Number Of Cells Research Articles

Propylparaben-induced endoplasmic reticulum stress triggers G2/M phase cell cycle arrest and initiates caspase-3-dependent apoptosis in human lung cells.

Propylparaben (PrP) is commonly used as an antimicrobial agent in food, cosmetics, and pharmaceuticals. While recent studies have shown that PrP exposure can cause various disruptions in cellular physiology, the precise mechanisms behind these effects remain unclear. In this study, we sought to examine the cytotoxic effects of PrP exposure on human lung cells in a dose- and time-dependent manner. We utilized flow cytometry to analyze the expression of proteins associated with the cell cycle and apoptosis at the single-cell level. Our results showed that PrP treatment leads to a significant upregulation of genes related to ER stress. The activation of ER stress results in a decrease in cyclin B1 levels, which subsequently causes cell cycle arrest at the G2/M phase. After 48h of PrP exposure, the unfolded protein response (UPR) triggers an apoptotic signaling pathway, increasing the number of cells undergoing caspase-3-mediated apoptosis. Together, these physiological changes lead to a reduction in cell viability in the presence of PrP. These findings suggest that PrP exerts harmful effects on human lung cells by activating ER stress, which can lead to apoptosis and cell cycle arrest.

Elucidating the Effects of Formalin as Food Preservative on Hematological Profile and Fertility of Swiss Albino Mice

In Bangladesh, the use of formalin as a food preservative has recently expanded significantly. With this backdrop, the study was attempted to discover changes in the fertility, gross structure, and histoarchitecture of the male reproductive system and hematology in Swiss albino mice as a result of formalin use. Up until experimental week 24, four groups of four mice each were given normal drinking water mixed with formalin at doses of 1%, 5%, 10%, and 20%, respectively, orally. A control group of mice (n=4) were given normal drinking water only. After 24 weeks of treatment, the testicular weight, sperm concentration, morphology, gonadosomatic index (GSI), and hematological profile were evaluated. Testicular histological changes were also evaluated. Formalin-treated mice had significantly lower testes weight, GSI, and sperm count (P<0.05) than the control group. In formalin-treated mice, hematocrit value (HCT) and hemoglobin (Hb) concentration were dramatically reduced; percentage of morphologically healthy epididymal sperm also reduced significantly. In the formalin-treated animals, histopathology indicated degenerative alterations in seminiferous tubules, a lower number of spermatogenic cells, the buildup of spermatozoa with exudates in the tubular lumen, and the displacement of the seminiferous tubule from the basement membrane. According to the findings, formalin has a negative impact on hematological profile and reproductive health of male mice. J. of Sci. and Tech. Res. 6(1): 73-81, 2024

Blood Cell Target Detection Based on Improved YOLOv5 Algorithm

In the medical field, blood analysis is a key method used to evaluate the health status of the human body. The types and number of blood cells serve as important criteria for doctors to diagnose and treat diseases. In view of the problems regarding difficult classification and low efficiency in blood cell detection, this paper proposes an improved YOLOv5-BS blood cell target detection algorithm. The purpose of the improvement is to enhance the real-time performance and accuracy of blood cell type recognition. The algorithm is based on YOLOv5s as the basic network, incorporating the advantages of both CNN and Transformer architectures. First, the BotNet backbone network is incorporated. Then the YOLOv5 head architecture is replaced with the Decoupled Head structure. Finally, a new loss function SIoU is used to improve the accuracy and efficiency of the model. To detect the feasibility of the algorithm, a comparative experiment was conducted. The experiment shows that the improved algorithm has an accuracy of 92.8% on the test set, an average precision of 83.3%, and a recall rate of 99%. Compared with YOLOv8s and PP-YOLO, the average precision is increased by 3.9% and 1%, and the recall rate is increased by 3% and 2%. This algorithm effectively improves the efficiency and accuracy of blood cell detection and effectively improves the problem of blood cell detection.

