Accentuated Eccentric Loading and Alternative Set Structures: A Narrative Review for Potential Synergies in Resistance Training.

ABSTRACT: Chae, S, McDowell, KW, Baur, ML, Long, SA, Tufano, JJ, and Stone, MH. Accentuated eccentric loading and alternative set structures: A narrative review for potential synergies in resistance training. J Strength Cond Res 38(11): 1987-2000, 2024-As athletes become adapted to training over time, it becomes more difficult to develop their strength and power. In a conventional resistance training strategy, volume or load may be increased to provide novel stimuli to break through a plateau. However, physiological stress markers increase with increased volume or load, which is an innate shortcoming. In that case, practitioners strive to develop unconventional strategies that could increase training stimuli while adjusting fatigue. Two programming tactics, accentuated eccentric loading (AEL) using eccentric overload and alternative set structures (AS) using intraset rests, have been reported to increase training stimuli and alleviate fatigue, respectively. Importantly, when merging AEL and AS in various contexts, the 2 benefits could be accomplished together. Because AEL and AS cause different outcomes, it is important to deal with when and how they may be integrated into periodization. Moreover, prescribing eccentric overload and intraset rests requires logistical considerations that need to be addressed. This review discusses the scientific and practical aspects of AEL and AS to further optimize strength and power adaptations. This review discusses (a) scientific evidence as to which tactic is effective for a certain block, (b) potential practical applications, and (c) related discussions and future research directions.

Combined Accentuated Eccentric Loading and Rest Redistribution in High-Volume Back Squat: Acute Kinetics and Kinematics.

ABSTRACT: Chae, S, Long, SA, Lis, RP, McDowell, KW, Wagle, JP, Carroll, KM, Mizuguchi, S, and Stone, MH. Combined accentuated eccentric loading and rest redistribution in high-volume back squat: Acute kinetics and kinematics. J Strength Cond Res 38(4): 640-647, 2024-The purpose of this study was to explore acute kinetic and kinematic responses to combined accentuated eccentric loading and rest redistribution (AEL + RR). Resistance-trained men ( n = 12, 25.6 ± 4.4 years, 1.77 ± 0.06 m, and 81.7 ± 11.4 kg) completed a back squat (BS) 1 repetition maximum (1RM) and weight releaser familiarization session. Three BS exercise conditions (sets × repetitions × eccentric/concentric loading) consisted of (a) 3 × (5 × 2) × 110/60% (AEL + RR 5), (b) 3 × (2 × 5) × 110/60% (AEL + RR 2), and (c) 3 × 10 × 60/60% 1RM (traditional sets [TS]). Weight releasers (50% 1RM) were attached to every first repetition of each cluster set (every first, third, fifth, seventh, and ninth repetition in AEL + RR 5 and every first and sixth repetition in AEL + RR 2). The AEL + RR 5 resulted in significantly ( p < 0.05) greater concentric peak velocity (PV) (1.18 ± 0.17 m·s -1 ) and peak power (PP) (2,304 ± 499 W) compared with AEL + RR 2 (1.11 ± 0.19 m·s -1 and 2,148 ± 512 W) and TS (1.10 ± 0.14 m·s -1 and 2,079 ± 388 W). Furthermore, AEL + RR 5 resulted in significantly greater PV and PP across all 10 repetitions compared with TS. Although AEL + RR 5 resulted in significantly greater concentric mean force (MF) (1,706 ± 224 N) compared with AEL + RR 2 (1,697 ± 209 N) and TS (1,685 ± 211 N), no condition by set or repetition interactions existed. In conclusion, AEL + RR 5 increases PV and PP but has little effect on MF. Coaches might consider prescribing AEL + RR 5 to increase especially peak aspects of velocity and power outcomes.

Combined Accentuated Eccentric Loading and Rest Redistribution in High-Volume Back Squat: Acute Stimulus and Fatigue.

