Journal: Annals of Gastroenterological Surgery

Article Title: Tele‐proctoring for minimally invasive surgery across Japan: An initial step toward a new approach to improving the disparity of surgical care and supporting surgical education

doi: 10.1002/ags3.12750

Figure Lengend Snippet: Implemented tele‐proctoring with Sapporo Medical University between January 2021 and September 2022.

Article Snippet: Sapporo Medical University , Keio University , 1150 km , FTTB , 100 , 24 → 3.0 , 10.0 , 1.0 , 24.5 (23.0–27.0).

Techniques:

Journal: Nature microbiology

Article Title: FttA is a CPSF73 homologue that terminates transcription in Archaea

doi: 10.1038/s41564-020-0667-3

Figure Lengend Snippet: a. and b. TECs +125 (lane 3) resume elongation upon NTP addition to generate +225 nt transcripts −/+ 25 mM DPA (lanes 4-11). FttA addition results in transcript cleavage and release of most TECs to the supernatant - DPA (lanes 12-13), inhibiting resumed elongation upon NTP addition (lanes 14-15). Pre-incubation of FttA with 25 mM DPA inhibits FttA-mediated termination (lanes 16-17), permitting TECs +125 to resume elongation (lanes 18-19). Lanes M1 and M2 contain 32 P-labeled 10- and 50-nt ssDNA markers, respectively. Similar results were observed in 5 independent experiments. c. Inhibition of metallo-beta-lactamase/beta-CASP protein activity impairs growth of T. kodakarensis. A mid-log culture of T. kodakarensis strain TS559 was split into nine cultures, with three biological replicates exposed to 0 mM (peach), 12.5 mM (green) or 25 mM DPA (purple). d, e and f . RNAs recovered one-hour post DPA-addition to cultures of TS559, or from cultures of IR5 grown in the absence of NaF display altered 3'-termini. TRIzol extracted RNAs were reverse transcribed with primers complementary to nascent transcript sequences of TK1146, TK0222 and TK0676 to generate cDNAs that were quantified and normalized to internal controls. Inhibiting FttA-activity with DPA or lowering steady-state FttA levels by riboswitch-mediated controlled expression impacts the abundance of RNAs with extended 3’-termini in vivo . RNA abundance in untreated TS559 cultures (open bars) was set to 1.0, and fold changes in the abundance of amplicons reflecting RNA transcripts with extended 3’-sequences at increasing distances from the translation stop site (purple, green, orange and red) are shown for strain IR5 (solid bars), TS559 + 12.5 mM DPA (wide stripes) and TS559 + 25 mM DPA (narrow stripes). Errors were calculated at a 95% confidence interval with the center value as the mean of 3 biological replicates. g. Maps of the TK1428 locus in parental (TS559), N-terminally tagged (TK1428D) and riboswitch-regulated expression (IR5) strains. h. Western blots demonstrate the reduction in FttA protein levels in strain IR5 upon removal of NaF from the medium; n = 3 independent replicates. Size standards are identified by MW (left).

Article Snippet: Purified, recombinant full-length FttA was used as an antigen to prepare polyclonal antibodies in mice (Cocalico Biologicals).

Techniques: Incubation, Labeling, Inhibition, Activity Assay, Reverse Transcription, Expressing, In Vivo, Western Blot

Journal: Nature microbiology

Article Title: FttA is a CPSF73 homologue that terminates transcription in Archaea

