Selective permeation up a chemical potential gradient to enable an unusual solvent purification modality.

The need for energy-efficient recovery of organic solutes from aqueous streams is becoming more urgent as chemical manufacturing transitions toward nonconventional and bio-based feedstocks and processes. In addition to this, many aqueous waste streams contain recalcitrant organic contaminants, such as pharmaceuticals, industrial solvents, and personal care products, that must be removed prior to reuse. We observe that rigid carbon membrane materials can remove and concentrate organic contaminants via an unusual liquid-phase membrane permeation modality. Surprisingly, detailed thermodynamic calculations on the chemical potential of the organic contaminant reveal that the organic species has a higher chemical potential on the permeate side of the membrane than on the feed side of the membrane. This unusual observation challenges conventional membrane transport theory that posits that all permeating species move from high chemical potential states to lower chemical potential states. Based on experimental measurements, we hypothesize that the organic is concentrated in the membrane relative to water via favorable binding interactions between the organic and the carbon membrane. The concentrated organic is then swept through the membrane via the bulk flow of water in a modality known as "sorp-vection." We highlight via simplified nonequilibrium thermodynamic models that this "uphill" chemical potential permeation of the organic does not result in second-law violations and can be deduced via measurements of the organic and water sorption and diffusion rates into the carbon membrane. Moreover, this work identifies the need to consider such nonidealities when incorporating unique, rigid materials for the separations of aqueous waste streams.

The SMX DNA Repair Tri-nuclease.

The efficient removal of replication and recombination intermediates is essential for the maintenance of genome stability. Resolution of these potentially toxic structures requires the MUS81-EME1 endonuclease, which is activated at prometaphase by formation of the SMX tri-nuclease containing three DNA repair structure-selective endonucleases: SLX1-SLX4, MUS81-EME1, and XPF-ERCC1. Here we show that SMX tri-nuclease is more active than the three individual nucleases, efficiently cleaving replication forks and recombination intermediates. Within SMX, SLX4 co-ordinates the SLX1 and MUS81-EME1 nucleases for Holliday junction resolution, in a reaction stimulated by XPF-ERCC1. SMX formation activates MUS81-EME1 for replication fork and flap structure cleavage by relaxing substrate specificity. Activation involves MUS81's conserved N-terminal HhH domain, which mediates incision site selection and SLX4 binding. Cell cycle-dependent formation and activation of this tri-nuclease complex provides a unique mechanism by which cells ensure chromosome segregation and preserve genome integrity.

A propensity score matched analysis of liver transplantation outcomes in the setting of preservation solution shortage.

The recent shortage of the University of Wisconsin (UW) solution prompted increased utilization of histidine-tryptophan-ketoglutarate (HTK) solution for liver graft preservation. This contemporary study analyzed deceased donor liver transplant outcomes following preservation with HTK vs UW. Patients receiving deceased donor liver transplantations between January 1, 2019, and June 30, 2022, were retrospectively identified utilizing the Organ Procurement and Transplant Network database, stratified by preservation with HTK vs UW, and a propensity score matching analysis was performed. Outcomes assessed included rates of primary nonfunction, graft survival, and patient survival. There were 4447 patients in each cohort. Primary nonfunction occurred in 60 (1.35%) patients in the HTK group vs 25 (0.54%) in the UW group (P < .001). HTK was associated with lower 90-day graft survival (94.39% vs 96.09%; P < .001) and 90-day patient survival (95.97% vs 97.38%; P = .001). Unmatched donation after cardiac death-specific analysis of HTK vs UW demonstrated respective rates of primary nonfunction of 1.63% vs 0.82% (P = .20), 90-day graft survival of 92.50% vs 95.29% (P = .069), and 90-day patient survival of 93.90% vs 96.35% (P = .077). These results suggest that HTK may not be an equivalent preservation solution for deceased donor liver transplantation.

Reversible Chemical Modification of Antibody Effector Function Mitigates Unwanted Systemic Immune Activation.

