Medium Conditions Research Articles

Comparative growth response of three rubber (Hevea brasiliensis Muell. Arg) clones in somatic embryo propagation

Abstract Plants have the ability to grow with different responses depending on the clones and varieties of plants. In rubber plants, each clone has specific characteristics or traits that distinguish it from other clones, whether in terms of growth, latex production, or resistance to pests and diseases. The objective of this experiment was to assess the response of three rubber (Hevea brasiliensis Muell. Arg) clones to the same media conditions for the growth and development of somatic embryos (SE). Integuments from three rubber clones, namely PB 260, PB 330, and PB 340, were used as explants to induce embryonic callus on MH media. After four weeks post-induction, the callus was sub-cultured to expressed SE then SE was transferred to the RITA® bio-reactor for plantlets development in liquid MD media. Parameters for each steps, such as the percentage of contamination, percentage of callus response, color of callus, percentage of SE induction, number of stage of SE, and number of shoots were recorded. The results revealed that the highest percentage of viable callus was observed in the PB 260 clone at 23.20%. On SE expression, the highest being in clone PB 340 at 2.04%. On SE development the highest shoot growth was observed in the PB 260 clone, with a total of 5 shoots.

Validity and Reproducibility of Counter Electrodes for Linear Sweep Voltammetry Test in Microbial Electrolysis Cells

The electrode is a key component in a microbial electrolysis cell (MEC) that needs significant improvement for practical implementation. Accurate and reproducible analytical methods are substantial for the effective development of electrode technology. Linear sweep voltammetry (LSV) is an essential analytical method for evaluating electrode performance. In this study, inoculated carbon brush (IB), abiotic brush (AB), Pt wire (PtW), stainless steel wire (SSW), and mesh (SSM) were tested to find the most suitable counter electrode under different medium conditions. The coefficient of variation (Cv) of maximum current (Imax) was the most decisive indicator of the reproducibility test. This study shows that (i) the electrode used in operation is an appropriate counter electrode in an acetate-added condition, (ii) the anode LSV test should avoid the use of Pt wire as counter electrodes, and (iii) PtW is an appropriate counter electrode in cathode LSV in all conditions.

Mechanisms of cell size regulation in slow-growing Escherichia coli cells: discriminating models beyond the adder

Under ideal conditions, Escherichia coli cells divide after adding a fixed cell size, a strategy known as the adder. This concept applies to various microbes and is often explained as the division that occurs after a certain number of stages, associated with the accumulation of precursor proteins at a rate proportional to cell size. However, under poor media conditions, E. coli cells exhibit a different size regulation. They are smaller and follow a sizer-like division strategy where the added size is inversely proportional to the size at birth. We explore three potential causes for this deviation: degradation of the precursor protein and two models where the propensity for accumulation depends on the cell size: a nonlinear accumulation rate, and accumulation starting at a threshold size termed the commitment size. These models fit the mean trends but predict different distributions given the birth size. To quantify the precision of the models to explain the data, we used the Akaike information criterion and compared them to open datasets of slow-growing E. coli cells in different media. We found that none of the models alone can consistently explain the data. However, the degradation model better explains the division strategy when cells are larger, whereas size-related models (power-law and commitment size) account for smaller cells. Our methodology proposes a data-based method in which different mechanisms can be tested systematically.

Bioprocess Design and Evaluation of Hydrothermal Hydrolysates from Sargassum sp. for Enhancing Arthrospira platensis Growth and Protein Content

