Morphological Evaluation Research Articles

Novel application of metabolic imaging of early embryos using a light-sheet on-a-chip device: a proof-of-concept study

Abstract STUDY QUESTION Is it feasible to safely determine metabolic imaging signatures of nicotinamide adenine dinucleotide [NAD(P)H] associated auto-fluorescence in early embryos using a light-sheet on-a-chip approach? SUMMARY ANSWER We developed an optofluidic device capable of obtaining high-resolution 3D images of the NAD(P)H autofluorescence of live mouse embryos using a light-sheet on-a-chip device as a proof-of-concept. WHAT IS KNOWN ALREADY Selecting the most suitable embryos for implantation and subsequent healthy live birth is crucial to the success rate of assisted reproduction and offspring health. Besides morphological evaluation using optical microscopy, a promising alternative is the non-invasive imaging of live embryos to establish metabolic activity performance. Indeed, in recent years, metabolic imaging has been investigated using highly advanced microscopy technologies such as fluorescence-lifetime imaging and hyperspectral microscopy. STUDY DESIGN, SIZE, DURATION The potential safety of the system was investigated by assessing the development and viability of live embryos after embryo culture for 67 h post metabolic imaging at the two-cell embryo stage (n = 115), including a control for culture conditions and sham controls (system non-illuminated). Embryo quality of developed blastocysts was assessed by immunocytochemistry to quantify trophectoderm and inner mass cells (n = 75). Furthermore, inhibition of metabolic activity (FK866 inhibitor) during embryo culture was also assessed (n = 18). PARTICIPANTS/MATERIALS, SETTING, METHODS The microstructures were fabricated following a standard UV-photolithography process integrating light-sheet fluorescence microscopy into a microfluidic system, including on-chip micro-lenses to generate a light-sheet at the centre of a microchannel. Super-ovulated F1 (CBA/C57Bl6) mice were used to produce two-cell embryos and embryo culture experiments. Blastocyst formation rates and embryo quality (immunocytochemistry) were compared between the study groups. A convolutional neural network (ResNet 34) model using metabolic images was also trained. MAIN RESULTS AND THE ROLE OF CHANCE The optofluidic device was capable of obtaining high-resolution 3D images of live mouse embryos that can be linked to their metabolic activity. The system’s design allowed continuous tracking of the embryo location, including high control displacement through the light-sheet and fast imaging of the embryos (<2 s), while keeping a low dose of light exposure (16 J · cm−2 and 8 J · cm−2). Optimum settings for keeping sample viability showed that a modest light dosage was capable of obtaining 30 times higher signal-noise-ratio images than images obtained with a confocal system (P < 0.00001; t-test). The results showed no significant differences between the control, illuminated and non-illuminated embryos (sham control) for embryo development as well as embryo quality at the blastocyst stage (P > 0.05; Yate’s chi-squared test). Additionally, embryos with inhibited metabolic activity showed a decreased blastocyst formation rate of 22.2% compared to controls, as well as a 47% reduction in metabolic activity measured by metabolic imaging (P < 0.0001; t-test). This indicates that the optofluidic device was capable of producing metabolic images of live embryos by measuring NAD(P)H autofluorescence, allowing a novel and affordable approach. The obtained metabolic images of two-cell embryos predicted blastocyst formation with an AUC of 0.974. LARGE SCALE DATA N/A. LIMITATIONS, REASONS FOR CAUTION The study was conducted using a mouse model focused on early embryo development assessing illumination at the two-cell stage. Further safety studies are required to assess the safety and use of 405 nm light at the blastocyst stage by investigating any potential negative impact on live birth rates, offspring health, aneuploidy rates, mutational load, changes in gene expression, and/or effects on epigenome stability in newborns. WIDER IMPLICATIONS OF THE FINDINGS This light-sheet on-a-chip approach is novel and after rigorous safety studies and a roadmap for technology development, potential future applications could be developed for ART. The overall cost-efficient fabrication of the device will facilitate scalability and integration into future devices if full-safety application is demonstrated. STUDY FUNDING/COMPETING INTEREST(S) This work was partially supported by an Ideas Grant (no 2004126) from the National Health and Medical Research Council (NHMRC), by the Education Program in Reproduction and Development (EPRD), Department Obstetrics and Gynaecology, Monash University, and by the Department of Mechanical and Aerospace Engineering, Faculty of Engineering, Monash University. The authors E.V-O, R.N., V.J.C., A.N., and F.H. have applied for a patent on the topic of this technology (PCT/AU2023/051132). The remaining authors have nothing to disclose.

