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The article discusses studies concerning the automated design for RNA devices. It says that researchers V. K. Mutalik and colleagues have created a computer-aided design (CAD) tool which can design a large regulatory RNA molecule library using an IS10 insertion sequence system. Furthermore, researchers J. M. Carothers and colleagues have combined biophysical kinetic RNA folding and biochemical model simulations to design RNA devices which can control Escherichia (E.) Coli gene expression.

The injuries sustained during one season by players at an Ulster Senior Rugby Club were documented with reference to time of injury, phase of play, team position, and nature of injury. The overall injury pattern in Irish rugby as found in this study is broadly comparable with that in similar studies in England and Scotland. Eighty-four players were injured in total. Three of these had fractures of which one required hospital admission for open reduction of a fractured wrist. The remaining eighty-one players had various types and grades of soft-tissue damage. The tackle accounted for approximately one-third of all injuries. There were few serious injuries and none involving the spine. Injuries to forwards made up almost sixty per cent of the total. [ABSTRACT FROM AUTHOR]

OC19.05: Evaluating cut-off values for progesterone, single hCG and hCG ratio to define pregnancy viability and location in women with a pregnancy of unknown location (PUL) We aimed to define the performance of single measurements of serum hCG, progesterone and the hCG ratio (hCG 48hrs/hCG 0hrs) for diagnosing pregnancy viability and location. We found 19.6% VIUP when single hCG >1000 IU/L, 14.4% when single hCG >2000 IU/L and 11.4% when single hCG >3000 IU/L. No single hCG cut-off excludes an EP. [Extracted from the article]

OC01.04: A multicentre trial on the performance of a two-step triage protocol based on initial serum progesterone and serial hCG used to manage pregnancies of unknown location (PUL) We previously published a two-step triage protocol based on the presenting serum progesterone (step 1) and M6 model, a logistic regression model based on the initial hCG and hCG ratio two days later (step 2) to select PUL at high-risk of ectopic pregnancy (EP). In seven centres, if the progesterone was <=2nmol/l, patients were discharged with a follow-up urine pregnancy test in two weeks to confirm a negative result. [Extracted from the article]

Objective: The M6 risk-prediction model was published as part of a two-step protocol using an initial progesterone level of ≤ 2 nmol/L to identify probable failing pregnancies (Step 1) followed by the M6 model (Step 2). The M6 model has been shown to have good triage performance for stratifying women with a pregnancy of unknown location (PUL) as being at low or high risk of harboring an ectopic pregnancy (EP). This study validated the triage performance of the two-step protocol in clinical practice by evaluating the number of protocol-related adverse events and how effectively patients were triaged.Methods: This was a prospective multicenter interventional study of 3272 women with a PUL, carried out between January 2015 and January 2017 in four district general hospitals and four university teaching hospitals in the UK. The final pregnancy outcome was defined as: a failed PUL (FPUL), an intrauterine pregnancy (IUP) or an EP (including persistent PUL (PPUL)). FPUL and IUP were grouped as low-risk and EP/PPUL as high-risk PUL. Serum progesterone and human chorionic gonadotropin (hCG) levels were measured at presentation in all patients. If the initial progesterone level was ≤ 2 nmol/L, patients were discharged and were asked to have a follow-up urine pregnancy test in 2 weeks to confirm a negative result. If the progesterone level was > 2 nmol/L or a measurement had not been taken, hCG level was measured again at 48 h and results were entered into the M6 model. Patients were managed according to the outcome predicted by the protocol. Those classified as 'low risk, probable FPUL' were advised to perform a urine pregnancy test in 2 weeks and those classified as 'low risk, probable IUP' were invited for a scan a week later. When a woman with a PUL was classified as high risk (i.e. risk of EP ≥ 5%) she was reviewed clinically within 48 h. One center used a progesterone cut-off of ≤ 10 nmol/L and its data were analyzed separately. If the recommended management protocol was not adhered to, this was recorded as a protocol deviation and classified as: unscheduled visit for clinician reason, unscheduled visit for patient reason or incorrect timing of blood test or ultrasound scan. The classifications outlined in the UK Good Clinical Practice (GCP) guidelines were used to evaluate the incidence of adverse events. Data were analyzed using descriptive statistics.Results: Of the 3272 women with a PUL, 2625 were included in the final analysis (317 met the exclusion criteria or were lost to follow-up, while 330 were evaluated using a progesterone cut-off of ≤ 10 nmol/L). Initial progesterone results were available for 2392 (91.1%) patients. In Step 1, 407 (15.5%) patients were classified as low risk (progesterone ≤ 2 nmol/L), of whom seven (1.7%) were ultimately diagnosed with an EP. In 279 of the remaining 2218 women with a PUL, the M6 model was not applied owing to protocol deviation or because the outcome was already known (usually on the basis of an ultrasound scan) before a second hCG reading was taken; of these patients, 30 were diagnosed with an EP. In Step 2, 1038 women with a PUL were classified as low risk, of whom eight (0.8%) had a final outcome of EP. Of 901 women classified as high risk at Step 2, 275 (30.5%) had an EP. Therefore, 275/320 (85.9%) EPs were correctly classified as high risk. Overall, 1445/2625 PUL (55.0%) were classified as low risk, of which 15 (1.0%) were EP. None of these cases resulted in a ruptured EP or significant clinical harm. Sixty-two women participating in the study had an adverse event, but no woman had a serious adverse event as defined in the UK GCP guidelines.Conclusions: This study has shown that the two-step protocol incorporating the M6 model effectively triaged the majority of women with a PUL as being at low risk of an EP, minimizing the follow-up required for these patients after just two visits. There were few misclassified EPs and none of these women came to significant clinical harm or suffered a serious adverse clinical event. The two-step protocol incorporating the M6 model is an effective and clinically safe way of rationalizing the management of women with a PUL. Copyright © 2019 ISUOG. Published by John Wiley & Sons Ltd. [ABSTRACT FROM AUTHOR]

