Duration Of Incubation Research Articles

Different narcotic gases and concentrations for immobilization of ostrich embryos for in-ovo imaging

In-ovo imaging using avian eggs has been described as a potential alternative to animal testing using rodents. However, imaging studies are hampered by embryonal motion producing artifacts. This study aims at systematically comparing isoflurane, desflurane and sevoflurane in three different concentrations in ostrich embryos. Biomagnetic signals of ostrich embryos were recorded analyzing cardiac action and motion. Ten groups comprising eight ostrich embryos each were investigated: Control, isoflurane (2%, 4%, and 6%), desflurane (6%, 12%, and 18%) and sevoflurane (3%, 5%, and 8%). Each ostrich egg was exposed to the same narcotic gas and concentration on development day (DD) 31 and 34. Narcotic gas exposure was upheld for 90 min and embryos were monitored for additional 75 min. Toxicity was evaluated by verifying embryo viability 24 h after the experiments. Initial heart rate of mean 148 beats/min (DD 31) and 136 beats/min (DD 34) decreased over time by 44–48 beats/minute. No significant differences were observed between groups. All narcotic gases led to distinct movement reduction after mean 8 min. Embryos exposed to desflurane 6% showed residual movements. Isoflurane 6% and sevoflurane 8% produced motion-free time intervals of mean 70 min after discontinuation of narcotic gas exposure. Only one embryo death occurred after narcotic gas exposure with desflurane 6%. This study shows that isoflurane, desflurane and sevoflurane are suitable for ostrich embryo immobilization, which is a prerequisite for motion-artifact free imaging. Application of isoflurane 6% and sevoflurane 8% is a) safe as no embryonal deaths occurred after exposure and b) effective as immobilization was observed for approx. 70 min after the end of narcotic gas exposure. These results should be interpreted with caution regarding transferability to other avian species as differences in embryo size and incubation duration exist.

Exploring the physicochemical traits, antifungal capabilities, and 3D spatial complexity of hydroxyapatite with Ag+[sbnd]Mg2+ substitution in the biocomposite thin films

The silver/magnesium doped hydroxyapatite (AgMgHAp, Ca10−x-yAgxMgy(PO4)6(OH)2, xAg=0.05 and yMg=0.02) nanocomposites coatings were deposited on Si substrate using the dip coating technique. The resulting coatings were characterized by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), Fourier transform infrared (FTIR-ATR) spectroscopy, atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS). The EDS analysis highlighted the presence of the constitutive elements of the silver/magnesium doped hydroxyapatite (AgMgHAp) nanocomposites coatings. The surface microtexture of the AgMgHAp was assessed by atomic force microscopy (AFM) technique. The AFM data suggested the obtaining of a uniform deposited layer comprised of equally distributed nanoconglomerates. FT-IR studies highlighted the presence of vibrational modes associated with the phosphate and hydroxyl groups. No bands associated with silver or magnesium were observed. The XPS analysis highlighted the presence of the constituent elements of hydroxyapatite (Ca 2p, P 2 s, O 1 s), as well as dopants (Ag 3d, Mg 1 s and Mg 2p). The antifungal evaluation of AgMgHAp coatings was carried out using the Candida albicans ATCC 10231 fungal strain. The results of the antifungal assay revealed that the AgMgHAp coatings exhibited a strong inhibitory antifungal activity. Furthermore, the data highlighted that the AgMgHAp inhibited the development of biofilm on their surface. The results revealed that the antifungal activity of the coating varied based on the duration of incubation. On the other hand, the data also showed that AgMgHAp nanocomposites coatings inhibited the fungal cell adhesion and development from the early stages of the incubation. In addition to morphological analysis, we additionally take advantage of AFM images to investigate and explore the domain of fractal and multifractal analysis applied to the films under evaluation. Our studies indicates that nanocomposite coatings made from AgMgHAp demonstrate strong antifungal properties. Our studies indicates that nanocomposite coatings made from AgMgHAp demonstrate strong antifungal properties. These results suggest the potential of AgMgHAp nanocomposite coatings as a promising solution for developing innovative antifungal devices in biomedical applications.

