123 Hits in 0.97 sec

Structure of the Induced Antibacterial Protein from Tasar Silkworm,Antheraea mylitta

Deepti Jain, Deepak T. Nair, G. Jawahar Swaminathan, E. G. Abraham, J. Nagaraju, Dinakar M. Salunke
2001 Journal of Biological Chemistry  
The crystal structure of an antibacterial protein of immune origin (TSWAB), purified from tasar silkworm (Antheraea mylitta) larvae after induction by Escherichia coli infection, has been determined. This is the first insect lysozyme structure and represents induced lysozymes of innate immunity. The core structure of TSWAB is similar to c-type lysozymes and ␣-lactalbumins. However, TSWAB shows significant differences with respect to the other two proteins in the exposed loop regions. The
more » ... ic residues in TSWAB are conserved with respect to the chicken lysozyme, indicating a common mechanism of action. However, differences in the noncatalytic residues in the substrate binding groove imply subtle differences in the specificity and the level of activity. Thus, conformational differences between TSWAB and chicken lysozyme exist, whereas functional mechanisms appear to be similar. On the other hand, ␣-lactalbumins and c-type lysozymes exhibit drastically different functions with conserved molecular conformation. It is evident that a common molecular scaffold is exploited in the three enzymes for apparently different physiological roles. It can be inferred on the basis of the structure-function comparison of these three proteins having common phylogenetic origin that the conformational changes in a protein are minimal during rapid evolution as compared with those in the normal course of evolution.
doi:10.1074/jbc.m104674200 pmid:11522783 fatcat:zckpdselvjct3fmkjls2mmj3by

Mapping the Ribonucleolytic Active Site of Eosinophil-derived Neurotoxin (EDN)

Demetres D. Leonidas, Ester Boix, Robert Prill, Motoshi Suzuki, Richard Turton, Kathryn Minson, G. Jawahar Swaminathan, Richard J. Youle, K. Ravi Acharya
2001 Journal of Biological Chemistry  
Eosinophil-derived neurotoxin (EDN), a basic ribonuclease found in the large specific granules of eosinophils, belongs to the pancreatic RNase A family. Although its physiological function is still unclear, it has been shown that EDN is a neurotoxin capable of inducing the Gordon phenomenon in rabbits. EDN is also a potent helminthotoxin and can mediate antiviral activity of eosinophils against isolated virions of the respiratory syncytial virus. EDN is a catalytically efficient RNase sharing
more » ... milar substrate specificity with pancreatic RNase A with its ribonucleolytic activity being absolutely essential for its neurotoxic, helminthotoxic, and antiviral activities. The crystal structure of recombinant human EDN in the unliganded form has been determined previously (Mosimann, S. C., Newton, D. L., Youle, R. J., and James, M. N. G. (1996) J. Mol. Biol. 260, 540 -552). We have now determined high resolution (1.8 Å) crystal structures for EDN in complex with adenosine-3,5-diphosphate (3,5-ADP), adenosine-2,5-diphosphate (2,5-ADP), adenosine-5-diphosphate (5-ADP) as well as for a native structure in the presence of sulfate refined at 1.6 Å. The inhibition constant of these mononucleotides for EDN has been determined. The structures present the first detailed picture of differences between EDN and RNase A in substrate recognition at the ribonucleolytic active site. They also provide a starting point for the design of tight-binding inhibitors, which may be used to restrain the RNase activity of EDN.
doi:10.1074/jbc.m010585200 pmid:11154698 fatcat:hc6w43u6yzhblndf2opvzpdqvm

Crystal Structure of the Eosinophil Major Basic Protein at 1.8 Å

G. Jawahar Swaminathan, Arthur J. Weaver, David A. Loegering, James L. Checkel, Demetres D. Leonidas, Gerald J. Gleich, K. Ravi Acharya
2001 Journal of Biological Chemistry  
The eosinophil major basic protein (EMBP) is the predominant constituent of the crystalline core of the eosinophil primary granule. EMBP is directly implicated in epithelial cell damage, exfoliation, and bronchospasm in allergic diseases such as asthma. Here we report the crystal structure of EMBP at 1.8 Å resolution, and show that it is similar to that of members of the C-type lectin superfamily with which it shares minimal amino acid sequence identity (ϳ15-28%). However, this protein lacks a
more » ... a 2؉ /carbohydrate-binding site. Our analysis suggests that EMBP specifically binds heparin. Based on our results, we propose a possible new function for this protein, which is likely to have implications for EMBP function.
doi:10.1074/jbc.m100848200 pmid:11319227 fatcat:yl6qhtw6rvhpfkjmd6mazj6hhe

