Moxonidine Increases Uptake of Oxidised Low-Density Lipoprotein in Cultured Vascular Smooth Muscle Cells and Inhibits Atherosclerosis in Apolipoprotein E-Deficient Mice.

This study aimed to investigate the effect of the sympatholytic drug moxonidine on atherosclerosis. The effects of moxonidine on oxidised low-density lipoprotein (LDL) uptake, inflammatory gene expression and cellular migration were investigated in vitro in cultured vascular smooth muscle cells (VSMCs). The effect of moxonidine on atherosclerosis was measured by examining aortic arch Sudan IV staining and quantifying the intima-to-media ratio of the left common carotid artery in apolipoprotein E-deficient (ApoE-/-) mice infused with angiotensin II. The levels of circulating lipid hydroperoxides in mouse plasma were measured by ferrous oxidation-xylenol orange assay. Moxonidine administration increased oxidised LDL uptake by VSMCs via activation of α2 adrenoceptors. Moxonidine increased the expression of LDL receptors and the lipid efflux transporter ABCG1. Moxonidine inhibited mRNA expression of inflammatory genes and increased VSMC migration. Moxonidine administration to ApoE-/- mice (18 mg/kg/day) decreased atherosclerosis formation in the aortic arch and left common carotid artery, associated with increased plasma lipid hydroperoxide levels. In conclusion, moxonidine inhibited atherosclerosis in ApoE-/- mice, which was accompanied by an increase in oxidised LDL uptake by VSMCs, VSMC migration, ABCG1 expression in VSMCs and lipid hydroperoxide levels in the plasma.

The effects of hyperbaric oxygen on MRI findings in rheumatoid arthritis: A pilot study.

Background: Rheumatoid arthritis is a debilitating and destructive disease for which limited therapeutic options exist. Objective: This report summarizes serial magnetic resonance imaging (MRI) findings from nine study participants treated with hyperbaric oxygen (HBO2) therapy and expands upon an earlier pilot study that showed improvement in disease activity and joint pain as determined by multiple, validated clinical measures. Methods: Rheumatoid arthritis patients received 30 hyperbaric oxygen treatments over six to 10 weeks. MRI with and without contrast was completed at baseline, and at three- and six-month intervals following initiation of HBO2 therapy. Ratings were based on Outcome Measures in Rheumatology Clinical Trials (OMERACT) Rheumatoid Arthritis Magnetic Resonance Imaging Score (RAMRIS) criteria, the standard method for quantification of inflammation and damage by MRI in RA trials. Results: Using RAMRIS criteria, nine of nine patients demonstrated no radiologic progression of erosions, synovitis, or bone marrow edema at three- and six-month scans. Conclusion: Our findings suggest that HBO2 therapy may be useful as an adjunctive or alternative treatment to disease-modifying drugs for rheumatoid arthritis.

Knock down of TIMP-2 by siRNA and CRISPR/Cas9 mediates diverse cellular reprogramming of metastasis and chemosensitivity in ovarian cancer.

