Fiber Density Research Articles

Fish oil replacement with different vegetable oils in Onychostoma macrolepis: Effects on fatty acid metabolism based on whole-body fatty acid balance method and genes expression.

To evaluate the fatty acid (FA) metabolism status and possibility as a DHA source of farmed Onychostoma macrolepis, a total of 168 fish (2.03 ± 0.23 g) were fed four diets supplemented with fish oil (FO), linseed oil (LO), soybean oil (SO), and a mixture of LO and SO oil (MO), respectively, for 70 days. Body FA compositions were modified reflecting dietary FAs. Comparing liver and intestine fatty acids with fish fed four diets, the content of ARA in fish fed SO was significantly higher than others (P < 0.05), but showed no difference in muscle. The tissue FA profile showed that the FO-fed group successfully deposited DHA, while the LO-fed group converted ALA to DHA effectively, as well as the liver and intestine EPA was notably highest in the FO group, whereas no difference between the FO and LO group in the muscle. The FA results showed that the DHA contents in the muscle of Onychostoma macrolepis are at a medium-high level compared with several other fish species with the highest aquaculture yield. Correspondingly, in the fish fed diet with LO, SO, and MO, the genes of most FA biosynthesis, transportation, and transcriptional regulation factors were increased in the liver and muscle, but no significant difference was observed in the gene expression of Elovl4b, FATP1, and FABP10 in the muscle. In addition, the enzyme activity involved in PUFA metabolism was higher in fish fed vegetable oil-based diets, corroborating the results of the gene expression. Increased in vivo elongase and desaturase (Δ5, Δ6, and Δ9) activities were recorded in fish fed fish oil-devoid diets, which resulted in the appearance of products associated with elongase and desaturase activities in fish. Besides, as the specific n-3 PUFA synthesis substrate, the dietary supplementation of ALA not only retains most of the nutrition value but also ensures the muscular texture, such as fiber diameter and density. It is concluded that farmed O. macrolepis owns strong n-3 LC-PUFA biosynthetic capacity and high DHA contents so it can be a good DHA source for the population.

Application of multi-layered composite fiber films with enhanced piezoelectric performance in flexible energy harvesting devices

Flexible electronic components such as wearable sensors, energy harvesting devices, and human health monitoring systems based on piezoelectric materials have garnered significant attention within the scientific community. In this study, PZT nanofibers were modified with dopamine hydrochloride prior to their integration into PVDF fibers, serving as piezoelectric reinforcements to enhance the mechanical and piezoelectric characteristics of composite fiber films. Subsequently, a facile hot-pressing method was employed to fabricate multilayered films characterized by a densely packed surface fiber arrangement and a loosely organized internal fiber orientation, which markedly improved the self-supporting ability and fiber density of the electrospun fiber films. A detailed investigation into the micro-morphology, crystallinity, and enhanced piezoelectric properties of the films was conducted. Notably, the composite fiber film containing 6 wt% PZT NFs exhibited a peak piezoelectric coefficient (d33) of 29 pC/N, resulting in an output voltage of 13.8 V. This represents a 2.23-fold enhancement in d33 value and a 4.6-fold increase in output voltage when compared to the pristine PVDF fiber film, which had a d33 of 13 pC/N and an output voltage of 3 V. The power density achieved by the sensor is 0.73 μW/cm2. This performance is underpinned by a high piezoelectric coefficient, outstanding output characteristics, and robust stability, which collectively enhance the sensor's efficacy in various applications. This research presents an effective approach for augmenting the piezoelectric performance of PVDF-based composite films and paves the way for the advancement of piezoelectric energy harvesting applications.

Fibroblast alignment and matrix remodeling induced by a stiffness gradient in a skin-derived extracellular matrix hydrogel

Large skin injuries heal as scars. Stiffness gradually increases from normal skin to scar tissue (20x higher), due to excessive deposition and crosslinking of extracellular matrix (ECM) mostly produced by (myo)fibroblasts. Using a custom mold, skin-derived ECM hydrogels (dECM) were UV crosslinked after diffusion of ruthenium (Ru) to produce a Ru-dECM gradient hydrogel. The Ru diffusion gradient equates to a stiffness gradient and models physiology of the scarred skin. Crosslinking in Ru-dECM hydrogels results in a 23-fold increase in stiffness from a stiffness similar to that of normal skin. Collagen fiber density increases in a stiffness-dependent fashion while stress relaxation also alters, with one additional Maxwell element necessary for characterizing Ru-dECM. Alignment of fibroblasts encapsulated in hydrogels suggests that the stiffness gradient directs fibroblasts to orientate at ∼45 ° in regions below 120 kPa. In areas above 120 kPa, fibroblasts decrease the stiffness prior to adjusting their orientation. Furthermore, fibroblasts remodel their surrounding ECM in a gradient-dependent fashion, with rearrangement of cell-surrounding ECM in high-stiffness areas, and formation of interlaced collagen bundles in low-stiffness areas. Overall, this study shows that fibroblasts remodel their local environment to generate an optimal ECM mechanical and topographical environment. Statement of significanceThis study developed a versatile in vitro model with a gradient stiffness using skin-derived ECM hydrogel with unchanged biochemical environment. Using Ruthenium crosslinking, a 20-fold stiffness increase was achieved as observed in fibrotic skin.The interaction between fibroblasts and matrix depends on changes in the matrix stiffness. The stiffness gradient directed the alignment of fibroblasts with ∼45° in regions with≤ 120 kPa. The cells in regions with the higher stiffness decreased stiffness first and then oriented themselves. Furthermore, fibroblasts remodeled surrounding ECM and regulated its mechanics in a gradient-dependent fashion to reach an optimal condition.Our study highlights the dynamic interplay between cells and surrounding matrix, shedding light on potential mechanisms and strategies to target scar formation and remodeling.

