APOE3ch alters microglial response and suppresses Aß-induced tau seeding and spread.

A recent case report described an individual who was a homozygous carrier of the APOE3 Christchurch (APOE3ch) mutation and resistant to autosomal dominant Alzheimer's Disease (AD) caused by a PSEN1-E280A mutation. Whether APOE3ch contributed to the protective effect remains unclear. We generated a humanized APOE3ch knock-in mouse and crossed it to an amyloid-ß (Aß) plaque-depositing model. We injected AD-tau brain extract to investigate tau seeding and spreading in the presence or absence of amyloid. Similar to the case report, APOE3ch expression resulted in peripheral dyslipidemia and a marked reduction in plaque-associated tau pathology. Additionally, we observed decreased amyloid response and enhanced microglial response around plaques. We also demonstrate increased myeloid cell phagocytosis and degradation of tau aggregates linked to weaker APOE3ch binding to heparin sulfate proteoglycans. APOE3ch influences the microglial response to Aß plaques, which suppresses Aß-induced tau seeding and spreading. The results reveal new possibilities to target Aß-induced tauopathy.

Lactate Metabolism in Human Lung Tumors.

Cancer cells consume glucose and secrete lactate in culture. It is unknown whether lactate contributes to energy metabolism in living tumors. We previously reported that human non-small-cell lung cancers (NSCLCs) oxidize glucose in the tricarboxylic acid (TCA) cycle. Here, we show that lactate is also a TCA cycle carbon source for NSCLC. In human NSCLC, evidence of lactate utilization was most apparent in tumors with high 18fluorodeoxyglucose uptake and aggressive oncological behavior. Infusing human NSCLC patients with 13C-lactate revealed extensive labeling of TCA cycle metabolites. In mice, deleting monocarboxylate transporter-1 (MCT1) from tumor cells eliminated lactate-dependent metabolite labeling, confirming tumor-cell-autonomous lactate uptake. Strikingly, directly comparing lactate and glucose metabolism in vivo indicated that lactate's contribution to the TCA cycle predominates. The data indicate that tumors, including bona fide human NSCLC, can use lactate as a fuel in vivo.

Tree islands enhance biodiversity and functioning in oil palm landscapes.

In the United Nations Decade on Ecosystem Restoration1, large knowledge gaps persist on how to increase biodiversity and ecosystem functioning in cash crop-dominated tropical landscapes2. Here, we present findings from a large-scale, 5-year ecosystem restoration experiment in an oil palm landscape enriched with 52 tree islands, encompassing assessments of ten indicators of biodiversity and 19 indicators of ecosystem functioning. Overall, indicators of biodiversity and ecosystem functioning, as well as multidiversity and ecosystem multifunctionality, were higher in tree islands compared to conventionally managed oil palm. Larger tree islands led to larger gains in multidiversity through changes in vegetation structure. Furthermore, tree enrichment did not decrease landscape-scale oil palm yield. Our results demonstrate that enriching oil palm-dominated landscapes with tree islands is a promising ecological restoration strategy, yet should not replace the protection of remaining forests.

Loop extrusion as a mechanism for formation of DNA damage repair foci.

The repair of DNA double-strand breaks (DSBs) is essential for safeguarding genome integrity. When a DSB forms, the PI3K-related ATM kinase rapidly triggers the establishment of megabase-sized, chromatin domains decorated with phosphorylated histone H2AX (γH2AX), which act as seeds for the formation of DNA-damage response foci1. It is unclear how these foci are rapidly assembled to establish a 'repair-prone' environment within the nucleus. Topologically associating domains are a key feature of 3D genome organization that compartmentalize transcription and replication, but little is known about their contribution to DNA repair processes2,3. Here we show that topologically associating domains are functional units of the DNA damage response, and are instrumental for the correct establishment of γH2AX-53BP1 chromatin domains in a manner that involves one-sided cohesin-mediated loop extrusion on both sides of the DSB. We propose a model in which H2AX-containing nucleosomes are rapidly phosphorylated as they actively pass by DSB-anchored cohesin. Our work highlights the importance of chromosome conformation in the maintenance of genome integrity and demonstrates the establishment of a chromatin modification by loop extrusion.

