Development of Hsp90 inhibitor to regulate cytokine storms in excessive delayed- and acute inflammation.

BACKGROUND: The surplus cytokines remaining after use in the early stages of the inflammatory response stimulate immune cells even after the response is over, causing a secondary inflammatory response and ultimately damaging the host, which is called a cytokine storm. Inhibiting heat shock protein 90 (Hsp90), which has recently been shown to play an important role in regulating inflammation in various cell types, may help control excessive inflammatory responses and cytokine storms. METHODS: We discovered an anti-inflammatory compound by measuring the inhibitory effect of CD86 expression on spleen DCs (sDCs) using the chemical compounds library of Hsp90 inhibitors. Subsequently, to select the hit compound, the production of cytokines and expression of surface molecules were measured on the bone marrow-derived DCs (BMDCs) and peritoneal macrophages. Then, we analyzed the response of antigen-specific Th1 cells. Finally, we confirmed the effect of the compound using acute lung injury (ALI) and delayed-type hypersensitivity (DTH) models. RESULTS: We identified Be01 as the hit compound, which reduced CD86 expression the most in sDCs. Treatment with Be01 decreased the production of pro-inflammatory cytokines (IL-6, TNF-α, and IL-1ß) in BMDC and peritoneal macrophages stimulated by LPS. Under the DTH model, Be01 treatment reduced ear swelling and pro-inflammatory cytokines in the spleen. Similarly, Be01 treatment in the ALI model decreased neutrophil infiltration and lower levels of secreted cytokines (IL-6, TNF-α). CONCLUSIONS: Reduction of CD80 and CD86 expression on DCs by Be01 indicates reduced secondary inflammatory response by Th1 cells, and reduced release of pro-inflammatory cytokines by peritoneal macrophages may initially control the cytokine storm.

Kapok fiber composites minimizing secondary waste and disposal costs for large-scale radioactive liquid treatment.

Conventional liquid treatments for large-scale, low-level radioactive wastewater, such as ion exchange and waste solidification, face challenges due to the large amounts of secondary waste and high disposal costs. A new large-scale decontamination method is proposed that uses kapok fiber composites for rapid radionuclide adsorption and high volume reduction to minimize secondary waste. The composite consists of natural zeolite and kapok holocellulose, which has high water-soaking ability and low-temperature pyrolysis. The kapok composites, fabricated using a commercial wet-laid nonwoven manufacturing process, absorbs 99% of low-level radioactive cesium in 20 min, reducing the volume by 98% and the weight by 47% at 300 °C. The low-temperature pyrolysis process below 300 °C prevents cesium desorption and gasification by avoiding zeolite destruction. The mass-producible kapok composites can be used for adsorbing various radionuclides in large-scale wastewater by attaching specific adsorbents for target isotopes to the composites.

Targeting Heme Oxygenase 2 (HO2) with TiNIR, a Theragnostic Approach for Managing Metastatic Non-Small Cell Lung Cancer.

Despite notable advancements in cancer therapeutics, metastasis remains a primary obstacle impeding a successful prognosis. Our prior study has identified heme oxygenase 2 (HO2) as a promising therapeutic biomarker for the aggressive subsets within tumor. This study aims to systematically evaluate HO2 as a therapeutic target of cancer, with a specific emphasis on its efficacy in addressing cancer metastasis. Through targeted inhibition of HO2 by TiNIR (tumor-initiating cell probe with near infrared), we observed a marked increase in reactive oxygen species. This, in turn, orchestrated the modulation of AKT and cJUN activation, culminating in a substantial attenuation of both proliferation and migration within a metastatic cancer cell model. Furthermore, in a mouse model, clear inhibition of cancer metastasis was unequivocally demonstrated with an HO2 inhibitor administration. These findings underscore the therapeutic promise of targeting HO2 as a strategic intervention to impede cancer metastasis, enhancing the effectiveness of cancer treatments.

Anti-fibrotic and anti-stricture effects of biodegradable biliary stents braided with dexamethasone-impregnated sheath/core structured monofilaments.

