Initial Release Research Articles

Large-scale releases and establishment of wMel Wolbachia in Aedes aegypti mosquitoes throughout the Cities of Bello, Medellín and Itagüí, Colombia.

The wMel strain of Wolbachia has been successfully introduced into Aedes aegypti mosquitoes and has been shown to reduce the transmission of dengue and other Aedes-borne viruses. Here we report the entomological results from phased, large-scale releases of Wolbachia infected Ae. aegypti mosquitoes throughout three contiguous cities located in the Aburrá Valley, Colombia. Local wMel Wolbachia-infected Ae. aegypti mosquitoes were generated and then released in an initial release pilot area in 2015-2016, which resulted in the establishment of Wolbachia in the local mosquito populations. Subsequent large-scale releases, mainly involving vehicle-based releases of adult mosquitoes along publicly accessible roads and streets, were undertaken across 29 comunas throughout Bello, Medellín and Itagüí Colombia between 2017-2022. In 9 comunas these were supplemented by egg releases that were undertaken by staff or community members. By the most recent monitoring, Wolbachia was found to be stable and established at consistent levels in local mosquito populations (>60% prevalence) in the majority (67%) of areas. These results, from the largest contiguous releases of wMel Wolbachia mosquitoes to date, highlight the operational feasibility of implementing the method in large urban settings. Based on results from previous studies, we expect that Wolbachia establishment will be sustained long term. Ongoing monitoring will confirm Wolbachia persistence in local mosquito populations and track its establishment in the remaining areas.

Ca2+ regulation of glutamate release from inner hair cells of hearing mice

In our hearing organ, sound is encoded at ribbon synapses formed by inner hair cells (IHCs) and spiral ganglion neurons (SGNs). How the underlying synaptic vesicle (SV) release is controlled by Ca2+ in IHCs of hearing animals remained to be investigated. Here, we performed patch-clamp SGN recordings of the initial rate of release evoked by brief IHC Ca2+-influx in an ex vivo cochlear preparation from hearing mice. We aimed to closely mimic physiological conditions by perforated-patch recordings from IHCs kept at the physiological resting potential and at body temperature. We found release to relate supralinearly to Ca2+-influx (power, m: 4.3) when manipulating the [Ca2+] available for SV release by Zn2+-flicker-blocking of the single Ca2+-channel current. In contrast, a near linear Ca2+ dependence (m: 1.2 to 1.5) was observed when varying the number of open Ca2+-channels during deactivating Ca2+-currents and by dihydropyridine channel-inhibition. Concurrent changes of number and current of open Ca2+-channels over the range of physiological depolarizations revealed m: 1.8. These findings indicate that SV release requires ~4 Ca2+-ions to bind to their Ca2+-sensor of fusion. We interpret the near linear Ca2+-dependence of release during manipulations that change the number of open Ca2+-channels to reflect control of SV release by the high [Ca2+] in the Ca2+-nanodomain of one or few nearby Ca2+-channels. We propose that a combination of Ca2+ nanodomain control and supralinear intrinsic Ca2+-dependence of fusion optimally links SV release to the timing and amplitude of the IHC receptor potential and separates it from other IHC Ca2+-signals unrelated to afferent synaptic transmission.

Experimental investigation of gas diffusion kinetics and pore-structure characteristics during coalbed methane desorption within a coal seam

