficiency of pathogen detection, as opposed to the traditional methods (5).
Previous studies have developed many serological detection methods to identify the Salmonella infection in human or animals. Most of these studies used cell surface proteins as the detection antigens, such as LPS, FliC, and so on (5–8). However, the specificity of these molecules is not able to differentiate the various Salmonella serotypes. For example, the plate agglutination test (PAT) based on O9 antigens could not differentiate S. Pullorum from S. Gallinarum, and S. Enteritidis (1). To establish a serological detection method specific to S. Pullorum infection, an antigen specific to S. Pullorum should be found and evaluated against the antibodies produced during the bacterial infection. Our previous studies demonstrated that IpaJ protein was prevalent in 99.08% of S. Pullorum and was deficient in other Salmonella serotypes, including the closely related S. Gallinarum and S. Enteritidis species. Additionally, the antibodies against IpaJ protein could be detected from the first to the twelfth week after S. Pullorum infection in chicken sera (9).
Based on our previous established indirect ELISA assay to detect antibodies against IpaJ protein in chicken antisera, we tried to establish a new and efficient method to monitor S. Pullorum infection in poultry industry. In the present study, we prepared monoclonal antibodies (MAbs) against IpaJ and developed a competitive ELISA method to detect S. Pullorum infection in poultry farms with one of the MAbs.
Bacterial Strains, Plasmids, and Proteins
S. Pullorum reference strain C79-13 was offered by China Institute of Veterinary Drug Control. The prokaryotic expression vector pColdI (Takara, Japan) was used to construct recombinant expression plasmids in host cells E. coli BL21(DE3). The bacteria with recombinant plasmids were grown in Luria Bertani (LB) broth with Ampicillin (100 μg/ml). The purified MBP-IpaJ was collected and preserved in our laboratory (9).
Construction of Recombinant Expression Plasmid
According to the published ipaJ gene sequence (Accession Number: GU949535) in GenBank, forward and reverse primers pColdI-ipaJ-F/R (coldI-ipaJ-F: CATCATCATCATCATCATATGCGGTTAAAATTTATCAG; coldI-ipaJ-R: TCTAGACTGCAGGTCGACTCAAGCTGACAAGACAATAGA) were designed to clone the gene from C79-13 using PCR. PCR analysis was performed in a 50 μ reaction system: 2.5 U of Taq DNA polymerase, 10 μ 5 × SF buffer, 200 μM dNTP mix, 0.4 μM upstream and downstream primers, and 1 μl of DNA template. The amplification procedure was started with a pre-denaturation at 95°C for 5 min, followed by 30 cycles of 95°C for 50 s, 61°C for 50 s, and 72°C for 1 min, and terminated with a final extension at 72°C for 10 min. Next, the PCR products were resolved on a 1% agarose gel and purified using MiniBEST Agarose Gel DNA Extraction kit (Takara, Japan). The purified PCR products of ipaJ were then ligated to pColdI vector and transformed into the competent E. coli DH5α. The positive colonies carrying the recombinant plasmid were identified using PCR and sequencing analysis. The recombinant plasmids pColdI-ipaJ were then transformed into E. coli BL21(DE3) competent cells to produce BL21(DE3)-pColdI-ipaJ.
Expression and Purification of the Recombinant His-IpaJ
An overnight culture of recombinant bacteria BL21(DE3)-pColdI-ipaJ was inoculated into fresh LB medium with ampicillin at 1:100 dilution. When the OD600 was between 0.4 and 0.6, the inducer IPTG was added to the medium with the final concentration of 0.5 mM to induce protein expression. The bacteria were then cultured at 15°C for 24 h with continuous shaking at 150–180 rpm. The bacterial pellets were collected for ultrasonic lysis, and the precipitate including His-IpaJ protein as inclusive body was subjected to SDS-PAGE followed by purification from the gel according to the protocol of "Purification proteins from polyacrylamide gels (TECH TIP #51, Thermo scientific, USA)" with a few modifications. Briefly, gel was stained with 1 M KCl for 3–5 min, the white stained protein band of interest was excised and cut into pieces by a clean pestle. The excised gel pieces were kept in a clean microcentrifuge tube and 0.5–1 ml of elution buffer (50 mM Tris-HCl, 150 mM NaCl, and 0.1 mM EDTA; pH7.5) was added to completely immerse the gel pieces; After overnight incubation at 30°C in a rotary shaker, the tube was centrifuged at 5,000–10,000 × g for 10 min and the supernatant was pipetted into a new microcentrifuge tube. The supernatant was tested by SDS-PAGE for purified protein, and the concentration of the protein was monitored by Quick StartTM Bradford protein assay (Bio-rad, USA).
Immunization of Mice With His-IpaJ
To prepare the B cells producing antibodies against IpaJ protein, 6–8 weeks old BALB/c mice were immunized with His-IpaJ protein (100 μg per mouse) by intraperitoneal (i.p.) injection. The immunization was performed twice every 2 weeks. Three days prior to fusion, 100 μg of His-IpaJ was intravenously injected into the mice. The spleen cells were then fused with sp2/0 cells by using the lymphocyte hybridoma technique (10). All of the animal experiments and managements were undertaken by the permission of the Animal Welfare and Ethics Committees of Yangzhou University, and complied with the guidelines of the institutional administrative committee and ethics committee of laboratory animals.
Preparation of Monoclonal Antibodies Against IpaJ
The positive hybridoma clones expressing antibodies against His-IpaJ were screened using the previously established indirect ELISA method, with MBP-IpaJ recombinant protein as the coating antigen (9). Positive clones were sub-cloned three times by the limiting dilution method. The Ig sub-class of MAbs were identified by using a mouse mAb isotyping kit (Sigma, USA) according to the manufacturer's instruction. The positive hybridoma cell lines secreting anti-IpaJ MAbs were injected intraperitoneally into BALB/c mice to grow and proliferate. Ascites fluids containing abundant anti-IpaJ MAbs were collected from the immunized mice and purified by protein A chromatography (GE Healthcare). The purified MAbs were send to GenScript Biotechnique Company (Nanjing, China) for biotinylation with HRP.
The cell lysates or purified recombinant proteins were subjected to SDS-PAGE on a 12% polyacrylamide gel in Tris-glycine running buffer (pH 8.7), and transferred to PVDF membrane (Pall, USA) by PyxisTM Gel Processor (Pyxis, China). The PVDF membrane was blocked in 5% BSA and then incubated with anti-MBP antibody or anti-IpaJ MAbs in blocking buffer for 2 h at 37°C. After washing, the membranes were incubated with goat anti-mouse IgG-HRP (Sigma, USA) for 1 h. The PVDF membrane was then stained with the ECL chromogenic kit (Thermo, USA) and scanned using the Amersham Imager 600 imagers (GE Healthcare, USA).
Construction of a Competitive ELISA Assay
To determine the best working concentration of the coating antigen and the MAb in the competitive ELISA assay, 96-well plates were coated with the purified protein MBP-IpaJ at the following concentrations: 4, 2, 1, 0.5, and 0.25 μg/ml. Twelve wells per concentration were coated with 100 μl per well at 4°C for 14–16 h. Subsequently, the plates were washed three times and then blocked with PBS containing 2% BSA in PBS at 37°C for 2 h. After washing, 50 μl of the doubling diluted HPR-labeled MAb from 1:1,000 to 1:64,000 and 50 μl of the doubling diluted S. Pullorum positive antiserum from 1:10 to 1:10,240 were mixed and added into each cell and incubated for 2 h at 37°C. The wells were then washed as above. HPR activity was revealed by adding 3,3′, 5,5′-Tetramethylbenzidine (TMB) at 37°C and incubating for 10 min, and then stopped by adding 2 M H2SO4. The OD450 was measured to determine the best concentration of the MBP-IpaJ and the MAb used in the competitive ELISA.
Based on the working concentration of the coating protein and the MAb, the dilution ratio of the detected antiserum was determined as follows: 50 μl of the S. Pullorum positive antiserum was double diluted from 1:2 to 1:128 and 50 μl of the HRP-labeled MAb was added into the ELISA plate coated with the MBP-IpaJ. The OD450 was measured to calculate the inhibition rate of the antiserum to the MAb.
Determination of Cut-Off Value for the Competitive ELISA Assay
The specificity and sensitivity of the competitive ELISA assay were checked using other antisera from chickens infected with S. Pullorum or other avian Salmonella serotypes such as S. Gallinarum, S. Enteritidis, S. Typhimurium, and other poultry bacteria, including E. coli, and Shigella flexneri. The antisera from chickens infected with these bacteria were subjected to the competitive ELISA assay to evaluate the specificity and determine the cut-off value on the basis of the average control samples. The Gradprism 6.0 software was used to analyze the data and produce ROC curve for determination of the cut-off value.
Application of the Competitive ELISA Assay to Clinical Samples
In a poultry farm, 200 clinical antiserum samples were collected from chickens. The samples were preserved at −70°C and subsequently monitored using the established competitive ELISA assay. An assay for each sample was performed in triplicates, and the detected positive samples were confirmed by PAT and the indirect ELISA assay (9).
Expression and Purification of Recombination Protein His-IpaJ
The ipaJ gene of 840 bp was successfully amplified from C79-13 and cloned into the prokaryotic vector pColdI, to produce pColdI-ipaJ. The sequence of ipaJ was identified using sequencing analysis and digestion with restriction enzymes (Figure 1A).
Figure 1. Expression, purification, and identification of recombinant expressed protein His-IpaJ. (A) Construction of the prokaryotic recombinant plasmid pColdI-ipaJ. Lane 1,2: plasmid digested with NdeI and SalI (Takara, Japan). (B) SDS-PAGE analysis of expressed recombinant protein His-IpaJ. Lane 1: induced BL21(DE3)-pColdI as negative control; non-induced BL21(DE3)-pColdI-ipaJ. Lane 3: the supernatants of induced BL21(DE3)-pColdI-ipaJ. Lane 4: the precipitants of induced BL21(DE3)-pColdI-ipaJ. (C) Purified recombinant His-IpaJ protein (32 KDa). (D) Western-blot analysis of the recombinant His-IpaJ protein, using antibody against His Tag.
The recombinant bacteria BL21(DE3)-pColdI-ipaJ was induced by 0.5 mM IPTG, and the SDS-PAGE results confirmed that His-IpaJ was successfully expressed in the host cells (Figure 1B). The recombinant protein was estimated to be approximately 32 KDa in the form of inclusion body in BL21(DE3). The His-IpaJ protein was then purified from polyacrylamide gels (Figure 1C).
The His-IpaJ protein expressed and purified from BL21(DE3) was further identified by Western blot analysis with anti-His antibody. The results showed that the expressed recombinant protein His-IpaJ could specifically react with anti-His antibody (Figure 1D).
Generation and Characterization of MAbs Against His-IpaJ
After two independent fusions and three sub-clonings, eight hybridomas (1C3, 1F3, 2D9, 3B5, 3D8, 4D5, and 5D1) were obtained and selected for further studies based on their strong reactivity with MBP-IpaJ. The results of IgG isotypes showed that four MAbs (1C3, 2D9, 4G6, and 5D1) were IgG1, two MAbs (1F3 and 3D8) were IgG2b and two MAbs (3B5 and 4D5) were IgM (Table 1). The titers of culture supernatants and ascites of each MAb revealed that 4G6 and 5D1 were better than the other MAbs (Table 1). Western blot analysis confirmed that all of the eight MAbs could react specifically with the His-IpaJ and the MBP-IpaJ (Figure 2). Additionally, seven MAbs showed strong reaction with IpaJ secreted by S. Pullorum cultured in LB medium by Western blot analysis (Figure 3), but did not react with the lysates of other Salmonella serotypes or E. coli. However, the Western blot analysis revealed that these MAbs also reacted with supernatants of Shigella flexneria, since Shigella flexneria expressed a similar IpaJ protein, with 49% homology to the IpaJ in S. Pullorum.
Table 1. Characteristics of eight monoclonal antibodies against IpaJ.
Figure 2. Identification of the reactive specificity of monoclonal antibodies against IpaJ protein by Western blot. Lane 1: Purified His-IpaJ protein (32 KDa); Lane 2: the supernatants of induced BL21(DE3)-pColdI; Lane 3: Purified MBP-IpaJ protein (71 KDa); Lane 4: the supernatants of induced BL21(DE3)-pMAL-c5X. Lane 5: Purified MBP-IpaJ (from Shigella flexneri) protein. The 1C3, 1F3, 2D5, 3B5, 3D8, 4D5, 4G6, and 5D1 represents different monoclonal hybridoma cell lines.
Figure 3. The reactive specificity of monoclonal antibodies against proteins expressed by different Salmonella serotypes. The supernatants of different Salmonella serotypes were used as the antigens. S. Pullorum C79-13 strain was used as a positive control (Lane 1); S. Gallinarum Sg9 (Lane 2); S. Enteritidis P125109 (Lane 3); S. Dublin D2 (Lane 4); S. Typhimurium SL1344 (Lane 5); S. Derby LQSD15 (Lane 6); S. Indiana LQSI12 (Lane 7); Escherichia coli BL21(DE3) was used as a negative control (Lane 8); Shigella flexneri SNI carrying IpaJ was used as another positive control.
The Establishment of a Competitive ELISA Assay for S. Pullorum Infection
Our previous study showed that chickens could produce antibodies against IpaJ protein 1 week after infection with S. Pullorum and kept at a stable level for up to 12 weeks (9). In order to detect S. Pullorum infection, a competitive ELISA was developed using purified MBP-IpaJ as capture and HRP-labeled MAb 4G5 as detection antibodies, respectively. The checkerboard analysis revealed that the appropriate concentration of the coating antigen was 0.5 μg/ml, and the working dilution of the HRP-4G5 was 0.14 μg/ml (Table 2). The inhibition rate detected by positive antisera from S. Pullorum infected chickens demonstrated that the working dilution ratio of antisera was 1:16 (Supplementary Figure 1). The cross-link point between sensitivity and specificity curves reflected that the cut-off for inhibition rate was 40.5, and the Youden index was 0.668 (Figure 4). This demonstrated that when the inhibition rate was ≥40.5, the antiserum samples were considered positive for antibodies against IpaJ, and vice versa.
Table 2. Checker-board analysis for concentration of coating protein (MBP-IpaJ) and antibody (HRP-4G6) in the competitive ELISA method.
Figure 4. Determination of the cut-off value and Youden index of the competitive ELISA method. Thirty two positive antisera from S. Pullorum-infected chickens and 25 negative antisera from SPF chickens not exposed to Salmonella were subjected to the ELISA assay to obtain the cut-off value (A) and the Youden index (B). The cut-off value of 40.5 represents when the inhibition rate of antisera samples should be ≥40.5.
Application of the Competitive ELISA Method to Monitor Clinical Samples
The competitive ELISA method was used to detect 200 clinical antiserum samples from a poultry farm, and the result showed that four samples were positive for antibodies against IpaJ. The PAT method and the indirect ELISA assay further confirmed that all of the four antiserum samples could react with S. Pullorum C79-13. However, the PAT cannot discriminate antisera from chickens infected by S. Pullorum, S. Gallinarum, or S. Enteritidis with the same O9 antigen, and the indirect ELISA method is time-consuming compared to the competitive ELISA assay.
S. Pullorum and the other avian pathogens, S. Enteritidis and S. Gallinarum, are group D members in the well-recognized Kauffmann-White Salmonella classification scheme (11). Since these serotypes have similar LPS O-antigen structure, the established ELISA methods based on the anti-LPS MAbs can be used to monitor avian salmonellosis caused by S. Pullorum, S. Gallinarum, S. Enteritidis, and other Salmonella group D serotypes (12). These methods can be applied for the surveillance of avian salmonellosis in the poultry farms, but cannot identify the exact serotype of the pathogen. In order to differentiate S. Pullorum-infection from other serotype infections, we established a competitive ELISA method to specifically detect S. Pullorum-infected chickens.
IpaJ has been identified as an immunogen, expressed specifically by >99% of S. Pullorum isolates, and not detected in other closely related group D serotypes including S. Gallinarum, and S. Enteritidis (4). Our previous study demonstrated that the protein could induce antibody production at 1 week post-infection and persist for over 12 weeks (9). An indirect ELISA method based on the MBP-IpaJ recombinant protein as the coating protein was established and applied for detection of antibodies from S. Pullorum-infected chickens (9). However, since the IpaJ protein has the MBP tag, the indirect ELISA method has low specificity. Therefore, the MAbs against IpaJ were obtained and the competitive ELISA method was constructed and applied to the clinical samples in this study. The antiserum samples and the HRP-labeled MAbs were added at the same time in the competitive ELISA assay, which saved more time than the indirect ELISA method. In S. Pullorum, the IpaJ works as an immune regulator involved in inhibiting T-cell immune response (13). However, the molecular mechanism behind its immune modulatory role remains unknown. Thus, the MAbs against the IpaJ identified in this study may also be used as a tool to further explore its function.
We conclude, that the present study obtained eight MAbs against the IpaJ of S. Pullorum, and developed a competitive ELISA method for identifying S. Pullorum infection in poultry farms based on the MAb 4G5. The method showed enhanced sensitivity and specificity compared to our previously established indirect ELISA method. We thus propose that this competitive ELISA is a quick and efficient method that can be applied for the surveillance of S. Pullorum infection in the poultry industry.
All datasets generated for this study are included in the article/Supplementary Material.
The animal study was reviewed and approved by the Animal Welfare and Ethics Committees of Yangzhou University.
KY, QL, and XJ designed and coordinated the study. KY, JR, and YZ performed the experiments and wrote the manuscript. KY, LX, CY, YL, and YY analyzed the data. QL and XJ reviewed and edited the manuscript.
This study was supported by the National Key Research and Development Program of China (2017YFD0500705; 2017YFD0500100); Special Fund for Agro-scientific Research in the Public Interest (201403054); National Natural Science Foundation of China (31730094, 31320103907); Jiangsu province agricultural science and technology independent innovation funds (CX(16)1028); The Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).
The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fvets.2019.00386/full#supplementary-material
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Keywords: Salmonella enterica serovar Pullorum (S. Pullorum), monoclonal antibody (MAb), competitive ELISA, IpaJ, plate agglutination test (PAT)
Citation: Yin K, Ren J, Zhu Y, Xu L, Yin C, Li Y, Yuan Y, Li Q and Jiao X (2019) Application of Monoclonal Antibodies Developed Against the IpaJ Protein for Detection of Chickens Infected With Salmonella enterica Serovar Pullorum Using Competitive ELISA. Front. Vet. Sci. 6:386. doi: 10.3389/fvets.2019.00386
Ryan Arsenault, University of Delaware, United States
Guillermo Tellez, University of Arkansas, United States
Michael Kogut, United States Department of Agriculture, United States
Copyright © 2019 Yin, Ren, Zhu, Xu, Yin, Li, Yuan, Li and Jiao. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Qiuchun Li, qcli@yzu.edu.cn
†These authors have contributed equally to this work


