Multi-detector fusion and Bayesian smoothing for tracking viral and chromatin structures.

Automatic tracking of viral and intracellular structures displayed as spots with varying sizes in fluorescence microscopy images is an important task to quantify cellular processes. We propose a novel probabilistic tracking approach for multiple particle tracking based on multi-detector and multi-scale data fusion as well as Bayesian smoothing. The approach integrates results from multiple detectors using a novel intensity-based covariance intersection method which takes into account information about the image intensities, positions, and uncertainties. The method ensures a consistent estimate of multiple fused particle detections and does not require an optimization step. Our probabilistic tracking approach performs data fusion of detections from classical and deep learning methods as well as exploits single-scale and multi-scale detections. In addition, we use Bayesian smoothing to fuse information of predictions from both past and future time points. We evaluated our approach using image data of the Particle Tracking Challenge and achieved state-of-the-art results or outperformed previous methods. Our method was also assessed on challenging live cell fluorescence microscopy image data of viral and cellular proteins expressed in hepatitis C virus-infected cells and chromatin structures in non-infected cells, acquired at different spatial-temporal resolutions. We found that the proposed approach outperforms existing methods.

Mental health impact of multiple sexually minoritized and gender expansive stressors among LGBTQ+ young adults: a latent class analysis.

AIMS: In the United States, lesbian, gay, bisexual, transgender, queer, intersex, asexual and other sexually minoritized and gender expansive (LGBTQ+) young adults are at increased risk for experiencing mental health inequities, including anxiety, depression and psychological distress-related challenges associated with their sexual and gender identities. LGBTQ+ young adults may have unique experiences of sexual and gender minority-related vulnerability because of LGBTQ+-related minority stress and stressors, such as heterosexism, family rejection, identity concealment and internalized homophobia. Identifying and understanding specific LGBTQ+-related minority stress experiences and their complex roles in contributing to mental health burden among LGBTQ+ young adults could inform public health efforts to eliminate mental health inequities experienced by LGBTQ+ young adults. Therefore, this study sought to form empirically based risk profiles (i.e., latent classes) of LGBTQ+ young adults based on their experiences with familial heterosexist experiences, LGBTQ+-related family rejection, internalized LGBTQ+-phobia and LGBTQ+ identity concealment, and then identify associations of derived classes with psychological distress. METHODS: We recruited and enrolled participants using nonprobability, cross-sectional online survey data collected between May and August 2020 (N = 482). We used a three-step latent class analysis (LCA) approach to identify unique classes of response patterns to LGBTQ+-related minority stressor subscale items (i.e., familial heterosexist experiences, LGBTQ+-related family rejection, internalized LGBTQ+-phobia and LGBTQ+ identity concealment), and multinomial logistic regression to characterize the associations between the derived classes and psychological distress. RESULTS: Five distinct latent classes emerged from the LCA: (1) low minority stress, (2) LGBTQ+ identity concealment, (3) family rejection, (4) moderate minority stress and (5) high minority stress. Participants who were classified in the high and moderate minority stress classes were more likely to suffer from moderate and severe psychological distress compared to those classified in the low minority stress class. Additionally, relative to those in the low minority stress class, participants who were classified in the LGBTQ+ identity concealment group were more likely to suffer from severe psychological distress. CONCLUSION: Familial heterosexist experiences, LGBTQ+-related family rejection, internalized LGBTQ+-phobia and LGBTQ+ identity concealment are four constructs that have been extensively examined as predictors for mental health outcomes among LGBTQ+ persons, and our study is among the first to reveal nuanced gradients of these stressors. Additionally, we found that more severe endorsement of minority stress was associated with greater psychological distress. Given our study results and the previously established negative mental health impacts of minority stressors among LGBTQ+ young adults, findings from our study can inform research, practice, and policy reform and development that could prevent and reduce mental health inequities among LGBTQ+ young adults.

An Understated Comorbidity: The Impact of Homelessness on Traumatic Brain Injury.