The desorption-diffusion properties of coalbed methane (CBM) are not only the theoretical basis for investigating gas outburst hazards in a coal mine, but also impose a non-negligible impact on CBM recovery performance. In this paper, the pore structure of coal with five different metamorphism degrees was qualitatively and quantitatively characterized with the scanning electron microscopy (SEM) and low-temperature nitrogen gas adsorption (LTNGA) measurements. The isothermal methane ad/desorption experiments were performed with four initial gas desorption pressure scales. Subsequently, the matching degree between eight empirical correlations (ECs) and the measured methane desorption data was determined, and the methane diffusion coefficient (Dfg) was numerically calculated combining with an FGDG (free gas density gradient) diffusion model. Importantly, we evaluated and discussed the sensitivity of pore structure parameters, initial gas desorption pressure, and volatile fraction to Dfg. The results show that pore density distribution differs among coal samples and the pore structure, especially the micropores, is developed better in high-rank coals than in low- and medium-rank ones. Initial gas release velocity increases with an increase in coal rank and decreases negatively with the increasing particle size in an exponential manner. Among the eight ECs, EC-VIII can be applied to more accurately describe gas desorption process during the whole time scale, and its correlation coefficient with the measured data is larger than 0.9900. Dfg increases with an increase in pore volume and specific surface area (SSA), and it is negatively correlated with the average pore diameter. Additionally, Dfg exhibits an asymmetric U-shaped pattern with an increase in the volatile fraction, which is mainly related to the switch between the dominant roles of mesopores and micropores during coal metamorphism. At a lower volatile fraction, the diffusion coefficient is more sensitive to micropores, while, at a higher volatile fraction, mesopores contribute more to the diffusion coefficient.

Extension of the glmm.hp package to zero-inflated generalized linear mixed models and multiple regression

Abstract glmm.hp is an R package designed to evaluate the relative importance of collinear predictors within generalized linear mixed models (GLMMs). Since its initial release in January 2022, it has been rapidly gained recognition and popularity among ecologists. However, the previous glmm.hp package was limited to work GLMMs derived exclusively from the lme4 and nlme packages. The latest glmm.hp package has extended its functions. It has integrated results obtained from the glmmTMB package, thus enabling it to handle zero-inflated generalized linear mixed models (ZIGLMMs) effectively. Furthermore, it has introduced the new functionalities of commonality analysis and hierarchical partitioning for multiple linear regression models by considering both unadjusted R2 and adjusted R2. This paper will serve as a demonstration for the applications of these new functionalities, making them more accessible to users.

LDPE/PCL nanofibers embedded chlorhexidine diacetate for potential antimicrobial applications

In this study, we described the process of developing antimicrobial nanofibers using a solution-based electrospinning technique. The nanofibers were composed of a 40:60 blend of low-density polyethylene (LDPE) and polycaprolactone (PCL), loaded with chlorhexidine diacetate (CHD).The diameter of the nanofibers was measured for LDPCL (235 nm), but it changed to 148, 194, and 325 nm for LDPCL-CHD 1%, 3%, and 5%, respectively. The hydrophilic properties of the nanofibers were significantly improved by incorporating CHD. Also, the contact angle was reduced from 116 ± 5° to 37 ± 2°. The CHD was released from the nanofibers by an initial immediate release and then by extended-release. The released CHD from the nanofibers was found to effectively reduce the formation of biofilms when exposed to microorganisms such as S. aureus and E. coli compared to the control group (LDPCL). The drug-loaded nanofibers were found to be non-cytotoxic when incubated with L929 fibroblast cells for the duration of 24 and 72 h. The study findings suggest that antimicrobial nanofibers have potential applications in preventing implant-related infections caused by (S. aureus and E. coli).

Perinatal health effects of herbicides exposures in the United States: the Heartland Study, a Midwestern birth cohort study

