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Abstract The impact of sodium hexametaphosphate (SHMP) on the aggregation behavior of α-lactalbumin (α-LA) was studied at pH 7.4 and 2.0. Turbidity measurements showed a concentration-dependent aggregation of α-LA at pH 2.0 in the presence of SHMP, while no aggregation was observed at pH 7.4. Light scattering (LS) and Thioflavin-T (ThT) data revealed that the aggregation was rapid, following nucleation-independent pathways. In other kinetics experiments such as turbidity and ThT confirmed that SHMP-induced α-LA aggregation was dependent on SHMP concentration rather than incubation time. Once formed, the aggregates remained unchanged for up to five days. Intrinsic fluorescence studies indicated conformational changes in α-LA upon SHMP addition, and dye-binding assays with ThT and Congo Red demonstrated the formation of amyloid-like aggregates. Far-UV circular dichroism (CD) data suggested a structural transition from α-helical to β-structures in α-LA in the presence of SHMP at pH 2.0. Molecular docking studies confirmed stronger interactions between α-LA and SHMP at pH 2.0 (ΔG = −6.2 kcal/mol) compared to pH 7.4 (ΔG = −5.3 kcal/mol), driven by electrostatic forces and hydrogen bonding. These results suggest that SHMP induces amyloid-like aggregation of α-LA, particularly at acidic pH.

Aims: Distal femoral osteotomies (DFOs) are commonly used for the correction of valgus deformities and lateral compartment osteoarthritis. However, the impact of a DFO on subsequent total knee arthroplasty (TKA) function remains a subject of debate. Therefore, the purpose of this study was to determine the effect of a unilateral DFO on subsequent TKA function in patients with bilateral TKAs, using the contralateral knee as a self-matched control group. Methods: The inclusion criteria consisted of patients who underwent simultaneous or staged bilateral TKA after prior unilateral DFO between 1972 and 2023. The type of osteotomy performed, osteotomy hardware fixation, implanted TKA components, and revision rates were recorded. Postoperative outcomes including the Forgotten Joint Score-12 (FJS-12), Tegner Activity Scale score, and subjective knee preference were also obtained at final follow-up. Results: A total of 21 patients underwent bilateral TKA following unilateral DFO and were followed for a mean of 31.5 years (SD 11.1; 20.2 to 74.2) after DFO. The mean time from DFO to TKA conversion was 13.1 years (SD 9.7) with 13 (61.9%) of DFO knees converting to TKA more than ten years after DFO. There was no difference in arthroplasty implant systems employed in both the DFO-TKA and TKA-only knees (p > 0.999). At final follow-up, the mean FJS-12 of the DFO-TKA knee was 62.7 (SD 36.6), while for the TKA-only knee it was 65.6 (SD 34.7) (p = 0.328). In all, 80% of patients had no subjective knee preference or preferred their DFO-TKA knee. Three DFO-TKA knees and two TKA-only knees underwent subsequent revision following index arthroplasty at a mean of 12.8 years (SD 6.9) and 8.5 years (SD 3.8), respectively (p > 0.999). Conclusion: In this self-matched study, DFOs did not affect subsequent TKA function as clinical outcomes, subjective knee preference, and revision rates were similar in both the DFO-TKA and TKA-only knees at mean 32-year follow-up. Cite this article: Bone Jt Open 2024;5(11):1013–1019.