Sex differentially affects pro-inflammatory cell subsets in adipose tissue depots in a diet induced obesity model

Obesity is a growing pandemic that increases the risk for cardiovascular diseases, type 2 diabetes, and particularly in women also the risk of cancer and neurodegenerative disorders such as dementia and multiple sclerosis. Preclinical studies on obesity focus on male mice as they gain bodyweight faster and show a clear pro-inflammatory phenotype. Here, using male and female mice, we induced obesity by feeding a high fat diet (HFD), and compared adipose tissue (AT) inflammation at the same adiposity stage (% AT/bodyweight) between both sexes. Doing so, we identified that female mice show an increase in the number of pro-inflammatory immune cells in the visceral AT at a lower adiposity stage than male mice, but the effect of HFD is diminished with higher adiposity. Interestingly, only female mice showed an increase in immune cells in the subcutaneous AT after HFD feeding. Nonetheless, we found that pro-inflammatory cytokines in blood plasma mirror the inflammatory stage of the visceral AT in both male and female mice. Uniquely in male mice, myeloid cells in the visceral AT showed a higher inflammasome activation upon HFD. In summary, we showed that adiposity differentially affects immune cells in fat depots based on sex.

Особенности морфофункционального и физиологического состояния осетровых рыб с нефрокальцинозом в условиях садковой аквакультуры

When growing sturgeon fish in cages, which are especially valuable nowadays, great attention should be paid to their health. Along with other pathological manifestations of various natures, a disease such as kidney nephrocalcinosis can pose a potential danger to these aquaculture objects. This pathology was recorded in juvenile sterlet (age – 0+ years), in immature individuals of Russian sturgeon (age – 2 years), in which clinical and pathological examinations were carried out, including pieces of kidneys were selected for histological analysis, blood samples for carrying out hematological and biochemical studies. The appearance of nephrocalcinates in the kidneys of sturgeon fish was confirmed using histological methods: corresponding pathological transformations were found on micropreparations of the kidneys of Russian sturgeon and sterlet. Morphophysiological changes in the condition of the kidneys of the studied sturgeon fish can be assessed as moderate and minor, not life-threatening. Thus, 60% of the examined individuals of Russian sturgeon did not have any detected pathohistological changes in the organ; among sterlet there were 80% of such individuals. When analyzing hematological and biochemical blood parameters, deviations of some indicators were found. Thus, the sterlet had a low level of hemoglobin, all fish had an insufficient number of red blood cells in the blood; in Russian sturgeon, a high level of pathologically altered erythrocytes was determined, the amount of serum protein was increased, deviations were found in the ratio of its fractions, high levels of uric acid, low levels of triglycerides, β-lipoproteins, ALT and AST activity in the blood. Indirectly, these changes in the physiological state of sturgeon fish with nephrocalcinosis indicate that this disease most likely arose as a result of the influence of a complex of factors on the fish, among which the main ones may be disturbances in the hydrochemical regime and the balance of feeding.

LEP O-GlcNAcylation inactivates NF-κB pathway by suppressing LEP protein level and thus mediates cellular senescence and osteogenic differentiation in mouse mesenchymal stem cells