Mapping the signaling network of BIN2 kinase using TurboID-mediated biotin labeling and phosphoproteomics.

Elucidating enzyme-substrate relationships in posttranslational modification (PTM) networks is crucial for understanding signal transduction pathways but is technically difficult because enzyme-substrate interactions tend to be transient. Here, we demonstrate that TurboID-based proximity labeling (TbPL) effectively and specifically captures the substrates of kinases and phosphatases. TbPL-mass spectrometry (TbPL-MS) identified over 400 proximal proteins of Arabidopsis thaliana BRASSINOSTEROID-INSENSITIVE2 (BIN2), a member of the GLYCOGEN SYNTHASE KINASE 3 (GSK3) family that integrates signaling pathways controlling diverse developmental and acclimation processes. A large portion of the BIN2-proximal proteins showed BIN2-dependent phosphorylation in vivo or in vitro, suggesting that these are BIN2 substrates. Protein-protein interaction network analysis showed that the BIN2-proximal proteins include interactors of BIN2 substrates, revealing a high level of interactions among the BIN2-proximal proteins. Our proteomic analysis establishes the BIN2 signaling network and uncovers BIN2 functions in regulating key cellular processes such as transcription, RNA processing, translation initiation, vesicle trafficking, and cytoskeleton organization. We further discovered significant overlap between the GSK3 phosphorylome and the O-GlcNAcylome, suggesting an evolutionarily ancient relationship between GSK3 and the nutrient-sensing O-glycosylation pathway. Our work presents a powerful method for mapping PTM networks, a large dataset of GSK3 kinase substrates, and important insights into the signaling network that controls key cellular functions underlying plant growth and acclimation.

Repair of mismatched templates during Rad51-dependent Break-Induced Replication.

Using budding yeast, we have studied Rad51-dependent break-induced replication (BIR), where the invading 3' end of a site-specific double-strand break (DSB) and a donor template share 108 bp of homology that can be easily altered. BIR still occurs about 10% as often when every 6th base is mismatched as with a perfectly matched donor. Here we explore the tolerance of mismatches in more detail, by examining donor templates that each carry 10 mismatches, each with different spatial arrangements. Although 2 of the 6 arrangements we tested were nearly as efficient as the evenly-spaced reference, 4 were significantly less efficient. A donor with all 10 mismatches clustered at the 3' invading end of the DSB was not impaired compared to arrangements where mismatches were clustered at the 5' end. Our data suggest that the efficiency of strand invasion is principally dictated by thermodynamic considerations, i.e., by the total number of base pairs that can be formed; but mismatch position-specific effects are also important. We also addressed an apparent difference between in vitro and in vivo strand exchange assays, where in vitro studies had suggested that at a single contiguous stretch of 8 consecutive bases was needed to be paired for stable strand pairing, while in vivo assays using 108-bp substrates found significant recombination even when every 6th base was mismatched. Now, using substrates of either 90 or 108 nt-the latter being the size of the in vivo templates-we find that in vitro D-loop results are very similar to the in vivo results. However, there are still notable differences between in vivo and in vitro assays that are especially evident with unevenly-distributed mismatches. Mismatches in the donor template are incorporated into the BIR product in a strongly polar fashion up to ~40 nucleotides from the 3' end. Mismatch incorporation depends on the 3'→ 5' proofreading exonuclease activity of DNA polymerase δ, with little contribution from Msh2/Mlh1 mismatch repair proteins, or from Rad1-Rad10 flap nuclease or the Mph1 helicase. Surprisingly, the probability of a mismatch 27 nt from the 3' end being replaced by donor sequence was the same whether the preceding 26 nucleotides were mismatched every 6th base or fully homologous. These data suggest that DNA polymerase δ "chews back" the 3' end of the invading strand without any mismatch-dependent cues from the strand invasion structure. However, there appears to be an alternative way to incorporate a mismatch at the first base at the 3' end of the donor.

