Management & surveillance of rectal neuroendocrine tumours: a single-centre retrospective analysis.

BACKGROUND: Rectal neuroendocrine tumours (rNETs) are rare but are increasing in incidence. Current management and surveillance recommendations are based on low-grade evidence. Follow-up practices are often inconsistent and costly. This retrospective study analyses a single-centre's experience with rNETs to assess incidence, management practices, outcomes, and guideline adherence. METHODS: This is a single-centre retrospective study from Queensland Australia, spanning from 2012 to 2023. Twenty-eight rNET cases met inclusion criteria. Examined parameters included incidence, management, outcomes and adherence to European Neuroendocrine Tumour Society (ENETS) guidelines. R1 resection rate was analysed for associations with resection technique and lesion recognition and recurrence rate was assessed in all patients. RESULTS: This study shows an increasing incidence of rNETs during the study period, reflecting a global trend. R1 resection rate at initial endoscopy was 75%. There was a general lack of advanced endoscopic techniques utilized and poor lesion recognition, however a statistically significant correlation was not established between these factors and an R1 result (P < 0.05). Most patients with an R1 result had subsequent re-resection to render the result R0, however five patients (33%) underwent surveillance with no reports of recurrence on follow-up. Overall, follow-up practices in our cohort were inconsistent and did not adhere to guidelines. CONCLUSION: rNETs are increasing in incidence, emphasizing the need for standardized management and surveillance. Further training is required for rNET recognition and advanced endoscopic resection techniques. Further research is required to assess long-term outcomes in surveilled R1 cases, understand optimal endoscopic resection techniques and further develop local surveillance guidelines.

R-Loop Accumulation in Spliceosome Mutant Leukemias Confers Sensitivity to PARP1 Inhibition by Triggering Transcription-Replication Conflicts.

RNA splicing factor (SF) gene mutations are commonly observed in patients with myeloid malignancies. Here we showed that SRSF2- and U2AF1-mutant leukemias are preferentially sensitive to PARP inhibitors (PARPi), despite being proficient in homologous recombination repair. Instead, SF-mutant leukemias exhibited R-loop accumulation that elicited an R-loop-associated PARP1 response, rendering cells dependent on PARP1 activity for survival. Consequently, PARPi induced DNA damage and cell death in SF-mutant leukemias in an R-loop-dependent manner. PARPi further increased aberrant R-loop levels, causing higher transcription-replication collisions and triggering ATR activation in SF-mutant leukemias. Ultimately, PARPi-induced DNA damage and cell death in SF-mutant leukemias could be enhanced by ATR inhibition. Finally, the level of PARP1 activity at R-loops correlated with PARPi sensitivity, suggesting that R-loop-associated PARP1 activity could be predictive of PARPi sensitivity in patients harboring SF gene mutations. This study highlights the potential of targeting different R-loop response pathways caused by spliceosome gene mutations as a therapeutic strategy for treating cancer. SIGNIFICANCE: Spliceosome-mutant leukemias accumulate R-loops and require PARP1 to resolve transcription-replication conflicts and genomic instability, providing rationale to repurpose FDA-approved PARP inhibitors for patients carrying spliceosome gene mutations.

SNMMI Clinical Trials Network Research Series for Technologists: Imaging Contract Research Organizations, Nuclear Medicine Technologists, and the Role They Play in Medical Imaging Research.

Clinical imaging research is a fast-growing, complex, and integral part of drug and therapy discovery and development. Research sponsors rely on outside vendors to manage their trials and deliver results they hope will demonstrate the efficacy of their product. Specialized vendors known as imaging contract research organizations have teams of highly trained and specialized professionals who lend their expertise to all aspects of imaging research management, of which nuclear medicine technologists are key team members. This article is part of the Clinical Trials Network Research Series for Technologists and will help provide an overview of an imaging research study from initiation to data delivery and the roles that nuclear medicine technologists and other imaging professionals play.

Heterologous production of the D-cycloserine intermediate O-acetyl-L-serine in a human type II pulmonary cell model.

