GNA13 suppresses proliferation of ER+ breast cancer cells via ERα dependent upregulation of the MYC oncogene.

GNA13 (Gα13) is one of two alpha subunit members of the G12/13 family of heterotrimeric G-proteins which mediate signaling downstream of GPCRs. It is known to be essential for embryonic development and vasculogenesis and has been increasingly shown to be involved in mediating several steps of cancer progression. Recent studies found that Gα13 can function as an oncogene and contributes to progression and metastasis of multiple tumor types, including ovarian, head and neck and prostate cancers. In most cases, Gα12 and Gα13, as closely related α-subunits in the subfamily, have similar cellular roles. However, in recent years their differences in signaling and function have started to emerge. We previously identified that Gα13 drives invasion of Triple Negative Breast Cancer (TNBC) cells in vitro. As a highly heterogenous disease with various well-defined molecular subtypes (ER+ /Her2-, ER+ /Her2+, Her2+, TNBC) and subtype associated outcomes, the function(s) of Gα13 beyond TNBC should be explored. Here, we report the finding that low expression of GNA13 is predictive of poorer survival in breast cancer, which challenges the conventional idea of Gα12/13 being universal oncogenes in solid tumors. Consistently, we found that Gα13 suppresses the proliferation in multiple ER+ breast cancer cell lines (MCF-7, ZR-75-1 and T47D). Loss of GNA13 expression drives cell proliferation, soft-agar colony formation and in vivo tumor formation in an orthotopic xenograft model. To evaluate the mechanism of Gα13 action, we performed RNA-sequencing analysis on these cell lines and found that loss of GNA13 results in the upregulation of MYC signaling pathways in ER+ breast cancer cells. Simultaneous silencing of MYC reversed the proliferative effect from the loss of GNA13, validating the role of MYC in Gα13 regulation of proliferation. Further, we found Gα13 regulates the expression of MYC, at both the transcript and protein level in an ERα dependent manner. Taken together, our study provides the first evidence for a tumor suppressive role for Gα13 in breast cancer cells and demonstrates for the first time the direct involvement of Gα13 in ER-dependent regulation of MYC signaling. With a few exceptions, elevated Gα13 levels are generally considered to be oncogenic, similar to Gα12. This study demonstrates an unexpected tumor suppressive role for Gα13 in ER+ breast cancer via regulation of MYC, suggesting that Gα13 can have subtype-dependent tumor suppressive roles in breast cancer.

A Prospective, Randomized Controlled Multicenter Trial Comparing the Bard Inlay Optima and the Cook Universa Soft Stent.

OBJECTIVE: To determine if the Bard Inlay Optima with its anti-inflammatory pHreecoat stent coating had reduced stent-related symptoms at Week 1 (W1) and Week 3 (W3) post insertion compared to the Cook Universa Soft (CUS) using the validated Ureteral Stent Symptoms Questionnaire (USSQ). METHODS: A prospective, double-blinded, randomized controlled trial was performed on patients receiving unilateral retrograde double-J stents for urolithiasis or pelviureteric junction obstruction at three public Urology services. One hundred forty patients that met inclusion criteria were randomized in a 1:1 ratio to each stent. Primary endpoints were the mean USSQ index scores for the urinary, pain, general, and sexual health domains at W1 and W3. Secondary endpoints were responses to individual USSQ questions, early stent removal, and postoperative opioid use. RESULTS: No significant difference was found between the two stents in terms of index scores for all USSQ domains, early stent removal or postoperative opioid use. The CUS had worse symptom scores at W1 relating to self-reported urinary tract infection symptoms (3.1 ± 1.3 vs 2.6 ± 1.3, P = .05). The CUS was also associated with higher rate of representation to hospital at W1 (n = 10, 16% vs n = 1, 2%, P < .001) and W3 (n = 15, 25% vs n = 3, 5%, P < .001). This did not remain significant when adjusted to site of recruitment (W1 P = .27; W3 P = .22). CONCLUSION: The Bard Inlay Optima's anti-inflammatory pHreecoat stent coating did not translate to any significant difference in overall postoperative symptoms across urinary, pain, general, and sexual health domains.

Narrowing in on urethral strictures.