In situ densification and heparin immobilization of bacterial cellulose vascular patch for potential vascular applications

Nowadays, developing vascular grafts (e.g., vascular patches and tubular grafts) is challenging. Bacterial cellulose (BC) with 3D fibrous network has been widely investigated for vascular applications. In this work, different from BC vascular patch cultured with the routine culture medium, dopamine (DA)-containing culture medium is employed to in situ synthesize dense BC fibrous structure with significantly increased fiber diameter and density. Simultaneously, BC fibers are modified by DA during in situ synthesis process. Then DA on BC fibers can self-polymerize into polydopamine (PDA) accompanied with the removal of bacteria in NaOH solution, obtaining PDA-modified dense BC (PDBC) vascular patch. Heparin (Hep) is subsequently covalently immobilized on PDBC fibers to form Hep-immobilized PDBC (Hep@PDBC) vascular patch. The obtained results indicate that Hep@PDBC vascular patch exhibits remarkable tensile and burst strength due to its dense fibrous structure. More importantly, compared with BC and PDBC vascular patches, Hep@PDBC vascular patch not only displays reduced platelet adhesion and improved anticoagulation activity, but also promotes the proliferation, adhesion, spreading, and protein expression of human umbilical vein endothelial cells, contributing to the endothelialization process. The combined strategy of in situ densification and Hep immobilization provides a feasible guidance for the construction of BC-based vascular patches.

Circulating netrin-1 levels are reduced and related to corneal nerve fiber loss in patients with diabetic neuropathy.

Deficiency of neurotropic factors is implicated in diabetic neuropathy (DN). Netrin-1 is a neurotropic factor, but its association with DN has not been explored. We have assessed the association between serum netrin-1 levels and early diabetic neuropathy assessed by quantifying corneal nerve fiber loss using corneal confocal microscopy. A total of 72 participants with type 2 diabetes, without and with corneal nerve fiber loss (DN- n = 42, DN+ n = 30), and 45 healthy controls were studied. Serum netrin-1 levels were measured by enzyme-linked immunosorbent assay, and corneal nerve morphology was assessed using corneal confocal microscopy. Corneal nerve fiber density, branch density, fiber length and serum netrin-1 levels were significantly lower in the DN- and DN+ groups compared with controls (P < 0.001). Netrin-1 levels correlated with corneal nerve fiber length in the DN+ group (r = 0.51; P < 0.01). A receiver operating characteristic curve analysis showed that a netrin-1 cut-off value of 599.6 (pg/mL) had an area under the curve of 0.85, with a sensitivity of 76% and specificity of 74% (P < 0.001; 95% confidence interval 0.76-0.94) for differentiating patients with and without corneal nerve loss. Serum netrin-1 levels show a progressive decline with increasing severity of small nerve fiber damage in patients with diabetes. Netrin-1 could act as a biomarker for small nerve fiber damage in DN.

Association of Uterine Tissue Innervation and Peripheral Nerve Density with Adenomyosis Related Pain. A Systematic Review.

Adenomyosis is associated with dysmenorrhea and chronic pelvic pain; however, the triggering mechanisms of painful stimuli and the role of uterine nerve fibers in the manifestation of pain remain poorly understood. The objective of this study was to systematically review the role of uterine nerve fibers' presence and density in the occurrence of pain in patients with adenomyosis. An electronic search was performed using the Embase, PubMed/Medline, and Cochrane databases. We included all studies from inception to November 2023. A total of ten studies that compared uterine biopsies samples of women with and without adenomyosis were included. The biomarker antiprotein gene product 9.5 was decreased or absent in the endometrium of most included women with adenomyosis. None of the included studies observed a difference in neurofilament (NF) staining between the adenomyosis and non-adenomyosis groups. Studies that assessed nerve growth factor (NGF) staining were heterogeneous in design. One study reported no difference in immunohistochemistry staining in any endometrial layer between the adenomyosis and non-adenomyosis groups, while another reported increased staining in the adenomyosis functional endometrial layer, and a third study reported overexpression of NGF, synaptophysin (SYN), and microtubule-associated protein 2 mRNA in focal adenomyosis alone. Preliminary data from poor-quality studies suggest an increase in the uterine density of nerve fibers in patients with adenomyosis. Well-designed studies are essential to assess the cause-and-effect relationship between uterine nerve fibers and pain in patients with adenomyosis.