ABSTRACT: Chae, S, Long, SA, Lis, RP, McDowell, KW, Wagle, JP, Carroll, KM, Mizuguchi, S, and Stone, MH. Combined accentuated eccentric loading and rest redistribution in high-volume back squat: Acute stimulus and fatigue. J Strength Cond Res 38(4): 648-655, 2024-The purpose of this study was to examine acute stimulus and fatigue responses to combined accentuated eccentric loading and rest redistribution (AEL + RR). Resistance-trained men ( n = 12, 25.6 ± 4.4 years, 1.77 ± 0.06 m, and 81.7 ± 11.4 kg) completed a back squat (BS) 1 repetition maximum (1RM) and weight releaser familiarization session. Three BS exercise conditions (sets × repetitions × eccentric-concentric loading) consisted of (a) 3 × (5 × 2) × 110/60% (AEL + RR 5), (b) 3 × (2 × 5) × 110/60% (AEL + RR 2), and (c) 3 × 10 × 60/60% 1RM (traditional sets [TS]). Weight releasers (50% 1RM) were attached to every first repetition of each cluster set (every first, third, fifth, seventh, and ninth repetition in AEL + RR 5 and every first and sixth repetition in AEL + RR 2). The AEL + RR 5 resulted in greater total volume load (sets × repetitions × eccentric + concentric loading) (6,630 ± 1,210 kg) when compared with AEL + RR 2 (5,944 ± 1,085 kg) and TS (5,487 ± 1,002 kg). In addition, AEL + RR 5 led to significantly ( p < 0.05) greater rating of perceived exertion (RPE) after set 2 and set 3 and lower blood lactate (BL) after set 3 and 5, 15, and 25 minutes postexercise than AEL + RR 2 and TS. There was a main effect of condition for BL between AEL + RR 5 (5.11 ± 2.90 mmol·L -1 ), AEL + RR 2 (6.23 ± 3.22 mmol·L -1 ), and TS (6.15 ± 3.17 mmol·L -1 ). In summary, AEL + RR 5 results in unique stimulus and fatigue responses. Although it may increase perceived exertion, coaches could use AEL + RR 5 to achieve greater back squat total volume load while reducing BL accumulation.

Acute Physiological and Perceptual Responses to Rest Redistribution With Heavier Loads in Resistance-Trained Men.

ABSTRACT: Chae, S, Hill, DW, Bailey, CA, Moses, SA, McMullen, SM, and Vingren, JL. Acute physiological and perceptual responses to rest redistribution with heavier loads in resistance-trained men. J Strength Cond Res 37(5): 994-1000, 2023-The purpose of this study was to explore the effect of rest redistribution with heavier loads (RR + L) on physiological and perceptual responses in resistance-trained men. Eight men who had back squat (BS) 1 repetition maximum (1RM) to body mass ratio; 1.8 ± 0.2 completed 2 BS exercise sessions in a counterbalanced and a randomized order; RR + L: 4 sets of (2 × 5) repetitions with 90-second interset rest and 30-second intraset rest using 75% BS 1RM and traditional sets (TS): 4 sets of 10 repetitions with 120-second interset rest using 70% BS 1RM. Blood samples were collected before exercise, immediately post exercise, and 5, 15, and 30 minutes post exercise for the analysis of growth hormone (GH), total testosterone (TT), cortisol (C), and blood lactate (BL), whereas rating of perceived exertion (RPE) and heart rate (HR) were measured immediately after each set of the BS exercise. While neither main effect of condition nor interaction existed, there was a significant ( p < 0.05) main effect of time point (and set) for GH, TT, C, BL, RPE, and HR. Volume load was greater for RR + L compared with TS (4,074.9 ± 786.7 kg vs. 3,796.3 ± 714.8 kg). In conclusion, RR + L increases volume load by approximately 7% but does not seem to influence GH ( g = -0.15), TT ( g = -0.09), BL ( g = -0.22), RPE ( g = 0.14), and HR ( g = -0.08) responses. Practitioners may consider using RR + L to increase volume load without increasing acute fatigue responses.

