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  1. 3:58 pm

Tuesday, June 5

  1. page Carbamate Kinase, Giardia Lamblia edited ... */Disease Information (sort of like the Intro to your Mini Research Write up): Giardiasis is …
    ...
    */Disease Information (sort of like the Intro to your Mini Research Write up):
    Giardiasis is a common intestinal infection caused by the exposure to the organism Giardia lamblia [1]. Exposure to contaminated water, food, or direct contact with the organism can lead to the disease [1]. Giardia is one of the most common intestinal parasites in the world making Giardiasis a very common infection. Cases are becoming increasingly common in the United States. A well-known cause of Giardiasis is through drinking unfiltered water that may be contaminated with human or animal waste [1]. This has brought the infectious disease to be highly associated with nicknames “travelers diarrhea” and “beaver fever”. One of the main concerns is that children can easily spread the infection in a daycare environment. The increased contact between children and each other’s feces makes it a likely spreading ground [1]. When the inactive forms of Giardia (Cysts) are ingested, the acid within the stomach releases the parasite from the cyst. From there the parasite latches to the lining of the small intestine and reproduces. Cysts are then passed on through the fecal matter of the host [1]. Giardiasis is non fatal and a large number of those infected never display symptoms. When symptoms are displayed they often involve watery and aggressive diarrhea other symptoms include stomach pain, nausea, and vomiting. The disease can often be allowed to run its course or treated with a multitude of antibiotics.
    ...
    ed.), 1889-1892
    Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.)
    https://www.ncbi.nlm.nih.gov/gene/5701142
    ...
    ATP + NH3 + CO2
    -- link to Sigma (or other company) page for assay (see Sigma links below)
    https://www.sigmaaldrich.com/catalog/product/supelco/46856u?lang=en&region=US
    https://www.sigmaaldrich.com/catalog/product/sigma/flaas?lang=en&region=US
    https://www.sigmaaldrich.com/catalog/product/supelco/46856u?lang=en®ion=US
    https://www.sigmaaldrich.com/catalog/product/sigma/flaas?lang=en®ion=US

    Structure (PDB or Homology model)
    -- PDB # or closest PDB entry if using homology model: 4OLC
    ...
    Molar Extinction coefficient of your protein at 280 nm wavelength: Ext. coefficient 28420 Abs 0.1% (=1 g/l) 0.830, assuming all Cys residues are reduced
    TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
    {https://embnet.vital-it.ch/download.php?file=TMPRED.10313.6576.gif}{https://embnet.vital-it.ch/download.php?file=TMPRED.2283.801.gif} results
    *CDS Gene Sequence (paste as text only):
    GGACCTGCTAACCCTTGCTGTGGTGTCTACAGGAGTCACTGCGGGGGCGCTTTCCTCCATAACACTTGTA
    (view changes)
    12:18 pm

Sunday, May 13

  1. page Lysyl tRNA Synthetase edited <span style="color: #000000; font-family: Arial,sans-serif;">Gene Sequence: <…