BackgroundThe objective of the Heartland Study is to address major knowledge gaps concerning the health effects of herbicides on maternal and infant health. To achieve this goal, a two-phased, prospective longitudinal cohort study is being conducted. Phase 1 is designed to evaluate associations between biomarkers of herbicide concentration and pregnancy/childbirth outcomes. Phase 2 is designed to evaluate potential associations between herbicide biomarkers and early childhood neurological development.MethodsPeople (target enrollment of 2,000) who are seeking prenatal care, are ages 18 or older, and are ≤ 20 + 6 weeks gestation will be eligible for recruitment. The Heartland Study will utilize a combination of questionnaire data and biospecimen collections to meet the study objectives. One prenatal urine and buccal sample will be collected per trimester to assess the impact of herbicide concentration levels on pregnancy outcomes. Infant buccal specimens will be collected post-delivery. All questionnaires will be collected by trained study staff and clinic staff will remain blinded to all individual level research data. All data will be stored in a secure REDCap database.Hospitals in the agriculturally intensive states in the Midwestern region will be recruited as study sites. Currently participating clinical sites include Indiana University School of Medicine- affiliated Hospitals in Indianapolis, Indiana; Franciscan Health Center in Indianapolis, Indiana; Gundersen Lutheran Medical Center in La Crosse, Wisconsin, and University of Iowa in Iowa City, Iowa. An anticipated 30% of the total enrollment will be recruited from rural areas to evaluate herbicide concentrations among those pregnant people residing in the rural Midwest.Perinatal outcomes (e.g. birth outcomes, preterm birth, preeclampsia, etc.) will be extracted by trained study teams and analyzed for their relationship to herbicide concentration levels using appropriate multivariable models.DiscussionThough decades of study have shown that environmental chemicals may have important impacts on the health of parents and infants, there is a paucity of prospective longitudinal data on reproductive impacts of herbicides. The recent, rapid increases in herbicide use across agricultural regions of the United States necessitate further research into the human health effects of these chemicals, particularly in pregnant people. The Heartland Study provides an invaluable opportunity to evaluate health impacts of herbicides during pregnancy and beyond.Trial registrationThe study is registered at clinicaltrials.gov, NCT05492708 with initial registration and release 05 August, 2022.

Structural Diversity in Antiosteolytic Bisphosphonates: Deciphering Structure-Activity Trends in Ultra Long Controlled Release Phenomena.

Bisphosphonate (BP)-based treatments have been extensively prescribed for bone-related conditions, particularly for osteoporosis. Their low bioavailability creates the need for prescribed dosage increase to reach therapeutic levels but generates a plethora of undesirable side effects. A viable approach to alleviating these issues is to design and exploit controlled release strategies. Herein, the controlled release profiles of 15 structurally characterized BPs (actual drugs and structural analogs) were thoroughly studied from tablets containing three (cellulose, lactose, and silica) or two (cellulose, and silica) excipients in human stomach-simulated pH conditions. The BPs were of two types, alkyl-BPs and amino-BPs. Alkyl-BPs included four derivatives of etidronate (acid, disodium, tetra-sodium, and monopotassium forms), medronic acid, and three analogs of etidronate, in which the -CH3 group was replaced by the moieties -H, -CH2CH2CH3, and -CH2CH2CH2CH2CH3. Amino-BPs included the commercial drugs pamidronate, alendronate, neridronate, and ibandronate, as well as three analog compounds. Release curves were constructed based on data taken from 1H NMR peak integration and were expressed as "% BP release" vs time. The controlled release profiles (initial release rate, plateau value, etc.) were correlated with certain structural features (number of hydrogen and metal-oxygen bonds), showing that the molecular and crystal lattice features of each BP profoundly influence its release characteristics. It was concluded that for all BPs, in general, the initial rate became lower as the total number of lattice interactions increased. For the alkyl-BPs elongation of the alkyl side chain seems to decelerate the release. Amino-BPs, in general, show slower release than the alkyl-BPs. No adverse effects of alkyl- and amino-BP drugs on NIH3T3 cell viability were noted.

