Disorganized Structure Research Articles

A potential role of a special type of abortive seeds in Cunninghamia lanceolata: promoting the growth of healthy seedlings in active aluminum ions-rich soil

Background and aims“Astringent seed” is a type of abortive seed frequently observed in Chinese fir (Cunninghamia lanceolata). It is widely recognized but poorly understood for its underlying causes. This study investigates the potential of astringent seeds to alleviate the toxic effects of active aluminum ions.MethodsThis study involved treating seeds and seedlings with two distinct concentrations of astringent seeds water extracts under the aluminum ion stress. Then the germination of seeds and growth of seedlings were evaluated and compared.ResultsUnder aluminum stress, both seed germination and seedling growth were notably inhibited. Treatment with a low-concentration of the extract significantly alleviated this inhibition. Root elongation in the seedlings increased by 36.95% compared to the control group, and the aluminum ion accumulation at the root tips was reduced by 38.89% relative to the aluminum-stressed group. This treatment also normalized the levels of malondialdehyde (MDA) in the roots and leaves, enhanced the activities of antioxidative enzymes such as superoxide dismutase (SOD) and catalase (CAT), and restored the levels of endogenous hormones including gibberellin (GA3), indole-3-acetic acid (IAA), methyl jasmonate (Ja-ME), and abscisic acid (ABA). Furthermore, the low-concentration of the extract positively impacted the disorganized chloroplast structures. In contrast, a high-concentration of the extract failed to revert most of these stress indicators.ConclusionLow concentrations of astringent seed water extract effectively alleviate the inhibitory effects of aluminum ions on seed and seedling. This implies that in natural environments, the proximity of healthy seeds to astringent seeds could potentially enhance their growth.

Analysis of Reflectance Confocal Microscopy and Dermoscopic Imaging Features in Diagnosis and Treatment Evaluation of Extramammary Paget’s Disease: A Single-Center Retrospective Study

Objective: Extramammary Paget’s disease (EMPD) is often misdiagnosed as chronic dermatitis or eczema of the scrotum because of its clinical manifestations, leading to delayed diagnosis and treatment. Reflectance confocal microscopy (RCM) and dermoscopy are widely used in clinical practice, enabling early diagnosis and treatment assessment of skin lesions, particularly skin tumors. This study aimed to summarize the RCM and dermoscopic features of EMPD and evaluate treatment outcomes. Methods: We conducted a retrospective analysis of 104 skin lesions from 34 patients diagnosed with EMPD, confirmed through histopathological examination. These patients were treated at our outpatient clinic between January 2022 and December 2023. The analysis focused on typical changes, such as blood vessel patterns and background color under dermoscopy, as well as cellular morphology under RCM. Count data were presented as constituent ratios. Results: Among the 34 patients, 28 were male and 6 were female, and 104 skin lesions were identified. The lesions were classified into three types: 36 red patches, 41 dark red erosive patches, and 27 brown pigmented patches. Dermoscopic examination of EMPD revealed characteristic features, including milky white globules (90.4%, 94/104), irregular shiny white streaks (91.3%, 95/104), dotted blood vessels (91.3%, 95/104), and glomerular-like vessels (88.5%, 92/104). RCM demonstrated epidermal structural disorganization (100%, 104/100), vacuolated cells in the epidermis (94.2%, 98/104), and Paget cells, either scattered or clustered (88.5%, 92/104). The red macular lesions predominantly contained isolated vacuolated cells in the epidermis, the dark red erosive lesions exhibited Paget cell nests in the basal layer, and the brown pigmented lesions contained numerous dendritic cells. These findings are crucial for clinical assessment, ensuring complete tumor removal, and guiding postoperative follow-up. Conclusion: This study identified characteristic dermoscopic and RCM features in patients with EMPD, providing a foundation for non-invasive auxiliary diagnosis. These features facilitate earlier diagnosis and treatment. Additionally, RCM characteristics of different lesion morphologies can help physicians more accurately assess surgical margins, leading to more precise surgical treatment options.

Particle image velocimetry study of the flow control effects of singular and multiple curved serration models

This paper reports on preliminary observations of an investigation of flows associated with models that mimic serrations on the leading edge of barn owl feathers. The objective was to use particle image velocimetry measurements to determine the capacity of singular and multiple curved (three-dimensional) serration models to modify the noise-reducing indicators of a narrow-channeled flow past a cylinder. Four models were tested: 3 singular serration models of respective angles of inclination, α = 24°, 27.5° and 31°, and a model consisting of an array of 3 serrations of α = 24°, 27.5° and 31°. Each case was subjected to flow of Reynolds number (based on the serration height and maximum velocity of the flow) of ∼2,000, simulating the flow regime of local flow around barbs of real barn owl flights. A planar particle image velocimetry technique was used to capture the midspan plane velocities to determine the effects of each model. The results show that using singular serration models of inclination angles than 30° may lead to disorganized spatial structures and enhanced turbulence levels. On the other hand, an array of only 3 curved serrations of different geometries can modify the spatial flow structure into a well-ordered one, resulting in a 50% reduction in turbulence intensities. These initial results suggest that under complex flow conditions, the insertion of single and multiple curved serrations can lead to significant flow changes that may result in potential noise modifications.

