Gut microbiome predicts cognitive function and depressive symptoms in late life.

Depression in older adults with cognitive impairment increases progression to dementia. Microbiota is associated with current mood and cognition, but the extent to which it predicts future symptoms is unknown. In this work, we identified microbial features that reflect current and predict future cognitive and depressive symptoms. Clinical assessments and stool samples were collected from 268 participants with varying cognitive and depressive symptoms. Seventy participants underwent 2-year follow-up. Microbial community diversity, structure, and composition were assessed using high-resolution 16 S rRNA marker gene sequencing. We implemented linear regression to characterize the relationship between microbiome composition, current cognitive impairment, and depressive symptoms. We leveraged elastic net regression to discover features that reflect current or future cognitive function and depressive symptoms. Greater microbial community diversity associated with lower current cognition in the whole sample, and greater depression in participants not on antidepressants. Poor current cognitive function associated with lower relative abundance of Bifidobacterium, while greater GABA degradation associated with greater current depression severity. Future cognitive decline associated with lower cognitive function, lower relative abundance of Intestinibacter, lower glutamate degradation, and higher baseline histamine synthesis. Future increase in depressive symptoms associated with higher baseline depression and anxiety, lower cognitive function, diabetes, lower relative abundance of Bacteroidota, and lower glutamate degradation. Our results suggest cognitive dysfunction and depression are unique states with an overall biological effect detectable through gut microbiota. The microbiome may present a noninvasive readout and prognostic tool for cognitive and psychiatric states.

Pregnancy loss in major fetal congenital heart disease: incidence, risk factors and timing.

OBJECTIVE: Fetuses with congenital heart disease (CHD) are at increased risk of pregnancy loss compared with the general population. We aimed to assess the incidence, timing and risk factors of pregnancy loss in cases with major fetal CHD, overall and according to cardiac diagnosis. METHODS: This was a retrospective, population-level cohort study of fetuses and infants diagnosed with major CHD between 1997 and 2018 identified by the Utah Birth Defect Network (UBDN), excluding cases with termination of pregnancy and minor cardiovascular diagnoses (e.g. isolated aortic/pulmonary pathology and isolated septal defects). The incidence and timing of pregnancy loss were recorded, overall and according to CHD diagnosis, with further stratification based on presence of isolated CHD vs additional fetal diagnosis (genetic diagnosis and/or extracardiac malformation). Adjusted risk of pregnancy loss was calculated and risk factors were assessed using multivariable models for the overall cohort and prenatal diagnosis subgroup. RESULTS: Of 9351 UBDN cases with a cardiovascular code, 3251 cases with major CHD were identified, resulting in a study cohort of 3120 following exclusion of cases with pregnancy termination (n = 131). There were 2956 (94.7%) live births and 164 (5.3%) cases of pregnancy loss, which occurred at a median gestational age of 27.3 weeks. Of study cases, 1848 (59.2%) had isolated CHD and 1272 (40.8%) had an additional fetal diagnosis, including 736 (57.9%) with a genetic diagnosis and 536 (42.1%) with an extracardiac malformation. The observed incidence of pregnancy loss was highest in the presence of mitral stenosis (< 13.5%), hypoplastic left heart syndrome (HLHS) (10.7%), double-outlet right ventricle with normally related great vessels or not otherwise specified (10.5%) and Ebstein's anomaly (9.9%). The adjusted risk of pregnancy loss was 5.3% (95% CI, 3.7-7.6%) in the overall CHD population and 1.4% (95% CI, 0.9-2.3%) in cases with isolated CHD (adjusted risk ratio, 9.0 (95% CI, 6.0-13.0) and 2.0 (95% CI, 1.0-6.0), respectively, based on the general population risk of 0.6%). On multivariable analysis, variables associated with pregnancy loss in the overall CHD population included female fetal sex (adjusted odds ratio (aOR), 1.6 (95% CI, 1.1-2.3)), Hispanic ethnicity (aOR, 1.6 (95% CI, 1.0-2.5)), hydrops (aOR, 6.7 (95% CI, 4.3-10.5)) and additional fetal diagnosis (aOR, 6.3 (95% CI, 4.1-10)). On multivariable analysis of the prenatal diagnosis subgroup, years of maternal education (aOR, 1.2 (95% CI, 1.0-1.4)), presence of an additional fetal diagnosis (aOR, 2.7 (95% CI, 1.4-5.6)), atrioventricular valve regurgitation ≥ moderate (aOR, 3.6 (95% CI, 1.3-8.8)) and ventricular dysfunction (aOR, 3.8 (95% CI, 1.2-11.1)) were associated with pregnancy loss. Diagnostic groups associated with pregnancy loss were HLHS and variants (aOR, 3.0 (95% CI, 1.7-5.3)), other single ventricles (aOR, 2.4 (95% CI, 1.1-4.9)) and other (aOR, 0.1 (95% CI, 0-0.97)). Time-to-pregnancy-loss analysis demonstrated a steeper survival curve for cases with an additional fetal diagnosis, indicating a higher rate of pregnancy loss compared to cases with isolated CHD (P < 0.0001). CONCLUSIONS: The risk of pregnancy loss is higher in cases with major fetal CHD compared with the general population and varies according to CHD type and presence of additional fetal diagnoses. Improved understanding of the incidence, risk factors and timing of pregnancy loss in CHD cases should inform patient counseling, antenatal surveillance and delivery planning. © 2023 International Society of Ultrasound in Obstetrics and Gynecology.

