Laser ablation for spongiform thyroid nodules: identifying baseline nodule volume and energy to be delivered for successful treatment.

BACKGROUND: Laser ablation (LA) is a minimally invasive treatment. It has been widely used since the early 2000s to induce volume reduction of symptomatic benign thyroid nodules. Up to 40% of laser-treated nodules have been reported to achieve a volume reduction of <50% (technique inefficacy) at 12 months and tend to regrow over time. OBJECTIVE: This study aimed to assess the optimal baseline volume and energy to be delivered to minimize technique inefficacy. METHODS: This was a retrospective study. Data were collected, including baseline volume, energy delivered, and 12-month volume reduction ratio (VRR) of spongiform nodules (EU-TIRADS 2) treated with LA between 2010 and 2020. Based on these data, the optimal baseline volume and energy to be delivered were calculated to maximize the rate of nodules with technique efficacy (VRR ≥ 50% at 12-month follow-up). RESULTS: A total of 205 patients with spongiform nodules were included in this study. The energy delivered was positively associated with VRR. However, no association was observed between baseline volume and VRR. Delivering energy ≥500 J/mL to nodules with a mean baseline volume of 11.4 ± 4 mL resulted in technique efficacy in 83% of cases. CONCLUSION: Treating spongiform nodules with a baseline volume of ≤15 mL and delivering energy ≥500 J/mL are key factors for achieving a relevant rate of technique efficacy.

Rates of diagnostic and therapeutic procedures in a cohort of patients undergoing first endocrine consultation for thyroid nodular disease.

PURPOSE: This study aimed to evaluate the rates of necessary diagnostic and therapeutic procedures in a cohort of patients undergoing their first endocrine consultation for thyroid nodular disease. METHODS: This was an observational study conducted between January 1 and June 30, 2023, on patients undergoing their first endocrine consultation for thyroid nodular disease. Data were collected, including age, thyroid-stimulating hormone (TSH) concentration, reasons for performing thyroid ultrasound (US), and thyroid US reports. The US was performed at the time of the endocrine consultation according to the American College of Radiology Thyroid Imaging, Reporting and Data System (ACR-TIRADS) risk stratification system. Patients underwent further investigations/treatment or simple US follow-up as necessary. RESULTS: A total of 373 patients with thyroid nodules were evaluated. Reasons for undergoing thyroid US were unrelated to suspected thyroid disease in 126 (33.8%), incidentalomas in 91 (24.4%), dysfunction or positivity for thyroid antibodies in 67 (18%), symptoms or visible nodules in 61 (16.3%), and family history of thyroid disease in 28 (7.5%). A total of 193 diagnostic or therapeutic procedures were performed in 133 (35.7%) patients [fine-needle aspiration (FNA): 121 (62.7%), surgery: 28 (14.5%), percutaneous ethanol injection: 20 (10.4%), scintigraphy: 10 (5.2%); thermal ablation: 7 (3.6%), and radioactive treatment: 7 (3.6%)]. CONCLUSIONS: In the present study only one-third of the patients undergoing endocrine consultation with first detected thyroid nodules required any diagnostic or therapeutic procedures.

Patients undergoing endocrine consultation and first diagnosis of nodular disease: Indications of thyroid ultrasound and completeness of ultrasound reports.