BACKGROUND: The endogenous tissue inhibitor of metalloproteinase-2 (TIMP-2), through its homeostatic action on certain metalloproteinases, plays a vital role in remodelling extracellular matrix (ECM) to facilitate cancer progression. This study investigated the role of TIMP-2 in an ovarian cancer cell line in which the expression of TIMP-2 was reduced by either siRNA or CRISPR/Cas9. METHODS: OVCAR5 cells were transiently and stably transfected with either single or pooled TIMP-2 siRNAs (T2-KD cells) or by CRISPR/Cas9 under the influence of two distinct guide RNAs (gRNA1 and gRNA2 cell lines). The expression of different genes was analysed at the mRNA level by quantitative real time PCR (qRT-PCR) and at the protein level by immunofluorescence (IF) and western blot. Proliferation of cells was investigated by 5-Ethynyl-2'-deoxyuridine (EdU) assay or staining with Ki67. Cell migration/invasion was determined by xCELLigence. Cell growth in vitro was determined by 3D spheroid cultures and in vivo by a mouse xenograft model. RESULTS: Approximately 70-90% knock down of TIMP-2 expression were confirmed in T2-KD, gRNA1 and gRNA2 OVCAR5 ovarian cancer cells at the protein level. T2-KD, gRNA1 and gRNA2 cells exhibited a significant downregulation of MMP-2 expression, but concurrently a significant upregulation in the expression of membrane bound MMP-14 compared to control and parental cells. Enhanced proliferation and invasion were exhibited in all TIMP-2 knocked down cells but differences in sensitivity to paclitaxel (PTX) treatment were observed, with T2-KD cells and gRNA2 cell line being sensitive, while the gRNA1 cell line was resistant to PTX treatment. In addition, significant differences in the growth of gRNA1 and gRNA2 cell lines were observed in in vitro 3D cultures as well as in an in vivo mouse xenograft model. CONCLUSIONS: Our results suggest that the inhibition of TIMP-2 by siRNA and CRISPR/Cas-9 modulate the expression of MMP-2 and MMP-14 and reprogram ovarian cancer cells to facilitate proliferation and invasion. Distinct disparities in in vitro chemosensitivity and growth in 3D culture, and differences in tumour burden and invasion to proximal organs in a mouse model imply that selective suppression of TIMP-2 expression by siRNA or CRISPR/Cas-9 alters important aspects of metastasis and chemosensitivity in ovarian cancer.

Modification of hurling helmets causing penetrating upper limb injury: A case series demonstrating opportunity for improved player safety.

BACKGROUND: The games of hurling and camogie involve fast ball movement and a substantial physical contact with opposing players. The wearing of protective helmets with faceguards became obligatory 10 years ago and resulted in a significant reduction of craniofacial injuries. However since then a trend has emerged of players modifying helmets to improve comfort and vision. As officials are not yet required to perform pre-match checks, many players enter competition with defective protective equipment which poses a risk to both themselves and others. METHODS: We present 4 cases of penetrating upper limb injury in hurlers sustained as a result of helmet faceguards. These injuries were sustained during competitive play and the affected patients presented with portions of the faceguards embedded in their wounds. Each patient required admission to hospital and exploration of their wounds under general anaesthetic. CONCLUSIONS: This case series demonstrates the dangers of modification and damage to protective helmet faceguards used in hurling and camogie. We encourage helmet manufacturers to re-engage with players to ensure that current safety equipment meets the functional demands of the modern player while adhering to approved manufacturing standards. Mandatory helmet checks before competitive games would reduce the likelihood of penetrating upper limb injury to players.

Patients Older 65 Years With Early Breast Cancer Prefer Intraoperative Radiation as a Locoregional Treatment Choice.

BACKGROUND: Patients 65 years old or older with early endocrine-responsive breast cancer have many treatment options, including no radiation. This study aimed to evaluate treatment preference when intraoperative radiation therapy (IORT) is offered in this population. METHODS: The study reviewed patients 65 years old or older with a diagnosis of early-stage endocrine-responsive breast cancer in 2016-2019 at a single hospital in a large integrated health care system. Electronic medical records of multidisciplinary breast tumor board discussion, treatment options documented by the treatment team, and final treatment offered were reviewed. Variables including age at biopsy, language, endocrine treatment, and comorbidities were collected. Regression analysis was used to evaluate for variables associated with patients' choice regarding radiation treatment. RESULTS: The institutional IORT guidelines were met by 63 patients in the described age group who had a documented offer of all radiation treatment options. The median age of the patients was 70 years (interquartile range 63-77 years). Overall, 74.6% of the patients chose IORT, and 14.3% opted for whole-breast irradiation. Only 4.8% chose to omit radiation after breast-conserving surgery, and 6.3% chose mastectomy. The patients who chose IORT were more likely to receive endocrine treatment (odds ratio 3.70; p = 0.03). Age, race, language, and comorbidities were not associated with preference for IORT (p < 0.05). CONCLUSIONS: Patients 65 years old or older with early-stage endocrine-responsive breast cancer preferred to have IORT despite counsel about the lack of survival benefit. This study suggests that local cancer control with the convenient radiation delivery method is important to the described patient population.

Virtual Reality Warm-up Before Robot-assisted Surgery: A Randomized Controlled Trial.