The analysis of growth performance and expression of growth-related genes in natural gynogenic blunt snout bream muscle derived from the blunt snout bream (Megalobrama amblycephala, ♀) × Chinese perch (Siniperca chuatsi, ♂)

Gynogenesis is an advanced breeding technique that can quickly establish pure lines, increase progenies genetic variation, cultivate strains with higher dominant traits. As a good model for cross order species hybridization, there are few reports on the study of natural gynogenetic blunt snout bream (GBSB), even on the differences in growth between gynogenesis progenies and their parent fish (blunt snout bream♀ and Chinese perch♂). In this research we compared the growth characteristics of the natural GBSB with its parents. It was found that GBSB had a faster growth rate and higher meat content than its female parent, and the diameter and density of muscle fibers were between those of paternal and maternal fish. We performed transcriptome sequencing of muscle tissues from GBSB and BSB, it identified 1353 significant DEGS in the muscle of the GBSB compared to the BSB, with 820 up- and 533 down-regulated genes. And the relative expression of myosins was notable rise in GBSB. In addition, the MRFs of gene structure, motifs, and phylogenetic relations were characterized in BSB and CP. It showed that MRFs and mstn central domain was highly conserved, but the number of conservative motifs in MRFs of CP is generally higher than that of BSB. Further, the relative expression of myosin complex, myogenic regulatory factors (MRFs), growth hormone/insulin-like growth factor (GH/IGF)-axis and most hypothalamic-pituitary-gonadal (HPG)-axis genes in GBSB were higher than that of the female fish and some were closer to that of the paternal fish. Through the distant hybridization between BSB and CP, a gynogenetic progeny with enhanced growth rate compared to the maternal fish was successfully generated. The research has contributed significant insights into the genes and molecular properties associated with muscle development, as well as their potential functional correlations in response to different functional requirements during the growth process of hybrid progeny formed by cross breeding.

Comparative study on the morphological characteristics, nutritional quality, and tastes of large yellow croaker from five cage culture areas: Relay farming improved fish quality

Relay culture of large yellow croaker from Fujian to Zhejiang is a good model for improved fish quality. However, there is a scarcity of information regarding the comprehensive evaluation of quality alterations following relay culture. In the present study, the large yellow croaker from Ningde (ND) was transferred to net cages in Dongtou (DT), Nanji (NJ), Dongji (DJ) and Dacheng (DC) for continued culture. The 15-month-old fish from ND, DJ, DC, DT, and NJ were sampled on-site for a subsequent comparative study on the morphological characteristics, nutritional quality, and tastes. Results showed that the fish from DJ, DC, DT, and NJ had a lower condition factor (CF) and a higher ratio of body length to height (L/H) than those from ND. The fish from DJ, DC, DT, and NJ, at rates of 77.8%, 91.3%, 95.7%, and 100%, exhibited different geometric morphometrics from those from ND using 10 typical landmarks and Morpho J software. Ralay culture also significantly affected the histology and ultrastructure of muscle fibers, as reflected by an increase in the muscle fiber density and decreases in the ratio of fiber area to gap, I band width, and sarcomere length in the relay culture fish compared to the ND fish. The relay culture fish showed low lipid content and high moisture content. Relay culture also significantly affected the levels of some nutritional composition, including deceases in the levels of hydrolyzed glycine, C16:0, C18:1n9c, C18:2n6c, Σn-6 PUFA (polyunsaturated fatty acids), Fe (iron), Mn (manganese), and Co (cobalt), and increases in the levels of C20:5n3 (EPA, eicosapentaenoic acid), C22:6n3 (DHA, docosahexaenoic acid), C18:0, Σn-3/Σn-6 PUFA, Σn-3 PUFA, Al (aluminum), Se (selenium), Na (sodium), and Ca (calcium). The levels of taste substances such as guanosine monophosphate (GMP) and most flavor amino acids sharply declined after relay culture. However, the ratio of taste activity value (TAV) of ΣDAA to ΣBAA (delicious amino acids/bitter amino acids) in the DT and NJ fish was higher than that in the ND fish. Significant negative relationships between market price and quality parameters were found, including CF, crude lipid, C18:1n9c, C18:2n6c, Σn-6 PUFA, Fe, Mn, and TAV of GMP, whereas there were positive relationships between market price and 4 quality parameters such as L/H, moisture, Σn-3/Σn-6 PUFA, and the ratio of TAC of ΣDAA to ΣBAA. In conclusion, relay culture improved large yellow croaker quality by changes in morphological characteristics, nutritional quality, and tastes, which provides a theoretical basis for quality improvement.

Cutaneous biomarkers of therapeutic efficacy in early treatment of hereditary ATTR amyloid polyneuropathy with tafamidis.

