Sterile Conditions Research Articles

Lipidomic scanning of self-lipids identifies headless antigens for natural killer T cells

To broadly measure the spectrum of cellular self-antigens for natural killer T cells (NKT), we developed a sensitive lipidomics system to analyze lipids trapped between CD1d and NKT T cell receptors (TCRs). We captured diverse antigen complexes formed in cells from natural endogenous lipids, with or without inducing endoplasmic reticulum (ER) stress. After separating protein complexes with no, low, or high CD1d-TCR interaction, we eluted lipids to establish the spectrum of self-lipids that facilitate this interaction. Although this unbiased approach identified fifteen molecules, they clustered into only two related groups: previously known phospholipid antigens and unexpected neutral lipid antigens. Mass spectrometry studies identified the neutral lipids as ceramides, deoxyceramides, and diacylglycerols, which can be considered headless lipids because they lack polar headgroups that usually form the TCR epitope. The crystal structure of the TCR-ceramide-CD1d complex showed how the missing headgroup allowed the TCR to predominantly contact CD1d, supporting a model of CD1d autoreactivity. Ceramide and related headless antigens mediated physiological TCR binding affinity, weak NKT cell responses, and tetramer binding to polyclonal human and mouse NKT cells. Ceramide and sphingomyelin are oppositely regulated components of the "sphingomyelin cycle" that are altered during apoptosis, transformation, and ER stress. Thus, the unique molecular link of ceramide to NKT cell response, along with the recent identification of sphingomyelin blockers of NKT cell activation, provide two mutually reinforcing links for NKT cell response to sterile cellular stress conditions.

Collagen Membrane as Artificial Dura Substitute: A Comprehensive In Vivo Study of Efficiency and Substitution Compared to Durepair

The dura mater is a barrier between the brain and the surrounding environment. Injuries to the dura can lead to serious complications, therefore, ensuring a hermetic closure of the dura is a primary task for a neurosurgeon. The aim of the study is to compare the effectiveness of applying the newly developed ViscollDURA collagen membrane (VDCM), with the commercially available Durepair (xenogeneic collagen) in animal model. A dural tear model was utilized in rats with membrane implantation using an application method. The sample size consisted of 24 rats. Group I underwent VDCM implantation, while Group II underwent Durepair implantation. Results were evaluated at 30, 60, and 90days. The study was assessed using magnetic resonance imaging, histology, electron scanning microscopy, and immunohistochemistry. The obtained results underwent statistical analysis. In the clinical presentation, there were no difference between groups. Histologically, Group 1 showed comparable results to Group 2. The integration process of the membrane statistically differed between the groups. In Group 1, neovascularization and tissue replacement showed better results than in Group 2. Magnetic resonance imaging differences were observed at later stages, with group 2 showing adhesion and brain deformation in the implantation area. Both membranes showed safety and compatibility. The collagen membrane produced under sterile conditions demonstrated better regeneration with minimal inflammatory reaction. The study suggests that VDCM exhibits biocompatibility comparable to Durepair, providing prospects for potential applications in neurosurgery.

Machine learning-based bee recognition and tracking for advancing insect behavior research

The study of insect behavior, particularly that of honey bees, has a broad scope and significance. Tracking bee flying patterns grants much helpful information about bee behavior. However, tracking a small yet fast-moving object, such as a bee, is difficult. Hence, we present artificial intelligence, machine-learning-based bee recognition, and tracking systems to assist the researcher in studying the bee’s behavior. To develop a machine learning system, a labeled database is required for model training. To address this, we implemented an automated system for analyzing and labeling bee videos. This labeled database served as the foundation for two distinct bee-tracking solutions. The first solution (planar bee tracking system) tracked individual bees in closed mazes using a neural network. The second solution (spatial bee tracking system) utilized a neural network and a tracking algorithm to identify and track flying bees in open environments. Both systems tackle the challenge of tracking small-bodied creatures with rapid and diverse movement patterns. Although we applied these systems to entomological cognition research in this paper, their relevance extends to general insect research and developing tracking solutions for small organisms with swift movements. We present the complete architecture and detailed methodologies to facilitate the utilization of these models in future research endeavors. Our approach is a simple and inexpensive method that contributes to the growing number of image-analysis tools used for tracking animal movement, with future potential applications under less sterile field conditions. The tools presented in this paper could assist the study of movement ecology, specifically in insects, by providing accurate movement specifications. Following the movement of pollinators or natural enemies, for example, greatly contributes to the study of pollination or biological control, respectively, in natural and agro-ecosystems.

