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The fundamental biology of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid protein (Ncap), its use in diagnostic assays and its potential application as a vaccine component have received considerable attention since the outbreak of the Covid19 pandemic in late 2019. Here we report the scalable expression and purification of soluble, immunologically active, SARS-CoV-2 Ncap in Escherichia coli. Codon-optimised synthetic genes encoding the original Ncap sequence and four common variants with an N-terminal 6His affinity tag (sequence MHHHHHHG) were cloned into an inducible expression vector carrying a regulated bacteriophage T5 synthetic promoter controlled by lac operator binding sites. The constructs were used to express Ncap proteins and protocols developed which allow efficient production of purified Ncap with yields of over 200 mg per litre of culture media. These proteins were deployed in ELISA assays to allow comparison of their responses to human sera. Our results suggest that there was no detectable difference between the 6His-tagged and untagged original Ncap proteins but there may be a slight loss of sensitivity of sera to other Ncap isolates., (© 2024 The Author(s).)

Mammalian zinc metallothionein-3 (Zn 7 MT3) plays an important role in protecting against copper toxicity by scavenging free Cu(II) ions or removing Cu(II) bound to β-amyloid and α-synuclein. While previous studies reported that Zn 7 MT3 reacts with Cu(II) ions to form Cu(I) 4 Zn(II) 4 MT3ox containing two disulfides (ox), the precise localization of the metal ions and disulfides remained unclear. Here, we undertook comprehensive structural characterization of the metal-protein complexes formed by the reaction between Zn 7 MT3 and Cu(II) ions using native ion mobility mass spectrometry (IM-MS). The complex formation mechanism was found to involve the disassembly of Zn 3 S 9 and Zn 4 S 11 clusters from Zn 7 MT3 and reassembly into Cu(I) x Zn(II) y MT3 ox complexes rather than simply Zn(II)-to-Cu(I) exchange. At neutral pH, the β-domain was shown to be capable of binding up to six Cu(I) ions to form Cu(I) 6 Zn(II) 4 MT3 ox , although the most predominant species was the Cu(I) 4 Zn(II) 4 MT3 ox complex. Under acidic conditions, four Zn(II) ions dissociate, but the Cu(I) 4 -thiolate cluster remains stable, highlighting the MT3 role as a Cu(II) scavenger even at lower than the cytosolic pH. IM-derived collision cross sections (CCS) reveal that Cu(I)-to-Zn(II) swap in Zn 7 MT3 with concomitant disulfide formation induces structural compaction and a decrease in conformational heterogeneity. Collision-induced unfolding (CIU) experiments estimated that the native-like folded Cu(I) 4 Zn(II) 4 MT3 ox conformation is more stable than Zn 7 MT3. Native top-down MS demonstrated that the Cu(I) ions are exclusively bound to the β-domain in the Cu(I) 4 Zn(II) 4 MT3 ox complex as well as the two disulfides, serving as a steric constraint for the Cu(I) 4 -thiolate cluster. In conclusion, this study enhances our comprehension of the structure, stability, and dynamics of Cu(I) x Zn(II) y MT3 ox complexes.

Ion mobility-mass spectrometry (IM-MS) unraveled different conformational stability in Zn 4-7 -metallothionein-2. We introduced a new molecular dynamics simulation approach that permitted the exploration of all of the conformational space confirming the experimental data, and revealed that not only the Zn-S bonds but also the α-β domain interactions modulate protein unfolding.

Background: The Metabolic and Bariatric Surgery Accreditation and Quality Improvement Program (MBSAQIP) tracks 30-day outcomes of bariatric patients, but only at accredited centers. Presently, these cases are not broken down by state. Administrative databases can be used to answer some of the questions that are not asked by clinical databases and also to validate those databases. We proposed using the inpatient and outpatient administrative databases in Texas to examine both the numbers and trends of bariatric surgery in Texas over a 5-year period., Methods: The Texas Inpatient Public Use Data File (IPUDF) and the Texas Outpatient Public Data File (OPUDF) were examined for the years 2013-2017. We searched for patients undergoing laparoscopic adjustable gastric banding (LAGB), sleeve gastrectomy (SG), laparoscopic Roux-en-Y gastric bypass (LRYGB) and duodenal switch. Robotic assisted cases were also examined., Results: There were 105,199 bariatric cases performed in Texas from 2013 to 2017. There were 173 centers performing bariatric surgery. The most common operation performed was the sleeve gastrectomy at 73,663 case (70% of total). Gastric bypasses were second at 22,890 cases. During this time period, LAGB almost disappeared; dropping from 2090 cases in 2013 to 115 cases in 2017, with removal of 2097 LAGB in the study period in the OPUDF. During this time period, there was a lower growth rate of the number of SG in the IPUDF with a large increase in SG performed with outpatient status, while LRYGB remained relatively stable., Conclusion: Rates of bariatric surgery in Texas are increasing slowly. The dominant procedure is the SG with a trend towards being done under outpatient status. LAGB has been essentially phased out. There is an increase in SG being performed under 'outpatient' status.

