The John S. Najarian symposium: The past, present, and future of surgery and transplantation, May 20, 2022, Minneapolis, MN.

Dr John S Najarian (1927-2020), chairman of the Department of Surgery at the University of Minnesota from 1967 to 1993, was a pioneer in surgery, clinical immunology and transplantation. A Covid-delayed Festschrift was held in his honor on May 20, 2022. The speakers reflected on his myriad contributions to surgery, transplantation, and resident/fellow training, as well as current areas of ongoing research to improve clinical outcomes. Of note, Dr Najarian was a founder of the journal Clinical Transplantation.

Structural Elucidation of Peptide Binding to KLHL-12, a Substrate Specific Adapter Protein in a Cul3-Ring E3 Ligase Complex.

KLHL-12 is a substrate specific adapter protein for a Cul3-Ring ligase complex. It is a member of the Kelch ß-propeller domain subclass of Cullin-Ring substrate recognition domains. This E3 ubiquitin ligase complex has many activities, including acting as a negative regulator of the Wnt signaling pathway by mediating ubiquitination and subsequent proteolysis of Dvl3/Dsh3. KLHL-12 is also known to mediate the polyubiquitination of the dopamine D4 receptor (D4.2), the ubiquitination of KHSRP, a protein that is involved in IRES translation, and also the ubiquitination of Sec31, which is involved in endoplasmic reticulum-Golgi transport by regulating the size of COPII coats. Earlier studies broadly defined the substrate binding regions for D4.2 and Dvl3/Dsh3 to KLHL-12. We tested several peptides from these regions and succeeded in identifying a short peptide that bound to KLHL-12 with low micromolar affinity. To better understand the sequence specificity of this peptide, we used alanine substitutions to map the important residues and obtained an X-ray structure of this peptide bound to KLHL-12. This structure and our peptide affinity measurements suggest a sequence motif for peptides that bind to the top face of KLHL-12. Understanding this binding site on KLHL-12 may contribute to efforts to find small molecule ligands that can either directly inhibit the degradation of substrate proteins or be used in targeted protein degradation strategies using PROTACs.

ω-Hydroxy isoprenoid bisphosphonates as linkable GGDPS inhibitors.

The enzyme geranylgeranyl diphosphate synthase (GGDPS) is a potential therapeutic target for multiple myeloma. Malignant plasma cells produce and secrete large amounts of monoclonal protein, and inhibition of GGDPS results in disruption of protein geranylgeranylation which in turn impairs intracellular protein trafficking. Our previous work has demonstrated that some isoprenoid triazole bisphosphonates are potent and selective inhibitors of GGDPS. To explore the possibility of selective delivery of such compounds to plasma cells, new analogues with an ω-hydroxy group have been synthesized and examined for their enzymatic and cellular activity. These studies demonstrate that incorporation of the ω-hydroxy group minimally impairs GGDPS inhibitory activity. Furthermore conjugation of one of the novel ω-hydroxy GGDPS inhibitors to hyaluronic acid resulted in enhanced cellular activity. These results will allow future studies to focus on the in vivo biodistribution of HA-conjugated GGDPS inhibitors.

Probing the self-assembly dynamics and internal structure of amphiphilic hyaluronic acid conjugates by fluorescence spectroscopy and molecular dynamics simulations.

Polymeric nanoparticles are increasingly used as biocompatible carriers for drugs and imaging agents. Understanding their self-assembly dynamics and morphology is of ultimate importance to develop nanoformulations with optimal characteristics. To achieve better performance, it is vital to account for cargo-carrier interactions at the molecular level. The self-assembly dynamics were studied and the internal structure of nanoparticles derived from a series of hydrophobically modified hyaluronic acid was revealed. Environment-sensitive ratiometric fluorescent probes provide valuable information about the nanoparticle's interior morphology, and molecular dynamics simulations complement the overall picture with insights into intramolecular and intermolecular interactions of the polymer, as well as its interactions with the small-molecule load. van der Waals and π-π interactions of the hydrophobic side fragments play a leading role in self-assembly and loading of hydrophobic small molecules. Aliphatic substituents form more extensive hydrophobic domains, while aromatic moieties allow more interaction of the loaded small molecules with the surrounding solvent.

Development of colloidally stable carbazole-based fluorescent nanoaggregates.