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Chief Justice Anin-Yeboah is the first male chief justice after 12 years of female appointees. He is a man with enormous experience as far as Ghana's legal system is concerned. Justice Anin-Yeboah was born in May 1953 in Koforidua, Eastern Region but belongs to the Toase village in the Ashanti Region. He attended Amaniampong Secondary School and Apam Secondary School. He studied law at University of Ghana and went on to graduate from the Ghana School of law in 1981.
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We congratulate Chief Justice Anin-Yeboah and wish him the very best in his continuous service to our country and the highest law courts. God bless our supreme court and God bless mother Ghana.
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The C-O-P bond 3'' to the apurinic or apyrimidinic site in DNA is broken by a beta-elimination reaction, leaving a 3''-terminal unsaturated sugar and a product with a terminal 5''-phosphate. The C-O-P bond 3'' to the apurinic or apyrimidinic site in DNA is broken by a beta-elimination reaction, leaving a 3''-terminal unsaturated sugar and a product with a terminal 5''-phosphate.
Pa-AGOG, the founding member of a new family of archaeal 8-oxoguanine DNA-glycosylases.
Structures of end products resulting from lesion processing by a DNA glycosylase/lyase.
A DNA glycosylase from Pyrobaculum aerophilum with an 8-oxoguanine binding mode and a noncanonical helix-hairpin-helix structure.
E.C. 3.1.25.2 was transferred to E.C. 4.2.99.18.
This enzyme is homologous to hOgg1 (PDB; 1m3h, 1m3q), and probably catalyzes similar reactions to those by hOgg1. The literature  suggests the catalytic mechanism of hOgg1.
However, the detailed mechanism has not been elucidated yet.