Complications of Nasopharyngeal Swabs and Safe Procedures for COVID-19 Testing Based on Anatomical Knowledge.

Nasopharyngeal swabs have been widely to prevent the spread of coronavirus disease 2019 (COVID-19). Nasopharyngeal COVID-19 testing is a generally safe and well-tolerated procedure, but numerous complications have been reported in the media. Therefore, the present study aimed to review and document adverse events and suggest procedural references to minimize preventable but often underestimated risks. A total of 27 articles were selected for the review of 842 related documents in PubMed, Embase, and KoreaMed. The complications related to nasopharyngeal COVID-19 testing were reported to be rarely happened, ranging from 0.0012 to 0.026%. Frequently documented adverse events were retained swabs, epistaxis, and cerebrospinal fluid leakage, often associated with high-risk factors, including severe septal deviations, pre-existing skull base defects, and previous sinus or transsphenoidal pituitary surgery. Appropriate techniques based on sufficient anatomical knowledge are mandatory for clinicians to perform nasopharyngeal COVID-19 testing. The nasal floor can be predicted by the line between the nostril and external ear canal. For safe testing, the angle of swab insertion in the nasal passage should remain within 30° of the nasal floor. The swab was gently inserted along the nasal septum just above the nasal floor to the nasopharynx and remained on the nasopharynx for several seconds before removal. Forceful insertion should be attempted, and alternative examinations should be considered, especially in vulnerable patients. In conclusion, patients and clinicians should be aware of rare but possible complications and associated high-risk factors. The suggested procedural pearls enable more comfortable and safe nasopharyngeal COVID-19 testing for both clinicians and patients.

Osteogenic Efficacy of Human Trophoblasts-Derived Conditioned Medium on Mesenchymal Stem Cells.

Trophoblasts play an important role in the regulation of the development and function of the placenta. Our recent study demonstrated the skin regeneration capacity of trophoblast-derived extracellular vesicles (EV). Here, we aimed to determine the potential of trophoblast-derived conditioned medium (TB-CM) in enhancing the osteogenic differentiation of bone marrow mesenchymal stem cells (MSCs). We found that TB-CM promoted the osteogenic differentiation of MSCs in a dose-dependent manner. Furthermore, it inhibited adipogenesis of MSCs. We also found that the primary trophoblast-derived conditioned medium (PTB-CM) significantly enhanced the proliferation and osteogenic differentiation of human MSCs. Our study demonstrated the regulatory mechanisms underlying the TB-CM-induced osteogenesis in MSCs. An upregulation of genes associated with cytokines/chemokines was observed. The treatment of MSCs with TB-CM stimulated osteogenesis by activating several biological processes, such as mitogen-activated protein kinase (MAPK) and bone morphogenetic protein 2 (BMP2) signaling. This study demonstrated the proliferative and osteogenic efficacies of the trophoblast-derived secretomes, suggesting their potential for use in clinical interventions for bone regeneration and treatment.

Extracellular Vesicles (Secretomes) from Human Trophoblasts Promote the Regeneration of Skin Fibroblasts.

