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MOLECULAR AND CELLULAR PATHOGENESIS
Glycosylation of Homologous Immunodominant Proteins of Ehrlichia chaffeensis and Ehrlichia canis
Jere W. McBride, Xue-jie Yu, David H. Walker
Jere W. McBride
Department of Pathology and WHO Collaborating Center for Tropical Diseases, University of Texas Medical Branch, Galveston, Texas 77555-0609
Xue-jie Yu
David H. Walker
DOI: 10.1128/IAI.68.1.13-18.2000
The glycoprotein genes of Ehrlichia chaffeensis (1,644 bp) and Ehrlichia canis (2,064 bp) encode proteins of 548 to 688 amino acids with predicted molecular masses of only 61 and 73 kDa but with electrophoretic mobilities of 120 kDa (P120) and 140 kDa (P140), respectively. The 120-kDa protein gene of E. chaffeensis contains four identical 240-bp tandem repeat units, and the 140-kDa protein gene of E. canis has 14 nearly identical, tandemly arranged 108-bp repeat units. Conserved serine-rich motifs identified in the repeat units of P120 and P140 were also found in the repeat units of the human granulocytotropic ehrlichiosis agent 130-kDa protein and of the fimbria-associated adhesin protein Fap1 ofStreptococcus parasanguis. Nearly the entire (99%)E. chaffeensis P120 gene (1,616 bp), the 14-repeat region (78%) of the E. canis P140 gene (1,620 bp), and a 2-repeat region from the E. chaffeensis P120 gene (520 bp) were expressed in Escherichia coli. The recombinant proteins exhibited molecular masses ranging from 1.6 to 2 times larger than those predicted by the amino acid sequences. Antibodies against the recombinant proteins reacted with E. chaffeensis P120 andE. canis P140, respectively. Carbohydrate was detected on the E. chaffeensis and E. canis recombinant proteins, including the two-repeat polypeptide region of E. chaffeensis P120. A carbohydrate compositional analysis identified glucose, galactose, and xylose on the recombinant proteins. The presence of only one site for N-linked (Asn-Xaa-Ser/Thr) glycosylation, a lack of effect of N-glycosidase F, the presence of 70 and 126 Ser/Thr glycosylation sites in the repeat regions of P120 and P140, respectively, and a high molar ratio of carbohydrate to protein suggest that the glycans may be O linked.
Ehrlichia chaffeensis andEhrlichia canis are obligate intracellular bacteria that exhibit tropism for monocytes and macrophages and are responsible for the diseases human monocytotropic ehrlichiosis and canine ehrlichiosis, respectively (10). Recently, the 120- and 140-kDa protein genes from E. chaffeensis and E. canis have been cloned, expressed, and characterized (18, 19). The 120-kDa protein (P120) of E. chaffeensis and the 140-kDa protein (P140) of E. canis are immunodominant and E. chaffeensis P120 appears to be surface exposed (9a). The proteins are homologous, and each has a region of serine-rich tandem repeats. The recombinant E. canis P140 and E. chaffeensis P120 exhibit molecular masses much larger than those predicted by the amino acid sequences, and antibodies produced against the recombinant proteins recognized native E. chaffeensisand E. canis proteins of similar sizes (18, 19). Two proteins (P100 and P130) from the human granulocytotropic erhlichiosis (HGE) agent have also been cloned, and the recombinant proteins exhibited higher-than-predicted molecular masses (12).
The existence of glycoproteins in eukaryotic cells has been known for years and was thought to be restricted to these cells. However, more recently, glycoproteins have been identified in eubacteria and archaebacteria (6, 7). The best characterized prokaryotic glycoproteins are surface layer (S-layer) proteins, but other membrane proteins unrelated to the S-layer have also been reported in eubacteria, such as species of Streptococcus,Mycobacterium, and Borrelia (4, 11,15). Characterization of eubacterial glycoproteins thus far has revealed diverse carbohydrate structures compared to eukaryotes, which have more conserved glycan structures. Many specific functions of glycoproteins in prokaryotes have been reported, including maintenance of cell shape, protein stability, protection against proteolysis, and adherence to target cells (7).
Glycans are usually attached to proteins by an asparagine residue (N glycosylation) or to a serine or threonine (O glycosylation). Both types of glycosylation have been reported for eubacterial S-layer and outer membrane proteins (7). N- and O-linked glycans typically consist of a conserved core region with variable branching sugars, but glycans without a conserved core region have also been reported (8).
The purpose of this study was to determine whether the recombinant and native proteins from E. canis and E. chaffeensisare glycosylated. The extent and character of the glycans on the ehrlichial proteins were examined by using carbohydrate compositional analysis, glycosidases, and lectins to better understand the glycan structure on recombinant and native ehrlichial glycoproteins.
Ehrlichiae and purification. E. canis (Oklahoma isolate) and E. chaffeensis (Arkansas strain) were provided by Jacqueline Dawson (Centers for Disease Control and Prevention, Atlanta, Ga.). Propagation of ehrlichiae was performed in DH82 cells with Dulbecco modified Eagle medium supplemented with 10% bovine calf serum and 2 mM l-glutamine at 37°C. The intracellular growth in DH82 cells was monitored by the presence of E. canis and E. chaffeensis morulae by using general cytologic staining methods. Cells were harvested when 100% of the cells were infected with ehrlichiae and were then pelleted in a centrifuge at 17,000 × g for 20 min. Cell pellets were disrupted with a Braun-Sonic 2000 sonicator twice at 40 W for 10 s on ice. The lysate was loaded onto discontinuous gradients of 42, 36, and 30% Renografin (E. chaffeensis) or onto a 32% continuous Percoll gradient (E. canis) as previously described (14). Ehrlichiae were collected and washed with sucrose-phosphate-glutamate buffer (218 mM sucrose, 3.8 mM KH2PO4, 7.2 mM K2HPO4, 4.9 mM glutamate, pH 7.0) and pelleted by centrifugation.
Gene cloning and expression and purification of recombinant proteins.The E. chaffeensis P120 gene (1,864 bp) (including a 260-bp noncoding region corresponding to the carboxy terminus), the E. canis P140 gene (1,620 bp), and a region consisting of two repeat units (520 bp) from the E. chaffeensis P120 gene were cloned into pGEX expression vectors as previously described (17-19). All constructs were transformed into Escherichia coli BL21, and the recombinant fusion proteins were expressed after IPTG (isopropyl-β-d-thiogalactopyranoside) induction for 4 h at 37°C. Bacteria were harvested by centrifugation at 5,000 × g for 20 min and resuspended in phosphate-buffered saline with protease inhibitors (Roche Molecular Biochemicals, Indianapolis, Ind.). The suspension was sonicated to lyse the bacteria, and the insoluble material was pelleted by centrifugation. The supernatant was removed, and the soluble recombinant proteins were purified by using glutathione-Sepharose (Amersham Pharmacia Biotech, Piscataway, N.J.).
Detection of native E. chaffeensis and E. canis glycoproteins.Purified E. chaffeensis andE. canis organisms grown in vitro were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the separated proteins were transferred to nitrocellulose. The membrane was incubated in blocking buffer (1% nonfat milk, 0.1 M Tris, pH 7.5) for 1 h. Blots were incubated with homologous and heterologous rabbit (E. chaffeensis P120) or mouse (E. canis P120) antisera against the respective recombinant proteins at 1:100 for 1 h. Bound antibody was detected with a secondary goat anti-rabbit or goat anti-mouse immunoglobulin G (α-chain) alkaline phosphatase-labeled conjugate (Kirkegaard & Perry Laboratories, Gaithersburg, Md.) and visualized with 5-bromo-4-chloro-3-indolyl phosphate–nitroblue tetrazolium (BCIP-NBT) substrate.
Detection of glycoproteins.Glycoprotein detection was performed with a glycoprotein detection kit according to the protocol of the manufacturer (Bio-Rad Laboratories, Hercules, Calif.). Briefly, recombinant proteins were incubated in solution with periodate, oxidizing adjacent hydroxyl groups in carbohydrates to generate free aldehyde groups. The oxidized aldehydes were subsequently labeled with biotin hydrazide. The labeled proteins were subjected to SDS-PAGE and transferred to nitrocellulose. The membrane-bound biotin-labeled glycoproteins were detected with alkaline phosphatase-labeled streptavidin and visualized with BCIP-NBT substrate.
Monosaccharide analysis.Analysis of the monosaccharide ratio and total carbohydrate from E. chaffeensis P120 andE. canis P140 recombinant proteins and the glutathioneS-transferase (GST) protein was performed by gas chromotography. Purified fusion proteins (100 μg) were cleaved with thrombin for 24 h, dialyzed against water, and dried. Samples were hydrolyzed with 1 M methanolic HCl for 16 h at 80°C. The released sugars were derivatized with Tri-Sil, and the samples were analyzed on a gas chromatograph with a Supelco column.myo-Inositol (20 μg) was added as a standard. The molar ratio of carbohydrate to protein was determined based on the quantity of recombinant protein submitted for analysis (100 μg), as estimated with stained SDS-polyacrylamide gels. Monosaccharide analysis was performed at the University of Georgia Complex Carbohydrate Research Center, Athens.
Lectin analysis.Lectin-binding analysis was performed with recombinant proteins and native E. canis and E. chaffeensis proteins by using a glycan differentiation kit (Roche Molecular Biochemicals) according to the manufacturer's protocol. The following digoxigenin-labeled lectins were used: Galanthus nivalis agglutinin recognizes terminal mannose linked α(1-3), α(1-6), or α(1-2); Sambucus nigra agglutinin recognizes sialic acid linked α(2-6) to galactose; Maackia amurensisagglutinin recognizes sialic acid linked α(2-3) to galactose; peanut agglutinin recognizes core disaccharide galactose β(1-3)N-acetylgalactosamine; and Datura stramoniumagglutinin recognizes Gal α-(1-4) GlcNAc. Each was incubated withE. canis, E. chaffeensis, and uninfected DH82 cells separated by SDS-PAGE and with purified recombinant proteins. Lectins were detected with an antidigoxigenin alkaline phosphatase-labeled antibody and visualized with BCIP-NBT substrate.