TRIAL DESIGN: This was a randomized controlled trial. BACKGROUND: Intraoperative errors correlate with surgeon skill and skill declines with intervals of inactivity. The goals of this research were to identify the optimal virtual reality (VR) warm-up curriculum to prime a surgeon's technical skill and validate benefit in the operating room. MATERIALS AND METHODS: Surgeons were randomized to receive six trial sessions of a designated set of VR modules on the da Vinci Skills Simulator to identify optimal VR warm-up curricula to prime technical skill. After performing their curricula, warm-up effect was assessed based on performance on a criterion task. The optimal warm-up curriculum was chosen from the group with the best task time and video review-based technical skill. Robot-assisted surgery-experienced surgeons were then recruited to either receive or not receive warm-up before surgery. Skill in the first 15 min of surgery was assessed by blinded surgeon and crowdworker review as well as tool motion metrics. The intervention was performing VR warm-up before human robot-assisted surgery. Warm-up effect was measured using objective performance metrics and video review using the Global Evaluative Assessment of Robotic Skills tool. Linear mixed effects models with a random intercept for each surgeon and nonparametric modified Friedman tests were used for analysis. RESULTS: The group performing only a Running Suture task on the simulator was on average 31.3 s faster than groups performing other simulation tasks and had the highest Global Evaluative Assessment of Robotic Skills scores from 41 surgeons who participated. This was chosen as the optimal curriculum. Thereafter, 34 surgeons completed 347 surgeries with corresponding video and tool motion data. No statistically significant differences in skill were observed with the warm-up intervention. CONCLUSIONS: We conclude that a robotic VR warm-up before performing the early stages of surgery does not impact the technical skill of the surgeon.

The Effects of Hyperbaric Oxygen on Rheumatoid Arthritis: A Pilot Study.

BACKGROUND/OBJECTIVE: This case series pilot study assessed the effects of hyperbaric oxygen therapy (HBO2) for treating rheumatoid arthritis (RA). METHODS: Ten RA subjects received 30 HBO2 treatments over 6 to 10 weeks. Serial rheumatologic evaluations (ie, the Disease Activity Scale [DAS28], the Routine Assessment of Patient Index Data 3, and the Pain and Sleep Quality Questionnaire) were completed at baseline, throughout the course of the study, and at the 6-month follow-up. RESULTS: There was a statistically significant effect of HBO2 therapy over time on the DAS28-Global Health (p = 0.01), the DAS28-C-reactive protein (p = 0.002), and the DAS28-erythrocyte sedimentation rate (p = 0.008) measures; these analyses excluded 2 patients who were in clinical remission at baseline. Selected post hoc comparisons showed significantly lower DAS28-Global Health, DAS28-C-reactive protein, and DAS28-erythrocyte sedimentation rate scores at 3 and 6 months relative to baseline. In addition, statistically significant decreases in pain as measured by the Routine Assessment of Patient Index Data 3 and Pain and Sleep Quality Questionnaire were observed at the end of HBO2 relative to baseline. CONCLUSIONS: Hyperbaric oxygen therapy is effective for joint pain in patients with RA based on data from multiple, validated clinical measures. Further research with more subjects and the use of a control group is necessary.

Foxp3+ Tregs from Langerhans cell histiocytosis lesions co-express CD56 and have a definitively regulatory capacity.

Langerhans cell histiocytosis (LCH) lesions contain myeloid lineage 'LCH' cells. Regulatory T cells (Tregs) are also enriched within lesions, although their role in LCH pathogenesis is unknown. LCH cells are thought to produce the transforming growth factor beta (TGF-ß) within lesions, however whether Tregs contribute is unestablished. Using flow cytometry, we analyzed relative frequencies of live Tregs from LCH patients and identified CD56 expression and TGF-ß production by lesion Tregs. While CD56+ Tregs were enriched in lesions, overall CD56+ T cells were reduced in the blood from active LCH patients compared to non-active disease patients, and there was a negative correlation between CD8+CD56+ T cells and Tregs. We propose that inducing a Treg phenotype in T cells such as CD56+ T cells may be a mechanism by which LCH cells divert inflammatory T cell responses. Thus, Tregs within LCH lesions are likely an important component in LCH pathogenesis.

