Higher Cell Numbers Research Articles

Evaluation of the CellaVision® DM-1200 system for detecting and quantifying schistocytes

Detection of schistocytes is an important first step in the differential diagnosis of thrombotic microangiopathy. It is however labor intensive and prone to subjectivity. To improve and standardize the detection and quantification of schistocytes, we studied its automated analysis by digital microscopy DM1200 (CellaVision®) on 63 positive and 102 negative smears obtained from SP-50 (Sysmex®). Easy to use and very useful for staff training, it showed a lower between-observer coefficient of variation than usually described for manual counting (25% vs. 50%). Very sensitive (100%) in pre-classification, the detection of schistocytes was highly sensitive (98.4%) and specific (96.8%) after reclassification (AUCROC =0.9929), showing a good correlation with manual microscopy. TAT was comparable to manual counting. For positive smears, the percentage of schistocytes was similar between pre- and post-classification. However, 29.6% of pre-classified schistocytes were removed and 21.8% were added. For negative smears a significative overestimation of schistocytes (292%) was observed. Except poikilocytosis, on negative smears, the most common error of the software (24.9%) was due to platelets classified in schistocytes. Were also observed for example erroneous divisions of the image (3.2%) or artifactual schistocytes resulting from scratches in the smear (2.6%). Another limit is the high number of red blood cells not analyzed (46.8% for high-density smears), which might false the schistocytes percentage. To conclude, CellaVision® technology showed many benefits, but also limits that the operator needs to know.

Low-Power Radiation-Hardened Static Random Access Memory with Enhanced Read Stability for Space Applications

In space environments, radiation particles affect the stored values of SRAM cells, and these effects, such as single-event upsets (SEUs) and single-event multiple-node upsets (SEMNUs), pose a threat to the reliability of systems used in the space industry. To mitigate the impacts of SEUs and SEMNUs, this paper proposes the Read Stability Improved and Low Power (RSLP16T) SRAM cell. It was confirmed that in SEU-induced simulations, all nodes of the RSLP16T could be restored with a charge amount of less than 100 fC. Additionally, it was verified that a similar level of restoration was possible for SEMNUs occurring in pair of storage nodes. The proposed cell achieves a high level of read stability due to a high pull-down cell ratio (current ratio, CR) at the storage nodes and the fact that only a pair of nodes is in contact with the bit lines during read operations. Because all node paths use a stacking structure for internal transistor configuration and a relatively higher number of cells are composed of PMOS, it consumes the least hold power. While these improvements come at the cost of slightly increased delay time and area, performance evaluation revealed that the equivalent quality metric (EQM) was the highest, indicating that the benefits outweigh the drawbacks. The proposed integrated circuit is implemented in the 90 nm CMOS process and operated on 1 V supply voltage.

Diagnostic features in paediatric MDS-EB with UBTF-internal tandem duplication: defining a unique subgroup.

Tandem-duplications of the UBTF gene (UBTF-TDs) have recently been identified as a new genetic driver in young individuals with acute myeloid leukaemia (AML) and myelodysplastic syndrome (MDS). Disease in these newly defined subgroups is characterized by poor response to standard intensive chemotherapy and inferior survival of the affected patients. However, a thorough analysis of bone marrow histomorphology of UBTF-mutated neoplasia has not been undertaken thus far. In this retrospective study, we investigated the characteristic histopathological features of a cohort comprising 14 paediatric MDS patients with an excess of blasts (MDS-EB) and UBTF-TD. Bone marrow biopsies from these patients revealed hypercellularity and severe dysplasia across all three haematopoietic lineages. In particular, a marked hyperplastic megakaryopoiesis characterized by the presence of frequent micromegakaryocytes and a high number of monolobulated cells forming small clusters was observed. Additionally, erythropoiesis was left-shifted, with numerous blastoid precursors. The granulopoietic precursors displayed prominent UBTF-positive nucleoli. The unique combination of these histomorphological features strongly suggests a possible UBTF aberration. It will allow initiating the appropriate genetic testing to confirm the presence of UBTF-TD and identify potential additional genetic alterations. Such molecular profiling will not only contribute to a better understanding of the disease mechanism, but also facilitate more rational treatment approaches for these high-risk paediatric MDS patients.