The Hexosamine Biosynthetic Pathway alters the cytoskeleton to modulate cell proliferation and migration in metastatic prostate cancer.

Castration-resistant prostate cancer (CRPC) progresses despite androgen deprivation therapy, as cancer cells adapt to grow without testosterone, becoming more aggressive and prone to metastasis. CRPC biology complicates the development of effective therapies, posing challenges for patient care. Recent gene-expression and metabolomics studies highlight the Hexosamine Biosynthetic Pathway (HBP) as a critical player, with key components like GNPNAT1 and UAP1 being downregulated in metastatic CRPC. GNPNAT1 knockdown has been shown to increase cell proliferation and metastasis in CRPC cell lines, though the mechanisms remain unclear. To investigate the cellular basis of these CRPC phenotypes, we generated a CRISPR-Cas9 knockout model of GNPNAT1 in 22Rv1 CRPC cells, analyzing its impact on metabolomic, glycoproteomic, and transcriptomic profiles of cells. We hypothesize that HBP inhibition disrupts the cytoskeleton, altering mitotic progression and promoting uncontrolled growth. GNPNAT1 KO cells showed reduced levels of cytoskeletal filaments, such as actin and microtubules, leading to cell structure disorganization and chromosomal mis-segregation. GNPNAT1 inhibition also activated PI3K/AKT signaling, promoting proliferation, and impaired cell adhesion by mislocalizing EphB6, enhancing migration via the RhoA pathway and promoting epithelial-to-mesenchymal transition. These findings suggest that HBP plays a critical role in regulating CRPC cell behavior, and targeting this pathway could provide a novel therapeutic approach.

Exploring prognostic biomarkers in pathological images of colorectal cancer patients via deep learning.

Hematoxylin and eosin (H&E) whole slide images provide valuable information for predicting prognostic outcomes in colorectal cancer (CRC) patients. However, extracting prognostic indicators from pathological images is challenging due to the subtle complexities of phenotypic information. We trained a weakly supervised deep learning model on data from 640 CRC patients in the prostate, lung, colorectal, and ovarian (PLCO) cancer screening trial dataset and validated it using data from 522 CRC patients in the cancer genome atlas (TCGA) dataset. We created the colorectal cancer risk score (CRCRS) to assess patient prognosis, visualized the pathological phenotype of the risk score using Grad-CAM, and employed multiomics data from the TCGA CRC cohort to investigate the potential biological mechanisms underlying the risk score. The overall survival analysis revealed that the CRCRS served as an independent prognostic indicator for both the PLCO cohort (p < 0.001) and the TCGA cohort (p < 0.001), with its predictive efficacy remaining unaffected by the clinical staging system. Additionally, satisfactory chemotherapeutic benefits were observed in stage II/III CRC patients with high CRCRS but not in those with low CRCRS. A pathomics nomogram constructed by integrating the CRCRS with the tumor-node-metastasis (TNM) staging system enhanced prognostic prediction accuracy compared with using the TNM staging system alone. Noteworthy features of the risk score were identified, such as immature tumor mesenchyme, disorganized gland structures, small clusters of cancer cells associated with unfavorable prognosis, and infiltrating inflammatory cells associated with favorable prognosis. The TCGA multiomics data revealed potential correlations between the CRCRS and the activation of energy production and metabolic pathways, the tumor immune microenvironment, and genetic mutations in APC, SMAD2, EEF1AKMT4, EPG5, and TANC1. In summary, our deep learning algorithm identified the CRCRS as a prognostic indicator in CRC, providing a significant approach for prognostic risk stratification and tailoring precise treatment strategies for individual patients.

Is Achilles tendon structure associated with functional ability and chronic ankle instability in military recruits?