doi: 10.1038/s41564-020-0667-3

Figure Lengend Snippet: a . Transcripts within intact TECs are retained in pellet (P) fractions; transcripts released from terminated TECs partition into the supernatant (S). Radiolabeled transcripts within starting material (SM) TECs +125 and mock treated TECs +125 are retained in pellet fractions (lanes 1-4), whereas FttA WT addition results in cleavage of nascent transcripts and termination of most TECs (lanes 5-6). A catalytically deficient FttA variant (FttA H255A ) abrogates cleavage and RNA release (lanes 7-8). Lane M contains 32 P-labeled ssDNA markers. b. FttA-mediated termination is distinct from RNase treatment of intact TECs. TECs +125 (SM, lane 1) are resistant to repeated washes and readily resume elongation upon NTP addition to generate +225 nt transcripts (lanes 2-5). Dashed boxes and arrows denote +125 transcripts that are elongated to +225 nt transcripts; the specific activity of +225 transcripts can be increased by addition of additional 32 P-α-UTP during resumed elongation. RNase I f digestion of nascent transcripts associated with washed TECs +125 results in degradation of the nascent transcript to just ~20-30 nts, but TECs with shortened transcripts remain associated with the DNA and survive repeated washing (lanes 10-11). TECs ~+25-30 resultant from RNase I f treatment of TECs +125 readily resume elongation upon NTP addition to generate ~+125 nt full-length transcripts (lanes 12-13). Dashed boxes and arrows denote ~+25 transcripts that are elongated to ~+125 nt transcripts; the specific activity of ~+125 transcripts can be increased by addition of 32 P-α-UTP during resumed elongation. FttA addition to TECs+125 disrupts most TECs with nascent transcript cleavage (lanes 6-9), results in release of most TECs from the template and cleaved transcripts cannot be extended by NTP addition (lanes 8-9). c and d . Diagrams of the fate of TECs +125 following RNase I f and FttA treatment, respectively. e. FttA releases RNAP from the DNA template into solution confirming dissociation of the TEC and bona fide FttA-mediated transcription termination. RNAP is tracked and quantified by Western blots (n = 3 independent replicates) with anti-HA antibodies that recognize the modified RpoL-subunit. f. FttA is not reliant on NTP hydrolysis to inactivate TECs, cleave nascent transcripts and terminate transcription. For panels a, b f: Similar results were observed in 4 independent experiments.

Article Snippet: Purified, recombinant full-length FttA was used as an antigen to prepare polyclonal antibodies in mice (Cocalico Biologicals).

Techniques: Variant Assay, Labeling, Activity Assay, Western Blot, Modification

Journal: Nature microbiology

Article Title: FttA is a CPSF73 homologue that terminates transcription in Archaea

doi: 10.1038/s41564-020-0667-3

Figure Lengend Snippet: a. Promoter-directed transcription of biotinylated templates encoding G-less or C-less cassettes permits formation of TECs with increasing length A-, C-, and U-rich, or A-, G-, and U-rich nascent transcripts, respectively. FL = full-length; all templates permit elongation for 100 nts beyond the G- or C-less cassette. b. TECs remain stably associated and transcripts are primarily recovered in the pellet (P) fraction in the absence (−) of FttA. When FttA is present (+), transcripts are cleaved and primarily recovered in the supernatant (S) fraction. Cleavage releases ~20 – 30 nt shorter transcripts (boxed). The left-most lane contains 32 P-labeled ssDNA markers. c. Addition of Spt4-Spt5 largely abrogates the RNA sequence-requirements of FttA-mediated transcription termination. T= total reaction = P+S. The left-most lane contains 32 P-labeled ssDNA markers. d. Transcript release was quantified with and without FttA addition for TECs with increasing length transcripts on G-less (pink/salmon) and C-less cassettes (mint/green), with and without Spt4-Spt5 addition for TECs +125 formed on G- and C-less cassettes. Error bars were calculated as standard deviation from the mean (n = 3 independent experiments). For panels b, c: Similar results were observed in 3 independent experiments.

Article Snippet: Purified, recombinant full-length FttA was used as an antigen to prepare polyclonal antibodies in mice (Cocalico Biologicals).

Techniques: Stable Transfection, Labeling, Sequencing, Standard Deviation

Journal: Nature microbiology

Article Title: FttA is a CPSF73 homologue that terminates transcription in Archaea

doi: 10.1038/s41564-020-0667-3

Figure Lengend Snippet: a and b . Washed, NTP-deprived TECs +125 were assembled on biotinylated templates with a +125 nt G-less cassette. Resumed elongation upon differential [NTP] addition permits transcription to generate +225 nt transcripts, albeit at different rates. c . FttA readily terminates stalled or slowly elongating TECs (lanes 17-24) and FttA-mediated termination becomes competitive with transcription elongation even at high [NTP] in the presence of Spt4-Spt5 (lanes 25-32). Similar results were observed in 3 independent experiments.