    <span style="color: #000000; font-family: Arial,sans-serif;">Gene Sequence: </span><span style="color: #000000;">>NC_008611.1:c4654901-4653405 Mycobacterium ulcerans Agy99, complete genome</span>
    GTGAGCTCCGCCGAGTCAAACATTCCCGAGCAGTTCCGAATCCGTCGGGACAAGCGTGCTCGGTTATTGG
    CAGAGGGGTATGACCCCTATCCGGTAGCCATCGAACGCACTCACACGTTGGCCGAGATTCGTGCCACCTA
    TGCCGACCTACCGACAGATTCGGCGACCGAGGACATCGTCGGTGTCGCGGGTCGAGTGGTCTTCGCGCGT
    AACACCGGGAAACTGTGTTTTGCGACACTGCAGGACGGCGACGGCACCCAGCTGCAGGCGATGATCAGCC
    TCGACGAGGTGGGCCGCGAATCACTGGACAGGTGGAAGGCCGACGTCGACATCGGTGACGTCGTCTACGT
    GCACGGCACCGTGATCAGCTCCCGTCGGGGCGAACTATCGGTGCTCGCCGATTCCTGGCGGATGGCTGCC
    AAGGCCTTGCGGCCGCTGCCGGTCGCACACAAGGAGATGAGCGAGGAGTCGCGGGTTCGTCAGCGCTACG
    TCGATTTGATCGTGCGTCCACAGGCGCGGGAGGTCGCGCGACAGCGGATTGCGGTAATCCGTGCGGTGCG
    TAATGCGCTGGAGCGGCGCGGGTTTCTCGAAGTCGAAACGCCAATGTTGCAGACCTTGGCCGGCGGCGCT
    GCGGCGCGGCCGTTCGTCACCCATTCCAATGCCCTCGACATCGATCTTTACCTGCGTATCGCGCCGGAAC
    TGTTCCTCAAGCGTTGCATCGTGGGCGGTTTCGACAGGGTCTTTGAACTAAATCGGGTGTTCCGAAACGA
    AGGGTCTGATTCCACCCATTCCCCGGAATTCTCCATGCTGGAGACGTATCAGACCTATGGAACCTATGAC
    GATTCGGCCCTGATCACCAGAGAGCTTATTCAAGAAGTTGCTGACGAGGCGATCGGAACCAGACAACTCT
    CGATGCCTGACGGCAGCGTGTACGACATCGACGGAGAATGGGCAACTATGGAGATGTATTCCTCCTTGTC
    GGAGGCACTGGGGGAACAGATCACGCCAGAGACGACGGTTGCTCGCTTACGTGACATCGCAAGCGGACTT
    GACGTAGAGATTGACAACAGTGTTTTTGGTCACGGCAAACTTGTCGAGGAACTCTGGGAACATGCTGTCG
    GCAACAAGTTGACGGCGCCCACATTTGTCAAGGATTTCCCAGTCGAGACAACGCCATTGACTCGTCAGCA
    CCGCAGTATCCCCGGAGTAACCGAGAAGTGGGATCTCTACGTGCGCGGAGTCGAACTGGCTACCGGGTAC
    TCGGAATTAAACGATCCCGTCGTCCAGCGGGACAGATTCGCTGATCAGGCCCGTGCGGCGGCCGCGGGTG
    ACGACGAGGCGATGCAACTTGACGAAGATTTTCTGACGGCTCTGGAGTACGGGATGCCCCCGTGTACTGG
    AACTGGAATGGGCATCGATCGGTTGTTGATGTGTTTGACCGGACTGTCAATTAGGGAGACAGTTTTGTTC
    CCGATTGTTCGCCCTCACTCCAACTGA
    BRENDA Link:
    https://www.brenda-enzymes.org/enzyme.php?ecno=6.1.1.6
    *Target (protein/gene name): Lysyl-tRNA Synthetase
    [[code]]
    <span style="font-size: 10pt;">*NCBI Gene # or RefSeq#:NC_008611.1</span>
    <span style="font-size: 10pt;">*Protein ID (NP or XP #) or Wolbachia#:ABL06216</span>
    <span style="font-size: 10pt;">*Organism (including strain)://Mcobacterium ulcerans// (Agy99)</span>
    <span style="font-size: 10pt;">Etiologic Risk Group (see link below): Unknown</span>
    <span style="font-size: 10pt;">*/Disease Information (sort of like the Intro to your Mini Research Write up):Buruli Ulcer is caused by the protozoan //Mycobacterium ulcerans// in which the flesh is destroyed in patches of the body. Transmission method is currently unknown as it is found primarily in neglected populations. It is present in every habitable continent, but is only highly prevalent in underdeveloped tropical regions.</span>
    <span style="font-size: 10pt;">Link to TDR Targets page (if present):None</span>
    <span style="font-size: 10pt;">Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.)</span>
    <span style="font-size: 10pt;">Essentiality of this protein: Essential in other mycobaterium</span>
    <span style="font-size: 10pt;"> Is it a monomer or multimer as biological unit? (make prediction at </span><span style="font-size: 10pt;">[[@http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html]]</span><span style="font-size: 10pt;">):</span>
    <span style="font-size: 10pt;">Complex of proteins?:Single protein</span>