ATTR (ATTRv) amyloidosis neuropathy is characterized by progressive sensorimotor and autonomic nerve degeneration secondary to amyloid deposition caused by a misfolded transthyretin protein (TTR). Small nerve fiber neuropathy is an early clinical manifestation of this disease resulting from the dysfunction of the Aδ and C small nerve fibers. Tafamidis, a selective TTR stabilizer, has proven its efficacy in the earlier stages of hATTR. To evaluate the clinical course and utility of cutaneous pathological biomarkers in patients with ATTR amyloidosis treated with tafamidis compared to control patients. Forty patients diagnosed with early stages of ATTRv amyloidosis (polyneuropathy disability [PND] scores 0-II) underwent small and large nerve fiber neurological evaluations, and annual skin biopsies for intraepidermal nerve fiber density (IENFD) and amyloid deposition index (ADI) estimation. Thirty patients were allocated to receive tafamidis, and 10 patients served as controls. Tafamidis pharmacokinetics analysis was performed in patients who received the treatment. At baseline, 12% of patients in stage PND 0 and 28% in PND I displayed small nerve fiber denervation in the distal thigh, whereas 23% and 38%, respectively, in the distal leg. Similarly, 72% and 84% had amyloid deposition in the distal thigh and 56% and 69% in the distal leg. Following 1 year of treatment, the tafamidis group showed significant clinical improvement compared to the control group, revealed by the following mean differences (1) -9.3 versus -4 points (p = <.00) in the patient's neuropathy total symptom score 6 (NTSS-6) questionnaire, (2) -2.5 versus +2.8 points (p = <.00) in the Utah Early Neuropathy Score (UENS), and (3) +1.2°C versus -0.6 (p = .01) in cold detection thresholds. Among the patients who received tafamidis, 65% had stable or increased IENFD in their distal thigh and 27% in the distal leg. In contrast, all patients in the control group underwent denervation. The ADI either decreased or remained constant in 31% of the biopsies in the distal thigh and in 24% of the biopsies in the distal leg of the tafamidis-treated patients, whereas it rose across all the biopsies in the control group. At the 4-year follow-up, the tafamidis group continued to display less denervation in the distal thigh (mean difference [MD] of -3.0 vs. -9.3 fibers/mm) and the distal leg (mean difference [MD] -4.9 vs. -8.6 fibers/mm). ADI in tafamidis-treated patients was also lower in the distal thigh (10 vs. 30 amyloid/mm2) and the distal leg (23 vs. 40 amyloid/mm2) compared to control patients. Plasma tafamidis concentrations were higher in patients with IENFD improvement and in patients with reduced amyloid deposition. Patients without amyloid deposition in the distal leg at baseline displayed delayed disease progression at 4 years. Cutaneous IENFD and amyloid deposition assessments in the skin of the distal thigh and distal leg are valuable biomarkers for early diagnosis of ATTR amyloidosis and for measuring the progression of small nerve fiber neuropathy. Early treatment with tafamidis slows the clinical progression of the disease, skin denervation, and amyloid deposition in the skin. Higher plasma concentrations of tafamidis are associated with better disease outcomes, suggesting that increasing the drug dose could achieve better plasma concentrations and response rates. This study describes the longest small nerve fiber neuropathy therapeutic trial with tafamidis and is the first to report small fiber symptoms, function, and structural assessments as outcomes.

Influence of a Long Flexible Fiber on the Transport Capability of a Non-Clogging Pump

During the operation of non-clogging pumps, the flexible long fiber is prone to clogging and winding during the flow process, which can result in damage to the non-clogging pump, so a numerical simulation method of a solid–liquid two-phase flow in a non-clogging pump with a flexible long fiber is proposed in this paper. The unsteady numerical simulation of the two-phase flow of a single flexible fiber with different densities, lengths and diameters in a double-blade non-clogging pump was carried out to study the influence of fiber parameters on fiber transport capability. The results show that at a density of 920 kg/m3, 300 kg/m3 and 732 kg/m3, the transport capability of flexible fibers decreases successively, and the transport time T0 is 0.32 s, 0.36 s and 0.48 s, respectively. The transport capability of flexible fibers with a length of 150 mm, 200 mm and 250 mm decreases successively, and the transport time T0 is 0.34 s, 0.48 s and 0.96 s, respectively. The transport time T0 is 0.48 s when the fiber diameter dp is 5 mm. When the fiber diameter dp is 7.5 mm, the transport time T0 is 0.51 s. When the fiber diameter dp is 10 mm, the fiber transport capability of the non-clogging pump decreased significantly, and the transport time T0 is 0.68 s. The fiber length has the most obvious effect on fiber transport capability, followed by the fiber diameter and fiber density.

Loss of Cholinergic and Monoaminergic Afferents in APPswe/PS1ΔE9 Transgenic Mouse Model of Cerebral Amyloidosis Preferentially Occurs Near Amyloid Plaques.

Alzheimer's disease (AD) is characterized by a loss of neurons in the cortex and subcortical regions. Previously, we showed that the progressive degeneration of subcortical monoaminergic (MAergic) neurons seen in human AD is recapitulated in the APPswe/PS1ΔE9 (APP/PS) transgenic mouse model. Because degeneration of cholinergic (Ach) neurons is also a prominent feature of AD, we examined the integrity of the Ach system in the APP/PS model. The overall density of Ach fibers is reduced in APP/PS1 mice at 12 and 18 months of age but not at 4 months of age. Analysis of basal forebrain Ach neurons shows no loss of Ach neurons in the APP/PS model. Thus, since MAergic systems show overt cell loss at 18 months of age, the Ach system is less vulnerable to neurodegeneration in the APP/PS1 model. We also examined whether the proximity to Aβ deposition affected the degeneration of Ach and 5-HT afferents. We found that the areas closer to the edges of compact Aβ deposits exhibit a more severe loss of afferents than the areas that are more distal to Aβ deposits. Collectively, the results indicate that the APP/PS model recapitulates the degeneration of multiple subcortical neurotransmitter systems, including the Ach system. In addition, the results indicate that Aβ deposits cause global as well as local toxicity to subcortical afferents.

Serum neurofilament light chain levels correlate with small fiber related parameters in patients with hereditary transthyretin amyloidosis with polyneuropathy (ATTRv-PN)

BackgroundRecent evidence suggests that both serum neurofilament light chain (sNfL) levels and small fiber related diagnostic variables may be valuable disease biomarkers of hereditary transthyretin amyloidosis with polyneuropathy (ATTRv-PN).Our study aimed to explore the relations between sNfL and small fiber related skin biopsy and quantitative sensory testing (QST) parameters in a cohort of ATTRv-PN patients and pre-symptomatic carriers.MethodsWe retrospectively analyzed data from 13 ATTRv patients and 21 pre-symptomatic carriers who underwent sNfL dosage, skin biopsy, and QST, and analyzed correlations between sNFL, intraepidermal nerve fiber density (IENFD), and cold (CDT) and warm detection thresholds (WDT).ResultsBoth sNfL and small fiber related parameters significantly differed between carriers and patients (sNfL: p < 0.0001; IENFD: p = 0.0008; CDT, WDT: < 0.0001). sNFL levels were normal in all carriers, altered in 85% of patients, negatively correlated with distal IENFD (r = -0.47, p = 0.005), and significantly correlated with CDT (r = -0.68; p < 0.0001) and WDT (r = 0.57; p < 0.0001).ConclusionsOur study showed that sNfL reliably discriminates symptomatic ATTRv-PN patients from pre-symptomatic carriers, and found significant relations between sNfL, skin biopsy, and QST small fiber related parameters, suggesting that sNfL might be a valuable biomarker of peripheral nerve involvement in ATTRv-PN and a supportive criterion for symptomatic disease transition.

