The genetic signature left by the range expansion of red foxes in Australia is detectable after more than 80 years of population stability.

Reconstructing biological invasions from historical sources can provide insights into how they occur but are difficult to do when invasions are poorly documented. Genetic signatures left by invaders can also offer insights into invasion routes, points of origin and general biology but often present conclusions that are contradictory to expectations. Here, we test the ability of continental-wide microsatellite genotype data from 29 loci and 3122 samples to reconstruct the well-documented invasion of red foxes Vulpes vulpes from the United Kingdom into Australia over 150 years ago, an invasion that has led to the extinction of many native species. Our analysis reveals several key signals of invasion evident in Australian foxes. They display lower levels of diversity than foxes sampled from the UK, exhibit clines in diversity from the point of introduction (south-east Australia) to the edge of their range, and show strong evidence of allele surfing in westerly and north-easterly directions. These characteristics are consistent with a single point of origin followed by rapid expansion in westerly and north-easterly directions as suggested by historical records. We also find little genetic structure in foxes across Australia with only the vast Nullarbor Plains and Great Victoria Desert region presenting a detectable barrier to their dispersal. As such, no mainland region within the current range of foxes can be considered genetically isolated and therefore appropriate for localised eradication efforts. Overall, our analyses demonstrate the ability of comprehensive population genetic studies to reconstruct invasion histories even after more than 80 years since colonisation was stabilised.

Genetic Modification of Tumor-Infiltrating Lymphocytes, Peripheral T Cells, and T-Cell Model Cell Lines.

Genetic modification of tumor-infiltrating lymphocytes (TILs) or circulating T cells has become an important avenue in cancer therapy. Here we describe a comprehensive method for establishing and expanding TIL cultures and genetically modifying them with a gene of interest (GOI) via retroviral transduction or mRNA transfection. The method includes all the important steps starting with TIL extraction from tumors through to the maintenance of the genetically modified TILs. The protocol includes instructions for retroviral transduction and mRNA transfection of circulating T cells or T-cell lines. The GOIs most commonly introduced into the target cells are chimeric antigen receptors (CARs); genetic adjuvants, such as membrane-bound interleukins; and antitumor T-cell receptors (TCRs).

Dento-skeletal Sequel of Hyrax in Nonsyndromic Cleft Lip and Palate: A Systematic Review and Meta-analysis.

Aim: To critically evaluate and review the dento-skeletal outcomes of hyrax with or without nonsurgical protraction therapy in nonsyndromic cleft lip and palate (CLP) patients. Materials and methods: Electronic databases were searched from PubMed, Cochrane, Scopus, SciELO, Lilacs, and Google Scholar from January 2005 to April 2022 to identify studies reporting on the dento-skeletal effect of hyrax with or without nonsurgical protraction therapy among children aged 7-15 years with CLP before the secondary alveolar bone grafting (SABG) procedure. Abstracts and subsequently eligible full-text articles were screened. The risk of bias in the included randomized and nonrandomized studies was assessed using Cochrane collaboration's risk of bias tool. The random-effects meta-analyses of the mean difference were carried out. Results: A total of 423 studies were identified. After a thorough screening, 19 studies met the inclusion criteria. Of these 19 studies, 13 studies were conducted with only hyrax appliances, six studies had hyrax with nonsurgical protraction therapy. Conclusion: Hyrax had increased intermolar width, anterior nasal cavity width, and posterior maxillary width compared to inverted mini hyrax with p-values = 0.027, 0.004, and 0.03, respectively. Inverted mini hyrax had more canine inclination on noncleft than hyrax. Hyrax and fan-type expanders had an equivalent effect on the dento-skeletal structures indicating that hyrax can be used as an alternative to fan-type expanders. Hyrax with protraction therapy had a remarkable change in ANB angle compared to noncleft patients. How to cite this article: Deepika U, Ray P, Nayak A, et al. Dento-skeletal Sequel of Hyrax in Nonsyndromic Cleft Lip and Palate: A Systematic Review and Meta-analysis. Int J Clin Pediatr Dent 2023;16(6):882-896.

Isolation and Maintenance of Tumor-Infiltrating Lymphocytes for Translational and Clinical Applications: Established Methods and New Developments.

Adoptive cell transfer (ACT) of in vitro expanded tumor-infiltrating lymphocytes (TIL) for the treatment of patients with advanced stages of metastatic melanoma remains one of the most beneficial therapies eliciting long-lasting responses. Methods and protocols used to expand TIL have evolved over time, utilizing different culture devices and other tools, to streamline and maximize the end product in both numbers and quality. Summarized in this chapter are the latest protocols used in the TIL program at MDACC.