Fluorescent nanomaterials require high colloidal stability for effective use in imaging and sensing applications. We herein report the synthesis of carbazole-based organic fluorescent nanoaggregates, and demonstrate the superior colloidal stability of alkyl-substituted dye aggregates over their non-alkylated analogs. The role of alkyl chains in self-assembly and stability of such nanoaggregates is discussed based on both experimental and molecular dynamics simulation data, and spectral characteristics of the precursor dyes and their aggregates are described. The obtained results provide new insights on development of colloidally stable organic fluorescent nanomaterials with low polydispersity.

In vitro fungicidal effects of methylene blue at 625-nm.

The aim of the study is to confirm the effectiveness of photodynamic therapy (PDT) as a significant inhibitor of Trichophyton rubrum (T. rubrum) and to determine the most appropriate dose and rate of delivery. Trichophyton rubrum is the most common dermatophyte worldwide, responsible for the majority of superficial fungal infections. The traditional treatment of T. rubrum has known adverse effects. An alternative treatment is warranted. Photosensitised T. rubrum specimens were treated with 625-nm light at doses of 3, 12, 24, 40 and 60 J/cm2 . Colony counts were performed and compared to untreated controls. Doses of 24, 40 and 60 J/cm2 all produced kill rates of over 94%. A lower rate of delivery (7.80 mW/cm2 ) was shown to be a greater inhibitor of T. rubrum than a higher rate of delivery (120 mW/cm2 ). Photodynamic therapy with methylene blue (MB) at 625 nm using a low rate of delivery at doses of 24, 40 and 60 J/cm2 is an effective inhibitor of T. rubrum. A rate of delivery of 7.80 mW/cm2 is a significantly greater inhibitor of T. rubrum than a rate of 120 mW/cm2 when applying 625-nm light in PDT using MB.

Susceptibility of Trichopyton mentagrophytes to Visible Light Wavelengths.

OBJECTIVE: To determine whether a blue light (405 nm) could inhibit the growth of Trichopyton mentagrophytes without using a photosensitizing material as part of the treatment protocol. DESIGN: Basic physiologic randomized trial using laboratory specimens (T mentagrophytes). INTERVENTIONS/METHODS: Plated on a growth medium, T mentagrophytes were exposed to 3 to 5 administrations of blue light at 20 J/cm over 28 hours. Following 7 days of incubation, colony-forming units were counted and compared with nonirradiated controls. RESULTS: The study found 3, 4, and 5 administrations of blue light produced significant inhibition of T mentagrophytes (P < .05); 4 and 5 applications produced the greatest inhibition of growth (84.7% and 93.6% kill rates, respectively). CONCLUSIONS: The application of 405-nm light at a dose of 20 J/cm is an effective in vitro inhibitor of T mentagrophytes. To give results similar to those seen when a photosensitizing material is included, 3 to 5 applications of this wavelength and dose condition delivered over 28 hours is likely needed.

Longterm health-related quality of life after living liver donation.

There are little data on longterm outcomes, health-related quality of life (HRQoL), and issues related to living donor right hepatectomy specifically. We studied longterm HRQoL in 127 living liver donors. A donor-specific survey (DSS) was used to evaluate the living liver donor morbidity, and the 36-item short-form health survey (short-form 36 health survey, version 1 [SF-36]) was used to assess generic outcomes. The DSS was completed by 107 (84.3%) donors and the SF-36 by 62 (49%) donors. Median follow-up was 6.9 years. Of the 107 donors, 12 (11.2%) donors reported their health as better, whereas 84 (78.5%) reported their health the same as before donation. Ninety-seven (90.7%) are currently employed. The most common postdonation symptom was incisional discomfort (34%). Twenty-four donors (22.4%) self-reported depression symptoms after donation. Ninety-eight (91.6%) rated their satisfaction with the donation process ≥ 8 (scale of 1-10). Three factors-increased vitality (correlation, 0.44), decreased pain (correlation, 0.34), and a recipient who was living (correlation, 0.44)-were independently related to satisfaction with the donor experience. Vitality showed the strongest association with satisfaction with the donor experience. Mental and physical component summary scale scores for donors were statistically higher compared to the US population norm (P < 0.001). Donors reported a high satisfaction rate with the donation process, and almost all donors (n = 104, 97.2%) would donate again independent of experiencing complications. Our study suggests that over a longterm period, liver donors continue to have above average HRQoL compared to the general population.