Endoscopic biliary stent insertion has been widely used for the treatment of benign biliary stricture (BBS). Thus, the development of stent materials in the perspectives of structure, mechanical properties, and biocompatibility has been also studied. However, conventional metal and plastic stents have several disadvantages, such as repeated procedures to remove or exchange them, dislodgment, restenosis, biocompatibility, and poor mechanical properties. Sustainable effectiveness, attenuation and prevention of fibrosis, and biocompatibility are key factors for the clinical application of stents to BBS treatment. In addition, loading drugs could show synergistic effects with stents' own performance. We developed a dexamethasone-eluting biodegradable stent (DBS) consisting of a sheath/core structure with outstanding mechanical properties and sustained release of dexamethasone, which maintained its functions in a BBS duct over 12 weeks in a swine model. The insertion of our DBS not only expanded BBS areas but also healed secondary ulcers as a result of the attenuation of fibrosis. After 16 weeks from the insertion, BBS areas were totally improved, and the DBS was degraded and thoroughly disappeared without re-intervention for stent removal. Our DBS would be an effective clinical tool for non-vascular diseases. STATEMENT OF SIGNIFICANCE: This study describes the insertion of a drug-eluting biodegradable stent (DBS) into the bile duct. The sheath/core structure of DBS confers substantial durability and a sustained drug release profile. Drug released from the DBS exhibited anti-fibrotic effects without inflammatory responses in both in vitro and in vivo experiments. The DBS maintained its function over 12 weeks after insertion into the common bile duct, expanding benign biliary stricture (BBS) and reducing inflammation to heal secondary ulcers in a swine BBS model. After 16 weeks from the DBS insertion, the DBS thoroughly disappeared without re-intervention for stent removal, resulting in totally improved BBS areas. Our findings not only spotlight the understanding of the sheath/core structure of the biodegradable stent, but also pave the way for the further application for non-vascular diseases.

Life-threatening Gastrointestinal Bleeding from a Dieulafoy's Lesion in the Duodenum: A Case Report.

Dieulafoy's lesion is a rare cause of gastrointestinal bleeding, accounting for approximately 1-2% of all cases of gastrointestinal bleeding. Dieulafoy's lesion usually occurs in the lesser curvature of the stomach within six centimeters of the gastroesophageal junction. On the other hand, extragastric Dieulafoy's lesions are uncommon. Diagnosing an extragastric Dieulafoy's lesion by endoscopy can be challenging because of its small size and obscure location. The key elements for an accurate diagnosis include heightened awareness and a careful early endoscopic evaluation following a bleeding episode. Various endoscopic hemostatic techniques can be used for treatment. This paper presents a case of successful hemostasis using argon plasma coagulation for a life-threatening duodenal Dieulafoy's lesion.

Exercise enhances placental labyrinth trophoblast development by activation of PGC-1α and FNDC5/irisin†.

Placental chorion/labyrinth trophoblasts are energy demanding which is met by the mitochondrial oxidative phosphorylation. Exercise enhances placental development and mitochondrial biogenesis, but the underlying mechanisms remain poorly understood. To address, female C57BL/6 J mice were randomly assigned into two groups: a control group and an exercise (EX) group. All animals were acclimated to treadmill exercise for 1 week before mating, but only the EX group was subjected to daily exercise during pregnancy from embryonic day (E) 1.5 to E16.5. Placenta were collected at E18.5 for biochemical and histochemical analyses, and primary trophoblast cells were isolated from the E18.5 placenta for further analyses. The data showed that exercise during pregnancy promoted the expression of syncytiotrophoblast cell markers, indicating trophoblast cell differentiation, which was closely associated with elevated mitochondrial biogenesis and oxidative metabolism in the E18.5 placenta. In addition, exercise during pregnancy activated peroxisome proliferator-activated receptor γ coactivator-1 α (PGC-1α), which was associated with upregulated placental α-ketoglutarate and the expression of isocitrate dehydrogenases and ten-eleven translocations, facilitating DNA demethylation of the Pgc1a promoter. Furthermore, exercise upregulated fibronectin type III domain containing 5 expression and the secretion of its cleaved form, irisin, which is known to activate PGC-1α. These data suggest that exercise-induced activation of PGC-1α, via epigenetic modifications, is responsible for promoting mitochondrial energy metabolism and chorion/labyrinth trophoblast development.