Enzymatic deglycosylation.Analysis of glycan linkages ofE. chaffeensis P120 and E. canis P140 was performed with a panel of glycosidase enzymes. Recombinant proteins were incubated with endoglycosidases (N-glycosidase F andO-glycosidase) and exoglycosidases (neuraminidase, β-galactosidase, and glucosaminidase) supplied in a enzymatic deglycosylation kit under denaturing conditions according to the protocol of the manufacturer (Bio-Rad Laboratories). The recombinant proteins were incubated independently with the enzymes to examine specific glycan linkages.
Protein analysis.Amino acid motifs were identified by using a Clustal alignment, and Emini surface probability plots were performed with protein sequence analysis software (Protean; DNAStar, Madison, Wis.).
Expression of recombinant proteins in E. coli. TheE. chaffeensis P120 gene (1,864 bp; partial), the E. canis P140 gene (1,620 bp; partial), and the E. chaffeensis repeat region (520 bp) were cloned into pGEX expression vectors (Fig. 1). The recombinant proteins expressed in E. coli BL21 cells exhibited molecular masses 1.6 to 2 times larger than those predicted by the amino acid sequence for each gene. The predicted molecular mass of the E. chaffeensis P120 recombinant protein was 67 kDa, that of the E. canis P140 protein was 57 kDa, and that of the E. chaffeensis two-repeat region was 20 kDa. However, the 100-kDa molecular mass of recombinant E. chaffeensis P120 observed on SDS-PAGE was approximately 1.6 times larger than the protein size predicted by the amino acid sequence, and the recombinant two-repeat region of E. chaffeensis P120 (60 kDa) was also 1.6 times larger than predicted by the amino acid sequence (Fig. 2). The 14 tandem repeats of E. canis P140 exhibited a molecular mass of 112 kDa, or approximately double the size predicted by the amino acid sequence (Fig. 2). All molecular masses were determined by subtracting the mass of 28-kDa GST fusion protein from the observed molecular mass of the recombinant fusion protein.
Agarose gel electrophoresis of E. chaffeensisP120 gene and E. canis P140 gene pGEX constructs digested to release inserts containing incomplete E. chaffeensis P120 gene (1,864 bp; 99%) (lane 2) and E. canis P120 gene (1,620 bp; 78%) (lane 3). Lane 1, molecular size markers (1-kb ladder).
(A) SDS-PAGE of recombinant ehrlichial GST fusion proteins expressed in E. coli BL21 cells. Lane 1, E. canis P140; lane 2, E. chaffeensis P120; lane 3,E. chaffeensis P120 two-repeat units; lane 4, GST protein (28 kDa). (B and C) Specificity of anti-recombinant E. canisP140 (B) and anti-recombinant E. chaffeensis P120 (C) against recombinant E. canis P140 (lanes 1) and E. chaffeensis P120 (lanes 2) proteins after removal of GST by thrombin protease. Numbers on the left of each panel are molecular masses in kilodaltons.
Identification of native glycoproteins.Native proteins with molecular masses of 120 and 140 kDa from E. chaffeensis andE. canis, respectively, reacted with antisera against the corresponding homologous recombinant glycoproteins (Fig.3). Antiserum against each respective recombinant protein did not cross-react with the heterologous native protein (not shown).
Immunoblot showing electrophoretically separated proteins of E. canis and E. chaffeensisorganisms, demonstrating the reaction of E. canis P140 (lane 1) and E. chaffeensis P120 (lane 2) proteins with antisera produced against the corresponding recombinant proteins. The recognized ehrlichial proteins are approximately twice as large as those predicted by the amino acid sequences encoded by the genes. Numbers on the left are molecular masses in kilodaltons.
Glycoprotein detection.Recombinant proteins representingE. chaffeensis P120, E. canis P140, and the two-repeat region of E. chaffeensis P120 were analyzed for carbohydrate content. Recombinant E. canis P140, E. chaffeensis P120, E. chaffeensis P120 repeat, and GST proteins were labeled with biotin to detect carbohydrate residues (Fig.4). The GST fusion protein (28 kDa) was not labeled (not shown).
Detection of carbohydrate on the expressed recombinant fusion proteins after biotin labeling of oxidized sugars. Lane 1,E. canis P140 (arrow); lane 2, E. chaffeensisP120 and proteolytic degradation products (arrow); lane 3, E. chaffeensis P120 two-repeat units (arrow). Numbers on the left are molecular masses in kilodaltons.
Analysis of protein glycosylation sites.Analysis of predicted glycosylation sites on the E. canis P140 recombinant protein revealed only 2 possible N-linked glycosylation sites (amino acids 7 and 32; Asn-Xaa-Ser/Thr) and 147 possible O-linked glycosylation sites (Ser or Thr), and analysis of predicted glycosylation sites on E. chaffeensis P120 revealed 91 O-linkage sites and two N-linkage sites. The E. chaffeensis P120 and E. canis P140 proteins have a high serine-and-threonine content that is particularly concentrated in the repeat regions of the proteins (Table1; Fig. 5).E. canis P140 has 110 serine residues, of which 98 are located in the repeat region, and E. chaffeensis P120 has 67 serine residues, with 52 located in the repeat region. Threonine, another potential O-linked glycosylation site, was present at many sites in the repeat regions of each protein. E. chaffeensisP120 has 18 threonine residues in the entire repeat region, andE. canis P140 has 28 threonine residues in the repeat region. A total of 16 amino acids (20%) of 80 were identified as potential O-linked glycosylation sites on each E. chaffeensis P120 repeat, while E. canis P140 had 9 of 36 (25%) in each repeat. Although the conserved N-linked glycosylation motif Asn-Xaa-Ser/Thr was present in two locations on each protein, only one of these sites was present in each of the expressed recombinant proteins. Homologous serine- and threonine-rich motifs were identified in the repeat regions of E. chaffeensis P120,E. canis P140, HGE agent P130, and Streptococcus parasangius Fap1 (Fig. 6).
Predicted and observed molecular masses and serine-and-threonine contents of ehrlichial glycoproteins and repeat regions
Schematic representation of predicted surface-exposed regions as determined by an Emini surface probability plot of E. chaffeensis P120, E. canis P140, and HGE agent P130 repeat regions. Potential O-linked glycosylation sites within each repeat unit are underlined. A value of >1 represents an increased probability of surface location.
Clustal alignment of shared homologous serine-rich motifs within the repeat units of S. parasangius Fap1,E. canis P140, E. chaffeensis P120, and HGE agent P130. Shared identical amino acids are boxed, and potential glycosylation sites in each motif are shown in boldface.
Glycosidase analysis.Incubation of E. canis andE. chaffeensis P120 recombinant proteins withN-glycosidase F (N-linked glycans), O-glycosidase (Galβ1→3GalNAc), neuraminidase (sialic acid residues), β-galactosidase (β-linked 1-4 terminal galactose), or glucosaminidase (β-linked N-acetylglucosamine) did not result in deglycosylation as determined by a reduction in the electrophoretic mobilities of the recombinant proteins as evaluated by SDS-PAGE.
Lectin binding.Recombinant and native E. chaffeensis and E. canis proteins did not react with the lectins specific for conserved glycan core structures that were tested.
Monosaccharide composition and carbohydrate/protein ratio.Monosaccharides attached to the recombinant glycoproteins were identified by using gas chromatography. The molar percentages of the monosaccharides and the estimated molar ratios of carbohydrate to protein represented on E. chaffeensis P120 and E. canis P140 are shown in Table 2. Carbohydrate was not detected on the GST protein.
Carbohydrate compositional analysis of recombinantE. chaffeensis P120 and E. canis P140 and estimated carbohydrate-to-protein molar ratio
The existence of prokaryotic glycoproteins has been demonstrated in previous studies which characterized S-layer glycoproteins and other membrane-associated glycoproteins in prokaryotes (6, 7). The larger-than-predicted molecular masses of recombinant and nativeE. canis P140 and E. chaffeensis P120 appear to be a result of an unusual posttranslational O-linked glycosylation. The two-repeat region that was expressed from the E. chaffeensisP120 gene did not contain any N-linked glycosylation sites and still exhibited 1.6 times the predicted molecular mass of the protein. There was one N-linkage site for glycosylation in the other larger P120 and P140 recombinant proteins, but glycans could not be removed byN-glycosidase F, an enzyme specific for N linkages. Together, this evidence supports the attachment of glycans to P120 and P140 via O linkages.
The recombinant proteins expressed in E. coli were found to have three carbohydrate residues, i.e., glucose, galactose, and xylose. The compositional analysis did not reveal any conserved core sugars such as N-acetylglucosamine,N-acetylgalactosamine, or mannose. The sugar composition observed in the recombinant proteins suggests an unusual glycan modification. Others have reported O-linked glucose moieties on proteins such as epidermal growth factor, tissue plasminogen activator, and coagulation factors VII, IX, and XII. Xylose-glucose O linkages to serine residues have been described previously and appear to be similar to the composition and linkage observed on the ehrlichial recombinant proteins (8). A compositional analysis of the sugars found on recombinant P120 and P140 indicated that glucose was the primary residue on the recombinant proteins. There were differences in the molar percentage of sugars on each recombinant protein, a manifestation of the microheterogeneity that exists between the two glycoproteins. These findings also suggest that glucose may be the primary attachment via O linkage to serine or threonine. A more detailed structural analysis would elucidate the exact nature of the sugar chains or monosaccharide linkages.
We attempted to further characterize the glycan structure by utilizing lectins and glycosidases. Lectins and glycosidases recognize very specific motifs, and the negative results that we obtained with both approaches confirmed that the glycosylation on the native and recombinant proteins does not contain common core structures often found in N- or O-linked eukaryotic glycans. Lectins that recognizeN-acetylglucosamine or N-acetylgalactosamine failed to react with the recombinant or native E. chaffeensis P120 and E. canis P140. Lectins and glycosidases are most commonly used to characterize N- or O-linked glycoproteins, which contain conserved core structures. Characterization with lectins becomes more difficult when glycans lack these core structures. Our findings and those of others suggest that some prokaryotic glycoproteins exhibit unconventional glycan structures.