Traumatic brain injury (TBI), a neurovascular injury caused by external force, is a common diagnosis among veterans and those experiencing homelessness (HL). There is a significant overlap in the veteran and homeless population, possibly accounting for the two to seven times greater incidence of TBI among those experiencing HL than the general population. Despite these statistics, individuals experiencing HL are often underdiagnosed and ineffectively treated for TBI. We introduced a novel model of HL. Over 5 weeks, adult Sprague-Dawley rats were randomly assigned to one of the following conditions: TBI only, HL only, TBI + HL, or control (n = 9 per group). To emulate HL, animals (2 animals per cage) were exposed to soiled beddings for 5 weeks. Subsequently, animals were introduced to TBI by using the moderate controlled cortical impact model, then underwent 4 consecutive days of behavioral testing (beam walk (BW), elevated body swing test (EBST), forelimb akinesia (FA), paw grasp (PG), Rotorod, and elevated T-maze). Nissl staining was performed to determine the peri-impact cell survival and the integrity of corpus callosum area. Motor function was significantly impaired by TBI, regardless of housing (beam walk or BW 85.0%, forelimb akinesia or FA 104.7%, and paw grasp or PG 100% greater deficit compared to control). Deficits were worsened by HL in TBI rats (BW 93.3%, FA 40.5%, and PG 50% greater deficit). Two-way ANOVA revealed BW (F(4, 160) = 31.69, p < 0.0001), FA (F(4, 160) = 13.71, p < 0.0001), PG (F(4, 160) = 3.873, p = 0.005), Rotorod (F(4, 160), p = 1.116), and EBST (F(4, 160) = 6.929, p < 0.0001) showed significant differences between groups. The Rotorod and EBST tests showed TBI-induced functional deficits when analyzed by day, but these deficits were not exacerbated by HL. TBI only and TBI + HL rats exhibited typical cortical impact damage (F(3,95) = 51.75, p < 0.0001) and peri-impact cell loss compared to control group (F(3,238) = 47.34, p < 0.0001). Most notably, TBI + HL rats showed significant alterations in WM area measured via the corpus callosum (F(3, 95) = 3.764, p = 0.0133). Worsened behavioral outcomes displayed by TBI + HL rats compared to TBI alone suggest HL contributes to TBI functional deficits. While an intact white matter, such as the corpus callosum, may lessen the consequent functional deficits associated with TBI by enhancing hemispheric communications, there are likely alternative cellular and molecular pathways mitigating TBI-associated inflammatory or oxidative stress responses. Here, we showed that the environmental condition of the patient, i.e., HL, participates in white matter integrity and behavioral outcomes, suggesting its key role in the disease diagnosis to aptly treat TBI patients.

Regional and annual patterns in respiratory virus co-infection etiologies and antibiotic prescriptions for pediatric mycoplasma pneumoniae pneumonia.

OBJECTIVE: Mycoplasma pneumoniae (M. pneumoniae) pneumonia is the second-most common cause of community-acquired pneumonia (CAP). This study aimed at investigating into the prevalence of macrolide-resistant M. pneumoniae (MRMP) with respiratory virus co-infection and the antibiotic prescriptions in children with CAP in four provinces in Korea, and to assess the variations in the findings across regions and throughout the year. PATIENTS AND METHODS: This prospective study was conducted in 29 hospitals in Korea between July 2018 and June 2020. Among the enrolled 1,063 children with CAP, all 451 patients with M. pneumoniae underwent PCR assays of M. pneumoniae and respiratory viruses, and the presence of point mutations of residues 2063 and 2064 was evaluated. RESULTS: Gwangju-Honam (88.6%) showed the highest prevalence of MRMP pneumonia, while Daejeon-Chungcheong (71.3%) showed the lowest, although the differences in prevalence were not significant (p=0.074). Co-infection of M. pneumoniae pneumonia and respiratory virus was observed in 206 patients (45.4%), and rhinovirus co-infection (101 children; 22.2%) was the most frequent. The prevalence of MRMP pneumonia with respiratory virus co-infection and the antibiotic prescriptions differed significantly among the four provinces (p < 0.05). The monthly rate of MRMP pneumonia cases among all cases of M. pneumoniae pneumonia and tetracycline or quinolone prescriptions did not differ significantly among the four regions (trend p > 0.05) during the study period. CONCLUSIONS: The prevalence of M. pneumoniae pneumonia with virus co-infection and antibiotic prescriptions could differ according to region, although the MRMP pneumonia rate showed no difference within Korea.

Identification of new IL-7Rα small-molecule agonists: a multi-computational approach.