ObjectiveTo determine the relationship between Achilles tendon (AT) structure, functional ability and chronic ankle instability (CAI) in military recruits. MethodsThree hundred and sixty newly recruited infantry male soldiers recruited in April 2022 were assessed for AT structure by Ultrasound Tissue Characterization (UTC), for functional abilities (included proprioception ability, heel-raise test, dynamic postural balance, and hopping agility ability) and for CAI (recurrent sprains and a positive perceived instability). ResultsSoldiers that were identified with disorganized tendon had significantly lower heel-raise and agility scores compared to those with organized tendon structures (33.6 ± 18.1(n) vs. 49.9 ± 28.9(n), p < 0.001; and 5.39 ± 2.12(n) vs. 6.16 ± 1.90(n), p = 0.002, respectively). The best discriminator between soldiers with organized vs. disorganized structure, was heel-raise test (AUC = 0.741). Moreover, soldiers with disorganized AT structure had a higher prevalence of CAI compared with those with organized tendon structures (p < 0.05). ConclusionRecruits with disorganized tendon structures displayed reduced heel-raise score, agility ability and dynamic postural-balance and greater ankle instability. Inferior tendon quality at the onset of military service is an important physical indicator to consider when seeking to manage future injuries and potential for physical performance. Pre-recruitment screening of the AT structure, CAI, and functional abilities, especially in high-intensity infantry programs, needs to be considered.

Urethral tissue characterization using multiparametric ultrasound imaging

A decrease in urethral closure pressure is one of the primary causes of stress urinary incontinence in women. Atrophy of the urethral muscles is a primary factor in the 15 % age-related decline in urethral closure pressure per decade. Incontinence not only affects the well-being of women but is also a leading cause of nursing home admission. The objective of this research was to develop a noninvasive test to assess urethral tissue microenvironmental changes using multiparametric ultrasound (mpUS) imaging technique. Transperineal B-scan ultrasound (US) data were captured using clinical scanners equipped with curvilinear or linear transducers. Imaging was performed on volunteers from our institution medical center (n = 15, 22 to 76 y.o.) during Valsalva maneuvers. After expert delineation of the region of interest in each frame, the central axis of the urethra was automatically defined to determine the angle between the urethra and the US beam for further analysis. By integrating angle-dependent backscatter with radiomic texture feature analysis, a mpUS technique was developed to identify biomarkers that reflect subtle microstructural changes expected within the urethral tissue. The process was repeated when the urethra and US beam were at a fixed angle. Texture selection was conducted for both angle-dependent and angle-independent results to remove redundancies. Ultimately, a distinct biomarker was derived using a random forest regression model to compute the urethra score based on features selected from both processes. Angle-dependent backscatter analysis shows that the calculated slope of US mean image intensity decreased by 0.89 (±0.31) % annually, consistent with the expected atrophic disorganization of urethral tissue structure and the associated reduction in urethral closure pressure with age. Additionally, textural analysis performed at a specific angle (i.e., 40 degrees) revealed changes in gray level nonuniformity, skewness, and correlation by 0.08 (±0.04) %, −2.16 (±1.14) %, and −0.32 (±0.35) % per year, respectively. The urethra score was ultimately determined by combining data selected from both angle-dependent and angle-independent analysis strategies using a random forest regression model with age, yielding an R2 value of 0.96 and a p-value less than 0.001. The proposed mpUS tissue characterization technique not only holds promise for guiding future urethral tissue characterization studies without the need for tissue biopsies or invasive functional testing but also aims to minimize observer-induced variability. By leveraging mpUS imaging strategies that account for angle dependence, it provides more accurate assessments. Notably, the urethra score, calculated from US images that reflect tissue microstructural changes, serves as a potential biomarker providing clinicians with deeper insight into urethral tissue function and may aid in diagnosing and managing related conditions while helping to determine the causes of incontinence.

Experimental model of systemic inflammation during acetaminophen toxicity

Acetaminophen is one of the most toxic drugs that can cause liver damage. At the same time, acetaminophen-induced liver failure is closely associated with the development of systemic inflammatory response syndrome. However, there is no drug aimed at suppressing the systemic inflammatory response. That is, this issue needs to be studied experimentally, but a model of systemic inflammation during acetaminophen overdose has not yet been obtained. Therefore, it was decided to develop an experimental model of systemic inflammation during acetaminophen overdose. The purpose of this study was to experimentally substantiate the semi-lethal dose for acetaminophen overdose in C57Bl/6 mice and to evaluate the readings of blood tests after administration of the drug. To determine the semi-lethal dose, male C57Bl/6 mice were intraperitoneally injected with “Ifimol” (Unique Pharmaceutical Laboratories, India) or acetaminophen solution (Sigma-Aldrich, USA) with a concentration of 14 mg/ml in different doses. When introducing “Ifimol”, it was not possible to achieve a semi-lethal dose. When administering a solution of acetaminophen, 50% mortality was recorded at a dose of 600 mg/kg body weight. After establishing a semi-lethal dose, the experimental group was administered an acetaminophen solution (Sigma-Aldrich, USA) with a concentration of 14 mg/ml at a dose of 600 mg/kg. The control group was injected with saline in an equivalent volume. On the second day, liver and peripheral blood samples were taken. Subsequently, hematological and biochemical blood tests and histological analysis were performed. Histological examination revealed centrilobular necrosis and disorganization of the liver structure. According to the biochemical blood test, the activity of aspartate aminotransferase, alanine aminotransferase, creatinine concentration, and the de Ritis coefficient differed statistically significantly (p 0.05) in the experimental group compared to the control group. Among the hematological blood test parameters, there were statistically significant differences in the number of leukocytes, platelets, as well as the absolute and relative content of granulocytes and lymphocytes. Thus, 48 hours after administration of a semi-lethal dose of acetaminophen, there were signs of damage to internal organs (liver, kidneys) and changes in immune system parameters, which are similar to components of systemic inflammation in humans.