Background: This study aims to measure the effect of guided imagery and hand massage on self-rated wellbeing and pain for palliative care patients., Methods: This study adopted a quasi-experimental one-group pre-test post-test design. The sample consisted of n = 20 adult palliative care patients who received one session of guided imagery and hand massage. Self-reported levels of wellbeing and pain were measured on a scale of 0-10 before and after the intervention. Results were analyzed using a one-tailed sign test in SPSS Software., Results: The intervention elicited a statistically significant improvement in self-reported levels of wellbeing (p = .029) and pain (p = .001). Feedback from participants showed the intervention was helpful and relaxing., Conclusion: The intervention had an immediate positive effect on wellbeing and pain among palliative care patients. Considering the promising results of this pilot study, guided imagery and hand massage should be studied further in the palliative care setting., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Introduction: The American College of Surgeons tracks 30-day outcomes using the Metabolic and Bariatric Surgery Accreditation Quality Initiative Program (MBSAQIP) database. We examined the short-term outcomes of patients that undergo bariatric surgery concomitantly with other operations such as hernia repairs and cholecystectomy to determine the safety of this practice., Methods: The MBSAQIP Participant Use Data File for 2015-2017 was examined for differences in primary bariatric operations vs concomitant procedures (CP). We looked for concurrent CPT codes for laparoscopic cholecystectomy (LC) and hernia repairs (ventral, epigastric, incisional, and inguinal). p was significant at < 0.05., Results: There were 464,674 cases, of which 15,614 had CP. For both LRYGB+LC and SG+LC, there were increased operative times and length of stay. There were statistically significant higher rates of readmission, reintervention, and reoperation for SG+LC vs SG alone, as well as for LRYGB+hernia and SG+hernia. There was a higher risk of death (p < 0.001) in LRYGB+hernia patients. Also, LRYGB+hernia patients had statistically significant increases in unplanned admission to the intensive care unit and pulmonary embolus. SG+hernia patients had a higher rate of ventilation > 48 h, unplanned admission to the ICU, pulmonary embolism, deep vein thrombosis, and readmission, reintervention, and reoperation., Conclusions: There is a statistically higher rate of complications with concomitant procedures in the MBSAQIP database. Length of stay and operative times are increased in concomitant operations as are readmissions, reinterventions, and reoperations. These findings would indicate that additional procedures at the time of bariatric surgery should be deferred if possible.

Background: Metabolic and bariatric surgery (MBS) has been shown to improve medical problems; however, there are known arrhythmias that can occur after MBS (i.e., sick sinus syndrome [SSS] and sinus bradyarrhythmias). While the literature in this area contains case reports, there is a lack of published data on a state or national level. We used a large state administrative database to evaluate the occurrence of cardiac arrhythmias after MBS., Methods: We studied the years 2016 to 2018 using the Texas Inpatient Public Use Data File. Inclusion criteria were patients who had a pacemaker installed and were ≥ 18 years. Quantitative variables were described using mean and standard deviation. Categorical variables were described using frequency and proportion. The student's t-test and chi-squared test were used to assess the differences across pacemaker installation., Results: There were a total of 79,807 (10.2%) who had a history of MBS and 31,072 (4%) patients who underwent pacemaker insertion, respectively. After excluding all patients < 18 years, the prevalence of pacemakers installed in patients with prior bariatric surgery was 0.8% (n = 257/30,823) or about 8 in every 1000 patients. Of note, bariatric patients who had a pacemaker placed were younger than non-bariatric patients (P < 0.001). The most common reason for pacemaker placement was SSS (51.5%), followed by atrioventricular block (13.1%), and then bradycardia at 8.5%. The most common arrhythmia overall was bradycardia., Conclusions: Eight out of every 1000 patients with a pacemaker installed in the study period had a history of MBS. The most common arrhythmia was bradycardia and the most common reason for pacemaker placement was sick sinus syndrome. These results do not indicate causality but may demonstrate an association between MBS and arrhythmias. Bariatric patients undergo pacemaker placement at a younger age. The relationship between bariatric surgery and cardiac arrhythmias warrants further study., Competing Interests: Conflicts of Interest: The authors declare no conflict of interest., (© 2020 by JSLS, Journal of the Society of Laparoscopic & Robotic Surgeons.)