Identification of Elsinoë phaseoli causing bean scab in Kenya and evaluation of sporulation using five adapted techniques

AbstractThis research addresses the presence of Elsinoë phaseoli in Kenya, where information on the biology of this pathogen remains scarce. Employing a multifaceted approach, the study demonstrates the steps taken to isolate, identify, and characterize E. phaseoli as the pathogen responsible for scab on common bean. Field observations confirmed scab symptoms, particularly the prominent pod lesions. Elsinoë phaseoli was isolated from common bean using a targeted streaking method on older acervulus‐bearing lesions. Morphological examinations revealed a notable diversity within E. phaseoli colonies, consistent with the characteristics of the genus. Molecular identification through ITS‐rDNA sequencing confirmed isolate AscoSK1 obtained in this study as belonging to E. phaseoli, offering a robust species differentiation method. Assessing conidium production required the implementation of five different culture methods. An adaptation of the Scheper et al. (2013) method yielded the highest quantity of conidia from 25 colonies spaced at 1 cm apart, with a conidial yield of 5.0 × 106 conidia per 9‐cm‐diameter Petri dish. A higher conidial yield was attained after the colonies were pre‐incubated on potato dextrose agar in the dark at room temperature for 28 days, followed by a transfer to corn meal agar for an additional 2 days at 20°C. This emphasizes the pivotal influence of incubation duration and pre‐culture conditions on the process. This research provides insights into the biology of E. phaseoli and introduces an improved method for enhancing in vitro sporulation of the pathogen, setting groundwork for future research and handling.

LIFE HISTORY PARAMETERS OF MELON FRUIT FLY, BACTROCERA CUCURBITAE (COQUILLETT) (DIPTERA: TEPHRITIDAE) ON FOUR ECONOMICALLY IMPORTANT VEGETABLES

The majority of plant species that are used for consumption and medicinal purposes belongs to Cucurbitaceae. These vegetables are attacked by several insect pests including Tephritid flies. Keeping in view, four economically important cucurbit vegetables such as sponge gourd; Luffa aegyptiaca, bitter gourd; Momordica charantia, musk melon; Cucumis melo, and bottle gourd; Lagenaria siceraria were selected to investigate the life-history parameters of Bactrocera cucurbitae (Coq.) (Diptera: Tephritidae) under laboratory conditions. For this purpose, B. cucurbitae was reared in the laboratory on each host vegetable and egg hatchability %, egg incubation period, larval survival (%), duration (days), length and width of larvae, pupal parameters, male-female ratio, and age-specific fecundity were recorded. The results showedno statistical difference in hatchability percentages, larval duration, and pupal survival (%) in all tested vegetables. However, greater length and width of larvae were recorded when the B. cucurbitae larvae were provided with bottle gourd. Moreover, maximum pupal recovery was recorded in bottle and sponge gourd among all the tested vegetables. Maximum females were also recorded in bottle gourd. Adult longevity and fecundity were significantly affected by the host. Maximum female longevity (52 days) was recorded when B. cucurbitae were provided with bottle gourd and minimum (41 days) on sponge gourd. Similarly, maximum fecundity (823 eggs) was also recorded when maggots were fed on bottle gourd and the lowest (611 eggs) on a sponge gourd. In conclusion, Bottle gourd is the most preferable and nutritious host among all the tested vegetables and needs effective management strategies for B. cucurbitae. The output of the present studies would be helpful for the development of an effective integrated pest management (IPM) program for B. cucurbitae in a variety of vegetables.