Novel multitarget analgesic candidate SZV-1287 demonstrates potential disease-modifying effects in the monoiodoacetate-induced osteoarthritis mouse model.

Monoiodoacetate (MIA)-induced osteoarthritis (OA) is the most commonly used rodent model for testing anti-OA drug candidates. Herein, we investigated the effects of our patented multitarget drug candidate SZV-1287 (3-(4,5-diphenyl-1,3-oxazol-2-yl) propanal oxime) that is currently under clinical development for neuropathic pain and characterized the mouse model through complex functional, in vivo imaging, and morphological techniques. Knee OA was induced by intraarticular MIA injection (0.5 and 0.8mg). Spontaneous pain was assessed based on weight distribution, referred pain by paw mechanonociception (esthesiometry), edema by caliper, neutrophil myeloperoxidase activity by luminescence, matrix metalloproteinase activity, vascular leakage and bone remodeling by fluorescence imaging, bone morphology by micro-CT, histopathological alterations by semiquantitative scoring, and glia activation by immunohistochemistry. Then, SZV-1287 (20mg/kg/day) or its vehicle was injected intraperitoneally over a 21-day period. MIA induced remarkably decreased thresholds of weight bearing and paw withdrawal, alterations in the tibial and femoral structures (reactive sclerosis, increased trabeculation, and cortical erosions), histopathological damage (disorganized cartilage structure, hypocellularity, decreased matrix staining and tidemark integrity, and increased synovial hyperplasia and osteophyte formation), and changes in the astrocyte and microglia density in the lumbar spinal cord. There were no major differences between the two MIA doses in most outcome measures. SZV-1287 inhibited MIA-induced weight bearing reduction, hyperalgesia, edema, myeloperoxidase activity, histopathological damage, and astrocyte and microglia density. SZV-1287 may have disease-modifying potential through analgesic, anti-inflammatory, and chondroprotective effects. The MIA mouse model is valuable for investigating OA-related mechanisms and testing compounds in mice at an optimal dose of 0.5mg.

PSLBII-30 The effect of different soybean meal batches and protease supplementation on nutrient utilization by broiler chickens

Abstract Soybean meal (SBM) is the major protein source in broiler chicken diets. However, the thermal processing of SBM may have an impact on its protein quality, reducing its solubility when over-processed. In this context, the use of exogenous protease may reduce the negative effects that the over-processing of SBM may have on protein digestibility in broiler chickens. The present study investigated two batches of SBM in broiler chicken diets supplemented or not with protease. One-d-old male Cobb 500 (n = 360) were randomly distributed in a 2 x 2 factorial arrangement (SBM batches: A and B; Protease inclusion: 0 and 50 g/t of diet). SBM protein quality was evaluated through crude protein (CP), KOH solubility, and urease activity test (UA) in batches A and B. Scanning electron microscopy was used to visualize the structural organization of particles from SBM A and B. On d 26, ileal digesta content was collected from 6 broiler chickens that were euthanized to determine the apparent ileal digestibility (AID) of dry matter (DM), crude protein (CP), and ileal digestible energy (IDE). Data obtained were submitted to ANOVA and means were compared by Tukey test with a significance declared of 5% using the GLM procedure on R Software. Protein quality assessment demonstrated KOH solubility of 85% and 75% for SBM from batches A and B, respectively. For UA analysis, SBM batch A presented 0.09 and batch B of 0.01 mg N/g (Table 1). Microscopy images emphasized visual differences in particles from batches A and B. In SBM particles from batch A, disorganized cluster structures and asymmetric pores were observed, whereas in the SBM particles from batch B, insignificant pores and large cluster aggregates were visualized in a denser structure (Figure 1). The SBM batch B presented decreased AID of DM, CP, and IDE (P &amp;lt; 0.05). Protease supplementation improved the AID of CP (P &amp;lt; 0.05) in broilers at 26 d of age, regardless of SBM batches, but no effect was observed for DM and IDE (P &amp;gt; 0.05; Table 2). In conclusion, SBM from batch A had greater values of KOH solubility and UA, suggesting an appropriate thermal process, and ensuring protein quality, which is demonstrated through improvement in nutrient utilization by broiler chickens. The use of 50 g of exogenous protease per t of diet improved the ileal digestibility of CP, regardless of SBM batches.