Intravenous delivery of furosemide using lipid-based versus polymer-based nanocapsules: in vitro and in vivo studies

Objectives Furosemide (FSM), a potent loop diuretic, is used to treat edema due to hypertension, congestive heart failure, and liver and renal failures. FSM applications are limited by its low bioavailability. Our aim is to use different nanoencapsulation strategies to control the release of FSM and enhance its pharmacokinetic properties. Methods Two types of FSM-loaded nanocapsules, namely FSM-loaded lipid nanocapsules (LNCs) and polymeric nanocapsules (PNCs), were developed, physicochemically characterized, and subjected to pharmacokinetic and pharmacodynamic studies. Lipid nanocapsules were prepared by the simple phase inversion method using LabrafacTM lipid, while the polymeric nanocapsules were prepared by nanoprecipitation method using polycaprolactone polymer. Results Transmission electron microscopy ascertains spherical structures, corroborating the nanometric diameter of both types of nanocapsules. The particle size of the optimized FSM-loaded LNCs and FSM-loaded PNCs was 32.19 ± 0.72 nm and 230.7 ± 5.13 nm, respectively. The percent entrapment efficiency was 63.56 ± 1.40% of FSM for the optimized PNCs. The in vitro release study indicated prolonged drug release compared to drug solutions. The two loaded nanocapsules systems succeeded in enhancing the pharmacokinetic parameters in comparison to the marketed FSM solution with superior diuretic activity (p < 0.05). The results of the stability study and the terminal sterilization by autoclave indicated the superiority of LNCs over PNCs in maintaining the physical parameters under storage conditions and the drastic conditions of sterilization. Conclusions LNCs and PNCs are considered promising nanosysems for improving the diuretic effect of FSM.

An in-vitro model for bacteria-related catheter encrustations.

About 50% of individuals with long-term indwelling catheters are affected by catheter encrustations and bladder stone formation. Therefore, prophylaxis of catheter encrustations is important. Currently, however, neither an established prophylaxis nor a standardized in-vitro model to test different measures exist. We have therefore developed and qualitatively evaluated an in-vitro model of catheter encrustation. Size 14 French suprapubic catheters were incubated under sterile conditions at 37 degrees Celsius in five different media: (1) sterile artificial urine (n = 16), (2) artificial urine with E. coli (n = 8), (3) with Pseudomonas aeruginosa (n = 8), (4) with Proteus mirabilis (n = 8), and (5) with a mix of these three strains (n = 8). Catheter balloons were inflated either a glycerine or a bactericidal solution. After 6 weeks, the catheters were removed from the solution, dried, and weighed, and a photometric determination of the retrieved encrustations was performed. Most frequently and pronounced encrustations were detected in the Pseudomonas group. The median weight of these encrustations (50% struvite and brushite) was 84.4mg (47.7mg / 127.3mg). Even on catheters stored in sterile urine, encrustations (69.2% struvite) were found. Bacterial growth was not affected by the medium used for catheter blockage. Although in-vitro models appear to be limited because they lack "the human factor", they are valuable for systematically assessing physico-chemical factors affecting encrustations. Therefore, our model, being reliable and cost-effective, may foster further research despite its limitations.

A Bioreactor for Celullarised Membrane Culture and Delivery under Sterile Conditions.