BACKGROUND: Urethral strictures have affected men for millennia, and they are commonly iatrogenic, idiopathic or traumatic in nature. When left untreated, urethral strictures can devastate the health and function of the urinary tract. For centuries, simple urethral dilation and endoscopic urethrotomy have provided temporary relief. However, in only the past 20 years, urethroplasty has emerged as a curative treatment. OBJECTIVE: The aims of this article are to review the aetiology and clinical manifestations of urethral strictures, evaluate the traditional but temporising treatments for urethral strictures, highlight the relatively recent evolution of urethroplasty and promote awareness of the success of urethroplasty in curing urethral strictures. DISCUSSION: Urethral strictures are an important, but often overlooked, cause of bothersome urinary symptoms in men. In this article, the authors provide a summary of the aetiology, clinical manifestations and recent trends in the management of urethral strictures.

Betacyanin-inhibited biofilm formation of co-culture of Staphylococcus aureus and Pseudomonas aeruginosa on different polymer surfaces.

Staphylococcus aureus and Pseudomonas aeruginosa are bacteria that cause biofilm-associated infections. The aim of this study was to determine the activity of combined betacyanin fractions from Amaranthus dubius (red spinach) and Hylocereus polyrhizus (red pitahaya) against biofilms formed by co-culture of S. aureus and P. aeruginosa on different polymer surfaces. Various formulations containing different concentrations of the betacyanin fractions were investigated for biofilm-inhibiting activity on polystyrene surfaces using crystal violet assay and scanning electron microscopy. A combination of each betacyanin fraction (0.625 mg mL-1) reduced biofilm formation of five S. aureus strains and four P. aeruginosa strains from optical density values of 1.24-3.84 and 1.25-3.52 to 0.81-2.63 and 0.80-1.71, respectively. These combined fractions also significantly inhibited dual-species biofilms by 2.30 and reduced 1.0-1.3 log CFU cm-2 bacterial attachment on polymer surfaces such as polyvinyl chloride, polyethylene, polypropylene and silicone rubber. This study demonstrated an increase in biofilm-inhibiting activity against biofilms formed by two species using combined fractions than that by using single fractions. Betacyanins found in different plants could collectively be used to potentially decrease the risk of biofilm-associated infections caused by these bacteria on hydrophobic polymers.

Outcome of angioembolization for blunt renal trauma in haemodynamically unstable patients: 10-year analysis of Queensland public hospitals.

BACKGROUND: The aim of the study was to evaluate whether angioembolization is an appropriate alternative method for the management of blunt renal trauma in haemodynamically unstable patients. METHODS: A retrospective analysis was conducted from 2002 to 2012 at three tertiary trauma hospitals in the state of Queensland. Patients who had blunt renal trauma and underwent renal angioembolization or had a trauma nephrectomy were identified using patient records and operating theatre and interventional radiology databases. The inclusion criteria were - haemodynamically unstable patients with blunt renal trauma treated with angioembolization, above the age of 16 years. Patients who underwent angioembolization for other causes such as: penetrating renal trauma, post-procedure, renal tumours, renal angiomyolipomas or arteriovenous malformations were excluded. Patients below the age of 16 were also excluded. Post-embolization renal function, blood pressure, morbidity and mortality were analysed using the paired t2 test. RESULTS: A total of 668 renal trauma patients were identified during this period. Sixteen patients underwent angioembolization for blunt renal trauma. Post-procedure renal function normalized without any hypertension with the median follow up being 4 months. Four patients had post-embolization complications including a urinoma, two devascularized kidneys and one ureteric stricture requiring nephrectomy. There was no mortality. CONCLUSION: Selective angioembolization, where feasible, is an alternative method in the management of haemodynamically stable patients with blunt renal trauma maximizing nephron sparing and producing acceptable long-term outcomes with avoidance of the morbidity of trauma nephrectomy. This is the first study that we know of in Australia analysing the outcome of angioembolization for blunt renal trauma.

Alternative splicing and allosteric regulation modulate the chromatin binding of UHRF1.