text:
Marquette.edu // Raynor Memorial Libraries // Archives // Nativeguide //
GUIDE TO CATHOLIC RECORDS ABOUT NATIVE AMERICANS IN THE U.S.
Volume 3: Western United States
University of Texas at Arlington. Special Collections, TX-1
Texas Tech University. Southwest Collection and Special Collections Library, TX-28
Catholic Church. Catholic Archives of Texas, TX-2
St. John Bosco Church, TX-29
Texas State Library and Archives Commission. Texas State Archives, TX-3
University of Texas at Austin. Alexander Architectural Archive, TX-4
Bexar County Courthouse. Spanish Archives Department, TX-30
University of Texas at Austin. Benson Latin American Collection, TX-5
Catholic Church. Archdiocese of San Antonio, TX-31
University of Texas at Austin. Research and Collections Division, TX-6
Central Catholic High School, TX-32
University of Texas at Austin. Harry Ransom Humanities Research Center, TX-7
Daughters of the Republic of Texas. Library, TX-33
Holy Cross High School, TX-34
Benedictine Sisters. St. Scholastica's Monastery, TX-8
Holy Spirit Church, TX-35
Incarnate Word High School, TX-36
Antonian College Preparatory High School, TX-9
St. Anthony de Padua Church, TX-37
St. Anthony High School, TX-38
Texas A & M University. Cushing Memorial Library and Archives, TX-10
St. Gregory the Great Church, TX-39
St. John Berchmans Church, TX-40
St. Monica Church, TX-11
St. Luke Church, TX-41
St. Margaret Mary Church, TX-42
Corpus Christi Public Libraries. Special Collections and Archives, TX-12
St. Mary Church, TX-43
Society of Our Lady of the Most Holy Trinity, TX-13
St. Mary Magdalen Church, TX-44
St. Mary's University. TX-45
Catholic Church. Diocese of Dallas. Archives and Museum, TX-14
St. Peter, Prince of the Apostles Church, TX-46
Dallas Historical Society. G.B. Dealey Library, TX-15
St. Pius X Church, TX-47
Dallas Public Library. Dallas History and Archives, TX-16
San Antonio Public Library. Texana and Genealogy Department, TX-48
San Fernando Cathedral, TX-49
University of North Texas. Rare Book and Texana Collections, TX-17
San Francisco de la Espada Mission, TX-50
San Jose y San Miguel Mission, TX-51
Our Lady of Refuge Church, TX-18
San Juan Capistrano Mission, TX-52
Sisters of Charity of the Incarnate Word, TX-53
Catholic Church. Diocese of El Paso. Chancery, TX-19
U.S. National Park Service. San Antonio Missions National Historic Park. Headquarters, TX-54
El Paso County Historical Society. Jane Burges Perrenot Research Library, TX-20
El Paso Public Library. Border Heritage Center, TX-21
Institute of Texan Cultures Library, TX-55
Our Lady of Mount Carmel Church and School, TX-22
Special Collections and Archives, TX-56
University of Texas at El Paso. C.L. Sonnichsen Special Collections Department, TX-23
St. James Church, TX-57
U.S. National Archives at Fort Worth, TX-24
Our Lady of Perpetual Help Church, TX-58
Catholic Church. Archdiocese of Galveston-Houston. Archives and Records, TX-25
La Purisima Church, TX-59
Catholic Church. Diocese of Laredo. Chancery, TX-26
Livingston [La Porte]
Baylor University. Carroll Library. The Texas Collection, TX-60
San Jacinto Museum of History. Albert and Ethel Herzstein Library, TX-27