Incubation and Larval Development Durations of Sterlet (A. ruthenus LINNAEUS 1758) in River Water Rearing under Near-Natural Conditions

Day-degrees [°D], also known as accumulated thermal units (ATUs), have been used to predict the duration of early development for fish. For most sturgeon species, the available data in the literature were determined under temperature-constant conditions. However, there is a lack of information on ATUs of early development for sturgeons under natural or near-natural temperature conditions. The aim of this study was to observe the duration of incubation and the duration from hatch until feeding of sterlets (Acipenser ruthenus) under near-natural conditions. This study was embedded within the LIFE-Sterlet and LIFE-Boat 4 Sturgeon project and data were gathered from the year 2017 to 2023. The rearing of fish larvae took place in the project hatchery container with Danube water without biological, chemical, or thermal water treatment to simulate natural conditions. Temperatures [°C] were monitored on a daily basis and day-degrees were calculated by summing the temperature over time. Results indicated slower larval development than described in the literature. Hatching started earliest after 7 days at 106 °D and latest after 151 °D. Feeding started between 155 and 271 °D. The findings of this study provide valuable insights into the temperature-dependent development of sterlet larvae under near-natural conditions and can assist in the design of optimal rearing of sturgeons for conservation efforts.

Impact of Hood Steaming on Tuber Vitality of Yellow Nutsedge (Cyperus esculentus)

Cyperus esculentus is a hard-to-control, destructive perennial weed propagating and spreading through rhizomes and tubers. Currently, a combination of mechanical, cultural, and chemical measures sustained over time is required for satisfactory control of arable crops. Hood steaming is a promising thermal technique for rapid depletion of the soil tuber bank. The effect of hood steaming on C. esculentus tuber vitality was investigated using a dose–response experiment on two localities in Belgium. Steaming was performed for five durations (2, 4, 8, 16, and 32 min). Moreover, the effects of tuber burial depth (5, 15, and 25 cm), genetic C. esculentus clone (three different clones), and pedohydrological parameters (sandy and sandy, loamy soil) on hood steaming efficacy were determined. Additionally, the heat sensitivity of genetically diverse C. esculentus tubers was investigated in a laboratory incubator experiment in which incubation temperature and duration varied. To control tubers at depths of 5, 15, and 25 cm, treatment durations of 8, 16, and 32 min were required, respectively. In general, clones producing large tubers showed lower heat sensitivity. As pedohydrological parameters may affect heat transfer into the soil, they may affect steaming efficacy as well. To obtain complete control of C. esculentus tubers, soil temperature should be 50 °C or more for at least 42 min. Hood steaming is a highly suitable alternative technique for a rapid and strong depletion of the soil tuber bank in small well-delineated C. esculentus patches.

Phyco-synthesis of silver nanoparticles by environmentally safe approach and their applications

In recent years, there has been an increasing interest in the green synthesis of metallic nanoparticles, mostly because of the evident limitations associated with chemical and physical methods. Green synthesis, commonly referred to as "biogenic synthesis," is seen as an alternative approach to produce AgNPs (silver nanoparticles). The current work focuses on the use of Asterarcys sp. (microalga) for biological reduction of AgNO3 to produce AgNPs. The optimal parameters for the reduction of AgNPs were determined as molarity of 3 mM for AgNO3 and an incubation duration of 24 h at pH 9, using a 20:80 ratio of algal extract to AgNO3. The biosynthesized Ast-AgNPs were characterised using ultraviolet–visible spectroscopy (UV–Vis), zeta potential, scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDX), and high-resolution transmission electron microscopy (HR-TEM) with selected area electron diffraction (SAED) patterns. The nanoparticles exhibited their highest absorption in the UV–visible spectra at 425 nm. The X-ray diffraction (XRD) investigation indicated the presence of characteristic peaks at certain angles: 38.30° (1 1 1), 44.40° (2 0 0), 64.64° (2 2 0), and 77.59° (3 1 1) according to the JCPDS file No. 04-0783. Based on SEM and TEM, the Ast-AgNPs had an average size of 35 nm and 52 nm, respectively. The zeta potential was determined to be − 20.8 mV, indicating their stability. The highest antibacterial effectiveness is shown against Staphylococcus aureus, with a zone of inhibition of 25.66 ± 1.52 mm at 250 μL/mL conc. of Ast-AgNPs. Likewise, Ast-AgNPs significantly suppressed the growth of Fusarium sp. and Curvularia sp. by 78.22% and 85.05%, respectively, at 150 μL/mL conc. of Ast-AgNPs. In addition, the Ast-AgNPs exhibited significant photocatalytic activity in degrading methylene blue (MB), achieving an 88.59% degradation in 120 min, revealing multiple downstream applications of Ast-AgNPs.

