One-Pot Microfluidics to Engineer Chitosan Nanoparticles Conjugated with Antimicrobial Peptides Using "Photoclick" Chemistry: Validation Using the Gastric Bacterium Helicobacter pylori.

Surface bioconjugation of antimicrobial peptides (AMP) onto nanoparticles (AMP-NP) is a complex, multistep, and time-consuming task. Herein, a microfluidic system for the one-pot production of AMP-NP was developed. Norbornene-modified chitosan was used for NP production (NorChit-NP), and thiolated-AMP was grafted on their surface via thiol-norbornene "photoclick" chemistry over exposure of two parallel UV LEDs. The MSI-78A was the AMP selected due to its high activity against a high priority (level 2) antibiotic-resistant gastric pathogen: Helicobacter pylori (H. pylori). AMP-NP (113 ± 43 nm; zeta potential 14.3 ± 7 mV) were stable in gastric settings without a cross-linker (up to 5 days in pH 1.2) and bactericidal against two highly pathogenic H. pylori strains (1011 NP/mL with 96 µg/mL MSI-78A). Eradication was faster for H. pylori 26695 (30 min) than for H. pylori J99 (24 h), which was explained by the lower minimum bactericidal concentration of soluble MSI-78A for H. pylori 26695 (32 µg/mL) than for H. pylori J99 (128 µg/mL). AMP-NP was bactericidal by inducing H. pylori cell membrane alterations, intracellular reorganization, generation of extracellular vesicles, and leakage of cytoplasmic contents (transmission electron microscopy). Moreover, NP were not cytotoxic against two gastric cell lines (AGS and MKN74, ATCC) at bactericidal concentrations. Overall, the designed microfluidic setup is a greener, simpler, and faster approach than the conventional methods to obtain AMP-NP. This technology can be further explored for the bioconjugation of other thiolated-compounds.

Uncovering microplastics contamination in canned seafood.

There is limited research on the occurrence of microplastics (MPs) in canned seafood. All types of canned seafood investigated in the present study were contaminated. After sample digestion in 30 % hydrogen peroxide, a total of 40 MPs were recovered. Fibers were the most common type, blue was the dominant colour, and Fourier Transform Infrared Spectroscopy (FTIR) identified polyester as the most common polymer. Considering all samples, an average of 3.5 ± 5.2 MPs/can was obtained, with octopus in tomato sauce and tuna in olive oil presenting the highest contamination (5.2 ± 7.5 MPs/can and 5.2 ± 5.1 MPs/can, respectively). Also, significant differences between the number of MPs in the seafood tissues and immersion liquids were verified. The present study demonstrates MPs occurrence in canned seafood, a potential contamination pathway for humans. More research on the different stages of the canning processing is vital for understanding MPs contamination in cans.

Estimating Type 2 Diabetes Prevalence: A Model of Drug Consumption Data.

Observational, cross-sectional prevalence studies are costly and time-consuming. The development of indirect methods estimating prevalence used to obtain faster, less-expensive, and more robust results would be an advantage for several healthcare applications. This study aimed to use the drug dispensing data from community pharmacies to estimate the prevalence of Type 2 Diabetes mellitus (T2DM) in the Portuguese population. A cross-sectional study was conducted using a database of dispensed medicines with an indication for Diabetes mellitus in 2018 and 2021, stratified by geographic region. The methodology was based on a sequential method of acquiring prevalence estimates obtained through exposure to medicines using the daily doses defined per thousand inhabitants per day and adjusted to the rate of adherence to therapy, prescription patterns, and concomitance of antidiabetic drugs. The estimated overall T2DM prevalence in 2018 was 13.9%, and it was 14.2% for 2021. The results show the increased consumption of antidiabetic drugs, with fixed-dose combination antidiabetics and new antidiabetics being particularly important in 2021. This work allowed for the development of a model to obtain the estimated prevalence of T2DM based on drug consumption, using a simple, fast, and robust method that is in line with the available evidence. However, with the recent expanding indications for new antidiabetics, the inclusion of further data in the model needs to be studied.

Cie L*A*B* Values in VITA Classical and VITA 3D Master by Two Dental Spectrophotometers.