Inactivation of Candida albicans following exposure to 624-nanometer light from a supraluminous diode array.

OBJECTIVE: To determine the potential for visible light (405 or 624 nm) to produce an inhibitory effect on Candida albicans. In addition, the study sought to evaluate a series of doses in terms of their respective inhibiting capabilities. BACKGROUND DATA: The authors have studied the effect of blue light on Staphylococcus aureus and found that a bactericidal outcome can be obtained with low doses of blue light. METHODS: Candida albicans was tested because of its common appearance in human skin and mucous membrane infections. The organism was treated in vitro with 405-nm (blue) and with 624-nm (red) light emitted from a supraluminous diode array. Doses of 3, 9, 15, 30, and 60 J/cm(2) were used. Colony counts were performed and compared with untreated controls using Student t tests and 1-way analysis of variance with Tukey post hoc analysis. RESULTS: The results revealed no inhibition produced by 405 nm on C albicans (F4,20 = 0.901; P = .482). However, 624 nm did inhibit growth of C albicans at 3, 9, and 30 J/cm(2) (F4,20 = 6.064; P = .002). CONCLUSIONS: Appropriate doses of 624-nm light from a supraluminous diode array can inhibit the growth of C albicans in vitro. Three, 9, and 30 J/cm(2) are all effective dose levels.

Multi-platform next-generation sequencing of the domestic turkey (Meleagris gallopavo): genome assembly and analysis.

A synergistic combination of two next-generation sequencing platforms with a detailed comparative BAC physical contig map provided a cost-effective assembly of the genome sequence of the domestic turkey (Meleagris gallopavo). Heterozygosity of the sequenced source genome allowed discovery of more than 600,000 high quality single nucleotide variants. Despite this heterozygosity, the current genome assembly (∼1.1 Gb) includes 917 Mb of sequence assigned to specific turkey chromosomes. Annotation identified nearly 16,000 genes, with 15,093 recognized as protein coding and 611 as non-coding RNA genes. Comparative analysis of the turkey, chicken, and zebra finch genomes, and comparing avian to mammalian species, supports the characteristic stability of avian genomes and identifies genes unique to the avian lineage. Clear differences are seen in number and variety of genes of the avian immune system where expansions and novel genes are less frequent than examples of gene loss. The turkey genome sequence provides resources to further understand the evolution of vertebrate genomes and genetic variation underlying economically important quantitative traits in poultry. This integrated approach may be a model for providing both gene and chromosome level assemblies of other species with agricultural, ecological, and evolutionary interest.

Removal of an infected aortic endograft and open aortic reconstruction: technical remarks.

Infection of an aortic endoprosthesis is a potentially lethal complication of an endovascular abdominal aortic aneurysm repair (EVAR). Surgical treatment usually involves excision of the infected endograft and vascular reconstruction, either in-line or extraanatomic. We present a case of severe infection of an endograft with suprarenal fixation. The one-stage procedure describes an in-line aortic reconstruction using a hybrid allograft composed of cryopreserved segments of cadaveric thoracic aorta and superficial femoral vein. A novel device for the removal of suprarenal fixating struts is described, as well as other technical maneuvers related to the removal of the struts used by surgeons in the community. The technique and surgical approach should be individualized based on patient characteristics, preoperative imaging, and knowledge of endograft construction.

Percutaneous atrial septal defect closure in infants and toddlers: predictors of success.