Effect of commercial probiotics and antibiotics on the growth of Campylobacter isolated from chicken meat in Ho Chi Minh city markets

This study explores the antibiotic susceptibility of Campylobacter, a prominent foodborne pathogen, isolated in Ho Chi Minh city markets and the efficacy of commercial probiotics in inhibiting these bacteria for enhancing food safety and treating Campylobacter infections. Bacteria were isolated from chicken meat in modified coal deoxycholate cefoperazone agar (mCCD), followed by characterization as per standard procedures. Ten isolates with Gram negative, catalase positive and oxidase positive characteristics were collected. Antibiotic susceptibility is ascertained through the determination of the inhibited zone and minimum inhibitory concentration (MIC) of five distinct antibiotics against Campylobacter on Muller Hinton agar plates, culminating in a comprehensive assessment after a 24-hour incubation duration. The antibiotic susceptibility results underscore substantial diversity in Campylobacter isolates among meat samples, thereby accentuating discernible distinctions among the various antibiotic products. The research also evaluated the suitability of 5 commercial probiotic products (re-named as A, B, C, D, and E for fair assessment) by examining their impact on the growth of Campylobacter colonies. The antimicrobial effect of probiotics against Campylobacter is assessed using the agar well diffusion assay and co-culture method. We obtained consistent results from two methods, indicating no variation in Campylobacter species among meat samples but significant variations among probiotic products. The outcomes of this research provide valuable insights into the antimicrobial potential of each probiotic and antibiotic on Campylobacter, informing recommendations for food hygiene practices and underscoring the role of both probiotics and antibiotics in combating Campylobacter infections.

Decreasing the ability of HCT116 cells to escape from therapy induced senescence by increasing the duration of doxorubicin treatment

Contacts: Maria Anatolievna Zamkova zamkovam@gmail.com Introduction. Due to the toxicity of high doses of chemotherapy, low concentrations used in cancer treatment leads to the development of senescence phenotype in tumor cells, characterized by a block in the cell cycle progression and the absence of division; changes in the transcriptional and metabolic profile of cells. A negative consequence of this stage is acquisition of individual cells the ability to escape from senescence and return to re-proliferation.Aim. To estimate the effect of the duration of drug treatment of HCT116 tumor cells on their ability to escape from therapy induced senescence.Materials and methods. The senescence phenotype was confirmed by the analysis of β-galactosidase activity; cell cycle analysis; estimation of protein levels by western blotting. Colonies were stained with crystal violet dye.Results. In our study, we showed that the duration of HCT116 cells incubation with low-dose doxorubicin affects their ability to return to re-proliferation – increasing the treatment time using same drug dose reduces the process of colony formation. The duration of doxorubicin treatment does not affect the formation of the senescence phenotype, which was confirmed by analyzing different markers of this stage (changes in β-galactosidase activity, cell cycle analysis, assessment of p21 and γH2AX protein levels). However, there is a delay in the development of cellular response to DNA damage caused by doxorubicin in cells exposed to prolong treatment protocol (increase in β-galactosidase activity, formation of polyploid cells).Conclusion. The duration of doxorubicin treatment of HCT116 cancer cells affects long-term consequences, reducing the ability of senescent cells to escape this stage when the incubation time with the drug is extended.