Femtosecond response of polyatomic molecules to ultra-intense hard X-rays.

X-ray free-electron lasers enable the investigation of the structure and dynamics of diverse systems, including atoms, molecules, nanocrystals and single bioparticles, under extreme conditions. Many imaging applications that target biological systems and complex materials use hard X-ray pulses with extremely high peak intensities (exceeding 1020 watts per square centimetre). However, fundamental investigations have focused mainly on the individual response of atoms and small molecules using soft X-rays with much lower intensities. Studies with intense X-ray pulses have shown that irradiated atoms reach a very high degree of ionization, owing to multiphoton absorption, which in a heteronuclear molecular system occurs predominantly locally on a heavy atom (provided that the absorption cross-section of the heavy atom is considerably larger than those of its neighbours) and is followed by efficient redistribution of the induced charge. In serial femtosecond crystallography of biological objects-an application of X-ray free-electron lasers that greatly enhances our ability to determine protein structure-the ionization of heavy atoms increases the local radiation damage that is seen in the diffraction patterns of these objects and has been suggested as a way of phasing the diffraction data. On the basis of experiments using either soft or less-intense hard X-rays, it is thought that the induced charge and associated radiation damage of atoms in polyatomic molecules can be inferred from the charge that is induced in an isolated atom under otherwise comparable irradiation conditions. Here we show that the femtosecond response of small polyatomic molecules that contain one heavy atom to ultra-intense (with intensities approaching 1020 watts per square centimetre), hard (with photon energies of 8.3 kiloelectronvolts) X-ray pulses is qualitatively different: our experimental and modelling results establish that, under these conditions, the ionization of a molecule is considerably enhanced compared to that of an individual heavy atom with the same absorption cross-section. This enhancement is driven by ultrafast charge transfer within the molecule, which refills the core holes that are created in the heavy atom, providing further targets for inner-shell ionization and resulting in the emission of more than 50 electrons during the X-ray pulse. Our results demonstrate that efficient modelling of X-ray-driven processes in complex systems at ultrahigh intensities is feasible.

Effect of varied dorsiflexion range of motion on landing biomechanics in chronic ankle instability.