Abstract Ficus auriculata (Timla) is an underutilized wild fruit compared to the more commonly consumed fruit of the same genus, Ficus carica (Anjeer). This review focuses on the industrial and pharmaceutical benefits of F. auriculata while highlighting its potential as a valuable alternative to F. carica. Based on the vast literature found in Scopus database, both fruits have significant nutritional and medicinal value; however, due to the rarity and lack of widespread knowledge, F. auriculata still remains underutilized, unlike F. carica, which is well known and more popular in the human diet. In terms of nutrition, F. auriculata is superior in various aspects like protein (3–5.32%), magnesium (68.0 mg/100 g), potassium (329.0 mg/100 g), and iron (5.0 mg/100 g). As a result, it plays a pivotal role for addressing nutrient deficiencies. Besides, it has rich history of many noteworthy applications in traditional remedies which are also recognized in modern pharmaceutical research. This could be attributed to the presence of phytocompounds. Ficus auriculata needs to be promoted, not only as a nutritional addition to the plate but also as an economically viable fruit. Based on its considerable potential, it is possible that F. auriculata can also be used as an alternative to F. carica; however, further research and promotion of the benefits of F. auriculata could help to increase its utilization and provide additional health and economic benefits to communities. Moreover, omics technologies based comprehensive study of fruits can assist in the identification of bioactive metabolites, nutritional content, and possible health benefits. It offers insights into the genetic composition, gene expression patterns, protein activities, and metabolic pathways of fruits thereby revealing the mechanisms driving flavor, color, and disease resistance.

Abstract The emergence of antimicrobial resistance has created an urgent need for alternative treatments against bacterial pathogens. Here, we investigated kinase inhibitors as potential host-directed therapies (HDTs) against intracellular bacteria, specifically Salmonella Typhimurium (Stm) and Mycobacterium tuberculosis (Mtb). We screened 827 ATP-competitive kinase inhibitors with known target profiles from two Published Kinase Inhibitor Sets (PKIS1 and PKIS2) using intracellular infection models for Stm and Mtb, based on human cell lines and primary macrophages. Additionally, the in vivo safety and efficacy of the compounds were assessed using zebrafish embryo infection models. Our screen identified 11 hit compounds for Stm and 17 hit compounds for Mtb that were effective against intracellular bacteria and non-toxic for host cells. Further experiments were conducted to prioritize Stm hit compounds that were able to clear the intracellular infection in primary human macrophages. From these, two structurally related Stm hit compounds, GSK1379738A and GSK1379760A, exhibited significant activity against Stm in infected zebrafish embryos. In addition, we identified compounds that were active against intracellular Mtb, including morpholino-imidazo/triazolo-pyrimidinones that target PIK3CB, as well as 2-aminobenzimidazoles targeting ABL1. Overall, this study provided insights into kinase targets acting at the host–pathogen interface and identified several kinase inhibitors as potential HDTs.

Wood hemicelluloses, specifically galactoglucomannans (GGM) and glucuronoxylans (GX) are investigated for their efficacy in spray-dried microencapsulation of Saccharomyces cerevisiae subsp. boulardii (SB). This study demonstrated that GGM and GX effectively protected SB during spray drying at feed concentrations of 15 and 20 % and inlet air temperatures of 105 and 140 °C, ensuring survival rates over 90 %, comparable to gum Arabic, with all achieving over 108 cfu/g in the microcapsules. However, GGM and GX were unable to sustain SB viability when the microcapsules were stored at 33 and 75 % RH (22 °C), beyond 21 and 7 days, respectively. When stored at 4 °C, GX demonstrated a greater ability to protect SB than GGM, with log-cycle reductions of 3 and 6, respectively, after two months. Microstructure analyses showed almost all SB were entrapped by wall materials, with many microcapsules having a wall thickness of 1 − 2 µm. Overall, GX is effective for stable yeast probiotic powders, enabling the development of new probiotic enhanced formulations with prolonged viability and stability.

Unstable osteochondritis dissecans lesions of the medial femoral condyle have classically been treated with open reduction and fixation under direct visualization through an open arthrotomy. Given the value of avoiding open arthrotomies, we present an arthroscopic approach for lesion elevation, debridement, and fixation. The lesion is first elevated using an arthroscopic elevator, leaving a laterally based osseous hinge. Once elevated, fibrous debris is debrided from the base of the lesion. Subsequently, the fragment is reduced, and percutaneous transpatellar instrumentation is used for fixation. The use of this technique allows for excellent mobilization, debridement, and fixation of the osteochondritis dissecans lesion while minimizing violation of periarticular soft tissues.