Effect of Mixture of Recombinant Human Bone Morphogenic Protein-2 and demineralized Bone Matrix in Lateral Lumbar Interbody Fusion.

Objective: This study aims to determine the optimal dose of recombinant-human Bone Morphogenic Protein-2 (rhBMP-2) for successful bone fusion in minimally invasive Lateral Lumbar Interbody Fusion (MIS LLIF). Previous studies show that rh-BMP is an effective alternative to autologous iliac crest bone graft, but the optimal dose remains uncertain. The study analyzes the fusion rates associated with different rh-BMP doses to provide a recommendation for the optimal dose in MIS LLIF. Methods: 93 patients underwent MIS LLIF using demineralized bone matrix (DBM) or a mixture of rhBMP-2 and DBM as fusion material. The group was divided into the following three groups according to the rhBMP-2 usage. Group A (only DBM was used, n: 27). Group B (1mg of rhBMP-2 per 5cc of DBM paste, n: 41). Group C (2mg of rhBMP-2 per 5cc of DBM paste, n: 25). Demographic data, clinical outcomes, postoperative complication and fusion were assessed. Results: At 12 months post-surgery, the overall fusion rate was 92.3% according to Bridwell fusion grading system. Group B and C, who received rhBMP-2, had significantly higher fusion rates than group A, who received only DBM. However, there was no significant increase in fusion rate when the rhBMP-2 dosage was increased from group B to group C. The group B and C showed significant improvement in back pain and ODI compared to the group A. The incidence of screw loosening was decreased in group B and C, but there was no significant difference in the occurrence of other complications. Conclusion: Usage of rhBMP-2 in LLIF surgery leads to early and increased final fusion rates, which can result in faster pain relief and return to daily activities for patients. The benefits of using rhBMP-2 were not significantly different between the groups that received 1mg/5cc and 2mg/5cc of rhBMP-2. Therefore, it is recommended to use 1mg of rhBMP-2 with 5cc of DBM, taking both economic and clinical aspects into consideration.

The role of infectious disease consultations in the management of patients with fever in a long-term care facility.

BACKGROUND: Infectious disease (ID) clinicians can provide essential services for febrile patients in tertiary hospitals. The aim of this study was to evaluate the role of ID consultations (IDC) in managing hospitalized patients with infections in an oriental medical hospital (OMH), which serves as a long-term care facility. To our knowledge, this is the first study on the role of IDCs in managing patients in an OMH. METHODS: This retrospective study was conducted in an OMH in Seoul, Korea, from June 2006 to June 2013. RESULTS: Among the 465 cases of hospital-acquired fever, 141 (30.3%) were referred for ID. The most common cause of fever was infection in both groups. The peak body temperature of the patient was higher in IDC group (38.8±0.6°C vs. 38.6±0.5°C, p<0.001). Crude mortality at 30 days (14.6% vs. 7.8%, p = 0.043) and infection-attributable mortality (15.3% vs. 6.7%, p = 0.039) were higher in the No-IDC group. Multivariable analysis showed that infection as the focus of fever (adjusted Odd ratio [aOR] 3.49, 95% confidence interval (CI) 1.64-7.44), underlying cancer (aOR 10.32, 95% CI 4.34-24.51,), and multiorgan dysfunction syndrome (aOR 15.68, 95% CI 2.06-119.08) were associated with increased 30-day mortality. Multivariate analysis showed that in patients with infectious fever, appropriate antibiotic therapy (aOR 0.19, 95% CI 0.05-0.76) was the only factor associated with decreased infection-attributable mortality while underlying cancer (aOR 7.80, 95% CI 2.555-23.807) and severe sepsis or septic shock at the onset of fever (aOR 10.15, 95% CI 1.00-102.85) were associated with increased infection-attributable mortality. CONCLUSION: Infection was the most common cause of fever in patients hospitalized for OMH. Infection as the focus of fever, underlying cancer, and MODS was associated with increased 30-day mortality in patients with nosocomial fever. Appropriate antibiotic therapy was associated with decreased infection-attributable mortality in patients with infectious fever.