Chronically stimulated human MAIT cells are unexpectedly potent IL-13 producers.

Mucosal-associated invariant T (MAIT) cells are unconventional T cells that recognize antigens derived from riboflavin biosynthesis. In addition to anti-microbial functions, human MAIT cells are associated with cancers, autoimmunity, allergies and inflammatory disorders, although their role is poorly understood. Activated MAIT cells are well known for their rapid release of Th1 and Th17 cytokines, but we have discovered that chronic stimulation can also lead to potent interleukin (IL)-13 expression. We used RNA-seq and qRT-PCR to demonstrate high expression of the IL-13 gene in chronically stimulated MAIT cells, and directly identify IL-13 using intracellular flow cytometry and multiplex bead analysis of MAIT cell cultures. This unexpected finding has important implications for IL-13-dependent diseases, such as colorectal cancer (CRC), that occur in mucosal areas where MAIT cells are abundant. We identify MAIT cells near CRC tumors and show that these areas and precancerous polyps express high levels of the IL-13 receptor, which promotes tumor progression and metastasis. Our data suggest that MAIT cells have a more complicated role in CRC than currently realized and that they represent a promising new target for immunotherapies where IL-13 can be a critical factor.

Efficacy of a bacterial fluorescence imaging device in an outpatient wound care clinic: a pilot study.

OBJECTIVE: Subsurface bacterial burden can be missed during standard wound examination protocols. The real-time bacterial fluorescence imaging device, MolecuLight i:X, visualises the presence of potentially harmful levels of bacteria through endogenous autofluorescence, without the need for contrast agents or contact with the patient. The intended use of the imaging device is to assist with the management of patients with wounds by enabling real-time visualisation of potentially harmful bacteria. The aim of this study was to establish the accuracy of the wound imaging device at detecting pathogenic bacteria in wounds. METHODS: A single-centre, prospective observational study was conducted in Cork University Hospital in an outpatient plastic surgery wound care clinic. Patients had their wounds photographed under white and autofluorescent light with the imaging device. Auto-fluorescent images were compared with the microbiological swab results. RESULTS: A total of 33 patients and 43 swabs were included, of which 95.3% (n=41) were positive for bacteria growth. Staphylococcus aureus was the most common bacterial species identified. The imaging device had a sensitivity of 100% and specificity of 78% at identifying pathological bacteria presence in wounds on fluorescent light imaging. The positive predictive value (PPV) was 95.4%. The negative predictive value (NPV) was 100%. It demonstrated a sensitivity and specificity of 100% at detecting the presence of Pseudomonas spp. CONCLUSION: The imaging device used could be a safe, effective, accurate and easy-to-use autofluorescent device to improve the assessment of wounds in the outpatient clinic setting. In conjunction with best clinical practice, the device can be used to guide clinicians use of antibiotics and specialised dressings.

Radiographic Outcomes of Hemiepiphyseal Stapling for Distal Radius Deformity Due to Multiple Hereditary Exostoses.

BACKGROUND: Multiple hereditary exostoses (MHE) affect the distal forearm in 30% to 60% of patients with this condition. Many surgical interventions have been described, but treatment is controversial. We present the radiographic results of low risk, minimally invasive hemiepiphyseal stapling of the distal radius to correct the joint angulation caused by MHE through growth modulation. METHODS: A retrospective review was performed on 18 forearms affected by MHE that underwent radial hemiepiphyseal stapling with follow-up of ≥2 years. The radial articular angle, carpal slip, ulnar tilt, lunate subsidence, and metaphyseal epiphyseal angle were measured on preoperative and final postoperative radiographs and compared. RESULTS: Statistically significant (P<0.05) improvements were found in 4 of 5 measurements, including the radial articular angle, carpal slip, ulnar tilt, and metaphyseal epiphyseal angle. No statistical difference was found in lunate subsidence (P=0.84). CONCLUSION: Hemiepiphyseal stapling of the radial side of the distal radius is a simple and effective method for correcting the radiographic deformity of the distal radius and may be an attractive option for the treatment of the distal radial deformity that is characteristic of the disease. LEVEL OF EVIDENCE: Level IV­therapeutic case series.