Understanding Hemodynamic Incoherence: Mechanisms, Phenotypes, and Implications for Treatment

Abstract The reversal of microcirculation dysfunction is crucial for assessing the success of shock resuscitation and significantly influences patient prognosis. However hemodynamic incoherence is observed when microcirculatory dysfunction persists despite the restoration of macrocirculatory function post-resuscitation. Recent advancements in technology have enabled bedside assessment of microcirculation in shock patients, allowing for direct visualization of microcirculatory morphology and quantitative evaluation of its functional status. This article reviews the pathophysiological mechanisms that lead to hemodynamic incoherence. It also introduces the current understanding and classification framework for the different phenotypes of hemodynamic incoherence. Existing evidence indicates that the diverse mechanisms leading to microcirculatory disorders result in varied manifestations among patients experiencing hemodynamic incoherence, highlighting the heterogeneity of this population. Some classification frameworks have been proposed to enhance our understanding of these phenotypes. By integrating pathophysiological mechanisms, clinical symptoms, indicators of macrocirculation, microcirculation, tissue metabolism, and biomarkers, we can summarize certain clinical features of phenotypes in hemodynamic incoherence to form a conceptual framework. Additionally, strategies for creating targeted treatments based on different phenotypes require further validation.

PVDF‐based nanocomposite coatings with superhydrophobic and antibacterial properties on stainless steel surfaces

AbstractIn this study, using a spray‐coating technique, stainless steel surfaces were coated with polyvinylidene fluoride (PVDF) and carbon soot nanoparticles. We imparted various surface properties onto the coatings' surfaces by varying the nanoparticle content and the preparation method. The main aim was to develop antibacterial surfaces by inducing superhydrophobic properties in stainless steel surfaces. In a one‐step preparation method, 5% and 10% of nanoparticles were insufficient to achieve superhydrophobicity, whereas 20% significantly increased the water contact angle to 153°. Morphological evaluations revealed the formation of a nano‐scale structure, which also led to a staggering antibacterial rate of 99.5% against Listeria monocytogenes. Using a two‐step preparation, soot nanoparticles were localized at the coating's top layer, which led to superhydrophobicity at a lower nanoparticle content (10%). However, the bacterial adhesion study showed a lower antibacterial rate (~93%). The difference in bacterial adhesion results was explained by various roughness structures according to the atomic force microscopy results. The developed system has a promising potential to be used as self‐cleaning and antibacterial coatings in the medical and food industries.Highlights Stainless steel surfaces were coated with PVDF and carbon soot nanoparticles Imparted various surface properties onto the coatings' surfaces Developed antibacterial surfaces by inducing superhydrophobic properties Using a two‐step, soot nanoparticles were localized at the coating's top layer Bacterial adhesion study showed a lower antibacterial rate (~93%).

MORPHOLOGICAL CHARACTERIZATION OF PIGEON PEA [Cajanus cajan (L.) Millspaugh] ACCESSIONS FOR AGRONOMIC TRAITS IN ASSOSA DISTRICT, NORTHWESTERN ETHIOPIA

<p><strong>Background:</strong> Pigeon pea is a tropical grain legume being cultivated as important pulses for human food, feed and soil fertility improvement. Pigeon pea research in terms of crop improvement is still at a low level in Ethiopia. <strong>Objective: </strong>To estimate the Genetic variability of pigeon pea accessions with respect to important qualitative and quantitative traits and select high yielding grain and promising traits for variety development. <strong>Methodology</strong>: Treatments consisted in twenty two pigeon pea accessions (MPG1, MPG2, MPG3, MPG4, MPG5, MPG6, MPG7, MPG8, MPG9, MPG10, MPG11, MPG12, MPG13, MPG14, MPG15, MPG16, MPG17, MPG18, MPG19, MPG20, MPG21 and MPG22) collected from Assosa area. Treatments were laid out in a Randomized Complete Block Design with three replications. <strong>Results:</strong> Qualitative characteristics recorded for accessions showed different modalities and frequencies at vegetative growth stage with the qualitative traits dominated were semi-spreading growth habit 65 %, leaflet shape with lanceolate 70 %, color about 44 % of the accessions exhibited yellow, seed size where 55 % accounted large sized, seed color pattern: 52% of accessions showed plain, pod color where 56 % genotypes had purple, seed with red dominated was 50 % and 71 % of accessions had totally pigmented pod. With respect to quantitative traits, genotypes accessions MPG11 and MPG15 had relatively higher grain yield (> 4000 kg/ ha) with the highest grain yield recorded for genotype MPG18. <strong>Implications:</strong> The investigation of the present study for morphological evaluation of genotype Pigeon pea exhibited genetic variability among accession. <strong>Conclusion</strong>: Agronomic traits such as plant height, branches per plant, pods per plant, seeds per pod, pod length, pod width, hundred seed weight and grain yield for accessions were exhibited significant differences which suggest high yielding accessions could be directly recommended for production while others could be utilized effectively to develop high yielding varieties through hybridization followed by selection. </p>