Clinical characteristics of native vertebral osteomyelitis in patients with history of acupuncture.

BACKGROUND: Considering that vertebral osteomyelitis (VO) can occur via various routes, it can be predicted that clinical characteristics may vary depending on the route of infection or risk factors of the disease. In this study, differences in clinical characteristics, causative pathogens, clinical features and prognosis were investigated in patients of native vertebral osteomyelitis with history of acupuncture. METHODS: This retrospective study was conducted at Kyung Hee University Hospital at Gangdong, Seoul. We extracted data of patients diagnosed with VO from May 2006 to February 2021 using an electronic database. Data on demography, clinical presentation, treatment, causative organisms and clinical outcomes were identified and compared according to the history of acupuncture. RESULTS: A total of 100 patients with VO were reviewed, among which 34 patients had a history of acupuncture prior to the diagnosis of VO. The frequency of Gram-positive cocci (GPC) was significantly higher in the acupuncture group than in the non-acupuncture group (p = 0.016). Abscess was observed more frequently in the acupuncture group than in the non-acupuncture group (p = 0.01). There was no difference in neurological sequelae and recurrence between the two groups. There was no difference in mortality between the two groups. (p = 0.098) CONCLUSION: This study suggests that confirming a history of acupuncture may help predict the pathogen or clinical characteristics of the disease. If the patient has a history of acupuncture, GPC can be considered as the causative organism, and the findings that abscesses and surgical treatment are more common may be helpful in evaluating patients.

Immunoglobulin G4-related Disease of the Small Bowel: A Case of Long-term Remission Achieved by Surgical Resection without Maintenance Therapy.

Immunoglobulin G4-related disease (IgG4-RD) is an immune-mediated fibroinflammatory disease. IgG4-RD can affect any organ system, including the pancreas, bile ducts, salivary glands, mesentery, and retroperitoneum. On the other hand, small intestine involvement is extremely rare. This paper describes a case of IgG4-RD involving the small bowel, particularly at the distal ileum. An 81-year-old female was admitted to the authors' hospital complaining of abdominal pain, dyspepsia, and hematochezia. The laboratory tests, including tumor markers and IgG4, were within normal limits. A colonoscopy did not show any abnormal findings. Abdominal computed tomography revealed segmental aneurysmal dilatation and wall thickening at the distal ileum, suggesting malignant conditions, such as small bowel lymphoma. The patient underwent an exploratory laparoscopy and ileocecectomy to differentiate a malignancy. A histopathology examination revealed dense lymphoplasmacytic infiltration, storiform fibrosis, and IgG4-positive plasma cells (>50 per high power field). The patient was finally diagnosed with IgG4-RD. The patient was followed up in the outpatient clinic for five years without recurrence. This paper suggests that a radical resection without maintenance therapy can be a treatment option, particularly when the IgG4-RD manifests as a localized gastrointestinal tract lesion.

Exercise improves homeostasis of the intestinal epithelium by activation of apelin receptor-AMP-activated protein kinase signalling.

Intestinal remodelling is dynamically regulated by energy metabolism. Exercise is beneficial for gut health, but the specific mechanisms remain poorly understood. Intestine-specific apelin receptor (APJ) knockdown (KD) and wild-type male mice were randomly divided into two subgroups, with/without exercise, to obtain four groups: WT, WT with exercise, APJ KD and APJ KD with exercise. Animals in the exercise groups were subjected to daily treadmill exercise for 3 weeks. Duodenum was collected at 48 h after the last bout of exercise. AMP-activated protein kinase (AMPK) α1 KD and wild-type mice were also utilized for investigating the mediatory role of AMPK on exercise-induced duodenal epithelial development. AMPK and peroxisome proliferator-activated receptor γ coactivator-1 α were upregulated by exercise via APJ activation in the intestinal duodenum. Correspondingly, exercise induced permissive histone modifications in the PR domain containing 16 (PRDM16) promoter to activate its expression, which was dependent on APJ activation. In agreement, exercise elevated the expression of mitochondrial oxidative markers. The expression of intestinal epithelial markers was downregulated due to AMPK deficiency, and AMPK signalling facilitated epithelial renewal. These data demonstrate that exercise-induced activation of the APJ-AMPK axis facilitates the homeostasis of the intestinal duodenal epithelium. KEY POINTS: Apelin receptor (APJ) signalling is required for improved epithelial homeostasis of the small intestine in response to exercise. Exercise intervention activates PRDM16 through inducing histone modifications, enhanced mitochondrial biogenesis and fatty acid metabolism in duodenum. The morphological development of duodenal villus and crypt is enhanced by the muscle-derived exerkine apelin through the APJ-AMP-activated protein kinase axis.