Innovative Biocompatible Blend Scaffold of Poly(hydroxybutyrate-co-hydroxyvalerate) and Poly(ε-caprolactone) for Bone Tissue Engineering: In Vitro and In Vivo Evaluation.

This study evaluated the biocompatibility of dense and porous forms of Poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV), Poly(ε-caprolactone) (PCL), and their 75/25 blend for bone tissue engineering applications. The biomaterials were characterized morphologically using scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR), and the thickness and porosity of the scaffolds were determined. Functional assessments of mesenchymal stem cells (MSCs) included the MTT assay, alkaline phosphatase (ALP) production, and morphological and cytochemical analyses. Moreover, these polymers were implanted into rats to evaluate their in vivo performance. The morphology and FTIR spectra of the scaffolds were consistent with the expected results. Porous polymers were thicker than dense polymers, and porosity was higher than 92% in all samples. The cells exhibited good viability, activity, and growth on the scaffolds. A higher number of cells was observed on dense polymers, likely due to their smaller surface area. ALP production occurred in all samples, but enzyme activity was more intense in PCL samples. The scaffolds did not interfere with the osteogenic capacity of MSCs, and mineralized nodules were present in all samples. Histological analysis revealed new bone formation in all samples, although pure PHBV exhibited lower results compared to the other blends. In vivo results indicated that dense PCL and the dense 75/25 blend were the best materials tested, with PCL tending to improve the performance of PHBV in vivo.

Impact of lipid content on oxygen reactive species and viability predictors in vitrified bovine embryos

Given the significant variation in lipid levels among bovine embryos, our study was designed to associate lipid content to oxidative stress in individual embryos undergoing vitrification, and to assess how this and other morphological parameters impacts cryosurvival. Linear and logistic regression were performed to understand the influence of the variables in the cryosurvival. T-test or Kruskal Wallis were employed to compare means. Vitrified embryos revealed a positive correlation between lipid content and oxidative stress post-warming both 2 h (p = 0.025, n = 64) and 48 h (p < 0.001, n = 122) after warming. Lipid levels explained (p < 0.001) up to 51 % (multiple R-squared) of oxidative stress variability. Compared to fresh embryos, a negative influence (p = 0.01) of vitrification-warming procedures was detected in lipid levels. Vitrified embryos exhibited lower (p < 0.001, n = 90) mean lipid content compared to fresh counterparts 48 h post-warming, and similar (p = 0.24) oxidative stress levels. No impact of lipid content or oxidative stress levels was detected on hatchability or embryo quality 48 h post-warming (n = 99). Expansion just after (0 h) and 2 h after warming resulted in a higher chance of hatching (p = 0.015 and p = 0.008, OR 1.30 and 1.58), and a positive association was observed between expansion at 0 h (p = 0.002) and embryo area (p = 0.047) with cell number. In conclusion, a decrease in lipid levels was found following vitrification-warming procedure and an individual association between lipids and oxidative stress is present in vitrified embryos. Lipids or oxidative stress levels was not linked to survivability of vitrified embryos 48 h following warming. Expansion at 0 h indicates a better chance for hatching and higher cell numbers in vitrified embryos.

Optimization of iron, phosphate, and salinity in nutrient medium using response surface methodology for enhancing biochemical composition in Chlorella sp. culture