Device closure of atrial septal defect (ASD) is commonly performed in older children and adults. Infants and toddlers (age <4 years) are seldom referred for ASD closure due to size constraints. However, in many cases device ASD closure can be performed in this population. Between 2002 and 2012, 61 infants and toddlers were taken to the catheterization laboratory at our institution for ASD closure. Precatheterization transthoracic echocardiograms, intracatheterization transesophageal echocardiograms, and catheterization reports were reviewed. Fifty-three infants and toddlers presented for percutaneous ASD occlusion. Forty-eight (79 %) underwent successful closure, and 13 were referred for surgery without device attempt (n = 8) or after unsuccessful device occlusion (n = 4). Median age and weight at time of ASD closure were 2.99 years (range 0.3-3.8) and 11.7 kg (range 3.7-16.5). The device-to-septal length ratio was 0.81 (range 0.44-1.03). The 12 unsuccessful cases occurred in patients with larger defects (ASD diameter 17.5 ± 6.1 vs. 12.1 ± 4.2, p < 0.01). Deficient rims (absent or ≤ 4 mm) were seen in 9 of 12 (75 %) unsuccessful cases and in 19 of 41 (46 %) successful cases (p = 0.12). Multivariate analysis showed that patient size and ASD size were not independently associated with procedural success but that ASD size-to-patient weight ratio <1.2 (hazard ratio 9.5 [range1.7-17]) was associated with successful ASD closure. ASD device occlusion can be safely achieved in small children. An ASD size-to-patient weight ratio >1.2, not absolute patient weight or age, is associated with failure of the percutaneous approach. The midterm outcomes in these young patients are excellent.

Territory and population attributes affect Florida scrub-jay fecundity in fire-adapted ecosystems.

Fecundity, the number of young produced by a breeding pair during a breeding season, is a primary component in evolutionary and ecological theory and applications. Fecundity can be influenced by many environmental factors and requires long-term study due to the range of variation in ecosystem dynamics. Fecundity data often include a large proportion of zeros when many pairs fail to produce any young during a breeding season due to nest failure or when all young die independently after fledging. We conducted color banding and monthly censuses of Florida scrub-jays (Aphelocoma coerulescens) across 31 years, 15 populations, and 761 territories along central Florida's Atlantic coast. We quantified how fecundity (juveniles/pair-year) was influenced by habitat quality, presence/absence of nonbreeders, population density, breeder experience, and rainfall, with a zero-inflated Bayesian hierarchical model including both a Bernoulli (e.g., brood success) and a Poisson (counts of young) submodel, and random effects for year, population, and territory. The results identified the importance of increasing "strong" quality habitat, which was a mid-successional state related to fire frequency and extent, because strong territories, and the proportion of strong territories in the overall population, influenced fecundity of breeding pairs. Populations subject to supplementary feeding also had greater fecundity. Territory size, population density, breeder experience, and rainfall surprisingly had no or small effects. Different mechanisms appeared to cause annual variation in fecundity, as estimates of random effects were not correlated between the success and count submodels. The increased fecundity for pairs with nonbreeders, compared to pairs without, identified empirical research needed to understand how the proportion of low-quality habitats influences population recovery and sustainability, because dispersal into low-quality habitats can drain nonbreeders from strong territories and decrease overall fecundity. We also describe how long-term study resulted in reversals in our understanding because of complications involving habitat quality, sociobiology, and population density.

Mechanistic evidence and exercise interventions: Causal claims, extrapolation, and implementation.

RATIONALE: Exercise interventions and policies are widely prescribed in both sport and healthcare. Research investigating exercise interventions and policies is generally conducted using an Evidence-Based framework, placing an emphasis on evidence gathered from randomised controlled trials (RCTs). AIMS AND OBJECTIVES: To explore the idea that, in addition to the assessment of evidence from RCTs when investigating exercise interventions, mechanistic studies ought to also be assessed and considered. METHODS: This article assesses the rationale supporting the use of RCTs as evidence for exercise interventions, and the use of evidence of mechanisms in establishing efficacy, determining external validity, and tailoring interventions. RESULTS AND CONCLUSIONS: The article argues that evidence from mechanistic studies ought to be considered alongside evidence from RCTs because: as RCTs investigating exercise interventions tend to be of low quality, mechanistic studies ought to be used to reinforce the evidence base; further, evidence from mechanistic studies is highly useful for both questions of extrapolation and implementation. This article argues for this on theoretical grounds, and also draws on a number of case studies.

A comparative physical map reveals the pattern of chromosomal evolution between the turkey (Meleagris gallopavo) and chicken (Gallus gallus) genomes.