A novel, user-friendly bioreactor for the cultivation of cellularised membranes for tissue engineering has been successfully designed, manufactured, and validated. This bioreactor features a culture vessel and a cover, the latter equipped with one or more sidewalls to ensure airtightness in two distinct zones, thereby maintaining sterile conditions. The cover, designed to integrate seamlessly with the culture vessel, includes several ports compatible with commercial connectors. This design allows the introduction of cells and culture medium without requiring the opening of the cover, thus preserving sterility. Additionally, the cover is equipped with flanges that effectively press the membrane against the bottom surface of the culture vessel, preventing it from shrinking or shifting. This ensures that cells can properly adhere to the membrane and proliferate. Manufactured under Good Manufacturing Practice (GMP) conditions, the bioreactor supports cultivation in optimal aseptic environments, thereby preventing external contamination. This feature is critical for the safe transportation of cultivated tissue to clinical settings. Validation tests have confirmed the bioreactor's excellent performance, endorsing its suitability for intended applications in tissue engineering.

Edema and Trismus Assessment After Applying Kinesiotape With the Web Strip Technique Compared With Cryotherapy: A Randomized Clinical Trial.

Extraction of impacted third molars is one of the most common oral surgeries performed in the dental clinic, which is often accompanied by many complications such as edema and trismus. Many methods have been used to alleviate these complications, such as drugs or physical therapy. Kinesiotape (KT) has recently spread as a popular physical method for eliminating complications after surgical extraction of lower third molars after its long-term use in sports medicine and injuries of the musculoskeletal system. The current study aimed to study the effect of using KT (Kinesio® Holding Corporation, Albuquerque, NM, USA) on both edema and trismus after impacted third molar extraction. This study was designed as a randomized controlled clinical trial using the split-mouth technique and included 25 patients with radiographically symmetrical lower third molars from patients. All surgical extractions were performed by a single surgeon under sterile conditions according to the standard surgical protocol after that one group applied KT and the other group applied cryotherapy. Edema and trismus were measured in the first five days. Data was collected and analyzed by SPSS software (IBM Corp., Armonk, NY, USA). The mean change in the total linear facial measurements in the experimental group (Kinesio Group) was 12.32 mm three days after surgery, and then this value decreased to 6.80 mm, while the average increase in the control group (Cryo Group) was 17.00 mm after three days, then the value decreased to 9.68 mm five days after surgery. Regarding the changes in the amount of maximum mouth opening after surgery, the results were similar between the Kinesio Group and the Cryo Group, as there were no significant differences between the two groups (P<0.05). This study concluded that KT was superior to cryotherapy when studying edema. The current study also concluded that the mouth opening was similar between the two study groups.

가열살균조건에 따른 양파 착즙액 첨가 한식간장의 저장 중 품질 특성 변화

The objective of this study was to integrate onion juice into traditional Korean soy sauce and to evaluate the sterilization conditions necessary for commercialization. As the sterilization temperature increased, pH decreased and titratable acidity increased. A reduction in °Brix was observed post-sterilization, from 37.37 °Brix in untreated soy sauce to 36.67-36.77 °Brix. In terms of chromaticity, the L value declined over the storage period while the a and b values increased. The quercetin component exhibited light sensitivity, with no significant difference observed in the sample on the 60th day. Neither Bacillus cereus nor Clostridium perfringens were detected, and the total bacterial count was comparable to that of commercially available soy sauce. This indicates that samples sterilized at 85°C for 30 minutes fall within a safe quality range. Sensory evaluation revealed that samples sterilized at 85°C for 30 minutes exhibited consistently high ratings.

가열살균조건에 따른 양파 착즙액 첨가 한식간장의 저장 중 품질 특성 변화

The objective of this study was to integrate onion juice into traditional Korean soy sauce and to evaluate the sterilization conditions necessary for commercialization. As the sterilization temperature increased, pH decreased and titratable acidity increased. A reduction in °Brix was observed post-sterilization, from 37.37 °Brix in untreated soy sauce to 36.67-36.77 °Brix. In terms of chromaticity, the L value declined over the storage period while the a and b values increased. The quercetin component exhibited light sensitivity, with no significant difference observed in the sample on the 60th day. Neither Bacillus cereus nor Clostridium perfringens were detected, and the total bacterial count was comparable to that of commercially available soy sauce. This indicates that samples sterilized at 85°C for 30 minutes fall within a safe quality range. Sensory evaluation revealed that samples sterilized at 85°C for 30 minutes exhibited consistently high ratings.