text:
Q: PhoneGap, Why an ajax post request doesnt works I'm using phonegap with jquery.
This works
$.ajax({url: 'samplefile.html'});

but this doesnt work
$.ajax({url: 'samplefile.html', type: 'POST'})

It returns a 404 error code. Why?

A: do like this:
$.ajax({
     type: "POST",
    url: "some.php",
     data: { name: "John", location: "Boston" }
    })

I think it's trying to retrieve html file thats why you are getting an error.
go through:  here

redpajama_set_name: RedPajamaStackExchange


text:
22 Brygada Artylerii (II RP)
 22 Brygada Artylerii Przeciwpancernej

redpajama_set_name: RedPajamaWikipedia


Fanfic by MMan in which Daria dreams that her younger sister Quinn turns into her younger brother, Quint. The story is often labeled an alternate universe tale because of its realism.
This page was last edited on 13 July 2013, at 03:25.


CenterPoint Energy Inc. is a natural gas and electricity utility in the United States of America. The company operates throughout Texas, Minnesota, Arkansas, Oklahoma, Louisiana and Mississippi. It was created in 1882, but it has been through a few different name changes.
It is now headquartered in Houston, Texas. It has a few divisions responsible for specific operations, but also more than 8,800 workers. In 2010, financial revenues got to $8.7 billion.
The net profits were slightly lower though – around $442 million. Some of the most important customers of CenterPoint Energy Inc. include Direct Energy, Champion Energy and First Texas Energy Corporation.


text:
Meg Ryan's "Serious Moonlight" Trailer
Posted: October 16th, 2009 by WorstPreviews.com Staff
The trailer for the upcoming comedy "Serious Moonlight," starring Meg Ryan and Timothy Hutton, has appeared online. Kristen Bell and Justin Long appear in the movie as well, and are clearly the highlight of it. Check out the trailer below.
Plot: After she arrives at her country home for a romantic weekend getaway, things don't go exactly as planned for high-powered Manhattan lawyer Louise (Ryan). First, her husband of 13 years, Ian (Hutton), tells her that he's leaving her for a younger woman (Bell). Then, one thing leads to another, and Ian finds himself held captive by Louise who explains that she won't release him until he professes his love for her.
The new movie is directed by actress Cheryl Hines ("Curb Your Enthusiasm"), who is making her feature film directorial debut. It is scheduled to hit theaters on December 4th, in limited release.
Click here to read more about "Serious Moonlight."
Source: Yahoo! Movies
The Dini writes:
on October 16th, 2009 at 8:59:05 AM
Meg Ryan is STILL HOT! Also, why is he Mac guy in EVERY MOVIE??!!
on October 16th, 2009 at 10:56:39 AM
Its a compiration mac wants to take the world and meg ryan needs a boob Job!
Clarence Worley writes:
on October 16th, 2009 at 12:44:41 PM
Serious Moonlight? I thought this was French Kiss 2...
Looks like she finally got her face evened out. She's been out of the spotlight the past few years because of some botched plastic surgery.
Rich k writes:
on October 16th, 2009 at 3:28:16 PM
I didnt even watch the trailer because the plot above was to f*cking stupid.
Arlo11 writes:
If I was a woman i'd be totally offended by this movie! :P oh and Justin Long is a funny mofo! THATS why hes in every movie!
the only reason i wanted to watch the trailer was cause of Justin Long... and apparently its been "removed" lol how convenient


text:
Q: How do you uninstall kerberos from Bash? or Change server name? I setup the wrong server name and admin name. I sthere a way to change it? instead of uninstalling it? Thanks

A: If you're running Debian or Ubuntu, then dpkg-reconfigure krb5-config will pop up the same "initial setup" dialogs as you get when installing the package. (Alternatively, apt purge would uninstall the package and remove all of its configuration.)
But in pretty much all systems, you can just edit the file /etc/krb5.conf and set your realm name and KDC & kadmin server addresses in there. For example:
[libdefaults]
    default_realm = EXAMPLE.COM

[realms]
    EXAMPLE.COM = {
        kdc = kerberos.example.com
    }

This is the same for both MIT Krb5 and Heimdal Kerberos.
(Though on a properly set-up realm, the KDCs can be discovered through DNS and don't need to be manually configured in each client's krb5.conf.)