Antibody effector functions including antibody-dependent cellular cytotoxicity (ADCC) and phagocytosis (ADCP) are mediated through the interaction of the antibody Fc region with Fcγ receptors present on immune cells. Several approaches have been used to modulate antibody Fc-Fcγ interactions with the goal of driving an effective antitumor immune response, including Fc point mutations and glycan modifications. However, robust antibody-Fcγ engagement and immune cell binding of Fc-enhanced antibodies in the periphery can lead to the unwanted induction of systemic cytokine release and other dose-limiting infusion-related reactions. Creating a balance between effective engagement of Fcγ receptors that can induce antitumor activity without incurring systemic immune activation is an ongoing challenge in the field of antibody and immuno-oncology therapeutics. Herein, we describe a method for the reversible chemical modulation of antibody-Fcγ interactions using simple poly(ethylene glycol) (PEG) linkers conjugated to antibody interchain disulfides with maleimide attachments. This method enables dosing of a therapeutic with muted Fcγ engagement that is restored in vivo in a time-dependent manner. The technology was applied to an effector function enhanced agonist CD40 antibody, SEA-CD40, and experiments demonstrate significant reductions in Fc-induced immune activation in vitro and in mice and nonhuman primates despite showing retained efficacy and improved pharmacokinetics compared to the parent antibody. We foresee that this simple, modular system can be rapidly applied to antibodies that suffer from systemic immune activation due to peripheral FcγR binding immediately upon infusion.

A systematic review of focused ultrasound for psychiatric disorders: current applications, opportunities, and challenges.

OBJECTIVE: Advancements in MRI-guided focused ultrasound (MRgFUS) technology have led to the successful treatment of select movement disorders. Based on the comparative success between ablation and deep brain stimulation, interest arises in focused ultrasound (FUS) as a promising treatment modality for psychiatric illnesses. In this systematic review, the authors examined current applications of FUS for psychiatric conditions and explored its potential opportunities and challenges. METHODS: The authors performed a comprehensive review using the PRISMA guidelines of studies investigating psychiatric applications for FUS. Articles indexed on PubMed between 2014 to 2024 were included. The authors synthesized the psychiatric conditions treated, neural targets, outcomes, study design, and sonication parameters, and they reviewed important considerations for the treatment of psychiatric disorders with FUS. They also discussed active clinical trials in this research domain. RESULTS: Of 250 articles, 10 met the inclusion criteria. Eight articles investigated the clinical, safety, and imaging correlates of MRgFUS in obsessive-compulsive disorder (OCD), whereas 3 examined treatment-resistant depression. Bilateral anterior capsulotomy resulted in a full responder rate of 67% (≥ 35% reduction in the Yale-Brown Obsessive-Compulsive Scale score) and 33% (≥ 50% reduction in the score on the Hamilton Rating Scale for Depression) in OCD and treatment-resistant depression, respectively. Sonications ranged from 8 to 36 with targeted lesional temperatures of 51°C-56°C. Lesions in the anterodorsal aspect of the anterior limb of the internal capsule (ALIC) and increased functional connectivity to the left dorsolateral prefrontal cortex and dorsal anterior cingulate cortex significantly predicted reduction in symptoms among patients with OCD, with decreases in beta-band activity in the frontocentral and temporal regions associated with reductions in depression and anxiety. Treatment of the nucleus accumbens with low-intensity FUS (LIFU) in patients with opioid-use disorders resulted in significant reductions in cue-reactive cravings, lasting up to 90 days. No serious adverse events were reported, including cognitive decline. Side effects were generally mild and transient, consisting of headaches, pin-site swelling, and nausea. Fourteen active clinical trials were identified, primarily targeting depression with LIFU. CONCLUSIONS: Currently, FUS for psychiatric conditions is centered on OCD, with early pilot studies demonstrating promising safety and efficacy. Further research expanding on defining optimal patient selection, study design, intensity, and sonication parameters is warranted, particularly as FUS expands to other psychiatric illnesses and incorporates LIFU paradigms. Ethical considerations such as patient consent and equitable access also remain paramount.

Attitudes and professional values of veterinarians and veterinary students toward positive welfare states for dairy cattle.

