Microcalcifications in benign breast biopsies: association with lesion type and risk.

PURPOSE: To characterize associations of microcalcifications (calcs) with benign breast disease lesion subtypes and assess whether tissue calcs affect risks of ductal carcinoma in situ (DCIS) and invasive breast cancer (IBC). METHODS: We analyzed detailed histopathologic data for 4,819 BBD biopsies from a single institution cohort (2002-2013) followed for DCIS or IBC for a median of 7.4 years for cases (N = 338) and 11.2 years for controls. Natural language processing was used to identify biopsies containing calcs based on pathology reports. Univariable and multivariable regression models were applied to assess associations with BBD lesion type and age-adjusted Cox proportional hazard regressions were performed to model risk of IBC or DCIS stratified by the presence or absence of calcs. RESULTS: Calcs were identified in 2063 (42.8%) biopsies. Calcs were associated with older age at BBD diagnosis (56.2 versus 49.0 years; P < 0.001). Overall, the risk of developing IBC or DCIS did not differ significantly between patients with calcs (HR 1.13, 95% CI 0.90, 1.41) as compared to patients without calcs. Stratification by BBD severity or subtype, age at BBD biopsy, outcomes of IBC versus DCIS, and mammography technique (screen-film versus full-field digital mammography) did not significantly alter association between calcs and risk. CONCLUSION: Our analysis of calcs in BBD biopsies did not find a significant association between calcs and risk of breast cancer.

Benign Breast Disease and Breast Cancer Risk in the Percutaneous Biopsy Era.

Importance: Benign breast disease (BBD) comprises approximately 75% of breast biopsy diagnoses. Surgical biopsy specimens diagnosed as nonproliferative (NP), proliferative disease without atypia (PDWA), or atypical hyperplasia (AH) are associated with increasing breast cancer (BC) risk; however, knowledge is limited on risk associated with percutaneously diagnosed BBD. Objectives: To estimate BC risk associated with BBD in the percutaneous biopsy era irrespective of surgical biopsy. Design, Setting, and Participants: In this retrospective cohort study, BBD biopsy specimens collected from January 1, 2002, to December 31, 2013, from patients with BBD at Mayo Clinic in Rochester, Minnesota, were reviewed by 2 pathologists masked to outcomes. Women were followed up from 6 months after biopsy until censoring, BC diagnosis, or December 31, 2021. Exposure: Benign breast disease classification and multiplicity by pathology panel review. Main Outcomes: The main outcome was diagnosis of BC overall and stratified as ductal carcinoma in situ (DCIS) or invasive BC. Risk for presence vs absence of BBD lesions was assessed by Cox proportional hazards regression. Risk in patients with BBD compared with female breast cancer incidence rates from the Iowa Surveillance, Epidemiology, and End Results (SEER) program were estimated. Results: Among 4819 female participants, median age was 51 years (IQR, 43-62 years). Median follow-up was 10.9 years (IQR, 7.7-14.2 years) for control individuals without BC vs 6.6 years (IQR, 3.7-10.1 years) for patients with BC. Risk was higher in the cohort with BBD than in SEER data: BC overall (standard incidence ratio [SIR], 1.95; 95% CI, 1.76-2.17), invasive BC (SIR, 1.56; 95% CI, 1.37-1.78), and DCIS (SIR, 3.10; 95% CI, 2.54-3.77). The SIRs increased with increasing BBD severity (1.42 [95% CI, 1.19-1.71] for NP, 2.19 [95% CI, 1.88-2.54] for PDWA, and 3.91 [95% CI, 2.97-5.14] for AH), comparable to surgical cohorts with BBD. Risk also increased with increasing lesion multiplicity (SIR: 2.40 [95% CI, 2.06-2.79] for ≥3 foci of NP, 3.72 [95% CI, 2.31-5.99] for ≥3 foci of PDWA, and 5.29 [95% CI, 3.37-8.29] for ≥3 foci of AH). Ten-year BC cumulative incidence was 4.3% for NP, 6.6% for PDWA, and 14.6% for AH vs an expected population cumulative incidence of 2.9%. Conclusions and Relevance: In this contemporary cohort study of women diagnosed with BBD in the percutaneous biopsy era, overall risk of BC was increased vs the general population (DCIS and invasive cancer combined), similar to that in historical BBD cohorts. Development and validation of pathologic classifications including both BBD severity and multiplicity may enable improved BC risk stratification.

