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Summary: Human pluripotent stem cell (hPSC)-derived tissues can be used to model diseases in cell types that are challenging to harvest and study at-scale, such as neutrophils. Neutrophil dysregulation, specifically neutrophil extracellular trap (NET) formation, plays a critical role in the prognosis and progression of multiple diseases, including COVID-19. While hPSCs can generate limitless neutrophils (iNeutrophils) to study these processes, current differentiation protocols generate heterogeneous cultures of granulocytes and precursors. Here, we describe a method to improve iNeutrophil differentiations through the deletion of GATA1. GATA1 knockout (KO) iNeutrophils are nearly identical to primary neutrophils in form and function. Unlike wild-type iNeutrophils, GATA1 KO iNeutrophils generate NETs in response to the physiologic stimulant lipopolysaccharide, suggesting they are a more accurate model when performing NET inhibitor screens. Furthermore, through deletion of CYBB, we demonstrate that GATA1 KO iNeutrophils are a powerful tool in determining involvement of a given protein in NET formation.

Restoration of glenoid bony integrity is critical to minimizing the risk of recurrence and re-creating normal kinematics in the setting of anterior glenohumeral instability. We present an arthroscopic suture anchor–based technique for treating large bony Bankart fractures in which the fragment is secured to the intact glenoid using mattress sutures placed through the bony fragment and augmented with soft-tissue repair proximal and distal to the bony lesion. This straightforward technique has led to excellent fragment reduction and good outcomes in our experience.

Objective: The objective of this work was to describe an efficient and sustainable outreach model in a resource-constrained environment, with a multifaceted approach focusing on national policy change, telemedicine, injury prevention, education and treatment of burns., Summary Background Data: Burn injury constitutes a significant portion of morbidity and mortality worldwide, particularly in children, and in low- and middle-income countries., Methods: We reviewed the impact of ten-years of a burn outreach program. Our focus was on clinical data on burn care within one region of Ukraine. We assessed knowledge of burn prevention/first aid utilizing a large survey, analyzed clinical data from our outreach clinic and telemedicine program, and analyzed data within a newly created burn repository within Ukraine., Results: A national burn prevention policy has been implemented through our efforts along with a burn prevention program. Educational efforts have led to improvements of major complication rates (wound infection [7% vs. 16%], pneumonia [2.4% vs. 0.3%], sepsis [1.6% vs. 0.6%], UTI [2% vs. 0.6%], and cellulitis [11% vs. 3.4%]), respiratory support of acutely ill patients [1.3% vs. 0.4%], and blood transfusion triggers., Conclusions: Broadly, our model could be an example of building sustainable outreach programs in resource-constrained environments. Through collaboration with local healthcare providers, we have developed and implemented an outreach program in a resource-constrained environment., (Copyright © 2021 Elsevier Ltd and ISBI. All rights reserved.)

The successful commercialization of cell therapies requires thorough planning and consideration of product quality, cost and scale of the manufacturing process. The implementation of automation can be central to a robust and reproducible manufacturing process at industrialized scales. There have been a number of wash-and-concentrate devices developed for cell manufacturing. These technologies have arisen from transfusion medicine, hematopoietic stem cell and biologics manufacturing where operating mechanisms are distinct from manual centrifugation. This review describes the historical origin and fundamental technologies underlying each currently available wash-and-concentrate device as well as their relative advantages and disadvantages in cell therapy applications. Understanding the specific attributes and limitations of these technologies is essential to optimizing cell therapy manufacturing., (Copyright © 2021 International Society for Cell & Gene Therapy. Published by Elsevier Inc. All rights reserved.)

Background: Cost-analysis research can influence healthcare policies and practices. There is inherent bias depending on the chosen cost perspective (hospital, third-party payer, societal), and conclusions can change based on the perspective used. These perspectives may or may not be well declared or justified when performing cost-analysis research. The goal of this study was to perform a literature review of cost-analysis research in the Plastic Surgery literature to determine the prevalence of studies declaring and justifying their perspective, and to inform the reader on why such declarations are important in understanding potential bias., Methods: A systematic review was completed to retrieve cost-utility and cost-effectiveness research within the scope of Plastic Surgery. The search was limited to English-language studies in North America and Europe published between 2006 and 2016. Articles were selected using predefined data fields and specific inclusion criteria., Results: A total of 2304 abstracts were identified, of which 47 met inclusion criteria. Seventy-two percent of studies (n = 34) declared a cost perspective. Of the studies that identified a cost perspective, 32% incorrectly identified the cost perspective. Only 49% of all studies (n = 23) both accurately declared and justified their chosen perspective., Conclusions: Only half of studies correctly declare their cost perspective and justify why the perspective was chosen. Not doing so potentially hides bias from the reader. Future efforts when performing cost-analysis studies should require a clear declaration and justification of the cost perspective taken. A table of our recommendations for reporting cost perspective is provided., (Copyright © 2017 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.)