Microalga Chlorella sp. is a highly scientifically and commercially attractive unicellular microorganism and has developed a stable industry as a nutritional supplement for people and animals. Various nutrient media have been used to grow Chlorella sp. cultures to enhance their growth, pigmentation, and lipid content. However, the optimal biochemical composition and density of Chlorella sp. cultures require an understanding of optimizing the nutrient medium to study their production. The present study aims to investigate the effects of iron, phosphate, and salinity concentrations by working synergically in a nutrient medium on the growth responses, pigments, and lipid accumulation of Chlorella sp. culture using a response surface methodology (RSM) approach. Using the result from the RSM software, a total of 18 experimental groups (E1 – E18) were evaluated, and one confirmation (C) study was conducted. The results revealed that the E7 experiment in Chlorella sp. culture provided the highest cell density and specific growth rate (SGR) with 176.00 × 106 cells mL−1 and 0.35 day−1, respectively. Similarly, the E5 and E1 experiments produced the highest cell density of 166.10 × 106 cells. mL−1 and 152.13 × 106 cells mL−1, respectively. The SGR was also increased at 0.33 day−1 in the E5 experiment and 0.33 day−1 in the E1 experiment. Consequently, the culture of Chlorella sp. containing high iron and phosphate concentrations and lower salinity in a nutrient medium had the highest number of cells, SGR, and pigment accumulation (chlorophyll a and total carotenoid). In addition, the presence of high salinity concentrations reduces Chlorella sp. growth. However, the increase in the growth of Chlorella sp. culture did not indicate an increase in other biochemical compositions. In some cases, biochemical compositions are high due to nutritional limitations or stress factors. For example, in pigment accumulation, chlorophyll a pigment accumulation was increased in experiment E7 (51.57 μg mL−1), while total carotenoid accumulation was increased in experiment E8 (20.68 μg mL−1). In addition, increasing salinity concentration increased chlorophyll a and total carotenoid contents per cell, but decreased Chlorella sp. growth as shown in the E4 experiment, which achieved chlorophyll a levels of 1.38 pg cell−1 and total carotenoid levels of 0.52 pg cell−1. Furthermore, Chlorella sp. culture produces a higher lipid accumulation of 37.38 % in the E3 experiment. Hence, the results of this study contribute to understanding the optimal biochemical composition and cell growth of Chlorella sp. cultures.

Effects of silicon nitride (Si3N4) incorporation on physicochemical, bioactivity and antibacterial properties of 45S5 bioactive glass

In this study, silicon nitride (Si3N4, SN) particles (10–30 wt%) were introduced into 45S5 bioactive glass (BG), and the impact of the incorporation on thermal and physicochemical features of the composite was analyzed. SN inclusion shifted the crystallization temperature of BG to higher temperatures, thus, the viscous flow of the glassy phase enhanced the sintering-ability and eventually reduced the sintering temperature from 1050 °C to 900 °C. XRD results revealed that increased SN fractions sustained higher amorphous content while decreasing the fraction of the main BG crystallization phase (combeite). BG + SN composites were cytocompatible with MG-63 cells. A higher number of cells was visualized on the BG + SN pellets, yet no statistically significant viability difference was determined between the BG + SN and BG samples. SN addition also endowed antibacterial activity to the BG + SN composites against S. aureus and E. coli pathogens, revealing that SN addition to BG is an alternative approach to developing biocompatible, bioactive, and antibacterial materials.

Glandular cell products in adult cestode: A new tale of tapeworm interaction with fish innate immune response

The caryophyllidean tapeworm Caryophyllaeus brachycollis (Janiszewska, 1953) is indigenous to the Lake Blidinje in the west-central part of Bosnia-Herzegovina where it infects chub Squalius tenellus (Heckel, 1843). Of 22 chubs examined, 45% were infected with C. brachycollis and a total of 912 specimens of this worm were counted. Histopathological and ultrastructural investigations were conducted on interface region between chub intestine and cestode scolex. Different sizes of lipid droplets in cestode tegument, in interface region and in chub enterocytes were observed. C. brachycollis lacks any specialized attachment organs and with an expanded, flattened scolex goes deep in mucosal folds and firmly attaches to them. In the epithelium of fish intestine, near the site of worm attachment, a high number of mucous cells and several rodlet cells were noticed. Indeed, within the intestinal tunica propria-submucosa, beneath the site of scolex attachment, numerous neutrophils and mast cells were encountered. Transmission electron microscopy of the apical part of the scolex of C. brachycollis showed the occurrence of a multicellular, syncytial glandular complex, the scolex produced membrane-bound secretory granules and their fibrillar contents discharged by merocrine and apocrine secretion onto the host-parasite interface. Our results are among the first to provide evidence on the sophisticated relationship between fish intestine and amorphous-undefinable substance produced by scolex glandular complex.