Nanocomposite of starch, gelatin and itaconic acid-based biodegradable hydrogel and ZnO/cellulose nanofiber: A pH-sensitive sustained drug delivery vehicle

In recent years, hydrogels as drug carriers have been receiving great interest due to their ability to change their behavior in response to one or more external stimuli. However, their initial burst release profile limits their practical applications. Therefore, we prepared a bio-based hydrogel nanocomposite (HNC) using starch, itaconic acid, acrylic acid and gelatin in the presence of CNF/ZnO-based nanohybrid (ZONH) and used it to evaluate the pH-sensitive drug release properties in different pH solutions. The prepared HNCs were analyzed using various spectroscopic and microscopic techniques. The BET analysis and swelling test of the HNC indicated improved porosity and swelling capacity due to the addition of ZONH. From the drug release study, sustained drug release rate was observed at pH 4 than those at pH 7.4 and 9, indicating controlled release as well as pH responsive behavior of the HNC. Moreover, the drug released HNC was reused as a photocatalyst for dye degradation and achieved good degradation (%). The antibacterial activity of ZONH and HNC was observed against EC and SA bacterial strains from the antibacterial test. In summary, the prepared HNC can be considered as a potential sustainable DDS for biomedical applications as well as a photocatalyst for dye contaminated water treatment.

Zero-order Release of Metformin in Polyacrylamide Hydrogel

Water-soluble polymers with a three-dimensional crosslinked network structure, known as a hydrogel, has widespread applications in clinical and experimental medicine as drug carriers. Upon entering the targeted area for treatment, hydrogel loaded with drugs releases them through both diffusion and self-degradation mechanisms. In this study, the drug release process in polyacrylamide hydrogel uniformly loaded with metformin hydrochloride is investigated. To achieve a uniform drug release rate from the hydrogel, we establish a statistical model of drug release from polyacrylamide hydrogel using response surface methodology based on experimental design technique. The established statistical model can predict drug release behavior at a low cost within the design domain. The relationship between initial drug concentration and drug release rate in metformin-loaded polyacrylamide hydrogel has been investigated. A zero-order drug release effect in hydrogels is achieved through an optimized design of the initial distribution of drug concentration of a five-layered drug-containing hydrogel. This study has significant implications for the realization of drug molecule transport and sustained release in vivo.

Formulation of chitosan based lipid polymer hybrid nanocarrier system for oral delivery of tamoxifen citrate: Toxicological and hemocompatibility evaluation

The objective of the current work was to develop chitosan based lipid nanocarriers for the oral controlled release of tamoxifen citrate (TC). The ionic gelation method was used to develop various formulations with altered ratios of polymer, lipid, and emulsifier. The developed lipid hybrid nanoparticles (LHNPs) were characterized for particle size, polydispersity index, zeta potential, entrapment efficiency, SEM, DSC, FTIR, PXRD, in vitro release studies, stability studies, kinetic modeling, toxicity studies, and hemocompatibility assay. Particle size and entrapment efficiency of the developed formulations varied between 138.2 ± 4.2 nm to 281.8 ± 63.1 nm and 70.94 ± 0.29 % to 82.23 ± 0.02 %, respectively. The polydispersity index and zeta potential ranged from 0.199 ± 0.03 to 0.555 ± 0.01 and 17.9 ± 0.62 to 32.3 ± 0.36, respectively. Nano-sized, spherical in shape surfaces of developed particles were depicted by SEM. While removal of TC melting endotherm revealed by DSC analysis, suggested good thermal stability of LHNPs. FTIR verified that the components of the formulation do not interact chemically. PXRD depicted that the developed formulation has been successfully amorphized. In vitro release study revealed drug release, an initial burst drug release followed by the controlled release up to 72 h. Kinetic modeling exposed that all developed formulations showed a non-Fickian diffusion profile and were best fitted into Higuchi's square root and Korsmeyer-Peppas models. Toxicity investigations on female albino rabbits provided evidence for the compatibility and non-toxicity of developed particles with biological systems. Hemocompatibility assay showed that the developed system is compatible with blood and nontoxic. These investigations demonstrate the potential of developed LHNPs as a vehicle, aimed at controlled oral delivery of TC for better therapeutic applications in breast cancer.