Biosynthesizing conjugated linoleic acid (CLA) using food processing by-products offers a practical and efficient method for producing this valuable compound, delivering health benefits, promoting sustainability, and providing economic advantages. Here, we optimized the fermentation conditions for in situ biosynthesis of CLA using Bacillus coagulans IBRC-M 10807, with grapeseed oil by-product providing the linoleic acid precursor, focusing on temperature, pH, incubation time, concentrations of grapeseed oil and yeast extract. The total CLA was measured using a UV–Vis spectrophotometer at 233 nm, and the concentration was evaluated using a standard curve of conjugated linoleic acid. To analyze the isomers and identify the functional groups of biosynthesized CLA, gas chromatography (GC) and Fourier-transform infrared spectroscopy (FTIR) were used, respectively. The results indicated that the maximum amount of CLA production was 225.37 ± 0.74 μg/mL, obtained under optimal conditions: an initial pH of 6.8, a temperature of 44.8°C, an incubation time of 48 h, a grapeseed oil concentration of 1,000 μg/mL, and a yeast extract concentration of 3.88%. Characterization of the produced CLA revealed two main isomers—CLA 1: c-9, t-11 C18:2, and CLA 2: t-10, c-12 C18:2 methyl esters. These results suggest that B. coagulans has high potential to produce CLA as a bioactive compound, offering an excellent opportunity for use in various industries, including nutraceuticals, foods, pharmaceuticals, cosmetics, and animal feeds.

Objectives: To evaluate if race is associated with disparities in receipt of radiation (RT) and outcomes for Medicare patients with cervical cancer who are candidates for primary radiation-chemotherapy. Methods: This SEER-Medicare retrospective study included White and Black patients with stage IB1 through IVA squamous cell carcinoma or adenocarcinoma diagnosed 2000–2017 who were candidates for primary radiation-chemotherapy. Receipt of treatment by race and associated cancer specific (CSS) and overall survival (OS) outcomes were analyzed using frequency distributions, chi squared, log rank, multivariable Cox proportional-hazards models, and multivariable logistic models. Results: 1038 patients (84.9 % White and 15.1 % Black) were included. 825 (79.5 %) received RT, and 601 (57.9 %) received brachytherapy (BT). Blacks were more likely to undergo RT than Whites (86.0 % vs. 78.3 %, p = 0.028) and had similar rates of BT (58.0 % vs. 57.9 %, p = 0.986). Median RT duration was 64.0 days (IQR 52.0, 75.0), and 276 (33.5 %) completed treatment in ≤ 56 days, with no differences by race (p = 0.488, 0.303, respectively). BT was more frequently provided at larger hospitals, National Cancer Institute-designated cancer centers, and teaching hospitals. When adjusted for covariates, no significant differences in RT, BT, or RT duration by race were identified. Median unadjusted OS was 3.58 years (95 % CI 2.92, 4.42) for White patients and 2.50 years (95 % CI 2.0, 5.25) for Black patients, with no differences in OS (HR 0.93, 95 % CI 0.75, 1.13) or CSS (HR 1.13, 95 %CI 0.86, 1.43). Conclusions: Black Medicare patients with cervical cancer had greater receipt of RT than White patients, similar rates of BT, and no difference in survival.

Previous theoretical frameworks used to research and explain creative design processes tend to privilege individual expression and not address the context in which the process occurs. This is problematic due to the ways in which creative activities are embedded in and shaped by socio-cultural and historic contexts. In this article we focus on the ways in which cultural historic activity theory (CHAT) and its analytical activity systems can be used to reveal creative processes in context. We draw on the tools and concepts of CHAT and data from a study of visual communication design (VCD) students and lecturers situated within a transnational higher education (TNHE) context at a university in China. An analytical framework was constructed to research practices used in creative design process learning. The dynamic nature of CHAT offers design process research methods a set of analytical tools to capture the powerful parts played by artifacts and interactions within specific sociocultural and historic learning contexts.