To date, placental trophoblasts have been of interest in the fields of obstetrics and gynecology, mainly due to their involvement in the formation of a connection between the mother and fetus that aids in placental development and fetal survival. However, the regenerative capacities of trophoblasts for application in regenerative medicine and tissue engineering are poorly understood. Here, we aim to determine the skin regeneration and anti-aging capacities of trophoblast-derived conditioned medium (TB-CM) and exosomes (TB-Exos) using human normal dermal fibroblasts (HNDFs). TB-CM and TB-Exos treatments significantly elevated the migration and proliferation potencies of HNDF cells in a dose- and time-dependent manner. When RNA sequencing (RNA-seq) was used to investigate the mechanism underlying TB-CM-induced cell migration on scratch-wounded HNDFs, the increased expression of genes associated with C-X-C motif ligand (CXCL) chemokines, toll-like receptors, and nuclear factor-kappa B (NF-κB) signaling was observed. Furthermore, treatment of intrinsically/extrinsically senescent HNDFs with TB-CM resulted in an enhanced rejuvenation of HNDFs via both protection and restoration processes. Gene expression of extracellular matrix components in the skin dermis significantly increased in TB-CM- and TB-Exos-treated HNDFs. These components are involved in the TB-CM and Exo-mediated regeneration and anti-aging of HNDFs. Thus, this study demonstrated the regenerative and anti-aging efficacies of trophoblast-derived secretomes, suggesting their potential for use in interventions for skin protection and treatment.

Prevalence of Dizziness and Associated Factors in South Korea: A Cross-Sectional Survey From 2010 to 2012.

BACKGROUND: Dizziness is one of the most common complaints in medicine and a frequent symptom among older people. Dizziness has a considerable impact on life quality and is associated with high economic costs. The purpose of this study was to evaluate the prevalence of dizziness in the general population and to describe its clinical characteristics and associated factors. METHODS: The Korea National Health and Nutrition Examination Survey (KNHANES) is a cross-sectional survey of the civilian, non-institutionalized population of South Korea. We evaluated data for 12,653 participants (5,450 men and 7,203 women), aged 40 years and above, whom participated in the KNHANES between 2010 and 2012. RESULTS: In the age group over 40 years old, the 1-year prevalence of dizziness was 20.10%. Dizziness was more prevalent among women (25.18%) than among men (14.57%; P < 0.001) and the prevalence rate increased with age (P < 0.001). In multivariable analysis, female sex, older age, serum triglyceride level, experience of depression, limited functional status owing to visual acuity impairment, limited physical performance, smoking, alcohol consumption, and perception of stress were independently associated with dizziness. CONCLUSIONS: In our study, the prevalence of dizziness in the general population was 20.10%. There was a stronger relationship between dizziness and physical performance, chronic diseases, and health behaviors compared to that with otologic diseases. Interventions for dizziness should be approached in a multifactorial manner and an understanding of various factors is necessary for the prevention and management of this condition.

A Trend in Sudden Sensorineural Hearing Loss: Data from a Population-Based Study.

This is the first study reporting on the incidence and clinical aspects of sudden sensorineural hearing loss (SSNHL) in South Korea. Using Health Insurance Review and Assessment Service data from 2011 to 2015, the monthly incidence of unilateral SSNHL and incidence according to patients' sex, age, and month of diagnosis were investigated. The monthly incidence of unilateral SSNHL increased over the 5-year study period, with a mean annual incidence of 17.76 cases/ 100,000 of the population. The incidence increased with age, with most patients presenting in their 60s. There was a slight female preponderance, with a male-to-female ratio of 1: 1.35. Most new patients were diagnosed in October, and the fewest in January. In conclusion, this large-scale study indicates that unilateral SSNHL has a higher incidence among the elderly, women, and in autumn (i.e., along with colder weather).

Mucosa-associated Lymphoid Tissue Misdiagnosis as Glomus Tympanicum.

Middle ear tumors are diverse, but relatively uncommon. The most frequent tumor in the middle ear is glomus tumor, followed by others such as schwannoma and cholesteatoma. We experienced a case of Mucosa-associated lymphoid tissue hyperplasia as a middle ear tumor. The mass behind tympanic membrane appeared a hypervascular tumor, mimicking a glomus tumor, but the form of multiple separate masses in middle ear and mastoid cavity was the distinguishing feature that set it apart from a glomus tumor. Additionally, another characteristic was its tendency to easily shrink under pressure. This characteristic should be considered when encounter a hypervascular looking middle ear mass. Laryngoscope, 134:1894-1896, 2024.