Household Contact Screening and Yield of Tuberculosis Cases—A Clinic Based Study in Chennai, South India

Dina Nair, Nandita Rajshekhar, Joel Shyam Klinton, Basilea Watson, Banurekha Velayutham, Jaya Prasad Tripathy, Mohideen Shaheed Jawahar, Soumya Swaminathan, Esaki M. Shankar
2016 PLoS ONE  
Contact investigation is an active case finding strategy to increase detection of Tuberculosis (TB) and a key component of TB control programs. The household contacts are at a higher risk of exposure than members of the general population. The information on the value and yield of household contact screening and the approaches used in high incidence settings like India is limited. Objective To evaluate the yield of active case finding in household contacts of newly diagnosed smear positive TB
more » ... tients and the factors associated with increased yield. Method Retrospective record review of the household contacts of newly diagnosed sputum smear positive patients (index case) enrolled in a clinical trial at National Institute of Research in Tuberculosis, Chennai during the period 2007-2014. A sequential screening algorithm with chest x-ray followed by symptom screen was employed to identify presumptive TB patients. Results 643 household contacts of 280 index TB patients were identified out of which 544 (85%) consented for screening. 71/544 (13%) patients had an abnormal chest radiograph and out of them 70% were symptomatic. A total of 29/544 (5.3%) contacts were found to have TB among whom 23/29 (79%) were sputum smear positive. The number needed to screen (NNS) to identify a new TB case among all household contacts was 19 and among those with an abnormal CXR was 02. Age group > 44 years, male gender and siblings of the index PLOS ONE |
doi:10.1371/journal.pone.0162090 pmid:27583974 pmcid:PMC5008766 fatcat:jdfdk475jvg7zfi5oyam267dam

Structural Basis of Ordered Binding of Donor and Acceptor Substrates to the Retaining Glycosyltransferase, α-1,3-Galactosyltransferase

Ester Boix, Yingnan Zhang, G. Jawahar Swaminathan, Keith Brew, K. Ravi Acharya
2002 Journal of Biological Chemistry  
Bovine ␣-1,3-galactosyltransferase (␣3GT) catalyzes the synthesis of the ␣-galactose (␣-Gal) epitope, the target of natural human antibodies. It represents a family of enzymes, including the histo blood group A and B transferases, that catalyze retaining glycosyltransfer reactions of unknown mechanism. An initial study of ␣3GT in a crystal form with limited resolution and considerable disorder suggested the possible formation of a ␤-galactosyl-enzyme covalent intermediate (Gastinel, 2001) EMBO
more » ... . 20, 638 -649). Highly ordered structures are described for complexes of ␣3GT with donor substrate, UDP-galactose, UDPglucose, and two acceptor substrates, lactose and Nacetyllactosamine, at resolutions up to 1.46 Å. Structural and calorimetric binding studies suggest an obligatory ordered binding of donor and acceptor substrates, linked to a donor substrate-induced conformational change, and the direct participation of UDP in acceptor binding. The monosaccharide-UDP bond is cleaved in the structures containing UDP-galactose and UDPglucose, producing non-covalent complexes containing buried ␤-galactose and ␣-glucose. The location of these monosaccharides and molecular modeling suggest that binding of a distorted conformation of UDP-galactose may be important in the catalytic mechanism of ␣3GT.
doi:10.1074/jbc.m202631200 pmid:12011052 fatcat:fxs75w27drfztlnwf2dmghhjfu

DECIPHER: database for the interpretation of phenotype-linked plausibly pathogenic sequence and copy-number variation

Eugene Bragin, Eleni A. Chatzimichali, Caroline F. Wright, Matthew E. Hurles, Helen V. Firth, A. Paul Bevan, G. Jawahar Swaminathan
2013 Nucleic Acids Research  
This article provides an update to the DECIPHER database, an earlier instance of which has been described elsewhere [Swaminathan et al. (2012) DECIPHER: web-based, community resource for clinical interpretation  ... 
doi:10.1093/nar/gkt937 pmid:24150940 pmcid:PMC3965078 fatcat:vup5lxwbcvehtfcng7txdrr65y