PURPOSE: To compare the CIE L*a*b* values of two different dental shade guides using two shade-matching spectrophotometers. MATERIALS AND METHODS: SpectroShade Micro (SS) and Easyshade (ES) (VITA Zahnfabrik) were tested. One calibrated operator performed 30 measurements of each tab from three batches of the two guides, VITA Classical (VC) and VITA 3D Shade Master (VM). Receiver operating characteristic (ROC) curves and intraclass coefficients (ICC) between the different batches were calculated. Results were given as mean and SD of the L*a*b* values and the respective color differences according to the CIEDE 2000 formula (ΔE00) for each of the shade tabs in the two spectrophotometers and then analyzed with independent student t test (α = .05). RESULTS: A total of 1,440 and 2,610 measurements for VC and VM, respectively, were performed per spectrophotometer. There were statistically significant differences between the two devices for all L*a*b* values, with the exception of: L* for 3R1.5; a* for 2R1.5, 3L1.5, 2L2.5, 3M2, 3L2.5, 4L1.5, and 4L2.5; and b* for D3, 1M2, and 3M2. When assessing the same shade guide, differences in ΔE00 were detected between devices above the acceptability threshold (AT) (ΔE00 ≥ 1.8) for all shade tabs except for VC (C1, C2, D3, A3.5, C3, and A4) and VM (4M1, 3R2.5, 4L2.5, and 4M3). The overall mean of the interdevice ΔE00 was 2.2 ± 1.0 for VC and 2.5 ± 1.0 for VM. CONCLUSIONS: The two dental spectrophotometers presented high ICC and ROC values, which validates their indication as auxiliary tools. However, there are discrepancies in the hues D (VC) and R (VM) with interdevice variability when evaluated for the L*a*b* component. Int J Prosthodont 2023;36:e38-e52.

Different Methods for Assessing Tooth Colour-In Vitro Study.

Colour assessment using digital methods can yield varying results, and it is important for clinicians to recognize the potential variability intra and inter-device. This study aimed to compare the L*a*b* values of VITA Classical (VC) and VITA Toothguide 3D-MASTER (VM) guides using two methods, SpectroShade (SS) and eLAB. Thirty-four measurements per tab were performed by a single operator across three batches of each guide. Intraclass correlation coefficients (ICC) between batches were calculated. Values <0.5, 0.5-0.75, 0.75-0.9, and >0.90 were classified as poor, moderate, good, and excellent reliability, respectively. Results were reported as mean and standard deviation of the L*a*b* values and respective colour differences (ΔE00) for each tab and method. Statistical analyses were performed with an independent t-test, α = 0.05. ICC values between batches were excellent for all L*a*b*, except for a* component in eLAB. There were statistically significant differences between methods in most L*a*b* values. The intra-device mean ΔE00 was 0.5 ± 0.6 for VC, 0.5 ± 0.8 for VM in SS, 1.1 ± 0.8 for VC, 1.1 ± 0.9 for VM in eLAB. The mean ΔE00 inter-device was 4.9 ± 1.7 for VC, 5.0 ± 1.7 for VM. Both methods demonstrated good internal consistency, with high ICC values and low intra-device colour differences, but exhibited high variability between methods, higher for a* the component.

Hydrogen Peroxide Diffusion through Dental Tissues-In Vitro Study.

Whitening products commonly utilize hydrogen peroxide (HP) as an active principle, which can penetrate dental tissues with potential side effects due to its low molecular weight. This study aimed to evaluate the HP diffusion of two in-office whitening products, namely 6% VivaStyle Paint On Plus (VS) and Opalescence Boost 40% (OP), in different tooth types. Additionally, the influence of the area of exposure, dental tissue thickness and pulp chamber volume was assessed. Each group consisted of eighteen intact anterior (A), premolar (PM) and molar (M) human teeth, and a positive pulpal pressure model was employed. The samples were analyzed using spectrophotometry, and results were expressed as the mean and 95% confidence interval. Statistical tests and linear regression models were appropriately applied at α = 5%. The total HP (µg) retrieved was as follows: VS-A, 1.333 [1.214, 1.452]; OP-A, 1.538 [1.457, 1.620]; VS-PM, 1.208 [1.123, 1.291]; OP-PM, 3.628 [3.401, 3.855]; VS-M, 2.560 [2.297, 2.823]; and OP-M, 4.197 [3.997, 4.396], with statistically significant differences in diffusion kinetics between whitening products for PM and M. Several HP concentrations attained a minimum cytotoxicity value of 2.22 µg/mL. The regression model shows that OP exposed the pulp chamber to 1.421 µg of HP more than that of VS. Different whitening products can cause cytotoxic HP concentrations in the pulp chamber, with a higher risk observed in molars.