Relevance of comorbidities and antithrombotic medication as risk factors for reoperation in patients with chronic subdural hematoma.

In an aging Western society, the incidence of chronic subdural hematomas (cSDH) is continuously increasing. In this study, we reviewed our clinical management of cSDH patients and identified predictive factors for the need of reoperation due to residual or recurrent hematomas with a focus on the use of antithrombotic drugs. In total, 623 patients who were treated for cSDH with surgical evacuation between 2006 and 2016 at our department were retrospectively analyzed. Clinical and radiological characteristics and laboratory parameters were investigated as possible predictors of reoperation with univariate and multivariate analyses. Additionally, clinical outcome measures were compared between patients on anticoagulants, on antiplatelets, and without antithrombotic medication. In univariate analyses, patients on anticoagulants and antiplatelets presented significantly more often with comorbidities, were significantly older, and their risk for perioperative complications was significantly increased. Nevertheless, their clinical outcome was comparable to that of patients without antithrombotics. In multivariate analysis, only the presence of comorbidities, but not antithrombotics, was an independent predictor for the need for reoperations. Patients on antithrombotics do not seem to necessarily have a significantly increased risk for residual hematomas or rebleeding requiring reoperation after cSDH evacuation. More precisely, the presence of predisposing comorbidities might be a key independent risk factor for reoperation. Importantly, the clinical outcomes after surgical evacuation of cSDH are comparable between patients on anticoagulants, antiplatelets, and without antithrombotics.

A Recombinant Protein SARS-CoV-2 Candidate Vaccine Elicits High-titer Neutralizing Antibodies in Macaques.

Background: Vaccines that generate robust and long-lived protective immunity against SARS-CoV-2 infection are urgently required. Methods: We assessed the potential of vaccine candidates based on the SARS-CoV-2 spike in cynomolgus macaques (M. fascicularis) by examining their ability to generate spike binding antibodies with neutralizing activity. Antigens were derived from two distinct regions of the spike S1 subunit, either the N-terminal domain or an extended C-terminal domain containing the receptor-binding domain and were fused to the human IgG1 Fc domain. Three groups of 2 animals each were immunized with either antigen, alone or in combination. The development of antibody responses was evaluated through 20 weeks post-immunization. Results: A robust IgG response to the spike protein was detected as early as 2 weeks after immunization with either protein and maintained for over 20 weeks. Sera from animals immunized with antigens derived from the RBD were able to prevent binding of soluble spike proteins to the ACE2 receptor, shown by in vitro binding assays, while sera from animals immunized with the N-terminal domain alone lacked this activity. Crucially, sera from animals immunized with the extended receptor binding domain but not the N-terminal domain had potent neutralizing activity against SARS-CoV-2 pseudotyped virus, with titers in excess of 10,000, greatly exceeding that typically found in convalescent humans. Neutralizing activity persisted for more than 20 weeks. Conclusions: These data support the utility of spike subunit-based antigens as a vaccine for use in humans.

Effect of drug disposal bag provision on families' disposal of children's unused opioids.

OBJECTIVE: One strategy to combat opioid misuse is to remove excess opioids from circulation by providing patients with drug disposal products that enable the safe disposal of opioids. We aimed to evaluate opioid use and disposal of unused opioids among children and young adults before and after pharmacy staff at our institution began to provide patients and families filling opioid prescriptions with a drug disposal bag. METHODS: We performed a prospective pre-post cohort study of patients who filled an opioid prescription in May-August 2019 at the outpatient pharmacies of a large tertiary children's hospital. Patients and caregivers were enrolled at the time the opioids were dispensed. During the first half of the study period, standard opioid-related education was offered by pharmacy staff. During the second half of the study period, standard education was offered, and a drug disposal bag and instructions on its use were provided when the opioids were dispensed. A follow-up survey to assess opioid use and disposal was completed online or by telephone 4-7 weeks after the opioids were dispensed. RESULTS: A total of 215 participants were enrolled; 117 received a drug disposal bag and 98 did not. Of those, 68% of the participants completed a follow-up survey. In both groups, the median patient age was 11 years, and most patients had been prescribed opioids after a procedure. More than 70% had opioids leftover after they had stopped taking them, and this did not vary by group. However, among families with leftover opioids, the receipt of a drug disposal bag was associated with a higher likelihood of disposal of the unused opioids (71.7% vs. 52.1%, P = 0.04). CONCLUSION: Providing a drug disposal bag to families of children receiving opioids increases the likelihood of excess opioid disposal. Greater availability of drug disposal products can complement prescribing reduction efforts aimed at decreasing prescription opioid misuse.