The proliferation of Sargassum biomass in various coastal areas has led to environmental and socio-economic problems. However, due to their unique composition, these biomasses offer versatile applications, prompting research into their potential in third-generation biorefineries. In this study, the hydrothermal processing of Sargassum sp. was evaluated under specific conditions at 190 °C/50 min and 150 °C/30 min. The resulting hydrolysates (liquid phase) were used as alternative culture media for cultivation. Nine treatments for the cultivation of Arthrospira platensis were assessed, varying the concentration of hydrothermal hydrolysates (HH) at 190 °C/50 min: T1 (5% v/v), T2 (10% v/v), and T3 (15% v/v). T4 (5% v/v), T5 (10% v/v), and T6 (15% v/v), maintaining the same HH conditions, and with the addition of 0.7 g/L NaNO3; and treatments T7, T8, and T9 had concentrations of 5%, 10%, and 15% of HH, respectively, at 150 °C/30 min with the addition of 0.7 g/L NaNO3, respectively. Each treatment was inoculated with 15% (v/v) of A. platensis. Growth kinetics were performed by sampling every three days for 24 days. Quantification of soluble proteins was performed for the best conditions of biomass production. The microalgae demonstrated the ability to grow under mixotrophic medium conditions and to utilize the available carbon sources in the culture medium. Treatment 4 has the highest biomass, with an Xmax (g/L) of 1.94 ± 0.06 and a protein production of 24.17 ± 0.86% (w/w). Therefore, this microalgal biomass can be used in the food matrix according to the biorefinery concept.

#289 Lipocalin-2 promotes cardiovascular calcification in chronic kidney disease by upregulating STAT3/NCOA4-mediated ferritinophagy and ferroptosis

Abstract Background and Aims Cardiovascular calcification (CVC) is a common complication of chronic kidney disease (CKD) and contributes to cardiovascular morbidity and mortality without any effective therapies. To date, the underlying mechanism of CKD-CVC has not been elucidated. Recent studies suggest that ferroptosis is a novel programmed cell death that may play a role in the process of CKD-CVC, but the mechanism is largely unclear. Ferritinophagy is an important inducer of ferroptosis that is regulated by nuclear receptor coactivator 4 (NCOA4). However, the relationship between ferritinophagy and CKD-CVC has not been explored. Lipocalin-2 (LCN2) is an iron-trafficking protein that has been associated with both osteogenesis and ferroptosis, but the exact role and molecular mechanism of LCN2 in CKD-CVC needs to be explored. Method A cross-sectional clinical study was utilized to examine the association between LCN2 expression and CKD-CVC in serum and calcified radial arteries of CKD patients. LCN2 knockout mice, their wild type littermates and mice infected with VSMCs-targeted adeno-associated virus encoding LCN2 gene or negative control gene were all fed by the high adenine and phosphate diet to induce CKD-CVC.VSMCs transfected with LCN2-overexpressing lentivirus, LCN2-shRNA lentivirus or LCN2 siRNA and recombinant LCN2 stimulated VSMCs were treated under the condition of calcifying medium to investigate the role of LCN2 in CKD-CVC in vitro. Cardiovascular calcification was evaluated by Von-kossa staining and tissue calcium content assay. VSMCs calcification were checked by the Alizarin Red Staining and cellular calcium content assay. Cellular reactive oxygen species (ROS) were measured by the DCFDA assay. Lipid peroxidation was measured by the fluorescence of the fatty acid analog C11-BODIPY581/591 probe. Intracellular labile ferrous iron levels were measured by fluorescence of Phen Green SK probe and ferro-orange probe. Transmission electron microscopy was also used to examine mitochondrial change of ferroptosis. Results The cross-sectional study identified that LCN2 levels in serum and radial arteries were significantly increased as the severity of CKD-CVC increased. High adenine and phosphate diet intake provokes renal fibrosis and cardiovascular calcification in vivo. In vitro, five-days high phosphate sodium stimulation induced calcium deposition and osteoblastic trans differentiation of VSMCs. Meanwhile, high phosphate also increased the levels of ferroptosis in VSMCs as evidenced by decreased cell viability, iron and ROS accumulation, upregulated PTGS2 and MDA levels, reduced glutathione peroxidase 4 (GPX4) levels and significantly mitochondrial damage in VSMCs. Intriguingly, calcium deposition in VSMCs could be reversed by the ferroptosis inhibitor ferrostatin-1. LCN2 expression was significantly upregulated both in the aorta of mice and calcified VSMCs. Compared with WT mice, deletion of LCN2 inhibited cardiovascular calcification whereas VSMC-targeted LCN2 overexpression aggravated CKD-CVC. Consistently, LCN2 knockdown also alleviated calcium deposition of VSMCs. Meanwhile, inhibition of ferroptosis were observed after LCN2 silencing as evidenced by decreased ferrous and lipid oxidation while increased levels of GPX4. Mechanically, we demonstrated high phosphate increased the protein level of NCOA4 and downregulated FTH1, which was reversed by depletion of LCN2. Knockdown of NCOA4 partly alleviated the ferritin reduction, ferroptosis, and calcification in VSMCs, indicating that NCOA4-mediated ferritinophagy was required for VSMCs calcification. Furthermore, we found that the recombinant LCN2 treatment significantly upregulated phosphorylated STAT3. STAT3 signaling inhibitor abolished the recombinant LCN2 induced NCOA4 upregulation and calcification in VSMCs. Conclusion Our findings indicate that the pro-calcific effect of high phosphate is due to VSMCs ferroptosis mediated by LCN2 upregulation. LCN2 silencing alleviates CKD-CVC by suppressing the STAT3/ NCOA4/FTH1 signaling mediated ferritinophagy. LCN2 could serve as a candidate therapeutic strategy for CKD-CVC.