Formation of Polymer-Carbon Nanotube Composites by Two-Step Supercritical Fluid Treatment.

An approach for polymer-carbon nanotube (CNT) composite preparation is proposed based on a two-step supercritical fluid treatment. The first step, rapid expansion of a suspension (RESS) of CNTs in supercritical carbon dioxide, is used to de-bundle CNTs in order to simplify their mixing with polymer in solution. The ability of RESS pre-treatment to de-bundle CNTs and to cause significant bulk volume expansion is demonstrated. The second step is the formation of polymer-CNT composite from solution via supercritical antisolvent (SAS) precipitation. SAS treatment allows avoiding CNT agglomeration during transition from a solution into solid state due to the high speed of phase transition. The combination of these two supercritical fluid methods allowed obtaining a polycarbonate-multiwalled carbon nanotube composite with tensile strength two times higher compared to the initial polymer and enhanced elasticity.

Pharmaceutical amorphous solid dispersion: A review of manufacturing strategies.

Amorphous solid dispersions (ASDs) are popular for enhancing the solubility and bioavailability of poorly water-soluble drugs. Various approaches have been employed to produce ASDs and novel techniques are emerging. This review provides an updated overview of manufacturing techniques for preparing ASDs. As physical stability is a critical quality attribute for ASD, the impact of formulation, equipment, and process variables, together with the downstream processing on physical stability of ASDs have been discussed. Selection strategies are proposed to identify suitable manufacturing methods, which may aid in the development of ASDs with satisfactory physical stability.

Successful Prophylactic Endovascular Therapy for a Rapidly Expanding Hepatic Arterial Aneurysm in a Patient with Vascular Ehlers-Danlos Syndrome.

Vascular Ehlers-Danlos syndrome (vEDS) causes fatal vascular complications due to vascular fragility. However, invasive therapeutic procedures are generally avoided except in emergencies. We report a case of vEDS presenting with rapid expansion of a hepatic arterial aneurysm successfully treated using prophylactic endovascular therapy. A 43-year-old woman with vEDS confirmed by genetic testing was hospitalized for a symptomatic hepatic arterial aneurysm that expanded rapidly within a week. Prophylactic coil embolization was then successfully performed. Although the general applicability of this approach cannot be determined, prophylactic endovascular therapy can clearly be an option for arterial aneurysms at high risk of rupture.

The effect of cetirizine, a histamine 1 receptor antagonist, on bone remodeling after calvarial suture expansion.

OBJECTIVE: The objective of this study was to evaluate the effects of cetirizine, a histamine 1 receptor antagonist, on bone remodeling after calvarial suture expansion. METHODS: Sixty male Sprague-Dawley rats were divided into 4 groups; the phosphate-buffered saline (PBS)-injected no expansion group, cetirizine-injected no expansion group, PBS-injected expansion group, and cetirizine-injected expansion group, and were observed at 7, 14, and 28 days. Five rats per group were examined at each observation day. Daily injections of cetirizine or PBS were administered to the relevant groups starting 2 weeks prior to expander insertion. A rapid expander was inserted in the calvarial bone to deliver 100 cN of force to the parietal suture. The specimens were prepared for hematoxylin and eosin and tartrate-resistant acid phosphatase (TRAP) staining. Suture opening and bone regeneration were evaluated using microcomputed tomography and bone histomorphometric analysis. Serum blood levels of osteocalcin and carboxy-terminal collagen crosslinks (CTX) were also evaluated. RESULTS: TRAP-positive cell counts and CTX levels decreased while osteocalcin levels increased in the cetirizine-injected expansion group at observation day 28. In the expansion groups, the mineralized area gradually increased throughout the observation period. At day 28, the cetirizine-injected expansion group showed greater bone volume density, greater mineralized area, and narrower average suture width than did the PBS-injected expansion group. CONCLUSIONS: Cetirizine injection facilitated bone formation after suture expansion, mostly by suppressing osteoclastic activity. Histamine 1 receptor antagonists may aid in bone formation after calvarial suture expansion in the rat model.

Genetic Modification of Tumor-Infiltrating Lymphocytes via Retroviral Transduction.