Skeletal muscle myosin heavy chain expression and 3D capillary network changes in streptozotocin-induced diabetic female mice.

It is not well-understood how type 1 diabetes (T1DM) affects skeletal muscle histological phenotype, particularly capillarisation. This study aimed to analyze skeletal muscle myosin heavy chain (MyHC) fibre type changes and 3D capillary network characteristics in experimental T1DM mice. Female C57BL/6J-OlaHsd mice were categorized into streptozotocin (STZ)-induced diabetic (n = 12) and age-matched non-diabetic controls (n =12). The muscle fibre phenotype of the soleus, gluteus maximus, and gastrocnemius muscles were characterized based on the expression of MyHC isoforms, while capillaries of the gluteus maximus were assessed with immunofluorescence staining, confocal laser microscopy and 3D image analysis. STZ-induced diabetic mice exhibited elevated glucose levels, reduced body weight, and prolonged thermal latency, verifying the T1DM phenotype. In both T1DM and non-diabetic mice, the gluteus maximus and gastrocnemius muscles predominantly expressed fast-twitch type 2b fibers, with no significant differences noted. However, the soleus muscle in non-diabetic mice had a greater proportion of type 2a fibers and comparable type 1 fiber densities (26.2 ± 14.6% vs 21.9 ± 13.5%) relative to diabetic mice. T1DM mice showed reduced fiber diameters (P = 0.026), and the 3D capillary network analysis indicated a higher capillary length per muscle volume in the gluteus maximus of diabetic mice compared to controls (P < 0.05). Overall, T1DM induced significant changes in the skeletal muscle, including shifts in MyHC fibre types, decreased fibre diameters, and increased relative capillarisation, possibly due to muscle fibre atrophy. Our findings emphasize the superior detail provided by the 3D analytical method for characterizing skeletal muscle capillary architecture, highlighting caution in interpreting 2D data for capillary changes in T1DM.

Bilingual Language Control in the Brain: Evidence from Structural and Effective Functional Brain Connectivity.

Experience in bilingual language control is often accompanied by changes in the structure and function of the brain. Brain structural changes are also often closely related to changes in functions. Previous studies, however, have not directly explored the relationship between structural connectivity and effective functional connectivity of the brain during bilingual language control, and whether the two types of connectivity are associated with behavioral performance of language control. Using behavioral performance, functional, and diffusion imaging techniques, we found that: (1) during language control, the left dorsal lateral prefrontal cortex (dlPFC), left caudate nucleus (CN), inferior parietal lobe, precuneus, and dorsal anterior cingulate cortex (dACC)/pre-SMA were significantly activated. (2) In the language control model with left dlPFC, dACC/pre-SMA, and left CN as ROIs (selected based on activation results and language control models from previous studies), stimuli first enter dACC/pre-SMA and then to left CN. At the left CN, a bidirectional effective connectivity is formed with left dlPFC. (3) There is a nonlinear relationship between effective connectivity during language control and the structural connectivity of the second language learners' brains. Specifically, the fiber density between dACC/pre-SMA and left dlPFC has a positive influence on the bidirectional effective connectivity between left dlPFC and left CN. Findings of the present study contribute evidence toward functional effective connectivity during bilingual language control; toward structural connectivity in the brains of second language learners; as well as toward nonlinear relationships between functional effective connectivity, structural connectivity, and behavioral performance in relation to bilingual language control.

Abstract 123: Hemostatic Fibrin Knob Triggered Microgel To Promote Clotting With Reduced Thrombotic Risks In Neonates

Introduction: Effective neonatal hemostatic therapies are vital for complex surgery-related life-threatening bleeding [1]. Neonatal hemostatic systems differ significantly from adults [2, 3]; however, current treatments for bleeding include adult blood products (ABP). Neonatal ABP use poses an elevated thrombotic risk [4, 5], necessitating alternative approaches. We propose hemostatic therapies leveraging age-dependent clotting differences. In adults, clot formation relies on fibrin(ogen) A:a knob-hole binding, while B:b binding dominates in neonates. Targeting B:b interactions with synthetic Bknob conjugated platelet-like particles (BK-PLP) could crosslink neonatal fibrinogen, catalyze clot formation, and enhance clot strength with lower thrombotic risk than ABP. To test this, we synthesized pNIPAm microgels and conjugated them to B-knob (AHRPYAAK), A-knob (GPRPFPAK), and non-binding control peptides (GPSPFPAK) to prepare BK-PLP, AK-PLP, and NB-PLP. Materials and Methods: We prepared BK-PLP, AK-PLP, and NB-PLP by conjugating microgels with mimetic peptides using EDC-Sulfo-NHS chemistry (Fig. 1A); Fibrin clot polymerization, stability, density, and stiffness, were analyzed via plate-based assays, confocal microscopy, and AFM. Results and Discussion: BK-PLPs significantly increased clot stability compared to AK-PLP, NB-PLP, or fibrin-only clots (Fig. 1B). Fiber density also increased with BK-PLPs, indicating a role in promoting denser fibrin clot structures (Fig. 1C). Clot mechanical properties assessment suggested that AKnob and NB-PLPs may disrupt clot networks, decreasing stiffness (Fig. 1D). Conclusion: Our study explored the effects of peptide conjugate particles on neonatal fibrin clots. BK-PLP incorporation increased clot stability, while AK-PLP and NB-PLP decreased clot stiffness, supporting further BK-PLP development for neonatal hemostasis.