Establishment of an efficient regeneration system of ‘ZiKui’ tea with hypocotyl as explants

Zikui (Camellia sinensis cv. Zikui) is a recently discovered cultivar of local purple tea in Guizhou, China. It is a purple leaf bud mutation material of Meitan Taicha (Camellia sinensis cv. ‘Meitan-taicha’) ‘N61’ strain, which is an important local germplasm resource in Guizhou. It is also a model plant for the study of anthocyanins, but the limited germplasm resources and the limitation of traditional reproduction hinder its application. Here, an efficient regeneration system is established by using hypocotyl as explants for the first time. Different plant growth regulators (PGRs) are evaluated during different regeneration processes including callus and root induction. According to our findings, using the optimal disinfection conditions, the seed embryo contamination rate is 17.58%. Additionally, the mortality rate is 9.69%, while the survival rate is measured as 72.73%. Moreover, the highest germination rate of 93.64% is observed under MS + 2.40 mg/L GA3 medium conditions. The optimal callus induction rate is 95.19%, while the optimal adventitious bud differentiation rate is 20.74%, Medium with 1.6 mg/L IBA achieved 68.6% rooting of the adventitious shoots. The survival rate is more than 65% after 6 days growth in the cultivated matrix. In summary, our research aims to establish a regeneration system for Zikui tea plants and design a transformation system for tea plant tissue seedlings. This will enable transfer of the target gene and ultimately facilitate the cultivation of new tea varieties with unique characteristics.

Compositional alteration of low dimensional nickel sulfides under hydrogen evolution reaction conditions in alkaline media

Developing cost-effective electrodes for electrochemical water splitting with minimal kinetic losses remains a primary challenge in water electrolysis. Nickel-based dichalcogenides, particularly sulfides, have emerged as promising candidates for the hydrogen evolution reaction (HER). However, the stability of various nickel sulfide (NixSy) phases under cathodic potentials in alkaline electrolytes is subject to debate. In this study, we investigate the HER activity and durability of both 2-dimensional (2D) and 3-dimensional (3D) NixSy nanostructures with diverse chemical compositions. Through cross-correlation analysis of spectroscopic and electrochemical characterization data, we elucidate the transformation behavior of these materials under operating conditions. Our findings reveal that NiS2 tends to undergo conversion to nickel hydroxide (Ni(OH)2), while Ni-rich phases like Ni3S4 and NiS exhibit greater stability. Furthermore, we demonstrate that in 3D nanostructures, the initial formation of a protective hydroxide layer on the surface inhibits further sulfide transformation. This nickel sulfide/hydroxide composite shows an improved HER activity with lower onset potential (−0.25 V), Tafel slope (105 mV s−1), and charge transfer resistance (17.3 Ω).