Children's misuse of harmful legal products (HLPs), including inhaling or ingesting everyday household products, prescription drugs, and over-the-counter drugs, constitutes a serious health problem for American society. This article presents a community prevention model (CPM) focusing on this problem among pre and early adolescents. The model, consisting of a community mobilization strategy and environmental strategies targeting homes, schools, and retail outlets, is designed to increase community readiness and reduce the availability of HLPs, which is hypothesized to reduce HLPs use among children. The CPM is being tested in Alaskan rural communities as part of an inprogress eight-year National Institute on Drug Abuse randomized-controlled trial. This paper presents the CPM conceptual framework, describes the model, and highlights community participation, challenges, and lessons learned from implementation of the model over a 21-month period., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The proximal ligand of thiolate-coordinated heme proteins is crucial for the activation of the oxygen molecule and hydroxylation of substrates. In nitric oxide synthases (NOSs), the heme axial cysteine ligand forms a hydrogen bond to the side chain indole nitrogen of a tryptophan residue. Resonance Raman spectroscopy was used to probe W56F and W56Y variants of the NOS of Staphylococcus aureus (saNOS) and the analogous W180 variants of the endothelial NOS oxygenase domain (eNOSox). We show that the variants displayed lower nu(Fe-NO) and nu(Fe-CO) frequencies indicating that these mutations increased the electron density on the axial cysteine in their Fe(III)NO and Fe(II)CO complexes. We also show by UV-visible spectroscopy that the Fe(II)CO complexes of the variants displayed a red-shifted Soret optical transition in addition to the lower nu(Fe-CO) thus establishing that these properties are sensitive indicators of the modulation of the basicity of the axial cysteine. We infer, based on its spectroscopic properties, that ferrous eNOSox W180Y saturated with l-arginine and tetrahydrobiopterin forms a tyrosine-cysteine hydrogen bond when bound to CO. Evidence for such a hydrogen bond was not obtained for the Fe(III)NO protein nor for the analogous saNOS variant. These mutations reveal interesting differences in the response of NOS isotypes to analogous mutations at conserved residues and clearly show that the heme-Fe to cysteine sigma bond is modulated by the Cys-Trp hydrogen bond in NOSs. These studies serve as a basis to gain information on the role played by this hydrogen bond in oxygen activation in this class of enzymes.

The acoustic startle reflex can be modulated by positive and negative emotion. There is evidence that this modulation can be influenced by voluntary attempts to regulate emotion, and that startle modulation during emotion regulation is more reflective of changes in arousal than valence. However, whether valence and arousal play similar roles in emotion regulation across different psychophysiological indices is unclear. The goal of this study was to characterize further the relative contributions of valence and arousal to changes in psychophysiological responsiveness during voluntary emotion regulation, using multiple psychophysiological measures including eyeblink startle, skin conductance, and heart rate. We studied 10 healthy adults, and found that voluntary attempts to down-regulate positive and negative emotion resulted in decreased eyeblink startle magnitude, skin conductance responses, and heart rate, relative to attempts to up-regulate emotion. These findings indicate that the volitional regulation of emotion had systematic effects on psychophysiological parameters which were similar for positive and negative emotion, suggesting that psychophysiological responsiveness during emotion regulation is more strongly influenced by the modulation of arousal than by the valence of the regulated emotion.

Objective: Surgery is the cornerstone of management in ovarian cancer. However, in high-risk and elderly patients there is a tendency for less aggressive surgery upfront. The aim of this study was to review cytoreductive surgery, with focus on complications and outcomes in patients with multiple surgical risk factors., Study Design: Charts of patients with ovarian cancer from 1998 to 2002 were retrospectively reviewed., Results: One hundred and forty patients were treated for ovarian cancer. Sixty-three patients (45%) were elderly (> or =65 years), and 69 patients (49%) had comorbidities. Optimal debulking (< or =1 cm) was achieved in 123 patients (88%). There was no significant relation between complications and type of procedure, elderly age, comorbidities, or transfusions. Optimally debulked patients had a significantly longer survival than patients with suboptimal debulking (P < .001)., Conclusion: Aggressive optimal cytoreduction can be achieved in the majority of patients with multiple surgical risk factors and is associated with a low complication rate.