Navigating the Diagnostic Challenges in Lymph Node Cytology: The Case of Reactive Hyperplasia.

Fine needle cytology (FNC) is a pivotal diagnostic tool for distinguishing between benign and malignant lymphadenopathies mainly because of its minimal invasiveness, cost-effectiveness and accuracy. A major requirement for maximising diagnostic accuracy is proper sample management of aspirated cellular material. In this diagnostic process, the morphological evaluation of adequate smears is paramount, guiding cytopathologists in the selection of appropriate ancillary tests through the recognition of cell size and patterns of distribution. Here, we describe a peculiar 'concentric ovals distribution pattern', frequently observed in the FNC of benign reactive lymph nodes, which may represent an aid in the cytological diagnosis of reactive hyperplasia.

Ferric ions crosslinked hyaluronic acid beads: potentials for drug delivery use

Introduction and purpose Despite the attractive properties of hyaluronic acid (HA), The preparation of HA beads is still challenging. This article reports the preparation of pH-sensitive gel HA beads. The ionic gelation method was used to prepare the HA gel beads using ferric ions. This cross-linking type is based on forming coordination bonds, which enhance the mechanical properties of the prepared beads. Methods The developed beads were characterized using Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). Scanning electron microscopy (SEM) examined the bead’s morphology. Furthermore, the potential of HA gel beads as an oral drug delivery system was investigated using metformin as a hydrophilic model drug. The entrapment efficiency and in vitro, release, and release kinetics were evaluated. The crosslinking density and HA concentration effect on drug release and bead swelling capacity under pH 1.2 and 7.4 were also investigated. Results The entrapment efficiency of metformin in HA beads was found to be 79.56 ± 3.89%. FTIR analysis indicated the ionic interaction between ferric ions and the carboxylic groups on the HA molecule. At the same time, there was no substantial interaction between metformin and the polymeric bead. Morphological evaluation and DSC analysis suggested the successful incorporation of metformin within the beads. The in vitro drug release evaluation showed pH-dependent extended release where the release kinetics followed the first-order mathematical model. Conclusions This study provides a value-added formulation with the potential for drug delivery use, which can be further investigated for biomedical applications.

Disintegration of commercial single-use plastics from synthetic and biobased origins and effects on plant growth

Many single–use plastic (SUP) options made of synthetic polymers, bio-based materials, and blends of both are available in the market and used in large quantities. The disintegration of eleven commercial SUP, marketed in Mexico as cups and plates, was investigated in an aerobic home compost environment at a laboratory scale over 180 days. An evaluation of chemical changes, surface morphology, and thermal and mechanical properties was conducted to ascertain the original composition of SUP and the progression of disintegration in samples that are challenging to clean from soil contamination. Furthermore, the impact of residual compost on barley (Hordeum vulgare) plant growth and its correlation with the leaching of heavy metals were explored. The bio-based SUP, but not those made of expanded polystyrene foam, showed a correlation between the disintegration degree (measured by weight loss into particles <2 mm) and a decrease in functional groups (observed by FT-IR), mechanical-thermal stability loss, and surface wear over disintegration time. For instance, the highest disintegration at 180 days was approximately 70% for wheat bran and palm leaf plates, followed by wheat plates and cellulose-PLA cups (60%). In addition to the components listed by the manufacturers, the FT-IR and DSC analysis revealed the presence of polyethylene and polypropylene in cellulose cups and sugarcane plates. These components, impede disintegration but contribute to preserving thermal resistance and hydrophobicity during utilization. Compost derived from expanded polystyrene foam SUP, with 90 days of disintegration, was rich in zinc and chromium and significantly decrease in the root length of the barley plant compared to the control. This demonstrates the necessity of considering the impact of the leaching of additives and secondary microplastics into the environment.

