LF Group Research Articles

Schizochytrium limacinum altered antioxidant capacity and transcriptome profiles in Pacific white shrimp fed a low-fishmeal diet

Dietary low-fishmeal (FM) have negative impacts on growth performance and metabolic responses in aquatic animals, while Schizochytrium limacinum have multiple benefits due to its rich contents of docosahexaenoic acid (DHA). However, little information is available on the effects of S. limacinum on Pacific white shrimp (Litopenaeus vannamei) in a low-FM diet (LF). An 8-week feeding trial were used to investigate the impacts of S. limacinum on growth performance, as well as morphological changes, biochemical indicators, antioxidant capacity, and transcriptomic profiles of hepatopancreas in shrimp fed a low-FM diet. Compared to LF diet, 0.3% S. limacinum (LFLD) diet significantly decreased the specific growth rate (SGR), and did not affect the feed conversion rate (FCR) and weight gain rate (WGR); whereas 0.6% S. limacinum (LFHD) diet did not affect the SGR, FCR and WGR. Histological structures of hepatopancreas showed that LFLD and LFHD treatments reduced the number and size of secretory cells and increased the number of storage cells, compared to LF group. LFLD and LFHD diets significantly decreased the concentrations of triglyceride and total cholesterol, significantly increased the levels of glutathione peroxidase and total antioxidative capacity, while did not affect the levels of malondialdehyde and superoxide dismutase. A total of 135 differential expression genes (DEGs) were identified in LFLD group and 108 DEGs were identified in LFHD group (both compared to LF group). Administration of S. limacinum altered the transcriptomic profiles, while LFHD induced a pattern similar to that of LF. LFLD and LFHD treatments mainly affected the 9 KEGG metabolic pathways, including “Arachidonic acid metabolism” and “Glutathione metabolism”. In conclusion, LFLD treatment impaired the growth performance of shrimp, while LFHD treatment did not affect it; and both treatments improved the morphological structure, lipid accumulation and antioxidant capacity of hepatopancreas.

Effect of delayed distal radius fracture fixation on the difficulty of surgical operation

PurposeThe effect of delayed distal radius fracture (DRF) fixation by volar locking plates (VLPs) on the operative time, rate of bone graft use and need for additional surgical incisions was analyzed. Short- and mid-term outcomes were compared between early and delayed DRF fixation. MethodsA retrospective cohort review of DRFs treated with VLPs was performed. The effects of delayed fixation were analyzed by 1) comparing intraoperative and radiographic parameters and follow-up outcomes between early fixation (EF, 1–10 days) and late fixation (LF, ≥11 days), 2) predicting the prolonged operative time using linear regression analysis, and 3) predicting the rate of bone graft use and additional incision using odds ratios. ResultsOf 104 patients, 51 and 53 were in the EF and LF groups, respectively. EF showed a significantly shorter operative time, lower rate of bone grafting and fewer additional incisions (94.80 vs. 123 minutes, 3.92% vs. 26.18% and 2% vs. 20.45%, respectively). Radiographic parameters, immediate postoperative pain and wrist range of motion were significantly better in the EF group. However, the visual analog scale (VAS) pain and Patient-Rated Wrist Evaluation (PRWE) scores were not different at the 12-month follow-up. For every day of delay, the operation was prolonged by 2.17 min, the rate of bone grafting increased by 8%, and the chance of additional incisions increased by 8%. ConclusionDelayed DRF fixation affects intraoperative and postoperative outcomes. We encourage internal fixation for DRFs as early as possible to reduce surgical complexity and improve postoperative outcomes.

Evaluation of lipidomic change in goat sperm after cryopreservation.

The current study aimed to detect the relationship between the spermatozoa cryotolerance and the post-thawed sperm lipidome. Ejaculates from 20 goats, and performed a uniform frozen-thawed procedure in this study. According to the total motility of thawed sperm of goats, semen samples were classified into HF group (High Freezers, n = 8) with >60% total motility and LF group (Low Freezers, n = 8) with < 45% total motility. The lipidomic analysis based on UHPLC-MS/MS was utilized to investigate the relationship between sperm cryotolerance and their lipid metabolites expression. The results showed that the cryotolerance of sperm from different individual goats were in great variation. The total motility of post-thawed sperm in HF group (60.93 ± 2.43%) is significantly higher than that in LF group (34.04 ± 3.41%, P < 0.01). And the post-thawed sperm in HF group exhibited significantly higher plasma membrane (59.06 ± 2.34%) and acrosome integrity (62.93 ± 1.15%) than that in LF group (34.06 ± 4.85%, 44.92 ± 2.19% respectively, P < 0.01). The total of 29 lipid subclasses and 1,133 lipid molecules in the post-thawed goat sperm were identified by lipidomics analysis. The lipid content of thawed sperm in HF group was higher than that in LF group, the lipid profile in HF group was significantly separated from LF group, which indicated that the difference in lipid composition and lipid metabolism mode of sperm between the two groups was existed, especially the expression of phosphatidylcholine and triglyceride molecules. In conclusion, the cryotolerance of sperm from different individual goats were in great variation. Sperm with high cryotolerance may be able to uptake more lipids during cryopreservation. The increase in phosphatidylcholine and triglyceride content of thawed. Sperm may relate to more active lipid anabolic processes.