A novel homozygous splicing mutation in AK7 causes multiple morphological abnormalities of sperm flagella in patients from consanguineous Pakistani families.

Multiple morphological abnormalities of the flagella (MMAF) represent a severe form of sperm defects leading to asthenozoospermia and male infertility. In this study, we identified a novel homozygous splicing mutation (c.871-4 ACA>A) in the adenylate kinase 7 (AK7) gene by whole-exome sequencing in infertile individuals. Spermatozoa from affected individuals exhibited typical MMAF characteristics, including coiled, bent, short, absent, and irregular flagella. Transmission electron microscopy analysis showed disorganized axonemal structure and abnormal mitochondrial sheets in sperm flagella. Immunofluorescence staining confirmed the absence of AK7 protein from the patients' spermatozoa, validating the pathogenic nature of the mutation. This study provides direct evidence linking the AK7 gene to MMAF-associated asthenozoospermia in humans, expanding the mutational spectrum of AK7 and enhancing our understanding of the genetic basis of male infertility.

Exploring the potential of selenium nanoparticles and fabricated selenium nanoparticles @vitamin C nanocomposite in mitigating nicotine-induced testicular toxicity in rats.

The tobacco epidemic signifies a major public health threat. Nicotine (NIC), a major active constituent in tobacco, impedes male fertility and semen quality. This work is implemented to explore the potential of selenium nanoparticles (SeNPs) and the newly fabricated SeNPs @vitamin C (SeNPs@VITC) nanocomposite in mitigating testicular toxicity induced by NIC. The six groups of 48 adult Wistar rats were designed as follows: the control group injected intraperitoneally with normal saline, the SeNPs group treated orally with 2mg/kg of SeNPs, the SeNPs@VITC nanocomposite group treated orally with 2mg/kg of SeNPs@VITC nanocomposite, the NIC group injected intraperitoneally with 1.25mL/kg of NIC, the NIC+ SeNPs group received SeNPs plus NIC, and the NIC+ SeNPs@VITC nanocomposite group received SeNPs@VITC nanocomposite plus NIC. Treatments were administered over a 28-day period. NIC treatment significantly caused poor sperm quality, decreased serum testosterone, increased follicle-stimulating hormone (FSH), luteinizing hormone (LH) concentrations, reduced hemoglobin levels, leukocytosis, disrupted testicular oxidant/antioxidant balance, and disorganized testicular structure. The construction of the novel SeNPs@VITC nanocomposite, compared to NIC plus SeNPs alone, demonstrated a more potent ameliorative effect on NIC-induced reproductive toxicity in adult rats. The SeNPs@VITC nanocomposite significantly increased sperm count, reduced the percentage of sperm head abnormalities, lowered both serum FSH and LH concentrations, and improved the hemoglobin response. Both SeNPs and SeNPs@VITC nanocomposite alleviated the testicular toxicity induced by NIC, but the SeNPs@VITC nanocomposite exhibited superior efficacy. The SeNPs@VITC nanocomposite could be employed to advance enhanced therapeutic strategies for addressing male infertility.

Tendon structure, clinical tests, and pain during-loading in young female competitive gymnasts

ABSTRACT To examine the relationship between Achilles-tendon (AT) and patellar-tendon (PT) structure, clinical-examination and tendon pain in young gymnasts; and, to explore the associations between these factors and age, maturation, and training-load. Two hundred and seventy-four female gymnasts (aged 12.1±1.9 yrs) were assessed for anthropometric measures, pubertal-stage, and training-load. They had clinical-tests (pain-on-palpation for AT and pain-on-palpation and Royal-London Hospital-Test for PT), were asked about tendon-pain during-loading and were assessed for tendon-structure. Gymnasts with positive clinical-tests (with and without pain during-loading) presented a significantly higher prevalence of disorganized AT and PT compared to gymnasts with negative clinical-tests (with and without pain during-loading) (p<0.05). A significant pubertyXpositive clinical-test interaction was found for disorganized PT structure, whereby a disorganized structure was more prevalent among post-pubertal gymnasts with positive clinical-tests compared to pre-pubertal participants with negative clinical-tests (F(1, 263)=9.436, p=0.002). In gymnasts with positive clinical-tests, significant correlations were found between disorganized AT and PT structures and age, and training-load (p<0.05). An increased prevalence of disorganized tendon structure (regardless of pain during-loading) was seen in participants with positive clinical-tests. This disorganized tendon-structure was found to be significantly related to increased age, post-pubertal stage, and higher training hours in gymnasts with positive clinical-tests.