Mortality and functional outcome after surgical evacuation of traumatic acute subdural hematomas in octa- and nonagenarians.

PURPOSE: The incidence of acute subdural hematomas (aSDH) is rising. However, beneficial effects of surgery for the oldest aSDH patients remain unclear. We hence describe the postoperative outcome of octa- and nonagenarians with aSDH in comparison to a younger patient cohort. METHODS: Patients aged ≥ 80 years surgically treated for traumatic aSDH at a single institution between 2006 and 2016 were retrospectively reviewed. Clinical and imaging variables were assessed, and univariate analysis was performed to identify factors predicting outcome at discharge. Results were compared to a cohort of younger aSDH patients and statistical analysis was performed. Long-term outcome was prospectively evaluated with the GOSE and QOLIBRI. RESULTS: 27 aSDH patients aged ≥ 80 years were identified. On admission, 41% were in a comatose state and in-hospital mortality was 33%. At discharge, 22% had a favorable outcome (GOS 4 + 5). In univariate statistical analysis, better neurological status (GCS > 8), ≤ 1 comorbidity and smaller aSDH volumes were significant predictors for a favorable outcome. Comparison to 27 younger aSDH patients revealed significant differences in the prevalence of comorbidities and antithrombotics. At long-term follow-up, quality of life of aSDH patients was reduced (median QOLIBRI 54%). CONCLUSION: Outcome after surgical treatment of aSDH in octa- and nonagenarians is not detrimental per se. Predictors for a favorable outcome are a non-comatose state on admission (GCS > 8), ≤ 1 preexisting comorbidity and a lower aSDH volume in patients aged ≥ 80 years. In individual patients, surgical evacuation of aSDH might remain a treatment option even in high ages.

Economic impact of pharmacist interventions in pediatric ambulatory care clinics.

OBJECTIVES: Ambulatory care pharmacists have a unique opportunity to identify and prevent adverse drug events (ADEs) throughout a patient's treatment course. These interventions can reduce unexpected clinic visits or hospitalizations, which may lead to decreased health care costs. However, research on this topic has not been conducted in the pediatric population. This study explored the economic impact of pharmacist interventions related to ADEs in pediatric ambulatory care clinics. The primary objective was to determine the total cost avoidance of pharmacist interventions associated with the prevention or management of ADEs in pediatric ambulatory care clinics. The secondary objectives were to describe and quantify pharmacist interventions related to the prevention and management of ADEs in pediatric ambulatory care clinics. METHODS: Pharmacist interventions from pediatric ambulatory care clinics were collected from an electronic health record. These interventions were categorized into 1 of 4 categories: Drug interaction, drug not indicated, prevent or manage ADE, or prevent or manage drug allergy. A review panel consisting of ambulatory care pharmacists reviewed the interventions. The expected probability of the event occurring was classified according to the Nesbit method (0-0.6), and the level of care necessary to treat the potential ADE was determined. The levels of care included hospitalization, ambulatory care, and self-care. The cost avoidance associated with each prevented ADE was calculated by multiplying the probability of the ADE occurring by the average charge of the expected level of care. RESULTS: Of the 8755 interventions documented, 212 were included, leading to a total cost avoidance of $307,210 (range $76,802-$1,071,053). The estimated cost avoidance from each ADE subtype was $128,283 from drug interaction, $20,727 from drug not indicated, $157,993 from prevent or manage ADE, and $207 from prevent or manage drug allergy. CONCLUSION: Pediatric ambulatory care pharmacists optimize health care cost savings through the prevention and management of ADEs as integrated members of the health care team.

Evaluation of medication safety resources in pediatric hospitals.