Our finding that the recombinant proteins were glycosylated by E. coli was contrary to previous reports. It has generally been accepted and demonstrated that E. coli is unable to glycosylate various proteins that are glycoproteins in a native state (1). However, a glycosylated pilus of E. coli has been reported and provides evidence that the organism is capable of protein glycosylation to some extent (13). Our observations of larger-than-predicted sizes of recombinant proteins in E. coli were also similar to those when recombinant proteins from the HGE agent were expressed (12). This evidence supports the concept that specific glycosylation modifications are performed byE. coli. The recent sequencing of the entire E. coli genome has also led to the identification of glycotransferase enzymes that may be involved in protein glycosylation (2). The inability of E. coli to glycosylate many eukaryotic recombinant proteins points to possible differences in biosynthetic pathways that may exist between eukaryotes and prokaryotes, or perhaps only proteins transported to the plasma membrane are glycosylated. It is clear that E. coli can glycosylate some proteins and may preferentially glycosylate certain proteins of prokaryotic origin.
The repeat regions of the E. chaffeensis and E. canis proteins appear to be targeted for glycosylation. Other large proteins (P100 and P130) with repeat regions cloned from the HGE agent also demonstrate larger-than-predicted molecular masses (12) and seem likely to be glycosylated. Carbohydrate has also been detected on the Fap1 fimbria-associated adhesin protein ofS. parasanguis (15). We identified several similar serine-rich motifs in the repeat regions of HGE agent P130,E. chaffeensis P120, E. canis P140, and S. parasanguis Fap1. The conservation of these serine/threonine motifs among the four repeat regions suggests that they may be the primary targets for glycosylation. The number of serine and threonine residues in each protein correlated with the magnitude of the increased molecular sizes of P130 and P140 compared to P120 of E. canis and P100 of the HGE agent. E. canis P140 and HGE agent P130 exhibit a higher ratio of observed-to-predicted molecular mass than E. chaffeensis P120, and these proteins have 5 to 14% more serine and threonine residues, respectively. O-linked glycosylation most commonly occurs on serine and threonine residues, although tyrosine is occasionally glycosylated (1). The high serine content found in the repeat regions of these proteins is most likely where the glycans are attached. Expression of the two-repeat region from E. chaffeensis P120 resulted in a protein 1.6 times larger than that predicted by the amino acid sequence. Extrapolation from the mass of the two-repeat region to the entire repeat region completely accounted for the increased molecular mass of the recombinant E. chaffeensis P120. This calculation suggests that glycosylation of the recombinant protein occurs in the serine-rich repeat region of the protein. The recombinant E. canis P140 protein exhibited twice the molecular mass predicted by the amino acid sequence. This difference was also more exaggerated than the increased molecular mass of E. chaffeensis P120.E. canis P140 does have more potential glycosylation sites in the repeat region; in addition, the repeat region of E. canis P140 is much larger than the repeat region of E. chaffeensis P120. These differences may account for the larger increase in the molecular mass of E. canis P140. The recombinant E. chaffeensis P120 was slightly smaller than the native protein recognized by antiserum raised against the recombinant protein. This suggests that glycosylation of the native glycoproteins may be slightly different in the number of sugars or character of the sugar residues.
The similarities among P120, P140, HGE agent P130, and the fimbria-associated adhesin protein Fap1 of S. parasangiussuggest that these proteins may have similar locations and functions. The majority of glycosylated eubacterial proteins described thus far are S-layer proteins or other proteins associated with the outer membrane (7). Glycosylation of P120 and P140 further supports previous data that P120 is an E. chaffeensissurface protein. The function of E. chaffeensis P120 andE. canis P140 has not been determined, but E. chaffeensis P120 has been localized to the surface of E. chaffeensis and the extracellular fibrillar matrix in the morula by immunoelectron microscopy (9a). This observation is consistent with the case for other prokaryotic glycoproteins which are surface or membrane associated (6). In addition, it appears that in prokaryotes glycosylation occurs at the outer side of the plasma membrane (5) where surface proteins would be transported prior to their final destination. An S-layer morphology has been observed on the surface of Rickettsia prowazekii(9), and a protein believed to be an S-layer protein has been described (3). Most S-layer proteins described thus far contain a signal sequence and a high proportion of acidic amino acids; however, the S-layer protein identified in R. prowazekiidoes not contain a signal sequence, nor does E. chaffeensisP120 or E. canis P140, but these proteins contain a high proportion of acidic amino acids as often found in S-layer proteins. These similarities suggest that P120 and P140 could be analogous proteins in E. chaffeensis and E. canis.
Identification of ehrlichial P120, P140, P130, and P100 and steptococcal Fap1 based on their immunoreactivity clearly demonstrates that these glycoproteins are immunodominant (12, 15, 17,18). Immunoreactivity of the native and recombinant proteins is strong, and further investigation is needed to determine the role that carbohydrate has in immune recognition of these proteins. The recombinant E. chaffeensis P120 and E. canis P140 have demonstrated usefulness as serodiagnostic antigens (17,19). Consistent detection of antibodies in pre-convalescent-phase sera by using recombinant E. chaffeensis P120 (16) suggests that the attached glycans may elicit a T cell-independent immune response resulting in early production of antibody directed at these proteins.
We thank Thomas Bednarek for assistance with illustrations and Josie Ramirez-Kim for assistance in the preparation of the manuscript.
This study was supported by funding from the Clayton Foundation for Research and by a grant from the National Institute of Allergy and Infectious Diseases (AI31431).
Editor: J. T. Barbieri
Received 7 June 1999.
Returned for modification 27 August 1999.
Accepted 4 October 1999.
Beguin P.,
Miller J.,
Aubert J.-P.
(1993) Cellulose degradation by Clostridium thermocellum: from manure to molecular biology. FEMS Microbiol. Lett. 100:523–528.
Blattner F. R.,
Plunkett III G.,
Bloch C. A.,
Perna N. T.,
Burland V.,
Riley M.,
(1997) The complete genome sequence of Escherichia coli K12. Science 277:1453–1462.
Carl M.,
Dobson M. E.,
Ching W.-M.,
Dasch G. A.
(1990) Characterization of the gene encoding the protective paracrystalline-surface-layer protein of Rickettsia prowazekii: presence of a truncated identical homolog in Rickettsia typhi. Proc. Natl. Acad. Sci. USA 87:8237–8241.
Dobos K. M.,
Khoo K.,
Swiderek K.,
Brennan R.,
Belisle J. T.
(1996) Definition of the full extent of glycosylation of the 45-kilodalton glycoprotein of Mycobacterium tuberculosis. J. Bacteriol. 178:2498–2506.
Lechner J.
(1989) Structure and biosynthesis of prokaryotic glycoproteins. Annu. Rev. Biochem. 58:173–194.
Messner P.
(1997) Bacterial glycoproteins. Glycoconj. J. 14:3–11.
Moens S.,
Vanderleyden J.
(1997) Glycoproteins in prokaryotes. Arch. Microbiol. 168:169–175.
Nishimura H.,
Kawabata S.,
Kisiel W.,
Hase S.,
Ikenaka T.,
Takao T.,
Shimonishi Y.,
Iwanaga S.
(1989) Identification of a disaccharide (Xyl-Glc) and a trisaccharide (Xyl2-Glc) O-glycosidically linked to a serine residue in the first epidermal growth factor-like domain of human factors VII and IX and protein Z and bovine protein Z. J. Biol. Chem. 264:20320–20325.
Palmer E. L.,
Mallavia L. P.,
Tzianabos T.,
Obijeski J. F.
(1974) Electron microscopy of the cell wall of Rickettsia prowazekii. J. Bacteriol. 118:1158–1166.
9a.↵
Popov, V. L., X. J. Yu, and D. H. Walker. The 120-kDa outer membrane protein of Ehrlichia chaffeensis: preferential expression on dense-core cells and gene expression inEscherichia coli associated with attachment and entry. Microb. Pathog., in press.
Rikihisa Y.
(1991) The tribe Ehrlichieae and ehrlichial diseases. Clin. Microbiol. Rev. 4:286–308.
Sambi V.,
Stefanelli C.,
Cevenini R.
(1992) Detection of glycoproteins in Borrelia burgdorferi. Arch. Microbiol. 157:205–208.
Storey J. R.,
Doros-Richert L. A.,
Gingrich-Baker C.,
Munroe K.,
Mather T. N.,
Coughlin R. T.,
Beltz G. A.,
Murphy C. I.
(1998) Molecular cloning and sequencing of three granulocytic Ehrlichia genes encoding high-molecular-weight immunoreactive proteins. Infect. Immun. 66:1356–1363.
Tomoeda M.,
Inuzuka M.,
Date T.
(1975) Bacterial sex pili. Prog. Biophys. Mol. Biol. 30:23–56.
Weiss E.,
Coolbaugh J. C.,
Williams J. C.
(1975) Separation of viable Rickettsia typhi from yolk sac and L cell host components by renografin density gradient centrifugation. Appl. Microbiol. 30:456–463.
Wu H.,
Mintz P.,
Ladha M.,
Fives-Taylor P. M.
(1998) Isolation and characterization of Fap1, a fimbriae-associated adhesin of Streptococcus parasanguis FW213. Mol. Microbiol. 28:487–500.
Yu X. J.,
Crocquet-Valdes P.,
Cullman L. C.,
Popov V. L.,
Walker D. H.
(1999) Comparison of Ehrlichia chaffeensis recombinant proteins for diagnosis of human monocytotropic erhlichiosis. J. Clin. Microbiol. 37:2568–2575.
(1996) The recombinant 120-kilodalton protein of Ehrlichia chaffeensis, a potential diagnostic tool. J. Clin. Microbiol. 34:2853–2855.
(1997) Cloning and sequencing of the gene for a 120-kDa immunodominant protein of Ehrlichia chaffeensis. Gene 184:149–154.