Skin models of cutaneous toxicity, transdermal transport and wound repair.

Skin is widely used as a drug delivery route due to its easy access and the possibility of using relatively painless methods for the administration of bioactive molecules. However, the barrier properties of the skin, along with its multilayer structure, impose severe restrictions on drug transport and bioavailability. Thus, bioengineered models aimed at emulating the skin have been developed not only for optimizing the transdermal transport of different drugs and testing the safety and toxicity of substances but also for understanding the biological processes behind skin wounds. Even though in vivo research is often preferred to study biological processes involving the skin, in vitro and ex vivo strategies have been gaining increasing relevance in recent years. Indeed, there is a noticeably increasing adoption of in vitro and ex vivo methods by internationally accepted guidelines. Furthermore, microfluidic organ-on-a-chip devices are nowadays emerging as valuable tools for functional and behavioural skin emulation. Challenges in miniaturization, automation and reliability still need to be addressed in order to create skin models that can predict skin behaviour in a robust, high-throughput manner, while being compliant with regulatory issues, standards and guidelines. In this review, skin models for transdermal transport, wound repair and cutaneous toxicity will be discussed with a focus on high-throughput strategies. Novel microfluidic strategies driven by advancements in microfabrication technologies will also be revised as a way to improve the efficiency of existing models, both in terms of complexity and throughput.

Organizational culture and the individuals' discretionary behaviors at work: a cross-cultural analysis.

Stating "how things are done here," organizations are defining their culture. Organizational Culture (OC) is the set of values, norms, goals, and expectations shared by all members of an organization that aids in improving their commitment and performance. On the organizational level, it impacts behavior, productivity, and long-term survival by influencing organizational capability. Due to employee behavior being a competitive differential, this study examines how specific OCs influence individual behavior. In particular, how the different cultures in the Organizational Culture Assessment Instrument (OCAI) affect employees' main dimensions of Organizational Citizenship Behavior (OCB). A descriptive-confirmative ex post facto research was conducted by surveying 513 employees from over 150 organizations worldwide. The Kruskal-Wallis H-test was used to validate our model. The general hypothesis was confirmed, showing that the predominant organizational culture type affects the level and the kind of OCBs individuals demonstrate. It is possible to provide organizations with a breakdown of their employees' OCBs based on their OC type and which changes they can make to their organization's culture to increase the employees' OCB and, consequently, the efficiency of their organization.

Metal-Organic Frameworks Applications in Synergistic Cancer Photo-Immunotherapy.

Conventional cancer therapies, such as radiotherapy and chemotherapy, can have long-term side effects. Phototherapy has significant potential as a non-invasive alternative treatment with excellent selectivity. Nevertheless, its applicability is restricted by the availability of effective photosensitizers and photothermal agents, and its low efficacy when it comes to avoiding metastasis and tumor recurrence. Immunotherapy can promote systemic antitumoral immune responses, acting against metastasis and recurrence; however, it lacks the selectivity displayed by phototherapy, sometimes leading to adverse immune events. The use of metal-organic frameworks (MOFs) in the biomedical field has grown significantly in recent years. Due to their distinct properties, including their porous structure, large surface area, and inherent photo-responsive properties, MOFs can be particularly useful in the fields of cancer phototherapy and immunotherapy. MOF nanoplatforms have successfully demonstrated their ability to address several drawbacks associated with cancer phototherapy and immunotherapy, enabling an effective and low-side-effect combinatorial synergistical treatment for cancer. In the coming years, new advancements in MOFs, particularly regarding the development of highly stable multi-function MOF nanocomposites, may revolutionize the field of oncology.

Microplastic contamination in large migratory fishes collected in the open Atlantic Ocean.