Research that involves agricultural animal welfare has typically aimed to improve welfare by decreasing disease, distress, and pain. Positive welfare does not necessarily occur with the absence of suffering but in combination with opportunities for behaviors or affective states desired by animals. Our objectives were to describe Canadian bovine veterinarians' and veterinary students' attitudes, professional normative values, and perceived ability to promote positive welfare for dairy cows, and to explore participants' provided rationale. With an online cross-sectional survey, Canadian veterinary practitioners (n = 78) and veterinary students (n = 148) were asked, on a 7-point Likert scale, about their attitudes, perceived professional normative values, and perceived ability of veterinarians to promote positive welfare for dairy cows. We used an applied thematic analysis approach within the qualitative description methodology to analyze participants' open-ended text responses. Quantitatively, participants had very favorable attitudes (mean ± SE; 6.3 ± 0.04) and perceived favorable values (5.7 ± 0.06) in the veterinary community toward positive welfare opportunities for dairy cows. Three themes were identified to explain the professional normative values: influences from within the veterinary profession, influences from outside the veterinary profession, and personal views of participants. Participants were confident that veterinarians could suggest positive welfare opportunities for dairy cows (6.1 ± 0.06) but were uncertain that the decision to suggest these opportunities to producers was within a veterinarian's control (4.3 ± 0.11) and were not confident that implementation of positive welfare opportunities was under a veterinarian's control (2.1 ± 0.07). Three themes were identified to explain the barriers to veterinarians promoting positive welfare opportunities for dairy cows: not practical to implement, resistance to change, and concern for the animal. Participants stated that many positive welfare opportunities were impractical or expensive to implement. We conclude that positive attitudes and positive professional values exist in the veterinary community toward positive welfare for dairy cows but that much uncertainty exists regarding a veterinarian's ability to influence change to current practices.

Can 60 days of feeding lead to increased fitness for transport in cull dairy cows?

The welfare of cull cows during transport to slaughter is a current concern in the Canadian dairy industry. Cull cows sold through auction often have a high prevalence of lameness, low BCS, hock lesions, and udder engorgement. To evaluate whether drying off and feeding cull dairy cows before transport can mitigate these challenges, 45 cows designated for culling were randomly assigned to either be fed for 60 d after being dried off (fed group; n = 24) or to serve as controls by being sent directly to slaughter (direct group; n = 21). Two fed group cows were removed for health reasons before completing the feeding period. Both fed group and direct group cows were assessed for locomotion (5-point scale), BCS (5-point scale), hock lesions (3-point scale), udder engorgement (3-point scale), and BW at the time of enrollment. Fed group cows, locomotion, BCS, hock, and udder engorgement scores were assessed weekly until slaughter. Weights of the fed group cows were measured again the day before slaughter. Mixed linear regression models were used to assess continuous outcomes BCS and weight. Mixed logistic regression models were used to assess dichotomous outcomes presence of hock lesions and lameness. Fed group cows gained an average of 116.9 kg over the feeding period (SE ± 8.20). Fed group cows had an average weight at slaughter of 834.2 kg, whereas direct group cows' average weight was 767.3 kg (SE ± 26.8). The fed group cows' average BCS at the start of the trial was 2.4, and at slaughter was 3.6, with an average gain of 1.2 BCS points. At slaughter, the proportion of udders involuted in the fed group was 45.1% (10/22) and in the direct group cows, was 0% (0/21). No differences were found in locomotion or hock lesions between the fed group and direct groups. It is important to weigh potential benefits for the fed group cows with the fact that direct group cows did not endure a drying off procedure, nor were they placed at risk of potential adverse health events. However, despite these potential limitations, due to the improved BCS and udder engorgement scores, cows fed for 60 d may be better prepared for the transportation to slaughter, as well as sell for a higher price due to increased BW and body condition.

Improving market (cull) dairy cows carcass traits and meat quality.