COMSE: analysis of single-cell RNA-seq data using community detection-based feature selection

BackgroundSingle-cell RNA sequencing enables studying cells individually, yet high gene dimensions and low cell numbers challenge analysis. And only a subset of the genes detected are involved in the biological processes underlying cell-type specific functions.ResultIn this study, we present COMSE, an unsupervised feature selection framework using community detection to capture informative genes from scRNA-seq data. COMSE identified homogenous cell substates with high resolution, as demonstrated by distinguishing different cell cycle stages. Evaluations based on real and simulated scRNA-seq datasets showed COMSE outperformed methods even with high dropout rates in cell clustering assignment. We also demonstrate that by identifying communities of genes associated with batch effects, COMSE parses signals reflecting biological difference from noise arising due to differences in sequencing protocols, thereby enabling integrated analysis of scRNA-seq datasets of different sources.ConclusionsCOMSE provides an efficient unsupervised framework that selects highly informative genes in scRNA-seq data improving cell sub-states identification and cell clustering. It identifies gene subsets that reveal biological and technical heterogeneity, supporting applications like batch effect correction and pathway analysis. It also provides robust results for bulk RNA-seq data analysis.

From follicle to blastocyst: microRNA-34c from follicular fluid-derived extracellular vesicles modulates blastocyst quality

BackgroundWithin the follicular fluid, extracellular vesicles (EVs) guide oocyte growth through their cargo microRNAs (miRNAs). Here, we investigated the role of EVs and their cargo miRNAs by linking the miRNAs found in EVs, derived from the fluid of an individual follicle, to the ability of its oocyte to become a blastocyst (competent) or not (non-competent).MethodsBovine antral follicles were dissected, categorized as small (2–4 mm) or large (5–8 mm) and the corresponding oocytes were subjected to individual maturation, fertilization and embryo culture to the blastocyst stage. Follicular fluid was pooled in 4 groups (4 replicates) based on follicle size and competence of the corresponding oocyte to produce a blastocyst. Follicular fluid-derived EVs were isolated, characterized, and subjected to miRNA-sequencing (Illumina Miseq) to assess differential expression (DE) in the 4 groups. Functional validation of the effect of miR-34c on embryo development was performed by supplementation of mimics and inhibitors during in vitro maturation (IVM).ResultsWe identified 16 DE miRNAs linked to oocyte competence when follicular size was not considered. Within the large and small follicles, 46 DE miRNAs were driving blastocyst formation in each group. Comparison of EVs from competent small and large follicles revealed 90 DE miRNAs. Cell regulation, cell differentiation, cell cycle, and metabolic process regulation were the most enriched pathways targeted by the DE miRNAs from competent oocytes. We identified bta-miR-34c as the most abundant in follicular fluid containing competent oocytes. Supplementation of miR-34c mimic and inhibitor during IVM did not affect embryo development. However, blastocyst quality, as evidenced by higher cell numbers, was significantly improved following oocyte IVM in the presence of miR-34c mimics, while miR-34c inhibitors resulted in the opposite effect.ConclusionThis study demonstrates the regulatory effect of miRNAs from follicular fluid-derived EVs on oocyte competence acquisition, providing a further basis for understanding the significance of miRNAs in oocyte maturation and embryonic development. Up-regulation of miR-34c in EVs from follicular fluid containing competent oocytes and the positive impact of miR-34c mimics added during IVM on the resulting blastocysts indicate its pivotal role in oocyte competence.

Magnetic Stirring Device for Limiting the Sedimentation of Cells inside Microfluidic Devices.

In experiments considering cell handling in microchannels, cell sedimentation in the storage container is a key problem because it affects the reproducibility of the experiments. Here, a simple and low-cost cell mixing device (CMD) is presented; the device is designed to prevent the sedimentation of cells in a syringe during their injection into a microfluidic channel. The CMD is based on a slider crank device made of 3D-printed parts that, combined with a permanent magnet, actuate a stir bar placed into the syringe containing the cells. By using A549 cell lines, the device is characterized in terms of cell viability (higher than 95%) in different mixing conditions, by varying the oscillation frequency and the overall mixing time. Then, a dedicated microfluidic experiment is designed to evaluate the injection frequency of the cells within a microfluidic chip. In the presence of the CMD, a higher number of cells are injected into the microfluidic chip with respect to the static conditions (2.5 times), proving that it contrasts cell sedimentation and allows accurate cell handling. For these reasons, the CMD can be useful in microfluidic experiments involving single-cell analysis.

