Clostridium thermocellum is a well-characterized cellulose-degrading microorganism. The genome sequence of C. thermocellum encodes a number of proteins that contain type I dockerin domains, which ...implies that they are components of the cellulose-degrading apparatus, but display no significant sequence similarity to known plant cell wall-degrading enzymes. Here, we report the biochemical properties and crystal structure of one of these proteins, designated CtCel124. The protein was shown to be an endo-acting cellulase that displays a single displacement mechanism and acts in synergy with Cel48S, the major cellulosomal exo-cellulase. The crystal structure of CtCel124 in complex with two cellotriose molecules, determined to 1.5 à , displays a superhelical fold in which a constellation of α-helices encircle a central helix that houses the catalytic apparatus. The catalytic acid, Glu96, is located at the C-terminus of the central helix, but there is no candidate catalytic base. The substrate-binding cleft can be divided into two discrete topographical domains in which the bound cellotriose molecules display twisted and linear conformations, respectively, suggesting that the enzyme may target the interface between crystalline and disordered regions of cellulose.
Abstract This longitudinal study investigated the 3-D facial soft-tissue response to transverse palatal bone-anchored osteodistraction in 18 adult patients. Laser-scanned facial surface data were ...obtained for all patients before (T0 ), 6 months (T1 ) and 1 year (T2 ) after transverse palatal distraction. The averaged facial morphologies at T0 , T1 and T2 were calculated and compared. Sagittal and vertical measurements were obtained from lateral cephalograms to evaluate skeletal and dental movements. Pre- and immediate post-distraction dental casts were used to investigate transverse maxillary movements. Cutaneous changes were mainly observed in the paranasal regions and cheeks, in the range 1–3 mm, reflecting the underlying increase in the maxillary width. A significant enlargement of the nasal base was also demonstrated. The absolute magnitude of these facial changes was limited but clinically relevant. Variable skeletal movements were observed. These were of low magnitude and no systematic tendency could be demonstrated statistically. Significant changes were documented only with regard to orthodontic repositioning of the upper and lower incisors. The mean transverse increases in the maxillary dental arch were 6.7 mm at the intercanine, 6.8 mm at the interpremolar, and 6.1 mm at the intermolar levels.
Resistance to osimertinib in advanced EGFR-mutated non-small cell lung cancer (NSCLC) constitutes a significant challenge for clinicians either in terms of molecular diagnosis and subsequent ...therapeutic implications.
This is a prospective single-centre study with the primary objective of characterising resistance mechanisms to osimertinib in advanced EGFR-mutated NSCLC patients treated both in first- and in second-line. Next-Generation Sequencing analysis was conducted on paired tissue biopsies and plasma samples. A concordance analysis between tissue and plasma was performed.
Sixty-five advanced EGFR-mutated NSCLC patients treated with osimertinib in first- (n = 56) or in second-line (n = 9) were included. We managed to perform tissue and liquid biopsies in 65.5% and 89.7% of patients who experienced osimertinib progression, respectively. Acquired resistance mechanisms were identified in 80% of 25 patients with post-progression samples, with MET amplification (n = 8), EGFR C797S (n = 3), and SCLC transformation (n = 2) the most frequently identified. The mean concordance rates between tissue and plasma for the EGFR activating mutation and for the molecular resistance mechanisms were 87.5% and 22.7%, respectively.
Resistance to osimertinib demonstrated to be highly heterogeneous, with MET amplification the main mechanism. Plasma genotyping is a relevant complementary tool which might integrate tissue analysis for the study of resistance mechanisms.
Monoclonal antibodies (ICI) targeting the immune checkpoint PD-1/PD-L1 alone or in combination with chemotherapy have demonstrated relevant benefits and established new standards of care in ...first-line treatment for advanced non-oncogene addicted non-small cell lung cancer (NSCLC). However, a relevant percentage of NSCLC patients, even with high PD-L1 expression, did not respond to ICI, highlighting the presence of intracellular resistance mechanisms that could be dependent on high PD-L1 levels. The intracellular signaling induced by PD-L1 in tumor cells and their correlation with angiogenic signaling pathways are not yet fully elucidated.
The intrinsic role of PD-L1 was initially checked in two PD-L1 overexpressing NSCLC cells by transcriptome profile and kinase array. The correlation of PD-L1 with VEGF, PECAM-1, and angiogenesis was evaluated in a cohort of advanced NSCLC patients. The secreted cytokines involved in tumor angiogenesis were assessed by Luminex assay and their effect on Huvec migration by a non-contact co-culture system.
PD-L1 overexpressing cells modulated pathways involved in tumor inflammation and JAK-STAT signaling. In NSCLC patients, PD-L1 expression was correlated with high tumor intra-vasculature. When challenged with PBMC, PD-L1 overexpressing cells produced higher levels of pro-angiogenic factors compared to parental cells, as a consequence of STAT signaling activation. This increased production of cytokines involved in tumor angiogenesis largely stimulated Huvec migration. Finally, the addition of the anti-antiangiogenic agent nintedanib significantly reduced the spread of Huvec cells when exposed to high levels of pro-angiogenic factors.
In this study, we reported that high PD-L1 modulates STAT signaling in the presence of PBMC and induces pro-angiogenic factor secretion. This could enforce the role of PD-L1 as a crucial regulator of the tumor microenvironment stimulating tumor progression, both as an inhibitor of T-cell activity and as a promoter of tumor angiogenesis.