Despite a high volume of market (cull) dairy cows entering the food chain every year, beef from market dairy cows is largely considered insignificant when compared with beef from beef cattle in the market and is widely thought to be used for ground beef only. Our study aimed to evaluate the effects of feeding dairy market cows on a lactating TMR diet before slaughter on carcass and beef quality traits. Forty-three Holstein market cows were randomly assigned into 2 treatments: Fed or Direct. Fed cows (n = 22) were dried off, then fed a lactating cow TMR ration for approximately 60 d whereas Direct cows (n = 21) were sent directly to slaughter. Hot carcass weight (HCW) was used to calculate the dressing percentage (DP) together with the animals' body weight recorded when animals left the farm to go to the abattoir. At 24h post-mortem, the rib fat thickness (RFT) and rib eye area (REA) were measured by a certified grader. Afterward, rib samples were collected between the 12th -10th ribs from one side of the carcass and divided into 5 ribeye steaks. The first steak was used for intramuscular fat (IMF) content analysis by NIRS. The other 4 steaks were then aged for 7, 14, 21, or 28 d. Following aging, the steaks available were assessed for tenderness using the Warner-Bratzler Shear Force protocol. Data were analyzed by mixed linear regression to compare experimental groups for continuous outcomes. HCW of the Fed cows was higher than the Direct cows, with a mean HCW of 408.3 kg and 326.1 kg, respectively (SE ± 9.0). Average DP in Fed cows was 49.1%, compared with 42.5% in Direct cows (SE ± 0.69). The average percentage of IMF in Fed cows was 8.1% and in Direct cows was 5.0% (SE ± 0.43). Fed cows had a mean REA of 68.1 cm2 and Direct cows had a mean of 57.7 cm2 (SE ± 11.9). No differences were found for RFT. The steaks aged for 14 d resulted in meat with a mean shear force (SF) of 4.19 kg in Fed cows, while Direct cows had a mean SF of 6.28 kg (SE ± 0.29) Feeding market dairy cows for 60 d before slaughter enhanced carcass weight and yield, IMF content, and tenderness. These results indicate that approximately 60 d of feeding can improve the quality of market dairy cow meat and may help improve market cow value.

Effect of training method and dairy cow personality traits on adaptation to an automated milking system.

Training dairy cows on the automated milking system (AMS) can improve their performance, however, the ideal training method and how their individual personality relates to training is unknown. The objectives of this study were to determine: 1) the effect of AMS training method on adaptation of primiparous cows to an AMS, 2) whether the personality of a cow affects their adaptation to the AMS, and 3) whether certain personality types adapt better based on the type of training they receive. Forty-eight primiparous Holstein dairy cows, who had not previously been milked by an AMS, were enrolled in the study at ∼85 DIM and were assessed for personality traits using a combined arena test comprised of 3 parts: a novel arena test, a novel object test, and a novel human test. Cows were allocated into 1 of 4 AMS training treatments (n = 12): control (CON; no training), environment exposure only (ENV), environment and AMS concentrate provision (PEL), and environment and mechanics exposure (MEC). Cows in the ENV, PEL, and MEC treatments were subjected to training 4 × /d for 4 d, and then cows from all 4 treatments were milked on the AMS in the evening of the 4th training d. Milking activity and production were recorded for the first 14 d on the AMS. PEL cows had fewer visits to the AMS, fewer milkings, and fewer voluntary milkings compared with CON cows across the 14 d. Cows on all experimental treatments had more fetching events than CON cows, while MEC and ENV treatment cows had a higher milk yield compared with those on the CON treatment. Principal component analysis of the behaviors recorded during the personality assessment revealed 2 factors interpreted as personality traits, "explore-active" and "bold." Across all cows, more "explore-active" cows had greater milk yield. Only the performance of ENV cows was associated with personality; ENV cows who scored low for "explore-active" differed from CON cows, through increased visits and voluntary milkings. Overall, the results demonstrate that providing concentrate on its own during AMS training may not be motivating enough to improve adaptation and performance. However, exposing cows to the mechanics (e.g., the robotic arm and sounds of the AMS) before first milking may improve milk yield. Finally, cow personality may influence the performance of cows trained using less complicated training methods, such as only exposing them to the environment of the AMS.

Similar two-year improvement in patient-reported outcomes in patients 40 years and older with outerbridge grade III and IV lesions following ACL reconstruction.