Higher Morbidity and Mortality in Women With Fournier Gangrene Compared With Men: Insights From National Inpatient Sample Data.

PURPOSE: To compare Fournier gangrene in female and male patients and identify mortality-associated characteristics in both. MATERIALS AND METHODS: We used National Inpatient Sample data (2016-2020) to identify Fournier gangrene cases and extracted demographic, comorbidity, and procedural variables. Multivariable regression models were used to identify mortality risk factors for both cohorts. RESULTS: We identified 2875 female (31%) and 6451 male patients (69%) with Fournier gangrene corresponding to an estimated 14,375 (95% CI, 13,784-14,966) and 32,255 (95% CI, 31,390-33,120) cases, respectively. Female patients were more likely to die than male patients (7.1% vs 5.7%, P < .0001, respectively). The median incidence rates were 1.7 (IQR, 1.5-1.8) and 4 (IQR, 3.6-4.3) cases per 100,000 person-years for female and male patients, respectively. Female patients had higher median age, longer hospital stays, more charges, procedures, and fecal diversion rates, but lower routine discharges than male patients (P < .05). Non-White female patients had increased mortality odds compared with White female patients (odds ratio [OR], 1.49; 95% CI, 1.07-2.07; P = .019). Prolonged interval until initial perineal debridement correlated with higher mortality odds in both female and male patients (OR, 1.02; 95% CI, 1-1.04; P = .034 vs OR, 1.03; 95% CI, 1.01-1.05; P < .0001). Diabetes lowered mortality odds in female and male patients (OR, 0.68; 95% CI, 0.47-0.99; P = .046 vs OR, 0.54; 95% CI, 0.41-0.7; P < .0001). CONCLUSIONS: In female patients, Fournier gangrene incidence surpasses previous reports, with slightly worse outcomes compared with male patients, emphasizing the need for precise clinical assessment and early intensive interventions.

The Impact of Social Deprivation on Anterior Urethral Stricture Recurrence After Urethroplasty - A Trauma and Urologic Reconstructive Network of Surgeons (TURNS) Analysis.

INTRODUCTION AND OBJECTIVES: Several factors influence recurrence after urethral stricture repair. The impact of socioeconomic factors on stricture recurrence after urethroplasty is poorly understood. This study aims to assess the impact that social deprivation, an area-level measure of disadvantage, has on urethral stricture recurrence after urethroplasty. METHODS: We performed a retrospective review of patients undergoing urethral reconstruction by surgeons participating in a collaborative research group. Home zip code was used to calculate Social Deprivation Indices (SDI; 0-100), which quantifies the level of disadvantage across several sociodemographic domains collected in the American Community Survey. Patients without zip code data were excluded from the analysis. The Cox Proportional Hazards model was used to study the association between SDI and the hazard of functional recurrence, adjusting for stricture characteristics as well as age and body mass index. RESULTS: Median age was 46.0 years with a median follow up of 367 days for the 1452 men included in the study. Patients in the fourth SDI quartile (worst social deprivation) were more likely to be active smokers with traumatic and infectious strictures compared to the first SDI quartile. Patients in the fourth SDI quartile had 1.64 times the unadjusted hazard of functional stricture recurrence vs patients in the first SDI quartile (95% CI 1.04-2.59). Compared to anastomotic ± excision, substitution only repair had 1.90 times the unadjusted hazard of recurrence. The adjusted hazard of recurrence was 1.08 per 10-point increase in SDI (95% CI 1.01-1.15, P = .027). CONCLUSIONS: Patient social deprivation identifies those at higher risk for functional recurrence after anterior urethral stricture repair, offering an opportunity for preoperative counseling and postoperative surveillance. Addressing these social determinants of health can potentially improve outcomes in reconstructive surgery.