If you were trying to set up a new MIT Kerberos KDC server, its database is defined in /var/lib/krb5kdc/kdc.conf (this might sometimes be placed in /etc/krb5kdc instead). Edit the realm name, delete the files storing the old database, then create a brand new realm using kdb5_util create -s.
(Heimdal uses different paths for the KDC configuration, and probably kadmin --local to initialize a new realm.)


text:
-   () est un album de remix du chanteur mexicain Luis Miguel. L'album a fait ses débuts à la  place pour ensuite se hisser à la place du  mexicain. Il a été certifié or et platine pour la vente de plus de . Du côté des États-Unis, l'album a été classé pendant une semaine à la  place du Billboard 200 et a atteint la  place du hitparade .

Contenu
Le , Luis Miguel revient avec un nouvel album, , une œuvre qui surprendra ses fans puisqu'il s'agit de dix de ses tubes pop rythmés, remixés.

Sur l'album, on trouve la chanson la plus connue en anglais que Luis Miguel réinterprète, , une chanson originale des Jacksons Five (Michael Jackson et ses frères), dont le nom a été changé en . Au-delà des paroles, le montage est identique à l'original enregistré en 1978.

Dave Shim, rédacteur d'AllMusic, donne une appréciation positive de l'album, affirmant que   et que .

L'album a débuté à la  place du classement Billboard 200, aux États-Unis. Le titre remixé  a été extrait comme premier single de cette collection. La chanson a rapidement atteint la première place des charts radio nationaux, dès le premier jour de sa sortie au Mexique ;  est également devenue l'album le plus vendu grâce aux téléchargements numériques après sa sortie.

Liste des pistes
Adapté de Discogs et d'AllMusic.

Références

Annexes

Articles connexes

 Luis Miguel

Liens externes

 Site officiel de Luis Miguel

Album musical sorti en 2010
Album de Luis Miguel
Album de pop
Album de musique latino-américaine


This book compiles textile terms from the Philippine colonial period, drawing on both primary and secondary sources to trace histories and use. The abaca fabric called medriñaque, for example, appears to have been used for men's shirts in 16th-century Peru and was recorded in 17th-century English records as "medrinacles," a coarse canvas used for sails or for stiffening collars and doublets.
Castro contextualizes the lexicon with a historical survey of the colonial trade and economy, which, in turn, shaped Philippine textile culture. With photographs of sample fabrics, the book serves as an introduction to those interested in textile history.


text:
Dec. 26, 2012 / 2:31 PM
Firm to open phone lines for job seekers
CHICAGO, Dec. 26 (UPI) -- Outplacement firm Challenger, Gray & Christmas in Chicago said it would be open for free call-in counseling for job seekers Thursday and Friday.
The firm said it would host its 27th annual call-in service for "anyone who is looking for a job, whether they are unemployed, wanting to switch jobs or re-entering the job market after a long absence."
Challenger, Gray & Christmas specializes in finding jobs for people who are sent to the firm by former employers. But for two days of the year for the last quarter of a century, the firm has opened its phone lines to one and all as a free service.
"Call-in is the only time that anyone can get help from the firm's professional job search coaches," the firm said in a statement.
The number for free job seeking advice is 312-422-5010. The phone-in job fair runs Dec. 27-28 from 9 a.m. to 5 p.m. CST.
Job candidates are not always forgiving Study: Internet reduces unemployment time Job-seekers vulnerable to fraud
Gray & Christmas
Defense News // 2 months ago
Austin visits Ukraine, voices U.S. support against Russian aggression
Oct. 19 (UPI) -- U.S. Defense Secretary Lloyd Austin said Ukraine continues to have U.S. support against Russian aggression and is free to decide its own foreign policy during a visit to Kyiv Tuesday.
Citing Russian occupation, U.S. signs new defense pact with Georgia
Oct. 18 (UPI) -- U.S. Defense Secretary Lloyd Austin signed a new security agreement with Georgia Monday, seeking to buttress relations with the Black Sea nation as it continues to face Russian troops in two of its regions.
Pentagon orders all civilian employees to be fully vaccinated by Nov. 22
Oct. 4 (UPI) -- The Pentagon on Monday ordered all Department of Defense civilian employees to be fully vaccinated against COVID-19 by no later than Nov. 22.
Army investigating suspected suicides of three Fort Drum soldiers
Sept. 29 (UPI) -- The U.S. Army is investigating the suspected suicides of three soldiers based at New York's Fort Drum.
Four Marines remain hospitalized month after suicide bombing at Afghan airport
Sept. 24 (UPI) -- Four U.S. Marines remain hospitalized after an attack near Hamid Karzai International Airport in Kabul, Afghanistan, last month as service members were evacuating foreign nationals and refugees.
South Korean firm to supply rapid COVID-19 test kits to U.S. military
SEOUL, Sept. 24 (UPI) -- South Korean biopharmaceutical firm Celltrion has agreed to supply its DiaTrust rapid COVID-19 test kit to the U.S. military.
Army tests MK-22 Precision Sniper Rifle at Fort Bragg ahead of fielding
Sept. 23 (UPI) -- The Army has successfully tested the MK-22 Precision Sniper Rifle to clear it for fielding.
U.S. Navy, Bahrain partner to ramp up unmanned systems use
Sept. 23 (UPI) -- Bahraini leaders committed Thursday to partnering with a new U.S. Navy task force to ramp up new unmanned system efforts.
U.S. F-16 fighter jets intercept Cessna during U.N. General Assembly
Sept. 22 (UPI) -- U.S. fighter jets escorted a small plane flown by a U.S. Army soldier out of restricted air space over New York City during the United Nations General Assembly.
U.S. Space Force to take over SATCOM operations from Army, Navy
Sept. 22 (UPI) -- The U.S. Space Force will take over satellite communications billets, funding and mission responsibility from the U.S. Army and Navy, the Department of Defense announced Wednesday.


Neeraj Pandey's 15-minute short film, Ouch, is a dark comedy which neatly turns the tables on the man-woman equation through a tongue-in-cheek take on an extramarital affair.
The film kicks off with Manoj Bajpayee aka Vinay strutting through the lobby of a five-star hotel, a middle-aged Casanova in tight jeans and dark glares, on his way to his usual rendezvous. His air of assurance and anticipation however dissipates like a balloon whose air has leaked out, once he is in his suite and the cell phone announces the intrusive presence of his mother with the ringtone, "Mummyji calling." The way he springs to his feet and greets her with a "pranam", grimacing at the white lie that "he's in a meeting", is the first sign that he's not the omni-potent male he pretends to be.
This realisation is further accentuated when his lover, Priya (Pooja Chopra), enters, walking straight into his arms, the proverbial damsel in distress. She's sweetly submissive, her sentences peppered with "I love you", her gaze sympathetic as she surveys the bandaged cut on Vinay's forehead, the result of his wife's frying pan fury after being told that he wants a divorce.
Priya's equally distraught but not because she's confessed to the office romance which blossomed through several bouts of 'relaxation' over the last three years. In fact, as she confesses, she couldn't bring herself to make a clean breast of things to her husband, Rakesh, as they'd decided. And in fact, after an attack of delayed conscience, has decided to bring an end to this "f***ing impossible dream".
That's the point in the narrative when Priya suddenly comes into her own, her "I love yous" taking on a different colour and interpretation, the four-letter word her colleague-paramour has been using so liberally, entering her vocabulary too. Vinay looks dazed as his life spirals out of control, three women — his wife, lover and mother — suddenly taking control. The ek-pe-ek free gift of a suitcase, which incidentally is a becoming shade of red, at the end is the chery on the cake.
As usual, the versatile and supremely talented Manoj is at his assured best, showcasing a range of emotions with effortless ease. Former Miss India and Commando girl Pooja Chopra is a perfect foil. Neeraj's sharp wit and assured direction makes a case for gender empowerment without the usual feminist chest-beating making this short an enlightening watch.
PS: The squeaking couch, a wry reference to B-wood's infamous casting couch, is a perfect ouch!