[Recent Updates on Management and Follow-up of Gallbladder Polyps].

Gallbladder polyps are a common incidental finding. Although most of these are benign, differentiating non-neoplastic from neoplastic polyps is challenging. Trans-abdominal ultrasound is the primary imaging study for diagnosing and monitoring gallbladder polyps. In challenging cases, the use of endoscopic ultrasound or contrast-enhanced endoscopic ultrasound could assist in making decisions. According to current guidelines, a cholecystectomy is recommended in patients with polyps measuring 10 mm or larger and in symptomatic patients with polyps measuring less than 10 mm. A cholecystectomy is also recommended if one or more risk factors for malignancy are present in patients with polyps measuring 6-9 mm. These risk factors include age older than 60 years, primary sclerosing cholangitis, Asian ethnicity, and sessile polyps, including focal gallbladder wall thickening >4 mm. Follow-up ultrasound is recommended at six months, one year, and two years for polyps measuring 6-9 mm in patients without risk factors for malignancy, and for polyps less than 5 mm in patients with one or more risk factors for malignancy. Discontinuing the surveillance could be considered in the absence of growth. Follow-up is not required for polyps measuring less than 5 mm in patients without the risk factors for a malignancy. On the other hand, the evidence for the guidelines is still lacking and of low quality. The management of gallbladder polyps should be individualized based on the currently available guidelines.

Ginsenoside Rg3-enriched Korean Red Ginseng extract attenuates Non-Alcoholic Fatty Liver Disease by way of suppressed VCAM-1 expression in liver sinusoidal endothelium.

Background: The incidence and clinical importance of nonalcoholic fatty liver disease (NAFLD) has emerged. However, effective therapeutic strategies for NAFLD have yet to be found. Panax ginseng (P. ginseng) is a traditional herb in Eastern Asia with therapeutic effects in many chronic disorders. However, the precise effects of ginseng extract on NAFLD are currently unknown. In present study, the therapeutic effects of Rg3-enriched red ginseng extract (Rg3-RGE) on the progression of NAFLD were explored. Methods: Twelve-week-old C57BL/6 male mice were fed a chow or western diet supplemented with high sugar water solution with or without Rg3-RGE. Histopathology, immunohistochemistry, immunofluorescence, serum biochemistry, western blot analysis, and quantitative RT-PCR were used for in vivo experiment. Conditionally immortalized human glomerular endothelial cell (CiGEnC) and primary liver sinusoidal endothelial cells (LSECs) were used for in vitro experiments. Results: Eight weeks of Rg3-RGE treatment significantly attenuated the inflammatory lesions of NAFLD. Furthermore, Rg3-RGE inhibited the inflammatory infiltrate in liver parenchyma and the expression of adhesive molecules to LSECs. Moreover, the Rg3-RGE exhibited similar patterns on the in vitro assays. Conclusion: The results demonstrate that Rg3-RGE treatment ameliorates NAFLD progression by inhibiting chemotaxis activities in LSECs.

Flavokawain B Inhibits Growth of Cholangiocarcinoma Cells by Suppressing the Akt Pathway.