Regional and Sex Differences in the Mouse Colonic Innervation Assessed Using a Computerized Quantitative Approach in Three-dimension of the Enteric Nervous System (ENS)

The ENS consists of the submucosal and myenteric plexus (SMP, MP) and controls a variety of gut functions. The quantitative analysis of innervation in the ENS is still a challenge because the ENS forms a mesh-like three-dimensional (3D) network embedded in the gut wall. We developed a computerized approach to perform direct, objective and automatic measurements of innervation in 3D structures of the mouse colonic ENS. The whole mounts of SMP and MP were prepared along the proximal, transverse and distal colon (pC, tC, dC) collected from 8 naïve mice (C57BL/6J, 6 weeks, 4 of each sex). Each 3D image of SMP or MP was reconstructed from Z-stack confocal images with 50-100 or 30-50 optical sections in the frame 1415×1415 or 708×708 μm and 1 μm apart. The Hu C/D, peripheral form of choline acetyltransferase (pChAT), nitric oxide synthase (nNOS), vasoactive intestinal peptide (VIP), tyrosine hydroxylase (TH) and calcitonin gene-related protein (CGRP) immunoreactive (ir) enteric neurons or fibers were digitally traced and automatically measured with Imaris 9.7 for neuroscientists. The co-localization of pChAT and nNOS was determined with ImarisColoc tool. The densities of neurons and nerve fibers, co-localization of pChAT and nNOS were calculated from 6-8 of 3D images/marker/plexus/segment/mouse (total 2304 3D images) and expressed as the percentage of each type of neurons in total neurons, the volume of each type of nerve fibers in the volume of SMP and MP and co-labeled neurons in each type of neurons. The results demonstrate significant differences in the numbers of two major neuronal populations, their co-localization and densities of three representative nerve fibers among three colonic segments, between two plexuses and sexes. 1. pChAT- and nNOS-ir neurons in the MP: the density of pChAT-ir neurons was 1.4- and 2.1-fold lower in the pC than in the tC and dC (p&lt;0.05, p&lt;0.001), and 1.6- and 4.0-fold higher than that of nNOS-ir neurons in the tC and dC (p&lt;0.001, p&lt;0.001), respectively. In contrast, the density of nNOS-ir neurons was 1.4- or 2.3-fold higher in the pC or tC than dC (p&lt;0.001, p&lt;0.01). 2. Co-localization of pChAT and nNOS in the MP: Co-labeled neurons in the nNOS-ir neurons were 1.4- or 1.5-fold lower in the pC or tC than dC (p&lt;0.05, p&lt;0.05). 3. Nerve fibers: The density of VIP-ir fibers in the SMP was 2.4- and 1.8-fold higher in the pC than tC and dC (p&lt;0.001, p&lt;0.001) while 3.0- and 2.4-fold lower than in the MP in both tC and dC (p&lt;0.001, 0.001). TH-ir fibers in the SMP were 1.6-fold lower in both tC or dC than pC (p&lt;0.01, p&lt;0.01), 1.6-, 2.2- and 1.4-fold lower than MP in the pC, tC and dC (p&lt;0.001, p&lt;0.001, p&lt;0.05). In the MP, they were 1.8- and 1.6-fold lower in the dC than in pC and tC (p&lt;0.001, p&lt;0.001). CGRP-ir fibers did not show significant difference between two plexuses among three segments. 4. Sex differences: The co-labeled neurons in the pChAT-ir neurons were 2.2-, 2.7- and 2.2-fold more in the males than females in three segments (p&lt;0.001, p&lt;001, p&lt;0.001) while those in the nNOS-ir neurons were 1.9-fold more in the males than females only in the dC (p&lt;0.05). VIP-ir fibers in the dC-MP were 1.2-fold higher in the males than females (p&lt;0.05). These data and the novel approach developed in this study will contribute to further assessments of the structure-function relationships and pathological alterations in the colon with mouse models. Supported by NIH/SPARC 1OT2OD024899-01. 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.

Assessment of the association of myofibroblasts and structural components of the extracellular matrix with histopathological parameters of actinic cheilitis and lower lip squamous cell carcinoma.