Impact of HIV Infection on the Recurrence of Tuberculosis in South India

Sujatha Narayanan, Soumya Swaminathan, Philip Supply, Sivakumar Shanmugam, Gopalan Narendran, Lalitha Hari, Ranjani Ramachandran, Camille Locht, Mohideen Shaheed Jawahar, Paranji Raman Narayanan
2010 Journal of Infectious Diseases  
doi:10.1086/650528 pmid:20121433 fatcat:hntojtsbfbe5bi3lqlynu2weqe

Straightforward and complete deposition of NMR data to the PDBe

Christopher J. Penkett, Glen van Ginkel, Sameer Velankar, Jawahar Swaminathan, Eldon L. Ulrich, Steve Mading, Tim J. Stevens, Rasmus H. Fogh, Aleksandras Gutmanas, Gerard J. Kleywegt, Kim Henrick, Wim F. Vranken
2010 Journal of Biomolecular NMR  
We present a suite of software for the complete and easy deposition of NMR data to the PDB and BMRB. This suite uses the CCPN framework and introduces a freely downloadable, graphical desktop application called CcpNmr Entry Completion Interface (ECI) for the secure editing of experimental information and associated datasets through the lifetime of an NMR project. CCPN projects can be created within the CcpNmr Analysis software or by importing existing NMR data files using the CcpNmr
more » ... ter. After further data entry and checking with the ECI, the project can then be rapidly deposited to the PDBe using AutoDep, or exported as a complete deposition NMR-STAR file. In full CCPN projects created with ECI, it is straightforward to select chemical shift lists, restraint data sets, structural ensembles and all relevant associated experimental collection details, which all are or will become mandatory when depositing to the PDB. Instructions and download information for the ECI are available from the PDBe web site at
doi:10.1007/s10858-010-9439-3 pmid:20680401 pmcid:PMC2950272 fatcat:dwh4dfnbavfvdgsnvgp3pemj34

The Crystal Structure of Human Placenta Growth Factor-1 (PlGF-1), an Angiogenic Protein, at 2.0 Å Resolution

Shalini Iyer, Demetres D. Leonidas, G. Jawahar Swaminathan, Domenico Maglione, Mauro Battisti, Marina Tucci, M. Graziella Persico, K. Ravi Acharya
2000 Journal of Biological Chemistry  
The angiogenic molecule placenta growth factor (PlGF) is a member of the cysteine-knot family of growth factors. In this study, a mature isoform of the human PlGF protein, PlGF-1, was crystallized as a homodimer in the crystallographic asymmetric unit, and its crystal structure was elucidated at 2.0 Å resolution. The overall structure of PlGF-1 is similar to that of vascular endothelial growth factor (VEGF) with which it shares 42% amino acid sequence identity. Based on structural and
more » ... l data, we have mapped several important residues on the PlGF-1 molecule that are involved in recognition of the fms-like tyrosine kinase receptor (Flt-1, also known as VEGFR-1). We propose a model for the association of PlGF-1 and Flt-1 domain 2 with precise shape complementarity, consider the relevance of this assembly for PlGF-1 signal transduction, and provide a structural basis for altered specificity of this molecule.
doi:10.1074/jbc.m008055200 pmid:11069911 fatcat:desqdkshojd33omdtrbu5ccuam

Structure of UDP Complex of UDP-galactose:β-Galactoside-α-1,3-galactosyltransferase at 1.53-Å Resolution Reveals a Conformational Change in the Catalytically Important C Terminus