PURPOSE: This study aims to evaluate clinical and patient-reported outcome measures (PROMs) of patients 40 years and older that underwent anterior cruciate ligament reconstruction (ACLR) and determine the influence of preexisting arthritis and chondral wear on ACLR outcomes. We hypothesized that patients aged 40+ with ACLR would have excellent clinical outcomes and PROMs regardless of preexisting arthritic changes. METHODS: A total of 118 patients were included. Patients aged 40 years and older who underwent ACLR in a single healthcare system between 2009 and 2016 were eligible. Outcomes assessed include Knee Injury and Osteoarthritis Outcome Scores (KOOS), Single Assessment Numeric Evaluation (SANE) scores, intraoperative Outerbridge grading, preoperative Kellgren-Lawrence (KL) grading and postoperative complication rates over a 2-year period. RESULTS: Average increase in KOOS and SANE scores were 21.2 ± $\pm $ 19.9 and 23.5 ± $\pm $ 31.3, respectively. Patients with Outerbridge grade III and IV lesions in weight-bearing compartments had lower baseline SANE and KOOS scores than those without (47.1 ± 22.0 vs. 64.5 ± 23.6 baseline SANE and 43.1 ± 18.1 vs. 63.5 ± 15.9 baseline KOOS; p = 0.002 and p < 0.001, respectively) with no significant difference in the amount of change in SANE or KOOS scores (p = 0.111 and p = 0.165 respectively). Patients with KL-grade 2+ osteoarthritis experienced similar changes in KOOS and SANE over the 2-year period to their counterparts (p = 0.598 and p = 0.643, respectively). CONCLUSION: There is no correlation between preexisting osteoarthritic changes or chondral defects and PROs. KOOS and SANE scores both increased postoperatively. When treating older patients with an ACL tear, surgeons should consider the activity level and desires of the patient as they determine appropriate treatment. Preexisting osteoarthritis does not correlate with patient-reported outcomes for ACLR. LEVEL OF EVIDENCE: Level IV.

An open trial for adults in a residential program for binge eating spectrum disorders.

The current study assessed treatment outcome for 99 adult admissions to a residential program specifically designed for binge eating spectrum disorders (BESD). Participants completed self-report measures at admission, discharge, and 12-month follow-up and were asked to complete blood draws at admission and follow-up. Primary outcomes were eating behaviors; secondary outcomes included comorbid symptoms and physiological variables. Eating behaviors improved significantly from admission to follow-up, as did most comorbid symptoms and quality of life, despite no change in body mass index. Some variables displayed a curvilinear relationship, with some worsening of symptoms from discharge to follow-up, although scores at follow-up remained well below admission values. Participation in the treatment program was associated with reduced problematic eating and comorbid symptoms and increased quality of life up to one year after discharge. Findings from this study may encourage the development of similar residential treatment programs for BESD for those who have not responded to outpatient care or mixed milieu settings, and may prompt randomized studies testing similar treatments versus usual care.

Comprehensive Interactome Mapping of the DNA Repair Scaffold SLX4 Using Proximity Labeling and Affinity Purification.

The DNA repair scaffold SLX4 has pivotal roles in cellular processes that maintain genome stability, most notably homologous recombination. Germline mutations in SLX4 are associated with Fanconi anemia, a disease characterized by chromosome instability and cancer susceptibility. The role of mammalian SLX4 in homologous recombination depends critically on binding and activating structure-selective endonucleases, namely SLX1, MUS81-EME1, and XPF-ERCC1. Increasing evidence indicates that cells rely on distinct SLX4-dependent complexes to remove DNA lesions in specific regions of the genome. Despite our understanding of SLX4 as a scaffold for DNA repair proteins, a detailed repertoire of SLX4 interactors has never been reported. Here, we provide a comprehensive map of the human SLX4 interactome using proximity-dependent biotin identification (BioID) and affinity purification coupled to mass spectrometry (AP-MS). We identified 221 unique high-confidence interactors, of which the vast majority represent novel SLX4-binding proteins. Network analysis of these hits revealed pathways with known involvement of SLX4, such as DNA repair, and several emerging pathways of interest, including RNA metabolism and chromatin remodeling. In summary, the comprehensive SLX4 interactome we report here provides a deeper understanding of how SLX4 functions in DNA repair while revealing new cellular processes that may involve SLX4.

Prevalence and Characteristics of Diagnostic Error in Pediatric Critical Care: A Multicenter Study.