Hi, everyone! Very briefly, I wanted to let you know that I have made a slight change to the title of the digital boxed set edition of the first three books of The Inventor's Son. It will now be listed in the eBook stores as The Inventor's Son Collection (Books 0-2) because that is a much more accurate description of the contents. "Book 0" is commonly used for numbering a prequel book, but I've never been able to use it in the stores since they would not allow me to use anything but whole numbers. Since this set contains the prequel novella, Book 1 and Book 2, I wanted to make sure that the title of the set reflected that, now that I have gone and made sure the numbering of the volumes in all stores all correctly corresponds to the correct books.
The Inventor's Son: The Beginning, book 0, which has added material and is exclusive to the set (or to the mailing list).
The Inventor's Son, book 1 of the series, a full-length novel.
The Scientist's Son, book 2 of the series, a full-length novel as well.
The new retail price will be $4.99 in all stores, and in the coming days, there will be a pre-order set up for The Traitor's Son! Watch this space.


For students entering Point Park in the fall of 2019, the number of majors offered by the School of Communication will be fewer by four.
In a consolidation passed Monday by the faculty assembly, the existing broadcast reporting, environmental journalism, journalism, mass communication and photojournalism majors will be phased out in favor of a new, restructured bachelor of arts in journalism.
According to Christopher Rolinson, associate professor of photojournalism, the change comes on the heels of a decades-long consideration of integrating multi-platform reporting into the journalism curriculum.
The restructured program, consisting of 125 credits, combines the specialized elements of all five of the existing majors with the general requirements of the School of Communication.
The journalism faculty, chaired by Tim Hudson, passed their initial proposal on to the School of Communication Jan. 17. The journalism faculty consists of Hudson, Rolinson, Aimee Dorsten, Helen Fallon, Steve Hallock and Bill Moushey. Only Fallon voted against the measure.
The initial review of the broadcast reporting degree – which recommended the switch – consulted with a dean from a Florida university and four Point Park adjuncts: Maria Kakay, Gina Catanzarite, Nick Matoney and Susan Koeppen. Every five years, each degree undergoes an evaluation by faculty groups to ensure quality. While the reporting degree consulted with those professionals, Fallon said no working professionals were consulted on the restructuring.
In its proposal to the faculty assembly, the journalism department explains the rationale behind the restructuring as a shift in response to a changing industry and a paradigm shift in academia away from medium-specific majors.
The adoption signals the journalism faculty's intent to compete with programs offered at larger universities. In its research, the journalism faculty identified Penn State, Indiana University of Pennsylvania, Slippery Rock and California University of Pennsylvania as regional competition.
For his part, Rolinson pointed to the faculty-to-major ratio as part of the reasoning for the switch.
There is room for choices within the major requirements, such as between on-camera and radio announcing courses and a choice between specialized advanced reporting courses. Any chosen and prescribed specialization would be limited to 12 credits, or four classes, as major electives. While a student could create their own specialization, there are no formalized concentrations in the proposal.
Rolinson said there was a consideration for concentrations within the journalism degree, as the School of Communication already allows for a graphic design concentration in its multimedia degree.
Upon enrollment, each student is assigned a student success coordinator tasked with scheduling and a faculty advisor, whose role is to assist students in building a path toward employment.
According to Fallon, students often fail to forge that connection with their faculty advisor.
While the five journalism-based majors are being consolidated into a singular major, all classes currently available to the five majors will still be available. The realignment is not part of a budget cutback, and all six faculty members will remain part of the unified journalism program.
School of Communication chair Thom Baggerman sent an email to all current school of communication students clarifying that no current students will be impacted by this change.
The university is required to provide students the majors available upon each student's admission and enrollment in the university, meaning current students and next year's freshmen will not be affected. However, students may change to the new B.A. in journalism starting in the fall of 2019.
These changes do not affect the other majors in the School of Communication, including the bachelor of fine arts in photography, the broadcast production and media management and multimedia degrees.


text:
Marikina River Park
300 Shoe Ave, Santo Niño, Marikina, Philippines
14°37′47″N 121°5′3″E / 14.62972°N 121.08417°E / 14.62972; 121.08417Coordinates: 14°37′47″N 121°5′3″E / 14.62972°N 121.08417°E / 14.62972; 121.08417
220 hectares (540 acres)
Marikina River Park Authority
Santolan
https://www.marikina.gov.ph/
The Marikina River Park is a system of parks, trails, open spaces and recreation facilities along an 11-kilometer (6.8 mi) stretch of the Marikina River in Marikina, eastern Metro Manila, Philippines. The urban riverside park covers an area of 220 hectares (540 acres) running through the villages of Santo Niño, Santa Elena, Jesus de la Peña, San Roque, Calumpang, Barangka and Industrial Valley Complex in the western part of the city. The park, under development since 1993, is the city's biggest recreational and sports area containing the Riverbanks Center, a mall and office complex, and the biggest roller skating rink in the Philippines.[1] It is a popular area for camping, jogging, biking and other outdoor activities not just for Marikina residents but also for those from nearby cities in Metro Manila.
The park beside the Riverbanks Center
The park was developed as part of a rehabilitation program called Save the Marikina River Program by the Marikina city government. Launched in 1993, the program aims to reduce flooding and improve the Marikina riverbanks for better water discharge after a 1992 flood left 25% of the city under floodwater.[1] The program involved building access roads to the banks to allow for the cleaning and clearing operations, clearing the banks of informal settlers and other encroachments, demanding the local industries along the riverbanks to set up water treatment facilities in order improve the river's water quality, planting various flora and building cultural, sports and other historical structures to draw residents back to the river.[1]
Events and facilities
The Marikina River Park today is a focal point for community and cultural activities. It hosts the annual Mama Mary Fluvial Parade and Festival and the Marikina Christmas Festival.[1] It has an amphitheater, floating stages for cultural events, a Roman Garden, Chinese Pagoda, a gazebo, riverboats and picnic grounds. Sports facilities include a baseball field, a basketball court and a skating rink. A youth camp and a senior citizens lifestyle center can also be found in the park.[1]
Panorama of Marikina River Park
^ a b c d e "Flood Disaster Mitigation and River Rehabilitation by Marikina City, Philippines" (PDF). Asian Disaster Preparedness Center. Retrieved October 22, 2015.
Parks and plazas in Metro Manila
La Mesa Watershed Reservation
Las Piñas–Parañaque Critical Habitat and Ecotourism Area
Ninoy Aquino Parks and Wildlife Center
Quezon Memorial Circle
Rizal Park
Urban parks
APEC Sculpture Garden
Ayala Triangle Gardens
Baywalk
Bernardo Park
Bonifacio Shrine
Glorieta Park
Legazpi Active Park
Makati Park and Garden
Makati Poblacion Park
Malabon People's Park
Mehan Garden
Muntinlupa Sunken Garden
Navotas Centennial Park
Paco Park
Pinaglabanan Shrine
Salcedo Park
Sucat People's Park
Tandang Sora National Shrine
Valenzuela People's Park
Washington SyCip Park
Arroceros Forest Park
Balara Filters Park
Isla Pulo
La Mesa Ecopark
Malabon Zoo
Manila Zoo
Pasig Rainforest Park
University of the Philippines Arboretum
Squares and plazas
Liwasang Bonifacio
Plaza Cervantes
Plaza Dilao
Plaza de España
Plaza Lacson
Plaza Mexico
Plaza Miranda
Plaza Moraga
Plaza Moriones (Intramuros)
Plaza Moriones (Tondo)
Plaza Olivia Salamanca
Plaza Rajah Sulayman
Plaza de Roma
Plaza Rueda
Plaza San Lorenzo Ruiz
Book Stop Intramuros
Parks in Metro Manila
Buildings and structures in Marikina
Redeveloped ports and waterfronts in the Philippines