Titanium nanoparticles released from orthopedic implants induce muscle fibrosis via activation of SNAI2

Titanium alloys represent the prevailing material employed in orthopedic implants, which are present in millions of patients worldwide. The prolonged presence of these implants in the human body has raised concerns about possible health effects. This study presents a comprehensive analysis of titanium implants and surrounding tissue samples obtained from patients who underwent revision surgery for therapeutic reasons. The surface of the implants exhibited nano-scale corrosion defects, and nanoparticles were deposited in adjacent samples. In addition, muscle in close proximity to the implant showed clear evidence of fibrotic proliferation, with titanium content in the muscle tissue increasing the closer it was to the implant. Transcriptomics analysis revealed SNAI2 upregulation and activation of PI3K/AKT signaling. In vivo rodent and zebrafish models validated that titanium implant or nanoparticles exposure provoked collagen deposition and disorganized muscle structure. Snai2 knockdown significantly reduced implant-associated fibrosis in both rodent and zebrafish models. Cellular experiments demonstrated that titanium dioxide nanoparticles (TiO2 NPs) induced fibrotic gene expression at sub-cytotoxic doses, whereas Snai2 knockdown significantly reduced TiO2 NPs-induced fibrotic gene expression. The in vivo and in vitro experiments collectively demonstrated that Snai2 plays a pivotal role in mediating titanium-induced fibrosis. Overall, these findings indicate a significant release of titanium nanoparticles from the implants into the surrounding tissues, resulting in muscular fibrosis, partially through Snai2-dependent signaling.Graphical

Comparative effect of ivermectin and amitraz on cellular architecture of ovaries, synganglion and Gené's organ of Rhipicephalus microplus (Acari: Ixodidae).

The present study investigated the effect of ivermectin and amitraz on the cellular architecture of vital organs of Rhipicephalus microplus. Adult female ticks were treated with lethal concentrations (LC95) of ivermectin and amitraz, and the ovaries, synganglion, and Gené's organ were processed 48h post treatment. In both the treatment groups, the ultra-thin sections of ovary exhibited deformed oocytes, irregular plasmic membrane and chorion layer, extensive vacuolation in the cytoplasm mainly at periphery of the cell and oocyte-pedicel junction. Marked vacuolations in the cortex and neuropile region with significant structural disorganization of the neural fibers were common alterations observed in the synganglion of ticks exposed to ivermectin and amitraz. The tissue sections of Gené's organ revealed deformed tubular glands with severe loss of cellular limit of secretory epithelium and cytoplasmic vacuolations in the ivermectin treated ticks whereas, the alterations were comparatively less severe in amitraz exposed ticks. The cellular deformities in these vital organs probably impaired reproductive function, nerve signal transmission and metabolic activities and thus affected fecundity and survivability of the treated ticks. The findings suggested that the action of ivermectin and amitraz are not restricted to the nervous system of ticks, but also on other vital organs, ovary and Gené's organ affecting the oviposition. The study provided insights into the development of targeted interventions for tick control strategies.

Silybin attenuates avermectin-induced oxidative damage in carp respiration by modulating the cGAS-STING pathway and endoplasmic reticulum stress.

Avermectin is a commonly used insect repellent for aquaculture and crops, but it is easy to remain in the aquatic environment, causing organism disorders, inflammation, and even death. This resulted in significant economic losses to the carp aquaculture industry. Silybin has antioxidant, anti-inflammatory, and anti-apoptotic properties. However, it is unclear whether Silybin counteracts gill damage caused by avermectin exposure. Therefore, we modeled avermectin exposure and Silybin intervention by adding 2.404 μg/L avermectin to water and 400 mg/kg of Silybin to feed. Gill tissue was collected and analyzed in depth during a 30-day experimental period. The results showed that avermectin exposure induced structural disorganization of gill filaments and led to increased reactive oxygen species, inhibition of antioxidant functions, induction of inflammatory responses, and endoplasmic reticulum stress in addition to the endogenous apoptotic pathway. In contrast, Silybin effectively alleviated pathological changes and reduced reactive oxygen species levels, thereby attenuating oxidative stress and endogenous apoptosis and inhibiting endoplasmic reticulum stress pathways. In addition, Silybin reduced avermectin-induced gill tissue inflammation in carp, and it is considered that it might modulate the cGAS-STING pathway. In summary, Silybin alleviates avermectin-induced oxidative damage within the carp's respiratory system by modulating the cGAS-STING pathway and endoplasmic reticulum stress. The main goal is to understand how Silybin reduces oxidative damage caused by avermectin in carp gills, offering management strategies. Concurrently, the current study proposes that Silybin can serve as a dietary supplement to reduce the risks brought on by repellent buildup in freshwater aquaculture.