BACKGROUND: Limited dorsiflexion range of motion (DFROM) is a risk factor for lateral ankle sprain. However, varied DFROM exists within the chronic ankle instability (CAI) population, and how the variability may influence altered movement patterns during landing is unclear. OBJECTIVE: The purpose of this study was to identify different movement strategies during maximal jump landing/cutting among CAI patients classified by varied DFROM. METHODS: One hundred CAI subjects were classified into 3 subgroups based on their DFROM, measured by the weight-bearing lunge test: a Hypo- (≤40°), Normal- (40-50°), and Hyper-DFROM group (≥50°). Participants completed five trials of maximal jump landing/cutting. Lower extremity joint angles and EMG activation of seven muscles were collected from initial contact to toe-off. Functional analyses of variance were used to evaluate between-group differences for these outcome variables. RESULTS: Hypo-DFROM group (14M, 10F) displayed the reduced ankle dorsiflexion and inversion angles with increased hip flexion angle as a compensatory kinematic chain movement strategy. In addition, motion restrictions of the ankle are associated with altered muscle activation in both distal and proximal muscles during landing/cutting. Normal-DFROM (25M, 30F) and Hyper-DFROM (11M, 10F) groups also have different movement strategies including greater inversion angle and less EMG activation, which could contribute to further ankle injuries. CONCLUSIONS: Our data suggest that limited DFROM negatively affects the ankle joint during demanding movement within the CAI population. These movement patterns in CAI patients with pathomechanical deficits could contribute to further ankle sprains.

A Review of the Relationships Between Knee Pain and Movement Neuromechanics.

CONTEXT: Knee injury and disease are common, debilitating, and expensive. Pain is a chief symptom of knee injury and disease and likely contributes to arthrogenic muscle inhibition. Joint pain alters isolated motor function, muscular strength, and movement biomechanics. Because knee pain influences biomechanics, it likely also influences long-term knee joint health. OBJECTIVE: The purpose of this article is 2-fold: (1) review effects of knee pain on lower-extremity muscular activation and corresponding biomechanics and (2) consider potential implications of neuromechanical alterations associated with knee pain for long-term knee joint health. Experimental knee pain is emphasized because it has been used to mimic clinical knee pain and clarify independent effects of knee pain. Three common sources of clinical knee pain are also discussed: patellofemoral pain, anterior cruciate ligament injury and reconstruction, and knee osteoarthritis. DATA SOURCES: The PubMed, Web of Science, and SPORTDiscus databases were searched for articles relating to the purpose of this article. CONCLUSION: Researchers have consistently reported that knee pain alters neuromuscular activation, often in the form of inhibition that likely occurs via voluntary and involuntary neural pathways. The effects of knee pain on quadriceps activation have been studied extensively. Knee pain decreases voluntary and involuntary quadriceps activation and strength and alters the biomechanics of various movement tasks. If allowed to persist, these neuromechanical alterations might change the response of articular cartilage to joint loads during movement and detrimentally affect long-term knee joint health. Physical rehabilitation professionals should consider neuromechanical effects of knee pain when treating knee injury and disease. Resolution of joint pain can likely help to restore normal movement neuromechanics and potentially improve long-term knee joint health and should be a top priority.

Pulse Energy and Pulse Duration Effects in the Ionization and Fragmentation of Iodomethane by Ultraintense Hard X Rays.

The interaction of intense femtosecond x-ray pulses with molecules sensitively depends on the interplay between multiple photoabsorptions, Auger decay, charge rearrangement, and nuclear motion. Here, we report on a combined experimental and theoretical study of the ionization and fragmentation of iodomethane (CH_{3}I) by ultraintense (∼10^{19} W/cm^{2}) x-ray pulses at 8.3 keV, demonstrating how these dynamics depend on the x-ray pulse energy and duration. We show that the timing of multiple ionization steps leading to a particular reaction product and, thus, the product's final kinetic energy, is determined by the pulse duration rather than the pulse energy or intensity. While the overall degree of ionization is mainly defined by the pulse energy, our measurement reveals that the yield of the fragments with the highest charge states is enhanced for short pulse durations, in contrast to earlier observations for atoms and small molecules in the soft x-ray domain. We attribute this effect to a decreased charge transfer efficiency at larger internuclear separations, which are reached during longer pulses.