BackgroundCellular senescence is a key driver of decreased bone formation and osteoporosis. Leptin (LEP) has been implicated in cellular senescence and osteogenic differentiation. The aim of this study was to investigate the mechanisms by which LEP mediates cellular senescence and osteogenic differentiation.MethodsC3H10T1/2 cells were treated with etoposide to induce cellular senescence, which was assessed by β-galactosidase staining. Quantitative real-time PCR and western blotting were used to measure the levels of senescence markers p21 and p16, as well as osteogenic differentiation-related genes ALP, COL1A1, and RUNX2. Alkaline phosphatase (ALP) staining and alizarin red S staining were performed to evaluate osteogenic differentiation. The NF-κB pathway and O-GlcNAcylation were assessed by western blotting.ResultsEtoposide treatment increased the number of senescent cells and the levels of p21 and p16, along with elevated LEP expression. These effects were reversed by LEP knockdown. Additionally, LEP knockdown increased ALP staining density and osteoblast mineralization nodules, as well as the mRNA and protein levels of ALP, COL1A1, and RUNX2, indicating that LEP knockdown promoted osteogenic differentiation in C3H10T1/2 cells. Mechanistically, LEP knockdown inactivated the NF-κB pathway by inhibiting the nuclear translocation of p65. Furthermore, OGT was found to promote O-GlcNAcylation of LEP at the S50 site.ConclusionOur findings demonstrated that O-GlcNAcylation of LEP inactivated the NF-κB pathway by reducing LEP protein levels, thereby inhibiting cellular senescence and promoting osteogenic differentiation in C3H10T1/2 cells. This study may provide a novel therapeutic target for the treatment of osteoporosis.

Biocompatibility of light responsive materials prepared for accommodative intraocular lenses manufacturing.

To investigate the biocompatibility and bacterial adhesion properties of light responsive materials (LRM) and analyze the feasibility and biosafety of employing LRM in the preparation of accommodative intraocular lenses (AIOLs). Employing fundamental experimental research techniques, LRM with human lens epithelial cells (hLECs) and human retinal pigment epithelium cells (ARPE-19 cells) were co-cultured. Commercially available intraocular lenses (IOLs) were used as controls to perform cell counting kit-8 (CCK-8), cell staining under varying light intensities, cell adhesion and bacterial adhesion experiments. LRM exhibited a stronger inhibitory effect on the proliferation of ARPE19 cells than commercially available IOLs when co-cultured with the undiluted extract for 96h (P<0.05). Under other culturing conditions, the effects on the proliferation of hLECs and ARPE-19 cells were not significantly different between the two materials. Under the influence of light irradiation at intensities of 200 and 300 mW/cm2, LRM demonstrated a markedly higher inhibitory effect on the survival of hLECs compared to commercially available IOLs (P<0.0001). They also showed a stronger suppressive effect on the survival rate of ARPE-19 cells, with significant differences observed at 200 mW/cm2 (P<0.001) and extremely significant differences at 300 mW/cm2 (P<0.0001). Additionally, compared to commercially available IOLs, LRM had a higher number of cells adhering to their surface (P<0.05), as well as a significantly greater number of adherent bacterium (P<0.0001). LRM, characterized by their excellent non-contact tunable deformability and low cytotoxicity to ocular tissues, show considerable potential for use in the fabrication of AIOLs. These materials demonstrate strong cell adhesion; however, during photothermal conversion processes involving shape deformation under various light intensities, the resultant temperature rise may harm surrounding cells. These factors suggest that while the material plays a positive role in reducing the incidence of posterior capsule opacification (PCO), it also poses potential risks for retinal damage. Additionally, the strong bacterial adhesion of these materials indicates an increased risk of endophthalmitis.

New frontiers in retinal transplantation.

New frontiers about retinal cell transplantation for retinal degenerative diseases start from the idea that acting on stem cells can help regenerate retinal layers and establish new synapses among retinal cells. Deficiency or alterations of synaptic input and neurotrophic factors result in trans-neuronal degeneration of the inner retinal cells. Thus, the disruption of photoreceptors takes place. However, even in advanced forms of retinal degeneration, a good percentage of the ganglion cells and the inner nuclear layer neurons remain intact. This phenomenon provides evidence for obtaining retinal circuitry through the transplantation of photoreceptors into the subretinal region. The eye is regarded as an optimal organ for cell transplantation because of its immunological privilege and the relatively small number of cells collaborating to carry out visual activities. The eyeball's immunological privilege, characterized by the suppression of delayed-type hypersensitivity responses in ocular tissues, is responsible for the low rate of graft rejection in transplant patients. The main discoveries highlight the capacity of embryonic stem cells (ESCs) and induced pluripotent stem cells to regenerate damaged retinal regions. Recent progress has shown significant enhancements in transplant procedures and results. The research also explores the ethical ramifications linked to the utilization of stem cells, emphasizing the ongoing issue surrounding ESCs. The analysis centers on recent breakthroughs, including the fabrication of three-dimensional retinal organoids and the innovation of scaffolding for cell transportation. Moreover, researchers are currently assessing the possibility of CRISPR and other advanced gene editing technologies to enhance the outcomes of retinal transplantation. The widespread use of universally recognized safe surgical and imaging methods enables retinal transplantation and monitoring of transplanted cell growth toward the correct location. Currently, most therapy approaches are in the first phases of development and necessitate further research, including both pre-clinical and clinical trials, to attain favorable visual results for individuals suffering from retinal degenerative illnesses.