Soil incubation methods lead to large differences in inferred methane production temperature sensitivity

Quantifying the temperature sensitivity of methane (CH4) production is crucial for predicting how wetland ecosystems will respond to climate warming. Typically, the temperature sensitivity (often quantified as a Q10 value) is derived from laboratory incubation studies and then used in biogeochemical models. However, studies report wide variation in incubation-inferred Q10 values, with a large portion of this variation remaining unexplained. Here we applied observations in a thawing permafrost peatland (Stordalen Mire) and a well-tested process-rich model (ecosys) to interpret incubation observations and investigate controls on inferred CH4 production temperature sensitivity. We developed a field-storage-incubation modeling approach to mimic the full incubation sequence, including field sampling at a particular time in the growing season, refrigerated storage, and laboratory incubation, followed by model evaluation. We found that CH4 production rates during incubation are regulated by substrate availability and active microbial biomass of key microbial functional groups, which are affected by soil storage duration and temperature. Seasonal variation in substrate availability and active microbial biomass of key microbial functional groups led to strong time-of-sampling impacts on CH4 production. CH4 production is higher with less perturbation post-sampling, i.e. shorter storage duration and lower storage temperature. We found a wide range of inferred Q10 values (1.2–3.5), which we attribute to incubation temperatures, incubation duration, storage duration, and sampling time. We also show that Q10 values of CH4 production are controlled by interacting biological, biochemical, and physical processes, which cause the inferred Q10 values to differ substantially from those of the component processes. Terrestrial ecosystem models that use a constant Q10 value to represent temperature responses may therefore predict biased soil carbon cycling under future climate scenarios.

The Cytotoxic Effect of Ciprofloxacin Laetrile Combination on Esophageal Cancer Cell Line.

aim of this study was to examine the synergistic effect between the antibacterial drug ciprofloxacin and the natural compound laetrile on esophageal cancer cells, specifically focusing on their combined cytotoxic effect. The combined cytotoxic effects of two alternative incubation durations (24 and 72 hours) were studied using an esophageal cancer cell line. Ciprofloxacin, laetrile, and their combinations were tested at concentrations ranging from 1 to 1000 micrograms/milliliter, to enhance the safety of the combination, the concentrations of the combination constituents were reduced by half compared to when they are used individually, the combination index was then calculated to estimate the components' possible synergistic effects. The results indicate that the combined cytotoxicity of ciprofloxacin and laetrile was greater than the cytotoxicity of either ciprofloxacin or laetrile alone, the combination cytotoxicity increased with higher concentrations and longer incubation periods, in other words, the cytotoxicity pattern of the combination was time-dependent (cell-cycle specific), and concentration dependent, (cell-cycle non-specific). The study found that the combination of ciprofloxacin and laetrile had a greater inhibitory effect on the growth of esophageal cancer cells compared to ciprofloxacin or laetrile alone. This suggests a synergistic effect between the components of the mixture, which can be attributed to a complementary mechanism between the ingredients in the combination.

The effects of green and chemically-synthesized copper oxide nanoparticles on the production and gene expression of morphinan alkaloids in Oriental poppy

Oriental poppy (Papaver orientale L.) belonging to the Papaveraceae family, has the capacity to synthesize a wide range of benzylisoquinoline alkaloids (BIAs). This experiment was conducted to investigate the effects of green and chemical copper oxide nanoparticles (CuO NPs) elicitors on oxidative stress and the BIAs biosynthesis pathway in the cell suspension culture of P. orientale. This research shows that both green and chemical CuO NPs at concentrations of 20 mg/L and 40 mg/L, induce oxidative stress in the cell suspension of P. orientale by increasing the production of H2O2 and the activity of antioxidant enzymes. The comparison of treatments revealed that utilizing a lower concentration of CuO NPs (20 mg/L) and extending the duration of cell suspension incubation (up to 48 h) play a more influential role in inducing the expression of the BIAs biosynthesis pathway genes (PsWRKY, TYDC, SalSyn, SalR, SalAT, T6ODM, COR and CODM) and increasing the production of morphinan alkaloids (thebaine, codeine, and morphine). The overarching results indicate that the concentration of CuO NPs and the duration of cell treatment have a more significant impact than the nature of CuO NPs in inducing oxidative stress and stimulating the expression of the BIAs pathway genes.