Sodium Tanshinone IIA Sulfonate Protects Primary Cardiomyocytes Against Radiation-Induced Myocardial Injury via the p38 Pathway.

Sodium tanshinone IIA sulfonate (STS), which is extracted from a Chinese medicinal herb, possesses many pharmacologic functions, such as coronary dilation, anti-inflammatory properties, and antiapoptotic and antioxidant effects. It remains unknown whether STS can protect cardiomyocytes injured after radiation therapy. An in vitro Sprague-Dawley (SD) rat neonatal cardiomyocyte system was established. Primary cardiomyocytes (PCMs) from neonatal SD rats were isolated under sterile conditions. PCM cells were divided into a control group (0 Gy/hour) and 5 experimental radiation therapy groups (0.25 Gy/hour, 0.5 Gy/hour, 1 Gy/hour, 2 Gy/hour, and 4 Gy/hour). Cell viability, the content of malondialdehyde (MDA), the lactate dehydrogenase (LDH) leakage rate, and superoxide dismutase (SOD) and glutathione (GSH) activities were recorded separately in each group after 7 days of culture. Western blot was used to detect the levels of p38, caspase-3 protein, and X protein (BAX) associated with B-cell lymphoma 2 (Bcl-2) in PCMs. X-rays inhibited cell growth, decreased cell viability, and induced an oxidative stress response in PCMs. STS and SB203580 (the inhibitor of P38 mitogen-activated protein kinase pathway) alleviated X-ray-induced damage to PCMs. An enzyme-linked immunosorbent assay showed that X-rays increased the cTnT level. STS and SB203580 ameliorated the X-ray-induced increase in cTnT leakage. X-rays enhanced the expression of p38/p-p38 and caspase-3 while reducing the expression of Bcl-2/BAX in PCMs, as demonstrated by western blotting. STS and SB203580 mitigated the changes in protein expression triggered by X-ray radiation. In conclusions, STS was shown to exert significant cardioprotective, anti-inflammatory, and antioxidant effects in PCMs by inhibiting the p38 mitogen-activated protein kinase pathway.

In vitro corrosion-fatigue behaviour of rare-earth containing magnesium WE43 in sterile complex cell culture medium

Rare-earth containing magnesium alloys are promising biomedical materials for a new generation of biodegradable orthopaedic implant systems due to their excellent biocompatibility, mechanical and biodegradation properties. However, chemo-mechanical interactions in aggressive physiological corrosion environments result in rapid degradation and early loss of mechanical integrity, limiting its broader application for orthopaedic implants. To date, only few studies have assessed the corrosion-fatigue behaviour of medical-grade magnesium alloys in an organic physiological corrosion environment, especially under sterile test conditions. In the present work, the corrosion-fatigue behaviour of fine-grained medical-grade magnesium alloy WE43MEO was systematically analysed under in vitro conditions using an organic physiological fluid DMEM. The experimental results showed that the fatigue strength of the alloy is nearly unaffected by a 1-day precorrosion, while a 7-day precorrosion resulted in a significant deterioration of mechanical integrity. In corrosion-fatigue experiments, the fatigue life was considerably reduced by interactions between corrosion and fatigue damages. The SEM analysis revealed that the mixed mode of intergranular and transgranular fracture in the crack propagation zone transits to intergranular cracking dominant mode under the corrosion-fatigue conditions due to hydrogen embrittlement.

Innovative plasma treatment of orange juice to improve bioactive concentration: The effects of various parameters using response surface analysis.