Masked speech recognition by 6–13-year-olds with early-childhood otitis media: effects of acoustic condition and otologic history

Objective To investigate speech recognition in school-age children with early-childhood otitis media (OM) in conditions with noise or speech maskers with or without interaural differences. To also investigate the effects of three otologic history factors. Design Using headphone presentation, speech recognition thresholds (SRTs) were measured with simple sentences. As maskers, stationary speech-shaped noise (SSN) or two-talker running speech (TTS) were used. The stimuli were presented in a monaural and binaural condition (SSN) or a co-located and spatially separated condition (TTS). Based on the available medical records, overall OM duration, OM onset age, and time since the last OM episode were estimated. Study sample 6–13-year-olds with a history of recurrent OM (N = 42) or without any ear diseases (N = 20) with normal tympanograms and audiograms at the time of testing. Results Mixed-model regression analyses that controlled for age showed poorer SRTs for the OM group (Δ-value = 0.84 dB, p = 0.009). These appeared driven by the spatially separated, binaural, and monaural conditions. The OM group showed large inter-individual differences, which were unrelated to the otologic history factors. Conclusions Early-childhood OM can affect speech recognition in different acoustic conditions. The effects of the otologic history warrant further investigation.

Transcriptome analysis reveals the molecular mechanisms underlying the enhancement of salt-tolerance in Melia azedarach under salinity stress

Melia azedarach demonstrates strong salt tolerance and thrives in harsh saline soil conditions, but the underlying mechanisms are poorly understood. In this study, we analyzed gene expression under low, medium, and high salinity conditions to gain a deeper understanding of adaptation mechanisms of M. azedarach under salt stress. The GO (gene ontology) analysis unveiled a prominent trend: as salt stress intensified, a greater number of differentially expressed genes (DEGs) became enriched in categories related to metabolic processes, catalytic activities, and membrane components. Through the analysis of the category GO:0009651 (response to salt stress), we identified four key candidate genes (CBL7, SAPK10, EDL3, and AKT1) that play a pivotal role in salt stress responses. Furthermore, the KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analysis revealed that DEGs were significantly enriched in the plant hormone signaling pathways and starch and sucrose metabolism under both medium and high salt exposure in comparison to low salt conditions. Notably, genes involved in JAZ and MYC2 in the jasmonic acid (JA) metabolic pathway were markedly upregulated in response to high salt stress. This study offers valuable insights into the molecular mechanisms underlying M. azedarach salt tolerance and identifies potential candidate genes for enhancing salt tolerance in M. azedarach.

An invitro nevus explant model for studying the effects of ultraviolet radiation.

Ultraviolet radiation (UVR) has been recognized as a potential trigger for the transformation of benign melanocytic nevi into melanoma. However, the mechanisms governing the formation and progression of melanocytic nevi remain poorly understood. This lack of understanding is partly due to the difficulty in isolating and culturing nevus tissues invitro, resulting in a dearth of robust exvivo models for nevi. Therefore, the establishment of a reliable melanocytic nevus model is imperative. Such a model is essential for elucidating nevus pathogenesis and facilitating the development of effective therapeutic interventions. Therefore, we have sought to establish an exvivo nevus explant model to study UVR stimulation. And the structural integrity and tissue activity of the exvivo nevi explant model was evaluated. We then observed melanogenesis and proliferation activity of the explants after UVR stimulation. There was less blister formation after Day 3 in nevi explants under our modified medium conditions. The nevi explant was able to maintain almost the same morphological structure and tissue activity as invivo tissue within 24 h. Following UVR stimulation, we observed increased melanogenesis and proliferation activity in nevi explants. Nevi explants could serve as an exvivo model for UVR-induced nevi stimulation research.