CHARACTERISTICS FOR SOME RABBIT HEMORRHAGIC DISEASE VIRUS (RHDV) STRAINS PROPAGATED IN ISOLATED RENAL CELL OF RABBIT

The renal cells were isolated from the kidney of different week-age of healthy rabbits by a single two-stage pass perfusion method and suspended in Hanks’ balanced salt solution ([Formula: see text]-, [Formula: see text]-free). The field strains of RHDV were inoculated into the suspension and incubated for 4 days at 37∘C. After incubation, the HA titer of each strain of RHDV in supernatant of virus growth was measured using human erythrocyte O type. There were differences in the HA titers of RHDV strains according to the renal cell of different week-age of rabbits, the number of hepatocyte, the strain of virus (RHDV-1, RHDV-2, RHDV-3) and the duration of incubation. The HA titer was highest when the virus was cultured in the renal cells isolated from more than 12 week-aged rabbits. And the HA titer of RHDV propagated in isolated renal cell was higher than hepatocytes. When immunized rabbits by inactivated virus strains (RHDV-1, RHDV-2 and RHDV-3), two strains, RHDV-1 and RHDV-3, showed the same level of HI titers with inactivated homogenate, but RHDV-2 did lower level of titer than the homogenate.

Biodegradation of sugarcane trash into substrates of utilizable products using lignocellulolytic microorganisms

Biodegradation of sugarcane trash was conducted using lignocellulolytic microorganisms, Bacillus subtilis (MW715021) and Schizophyllum commune (MW720154) strains. Throughout the biodegradation process, various physicochemical, biochemical, and microbial parameters were monitored. The weight, organic carbon, and C:N ratio of the sugarcane residue progressively decreased from 20 to 120 days of incubation. Among different treatments, T7 (Sugarcane trash + Bacillus subtilis + Schizophyllum commune) exhibited significantly the lowest weight, organic carbon, and C:N ratio of 61.60 g, 17.85%, and 13.02, respectively. The pH of the substrate decreased slightly at the initial period of biodegradation then it increased slightly and became stable at near about neutral pH. The Electrical Conductivity (EC) of the substrate was increased with the advancement of incubation periods. The Total Kjeldahl Nitrogen (TKN) content of the substrate increased with the duration of incubation. A higher TKN content of 1.37% was recorded in T7 on 120 days of incubation. The nutrient content (P2O5, K2O, Fe, Mn, Zn, and Cu) of the substrate was increased with the advancement of the decomposition period. The cellulose and lignin content was reduced from 40.78 to 21.54 % to 23.16 and 10.84 %, respectively in T7. Lignocellulolytic enzymes i.e. cellulase, laccase, lignin peroxidase, and manganese peroxidase were increased at initial degradation periods up to 60 days then it declined at a later stage of biodegradation. The microbial population increased with incubation periods up to 60 days then after it was declined at a later stage of biodegradation.