Tuberculosis (TB) is the second leading cause of death by a single infectious disease behind COVID-19. Despite a century of effort, the current TB vaccine does not effectively prevent pulmonary TB, promote herd immunity, or prevent transmission. Therefore, alternative approaches are needed. We seek to develop a cell therapy that produces an effective antibiotic in response to TB infection. D-cycloserine (D-CS) is a second-line antibiotic for TB that inhibits bacterial cell wall synthesis. We have determined D-CS to be the optimal candidate for anti-TB cell therapy due to its effectiveness against TB, relatively short biosynthetic pathway, and its low-resistance incidence. The first committed step towards D-CS synthesis is catalyzed by the L-serine-O-acetyltransferase (DcsE) which converts L-serine and acetyl-CoA to O-acetyl-L-serine (L-OAS). To test if the D-CS pathway could be an effective prophylaxis for TB, we endeavored to express functional DcsE in A549 cells as a human pulmonary model. We observed DcsE-FLAG-GFP expression using fluorescence microscopy. DcsE purified from A549 cells catalyzed the synthesis of L-OAS as observed by HPLC-MS. Therefore, human cells synthesize functional DcsE capable of converting L-serine and acetyl-CoA to L-OAS demonstrating the first step towards D-CS production in human cells.

Inpatient Mortality and 30-Day Readmission Rates Associated with Troponin Testing in Patients without Acute Myocardial Infarction.

OBJECTIVE: Troponin values above the threshold established to diagnose acute myocardial infarction (AMI; >99th percentile) are commonly detected in patients with diagnoses other than AMI. The objective of this study was to compare inpatient mortality and 30-day readmission rate in patients with troponin I (TnI) above and below the 99th percentile in those with type 1 AMI and type 2 myocardial injury. METHODS: Between January 1, 2016 and December 31, 2016, there were 56,895 inpatient hospitalizations; of these 14,326 (25.2%) patients received troponin testing. We evaluated mortality and readmissions in the entire cohort based on the primary discharge International Classification of Diseases, Tenth Edition (ICD-10) diagnosis and grouped into type 1 AMI versus other diagnoses comprising the type 2 AMI group (including ICD-10 codes for congestive heart failure, sepsis, and other). Among those with TnI drawn, we evaluated in-hospital mortality and 30-day readmissions based on troponin values > 99th percentile (≥ 0.1 ng/ml). RESULTS: Among the entire cohort, the inpatient mortality rate was significantly higher in those with TnI testing (5.0%, 95% CI 4.6%-5.3%) compared to those without testing (0.7%, 95% CI 0.6%-0.7%, P < 0.01). In the tested cohort 3,743 (26%) patients had troponin levels above the 99th percentile (> 0.1 ng/ml), and 10,583 (74%) had troponin levels below the 99th percentile (≤ 0.1 ng/ml). Comparing type 2 AMI with type 1 AMI and troponin testing, TnI values ≥ 0.1 ng/ml were associated with higher inpatient mortality (11.6% vs. 3.9%) and 30-day readmission rates (16.9% vs. 10.7%). CONCLUSIONS: A higher inpatient mortality and 30-day readmission rates were found in patients with type 2 AMI compared to type 1 AMI group.

Frequency induced rotation of high-contrast angular intensity fringes from an uncoated SPP device.

High-contrast angular intensity fringes are generated by reflecting laser light from an uncoated spiral phase plate (SPP) device for the first time, to the best of our knowledge. As the laser frequency going into the device is tuned, the fringes rotate. Measured transverse fringe patterns match their theoretical predicted values. They have unity contrast, and their measured intensity varies with laser frequency in a fashion similar to a Fabry-Perot etalon. This effect can be used to enable new miniature devices for angle metrology, imaging, and microscopy.

Fragmentation and inefficiencies in US equity markets: Evidence from the Dow 30.

Using the most comprehensive source of commercially available data on the US National Market System, we analyze all quotes and trades associated with Dow 30 stocks in calendar year 2016 from the vantage point of a single and fixed frame of reference. We find that inefficiencies created in part by the fragmentation of the equity marketplace are relatively common and persist for longer than what physical constraints may suggest. Information feeds reported different prices for the same equity more than 120 million times, with almost 64 million dislocation segments featuring meaningfully longer duration and higher magnitude. During this period, roughly 22% of all trades occurred while the SIP and aggregated direct feeds were dislocated. The current market configuration resulted in a realized opportunity cost totaling over $160 million, a conservative estimate that does not take into account intra-day offsetting events.

Fostering Diversity and Inclusion in the National Heart, Lung, and Blood Institute's Small Business Program.