A lot of people make the decision to move themselves. Sometimes it's from a cost standpoint, or sometimes it's because they may not trust movers with their items. Most of the time, it's the cost though. We find that many people think they can't afford to have a mover come help, but let's break down the cost of your DIY move so you can see how much it really costs you.
Depending on the distance of your move and how much you have to move (for example, how rooted are you? Do you have tons of heavy expensive furniture you have to move), DIY can seem like the least expensive option. What should you budget for if you're planning to do your move on your own?
A truck rental can be anywhere from $19.95 an hour to more. You're going to want to find out the truck that can actually hold all your items (so you're not making multiple trips) and find out all of the associated costs with having your own moving truck. That may include a per-mile cost, usually around $1 per mile. Fuel is also not included in your truck rental price, so figure into the cost of refilling your truck. You may also opt into insurance for the truck as well for added peace of mind.
Don't forget, buying boxes can start to add up. Purchasing nice, new boxes can cost a lot if you're using a lot of them, especially if you opt into specialty boxes like wardrobe boxes. You may also be purchasing blankets or padding material to help insulate your valuables.
If you're making a long distance move, you may have to stay at a hotel overnight. Consider the cost of the hotel room you'll be sleeping in, and consider that the hotel room must allow you to park your moving truck too.
Chances are, you won't want to cook on packing day or moving day, so you might want to consider the cost of takeout or delivery to your overall expenditure during your move.
Remember that moving your things will cost you a lot of, you guessed it, time. Scheduling time to rent the truck, to actually pack your items at home, and then to move those boxes and items into the truck, along with the actual time it takes to drive your items and then unload them? That's a lot of time, especially if you have to take time out of work.
Want a little help? Contact us today to get started!


Set what is at rest in motion and there is no rest.
Just stop. That's all. Just stop.
All of the tendencies and habits that are restlessly pushing and pulling will just go on and on if you don't stop.
Then just don't stir yourself up. Just rest in this moment.
Just sit up straight and be without difficulty.
How often we rush through our lives. Where are we going? Do we realize the price we pay for this rushing about? Try stopping. See what happens.


Side Transfers from Antalya Airport £30 / Antalya Airport to Side Taxi Transfer upto 3 people £30 per way. Book your Side Transfers from Antalya Airport at low prices.
Antalya Airport to Kemer Taxi Transfer £30 per way upto 3 people. / Cheap Antalya Kemer Taxi Transfers / Antalya Airport Bus Transfers Available.
Turkey Airport Transfers are one of the most popular product that travel companies offer. There are 30 + Airports located in Turkey. But the most of the airport transfer companies operate in the Popular Tourist Airports mainly located in South West of Turkey.
Turkey Airport Transfers prices vary. Depending on the area you travel, Depending on the service type you book and ofcourse the distance you are travelling. In some cases, the near area transfer prices are higher than the remote areas because the resort you are travelling to / from may not receive demand enough to be able to run regular transfer services or it may be in totally reverse area.
The idea of offering Low-Cost Airport Transfers in Turkey is to provide economic prices as well as quality Turkey Airport Transfers. HolidayHoppa is here since 2007 providing Turkey Airport Transfers at low costs.
How Can a transfer company offer Low-Cost Turkey Airport Transfers ?
The answer to this question is simple but meaningful. If a transfer company can use the same transfer vehicle back to back then they can offer low prices. Basicly it means, the transfer taxi which takes you to your hotel, brings other people back to the Airport. Sometimes this isnt possible ofcourse, then the numbers of Turkey Airport Transfers operated by that company gets involved in order to offer low cost Turkey Airport Transfers.
Booking your Turkey Airport Transfers with HolidayHoppa is made easier. There are 3 main steps you need to follow. We will email your Turkey Airport Transfer voucher within the minutes after your booking if you book in the office hours. If you book your Turkey Airport Transfers outside the office hours, We will email your confirmation voucher in the next day.
What do i need to do after i arrive into my destination Turkey Airport ?
The information you need is printed on your confirmation voucher. You only need to read through your voucher and follow the steps. There are agent desks we use in some Turkey Airports. And in some of them we greet you with your name sign outside the arrivals exit gate. Therefore we advise you to read your voucher carefully. And take a print or digital copy with you for your Turkey Airport Transfers.
Book it in 3 easy Steps and see us on your arrival.


text:
Titan Industries - Investment & Expansion Plans
Titan Industries has plans to open 300 retail stores for its 'Titan Eye' in the next three years across the country, a senior company official said today.
The company would add another 20 exclusive optical showrooms in the current fiscal, Titan Industries Integrated Retail Services Vice President V Govindaraj told reporters here after inaugurating the first 'Titan Eye' store in the city, the fifth in Tamil Nadu and 40th in India.
As the current Indian prescription eyewear market was "sizeable" with nearly 30 per cent of the population requiring vision correction, the company was eyeing a substantial market, with Rs 50 crore in 2009-10 and Rs 300 crore to Rs 500 crore in the third year, Govindaraj said.
Titan industries, a joint venture between Tata group and Tamil Nadu Industrial Development Corporation (TIDCO), has reported a turnover of Rs 3,041 crore for the year ended 2007-08, he said, adding that Titan Eye has recently entered into a technical pact with Sankara Nethralaya for training of retail and clinic staff.
The company was also running Titan Watches and Tanishq brand of jewellery, besides a precision engineering unit for supplying spares to aerospace and automobile industries, he said.
With a market size of Rs 1,500 crore to Rs 2,000 crore, the company expects to capture a substantial eyewear market share in the near future, Govindaraj said.
As per Sep 08 shareholding of the Titan Industries, Rakesh Jhunjhunwala is holding 2,581,062 shares of Titan Industries in the name of Jhunjhunwala Rakesh Radheshyam.
Labels: Best Stocks To Buy, Growth Stock