Mechanistic analysis of the photolytic decomposition of solid-state S-nitroso-N-acetylpenicillamine

S-Nitroso-N-acetylpenicillamine (SNAP) is among the most common nitric oxide (NO)-donor molecules and its solid-state photolytic decomposition has potential for inhaled nitric oxide (iNO) therapy. The photochemical NO release kinetics and mechanism were investigated by exposing solid-state SNAP to a narrow-band LED as a function of nominal wavelength and intensity of incident light. The photolytic efficiency, decomposition products, and the photolytic pathways of the SNAP were examined. The maximum light penetration depth through the solid layer of SNAP was determined by an optical microscope and found to be within 100–200 μm, depending on the wavelength of light. The photolysis of solid-state SNAP to generate NO along with the stable thiyl (RS·) radical was confirmed using Electron Spin Resonance (ESR) spectroscopy. The fate of the RS· radical in the solid phase was studied both in the presence and absence of O2 using NMR, IR, ESR, and UPLC-MS. The changes in the morphology of SNAP due to its photolysis were examined using PXRD and SEM. The stable thiyl radical formed from the photolysis of solid SNAP was found to be reactive with another adjacent thiyl radical to form a disulfide (RSSR) or with oxygen to form various sulfonyl and sulfonyl peroxyl radicals {RS(O)xO·, x = 0 to 7}. However, the thiyl radical did not recombine with NO to reform the SNAP. From the PXRD data, it was found that the SNAP loses its crystallinity by generating the NO after photolysis. The initial release of NO during photolysis was increased with increased intensity of light, whereas the maximum light penetration depth was unaffected by light intensity. The knowledge gained about the photochemical reactions of SNAP may provide important insight in designing portable photoinduced NO-releasing devices for iNO therapy.

Experimental investigation of hydrogen jet flame inhibition by nitrogen jet

With the rapid development of hydrogen energy and the increase of the risk of hydrogen jet fires, a reliable fire protection technology is required to extinguish hydrogen jet flame efficiently. An experimental investigation was carried out to study the inhibition of a hydrogen jet flame by a nitrogen jet. The impact angle, nozzle diameter and nozzle position of the nitrogen jet were the variables. The flame characteristic parameters were analyzed, including the flame length, deflection height and deflection angle. The results show that the inhibited flame length was always positively correlated with the vertical distance between the nozzles. The increase of the impact angle had a significant inhibitory effect on the flame length. Besides, the explanatory factor was used to describe the influence of various initial release conditions on flame inhibition and was calculated by the combination of the explanatory variables, such as impact angle, impact stroke and impact height. The flame length reduction ratio, deflection height reduction ratio and dimensionless deflection angle are positively correlated with the explanatory factor. It means that the jet flame is more easily extinguished under a larger explanatory factor. The critical parameter of the explanatory factor for extinguishing the hydrogen jet flame was verified experimentally.

Relational Reprojection Platform: Non-linear distance transformations of spatial data in R

When mapping relationships across multiple spatial scales, prevailing visualization techniques treat every mile of distance equally, which may not be appropriate for studying phenomena with long-tail distributions of distances from a common point of reference (e.g., retail customer locations, remittance flows, and migration data). While quantitative geography has long acknowledged that non-Cartesian spaces and distances are often more appropriate for analyzing and visualizing real-world data and complex spatial phenomena, commonly available GIS software solutions make working with non-linear distances extremely difficult. Our Relational Reprojection Platform (RRP) fills this gap with a simple stereographic projection engine centering any given data point to the rest of the set, and transforming great circle distances from this point to the other locations using a set of broadly applicable non-linear functions as options. This method of reprojecting data allows users to quickly and easily explore how non-linear distance transformations (including square root and logarithmic reprojections) reveal more complex spatial patterns within datasets than standard projections allow. Our initial release allows users to upload comma separated value (CSV) files with geographic coordinates and data columns and minimal cleaning and explore a variety of spatial transformations of their data. We hope this heuristic tool will enhance the exploratory stages of social research using spatial data.