The National Heart, Lung, and Blood Institute has developed a strategic approach to fostering diversity and inclusion within its community of small business innovators. The approach is focused on three central goals: 1) Increase awareness of NHLBI's support for small businesses among underrepresented groups through focused outreach; 2) Identify and eliminate barriers to entry into small business funding programs and entrepreneurial activities for minority and female applicants through specific, targeted training and support; and 3) Expand diversity and inclusivity within our networks by providing additional support for NHLBI-funded small businesses to hire members of underrepresented groups. Key partnerships with biomedical accelerators in underserved regions will be leveraged to maximize impact and achieve all three goals. This article describes the historical context and current state of policies in this arena; it also provides details about mechanisms and approaches used to achieve the goals.

Growth and Development at the Sphenoethmoidal Junction in Perinatal Primates.

Integration of the sphenoid and ethmoid bones during early postnatal development is poorly described in the literature. A uniquely prolonged patency of sphenoethmoidal synchondrosis or prespheno-septal synchondrosis (PSept) has been attributed to humans. However, the sphenoethmoidal junction has not been studied using a comparative primate sample. Here, we examined development of the sphenoethmoidal interface using ontogenetic samples of Old and New World monkeys, strepsirrhine primates (lemurs and lorises), and a comparative sample of other mammals. Specimens ranging from late fetal to 1 month postnatal age were studied using histology, immunohistochemistry, and micro-computed tomography methods. Our results demonstrate that humans are not unique in anterior cranial base growth at PSept, as it is patent in all newborn primates. We found two distinctions within our sample. First, nearly all primates exhibit an earlier breakdown of the nasal capsule cartilage that abuts the orbitosphenoid when compared to nonprimates. This may facilitate earlier postnatal integration of the basicranium and midface and may enhance morphological plasticity in the region. Second, the PSept exhibits a basic dichotomy between strepsirrhines and monkeys. In strepsirrhines, the PSept has proliferating chondrocytes that are primarily oriented in a longitudinal plane, as in other mammals. In contrast, monkeys have a convex anterior end of the presphenoid with a radial boundary of cartilaginous growth at PSept. Our findings suggest that the PSept acts as a "pacemaker" of longitudinal facial growth in mammals with relatively long snouts, but may also contribute to facial height and produce a relatively taller midface in anthropoid primates. Anat Rec, 300:2115-2137, 2017. © 2017 Wiley Periodicals, Inc.

Human language reveals a universal positivity bias.

Using human evaluation of 100,000 words spread across 24 corpora in 10 languages diverse in origin and culture, we present evidence of a deep imprint of human sociality in language, observing that (i) the words of natural human language possess a universal positivity bias, (ii) the estimated emotional content of words is consistent between languages under translation, and (iii) this positivity bias is strongly independent of frequency of word use. Alongside these general regularities, we describe interlanguage variations in the emotional spectrum of languages that allow us to rank corpora. We also show how our word evaluations can be used to construct physical-like instruments for both real-time and offline measurement of the emotional content of large-scale texts.

Electromyographic analysis of muscle activation during sit-and-reach flexibility tests.

The sit-and-reach test (SRT) has been included in standard fitness test batteries for decades, but empirical evidence of actual muscle activity has been lacking. Furthermore, the positioning of the ankle joint during the execution of this test has received relatively scant attention. Therefore, the purpose of this investigation was to compare surface electromyographic (sEMG) activity of selected lower extremity and back musculature and examine the impact of ankle positioning during the standard SRT and the modified sit-and-reach test (MSRT). Seven male and 7 female subjects performed 3 trials of the SRT and MSRT, each in a dorsiflexed and plantar flexed ankle position. During all trials, muscle activity (sEMG) was measured from the right semimembranous (SM), erector spinae (ES), and gastrocnemius (G). Mean sEMG data from each muscle (SM, ES, and G) were normalized by being expressed as a percent contribution to the total electrical activity (100%). Surface electromyographic activity data were also used to determine muscle activation ratios (e.g., SM to ES). Results revealed significantly higher flexibility scores during the plantar flexion condition for both test modalities. The SM exhibited the greatest percent contribution to total sEMG activity within all testing conditions. The SM to G and SM to ES muscle activation ratios were significantly greater than their inverse counterparts within all 4 testing conditions. Based on the 2 sEMG analysis techniques, the SM seemed to exhibit the greatest muscle activity. This investigation provides direct evidence of sEMG muscle activity during the SRT and MSRT, further confirming these tests to be a valid measure of hamstring flexibility.