An abstract of the article "A multicentre trial on the performance and complications of a two-step triage protocol based on initial serum progesterone and serial serum hCG to manage pregnancy of unknown location (PUL)," by S. Bobdiwala and colleagues is presented.

A correction is presented to the article "Recoded organisms engineered to depend on synthetic amino acids" by Alexis J. Rovner, Adrian D. Haimovich, Spencer R. Katz, and others in volume 518 of the journal, noting financial interests related to a patent application.

Engineering of the translation apparatus has permitted the site-specific incorporation of nonstandard amino acids (nsAAs) into proteins, thereby expanding the genetic code of organisms. Conventional approaches have focused on porting tRNAs and aminoacyl-tRNA synthetases (aaRS) from archaea into bacterial and eukaryotic systems where they have been engineered to site-specifically encode nsAAs. More recent work in genome engineering has opened up the possibilities of whole genome recoding, in which organisms with alternative genetic codes have been constructed whereby codons removed from the genetic code can be repurposed as new sense codons dedicated for incorporation of nsAAs. These advances, together with the advent of engineered ribosomes and new molecular evolution methods, enable multisite incorporation of nsAAs and nonstandard monomers (nsM) paving the way for the template-directed production of functionalized proteins, new classes of polymers, and genetically encoded materials. [ABSTRACT FROM AUTHOR]

Background: Psychological illness occurring in association with hyperemesis gravidarum (HG) has been widely reported.Objective: To determine if there is a higher incidence of psychological morbidity in women with HG compared with women without significant nausea and vomiting in pregnancy.Search Strategy: PubMed, MEDLINE, Embase and PsychINFO were searched up to September 2015.Selection Criteria: Articles referring to psychological morbidity in relation to HG. For meta-analysis case-control studies using numerical scales to compare psychological symptoms.Data Collection and Analysis: Articles were independently assessed for inclusion by two reviewers and methodology was appraised using the Newcastle Ottawa Scale. Comparison was made using the standard mean difference (SMD) in symptom scale scores.Main Results: In all, 59 articles were included in the systematic review, 12 of these were used in the meta-analysis. Meta-analysis of depression scale scores demonstrated a very large effect with statistically significantly higher depression scale scores in women with HG (SMD 1.22; 95% CI 0.80-1.64; P ≤ 0.01) compared with controls. Meta-analysis of anxiety scores demonstrated a large effect with statistically significantly higher anxiety disorder scale scores in women with HG (SMD 0.86; 95% CI 0.53-1.19; P ≤ 0.01). In both analyses significant heterogeneity was identified (depression and HG I2  = 94%, P ≤ 0.01; anxiety and HG I2  = 84%, P = 0.02).Conclusions: Our systematic review and meta-analysis have shown a significantly increased frequency of depression and anxiety in women with HG. The findings should prompt service development for women with HG that includes provision of psychological care and support.Tweetable Abstract: Meta-analysis demonstrates an increase in #PsychologicalMorbidity in women with #HyperemesisGravidarum. [ABSTRACT FROM AUTHOR]