Fishes are one of the most important components of the oceans and are exposed to several anthropogenic pressures, namely microplastic (MP), contaminants that are now ubiquitous worldwide. Taking advantage of the 2020 Circumnavigation Expedition carried by the NRP Sagres tall ship of the Portuguese Navy, fish samples from the southern Atlantic ocean were collected to evaluate possible MP contamination. In a total of 14 weeks of campaign, seven large migratory fishes of commercial interest were collected across the middle Atlantic Ocean and along the South American Atlantic coast. All individuals were contaminated with MPs, with an average of 18 ± 11 MPs/fish. In all fish sampled, both the gastrointestinal tract and gills presented MPs, indicating different contamination pathways including via their preys and from surrounding water, respectively. A total of 124 MPs were observed, where 72 % were fibers and 28 % particles, mostly of blue color (85 %), and with rayon and nylon as the most abundant polymers. This study is an important contribution to increase the scientific knowledge of MP contamination in mesopelagic fishes used for human consumption and collected in the open waters, reinforcing the need for further research regarding MP contamination in top predatory species from high trophic levels.

Validation of a Portuguese version of the Groningen radiotherapy-induced xerostomia questionnaire

Background: The aim of this study was to validate and determine at pretest level the reliability of the Portuguese version of the Groningen radiotherapy-induced xerostomia questionnaire. Material and methods: This study employed 37 head and neck cancer patients. Each patient signed an informed consent and responded to the Portuguese version of the questionnaire in the form of an interview. This was repeated again after 2 weeks. A standard single question provided a validity check. Data were analyzed using Cronbach's α to test its reliability and total and interitem correlation, and intraclass correlation to determine its internal consistency and test-retest reliability. Construct validity supported by objective measurements as salivary secretion was also investigated. Significance was set at .05. Results: Cronbach's α was 0.91 and 0.89 for the first and second test administrations, respectively, which indicates that the internal consistency was excellent. The intraclass correlation coefficient value for the test-retest reliability was 0.70. The correlation between the total score of the questionnaire and standard single dry mouth question was 0.72 for the first round, indicating a good correlation. Conclusions: Demonstrating very good psychometric properties, the Portuguese version of the Groningen radiotherapy-induced xerostomia questionnaire is a valid tool and can be considered a reliable instrument to measure xerostomia in head and neck cancer patients. (AU)

Bleaching efficacy and quality of life of different bleaching techniques - randomized controlled trial.

OBJECTIVES: To evaluate bleaching efficacy and oral health-related quality of life (ORHQoL) of three bleaching systems with similar hydrogen peroxide (HP) concentration for up to 6 months post-treatment. MATERIALS AND METHODS: A randomized controlled trial was designed with three parallel groups: group A - in-office 6% HP paint-on varnish; group B - at-home 6% HP with adaptable tray; group C - at-home 16% carbamide peroxide with custom tray. At three different stages (baseline, after bleaching, and 6-month follow-up), ORHQoL was evaluated by the OHIP-14 questionnaire and tooth color of the upper canines and central incisors were measured by two shade guides and a spectrophotometer (measuring CIE L*a*b* with respective color/whiteness differences - ΔE00/ΔWID). Results were presented as mean and 95% confidence intervals and statistical tests were performed appropriately, considering a significance level of α = 0.05. RESULTS: All groups presented significant color differences (P < 0.05) between all stages, with ΔE00/ΔWID surpassing the perceptibility threshold in 98% cases, with group C's results being significantly (P < 0.05) higher when compared to other groups, although with significantly (P < 0.05) higher values of color relapse. Significative ORHQoL improvements (P < 0.05) were detected after bleaching in a global analysis with no differences between techniques. CONCLUSIONS: All techniques presented bleaching efficacy, color stability, and improvements in ORHQoL up to 6 months post-treatment. CLINICAL SIGNIFICANCE: Clinicians may consider both at-home and in-office bleaching techniques with 6% HP to attain long-lasting satisfactory clinical results while producing positive changes in ORHQoL.

3D Bioprinting: An Enabling Technology to Understand Melanoma.