PURPOSE: To evaluate reasons for performing ultrasonography (US) and completeness of US reports in patients undergoing endocrine consultation with the first diagnosis of nodular disease. METHODS: Since January 1 to June 30, 2021, we prospectively collected patient data (age and thyroid-stimulating hormone concentrations), reasons for performing thyroid US, and completeness of reports regarding the description of the thyroid gland and nodules. In the case of multiple nodules, we considered the nodule suspected of malignancy and the largest one. To evaluate the accuracy of thyroid nodule description, we referred to the five characteristics suggested by the ACR TI-RADS system. RESULTS: A total of 341 patients with thyroid nodules received endocrine consultation (female, 78%). The most frequent reasons for performing thyroid US were unrelated to a suspected thyroid disease (31.7%), followed by incidentaloma (23.5%), dysfunction or positivity for thyroid antibodies (19.1%), symptomatic or visible nodules (17.6%), and family history of any thyroid disease (8.2%). Gland texture was not reported in 41.9%. The depth of the lobes was the dimension reported most frequently (42.2%), but any diameter was not reported in 57.8% of the cases. As regards the description of the most relevant nodule, length was reported more frequently (75.9%). Margins and echogenicity were more frequently described (54.5% and 44.3%, respectively) than other characteristics (composition: 27%; shape: 8.8%; echogenic foci: 6.7%). No reports had indicated the malignancy risk stratification. CONCLUSIONS: The results of the study demonstrate that in patients undergoing endocrine consultation with first detected thyroid nodules, US was mostly performed in asymptomatic cases, US reports were incomplete, and no risk stratification system was reported.

Influence of experimental methods on the mechanical properties of silk fibers: A systematic literature review and future road map.

Spider silk fibers are of scientific and industrial interest because of their extraordinary mechanical properties. These properties are normally determined by tensile tests, but the values obtained are dependent on the morphology of the fibers, the test conditions, and the methods by which stress and strain are calculated. Because of this, results from many studies are not directly comparable, which has led to widespread misconceptions in the field. Here, we critically review most of the reports from the past 50 years on spider silk mechanical performance and use artificial spider silk and native silks as models to highlight the effect that different experimental setups have on the fibers' mechanical properties. The results clearly illustrate the importance of carefully evaluating the tensile test methods when comparing the results from different studies. Finally, we suggest a protocol for how to perform tensile tests on silk and biobased fibers.

Engineered Spider Silk Proteins for Biomimetic Spinning of Fibers with Toughness Equal to Dragline Silks.

Spider silk is the toughest fiber found in nature, and bulk production of artificial spider silk that matches its mechanical properties remains elusive. Development of miniature spider silk proteins (mini-spidroins) has made large-scale fiber production economically feasible, but the fibers' mechanical properties are inferior to native silk. The spider silk fiber's tensile strength is conferred by poly-alanine stretches that are zipped together by tight side chain packing in ß-sheet crystals. Spidroins are secreted so they must be void of long stretches of hydrophobic residues, since such segments get inserted into the endoplasmic reticulum membrane. At the same time, hydrophobic residues have high ß-strand propensity and can mediate tight inter-ß-sheet interactions, features that are attractive for generation of strong artificial silks. Protein production in prokaryotes can circumvent biological laws that spiders, being eukaryotic organisms, must obey, and the authors thus design mini-spidroins that are predicted to more avidly form stronger ß-sheets than the wildtype protein. Biomimetic spinning of the engineered mini-spidroins indeed results in fibers with increased tensile strength and two fiber types display toughness equal to native dragline silks. Bioreactor expression and purification result in a protein yield of ≈9 g L-1 which is in line with requirements for economically feasible bulk scale production.

Prey localization in spider orb webs using modal vibration analysis.

Spider webs are finely tuned multifunctional structures, widely studied for their prey capture functionalities such as impact strength and stickiness. However, they are also sophisticated sensing tools that enable the spider to precisely determine the location of impact and capture the prey before it escapes. In this paper, we suggest a new mechanism for this detection process, based on potential modal analysis capabilities of the spider, using its legs as distinct distributed point sensors. To do this, we consider a numerical model of the web structure, including asymmetry in the design, prestress, and geometrical nonlinearity effects. We show how vibration signals deriving from impacts can be decomposed into web eigenmode components, through which the spider can efficiently trace the source location. Based on this numerical analysis, we discuss the role of the web structure, asymmetry, and prestress in the imaging mechanism, confirming the role of the latter in tuning the web response to achieve an efficient prey detection instrument. The results can be relevant for efficient distributed impact sensing applications.

1000 spider silkomes: Linking sequences to silk physical properties.