Light-Emitting Diode Array with Optical Linear Detector Enables High-Throughput Differential Single-Cell Dielectrophoretic Analysis

This paper presents a lens-free imaging approach utilizing an array of light sources, capable of measuring the dielectric properties of many particles simultaneously. This method employs coplanar electrodes to induce velocity changes in flowing particles through dielectrophoretic forces, allowing the inference of individual particle properties from differential velocity changes. Both positive and negative forces are detectable. The light source utilized in this system is composed of LEDs with a wavelength of 470 nm, while detection is performed using a 256-element optical array detector. Measurements with 10 μm polystyrene beads demonstrate this method can resolve changes equivalent to a Clausius–Mossotti factor of 0.18. Simulations in this work, using values from the literature, predict that Clausius–Mossotti factor differences of 0.18 are sufficient to differentiate viable from nonviable cells and cancerous from multidrug-resistant cancerous cells. We demonstrate that for Chinese hamster ovary (CHO) cells, the method can collect a dielectric response spectrum for a large number of cells in several minutes. We demonstrate that for CHO cells, Clausius–Mossotti factor differences of 0.18 can be discriminated. Due to its simple detection apparatus and the utilization of high-throughput, wide, clog-resistant channels, this method holds promise for a wide range of applications.

Rapid and reproducible generation of glioblastoma spheroids for high-throughput drug screening

Identifying new substances that could potentially be used for tumor therapy and the precise analysis of their spectrum of action requires models that are as similar as possible to the tumor present in the patient. Traditionally, two-dimensional (2D) cell cultures are used. However, these only resemble solid tumors to a limited extent. More realistic in vivo models, such as tissue cultures, which are invaluable for the final analysis of the effect of new substances, are unsuitable for high-throughput screening (HTS), such as substance library screening. Therefore, we addressed which parameters need to be optimized to produce 3D cultures suitable for HTS using established tumor cell lines and ultra-low attachment plates, and we tested which experimental parameters need to be considered. In our studies, we have focused on cell lines from gliomas. Gliomas are incurable tumors of the central nervous system and are the subject of intensive research. Our studies used ten glioma cell lines from which we generated spheroids using ultra-low attachment plates. We then determined the spheroid size as a function of the initial cell number and the culture time. We analyzed cell viability using propidium iodide staining, evaluated the effects of temozolomide and radiation on spheroids, and compared the effect to that on 2D cultures. We found that spheroid size correlated linearly with the initial cell number. Fewer cells (250–500) generally resulted in better growth than a higher number. However, not all cell lines produced growing spheroids at all. The spheroids had an outer layer of living cells and an inner core of dead cells. The size of the inner core of dead cells was different in the various cell lines and developed differently during the incubation period. Radiation affected spheroids more than 2D cultures, especially at higher cell densities. Our results provide insight into using glioma cell lines to form spheroids as model systems. We have identified initial cell numbers as a critical parameter for their effective use in research, offering a hopeful outlook for tumor therapy research and drug development.

Molecular insights into salt tolerance in Dunaliella tertiolectainvolving two betaine aldehyde dehydrogenases.