Adoptive T cell therapy (ACT) holds great promise for cancer treatment. One approach, which has regained wide interest in recent years, employs antitumor T cells isolated from tumor lesions ("tumor-infiltrating lymphocytes" or TIL). It is now appreciated that a considerable proportion of anti-melanoma TIL recognize new HLA-binding peptides resulting from somatic mutations, which occurred during tumor progression. The clinical efficacy of TIL can potentially be improved via their genetic modification, designed to enhance their survival, homing capacity, resistance to suppression, tumor killing ability and additional properties of clinical relevance. Successful implementation of such gene-based strategies critically depends on efficient and reproducible protocols for gene delivery into clinical TIL preparations. Here we describe an optimized protocol for the retroviral transduction of TIL. As the experimental system we employed anti-melanoma TIL cultures prepared from four patients, recombinant retrovirus encoding an anti-CD19 chimeric antigen receptor (CAR) as a model gene of interest and CD19+ and CD19- human cell lines serving as target cells. Transduction on day 7 of the rapid expansion protocol (REP) resulted in 69 ± 8% CAR positive TIL. Transduced, but not untransduced TIL, from the four patients responded robustly to CD19+, but not CD19- cell lines, as judged by substantial secretion of IFN-γ following co-culture. In light of the rekindled interest in antitumor TIL, this protocol can be incorporated into a broad range of gene-based approaches for improving the in-vivo survival and functionality of TIL in the clinical setting.

Self-Delivering RNAi Targeting PD-1 Improves Tumor-Specific T Cell Functionality for Adoptive Cell Therapy of Malignant Melanoma.

Adoptive cell therapy (ACT) is becoming a prominent alternative therapeutic treatment for cancer patients relapsing on traditional therapies. In parallel, antibodies targeting immune checkpoint molecules, such as cytotoxic-T-lymphocyte-associated antigen 4 (CTLA-4) and cell death protein 1 pathway (PD-1), are rapidly being approved for multiple cancer types, including as first line therapy for PD-L1-expressing non-small-cell lung cancer. The combination of ACT and checkpoint blockade could substantially boost the efficacy of ACT. In this study, we generated a novel self-delivering small interfering RNA (siRNA) (sdRNA) that knocked down PD-1 expression on healthy donor T cells as well as patient-derived tumor-infiltrating lymphocytes (TIL). We have developed an alternative chemical modification of RNA backbone for improved stability and increased efficacy. Our results show that T cells treated with sdRNA specific for PD-1 had increased interferon γ (IFN-γ) secreting capacity and that this modality of gene expression interference could be utilized in our rapid expansion protocol for production of TIL for therapy. TIL expanded in the presence of PD-1-specific sdRNA performed with increased functionality against autologous tumor as compared to control TIL. This method of introducing RNAi into T cells to modify the expression of proteins could easily be adopted into any ACT protocol and will lead to the exploration of new combination therapies.

Skeletal changes following surgically assisted rapid maxillary expansion (SARME).

PURPOSE: Surgically assisted rapid maxillary expansion (SARME) is a common treatment technique in the correction of maxillary transverse deficiency. The purpose of this study was to evaluate nasal and palatal skeletal changes following SARME using Cone Beam Computed Tomography (CBCT) and posterior anterior (PA) cephalograms. MATERIALS AND METHODS: In this retrospective study, the radiographic images obtained from 14 patients with transverse maxillary deficiency before treatment and 6 months after SARME operations were evaluated. The changes in nasal bone width and palatal bone width were measured on CBCT. The changes in basal maxillary width, nasal cavity width and angular measurements were evaluated on PA cephalograms. RESULTS: Nasal floor width was measured at the levels of upper first premolar teeth and molar teeth which significantly increased following SARME (p=0.005 and 0.017 respectively). Palatal bone width between first premolar teeth and molar teeth also significantly increased (p=0.003 and 0.002 respectively). Basal maxillary width (p=0.026), nasal cavity width (p=0.024) and other angular measurements also significantly increased (p<0.05). CONCLUSION: Nasal and palatal skeletal transverse dimensions increased following SARME. Due to the enlargement of the nasal floor and nasal cavity, it is likely to improve air pass through the nose.

Simultaneous microencapsulation of hydrophilic and lipophilic bioactives in liposomes produced by an ecofriendly supercritical fluid process.

Organic solvent residues are always a concern with the liposomes produced by traditional techniques. Our objectives were to encapsulate hydrophilic and lipophilic compounds in liposomes using a newly designed supercritical fluid process coupled with vacuum-driven cargo loading. Supercritical carbon dioxide was chosen as the phospholipid-dissolving medium and an ecofriendly substitute for organic solvents. Liposomal microencapsulation was conducted via a 1000-µm expansion nozzle at 12.41MPa, 90°C, and aqueous cargo loading rate of 0.25ml/s. Vitamins C and E were selected as model hydrophilic and lipophilic compounds encapsulated in the integrated liposomes. The average vesicle size was 951.02nm with a zeta potential of -51.87mV. The encapsulation efficiency attained was 32.97% for vitamin C and 99.32% for vitamin E. Good emulsion stability was maintained during storage at 4°C for 20days. Simultaneous microencapsulation in the liposomes was successfully achieved with this supercritical fluid process.