Hydrated Formic Acid Clusters and their Interaction with Electrons.

We investigate ion formation in hydrated formic acid (FA) clusters upon collision with electrons of variable energy, focusing on electron ionization at 70 eV (EI) and low-energy (1.5-15 eV) electron attachment (EA). To uncover details about the composition of neutral clusters, we aim to elucidate the ion formation processes in FAM ⋅ WN clusters initiated by interaction with electrons and determine the extent of cluster fragmentation. EI predominantly produces protonated [FAm+H]+ ions, and in FA-rich clusters, the stable ring structures surrounding H3O+ ions are formed. In contrast, EA leads to a competition between the formation of intact [FAm ⋅ Wn]- and dissociated [FAm ⋅ Wn-H]- fragment ions, influenced by the cluster size, level of hydration, and electron energy. Our findings reveal a predisposition of low-energy EA towards forming [FAm ⋅ Wn]-, while higher electron energies tend to favor the formation of [FAm ⋅ Wn-H]- due to intracluster ion-molecule reactions. The comparison of positive and negative ion spectra suggests that the mass spectra of FA-rich clusters may indicate their actual size and composition. On the other hand, the more weakly bound water evaporation from the clusters depends strongly on the ionization. Thus, for the hydrated clusters, the neutral cluster size can hardly be estimated from the mass spectra.

Molecular characterization of Streptococcus suis isolates recovered from diseased pigs in Europe.

Streptococcus suis is a major swine pathogen and zoonotic agent, causing important economic losses to the porcine industry. Here, we used genomics approaches to characterize 251 S. suis isolates recovered from diseased pigs across Belgium, France, Germany, Hungary, the Netherlands, Spain, and the United Kingdom. We identified 13 serotypes, being serotypes 9 and 2 the most prevalent, and 34 sequence types (STs), including 16 novel STs, although ST16 and ST1 dominated the strain population. Phylogenetic analysis revealed complex genetic relationships, notable geographic clustering, and potential differential capacity for capsular switching among serotype 9 isolates. We found antimicrobial resistance (AMR) genes in 85.3% of the isolates, with high frequencies of genes conferring resistance to tetracyclines and macrolides. Specifically, 49.4% of the isolates harbored the tetO gene, and 64.9% possessed the ermB gene. Additionally, we observed a diverse array of virulence-associated genes (VAGs), including the classical VAGs mrp, epf, and sly, with variable presence across different genotypes. The high genetic diversity among European S. suis isolates highlights the importance of targeted antimicrobial use and flexible vaccine strategies. Rapid strain characterization is crucial for optimizing swine health management, enabling tailored interventions like the development of autovaccines to mitigate S. suis infections.

Fragility of overactive bladder medication clinical trials: A systematic review.

PURPOSE: Overactive bladder (OAB) syndrome significantly impairs quality of life, often necessitating pharmacological interventions with associated risks. The fragility of OAB trial outcomes, as measured by the fragility index (FI: smallest number of event changes to reverse statistical significance) and quotient (FQ: FI divided by total sample size expressed as a percentage), is critical yet unstudied. MATERIALS AND METHODS: We conducted a systematic search for randomized controlled trials on OAB medications published between January 2000 and August 2023. Inclusion criteria were trials with two parallel arms reporting binary outcomes related to OAB medications. We extracted trial details, outcomes, and statistical tests employed. We calculated FI and FQ, analyzing associations with trial characteristics through linear regression. RESULTS: We included 57 trials with a median sample size of 211 participants and a 12% median lost to follow-up. Most studies investigated anticholinergics (37/57, 65%). The median FI/FQ was 5/3.5%. Larger trials were less fragile (median FI 8; FQ 1.0%) compared to medium (FI: 4; FQ 2.5%) and small trials (FI: 4; FQ 8.3%). Double-blinded studies exhibited higher FQs (median 2.9%) than unblinded trials (6.7%). Primary and secondary outcomes had higher FIs (median 5 and 6, respectively) than adverse events (FI: 4). Each increase in 10 participants was associated with a +0.19 increase in FI (p < 0.001). CONCLUSIONS: A change in outcome for a median of five participants, or 3.5% of the total sample size, could reverse the direction of statistical significance in OAB trials. Studies with larger sample sizes and efficacy outcomes from blinded trials were less fragile.