A novel homozygous missense TTC12 variant identified in an infertile Pakistani man with severe oligoasthenoteratozoospermia and primary ciliary dyskinesia.

TTC12 is a cytoplasmic and centromere-localized protein that plays a role in the proper assembly of dynein arm complexes in motile cilia in both respiratory cells and sperm flagella. This finding underscores its significance in cellular motility and function. However, the wide role of TTC12 in human spermatogenesis-associated primary ciliary dyskinesia (PCD) still needs to be elucidated. Whole-exome sequencing (WES) and Sanger sequencing were performed to identify potentially pathogenic variants causing PCD and multiple morphological abnormalities of sperm flagella (MMAF) in an infertile Pakistani man. Diagnostic imaging techniques were used for PCD screening in the patient. Real-time polymerase chain reaction (RT‒PCR) was performed to detect the effect of mutations on the mRNA abundance of the affected genes. Papanicolaou staining and scanning electron microscopy (SEM) were carried out to examine sperm morphology. Transmission electron microscopy (TEM) was performed to examine the ultrastructure of the sperm flagella, and the results were confirmed by immunofluorescence staining. Using WES and Sanger sequencing, a novel homozygous missense variant (c.C1069T; p.Arg357Trp) in TTC12 was identified in a patient from a consanguineous family. A computed tomography scan of the paranasal sinuses confirmed the symptoms of the PCD. RT-PCR showed a decrease in TTC12 mRNA in the patient's sperm sample. Papanicolaou staining, SEM, and TEM analysis revealed a significant change in shape and a disorganized axonemal structure in the sperm flagella of the patient. Immunostaining assays revealed that TTC12 is distributed throughout the flagella and is predominantly concentrated in the midpiece in normal spermatozoa. In contrast, spermatozoa from patient deficient in TTC12 showed minimal staining intensity for TTC12 or DNAH17 (outer dynein arms components). This could lead to MMAF and result in male infertility. This novel TTC12 variant not only illuminates the underlying genetic causes of male infertility but also paves the way for potential treatments targeting these genetic factors. This study represents a significant advancement in understanding the genetic basis of PCD-related infertility.

#2695 A Polycystin-1-Bicaudal C1 complex regulates focal adhesion function through integrin beta-1, F-actin organisation and local translation in HKE cells

Abstract Background and Aims Variants in Bicc1, result in a recessive murine PKD phenotype (bpk, jcpk) by uncertain mechanisms. In a recent study, we reported genetic and biochemical evidence of novel interactions between PKD1, PKD2 and BICC1 in Xenopus, mouse and man suggesting that BICC1 is a modifier of the ADPKD phenotype (submitted). In this study, our major aim was to investigate the molecular mechanisms underlying cystogenesis that result from loss of the normal interaction between the PKD1 protein, Polycystin-1 (PC1), a cell surface receptor and Bicaudal C1 (BICC1), an RNA-binding protein. Methods Isogenic PKD1 and BICC1 CRISP-Cas9 KO cells were generated from a conditionally-immortalised wild-type human proximal tubular line (UCL93); RNA-seq was performed in PKD1 and BICC1 KO cells to identify commonly altered genes and pathways; MS pull-down was conducted in HEK293 cells with BICC1-HA to identify BICC1 interacting proteins; cellular phenotypes in normal and KO cells were assessed through adhesion and migration assays, alongside Traction Force Microscopy (TFM); proximal ligation assays (PLA) to confirm protein-protein interactions; single-molecule FISH (smFISH) to localize mRNA transcripts; puromycin labeling with a proximity ligation assay (Puro-PLA) to detect sites of translation of specific mRNAs. Results Consistent with a functional interaction, loss of PC1 resulted in lower BICC1 levels and vice-versa in single KO (SKO) cells. In migrating cells, BICC1 was expressed at the leading edge where it co-localised with β1-integrin (ITGB1) by PLA, suggesting enrichment at focal adhesion sites (FA). BICC1 KO cells had no cilia phenotype but instead displayed striking disorganization of stress fibres and aster-like structures, features which could be completely rescued by lentiviral Bicc1-HA re-expression. Gene set enrichment and pathway analysis confirmed that F-actin organization and actin-associated proteins (AAP) were commonly dysregulated in SKO cells. BICC1 was also found to interact ITGB1, several components of an integrin-associated complex and proteins involved in actin turnover by MS pull-down. Both SKO cells showed major abnormalities of cell attachment, detachment and directional migration indicative of functional alterations in cell-matrix and cell-cell adhesion. We confirmed that FA structure and function were altered by reduced co-localisation of several AAPs (including TNS1 and FBLIM1) and by TFM. BICC1 directly interacted with F-actin and FBLIM1 in pull-down assays and with FBLIM1 by RNA-IP. Finally, we found evidence of FBLIM1 mRNA by smFISH and of local FBLIM1 translation (Puro-PLA) at FA sites in wild-type cells, both of which were markedly reduced in SKO cells. Conclusion Our data reveal the key roles for actin dynamics and local translation as novel mechanisms regulated by PC1 and BICC1 in the pathogenesis of ADPKD. We demonstrate that BICC1 and PC1 are co-regulated, forming a PC1-BICC1 complex critical for maintaining the structural integrity and function of FAs in normal kidney epithelial cells. Loss of this interaction results in major alterations in cell adhesion, migration and stress fibre organisation, features typical of the ADPKD cellular phenotype. BICC1 plays a dual role in this process via protein-protein interactions with ITGB1, F-actin and other AAPs, and through protein-RNA interactions regulating the local translation of key AAPs (FBLIM1) at FA sites.

