Nevin Manimala

Eur Biophys J. 2021 Jul 25. doi: 10.1007/s00249-021-01563-z. Online ahead of print.

ABSTRACT

Single-cell adhesion measured with atomic force microscopy (AFM) offers outstanding time and force resolution and allows the investigation of many important phenomena with unmatched precision. However, this technique suffers from serious practical limitations that hinder its effective application to a broader set of situations. Here we propose a different strategy based on the fabrication of large cantilevers and on the culture of the cells directly on them. Cantilevers are fabricated by standard micromachining, with an active area of 300 × 300 µm. A wedged structure is created so that the cantilever surface lies parallel to the substrate when mounted on an AFM system, so that the adhesion measurement probes the whole surface area at the same time. Thanks to the large area, cells can be seeded and grown on the cantilevers the day before the experiment, and let recover to optimal condition for the experiment. We used Human Embryonic Kidney cells, HEK 293A, to demonstrate the measurement of adhesion forces of up to 100 cells in parallel, and obtain a straightforward measurement of the average single cell adhesion energy. Our approach can improve significantly the cell-cell and cell-substrate adhesion statistics, reduce the experiment time and allow the investigation of the adhesion properties of cells that do not grow well in solution or on low adherent substrates, or that develop their characteristic features only after several hours or days of culture on a solid and adherent substrate.

PMID:34304293 | DOI:10.1007/s00249-021-01563-z

Support Care Cancer. 2021 Jul 25. doi: 10.1007/s00520-021-06415-2. Online ahead of print.

ABSTRACT

PURPOSE: Endometrial cancer is strongly linked to obesity and inactivity; however, increased physical activity has important benefits even in the absence of weight loss. Resistance (strength) training can deliver these benefits; yet few women participate in resistance exercise. The purpose of this study was to describe both physiological and functional changes following a home-based strength training intervention.

METHODS: Forty post-treatment endometrial cancer survivors within 5 years of diagnosis were enrolled in a pilot randomized trial, comparing twice-weekly home-based strength exercise to wait list control. Participants conducted the exercises twice per week for 10 supervised weeks with 5 weeks of follow-up. Measures included DXA-measured lean mass, functional fitness assessments, blood biomarkers, and quality of life outcomes.

RESULTS: On average, participants were 60.9 years old (SD = 8.7) with BMI of 39.9 kg/m² (SD = 15.2). At baseline, participants had 51.2% (SD = 6.0) body fat, which was not different between groups. Improvements were seen in the 30-s chair sit to stand (d = .99), the 30-s arm curl (d = .91), and the 8-ft up-and-go test (d = .63). No changes were measured for HbA1c or C-reactive protein. No changes were observed for flexibility (chair sit and reach, back scratch tests), 6-min walk test, maximum handgrip test, anxiety, depression, fatigue, or self-efficacy for exercise.

CONCLUSIONS: Home-based muscle-strengthening exercise led to favorable and clinically relevant improvements in 3 of 7 physical function assessments. Physical function, body composition, blood biomarkers, and patient-reported outcomes were feasible to measure. These fitness improvements were observed over a relatively short time frame of 10 weeks.

PMID:34304292 | DOI:10.1007/s00520-021-06415-2

Abdom Radiol (NY). 2021 Jul 25. doi: 10.1007/s00261-021-03225-2. Online ahead of print.

ABSTRACT

PURPOSE: To assess chronic liver disease (CLD) using multiparametric US in a private practice setting in a cohort of patients with increased skin-to-liver distance.

METHODS: 110 consecutive patients with increased skin-to-liver distance scheduled for US assessment of CLD were reviewed for study completion time, liver stiffness values (LS), attenuation imaging, and shear wave dispersion slope. The ROI was placed 2 cm below the liver capsule. The study included patients with NAFLD/NASH (68), hepatitis C (30), prior Fontan surgery (1), elevated liver function tests (5), alcohol abuse (3), hepatitis B (2), and primary biliary cirrhosis (1). IQR/M values were obtained. Comparison of less experienced sonographers (LES) and more experienced sonographers (MES) were evaluated through Student’s t test for independent data. Pearson coefficient r of correlation among quantitative variables was calculated.