Identification of differentially expressed mRNAs and miRNAs in spermatozoa of bulls of varying fertility.

Bulls used in artificial insemination, with apparently normal semen quality, can vary significantly in their field fertility. This study aimed to characterize the transcriptome of spermatozoa from high (HF) and low (LF) fertility bulls at the mRNA and miRNA level in order to identify potential novel markers of fertility. Holstein-Friesian bulls were assigned to either the HF or LF group (n = 10 per group) based on an adjusted national fertility index from a minimum of 500 inseminations. Total RNA was extracted from a pool of frozen-thawed spermatozoa from three different ejaculates per bull, following which mRNA-seq and miRNA-seq were performed. Six mRNAs and 13 miRNAs were found differentially expressed (P < 0.05, FC > 1.5) between HF and LF bulls. Of particular interest, the gene pathways targeted by the 13 differentially expressed miRNAs were related to embryonic development and gene expression regulation. Previous studies reported that disruptions to protamine 1 mRNA (PRM1) had deleterious consequences for sperm chromatin structure and fertilizing ability. Notably, PRM1 exhibited a higher expression in spermatozoa from LF than HF bulls. In contrast, Western Blot analysis revealed a decrease in PRM1 protein abundance for spermatozoa from LF bulls; this was not associated with increased protamine deficiency (measured by the degree of chromatin compaction) or DNA fragmentation, as assessed by flow cytometry analyses. However, protamine deficiency was positively and moderately correlated with the percentage of spermatozoa with DNA fragmentation, irrespective of fertility group. This study has identified potential biomarkers that could be used for improving semen quality assessments of bull fertility.

Muscle contractile exercise through a belt electrode device prevents myofiber atrophy, muscle contracture, and muscular pain in immobilized rat gastrocnemius muscle.

Immobilization of skeletal muscles causes muscle atrophy, muscle contracture, and muscle pain, the mechanisms of which are related to macrophage accumulation. However, muscle contractile exercise through a belt electrode device may mitigate macrophage accumulation. We hypothesized that such exercise would be effective in preventing myofiber atrophy, muscle contracture, and muscular pain. This study tested this hypothesis in immobilized rat gastrocnemius muscle. A total of 32 rats were divided into the following control and experimental groups: immobilization (immobilized treatment only), low-frequency (LF; immobilized treatment and muscle contractile exercise with a 2 s (do) /6 s (rest) duty cycle), and high-frequency (HF; immobilized treatment and muscle contractile exercise with a 2 s (do)/2 s (rest) duty cycle). Electrical stimulation was performed at 50 Hz and 4.7 mA, and muscle contractile exercise was applied to the lower limb muscles for 15 or 20 min/session (once daily) for 2 weeks (6 times/week). After the behavioral tests, the bilateral gastrocnemius muscles were collected for analysis. The number of macrophages, the Atrogin-1 and MuRF-1 mRNA expression, and the hydroxyproline content in the HF group were lower than those in the immobilization and LF groups. The cross-sectional area (CSA) of type IIb myofibers in the superficial region, the PGC-1α mRNA expression, and the range of motion of dorsiflexion in the HF group were significantly higher than those in the immobilization and LF groups. The pressure pain thresholds in the LF and HF groups were significantly higher than that in the immobilization group, and the nerve growth factor (NGF) content in the LF and HF groups was significantly lower than that in the immobilization group. Muscle contractile exercise through the belt electrode device may be effective in preventing immobilization-induced myofiber atrophy, muscle contracture, and muscular pain in the immobilized rat gastrocnemius muscle.

PSX-A-5 Effects of High- and low-Fiber Diets on Growth Performance and Intestinal Oxidative Stress in Growing-Finishing Pigs

Abstract Feed is the most expensive input in commercial pork production. In order to reduce feed costs, using high-fiber ingredients has become a common practice. Moderate levels of fiber can maintain intestinal physiological function and promote intestinal health. Oxidative stress is linked to impaired nutrient absorption and growth performance. This study investigated the effects of high-fiber (HF, 6% crude fiber) and low-fiber (LF, 3% crude fiber) diets on growth performance and intestinal oxidative stress parameters in growing-finishing pigs. Forty growing pigs (27.07 ± 1.26 kg BW) were randomly assigned to 2 treatments with 10 replicates of 2 pigs per pen. Pigs were weighed on days 35, 42 and 70. The feed intake was recorded daily to calculate growth performance parameters. On day 70, eight pigs in each treatment group were randomly selected and euthanized to obtain jejunum to measure oxidative stress status. Pigs fed an HF diet were heavier than those fed an LF diet on days 35, 42 and 70 (P &amp;lt; 0.05). During the whole feeding period, pigs fed an HF diet had higher average daily gain than those fed an LF diet (P &amp;lt; 0.05). The LF diet resulted in increased levels of malondialdehyde (P &amp;lt; 0.05) in the jejunum, suggesting that the LF diet contributed to oxidative stress in the jejunum. The LF diet also led to a significant increase in glutathione and oxidized glutathione levels (P &amp;lt; 0.05) in the jejunum, indicating that pigs fed an LF diet needed to produce more antioxidant substances to cope with oxidative stress in the intestine. This was accompanied by a significant increase in glutamate-cysteine ligase in the jejunum of the LF group (P &amp;lt; 0.05). These results suggest that the HF diet can improve growth performance and maintain intestinal health in growing-finishing pigs by reducing intestinal oxidative stress.