Interleukin 7 (IL-7) is an essential cytokine that acts as a potent growth factor of T-cells and supports the growth of B-cell precursors. IL-7 binds to a heterodimeric receptor consisting of an IL-7 receptor alpha (IL-7Rα) and the common gamma chain receptor (γc) which is shared with IL-2, IL-4, IL-9, IL-15 and IL-21. The discovery of small-molecule agonists of cytokines would be of great pharmaceutical interest with the increasing scientific rationale. In this study, a series of molecular modelling methods, including field-based pharmacophore virtual screening, protein-protein docking and molecular dynamics simulations, led to the identification of two compounds (i.e. 1 and 2) of different classes that exhibit enhanced agonistic effects by activating the IL-7 signalling cascade. One of these compounds was selected as a hit and represents the first small-molecule agonist of IL-7Rα with single-digit micromolar activity. Moreover, the prediction model of the active compound to the IL-7Rα/γc interaction complex provides insight into the binding of a small-molecule agonist to its receptor.

Data fusion and smoothing for probabilistic tracking of viral structures in fluorescence microscopy images.

Automatic tracking of viral structures displayed as small spots in fluorescence microscopy images is an important task to determine quantitative information about cellular processes. We introduce a novel probabilistic approach for tracking multiple particles based on multi-sensor data fusion and Bayesian smoothing methods. The approach exploits multiple measurements as in a particle filter, both detection-based measurements and prediction-based measurements from a Kalman filter using probabilistic data association with elliptical sampling. Compared to previous probabilistic tracking methods, our approach exploits separate uncertainties for the detection-based and prediction-based measurements, and integrates them by a sequential multi-sensor data fusion method. In addition, information from both past and future time points is taken into account by a Bayesian smoothing method in conjunction with the covariance intersection algorithm for data fusion. Also, motion information based on displacements is used to improve correspondence finding. Our approach has been evaluated on data of the Particle Tracking Challenge and yielded state-of-the-art results or outperformed previous approaches. We also applied our approach to challenging time-lapse fluorescence microscopy data of human immunodeficiency virus type 1 and hepatitis C virus proteins acquired with different types of microscopes and spatial-temporal resolutions. It turned out, that our approach outperforms existing methods.

Chronic rhinosinusitis and progression of cognitive impairment in dementia.

AIM: The relationship between chronic inflammatory disease and cognitive decline is still unclear, but there is increasing evidence to support the role of systemic inflammation. The aim of this study was to investigate if chronic rhinosinusitis (CRS) in dementia or mild cognitive impairment (MCI) is associated with the progression of cognitive decline. MATERIAL AND METHODS: We retrospectively reviewed the data of patients who complained of memory impairment, and underwent brain magnetic resonance imaging (MRI) from January 2006 to April 2019. According to the Mini-Mental State Examination (MMSE) score, subjects (n=661) were divided into three groups: dementia (≤17), MCI (18-23), and normal (≥24). CRS was defined as a total score of greater than or equal to 4 according to the Lund-Mackay scoring system using brain MRI. Multiple logistic regression analyses estimated adjusted odds ratio (aOR) for the association between CRS and dementia or MCI. Among the subjects with follow-up MMSE (n=286), a repeated-measures ANOVA was used to assess the difference of changes in MMSE scores between the groups with and without CRS. RESULTS: According to the initial MMSE score, there were 221 subjects with dementia, 195 with MCI, and 245 with normal results. CRS was not significantly associated with dementia (aOR=1.519, CI=0.909-2.538, P=0.111), while being suggestively associated with MCI (aOR=1.740, CI=1.041-2.906, P=0.034). The MMSE scores at follow-up decreased further in subjects with CRS than in those without CRS (P=0.009). Especially, in the initial dementia group, there was a significant between-group difference in the MMSE score from baseline to follow-up (13.6±4.3 to 11.1±6.3 in CRS group vs. 13.5±3.3 to 14.4±5.4 in no CRS group, P=0.002). CONCLUSION: The result of the present study implies a potential association between CRS and progression of cognitive decline. Physicians should be aware of this possibility in patients with clinically diagnosed CRS.

Effects of High-Speed Power Training on Neuromuscular and Gait Functions in Frail Elderly with Mild Cognitive Impairment Despite Blunted Executive Functions: A Randomized Controlled Trial.