IMPACT OF BLASTOCYST GRADES TO IMPLANTATION AND LIVE-BIRTH RATES IN WOMEN LESS THAN 30 YEARS WITH SINGLE EMBRYO TRANSFERS

Background:Morphological evaluation of blastocysts focusing on key factors such as expansion, inner cell mass (ICM) quality, and trophectoderm (TE) quality have been most widely used for embryo selection. The study emphasizes the importance of detailed blastocyst evaluation, highlighting that morphological variations can have substantial implications for ART outcomes. Methods:A total of 115women aged less than 30 years undergoing SET from from January 2023 to June 2023 were included in this study.The patients were divided into three groups based on their morphologic grading before embryo transfer: good-quality (n = 53), average-quality (n = 52) and poor-quality blastocysts (n = 10). The three morphologic groups pregnancy outcomes were examined, and the implantation rate was logistically regressed. Results:Good-quality blastocysts yielded a statistically significantly higher implantation rate (77%) than average / poor-quality blastocysts (44% and 20% respectively). Average-quality blastocysts still yielded in higher implantation rates (44%) compared to poor-quality blastocysts (20%). The baseline characteristics were analysed for significance using the Pearsons Chi-squared test. A P-value &lt; 0.05 was considered statistically significant. All the statistical analyses were performed using the social science statistics website with SPSS software. Conclusions:Higher blastocyst morphologic grading was associated with increased implantation rate for single embryo transfers. These findings suggest that evaluating blastocyst morphology is critical when selecting the best blastocyst.

Quantitative characterisation of debris cloud-induced pitting damage in AL-Whipple shields of spacecraft using infrared thermography

ABSTRACT Quantitatively evaluating the debris cloud-induced pitting damage in the AL-Whipple structure of spacecraft, is still a challenge, let alone microstructural plastic deformation usually accompanied by geometric cratering defects, which may decrease material stress-carrying capacity and structural life. With this motivation, a dedicated modelling technique is proposed to gain an insight into various modalities (i.e., geometric and plastic defects) of pitting damage-induced thermal modulation for infrared inspection. Then, a sequence of infrared images is obtained and analysed using the gap smoothing algorithm and naive Bayes classification methods. On this basis, a quantitative characterization framework is proposed to characterize the damage modalities and their morphologies. This method is validated using a 3D numerical model which contains pitting damage acquired from the real-world structure. Modality classification and morphology evaluation results are well consistent with the artificial defects, in terms of the modality, diameter, and depth. Then they‘re experimentally corroborated, in which the severity and distribution of pitting damage are precisely characterized, in terms of the crater number and morphologies, as well as the size of plastic regions around these craters, which are consistent with the actual observations. This work proposes a non-destructive strategy for pitting damage evaluation, which is engineering-friendly.

Predictor of A4-amplitude using preprocedural electrocardiography in patients with leadless pacemakers

Abstract Background Based on the historical studies of leadless pacemakers (LP), high atrio-ventricular synchrony (AVS) with mechanical sensing-based VDD pacing is largely influenced by A4-amplitude. A limited study investigated the predictors of A4-amplitude using clinical and echocardiographic parameters. Objective This study aimed to investigate the predictors of A4-amplitude preoperatively to select patients who could benefit the most from AVS among patients with VDD LPs. Methods Data from patients who received VDD LP implantations from November 2021 to August 2023 at our institution were analyzed. Twelve-lead electrocardiography and transthoracic echocardiography were performed before VDD LP implantations. To assess the electrical indices associated with the A4-signal, the electrocardiographic morphological P-wave parameters were analyzed including the P-wave duration, P-wave amplitude, maximum deflection index (MDI), and P-wave dispersion. Results A total of 50 patients who underwent VDD LP implantations (median age, 84 years; 64% male) were included and divided into two groups based on the median value of A4-amplitude, the high-A4 group (A4-amplitude &amp;gt;2.5m/s2, n=26) and low-A4 group (A4-amplitude ≤2.5m/s2, n=24). There was a significant difference between the high-A4 and low-A4 groups in the LVEF (P=0.01), P-wave dispersion (P=0.01), and MDI (P&amp;lt;0.001). A multivariate logistic analysis revealed that a lower MDI was an independent predictor of a high A4-amplitude (odds ratio: 0.78, 95% confidence interval: 0.67 to 0.92; P=0.003). Conclusion Preoperative electrocardiographic evaluations of the P-wave morphology may be useful for predicting A4-amplitude. The MDI was the only independent A4-amplitude predictor that seemed promising for selecting VDD LP patients.abstract figure