Consumer product-related female genital injuries in the USA from 2013 to 2022.

BACKGROUND: Consumer product-related genital injuries in females across all age groups are understudied. Existing research focuses primarily on paediatric populations. We aimed to determine characteristics, trends and predictors of hospitalisation. METHODS: The National Electronic Injury Surveillance System database was queried for female genital injuries from 2013 to 2022. We stratified our population into four age groups (<18, 18-34, 35-54, >54 years). Automated text matching and manual reviews were employed for variable extraction. χ2 tests and logistic regression were conducted, accounting for survey design and weights. RESULTS: 9054 cases representing a national estimate of 252 329 injuries (95% CI 188 059 to 316 599) were identified. Paediatric injuries were most common (61%) and seniors had the highest hospitalisation rates (28%). Falls were common in paediatric (51%) and senior (48%) groups, whereas self-induced and topical application injuries were more frequent among adults aged 18-34 and 35-54. Injuries predominantly involved playground equipment and bicycles in children, razors and massage devices in adults aged 18-34 and 35-54 and household structures in seniors. Hospitalisation increased over the decade from 7% to 9%; significant predictors of hospitalisation were Asian race (OR=3.39, 95% CI 1.83 to 6.30), fractures (OR=7.98, 95% CI 4.85 to 13.1) and urethral injury (OR=3.15, 95% CI 1.30 to 7.63). CONCLUSIONS: Our study identifies distinct patterns in female genital injuries across ages. In the paediatric cohort, injuries are often linked to playgrounds and bicycles. For adults, grooming products are frequently implicated. Seniors commonly suffer injuries from household structures such as bathtubs. These patterns may inform discussions on tailored preventive strategies.

Comparing GPT-4 and Human Researchers in Health Care Data Analysis: Qualitative Description Study.

BACKGROUND: Large language models including GPT-4 (OpenAI) have opened new avenues in health care and qualitative research. Traditional qualitative methods are time-consuming and require expertise to capture nuance. Although large language models have demonstrated enhanced contextual understanding and inferencing compared with traditional natural language processing, their performance in qualitative analysis versus that of humans remains unexplored. OBJECTIVE: We evaluated the effectiveness of GPT-4 versus human researchers in qualitative analysis of interviews with patients with adult-acquired buried penis (AABP). METHODS: Qualitative data were obtained from semistructured interviews with 20 patients with AABP. Human analysis involved a structured 3-stage process-initial observations, line-by-line coding, and consensus discussions to refine themes. In contrast, artificial intelligence (AI) analysis with GPT-4 underwent two phases: (1) a naïve phase, where GPT-4 outputs were independently evaluated by a blinded reviewer to identify themes and subthemes and (2) a comparison phase, where AI-generated themes were compared with human-identified themes to assess agreement. We used a general qualitative description approach. RESULTS: The study population (N=20) comprised predominantly White (17/20, 85%), married (12/20, 60%), heterosexual (19/20, 95%) men, with a mean age of 58.8 years and BMI of 41.1 kg/m2. Human qualitative analysis identified "urinary issues" in 95% (19/20) and GPT-4 in 75% (15/20) of interviews, with the subtheme "spray or stream" noted in 60% (12/20) and 35% (7/20), respectively. "Sexual issues" were prominent (19/20, 95% humans vs 16/20, 80% GPT-4), although humans identified a wider range of subthemes, including "pain with sex or masturbation" (7/20, 35%) and "difficulty with sex or masturbation" (4/20, 20%). Both analyses similarly highlighted "mental health issues" (11/20, 55%, both), although humans coded "depression" more frequently (10/20, 50% humans vs 4/20, 20% GPT-4). Humans frequently cited "issues using public restrooms" (12/20, 60%) as impacting social life, whereas GPT-4 emphasized "struggles with romantic relationships" (9/20, 45%). "Hygiene issues" were consistently recognized (14/20, 70% humans vs 13/20, 65% GPT-4). Humans uniquely identified "contributing factors" as a theme in all interviews. There was moderate agreement between human and GPT-4 coding (κ=0.401). Reliability assessments of GPT-4's analyses showed consistent coding for themes including "body image struggles," "chronic pain" (10/10, 100%), and "depression" (9/10, 90%). Other themes like "motivation for surgery" and "weight challenges" were reliably coded (8/10, 80%), while less frequent themes were variably identified across multiple iterations. CONCLUSIONS: Large language models including GPT-4 can effectively identify key themes in analyzing qualitative health care data, showing moderate agreement with human analysis. While human analysis provided a richer diversity of subthemes, the consistency of AI suggests its use as a complementary tool in qualitative research. With AI rapidly advancing, future studies should iterate analyses and circumvent token limitations by segmenting data, furthering the breadth and depth of large language model-driven qualitative analyses.