Effects of dietary concentrate to forage ratio on milk performance, milk amino acid composition and milk protein synthesis of lactating donkeys

The purpose of this experiment was to compare the effects of the dietary concentrate-to-forage ratio on lactating donkey milk production performance, milk protein synthesis and milk amino acid composition by replacing part of low-quality roughage with alfalfa. The second objective was to investigate the possible mechanism by which the dietary concentrate-to-forage ratio affects donkey milk protein synthesis. These results will provide a theoretical basis for the rational preparation of feed and optimization of the amino acid composition of donkey milk. This experiment adopted a single-factor randomized trial design. Sixteen Dezhou donkeys with similar ages (6.73 ± 1.16 years), weights (250.84 ± 27.21 kg), parities (3.25 ± 1.12 foals) and lactation days (40 ± 4 days) were selected and evenly (n = 8 per group) divided into a low forage feed group (LF) and a high forage feed group (HF). The LF group was fed a diet with a concentrate to forage feed ratio of 40:60, and the HF group was fed a diet with a concentrate to forage feed ratio of 30:70. The energy and protein levels of the two diets were the same, and the test period lasted 8 weeks. Compared with the LF group, the concentrations of total protein, prolactin and hydrocortisone in the serum of donkeys in the HF group were increased (P < 0.05); the apparent digestibility of dry matter, neutral detergent fiber and energy digestibility and energy metabolic of the donkeys in the HF group were improved (P < 0.05); the concentrations of albumin, epidermal growth factor and urea phosphate tended to decrease (0.05 < P < 0.10); the concentrations of threonine (Thr), lysine (Lys), methionine, histidine, proline and limiting amino acids (LAAs) in the plasma and nitrogen metabolism of the donkeys in the HF group were increased (P < 0.05); milk yield, production and synthesis efficiency of milk protein and most amino acid (AA) production in milk (except serine and valine) were increased (P < 0.05); the proportions of Thr, Lys phenylalanine and LAA in donkey milk were increased (P < 0.05); the final body weight and the body weight gain of donkey foals in the HF group was improved. Under the condition of feeding the same energy and nitrogen levels, increasing the proportion of forage from 60% to 70% by replacing part of low-quality forage with alfalfa promoted the protein and energy metabolism, increased milk protein synthesis efficiency, milk production efficiency and milk production performance, improved the proportion of LAAs and optimized the AA composition of donkey milk.

The Association Between Long-Term Spicy-Food Consumption and the Incidence of Chronic Postsurgical Pain After Cesarean Delivery: An Observational Study.

BackgroundOur previous study found that a long-term diet incorporating spicy foods can reduce the human basal pain threshold. Capsaicin is the pungent ingredient in chili peppers. Transient receptor potential vanilloid type1 is the capsaicin receptor expressed in the oral cavity and is the primary sensory neuron of the “pain” pathway. Few studies have examined the association between long-term spicy diet and chronic postsurgical pain (CPSP). Women who underwent elective cesarean section (eCS) have consistent characteristics of CPSP. This study aimed to investigate the relationship between a long-term spicy diet and the incidence of CPSP after eCS.MethodsParticipants were divided into a low frequency group (LF, numerical rating scale (NRS)<5) for spicy food consumption and a high frequency group (HF, NRS≥5) by receiver operator characteristic analysis. The primary outcome was the incidence of CPSP three months after eCS. Propensity score matching (PSM) analysis was performed between the two frequency groups. Stepwise logistic regression analysis was then performed.ResultsOf the 1029 enrolled patients, data from 982 were analyzed 3 months after eCS. After PSM, the incidence of CPSP in the HF group (30.1% [108/359]) was higher than that in the LF group (19.8% [71/359]; P = 0.001). Compared with the LF group, the risk of CPSP in the HF group increased 1.61 times by 3 months (95% CI 1.18–2.20, P = 0.003). PSM results found that 1 year, the incidence of CPSP in the HF group (15.2% [56/369]) was higher than that in the LF group (8.1% [30/369], P = 0.003).ConclusionWith an NRS≥5 as a boundary, women who consumed spicy food ≥ 2 days/week were more likely to have CPSP than those who consumed spicy food < 2 days/week.