Cerebrovascular Accident in a High-Risk Patient During the Early Initiation Phase With Canagliflozin.

OBJECTIVE: To report a case of cerebrovascular accident (CVA) in a high-risk patient following initiation of canagliflozin, the first-in-class sodium-glucose-co-transporter 2 inhibitor approved by the Food and Drug Administration for type 2 diabetes mellitus. CASE SUMMARY: We describe a 62-year-old woman, with multiple clinical risk factors for stroke, who began canagliflozin 300 mg daily in addition to basal insulin therapy for diabetes management. The patient developed expressive aphasia 15 days following initiation of canagliflozin. Neuroimaging revealed acute infarcts of the left basal ganglia and temporal and parietal lobes. The patient was diagnosed with a CVA. Canagliflozin therapy was discontinued, metformin therapy was reinitiated in addition to the patient's basal insulin, and the patient was treated with antiplatelet, statin, and speech therapies. DISCUSSION: Assessment of the cardiovascular (CV) safety of canagliflozin is currently being investigated. A numerical increase in CV events, including nonfatal stroke, has been noted in preliminary data from ongoing analyses of canagliflozin in patients with preexisting CV risk factors. Although significant clinical risk factors were present in the patient described, a workup for routine causality came back negative. According to the Naranjo probability score, initiation of canagliflozin had a possible to probable association with the patient's CVA. CONCLUSIONS: This case suggests a potential association between the timing of canagliflozin initiation and development of stroke in patients with multiple clinical risk factors. We advise practitioners to use caution when initiating this new agent in patients at high risk for stroke while long-term CV safety surveillance is ongoing.

Unmet need for medical care and safety net accessibility among Birmingham's homeless.

Although homeless individuals often experience health problems requiring care, there are limitations to available research concerning the scale of their needs and the accessibility of safety net agencies to meet them. Traditional access-to-care surveys calculate unmet need among all persons queried (rather than persons needing care), making it difficult to calculate what percentage of persons requiring care actually obtain it. Additionally, no research has compared the relative accessibility of safety net programs to homeless persons in need. This cross-sectional, community-based survey assessed the prevalence of unmet need for several specific types of health care and compared the accessibility of agencies in Birmingham, AL. Substantial proportions of respondents reported unmet needs for general medical care (46 %), specialty care (51 %), mental health care (51 %), dental care (62 %), medications (57 %), and care of a child (23 %). The most commonly mentioned sites where care was sought included a federally funded Health Care for the Homeless (HCH) program (59 %), a religious free clinic (31 %), and a public hospital emergency department (51 %). The HCH program was most commonly cited as the location where care, once sought, could not be obtained (15 %), followed by the county hospital primary care clinics (13 %). In this survey, unmet need was common for all types of care queried, including primary care. Key components of the safety net, including a federally funded homeless health care program, had suboptimum accessibility.

Intraoperative Radiation Therapy: A Large Integrated Health Care System's Approach and Outcomes.

Introduction Intraoperative radiation therapy (IORT) may not be as effective in the community compared with clinical trials. Methods The authors reviewed data from the electronic health records of patients who received IORT between February 2014 and February 2020 at a single center within a large integrated health care system. The primary outcome was ipsilateral breast tumor recurrence. Results Of 5731 potentially eligible patients, 245 (4.3%) underwent IORT (mean age: 65.4 ± 0.4 years; median follow-up time: 3.5 years ± 2.2 months). According to the American Society for Radiation Oncology's accelerated partial breast irradiation guidelines based on final pathology, 51% of patients were suitable candidates for IORT, 38.4% were cautionary, and 10.6% were unsuitable. For adjuvant therapy, 6.5% had consolidative whole breast irradiation, and 66.4% received endocrine treatment. At the median follow-up time of 3.5 years, overall ipsilateral breast tumor recurrence was 3.7%. Recurrences tended to be more frequent in patients who refused or did not complete endocrine treatment than in those who received it (7.4% vs 1.9%, p = 0.07). The complication rate was 14.7%, with seroma being the most common (8.2%). Discussion The IORT ipsilateral breast tumor recurrence rate of 3.7% confirms a higher-than-expected rate compared to randomized clinical trials, possibly due to less compliance with endocrine therapy. Conclusion The authors subsequently revised their IORT protocol to require endocrine treatment as a part of the IORT treatment plan and to strongly recommend adjuvant whole breast irradiation for all patients deemed cautionary or unsuitable for IORT according to the American Society for Radiation Oncology's accelerated partial breast irradiation guidelines.