    <span style="font-size: 10pt;">Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):</span><span style="font-size: 10pt;">[[https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5091859/]]</span>
    <span style="font-size: 10pt;">Lysyl tRNA synthetase is druggable in worm parasites by affecting the single protein.</span>
    [[code]]
    *EC#:
    [[code]]
    <span style="font-size: 10pt;">Link to BRENDA EC# page: </span>
    <span style="font-size: 10pt;">[[https://www.brenda-enzymes.org/enzyme.php?ecno=6.1.1.6]]</span>
    <span style="font-size: 10pt;">[[https://www.brenda-enzymes.org/pathway_index.php?menu=off&site=pathway&search_mode=exact&pathway=lysine_metabolism&ecno=6.1.1.6]]</span>
    <span style="font-size: 10pt;">[[image:https://vdsstream.wikispaces.com/site/embedthumbnail/placeholder?w=624&h=351 width="624" height="351"]]</span>
    <span style="font-size: 10pt;">Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):</span>
    <span style="font-size: 10pt;">-- link to Sigma (or other company) page for assay (see Sigma links below)</span>
    <span style="font-size: 10pt;">-- -or link (or citation) to paper that contains assay https://www.sciencedirect.com/science/article/pii/030090849290158B?via%3Dihubinformationhttps://www.sciencedirect.com/science/article/pii/030090849290158B?via%3Dihub</span>
    <span style="font-size: 10pt;">-- links to assay reagents (substrates) pages.http://www.bio-rad.com/en-us/sku/5000006-bio-rad-protein-assay-dye-reagent-concentrate-450-ml?ID=5000006</span>
    <span style="font-size: 10pt;">--- List cost and quantity of substrate reagents, supplier, and catalog #$129.00 for Bio-Rad Protein Assay Reagent, 450 ml #5000006</span>
    [[code]]
    Structure (PDB or Homology model)
    [[code]]
    <span style="font-size: 10pt;"> -- PDB # or closest PDB entry if using homology model:</span>
    <span style="font-size: 10pt;"> -- For Homology Model option:</span>
    <span style="font-size: 10pt;"> ---- Show pairwise alignment of your BLASTP search in NCBI against the PDB</span>
    <span style="font-size: 10pt;"> ---- Query Coverage:</span>
    <span style="font-size: 10pt;"> ---- Max % Identities:</span>
    <span style="font-size: 10pt;"> ---- % Positives</span>
    <span style="font-size: 10pt;"> ---- Chain used for homology:</span>
    [[code]]
    Current Inhibitors: None for mycobacterium
    [[code]]
    <span style="font-size: 10pt;">Expression Information (has it been expressed in bacterial cells):E. coli BL21 (DE3)</span>
    <span style="font-size: 10pt;">Purification Method:https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1432-1033.1978.tb12064.x</span>
    <span style="font-size: 10pt;">Image of protein (PyMol with features delineated and shown separately): [[image:https://vdsstream.wikispaces.com/site/embedthumbnail/placeholder?w=624&h=468 width="624" height="468"]]</span>
    <span style="font-size: 10pt;">*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): MAHHHHHHMSSAESNIPEQFRIRRDKRARLLAEGYDPYPVAIERTHTLAEIRATYADLPTDSATEDIVGVAGRVVFARNT</span>
    <span style="font-size: 10pt;">GKLCFATLQDGDGTQLQAMISLDEVGRESLDRWKADVDIGDVVYVHGTVISSRRGELSVLADSWRMAAKALRPLPVAHKE</span>
    <span style="font-size: 10pt;">MSEESRVRQRYVDLIVRPQAREVARQRIAVIRAVRNALERRGFLEVETPMLQTLAGGAAARPFVTHSNALDIDLYLRIAP</span>
    <span style="font-size: 10pt;">ELFLKRCIVGGFDRVFELNRVFRNEGSDSTHSPEFSMLETYQTYGTYDDSALITRELIQEVADEAIGTRQLSMPDGSVYD</span>