Negative Nonlinear CD-ee Dependence in Polycrystalline BINOL Thin Films.

Generally, the relationship between the observed circular dichroism and the enantiomeric excess in chiral systems (CD-ee dependence) is linear. While positive nonlinear behavior has often been reported in the past, examples of negative nonlinear (NN) behavior in CD-ee dependence are rare and not well understood. Here, we present a strong NN CD-ee dependence within polycrystalline thin films of BINOL by using second-harmonic-generation circular dichroism (SHG-CD) and commercial CD spectroscopy studies. Theoretical calculations, microscopy, and FTIR studies are employed to further clarify the underlying cause of this observation. This behavior is attributed to the changing supramolecular chirality of the system. Systems exhibiting NN CD-ee dependence hold promise for highly accurate enantiomeric excess characterization, which is essential for the refinement of enantio-separating and -purifying processes in pharmaceuticals, asymmetric catalysis, and chiral sensing. Our findings suggest that a whole class of single-species systems, i.e., racemate crystals, might possess NN CD-ee dependence and thus provide us a vast playground to better understand and exploit this phenomenon.

Pancreatic Cyst Surveillance: Who, Why, How?

Pancreatic cystic lesions (PCLs) are widely prevalent and commonly encountered in abdominal radiology. Some PCLs can be definitively identified at imaging as benign subtypes or those with malignant potential, while others remain indeterminate. Notably, the degree of malignant potential and natural history of the most common subtype, branch-duct intraductal papillary mucinous neoplasms, are not clearly established. In the work-up of PCLs, patients may further be identified as high-risk individuals who are at elevated risk of pancreatic ductal adenocarcinoma due to familial and genetic factors. This review describes current PCL surveillance and management guidelines and highlights ongoing controversies and future directions to aid radiologists in their daily practice.

Haemorrhagic cystitis: a review of management strategies and emerging treatments.

Haemorrhagic cystitis (HC) is characterised by persistent haematuria and lower urinary tract symptoms following radiotherapy or chemotherapy. Its pathogenesis is poorly understood but thought to be related to acrolein toxicity following chemotherapy or fibrosis/vascular remodelling after radiotherapy. There is no standard of care for patients with HC, although existing strategies including fulguration, hyperbaric oxygen therapy, botulinum toxin A, and other intravesical therapies have demonstrated short-term efficacy in cohort studies. Novel agents including liposomal tacrolimus are promising targets for further research. This review summarises the incidence and pathogenesis of HC as well as current evidence supporting its different management strategies.

Tree identity and canopy openness mediate oil palm biodiversity enrichment effects on insect herbivory and pollination.