Yu, X. J., J. W. McBride, C. M. Diaz, and D. H. Walker. Molecular cloning and characterization of the 120-kilodalton protein gene of Ehrlichia canis and serodiagnostic application for canine ehrlichiosis. J. Clin. Microbiol., in press.
Infection and Immunity Jan 2000, 68 (1) 13-18; DOI: 10.1128/IAI.68.1.13-18.2000
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Home OPINIONS Concerning the hijab
Concerning the hijab
by The Other Press October 21, 2015
Photo via theglobeandmail.com
Is the religious headwear oppression or expression?
By Aleeze Asif, Contributor
The hijab seems to have become a symbol of oppression worldwide. What is it about a scarf over the head that is seemingly oppressive compared to a scarf around the neck? The vast majority of the people that oppose the hijab seem to do so because of what they think it represents. So what exactly does it represent?
Islam dictates that all women must wear the hijab as a representation of their modesty and an expression of their religious identity (Quran 24:31). With that said, one must keep in mind that symbolic representation is fluid depending on the constantly evolving social and cultural circumstances, as well as the frame of mind of an individual.
It is an unfortunate reality that many Muslim women around the world are pressured into covering their heads by their families, cultural norms, or even the law. Modesty should be a choice, not another obligation put upon women because of their sex. A large portion of Muslim women, however, wear the hijab by choice. From an individualistic perspective, the reason why is quite simple: the hijab represents whatever the Hijabi wants it to represent. Many women wear it purely for the sake of religious obligation. Others say it incites confidence, and empowers them. Think about it: in a society where almost every conceivable media outlet is guilty of objectifying women at some point in time, who's to say the hijab isn't an act of rebellion?
However, things aren't as black and white when we analyze this religious practice from a broader sociological perspective. Islamic scripture and Hadith make it abundantly clear that the hijab is absolutely compulsory. The dictionary definition of discrimination is "different rules and standards for a sect of people established purely because of the way they are born." Gender-specific decorums are the reason why women might be the only group in the "other" category who aren't a minority. It wouldn't be inaccurate to argue that the hijab is guilty of promoting the further social segregation of genders by creating double standards and solidifying archaic gender norms. A woman should never have to do anything just because she is a woman.
Many countries are seeking to have the hijab banned. France is seeking to ban the headscarf in public places, having already banned any garment that covers a person's face, such as the niqab. Most Islamic scholars would testify to the fact that the niqab is not compulsory. Prime Minister Stephen Harper recently tried to ban the niqab, specifically at citizenship ceremonies, stating that it is "offensive" and it's "not how we do things here." There are legitimate arguments to restricting the niqab, as both Muslim and non-Muslim fanatics of either gender can use religious freedom as an excuse to hide their identities. However, it is profusely clear that that is not what Harper is concerned about. The fine line between expression and oppression is choice. Banning the niqab or hijab, however, is in itself a blatant violation of women's freedom of expression and a person's freedom of religion. How is telling women what they can't wear any better than telling them what they can wear?
The hijab might have its problems in the grand scheme of things, but reducing all Muslim women to a stereotype is tremendously counterproductive. There is a place for the hijab in the modern society that is pro-choice when it comes to matters of an individual's religious freedom. What a woman wears does not define her. It doesn't even begin to scratch the surface.
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Global Cleveland Kicks Off First Welcoming Week to Celebrate Immigrants
WKSU
The city of Cleveland is hosting its first Welcoming Week as Global Cleveland celebrates immigrants.
Today kicks off the first Welcoming Week in Cleveland. Hosted by Global Cleveland, the week offers a variety of events and special releases to celebrate immigrants in the community.
Joe Cimperman is the president of Global Cleveland. He says cities around the country have been doing these types of welcoming weeks, but this is the first one in Northeast Ohio.
"It's really part of a larger effort to highlight the incredible importance and impact that immigrants, newcomers, refugees have on our community," Cimperman says.
Global Cleveland lists the Welcoming Week Kick-off as happening around the city by showcasing global friendly employers. They have an interactive map on their website.
Global Cleveland kicked off the week by flying its flag in front of City Hall.
Credit GLOBAL CLEVELAND
GLOBAL CLEVELAND
Global Cleveland's flag flies alongside the American flag in front of City Hall.
On Tuesday, the organization will host a "Passport Party" at Market Garden Brewery. Attendees are asked to bring their passports and show off their stamps that could win them prizes. They'll also be able to receive a passport application, an application for renewal and a voter registration form.
Khizr Khan, the Gold Star father known for taking aim at then-candidate Trump during the 2016 Democratic National Convention, will also be in the city for a couple of events.
Khan is a constitutional lawyer who recently wrote a book about the constitution, This is our Constitution: What it is and Why it Matters. He'll be signing books at the Cleveland Public Library on Tuesday. Khan came to the U.S. from Pakistan and will be sharing his story during a forum at the City Club on Wednesday.
On Sunday, the Cleveland Art Museum will release a self-guided tour of immigrant artists and Global Cleveland will release a Spotify playlist for the week on Monday.
"We're gonna say to people, you know, get your week going with some really fun music from all over the world," Cimperman says. "Music from all over the world that's brought to you from people living here in Northeastern Ohio."
The week will end on Sept. 20 with a naturalization ceremony in Solon at the Cuyahoga County Public Library.
Cimperman says his mother was an immigrant from Slovenia and he thinks about her experience when doing outreach with the organization.
"My mom would have these moments of great loneliness and I think that's just part of the immigrant story in the United States," he says. "And I think events like this not only tell the world that we're open for business, that we welcome newcomers, that we want you to come here, but it says to the people who are already here how much we value them."
Other events throughout the week include an interfaith prayer service and luncheon with the United Church of Christ, a cocktail reception at Alexander Mann Solutions, and an informational breakfast about E-1/E-2 visas.
A full list of the weeks' events can be found on Global Cleveland's website. Some events require registration.
CommunityGlobal ClevelandCleveland immigrantsClevelandCity Club of ClevelandKhzir Khan
Cleveland To Host First Welcoming Week to Celebrate Immigrants
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See Prince Harry's Official Engagement Photos
Story from Royal Wedding 2018
Prince Harry & Meghan Markle's Engagement Photos Are Works Of Art
Photo: Chris Jackson/Getty Images.
They're officially engaged! The newest royal couple sealed their betrothal with three stunning engagement photos released on December 21. Kensington Palace tweeted the photos this morning. So far, the palace has released three photos, all of them taken at Frogmore House in Windsor earlier this week. The third photo is actually a "candid" shot, though it looks pretty professional to us.
"Prince Harry and Ms. Meghan Markle have chosen to release official photographs to mark their engagement," the Kensington Palace Twitter account wrote. The photos were taken by Alexi Lubomirski, a New York-based photographer known for his work with celebrities.
Prince Harry and Ms. Meghan Markle have chosen to release official photographs to mark their engagement.
— Kensington Palace (@KensingtonRoyal) December 21, 2017
As a way to say thank you, they have decided to share this candid photograph from the day of their portrait sittings directly with all of you. pic.twitter.com/MROyiKWEnG
All the photos show the couple in a somewhat intimate light — Markle and Harry, though as glamorous as ever, look as if they're sitting next to a wood-burning fire, sipping tea. (Royals can be casual, too!)
Markle and Harry announced their engagement in late November, sharing the news via the Prince of Wales' Twitter. Their wedding is actually fast approaching; the two will be married on May 19, 2018 at St. George's Chapel. After announcing their engagement, the couple gave a video interview, their first public interview as a couple. In the interview, Markle described how the actual question went down. According to her, the whole event was fairly casual.
"It was a cozy night. We were roasting chicken," Markle said of the proposal. "It was just an amazing surprise. It was so sweet and natural and very romantic. He got down on one knee... I could barely let [him] finish proposing! I said, 'Can I say yes now?'"
The two met on a blind date, a difficult feat for a royal prince to pull off. Nevertheless, the two hit it off, and shortly afterwards, Markle joined Harry for a camping trip in Botswana.
"I fell in love with Meghan so incredibly quickly — it was a confirmation to me that all the stars were aligned," Harry said in the interview. "This beautiful woman literally tripped and fell into my life."
He added that she's very good at handling all this "royal" stuff. "I fell into her life, and the fact that she will be unbelievably good at the job part of it as well is obviously a huge relief to me because she'll be able to deal with everything else that comes with it," he said.
Now, how do we get these photos made into wallpaper?
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If you've been following this blog recently, you've met Clover and Carrot.
As a quick update on Carrot, the rearing issue might have been resolved today with a 'knock it off' response to her intimidation tactics. Time will tell, but once outside, she immediately resorted to rearing again, but relented once she realized this tactic wasn't going to net anything but annoyance. It was a good test because there were a lot of horses in close proximity that could have occupied her full attention (usually the horses are in the barn or pastured further out when we train), yet she turned her full attention to the rider once she was discouraged from calling the shots.
When this little three-year-old half Arab filly trucked in, I thought she might rank up there with some of the most challenging Halcyon Acres' projects – including badly started horses that already had their brains seriously scrambled with early introductions to training and required considerable time and intuition to reach. LuLu underscored this conviction as she left a path of destruction in her wake – splintering boards as she kicked at horses in adjacent paddocks, jumping out of her paddock to go beat up other horses, breaking lead ropes and lunge lines and behaving in ways that made all the residents at the farm uncomfortable.
The day I entered the paddock without a cookie and was greeted with charging, teeth bared, kicking and striking is the day I decided to put her into training much earlier than planned. I like to give young horses a week or so to acclimate to the new surroundings and routine before engaging in a serious training regimen, but she earned an early start.
That first day was a knock-down drag-out battle of wills where merely asked her to move forward and/or stop with body language in the roundpen netted hostile responses. Ultimately, the goal was looking for her to acknowledge me, but we didn't quite get that far. We did manage to reach a point, though, where she allowed me to approach her as she stood quietly and politely.
The next day was a shocker (apparently she processed a lot more than she let on). I brought her to the roundpen and then set up a video camera, figuring this would be a good lesson in what's not working. She waited at the edge of the roundpen and watched as I set up the camera. I entered the fenced area, ignoring her and proceeding to the center. She trotted right up to me and stood. This filly had spent her life dominating all the animals and humans she encountered. She had never met a match that wasn't intimidated by her, yet didn't return her hostility in kind. After a day to think about all this, it seems she decided she liked the idea of having a leader she could respect from kind but insistent actions.