Objective: Approximately 50% of patients with ovarian cancer who have normal CA 125 levels at the completion of therapy have persistent disease. In an effort to improve the ability to detect small volume disease, we have evaluated the usefulness of N-acetylglucosamine:beta1,4-galactosyltransferase as a potential biomarker for the detection of subclinical disease after the completion of primary therapy for ovarian cancer., Study Design: The sera of 33 patients with stage IIIC epithelial ovarian cancer in complete clinical remission after chemotherapy (CA 125 <35 units/mL and negative computed tomography scan) who underwent second-look surgery were examined for N-acetylglucosamine:beta1,4-galactosyltransferase activity. The values were determined from sera that had been obtained before primary cytoreductive operation and before second-look surgery after the completion of platinum-based chemotherapy. Determinations of the levels of CA 125 were performed with the Bayer Immuno ITM CA-125 II assay. N-acetylglucosamine:beta1,4-galactosyltransferase activity was determined by measuring the transfer of galactose from uridine diphosphate- carbon 14-labeled galactose to the terminal N-acetylglucosamine residue of a very well-defined synthetic acceptor, N-acetylglucosamine:beta1,6GalNAc(alpha)-o-benzyl, which is a portion of the core structure of mucin glycoproteins. The cutoff value of N-acetylglucosamine:beta1,4-galactosyltransferase was determined to be 22,000 counts/min, based on the analysis of 25 healthy control subjects. Correlation between serum CA 125 and N-acetylglucosamine:beta1,4-galactosyltransferase levels was determined with the use of the Pearson correlation coefficient. The ability of galactosyltransferase to identify small volume disease correctly was also evaluated., Results: There was a significant correlation between serum CA 125 and N -acetylglucosamine:beta1,4-galactosyltransferase levels before the operation (r = 0.57; P =.03) but not before second-look surgery (r = 0.10; P =.57). Thirteen patients (39.4%) had residual disease at second-look surgery. Elevated N-acetylglucosamine:beta-1,4galactosyltransferase activity >22,000 cpm correctly identified 10 of these patients (76.9%). The sensitivity, specificity, and positive and negative predictive values of N-acetylglucosamine:beta1,4-galactosyltransferase activity (>22,000 counts/min) for the prediction of residual disease at second-look surgery were 77%, 45%, 48%, and 77%, respectively., Conclusion: Our comparative study of serum CA 125 and N -acetylglucosamine:beta1,4-galactosyltransferase levels showed a significant correlation between the two tumor markers before the beginning of ovarian cancer therapy. This correlation disappeared before second-look surgery because 60% of patients with normal serum CA 125 and N-acetylglucosamine:beta1,4-galactosyltransferase levels. CA 125 antigen appears to be inferior to N -acetylglucosamine:beta1,4-galactosyltransferase in the detection of small-volume residual disease. N-acetylglucosamine:beta1,4-galactosyltransferase may be useful as a biomarker in the monitoring of patients with ovarian cancer when the serum CA 125 level is normal. These findings require confirmation in larger studies.

A retrospective review of our institute's tumor registry from January 1972 to January 1996 revealed 24 patients with a diagnosis of primary soft tissue sarcoma of the hand, from a total of 570 extremity soft tissue sarcomas (4%). The most frequent histologic type was malignant fibrous histiocytoma, which occurred in 9 (38%) of the 24 patients. The second most common histologic type was epithelioid sarcoma, which occurred in 6 (25%) patients. There was a statistically significant difference in the rate of local recurrence based on the type of treatment in which amputation was superior to the other forms of treatment. There was no statistically significant difference in the rate of distant failures between treatment groups. The estimated cumulative 5- and 10-year overall survival rates for all patients were 59% and 53%, respectively. Stage II patients had estimated cumulative 5- and 10-year survival rates of 68% and 59%, respectively. Stage III patients had a cumulative 5-year survival rate of 20%. Factors that were statistically significant in predicting survival were the size of the primary tumor, with tumors smaller than 5 cm having a better prognosis, and stage of the tumor at presentation, with stage I and II tumors having the highest survival rate. In selected patients with a primary hand sarcoma, aggressive limb-sparing surgery with adjuvant therapy offered equivalent survival compared with amputation.