RESULTS: The mean time to perform the exam was 129.7 ± 62.1 s. There was a statistically significant difference between LES and MES. The mean IQR/M for LS was 12.3 ± 5.5% m/s. Overall, in a cohort of difficult patients, 4.5% of LS values were not reliable. Fat quantification using attenuation imaging had a mean value of 0.60 ± 0.15 dB/cm/MHz (range 0.35-0.98 cm/dB/MHz) with an IQR/M of 14.7 ± 9.2%. Less reliable measurements of steatosis were obtained in 4.5% of patients. The mean shear wave dispersion slope was 12.74 ± 4.05 (m/s)/kHz (range 7.7-27.5 (m/s)/kHz) with an IQR/M of 38.7 ± 20.2% (range 3-131%). 20.9% of patients had values suggestive of compensated advanced chronic liver disease (cACLD).

CONCLUSION: Multiparametric US can provide assessment of CLD in less than 3 min in most patients and identify patients at risk for cACLD.

PMID:34304291 | DOI:10.1007/s00261-021-03225-2

Ann Hematol. 2021 Jul 24. doi: 10.1007/s00277-021-04558-0. Online ahead of print.

ABSTRACT

We conducted a phase II clinical trial to develop an autologous EBV-specific T cell product (baltaleucel T) for advanced, relapsed ENKTL. Among 47 patients who provided whole blood starting material for manufacturing the product, 15 patients received a median of 4 doses of baltaleucel T. Thirty-two (68%) patients did not receive baltaleucel-T due to manufacturing failure, rapid disease progression, and death. Of the 15 patients, 10 patients had measurable disease at baseline (salvage cohort), and 5 patients had no disease at baseline assessment (adjuvant cohort). In the 15 patients, the median follow-up duration was 10.2 months (range 2.0-23.5 months), median progression-free survival (PFS) was 3.9 months, and the median overall survival (OS) was not reached. Patients in the salvage cohort achieved a 30% complete response (CR) and a 50% overall response rate (ORR). In the adjuvant cohort, disease progression was reported in three patients and two patients did not relapse during study follow-up. When we compared survival outcomes of seven responders and eight non-responders, the PFS (P = 0.001) and OS (P = 0.014) of responders proved statistically superior to that of non-responders. Baltaleucel-T was well tolerated. We have performed a phase II clinical trial of autologous EBV-specific T cell treatment (baltaleucel-T) in R/R ENKTL. Autologous EBV-specific T cells were well tolerated and demonstrated single-agent activity in R/R ENTKL.

PMID:34304287 | DOI:10.1007/s00277-021-04558-0

Anal Bioanal Chem. 2021 Jul 25. doi: 10.1007/s00216-021-03536-9. Online ahead of print.

ABSTRACT

Immuno-mass spectrometry imaging (iMSI) uses laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) to determine the spatial expression of biomolecules in tissue sections following immunolabelling with antibodies conjugated to a metal reporter. As with all immunolabelling techniques, the binding efficiency of multiplexed staining can be affected by a number of factors including epitope blocking and other forms of steric hindrance. To date, the effects on the binding of metal-conjugated antibodies to their epitopes in a multiplexed analysis have yet to be quantitatively explored by iMSI. Here we describe a protocol to investigate the effects of multiplexing on reproducible binding using the muscle proteins, dystrophin, sarcospan, and myosin as a model, with antibodies conjugated with Maxpar® reagents before histological application to murine quadriceps sections using standard immunolabelling protocols and imaging with LA-ICP-MS. The antibodies were each individually applied to eight sections, and multiplexed to another eight sections. The average concentrations of the lanthanide analytes were determined, before statistical analyses found there was no significant difference between the individual and multiplexed application of the antibodies. These analyses provide a framework for ensuring reproducibility of antibody binding during multiplexed iMSI, which will allow quantitative exploration of protein-protein interactions and provide a greater understanding of fundamental biological processes during healthy and diseased states.

PMID:34304281 | DOI:10.1007/s00216-021-03536-9

Rom J Intern Med. 2021 Jul 23. doi: 10.2478/rjim-2021-0028. Online ahead of print.