    <span style="font-size: 10pt;">IDGEWATMEMYSSLSEALGEQITPETTVARLRDIASGLDVEIDNSVFGHGKLVEELWEHAVGNKLTAPTFVKDFPVETTP</span>
    <span style="font-size: 10pt;">LTRQHRSIPGVTEKWDLYVRGVELATGYSELNDPVVQRDRFADQARAAAAGDDEAMQLDEDFLTALEYGMPPCTGTGMGI</span>
    <span style="font-size: 10pt;">DRLLMCLTGLSIRETVLFPIVRPHSN</span>
    <span style="font-size: 10pt;">*length of your protein in Amino Acids</span>
    <span style="font-size: 10pt;">506</span>
    <span style="font-size: 10pt;">Molecular Weight of your protein in kiloDaltons using the [[http://expasy.org/tools/protparam.html|Expasy ProtParam]] website</span>
    56511.80
    [[code]]
    Molar Extinction coefficient of your protein at 280 nm wavelength:<span style="color: #626262;"> 48360</span>
    TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it. {https://vdsstream.wikispaces.com/site/embedthumbnail/placeholder?w=624&h=468} results
    [[code]]
    <span style="font-size: 10pt;">*CDS Gene Sequence (paste as text only):>lcl|ORF1</span>
    <span style="font-size: 10pt;">MISLDEVGRESLDRWKADVDIGDVVYVHGTVISSRRGELSVLADSWRMAA</span>
    <span style="font-size: 10pt;">KALRPLPVAHKEMSEESRVRQRYVDLIVRPQAREVARQRIAVIRAVRNAL</span>
    <span style="font-size: 10pt;">ERRGFLEVETPMLQTLAGGAAARPFVTHSNALDIDLYLRIAPELFLKRCI</span>
    <span style="font-size: 10pt;">VGGFDRVFELNRVFRNEGSDSTHSPEFSMLETYQTYGTYDDSALITRELI</span>
    <span style="font-size: 10pt;">QEVADEAIGTRQLSMPDGSVYDIDGEWATMEMYSSLSEALGEQITPETTV</span>
    <span style="font-size: 10pt;">ARLRDIASGLDVEIDNSVFGHGKLVEELWEHAVGNKLTAPTFVKDFPVET</span>
    <span style="font-size: 10pt;">TPLTRQHRSIPGVTEKWDLYVRGVELATGYSELNDPVVQRDRFADQARAA</span>
    <span style="font-size: 10pt;">AAGDDEAMQLDEDFLTALEYGMPPCTGTGMGIDRLLMCLTGLSIRETVLF</span>
    <span style="font-size: 10pt;">PIVRPHSN</span>
    <span style="font-size: 10pt;">*GC% Content for gene:58.717435</span>
    <span style="font-size: 10pt;">*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):>lcl|ORF1</span>
    <span style="font-size: 10pt;">MISLDEVGRESLDRWKADVDIGDVVYVHGTVISSRRGELSVLADSWRMAA</span>
    <span style="font-size: 10pt;">KALRPLPVAHKEMSEESRVRQRYVDLIVRPQAREVARQRIAVIRAVRNAL</span>
    <span style="font-size: 10pt;">ERRGFLEVETPMLQTLAGGAAARPFVTHSNALDIDLYLRIAPELFLKRCI</span>
    <span style="font-size: 10pt;">VGGFDRVFELNRVFRNEGSDSTHSPEFSMLETYQTYGTYDDSALITRELI</span>
    <span style="font-size: 10pt;">QEVADEAIGTRQLSMPDGSVYDIDGEWATMEMYSSLSEALGEQITPETTV</span>
    <span style="font-size: 10pt;">ARLRDIASGLDVEIDNSVFGHGKLVEELWEHAVGNKLTAPTFVKDFPVET</span>
    <span style="font-size: 10pt;">TPLTRQHRSIPGVTEKWDLYVRGVELATGYSELNDPVVQRDRFADQARAA</span>
    <span style="font-size: 10pt;">AAGDDEAMQLDEDFLTALEYGMPPCTGTGMGIDRLLMCLTGLSIRETVLF</span>
    <span style="font-size: 10pt;">PIVRPHSN</span>
    <span style="font-size: 10pt;">*GC% Content for gene (codon optimized):58.717435</span>
    [[code]]
    Do Not Need this info for Spring (but still copy these lines to your Target page for now)
    [[code]]
    <span style="font-size: 10pt;">Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):</span>
    <span style="font-size: 10pt;">(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)</span>
    <span style="font-size: 10pt;"> -- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.</span>
    [[code]]
    Primer design results for 'tail' primers (this is just 2 sequences):