Stillbirth and fetal anomalies: secondary analysis of a case-control study.

OBJECTIVE: Approximately 10% of stillbirths are attributed to fetal anomalies, but anomalies are also common in live births. We aimed to assess the relationship between anomalies, by system and stillbirth. DESIGN: Secondary analysis of a prospective, case-control study. SETTING: Multicentre, 59 hospitals in five regional catchment areas in the USA. POPULATION OR SAMPLE: All stillbirths and representative live birth controls. METHODS: Standardised postmortem examinations performed in stillbirths, medical record abstraction for stillbirths and live births. MAIN OUTCOME MEASURES: Incidence of major anomalies, by type, compared between stillbirths and live births with univariable and multivariable analyses using weighted analysis to account for study design and differential consent. RESULTS: Of 465 singleton stillbirths included, 23.4% had one or more major anomalies compared with 4.3% of 1871 live births. Having an anomaly increased the odds of stillbirth; an increasing number of anomalies was more highly associated with stillbirth. Regardless of organ system affected, the presence of an anomaly increased the odds of stillbirth. These relationships remained significant if stillbirths with known genetic abnormalities were excluded. After multivariable analyses, the adjusted odds ratio (aOR) of stillbirth for any anomaly was 4.33 (95% CI 2.80-6.70) and the systems most strongly associated with stillbirth were cystic hygroma (aOR 29.97, 95% CI 5.85-153.57), and thoracic (aOR16.18, 95% CI 4.30-60.94) and craniofacial (aOR 35.25, 95% CI 9.22-134.68) systems. CONCLUSIONS: In pregnancies affected by anomalies, the odds of stillbirth are higher with increasing numbers of anomalies. Anomalies of nearly any organ system increased the odds of stillbirth even when adjusting for gestational age and maternal race. TWEETABLE ABSTRACT: Stillbirth risk increases with anomalies of nearly any organ system and with number of anomalies seen.

Mask-induced dermatoses during the COVID-19 pandemic: a questionnaire-based study in 12 Korean hospitals.

BACKGROUND: During the coronavirus disease 2019 (COVID-19) pandemic, various adverse skin reactions to long-term mask wearing have been reported. AIM: To assess the clinical features of mask-induced dermatoses and to recommend prevention and treatment options. METHODS: From April to August 2020, questionnaires including topics such as demographic information, pre-existing skin disorders, reported mask-related symptoms, daily mask-wearing duration and frequency, types of masks used and whether the participant was a healthcare worker, were distributed to patients in 12 hospitals. Dermatologists assessed skin lesions, confirmed diagnosis and recorded treatments. RESULTS: Itchiness was the most frequent symptom, mostly affecting the cheeks. The most common skin disease was new-onset contact dermatitis (33.94%), followed by new-onset acne (16.97%) and worsening of pre-existing acne (16.97%). Daily wearing of masks was significantly (P = 0.02) associated with new-onset contact dermatitis. More than half of patients with pre-existing skin problems experienced disease worsening while wearing masks. Longer duration of wearing (> 6 h/day, P = 0.04) and use of cotton masks (P < 0.001) significantly increased acne flare-up. Healthcare workers had a higher incidence of skin disease. Skin lesions were generally mild and well tolerated with topical treatment. The study had some limitations: the effect of seasonal characteristics and other risk factors were not assessed, and the patients were visiting dermatological clinics and had interest in their skin status, thus, there may have been selection bias. CONCLUSION: Mask-induced/-triggered dermatoses contribute to increase the dermatological burden during the pandemic.

Insight into the Chemistry of PETN Under Shock Compression Through Ultrafast Broadband Mid-Infrared Absorption Spectroscopy.