ABSTRACT

INTRODUCTION: Thyroid hormones affect the cardiovascular system, but the precise mechanisms of their effects on the development of atherosclerosis are not entirely clear. The relationship between subclinical hypothyroidism, dyslipidemia and carotid atherosclerosis has been widely investigated, but the findings were controversial. The aim of the present study was to determine whether female subjects with subclinical hypothyroidism (SHypo) have increased carotid intima-media thickness (IMT) compared with euthyroid subjects, as well as to examine the association of SHypo, carotid atherosclerosis and dyslipidemia.

METHODS: This research included 100 women aged 30 to 70 years who were divided into two groups: the SHypo group including subjects with signs of subclinical hypothyroidism and the control group of euthyroid subjects. Carotid IMT thickness using B mode ultrasound was determined and its correlation with serum concentrations of fT4, TSH, CRP, and lipid profile including small dense LDL (sdLDL) was analysed.

RESULTS: Subjects with SHypo had significantly increased carotid IMT (IMT ≥ 75th centile) in all three measured segments of the right (p <0.001) and the left (p = 0.001) carotid artery compared to the control group, with a significantly more frequent appearance of plaque (25% vs. 9%; p = 0.05). Significant positive correlations were found between higher TSH and higher IMT values. Increased age, LDL cholesterol, and TSH predicted thickening of the carotid artery IMT.

CONCLUSIONS: It is important to screen people showing early, subclinical signs of thyroid gland dysfunction due to its impact on carotid atherosclerosis development.

PMID:34303321 | DOI:10.2478/rjim-2021-0028

Horm Mol Biol Clin Investig. 2021 Jul 26. doi: 10.1515/hmbci-2021-0012. Online ahead of print.

ABSTRACT

OBJECTIVES: As a systemic inflammatory disease, rheumatoid arthritis (RA) is the most common inflammatory arthritis in the population and there is no specific diagnostic marker in laboratory tests. The purpose of the study was to determine whether serum neopterin and pentraxin 3 (PTX3) levels may be a marker of increased inflammation in RA patients.

MATERIALS AND METHODS: The study were consist of 30 RA patients and 30 healthy controls who were admitted to the department of rheumatology. Blood specimens were taken from both group, and the levels of neopterin were analyzed by chromatography method (HPLC) and the PTX 3 levels were measured by enzyme-linked immunosorbent assay (ELISA). All data and demographic characteristics of participants were also recorded.

RESULTS: Serum neopterin and PTX 3 levels of the patient group (25.99 ± 7.24 ng/mL and 4.19 ± 1.01 ng/dL, respectively) was higher than the control group (9.55 ± 0.74 ng/mL and 2.23 ± 0.39 ng/dL, respectively). These results were remarkable significant (p<0.01). No statistically significant correlation was found between age-PTX 3, age-neopterin and PTX 3-neopterin parameters in the patient group. In the control group, a significant negative correlation was found between age and PTX 3 (p<0.05), and a positive correlation between neopterin and PTX 3.

CONCLUSIONS: Consequently, the serum neopterin and PTX 3 levels were higher in RA patients as compared to the healthy individuals. Our study suggest that there is a relation between neopterin and PTX 3 levels with RA patients. These findings suggest that neopterin and PTX 3 are important markers in the monitoring of RA disease.

PMID:34303320 | DOI:10.1515/hmbci-2021-0012

Eur J Dent. 2021 Jul 24. doi: 10.1055/s-0041-1729454. Online ahead of print.

ABSTRACT

OBJECTIVE: The study aimed to evaluate the cleaning efficacy of irrigant activation with a new ultrasonic tip in root-end preparations and to determine its influence on the bond strength of calcium silicate-based material.

MATERIALS AND METHODS: Maxillary canines were prepared and filled, and their root ends resected. Root-end cavities were ultrasonically prepared and randomly distributed into four groups according to the final irrigation protocols: G1 (ultrasonic irrigation [UI] + saline solution [SS]), G2 (syringe irrigation [SI] + SS), G3 (UI + ethylenediaminetetraacetic acid [EDTA]), and G4 (SI + EDTA). Cleaning efficacy analysis employed 72 specimens (n = 18) split longitudinally for imaging of the same areas by scanning electron microscopy (SEM). The percentage of dentinal tubules opened before and after irrigation was used as evaluation parameter. Push-out testing employed 40 specimens (n = 10) sectioned apical region perpendicularly, which slice was placed on a testing machine for the bond strength measurement and failure mode was assessed by SEM. The data were statistically analyzed (α ≤ 0.05).