Understanding salt tolerance mechanisms is crucial for addressing the global challenge of soil salinization and advancing sustainable agricultural practices. Dunaliella tertiolecta, thriving in up to 4.5M NaCl, is a model for studying salt tolerance mechanisms. Two betaine aldehyde dehydrogenase (BADH) genes were identified in D. tertiolecta, namely DtBADH1 and DtBADH2. The phylogenetic analysis revealed that DtBADH1 had similarity to Pseudomonas aeruginosa BADH, while DtBADH2 has high homology to aldehyde dehydrogenase from Chlorella sorokiniana. The 3D models of DtBADH1 and DtBADH2 docking with betaine aldehyde were performed to further validate their binding site, interactions binding the protein and its substrate as well as the conserved amino acids responsible for enzyme activity. We also conducted RNA interference of DtBADH1 and DtBADH2 in D. tertiolecta. Compared to the wild type D. tertiolecta, both BADH-RNAi D. tertiolecta had fewer cell numbers and relatively lower glycine betaine content under high salinity. The findings suggest that both DtBADH1 and DtBADH2 play a crucial role in betaine synthesis, indicating their potential involvement in salt tolerance mechanisms at the molecular level. Additionally, these results highlight D. tertiolecta as a promising candidate for identifying salt stress-responsive genes, which could be utilized for engineering algae or crops to enhance their ability to withstand salinity stress.

Rejuvenating aged osteoprogenitors for bone repair

Aging is marked by a decline in tissue regeneration, posing significant challenges to an increasingly older population. Here, we investigate age-related impairments in calvarial bone healing and introduce a novel two-part rejuvenation strategy to restore youthful repair. We demonstrate that aging negatively impacts the calvarial bone structure and its osteogenic tissues, diminishing osteoprogenitor number and function and severely impairing bone formation. Notably, increasing osteogenic cell numbers locally fails to rescue repair in aged mice, identifying the presence of intrinsic cellular deficits. Our strategy combines Wnt-mediated osteoprogenitor expansion with intermittent fasting, which leads to a striking restoration of youthful levels of bone healing. We find that intermittent fasting improves osteoprogenitor function, benefits that can be recapitulated by modulating NAD+-dependent pathways or the gut microbiota, underscoring the multifaceted nature of this intervention. Mechanistically, we identify mitochondrial dysfunction as a key component in age-related decline in osteoprogenitor function and show that both cyclical nutrient deprivation and Nicotinamide mononucleotide rejuvenate mitochondrial health, enhancing osteogenesis. These findings offer a promising therapeutic avenue for restoring youthful bone repair in aged individuals, with potential implications for rejuvenating other tissues.

Rejuvenating aged osteoprogenitors for bone repair.

Aging is marked by a decline in tissue regeneration, posing significant challenges to an increasingly older population. Here, we investigate age-related impairments in calvarial bone healing and introduce a novel two-part rejuvenation strategy to restore youthful repair. We demonstrate that aging negatively impacts the calvarial bone structure and its osteogenic tissues, diminishing osteoprogenitor number and function and severely impairing bone formation. Notably, increasing osteogenic cell numbers locally fails to rescue repair in aged mice, identifying the presence of intrinsic cellular deficits. Our strategy combines Wnt-mediated osteoprogenitor expansion with intermittent fasting, which leads to a striking restoration of youthful levels of bone healing. We find that intermittent fasting improves osteoprogenitor function, benefits that can be recapitulated by modulating NAD+-dependent pathways or the gut microbiota, underscoring the multifaceted nature of this intervention. Mechanistically, we identify mitochondrial dysfunction as a key component in age-related decline in osteoprogenitor function and show that both cyclical nutrient deprivation and Nicotinamide mononucleotide rejuvenate mitochondrial health, enhancing osteogenesis. These findings offer a promising therapeutic avenue for restoring youthful bone repair in aged individuals, with potential implications for rejuvenating other tissues.