Aqueous remote loading of setmelanotide in poly(lactic-co-glycolic acid) microspheres for long-term obesity treatment

Setmelanotide (Imcivree™) was developed as a daily injectable therapeutic peptide for the treatment of rare forms of syndromic obesity, such as POMC deficiency and leptin receptor deficiency. The important option of poly(lactic-co-glycolic acid) (PLGA) controlled release microspheres has become more attractive for this class of drugs upon the discovery that net positively charged peptides can be remote-loaded rapidly from aqueous peptide solution into blank microspheres at high loading and encapsulation efficiency. Here we sought to remote-load setmelanotide in PLGA microspheres and examine its potential for long-term controlled release and body weight control. The influence of PLGA microsphere porosity was investigated with respect to morphology, drug loading, and in vitro release profiles. Increased density of the microspheres inhibited the progress of encapsulation of the dicationic peptide. A diet-induced obese murine model was then used to determine the pharmacokinetic profile and to evaluate long-term efficacy of an optimal formulation. Remote loaded PLGA formulations encapsulated setmelanotide as high as ∼63% (∼6.3% w/w loading) and exhibited slow and continuous peptide release over ∼6 weeks in vitro largely independent of microsphere porosity. The obtained in vivo release pattern from deconvolution of the pharmacokinetics after subcutaneous microsphere injection was consistent with the in vitro release profile but with a lower initial burst release and overall slightly faster release rate. After a single injection of remote-loaded setmelanotide, continuous long-term inhibition of food intake and body weight control was observed over 17 and 30 days, respectively. The improvement in body weight control over drug-free microsphere vehicle-treated control groups matched the observed PK profile. This study provides the first report of long-acting release formulation for 1-month controlled release of setmelanotide and body weight control in a diet induced obese murine model, and supports the further development of long-acting treatment options for obese patients.

Docetaxel-Loaded Methoxy poly(ethylene glycol)-poly (L-lactic Acid) Nanoparticles for Breast Cancer: Synthesis, Characterization, Method Validation, and Cytotoxicity.

This study aimed to synthesize and characterize DTX-mPEG-PLA-NPs along with the development and validation of a simple, accurate, and reproducible method for the determination and quantification of DTX in mPEG-PLA-NPs. The prepared NPs were characterized using AFM, DLS, zetasizer, and drug release kinetic profiling. The RP-HPLC assay was developed for DTX detection. The cytotoxicity and anti-clonogenic effects were estimated using MTT and clonogenic assays, respectively, using both MCF-7 and MDA-MB-231 cell lines in a 2D and 3D culture system. The developed method showed a linear response, high precision, accuracy, RSD values of ≤2%, and a tailing factor ≤2, per ICH guidelines. The DTX-mPEG-PLA-NPs exhibited an average particle size of 264.3 nm with an encapsulation efficiency of 62.22%. The in vitro drug kinetic profile, as per the Krosmeyers-Peppas model, demonstrated Fickian diffusion, with initial biphasic release and a multistep sustained release over 190 h. The MTT assay revealed improved in vitro cytotoxicity against MCF-7 and MDA-MB-231 in the 2D cultures and MCF-7 3D mammosphere cultures. Significant inhibitions of the clonogenic potential of MDA-MB-231 were observed for all concentrations of DTX-mPEG-PLA-NPs. Our results highlight the feasibility of detecting DTX via the robust RP-HPLC method and using DTX-mPEG-PLA-NPs as a perceptible and biocompatible delivery vehicle with greater cytotoxic and anti-clonogenic potential, supporting improved outcomes in BC.