Changes in Otitis Media During COVID-19.

OBJECTIVES: During the COVID-19 pandemic, various non-pharmaceutical interventions such as individual hygiene practices like hand washing, social distancing, and mandates for the use of masks in public spaces were implemented to reduce the spread of the disease. Otitis media (OM) is a common infectious disease. How the changed environment due to the COVID-19 pandemic has influenced the prevalence of infectious diseases like OM is not known. This study aimed to investigate how OM prevalence and trends changed during COVID-19 in Korea. METHODS: OM patient data from 2017 to 2021 were extracted from the Health Insurance Review and Assessment Service database. Patients diagnosed with disease code H66 (suppurative and unspecified otitis media) were selected for analysis. Data on OM prevalence, gender, region, medical institution, and number of ventilating-tube prescriptions were analyzed. All age groups were included, and ages were categorized into 5-year ranges. RESULTS: The number of patients diagnosed with the OM disease code decreased continuously from 2017 to 2021 (1 598 205, 1 560 178, 1 520 948, 983 701, and 734 901). The average OM prevalence per 1000 persons decreased by 45.0% from 30.2 in 2017 to 2019 to 16.6 in 2020 to 2021. The change of OM prevalence was greater for the 0 to 5 age group than other age groups. The decrease in average prevalence per 1000 persons was greatest in the 0 to 5 age group (48.6% decrease from 358.2 in 2017-2019 to 184.1 in 2020-2021). The impact of environmental changes on ventilation-tube insertion was smaller than on OM prevalence. The average number of ventilating-tube insertions decreased by 28.1% from 27 311 in 2017 to 2019 to 19 650 in 2020 to 2021. CONCLUSIONS: OM prevalence decreased by 45.0%, and the number of ventilating-tube insertions decreased by 28.1% in Korea during COVID-19.

Association of the protective effect of telmisartan on hearing loss among patients with hypertension.

Aim: Hearing loss, affecting a significant portion of the global population, is prevented with peroxisome proliferator-activated receptor γ agonism. Understanding potential protective treatments is crucial for public health. We examine the effect of telmisartan, an antihypertensive drug and partial peroxisome proliferator-activated receptor γ agonist, on hearing loss in patients with hypertension. Method and results: This retrospective cohort analysis used data from the OMOP Common Data Model database, encompassing information from three tertiary institutions in South Korea. The study included a substantial sample size of 860,103 people diagnosed with hypertension. The study included individuals who had been medically diagnosed with hypertension and had been prescribed antihypertensive drugs, including telmisartan. The study design was established to evaluate the comparative effects of telmisartan and other hypertension medications on hearing loss. We used propensity score matching (PSM) to create a balanced cohort, reducing potential biases between the telmisartan and non-telmisartan groups. From the initial 860,103 patients with hypertension, a propensity score matched cohort was derived from 20,010 patients, with 2,193 in the telmisartan group. After PSM, lower incidence of total hearing loss was observed in the telmisartan group compared to the non-telmisartan group during the 3-year follow-up (0.5% vs. 1.5%, log-rank p = 0.005). In subgroup analysis, this study showed consistent results that lower incidence of total hearing loss was higher in the telmisartan group than in the non-telmisartan group. Conclusion: Telmisartan was associated with reducing certain types of hearing loss in patients with hypertension. Further research is needed to confirm these findings and understand the mechanisms.

Caffeic acid phenethyl ester inhibits diesel exhaust particle-induced inflammation of human middle ear epithelial cells via NOX4 inhibition.