In Vitro Cytotoxicity of Linseed Oil as Bio-Based Plasticiser in Polymethylmethacrylate (PMMA) Denture Base Material

Objectives: Polymethyl methacrylate (PMMA) is a polymer that is vastly utilised as denture base material. Previously synthetic plasticiser like Dibutyl phthalate was previously used in the PMMA. However, there are reports of ester leaching causing intraoral toxicity. Thus, bringing the awareness about organic plasticiser usage, like linseed oil. This study aims to evaluate the cytotoxicity of linseed oil to ensure its safety to be used as the natural-based plasticiser in PMMA as denture base material. Materials and Methods: The cytotoxicity of linseed oil was evaluated by culturing Human Gingival Fibroblast cells with 1, 3, 5, 7, and 10% linseed oil for 24, 48, and 72 hours. The WST-1 assay was then used to evaluate cell viability, with cell quantification performed using a spectrophotometer at a wavelength range of 420-480nm. An unpaired t-test was used to analyse all parameters' quantitative cell viability data. Results: The WST-1 Assay demonstrated that linseed oil exhibited greater than 95% cell viability at all doses and incubation durations, indicating biocompatibility. At 72 hours of incubation, there are statistically significant differences between the cells incubated in the Linseed Oil as compared to Dimethyl Sulfoxide (DMSO), which acts as a positive control. Conclusions: The study proves that linseed oil is non-cytotoxic with the Human Gingival Fibroblast cells and safe to use when incorporated later in the PMMA as denture base material.

Systematic development of immunohistochemistry protocol for large cryosections-specific to non-perfused fetal brain

BackgroundImmunohistochemistry (IHC) is an important technique in understanding the expression of neurochemical molecules in the developing human brain. Despite its routine application in the research and clinical setup, the IHC protocol specific for soft fragile fetal brains that are fixed using the non-perfusion method is still limited in studying the whole brain. New methodThis study shows that the IHC protocols, using a chromogenic detection system, used in animals and adult humans are not optimal in the fetal brains. We have optimized key steps from Antigen retrieval (AR) to chromogen visualization for formalin-fixed whole-brain cryosections (20 µm) mounted on glass slides. ResultsWe show the results from six validated, commonly used antibodies to study the fetal brain. We achieved optimal antigen retrieval with 0.1 M Boric Acid, pH 9.0 at 70°C for 20 minutes. We also present the optimal incubation duration and temperature for protein blocking and the primary antibody that results in specific antigen labeling with minimal tissue damage. Comparison with existing methodsThe IHC protocol commonly used for adult human and animal brains results in significant tissue damage in the fetal brains with little or suboptimal antigen expression. Our new method with important modifications including the temperature, duration, and choice of the alkaline buffer for AR addresses these pitfalls and provides high-quality results. ConclusionThe optimized IHC protocol for the developing human brain (13–22 GW) provides a high-quality, repeatable, and reliable method for studying chemoarchitecture in neurotypical and pathological conditions across different gestational ages.

Synthesis of bioactive glass-ceramics: Effect of the incubation duration on mechanical properties using PBS solution

Owing to its wear resistance and mechanical performance, glass-ceramics are often favoured in dental applications. However, prolonged immersion can cause material degradation and reduced strength. Typically, adding fluoride is believed to improve their properties. Hence, this work prepared fluoride-free and fluoride glass-ceramics via the melt-quenching technique. The samples were assessed after soaking PBS solution for 14 days. The fluoride sample exhibited a higher hardness of HV = 653 ± 2.80 and toughness of 3.37 ± 0.01 MPa·m1/2. This experimental results were compared to the human dentine (HV = 61.2), human enamel (HV = 510) and commercial dental glass-ceramics (HV = 408–663). The fluoride samples showed higher bioactivity, evident from the crystal formation in micrographs. Therefore, the prepared samples exhibit a promising potential as new materials for dental crown applications.

Cyclizine induces cytotoxicity and apoptosis in macrophages through the extrinsic and intrinsic apoptotic pathways.