Optimization studies on laccase activity of Proteus mirabilis isolated from treatment sludge of textile industry factories : Optimization of laccase activity of Proteus mirabilis.

Laccase is an exothermic enzyme with copper in its structure and has an important role in biodegradation by providing oxidation of phenolic compounds and aromatic amines and decomposing lignin. The aim of this study is to reach maximum laccase enzyme activity with minimum cost and energy through optimization studies of Proteusmirabilis isolated from treatment sludge of a textile factory. In order to increase the laccase enzyme activities of the isolates, medium and culture conditions were optimized with the study of carbon (Glucose, Fructose, Sodium Acetate, Carboxymethylcellulose, Xylose) and nitrogen sources (Potassium nitrate, Yeast Extract, Peptone From Soybean, Bacteriological Peptone), incubation time, pH, temperature and Copper(II) sulfate concentration then according to the results obtained. Response Surface Method (RSM) was performed on six different variables with three level. According to the data obtained from the RSM, the maximum laccase enzyme activity is reached at pH 7.77, temperature 30.03oC, 0.5g/L CuSO4, 0.5g/L fructose and 0.082g/L yeast extract conditions. After all, the laccase activity increased 2.7 times. As a result, laccase activity of P. mirabilis can be increased by optimization studies. The information obtained as a result of the literature studies is that the laccase enzymes produced in laboratory and industrial scale are costly and their amounts are low. This study is important in terms of obtaining more laccase activity from P.mirabilis with less cost and energy.

PH Stabilized NH4+-Fed Nutrition Promotes Higher B Uptake and Plant Growth in Rapeseed (Brassica napus L.) by the Upregulation of B Transporters

Rapeseed (Brassica napus L.) is a major oilseed crop with high boron (B) requirements. In hydroponic cultivation, B uptake is highly responsive to the media conditions (e.g., pH). Different N sources, such as NH4+ and NO3−, directly affect the media pH. To date, the underlying mechanisms, the role of the buffering at constant N-sources by separating the effects of pH and/or sources of N-form, for the uptake of B remain indistinct under hydroponic cultivation, particularly in rapeseed. Two identical experiments were conducted using (NH4)2SO4 and Ca(NO3)2 as sole N-sources each set were subjected to four treatments viz. non-buffered, buffered pH 5.0, 6.0, and 7.0 at low (1 µM) and high (100 µM) B for 5 days. The novelty of this study demonstrates that NH4+-fed plants prefer buffered pH (6 and 7) for promoting higher plant growth, which was in line with the B concentration, while NO3−-fed plants were not influenced. These aforesaid findings were further supported by the upregulation of BnaNIP5;1 and BnaBOR1;2 in the root and higher expression of BnaNIP5;1 in shoots with buffered pH in NH4+-fed plants. According to our findings, in NH4+-based buffering, B transport channel protein BnaNIP5;1 (through diffusion) facilitated B uptake and distribution from root to shoot while BnaBOR1;2 mediated active uptake of B was in root under low supply of B. In a nutshell, pH buffering in hydroponics is very important in NH4+-based but not in NO3−-based nutrition for the uptake of boron and growth of the rapeseed plants.

Study on the dynamic response characteristics of lining structures in large-section tunnel blasting using JH-2 model analysis

The lining structures of tunnels are typically constructed using sprayed or cast concrete materials, and their performance and quality during tunnel excavation and blasting are crucial for the stability and safety of tunnels. Therefore, the safe distance between the lining structure and blasting source should be determined to avoid concrete damage caused by blasting vibrations. In this study, taking the subway tunnel of Danshan Station in Qingdao as an example, the JH-2 model is introduced as the constitutive model of the tunnel blasting simulation, and the JH-2 model parameters of the local surrounding rock are obtained by experiments, and finally the numerical simulation and theoretical verification are carried out to study the safety distance of shotcrete under various safety judgment standards. The results indicate that the JH-2 model can effectively simulate the propagation of stress waves under different media conditions, and the closer the strength parameters and pressure constant of the lining structure are to those of the surrounding rock, the safer the concrete–rock bonding interface. During tunnel blasting construction using the ring blasting method, the peak particle velocity (PPV) of the lining structure increases with an increase in the arch angle. Based on the numerical simulation results, we recommend that concrete lining be constructed at a distance of at least 62 m from the blasting source to avoid damage caused by vibrations. The effect of concrete tensile failure caused by longitudinal stress is much smaller than the damage to the bonding interface caused by the PPV and can be neglected.