As a component of a staged laparotomy for trauma, perihepatic packing can be lifesaving in the patient with exsanguinating liver injury who, due to hypothermia and coagulopathy, is unable to tolerate a more extensive procedure. However, if intra-abdominal pressure increases, the manoeuvre has been reported to compromise cardiopulmonary stability. Patients who suffer the adverse consequences of intra-abdominal hypertension are commonly managed with a loose temporary closure, frequently using an artificial material to bridge the skin defect across the incision. A case is reported where these two seemingly contradictory options were combined to achieve a successful outcome.

The synthetic retinobenzoic acid RE-80 was evaluated for its potential as an inductor of tumor cell differentiation and as a chemopreventive agent. A minimally toxic dose of RE-80 in vitro produced morphologic changes typical differentiation in epidermal tumor cell colonies. Indirect immunofluorescence indicated induction of a differentiation-associated keratin of internal stratified epithelia, K13, and inhibition of the differentiation-associated epidermal keratin K1. Cultured cells were skin-grafted to athymic nu/nu mice to evaluate RE-80 effects on early stage malignant progression in vivo. When tumors had grown to 3 to 4 mm in diameter, mice were treated by intraperitoneal injection with RE-80 (67 micrograms, 170 mmol, i.p., two times per week) or vehicle (100 microliters 20% ethanol). Papillomas (benign) and moderately differentiated squamous cell carcinomas were reduced in volume about 4-fold by RE-80 treatment. Larger, poorly differentiated squamous cell carcinomas were unaffected. RE-80 resulted in a lower proportion of proliferative cells (detectable by bromodeoxyuridine incorporation) and a higher proportion of moderately to well differentiated tumors after 40 days of treatment compared with control tumors, which were mainly poorly differentiated squamous cell carcinomas. K13 induction in vitro was correlated with response to retinoid in vivo. Induction of differentiation may be mechanism of the response to RE-80 in vivo since carcinoma cells expressing K13 did not incorporate bromodeoxyuridine and were on a terminal pathway.

In 53 patients with histologically proven adrenal cortical carcinomas the most common metastatic sites, at the time of diagnosis, were the liver, lymph nodes, bone and lungs, and during the course of the disease, the lungs. At the time of diagnosis six (11%) of our patients were in Stage S1, seven (13%) in S2, 11 (21%) in S3 and 29 (55%) in S4. Histologic grading (G) in 43 patients showed G1 grade in 11 (26%), G2 in seven (16%) and G3 in 25 patients (58%). Median survival in months (estimated 5-year survival rate) for the various stages were: S1 46 (33%), S2 84 (57%), S3 eight (18%) and S4 seven (7% at 2 years) (P = 0.01). Median survival in months (estimated 5-year survival rate) for the various grades were: G1 46 (46%), G2 24 (43%), G3 five (4% at 2 years) (P = 0.003). In this series, tumor stage (P = 0.01) and grade (P = 0.003) were predictors of survival and the two were closely correlated (P = 0.02).

One hundred three consecutive patients undergoing 106 thoracotomies for primary lung carcinoma were reviewed to determine factors associated with the development of postoperative pulmonary complications. Pulmonary complications occurred in 40 of 104 (39 percent) patients. Minor complications occurred in 17 of 104 (16 percent) patients and major in 23 of 104 (22 percent). There were six deaths in the entire series of 103 patients (6 percent), two of which were directly caused by a pulmonary complication and one where it was a contributing factor. Extended surgical resections were associated with an increased risk of complications. Pulmonary complications occurred in 9 of 11 (82 percent) patients undergoing extended resections involving chest wall resection. The use of neoadjuvant chemotherapy also was associated with an increase in the rate of major complications. Poor nutritional status as measured by a history of weight loss and preoperative serum albumin levels also was associated with an increased risk of any pulmonary complication. Cardiac complications were significantly increased in the group of patients having pulmonary complications. Pulmonary complications continue to present a major source of morbidity and mortality for patients undergoing thoracotomy for lung carcinoma. Determination of factors associated with increased risk is important in order to identify patients who might be predisposed to the development of these complications.

A field trial of artificial insemination in horses with transproted, chilled semen was conducted using a specially designed container which permitted a controlled, slow initial rate of cooling (-0.3 degrees C/min) and maintenance of a final temperature of 4 degrees -6 degrees C for more than 36 hrs. Forty-six mares in 23 states were inseminated with semen from three German Warmblood stallions standing at stud in Hamilton, Massachusetts. A third-cycle conception rate of 91% was obtained.