Spider silks are among the toughest known materials and thus provide models for renewable, biodegradable, and sustainable biopolymers. However, the entirety of their diversity still remains elusive, and silks that exceed the performance limits of industrial fibers are constantly being found. We obtained transcriptome assemblies from 1098 species of spiders to comprehensively catalog silk gene sequences and measured the mechanical, thermal, structural, and hydration properties of the dragline silks of 446 species. The combination of these silk protein genotype-phenotype data revealed essential contributions of multicomponent structures with major ampullate spidroin 1 to 3 paralogs in high-performance dragline silks and numerous amino acid motifs contributing to each of the measured properties. We hope that our global sampling, comprehensive testing, integrated analysis, and open data will provide a solid starting point for future biomaterial designs.

Silk fibroin films with embedded magnetic nanoparticles: evaluation of the magneto-mechanical stimulation effect on osteogenic differentiation of stem cells.

We report about a biomaterial in the form of film ∼10 µm thick, consisting of a silk fibroin matrix with embedded iron oxide superparamagnetic nanoparticles, for prospective applications as bioactive coating in regenerative medicine. Films with different load of magnetic nanoparticles are produced (nanoparticles/silk fibroin nominal ratio = 5, 0.5 and 0 wt%) and the structural, mechanical and magnetic properties are studied. The nanoparticles form aggregates in the silk fibroin matrix and the film stiffness, as tested by nanoindentation, is spatially inhomogeneous, but the protein structure is not altered. In vitro biological tests are carried out on human bone marrow-derived mesenchymal stem cells cultured on the films up to 21 days, with and without an applied static uniform magnetic field. The sample with the highest nanoparticles/silk fibroin ratio shows the best performance in terms of cell proliferation and adhesion. Moreover, it promotes a faster and better osteogenic differentiation, particularly under magnetic field, as indicated by the gene expression level of typical osteogenic markers. These findings are explained in light of the results of the physical characterization, combined with numerical calculations. It is established that the applied magnetic field triggers a virtuous magneto-mechanical mechanism in which dipolar magnetic forces between the nanoparticle aggregates give rise to a spatial distribution of mechanical stresses in the silk fibroin matrix. The film with the largest nanoparticle load, under cell culture conditions (i.e. in aqueous environment), undergoes matrix deformations large enough to be sensed by the seeded cells as mechanical stimuli favoring the osteogenic differentiation.

The mechanical characterization of the legs, fangs, and prosoma in the spider Harpactira curvipes (Pocock 1897).

The exoskeleton of spiders is the primary structure that interacts with the external mechanical stimuli, thus playing a crucial role in spider life. In particular, fangs, legs, and prosoma are the main rigid structures of the exoskeleton and their properties must be measured to better understand their mechanical behaviours. Here we investigate, by means of nanoindentation, the mechanical properties of the external sclerotized cuticles of such parts in the spider Harpactira curvipes. Interestingly, the results show that the leg's cuticle is stiffer than the prosoma and has a stiffness similar to the one of the tip fangs. This could be explained by the legs' function in perceiving vibrations that could be facilitated by higher stiffness. From a broader perspective, this characterization could help to understand how the same basic material (the cuticle, i.e. mainly composed of chitin) can be tuned to achieve different mechanical functions, which improves the animal's adaptation to specific evolutive requirements. We, thus, hope that this work stimulates further comparative analysis. Moreover, these results may also be potentially important to inspire the design of graded materials with superior mechanical properties.

Artificial and natural silk materials have high mechanical property variability regardless of sample size.

Silk fibres attract great interest in materials science for their biological and mechanical properties. Hitherto, the mechanical properties of the silk fibres have been explored mainly by tensile tests, which provide information on their strength, Young's modulus, strain at break and toughness modulus. Several hypotheses have been based on these data, but the intrinsic and often overlooked variability of natural and artificial silk fibres makes it challenging to identify trends and correlations. In this work, we determined the mechanical properties of Bombyx mori cocoon and degummed silk, native spider silk, and artificial spider silk, and compared them with classical commercial carbon fibres using large sample sizes (from 10 to 100 fibres, in total 200 specimens per fibre type). The results confirm a substantial variability of the mechanical properties of silk fibres compared to commercial carbon fibres, as the relative standard deviation for strength and strain at break is 10-50%. Moreover, the variability does not decrease significantly when the number of tested fibres is increased, which was surprising considering the low variability frequently reported for silk fibres in the literature. Based on this, we prove that tensile testing of 10 fibres per type is representative of a silk fibre population. Finally, we show that the ideal shape of the stress-strain curve for spider silk, characterized by a pronounced exponential stiffening regime, occurs in only 25% of all tested spider silk fibres.