UHRF1 is an important epigenetic regulator associated with apoptosis and tumour development. It is a multidomain protein that integrates readout of different histone modification states and DNA methylation with enzymatic histone ubiquitylation activity. Emerging evidence indicates that the chromatin-binding and enzymatic modules of UHRF1 do not act in isolation but interplay in a coordinated and regulated manner. Here, we compared two splicing variants (V1, V2) of murine UHRF1 (mUHRF1) with human UHRF1 (hUHRF1). We show that insertion of nine amino acids in a linker region connecting the different TTD and PHD histone modification-binding domains causes distinct H3K9me3-binding behaviour of mUHRF1 V1. Structural analysis suggests that in mUHRF1 V1, in contrast to V2 and hUHRF1, the linker is anchored in a surface groove of the TTD domain, resulting in creation of a coupled TTD-PHD module. This establishes multivalent, synergistic H3-tail binding causing distinct cellular localization and enhanced H3K9me3-nucleosome ubiquitylation activity. In contrast to hUHRF1, H3K9me3-binding of the murine proteins is not allosterically regulated by phosphatidylinositol 5-phosphate that interacts with a separate less-conserved polybasic linker region of the protein. Our results highlight the importance of flexible linkers in regulating multidomain chromatin binding proteins and point to divergent evolution of their regulation.

Exploring the Specificity, Synergy, and Durability of Auditory and Visual Computer Gameplay Transfer Effects in Healthy Older Adults.

OBJECTIVES: To determine whether auditory and visual computer games yield transfer effects that (a) are modality-specific to verbal memory (auditory stimulus presentation) and visual-processing tests, (b) affect working memory and processing speed, (c) are synergistic for combined game-type play, and (d) are durable. METHOD: A Pilot Study (N = 44) assessed visual transfer effects in a two-group pre-post design. The Main Study (N = 151) employed a 2 (visual games: yes, no) × 2 (auditory games: yes, no) × 3 (test session: pretest, post-test, follow-up) design, allowing different training groups to act as active controls for each other. Neuropsychological test scores were aggregated into verbal-memory (auditory presentation), visual-processing, working-memory, and processing-speed indexes. RESULTS: Visual-processing and working-memory pre-post-training change scores were differentially modulated across the four gameplay groups in the main sample, demonstrating transfer effects differing across both active- and passive-control groups. Visual training yielded modality-specific transfer effects in both samples, transfer to working memory in the main sample, and transfer to processing speed in the pilot sample. There were no comparable transfer effects for auditory training. Combined-visual-and-auditory training failed to yield synergistic effects or any significant transfer effects. Visual-processing transfer effects remained significant at follow-up. DISCUSSION: Visual and auditory games differentially modulated transfer effects. Domain-specific visual transfer effects were found at post-test and were durable at follow-up. Visual gameplay holds potential to ameliorate age-related cognitive decline in visual cognition.

When alpha meets beta, mast cells get hyper.

The evolutionary conservation of the catalytically inactive α-tryptase gene has remained a mystery. In this issue of JEM, Le et al. (2019. J. Exp. Med. https://doi.org/10.1084/jem.20190701) unveil the existence of a novel but natural tryptase, heteromeric α/ß-tryptase, a critical mediator of α-tryptase-associated diseases.

Divergent synthesis of 5',7'-difluorinated dihydroxanthene-hemicyanine fused near-infrared fluorophores.

We describe an expedient access to a 5',6',7'-trifluoro dihydroxanthene-hemicyanine fused scaffold in 2 steps and 54% overall yield from the corresponding salicylic aldehyde. A 6'-regioselective nucleophilic aromatic substitution (SNAr) reaction with a wide range of nitrogen, sulfur or selenium nucleophiles then gives access to 16 near-infrared (NIR) fluorophores emitting in the 710-750 nm range. We also report the experimental and theoretical photophysical investigations of these unique optical agents that include the first series of 6'-heavy atom substituted dihydroxanthenes, extending the pool of polyfluorinated markers for biomedical and material applications.

Necrotizing Fasciitis of the Breast: A Case Report and Literature Review.

A 51-year-old woman presented to our facility with an open wound in the left breast. This was associated with a hard, non-mobile, tender lesion palpable underneath. The wound contained central necrosis with surrounding purulent discharge and accompanying erythema. Following radical debridement of the breast down to the pectoralis fascia the patient had a vacuum-assisted closure (VAC) device dressing applied. Histological examination was consistent with necrotizing fasciitis of the breast.

Identification and characterization of antioxidant peptides from hydrolysate of blue-spotted stingray and their stability against thermal, pH and simulated gastrointestinal digestion treatments.