The development of biomolecular devices that interface with biological systems to reveal new insights and produce novel functions is one of the defining goals of synthetic biology. Our lab previously described a synthetic, riboregulator system that affords for modular, tunable, and tight control of gene expression in vivo. Here we highlight several experimental advantages unique to this RNA-based system, including physiologically relevant protein production, component modularity, leakage minimization, rapid response time, tunable gene expression, and independent regulation of multiple genes. We demonstrate this utility in four sets of in vivo experiments with various microbial systems. Specifically, we show that the synthetic riboregulator is well suited for GFP fusion protein tracking in wild- type cells, tight regulation of toxic protein expression, and sensitive perturbation of stress response networks. We also show that the system can be used for logic-based computing of multiple, orthogonal inputs, resulting in the development of a programmable kill switch for bacteria. This work establishes a broad, easy-to-use synthetic biology platform for microbiology experiments and biotechnology applications. [ABSTRACT FROM AUTHOR]

The use of biologics in the treatment of numerous diseases has increased steadily over the past decade due to their high specificities, low toxicity, and limited side effects. Despite this success, peptide- and protein-based drugs are limited by short half-lives and immunogenicity. To address these challenges, we use a genomically recoded organism to produce genetically encoded elastin-like polypeptide-protein fusions containing multiple instances of para-azidophenylalanine (pAzF). Precise lipidation of these pAzF residues generated a set of sequence-defined synthetic biopolymers with programmable binding affinity to albumin without ablating the activity of model fusion proteins, and with tunable blood serum half-lives spanning 5 to 94% of albumin's half-life in a mouse model. Our findings present a proof of concept for the use of genetically encoded bioorthogonal conjugation sites for multisite lipidation to tune protein stability in mouse serum. This work establishes a programmable approach to extend and tune the half-life of protein or peptide therapeutics and a technical foundation to produce functionalized biopolymers endowed with programmable chemical and biophysical properties with broad applications in medicine, materials science, and biotechnology. [ABSTRACT FROM AUTHOR]

The Emirates Exploration Imager (EXI) on-board the Emirates Mars Mission (EMM) offers both regional and global imaging capabilities for studies of the Martian atmosphere. EXI is a framing camera with a field-of-view (FOV) that will easily capture the martian disk at the EMM science orbit periapsis. EXI provides 6 bandpasses nominally centered on 220, 260, 320, 437, 546, 635 nm using two telescopes (ultraviolet (UV) and visible(VIS)) with separate optics and detectors. Images of the full-disk are acquired with a resolution of 2–4 km per pixel, where the variation is driven by periapsis and apoapsis points of the orbit, respectively. By combining multiple observations within an orbit with planetary rotation, EXI is able to provide diurnal sampling over most of the planet on the scale of 10 days. As a result, the EXI dataset allows for the delineation of diurnal and seasonal timescales in the behavior of atmospheric constituents such as water ice clouds and ozone. This combination of temporal and spatial distinguishes EXI from somewhat similar imaging systems, including the Mars Color Imager (MARCI) onboard the Mars Reconnaissance Orbiter (MRO) (Malin et al. in Icarus 194(2):501–512, 2008) and the various cameras on-board the Hubble Space Telescope (HST; e.g., James et al. in J. Geophys. Res. 101(E8):18,883–18,890, 1996; Wolff et al. in J. Geophys. Res. 104(E4):9027–9042, 1999). The former, which has comparable spatial and spectral coverage, possesses a limited local time view (e.g., mid-afternoon). The latter, which provides full-disk imaging, has limited spatial resolution through most of the Martian year and is only able to provide (at most) a few observations per year given its role as a dedicated, queue-based astrophysical observatory. In addition to these unique attributes of the EXI observations, the similarities with other missions allows for the leveraging of both past and concurrent observations. For example, with MARCI, one can build on the ∼6 Mars years of daily global UV images as well as those taken concurrently with EXI. [ABSTRACT FROM AUTHOR]