Effects of dietary fishmeal levels on adenosine triphosphate-related compounds and freshness of raw and cooked muscle in large yellow croaker Larimichthys crocea

A comparative study on the adenosine triphosphate (ATP)-related compounds and freshness of raw and cooked muscle in wild and farmed large yellow croaker (Larimichthys crocea) was performed. The two groups of farmed fish (189.87 ± 0.89 g) were fed two diets containing 44% (CF) and 25% (LF) of fish meal (FM) for 82 days, respectively. Results showed that the contents of ATP, adenosine diphosphate (ADP), adenosine monophosphate (AMP) and inosine 5′-monophosphate (IMP) were highest in raw and cooked muscle of wild fish, while these values were lowest in raw and cooked muscle of fish fed LF. Conversely, wild fish had the lowest inosine (Ino) and hypoxanthine (Hx) contents in raw and cooked flesh, while the LF group had the highest levels of Ino and Hx in raw and cooked flesh. Fish fed CF had significantly higher amounts of ATP, ADP and IMP in raw muscle, ATP and AMP in cooked muscle than those in the LF group (P < 0.05). The above results allowed wild fish to have the lowest K and Ki values, closely followed by fish fed CF, and the LF group have the highest values in raw and cooked muscle (P < 0.05). Meanwhile, cooking significantly decreased ATP, ADP and IMP contents, while increased AMP and Ino levels, inducing significantly higher K and Ki values in cooked muscle of all the treatments (P < 0.05). In conclusion, lower dietary FM level could decrease the contents of IMP and its precursors, and increase the degradation products of IMP in muscle, leading to the declines in umami and freshness. The stronger umami of wild large yellow croaker could be related to the higher precursors and lower degradation products of IMP in fillets.

Influence of sire fertility status on conceptus-induced transcriptomic response of the bovine endometrium.

The aim was to examine the effect of sire fertility status on conceptus-induced changes in the bovine endometrial transcriptome. To generate elongated conceptuses, Day 7 blastocysts produced in vitro using frozen-thawed sperm from Holstein Friesian bulls (3 High fertility, HF and 3 Low fertility, LF) were transferred in groups of 5–10 into synchronized heifers (n = 7 heifers per bull) and recovered following slaughter on Day 15. Day 15 endometrial explants recovered from the uterine horn ipsilateral to the corpus luteum were recovered from synchronized cyclic heifers (n = 4). Explants from each heifer were co-cultured for 6 h in RPMI medium alone (Control) or with 100 ng/ml ovine recombinant interferon tau (IFNT) or with a single conceptus from each HF or LF bull. After 6 h, explants were snap frozen and stored at −80°C. Extracted mRNA was subjected to RNA-seq and the resulting data were analyzed with R software. The numbers of differentially expressed genes (DEG; FDR<0.05) were: HF vs. Control: 956; LF vs. Control: 1021; IFNT vs. Control: 1301; HF vs. LF: 2. Unsurprisingly, the majority of DEG (658) were common to all comparisons and were related to IFNT-induced changes in the endometrium. Prior to applying the adjusted p-value, there were 700 DEG between HF and LF, with 191 and 509 genes more expressed in HF or LF, respectively (p < 0.05). Overrepresentation analysis of KEGG pathways (FDR<0.05), revealed that DEG with higher expression in LF were involved in cell cycle and proteolysis, while those upregulated DEG by HF conceptuses were strongly associated with immune process pathways, such as TNF, NF-kappa B, cytokine-cytokine receptor interaction, and TLR signaling. These pathways were also enriched by DEG upregulated by IFNT compared to the Control. Furthermore, only the HF, and not the LF group, affected the expression of most genes in these pathways (p < 0.05) according to a negative binomial regression model. Finally, a weighted gene co-expression network analysis revealed two clusters of co-expressed genes associated with the HF conceptuses (p < 0.05), which were also enriched for the aforementioned pathways. In conclusion, HF conceptuses, similar to IFNT treatment, stimulated multiple pathways involved in immune response, which were apparently not affected by LF conceptuses.

Lactoferrin Prevents Chronic Alcoholic Injury by Regulating Redox Balance and Lipid Metabolism in Female C57BL/6J Mice.

This study aimed to investigate the preventive effects of lactoferrin (Lf) on chronic alcoholic liver injury (ALI) in female mice. Female C57BL/6J mice were randomly divided into four groups: control group (CON), ethanol administration group (EtOH), low-dose Lf treatment group (LLf), and high-dose Lf group (HLf). In the last three groups, chronic ALI was induced by administering 20% ethanol ad libitum for 12 weeks. Mice in the CON and EtOH groups were fed with AIN-93G diet. Meanwhile, 0.4% and 4% casein in the AIN-93G diet were replaced by Lf as the diets of LLf and HLf groups, respectively. HLf significantly reduced hepatic triglyceride content and improved pathological morphology. HLf could inhibit cytochrome P450 2E1 overexpression and promote alcohol dehydrogenase-1 expression. HLf activated protein kinase B and AMP-activated protein kinase (AMPK), as well as upregulating nuclear-factor-erythroid-2-related factor-2 expression to elevate hepatic antioxidative enzyme activities. AMPK activation also benefited hepatic lipid metabolism. Meanwhile, HLf had no obvious beneficial effects on gut microbiota. In summary, Lf could alleviate chronic ALI in female mice, which was associated with redox balance and lipid metabolism regulation.