Ester Boix, G. Jawahar Swaminathan, Yingnan Zhang, Ramanathan Natesh, Keith Brew, K. Ravi Acharya
2001 Journal of Biological Chemistry  
UDP-galactose:␤-galactosyl ␣-1,3-galactosyltransferase (␣3GT) catalyzes the transfer of galactose from UDP-␣-Dgalactose into an ␣-1,3 linkage with ␤-galactosyl groups in glycoconjugates. The enzyme is expressed in many mammalian species but is absent from humans, apes, and old world monkeys as a result of the mutational inactivation of the gene; in humans, a large fraction of natural antibodies are directed against its product, the ␣-galactose epitope. ␣3GT is a member of a family of
more » ... dent retaining glycosyltransferases including the histo-blood group A and B synthases. A crystal structure of the catalytic domain of ␣3GT was recently reported (Gastinel, 2001) EMBO J. 20, 638 -649). However, because of the limited resolution (2.3 Å) and high mobility of the atoms (as indicated by high B-factors) this structure (form I) does not provide a clear depiction of the catalytic site of the enzyme. Here we report a new, highly ordered structure for the catalytic domain of ␣3GT at 1.53-Å resolution (form II). This provides a more accurate picture of the details of the catalytic site that includes a bound UDP molecule and a Mn 2؉ cofactor. Significantly, in the new structure, the C-terminal segment (residues 358 -368) adopts a very different, highly structured conformation and appears to form part of the active site. The properties of an Arg-365 to Lys mutant indicate that this region is important for catalysis, possibly reflecting its role in a donor substrate-induced conformational change.
doi:10.1074/jbc.m108828200 pmid:11592969 fatcat:3rhgshhcsvgpnhlu7lnrjpsny4

Charcot-Leyden Crystal Protein (Galectin-10) Is Not a Dual Function Galectin with Lysophospholipase Activity but Binds a Lysophospholipase Inhibitor in a Novel Structural Fashion

Steven J. Ackerman, Li Liu, Mark A. Kwatia, Michael P. Savage, Demetres D. Leonidas, G. Jawahar Swaminathan, K. Ravi Acharya
2002 Journal of Biological Chemistry  
Charcot-Leyden crystal (CLC) protein, initially reported to possess weak lysophospholipase activity, is still considered to be the eosinophil's lysophospholipase, but it shows no sequence similarities to any known lysophospholipases. In contrast, CLC protein has moderate sequence similarity, conserved genomic organization, and near structural identity to members of the galectin superfamily, and it has been designated galectin-10. To definitively determine whether or not CLC protein is a
more » ... pholipase, we reassessed its enzymatic activity in peripheral blood eosinophils and an eosinophil myelocyte cell line (AML14.3D10). Antibody affinity chromatography was used to fully deplete CLC protein from eosinophil lysates. The CLC-depleted lysates retained their full lysophospholipase activity, and this activity could be blocked by sulfhydryl group-reactive inhibitors, N-ethylmaleimide and p-chloromercuribenzenesulfonate, previously reported to inhibit the eosinophil enzyme. In contrast, the affinity-purified CLC protein lacked significant lysophospholipase activity. X-ray crystallographic structures of CLC protein in complex with the inhibitors showed that p-chloromercuribenzenesulfonate bound CLC protein via disulfide bonds with Cys 29 and with Cys 57 near the carbohydrate recognition domain (CRD), whereas N-ethylmaleimide bound to the galectin-10 CRD via ring stacking interactions with Trp 72 , in a manner highly analogous to mannose binding to this CRD. Antibodies to rat pancreatic lysophospholipase identified a protein in eosinophil and AML14.3D10 cell lysates, comparable in size with human pancreatic lysophospholipase, which co-purifies in small quantities with CLC protein. Ligand blotting of human and murine eosinophil lysates with CLC protein as probe showed that it binds proteins also recognized by antibodies to pancreatic lysophospholipase. Our results definitively show that CLC protein is not one of the eosinophil's lysophospholipases but that it does interact with eosinophil lysophospholipases and known inhibitors of this lipolytic activity.
doi:10.1074/jbc.m200221200 pmid:11834744 fatcat:sdjgdu65xrernbptlbne6m3xcy

Evaluation of metformin in combination with rifampicin containing antituberculosis therapy in patients with new, smear-positive pulmonary tuberculosis (METRIF): study protocol for a randomised clinical trial