Orange juice is a highly nutritious beverage. Traditional pasteurization methods cause nutrient loss and taste changes. Plasma treatment (PT) is an emerging method with a high sterilization rate. This study investigated the effects of corona discharge plasma on the sterilization of orange juice by changes in color difference, total phenol content, and pH value. Single-factor experiments revealed that higher voltage (40kV) and longer sterilization time (25min) had better sterilization effects. Response surface analysis indicated that frequency had the greatest impact on sterilization rates, and the optimal sterilization conditions were a voltage of 44.75kV, a frequency of 9.46kHz, and a sterilization time of 25min. Under these conditions, the sterilization rate reached 97.9%, meeting the national standard of 104 colony-forming units/mL (GB7101-2022). Compared to untreated juices, the color difference value was 16.32, the pH value decreased by 0.12, and the total phenol content increased by 0.669mg/mL. However, the evaporation of water plays an important role in increasing the total phenol co. Moreover, the comparative analysis showed that PT was comparable to pasteurization in terms of sterilization effects, flavor preservation, and the concentration of bioactive components. This study provides a theoretical basis for industrial applications of PT.

Advanced Sperm Selection Techniques for Assisted Reproduction.

Male infertility accounts for approximately 40% of infertility cases. There are many causes of male infertility, including environmental factors, age, lifestyle, infections, varicocele, and cancerous pathologies. Severe oligozoospermia, cryptozoospermia, and azoospermia (obstructive and non-obstructive) are identified as severe male factor infertility, once considered conditions of sterility. Today, in vitro fertilization (IVF) techniques are the only treatment strategy in cases of male factor infertility for which new methodologies have been developed in the manipulation of spermatozoa to achieve fertilization and increase success rates. This review is an update of in vitro manipulation techniques, in particular sperm selection, emphasizing clinical case-specific methodology. The success of an IVF process is related to infertility diagnosis, appropriate choice of treatment, and effective sperm preparation and selection. In fact, selecting the best spermatozoa to guarantee an optimal paternal heritage means increasing the blastulation, implantation, ongoing pregnancy and live birth rates, resulting in the greater success of IVF techniques.

P-279 Significant increased isolation of escherichia coli in Iranian women with endometriosis: a case control – study

Abstract Study question Considering the role of bacterial infection in causing endometriosis, we decided to compare the cultures of endometriosis-affected women a in this study Summary answer Considering the possible role of E. coli in the progression and/or recurrence of endometriosis and also infertility What is known already (5). Due to the finding of endotoxin of E. coli species in the peritoneal cavity and menstrual fluid of endometriosis patients, infection with this bacterium has recently been proposed as an effective factor in the occurrence and growth of endometriosis lesions. Study design, size, duration This prospective case-control study was carried out in the Women’s Department of Shiraz University of Medical Sciences in collaboration with the Bacteriology and Virology Department between 60 participants. Participants/materials, setting, methods 30 women with endometriosis in stages 3 and 4 of the disease, and 30 women without endometriosis who were candidates for surgery due to benign uterine diseases, underwent endometrial biopsy with Novak Kort in sterile conditions before starting their operation, and the results of their endometrial culture were analyzed and compared in terms of the presence of Enterobacter spp., Klebsiella spp., and E. coli species. Main results and the role of chance Based on the comparison between the two groups, only E. coli species had a significant association with the presence of endometriosis (P = 0.001). There was no significant relationship between the location of endometriosis in the pelvic cavity and culture results. Parity among the E. coli negative group was significantly higher compared to the E. coli positive group (P &amp;lt; 0.001). Limitations, reasons for caution Lack of sample trail for repeat testing and antibiotic susceptibility testing, low taxonomical resolution, and costly laboratory setup and maintenance are other limitations of this technology Wider implications of the findings we suggest that in women who have endometriosis and recurrent IVF failure and also in cases of endometriosis recurrence after surgical treatment, treatment of couples with appropriate antibiotics and repeated culture along with subsequent antibiotic susceptibility testing until their culture becomes negative could be beneficial. Trial registration number IR.SUMS.MED.REC.1400.097

Cord Blood Hemoglobin Levels in Full Term Neonates of Mothers with Iron Deficiency Anemia