Myocardial determinants of Functional limitation in advanced Fabry cardiomyopathy

Abstract Background Cardiac involvement in Fabry Disease (FD) manifests as left ventricular hypertrophy (LVH) often complicated by myocardial fibrosis and/or inflammation. Both LVH and advanced tissue alterations can be non-invasively detected and quantified by cardiac magnetic resonance (CMR) with Late Gadolinium Enhancement (LGE). Impairment in functional capacity has been previously reported in FD patients. However, there is limited data regarding the determinants of myocardial alterations in exercise intolerance in FD. Methods Among 190 consecutive FD patients referred for CMR, 41 simultaneously performed a cardiopulmonary exercise test (CPET) to evaluate functional capacity. All patients underwent an extensive CMR protocol to provide a detailed evaluation of cardiac morphology and function, including multiparametric tissue characterization. CPET parameters were compared between patients with advanced Fabry cardiomyopathy (Group B), characterized by hypertrophy, inflammation, and fibrosis, and FD patients without irreversible cardiac damage (Group A). Results Group B patients were older compared to Group A (57.0 years vs 33.5 years, p&amp;lt;0.001), had a greater LVH (LVMI: 117 g/sqm vs 76.5 g/sqm, p&amp;lt;0.001; LVMWT 17 vs 10 mm, p&amp;lt;0.001). LV ejection fraction was preserved in both groups but Group B patients had a lower peak VO2 value compared to group A (17.8 ml/kg/min vs 21.9 ml/kg/min, p=0.025), a lower (but not significant) percent-predicted peak VO2. A negative linear correlation was identified between the LGE extension (expressed as % of LV mass) and peak VO2 (r=-0.553, p=0.014). This correlation remained consistent even after excluding patients under chronic beta-blocker therapy (r=-0.678, p=0.045). No significant correlation between LV mass and peak VO2 was found. Conclusions in patients with FD, the presence of advanced tissue alterations detected by LGE is associated with reduced functional capacity (low peak VO2) despite preserved LV ejection fraction, with a significant negative correlation.

Segmental post-PCI physiological gradients using ultrasonic or optical flow ratio

Abstract Background Segmental pressure gradients post percutaneous coronary intervention (PCI) may detect residual disease and potential targets for optimization. Ultrasonic flow ratio (UFR) or optical flow ratio (OFR) provide simultaneous physiological and morphological data using a single imaging catheter. Notably, both have been validated against wire based FFR Objectives To evaluate the utility of UFR and OFR to identify residual disease post-PCI. Methods The present study is a sub analysis of the ASET (Acetyl Salicylic Elimination Trial) JAPAN pilot study Phase I which showed the feasibility and safety of low dose prasugrel monotherapy following PCI in Japanese patients presenting with chronic coronary syndrome and an anatomical SYNTAX score &amp;lt;23. Patients with complete intra-vascular imaging pullback data were included with UFR/OFR acquired in all patients post-PCI. Quantitative and qualitative assessment of IVUS or OCT pullbacks was performed by automatically delineating vessel, lumen, and stent contours every 0.5 mm using QCU-CMS (version 4.69, Division of Image Processing, Leiden University Medical Centre). UFR or OFR was calculated using the respective IvusPlus or OctPlus software, Pulse Medical, Shanghai, China.(13)(15) All UFR and OFR tracings, and IVUS and OCT pullbacks were analyzed at the CORRIB Core lab, Galway, Ireland by two analysts blinded to clinical and procedural data. Anatomically focal lesions distal and proximal to the stent were evaluated in segments ≥5mm long. UFR/OFR virtual pullback curves were used to calculate intra-stent pressure gradients and physiologically define lesions as focal or diffuse by segmental pressure drops ≥0.05 over lengths &amp;lt;10mm or ≥10mm, respectively. Results The median post-PCI UFR/OFR was 0.93 (0.88-0.96) with 69/195 vessels (35.4%) having a UFR/OFR≤0.90. There were significantly more focal anatomical and physiological lesions proximal and distal to the stent in vessels with a UFR/OFR≤0.90 versus &amp;gt;0.90. In vessels with a UFR/OFR≤0.90 there was moderate agreement (weighted kappa=0.553, p&amp;lt;0.001) between anatomical and physiological focal lesions proximal to the stent, whilst it was fair (weighted kappa=0.219, p=0.002) distally. In-stent gradient had poor discriminative ability to detect significant stent under-expansion. The performance of a proximal or distal virtual FFR gradient to detect proximal or distal anatomically focal disease was good (AUC–0.835 and 0.877, respectively) Conclusions UFR/OFR enables suboptimal vessel physiology post-PCI to be detected and helps identify the precise anatomical location with corroboration from intra-vascular imaging. Adequately powered physiology optimization trials using UFR/OFR will determine the role of combined morphology and physiological evaluation post PCI in improving clinical outcomes.