Chandrasekaran Padmapriyadarsini, Perumal K Bhavani, Mohan Natrajan, Chinnayan Ponnuraja, Hemanth Kumar, Sivaramakrishnan N Gomathy, Randeep Guleria, Shaheed M Jawahar, Manjula Singh, Tanjore Balganesh, Soumya Swaminathan
2019 BMJ Open  
IntroductionShorter duration of treatment for the management of drug-susceptible pulmonary tuberculosis (TB) would be a significant improvement in the care of patients suffering from the disease. Besides newer drugs and regimens, other modalities like host-directed therapy are also being suggested to reach this goal. This study's objective is to assess the efficacy and safety of metformin-containing anti-TB treatment (ATT) regimen in comparison to the standard 6-month ATT regimen in the
more » ... t of patients with newly diagnosed sputum smear-positive drug-sensitive pulmonary TB.Methods and analysisWe are conducting a multicentric, randomised open-label controlled clinical trial to achieve the study objective. The intervention group will receive isoniazid (H), rifampicin (R), ethambutol (E) and pyrazinamide (Z) along with 1000 mg of daily metformin (Met) for the first 2 months while the control group will receive only HRZE. After 2 months, both the groups will receive HRE daily for 4 months. The primary endpoint is time to sputum culture conversion. Secondary endpoints will include time to detection of Mycobacterium tuberculosis in sputum, pharmacokinetics and pharmacogenomics of study drugs, drug–drug interactions, safety and tolerability of the various combinations and measurement of autophagy and immune responses in the study participants.Ethics and disseminationThe ethics committee of the participating institutes have approved the study. Results from this trial will contribute to evidence towards constructing a shorter, effective and safe regimen for patients with TB. The results will be shared widely with the National Programme managers, policymakers and stakeholders through open access publications, dissemination meetings, conference abstracts and policy briefs. This is expected to provide a new standard of care for drug-sensitive patients with pulmonary TB who will not only reduce the number of clinic visits and lost to follow-up of patients from treatment but also reduce the burden on the healthcare system.Trial registration numberCTRI/2018/01/011176; Pre-results.
doi:10.1136/bmjopen-2018-024363 pmid:30826761 pmcid:PMC6429929 fatcat:va2kgwrq3bc65jguaclzkvoyb4

Predictors of unfavorable responses to therapy in rifampicin-sensitive pulmonary tuberculosis using an integrated approach of radiological presentation and sputum mycobacterial burden

Narendran Gopalan, Vignes Anand Srinivasalu, Ponnuraja Chinnayan, Banurekha Velayutham, Adhin Bhaskar, Ramesh Santhanakrishnan, Thirumaran Senguttuvan, Sridhar Rathinam, Mahilmaran Ayyamperumal, Kumar Satagopan, Dhanalakshmi Rajendran, Tamizhselvan Manoharan (+16 others)
2021 PLoS ONE  
Introduction Despite the exalted status of sputum mycobacterial load for gauging pulmonary tuberculosis treatment and progress, Chest X-rays supplement valuable information for taking instantaneous therapeutic decisions, especially during the COVID-19 pandemic. Even though literature on individual parameters is overwhelming, few studies have explored the interaction between radiographic parameters denoting severity with mycobacterial burden signifying infectivity. By using a sophisticated
more » ... ch of integrating Chest X-ray parameters with sputum mycobacterial characteristics, evaluated at all the three crucial time points of TB treatment namely pre-treatment, end of intensive phase and completion of treatment, utilizing the interactive Cox Proportional Hazards model, we aimed to precisely deduce predictors of unfavorable response to TB treatment. Materials and method We extracted de-identified data from well characterized clinical trial cohorts that recruited rifampicin-sensitive Pulmonary TB patients without any comorbidities, taking their first spell of anti-tuberculosis therapy under supervision and meticulous follow up for 24 months post treatment completion, to accurately predict TB outcomes. Radiographic data independently obtained, interpreted by two experienced pulmonologists was collated with demographic details and, sputum smear and culture grades of participants by an independent statistician and analyzed using the Cox Proportional Hazards model, to not only adjust for confounding factors including treatment effect, but also explore the interaction between radiological and bacteriological parameters for better therapeutic application. Results Of 667 TB patients with data available, cavitation, extent of involvement, lower zone involvement, smear and culture grade at baseline were significant parameters predisposing to an unfavorable TB treatment outcome in the univariate analysis. Reduction in radiological lesions in Chest X-ray by at least 50% at 2 months and 75% at the end of treatment helped in averting unfavorable responses. Smear and Culture conversion at the end of 2 months was highly significant as a predictor (p<0.001). In the multivariate analysis, the adjusted hazards ratios (HR) for an unfavorable response to TB therapy for extent of involvement, baseline cavitation and persistence (post treatment) were 1.21 (95% CI: 1.01–1.44), 1.73 (95% CI: 1.05–2.84) and 2.68 (95% CI: 1.4–5.12) respectively. A 3+ smear had an HR of 1.94 (95% CI: 0.81–4.64). Further probing into the interaction, among patients with 3+ and 2+ smears, HRs for cavitation were 3.26 (95% CI: 1.33–8.00) and 1.92 (95% CI: 0.80–4.60) while for >2 zones, were 3.05 (95% CI: 1.12–8.23) and 1.92 (95% CI: 0.72–5.08) respectively. Patients without cavitation, zonal involvement <2, and a smear grade less than 2+ had a better prognosis and constituted minimal disease. Conclusion Baseline Cavitation, Opacities occupying >2 zones and 3+ smear grade individually and independently forecasted a poorer TB outcome. The interaction model revealed that Zonal involvement confined to 2 zones, without a cavity and smear grade up to 2+, constituting "minimal disease", had a better prognosis. Radiological clearance >50% along with smear conversion at the end of intensive phase of treatment, observed to be a reasonable alternative to culture conversion in predicting a successful outcome. These parameters may potentially take up key positions as stratification factors for future trials contemplating on shorter TB regimens.
doi:10.1371/journal.pone.0257647 pmid:34543329 pmcid:PMC8452066 fatcat:vwnj2itklbd65e3urqpjndl4n4