AI-based engine performance prediction cum advisory system to maximise fuel efficiency and field performance of the tractor for optimum tillage

Tractor engine performance is critical for optimizing energy efficiency in tillage operations across various soil conditions. This study introduces an artificial neural network (ANN) based novel approach for predicting and optimizing specific fuel consumption (SFC) and fuel consumption per tilled area (FCA), during tillage operations. Field tests were conducted to collect the data on rear axle torque, forward velocity, fuel consumption, throttle position, engine speed, and depth of operation under soft, medium, and hard soil conditions while using a reversible MB plough, offset disc harrow, and rigid tyne cultivator. Tractor engine performance was predicted using ANN and optimized by matching the required rear axle power with the available rear axle power. Optimal operating parameters for three distinct tillage implements, each associated with varying soil types and tillage depths, were determined based on minimizing SFC and FCA. An integrated ANN-based user advisory system was developed to provide tractor engine performance predictions and recommendations for optimal operating parameters to maximize fuel efficiency. The recommendations were validated with actual field data, resulting in Mean Absolute Percentage Errors of 7.56% and 8.98% for MB plough, 11.19% and 9.36% for offset disc Harrow, and 10.90% and 10.57% for cultivator, respectively, for SFC and FCA. Highlights ANN-based novel approach is proposed to optimize fuel efficiency during tillage Advisory system is developed to recommend parameters for optimum tillage Suggestions for minimum SFC and FCA are provided in soft, medium and hard soil The accuracy and reliability of advisory system are proven by field validation

Оценка электростатических свойств средств защиты головы, применяемых во взрывопожароопасных зонах

Problems of prevention of explosions and fires caused by the accumulation of electrostatic charge and the following discharge able to ignite an explosion and hazardous medium are well known in global practices. These conditions are specific, inter alia, to engineering industries and determined by the technological necessity to wash or process product surfaces in the medium of flammable liquids. At the same time, explosion and fire safety is primarily ensured by the requirements for non-excess of the minimum value of ignition of medium containing flammable liquids. As a potential source of accumulation of electrostatic charges, personal head protection equipment (safety helmets) required for mechanic safety is considered. Their base and tooling are usually made of polymer materials naturally prone to electrification. The article aims to resolve the problem of assessing the potential accumulation of electric charges during the contact of the scalp with elements of safety helmets. In conditions of current production, the electrostatic potential relative to the earth occurring on the scalp, cheeks, chin, and elements of safety helmet during putting on and off has been studied; the results of the study are provided. The energy of accumulated charges has been determined and compared with the value of minimum energy of ignition of different gasoline brands; conclusions regarding the sufficiency of energy to generate a discharge capable of causing an initiating ignition of flammable liquids have been made. The impact of the length of the employee’s scalp hair on the level of accumulation of electrostatic charges potentially causing ignition of the working medium containing flammable liquids has been determined. The results of the study can be used for the development of a methodology to assess a potential spark hazard occurring due to contact electrification during the use of safety helmets in conditions of explosion and fire-hazardous media and potentially causing fire and explosion.