An Image-Analysis-Based Method for the Prediction of Recombinant Protein Fiber Tensile Strength.

Silk fibers derived from the cocoon of silk moths and the wide range of silks produced by spiders exhibit an array of features, such as extraordinary tensile strength, elasticity, and adhesive properties. The functional features and mechanical properties can be derived from the structural composition and organization of the silk fibers. Artificial recombinant protein fibers based on engineered spider silk proteins have been successfully made previously and represent a promising way towards the large-scale production of fibers with predesigned features. However, for the production and use of protein fibers, there is a need for reliable objective quality control procedures that could be automated and that do not destroy the fibers in the process. Furthermore, there is still a lack of understanding the specifics of how the structural composition and organization relate to the ultimate function of silk-like fibers. In this study, we develop a new method for the categorization of protein fibers that enabled a highly accurate prediction of fiber tensile strength. Based on the use of a common light microscope equipped with polarizers together with image analysis for the precise determination of fiber morphology and optical properties, this represents an easy-to-use, objective non-destructive quality control process for protein fiber manufacturing and provides further insights into the link between the supramolecular organization and mechanical functionality of protein fibers.

Image-guided thermal ablation in autonomously functioning thyroid nodules. A retrospective multicenter three-year follow-up study from the Italian Minimally Invasive Treatment of the Thyroid (MITT) Group.

OBJECTIVES: To report the results of a multicenter retrospective evaluation of the clinical outcomes of thermal ablation (TA) in a large series of autonomously functioning thyroid nodules (AFTN) with a follow-up protracted up to 3 years. METHODS: Patients treated with single TA for an AFTN in Italy were included. Changes in nodule volume, TSH values, and ongoing anti-thyroid therapy were assessed at the 2-, 6-, 12-, 24-, and 36-month follow-up controls. Complications and need of any additional therapy after TA were also registered. RESULTS: A total of 361 patients (244 females, 117 males, median age 58 years, IQR 46-70 years) were included. Nodule volume was significantly reduced at all time points (p < 0.001) (median volume reduction 58% at 6-month and 60% at 12-month). Serum TSH values increased significantly at all time points. After TA, anti-thyroid therapy was withdrawn in 32.5% of patients at 2 months, in 38.9% at 6 months, and in 41.3% at 12 months. A significant difference in the rate of patients who withdrawn medical therapy at 12 months was registered between small (< 10 mL) (74%), medium (49%), or large (> 30 mL) nodules (19%). A single major complication occurred (0.25%). Additional treatments were needed in 34/361 (9.4%) of cases including 4 (1.1%) surgical treatment. CONCLUSIONS: Image-guided thermal ablation offers a further safe and effective therapeutic option in patients with AFTN. Clinical outcomes are significantly more favorable in small than in large size AFTN. KEY POINTS: • Thermal ablations (TA) can be safely and effectively used in patients with autonomously functioning thyroid nodules (AFTN). • TA results in a clinically significant nodule volume reduction that is paralleled by TSH level normalization and anti-thyroid drug therapy discontinuation (after TA anti-thyroid therapy was withdrawn in 41.3% at 12 months). • Clinical outcomes after TA are more favorable in small nodules, and when a large amount of thyroid nodule tissue is ablated.

Tidy dataset of the experimental design of the optimization of the alkali degumming process of Bombyx mori silk.