This study was conducted to identify and characterize antioxidant peptides from the alcalase hydrolysate of the blue-spotted stingray. Purification steps guided by ABTS cation radical (ABTS+) scavenging assay and de novo peptide sequencing produced two peptides, WAFAPA (661.3224 Da) and MYPGLA (650.3098 Da). WAFAPA (EC50 = 12.6 µM) had stronger antioxidant activity than glutathione (EC50 = 13.7 µM) and MYPGLA (EC50 = 19.8 µM). Synergism between WAFAPA and MYPGLA was detected. WAFAPA and MYPGLA surpassed carnosine in their ability to suppress H2O2-induced lipid oxidation. The peptides protected plasmid DNA and proteins from Fenton's reagent-induced oxidative damage. Thermal (25-100 °C) and pH 3-11 treatments did not alter antioxidant activity of the peptides. MYPGLA maintained its antioxidant activity after simulated gastrointestinal digestion, whereas WAFAPA showed a partial loss. The two peptides may have potential applications as functional food ingredients or nutraceuticals, whether used singly or in combination.

To stent or not to stent: the use of transanastomotic biliary stents in liver transplantation and patient outcomes.

BACKGROUND: There is no substantial evidence for the use of biliary stents in bile duct reconstruction during liver transplantation. METHOD: A longitudinal, retrospective cohort study was performed to compare biliary complications between stented and non-stented patients between 2011 and 2015 at the Princess Alexandra Hospital, Brisbane, Australia. RESULTS: We found no significant difference in biliary complications between stented and non-stented groups. Stented patients were 3.31 times as likely to require subsequent intervention, mainly in the form of stent removal. CONCLUSION: These results suggest that there is limited benefit in the placement of endobiliary stents in liver transplantation. Given that this was purely an observational study, causality cannot be proven and a prospective cohort trial would be beneficial in further defining these relationships.

Conformational dynamics of the TTD-PHD histone reader module of the UHRF1 epigenetic regulator reveals multiple histone-binding states, allosteric regulation, and druggability.

UHRF1 is a key mediator of inheritance of epigenetic DNA methylation patterns during cell division and is a putative target for cancer therapy. Recent studies indicate that interdomain interactions critically influence UHRF1's chromatin-binding properties, including allosteric regulation of its histone binding. Here, using an integrative approach that combines small angle X-ray scattering, NMR spectroscopy, and molecular dynamics simulations, we characterized the dynamics of the tandem tudor domain-plant homeodomain (TTD-PHD) histone reader module, including its 20-residue interdomain linker. We found that the apo TTD-PHD module in solution comprises a dynamic ensemble of conformers, approximately half of which are compact conformations, with the linker lying in the TTD peptide-binding groove. These compact conformations are amenable to cooperative, high-affinity histone binding. In the remaining conformations, the linker position was in flux, and the reader adopted both extended and compact states. Using a small-molecule fragment screening approach, we identified a compound, 4-benzylpiperidine-1-carboximidamide, that binds to the TTD groove, competes with linker binding, and promotes open TTD-PHD conformations that are less efficient at H3K9me3 binding. Our work reveals a mechanism by which the dynamic TTD-PHD module can be allosterically targeted with small molecules to modulate its histone reader function for therapeutic or experimental purposes.

Case of a strangulated right paraduodenal fossa hernia in a malrotated gut.

We report an unusual case of a strangulated internal hernia resulting from a right paraduodenal fossa hernia (PDH) in the context of bowel malrotation. There are few documented cases of PDHs associated with a concomitant gut malrotation. Emergency laparotomy was performed based on clinical and radiological. Intraoperatively, the proximal jejunum was seen to enter a hernia sac formed by an aberrant duodenojejunal flexure located to the right of the aorta. This was presumed to be a strangulated internal hernia of the paraduodenal recess in a malrotated gut. The hernia neck was widened and the sac obliterated to allow reduction of the contents. On reduction and warming, the insulted small bowel appeared viable and returned to the abdominal cavity without resection.

Synthesis of N,N-Dialkylamino-nor-Dihydroxanthene-Hemicyanine Fused Near-Infrared Fluorophores and Their First Water-Soluble and/or Bioconjugatable Analogues.