Nanohybrid Based on (Mn, Zn) Ferrite Nanoparticles Functionalized With Chitosan and Sodium Alginate for Loading of Curcumin Against Human Breast Cancer Cells

This study reports on the synthesis of Mn1 − xZnxFe2O4 (Mn, Zn ferrite) magnetic nanoparticles (MNPs) as drug delivery carriers for effective therapeutic outcomes. The MNPs were prepared using the coprecipitation method, and their magnetic properties were investigated based on their composition. Among the compositions tested, Mn0.8Zn0.2Fe2O4 MNPs exhibited superparamagnetic properties with a saturation magnetization moment of 34.6 emu/g at room temperature (25°C). To enhance the water solubility of curcumin (Cur), known for its hydrophobic nature, it was successfully loaded onto alginate (Alg)/chitosan (Chit)@Mn0.8Zn0.2Fe2O4 nanoparticles (NPs). The nanocomposite was characterized by field emission scanning electron microscopy (FE-SEM) which revealed a particle size of approximately 20 nm. The crystalline structure of the NPs was analyzed using X-ray diffraction, while Fourier-transform infrared (FTIR), energy-dispersive X-ray, and map analysis techniques were employed for further characterization. In terms of drug release, there was an initial burst release of Cur (around 18%) within the first hour, followed by a slower release (approximately 61%) over the next 36 h. The anti-tumor properties of the Cur-loaded NPs were evaluated using the Methyl Thiazol Tetrazolium (MTT) assay and quantitative real-time polymerase chain reaction. The MTT assay confirmed a higher cytotoxic effect of Cur-loaded Alg/Chit@Mn0.8Zn0.2Fe2O4 NPs on the MCF-7 breast cancer cell line compared to free Cur, highlighting the significance of incorporating Cur into nano-sized carrier systems.Graphical

Abstract 11891: Pulmonary Delivery of Novel BEPI/NO-Microparticle for Treatment of Pulmonary Arterial Hypertension

Introduction: Pulmonary arterial hypertension (PAH) is an incurable disease characterized by elevated blood pressure in the pulmonary artery causing right heart failure and even sudden cardiac death. Nitric oxide (NO) is a strong potent regulator of PAH. However, the therapeutic effects of exogenous NO depend on its concentration and its half-life is very short. Hypothesis: we prepared NO-releasing inhalers with controlled porous structures to modulate the deposition in lung. Methods and Results: NO-releasing inhalers with micro-sized pores showed improved nebulization efficiency. Furthermore, the cGMP expression and macrophage polarization were evaluated. The fluorescence intensity of Nile red was analyzed using photoluminescence spectroscopy to confirm the amounts of NO-MPs on the slide glass. As with confocal images, 0.8k BPEI/NO-MPs could arrive the lower region than 25k BPEI/NO-MPs. NO produced by endothelial nitric oxide synthase (eNOS) induce vasodilation by increasing cGMP in vascular smooth muscle cells. Hence, the delivery of NO mimicking the physical production of the endothelium in range of 0.05-0.4 nmol cm -2 min -1 is required to restore vascular homeostasis. Therefore, BPEI0.8k/NO-MPs release 0.277 nmol cm -2 min -1 of NO that mimics the environment in healthy endothelium, while BPEI25k/NO-MPs showed a higher concentration as 0.548 nmol cm -2 min -1 . BPEI0.8k/NO-MPs increased the synthesis of cGMP in cells, whereas BPEI25k/NO-MPs presented adverse effects due to an initial burst release. When cell populations were quantified, BPEI0.8k/NO-MPs increased the number of M2-like cells while decreasing the number of M1-like cells. Furthermore, using fluorescence activated cell sorting analysis, proteins and gene levels of cells, anti-inflammatory factors increased in BPEI0.8k/NO-MPs treatment groups Conclusions: novel BPEI/NO-releasing inhalers could deliver NO to deep lungs and has the potential for treatment of PAH