BACKGROUND/AIM: Cholangiocarcinoma is a devastating malignancy with limited treatment options and poor prognosis. Natural products have gained considerable attention for showing antitumor effects with less toxicities. Flavokawain B (FKB), a natural product, has been studied for its antitumor effects on various cancer cells. However, the anti-tumor effect of FKB on cholangiocarcinoma cells remains unknown. This study aimed at investigating the antitumor effect of FKB on cholangiocarcinoma cells in vitro and in vivo. MATERIALS AND METHODS: SNU-478, a human cholangiocarcinoma cell line, was used in this study. Effects of FKB on cell growth inhibition and apoptosis were investigated. The synergistic anti-tumor effect of FKB and cisplatin in combination was also evaluated. Western blotting was performed to examine the underlying molecular mechanisms of the effect of FKB. A xenograft mouse model study was performed to investigate the effect of FKB in vivo. RESULTS: FKB inhibited cell proliferation of cholangiocarcinoma cells in a concentration- and time-dependent manner. FKB also induced cellular apoptosis additively in combination with cisplatin. Akt pathway was suppressed by FKB either alone or in combination with cisplatin. In the xenograft model, FKB treatment in combination with cisplatin/gemcitabine significantly inhibited tumor growth of SNU-478 cells. CONCLUSION: FKB showed an antitumor effect through the induction of apoptosis, which was mediated by suppressing the Akt pathway in cholangiocarcinoma cells. However, the synergistic effect of FKB and cisplatin was not definite.

Causes for an extreme cold condition over Northeast Asia during April 2020.

Although 2020 was the fourth warmest year on record in northern Asia, the cold condition in April 2020 caused severe damage to the agricultural and marine ecosystems in northeastern Asia. Previous studies have indicated that the dipole atmospheric circulation over Siberia and the East Sea (Japan Sea) produced this cold environment with strong northwesterly wind. However, the potential causes of the dipole circulation over northeastern Asia remain unclear. In this study, we found that the East Atlantic/Western Russia (EAWR) pattern and blocking combined to produce the atmospheric structure. The wave train originated from the vorticity forcing of northwestern/central Russia and propagated from Western Europe to the East Sea via the background westerly and northerly winds. In addition, the Siberian blocking days increased eleven times in April 2020 relative to the climatological average, and an easterly (westerly) anomaly was observed over Mongolia-northeastern China (northern Russia). The strong blocking and EAWR pattern led to the robust atmospheric dipole structure with a prevailing northerly flow in April 2020, thereby causing the extreme cold condition over northeastern Asia. Our results provide novel insights into the cause of extreme cold condition in April over northeastern Asia.

The First Case of Infectious Spondylitis Caused by Gemella bergeri.

Gemella bergeri, a member of the genus Gemella, is a facultatively anaerobic, Gram-positive cocci. G. bergeri is a component of normal oral flora; however, it can become pathogenic and cause infections in patients with poor oral hygiene. A 78-year-old man was admitted to a hospital with a complaint of increasing posterior neck pain and lower back pain for 2 weeks. MRI was suggestive of infectious spondylitis at the C3-C4 level with prevertebral abscess formation, anterior epidural abscess formation. We identified Gemella bergeri in closed pus obtained during the surgery. Herein, we describe the first case of infective spondylitis caused by G. bergeri.

Stage-specific nutritional management and developmental programming to optimize meat production.

Over the past few decades, genetic selection and refined nutritional management have extensively been used to increase the growth rate and lean meat production of livestock. However, the rapid growth rates of modern breeds are often accompanied by a reduction in intramuscular fat deposition and increased occurrences of muscle abnormalities, impairing meat quality and processing functionality. Early stages of animal development set the long-term growth trajectory of offspring. However, due to the seasonal reproductive cycles of ruminant livestock, gestational nutrient deficiencies caused by seasonal variations, frequent droughts, and unfavorable geological locations negatively affect fetal development and their subsequent production efficiency and meat quality. Therefore, enrolling livestock in nutritional intervention strategies during gestation is effective for improving the body composition and meat quality of the offspring at harvest. These crucial early developmental stages include embryonic, fetal, and postnatal stages, which have stage-specific effects on subsequent offspring development, body composition, and meat quality. This review summarizes contemporary research in the embryonic, fetal, and neonatal development, and the impacts of maternal nutrition on the early development and programming effects on the long-term growth performance of livestock. Understanding the developmental and metabolic characteristics of skeletal muscle, adipose, and fibrotic tissues will facilitate the development of stage-specific nutritional management strategies to optimize production efficiency and meat quality.