WITHDRAWN: Accurate and efficient analytical simulation of free vibration for embedded nonlocal CNTRC beams with general boundary conditions

This study aims to evaluate the free vibrational response of restrained nanobeam enriched by nanocomposites based on an exact Fourier series approach. In order to capture the small size effects on the dynamical response, Eringen’s differential form of nonlocal elasticity is used which employs the one size (nonlocal) parameter. Within the framework of Rayleigh and Bernoulli-Euler beam theories to include the effect of nonlocality and by employing Fourier sine series together with Stokes’ transformation, a system of linear equations is obtained then solved by using the coefficient matrix. The combined effects of elastic boundary conditions, elastic medium, dispersion patterns and nonlocal effects are examined by solving this eigen-value problem constructed with Fourier infinite series. Free vibration frequencies are calculated for carbon nanotube reinforced nanobeam under different rigid or restrained boundary conditions, including an elastic medium Winkler-Pasternak type. A comprehensive parametric study is performed, with the particular focus on the influences of distribution pattern, elastic boundary and medium parameter on the free vibrational response of the composite nanobeam with different graded distributions. It is concluded that the addition of a small amount of carbon nanotube material can reinforce the stiffness of the nanobeam, and its free vibration performance is significantly affected by the distribution patterns, elastic medium and boundary conditions.

Establishment of Anoxia/cytokine-induced Injury and Regeneration in Colonoids

Intestinal epithelial cells (IECs) play key roles in nutrient absorption and fluid secretion, providing a barrier to protect cells. IECs, derived from intestinal stem cells (ISCs), are crucial in maintaining intestinal homeostasis and regeneration, which is crucial in gastrointestinal diseases such as inflammatory bowel disease (IBD). To study epithelial injury/regeneration, primary cultured colonoids were utilized as a model system to mimic injury. They retain the composition and differentiation of the tissue, making it a suitable model for studying homeostasis and regeneration of ISCs in vitro. Anoxia and proinflammatory cytokines frequently coexist and exacerbate one another during intestinal injury. However, anoxic conditions have been mainly ignored in previous research. Thus, we developed a regeneration model using cytokines/anoxia injury to better recapitulate intestinal inflammation. Colonoids were treated with cytokines TNF-α and IFN-γ (5 ng/μL) in anoxic conditions for 24 hours to provoke an injury. After 24 hours of injury, the colonoids were exposed to normal oxygen and media conditions for regeneration. Regeneration was observed for 24, 48, and 72 hours and documented using photomicrographs. Morphological analysis detailed that injury was most severe 24 hours post-injury with the combined induction of cytokines and anoxia resulting in decreased colonoid area and number of buds compared to control groups. RT-PCR analysis detailed mRNA expression of stem cell markers ( Lgr5, Bmi1, and Hopx), Wnt ( Axin2, Myc2, and Ascl2), and YAP target genes ( Sca1 and Ctgf) were increased at peak injury and returned to pre-injury levels at 72 hours of recovery. Regeneration of the colonoids is consistent with in vivo observations, thus confirming the validity of the model. This versatile model can be used to study responses to injury and regeneration of intestinal epithelium and the function of ISCs with applications in intestinal disease therapeutics. University of Maryland College of Agriculture and Natural Resources and the Maryland Agricultural Experiment Station Undergraduate Research Grant Grant #4307572 from the Maryland Stem Cell Research Fund. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Growth and organotypic branching of lung-specific microvascular cells on 2D and in 3D lung-derived matrices