Efficacy analysis of rituximab in treating patients with primary membranous nephropathy dependent on calcineurin inhibitors

BackgroundThis study evaluated the efficacy of rituximab (RTX) in primary membranous nephropathy (PMN) patients with incomplete remission and drug dependence after long-term use of calmodulin inhibitors (CNIs). It aims for complete clinical and immunological remission, and cessation of CNI dependence.MethodsThirty-six patients were enrolled in the study with two groups: drug-dependent and partial remission or immune non-remission group. Both groups underwent RTX therapy with gradual CNI tapering to end CNI dependency and induce complete remission. The primary outcome was overcoming CNI dependency and achieving complete remission after 12 months of RTX therapy. Secondary outcomes included immunological remission and recurrence rates.ResultsThe drug-dependent group (20 patients) achieved significant proteinuria reduction compared to the partial remission or immune non-remission group (16 patients) (P=0.016). After 12 months of RTX treatment, all drug-dependent patients overcame CNI dependency (average withdrawal period: 5.3 ± 3.7 months), with complete remission rates increased from 10% to 70.0% and complete immunological remission rates rose from 35.0% to 90.0%. In the partial remission or immune non-remission group, 14 patients discontinued CNI (average period: 4.6 ± 4.5 months), with complete remission rates increasing from 5.0% to 68.8% and complete immunological remission rates from 6.3% to 68.8%. During follow-up, serum albumin increased, and anti-PLA2R antibodies, 24-hour proteinuria, and CD19+ cell numbers reduced, while creatinine remained stable. Three patients relapsed, four encountered adverse events, and no malignancies or other fatal adverse events were reported.ConclusionsRTX effectively achieves complete clinical and immunological remission in PMN patients dependent on or partially responsive to long-term CNI therapy, reducing recurrence and minimizing prolonged immunosuppressive therapy risks.

Variation in plasmid conjugation among non-typhoidal Salmonella enterica serovars.

Conjugation is a complex phenomenon involving multiple plasmid, bacterial, and environmental factors. Here we describe an IncI1 plasmid encoding multidrug antibiotic resistance to aminoglycosides, sulfonamides, and third generation cephalosporins. This plasmid is widespread geographically and among One Health animal, human, and environmental sectors. We present data on the transmissibility of this plasmid from S. enterica ser. Kentucky into forty strains of S. enterica (10 strains each from serovars Enteritidis, Heidelberg, Infantis, and Typhimurium). Thirty-seven out of forty strains were able to take up the plasmid. Rates of conjugation were variable between strains ranging from 10^-8 to 10^-4. Overall, serovars Enteritidis and Typhimurium demonstrated the highest rates of conjugation, followed by Heidelberg, and then Infantis. No relationships were observed between the recipient cell surface and rate of conjugation. Recipient cell numbers correlated positively with conjugation rate and strains with high conjugation rates had marginally but significantly higher growth parameters compared to strains that took up the plasmid at lower frequencies. Environmental conditions known to impact cell growth such as temperature, nutrient availability, and the presence of antibiotics had a modulating effect on conjugation. Collectively, these results will further understanding of plasmid transmission dynamics in Salmonella which is a critical first step towards the development of mitigation strategies.

N6-methyladenosine modification of THBS1 induced by affluent WTAP promotes Graves’ ophthalmopathy progression through glycolysis to affect Th17/Treg balance