Introduction: Articular cartilage injuries of the knee are a complex and challenging clinical pathology. Objectives: The purpose of this study was to establish consensus statements via a Delphi process on cartilage fixation techniques and indications. Methods: A consensus process on knee cartilage injuries utilizing a modified Delphi technique was conducted. Seventy-nine surgeons across 17 countries participated in these consensus statements. Fourteen questions were generated on cartilage fixation, with 3 rounds of questionnaires and final voting occurring. Consensus was defined as achieving 80% to 89% agreement, whereas strong consensus was defined as 90% to 99% agreement, and unanimous consensus was defined as 100% agreement with a proposed statement. Results: Of the 14 total questions and consensus statements on cartilage fixation developed from 3 rounds of voting, 0 achieved unanimous consensus, 8 achieved strong consensus, 3 achieved consensus, and 3 did not achieve consensus. Conclusions: The statements achieving strong consensus related to indications and contraindications, the expectation that younger patients experience better outcomes, and an agreement that procedures should be done as soon as possible but no limit on timeframe, cartilage preparation techniques, and the use of fragment stability to guide cartilage fixation techniques. The statements that did not achieve consensus related to the minimum size of cartilage fragments that can be fixed, if purely cartilaginous lesions can be fixed, and the fixation techniques.

ABSTRACT Mycobacterium tuberculosis (Mtb) as well as nontuberculous mycobacteria are intracellular pathogens whose treatment is extensive and increasingly impaired due to the rise of mycobacterial drug resistance. The loss of antibiotic efficacy has raised interest in the identification of host-directed therapeutics (HDT) to develop novel treatment strategies for mycobacterial infections. In this study, we identified amiodarone as a potential HDT candidate that inhibited both intracellular Mtb and Mycobacterium avium in primary human macrophages without directly impairing bacterial growth, thereby confirming that amiodarone acts in a host-mediated manner. Moreover, amiodarone induced the formation of (auto)phagosomes and enhanced autophagic targeting of mycobacteria in macrophages. The induction of autophagy by amiodarone is likely due to enhanced transcriptional regulation, as the nuclear intensity of the transcription factor EB, the master regulator of autophagy and lysosomal biogenesis, was strongly increased. Furthermore, blocking lysosomal degradation with bafilomycin impaired the host-beneficial effect of amiodarone. Finally, amiodarone induced autophagy and reduced bacterial burden in a zebrafish embryo model of tuberculosis, thereby confirming the HDT activity of amiodarone in vivo. In conclusion, we have identified amiodarone as an autophagy-inducing antimycobacterial HDT that improves host control of mycobacterial infections.IMPORTANCEDue to the global rise in antibiotic resistance, there is a strong need for alternative treatment strategies against intracellular bacterial infections, including Mycobacterium tuberculosis (Mtb) and non-tuberculous mycobacteria. Stimulating host defense mechanisms by host‐directed therapy (HDT) is a promising approach for treating mycobacterial infections. This study identified amiodarone, an antiarrhythmic agent, as a potential HDT candidate that inhibits the survival of Mtb and Mycobacterium avium in primary human macrophages. The antimycobacterial effect of amiodarone was confirmed in an in vivo tuberculosis model based on Mycobacterium marinum infection of zebrafish embryos. Furthermore, amiodarone induced autophagy and inhibition of the autophagic flux effectively impaired the host-protective effect of amiodarone, supporting that activation of the host (auto)phagolysosomal pathway is essential for the mechanism of action of amiodarone. In conclusion, we have identified amiodarone as an autophagy-inducing HDT that improves host control of a wide range of mycobacteria.