Acid-etching protocol for bioceramic cements: Evaluation of bond strength, compression and morphology.

To establish an acid-etching protocol for Biodentine and Cimmo DTA, evaluating compressive strength, bond strength, surface morphology in scanning electron microscope and failure modes after different etching times. Two test specimens were prepared for each cement and divided into four groups (n = 12) according to the acid-etching time (0, 5, 10 and 15 s). Compressive strength was tested using a universal testing machine, while bond strength was evaluated after bonding with Filtek Bulk Flow resin using Universal ESPE Single Bond adhesive. Failures were classified as surface-adhesive, cement-cohesive, resin-cohesive and mixed. Biodentine showed significantly higher compressive strength than Cimmo DTA (p < 0.001), regardless of acid etch time (p < 0.001). Different acid-etching strategies are required for Biodentine and Cimmo DTA, with Biodentine requiring selective etching and Cimmo DTA requiring a full 15-s etch to optimise bond strength properties.

Diagnostic accuracy of bone marrow blood evaluation in haemophagocytic lymphohistiocytosis paediatric patients.

Haemophagocytic lymphohistiocytosis (HLH) is a rare and serious immunological syndrome that involves a strong activation of cytotoxic T lymphocytes and macrophages. HLH determines a cytokine-mediated tissue injury with a contemporary multi-organ failure and a high fatality rate. A retrospective study was performed considering the medical records of paediatric patients who underwent a bone marrow aspirate for suspect HLH. The biomarkers evaluated were among those included in the HLH-2004. Lactate dehydrogenase (LD) was also evaluated. Haemophagocytosis was evaluated in bone marrow blood smear slides. Enrolled were 11 patients included in the HLH group and 8 patients as controls. Haemoglobin and fibrinogen resulted lower in HLH patients than in controls, while blood triglycerides, serum ferritin and LD resulted increased. Blood triglycerides and fibrinogen discriminated HLH cases perfectly, with a sensitivity and specificity of 100%. Ferritin had a sensitivity of 100% and a specificity of 83% (cut off ≥3,721µg/L) and LD of 73% and of 100% (the cut off ≥1,903U/L). Haemoglobin was found to have a sensitivity of 75% and a specificity of 100% (cut off ≤ 96g/L). Total haemophagocytes cell counts were not different between patients and controls. Only the increased number of phagocytized nucleated red blood cells (NRBC) was found to be significantly increased in the patients. Erythrocytes phagocytosis (≥4/1,000 cells) only tended towards significance. The blood biomarkers showed better diagnostic performance than the morphological evaluation. Among the different cell lineages engulfed by haemophagocytes, the best diagnostic performance was obtained by phagocytosed mature erythrocytes and immature nucleated erythrocytes.

Double-Network Hydrogel 3D BioPrinting Biocompatible with Fibroblast Cells for Tissue Engineering Applications