Functional screening and in vitro analysis reveal thioesterases with enhanced substrate specificity profiles that improve short-chain fatty acid production in Escherichia coli.

Short-chain fatty acid (SCFA) biosynthesis is pertinent to production of biofuels, industrial compounds, and pharmaceuticals from renewable resources. To expand on Escherichia coli SCFA products, we previously implemented a coenzyme A (CoA)-dependent pathway that condenses acetyl-CoA to a diverse group of short-chain fatty acyl-CoAs. To increase product titers and reduce premature pathway termination products, we conducted in vivo and in vitro analyses to understand and improve the specificity of the acyl-CoA thioesterase enzyme, which releases fatty acids from CoA. A total of 62 putative bacterial thioesterases, including 23 from the cow rumen microbiome, were inserted into a pathway that condenses acetyl-CoA to an acyl-CoA molecule derived from exogenously provided propionic or isobutyric acid. Functional screening revealed thioesterases that increase production of saturated (valerate), unsaturated (trans-2-pentenoate), and branched (4-methylvalerate) SCFAs compared to overexpression of E. coli thioesterase tesB or native expression of endogenous thioesterases. To determine if altered thioesterase acyl-CoA substrate specificity caused the increase in product titers, six of the most promising enzymes were analyzed in vitro. Biochemical assays revealed that the most productive thioesterases rely on promiscuous activity but have greater specificity for product-associated acyl-CoAs than for precursor acyl-CoAs. In this study, we introduce novel thioesterases with improved specificity for saturated, branched, and unsaturated short-chain acyl-CoAs, thereby expanding the diversity of potential fatty acid products while increasing titers of current products. The growing uncertainty associated with protein database annotations denotes this study as a model for isolating functional biochemical pathway enzymes in situations where experimental evidence of enzyme function is absent.

The Detection of Motion by Blind Subjects With the Epiretinal 60-Electrode (Argus II) Retinal Prosthesis.

OBJECTIVE: To investigate the ability of 28 blind subjects implanted with a 60-electrode Argus II (Second Sight Medical Products Inc) retinal prosthesis system to detect the direction of a moving object. METHODS: Blind subjects (bare light perception or worse in both eyes) with retinitis pigmentosa were implanted with the Argus II prosthesis as part of a phase 1/2 feasibility study at multiple clinical sites worldwide. The experiment measured their ability to detect the direction of motion of a high-contrast moving bar on a flatscreen monitor in 3 conditions: with the prosthesis system on and a 1-to-1 mapping of spatial information, with the system off, and with the system on but with randomly scrambled spatial information. RESULTS: Fifteen subjects (54%) were able to perform the task significantly better with their prosthesis system than they were with their residual vision, 2 subjects had significantly better performance with their residual vision, and no difference was found for 11 subjects. Of the 15 better-performing subjects, 11 were available for follow-up testing, and 10 of them had significantly better performance with normal rather than with scrambled spatial information. CONCLUSIONS: This work demonstrates that blind subjects implanted with the Argus II retinal prosthesis were able to perform a motion detection task they could not do with their native vision, confirming that electrical stimulation of the retina provides spatial information from synchronized activation of multiple electrodes. TRIAL REGISTRATION: clinicaltrials.gov Identifier:NCT00407602

Analyses of MbtB, MbtE, and MbtF suggest revisions to the mycobactin biosynthesis pathway in Mycobacterium tuberculosis.

The production of mycobactin (MBT) by Mycobacterium tuberculosis is essential for this bacterium to access iron when it is in an infected host. Due to this essential function, there is considerable interest in deciphering the mechanism of MBT assembly, with the goal of targeting select biosynthetic steps for antituberculosis drug development. The proposed scheme for MBT biosynthesis involves assembly of the MBT backbone by a hybrid nonribosomal peptide synthetase (NRPS)/polyketide synthase (PKS) megasynthase followed by the tailoring of this backbone by N(6) acylation of the central l-Lys residue and subsequent N(6)-hydroxylation of the central N(6)-acyl-l-Lys and the terminal caprolactam. A complete testing of this hypothesis has been hindered by the inability to heterologously produce soluble megasynthase components. Here we show that soluble forms of the NRPS components MbtB, MbtE, and MbtF are obtained when these enzymes are coproduced with MbtH. Using these soluble enzymes we determined the amino acid specificity of each adenylation (A) domain. These results suggest that the proposed tailoring enzymes are actually involved in precursor biosynthesis since the A domains of MbtE and MbtF are specific for N(6)-acyl-N(6)-hydroxy-l-Lys and N(6)-hydroxy-l-Lys, respectively. Furthermore, the preference of the A domain of MbtB for l-Thr over l-Ser suggests that the megasynthase produces MBT derivatives with ß-methyl oxazoline rings. Since the most prominent form of MBT produced by M. tuberculosis lacks this ß-methyl group, a mechanism for demethylation remains to be discovered. These results suggest revisions to the MBT biosynthesis pathway while also identifying new targets for antituberculosis drug development.