Melanoma is a potentially fatal cancer with rising incidence over the last 50 years, associated with enhanced sun exposure and ultraviolet radiation. Its incidence is highest in people of European descent and the ageing population. There are multiple clinical and epidemiological variables affecting melanoma incidence and mortality, such as sex, ethnicity, UV exposure, anatomic site, and age. Although survival has improved in recent years due to advances in targeted and immunotherapies, new understanding of melanoma biology and disease progression is vital to improving clinical outcomes. Efforts to develop three-dimensional human skin equivalent models using biofabrication techniques, such as bioprinting, promise to deliver a better understanding of the complexity of melanoma and associated risk factors. These 3D skin models can be used as a platform for patient specific models and testing therapeutics.

Fabrication of Polymer/Graphene Biocomposites for Tissue Engineering.

Graphene-based materials (GBM) are considered one of the 21st century's most promising materials, as they are incredibly light, strong, thin and have remarkable electrical and thermal properties. As a result, over the past decade, their combination with a diverse range of synthetic polymers has been explored in tissue engineering (TE) and regenerative medicine (RM). In addition, a wide range of methods for fabricating polymer/GBM scaffolds have been reported. This review provides an overview of the most recent advances in polymer/GBM composite development and fabrication, focusing on methods such as electrospinning and additive manufacturing (AM). As a future outlook, this work stresses the need for more in vivo studies to validate polymer/GBM composite scaffolds for TE applications, and gives insight on their fabrication by state-of-the-art processing technologies.

Hydrogen Peroxide Release Kinetics of Four Tooth Whitening Products-In Vitro Study.

Tooth whitening efficacy can be influenced by several factors, of which concentration and application time are two of the most important. This in vitro study aimed to evaluate the initial content and release kinetics of the hydrogen peroxide (HP) content, or the carbamide peroxide (CP) content as converted to its HP equivalent, of four tooth whitening products with different concentrations (6% HP, 16% CP, 10% CP, and 5% CP). Titrations with Cerium Sulphate IV were performed to determine HP concentration. HP release kinetics were evaluated by a spectrophotometric technique. The results were expressed as the mean values and 95% confidence interval of the percentage of hydrogen peroxide content during release kinetics. One sample t-test, one-way ANOVA, Tukey post hoc testing, and Pearson correlation testing were used, as appropriate, with a significance level of α = 0.05. The concentration of titrated HP was higher than that indicated by the manufacturers in all tested products (p < 0.01). At the minimum application times indicated by the manufacturers, all products released at least 85% of HP content; the gel containing 10% CP registered the lowest release at 85.49 (81.52-89.46). There was a significant HP release in all products during the application times indicated by the manufacturers. Further studies are needed to assess in vitro release kinetics.

The individuals' discretionary behaviors at work. An overview and analysis of its growing interest.

Nearly forty years have passed since the term Organizational Citizenship Behavior appeared. Despite a current consensus among scholars about the citizenship gestures as prosocial acts of employees that benefit the organization, it does not apply commonly to the exponential growth of all OCB-related concepts. The concept's expansion has confused the researchers and practitioners, mainly when choosing the most appropriate instruments (constructs) and dimensions to use in their area of interest and context. A systematic literature review was conducted and 420 articles were analyzed. Results point that the trends on OCB-like behaviors goes higher, with an average annual growth rate of new studies of 3.13%. United States (39%) and China (25%) lead but some "under-studied" contexts like Africa, Latin America, the Caribbean, and Oceania start being studied towards OCB. New trends appear in the spectrum of the many different domains and disciplines related to OCB, while leadership and task performance remain the most studied domains and disciplines. In future studies, researchers must freely choose the OCB dimensions and constructs they want to use or adapt to meet their needs and research needs since there is no written rule about their use, only the care to be taken with the context and discipline studying.

An injectable, dual crosslinkable hybrid pectin methacrylate (PECMA)/gelatin methacryloyl (GelMA) hydrogel for skin hemostasis applications.