Integrative Analysis of the Transcriptome and Metabolome Reveals Genes Involved in Phenylpropanoid and Flavonoid Biosynthesis in the Trapa bispinosa Roxb.

BackgroundTrapa bispinosa Roxb. is grown worldwide as an important aquatic cash crop. Current research on Trapa bispinosa primarily focuses on the separation and identification of active ingredients, as well as the inhibitory effect on tumors; however, research on the molecular mechanism of secondary metabolite accumulation is rather limited. Consequently, an integrative analysis of transcriptome and metabolome is required to identify the key metabolic pathways, and key genes, and to explain the molecular mechanism of Trapa bispinosa.ResultsThe biosynthesis pathways of phenolics in Trapa bispinosa were examined through transcriptome and metabolome analyses. Transcriptome analysis yielded 42.76 million clean reads representing 81,417 unigenes with an average length of 1,752 bp. KEGG pathway analysis revealed that 1,623 unigenes, including 88 candidate unigenes related to phenolics biosynthesis, were up-regulated in Trapa bispinosa shell (FR) when compared to leaves (LF), root (RT), and stem (ST). The FR vs. LF group had the highest number of specific genes involved in phenylpropanoid, flavonoid, flavone, and flavonol biosynthesis pathways compared to all other comparison groups. In addition, RNA sequencing revealed 18,709 SSRs spanning 14,820 unigenes and 4,387 unigenes encoding transcription factors. Metabolome analysis identified 793 metabolites, including 136 flavonoids and 31 phenylpropane compounds. In the FR group compared to the LF group, there were 202 differentially accumulated metabolites (DAMs). The combined transcriptome and metabolome analyses indicated a significant correlation between 1,050 differentially expressed genes (DEGs) and 62 DAMs. This view proposes a schematic of flavonoid biosynthesis in the FR vs. LF group, providing evidence for the differences in genes and metabolites between FR and LF.ConclusionIn this study, through de novo transcriptome assembly and metabolome analysis, several DEGs and DAMs were identified, which were subsequently used to build flavonoid biosynthesis pathways and a correlation network. The findings pave the way for future research into the molecular mechanisms and functional characterization of Trapa bispinosa candidate genes for phenolics biosynthesis.

Comparison of surface roughness of root cementum and orthodontically induced root resorption craters from high- and low-fluoridation areas: a 3D confocal microscopy study

BackgroundFluoride has a major role in strengthening the structure of enamel against acids. Despite differences between caries and root resorption processes, both events inherently involve acidic dissolution of dental tissues. The aim of the present study was to investigate the effects of water fluoridation levels on the surface roughness of root cementum and resorption craters. The findings provided more insight into the influence of fluoride on the surfaces of intact cementum surface and resorption craters.MethodsTwenty-eight orthodontic patients were recruited from two cities in Turkey, with high (≥ 2 ppm) and low (≤ 0.05 ppm) water fluoridation. These patients needed bilateral maxillary first premolar extraction as part of their orthodontic treatment and were allocated into two study groups (n = 14 in each group) based on water fluoridation exposure level: the high-fluoride group (HF) and low-fluoride group (LF). 150 g of buccal tipping forces was applied to all maxillary first premolar teeth for 12 weeks with a beta-titanium spring which was reactivated every 4 weeks. All maxillary premolars were removed at the end of the experiment for surface roughness assessment using three-dimensional confocal microscopy and the associated software. The buccal root surface and the largest buccal resorption crater were investigated.ResultsResorption craters were significantly rougher in LF group compared to HF group (p = 0.002). Craters were rougher than the intact root surfaces (p = 0.000). Cervical and apical regions were significantly rougher than the middle region (p = 0.000 and p = 0.024, respectively).ConclusionsHigher water fluoridation level of ≥ 2 ppm resulted in significantly smoother root resorption craters than low water fluoridation level of ≤ 0.05 ppm when the teeth were subjected to 150 g of buccal tipping force. Fluoride seems to have a protective role at the interface of root resorption, and further mineral or histological studies may shed light on the exact protective process against root resorption.