Today's challenge was the bridle. While she's been a bit uncomfortable with the bit in the past, she suddenly seemed terrified to have her mouth violated. This was honest fear – you could see it in her eyes. There's no way to know where this was coming from, but it was real. This filly's spent a lifetime being motivated by food and it's incredible how excited she gets about the mere sound of vittles. With this in mind, it seemed coating the bit with molasses would be an easy fix. That didn't work. What ultimately did was simply spending a good deal of time comforting her with the bit in front of her teeth, but not in her mouth. Once she calmed down and realized this wasn't going to hurt, she calmly and willingly accepted the bit. The rest of the lesson went great and we put her out with the big girls – where she's no longer queen bee, but seems to appreciate the reward of company over isolation.
We haven't had a problem with the bit since our first blow up. In fact, she now opens her mouth the moment it's presented (this gal is pretty oral anyway). She still has some issues with the girth being tightened, but if there's food in front of her, she's not bothered.
She became comfortable enough about the bit that we started long lining her and had our third session today. She's still a bit confused about the signals, but an exaggerated leading rein seemed to help her understand. It's kind of funny, as she seems to naturally gravitate toward an indirect rein, and her intended career is with Western tack.
Reading about some of these "difficult" horses makes me throw a big "thank you" at my horse every time–he's so good about so many things. I was cringing at the problem Lulu was having with taking the bit, while thinking about how my Billy can hardly wait to get the darn thing in his mouth! He goes fishing for it, like it's some kind of prize.
Thanks for stopping by and commenting, Alli. Billy sounds like a joy. LuLu actually is too, she just has some issues that we're working through. She wants to be good, and that helps a lot. Fortunatly (and this is very common with alphas), once we address a concern in a manner she can understand and accept, it doesn't usually surface again.


Fashion Week returns to New York Feb. 9 and now you can watch the shows streaming live online at http://www.youtube.com/liverunway.
Check out the latest trends for Spring and then hop on over to Lehigh Valley Mall to start updating your wardrobe!
Remember to catch the Lehigh Valley Mall Teen Board's Spring Fashion shows at 1pm and 3pm on Saturday, April 14th!
(Whitehall, PA, January 31, 2012) The Lehigh Valley Mall has chosen a new teen board for 2012. A total of thirty-five teens were selected from over three hundred and fifty applicants in the region.
The Lehigh Valley Mall Teen Board was established in 2001, for young men and women between the ages of thirteen to eighteen years, to represent Lehigh Valley Mall in fashion shows, special events and community service projects.
Now in its eleventh year, the program has evolved to offer local teens a voice in their communities, a way to contribute their energies in a positive manner, develop a better understanding and respect of each other's diverse backgrounds, and increase their own sense of self-worth and self-esteem through the positive impact they have on their own community.
Of the thirty-five members for 2012, twenty of the teens have returned from last year. Two of this year's members have now been with the program for five years and help to mentor new members. Over sixteen area middle and high schools are represented by this year's board.
The 2012 Teen Board members are Angela Arcelay, Greyson Asmus, Rowan Bienes, Fiona Bloise, Alecia Caballero, Amanda Childs, Lori Deutsch, Courtney Deutsch, Sierra Dreisbach, Grace Duah, Sofia Garra, Flannery Johnston, Tori Kraus, Samantha Lima, Morgan Linkhorst, Rebecca Lutterschmidt, Hope Newswanger, Jada Orchulli, Brittany Pantoni, Priyanka Patel, Keyli Peralta, Brianna Roberts, Rosemary Roberts, Kayla Rae Ruiz, Elizabeth Ruppert, Cara Schiavone, Judith Skinker, Lindsey Stout, Natasha Tax, Carlie Viemann, Alexis Yearde, Ashleigh Yearde, Abbi Walsh, Zuri Washington, and Holly Zapotocky.
You can look for the Teen Board at some of the upcoming events at the Lehigh Valley Mall, including a celebration of Dr. Seuss 75th Anniversary on February 25th from 11am-2pm; the arrival of the Easter Bunny on March 17th and of course the Teen Board Spring Fashion Show on April 14th at 1pm and 3pm.
Lehigh Valley Mall and Simon Youth Foundation will award one Community Scholarship valued at $1,400 to a deserving student from our local community.
The application period for the 2012 Simon Youth Foundation Community Scholarship awards is Tuesday, January 17, to Thursday, March 1.
You can learn more about eligibility requirements and download an application at www.syf.org. Applications also are available at Simon Guest Services.
Lehigh Valley Mall gets ready to select a new Teen Board for 2012!
The applications are all in! Over 375 teens applied for the 2012 program. This weekend, the semi-finalists selected will be interviewed, from which the final selections will be made and posted here.
Already, twenty-one members from 2011 have been already selected to return for another year. In addition, three teens, who volunteered with the program in 2011 were accepted for 2012.
Look for an announcement about our finalists before the month is over!
Teen Board members were on hand last night, Friday 11-11-11, to help usher in the holidays at Lehigh Valley Mall. Stationed at the Outdoor Shops to await Santa's arrival, they passed out free reindeer antlers and jingle bell necklaces to the myriad of children and families in attendance.
Teen Board members also staffed an information booth with holiday shopper guides as well as Kidgits information and activity sheets, helping shoppers get a head start on the holiday season.
It was a bit chilly out, but the mall was packed and everyone had a great time and started to get in to the holiday spirit!
Get Ready For Santa's Arrival!
Welcome Santa to the Lehigh Valley as the Whitehall Fire Department escorts him by Fire Truck to the Outdoor Shops. Children will enjoy reindeer ears to help show Santa the way to his Workshop. First, Santa will light the 25-foot Mama and 15-foot Baby Reindeer, and then parade the children to light the 30-foot Holiday Tree. He will finish his parade upon arrival to his Workshop, located in the Mall Event Court, where he stays for the Holiday season. The Radio Disney road crew will celebrate at Santa's Workshop with music, games and prizes at 7:00pm.
Teen Board will be on hand to hand out goodies and hot chocolate to the kids! Stop by and say hello – we will be at the Outdoor Shops Entrance in between LOFT and Barnes and Noble!


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Most Read: Contributor South Africa, December 2021
Kenya: The Proceeds Of Crime And Anti-Money Laundering (Amendment) Bill 2021
by Nigel Shaw and Jean Makaka
ENSafrica
The Proceeds of Crime and Anti-money Laundering Bill (the "Bill") gazetted on 03 September 2021 seeks to amend the current Proceeds of Crime and Anti-money Laundering Act, 2009 (the "Act"). The Bill proposes to introduce a number of changes targeted towards countering the current money laundering problem in Kenya.
The Bill provides for the removal of the position of deputy director of the financial reporting centre (the "Centre") in a bid to align the Act with the prevailing practice of not having deputy chief executives of state corporations.
The Bill expands the definition of "reporting persons" under the Act. This will now include: advocates, notaries and other independent legal professionals who are sole practitioners, partners or employees within professional firms so that the obligations to monitor complex unusual, suspicious, large or other transactions and to report any transactions that constitute or may be related to money laundering as provided under Part IV of the Act apply to them.
The Bill proposes that the obligations shall apply to the abovementioned parties while acting for their clients in the following circumstances:
buying and selling of real estate;
managing of client money, securities or other assets;
organisation of contributions for the creation, operation or management of companies; and
creation, operation or management of buying and selling of business entities.
In addition, the Bill seeks to give the Centre the authority to interrupt a transaction for not more than five working days where there is evidence of suspicious activity taking place. This will allow the Centre adequate time to investigate the transaction. The Bill also seeks to introduce provisions that will limit the right to privacy enshrined in the Constitution of Kenya, 2010 in relation to the prevention, detection and investigation of money laundering and financing of terrorism.
Further, the Bill proposes the introduction of the asset recovery oversight board (the Board). The function of the Board will include advising and overseeing the asset recovery agency (the Agency) on:
the exercise of its powers,
performance of its functions,
asset recovery policies,
strategic priorities and
the administration of the Agency.
The Board will hold the Agency accountable and will have the authority to constitute committees to ensure that this mandate is achieved.
The Bill is currently undergoing the legislative process and should be tabled for the first reading in parliament and thereafter public participation.
Kenya is considered one of the major money laundering jurisdictions across the globe. The Bill seeks to strengthen the current legislation around money laundering in the country by widening the scope of reporting requirements, addressing loopholes that have previously been used by money launderers in the past and is intended to improve vigilance in the financial sector. However, there is widespread concern as to the effect of the proposed amendments on several legal principles, practices and laws including client confidentiality, constitutional rights and criminal law principles and procedures.
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Jean Makaka
Kenya Government, Public Sector Constitutional & Administrative Law Money Laundering
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"I don't want to go to Nicaragua," I grumbled to my mom as I sat in the passenger's seat, wrinkling my nose. She had just asked me if I was excited about my new Peace Corps assignment. I still wasn't sure if I would actually go, but I said yes, for the moment. It was around New Year's Eve 2013, the end of a taxing year for both of us. It could only go uphill from here, I reminded myself.
We were driving our rental car through the surprisingly chilly New Mexican desert, a place that reminded me of Central Washington (where I grew up) because of its barren, beige-colored earth and open spaces. One thing was strikingly different: the cold, vast, cloudless, bright blue sky.
"Why don't you want to go?" asked my mom.
Concerned friends – who had never been to Nicaragua – warned me that I wouldn't be able to be myself there and that I should make the safer choice to stay in Boston. I grew up in a Mexican household, and I was never comfortable enough to come out to my mom until college. She was fine with it when I told her, but I wasn't comfortable to do so until I had met other queer people like myself. When you finally meet other people who understand you and where you're coming from, you become more comfortable with yourself.
Hearing my queer friends' successes and struggles with coming out to their friends and family inspired me to be more open about myself. In college, I not only met other queer women and transgender students, but most importantly to me, queer Latin@s. Leaving for Nicaragua meant losing the small network of queer Latin@s that took me so long to find, something I feared.