Thin films of pentaerythritol tetranitrate (PETN) were shock compressed using the laser driven shock apparatus at Los Alamos National Laboratory (LANL). Two spectroscopic probes were available to this apparatus: visible white light transient absorption spectroscopy (VIS) from 400 to 700 nm and mid-infrared transient absorption spectroscopy (MIR) from 1150 to 3800 cm-1. Important PETN vibrational modes are the symmetric and antisymmetric NO2 stretches at 1280 and 1650 cm-1, respectively, as well as CH stretches at ∼2900 cm-1. Shock strength was varied from approximately 3 to 55 GPa to span from the chemically unreactive regime to the regime in which fast chemical reaction took place on the 250 ps time scale of the measurements. VIS and MIR results suggest irreversible chemistry was induced in PETN at pressures above 30 GPa. At lower shock pressures, the spectroscopy showed minimal changes attributable to pressure induced effects. Under the higher-pressure reactive conditions, the frequency region at the antisymmetric NO2 stretch mode had a significantly increased absorption while the region around the symmetric NO2 stretch did not. No observable increased absorption occurred in the higher frequency regions where CH-, NH-, and OH- bond absorptions would be observed. A broad absorption appeared on the shoulder at the red-edge of the CO2 vibrational band around 2200 cm-1. In addition to the experiments, reactive molecular dynamics were carried out under equivalent shock conditions to correlate the evolution of the infrared spectrum to molecular processes. The simulations show results consistent to experiments up to 30 GPa but suggest that NO and NO2 related features provided the strongest contributions to the shocked infrared changes. Proposed mechanisms for shocked PETN chemistry are analyzed as consistent or inconsistent with the data presented here. Our experimental data suggests C≡O or N2O bond formation, nitrite formation, and absence of significant hydroxyl or amine concentrations in the initial chemistry steps in PETN shocked above 30 GPa.

Neuropathic pain in ankylosing spondylitis: a meta-analysis.

AIM: The aim of this study was to compare demographic characteristics, disease activity, functional status, and quality of life between ankylosing spondylitis (AS) with neuropatic pain (NP) and AS without NP (Non-NP). METHODS: The MEDLINE via PubMed, Cochrane, Scopus, and Embase database, from the earliest available date of indexing through December 20, 2018, were searched for comparative studies evaluating NP in AS patients. Two authors performed the data extraction independently. Any discrepancies were resolved by consensus. RESULTS: Four comparative studies were identified. There was no statistically significant difference in terms of age, body mass index, symptom duration, and inflammatory markers, such as erythrocyte sedimentation rate and C­reactive protein between NP and Non-NP. The sex ratios (F/M) were approximately 1/1 in NP and 1/2 in Non-NP and the proportion of human leukocyte antigen (HLA) B27-positive patients in NP and Non-NP was 65.7% and 83.0%, respectively. NP patients had significantly higher visual analogue scale pain scores, higher Bath Ankylosing Spondylitis Disease Activity Index, higher Bath Ankylosing Spondylitis Functional Index, and lower SF-Item Short Form physical component scores compare to Non-NP patients. CONCLUSION: The current meta-analysis showed that NP patients had significantly higher pain severity, higher disease activity and lower quality of life than Non-NP patients. The sex ratio (F/M) and proportion of HLA-B27 positive patients were different between the two groups. Further well-designed studies are needed to substantiate our results.

Silver nanoparticles induce Egr-1-dependent psoriasin expression via the ERK and p38 pathways.