Objective: To validate externally five approaches to predict ectopic pregnancy (EP) in pregnancies of unknown location (PUL): the M6P and M6NP risk models, the two‐step triage strategy (2ST, which incorporates M6P), the M4 risk model, and beta human chorionic gonadotropin ratio cut‐offs (BhCG‐RC). Design: Secondary analysis of a prospective cohort study. Setting: Eight UK early pregnancy assessment units. Population: Women presenting with a PUL and BhCG >25 IU/l. Methods: Women were managed using the 2ST protocol: PUL were classified as low risk of EP if presenting progesterone ≤2 nmol/l; the remaining cases returned 2 days later for triage based on M6P. EP risk ≥5% was used to classify PUL as high risk. Missing values were imputed, and predictions for the five approaches were calculated post hoc. We meta‐analysed centre‐specific results. Main outcome measures: Discrimination, calibration and clinical utility (decision curve analysis) for predicting EP. Results: Of 2899 eligible women, the primary analysis excluded 297 (10%) women who were lost to follow up. The area under the ROC curve for EP was 0.89 (95% CI 0.86–0.91) for M6P, 0.88 (0.86–0.90) for 2ST, 0.86 (0.83–0.88) for M6NP and 0.82 (0.78–0.85) for M4. Sensitivities for EP were 96% (M6P), 94% (2ST), 92% (N6NP), 80% (M4) and 58% (BhCG‐RC); false‐positive rates were 35%, 33%, 39%, 24% and 13%. M6P and 2ST had the best clinical utility and good overall calibration, with modest variability between centres. Conclusions: 2ST and M6P performed best for prediction and triage in PUL. The M6 model, as part of a two‐step triage strategy, is the best approach to characterise and triage PULs. The M6 model, as part of a two‐step triage strategy, is the best approach to characterise and triage PULs. [ABSTRACT FROM AUTHOR]

Objective: To assess whether sonographic diagnosis of intrauterine hematoma (IUH) in the first trimester of pregnancy is associated with first-trimester miscarriage and antenatal, delivery and neonatal complications.Methods: This was a prospective observational cohort study of women with an intrauterine singleton pregnancy between 5 and 14 weeks' gestation recruited at Queen Charlotte's and Chelsea Hospital, London, UK, between March 2014 and March 2016. Participants underwent serial ultrasound examinations in the first trimester, and the presence, location, size and persistence of any IUH was evaluated. First-trimester miscarriage was defined as pregnancy loss before 14 weeks' gestation. Clinical symptoms, including pelvic pain and vaginal bleeding, were recorded at each visit using validated symptom scores. Antenatal, delivery and neonatal outcomes were obtained from hospital records. Logistic regression analysis and the chi-square test were used to assess the association between the presence and features of IUH and the incidence of adverse pregnancy outcome. Odds ratios (OR) were first adjusted for maternal age (aOR) and then further adjusted for the presence of vaginal bleeding or pelvic pain in the first trimester.Results: Of 1003 women recruited to the study, 946 were included in the final analysis and of these, 268 (28.3%) were diagnosed with an IUH in the first trimester. The presence of IUH was associated with the incidence of preterm birth (aOR, 1.94 (95% CI, 1.07-3.52)), but no other individual or overall antenatal, delivery or neonatal complications. No association was found between the presence of IUH in the first trimester and first-trimester miscarriage (aOR, 0.81 (95% CI, 0.44-1.50)). These findings were independent of the absolute size of the hematoma and the presence of vaginal bleeding or pelvic pain in the first trimester. When IUH was present in the first trimester, there was no association between its size, content or position in relation to the gestational sac and overall antenatal, delivery and neonatal complications. Diagnosis of a retroplacental IUH was associated with an increased risk of overall antenatal complications (P = 0.04).Conclusions: Our findings demonstrate that there is no association between the presence of IUH in the first trimester and first-trimester miscarriage. However, an association with preterm birth, independently of the presence of symptoms of pelvic pain and/or vaginal bleeding, is evident. Women diagnosed with IUH in the first trimester should be counseled about their increased risk of preterm birth and possibly be offered increased surveillance during the course of their pregnancy. Copyright © 2019 ISUOG. Published by John Wiley & Sons Ltd. [ABSTRACT FROM AUTHOR]