Genetic diagnosis of developmental disorders in the DDD study: a scalable analysis of genome-wide research data

Caroline F Wright, Tomas W Fitzgerald, Wendy D Jones, Stephen Clayton, Jeremy F McRae, Margriet van Kogelenberg, Daniel A King, Kirsty Ambridge, Daniel M Barrett, Tanya Bayzetinova, A Paul Bevan, Eugene Bragin (+20 others)
2015 The Lancet  
Human genome sequencing has transformed our understanding of genomic variation and its relevance to health and disease, and is now starting to enter clinical practice for the diagnosis of rare diseases. The question of whether and how some categories of genomic fi ndings should be shared with individual research participants is currently a topic of international debate, and development of robust analytical workfl ows to identify and communicate clinically relevant variants is paramount. Methods
more » ... The Deciphering Developmental Disorders (DDD) study has developed a UK-wide patient recruitment network involving over 180 clinicians across all 24 regional genetics services, and has performed genome-wide microarray and whole exome sequencing on children with undiagnosed developmental disorders and their parents. After data analysis, pertinent genomic variants were returned to individual research participants via their local clinical genetics team. Findings Around 80 000 genomic variants were identifi ed from exome sequencing and microarray analysis in each individual, of which on average 400 were rare and predicted to be protein altering. By focusing only on de novo and segregating variants in known developmental disorder genes, we achieved a diagnostic yield of 27% among 1133 previously investigated yet undiagnosed children with developmental disorders, whilst minimising incidental fi ndings. In families with developmentally normal parents, whole exome sequencing of the child and both parents resulted in a 10-fold reduction in the number of potential causal variants that needed clinical evaluation compared to sequencing only the child. Most diagnostic variants identifi ed in known genes were novel and not present in current databases of known disease variation. Interpretation Implementation of a robust translational genomics workfl ow is achievable within a large-scale rare disease research study to allow feedback of potentially diagnostic fi ndings to clinicians and research participants. Systematic recording of relevant clinical data, curation of a gene-phenotype knowledge base, and development of clinical decision support software are needed in addition to automated exclusion of almost all variants, which is crucial for scalable prioritisation and review of possible diagnostic variants. However, the resource requirements of development and maintenance of a clinical reporting system within a research setting are substantial.
doi:10.1016/s0140-6736(14)61705-0 pmid:25529582 pmcid:PMC4392068 fatcat:o6cn3i5eabgyralymswrmlrcye

Rare Variants in NR2F2 Cause Congenital Heart Defects in Humans

Saeed Al Turki, Ashok K. Manickaraj, Catherine L. Mercer, Sebastian S. Gerety, Marc-Phillip Hitz, Sarah Lindsay, Lisa C.A. D'Alessandro, G. Jawahar Swaminathan, Jamie Bentham, Anne-Karin Arndt, Jacoba Low, Jeroen Breckpot (+27 others)
2014 American Journal of Human Genetics  
doi:10.1016/j.ajhg.2014.06.013 pmcid:PMC4085635 fatcat:5it2pk7u7nenlhcnm6vkggt6oa
« Previous Showing results 1 — 15 out of 123 results