Silk fibroin is the structural fiber of the silk filament and it is usually separated from the external protein, named sericine, by a chemical process called degumming. This process consists of an alkali bath in which the silk cocoons are boiled for a determined time. It is also known that the degumming process impacts the property of the outcoming silk fibroin fibers. In this work, we described the dataset obtained from a Design of Experiment (DoE) screening made on the alkali degumming. Four process factors were considered: the number of degumming baths, the process time, the process temperature, and the salt concentration. The data on the properties of the silk fibroin fibers were collected. In particular, the molecular weight was obtained by gel permeation chromatography (GPC), the mechanical data by tensile test and the secondary structure by Fourier Infrared Transform Spectroscopy (FTIR).

How spiders hunt heavy prey: the tangle web as a pulley and spider's lifting mechanics observed and quantified in the laboratory.

The spiders of Theridiidae's family display a peculiar behaviour when they hunt extremely large prey. They lift the quarry, making it unable to escape, by attaching pre-tensioned silk threads to it. In this work, we analysed for the first time in the laboratory the lifting hunting mechanism and, in order to quantify the phenomenon, we applied the lifting mechanics theory. The comparison between the experiments and the theory suggests that, during the process, spiders do not stretch the silk too much by keeping it in the linear elastic regime. We thus report here further evidence for the strong role of silk in spiders' evolution, especially how spiders can stretch and use it as an external tool to overcome their muscles' limits and capture prey with large mass, e.g. 50 times the spider's mass.

A Design of Experiment Rational Optimization of the Degumming Process and Its Impact on the Silk Fibroin Properties.

Silk fibroin is a protein with a unique combination of properties and is widely studied for biomedical applications. The extraction of fibroin (degumming) from the silk filament impacts the properties of the outcoming material. The degumming can be conducted with different procedures. Among them, the most used and studied procedure in the research field is the alkali degumming with sodium carbonate (Na2CO3). In this study, by the use of a statistical method, namely, design of experiment (DOE), we characterized the Na2CO3 degumming, taking into consideration the main process factors involved and changing them within a selected range of values. We considered the process temperature and time, the salt concentration, and the number of baths used, testing the impact of these variables on the fibroin properties by building empirical models. These models not only took into consideration the direct effect of the process factors but also their combined effect, which are not conventionally detectable with other methods. The weight loss and the amount of sericin removed in the process were determined and used as a measure of the effectiveness of the process. The secondary structure, the molecular weight, the diameter of fibers, and their morphology and mechanical properties were studied with the intent to correlate the macroscopical properties with the structural changes. We report, for the first time, the possibility to effectively remove all sericin from the silk fibroin using Na2CO3, using a process that requires less salt, water, and energy, in comparison with the standard alkali protocol, making this technique overall more environmentally sustainable; in addition, we have demonstrated the possibility to tune the material properties by varying the degumming conditions and even to optimize them with empirical statistically based equations that allow one to directly set the optimal process parameters. The major effect on the macroscopical properties (such as the ultimate strength and Young's modulus) has been proved to be correlated with the removal of sericin instead of the microstructural variations. Finally, a ready-to-use table with a set of optimized degumming procedures to maximize or minimize the studied properties was provided.

Properties of Biomimetic Artificial Spider Silk Fibers Tuned by PostSpin Bath Incubation.

Efficient production of artificial spider silk fibers with properties that match its natural counterpart has still not been achieved. Recently, a biomimetic process for spinning recombinant spider silk proteins (spidroins) was presented, in which important molecular mechanisms involved in native spider silk spinning were recapitulated. However, drawbacks of these fibers included inferior mechanical properties and problems with low resistance to aqueous environments. In this work, we show that ≥5 h incubation of the fibers, in a collection bath of 500 mM NaAc and 200 mM NaCl, at pH 5 results in fibers that do not dissolve in water or phosphate buffered saline, which implies that the fibers can be used for applications that involve wet/humid conditions. Furthermore, incubation in the collection bath improved the strain at break and was associated with increased ß-sheet content, but did not affect the fiber morphology. In summary, we present a simple way to improve artificial spider silk fiber strain at break and resistance to aqueous solvents.

Mechanical Properties and Weibull Scaling Laws of Unknown Spider Silks.