The genetic code defines how information in the genome is translated into protein. Aside from a handful of isolated exceptions, this code is universal. Researchers have developed techniques to artificially expand the genetic code, repurposing codons and translational machinery to incorporate nonstandard amino acids (nsAAs) into proteins. A key challenge for robust genetic code expansion is orthogonality; the engineered machinery used to introduce nsAAs into proteins must co-exist with native translation and gene expression without cross-reactivity or pleiotropy. The issue of orthogonality manifests at several levels, including those of codons, ribosomes, aminoacyl-tRNA synthetases, tRNAs, and elongation factors. In this concept paper, we describe advances in genome recoding, translational engineering and associated challenges rooted in establishing orthogonality needed to expand the genetic code. [ABSTRACT FROM AUTHOR]

Objective: To assess prospectively the association between pelvic pain, vaginal bleeding, and nausea and vomiting occurring in the first trimester of pregnancy and the incidence of later adverse pregnancy outcomes.Methods: This was a prospective observational cohort study of consecutive women with confirmed intrauterine singleton pregnancy between 5 and 14 weeks' gestation recruited at Queen Charlotte's & Chelsea Hospital, London, UK, from March 2014 to March 2016. Serial ultrasound scans were performed in the first trimester. Participants completed validated symptom scores for vaginal bleeding, pelvic pain, and nausea and vomiting. The key symptom of interest was any pelvic pain and/or vaginal bleeding during the first trimester. Pregnancies were followed up until the final outcome was known. Antenatal, delivery and neonatal outcomes were obtained from hospital records. Logistic regression analysis was used to assess the association between first-trimester symptoms and pregnancy complications by calculating adjusted odds ratios (aOR) with correction for maternal age.Results: Of 1003 women recruited, 847 pregnancies were included in the final analysis following exclusion of cases due to first-trimester miscarriage (n = 99), termination of pregnancy (n = 20), loss to follow-up (n = 32) or withdrawal from the study (n = 5). Adverse antenatal complications were observed in 166/645 (26%) women with pelvic pain and/or vaginal bleeding in the first trimester (aOR = 1.79; 95% CI, 1.17-2.76) and in 30/181 (17%) women with no symptoms. Neonatal complications were observed in 66/634 (10%) women with and 11/176 (6%) without pelvic pain and/or vaginal bleeding (aOR = 1.73; 95% CI, 0.89-3.36). Delivery complications were observed in 402/615 (65%) women with and 110/174 (63%) without pelvic pain and/or vaginal bleeding during the first trimester (aOR = 1.16; 95% CI, 0.81-1.65). For 18 of 20 individual antenatal complications evaluated, incidence was higher among women with pelvic pain and/or vaginal bleeding, despite the overall incidences being low. Nausea and vomiting in pregnancy showed little association with adverse pregnancy outcomes.Conclusions: Our study suggests that there is an increased incidence of antenatal complications in women experiencing pelvic pain and/or vaginal bleeding in the first trimester. This should be considered when advising women attending early-pregnancy units. Copyright © 2019 ISUOG. Published by John Wiley & Sons Ltd. [ABSTRACT FROM AUTHOR]

Post-translational phosphorylation is essential to human cellular processes, but the transient, heterogeneous nature of this modification complicates its study in native systems. We developed an approach to interrogate phosphorylation and its role in protein-protein interactions on a proteome-wide scale. We genetically encoded phosphoserine in recoded E. coli and generated a peptide-based heterologous representation of the human serine phosphoproteome. We designed a single-plasmid library encoding >100,000 human phosphopeptides and confirmed the site-specific incorporation of phosphoserine in >36,000 of these peptides. We then integrated our phosphopeptide library into an approach known as Hi-P to enable proteome-level screens for serine-phosphorylation-dependent human protein interactions. Using Hi-P, we found hundreds of known and potentially new phosphoserine-dependent interactors with 14-3-3 proteins and WW domains. These phosphosites retained important binding characteristics of the native human phosphoproteome, as determined by motif analysis and pull-downs using full-length phosphoproteins. This technology can be used to interrogate user-defined phosphoproteomes in any organism, tissue, or disease of interest. [ABSTRACT FROM AUTHOR]