PURPOSE: As health systems continue to expand pharmacy and clinical services, the ability to evaluate potential medication safety risks and mitigate errors remains a high priority. Workload and productivity monitoring tools for the assessment of operational and clinical pharmacy services exist. However, such tools are not currently available to justify medication safety pharmacy services. The purpose of this study is to determine methods used to assess, allocate, and justify medication safety resources in pediatric hospitals. METHODS: A 32-question survey was designed and distributed utilizing the Research Electronic Data Capture (REDCap) tool. The survey was disseminated to 46 pediatric hospitals affiliated with the Children's Hospital Association (CHA). The survey was distributed in October 2018, and the respondents were given 3 weeks to submit responses. Data analysis includes the use of descriptive statistics. Categorical variables were summarized by frequencies and percentages to distinguish the differences between pediatric health systems. RESULTS: Of 26 respondents, 15.4% utilized metrics to justify medication safety resources. Metrics utilized were based on medication dispenses, projects, and error coding. Twenty-three percent of respondents were dissatisfied with current pharmacy-based medication safety resources within the organization. There was variability of medication safety resources within pediatric hospitals, including the number of dedicated full-time equivalents, time spent on tasks, and task prioritization. CONCLUSION: Assessing medication safety resources at various pediatric hospitals highlights several potential barriers and opportunities. This information will serve as the foundation for the creation of a standardized workload assessment tool to assist pharmacy leaders with additional resource justification.

"Inflammaging" and bone in the OsteoLaus cohort.

BACKGROUND: "Inflammaging" is a coined term that combines the processes of inflammation (within the normal range) and aging, since chronic, low-grade, systemic inflammation emerges with increasing age. Unlike high-level inflammation, with which deleterious effects on bone no longer need to be demonstrated, it is unclear whether inflammaging exerts deleterious effects on bone too. METHOD: We assessed associations between inflammaging - measured via cytokine levels (high-sensitivity C-reactive protein (hs-CRP); interleukin-1ß (IL-1ß); interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α)) - and bone parameters (prevalent and incident fractures, bone mineral density (BMD) and trabecular bone score (TBS)) in 1390 postmenopausal women from the OsteoLaus study. RESULTS: Mean (±SD) age was 64.5 ± 7.6 and mean bone mass index (BMI) 25.9 ± 4.5 kg/m2. Median hs-CRP, IL-1ß, IL-6 and TNF-α were 1.4 pg/ml, 0.57 pg/ml, 2.36 pg/ml and 4.82 pg/ml, respectively. In total, 10.50% of the participants had a prevalent, low-impact fracture; and, after 5-years of follow up, 5.91% had an incident, low-impact fracture. Mean T-score BMD was - 1.09 ± 1.53 for the spine, - 1.08 ± 1.02 for the femoral neck, and - 0.72 ± 0.96 for the total hip. Mean spine TBS was 1.320 ± 0.10. We found a positive association between hs-CRP and BMD at all sites, and between hs-CRP and the TBS, but none of these associations were significant after adjustment. We found no association between prevalent or incident fractures and hs-CRP. No association was found between IL-1ß, IL6 and TNF-α and BMD, TBS or fractures. CONCLUSION: Our results suggest that bone imaging and structure parameters are not associated with the low-grade cytokine levels (within the normal range) observed with inflammaging.

"A recombinant protein SARS-CoV-2 candidate vaccine elicits high-titer neutralizing antibodies in macaques."

Vaccines that generate robust and long-lived protective immunity against SARS-CoV-2 infection are urgently required. We assessed the potential of vaccine candidates based on the SARS-CoV-2 spike in cynomolgus macaques (M. fascicularis) by examining their ability to generate spike binding antibodies with neutralizing activity. Antigens were derived from two distinct regions of the spike S1 subunit, either the N-terminal domain (NTD) or an extended C-terminal domain containing the receptor-binding domain (RBD) and were fused to the human IgG1 Fc domain. Three groups of 2 animals each were immunized with either antigen, alone or in combination. The development of antibody responses was evaluated through 20 weeks post-immunization. A robust IgG response to the spike protein was detected as early as 2 weeks after immunization with either protein and maintained for over 20 weeks. Sera from animals immunized with antigens derived from the RBD were able to prevent binding of soluble spike proteins to the ACE2 receptor, shown by in vitro binding assays, while sera from animals immunized with the NTD alone lacked this activity. Crucially, sera from animals immunized with the RBD but not the NTD had potent neutralizing activity against SARS-CoV-2 pseudotyped virus, with titers in excess of 10,000, greatly exceeding that typically found in convalescent humans. Neutralizing activity persisted for more than 20 weeks. These data support the utility of spike subunit-based antigens as a vaccine for use in humans.