Cyclizine, an over-the-counter and prescription antihistamine, finds widespread application in the prevention and treatment of motion sickness, encompassing symptoms such as nausea, vomiting, dizziness, along with its effectiveness in managing vertigo. However, the overuse or misuse of cyclizine may lead to hallucinations, confusion, tachycardia, and hypertension. The molecular mechanisms underlying cyclizine-induced cytotoxicity and apoptosis remain unclear. During the 24 h incubation duration, RAW264.7 macrophages were exposed to different concentrations of cyclizine. Cytotoxicity was assessed through the lactate dehydrogenase assay. Flow cytometry employing annexin V-fluorescein isothiocyanate and propidium iodide was utilized to evaluate apoptosis and necrosis. Caspase activity and mitochondrial dysfunction were evaluated through a fluorogenic substrate assay and JC-1 dye, respectively. Flow cytometry employing fluorogenic antibodies was utilized to evaluate the release of cytochrome c and expression of death receptor, including tumor necrosis factor-α receptor and Fas receptor. Western blotting was utilized to evaluate the expression of the Bcl2 and Bad apoptotic regulatory proteins. The findings unveiled from the present study demonstrated that cyclizine exerted a concentration-dependent effect on RAW264.7 macrophages, leading to the induction of cytotoxicity, apoptosis, and necrosis. This compound further activated the intrinsic apoptotic pathway by inducing mitochondrial dysfunction, Bcl2/Bad exchange expression, cytochrome c liberation, and activation of caspases contained caspase 3, 8, and 9. Moreover, the activation of the extrinsic apoptotic pathway was observed as cyclizine induced the upregulation of death receptors and increased caspase activities. Based on our investigations, it can be inferred that cyclizine prompts cytotoxicity and apoptosis in RAW264.7 macrophages in a concentration-dependent manner by triggering both the intrinsic and extrinsic apoptotic pathways.

THE INFLUENCE OF PH ON THE ANTIBACTERIAL ACTIVITY OF THERMOPHILIC BACTERIA ISOLATED FROM A MOUNTAIN CRATER IN WEST JAVA AGAINST STREPTOCOCCUS MUTANS

Thermophilic bacteria that live in hot springs are a potential source of valuable biochemical capable of producing antibacterial regarding its ability to produce antimicrobial peptide (AMP). The study aims to investigate how pH affects the antibacterial activity of a bacterium isolated from a mountain crater in West Java against Streptococcus mutans, the predominant bacterium in the creation of oral biofilms. According to Buchanan and Gibbons, samples collected from the water of Mount TangkubanPerahus crater were identified. The ideal pH and incubation duration were ascertained using a disc diffusion antibacterial test against S. mutansATCC 25175, considering the diameter of the inhibitory zone. The sample was determined to be a Gram-positive bacterium that, at its ideal pH, forms the highest inhibitory zone, indicating antibacterial activity against S. mutans. At an optimum pH of 6-8, thermophilic bacteria isolated from a mountain crater in West Java have antibacterial activity against S. mutans.

The Application of Cows Rumen for Electricity Generation Through The Implementation of A Ceramic-Based Microbial Fuel Cell System

The growth in population is not adequately matched by the corresponding increase in energy demand. The imperative for prioritizing the exploration of alternative energy sources that possess attributes of safety, affordability, and ample access to raw materials cannot be overstated. An energy source with significant potential is a microbial fuel cell (MFC)-based energy source. This study aims to investigate the utilization of cow rumen as a substrate and source of nutrition in the bioenergy system of microbial fuel cells (MFCs). The present study aims to investigate the impact of substrate concentration and bacterial incubation duration derived from bovine rumen bacteria on the attainment of optimal power density. The research employed a Dual Chamber Microbial Fuel Cell (MFC) device using a Proton Exchange Membrane (PEM) constructed from ceramic materials. Daily observations were conducted over a period of 30 days. The performance of MFC was assessed utilizing the polarization technique. The findings indicated that altering the proportion of bovine rumen bacterial substrates and the duration of bacterial incubation had an impact on the power density seen in the Microbial Fuel Cell (MFC) system. The optimal conditions were attained when the substrate concentration reached 3640 ppm and after 7 days, resulting in a maximum power density of 864 mW/m2.