As the extent of oil palm (Elaeis guineensis) cultivation has expanded at the expense of tropical rainforests, enriching conventional large-scale oil palm plantations with native trees has been proposed as a strategy for restoring biodiversity and ecosystem function. However, how tree enrichment affects insect-mediated ecosystem functions is unknown. We investigated impacts on insect herbivory and pollination in the fourth year of a plantation-scale, long-term oil palm biodiversity enrichment experiment in Jambi, Sumatra, Indonesia. Within 48 plots systematically varying in size (25-1600 m2 ) and planted tree species richness (one to six species), we collected response data on vegetation structure, understory insect abundances, and pollinator and herbivore activity on chili plants (Capsicum annuum), which served as indicators of insect-mediated ecosystem functions. We examined the independent effects of plot size, tree species richness, and tree identity on these response variables, using the linear model for random partitions design. The experimental treatments were most associated with vegetation structure: tree identity mattered, as the species Peronema canescens strongly decreased (by approximately one standard deviation) both canopy openness and understory vegetation cover; whereas tree richness only decreased understory flower density. Further, the smallest plots had the lowest understory flower density and richness, presumably because of lower light availability and colonization rates, respectively. Enrichment influenced herbivorous insects and natural enemies in the understory to a lesser extent: both groups had higher abundances in plots with two enrichment species planted, possibly because higher associated tree mortality created more habitat, while herbivores decreased with increasing tree species richness, in line with the resource concentration hypothesis. Linking relationships in structural equation models showed that the negative association between P. canescens and understory vegetation cover was mediated through canopy openness. Likewise, canopy openness mediated increases in herbivore and pollinator insect abundances. Higher pollinator visitation increased phytometer yield, while impacts of insect herbivores on yield were not apparent. Our results demonstrate that even at an early stage, different levels of ecological restoration influence insect-mediated ecosystem functions, mainly through canopy openness. These findings suggest that maintaining some canopy gaps while enrichment plots develop may be beneficial for increasing habitat heterogeneity and insect-mediated ecosystem functions.

Novel series of tunable µOR modulators with enhanced brain penetration for the treatment of opioid use disorder, pain and neuropsychiatric indications.

Structural optimization of a previously reported agonist of µOR, PZM21 is described resulting in the discovery of a novel series of amides with at least 4-folds enhanced CNS penetration in rat. Furthermore, these efforts yielded compounds with varying levels of efficacy on the receptor ranging from high efficacy agonists such as compound 20 to antagonists, such as 24. The correlation between in vitro activation of µOR and relative activity in models of analgesia for these compounds is discussed. The compelling results obtained in these studies demonstrate the potential utility of these newly discovered compounds in the treatment of pain and opioid use disorder.

Rad51-mediated double-strand break repair and mismatch correction of divergent substrates.

The Rad51 (also known as RecA) family of recombinases executes the critical step in homologous recombination: the search for homologous DNA to serve as a template during the repair of DNA double-strand breaks (DSBs). Although budding yeast Rad51 has been extensively characterized in vitro, the stringency of its search and sensitivity to mismatched sequences in vivo remain poorly defined. Here, in Saccharomyces cerevisiae, we analysed Rad51-dependent break-induced replication in which the invading DSB end and its donor template share a 108-base-pair homology region and the donor carries different densities of single-base-pair mismatches. With every eighth base pair mismatched, repair was about 14% of that of completely homologous sequences. With every sixth base pair mismatched, repair was still more than 5%. Thus, completing break-induced replication in vivo overcomes the apparent requirement for at least 6-8 consecutive paired bases that has been inferred from in vitro studies. When recombination occurs without a protruding nonhomologous 3' tail, the mismatch repair protein Msh2 does not discourage homeologous recombination. However, when the DSB end contains a 3' protruding nonhomologous tail, Msh2 promotes the rejection of mismatched substrates. Mismatch correction of strand invasion heteroduplex DNA is strongly polar, favouring correction close to the DSB end. Nearly all mismatch correction depends on the proofreading activity of DNA polymerase-δ, although the repair proteins Msh2, Mlh1 and Exo1 influence the extent of correction.