After six months in-country, I can safely say that Nicaragua has been amazing, to say the least. I feel as if I'm in the right place at exactly the right time in my life. When I Skype with my mom and friends back home, they can't help but comment on how relaxed I look and feel. "Man, and I thought you liked Boston but look at you now!" my friend said last night during a Skype date.
I told her that being here has made me realize how happy I am while I'm living abroad. I love waking up every day and facing the challenges and successes that come with navigating a different culture. I love that each day there is something to learn, whether it's learning to incorporate filler words like "Fijese de que…" or learning the proper way to write a cover letter in Spanish (hint: at the end, it is wise to say, "I wish you success in all of your daily activities" rather than the more American, "Thank you for your consideration").
In response to my previous concern that Nicaraguans would be a conservative, unwelcoming lot: false, false and false! While this is a highly religious country, Nicaraguans are some of the most generous, friendly people I have met. I've become used to the two typical questions I get asked: "Are you married?" and "Are you Catholic?" My response: no and yes.
I am lucky to live in a large, relatively progressive city in the mountains, so dealing with homophobia and other –isms isn't something I worry about as much as I would in a smaller community. There are lots of NGOs, women's collectives, queer Latin@s and social justice work going on. I feel at home here because there's a strong sense of people helping one another out, regardless of what they have.
As a TEFL teacher trainer, my job is to co-teach high school English classes with Nicaraguan teachers. I also lead teaching workshops to other Peace Corps Volunteers and staff, and teach English classes for teachers and community members. I've had plenty of time to integrate into my community, cook chilaquiles, teach English, eat pastel cuatro leches and go hiking. I'm looking forward to working with Fundación Uno, which sponsors weekend English classes for Nicaraguan English teachers. Doing these trainings will be a top priority for me, since my goal is to empower Nicaraguan English teachers. They will be the ones who will stay when and if I ever leave.
I can't imagine being anywhere else at the moment, and am looking forward to overcoming life's obstacles with more patience and optimism than ever. I've been lucky enough to find my queer Latin@ community in Nicaragua, and am grateful every day for it. For now, I don't want to leave Nicaragua.
Charleen Johnson Stoever is a 24-year-old Mexican American queer woman who is serving as a TEFL Peace Corps Volunteer in Nicaragua (2014-16). She earned a B.A. in French and Women's Studies at Wellesley College. She believes in the power of education as a tool for social mobility.


3. Insertion of new section 78A in Act 1 of 1872.
An Act to amend the Indian Evidence Act, 1872 in its application to West Bengal.
1. Short title and extent. - (1) This Act may be called the Indian Evidence (West Bengal Amendment) Act, 1955.
2. Application of Act. - The Indian Evidence Act, 1872 (hereinafter referred to as the said Act), shall, in its application to West Bengal, be amended for the purpose and in the manner hereinafter provided.
"78A. Copies of public documents to be as good as the original documents in certain cases. - Notwithstanding anything contained elsewhere in this Act or in any other law for the time being in force, where any public documents concerning any areas within a partitioned district or subdistrict have been kept in East Bengal, then copies of such public documents shall, on being authenticated in such manner as may be prescribed from time to time by the State Government by notification in the Official Gazette, be deemed to have taken the place of, and to be, the original documents from which such copies w6re made and all references to the original documents shall be construed as including references to such copies.
(2) "East Bengal" means the territories now comprised in the Province of East Bengal forming part of the Dominion of Pakistan.".


text:
Chemical Engineering and Analytical Science
Mechanical, Aerospace and Civil Engineering
It's my pleasure to provide an introduction to Edition 06 of In Abstract. For your reading pleasure, our expert panel have selected highlights of some of the most spectacular world-leading research advances recently made by researchers in the Faculty of Science and Engineering at The University of Manchester.
As has been remarked upon in previous editions, these advances are as diverse as they are exciting, from the development of 4D techniques to study geological processes in hot magma, to novel catalysts for the efficient synthesis of important drug molecules, to new insights into assessing the reproducibility of important algorithms, and beyond!
Nitrogen dioxide is a major pollutant from exhausts that causes significant environmental and health issues, and chemists at Manchester have shown for the first time that this gas can be selectively and fully reversibly stored in a class of porous materials. Manchester chemists have also used rapid genome evolution of yeast cells to fast-track the development of biological engineering.
The School of Materials showcases diversity in contemporary materials science, by investigating the response of major players in the fashion and textile industry to modern slavery legislation, the development of wear-resistant materials for application in nuclear reactors, and a collaboration with the School of Physics and Astronomy to investigate gas flow dynamics in 2D materials. Also within Physics, forget what you have previously learned about electricity and water not mixing – a new measurement has proved that whilst this is true for bulk water, its surface remarkably shows no interaction with an electric field.
Researchers in Computer Science have modelled the connectivity of neurones and synapses to simulate processes in a small portion of the human brain, whilst in Electrical and Electronic Engineering a team has helped develop novel self-powered wireless sensors; these devices have a plethora of potential applications in security and health monitoring.
Finally, an additional highlight for those who don't suffer from arachnophobia is provided by a massive leap in our understanding of jumping spiders, which has given new inspiration for the design of agile micro-robots.
Simply click on the individual abstract titles below to find out more about all these remarkable research studies and many others. The next edition of In Abstract will be published in a few months, so please do visit this site again in the near future to catch up on the next wave of Manchester-led science and engineering advances.
Horizontal Collaboration in Response to Modern Slavery Legislation: An Action Research Project


text:
3 Reasons Why Apple's CEO Is Not Worried About Falling iPhone Sales
By Andrs CardenalFool.com
Apple stock is under considerable selling pressure lately. Shares of the tech juggernaut are down by more than 28% in the last year due to falling revenue in the very-important iPhone segment. However, CEO Tim Cook believes that the decline in iPhone sales is only a short-term problem.
Nearly 65% of total Apple revenue comes from the iPhone segment, so the product is absolutely crucial in terms of overall financial performance. Apple registered a 16% decline in iPhone unit sales during the quarter ended in March, and total company-level revenue fell 13% versus the same quarter in 2015.
But that's old news for investors at this stage. The big question to consider is whether the decline is only temporary, or if Apple is on a long-term decline due to languishing iPhone demand in the years ahead.
Apple CEO Tim Cook is an interested party in this discussion, so investors need to take some distance when analyzing his point of view. Nevertheless, Mr. Cook has some strong arguments to believe that the iPhone will continue growing in the future. Here are his own words, from Apple's last earnings conference call:
The iPhone 6 was an explosive success for Apple in 2015. Many customers were eagerly waiting for a new iPhone with a larger screen, so sales skyrocketed. The iPhone 6s is not a big game changer in comparison to previous models, and this is making comparisons remarkably challenging in 2016.
The company sold 61.2 million devices during the quarter ended in March of 2015, a huge increase of 40% versus the same quarter in 2014. Unit sales during the quarter ended in March of 2016 amounted to almost 51.2 million. The number is down by 16% versus the same quarter in 2015, but still up by 17% versus 2014 levels.
This is important to consider because it indicates that the decline in iPhone sales this year could be mostly due to tough comparisons against 2015 levels. Besides, Apple customers are remarkably loyal to the brand, and a gargantuan installed based bodes well in terms of renewals going forward.
According to Tim Cook:
Switchers from Android
According to IDC numbers, Apple has a global market share of 15.2%, while a huge 82.6% of all smartphones around the world work with Android. This means that, even if overall industry growth is slowing down, Apple still has a lot of room to steal customers away from the competition. Tim Cook said in the last conference call that Apple is attracting record amounts of switchers from Android and other platforms, and this could be an interesting growth driver going forward.
Growing smartphone penetration
The smartphone market in the U.S. and other developed countries is highly saturated, but things are remarkably different in emerging markets, where smartphone penetration levels are still quite low. Income levels in many of these countries are also much lower, so it's hard to tell what levels of market share Apple can obtain in these markets over the middle term. However, that's no reason to discard growth opportunities due to increasing smartphone penetration in emerging markets.
In fact, Apple has recently launched its new iPhone SE model, which sells for a comparatively low starting price of $399 versus a much-higher starting price of $649 for the iPhone 6s. This device seems to be particularly targeted toward capitalizing on growth opportunities in emerging markets.
According to Tim Cook, growing smartphone penetration should generate considerable opportunities for expansion in key emerging markets such as India.
Only time will tell for certain if Mr. Cook is right or wrong about growth prospects in the iPhone segment. However, between first-time smartphone buyers, customers switching away from other platforms, and renewal sales to current iPhone users, it looks like Apple's iconic product still has room for growth.
The article 3 Reasons Why Apple's CEO Is Not Worried About Falling iPhone Sales originally appeared on Fool.com.
Andrs Cardenal owns shares of Apple. The Motley Fool owns shares of and recommends Apple. The Motley Fool has the following options: long January 2018 $90 calls on Apple and short January 2018 $95 calls on Apple. Try any of our Foolish newsletter services free for 30 days. We Fools may not all hold the same opinions, but we all believe that considering a diverse range of insights makes us better investors. The Motley Fool has a disclosure policy.


text:
1969 US Open: Rod Laver's road to the final
When I strode onto the court at Forest Hills, New York, in the first round of the US Open in the last week of August, I had won 23 matches in a row, beginning with my first round victory over Nicola Pietrangeli at Wimbledon. The last person to beat me was John Newcombe at Queen's three months earlier. I had been fretting that my winning streak, one that was unprecedented in my career, would come to an end, as purple patches always do, and that when my luck ran out it would be at the US Open at the West Side Tennis Club at Forest Hills. […]
When I entered the cauldron to play against Mexican Luis Garcia in the first match of the Open, I was at peace with myself. I beat young Garcia, and then the Chileans Jaime Pinto-Bravo and Jaime Fillol without dropping a set.