The influence of gravity and light on locomotion and orientation of Heterocypris incongruens and Notodromas monacha (Crustacea, Ostracoda).

For future manned long-d uration space missions, the supply of essentials, such as food, water, and oxygen with the least possible material resupply from Earth is vital. This need could be satisfied utilizing aquatic bioregenerative life support systems (BLSS), as they facilitate recycling and autochthonous production. However, few organisms can cope with the instable environmental conditions and organic pollution potentially prevailing in such BLSS. Ostracoda, however, occur in eu- and even hypertrophic waters, tolerate organic and chemical waste, varying temperatures, salinity, and pH ranges. Thus, according to their natural role, they can link oxygen liberating, autotrophic algae, and higher trophic levels (e.g., fish) probably also in such harsh BLSS. Yet, little is known about how microgravity (µg) affects Ostracoda. In this regard, we investigated locomotion and orientation, as they are involved in locating mating partners and suitable microhabitats, foraging, and escaping predators. Our study shows that Ostracoda exhibit altered activity patterns and locomotion behavior (looping) in µg. The alterations are differentially marked between the studied species (i.e., 2% looping in Notodromas monacha, ~50% in Heterocypris incongruens) and also the thresholds of gravity perception are distinct, although the reasons for these differences remain speculative. Furthermore, neither species acclimates to µg nor orientates by light in µg. However, Ostracoda are still promising candidates for BLSS due to the low looping rate of N. monacha and our findings that the so far analyzed vital functions and life-history parameters of H. incongruens remained similar as under normal gravity conditions despite of its high looping rate.

p38MAPK/MK2-dependent phosphorylation controls cytotoxic RIPK1 signalling in inflammation and infection.

Receptor-interacting protein kinase-1 (RIPK1), a master regulator of cell fate decisions, was identified as a direct substrate of MAPKAP kinase-2 (MK2) by phosphoproteomic screens using LPS-treated macrophages and stress-stimulated embryonic fibroblasts. p38MAPK/MK2 interact with RIPK1 in a cytoplasmic complex and MK2 phosphorylates mouse RIPK1 at Ser321/336 in response to pro-inflammatory stimuli, such as TNF and LPS, and infection with the pathogen Yersinia enterocolitica. MK2 phosphorylation inhibits RIPK1 autophosphorylation, curtails RIPK1 integration into cytoplasmic cytotoxic complexes, and suppresses RIPK1-dependent apoptosis and necroptosis. In Yersinia-infected macrophages, RIPK1 phosphorylation by MK2 protects against infection-induced apoptosis, a process targeted by Yersinia outer protein P (YopP). YopP suppresses p38MAPK/MK2 activation to increase Yersinia-driven apoptosis. Hence, MK2 phosphorylation of RIPK1 is a crucial checkpoint for cell fate in inflammation and infection that determines the outcome of bacteria-host cell interaction.

Olfactory brain gray matter volume reduction in patients with chronic rhinosinusitis.

BACKGROUND: Chronic rhinosinusitis (CRS) is a common inflammatory condition and a major cause of olfactory loss. Olfactory dysfunction has been associated with reduced olfactory bulb (OB) volume and gray matter (GM) density in the olfactory-related brain areas. The aim of this study was to investigate brain GM structural and OB volume alterations in patients with CRS. METHODS: Structural brain images were collected from 21 CRS patients and 31 healthy controls on a 3-T scanner. Voxel-based morphometry (VBM) was performed to investigate GM. Olfactory bulb volumes were measured using AMIRA software. Psychophysical olfactory testing for odor threshold (T) and identification (I) was performed using the Sniffin' Sticks battery. RESULTS: CRS patients had significantly lower scores for Sniffin' Sticks olfactory tests than controls (p < 0.001 for T, I, and combined T and I [TI] scores). Region-of-interest analyses revealed no difference in GM volume between CRS patients and healthy controls; however, in CRS patients with severe olfactory dysfunction, GM reduction was observed in the gyrus rectus, orbitofrontal cortex, thalamus, and the insula. In addition, no difference was observed for OB volume in CRS patients compared with healthy controls. CONCLUSION: In this study we identified a reduction in gray matter in olfactory brain regions in CRS patients with severe olfactory dysfunction.