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Jeff Bezos Alleges American Media's Deputy General Counsel Attempted To Extort Him
Amazon CEO Jeff Bezos said American Media Inc. threatened to release personal photos of him if he did not comply with a set of demands. He published an email from the New York-based company's deputy general counsel Thursday.
By Caroline Spiezio | February 07, 2019 at 07:38 PM | The original version of this story was published on Corporate Counsel
Jeff Bezos, founder and chief executive officer of Amazon.com Inc., listens during an Economic Club of Washington discussion in Washington, D.C., on Sept. 13, 2018.Photographer: Andrew Harrer/Bloomberg
Amazon founder and chief executive officer Jeff Bezos has alleged American Media Inc., which owns the National Enquirer, attempted to extort him by threatening to release personal photos unless he dropped an investigation of the company.
Caroline Spiezio
Caroline covers the intersection of tech and law for Corporate Counsel. She's based in San Francisco. Find her on Twitter @CarolineSpiezio.
Tech to the Rescue: Legal Departments Turn to Modern Tools
Victoria Hudgins | November 01, 2019
Outside and in-house counsel see legal tech as a dependable ally in delivering proven efficiency during the throes of a recession.
More Real Estate Deals Could Be Swept Up Under Proposed New Regulations for CFIUS Reviews
MP McQueen | September 25, 2019
The proposals for expanded review by the Committee on Foreign Investment in the U.S., the interagency panel at the Department of the Treasury, could catch some real estate investors and funds off-guard, international trade and national security lawyers said.
Data Security Firm Druva Hires Former Apple, Broadcom Lawyer as Its First General Counsel
Phillip Bantz | October 29, 2019
Jung-Kyu McCann McCann joined Sunnyvale, California-based Druva in September as the firm's first general counsel, according to a company spokesman. She commented on her goals for the company.


After your magnolia tree matures and starts getting very wide and tall, you may wonder whether or not to prune it and, if so, how and when to do so. There are some general rules to follow here that will give you guidance. But there are also exceptions to the rules.
Do so just after flowering is done, not only so you can at least enjoy the current year's blooms, but also because magnolia trees bloom on old wood (which means that pruning too late in the year puts you at risk for losing the buds for next year's flowers).
Trim off only a small amount, with an eye to shaping the plant to your liking.
Make your cuts with pruners or loppers that have first been thoroughly disinfected (for example, with ethanol or isopropyl alcohol) and sharpened.
Cut off dead branches first. Next, inspect the plant for areas where two branches are rubbing against each other; decide which one will lend itself better to the overall shape you are aiming for, then remove the other one. Finally, prune off any limbs that you suspect are infected by a disease (you want to make sure that the disease does not spread). Disinfect your cutting tool once again after this part of the job is done and before continuing to prune.
Make clean cuts with your pruners. Prune all of the way through the wood. Do not leave a branch hanging "by a thread" and then attempt to yank it off with your hand. In the process, you could end up ripping some bark off the trunk.
During a severe wind storm (or snowstorm or ice storm), a huge pine bough falls on your Jane magnolia tree (Magnolia Jane).
An accidental "pruning" results from the damage and the tree's leader is broken, altering its shape and growth habit forever.
Your Jane magnolia takes on a shrub form thereafter.
After such damage from a winter storm, you may wish to change your approach to how you prune the magnolia. Accepting the fact that it will now be growing as a shrub (not a tree), maybe you will now want to prune it so as to give it the most pleasing shape that you can within the limits of its new plant form. The tree form may have been the ideal, but it is advisable to adapt your expectations and your pruning regimen to the new reality.
By pruning the damaged plant judiciously, at least you will be able to maintain it as a compact shrub that fits neatly into a designated nook or cranny in your landscape. Depending on your personal tastes, while the "wild look" may suit a magnolia just fine in tree form, a more manicured look may suit the plant better in shrub form.
By taking advantage of the fact that the plant is just starting out, pruning can ensure that the mature specimen will have a shape suited to your tastes. For example, remove low branches if you prefer a tree form. That way, energy will go into creating stronger upper branches.
This early pruning can also promote better flowering.


Q1. Valery Bykovsky passed away at the age of 84 years. He was a_____________.
Sol. Russian cosmonaut Valery Bykovsky, who made the first of his three flights to space in 1963, passed away at the age of 84 years.
Q2. Who among the following will be awarded the prestigious Bodley Medal, the highest honor granted by the University of Oxford's world-famous Bodleian Libraries?
Sol. Nobel Prize-winning economist Amartya Sen will be awarded the prestigious Bodley Medal, the highest honor granted by the University of Oxford's world-famous Bodleian Libraries.
Q3. Who among the following has/have been honored with the Turing Award?
निम्नलिखित में से किसे ट्यूरिंग अवार्ड से सम्मानित किया गया है?
Sol. Three pioneers in AI, University of Toronto faculty member and Google Brain researcher Geoffrey Hinton, Facebook chief AI scientist and NYU professor Yann LeCun, and Element AI founder and University of Montreal professor Yoshua Bengio has been honored with the Turing Award.
Q4. Name the first Indian port operator that has recorded cargo movement of more than 200 million metric tonnes (MMT) at nine ports along the east and west coasts.
Sol. Adani Ports and Special Economic Zone (APSEZ) announced that it has recorded cargo movement of more than 200 million metric tonnes (MMT) at nine ports along the east and west coasts, becoming the first Indian port operator to achieve the milestone.
Q5. Who among the following is the author of the book "Indian Fiscal Federalism"?
निम्नलिखित में से कौन "इंडियन फिस्कल फेड़ेरेलिस्म" पुस्तक के लेखक है?
Sol. The book "Indian Fiscal Federalism" has been written by Dr. Y. V. Reddy, former Governor of RBI and Chairman of the 14th Finance Commission of India along with Dr. G. R. Reddy, Advisor (Finance) to the Government of Telangana.
Q6. Which of the following organisation has unanimously passed the first-ever resolution ordering members to enforce laws against terror financing?
निम्नलिखित में से किस संगठन ने सर्वसम्मति पहली बार सदस्यों को आतंकी वित्तपोषण के विरुद्ध कानून लागू करने का आदेश दिया है?
Sol. The UN Security Council has unanimously passed the first-ever resolution ordering members to enforce laws against terror financing.
Q7. State-owned lender Punjab National Bank (PNB) had sold stake in its housing finance arm, PNB Housing Finance (PNBHF) to General Atlantic Group and Varde Partners for___________.
Sol. State-owned lender Punjab National Bank (PNB) had sold stake in its housing finance arm, PNB Housing Finance (PNBHF) to General Atlantic Group and VardePartners for Rs 1,851 crore at Rs 850 per share.
Q8. Name the Himachal Pradesh village that has become the highest polling station in the world.
Sol. Tashigang, a Himachal Pradesh village at an altitude of 15,256 feet has now become the highest polling station in the world. Situated about 29 km from the India-China border, the polling station covers two villages, Tashigang and Gate.
Q9. Who among the following will now serve as the Ethics Officer of the Indian cricket board on an ad-hoc basis?
निम्नलिखित में से कौन अब एक तदर्थ आधार पर भारतीय क्रिकेट बोर्ड के एथिक्स अधिकारी के रूप में कार्यभार संभालेंगे?
Sol. Supreme Court-appointed BCCI Ombudsman Justice D K Jain (retd) will now serve as the Ethics Officer of the Indian cricket board on an ad-hoc basis.
Q10. The Finance Ministry has decided to infuse _____ into state-owned Bank of Baroda (BoB) ahead of the merger of two other public sector lenders Dena Bank and Vijaya Bank with BoB.
Sol. The Finance Ministry has decided to infuse Rs 5,042 crore into state-owned Bank of Baroda (BoB) ahead of the merger of two other public sector lenders Dena Bank and Vijaya Bank with BoB.