Abstract Background Iron is a critical micronutrient for tissue oxygenation, cellular metabolism, energy generation, and the metabolism of toxins. Iron transfer from mother to fetus is a regulated process involving iron status in the maternal circulation, its transport across the placenta and subsequent transfer into the fetal circulation. Objective To evaluate cord blood Hemoglobin (Hb) levels in neonates of mothers with iron deficiency anemia (IDA). Patients and Methods This prospective case-control study was conducted at Ain Shams University Hospitals. It included 60 full terms newborn who were born at Obstetrics and Gynecology Hospital. The study population included full term neonates delivered at ≥ completed 37 weeks of gestation which was divided into two groups according to maternal risk factor: Case: full term neonates with maternal risk factor. Control: full term neonates without maternal risk factor. Thorough clinical examination and investigations were performed including cord blood samples collected for measurement of the level of complete blood count (CBC; (Hb, MCV and MCH). For the mothers, venous blood samples were withdrawn with standard precautions and strict sterile conditions for CBC, transferrin saturation and serum ferritin. Results In comparison between cases and controls as regard RBCS parameter, there was no significant difference between cases and controls regarding neonatal hemoglobin( 14.93±1.14 vs 15.44 ±1.35(g/dL), p = 0.12) and MCH (32.51±2.9 vs33.57 ±1.9 (pg), p = 0.10), but there was significantly lower MCV in cases compared to controls (104.07± 7.36 vs 109.60±4.93 (fL), p = 0.001). There was no significant difference between cases and controls as regard maternal clinical characteristics including (age, weight, height, maternal diseases, gravidity, consanguinity and mode of delivery, p &amp;gt; 0.05 for all). Also, there was no significant difference between cases and controls regarding neonatal weight, Z score for weight, length (cm), head circumference (cm) and gestational age by Ballord Score and Apgar Score in 1-minute and 5-minute score p &amp;gt; 0.05 for all). Conclusion There was no relation between the maternal Hb deficient levels and neonatal Hb. The first sign of iron deficiency in neonates reflected in lower MCV in cases, but not reached to decrease Hb levels in neonates.

Immunological foundations of neurological diseases

Neurodegenerative diseases represent an increasing challenge in ageing societies, as only limited treatment options are currently available. New research methods and interdisciplinary interaction of different disciplines have changed the way neurological disorders are viewed and paved the way for the comparatively new field of neuroimmunology, which was established in the early 1980s. Starting from neurological autoimmune diseases, such as multiple sclerosis, knowledge about the involvement of immunological processes in other contexts, such as stroke or traumatic brain injury, has been significantly expanded in recent years. This review article provides an overview of the role of the immune system and the resulting potential for novel treatment approaches. The immune system plays acentral role in fighting infections but is also able to react to the body's own signals under sterile conditions and cause inflammation and subsequent adaptive immune responses through the release of immune mediators and the recruitment and differentiation of certain immune cell types. This can be beneficial in initiating healing processes; however, chronic inflammatory conditions usually have destructive consequences for the tissue and the organism and must be interrupted. It is now known that different cells of the immune system play an important role in neurological diseases. Regulatory mechanisms, which are mediated by regulatory Tcells or Th2 cells, are usually associated with agood prognosis, whereas inflammatory processes and polarization towards Th1 or Th17 have adestructive character. Novel immunomodulators, which are also increasingly being used in cancer treatment, can now be used in atissue-specific manner and therefore offer great potential for use in neurological diseases.

Radioactivity and GMO-Free Sterile Insect Technology for the Sustainable Control of the Invasive Pest Drosophila suzukii.

Drosophila suzukii (D. suzukii), commonly known as the spotted wing drosophila, is a highly invasive crop pest that is difficult to control using chemical insecticides. To address the urgent need for alternative and more sustainable control strategies, the sterile insect technique (SIT) is improved, which involves the release of sterilized male insects to mate with fertile conspecifics, thereby reducing the size of the pest population in the subsequent generation. The three critical aspects that influence the success of SIT programs in D. suzukii are addressed. First, an accurate and nondestructive method is established to determine the sex of individual insects based on the differential weight of male and female pupae. Second, conditions for X-ray sterilization are systematically tested and an optimal dose (90kV/40Gy) is identified that ensures the efficient production of sterile D. suzukii for release. Finally, the inherent thermosensitivity of D. suzukii males is exploited to develop a temperature-based sterilization technique, offering an alternative or additional SIT method for this pest. These advances will contribute to the development of a comprehensive and effective strategy for the management of D. suzukii populations, reducing their impact on agriculture and helping to safeguard crop yields.