To evaluate the presence of myofibroblasts (MFs) in the development of lip carcinogenesis, through the correlation of clinical, histomorphometric and immunohistochemical parameters, in actinic cheilitis (ACs) and lower lip squamous cell carcinomas (LLSCCs). Samples of ACs, LLSCCs, and control group (CG) were prepared by tissue microarray (TMA) for immunohistochemical TGF-β, α-SMA, and Ki-67 and histochemical hematoxylin and eosin, picrosirius red, and verhoeff van gieson reactions. Clinical and microscopic data were associated using the Mann-Whitney, Kruskal-Wallis/Dunn, and Spearman correlation tests (SPSS, p < 0.05). ACs showed higher number of α-SMA+ MFs when compared to CG (p = 0.034), and these cells were associated with the vertical expansion of solar elastosis (SE) itself (p = 0.027). Areas of SE had lower deposits of collagen (p < 0.001), immunostaining for TGF-β (p < 0.001), and higher density of elastic fibers (p < 0.05) when compared to areas without SE. A positive correlation was observed between high-risk epithelial dysplasia (ED) and the proximity of SE to the dysplastic epithelium (p = 0.027). LLSCCs showed a higher number of α-SMA+ MFs about CG (p = 0.034), as well as a reduction in the deposition of total collagen (p = 0.009) in relation to ACs and CG. There was also a negative correlation between the amount of α-SMA+ cells and the accumulation of total collagen (p = 0.041). Collagen and elastic density loss was higher in larger tumors (p = 0.045) with nodal invasion (p = 0.047). Our findings show the possible role of MFs, collagen fibers, and elastosis areas in the lip carcinogenesis process.

Physico-chemical, functional, pasting and structural characteristics of two malted maize flours produce under optimal conditions

Maize is a popular gluten-free grain, used in many dishes in Africa and elsewhere. It provides numerous nutrients (especially starch) and contains antinutritional factors (phytates). To reduce these factors and improve the nutritional and physical potential of maize, a number of treatments are required, including malting. This study therefore investigated the physicochemical, functional, structural, and pasting properties of two malted maize varieties (Atp-Y and Coca-sr) produce under optimal conditions. After receiving the two maize varieties from the Agricultural Research Institute for development, the Atp-Y variety was soaked for 30 h at 18.66 °C with 0.37% plant salt, followed by germination at 30 °C for 218.58 h and a ripening period of 40 h, while the Coca-sr variety was soaked for 3.90 h at 20.57 °C with 0.0219% plant salt, followed by germination at 30 °C for 251.55 h and a ripening period of 58.73 h. The flours obtained were subjected to physicochemical, structural, pasting and functional analysis. These analyses showed that the protein, fibre, ash, carbohydrate, starch and energy density content were significantly influenced by the variety. The Atp-Y variety had the highest contents of Ca (855.00 mg/100 g), Na (125.36 mg/100 g) and Cu (0.53 mg/100 g). The availability of Ca, Na and divalent cations was significantly higher in the Atp-Y variety. The Coca-sr variety contributed the most to achieving the protein (40.02 to 73.88%) and lipid (27.30 to 38.08%) requirements of children aged 0 to 59 months. The contents of phytates, cyanides and condensed tannins were similar in both varieties, while oxalates (7.88 mg/100 g), trypsin inhibitors (2.07 mg/100 g) and hydrolyzed tannins (15.02 mg EC/g) were more abundant in the Coca-sr variety. Phenols, flavonoids and saponins ranged between 45.96 and 68.39 mg GAE/g, 32.13 and 39.27 mg CE/g and 0.45 mg/100 g, respectively. No significant differences were found in the physical and functional properties, although the Atp-Y variety had a higher water retention and swelling capacity. Malting variety and conditions had no significant effect on Pasting Time, Setback, Setback ratio and Stability ratio in contrast to Pasting Temperature, Peak viscosity, Holding viscosity, Breakdown and Final viscosity. The Atp-Y grade exhibited the highest pasting parameters when pasting temperature was excluded. Scanning electron microscopy showed that the microscopic structure varied from one variety to another, although the grains had similar sizes. The Microscopic dispersions of grains testifies to the enzymatic activity in the Coca-sr variety, the compact surface layer with protruding grains testifies to the fiber richness of the Atp-Y variety. The grain distribution of the particles showed that more than 50% of the grains in both types were 400 µm in size. X-ray diffraction (XRD) showed that the degradation of the crystalline structure of the starch depended on the variety and the physiological phenomena taking place there. These results clearly show that these flours are well suited for the formulation of foods with high energy and nutritional value. However, they could also be used as sweeteners in panification or as raw materials in the brewing industry.

Abstract 1008: Phenol-Based Denervation Induces Sustained Abdominal Aortic Remodeling

Introduction: Aortic remodeling remains an important contributor to the pathogenesis of aortic disease, including occlusive, aneurysmal, and dissection disease states. The paucity of medical therapies has driven considerable interest in elucidating the pathogenesis of these conditions; new therapeutic targets are critically needed. Prior studies have identified sympathetic nervous system signaling (SNS) as a critical regulator of arterial wall homeostasis with potent effects on inflammation and vascular remodeling. Hypothesis: This study seeks to evaluate the impact of sustained denervation on morphology and wall homeostasis of the abdominal aorta. We hypothesized that topical phenol denervation of the rat abdominal aorta would induce sustained reduction in nerve fiber density with concomitant pathologic changes to tissue morphology. Methods: Male and female Sprague-Dawley rats (n=12), aged 3 months, underwent midline laparotomy for infrarenal aorta exposure. Chemical denervation was induced via a one-time topical application of 10% phenol (n=6), while sham controls received phosphate-buffered saline (n=6). Animals were allowed to recover and subsequently sacrificed after 6 months for analysis encompassing morphology, histology, and immunohistochemistry. Results: At 6 months post-treatment, abdominal aortas subjected to phenol denervation exhibited a significant reduction in nerve fiber density compared to sham controls (92.8 ± 20.3 fibers/mm 2 vs 170.4 ± 79.0 fibers/mm 2 , p=0.042). Denervated aortas demonstrated increased elastin fragmentation breakage scores (2.8 ± 1.1 vs 1.1 ± 0.4, p=0.0056). Phenol denervated rats also had significantly decreased expression of vascular smooth muscle proteins α-SMA (32.6 ± 3.9% vs 24.6 ± 3.3%, p=0.0034) and MYH11(14.2 ± 5.7% vs 21.6 ± 3.5%, p=0.023), as well as elevated adventitial microvascular density (58.9 ± 25.2 vessels/mm 2 vs sham control 18.5 ± 8.7 vessels/mm 2 , p=0.0040). Conclusion: Single-timepoint phenol-based chemical denervation induces sustained alterations in abdominal aortic morphology and vascular remodeling over a 6-month period. These findings underscore the potential of the SNS as a therapeutic target for aortic pathologies.