Biomaterials for effective hemorrhage control are urgently needed in clinics as uncontrolled bleeding is associated with high mortality. Herein, we developed an injectable and in situ photo-crosslinkable hybrid hemostatic hydrogel by combining pectin methacrylate (PECMA) and gelatin methacryloyl (GelMA). This modular material system combines ionic- and photo-crosslinking chemistries to design interpenetrating networks (IPN) exhibiting tunable rheology, highly porous structure, and controllable swelling and mechanical properties. By simply changing the calcium (0-15 mM) and polymer (1.5-7%) content used for the sequential crosslinking of hydrogels via calcium gelation and UV-photopolymerization, it was possible to precisely modulate the injectability, degradation, and swelling ratio. Moreover, it is demonstrated that PECMA/GelMA hydrogels present good cytocompatibility and uniquely synergize the hemostatic properties of calcium ions on PECMA, the amine residues on GelMA, and the highly porous network toward rapid blood absorption and fast coagulation effect. An in vitro porcine skin bleeding model confirmed that the hydrogel could be directly injected into the wound and rapidly photo-crosslinked, circumventing the bleeding and decreasing the coagulation time by 39%. Importantly, the crosslinked hydrogel could be easily removed to prevent secondary wound injury. Overall, this injectable hybrid PECMA/GelMA hydrogel stands as a promising hemostatic material.

Engineering Modular Half-Antibody Conjugated Nanoparticles for Targeting CD44v6-Expressing Cancer Cells.

Gastric cancer (GC) remains a major cause of death worldwide mainly because of the late detection in advanced stage. Recently, we proposed CD44v6 as a relevant marker for early detection of GC, opening new avenues for GC-targeted theranostics. Here, we designed a modular nanoscale system that selectively targets CD44v6-expressing GC cells by the site-oriented conjugation of a new-engineered CD44v6 half-antibody fragment to maleimide-modified polystyrene nanoparticles (PNPs) via an efficient bioorthogonal thiol-Michael addition click chemistry. PNPs with optimal particle size (200 nm) for crossing a developed biomimetic CD44v6-associated GC stromal model were further modified with a heterobifunctional maleimide crosslinker and click conjugated to the novel CD44v6 half-antibody fragment, obtained by chemical reduction of full antibody, without affecting its bioactivity. Collectively, our results confirmed the specific targeting ability of CD44v6-PNPs to CD44v6-expressing cells (1.65-fold higher than controls), highlighting the potential of CD44v6 half-antibody conjugated nanoparticles as promising and clinically relevant tools for the early diagnosis and therapy of GC. Additionally, the rational design of our nanoscale system may be explored for the development of several other nanotechnology-based disease-targeted approaches.

Tissue-specific engineering: 3D bioprinting in regenerative medicine.

Despite its complexity, the human body is composed of only four basic tissue types, namely epithelial, connective, muscular and nervous tissues. Notably, each tissue is an assemblage of similarly functional cells united in performing a specific function. Instead of mimicking functionality mechanically, three-dimensional (3D) bioprinting based on histological categories is a strategy designed with multiple materials and techniques, which is a versatile technology able to form functional organ structures in line with simplicity. This review aims to provide an overview of tissue-specific 3D bioprinting based on the biological characteristics of four tissue types, including the histological features, biomaterials and corresponding applications. It first briefly introduces the goals of tissue-specific bioprinting and then summarizes the major techniques and identification of particular material development. Moreover, its remarkable regenerative power in replacement therapy and novel outbreak in particular tissues are assembled by epithelial, connective, nerve and muscle tissues. Finally, we discuss challenges and future prospects of tissue-specific based 3D bioprinting in biomedicine, hoping to further inspire the development.

3D Cell Culture Models as Recapitulators of the Tumor Microenvironment for the Screening of Anti-Cancer Drugs.

Today, innovative three-dimensional (3D) cell culture models have been proposed as viable and biomimetic alternatives for initial drug screening, allowing the improvement of the efficiency of drug development. These models are gaining popularity, given their ability to reproduce key aspects of the tumor microenvironment, concerning the 3D tumor architecture as well as the interactions of tumor cells with the extracellular matrix and surrounding non-tumor cells. The development of accurate 3D models may become beneficial to decrease the use of laboratory animals in scientific research, in accordance with the European Union's regulation on the 3R rule (Replacement, Reduction, Refinement). This review focuses on the impact of 3D cell culture models on cancer research, discussing their advantages, limitations, and compatibility with high-throughput screenings and automated systems. An insight is also given on the adequacy of the available readouts for the interpretation of the data obtained from the 3D cell culture models. Importantly, we also emphasize the need for the incorporation of additional and complementary microenvironment elements on the design of 3D cell culture models, towards improved predictive value of drug efficacy.