Background : The sleeve gastrectomy (SG) can be associated with postoperative gastroesophageal reflux and when a hiatal hernia (HH) is present, it should be fixed. Earlier studies have shown that 20% of SG have a concomitant hiatal hernia repair (SG+HHR). The aim of this project is to determine the rate of SG+HHR in a large state administrative database. Methods: The Texas Inpatient Public Use Data File (IPUDF) and Outpatient Public Use Data File (OPUDF) for the years 2013-2017 were examined for patients that underwent SG+HHR at the same time. Patient demographics, diagnosis, and charge data were also examined. A t-test was performed between groups and P was considered significant at < 0.05. Results: In the OPUDF, there were 6,193 (33.7%) patients who underwent SG+HHR out of 18,403 patients who underwent SG. Mean charges were $94,741 [standard deviation (SD) = $87,284]. Length of stay (LOS) was 2.1 (SD = 3.5) vs 2.3 days (SD = 3.3) with a shorter stay for SG+HHR vs SG alone (P < 0.001). In the IPUDF, there were 11,536 (21.1%) patients who underwent SG+HHR out of 54,545 patients who underwent SG. Mean charges were $69,006 (SD = $46,365). LOS was 1.59 days (SD = 3.7) for SG+HHR vs 1.63 days (SD = 1.6) for SG (P = .043). The rate of SG+HHR increased over the study period. Conclusions: SG+HHR is common in both the outpatient and inpatient setting. There is a yearly trend of increasing rates of SG+HHR., Competing Interests: Conflicts of Interest: The authors declare no conflict of interest., (© 2020 by JSLS, Journal of the Society of Laparoscopic & Robotic Surgeons.)

Background: Many bariatric surgeons test the anastomosis and staple lines with some sort of provocative test. This can take the form of an air leak test with a nasogastric tube with methylene blue dye or with an endoscopy. The State Department of Health Statistics in Texas tracks outcomes using the Texas Public Use Data File (PUDF)., Methods: We queried the Texas Inpatient and Outpatient PUDFs for 2013 to 2017 to examine the number of bariatric surgeries with endoscopy performed at the same time. We used the International Classification of Diseases Clinical Modification Version 9 (ICD-9-CM) and ICD-10 procedure codes and Current Procedural Terminology for Sleeve Gastrectomy (SG) and laparoscopic Roux-en-Y gastric bypass (LRYGB) and endoscopy, and the ICD-9-CM and ICD-10 diagnosis codes for morbid obesity., Results: There were 74,075 SG reported in the Texas Inpatient and Outpatient PUDF for the years 2013-2017. Of the SG performed, 5,521 (7.4%) had an intraoperative endoscopy. For the 19,192 LRYGB reported, 1640 (8.6%) underwent LRYGB + endoscopy. This was broken down by SG only vs SG + endoscopy and LRYGB only vs LRYGB + endoscopy. Overall, SG + endoscopy had a significantly shorter length of stay (LOS) vs LRYGB + endoscopy at 1.74 d vs 2.34 d ( P < .001) and a significantly less cost of $71,685 vs $91,093 ( P < .001)., Conclusions: A small percentage of SG and LRYGB patients underwent endoscopy for provocative testing over the study period. Provocative testing with endoscopy costs more for SG and LRYGB and was associated with a shorter LOS., Competing Interests: Conflicts of Interest: The authors have no conflicts of interest directly relevant to the content of this article., (© 2020 by JSLS, Journal of the Society of Laparoscopic & Robotic Surgeons.)