The effect of surface roughness and wettability on the adhesion and proliferation of Saos-2 cells seeded on 3D printed poly(3-hydroxybutyrate)/polylactide (PHB/PLA) surfaces

The aim of this work is to explore the effect of surface characteristics of the 3D printed scaffolds on their suitability for Saos-2 cells growth. Two promising materials for bone tissue engineering were examined, both containing poly(3-hydroxybutyrate)/poly(d,l-lactide) plasticized blend and one of them filled with bioactive tricalcium phosphate. The surface free energy (SFE) of samples was determined by contact angle measurement with four liquids. For non-filled sample the water contact angle (WCA) was 74° and its SFE was 40 mN m−1. The composite sample exhibited substantially decreased WCA, 61°. Moreover, the addition of the tricalcium phosphate caused doubling of the polar component of the SFE which increased by a total of 13%. 3D printed surfaces prepared by fused deposition modeling method showed a profound increase of WCA due to the increase in their surface roughness which was analyzed by confocal microscopy. The change in wetting properties strongly affected the behavior of Saos-2 cells on printed surfaces. The number of cells as measured by DNA quantification linearly decreased with increasing surface WCA. The highest number of cells was observed on the pressed samples and 3D printed surfaces made of simple lines and a grid with 50 μm gap between lines. The favored surfaces exhibit WCA under 80° which is important information for the future design of the scaffolds.

Hyperpolarized Nano-NMR Platform for Quantification of Mass Limited Samples.

Metabolic flux analysis of live cells using NMR enables the study of cancer metabolism and response to treatment. However, conventional NMR platforms require often prohibitively high numbers of cells to achieve significant resolution. In this work, we present a double 1H/13C resonance NMR probe consisting of a solenoid coil with a less than 100 nL sensitive region. In-solution robustness is demonstrated through measurement of decaying hyperpolarized signals. A suspension of live cells and hyperpolarized (HP) [1-13C]pyruvate is loaded in the coil, and dynamic changes in pyruvate and lactate concentrations by fractions of femtomoles are detected from just 2000 live cells at a time, in seconds. Through an integrated microfluidic channel, the probe is used as high-throughput platform to perform nondestructive quantitative analysis of metabolic flux of different leukemia cell lines with sensitivity to detect on target treatment response. This approach platform provides an approach to study mass-limited samples and living cells with dramatically enhanced sensitivity in real time.

Acute effects of atrazine on the immunoexpressions of sertoli and germ cells molecular markers, cytokines, chemokines, and sex hormones levels in mice testes and epididymides

Atrazine is currently one of the most commonly used agrochemicals in the United States and elsewhere. Here, we studied the immunoexpression of molecular markers of mammalian testicular functions: androgen receptor (AR), promyelocytic leukemia zinc finger (PLZF), GDNF family receptor alpha-1 (GFRA1), VASA/DDX4 (DEAD-Box Helicase 4) as well as the levels of intratesticular and intra-epididymal estradiol (E2) and dihydrotestosterone (DHT), tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukins (IL-1β and IL-6, IL-10) and testicular chemokines (CXCL-1, CCL-2 and CCL3) in BalB/c mice after a sub-acute gavage treatment with a gonado-toxin, atrazine (50 mg/kg body wt.) for three days. We found high numbers of AR immunopositive Sertoli cells and low numbers of GFRA1, PLZF and VASA/DDX4-positive germ cells in the seminiferous tubule regions of the testes. While TNF-α level in the testes fell and remained unchanged in the epididymides, IFN-γ levels in the testes remained constant but increased in the epididymides. E2 and DHT concentrations remained unaltered in the testes but were changed in the epididymides. There were no significant changes in the levels of interleukins in the testis and epididymis. Intratesticular chemokines were also not significantly altered, except for CCL-4, which was increased in the testis. Light microscopy of the epididymis showed detached epithelium and some detached cells in the lumen. It is concluded that atrazine changed the inflammatory status of the gonads and highlighted Sertoli and undifferentiated spermatogonia as important targets for atrazine's toxic effects in the testis of mice. Concerning the epididymis, atrazine altered the epididymal hormonal concentrations and promoted histopathological modifications in its parenchyma.

The Effects of Seleno-Methionine in Cadmium-Challenged Human Primary Chondrocytes.