Omega-3 Fatty Acids Prevent Peripheral Neuropathy and Pro Inflammatory Adipose Tissue Macrophage Phenotype in High-Fat-Fed Mice

Obesity and/or consumption of a high fat diet is associated with the development of peripheral neuropathy (PN) which is characterized by changes in epidermal innervation and increased sensitivity in the extremities. Evidence indicates that omega-3 fatty acids prevent the development of PN and attenuate adipose inflammation, but the interaction of PN and adipose tissue macrophage (ATMΦ) phenotype have not been investigated. We hypothesized that a diet high in unsaturated fatty acids may prevent PN and adipose inflammation, despite the high fat content. PURPOSE: Assess the effects of omega-3 fatty acids on signs of PN and ATMΦ phenotype in high-fat-fed mice. Methods: Male C57Bl/6 mice were randomized to (n = 12-14/group): 10% low fat fish oil (10% FO); 54% high fat fish oil (54% FO); 10% low fat lard (10% L); or 54% high fat lard (54% L) for 14 weeks. PN was characterized by the von Frey test for hindpaw sensitivity at baseline then biweekly through week 14. After 14 weeks, intraepidermal nerve fiber density (IENFD) was quantified by immunohistochemistry of the hindpaw footpad skin using anti-PGP 9.5. The relative percentage of total macrophage cells (F4/80+CD11b+) and M1 (CD206-CD11c+) and M2 (CD206+CD11c-) cell populations in epididymal adipose were identified using flow cytometry. Group differences were identified by 2 x 2 factorial ANOVA. Pearson correlations were used to assess the relationship between hindpaw withdrawal threshold and ATMΦ. Results: 54% L (35.3 ± 1.0 g) had greater body mass compared to all other groups: 10% FO (26.5 ± 0.7, p &lt; 0.001); 10% L (27.8 ± 0.6, p = 0.001); and 54% L (35.3 ± 1.0, p &lt; 0.001). Similarly, fasting insulin was greater in 54% L (3.1 ± 0.4 ng/mL) compared to all other groups: 10% FO (1.0 ± 1.0, p &lt; 0.001); 10% L (1.6 ± 0.2, p &lt; 0.001), and 54% FO (1.0 ± 0.1, p &lt; 0.001). Fasting glucose was greater in 54% L (225.8 ± 7.5 mg/dl) compared to 10% FO (216.1 ± 7.7, p =0.04) and 54% FO (182.6 ± 6.6, p = 0.03), but not 10% L (180.7 ± 17.1, p = 0.483). Following the same pattern as body mass and insulin, homeostatic model assessment of insulin resistance (HOMA-IR) was greater in 54% L (8.1 ± 1.1) compared to all other groups: 10% FO (2.2 ± 0.4, p &lt; 0.001); 10% L (4.1 ± 0.6, p &lt; 0.001); and 54% FO (2.2 ± 0.3, p &lt; 0.001). Hindpaw sensitivity was greater in 54% L (0.08 ± 0.02 g) compared to 54% FO (0.19 ± 0.03, p = 0.02). IENFD was greater in 54% L (52.5 ± 2.2 fibers per mm) compared to 10% FO (44.1 ± 1.5, p = 0.04), but was not different from 10% L (49.7 ± 2.8, p = 0.47) or 54% FO (48.1 ± 3.0, p = 0.24). M1 ATMΦ percentage was greater in 54% L (11.7 ± 2.5 %) compared to all other groups: 10% L (3.8 ± 0.8, p = 0.001), 10% FO (5.2 ± 0.7, p = 0.014), and 54% FO (5.1 ± 0.6 %, p = 0.016). M2 ATMΦ percentage was greater in 10% L (59.7 ± 4.5 %) compared to all other groups: 10% FO (42.1 ± 5.0, p = 0.036), 54% L (41.4 ± 3.9, p = 0.027), and 54% FO (31.4 ± 3.6 %, p &lt; 0.001), but there were no significant differences in M2 ATMΦ percentage in 54% L vs. 54% FO. There were no significant correlations between hindpaw sensitivity and M1 ATMΦ percentage, M2 ATMΦ percentage, or M1:M2 ratio. CONCLUSION: The negative metabolic effects of a high fat lard diet on body mass, glucose, insulin, and insulin resistance were prevented when the dietary fat source was omega-3 fatty acids instead of lard. Notably, mice fed a high fat lard diet had a higher percentage of the pro inflammatory M1 ATMΦ phenotype and developed signs of painful PN including hindpaw hypersensitivity and increased IENFD but these effects were prevented by omega-3 fatty acids in the 54% FO group, despite the high dietary fat content. Supported by the Southern Illinois University Edwardsville Vaughnie Lindsay New Investigator Award. 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.