Microencapsulation and characterization of liposomal vesicles using a supercritical fluid process coupled with vacuum-driven cargo loading.

A new technique of liposomal microencapsulation, consisting of supercritical fluid extraction followed by rapid expansion of the supercritical solution and vacuum-driven cargo loading, was successfully developed. It is a continuous flow-through process without usage of any toxic organic solvent. For use as a coating material, the solubility of soy phospholipids in supercritical carbon dioxide was first determined using a dynamic equilibrium system and the data was correlated with the Chrastil model with good agreement. Liposomes were made with D-(+)-glucose as a cargo and their properties were characterized as functions of expansion pressure, temperature, and cargo loading rates. The highest encapsulation efficiency attained was 31.7% at the middle expansion pressure of 12.41MPa, highest expansion temperature of 90°C, and lowest cargo loading rate of 0.25mL/s. The large unilamellar vesicles and multivesicular vesicles were observed to be a majority of the liposomes produced using this eco-friendly process.

Evaluation of the Potential Use of Laminar Extrudates on Stabilizing Micronized Coumarin Particles by Supercritical Fluids (RESS)-Study of Different RESS Processing Variables and Mode of Operation.

The study evaluates the ability of extrudates to deliver coumarin particles micronized by the rapid expansion of supercritical solutions (RESS). The RESS parameters were drug load (2-50 g), pressure (15-42 MPa) and temperature (40-60°C) in the extraction and pressure in the expansion (0.1-5 MPa) chambers in batch or continuous and CO2 flow rate in the continuous mode of operation. Particles were characterized for size (laser diffractometry, optical and electronic microscopies-19-61 µm), surface area (BET-0.282-0.423 m2/g), density (pycnometry-1.273-1.358 g/cm3) and yield (2-70%). Extrudates were characterized for the force of extrusion (4 kN), release of coumarin (100%/24 h) and mechanical properties (bending strength and stiffness increased, whereas elasticity decreased in storage) and X-ray diffractometry (micronized particles and extrudates have shown identical patterns) and calorimetry (DSC, enthalpies increased on storage). In the discontinuous mode of operation, increased loads in the extraction or increased pressure in the expansion chambers led to larger particles, whereas increased temperature and pressure in the extraction chamber led to smaller particles. In the continuous mode of operation, a decrease on the expansion pressure, load and CO2 flow rate led to increased yields. An increase on the flow rate led to a decrease on the particles' diameter, but an increase on coumarin load in the extraction chamber led to an increase in diameter. The study has identified the key parameters in RESS continuous and discontinuous modes of operation affecting the properties of the micronized coumarin particles and has proved the ability of extrudates with a laminar shape on delivering micronized particles.

Particle Formation and Product Formulation Using Supercritical Fluids.

Traditional methods for solids processing involve either high temperatures, necessary for melting or viscosity reduction, or hazardous organic solvents. Owing to the negative impact of the solvents on the environment, especially on living organisms, intensive research has focused on new, sustainable methods for the processing of these substances. Applying supercritical fluids for particle formation may produce powders and composites with special characteristics. Several processes for formation and design of solid particles using dense gases have been studied intensively. The unique thermodynamic and fluid-dynamic properties of supercritical fluids can be used also for impregnation of solid particles or for the formation of solid powderous emulsions and particle coating, e.g., for formation of solids with unique properties for use in different applications. We give an overview of the application of sub- and supercritical fluids as green processing media for particle formation processes and present recent advances and trends in development.

Phase I trial of adoptively transferred tumor-infiltrating lymphocyte immunotherapy following concurrent chemoradiotherapy in patients with locoregionally advanced nasopharyngeal carcinoma.