BACKGROUND: Silver nanoparticles (Ag-NPs) can prevent bacterial infection and improve cutaneous wound healing owing to their antimicrobial activity. However, the mechanism of their antimicrobial activity is poorly understood. AIM: To determine the mechanistic relationship between Ag-NP treatment and expression of psoriasin. METHODS: Human epidermal keratinocytes, neonatal (HEKn) were used. Psoriasin mRNA expression was measured by reverse transcription PCR and real-time PCR. Western blotting was performed to verify expression of early growth response-1 (Egr-1) and psoriasin, and phosphorylation of mitogen-activated protein kinase (MAPK). Psoriasin promoter activity by Egr-1 was detected by a luciferase assay. RESULTS: Treatment of HEKn with Ag-NPs induced psoriasin mRNA and protein expression. Upregulation of psoriasin promoter activity was also observed in the luciferase assay. Ag-NPs increased Egr-1 expression, promoter activity and nuclear translocation in HEKn. Psoriasin luciferase activity was increased in HEKn transfected with Egr-1 pcDNA 3.1. Ag-NPs activated MAPK pathways including the extracellular signal-regulated kinase (ERK), p38, and c-Jun-N-terminal kinase (JNK) pathways. The upregulation of Egr-1 expression by Ag-NP stimulation was inhibited by ERK and p38 inhibitors, but not by a JNK inhibitor. Psoriasin expression was reduced in Egr-1 small interfering RNA-transfected HEKn. CONCLUSIONS: Ag-NP treatment induces upregulation of psoriasin expression through Egr-1 expression. We suggest that the ERK and p38 pathways are involved in Egr-1-dependent psoriasin expression.

Effect of the Lactobacillus rhamnosus strain GG and tagatose as a synbiotic combination in a dextran sulfate sodium-induced colitis murine model.

Synbiotics, a combination of prebiotics and probiotics, produce synergistic effects to promote gastrointestinal health. Herein, we investigated the synbiotic interaction between the Lactobacillus rhamnosus strain GG (LGG; a probiotic strain) and tagatose (a prebiotic) in a dextran sulfate sodium (DSS)-induced colitis murine model. Initially, body weight, food intake, and clinical features were dramatically decreased after treatment with DSS, and the addition of LGG, tagatose, or both ameliorated these effects. In our pyrosequencing analysis of fecal microbiota, DSS treatment increased the abundance of Proteobacteria and decreased that of Firmicutes. When LGG and tagatose were administered as synbiotics, the gut microbiota composition recovered from the dysbiosis caused by DSS treatment. In particular, the abundance of Bacteroides, Lactobacillus, and Akkermansia was significantly associated with probiotic, prebiotic, and synbiotic treatments. Taken together, our results suggest that LGG and tagatose as synbiotics can alleviate colitis, and synbiotics could be applied as dietary supplements in dairy foods such as yogurt and cheese.

Nanoporous anodic aluminum oxide internalized with gold nanoparticles for on-chip PCR and direct detection by surface-enhanced Raman scattering.

Nanoporous anodic aluminum oxide internalized with gold nanoparticles was utilized as an integrated platform miniaturized for consecutively performing on-chip PCR and downstream detection of the amplified product of a 183 bp eaeA gene fragment from Escherichia coli O157:H7 using surface-enhanced Raman scattering (SERS).

Efficacy of Sensory Transcutaneous Electrical Nerve Stimulation on Perceived Pain and Gait Patterns in Individuals With Experimental Knee Pain.

OBJECTIVES: To examine the effect of experimental knee pain on perceived knee pain and gait patterns and to examine the efficacy of transcutaneous electrical nerve stimulation (TENS) on perceived knee pain and pain-induced knee gait mechanics. DESIGN: Crossover trial. SETTING: Biomechanics laboratory. PARTICIPANTS: Recreationally active, individuals without musculoskeletal pain aged 18 to 35 years (N=30). INTERVENTIONS: Thirty able-bodied individuals were assigned to either a TENS (n=15) or a placebo (n=15) group. All participants completed 3 experimental sessions in a counterbalanced order separated by 2 days: (1) hypertonic saline infusion (5% NaCl); (2) isotonic saline infusion (0.9% NaCl); and (3) control. Each group received sensory electrical stimulation or placebo treatment for 20 minutes, respectively. MAIN OUTCOME MEASURES: Perceived pain was collected every 2 minutes using a 10-cm visual analog scale (VAS) for 50 minutes and analyzed using a mixed model analysis of covariance with repeated measures. Gait analyses were performed at baseline, infusion, and treatment. Sagittal and frontal knee angles and internal net joint torque across the entire stance were analyzed using a functional data analysis approach. RESULTS: Hypertonic saline infusion increased perceived pain (4/10cm on a VAS; P<.05) and altered right knee angle (more flexion and less abduction; P<.05) and internal net joint torque (less extension and greater abduction; P<.05) across various stance phases. TENS treatment reduced perceived pain and improved right sagittal gait abnormalities as compared with placebo treatment (P<.05). CONCLUSIONS: This pain model increases perceived pain and induces compensatory gait patterns in a way that indicates potential quadriceps weakness. However, TENS treatment effectively reduces perceived pain and restores pain-induced gait abnormalities in sagittal knee mechanics.