OBJECTIVES: Otitis media is one of the most common diseases in pediatric populations. Recent research on its pathogenesis has focused on air pollution. Chronic exposure to particulate air pollution is associated with the impairment of middle ear function. However, the mechanisms and the underlying inhibitory pathways, especially in the human middle ear, remain unknown. Caffeic acid phenethyl ester (CAPE) is a biologically active ingredient of propolis, a product of honeybee hives, which has anti-oxidative and anti-inflammatory activities. The aim of this study was to evaluate the inhibitory effect of CAPE on diesel exhaust particle (DEP)-induced inflammation of human middle ear epithelial cells and to determine the underlying pathway of the action of CAPE. METHODS: The inflammatory damage caused by DEPs and the anti-inflammatory effects of CAPE were determined by measuring the levels of tumor necrosis factor alpha and nicotinamide adenine dinucleotide phosphate oxidase (NOX) 4 with real-time reverse transcription polymerase chain reaction and Western blot analysis. The oxidative stress induced by DEPs and the anti-oxidative effects of CAPE were directly evaluated by measuring reactive oxygen species production by use of flow cytometric analysis of 2',7'-dichlorofluorescein diacetate. The effects of CAPE were compared with those of N-acetyl-L-cysteine, which has anti-oxidative and anti-inflammatory effects. RESULTS: Use of CAPE significantly inhibited DEP-induced up-regulation of tumor necrosis factor alpha and NOX4 expression in a dose- and time-dependent manner. The accumulation of reactive oxygen species induced by DEPs was decreased by pretreatment with CAPE. The anti-inflammatory and anti-oxidative effects of CAPE were similar to those of N-acetyl-L-cysteine. CONCLUSIONS: The inflammation induced by DEP is reduced by CAPE via the inhibition of NOX4 expression. These findings suggest that CAPE might be used as a therapeutic agent against DEP-induced inflammation of human middle ear epithelial cells.

Regenerative capacity of trophoblast stem cell-derived extracellular vesicles on mesenchymal stem cells.

BACKGROUND: Human mesenchymal stem cells (MSCs) are therapeutic for clinical applications because of their excellent immunomodulatory and multiple lineage differentiation abilities at tissue injury sites. However, insufficient number of cells and lack of regenerative properties during in vitro expansion still limit the clinical applicability of MSC therapies. Here, we demonstrated a preconditioning strategy with trophoblast stem cell-derived extracellular vesicles (TSC-EVs) to boost the proliferation and regenerative capacity of MSCs. METHODS: We employed cell proliferation analyses such as CCK8 and BrdU assays to determine the proliferation-promoting role of TSC-EVs on MSCs. Osteogenic effects of TSC-EVs on MSCs were assessed by alkaline phosphatase (ALP) activity, calcium assays, and calvarial bone defect animal models. For skin regenerative effects, skin wound mice model was exploited to analyze wound-healing rate in this study, as well as immunofluorescence and histological staining evaluates. We also performed the small RNA profiling and RNA-sequencing analyzes to understand the cellular mechanism of TSC-EVs on MSCs. RESULTS: TSC-EVs significantly promoted MSC proliferation under xeno-free conditions and facilitated the therapeutic effects of MSCs, including osteogenesis, anti-senescence, and wound healing. Transcriptomic analysis also provided evidence that specific microRNAs in TSC-EVs and differentially expressed genes (DEGs) in TSC-EV-treated MSCs showed the possibility of TSC-EVs triggering the regenerative abilities of MSCs with cytokine interaction. Hence, we found that NGF/Akt signaling mediated the regenerative effects of TSC-EVs on MSCs as a particular cellular signaling pathway. CONCLUSION: The results of this study demonstrated the functional properties of TSC-EVs on MSCs for MSC-based therapeutic applications, suggesting that TSC-EVs may serve as a potential preconditioning source for MSC therapy in the clinical field of regenerative medicine.

Hydrophilic and lipophilic statin use and risk of hearing loss in hyperlipidemia using a Common Data Model: multicenter cohort study.