Spider silks present extraordinary mechanical properties, which have attracted the attention of material scientists in recent decades. In particular, the strength and the toughness of these protein-based materials outperform the ones of many man-made fibers. Unfortunately, despite the huge interest, there is an absence of statistical investigation on the mechanical properties of spider silks and their related size effects due to the length of the fibers. Moreover, several spider silks have never been mechanically tested. Accordingly, in this work, we measured the mechanical properties and computed the Weibull parameters for different spider silks, some of them unknown in the literature. We also measured the mechanical properties at different strain rates for the dragline of the species Cupiennius salei. For the same species, we measured the strength and Weibull parameters at different fiber lengths. In this way, we obtained the spider silk scaling laws directly and according to Weibull's prediction. Both length and strain rates affect the mechanical properties of spider silk, as rationalized by Weibull's statistics.

Twelve-Month Volume Reduction Ratio Predicts Regrowth and Time to Regrowth in Thyroid Nodules Submitted to Laser Ablation: A 5-Year Follow-Up Retrospective Study.

OBJECTIVE: Laser ablation is a therapeutic modality used to reduce the volume of large benign thyroid nodules. Unsatisfactory reduction and regrowth are observed in some treated nodules. The aim of the study was to evaluate the long-term outcomes of laser treatment for solid nodules during a 5-year follow-up period, the regrowth rate, and the predictive risk factors of nodule regrowth. MATERIALS AND METHODS: We retrospectively evaluated patients with benign, solid, cold thyroid nodules who underwent laser ablation and were followed-up for 5 years. According to the selection criteria, 104 patients were included (median baseline nodule volume, 12.5 mL [25.0-75.0%, 8-18 mL]; median energy delivered, 481.5 J/mL [25.0-75.0%, 370-620 J/mL]). Nodule volume, thyroid function test results, and ultrasound were evaluated at baseline and then annually after the procedure. RESULTS: Of 104 patients, 31 patients (29.8%) had a 12-month volume reduction ratio (VRR) < 50.0% and 39 (37.5%) experienced nodule regrowth. Of these 39 patients, 17 (43.6%) underwent surgery and 14 (35.9%) underwent a second laser treatment. The rate of nodule regrowth was inversely related to the 12-month VRR, i.e., the lower the 12-month VRR, the higher the risk of regrowth (p < 0.001). The mean time for nodule regrowth was 33.5 ± 16.6 months. The 12-month VRR was directly related to time to regrowth, i.e., the lower the 12-month VRR, the shorter the time to regrowth (p < 0.001; R² = 0.3516). Non-spongiform composition increased the risk of regrowth with an odds ratio of 4.3 (95% confidence interval [CI] 1.8-10.2; p < 0.001); 12-month VRR < 50.0% increased the risk of regrowth with an odds ratio of 11.7 (95% CI 4.2-32.2; p < 0.001). CONCLUSION: The VRR of thyroid nodules subjected to similar amounts of laser energy varies widely and depends on the nodule composition; non-spongiform nodules are reduced to a lesser extent and regrow more frequently than spongiform nodules. A 12-month VRR < 50.0% is a predictive risk factor for regrowth and correlates with the time to regrowth.

The role of hairs in the adhesion of octopus suckers: a hierarchical peeling approach.

Organisms like the octopus or the clingfish are a precious source of inspiration for the design of innovative adhesive systems based on suction cups, but a complete mechanical description of their attachment process is still lacking. In this paper, we exploit the recent discovery of the presence of hairs in the acetabulum roof of octopus suction cups to revise the current model for its adhesion to the acetabulum wall. We show how this additional feature, which can be considered an example of a hierarchical structure, can lead to an increase of adhesive strength, based on the analysis of the cases of a simple tape and an axisymmetrical membrane adhering to a substrate. Using peeling theory, we discuss in both cases the influence of hierarchical structure and the resulting variation of geometry on the adhesive energy, highlighting how an increase in number of hierarchical levels contributes to its increment, with a corresponding improvement in functionality for the octopus suckers.