Background: The Metabolic and Bariatric Surgery Accreditation and Quality Improvement Program (MBSAQIP) database tracks patients, techniques, and outcomes for 30 days. The overwhelming majority of cases reported are performed using a laparoscopic technique. Bariatric surgeons rarely have to convert from laparoscopy to open surgery., Objectives: We examined the MBSAQIP to determine the characteristics of patients who underwent conversion and evaluated their short-term outcomes., Settings: University program in the United States and nationwide clinical database., Methods: The MBSAQIP Public Use File for 2017 was examined for primary bariatric operations. We identified patients who underwent a sleeve gastrectomy or gastric bypass using a minimally invasive technique. We identified patients who underwent conversion to another operative technique or were converted to open surgery and analyzed preoperative characteristics and postoperative complication rates. Relative risks (RR) were calculated for complications. P value was significant at < .05., Results: There were 186,962 patients in the entire cohort. Six hundred nine patients underwent conversion from the original surgical approach to either open surgery (n = 457) or to another technique (n = 152). Patients with preoperative oxygen dependency, poor functional status, previous foregut/obesity surgery, preoperative renal insufficiency, and anticoagulation were more likely to undergo conversion. Patients who underwent conversion to the open approach had longer operative times (191 versus 86.6 min [P < .001]) and longer time to discharge (6.2 versus 1.6 d [P < .001]). The RR of death was 18.2 (95% confidence interval 8.7-37.6, P < .001) for procedures converted to open. The RR of sepsis was 10.1 (95% confidence interval 4.2-24.2, P < .001) and the RR for all complications was increased throughout for patients undergoing conversion., Conclusions: Patients in the MBSAQIP database that undergo conversion to the open surgical approach are at a greatly increased risk for death and complications., (Copyright © 2020 American Society for Bariatric Surgery. Published by Elsevier Inc. All rights reserved.)

Background: Access to bariatric care varies across regions, ethnic, and racial groups. Some of these variations may be due to insurance status or socioeconomic status. There are also regional and state variations in access to metabolic and bariatric surgery (MBS). The Texas Inpatient Public Use Data File (IPUDF) and Texas Outpatient Public Use Data File is a state-mandated database that collects information on demographics, procedures, diagnoses, and cost on almost all admissions in Texas. We used them to examine racial disparities in MBS over a 5-y period., Methods: The IPUDF and Texas Outpatient Public Use Data File were examined from the years 2013 through, 2017. We included all patients undergoing a laparoscopic Roux-en-Y gastric bypass and sleeve gastrectomy and examined the demographics of these patients. Race and ethnicity are reported separately. We used U.S. Census Bureau statistics and the Texas Department of State Health Services statistics to determine the crude (unadjusted) and adjusted procedure rates of patients undergoing MBS., Results: In the IUPUDF, the crude unadjusted procedure rate for blacks undergoing MBS was 7.29 per 10,000 population followed by 6.85 per 10,000 for non-Hispanic whites. Hispanics had the lowest rate at 3.20 per 10,000. When adjusted for sex, obesity, age, and race, blacks still had a higher rate of access followed by whites and then Hispanics., Conclusions: There are disparities to access for bariatric surgery in Texas. Blacks have the greatest access followed by whites. Hispanics have the lowest procedure rate per population., Competing Interests: Conflicts of Interest: The authors declare no conflict of interest., (© 2020 by JSLS, Journal of the Society of Laparoscopic & Robotic Surgeons.)

Background: The most common bariatric operation in the United States is sleeve gastrectomy. The second and third most common bariatric operations are gastric bypass and revisional bariatric surgery, respectively., Objective: The objective of the study was to assess the differences between laparoscopic revisional weight loss surgery (LRWLS) and robotic revisional weight loss surgery (RRWLS)., Setting: University hospital, United States., Methods: Data were extracted from the Metabolic and Bariatric Surgery Accreditation and Quality Improvement Program database spanning 2015 to 2016 to look at demographic characteristics, operative time, co-morbidities, and length of stay. Using the specified Current Procedural Terminology codes, patients who underwent bariatric procedures and required a revisional procedure were identified., Results: A total of 354,865 patients were included in this study; 37,917 (11.9%) patients required revision after undergoing a bariatric procedure. Of these revisions, 94.9% (n = 35,988) were LRWLS, and 5.1% (n = 1929) were RRWLS. There were no differences in patient characteristics between the LRWLS and RRWLS groups. There was a significant difference between the RRWLS and the LRWLS groups in operative time, with the RRWLS group taking 167 minutes and the LRWLS group taking 103 minutes (P < .001). There was a statistically significant increase in length of stay for RRWLS, 2.3 days versus 1.7 for LRWLS (P < .005). In terms of postoperative complications, there were no significant differences between the 2 groups., Conclusions: RRWLS is as safe as LRWLS in the Metabolic and Bariatric Surgery Accreditation and Quality Improvement Program database. There is an increase in operative times and length of stay for robotic cases., (Copyright © 2019 American Society for Bariatric Surgery. Published by Elsevier Inc. All rights reserved.)