The present study examines the formulation of a biocompatible hydrogel bioink for 3D bioprinting, integrating poly(ethylene glycol) diacrylate (PEGDA) and sodium alginate (SA) using a double-network approach. These materials were chosen for their synergistic qualities, with PEGDA contributing to mechanical integrity and SA ensuring biocompatibility. Fibroblast cells were included in the bioink and printed with a Reg4Life bioprinter employing micro-extrusion technology. The optimisation of printing parameters included needle size and flow velocities. This led to precise structure development and yielded results with a negligible deviation in printed angles and better control of line widths. The rheological characteristics of the bioink were evaluated, demonstrating appropriate viscosity and shear-thinning behaviour for efficient extrusion. The mechanical characterisation revealed an average compressive modulus of 0.38 MPa, suitable for tissue engineering applications. The printability of the bioink was further confirmed through the evaluations of morphology and diffusion rates, confirming structural integrity. Biocompatibility assessments demonstrated a high cell viability rate of 82.65% following 48 h of incubation, supporting the bioink’s suitability for facilitating cell survival. This study introduced a reliable technique for producing tissue-engineered scaffolds that exhibit outstanding mechanical characteristics and cell viability, highlighting the promise of PEGDA–SA hydrogels in bioprinting applications.

Exploring Genetic Variability and Diversity in Exotic Germplasm Collection of Cocoa (Theobroma cacao L.)

The current research investigates the genetic variation and diversity in exotic accessions of cocoa to identify materials with desired characteristics as well as important traits contributing to yield and variability. The experiment was conducted at Cocoa Research Centre, Kerala Agricultural University, from 2021 to 2023. The study comprises 23 exotic genotypes in which observations were recorded on quantitative traits on flowers, pods, and beans. Statistical analysis was carried out based on the observations recorded. The morphological evaluation revealed ample variability among the genotypes for all the quantitative characters studied. The average dry bean weight recorded was 1.08 grams, which is a desirable trait in selecting individuals with high processing efficiency for the chocolate industry. Characters such as single bean dry weight, pod value, pod index, and efficiency index have recorded high values of GCV, heritability and genetic advance as percent of the mean. Principal component (PC) analysis showed that the first three PCs with more than one Eigen-value contributed to 92.18 percent of total variability among the germplasm. Yield components viz. pod value, total bean weight pod-1 and single bean dry weight significantly contribute to maximum variability among the exotic germplasm. Moreover, genotypes were grouped into five clusters based on hierarchical agglomerative clustering technique. Hybridization program involving superior genotypes identified in diverse clusters as parents can produce high-yielding hybrids. The high values recorded for the genetic parameters by yield components suggest the selection of genotypes based on which can result in better-performing progeny.

The effect of myo-inositol antioxidant activity on human sperm parameters and DNA damage in ultra-rapid and conventional freezing methods

Male fertility preservation is still challenged by cell damage induced during sperm cryopreservation and impaired sperm structure and function. Sperm ultra-rapid freezing, despite a higher protective effect compared to conventional freezing method, is still associated with suboptimal sperm cryosurvival and needs to be modified to increase its efficiency in sperm protection. Sperm freezing media supplemented with antioxidants can improve sperm parameters following freezing-warming process. In this study, we aimed to investigate the effect of employing ultra-rapid freezing and myo-inositol on sperm cryosurvival. Thirty semen samples with normal sperm parameters were collected and each one was divided into four portions to cryopreserve by conventional freezing, ultra-rapid freezing, conventional freezing + myo-inositol 2 mg/ml, and ultra-rapid freezing + myo-inositol 2 mg/ml. Sperm samples warmed after at least 24 h of freezing and sperm cryosurvival were analyzed by evaluation of sperm motility, viability, morphology and DNA fragmentation index (DFI). Freezing method had a significant influence on post-thaw sperm DFI and morphology (p < 0.05) and the interaction between freezing method and antioxidant supplementation significantly affected sperm morphology (p < 0.05). The highest percentage of sperm normal morphology and minimal DFI was achieved using ultra-rapid freezing supplemented by myo-inositol antioxidant compared to other groups (P < 0.05). The highest sperm DNA damage after freezing-warming was observed following the conventional freezing method. In conclusion, sperm freezing method was identified as factor strongly influencing sperm DFI and morphology after thawing/warming. Sperm samples can be rapidly frozen using the modified freezing media supplemented by myo-inositol without impacting sperm DNA and morphology.