Protective effect of zinc gluconate on intestinal mucosal barrier injury in antibiotics and LPS-induced mice

ObjectiveThe aim of the study is to investigate the function and mechanism of Zinc Gluconate (ZG) on intestinal mucosal barrier damage in antibiotics and Lipopolysaccharide (LPS)-induced mice.MethodsWe established a composite mouse model by inducing intestinal mucosal barrier damage using antibiotics and LPS. The animals were divided into five groups: Control (normal and model) and experimental (low, medium, and high-dose ZG treatments). We evaluated the intestinal mucosal barrier using various methods, including monitoring body weight and fecal changes, assessing pathological damage and ultrastructure of the mouse ileum, analyzing expression levels of tight junction (TJ)-related proteins and genes, confirming the TLR4/NF-κB signaling pathway, and examining the structure of the intestinal flora.ResultsIn mice, the dual induction of antibiotics and LPS led to weight loss, fecal abnormalities, disruption of ileocecal mucosal structure, increased intestinal barrier permeability, and disorganization of the microbiota structure. ZG restored body weight, alleviated diarrheal symptoms and pathological damage, and maintained the structural integrity of intestinal epithelial cells (IECs). Additionally, ZG reduced intestinal mucosal permeability by upregulating TJ-associated proteins (ZO-1, Occludin, Claudin-1, and JAM-A) and downregulating MLCK, thereby repairing intestinal mucosal barrier damage induced by dual induction of antibiotics and LPS. Moreover, ZG suppressed the TLR4/NF-κB signaling pathway, demonstrating anti-inflammatory properties and preserving barrier integrity. Furthermore, ZG restored gut microbiota diversity and richness, evidenced by increased Shannon and Observed features indices, and decreased Simpson’s index. ZG also modulated the relative abundance of beneficial human gut bacteria (Bacteroidetes, Firmicutes, Verrucomicrobia, Parabacteroides, Lactobacillus, and Akkermansia) and harmful bacteria (Proteobacteria and Enterobacter), repairing the damage induced by dual administration of antibiotics and LPS.ConclusionZG attenuates the dual induction of antibiotics and LPS-induced intestinal barrier damage and also protects the intestinal barrier function in mice.

Conjugates of glycosylated antarctic krill proteins and curcumin: Maintaining the storage quality of salmon fillets coupling with photodynamic inactivation

Antarctic krill proteins (AKP) were successfully glycosylated by inulin driven by ultrasound (300 W, 20 min) and pH shift (11.0) to prepare the conjugates of glycosylated AKP and curcumin. The preservation effects of the conjugates-mediated photodynamic inactivation (PDI) on the storage quality of salmon fillets were investigated. Results showed that the glycosylated AKP (AKP-pH11-U20) exhibited a significantly increased solubility, and a relatively small and loose morphology characterized by SEM, an unfolded and disorganized structure by FTIR and intrinsic fluorescence analysis, and a decreased surface hydrophobicity by bromophenol blue staining, and all these changes contributed to its reduced surface tension. On this basis, the glycosylated AKP efficiently loaded curcumin (323.4 μg/mg) to form the conjugates, and the conjugates-mediated PDI greatly inhibited 99.8% bacterial cells and the migration of water, retarded the change of color and pH, suppressed the lipid oxidation and degradation during 4 °C storage for 7 days, compared to the conventional curcumin-mediated PDI. This study expands the application of new marine protein resources in preserving the quality of aquatic products.