BACKGROUND: A robust bacterial artificial chromosome (BAC)-based physical map is essential for many aspects of genomics research, including an understanding of chromosome evolution, high-resolution genome mapping, marker-assisted breeding, positional cloning of genes, and quantitative trait analysis. To facilitate turkey genetics research and better understand avian genome evolution, a BAC-based integrated physical, genetic, and comparative map was developed for this important agricultural species. RESULTS: The turkey genome physical map was constructed based on 74,013 BAC fingerprints (11.9 × coverage) from two independent libraries, and it was integrated with the turkey genetic map and chicken genome sequence using over 41,400 BAC assignments identified by 3,499 overgo hybridization probes along with > 43,000 BAC end sequences. The physical-comparative map consists of 74 BAC contigs, with an average contig size of 13.6 Mb. All but four of the turkey chromosomes were spanned on this map by three or fewer contigs, with 14 chromosomes spanned by a single contig and nine chromosomes spanned by two contigs. This map predicts 20 to 27 major rearrangements distinguishing turkey and chicken chromosomes, despite up to 40 million years of separate evolution between the two species. These data elucidate the chromosomal evolutionary pattern within the Phasianidae that led to the modern turkey and chicken karyotypes. The predominant rearrangement mode involves intra-chromosomal inversions, and there is a clear bias for these to result in centromere locations at or near telomeres in turkey chromosomes, in comparison to interstitial centromeres in the orthologous chicken chromosomes. CONCLUSION: The BAC-based turkey-chicken comparative map provides novel insights into the evolution of avian genomes, a framework for assembly of turkey whole genome shotgun sequencing data, and tools for enhanced genetic improvement of these important agricultural and model species.

Long-term outcome in 42 pediatric liver transplant patients with alpha 1-antitrypsin deficiency: a single-center experience.

INTRODUCTION: We examined the long-term outcome of transplantation for alpha 1-antitrypsin deficiency (A1ATD). METHOD: Data were reviewed on 42 transplants in 35 children with A1ATD over 42 yr and compared with 129 transplants in 116 children with biliary atresia (BA). RESULTS: Over 50% of patients were followed up for >10 yr. A1ATD were older than BA at transplantation, median age, 6.0 vs. 1.0 yr (p < 0.0001), and transplanted earlier in the course of liver failure (total bilirubin, 2.7 mg/dL [1.4-6.9] vs. 9.7 mg/dL [2.9-15.4], p = 0.005). Patient survival was greater in A1ATD than BA: one-yr post-transplant, 82.7% vs. 67.9%; five yr, 76.5% vs. 60.2%; and 10 yr, 76.5% vs. 55.9% (p = 0.03). Death-censored graft survival was similar: one-yr post-transplant, 68.4% vs. 66.2%; five yr, 68.4% vs. 55.8%; and 10 yr, 68.4% vs. 52.5% (p = 0.2). Deaths were from infection, hemorrhage, and graft failure <6 months post-transplant. Patient survival improved at five yr from 33.3% pre-cyclosporine (CSA) (1969-1984) (n = 6) to 76.5% in the CSA era (1985-1994) (n = 17) and 100% with tacrolimus (1995-2006) (n = 12) (p = 0.007). CONCLUSIONS: The age at transplantation and the degree of liver dysfunction were related to the differences in graft and patient survival between A1AT and BA.

Raman spectroscopy enables phenotyping and assessment of nutrition values of plants: a review.

Our civilization has to enhance food production to feed world's expected population of 9.7 billion by 2050. These food demands can be met by implementation of innovative technologies in agriculture. This transformative agricultural concept, also known as digital farming, aims to maximize the crop yield without an increase in the field footprint while simultaneously minimizing environmental impact of farming. There is a growing body of evidence that Raman spectroscopy, a non-invasive, non-destructive, and laser-based analytical approach, can be used to: (i) detect plant diseases, (ii) abiotic stresses, and (iii) enable label-free phenotyping and digital selection of plants in breeding programs. In this review, we critically discuss the most recent reports on the use of Raman spectroscopy for confirmatory identification of plant species and their varieties, as well as Raman-based analysis of the nutrition value of seeds. We show that high selectivity and specificity of Raman makes this technique ideal for optical surveillance of fields, which can be used to improve agriculture around the world. We also discuss potential advances in synergetic use of RS and already established imaging and molecular techniques. This combinatorial approach can be used to reduce associated time and cost, as well as enhance the accuracy of diagnostics of biotic and abiotic stresses.

An Evaluation of the Use of Aggressive Fluid Resuscitation in the Early Treatment of Sepsis Patients.