AMP-activated protein kinase inhibition in fibro-adipogenic progenitors impairs muscle regeneration and increases fibrosis.

BACKGROUND: Following muscle injury, fibro-adipogenic progenitors (FAPs) are rapidly activated and undergo apoptosis at the resolution stage, which is required for proper muscle regeneration. When excessive FAPs remain, it contributes to fibrotic and fatty infiltration, impairing muscle recovery. Mechanisms controlling FAP apoptosis remain poorly defined. We hypothesized that AMP-activated protein kinase (AMPK) in FAPs mediates their apoptosis during the muscle regeneration. METHODS: To test, AMPKα1fl/fl PDGFRαCre mice were used to knock out AMPKα1 in FAPs. Following AMPKα1 knockout, the mice were injected with phosphate-buffered saline or glycerol to induce muscle injury. Tibialis anterior muscle and FAPs were collected at 3, 7 and 14 days post-injury (dpi) for further analysis. RESULTS: We found that AMPKα1 deletion in FAPs enhanced p65 translocation to the nuclei by 110% (n = 3; P < 0.01). AMPKα1 knockout group had a higher gene expression of MMP-9 (matrix metalloproteinase-9) by 470% (n = 3; P < 0.05) and protein level by 39% (n = 3; P < 0.05). Loss of AMPKα1 up-regulated the active TGF-ß1 (transforming growth factor-ß1) levels by 21% (n = 3; P < 0.05). TGF-ß promoted apoptotic resistance, because AMPKα1-deficient group had 36% lower cleaved Caspase 3 (cCAS3) content (n = 3; P < 0.05). Fibrotic differentiation of FAPs was promoted, with increased collagen protein level by 54% (n = 3; P < 0.05). Moreover, obesity decreased phosphorylation of AMPK by 54% (n = 3; P < 0.05), which decreased cCAS3 in FAPs by 44% (n = 3; P < 0.05) and elevated collagen accumulation (52%; n = 3; P < 0.05) during muscle regeneration. CONCLUSIONS: These data suggest that AMPK is a key mediator of FAPs apoptosis, and its inhibition due to obesity results in fibrosis of regenerated muscle.

Therapeutic carbohydrate restriction pre-COVID pandemic: assessing registered dietitians' knowledge, use and perceived barriers in Canada.

BACKGROUND/OBJECTIVES: Evidence supports therapeutic carbohydrate restriction (TCR) for managing appropriate patients with chronic illness, but little is known about TCR prescribing among dietitians. This study evaluated dietitians' knowledge, information use and needs for TCR in Canada. METHODS: Registered dietitians (RDs) were recruited (n = 274) from January to December 2020 to collect semi-structured data using an online needs assessment survey (French and English). Descriptive and inferential statistics were used to describe and assess which nine RD practice characteristics predicted TCR prescription in clinical practice. RESULTS: Respondents were located in all provinces and territories in Canada, with few international responses in the sample (3.5%). We found statistically significant differences between RDs who have prescribed TCR or not in four practice characteristics studied: level of knowledge (p < 0.001), reviewing literature (p = 0.02), clinician referrals (p < 0.001) or personal experience (p < 0.001). Multivariable models showed that the odds of prescribing TCR was associated with intermediate/expert knowledge (OR 5.92 [95% CI: 2.26-17.77]), clinician's referral (OR 3.22 [1.73-6.14]) and personal experience, whether a former user (OR 2.24 [1.09-4.72]) or a current user of TCR (OR 9.09 [2.70-42.09]), compared to no knowledge, no referral or no experience. CONCLUSION: There is a strong link between the use, or lack, of TCR in clinical practice among RDs and their knowledge level, personal experience and clinician referrals/support. Scope exists to develop novel educational tools and resources on scientific evidence for TCR, and increase multidisciplinary teams, so as to better support RDs in Canada to safely implement TCR in appropriate patients with chronic illness.