Hearing impairment, the third largest health burden worldwide, currently lacks definitive treatments or preventive drugs. This study compared the effects of hydrophilic and lipophilic statin on hearing loss using a common database model. This retrospective multicenter study was conducted in three hospitals in South Korea (Anam, Guro, Ansan). We enrolled patients with hyperlipidemia with an initial hearing loss diagnosis. Data were collected during January 1, 2022-December 31, 2021 using the Observational Health Data Science and Informatics open-source software and Common Data Model database. The primary outcome was the occurrence of first-time hearing loss following a hyperlipidemia diagnosis, as documented in the Common Data Model cohort database. The measures of interest were hearing loss risk between hydrophilic and lipophilic statin use. Variables were compared using propensity score matching, Cox proportional regression, and meta-analysis. Among 37,322 patients with hyperlipidemia, 13,751 (7669 men and 6082 women) and 23,631 (11,390 men and 12,241 women) were treated with hydrophilic and lipophilic statins, respectively. After propensity score matching, according to the Kaplan-Meier curve, hearing loss risk did not significantly differ among the hospitals. The hazard ratio (HR) of the male patients from Anam (0.29, [95% confidence interval (CI), 0.05-1.51]), Guro (HR, 0.56, [95% CI 0.18-1.71]), and Ansan (hazard ratio, 0.29, [95% CI 0.05-1.51]) hospitals were analyzed using Cox proportional regression. Overall effect size (HR, 0.40, [95% CI 0.18-0.91]) was estimated using meta-analysis, which indicated that hearing loss risk among hydrophilic statin users was less than that among lipophilic statin users and was statistically significant. Men in the hydrophilic statin group had a lower risk of hearing impairment than those in the lipophilic statin group.

Effect of 5-azacytidine on in vitro biofilm formation of Streptococcus pneumoniae.

Streptococcus pneumoniae is a gram-positive bacterium that causes otitis media, pneumonia, meningitis and sepsis in young children and the elderly. Previous studies reported that pneumococci in different diseases do not behave as planktonic cells, but predominantly show characteristics of a biofilm. In this study we examine the effect of 5-azacytidine on S. pneumoniae, particularly on biofilm formation and investigate the gene expression involved in synthesis of autoinducer-2, competence and DNA repair. The effect of 5-aza on in vitro biofilm formation was studied by the crystal violet microtiter plate method. The S. pneumoniae biofilms were grown with different concentration of 5-azacytidine (15-500 µm), at variable time intervals and the inhibition percentages were calculated. The effects of 5-aza on the morphology of biofilms were analyzed by scanning electron microscope. The relative quantification of 11 genes of biofilms grown with 5-aza involved in autoinducer-2 synthesis, competence and DNA repair was carried out by real-time RT-PCR with respect to biofilms grown without 5-aza. The crystal violet microtiter assay detected a significant inhibitory effect of 5-aza on in vitro biofilm formation, at concentration that did not inhibited planktonic cell growth. The SEM analysis demonstrated thin and disrupted biofilms, without micro-colonies in the samples treated with 5-aza, while these structures were present in the biofilms grown without 5-aza. The relative quantification of gene expression of 5-aza biofilms showed a significant down regulation of genes involved in the methionine and homocysteine recycling pathway which produces quorum sensing molecule autoinducer-2 as by-products. A significant decrease in the expressions of luxS, metK, pfs and cmK was detected. In conclusion, 5-aza inhibits in vitro biofilm formation and decreases the expression of luxS, pfs and metK, which are involved in the synthesis of autoinducer-2 as by-products of the methionine recycling pathway. The inhibitory effect of 5-aza may be either due to down regulation of pfs, luxS and metK or due to accumulation of the toxic substrate of pfs, luxS and metK genes.

Gene expression profile of early in vitro biofilms of Streptococcus pneumoniae.