Implementation of a CT-derived correction factor to refine the measurement of aortic valve area and stroke volume using Doppler echocardiography improves grading of severity and prediction of prognosis in patients with severe aortic stenosis

AimsTo assess rates of reclassification of severity and associated 5-year survival in patients with severe aortic stenosis (AS) and preserved left ventricular ejection fraction (LVEF) after application of a CT-derived correction factor (CF) to refine the measurement of aortic valve area (AVA) and stroke volume index (SVi) using Doppler echocardiography. Methods and resultsWe enrolled 1450 patients with severe AS and preserved LVEF from a French registry. Multiplication of echocardiographic LV outflow tract diameter by a CT-derived CF of 1.13 to calculate the AVA and SVi using the continuity equation resulted in reclassification of 39% of patients from severe to moderate AS (AVA > 1 cm2) and 77% from low flow (LF, SVi < 35 ml/m2) to normal flow (NF, SVi ≥ 35 ml/m2). After application of the CF, 5-year survival with conservative management was 50 ± 4% for severe AS compared to 62 ± 4% for moderate AS (p < 0.001). A strategy of medical management followed by intervention for severe AS was associated with higher risk of mortality over 5-year follow-up after adjustment for covariates and application of the CF (HR 1.35 [1.10–1.55], p = 0.015). Five-year survival was also poorer in patients remaining in the LF group after application of the CF, even after valve intervention (72%, 66% and 47% for NF to NF, LF to NF and LF to LF, respectively). After adjustment for covariates (including intervention), risk of mortality was higher in LF to LF patients compared to NF to NF (HR 1.78 [1.25–2.56]), but similar for NF to NF and LF to NF (HR 1.20 [0.90–1.60]). ConclusionRefined accuracy of echocardiographic LV outflow tract diameter measurement using a CF of 1.13 before derivation of AVA and SVi in patients with severe AS and preserved LVEF allows improved grading of severity, and prediction of prognosis. We recommend implementation of the CF during routine echocardiography when using the continuity equation for Doppler haemodynamic measurements.

Intratumoral fibrosis and patterns of immune infiltration in clear cell renal cell carcinoma

BackgroundIntratumoral fibrosis was positively correlated with histological grade of renal clear cell carcinoma (ccRCC) and intratumoral inflammation. However, the association of intratumoral fibrosis with the immune infiltration of ccRCC was few evaluated.MethodsWe used the second harmonic generation (SHG)-based imaging technology and evaluated the intratumoral fibrosis in ccRCC, and then divided the patients into the high fibrosis group (HF) and the low fibrosis group (LF). Meanwhile, the Kaplan–Meier survival curve analysis was performed to analyze the relationship between intratumoral fibrosis and the disease-free survival rate. Antibody arrays were used for seeking difference in cytokines and immune infiltration between the HF group (N = 11) and LF group (N = 11). The selected immune infiltration marker was then verified by immunohistochemistry (IHC) staining in 45 ccRCC samples.ResultsOut of 640 cytokines and immune infiltration markers, we identified 115 proteins that were significantly different in quantity between ccRCC and adjacent normal tissues. In addition, the Venn diagram indicated that six proteins, including Cytotoxic T-Lymphocyte Associated Protein 4 (CTLA4), were significantly associated with intratumoral fibrosis (p < 0.05). The GO/KEGG enrichment analysis indicated that the proteins associated with intratumoral fibrosis were involved in the immunity and tumor-infiltrating lymphocytes. The expression of the CTLA4 was negatively correlated with collagen level, confirmed by IHC staining of CTLA4 (p < 0.05).ConclusionsThe study indicated that the intratumoral fibrosis level was negatively correlated with the expression of CTLA4 in the tumor immune microenvironment of the ccRCC, which posed the potential value of targeting the stroma of the tumor, a supplement to immunotherapy. However, the specific mechanism of this association is still unclear and needs further investigation.

Dietary Micronutrients Reduce Insulin Resistance via Adipose Tissue Modulation in Mice Fed a High Fat Diet

ObjectivesObesity is associated with increased insulin resistance (IR) and white adipose tissue (WAT) dysregulation (e.g., decrease in PPAR-γ mRNA expression and impaired leptin sensitivity). Dietary vitamins A, B1, B6, and B12, selenium (Se) and zinc (Zn) have been shown to reduce IR in obesity in animals and humans, but mechanisms have not been defined. The objective of this study is to investigate the effects of selected micronutrients on adipose tissue metabolism and IR in a mouse model of diet-induced obesity (DIO). We hypothesize that these additions modulate genes regulating IR in WAT and improve leptin sensitivity.Methods28 DIO male mice were randomly assigned to two high fat (HF, 60 kcal % fat) diets with or without the inclusion of a mineral-vitamin mixture (MVM: 5 × vitamin A, B1, B6, B12, zinc, and 2 × selenium), respectively. Similarly, 28 lean mice were randomized into a low-fat diet (LF, 10 kcal % fat) with or without the MVM. Mice were fed diets ad libitum for 8 weeks. Bodyweight (BW), IR, serum glucose, fasting insulin, C-peptide, leptin, as well as mRNA expression of genes involved in insulin signaling and adipokine secretion were measured in epididymal white adipose tissue (eWAT).ResultsCompared to HF mice, HF-MVM mice exhibited reduced body weight gain over time (by 6%, P < 0.05), improved insulin sensitivity (P < 0.05), reduced fasting glucose (by 18%), insulin (by 45%), C-peptide concentrations (by 26%) and Homeostatic Model Assessment for Insulin Resistance (by 47%, HOMA-IR, P < 0.05 for all). Similarly, LF-MVM had reduced fasting glucose level (P < 0.05) compared to LF mice. As expected, serum leptin concentration (adjusted for a visceral fat mass) was 6-fold higher in HF mice compared to LF mice; HF-MVM mice had lower leptin level compared to the HF mice suggesting improved leptin sensitivity. Ppar-γ gene expression in eWAT was 77% lower in HF vs LF group, suggesting eWAT dysfunction and systemic IR in DIO mice; the addition of MVM to LF diet attenuated this effect.ConclusionsThis MVM mixture may reduce IR through the upregulation of the PPAR-γ system in WAT and improve leptin sensitivity in DIO. Understanding the mechanism of action of micronutrients in reducing IR in a highly relevant animal model will provide new avenues for identifying food component solutions to modify IR in humans.Funding SourcesSupport provided by Various Sponsors (unrestricted funds).