Graves’ ophthalmopathy (GO) obvious manifestation is the imbalance of Th17/Treg. N6-methyladenosine (m6A) methylation is an important regulator of Th17/Treg balance. However, few reports narrate how m6A regulators mediate the role of genes in GO progression. We explored the m6A modification of THBS1 mediated by WTAP, and the mechanism by which THBS1 regulated glycolysis and Th17/Treg balance. A total of 12 peripheral blood (4 GO samples, 4 GH samples, and 4 health samples) were collected to measure the percentage of Th17/Treg in monocytes by flow cytometry. RNA sequencing (RNA-seq) combined with MeRIP sequencing (MeRIP-seq) was used to screen differentially expressed and methylated genes. MeRIP-qPCR was performed to evaluate the m6A abundance of THBS1 after WTAP silencing. Glycolysis of CD4+ T cells was reflected by the lactate content and glucose uptake. The number of Th17 cells was increased in GO peripheral blood, whereas the Treg cells decreased. RNA-seq acquired 679 differentially expressed genes (308 up-regulated, and 371 down-regulated) in the CD4+ T cells of GO compared to healthy control. MeRIP-seq identified 3277 m6A peaks between the GO group and the healthy control group, corresponding with 2744 genes (1143 hypermethylated and 1601 hypomethylated). Combined analysis of RNA-seq and MeRIP-seq showed 81 hypermethylated and up-regulated genes. Among the six candidate genes in the PI3K-signaling pathway, THBS1 was the most significantly differentially expressed and hypermethylated. THBS1 silencing resulted in decreased lactate content and glucose uptake in CD4+ T cells. WTAP was significantly upregulated in CD4+ T cells of GO, and WTAP silencing significantly reduced m6A abundance and expression of THBS1. Upregulated and hypermethylated THBS1 mediated by WTAP promoted glycolysis of CD4+ T cells, affected Th17/Treg balance, and facilitated GO progression. We provided a novel potential target for GO treatment and revealed the molecular mechanism of WTAP and THBS1 in GO under the m6A perspective.

Conditional Tnfaip6-Knockout in Inner Ear Hair Cells Does not Alter Auditory Function.

Noise-induced hearing loss is a worldwide public health issue that is characterized by temporary or permanent changes in hearing sensitivity. This condition is closely linked to inflammatory responses, and interventions targeting the inflammatory gene tumor necrosis factor-alpha (TNFα) are known to mitigate cochlear noise damage. TNFα-induced proteins (TNFAIPs) are a family of translucent acidic proteins, and TNFAIP6 has a notable association with inflammatory responses. To date, there have been few reports on TNFAIP6 levels in the inner ear. To elucidate the precise mechanism, we generated transgenic mouse models with conditional knockout of Tnfaip6 (Tnfaip6 cKO). Evaluation of hair cell morphology and function revealed no significant differences in hair cell numbers or ribbon synapses between Tnfaip6 cKO and wild-type mice. Moreover, there were no notable variations in hair cell numbers or hearing function in noisy environments. Our results indicate that Tnfaip6 does not have a substantial impact on the auditory system.

Effects of Replacing Fishmeal with American Cockroach Residue on the Growth Performance, Metabolism, Intestinal Morphology, and Antioxidant Capacity of Juvenile Cyprinus carpio

Five isonitrogenous and isolipidic diets (Diet 1–Diet 5, with Diet 1 as the control) were formulated to replace 0%, 20%, 40%, 60%, and 80% of fishmeal with American cockroach residue. Juvenile Cyprinus carpio (initial body weight approximately 74 g) were randomly assigned to these diets for a 10-week feeding trial. The Diet 3 group (40% replacement) showed significantly higher final body weight, weight gain rate, specific growth rate, and protein efficiency ratio compared to other groups. No significant differences were observed in crude protein, ash, and total amino acid content across the diets. Groups fed Diet 1 and Diet 2 exhibited higher intestinal trypsin, lipase, α-amylase, and hepatic trypsin activities. Serum triglyceride (TG) levels were highest in the Diet 5 group. Hepatic aspartate aminotransferase (AST) activity was significantly lower in the Diet 3 and Diet 5 groups compared to Diet 1. Serum urea nitrogen levels followed a non-linear trend, initially increasing, then decreasing, and rising again with increasing fishmeal replacement. No significant differences were found in serum total protein (TP) levels among the dietary groups. Intestinal villus number, muscle layer thickness, villus height, villus width, and crypt depth remained consistent across groups. However, goblet cell numbers were significantly reduced at the 60% replacement level, which could impair intestinal barrier function. Diet 3 showed higher serum and hepatic total superoxide dismutase (T-SOD) activity, while Diet 2 had the highest hepatic total antioxidant capacity (T-AOC) activity. Hepatic malondialdehyde (MDA) levels were lowest in the Diet 2 and Diet 5 groups. Immunoglobulin M (IgM) levels showed an increasing trend with higher fishmeal replacement levels. In conclusion, replacing fishmeal with American cockroach residue did not adversely affect growth performance or body composition in juvenile C. carpio. Substituting 20–40% of fishmeal with American cockroach residue enhanced antioxidant capacity and immune function in juvenile C. carpio.