    (view changes)
    12:24 pm
  2. page Targets edited ... TargetSp18 - Lactate dehydogenase (//Cryptosporidium parvum//)................................…
    ...
    TargetSp18 - Lactate dehydogenase (//Cryptosporidium parvum//)..............................................................................................................................................Jacky X.
    TargetSp18 - Dengue Methyltransferase........................................................................................................................................................................................Tom S.
    TargetSp18- Lysyl tRNA Synthetase (Mycobacterium ulcerans).............................................................................................................Tyler G.
    Summer 2017 Target Discovery
    Revivals
    (view changes)
    12:23 pm

Monday, May 7

  1. page Dihydrofolate Reductase (Bacillus anthracis) edited ... Etiological Risk Group: Category B Priority Pathogens -Bacillus anthracis Disease: ... [2,…
    ...
    Etiological Risk Group: Category B Priority Pathogens -Bacillus anthracis
    Disease:
    ...
    [2, 3].
    http://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0004689#abstract0
    Homology: Bacillus cereus, Bacillus thuringiensis, and Bacillus mycoides
    ...
    For Homology Model option: N/A
    Expression System: Escherichia coli BL21(DE3)
    PairwiseInhibitors: n/a
    Pairwise
    alignment of
    ...
    against the PDB--99%PDB--100% aligned
    {Capture (2).PNG}
    https://blast.ncbi.nlm.nih.gov/Blast.cgi#alnHdr_446669140
    Gene Availability:
    http://www.uniprot.org/proteomes/UP000000427
    https://www.ncbi.nlm.nih.gov/gene/3361714