Gunther von Hagens' development of plastination as a method for preserving human remains has enabled his public display of skinless, dissected bodies in a series of popular international exhibitions ...entitled Body Worlds. These spectacular displays claim to be educative, democratizing the study of anatomy and liberating it from the traditional confines of professional medical study. However, Body Worlds has raised various ethical objections to its commercial purpose, sourcing of some bodies and arrangement of bodies in poses or dissections that some viewers find offensive. Here we consider a different, often overlooked ethical conundrum raised by these exhibitions: the likelihood that the viewing of plastinates posed in 'frozen motion' is ill‑suited to the psychological development of young children (5-10 years old) whose understanding of death is still in formation. Often young children mistake corpses for models, even for living beings if they are posed in arrested motion. The educative value of Body Worlds for younger viewers is questionable and the display may even interfere with their understanding of death. If the exhibition of human remains can be justified where their authenticity can be made known to viewers and the remains invested by them with sympathetic emotional meaning, it may be pointless if not unethical to show quasi‑lifelike posed plastinates to young children in lieu of replica models.
The aim of this study was to evaluate the effects of scanning parameters on the precision of the data acquired using a facial laser scanner and to assess the reliability of automatic model recording ...in humans.
Data were acquired using a laser scanner (Cyberware 3030RGB); analysis and measurements were performed with Rapid Form 2004 software. A mannequin and six volunteers were scanned to investigate the effects of environmental conditions, positioning, head orientation, and software procedures. Precision and accuracy of the data were evaluated comparing six linear measures calculated on scanned data with those obtained directly. Two sessions with different head inclination were performed. The reliability of repeated scans was also assessed measuring the distance between the surfaces reconstructed from two separate scans of the same subject, at 12 anatomical points, in 5 subjects, during two sessions using a different head inclination. Differences were analysed using paired t-tests or analysis of variance (ANOVA).
The accuracy of scanning was +/-0.65 mm. The development of a specific protocol resulted in a mean scanning error of 1-1.2 mm and a recording error of 0.3-0.4 mm on repeated scans of human subjects.
This study indicates that scanning of the human face may be hampered by errors and artefacts, mainly due to movements. While the effect of trembling and involuntary movements during the exam may be minimized using faster scanning devices, comparative observation over time may be affected by unreal differences due to the uncertainty of facial expression. The overall error is, however, in the range useful for most clinical studies.
Purpose
To prospectively evaluate the efficacy and safety of RIRS, SWL and PCNL for lower calyceal stones sized 1–2 cm.
Materials and methods
Patients with a single lower calyceal stone with an ...evidence of a CT diameter between 1 and 2 cm were enrolled in this multicenter, randomized, unblinded, clinical trial study. Patients were randomized into three groups: group A: SWL (194 pts); group B: RIRS (207 pts); group C: PCNL (181 pts). Patients were evaluated with KUB radiography (US for uric acid stones) at day 10 and a CT scan after 3 months. The CONSORT 2010 statement was adhered to where possible. The collected data were analyzed.
Results
The mean stone size was 13.78 mm in group A, 14.82 mm in group B and 15.23 mm in group C (
p
= 0.34). Group C compared to group B showed longer operative time 72.3 vs. 55.8 min (
p
= 0.082), fluoroscopic time 175.6 vs. 31.8 min (
p
= 0.004) and hospital stay 3.7 vs. 1.3 days (
p
= 0.039). The overall stone-free rate (SFR) was 61.8% for group A, 82.1% for group B and 87.3% for group C. The re-treatment rate was significantly higher in group A compared to the other two groups, 61.3% (
p
< 0.05). The auxiliary procedure rate was comparable for groups A and B and lower for group C (
p
< 0.05). The complication rate was 6.7, 14.5 and 19.3% for groups A, B and C, respectively.
Conclusions
RIRS and PCNL were more effective than SWL to obtain a better SFR and less auxiliary and re-treatment rate in single lower calyceal stone with a CT diameter between 1 and 2 cm. RIRS compared to PCNL offers the best outcome in terms of procedure length, radiation exposure and hospital stay.
ISRCTN 55546280.
Abstract Objectives To compare simple enucleation (SE) and standard partial nephrectomy (SPN) in terms of surgical results in a multicenter dataset (RECORd Project). Materials and methods patients ...treated with nephron sparing surgery (NSS) for clinical T1 renal tumors between January 2009 and January 2011 were evaluated. Overall, 198 patients who underwent SE were retrospectively matched to 198 patients who underwent SPN. The SPN and SE groups were compared regarding intraoperative, early post-operative and pathologic outcome variables. Multivariable analysis was applied to analyze predictors of positive surgical margin (PSM) status. Results SE was associated with similar WIT (18 vs 17.8 min), lower intraoperative blood loss (177 vs 221 cc, p = 0.02) and shorter operative time (121 vs 147 min; p < 0.0001). Surgical approach (laparoscopic vs. open), tumor size and type of indication (elective/relative vs absolute) were associated with WIT >20 min. The incidence of PSM was significantly lower in patients treated with SE (1.4% vs 6.9%; p = 0.02). At multivariable analysis, PSM was related to the surgical technique, with a 4.7-fold increased risk of PSM for SPN compared to SE. The incidence of overall, medical and surgical complications was similar between SE and SPN. Conclusions Type of NSS technique (SE vs SPN) adopted has a negligible impact on WIT and postoperative morbidity but SE seems protective against PSM occurrence.
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