Evaluation of anterior teeth crown-root morphology and alveolar bone structure in patients with closed deep overbite using cone beam computed tomography

This retrospective study used cone-beam computed tomography to investigated the crown-root morphology and alveolar bone structure in incisors region in subjects with closed deep overbite and comparison the difference between gender and age. The CBCT images for 40 subjects (group C) with Angle II division 2 were selected from patients of the HeFei Stomatological Hospital from November 2023 to March 2024.20 individual normal occlusion subjects (group A),20 patients with Angle II division 1(group B) were included. The crown-root ratio of the maxillary and mandibular anterior teeth, crown-root angle and alveolar bone structure were measured on the CBCT images, the data were statistically analyzed. In addition to mandibular lateral incisor, the crown-root ratio of maxillary and mandibular anterior teeth in the Class II division 2 group was higher than that in the individual normal occlusion group and the Class II division 1 (P < 0. 05). The crown-root angle of maxillary anterior teeth in the Class II division 2 group was smaller than that in the individual normal occlusion group and the Class II division 1 (P < 0. 05). The alveolar bone thickness of the maxillary central incisor was comparatively smaller, while the alveolar bone height was relatively higher in the Class II division 2 group. Age and gender were associated with change in root lengths and crown-root angle for the Class II division 2 group (P < 0. 05). Patients with closed deep overbite malocclusion exhibit a significant difference compared to the controls for most measurements. The patients presenting with Class II division 2 malocclusion exhibit excessive inward positioning of the anterior teeth, resulting in evident crown-root angle, a large crown-root ratio, and minimal labial alveolar bone. In order to keep the tooth movement within the safe range of alveolar bone movement, it is necessary to strictly control the torque and use appropriate orthodontic force to reduce the risk of bone fenestration, bone dehiscence, gingival recession and root resorption.

Engineering Budesonide Microparticles for Enhancing Pulmonary Targeting: Validation through In Vivo Investigations

AbstractThe study aimed to develop and evaluate budesonide inhalation micro‐suspension to target inflammation in the lower trachea and bronchi. Initially, the slurry phase is prepared by dispersing the drug (0.25 and 0.5 mg ml−1) in the Water for Injection with polysorbate 80. The slurry is mixed with the aqueous phase comprising other excipients. The mixture is further subjected to the treatment with homogenizer. The prepared formulations are subjected to various physio‐chemical analysis like description, density, pH, osmolality, viscosity, shelf life, impurities, assay, particle size, etc. Andersen cascade impactor is used to estimate the aerodynamic performance. In vivo analysis is initiated with the induction of Asthma in Sprague Dawley rats by ovalbumin challenge intranasally and the treatment is given with the aid of a nebulizer. Lastly, the biochemical estimations are carried out in blood and bronchoalveolar lavage fluid (BALF). The lungs are also subjected to morphological and histological evaluations. The physio‐chemical and analytical examination indicated a stable formulation that is compliant with the standard guidelines. The D90 value of the test formulation is ≤4.5 µm. Cascade impactor evaluation estimated almost, 2–10% higher amount of budesonide reaching the target region as compared to the marketed product. Test formulation attenuated the manifestation of ovalbumin‐induced asthma through suppression of inflammatory cell count in blood and BALF, and modulation of Lactate dehydrogenase and Alkaline Phosphatase levels which is further confirmed by histopathological analysis. The formulated budesonide micro‐suspension demonstrated stability, safety, and efficient delivery to the target site.

Particle Size and Morphological Evaluation of Airborne Urban Dust Particles by Scanning Electron Microscopy and Bidimensional Empirical Mode Analysis

Airborne particles affect the health of the population. As particles decrease in size, they can penetrate deeper into the respiratory system, reaching the terminal bronchioles and alveoli. Particles as small as 0.1 µm in diameter may translocate into the bloodstream, potentially impacting various organs. Additionally, the smaller the particle size, the longer they remain suspended in the air, thereby increasing their deleterious damages. The aim of this work is to study the size distribution of airborne particles emitted from anthropogenic sources of air pollution, with a special emphasis on estimating the distribution of micro and nanoparticles considered the most harmful to health. The Bidimensional Empirical Mode Decomposition (BEMD) algorithm was used on micrographs of the particles obtained by Scanning Electron Microscopy (SEM). BEMD is a current empirical computational tool applied to image analysis that allows extracting non-linear heterogeneous oscillations of brightness. We studied ROFA (Residual Oil Fly Ash) from industrial sources and DEP (Diesel Exhaust Particles) from vehicular emissions as airborne particles. After collecting the particles on filters, micrographs were taken using SEM at different magnifications to which the BEMD algorithm was applied. Particle size and asymmetry distributions were obtained for each mode, allowing the identification of the most deleterious particles. The methodology employed herein is relatively simple and effective for inferring the impact of airborne particulate matter on health and the environment.