Cadmium (Cd) is a potentially toxic element able to interfere with cellular functions and lead to disease or even death. Cd accumulation has been demonstrated in cartilage, where it can induce damage in joints. The aim of this study was to evaluate the effect of CdCl2 on primary cultures of human chondrocytes and the possible protective effect of seleno-methionine (Se-Met). Human primary articular chondrocytes were cultured and treated as follows: control groups, cells challenged with 7.5 μM and 10 μM CdCl2 alone, and cells pretreated with 10 and 20 μM Se-Met and then challenged with 7.5 μM and 10 μM CdCl2. Twenty-four hours after incubation, cell viability, histological evaluation with hematoxylin-eosin stain, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay were performed. Furthermore, reverse transcription-PCR was carried out to evaluate mRNA levels of BAX, BAK1, CASP-3, and CASP-9. After CdCl2 challenge at both doses, a reduced cell viability and an overexpression of BAX, BAK1, CASP-3, and CASP-9 genes, as well as a high number of TUNEL-positive cells, were demonstrated, all parameters becoming higher as the dose of CdCl2 was increased. The pretreatment with Se-Met lowered the expression of all considered genes, improved cell viability and morphological changes, and reduced the number of TUNEL-positive cells. It was concluded that Se-Met plays a protective role against CdCl2-induced structural and functional changes in chondrocytes in vitro, as it improved cell viability and showed a positive role in the context of the apoptotic pathways. It is therefore suggested that a translational, multifaceted approach, with plant-based diets, bioactive functional foods, nutraceuticals, micronutrients, and drugs, is possibly advisable in situations of environmental pollution caused by potentially toxic elements.

Making a healthy choice: Tactical host selection behaviour of a parasitoid wasp

Abstract Host selection behaviour plays a pivotal role in determining the success of parasitoids, especially in environments where host quality varies. In heterogeneous environments, parasitoids frequently encounter hosts that are infected. We conducted experiments to ascertain whether the ecto‐parasitoid Bracon brevicornis can differentiate between healthy caterpillars and those infected with Bacillus thuringiensis (Bt) and examined the implications of this host selection behaviour. In an olfactometer choice assay, B. brevicornis avoided Bt‐infected Spodoptera litura hosts. In a no‐choice assay, the female parasitoid displayed a lower paralysis rate on infected hosts, indicating a strong selection behaviour. Furthermore, we explored the impact of infection duration by exposing female wasps to larvae infected for 24, 48 and 72 h with Bt. Both choice and no‐choice assays demonstrated that female B. brevicornis refrains from ovipositing on infected larvae, regardless of the infection duration. Direct exposure to Bt through consumption did not affect the fitness of the tested parasitoid wasp. However, post‐24‐h Bt infection, host larvae showed an increased total hemocyte density, particularly high phagocytic cell numbers, and enhanced melanization, rendering the host larva unsuitable for parasitoid development. Taken together, our study underscores that parasitoids exhibit robust host selection behaviour by actively avoiding infected hosts. This insight is valuable for devising new pest control strategies in agriculture that safeguard beneficial parasitoids.

Enhancing heavy metal phytoremediation in landfill soil by Chrysopogon zizanioides (L.) roberty through the application of bacterial-biochar pellets

This work investigated the performance of bacterial-biochar pellets (BBP) on promoting heavy metal phytoremediation by Chrysopogon zizanioides (L.) Roberty planted in landfill soil. Micrococcus sp. MU1, an indole-3-acetic acid (IAA)-producing bacterium, was incorporated into biochar pellets (MU1-BBP). MU1-BBP contained a high number of viable bacterial cells with 90 % viability after storage at 4°C for two months. In addition, it had a greater survival capability in heavy metals-contaminated landfill soil than MU1-free cells. The application of MU1-BBP significantly promoted the growth performance and chlorophyll synthesis of C. zizanioides planted in landfill soil. Cd, Cu, Pb and Zn were mostly retained in the roots relative to the shoots of C. zizanioides. The accumulation of Cd, Cu, Pb, and Zn in the roots of C. zizanioides were significantly boosted by MU1-BBP inoculation. Plants with MU1-BBP inoculation enhanced the accumulation coefficient (AC) of all four heavy metals. The values of AC values of Cd, Cu, and Zn in C. zizanioides with MU1-BBP inoculation were greater than 1.0. However, translocation factors of all four metals were low. The findings suggest C. zizanioides may be a good heavy metal phytostabilizer. The findings support the potential of bacterial-biochar pellets to enhance the efficiency of heavy metal phytostabilization of C. zizanioides in landfill sites.

Biphasic bone substitutes coated with PLGA incorporating therapeutic ions Sr2+ and Mg2+: cytotoxicity cascade and in vivo response of immune and bone regeneration.