Multidisciplinary Optimization of Oral Chemotherapy Delivery at the University of Wisconsin Carbone Cancer Center.

PURPOSE: Use of oral chemotherapy is expanding and offers advantages while posing unique safety challenges. ASCO and the Oncology Nursing Society jointly published safety standards for administering chemotherapy that offer a framework for improving oral chemotherapy practice at the University of Wisconsin Carbone Cancer Center. METHODS: With the goal of improving safety, quality, and uniformity within our oral chemotherapy practice, we conducted a gap analysis comparing our practice against ASCO/Oncology Nursing Society guidelines. Areas for improvement were addressed by multidisciplinary workgroups that focused on education, workflows, and information technology. Recommendations and process changes included defining chemotherapy, standardizing patient and caregiver education, mandating the use of comprehensive electronic order sets, and standardizing documentation for dose modification. Revised processes allow pharmacists to review all orders for oral chemotherapy, and they support monitoring adherence and toxicity by using a library of scripted materials. RESULTS: Between August 2015 and January 2016, revised processes were implemented across the University of Wisconsin Carbone Cancer Center clinics. The following are key performance indicators: 92.5% of oral chemotherapy orders (n = 1,216) were initiated within comprehensive electronic order sets (N = 1,315), 89.2% compliance with informed consent was achieved, 14.7% of orders (n = 193) required an average of 4.4 minutes review time by the pharmacist, and 100% compliance with first-cycle monitoring of adherence and toxicity was achieved. CONCLUSION: We closed significant gaps between institutional practice and published standards for our oral chemotherapy practice and experienced steady improvement and sustainable performance in key metrics. We created an electronic definition of oral chemotherapies that allowed us to leverage our electronic health records. We believe our tools are broadly applicable.

Single-Dose Basiliximab Induction in Low-Risk Renal Transplant Recipients.

OBJECTIVES: To compare the efficacy of a single dose of basiliximab with two doses in preventing acute rejection in selected low-risk renal transplant recipients. METHODS: This observational study of 760 kidney transplant recipients considered to be at low immunologic risk (peak panel reactive antibody less than 10%) compared patient and graft outcomes following a single-dose versus a two-dose regimen of basiliximab. MAIN RESULTS: No differences were found in patient survival (92% vs 92%, p=0.6), graft survival (86% vs 83%, p=0.2), acute rejection (cellular [4% vs 7%, p=0.2], antibody-mediated rejection [19% vs 19%, p=0.9]), or opportunistic infections (34% vs 30%, p=0.3) between the single versus two-dose regimens, respectively. In multivariate analyses, the number of doses of basiliximab was not associated with acute rejection or patient/graft survival despite adjustment with Cox regression and propensity scores. However, delayed graft function (DGF), donor age older than 65 years, and human leukocyte antigen mismatch of 3 or higher were associated with acute rejection (hazard ratio [HR] 2.64, 1.91, and 1.57, respectively, p≤0.04), and DGF and diabetes were associated with death/graft loss (HR 2.56 and 1.63, respectively, p≤0.009). PRINCIPAL CONCLUSIONS: A single dose of basiliximab is safe and effective for induction in low-risk kidney transplant recipients.

Linker Nups connect the nuclear pore complex inner ring with the outer ring and transport channel.

Nuclear pore complexes (NPCs) mediate transport between the nucleus and cytoplasm. NPCs are composed of ∼30 nucleoporins (Nups), most of which are organized in stable subcomplexes. How these modules are interconnected within the large NPC framework has been unknown. Here we show a mechanism of how supercomplexes can form between NPC modules. The Nup192 inner-pore-ring complex serves as a seed to which the Nup82 outer-ring complex and Nsp1 channel complex are tethered. The linkage between these subcomplexes is generated by short sequences within linker Nups. The conserved Nup145N is the most versatile connector of NPC modules because it has three discrete binding sites for Nup192, Nup170 and Nup82. We assembled a large part of a Chaetomium thermophilum NPC protomer in vitro, providing a step forward toward the reconstitution of the entire NPC.