In this study, the gene expression profile of early in vitro Streptococcus pneumoniae biofilm with respect to planktonic cells in cDNA microarray analysis is reported. Microarray analysis with respect to planktonic cells was performed on total RNA extracted from biofilms grown in 24-well microtiter plates. To validate the microarray results, real-time RT-PCR was performed on 13 differentially expressed genes and one constitutively expressed gene. The cDNA-microarray analyses identified 89 genes that were significantly differentially expressed in biofilm and planktonic cells. Genes involved in isoprenoid biosynthesis, cell wall biosynthesis, translation and purine and pyrimidine nucleotide metabolic pathways were exclusively expressed in the biofilms, whereas transcription regulator genes were exclusively expressed in planktonic cells. The real-time RT-PCR results of 13 differentially regulated genes were completely in agreement with the microarray data. The exclusive up regulation in biofilms of genes involved in the mevalonate pathway, cell wall biosynthesis, translation and purine and pyrimidine nucleotide metabolic pathways suggests that expression of these genes may be required for initial biofilm formation, and growth and survival of bacteria in biofilms. The up regulation of related genes suggests that cells in biofilms may be under stress conditions and possibly actively involved in the protein synthesis required to adapt to a new environment.

Comprehensive analysis of human chorionic membrane extracts regulating mesenchymal stem cells during osteogenesis.

OBJECTIVE: Human chorionic membrane extracts (CMEs) from placenta are known to be a natural biomaterial for bone regeneration, with their excellent osteogenic efficacy on osteoblasts. However, little is known about the regulatory mechanism involved. METHODS AND RESULTS: We have shown the in vitro and in vivo bone-forming ability of CME using human osteoblasts and bone defect animal models, suggesting that CME greatly enhances osteogenesis by providing an osteoconductive environment for the osteogenesis of osteoblasts. Proteomic analysis revealed that CME contained several osteogenesis-related stimulators such as osteopontin, osteomodulin, Thy-1, netrin 4, retinol-binding protein and DJ-1. Additionally, 23 growth factors/growth factor-related proteins were found in CME, which may trigger mitogen-activated protein kinase (MAPK) signalling as a specific cellular signalling pathway for osteogenic differentiation. Microarray analysis showed four interaction networks (chemokine, Wnt signalling, angiogenesis and ossification), indicating the possibility that CME can promote osteogenic differentiation through a non-canonical Wnt-mediated CXCL signalling-dependent pathway. CONCLUSIONS: The results of this study showed the function and mechanism of action of CME during the osteogenesis of osteoblasts and highlighted a novel strategy for the use of CME as a biocompatible therapeutic material for bone regeneration.

Different Transcutaneous Auricular Vagus Nerve Stimulation Parameters Modulate the Anti-Inflammatory Effects on Lipopolysaccharide-Induced Acute Inflammation in Mice.

Vagus nerve stimulation (VNS) is considered a potential method for anti-inflammation due to the involvement of the VN in the cholinergic anti-inflammatory pathway (CAP) formation of a connection between the central nervous system and peripheral immune cells that help relieve inflammation. However, whether a non-invasive transcutaneous auricular VNS (taVNS) modulates the inflammation levels via altering the parameter of taVNS is poorly understood. This study aimed to determine the differential inhibitory effects of taVNS on lipopolysaccharide (LPS)-induced systemic inflammation using electrical stimulation parameters such as pulse frequency and time. The taVNS-promoted CAP activity significantly recovered LPS-induced tissue injuries (lung, spleen, and intestine) and decreased inflammatory cytokine levels and tissue-infiltrated immune cells. Interestingly, the anti-inflammatory capacity of taVNS with 15 Hz was much higher than that of taVNS with 25 Hz. When a cytokine array was used to investigate the changes of inflammation and immune response-related cytokines/chemokines expression in taVNS with 15 Hz or 25 Hz treatment in LPS-induced endotoxemia in mice, most of the expression of cytokines/chemokines associated with pro-inflammation was severely decreased in taVNS with 15 Hz compared to 25 Hz. This study demonstrated that the taVNS parameter could differentially modulate the inflammation levels of animals, suggesting the importance of taVNS parameter selection for use in feasible interventions for acute inflammation treatment.