Validation of Ideal Breast Characteristics With Breast Augmentation Patients.

Background: Breast augmentation procedures are one of the most commonly performed aesthetic procedures in the United States. Little work has focused on the general public's overall perception of the ideal breast or has validated them with patient photographs. Objectives: To validate crowdsourced perceptions of breasts with their alignment to the aesthetics of breast augmentation patients. Methods: A prospective cross-sectional study was performed using participants enrolled through the AmazonMechanical Turk crowdsourcing platform (Amazon Web Services, Amazon, Seattle, WA) to obtain participant opinions ofhow closely patient breasts aligned with previously obtained results of 4 ideal breast characteristics. Outcomes were reported based on the correlation between breast attractiveness and alignment to ideal breast characteristics, both before and after breast implant procedures. Results: 2306 responses from 737 participants reported patient photograph alignment with ideal breast projection proportion (1.0) as having the highest correlation to opinions of heightened aesthetic beauty (R = 0.98, P < 0.001), and ideal nipple direction (front) as having the lowest correlation to aesthetic beauty (R = 0.90, P < 0.001). Younger age groups (18-24) and participants with a high school diploma or less rated patients as less attractive, while married and wealthy individuals reported higher attraction levels. Conclusions: Crowdsourcing can be a useful tool for aesthetic surgery preferences and has helped reveal key takeaways. The importance of the 4 breast characteristics has been validated, with alignment to all 4 characteristics tested having a high correlation to preferences. Differences in preference across demographic groups are a topic to further investigate.

Public Perception of Ideal Breast Shape.

BACKGROUND: In the past decade, there has been a dramatic increase in trends related to body-shaping procedures. According to the American Society of Plastic Surgeons, nearly 300,000 breast augmentation procedures were conducted in 2019. Learning the ideal shape of a breast and which esthetics lead to public perception of the most attractive breast is beneficial to properly performing these procedures. OBJECTIVES: The authors aimed to quantify the public's perception of attraction to breast shape by measuring public opinion for various esthetic elements of breast anatomy and linking this to various demographic factors. METHODS: Survey responses were collected from 1000 users of Amazon Mechanical Turk to collect demographic data and ask users to rank preferences for randomized image panels of breast proportions. RESULTS: 960 responses were used for analysis. A majority of respondents were male (60%), with a plurality being 25 to 34 years old (49.3%). The most notable preferences between all groups were breast projection proportion and nipple direction, with preferences of 1.0 and frontal nipple direction, respectively. Breast width to shoulder width ratio also had a clear preference among the crowd, with 105% being the preferred percentage, and the 25 to 34 age group having a very strong preference for this. CONCLUSIONS: The authors used a crowdsourcing survey technique with randomized image panels to analyze ideal breast preference using images of various anatomical traits of the female breast. It was concluded that crowdsourcing can be a favorable technique for learning ideal overall preferences for specific anatomy.

Crowd-Sourced Reliability of an Assessment of Lower Facial Aging Using a Validated Visual Scale.

Reliable and valid assessments of the visual endpoints of aesthetic surgery procedures are needed. Currently, most assessments are based on the opinion of patients and their plastic surgeons. The objective of this research was to analyze the reliability of crowdworkers assessing de-identified photographs using a validated scale that depicts lower facial aging. METHODS: Twenty photographs of the facial nasolabial region of various non-identifiable faces were obtained for which various degrees of facial aging were present. Independent crowds of 100 crowd workers were tasked with assessing the degree of aging using a photograph numeric scale. Independent groups of crowdworkers were surveyed at 4 different times (weekday daytime, weekday nighttime, weekend daytime, weekend nighttime), once a week for 2 weeks. RESULTS: Crowds assessing midface region photographs had an overall correlation of R = 0.979 (weekday daytime R = 0.991; weekday nighttime R = 0.985; weekend daytime R = 0.997; weekend nighttime R = 0.985). Bland-Altman test for test-retest agreement showed a normal distribution of assessments over the various times tested, with the differences in the majority of photographs being within 1 SD of the average difference in ratings. CONCLUSIONS: Crowd assessments of facial aging in de-identified photographs displayed very strong concordance with each other, regardless of time of day or week. This shows promise toward obtaining reliable assessments of pre and postoperative results for aesthetic surgery procedures. More work must be done to quantify the reliability of assessments for other pretreatment states or the corresponding results following treatment.