Metagenomic analysis of Streptomyces lividans reveals host-dependent functional expression.

Most functional metagenomic studies have been limited by the poor expression of many genes derived from metagenomic DNA in Escherichia coli, which has been the predominant surrogate host to date. To expand the range of expressed genes, we developed tools for construction and functional screening of metagenomic libraries in Streptomyces lividans. We expanded on previously published protocols by constructing a system that enables retrieval and characterization of the metagenomic DNA from biologically active clones. To test the functionality of these methods, we constructed and screened two metagenomic libraries in S. lividans. One was constructed with pooled DNA from 14 bacterial isolates cultured from Alaskan soil and the second with DNA directly extracted from the same soil. Functional screening of these libraries identified numerous clones with hemolytic activity, one clone that produces melanin by a previously unknown mechanism, and one that induces the overproduction of a secondary metabolite native to S. lividans. All bioactive clones were functional in S. lividans but not in E. coli, demonstrating the advantages of screening metagenomic libraries in more than one host.

What is an appropriate reference standard in the quantitation of plaque surface area by intravascular coronary ultrasound?

We reevaluate the predictive accuracy of intravascular ultrasound (IVUS)-derived per cent plaque area stenosis (PAS) in significant coronary lesions (CLs) with or without proximal and distal reference vessel area adjustment. IVUS is valuable in defining moderate CL severity (30 to 70%) in left main (LM) or non-left main (NLM) coronaries using minimum luminal area (MLA) of ≤5.9 and ≤4 mm(2), respectively. Despite a strong correlation with severe CLs, PAS (≥ 70% for NLM and ≥67% for LM) remains underutilized because of confusion about an appropriate reference standard. We studied 120 patients with symptomatic moderate CLs (74 NLM, 46 LM) who underwent IVUS. In-lesion and adjusted PAS were derived by subtracting MLA from in-lesion and proximal or distal reference's external elastic membrane (EEM) area, respectively, divided by corresponding EEM area multiplied by 100. In-lesion PAS was correlated with MLA cutoffs of ≤5.9 and ≤7.5 mm(2) for LM and ≤4 mm(2) for NLM. Adjusted PAS strongly correlated with in-lesion PAS irrespective of reference segment (proximal reference, r = 0.879, p < 0.001; distal reference, r = 0.833, p < 0.001; mean proximal and distal reference, r = 0.896, p < 0.001). Considering MLA of ≤4 mm(2) (for NLM) and ≤5.9 mm(2) (for LM), in-lesion PAS of ≥70 and ≥67%, respectively, explained the majority of severe CLs but the sensitive LM MLA cutoff of ≤7.5 mm(2) showed higher predictive accuracy. Based on results, both in-lesion PAS and adjusted PAS can be used interchangeably and correlate strongly with MLA.

MbtH-like proteins as integral components of bacterial nonribosomal peptide synthetases.

The biosynthesis of many natural products of clinical interest involves large, multidomain enzymes called nonribosomal peptide synthetases (NRPSs). In bacteria, many of the gene clusters coding for NRPSs also code for a member of the MbtH-like protein superfamily, which are small proteins of unknown function. Using MbtH-like proteins from three separate NRPS systems, we show that these proteins copurify with the NRPSs and influence amino acid activation. As a consequence, MbtH-like proteins are integral components of NRPSs.

Theoretical model of errors in micromirror-based three-dimensional particle tracking.

Several recently developed particle-tracking and imaging methods have achieved three-dimensional sensitivity through the introduction of angled micromirrors into the observation volume of an optical microscope. We model the imaging response of such devices and show how the direct and reflected images of a fluorescent particle are affected. In particle-tracking applications, asymmetric image degradation manifests itself as systematic tracking errors. Based on our results, we identify strategies for reducing systematic errors to the 10nm level in practical applications.