Use of biosilica to improve loading and delivery of bone morphogenic protein 2

The clinical utility of bone morphogenetic protein 2 (BMP2) is limited because of the poor attraction between BMP2 and carriers, resulting in low loading efficiency and initial burst release. Here, the high binding affinity of BMP2 to the biosilica surface was utilized to overcome this limitation. Atomic force microscopy revealed that BMP2 bound nearly 8- and 2-fold more strongly to biosilica-coated hydroxyapatite than to uncoated and plain silica-coated hydroxyapatite, respectively. To achieve controlled release, collagen was introduced between the silica layers on hydroxyapatite, which was optimized by adjusting the collagen concentration and number of layers. The optimal biosilica/collagen formulation induced sustained BMP2 release without compromising loading efficiency. BMP2 combined with the mentioned formulation led to an increase in osteogenesis, as compared to the combination of BMP2 with either biosilica-coated or non-coated hydroxyapatite in vitro. In rat calvarial defect models, the biosilica/collagen-coated hydroxyapatite with 1 μg BMP2 showed 26 % more bone regeneration than the same dose of BMP2-loaded hydroxyapatite and 10.6 % more than hydroxyapatite with 2.5-fold dose of BMP2. Using BMP2 affinity carriers coated with biosilica/collagen allows for more efficacious in situ loading and delivery of BMP2, making them suitable for the clinical application of growth factors through a soaking method.

In Vivo Evaluation of a Gastro-Resistant Enprotect® Capsule under Postprandial Conditions.

Ready-to-fill enteric hard capsule shells are an evolving field of oral drug and nutraceutical products. Lonza Capsugel® Enprotect® capsules were recently proven to provide reliable release in the small intestine after fasted intake, but robustness against postprandial intake needed to be proven. In this study, the capsules were administered to 16 healthy young subjects after intake of a light meal. The Enprotect® capsules were labelled with 5 mg black iron oxide and 25 mg 13C3-caffeine. Magnetic Resonance Imaging was used to identify the localization and visual dispersion of the capsule filling. The salivary appearance of caffeine was considered a second independent and sensitive marker for the initial release. Whereas the fasted gastric residence time of the capsules amounted to 43 ± 32 min, it was increased to 158 ± 36 min after postprandial intake. Therefore, the mean dispersion time according to MRI and the mean caffeine appearance time were increased to 196 ± 37 min and 189 ± 37 min, respectively. But, similar to fasted administration, no capsule disintegration or leakage was observed in the stomach and 38% of the capsules disintegrated in the jejunum and 62% in the ileum. The mean dispersion time after gastric emptying and the mean caffeine appearance time after gastric emptying amounted to 38 ± 21 min and 31 ± 17 min, respectively. Both did not relevantly change compared to the fasted intake. Only the absolute dispersion time and caffeine appearance were prolonged due to the increased gastric residence and no relevant influence of the light meal was observed on the disintegration or release behavior of Enprotect® capsules after gastric emptying. The capsules also showed robust enteric properties after postprandial administration.

AlphaFold Protein Structure Database in 2024: providing structure coverage for over 214 million protein sequences.

The AlphaFold Database Protein Structure Database (AlphaFold DB, https://alphafold.ebi.ac.uk) has significantly impacted structural biology by amassing over 214 million predicted protein structures, expanding from the initial 300k structures released in 2021. Enabled by the groundbreaking AlphaFold2 artificial intelligence (AI) system, the predictions archived in AlphaFold DB have been integrated into primary data resources such as PDB, UniProt, Ensembl, InterProand MobiDB. Our manuscript details subsequent enhancements in data archiving, covering successive releases encompassing model organisms, global health proteomes, Swiss-Prot integration, and a host of curated protein datasets. We detail the data access mechanisms of AlphaFold DB, from direct file access via FTP to advanced queries using Google Cloud Public Datasets and the programmatic access endpoints of the database. We also discuss the improvements and services added since its initial release, including enhancements to the Predicted Aligned Error viewer, customisation options for the 3D viewer, and improvements in the search engine of AlphaFold DB.