A Modified MTS Proliferation Assay for Suspended Cells to Avoid the Interference by Hydralazine and ß-Mercaptoethanol.

The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay is one of the most commonly used tests of cell proliferation. Hydralazine has been reported to interfere with the performance of the MTS assay when used on adherent cells. This study aimed to investigate whether hydralazine interferes with the performance of the MTS assay on suspended cells. THP-1 (a monocytic leukemia cell line) cells were cultured in the presence or absence of hydralazine (0, 10, 50, 100, and 500 µM) for 2 or 24 h. Cell numbers were analyzed using the MTS, trypan blue exclusion, or microscopic assays. A modified version of the standard MTS assay was established by centrifuging the cells and replacing the test medium with fresh culture medium immediately before the addition of the MTS reagent. Culture of THP-1 cells with hydralazine at concentrations of 50, 100, and 500 µM for 2 h increased absorbance (p < 0.001) in the standard MTS assay, whereas both the trypan blue exclusion assay and microscopy suggested no change in cell numbers. Culture of THP-1 cells with 100 and 500 µm hydralazine for 24 h increased absorbance (p < 0.05) in the standard MTS assay; however, trypan blue exclusion and microscopy suggested a decrease in cell numbers. In a cell-free system, hydralazine (100 and 500 µM) increased absorbance in a time- and concentration-dependent manner. The modified MTS assay produced results consistent with trypan blue exclusion and microscopy using THP-1 cells. In addition, the modified MTS assay produced reliable results when K562 and Jurkat cells were incubated with hydralazine or ß-mercaptoethanol (ßME). In conclusion, a simple modification of the standard MTS assay overcame the interference of hydralazine and ßME when assessing suspended cells.

A Role for MAIT Cells in Colorectal Cancer.

MAIT cells are MR1-restricted T cells that are well-known for their anti-microbial properties, but they have recently been associated with different forms of cancer. Several studies have reported activated MAIT cells within the microenvironment of colorectal tumors, but there is conjecture about the nature of their response and whether they are contributing to anti-tumor immunity, or to the progression of the disease. We have reviewed the current state of knowledge about the role of MAIT cells in colorectal cancer, including their likely influence when activated and potential sources of stimulation in the tumor microenvironment. The prospects for MAIT cells being used in clinical settings as biomarkers or as targets of new immunotherapies designed to harness their function are discussed.

Temporal variability of surgical technical skill perception in real robotic surgery.

PURPOSE: Summary score metrics, either from crowds of non-experts, faculty surgeons or from automated performance metrics, have been trusted as the prevailing method of reporting surgeon technical skill. The aim of this paper is to learn whether there exist significant fluctuations in the technical skill assessments of a surgeon throughout long durations of surgical footage. METHODS: A set of 12 videos of robotic surgery cases from common human patient robotic surgeries were used to evaluate the perceived technical skill at each individual minute of the surgical videos, which were originally 12-15 min in length. A linear mixed-effects model for each video was used to compare the ratings of each minute to those from every other minute in order to learn whether a change in scores over time can be detected and reliably measured apart from inter- and intrarater variation. RESULTS: Modeling the change over time of the global evaluative assessment of robotic skills scores significantly contributed to the prediction models for 11 of the 12 surgeons. This demonstrates that measurable changes in technical skill occur over time during robotic surgery. CONCLUSION: The findings from this research raise questions about the optimal duration of footage needed to be evaluated to arrive at an accurate rating of surgical technical skill for longer procedures. This may imply non-negligible label noise for supervised machine learning approaches. In the future, it may be necessary to report a surgeon's skill variability in addition to their mean score to have proper knowledge of a surgeon's overall skill level.