Adoptive cell therapy (ACT) for cancers using autologous tumor-infiltrating lymphocytes (TILs) can induce immune responses and antitumor activity in metastatic melanoma patients. Here, we aimed to assess the safety and antitumor activity of ACT using expanded TILs following concurrent chemoradiotherapy (CCRT) in patients with locoregionally advanced nasopharyngeal carcinoma (NPC). Twenty-three newly diagnosed, locoregionally advanced NPC patients were enrolled, of whom 20 received a single-dose of TIL infusion following CCRT. All treated patients were assessed for toxicity, survival and clinical and immunologic responses. Correlations between immunological responses and treatment effectiveness were further studied. Only mild adverse events (AEs), including Grade 3 neutropenia (1/23, 5%) consistent with immune-related causes, were observed. Nineteen of 20 patients exhibited an objective antitumor response, and 18 patients displayed disease-free survival longer than 12 mo after ACT. A measurable plasma Epstein-Barr virus (EBV) load was detected in 14 patients at diagnosis, but a measurable EBV load was not found in patients after one week of ACT, and the plasma EBV load remained undetectable in 17 patients at 6 mo after ACT. Expansion and persistence of T cells specific for EBV antigens in peripheral blood following TIL therapy were observed in 13 patients. The apparent positive correlation between tumor regression and the expansion of T cells specific for EBV was further investigated in four patients. This study shows that NPC patients can tolerate ACT with TILs following CCRT and that this treatment results in sustained antitumor activity and anti-EBV immune responses. A larger phase II trial is in progress.

Simplified protocol for clinical-grade tumor-infiltrating lymphocyte manufacturing with use of the Wave bioreactor.

BACKGROUND AIMS: The high level of complexity of current Good Manufacturing Practice-compliant methods of manufacturing hampers rapid and broad application of treatment with tumor-infiltrating lymphocytes (TILs). METHODS: To ensure higher applicability of TIL production to laboratory routine, a practical and simple protocol of TIL manufacturing with the use of a closed-system bioreactor was developed and implemented at our institution. RESULTS: This protocol enabled significant work load reduction during the most labor-intense step of TIL expansion, and allowed generation of high-quality TIL products, which mediated clinical regression in patients with metastatic melanoma. CONCLUSIONS: Implementation of simplified methods of TIL expansion will speed up dissemination of TIL methods worldwide and will increase patient access to this highly effective treatment.

Longitudinal quantitative evaluation of the mid-palatal suture after rapid expansion using in vivo micro-CT.

New bone formation is known to occur between the opened palatal bones after rapid mid-palatal expansion (RME), although the time-dependent changes in the mid-palatal suture after RME have not been fully examined. Thus, we investigated time-dependent morphological changes in the mid-palatal suture using in vivo micro-computed tomography (mCT) and the expression of bone morphogenetic factors. RME was performed by inserting a 1.5-mm-thick circular metal ring between the maxillary incisors of rats, and morphological changes in the mid-palatal suture were investigated using in vivo mCT imaging after RME. Bone morphogenetic protein 2 (BMP-2) and insulin-like growth factor-I (IGF-I) expression in the suture were also examined using reverse-transcription polymerase chain reaction and immunohistochemistry. The bone volume of the mid-palatal suture decreased after RME to a minimum of -0.34mm(3) on day 12, then increased with bone formation over time and reached -0.13mm(3) on day 24. Significant increases in BMP-2 and IGF-I mRNA expression after RME were found on day 3 compared with day 0. By immunohistochemistry, BMP-2 and IGF-I were detected in osteoblasts on days 5 and 7, in endothelial cells of blood vessels, and fibroblasts on day 7. Expansion of the mid-palatal suture continues for 12 days after a single RME, and restoration requires more than 30 days. Additionally, BMP-2 and IGF-I may play important roles in the restoration process.

Maxillary protraction and multiple loop edgewise arch wire technology in treatment of early mixed dentition skeletal class Ⅲ malocclusion / 中华医学美学美容杂志

Objective To evaluate the orthodontic effect of surgery on mixed dentition period class Ⅲ malocclusion and to investigate the effective therapeutic method of the skeletal class Ⅲ malocclusion.Methods A total of 20 patients aged 8.5 to 9.1 years with early mixed dentition class Ⅲ skeletal malocclusion.The treatment was divided into two phases.During the first phase,the entire stuffy group was treated with a bonded maxillary expansion and protraction.During the second phase,the entire study group received multiple loop edgewise arch wire (MEAW) treatment until an ideal occlusion relationship achieved.Comparison of the pre-and post-orthodontic treatment cephalometric radiographs had been done after maxillary protraction and MEAW were applied for patients.Results In the 20 patients after orthodontic treatment,Class Ⅲ skeletal facial profile was improved,the retraction of anterior teeth was obvious,and there were significant changes in dentition; the profile of soft tissue and satisfactory occlusion relationship were significantly improved,as compared with the changes between pre-treatment and post-treatment.Conclusions Skeletal Ⅲ malocclusion in early mixed dentition can be successfully treated.The results can be satisfied with improved class Ⅲ skeletal and molar relationship,high-angle and profile of soft tissue.