BackgroundThe occurrence of peritoneal metastasis (PM) in patients with colorectal cancer (CRC) has a dismal prognosis. There is often limited response to systemic- and immunotherapy, even in microsatellite unstable (MSI) CRC. To overcome therapy resistance, it is critical to understand local immune environment in the peritoneal cavity, and to develop models to study anti-tumor immune responses. Here, we defined the peritoneal immune system (PerIS) in PM-CRC patients and evaluate the pre-clinical potential of a humanized immune system (HIS) mouse model for PM-CRC.MethodsWe studied the human PerIS in PM-CRC patients (n=20; MSS 19/20; 95%) and in healthy controls (n=3). HIS mice (NODscid gamma background; n=18) were generated, followed by intraperitoneal injection of either saline (HIS control; n=3) or human MSS/MSI CRC cell lines HUTU80, MDST8 and HCT116 (HIS-PM, n=15). Immune cells in peritoneal fluid and peritoneal tumors were analyzed using cytometry by time of flight (CyTOF).ResultsThe human and HIS mouse homeostatic PerIS was equally populated by NK cells and CD4+- and CD8+ T cells, however differences were observed in macrophage and B cell abundance. In HIS mice, successful peritoneal engraftment of both MSI and MSS tumors was observed (15/15; 100%). Both in human PM-CRC and in the HIS mouse PM-CRC model, we observed that MSS PM-CRC triggered a CD4+ Treg response in the PerIS, while MSI PM-CRC drives CD8+ TEMs responses.ConclusionIn conclusion, T cell responses in PM-CRC in HIS mice mirror those in human PM-CRC, making this model suitable to study antitumor T cell responses in PM-CRC.

Background Tertiary lymphoid structures (TLSs) are thought to stimulate antitumor immunity and positively impact prognosis and response to immune checkpoint blockade. In gastric cancers (GCs), however, TLSs are predominantly found in GC with poor prognosis and limited treatment response. We, therefore, hypothesize that immune cell composition and function of TLS depends on tumor location and the tumor immune environment.Methods Spatial transcriptomics and immunohistochemistry were used to characterize the phenotype of CD45+ immune cells inside and outside of TLS using archival resection specimens from GC primary tumors and peritoneal metastases.Results We identified significant intrapatient and interpatient diversity of the cellular composition and maturation status of TLS in GC. Tumor location (primary vs metastatic site) accounted for the majority of differences in TLS maturity, as TLS in peritoneal metastases were predominantly immature. This was associated with higher levels of tumor-infiltrating macrophages and Tregs and less plasma cells compared with tumors with mature TLS. Furthermore, mature TLSs were characterized by overexpression of antitumor immune pathways such as B cell-related pathways, MHC class II antigen presentation while immature TLS were associated with protumor pathways, including T cell exhaustion and enhancement of DNA repair pathways in the corresponding cancer.Conclusion The observation that GC-derived peritoneal metastases often contain immature TLS which are associated with immune suppressive regulatory tumor-infiltrating leucocytes, is in keeping with the lack of response to immune checkpoint blockade and the poor prognostic features of peritoneal metastatic GC, which needs to be taken into account when optimizing immunomodulatory strategies for metastatic GC.

Abstract With the huge progress in micro-electronics and artificial intelligence, the ultrasound probe has become the bottleneck in further adoption of ultrasound beyond the clinical setting (e.g. home and monitoring applications). Today, ultrasound transducers have a small aperture, are bulky, contain lead and are expensive to fabricate. Furthermore, they are rigid, which limits their integration into flexible skin patches. New ways to fabricate flexible ultrasound patches have therefore attracted much attention recently. First prototypes typically use the same lead-containing piezo-electric materials, and are made using micro-assembly of rigid active components on plastic or rubber-like substrates. We present an ultrasound transducer-on-foil technology based on thermal embossing of a piezoelectric polymer. High-quality two-dimensional ultrasound images of a tissue mimicking phantom are obtained. Mechanical flexibility and effective area scalability of the transducer are demonstrated by functional integration into an endoscope probe with a small radius of 3 mm and a large area (91.2×14 mm2) non-invasive blood pressure sensor.