    Protein sequence:
    >2QK8:A|PDBID|CHAIN|SEQUENCE
    ...
    EEVFELCKNEEEIFIFGGAQIYDLFLPYVDKLYITKIHHAFEGDTFFPEMDMTNWKEVFVEKGLTDEKNPYTYYYHVYEK
    QQ
    length of your protein in Amino Acids: 162 aa
    Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website: 19.124 kDa
    Molar Extinction coefficient of your protein at 280 nm wavelength: 1.439
    TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html).
    {TMPRED.1664.2779.gif}
    CDS Gene Sequence:
    <span style="background-color: #ffffff; font-family: monospace,serif;">>NC_001496.1:c135455-133161 Bacillus anthracis virulence plasmid PX01, complete sequence
    ATGAAAAAACGAAAAGTGTTAATACCATTAATGGCATTGTCTACGATATTAGTTTCAAGCACAGGTAATT
    TAGAGGTGATTCAGGCAGAAGTTAAACAGGAGAACCGGTTATTAAATGAATCAGAATCAAGTTCCCAGGG
    GTTACTAGGATACTATTTTAGTGATTTGAATTTTCAAGCACCCATGGTGGTTACCTCTTCTACTACAGGG
    GATTTATCTATTCCTAGTTCTGAGTTAGAAAATATTCCATCGGAAAACCAATATTTTCAATCTGCTATTT
    GGTCAGGATTTATCAAAGTTAAGAAGAGTGATGAATATACATTTGCTACTTCCGCTGATAATCATGTAAC
    AATGTGGGTAGATGACCAAGAAGTGATTAATAAAGCTTCTAATTCTAACAAAATCAGATTAGAAAAAGGA
    AGATTATATCAAATAAAAATTCAATATCAACGAGAAAATCCTACTGAAAAAGGATTGGATTTCAAGTTGT
    ACTGGACCGATTCTCAAAATAAAAAAGAAGTGATTTCTAGTGATAACTTACAATTGCCAGAATTAAAACA
    AAAATCTTCGAACTCAAGAAAAAAGCGAAGTACAAGTGCTGGACCTACGGTTCCAGACCGTGACAATGAT
    GGAATCCCTGATTCATTAGAGGTAGAAGGATATACGGTTGATGTCAAAAATAAAAGAACTTTTCTTTCAC
    CATGGATTTCTAATATTCATGAAAAGAAAGGATTAACCAAATATAAATCATCTCCTGAAAAATGGAGCAC
    GGCTTCTGATCCGTACAGTGATTTCGAAAAGGTTACAGGACGGATTGATAAGAATGTATCACCAGAGGCA
    AGACACCCCCTTGTGGCAGCTTATCCGATTGTACATGTAGATATGGAGAATATTATTCTCTCAAAAAATG
    AGGATCAATCCACACAGAATACTGATAGTCAAACGAGAACAATAAGTAAAAATACTTCTACAAGTAGGAC
    ACATACTAGTGAAGTACATGGAAATGCAGAAGTGCATGCGTCGTTCTTTGATATTGGTGGGAGTGTATCT
    GCAGGATTTAGTAATTCGAATTCAAGTACGGTCGCAATTGATCATTCACTATCTCTAGCAGGGGAAAGAA
    CTTGGGCTGAAACAATGGGTTTAAATACCGCTGATACAGCAAGATTAAATGCCAATATTAGATATGTAAA
    TACTGGGACGGCTCCAATCTACAACGTGTTACCAACGACTTCGTTAGTGTTAGGAAAAAATCAAACACTC
    GCGACAATTAAAGCTAAGGAAAACCAATTAAGTCAAATACTTGCACCTAATAATTATTATCCTTCTAAAA
    ACTTGGCGCCAATCGCATTAAATGCACAAGACGATTTCAGTTCTACTCCAATTACAATGAATTACAATCA
    ATTTCTTGAGTTAGAAAAAACGAAACAATTAAGATTAGATACGGATCAAGTATATGGGAATATAGCAACA
    TACAATTTTGAAAATGGAAGAGTGAGGGTGGATACAGGCTCGAACTGGAGTGAAGTGTTACCGCAAATTC
    AAGAAACAACTGCACGTATCATTTTTAATGGAAAAGATTTAAATCTGGTAGAAAGGCGGATAGCGGCGGT
    TAATCCTAGTGATCCATTAGAAACGACTAAACCGGATATGACATTAAAAGAAGCCCTTAAAATAGCATTT
    GGATTTAACGAACCGAATGGAAACTTACAATATCAAGGGAAAGACATAACCGAATTTGATTTTAATTTCG
    ATCAACAAACATCTCAAAATATCAAGAATCAGTTAGCGGAATTAAACGCAACTAACATATATACTGTATT
    AGATAAAATCAAATTAAATGCAAAAATGAATATTTTAATAAGAGATAAACGTTTTCATTATGATAGAAAT
    AACATAGCAGTTGGGGCGGATGAGTCAGTAGTTAAGGAGGCTCATAGAGAAGTAATTAATTCGTCAACAG
    AGGGATTATTGTTAAATATTGATAAGGATATAAGAAAAATATTATCAGGTTATATTGTAGAAATTGAAGA
    TACTGAAGGGCTTAAAGAAGTTATAAATGACAGATATGATATGTTGAATATTTCTAGTTTACGGCAAGAT
    GGAAAAACATTTATAGATTTTAAAAAATATAATGATAAATTACCGTTATATATAAGTAATCCCAATTATA
    AGGTAAATGTATATGCTGTTACTAAAGAAAACACTATTATTAATCCTAGTGAGAATGGGGATACTAGTAC
    CAACGGGATCAAGAAAATTTTAATCTTTTCTAAAAAAGGCTATGAGATAGGATAA
    </span>

    (view changes)
    12:18 pm
  2. file Capture (2).PNG uploaded
    12:14 pm
  3. file Capture.PNG uploaded
    12:12 pm
  4. 11:31 am

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