Dennis Ralston, whom I faced in my fourth match, was a different proposition, a living, breathing wake-up call that I was not going to win this tournament without a mighty struggle. Dennis, who had pressed me in the past but was yet to beat me, lost the first set and then, playing the best tennis I had ever seen from him, won the next two. The 10,000-strong crowd at Forest Hills that gloriously sunny afternoon forgave their countryman for some disappointing Davis Cup performances in the past, and cheered him passionately. Their cheers were most ecstatic when he walked to the baseline after we'd had a breather when we changed ends, and when I stepped up there'd be a deafening silence and baleful glares that left me in no doubt that I wore the black hat that day. So to neutralise the crowd's support for Dennis, I fell back on my old Harry Hopman ploy and assumed my position at the baseline at precisely the same time Dennis did. That way I soaked up the cheers that were meant for him, and nobody was going to boo for fear that Ralston would think he was the one copping the Bronx cheers.
I downed Dennis in five sets, 6-4 4-6 4-6 6-2 6-3. He succumbed to my persistent pressure in the fourth set and his hangdog body language in the fifth told me that his gallant resistance was finished and I overpowered him.
We Aussies stuck together and Emmo and Fred Stolle played a part in my win over Dennis when they came to me in the dressing room during the 10-minute break after the third set and told me to toss the ball up a little higher when I served, because many of my serves that day had been finding the net. Their advice paid dividends.
After my match with Dennis, the heavens opened over New York and in 48 hours dumped 16 centimetres of rain. The courts of the West Side Tennis Club became a quagmire. The organisers of this prestige tournament were ill-prepared. The few flimsy tarpaulins they had handy were never going to protect the court surfaces, which, because of the sparse covering of grass over their soil, turned to viscous mud and were not remotely playable. For the next two and a half days, while we waited for the deluge to end, I practised on indoor courts, worked out and had soothing hot Epsom salts baths in the gymnasium of the New York athletic club.
In my quarter final against my old mate and nemesis of so many years, Roy Emerson, my rhythm was out of whack and I began sluggishly. Before I knew it, Emmo had won the first set 6-4 on a court that was still waterlogged, which made the balls heavy and sodden. Then Emmo broke my serve early in the second set. I concentrated on settling myself and slowing down, doing the little things right, like keeping my eye on the ball and hitting through it, breathing evenly, getting my serves in by placing them precisely, simply returning back over the net anything he hit at me without necessarily going for winners, punishing his second serve… all the tried and tested stuff.
As slippery as that court was, I was moving quickly, faster than Emmo, and battling my way back into the match. I won the second set 8-6, and the third 13-11 on the back of a little good luck. Roy hit a forehand passing shot that he believed was good but which the linesman ruled out. Deuce. I then produced two passing forehands that he was unable to handle to win the set. In the end, it was the court as much as me that ended his US Open. He had a way of dragging his foot when he served and he was chewing up the baseline so much that soon there was no solid section of the court from which to serve. Then he began catching his dragging foot in the sludge. When it was my turn to serve from the end Roy had ripped up, it was not a problem for me for the simple reason that I did not drag my foot. I was ahead 5-4 in the fourth and serving for the match when I surprised Emmo with my first top-spin lob of our encounter. He was racing in to the net when the ball arced back over his head. The slippery court prevented Roy from stopping and turning to give chase, and he didn't even try – he just kept running to the net, his hand outstretched and grinning that big gold-toothed grin of his.
On his way to our semi final showdown, Arthur Ashe's sledgehammer serve had seen him convincingly beat Manuel Santana and Muscles Rosewall, so he was on a roll. And of course in our match the crowd would be right behind him. With a final depending on the result, I needed no added incentive to beat Arthur, but I had a point to prove against him, because he had been putting it about that it was time for older generation players such as Muscles and me to stand aside for him and his contemporaries Newk, Rochey and Tom Okker. Speaking for myself, I had no intention of vacating the scene for anybody just yet, and I was determined to show Arthur that there was life in this old dog yet.
After I came onto court at the same time as him to bask in the cheers of his supporters, the Americans who made up the vast majority of the crowd, he went immediately into top gear, much as he had done at Wimbledon. His dynamic serves had me on the defensive and, at 5-4 to him, he served for the first set. Then Arthur's inexperience brought him undone. Instead of coolly and accurately placing a winning serve, he attempted to ace me and the ball sailed out. His softer second serve was easy meat. I did my usual thing of just returning the ball to him firmly but surely, sending back to him everything he hit at me, applying the pressure back on him to hit winners. I was chipping, straight down the middle of the court and depriving him of the wide angles he needed to slam winners.
The first set was mine 8–6, and then I won the second decisively, 6-3. In the third, however, Arthur gave a glimpse of the skill and tenacity that were to make him a far better player in years to come than he was in 1969. After he led 3-0, I caught him up and we remained locked together on serve until night fell and play was called off for the day with the score 12-12. We left the court lost in our thoughts. I got the impression from Arthur's worried expression that he would not be sleeping well that night. Arthur knew what he had to do; wrap up the third set the next morning and take the match to five sets. And I knew what he had to do, so I planned to blitz him from the outset, win the next two games, the fourth set, and the match. […]
When I did put my head on the pillow sometime after 11, I slept like a baby and felt fresh, relaxed and ready for action when Arthur, who looked drawn, and I resumed our match next morning. Also in my favor was that the was serving first. Now, serving after a break or first in a new match is never easy because you've had no time to warm up and get your arm and eye in. This is why I chose to receive when I won the toss in a match – it gave me a good chance of breaking my opponent's serve. Which is what I did, and I held my own serve to wrap up those two games and win the match in straight sets. I was in the final.
That was the good news. The bad was that I'd be playing Tony Roche, who was in tremendous form and dead keen to avenge his loss to me in the Australian Open semi final in January on the back of what he will believe to his dying day was a questionable call.
I couldn't help thinking of the parallels between me and tony, who was, and remains, a good friend: he hailed from a tiny Australian country town like me, and his dad had also been a butcher. I thought again of how Ken Rosewall scuttled his mate and doubles partner Lew Hoad's Grand Slam bid in the US championships in 1956. My fear that my friend Roy Emerson would do the same to me in the final of the 1962 US championships proved groundless, but now I wondered whether it was ordained that this time another mate would end my Grand Slam bid. Then I banished those qualms from my mind. This match would be won by the better player on the day and anything that happened 13 or even 7 years ago would have no bearing.
Photo by Walter Iooss Jr. /Sports Illustrated/Getty Images
– 1969 US Open: Rod Laver completes his second Grand Slam
Filed under Books, US Open · Tagged with 1956, 1962, 1969, A Memoir, Arthur Ashe, Dennis Ralston, Fred Stolle, Harry Hopman, Jaime Fillol, Jaime Pinto-Bravo, John Newcombe, Ken Rosewall, Lew Hoad, Luis Garcia, Manuel Santana, Nicola Pietrangeli, Rod Laver, Roy Emerson, Tom Okker, Tony Roche, usopen1956, usopen1962, usopen1969
Wimbledon 2017 coverage
Three weeks after the victories of Jelena Ostapenko and Rafael Nadal at Roland Garros, all players have their eyes turned to the grass courts of Wimbledon. With the absences of Serena Williams and Maria Sharapova, the women's draw is once again wide open, while Roger Federer is the big favorite for the title in the men's draw.
Follow our coverage on Tennis Buzz and leave us a comment if you want to share your pictures and stories.
Fan's guide:
How to get Wimbledon tickets
wimbledon2017tickets
How to get last minute Wimbledon tickets
27939953982_27b9e00190_z
Wimbledon Queue 101
queue_wimbledon
Tennis culture: Wimbledon victory climb
murray_climb_wimbledon2013
A trip down memory lane:
Wimbledon memories: Mrs Blanche Bingley Hillyard
Wimbledon memories: Charlotte Cooper Sterry
Wimbledon memories: Dora Boothby
Portrait of Wimbledon champion Ann Jones
Wimbledon 1969: Laver's getting beat by an Indian
Rod Laver – John Newcombe Wimbledon 1969
Around the grounds at Wimbledon in 1971
Wimbledon 1975: Ashe vs Connors
1976: Bjorn Borg first Wimbledon title
Portrait of 5-time Wimbledon champion Bjorn Borg
Wimbledon 1976: Chris Evert defeats Evonne Goolagong
Portrait of Virginia Wade, winner in 1977
Wimbledon 1978 in pictures
1978: First Wimbledon title for Martina Navratilova
1978: Bjorn Borg defeats Jimmy Connors
Wimbledon 1979: Passing on the record
1981: First Wimbledon title for McEnroe
1982: Jimmy Connors defeats John McEnroe
1984: John McEnroe defeats Jimmy Connors
1985: Boris Becker, the man on the moon
1986: Boris Becker defeats Ivan Lendl, wins second Wimbledon title
Portrait of 3-time Wimbledon champion Boris Becker
Wimbledon 1987 SF Cash defeats Connors
Wimbledon 1987 Cash defeats Lendl
Wimbledon 1988: An era ends as Graf beats Navratilova
Wimbledon 1988: Edberg a deserving new champion
Portrait of 2-time Wimbledon champion Stefan Edberg
Wimbledon 1990: Becker vs Edberg
1990: Martina Navratilova's historic 9th Wimbledon title
Wimbledon 1991: the first Middle Sunday
1991: Michael Stich defeats Boris Becker
1992: first Grand Slam for Andre Agassi
Andre Agassi: thanks to Wimbledon I realized my dreams
1993: Pete Sampras defeats Jim Courier
1994: Pete Sampras defeats Goran Ivanisevic
1995: Tim Henman disqualified!
Wimbledon 1996: singing in the rain
1996: Richard Krajicek upsets Pete Sampras
Wimbledon 1996: a winning streak
1997: Pete Sampras defeats Cédric Pioline
2000 Wimbledon SF: Pat Rafter defeats Andre Agassi
Wimbledon 2000: did dad call the shots?
2000 Wimbledon Final: Pete Sampras defeats Pat Rafter
2001 Wimbledon 4th round: Federer defeats Sampras
Wimbledon 2001 People's Final: Ivanisevic vs Rafter
Wimbledon 2010: Rafael Nadal defeats Tomas Berdych
Wimbledon 2012: Roger Federer defeats Andy Murray
Andy Murray's road to the Wimbledon 2013 final
Wimbledon 2013: Andy Murray, 77 years after Fred Perry
Discuss:
What if Edberg had coached Henman?