Effects of transcutaneous electrical nerve stimulation on quadriceps function in individuals with experimental knee pain.

Knee joint pain (KJP) is a cardinal symptom in knee pathologies, and quadriceps inhibition is commonly observed among KJP patients. Previously, KJP independently reduced quadriceps strength and activation. However, it remains unknown how disinhibitory transcutaneous electrical nerve stimulation (TENS) will affect inhibited quadriceps motor function. This study aimed at examining changes in quadriceps maximum voluntary contraction (MVC) and central activation ratio (CAR) before and after sensory TENS following experimental knee pain. Thirty healthy participants were assigned to either the TENS or placebo groups. All participants underwent three separate data collection sessions consisting of two saline infusions and one no infusion control in a crossover design. TENS or placebo treatment was administered to each group for 20 min. Quadriceps MVC and CAR were measured at baseline, infusion, treatment, and post-treatment. Perceived knee pain intensity was measured on a 100-mm visual analogue scale. Post-hoc analysis revealed that hypertonic saline infusion significantly reduced the quadriceps MVC and CAR compared with control sessions (P < 0.05). Sensory TENS, however, significantly restored inhibited quadriceps motor function compared with placebo treatment (P < 0.05). There was a negative correlation between changes in MVC and knee pain (r = 0.33, P < 0.001), and CAR and knee pain (r = 0.62, P < 0.001), respectively.

Short communication: In vivo screening platform for bacteriocins using Caenorhabditis elegans to control mastitis-causing pathogens.

This study aimed to develop an in vivo screening platform using Caenorhabditis elegans to identify a novel bacteriocin for controlling the mastitis-causing pathogen Staphylococcus aureus strain RF122 in dairy cows. Using Bacillus spp. isolated from traditional Korean foods, we developed a direct in vivo screening platform that uses 96-well plates and fluorescence image analysis. We identified a novel bacteriocin produced by Bacillus licheniformis strain 146 (lichenicin 146) with a high in vivo antimicrobial activity using our liquid C. elegans-Staph. aureus assay. We also determined the characteristics of lichenicin 146 using liquid chromatography-mass spectrometry and confirmed that it shared homologous sequences with bacteriocin family proteins. In addition, RNA-sequencing analysis revealed genes encoding cell surface or membrane proteins (SAB0993c, SAB0150, SAB0994c, and SAB2375c) that are involved in the bactericidal activity of lichenicin 146 against Staph. aureus strain RF122 infection as well as those encoding transcriptional regulators (SAB0844c and SAB0133). Thus, our direct in vivo screening platform facilitates simple, convenient, cost-effective, and reliable screening of potential antimicrobial compounds with applications in the dairy field.

Non-invasive assessment of cutaneous wound healing using fluorescent imaging.

BACKGROUND/PURPOSE: Optical imaging is a very important technique in the biomedical sciences. The purpose of this study was to develop an in vivo optical system for fluorescent imaging and molecular imaging applications using quantum dots (QDs). METHODS: The in vivo optical system was composed of modular parts, including a light source, light guide, excitation filter wheel, excitation filters, emission filter wheel, emission filters, liquid crystal tunable filter (LCTF), macro lens, dark chamber, and a cooled charged-coupled device (CCD) camera for recording images. Filters were selected based on the excitation and absorption spectra of QDs to allow spectral separation and optimization of the acquired image. In contrast with conventional systems, our system allows selection of the emission bandwidth. RESULTS: The system was tested in an in vivo study using a wound-healing model in nude mice. The healing process was examined after injection of fibroblasts and keratinocytes labeled with two different sets of QDs. The different QD probes were readily detected and distinguished using our system. CONCLUSION: An in vivo optical system is a very useful tool for the detection of genes, proteins, and small-molecule drugs inside living animals, and this imaging modality can also be adopted for real-time visualization of cancer cell metastasis in live animals.