RESULTS: G3 (UI + EDTA) removed the smear layer more effectively, showed the best tubule opening (p < 0.05), and presented the highest mean bond strength values (p < 0.05). Failure modes were predominantly adhesive, except for the G3 (UI + EDTA) group, in which they were mainly mixed (80%).

CONCLUSION: The results of this study suggest that EDTA 17% agitation promoted better cleaning and smear layer removal, improving the push-out bond strength of calcium silicate material in retrograde obturation.

PMID:34303319 | DOI:10.1055/s-0041-1729454

Eur J Dent. 2021 Jul 24. doi: 10.1055/s-0041-1726670. Online ahead of print.

ABSTRACT

OBJECTIVES: The aims of the study were to assess smile attractiveness of a collection of 68 smiling photographs of successfully treated cases submitted to the American Board of Orthodontics (ABO) clinical examination and identify variables that influence the assessment.

MATERIALS AND METHODS: A panel of 81 non-Caucasian assessors from various clinical disciplines were instructed to score the smile attractiveness on a visual analog scale from 1 (least attractive) to 10 (most attractive) and to select which components contributed to a lesser attractive smile. The mean, standard deviations (SDs), and quartiles of the smile attractiveness were obtained with descriptive statistics. Multilinear regression analysis was performed to investigate the scores of the perceived quality of smile attractiveness when the clinical disciplines and gender of the assessors were the factors taken into consideration. Receiver operating characteristic (ROC) curve was generated to establish the relationship between smile attractiveness and the achievement of a perfect smile.

RESULTS: The mean (SD) rating of each clinical photograph of the anterior occlusion on smiling ranged from 3.11 (1.47) as the least attractive smile to 7.59 (1.45) as the most attractive smile. The overall mean (SD) score for smile attractiveness was 5.30 (1.10). Problems associated with teeth, gingiva, and lips corresponded with a reduction of the smile attractiveness score by 1.56, 1.82, and 1.47, respectively. Gender was not associated with smile attractiveness ratings. Orthodontists, periodontists, and prosthodontists demonstrated no difference in the ratings, while plastic surgeons were more critical than orthodontists regarding smile attractiveness.

CONCLUSIONS: The study suggested that only 2 out of 68 AOB validated treatment finishes had a perfect and attractive smile.

PMID:34303318 | DOI:10.1055/s-0041-1726670

Eur J Dent. 2021 Jul 24. doi: 10.1055/s-0041-1729458. Online ahead of print.

ABSTRACT

OBJECTIVE: The study aimed to evaluate the influence of smoking habit on the prevalence of dental caries lesions in a follow-up study.

MATERIALS AND METHODS: A total of 3,675 patients (2,186 females and 1,489 males) with an average age of 51.4 years were included. Outcome measures were the incidence of dental caries defined as incipient noncavitated, microcavitated, or cavitated lesions which had been diagnosed through clinical observation with mouth mirror and probe examination evaluating change of texture, translucency, and color; radiographic examination through bitewing radiographs; or secondary caries through placement of a new restoration during the follow-up of the study.

STATISTICAL ANALYSIS: Cumulative survival (time elapsed with absence of dental caries) was estimated through the Kaplan-Meier product limit estimator with comparison of survival curves (log-rank test). A multivariable Cox proportional hazards regression model was used to evaluate the effect of smoking on the incidence of dental caries lesions when controlled to age, gender, systemic status, frequency of dental hygiene appointments, and socioeconomic status. The significance level was set at 5%.

RESULTS: Eight hundred sixty-three patients developed caries (23.5% incidence rate). The cumulative survival estimation was 81.8% and 48% survival rate for nonsmokers and smokers, respectively (p < 0.001), with an average of 13.5 months between the healthy and diseased state diagnosis. Smokers registered a hazard ratio for dental caries lesions of 1.32 (p = 0.001) when controlled for the other variables of interest.

CONCLUSION: Within the limitations of this study, it was concluded that smoking habit might be a predictor for dental caries.

PMID:34303317 | DOI:10.1055/s-0041-1729458