Highly stable, fluorescent, artificial lipoprotein nanoparticles.

Here, we report a novel kind of protein nanoparticles of 11 nm in size, which have a central protein core surrounded by two layers of lipid. One layer of the lipid was covalently attached to the protein, while the other layer has been physically assembled around the protein core. Particle synthesis is highly modular, while both the size and charge of the protein nanoparticles are controlled in a predictable manner. Circular dichroism studies of the conjugate showed that the protein secondary structure is retained, while biophysical characterizations indicated the particle purity, size, and charge. The conjugate had a high thermal stability to steam sterilization conditions at 121°C (17 psi). After labeling the protein core with few different fluorescent dyes, they were strongly fluorescent with the corresponding colors independent of their size, unlike quantum dots. They are readily digested by proteases, and these water-soluble, non-toxic, highly stable, biocompatible, and biodegradable conjugates are suitable for cell imaging and drug delivery applications.

Effect and mechanisms of thermal sterilization methods on the in vitro phenolic bioaccessibility of rose tea with milk

Rose polyphenols, key functional components in roses, require adequate bioaccessibility for their health benefits, subject to influence by food components and processing. Investigating the impact of various thermal sterilization methods on the bioaccessibility of rose polyphenols in rose tea with milk and the underlying mechanisms, our findings indicated a significant increase in bioaccessibility following treatment at 85 °C/30 min. Conversely, 121 °C/15 min treatment decreased bioaccessibility. Examining the interaction between β-casein in milk and rose polyphenols under different sterilization conditions, SEM and particle size analysis revealed binding, with fluorescence spectroscopy indicating non-covalent bonds. Binding forces followed the order 121 °C > 85 °C > 25 °C. Notably, at 85 °C, non-covalent binding improved polyphenol bioaccessibility, while the intensified binding at 121 °C decreased it. SDS-PAGE and amino acid analysis confirmed no covalent bond. This study establishes a theoretical basis for selecting thermal sterilization temperatures for milk-flower combinations, considering polyphenol bioaccessibility.

Development of Focused Ultrasound-Assisted Nanoplexes for RNA Delivery.

RNA-based therapeutics, including siRNA, have obtained recognition in recent years due to their potential to treat various chronic and rare diseases. However, there are still limitations to lipid-based drug delivery systems in the clinical use of RNA therapeutics due to the need for optimization in the design and the preparation process. In this study, we propose adaptive focused ultrasound (AFU) as a drug loading technique to protect RNA from degradation by encapsulating small RNA in nanoliposomes, which we term nanoplexes. The AFU method is non-invasive and isothermal, as nanoplexes are produced without direct contact with any external materials while maintaining precise temperature control according to the desired settings. The controllability of sample treatments can be effectively modulated, allowing for a wide range of ultrasound intensities to be applied. Importantly, the absence of co-solvents in the process eliminates the need for additional substances, thereby minimizing the potential for cross-contaminations. Since AFU is a non-invasive method, the entire process can be conducted under sterile conditions. A minimal volume (300 μL) is required for this process, and the treatment is speedy (10 min in this study). Our in vitro experiments with silencer CD44 siRNA, which performs as a model therapeutic drug in different mammalian cell lines, showed encouraging results (knockdown > 80%). To quantify gene silencing efficacy, we employed quantitative polymerase chain reaction (qPCR). Additionally, cryo-electron microscopy (cryo-EM) and atomic force microscopy (AFM) techniques were employed to capture images of nanoplexes. These images revealed the presence of individual nanoparticles measuring approximately 100-200 nm in contrast with the random distribution of clustered complexes observed in ultrasound-untreated samples of liposome nanoparticles and siRNA. AFU holds great potential as a standardized liposome processing and loading method because its process is fast, sterile, and does not require additional solvents.