Functional Fatigue Alters Lower-extremity Neuromechanics during a Forward-side Jump.

Neuromuscular fatigue impairs neuromuscular control of the lower extremity. The purpose of the study was to investigate the effect of functional fatiguing exercises on sagittal-plane lower-extremity neuromechanics during a forward-side jump. 21 participants performed 5 forward-side jump tasks before and after functional fatiguing exercises. A functional analysis of variance (FANOVA) evaluated differences between 2 different conditions (pre- vs. post-fatigue) for joint angle, moment, and EMG amplitude during stance of a forward-side jump. FANOVA compared variables as polynomial functions, and differences between functions with 95% confidence interval bands were plotted to determine significant differences. Plantar, knee, and hip flexion decreased during the initial stages of landing following fatigue. Plantarflexion moment decreased during 10-20% of stance in post-fatigue. Knee extension moment initially increased while decreased during 20-30% of stance following fatigue. Hip extension moment initially decreased while increased at 20% of stance. Tibialis anterior EMG decreased during 30-40% of stance, vastus lateralis EMG increased at 15% of stance, hamstring EMG decreased at foot contact and during 25-60% of stance, and gluteus maximus EMG decreased at foot contact and 35% of stance. The functional fatiguing exercises resulted in a more upright landing position, potentially indicating a greater reliance upon skeletal structures for support.

Indole derivatives inhibit hepatitis C virus replication through induction of pro-inflammatory cytokines.

Previously, we discovered a series of indole derivatives as a new class of hepatitis C virus (HCV) replication inhibitors by using a target-free chemical genetic strategy. Through a structure-activity relationship study, the compound 12e was identified as the most potent inhibitor of this class (EC50 = 1.1 µmol/l) with minimal cytotoxicity (CC50 = 61.8 µmol/l). In order to gain insight into its detailed antiviral mechanism of action, we performed PCR array analyses and found that 12e was able to activate transcription of a number of pro-inflammatory as well as antiviral cytokine genes including CXCL-8, IL-1α, TNF-α, IL-3, IRAK-1, and DDX58. Their induction by 12e was verified by individual RT-PCR analyses. In addition, 12e was found to stimulate secretion of soluble factors with anti-HCV replication activity. Among the 12e-induced pro-inflammatory cytokines, CXCL-8 showed a strong positive correlation between its transcriptional activation and antiviral potency. Interestingly, a recombinant CXCL-8 protein also reduced HCV replication, though only moderately. In conclusion, we found a novel mode of action of indole derivatives in inhibiting HCV replication, particularly the induction of pro-inflammatory cytokines.

Total Chemical Synthesis of an Orf Virus Protein, ORFV002, an Inhibitor of the Master Gene Regulator NF-κB.

ORFV002 is a novel orf viral protein (117 Aa) that inhibits nuclear events through the regulation of the transcriptional activity of NF-κB, a master regulator of human gene expression (Diel et al., J Virol 2011, 85, 264-275). It is identified as the first nuclear inhibitor of NF-κB produced by orf virus (ORFV) and no homologues in other genera of the Chordopoxvirinae subfamily have been reported to date (Diel et al., J Virol 2011, 85, 264-275). Our molecular structure predictions suggest that ORFV002 may mimic part of IκB, an inhibitor and natural human partner of NF-κB. Recent advances in total chemical synthesis of proteins have provided solutions in overcoming challenges of current recombinant methods of protein isolation for structure elucidation. Aided by Boc solid phase peptide synthesis and native chemical ligation, ORFV002 was successfully synthesized in multimilligram amounts in good yield and high purity.