Introduction Fluid resuscitation is a critical aspect of the sepsis protocol with the usual initial dose being 30 mL per kilogram. Although this dose is well accepted in patients with normal cardiac function, there is some significant variation in clinical practice concerning the optimal fluid resuscitation in septic patients with underlying congestive heart failure (CHF). Many different approaches have been tried to best treat these patients by using lesser volumes of fluid. The purpose of this retrospective study is to attempt to better define optimal fluid resuscitation in congestive heart failure patients and whether standard fluid resuscitation exacerbates CHF in these cases. Methods This was a retrospective study involving patients admitted to the Emergency Department (ED) during the time period of September of 2016 through March of 2019 with a primary diagnosis of sepsis and pre-existing CHF. Data collected from the data warehouse and patient charts included demographics, total amount of fluid received in the ED and outcome data. Evidence of fluid overload (chest X-ray [CXR] evidence, rising B-type natriuretic peptide [BNP], or use of diuretics), was evaluated with respect to in-hospital mortality, white blood cell (WBC) count and comorbidities (chronic obstructive pulmonary disease [COPD], hypertension and coronary artery disease). Results There were 422 patients included in the cohort. Of the 422, 113 (26.8%) patients showed evidence of fluid overload on CXR during hospital stay and received diuretics and therefore considered in the CHF exacerbation group. The patients that experienced CHF exacerbation were significantly older (mean ± SD, 70.9 ± 11.8 years versus 67.4 ± 15.1 years, p=0.014). Patients with exacerbation also received more fluid (median and interquartile range, 3.0, 2:5.5 L versus 2.0, 1:4.3 L, p=0.017). The receiver operating characteristic curve analysis for fluid to predict exacerbation resulted in an area under the curve of 0.59 with a 95% confidence interval (CI) of 0.52 to 0.65, p=0.012. The Youden Index was used to determine an optimal cutoff value of 2.6 L. The percentage of patients in the exacerbation group above the threshold was significantly higher (57.3%) than those without exacerbation (43.3%), p=0.019. Following multivariate analysis, age greater than 60 (odds ratio [OR]: 2.5; CI: 1.4-4.6, p=0.003) and fluid cutoff of 2.6 L (OR: 1.9; CI: 1.2-3.1, p=0.007) were both found to be independent predictors of CHF exacerbation. There was no significant difference in mortality based on the total fluid received in the ED. Conclusion The findings of this study showed that septic patients with pre-existing CHF who received more than 2.6 L of fluid in the ED were 90% more likely to develop symptoms of CHF exacerbation with no evidence of lowering mortality compared to the group that received less than 2.6 L. Our data supports the practice of limiting total fluid resuscitation in CHF to 2.6 L and reconfirms the idea that fluid resuscitation for patients with CHF needs to be individualized.

A Proof-of-Principle Study of Non-invasive Identification of Peanut Genotypes and Nematode Resistance Using Raman Spectroscopy.

Identification of peanut cultivars for distinct phenotypic or genotypic traits whether using visual characterization or laboratory analysis requires substantial expertise, time, and resources. A less subjective and more precise method is needed for identification of peanut germplasm throughout the value chain. In this proof-of-principle study, the accuracy of Raman spectroscopy (RS), a non-invasive, non-destructive technique, in peanut phenotyping and identification is explored. We show that RS can be used for highly accurate peanut phenotyping via surface scans of peanut leaves and the resulting chemometric analysis: On average 94% accuracy in identification of peanut cultivars and breeding lines was achieved. Our results also suggest that RS can be used for highly accurate determination of nematode resistance and susceptibility of those breeding lines and cultivars. Specifically, nematode-resistant peanut cultivars can be identified with 92% accuracy, whereas susceptible breeding lines were identified with 81% accuracy. Finally, RS revealed substantial differences in biochemical composition between resistant and susceptible peanut cultivars. We found that resistant cultivars exhibit substantially higher carotenoid content compared to the susceptible breeding lines. The results of this study show that RS can be used for quick, accurate, and non-invasive identification of genotype, nematode resistance, and nutrient content. Armed with this knowledge, the peanut industry can utilize Raman spectroscopy for expedited breeding to increase yields, nutrition, and maintaining purity levels of cultivars following release.