Architecture of the fungal nuclear pore inner ring complex.

The nuclear pore complex (NPC) constitutes the sole gateway for bidirectional nucleocytoplasmic transport. We present the reconstitution and interdisciplinary analyses of the ~425-kilodalton inner ring complex (IRC), which forms the central transport channel and diffusion barrier of the NPC, revealing its interaction network and equimolar stoichiometry. The Nsp1•Nup49•Nup57 channel nucleoporin heterotrimer (CNT) attaches to the IRC solely through the adaptor nucleoporin Nic96. The CNT•Nic96 structure reveals that Nic96 functions as an assembly sensor that recognizes the three-dimensional architecture of the CNT, thereby mediating the incorporation of a defined CNT state into the NPC. We propose that the IRC adopts a relatively rigid scaffold that recruits the CNT to primarily form the diffusion barrier of the NPC, rather than enabling channel dilation.

Structural basis for assembly and function of the Nup82 complex in the nuclear pore scaffold.

Nuclear pore complexes (NPCs) are huge assemblies formed from ∼30 different nucleoporins, typically organized in subcomplexes. One module, the conserved Nup82 complex at the cytoplasmic face of NPCs, is crucial to terminate mRNA export. To gain insight into the structure, assembly, and function of the cytoplasmic pore filaments, we reconstituted in yeast the Nup82-Nup159-Nsp1-Dyn2 complex, which was suitable for biochemical, biophysical, and electron microscopy analyses. Our integrative approach revealed that the yeast Nup82 complex forms an unusual asymmetric structure with a dimeric array of subunits. Based on all these data, we developed a three-dimensional structural model of the Nup82 complex that depicts how this module might be anchored to the NPC scaffold and concomitantly can interact with the soluble nucleocytoplasmic transport machinery.

Comparison of isoflurane inhalation anaesthesia, injection anaesthesia and high volume caudal epidural anaesthesia for umbilical surgery in calves; metabolic, endocrine and cardiopulmonary effects.

OBJECTIVE: To compare three anaesthetic protocols for umbilical surgery in calves regarding adequacy of analgesia, and cardiopulmonary and hormonal responses. STUDY DESIGN: Prospective, randomised experimental study. ANIMALS: Thirty healthy German Holstein calves (7 female, 23 male) aged 45.9 ± 6.4 days. METHODS: All calves underwent umbilical surgery in dorsal recumbency. The anaesthetic protocols were as follows: group INH (n = 10), induction 0.1 mg kg(-1) xylazine IM and 2.0 mg kg(-1) ketamine IV, maintenance isoflurane in oxygen; Group INJ (n = 10), induction 0.2 mg kg(-1) xylazine IM and 5.0 mg kg(-1) ketamine IV, maintenance 2.5 mg kg(-1) ketamine IV every 15 minutes or as required; group EPI (n = 10), high volume caudal epidural anaesthesia with 0.2 mg kg(-1) xylazine diluted to 0.6 mL kg(-1) with procaine 2%. All calves received peri-umbilical infiltration of procaine and pre-operative IV flunixin (2.2 mg kg(-1) ). Cardiopulmonary variables were measured at preset intervals for up to 2 hours after surgery. The endocrine stress response was determined. Intra-operative nociception was assessed using a VAS scale. Data were compared between groups using appropriate statistical tests. A value of p < 0.05 was considered significant. RESULTS: All three protocols provided adequate anaesthesia for surgery although, as judged by the VAS scale, intra-operative response was greatest with INJ. Lowest mean cortisol levels during surgery occurred in EPI. Heart rate and cardiac output did not differ between groups, but mean arterial blood pressure, systemic vascular resistance, and partial pressure of carbon dioxide were higher and arterial pH lower in groups INH and INJ than in Group EPI. Group INJ became hypoxaemic and had a significantly greater vascular shunt than did the other groups. CONCLUSION AND CLINICAL RELEVANCE: Groups INH and EPI both proved acceptable protocols for calves undergoing umbilical surgery, whilst INJ resulted in variable anti-nociception and in hypoxaemia. High volume caudal epidural anaesthesia provides a practical inexpensive method of anaesthesia for umbilical surgery.