Article Snippet: Purified, recombinant full-length FttA was used as an antigen to prepare polyclonal antibodies in mice (Cocalico Biologicals).

Techniques:

Journal: Radiology

Article Title: PET of Poly (ADP-Ribose) Polymerase Activity in Cancer: Preclinical Assessment and First In-Human Studies

doi: 10.1148/radiol.2016161929

Figure Lengend Snippet: Synthesis scheme and structure of 18F-FTT fluorthanatrace. DMSO = dimethyl sulfoxide, OMs = mesylate, OTs = tosylate.

Article Snippet: Whole-body images (midskull to midcalf) were obtained after injecting a mean (±standard deviation) of 374 MBq ± 19 (range, 348–403 MBq [0.5–2.1 µg]) of 18 F-FTT fluorthanatrace with a PET/computed tomographic (CT) scanner (Biograph 40; Siemens, Knoxville, Tenn) by using 50 mAs (effective) for the attenuation-correction CT scan.

Techniques:

Journal: Radiology

Article Title: PET of Poly (ADP-Ribose) Polymerase Activity in Cancer: Preclinical Assessment and First In-Human Studies

doi: 10.1148/radiol.2016161929

Figure Lengend Snippet: 18F-FTT fluorthanatrace micro-PET images of athymic nude mice demonstrate nodal and spine activity before and after treatment with olaparib or iniparib. Coronal and sagittal micro-PET (last 10 minutes of 60-minute acquisition) and CT images of normal mice with lymph nodes and spine uptake indicated by yellow arrowheads or white region of interest. Corresponding time-activity curves are also shown. Left and right lymph node standard uptake values (SUV) over time were averaged to create graphs of time-activity curves for lymph nodes.

Article Snippet: Whole-body images (midskull to midcalf) were obtained after injecting a mean (±standard deviation) of 374 MBq ± 19 (range, 348–403 MBq [0.5–2.1 µg]) of 18 F-FTT fluorthanatrace with a PET/computed tomographic (CT) scanner (Biograph 40; Siemens, Knoxville, Tenn) by using 50 mAs (effective) for the attenuation-correction CT scan.

Techniques: Micro-PET, Activity Assay

Journal: Radiology

Article Title: PET of Poly (ADP-Ribose) Polymerase Activity in Cancer: Preclinical Assessment and First In-Human Studies

doi: 10.1148/radiol.2016161929

Figure Lengend Snippet: Boxplots of effect of olaparib and iniparib on 18F-FTT fluorthanatrace uptake (expressed as AUC area under the receiver operating characteristic curve) in axillary lymph nodes and marrow of spine. * indicates P = .0001 when compared with pretreatment value.

Article Snippet: Whole-body images (midskull to midcalf) were obtained after injecting a mean (±standard deviation) of 374 MBq ± 19 (range, 348–403 MBq [0.5–2.1 µg]) of 18 F-FTT fluorthanatrace with a PET/computed tomographic (CT) scanner (Biograph 40; Siemens, Knoxville, Tenn) by using 50 mAs (effective) for the attenuation-correction CT scan.

Techniques:

Journal: Radiology

Article Title: PET of Poly (ADP-Ribose) Polymerase Activity in Cancer: Preclinical Assessment and First In-Human Studies

doi: 10.1148/radiol.2016161929

Figure Lengend Snippet: Representative maximum intensity projection18F-FTT fluorthanatrace PET images from three different healthy volunteers demonstrate normal spleen, node, and marrow uptake. All images are scaled to maximum standardized uptake value (SUV) of 10.

Article Snippet: Whole-body images (midskull to midcalf) were obtained after injecting a mean (±standard deviation) of 374 MBq ± 19 (range, 348–403 MBq [0.5–2.1 µg]) of 18 F-FTT fluorthanatrace with a PET/computed tomographic (CT) scanner (Biograph 40; Siemens, Knoxville, Tenn) by using 50 mAs (effective) for the attenuation-correction CT scan.