OBJECTIVES: Effective interventions to prevent diagnostic error among critically ill children should be informed by diagnostic error prevalence and etiologies. We aimed to determine the prevalence and characteristics of diagnostic errors and identify factors associated with error in patients admitted to the PICU. DESIGN: Multicenter retrospective cohort study using structured medical record review by trained clinicians using the Revised Safer Dx instrument to identify diagnostic error (defined as missed opportunities in diagnosis). Cases with potential errors were further reviewed by four pediatric intensivists who made final consensus determinations of diagnostic error occurrence. Demographic, clinical, clinician, and encounter data were also collected. SETTING: Four academic tertiary-referral PICUs. PATIENTS: Eight hundred eighty-two randomly selected patients 0-18 years old who were nonelectively admitted to participating PICUs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Of 882 patient admissions, 13 (1.5%) had a diagnostic error up to 7 days after PICU admission. Infections (46%) and respiratory conditions (23%) were the most common missed diagnoses. One diagnostic error caused harm with a prolonged hospital stay. Common missed diagnostic opportunities included failure to consider the diagnosis despite a suggestive history (69%) and failure to broaden diagnostic testing (69%). Unadjusted analysis identified more diagnostic errors in patients with atypical presentations (23.1% vs 3.6%, p = 0.011), neurologic chief complaints (46.2% vs 18.8%, p = 0.024), admitting intensivists greater than or equal to 45 years old (92.3% vs 65.1%, p = 0.042), admitting intensivists with more service weeks/year (mean 12.8 vs 10.9 wk, p = 0.031), and diagnostic uncertainty on admission (77% vs 25.1%, p < 0.001). Generalized linear mixed models determined that atypical presentation (odds ratio [OR] 4.58; 95% CI, 0.94-17.1) and diagnostic uncertainty on admission (OR 9.67; 95% CI, 2.86-44.0) were significantly associated with diagnostic error. CONCLUSIONS: Among critically ill children, 1.5% had a diagnostic error up to 7 days after PICU admission. Diagnostic errors were associated with atypical presentations and diagnostic uncertainty on admission, suggesting possible targets for intervention.

HPV vaccination uptake among foreign-born Blacks in the US: insights from the National Health Interview Survey 2013-2017.

INTRODUCTION: Human papillomavirus (HPV) vaccination uptake is lower among foreign-born than US-born individuals, but HPV-related (e.g., cervical) cancer risks are disproportionately higher among immigrant populations. Although timely vaccination can help reduce these risks, less is known about differences in the low HPV vaccination uptake among foreign-born groups, especially Black immigrants. The purpose of this study was to examine the differences in HPV vaccination initiation among US- and foreign-born Black men and women. METHOD: Data from the 2013-2017 National Health Interview Survey on Black adults, aged 18-37 years, were analyzed in 2019. HPV vaccination initiation prevalence among US- and foreign-born blacks by region of birth were examined. Multivariate binary logistic regression analysis was used to examine the relationship between foreign-birth status and HPV vaccination initiation separately among men and women, after adjusting for sociodemographic and health-related factors. RESULTS: There were significant differences (p < 0.001) in HPV vaccination initiation among Blacks from the US (22.5%), Africa (14.2%), and Americas/Caribbean Islands (11.4%). Adjusted odds of HPV vaccination initiation were lower among foreign- than US-born Blacks (AOR 0.71, CI 0.52, 0.98) but insignificant after controlling for health-related factors. Being ≤ 17 years versus 18-26 years at age of vaccine eligibility (AOR 3.44, CI 2.90, 4.07) was associated with HPV vaccination, and this relationship remained significant among men and women. Being single was associated with vaccination initiation among men, and some college experience, fair/poor health, obstetric/gynecological visit, and pap test were associated with HPV vaccination. Conclusion Cancer prevention strategies to promote HPV vaccination should consider making age-appropriate, gender-specific, and culturally relevant programs among foreign-born blacks in the US. Health insurance is also a key factor that might help with the lower rates of vaccinated black immigrants.

Development of a yeast whole-cell biocatalyst for MHET conversion into terephthalic acid and ethylene glycol.

BACKGROUND: Over the 70 years since the introduction of plastic into everyday items, plastic waste has become an increasing problem. With over 360 million tonnes of plastics produced every year, solutions for plastic recycling and plastic waste reduction are sorely needed. Recently, multiple enzymes capable of degrading PET (polyethylene terephthalate) plastic have been identified and engineered. In particular, the enzymes PETase and MHETase from Ideonella sakaiensis depolymerize PET into the two building blocks used for its synthesis, ethylene glycol (EG) and terephthalic acid (TPA). Importantly, EG and TPA can be re-used for PET synthesis allowing complete and sustainable PET recycling. RESULTS: In this study we used Saccharomyces cerevisiae, a species utilized widely in bioindustrial fermentation processes, as a platform to develop a whole-cell catalyst expressing the MHETase enzyme, which converts monohydroxyethyl terephthalate (MHET) into TPA and EG. We assessed six expression architectures and identified those resulting in efficient MHETase expression on the yeast cell surface. We show that the MHETase whole-cell catalyst has activity comparable to recombinant MHETase purified from Escherichia coli. Finally, we demonstrate that surface displayed MHETase is active across a range of pHs, temperatures, and for at least 12 days at room temperature. CONCLUSIONS: We demonstrate the feasibility of using S. cerevisiae as a platform for the expression and surface display of PET degrading enzymes and predict that the whole-cell catalyst will be a viable alternative to protein purification-based approaches for plastic degradation.