Regulation of Energy Metabolism by Dietary pH and Protein Source in Diet‐induced Obese Mice and Female Mice

Consumption of foods with acidic pH has been proposed as a potential contributor to negative metabolic health outcomes. Thus, in this work, we determined the main effects of dietary pH during onset of obesity in male and female B6 mice. We hypothesized that metabolic health will be improved by consuming a diet containing pH‐enhanced ground beef or casein diets, compared to a non‐pH‐enhanced diets. To test this hypothesis, we designed a mouse study using the following 8 groups: Low‐fat diet + casein protein (LFC), LF + casein with pH enhanced/ammonia (LFCN), LF + beef protein (LFB), LFB with pH enhanced (LFBN), High‐fat diet + casein protein (HFC), HF + casein with pH enhanced (HFCN), HF + beef protein (HFB) and HFB with pH enhanced (HFBN). Mice were fed these diets for 12 weeks, during which we measured changes in body weight and food intake. Mice were individually placed in metabolic cages (TSE PhenoMaster System) at ambient temperature and a 12‐hour light/dark cycle at week 11. The relationship between metabolic rate and body mass was normalized, and we evaluated energy balance (H), respiratory exchange ratio (RER), VCO2, VO2, feed, drink and distance traveled by the mice. These metabolic data were analyzed in both day and night periods. Blood and tissues were collected at the end of the study and fat pad weights were measured. Results showed that in males, body weight was increased in all HF groups, regardless of the protein source, while females did not exhibit any differences across groups. However, for energy metabolism, overall, females exhibited the main changes, especially for the interaction between LF and HF groups. These findings suggest that despite the lack of significant weight gain in females, they used fat as energy source based on their RER, VCO2 and VO2 data. During the day, only HF females had decreased VCO2, and at night both male and female exhibited the same pattern of decreased VCO2. During the night, only males from HF groups had reduced VO2. Regarding energy intake, only HF females had increased food consumption compared to males. The main effects of RER in both males and females were related to diet (LF x HF), in both day and night periods. Interestingly, females also presented differences for the interactions of the protein content (casein vs beef) during the day (p&lt;0.0115). The latter suggests that in females, the protein source may be related to energy consumption and substrate utilization. Fat pad weights were increased in males from all HF groups compared to all LF groups. However, in females, only HFC groups had higher fat pad weights compared to LFC. The reduced RER, observed in both male and female mice from HF groups indicates that fat was oxidized and used as the main energy source in these groups, consistent with higher dietary fat content. Taken together, our data suggest that metabolic changes and substrate utilization are affected by dietary fat content (HF) and are more pronounced in female mice. Further analyses in metabolically active tissues from these mice will help elucidate the mechanisms mediating sex‐dependent differences in metabolic effects of dietary pH enhancement, fat content and source of protein.

Fitness Levels and Gender Are Related With the Response of Plasma Adipokines and Inflammatory Cytokines in Prepubertal Children.

Background and AimChanges in adipokines have been related with the development of metabolic syndrome, frequently associated with obesity, and other risk factors. Fitness seems to promote a healthy cardiovascular status and could be a protector factor, just from childhood. Therefore, the present study aimed to evaluate the relationship between fitness levels with plasma adipokines and inflammatory biomarkers in prepubertal children.MethodsOne hundred and thirty-seven healthy normal-weight prepubertal children were recruited from local schools and divided after performing the fitness tests, into two groups according to fitness level—low cardiovascular fitness group (LF) and equal or higher cardiovascular fitness group (HF). Anthropometric variables, blood pressure (BP) and plasma insulin, and leptin, resistin, adiponectin, tumor necrosis factor-alpha, hepatic growth factor, interleukin (IL)-8, monocyte chemoattractant protein-1, nerve growth factor (NGF), and plasminogen activator inhibitor-1 (PAI-1) were measured fasting in both groups to be compared. Univariate analysis of variance, comparative analysis, binary logistic regression, stepwise linear regression, and principal component analysis were conducted to evaluate the association between fitness, BMI, gender, and the biochemical parameters.ResultsGirls and boys with HF presented lower waist circumference Z-score, BMI Z-score, systolic BP (only boys) as well as lower levels of leptin and NGF compared with their respective LF group. Regarding the association between variables, fitness showed an inverse relationship with BMI Z-score, leptin, PAI-1, HOMA-IR, resistin, IL-8, and NGF.ConclusionAn adequate level of fitness seems to protect against risk factors related to low-grade inflammation and altered adipokines that are related to the onset of obesity just from the prepubertal stage.