The degeneracy of the genetic code allows nucleic acids to encode amino acid identity as well as noncoding information for gene regulation and genome maintenance. The rare arginine codons AGA and AGG (AGR) present a case study in codon choice, with AGRs encoding important transcriptional and translational properties distinct from the other synonymous alternatives (CGN). We created a strain of Escherichia coli with all 123 instances of AGR codons removed from all essential genes. We readily replaced 110 AGR codons with the synonymous CGU codons, but the remaining 13 "recalcitrant" AGRs required diversification to identify viable alternatives. Successful replacement codons tended to conserve local ribosomal binding site-like motifs and local mRNA secondary structure, sometimes at the expense of amino acid identity. Based on these observations, we empirically defined metrics for a multidimensional "safe replacement zone" (SRZ) within which alternative codons are more likely to be viable. To evaluate synonymous and nonsynonymous alternatives to essential AGRs further, we implemented a CRISPR/Cas9-based method to deplete a diversified population of a wild-type allele, allowing us to evaluate exhaustively the fitness impact of all 64 codon alternatives. Using this method, we confirmed the relevance of the SRZ by tracking codon fitness over time in 14 different genes, finding that codons that fall outside the SRZ are rapidly depleted from a growing population. Our unbiased and systematic strategy for identifying unpredicted design flaws in synthetic genomes and for elucidating rules governing codon choice will be crucial for designing genomes exhibiting radically altered genetic codes. [ABSTRACT FROM AUTHOR]

Genetically modified organisms (GMOs) are increasingly used in research and industrial systems to produce high-value pharmaceuticals, fuels and chemicals. Genetic isolation and intrinsic biocontainment would provide essential biosafety measures to secure these closed systems and enable safe applications of GMOs in open systems, which include bioremediation and probiotics. Although safeguards have been designed to control cell growth by essential gene regulation, inducible toxin switches and engineered auxotrophies, these approaches are compromised by cross-feeding of essential metabolites, leaked expression of essential genes, or genetic mutations. Here we describe the construction of a series of genomically recoded organisms (GROs) whose growth is restricted by the expression of multiple essential genes that depend on exogenously supplied synthetic amino acids (sAAs). We introduced a Methanocaldococcus jannaschii tRNA:aminoacyl-tRNA synthetase pair into the chromosome of a GRO derived from Escherichia coli that lacks all TAG codons and release factor 1, endowing this organism with the orthogonal translational components to convert TAG into a dedicated sense codon for sAAs. Using multiplex automated genome engineering, we introduced in-frame TAG codons into 22 essential genes, linking their expression to the incorporation of synthetic phenylalanine-derived amino acids. Of the 60 sAA-dependent variants isolated, a notable strain harbouring three TAG codons in conserved functional residues of MurG, DnaA and SerS and containing targeted tRNA deletions maintained robust growth and exhibited undetectable escape frequencies upon culturing ∼1011 cells on solid media for 7 days or in liquid media for 20 days. This is a significant improvement over existing biocontainment approaches. We constructed synthetic auxotrophs dependent on sAAs that were not rescued by cross-feeding in environmental growth assays. These auxotrophic GROs possess alternative genetic codes that impart genetic isolation by impeding horizontal gene transfer and now depend on the use of synthetic biochemical building blocks, advancing orthogonal barriers between engineered organisms and the environment. [ABSTRACT FROM AUTHOR]

The site-specific incorporation of nonstandard amino acids (nsAAs) during translation has expanded the chemistry and function of proteins. The nsAA para -azido-phenylalanine (pAzF) encodes a biorthogonal chemical moiety that facilitates "click" reactions to attach diverse chemical groups for protein functionalization. However, the azide moiety is unstable in physiological conditions and is reduced to para -amino-phenylalanine (pAF). Azide reduction decreases the yield of pAzF residues in proteins to 50%–60% per azide and limits protein functionalization by click reactions. Here, we describe the use of a pH-tunable diazotransfer reaction that converts pAF to pAzF at >95% efficiency in proteins. The method selectively restores pAzF at multiple sites per protein without introducing off-target modifications. This work addresses a key limitation in the production of pAzF-containing proteins by restoring azides for multi-site functionalization with diverse chemical moieties, setting the stage for the production of genetically encoded biomaterials with broad applications in biotherapeutics, materials science, and biotechnology. [Display omitted] • In vivo reduction of para -azido-phenylalanine (pAzF) hinders protein functionalization • Reaction with imidazole-1-sulfonyl azide selectively recovers pAzF • Recovery of pAzF in proteins enables robust multi-site functionalization Protein expression with para -azidophenylalanine (pAzF) enables a site-specific modification with synthetic modalities to endow proteins and biopolymers with new functionalities. The azide can be intracellularly reduced to an amine, preventing homogeneous functionalization of these proteins. Here, Arranz-Gibert et al. develop a strategy to selectively restore pAzF in proteins for robust functionalization. [ABSTRACT FROM AUTHOR]