Fashion and gear:
Wimbledon 2017: Garbine Muguruza outfit
muguruza_adidas_wimbledon2017
Wimbledon 2017: Dominic Thiem and Sascha Zverev's outfits
thiem4_adidas_wimbledon2017
Wimbledon 2017: Kerber, Halep and Mladenovic outfits
Wimbledon 2017: Kei Nishikori Uniqlo outfit
nishikori_uniqlo_wimbledon2017
Nike celebrates Federer's 8th Wimbledon title
Who will win Wimbledon 2017?
Karolina Pliskova (19%, 4 Votes)
Venus Williams (19%, 4 Votes)
Angelique Kerber (14%, 3 Votes)
Simona Halep (14%, 3 Votes)
Johanna Konta (14%, 3 Votes)
Someone else (10%, 2 Votes)
Svetlana Kuznetsova (5%, 1 Votes)
Agnieszka Radwanska (5%, 1 Votes)
Elina Svitolina (0%, 0 Votes)
Dominika Cibulkova (0%, 0 Votes)
Caroline Wozniacki (0%, 0 Votes)
Filed under Books, Fashion, Gear, Slider, Wimbledon · Tagged with 1969, 1971, 1975, 1976, 1977, 1978, 1981, 1982, 1984, 1985, 1986, 1987, 1988, 1990, 1991, 1992, 1993, 1994, 1995, 1996, 1997, 2000, 2001, 2012, 2013, 2014, 2015, 2016, 2017, adidas, Alexander Zverev, Andre Agassi, Andy Murray, Ann Jones, Arthur Ashe, Bjorn Borg, Blanche Bingley Hillyard, Boris Becker, Cedric Pioline, Charlotte Cooper Sterry, Chris Evert, Dominic Thiem, Dora Boothby, Evonne Goolagong, Goran Ivanisevic, Ilie Nastase, Ivan Lendl, Jim Courier, Jimmy Connors, John McEnroe, John Newcombe, Kei Nishikori, Malivai Washington, Martina Navratilova, Michael Stich, Novak Djokovic, Pat Cash, Pat Rafter, Pete Sampras, Premjit Lall, Rafael Nadal, Richard Krajicek, Rod Laver, Roger Federer, Serena Williams, Stefan Edberg, Steffi Graf, Tim Henman, Tomas Berdych, Uniqlo, Venus Williams, Virginia Wade, wb1969, wb1971, wb1975, wb1976, wb1977, wb1978, wb1981, wb1982, wb1984, wb1985, wb1986, wb1987, wb1988, wb1990, wb1991, wb1992, wb1993, wb1994, wb1995, wb1996, wb1997, wb2000, wb2001, wb2012, wb2013, wb2014, wb2015, wb2016, wb2017, Wimbledon Museum
The All England Lawn Tennis and Croquet Club:
Wimbledon guided tour – part 1
Wimbledon Centre Court roof
Court 3 : a new Show Court at Wimbledon
Waiting in the Queue to Wimbledon
Wimbledon Museum: The Queue exhibition
The Wimbledon Lawn Tennis Museum: Player Memorabilia
Wimbledon 's biggest upsets
1990: Martina Navatilova's historic 9th Wimbledon title
The Spirit of Wimbledon: a 4-part documentary by Rolex retracing Wimbledon history
Ralph Lauren unveils new Wimbledon uniforms
ralphlauren1_wimb2016
Wimbledon 2016: Djokovic and Nishikori Uniqlo outfits
nishikori_wimbledon2016
Wimbledon 2016: Kerber, Berdych and Zverev outfits
kerber_adidas_wimbledon2016
Wimbledon 2016: Serena, Federer and Kyrgios Nike outfits
federer_nike_wimbledon2016
Novak Djokovic (53%, 50 Votes)
Roger Federer (21%, 20 Votes)
Andy Murray (17%, 16 Votes)
Dominic Thiem (5%, 5 Votes)
Stan Wawrinka (1%, 1 Votes)
Kei Nishikori (1%, 1 Votes)
Tomas Berdych (1%, 1 Votes)
Milos Raonic (1%, 1 Votes)
Richard Gasquet (0%, 0 Votes)
David Goffin (0%, 0 Votes)
Someone else (0%, 0 Votes)
Serena Williams (33%, 8 Votes)
Garbine Muguruza (33%, 8 Votes)
Victoria Azarenka (17%, 4 Votes)
Simona Halep (8%, 2 Votes)
Angelique Kerber (4%, 1 Votes)
Roberta Vinci (0%, 0 Votes)
Belinda Bencic (0%, 0 Votes)
Venus Williams (0%, 0 Votes)
Timea Bacsinszky (0%, 0 Votes)
Filed under Books, Fashion, Gear, Slider, Wimbledon · Tagged with 1969, 1971, 1975, 1976, 1977, 1981, 1982, 1984, 1985, 1986, 1988, 1990, 1992, 1993, 1994, 1995, 1996, 1997, 2000, 2001, 2012, 2013, 2014, 2015, 2016, adidas, Andre Agassi, Andy Murray, Ann Jones, Arthur Ashe, Bjorn Borg, Blanche Bingley Hillyard, Boris Becker, Cedric Pioline, Charlotte Cooper Sterry, Chris Evert, Dora Boothby, Evonne Goolagong, Goran Ivanisevic, H&M, Ilie Nastase, Ivan Lendl, Jim Courier, Jimmy Connors, John McEnroe, John Newcombe, Kei Nishikori, Lacoste, Laura Robson, Malivai Washington, Martina Navratilova, Milos Raonic, New Balance, Nick Kyrgios, Nike, Novak Djokovic, Pat Rafter, Pete Sampras, Premjit Lall, Rafael Nadal, Richard Krajicek, Rod Laver, Roger Federer, Stefan Edberg, Steffi Graf, Tim Henman, Tomas Berdych, Under Armour, Uniqlo, Virginia Wade, wb1969, wb1971, wb1975, wb1976, wb1977, wb1981, wb1982, wb1984, wb1985, wb1986, wb1988, wb1990, wb1992, wb1993, wb1994, wb1995, wb1996, wb1997, wb2000, wb2001, wb2012, wb2013, wb2014, wb2015, wb2016, Wimbledon Museum
The Rocket Rod Laver
From Love Thirty: Three Decades of Champions, by Rex Bellamy, published in 1990:
Rodney George Laver was the most astounding player I ever saw, and may have been the greatest ever. His record is without parallel. Consider what that record might have been but for his exclusion from 21 Grand Slam tournaments when he was, presumably, at his physical peak, between the ages of 24 and 29. Had professionals been eligible for those events, Lew Hoad might have had the better of laver for a year or so and Ken Rosewall would always have been worth an even-money bet. But one has to believe that from 1963 to 1967 Laver would have collected another bunch of major championships and perhaps a third Grand Slam. Laver overlapped and dominated two Grand Slam eras separated by seven years. He did so because he had it all. Because he was adventurer and artist in one. Because he could raise his game to any level demanded of it.
Laver was only 5ft 8 1/2in tall and usually weighed around 10st 71lb. But he had gigantic left arm and his speed and agility were breathtaking. The circumference of his left forearm was 12in and the wrist measured 7in. The strength of that wrist and forearm gave him blazing power without loss of control, even when he was on the run at full stretch. The combination of speed and strength, especially wrist-strength, enabled him to hit ferocious winners when way out of court – often when almost under the noses of the front ow of spectators. And he was a bow-legged, beautifully balanced, and as quick as a cat. He had some glorious matches with Rosewall – and with Tom Okker, who could match Laver's speed and panache but was second-best in terms of strength and technical versatility. Laver also had the eyes of a hawk and fast anticipation and reactions. Like Budge, he was feckle-faced and had copper-coloured hair. Another distinguished feature was a long nose that, in spite of the kink in it, gave a false impression of hauteur. For much of his career Laver was confessedly shy and self-conscious, but there was no 'side' to him. He was easy going – except on court.
Marty Riessen once summed up Laver admirably: "To look at him walking around, you wouldn't think he was world champion. He doesn't stand out. His stature isn't something you expect, like a Gonzales or a Hoad. Off the court, his personality seems almost retiring. But it's as if he goes into a telephone booth and changes. On court he's aggressive. Such a big change of personality – when a lot of players play the same as they act. What impresses me is his quickness. Speed enables him to recover when he's in trouble. And the thing I learned from playing Laver is how consistent one can be with power. It's amazing how he can keep hitting with such accuracy. He combines everything. There are a lot of good competitors. But he's fantastic."
Filed under ATP, Books, Davis Cup, Players, US Open · Tagged with 1951, 1952, 1956, 1959, 1960, 1962, 1968, 1969, 1971, 1972, 1973, André Licis, Arthur Ashe, Arthur Drysdale, Australia, Bjorn Borg, Bob Mark, Brisbane, Charlie Hollis, Cleveland, Cliff Drysdale, Dallas, daviscup1973, Dennis Ralston, Dick Crealy, Donald Budge, Dunlop, Forest Hills, Harry Hopman, Ion Tiriac, John Newcombe, Ken Rosewall, Lew Hoad, Martin Mulligan, Marty Riessen, Mary Laver, Neale Fraser, ozopen1969, Premjit Lall, Rex Bellamy, rg1969, Rockhampton, Rod Laver, Roger Taylor, Roy Emerson, Stan Smith, Tom Okker, Tony Roche, United States, usopen1969, wb1962, wb1969, WCT, WCT1972


text:
<?php

namespace Blast\Tests\Orm\Stubs\Definition;

use Blast\Orm\Entity\Definition;

class ProjectDefinition extends Definition
{

    /**
     * ProjectDefinition constructor.
     */
    public function __construct()
    {
        $this->setConfiguration([
           'primaryKeyName' => 'uid'
        ]);
    }
}

redpajama_set_name: RedPajamaGithub