The Frequency Effect of the Motor Imagery Brain Computer Interface Training on Cortical Response in Healthy Subjects: A Randomized Clinical Trial of Functional Near-Infrared Spectroscopy Study.

BackgroundThe motor imagery brain computer interface (MI-BCI) is now available in a commercial product for clinical rehabilitation. However, MI-BCI is still a relatively new technology for commercial rehabilitation application and there is limited prior work on the frequency effect. The MI-BCI has become a commercial product for clinical neurological rehabilitation, such as rehabilitation for upper limb motor dysfunction after stroke. However, the formulation of clinical rehabilitation programs for MI-BCI is lack of scientific and standardized guidance, especially limited prior work on the frequency effect. Therefore, this study aims at clarifying how frequency effects on MI-BCI training for the plasticity of the central nervous system.MethodsSixteen young healthy subjects (aged 22.94 ± 3.86 years) were enrolled in this randomized clinical trial study. Subjects were randomly assigned to a high frequency group (HF group) and low frequency group (LF group). The HF group performed MI-BCI training once per day while the LF group performed once every other day. All subjects performed 10 sessions of MI-BCI training. functional near-infrared spectroscopy (fNIRS) measurement, Wolf Motor Function Test (WMFT) and brain computer interface (BCI) performance were assessed at baseline, mid-assessment (after completion of five BCI training sessions), and post-assessment (after completion of 10 BCI training sessions).ResultsThe results from the two-way ANOVA of beta values indicated that GROUP, TIME, and GROUP × TIME interaction of the right primary sensorimotor cortex had significant main effects [GROUP: F(1,14) = 7.251, P = 0.010; TIME: F(2,13) = 3.317, P = 0.046; GROUP × TIME: F(2,13) = 5.676, P = 0.007]. The degree of activation was affected by training frequency, evaluation time point and interaction. The activation of left primary sensory motor cortex was also affected by group (frequency) (P = 0.003). Moreover, the TIME variable was only significantly different in the HF group, in which the beta value of the mid-assessment was higher than that of both the baseline assessment (P = 0.027) and post-assessment (P = 0.001), respectively. Nevertheless, there was no significant difference in the results of WMFT between HF group and LF group.ConclusionThe major results showed that more cortical activation and better BCI performance were found in the HF group relative to the LF group. Moreover, the within-group results also showed more cortical activation after five sessions of BCI training and better BCI performance after 10 sessions in the HF group, but no similar effects were found in the LF group. This pilot study provided an essential reference for the formulation of clinical programs for MI-BCI training in improvement for upper limb dysfunction.

Effects of corticospinal tract integrity on upper limb motor function recovery in stroke patients treated with repetitive transcranial magnetic stimulation.

The bimodal balance-recovery model predicts that corticospinal tract (CST) integrity in the affected hemisphere influences the partterns of brain recovery after stroke. Repetitive transcranial magnetic stimulation (rTMS) has been used to promote functional recovery of stroke patients by modulating motor cortical excitability and inducing reorganization of neural networks. This study aimed to explore how to optimize the efficiency of repetitive transcranial magnetic stimulation to promote upper limb functional recovery after stroke according to bimodal balance-recovery model. 60 patients who met the inclusion criteria were enrolled to high CST integrity group (n = 30) or low CST integrity group (n = 30), and further assigned randomly to receive high-frequency rTMS (HF-rTMS), low-frequency rTMS (LF-rTMS) or sham rTMS in addition to routine rehabilitation, with 10 patients in each group. Outcome measures included Fugl-Meyer scale for upper extremity (FMA-UE), Wolf Motor Function (WMFT) scale and Modified Barthel Index (MBI) scale which were evaluated at baseline and after 21 days of treatment. For patients with high CST integrity, the LF group achieved higher FMA-UE, WMFT and MBI scores improvements after treatment when compared to the HF group and sham group. For patients with low CST integrity, after 21 days treatment, only the HF group showed significant improvements in FMA-UE and WMFT scores. For MBI assessment, the HF group revealed significantly better improvements than the LF group and sham group. For stroke patients with high CST integrity, low-frequency rTMS is superior to high-frequency rTMS in promoting upper limb motor function recovery. However, only high-frequency rTMS can improve upper limb motor function of stroke patients with low CST integrity.