Small molecules encoded by biosynthetic pathways mediate cross-species interactions and harbor untapped potential, which has provided valuable compounds for medicine and biotechnology. Since studying biosynthetic gene clusters in their native context is often difficult, alternative efforts rely on heterologous expression, which is limited by host-specific metabolic capacity and regulation. Here, we describe a computational-experimental technology to redesign genes and their regulatory regions with hybrid elements for cross-species expression in Gram-negative and -positive bacteria and eukaryotes, decoupling biosynthetic capacity from host-range constraints to activate silenced pathways. These synthetic genetic elements enabled the discovery of a class of microbiome-derived nucleotide metabolites—tyrocitabines—from Lactobacillus iners. Tyrocitabines feature a remarkable orthoester-phosphate, inhibit translational activity, and invoke unexpected biosynthetic machinery, including a class of "Amadori synthases" and "abortive" tRNA synthetases. Our approach establishes a general strategy for the redesign, expression, mobilization, and characterization of genetic elements in diverse organisms and communities. [Display omitted] • Redesign of multigene pathways for expression in prokaryotes and eukaryotes • Development and mobilization of hybrid cross-kingdom expression technology • Discovery of tyrocitabines, a class of nucleotide metabolites from the human microbiome • Tyrocitabine biosynthesis invokes unusual tRNA synthetase-mediated orthoester formation A computational and experimental strategy was developed to redesign biosynthetic gene clusters into synthetic genetic elements that can be expressed across a wide range of prokaryote and eukaryote hosts, expanding our ability to discover and characterize a variety of natural products and biosynthetic pathways. [ABSTRACT FROM AUTHOR]

We describe the construction and characterization of a genomically recoded organism (GRO). We replaced all known UAG stop codons in Escherichia coli MG1655 with synonymous UAA codons, which permitted the deletion of release factor 1 and reassignment of UAG translation function. This GRO exhibited improved properties for incorporation of nonstandard amino acids that expand the chemical diversity of proteins in vivo. The GRO also exhibited increased resistance to T7 bacteriophage, demonstrating that new genetic codes could enable increased viral resistance. [ABSTRACT FROM AUTHOR]

Risk prediction models utilising hCG and progesterone are effective tools to manage PUL pending a final diagnosis by TVUS. The M6 prediction model uses progesterone, hCG and the hCG ratio (hCG 48 hours/hCG 0 hours). We assessed the effectiveness of PUL risk stratification at 96 hours using the M6 model and the time taken to confirm a diagnosis of ectopic pregnancy using TVUS. [Extracted from the article]

Intrauterine hematoma's (IUH) are commonly seen in the first trimester. Pregnancy outcomes were first trimester miscarriage or the development of antenatal, delivery or neonatal complications. There was a relationship between IUH and preterm birth (PTB) (OR 1.94; 95% CI 1.07-3.52) and PTB-Preterm Premature Rupture of Membranes (PPROM) (OR 1.84; 95% CI 1.03-3.28) when these were grouped together, irrespective of IUH size. [Extracted from the article]

To externally validate the performance of the M6 risk model, and the two step triage system (2ST) to identify pregnancies of unknown location (PUL) at high risk of ectopic pregnancy. Step 1 triages patients as low risk if presenting progesterone (if available) is <=2 nmol/L, else a step 2 classification of low or high risk by M6 is calculated. We excluded women with presenting hCG <=25 IU/L. One centre performed step 1 triage using presenting progesterone <=10 nmol/L. PUL outcome was defined as failed (FPUL), intra-uterine (IUP), or ectopic/persisting (EP). [Extracted from the article]

The article proposes the Human Genome Project-Write (HGP-Write) and highlights the need for technology and an ethical framework for genome-scale engineering. Topics discussed include the importance of responsible innovation and some controversial aspects of the genome project. The potential applications of HGP-Write are also cited including transplantable human organs.