The incorporation of bioactive ions into biomaterials has gained significant attention as a strategy to enhance bone tissue regeneration on the molecular level. However, little knowledge exists about the effects of the addition of these ions on the immune response and especially on the most important cellular regulators, the macrophages. Thus, this study aimed to investigate the in vitro cytocompatibility and in vivo regulation of bone remodeling and material-related immune responses of a biphasic bone substitute (BBS) coated with metal ions (Sr2+/Mg2+) and PLGA, using the pure BBS as control group. Initially, two cytocompatible modified material variants were identified according to the in vitro results obtained following the DIN EN ISO 10993-5 protocol. The surface structure and ion release of both materials were characterized using SEM-EDX and ICP-OES. The materials were then implanted into Wistar rats for 10, 30, and 90days using a cranial defect model. Histopathological and histomorphometrical analyses were applied to evaluate material degradation, bone regeneration, osteoconductivity, and immune response. The findings revealed that in all study groups comparable new bone formation were found. However, during the early implantation period, the BBS_Sr2+ group exhibited significantly faster regeneration compared to the other two groups. Additionally, all materials induced comparable tissue and immune responses involving high numbers of both pro-inflammatory macrophages and multinucleated giant cells (MNGCs). In conclusion, this study delved into the repercussions of therapeutic ion doping on bone regeneration patterns and inflammatory responses, offering insights for the advancement of a new generation of biphasic calcium phosphate materials with potential clinical applicability.

Randomized split-mouth clinical trial comparing osteoblastic activity and osteogenic potential of autogenous particle harvesting during implant surgery without irrigation and with irrigation.

To compare the osteoblastic activity and osteogenic potential of autogenous particle harvesting during implant surgery using low-speed drilling without irrigation and high-speed drilling with irrigation. Thirty patients with bilateral missing teeth of 3.6 and 4.6 were randomized into two groups (Group 1: low-speed drilling without irrigation and Group 2: high-speed drilling with irrigation) and 60 single dental implants were placed. The temperature at the tip of each drill was recorded and the harvested bone was weighed; particle size and Ca and P levels were also analyzed. After osteoblast culture, cell viability, cell cycle assay, cell migration, vascular endothelial growth factor (VEGF) concentration, and mineralized nodule formation were assessed. Although the temperature of the drills was slightly higher in Group 1, no statistically significant differences were observed (p ≤ 0.05); however, the amount of harvested bone was higher (p < 0.001) and the size of the particles was higher (p = 0.019). In relation to osteoblastic activity and osteogenic potential, higher cell proliferation, higher number of cells in G2/M and S phases, higher cell migration capacity, higher VEGF concentration, and higher amount of mineralized nodule formation were observed in Group 1. Low-speed drilling without irrigation does not result in a significant increase in bone temperature compared to conventional drilling. However, a greater amount of bone is obtained; in addition, osteoblastic activity and osteogenic potential are higher with this technique, but further clinical studies are necessary.

Growth, Lipid, and Pigment Properties of Locally Isolated (Kastamonu, Türkiye) Chlorella sp.

Chlorella has become one of the most studied and produced microalgae, with Spirulina among the hundreds of species since the beginning of microalgal biotechnology. The growth performance of microalgae and the biochemical composition of the biomass may also vary significantly by strain. Therefore, it is thought that searching for new strains from aquatic environments is important in providing the most suitable microalgae for production. An isolated strain from Daday Stream was cultured in the laboratory at Kastamonu University. BG-11 was used as a medium, and CO2 from the air was used as a carbon source in the experiments. The initial cell number was arranged to 1.0×106 cells mL-1, and the highest cell number was found on the 17th day as 40.52×106 cells mL-1. Chlorophyll a and carotenoids were determined at the end of the experiment and were found as 3.48±0.08 µg mL-1 and 1.16±0.02 µg mL-1, respectively. Total lipid amount and fatty acid composition analysis were also conducted at the end of the study. According to the analyses, the lipid content of Chlorella sp. was found to be 15.37±0.00% (w/w). ∑SFA (saturated fatty acid), ∑MUFA (monounsaturated fatty acid), and ∑PUFA (polyunsaturated fatty acid) ratios were calculated to be 31.30±1.21%, 4.99±0.34% and 63.71±2.65%, respectively.