The Society for Vascular Surgery Objective Performance Goals for Critical Limb Ischemia are attainable in select patients with ischemic wounds managed with wound care alone.

BACKGROUND: To set therapeutic benchmarks, in 2009 the Society for Vascular Surgery defined objective performance goals (OPG) for treatment of patients with chronic limb threatening ischemia (CLTI) with either open surgical bypass or endovascular intervention. The goal of these OPGs are to set standards of care from a revascularization standpoint and to provide performance benchmarks for 1 year patency rates for new endovascular therapies. While OPGs are useful in this regard, a critical decision point in the treatment of patients with CLTI is determining when revascularization is necessary. There is little guidance in the comprehensive treatment of this patient population, especially in the nonoperative cohort. Guidelines are needed for the CLTI patient population as a whole and not just those revascularized, and our aim was to assess whether CLTI OPGs could be attained with nonoperative management alone. METHODS: Our cohort included patients with an incident diagnosis of CLTI (by hemodynamic and symptomatic criteria) at our institution from 2013-2017. The primary outcome measured was mortality. Secondary outcomes were limb loss and failure of amputation-free survival. Descriptive statistics were used to define the 2 groups - patients undergoing primary revascularization and patients undergoing primary wound management. The risk difference in outcomes between the 2 groups was estimated using collaborative-targeted maximum likelihood estimation. RESULTS: Our cohort included 349 incident CLTI patients; 60% male, 51% white, mean age 63 +/- 13 years, 20% Rutherford 4, and 80% Rutherford 5. Most patients (277, 79%) underwent primary revascularization, and 72 (21%) were treated with wound care alone. Demographics and presenting characteristics were similar between groups. Although the revascularized patients were more likely to have femoropopliteal disease (72% vs. 36%), both groups had a high rate of infrapopliteal disease (62% vs. 57%). Not surprisingly, the patients in the revascularization group were less likely to have congestive heart failure (34% vs. 42%), complicated diabetes (52% vs. 79%), obesity (19% vs. 33%), and end stage renal disease (14% vs. 28%). In the wound care group, 2-year outcomes were 65% survival, 51% amputation free survival, 19% major limb amputation, and 17% major adverse cardiac event. The wound care cohort had a 13% greater risk of death at 2 years; however, the risk of limb loss at 2 years was 12% less in the wound care cohort. CONCLUSIONS: A comprehensive set treatment goals and expected amputation free survival outcomes can guide revascularization, but also assure that appropriate outcomes are achieved for patients treated without revascularization. The 2-year outcomes achieved in this cohort provide an estimate of outcomes for nonrevascularized CLTI patients. Although multi-center or prospective studies are needed, we demonstrate that equal, even improved, limb salvage rates are possible.

Predictors of diarrhea, mortality, and weight gain in male dairy calves.