Techniques:

Journal: Radiology

Article Title: PET of Poly (ADP-Ribose) Polymerase Activity in Cancer: Preclinical Assessment and First In-Human Studies

doi: 10.1148/radiol.2016161929

Figure Lengend Snippet: 18F-FTT fluorthanatrace PET (left), unenhanced CT (middle), and fused transaxial (right) images in 52-year-old man with recurrent pancreatic adenocarcinoma after Whipple procedure (subject 2, Table 2). Images were obtained 30 minutes after tracer injection. Arrows indicate location of recurrent pancreatic cancer. Gray-scale PET images were set to same scale as that shown for color image. SUV = standardized uptake value.

Article Snippet: Whole-body images (midskull to midcalf) were obtained after injecting a mean (±standard deviation) of 374 MBq ± 19 (range, 348–403 MBq [0.5–2.1 µg]) of 18 F-FTT fluorthanatrace with a PET/computed tomographic (CT) scanner (Biograph 40; Siemens, Knoxville, Tenn) by using 50 mAs (effective) for the attenuation-correction CT scan.

Techniques: Injection

Journal: Radiology

Article Title: PET of Poly (ADP-Ribose) Polymerase Activity in Cancer: Preclinical Assessment and First In-Human Studies

doi: 10.1148/radiol.2016161929

Figure Lengend Snippet: From left to right, transaxial 18F-FTT fluorthanatrace PET image, unenhanced CT scan, fused image, and contrast material–enhanced MR image in a 62-year-old woman with metastatic biphenotypic hepatocellular carcinoma/cholangiocarcinoma (subject 8, Table 2). The subject had previously undergone right hepatic trisegmentectomy and treatment with multiple chemotherapy regimens. The most recent chemotherapy regimen is described in Table 2. MR imaging was performed 1 month before 18F-FTT fluorthanatrace PET/CT and demonstrated new metastatic liver lesions with decreased signal intensity, as seen on MR image (arrow), in addition to enlargement of left hepatic lobe as a result of previous partial hepatic resection. CT (liver window) showed interval enlargement of metastasis, as demonstrated by low-attenuation lesion that is clearly larger than low-signal-intensity lesion seen on corresponding MR image. 18F-FTT fluorthanatrace image shows increased uptake within this metastasis. PET/CT images were obtained 150 minutes after tracer injection. MR image was obtained 20 minutes after gadoxetate disodium injection. Arrows show locations of metastatic lesions. Gray-scale PET images were set to same scale as that shown for color image. SUV = standardized uptake value.

Article Snippet: Whole-body images (midskull to midcalf) were obtained after injecting a mean (±standard deviation) of 374 MBq ± 19 (range, 348–403 MBq [0.5–2.1 µg]) of 18 F-FTT fluorthanatrace with a PET/computed tomographic (CT) scanner (Biograph 40; Siemens, Knoxville, Tenn) by using 50 mAs (effective) for the attenuation-correction CT scan.

Techniques: Computed Tomography, Imaging, Positron Emission Tomography-Computed Tomography, Injection

Journal: Radiology

Article Title: PET of Poly (ADP-Ribose) Polymerase Activity in Cancer: Preclinical Assessment and First In-Human Studies

doi: 10.1148/radiol.2016161929

Figure Lengend Snippet: Transaxial 18F-FTT fluorthanatrace PET (left), unenhanced CT (middle), and fused (right) images in 67-year-old man with recurrent pancreatic adenocarcinoma (subject 6, Table 2). 18F-FTT fluorthanatrace PET/CT was performed 18 days after completing radiation treatment (3000 cGy in 10 fractions over 2 weeks). Images were obtained 90 minutes after tracer injection. Arrows indicate tumor location. Green lines were used to aid in tumor localization given the lack of contrast on CT images and lack of uptake on 18F-FTT fluorthanatrace PET images. Gray-scale PET images were set to same scale as that shown for color image.

Article Snippet: Whole-body images (midskull to midcalf) were obtained after injecting a mean (±standard deviation) of 374 MBq ± 19 (range, 348–403 MBq [0.5–2.1 µg]) of 18 F-FTT fluorthanatrace with a PET/computed tomographic (CT) scanner (Biograph 40; Siemens, Knoxville, Tenn) by using 50 mAs (effective) for the attenuation-correction CT scan.

Techniques: Positron Emission Tomography-Computed Tomography, Injection