The effective synthesis of extended conjugated N,N-dialkylamino-nor-dihydroxanthene-based fluorophores is described from diversely functionalized salicylic aldehydes. The access to these original fluorescent derivatives proceeded in two steps through a one-pot construction of the unusual nor-dihydroxanthene (nor-DHX) scaffold followed by a diversification step providing a wide variety of nor-DHX-hemicyanine fused dyes emitting in the range of 730-790 nm. The versatility of our approach has enabled a further extension to the late-stage introduction of negatively/positively charged polar groups onto their terminal nitrogen heterocyclic subunit, thereby giving access to the first water-soluble and/or bioconjugatable members of this emerging class of NIR fluorophores. Our water-solubilizing method is easily implementable, and the nor-DHX-hemicyanine skeleton maintains satisfying fluorescence quantum yields (5-20 %) under physiological conditions. Finally, the bioconjugation ability of fluorescent derivatives bearing a free carboxylic acid was demonstrated through the covalent labeling of a model protein, namely, bovine serum albumin.

Divergent Synthesis of Dihydroxanthene-Hemicyanine Fused Near-Infrared Fluorophores through the Late-Stage Amination of a Bifunctional Precursor.

A late-stage amination of a bifunctional dihydroxanthene (DHX) scaffold is reported to access a wide variety of new near-infrared (NIR) chromophores/fluorophores. The divergent approach allows the coupling of aliphatic and aromatic amines and readily provides molecular diversity shedding light on the structure-fluorescence relationship of this emerging class of NIR fluorophores.

Accessibility of different histone H3-binding domains of UHRF1 is allosterically regulated by phosphatidylinositol 5-phosphate.

UHRF1 is a multidomain protein crucially linking histone H3 modification states and DNA methylation. While the interaction properties of its specific domains are well characterized, little is known about the regulation of these functionalities. We show that UHRF1 exists in distinct active states, binding either unmodified H3 or the H3 lysine 9 trimethylation (H3K9me3) modification. A polybasic region (PBR) in the C terminus blocks interaction of a tandem tudor domain (TTD) with H3K9me3 by occupying an essential peptide-binding groove. In this state the plant homeodomain (PHD) mediates interaction with the extreme N terminus of the unmodified H3 tail. Binding of the phosphatidylinositol phosphate PI5P to the PBR of UHRF1 results in a conformational rearrangement of the domains, allowing the TTD to bind H3K9me3. Our results define an allosteric mechanism controlling heterochromatin association of an essential regulatory protein of epigenetic states and identify a functional role for enigmatic nuclear phosphatidylinositol phosphates.

Stabilized Wittig olefination for bioconjugation.

Stabilized Wittig olefination holds great potential as a bioconjugation reaction. We demonstrate that the reaction of stabilized phosphorus ylides (or phosphonium salts) with aryl aldehydes is sufficiently robust to be used for live cell affinity isolation and fluorescence tagging of a protein, FKBP12.

Multivalent histone engagement by the linked tandem Tudor and PHD domains of UHRF1 is required for the epigenetic inheritance of DNA methylation.

Histone post-translational modifications regulate chromatin structure and function largely through interactions with effector proteins that often contain multiple histone-binding domains. While significant progress has been made characterizing individual effector domains, the role of paired domains and how they function in a combinatorial fashion within chromatin are poorly defined. Here we show that the linked tandem Tudor and plant homeodomain (PHD) of UHRF1 (ubiquitin-like PHD and RING finger domain-containing protein 1) operates as a functional unit in cells, providing a defined combinatorial readout of a heterochromatin signature within a single histone H3 tail that is essential for UHRF1-directed epigenetic inheritance of DNA methylation. These findings provide critical support for the "histone code" hypothesis, demonstrating that multivalent histone engagement plays a key role in driving a fundamental downstream biological event in chromatin.

The psychosocial impact of child domestic work: a study from India and the Philippines.

The aim of the study was to explore the effects of domestic employment on the well-being of child domestic workers (CDWs) in India and the Philippines. A questionnaire was administered to 700 CDWs and 700 school-attending controls in the two countries. In India, 36% of CDWs started work before age 12, 48% worked because of poverty or to repay loans, 46% worked >10 h per day, and 31% were physically punished by employers. Filipino CDWs were mainly migrants from rural areas, 47% were working to continue their studies and 87% were attending school, compared with 35% of Indians. In India, 67% of CDWs and 25% of controls scored in the lowest tertile (p<0.001) compared with 36% and 30%, respectively, in the Philippines (p=02). Key significant correlates of low psychosocial scores were non-attendance at school, long working hours, physical punishment, limited support networks and poor health. This study shows that it is not domestic work that is intrinsically harmful, but rather the circumstances and conditions of work, which could be improved through pragmatic regulatory measures.