The objective of this prospective cohort study was to determine the effect of an abnormal fecal consistency score on weight gain and mortality in male Holstein calves and to identify risk factors associated with the occurrence of an abnormal fecal consistency score. This study enrolled 2,616 calves entering a calf-raising facility in Ontario, Canada, between January 2018 and December 2020. Fecal consistency scores were assigned twice daily for the first 28 d following arrival, where a score of 2, indicating runny consistency, and 3, indicating watery consistency, were classified as diarrhea. Severe diarrhea was classified by a score of 3. Serum total protein was measured upon arrival and the source of the calf (i.e., whether the calf came from a drover, local farm, or auction) was recorded. Body weight measurements were also collected at arrival and at 14, 56, and 77 d after arrival. Calf mortality and disease treatment during the first 77 d were recorded. On average, calves had diarrhea for 16% (4.51 d) of the first 28 d under observation, and severe diarrhea for 7% (1.87 d) of the 28 d under observation. Using a repeated measures linear regression model, we found the proportion of days with diarrhea significantly decreased weight gain at 14, 56, and 77 d following arrival. An increased proportion of days with diarrhea increased the risk of mortality, which was determined using a Cox proportional hazards model. We also found, using 2 Cox proportional hazards models, that a higher proportion of days with an abnormal fecal score increased the hazard of antibiotic treatment. With respect to factors associated with the occurrence of abnormal fecal consistency, we found that arrival weight and the source of calves were statistically significant predictors. Specifically, for every additional kilogram of body weight at arrival, the proportion of days with diarrhea decreased by 7%. With respect to source, calves from drovers had a higher proportion of days with diarrhea compared with those sourced directly from local dairy farms. Our results highlight the substantial influence the presence of abnormal fecal consistency has on short-term weight gain, mortality risk, and morbidity risk. We also demonstrate that diarrhea occurrence can be predicted using body weight at arrival and calf source. Further research should evaluate longer-term effects of diarrhea and better understand the effect of arrival weight on neonatal calf disease risk.

Coordinated actions of SLX1-SLX4 and MUS81-EME1 for Holliday junction resolution in human cells.

Holliday junctions (HJs) are four-way DNA intermediates that form during homologous recombination, and their efficient resolution is essential for chromosome segregation. Here, we show that three structure-selective endonucleases, namely SLX1-SLX4, MUS81-EME1, and GEN1, define two pathways of HJ resolution in human cells. One pathway is mediated by GEN1, whereas SLX1-SLX4 and MUS81-EME1 provide a second and genetically distinct pathway (SLX-MUS). Cells depleted for SLX-MUS or GEN1 pathway proteins exhibit severe defects in chromosome segregation and reduced survival. In response to CDK-mediated phosphorylation, SLX1-SLX4 and MUS81-EME1 associate at the G2/M transition to form a stable SLX-MUS holoenzyme, which can be reconstituted in vitro. Biochemical studies show that SLX-MUS is a HJ resolvase that coordinates the active sites of two distinct endonucleases during HJ resolution. This cleavage reaction is more efficient and orchestrated than that mediated by SLX1-SLX4 alone, which exhibits a potent nickase activity that acts promiscuously upon DNA secondary structures.

Scurvy and Tinea Corporis Simulating Leukocytoclastic Vasculitis.

ABSTRACT: Leukocytoclastic vasculitis (LCV) is a small vessel inflammatory condition considered to be caused by circulating immune complexes and often occurs after an acute infection or exposure to a new medication, although it may be associated with an underlying systemic disease or be idiopathic in nature. It is important to determine the etiology, identify the extent of the disease for early intervention and appropriate management, and treat and/or eliminate the underlying cause. Here, we report cases of scurvy and tinea corporis that presented with histopathologic features of LCV and had significant clinical improvement with treatment of the underlying etiologies. These cases emphasize that histopathologic features of early evolving LCV may be seen in other settings including scurvy and tinea corporis. Appropriate treatment of the underlying condition is important for optimized patient management.

Beyond Muscle: A Narrative Review of Onabotulinum Toxin A for Skin Quality.

BACKGROUND: Onabotulinum toxin A’s (BoNTA’s) popularity is centered on its quick, predictable, and safe ability to improve clinically apparent rhytides. While this remains the only FDA indication for BoNTA, recent research suggests that BoNTA may have a much wider use for the improvement of overall skin quality. OBJECTIVE: This review will focus on the various evidence and uses of BoNTA for improvement of skin quality. MATERIALS AND METHODS: This review considered published journal articles (clinical trials, case studies, scientific reviews). Studies were identified by searching the PubMed database and reference lists of respective articles. Only articles available in English were considered for this review. RESULTS: Intramuscular and intradermal injections of BoNTA decrease sebum production, pore size, facial erythema and flushing, and resting lines. BoNTA induces neocollagenesis and restores the extracellular matrix. CONCLUSION: The literature provides both in vivo and in vitro evidence that BoNTA enhances skin quality beyond its well-studied neuromuscular effect. J Drugs Dermatol. 2021;20(9):958-962. doi:10.36849/JDD.6038.