Different dissolved organic matter sources sustain microbial life in a sandy beach subterranean estuary – an incubation study

In subterranean estuaries (STE), fresh and saline groundwater introduce dissolved organic matter (DOM) of different origin. This DOM serves as substrate for microorganisms that thrive in the STE. In high-energy beaches with dynamic porewater advection, microbial communities face frequent changes in groundwater composition, even at several meters depth. It is unknown how DOM from deep STE groundwater (&amp;gt; 5 m depth) is transformed by prevailing microbial communities. To address this question, we performed sediment incubations in flow-through reactors (FTRs) with deep (6 m depth) STE groundwater of low (1.6) and high salinity (29.1). FTR setups were sampled daily for quantification of dissolved organic carbon (DOC), and at start and end (day 13) of the incubation for analysis of DOM composition, microbial cell numbers and community composition. Solid-phase extracted DOM was molecularly characterized via ultrahigh-resolution Fourier-transform ion cyclotron resonance mass spectrometry. Both groundwater types contained mainly reworked DOM. Corroborating its presumed origin, the fresh groundwater had a more terrestrial DOM signature with a higher proportion of aromatic compounds compared to the saline groundwater. Over the course of the incubation, DOC concentrations increased primarily due to leaching of sedimentary organic matter, providing an additional source of DOM. In all setups, the DOM composition changed significantly from start to end, and similarly for fresh and saline groundwater. From the ~2700 molecular formulae (MF) detected on day 0, 34-35% were removed during the incubations, demonstrating the potential of deep STE microbial communities to degrade recalcitrant DOM that is supplied with the advective porewater flow. However, a substantial portion of MF (63-64%) was retained in both groundwater types, indicating that a fraction of deep STE-DOM is resistant to removal. Properties of MF that were newly detected on day 13 (26-28%) were indicative of labile DOM. Some of these newly detected MF were also identified in sediment-leachates, suggesting that beach sediments are a source of fresh DOM for the STE microbial communities. It is likely that due to longer groundwater residence time in the STE, continuous leaching and microbial processing shift the molecular composition of released DOM from more labile to more recalcitrant DOM.

Fibroblasts modulate epithelial cell behavior within the proliferative niche and differentiated cell zone within a human colonic crypt model

Colonic epithelium is situated above a layer of fibroblasts that provide supportive factors for stem cells at the crypt base and promote differentiation of cells in the upper crypt and luminal surface. To study the fibroblast-epithelial cell interactions, an in vitro crypt model was formed on a shaped collagen scaffold with primary epithelial cells growing above a layer of primary colonic fibroblasts. The crypts possessed a basal stem cell niche populated with proliferative cells and a differentiated, nondividing cell zone at the luminal crypt end. The presence of fibroblasts enhanced cell differentiation and accelerated the rate at which a high resistance epithelial cell layer formed relative to cultures without fibroblasts. The fibroblasts modulated cell proliferation within crypts increasing the number of crypts populated with proliferative cells but decreasing the total number of proliferative cells in each crypt. Bulk-RNA sequencing revealed 41 genes that were significantly upregulated and 190 genes that were significantly downregulated in cocultured epithelium relative to epithelium cultured without fibroblasts. This epithelium-fibroblast crypt model suggests bidirectional communication between the two cell types and has the potential to serve as a model to investigate fibroblast-epithelial cell interactions in health and disease.