Tissue-specific endothelial cells have vital roles in maintenance and functioning of native tissues with constant reciprocal crosstalk with resident cells. Three-dimensional (3D) physio-mimetic in vitro models which incorporate lung-specific microvasculature are needed to model lung-related diseases which involve modulation of endothelial cell behavior like cancer. In this study, we investigated the growth kinetics, morphological changes and responses to biological cues of lung microvasculature on two-dimensional (2D) and in lung matrix-derived 3D hydrogels. HUVEC and HULEC-5a cells were cultured on 2D and compared for their growth, morphologies, and responses to varying growth medium formulations. Brightfield and immunofluorescence imaging was performed to assess differences in morphology. For 3D cultures, native bovine lungs were decellularized, lyophilized, solubilized, and reconstituted into hydrogel form in which endothelial cells were embedded. Cell growth and organotypic branching was monitored in 3D hydrogels in the presence of varying biological cues including lung cancer cell secretome. HUVEC and HULEC-5a cells demonstrated comparable growth and morphology on 2D. However, in 3D lung-derived ECM hydrogels, tissue-specific HULEC-5a cells exhibited much better adaptation to their microenvironment, characterized by enhanced organotypic branching and longer branches. HULEC-5a growth was responsive to lung cancer cell-conditioned medium in both 2D and 3D conditions. In 3D, the concentration of ECM ligand significantly affected cell growth in long-term culture where molecular crowding had an inhibitory role. Our data reveals that HULEC-5a cells offer a reliable alternative to frequently pursued HUVECs with comparable growth and morphology. Due to their intrinsic program for cellular crosstalk with resident cells, the use of tissue-specific endothelium constitutes a vital aspect for modeling physiological and pathological processes. Furthermore, our study is the first demonstration of the synergy between lung-specific microvasculature with lung-specific ECM within a 3D in vitro model.

Exopolysaccharides production by Lactobacillus fermentum under different growth conditions in coconut water medium

Background: Exopolysaccharides (EPS) production gain a lot of attention over recent decades, because EPS can provide beneficial effects, not only on the industrial application but also on the health sector. The understanding of the optimal condition for EPS production will increase the productivity of EPS and can develop EPS with desirable properties. The factors affected EPS production are additional of sugar concentration, temperature, fermentation time, and others. The current work aimed to optimize the utilization of a byproduct leftover of coconut water on the EPS production from Lactobacillus fermentum. Methods: The EPS synthesis were analyzed under various growth conditions in coconut water such as additional of sucrose concentration and incubation times. EPS production of Lactobacillus fermentum was performed by adding 1%, 2%, and 3% of sucrose and 12, 24, 36, and 48 h of incubation periods. The obtained data were analyzed statistically using a two-ways ANOVA test. Results: The EPS production increased as the sucrose concentration and incubation time were increased. The optimal production was found to be in the media supplemented with 3% sucrose and 48 h of incubation period, which gave 12.613 g/L of EPS production. Conclusions: Media of coconut water is suitable for EPS production by Lactobacillus fermentum. Under 3% of additional sucrose concentration and 48 h of incubation time, it produced a larger number of EPS compare to other conditions.

Stainless steels as a solution for corrosion and erosion problems involving grains in agribusiness sector applications

Agricultural activities such as grain processing play a crucial role in Brazil's economy, contributing significantly to its gross domestic product (27.4% in 2021). The use of carbon steel equipment leads to considerable environmental and economic losses in agribusiness due to corrosion. This study explores the erosion-corrosion behavior of a dual-phase 11Cr and an 18Cr8Ni stainless steel (SS), compared to a mild (ASTM A36) and a high-strength carbon (AHSS) steel under conditions close to the agribusiness reality. The impact of corn particles as erosive component associated to pure water and chloride-containing environments was studied using a fog chamber, a topic until now unexplored in the literature. The methodology adopted is novel and explores the erosion-corrosion synergy in cumulative steps, aiming to shed light on the mechanisms of corrosion and wear that affect the materials in the agribusiness industry. The findings reveal that corn particles, despite their soft nature, can easily remove non-adherent corrosion products from the steel surface. In the deionized water medium, the impact of the soft particles can also shift the corrosion pattern of advanced high-strength steels from general corrosion to localized pitting damage reduzing the corrosion rates. The carbon and 11Cr steels showed the highest corrosion rates under the pure corrosion condition in both media. The erosion-corrosion process considerably reduced the corrosion rate, mainly in the deionized water medium for the carbon steels. An exception to this behavior was 18Cr8Ni steel, whose corrosion rates, however, were the lowest among the materials. Low-chromium stainless steel emerges as a cost-effective solution to erosion-corrosion challenges in the agribusiness segment.