BACKGROUND: Physical frailty and impaired executive function of the brain show similar pathophysiology. Both of these factors lead to dysfunction of neuromuscular and abilities in elderly. High-speed power training (HSPT) has been determined to have positive effects on neuromuscular function and gait performance, as well as executive function in the elderly. OBJECTIVES: The purpose of this study was to investigate the effects of 8-week HSPT on neuromuscular, gait and executive functions in frail elderly with mild cognitive impairment (MCI). DESIGN, SETTING AND PARTICIPANTS: We performed a randomized controlled trial of frail elderly from community and medical center in republic of Korea. Forty-two physically frail elderly with MCI were randomly allocated to control (n=22, age=74.22±4.46) and intervention groups (n=18, age=73.77±4.64). The intervention group was subjected to HSPT, 3 times weekly for 8 weeks. MEASUREMENTS: Isometric contraction of knee extension and flexion with electromyography (EMG) was measured to determine the neuromuscular function such as knee extensor strength, rate of torque development, movement time, pre-motor time, motor time, rate of EMG rise, and hamstrings antagonist co-activation. Additionally, the 4.44-meter gait and timed up-and-go (TUG) test were administered to assess gait performance. A frontal assessment battery was measured in this study. RESULTS: The 8-week HSPT regimen improved the knee extensor strength from 1.13±0.08 to 1.25±0.07 (p<0.05), the 200-ms RTD from 3.01±0.3 to 3.55±0.24 (p<0.05) and the rate of EMG rise from 166.48±13.31 to 197.94±11.51 (p<0.05), whereas the movement time and motor time were statistically decreased from 921.69±40.10 to 799.51±72.84, and 271.40±19.29 to 181.15±38.08 (p<0.05), respectively. The 4.44-m gait speed and TUG significantly decreased from 6.39±0.25 to 5.5±0.24, and 11.05±0.53 to 9.17±0.43 respectively (p<0.05). CONCLUSION: The findings of this study suggest the favorable effects of 8-week HSPT on the neuromuscular function and the gait performance in the frail elderly with MCI without increase in the executive function.

Survival of oral mucosal melanoma according to treatment, tumour resection margin, and metastases.

Because of the poor prognosis and of oral mucosal melanoma, and patients' short survival, large, randomised, clinical studies are difficult. We have investigated its demographic characteristics and analysed the effect of treatment, resection margins, and metastases on survival. We recorded age, sex, site of primary tumour, and types of treatment, survival, and metastases in 74 patients treated at the Department of Oral and Maxillofacial Surgery, Seoul National University Dental Hospital. Survival was analysed based on bony invasion, depth of invasion, and resection margins, and we found that it varied depending on the primary site (p=0.002), and declined with liver (p=0.001) or brain (p=0.033) metastases. The two-year survival according to the primary site was as follows: palate 85% (n=32), anterior maxillary gingiva 53% (n=13), mandible 58% (n=13), and posterior maxillary gingival 74% (n=10) and buccal mucosa 50% (n=4). The two-year survival was 34% (n=8) in patients with liver metastases and 23% (n=7) in patients with brain metastases. In cases of bony invasion (p=0.005), depth of invasion (p=0.042), unclear resection margin (p=0.023), or higher T stages (p=0.009), the survival declined considerably. Neck dissection did not affect survival (p=0.343). Survival of the patients given chemotherapy was significantly lower (p=0.013) and the two-year survival was 54.0%. The patients given radiotherapy showed no significant difference in survival compared with those not given radiotherapy (p=0.107). In conclusion, primary site, bony invasion, resection margins, depth of invasion and systemic metastases were critical to predict prognosis and selection of treatment of oral mucosal melanoma.

Outbreak of carbapenemase-producing Enterobacteriaceae associated with a contaminated water dispenser and sink drains in the cardiology units of a Korean hospital.

BACKGROUND: Concerns are growing over the importance of the hospital water environment for the transmission of carbapenemase-producing Enterobacteriaceae (CPE). AIM: To report a large outbreak in the cardiology units involving intensive care units (ICUs) and wards at a tertiary-care hospital. METHODS: This was a contact tracing, case-control study to find the risk factors for acquisition of CPE and environmental sampling was performed during a CPE outbreak between July and December 2018. FINDINGS: A total of 87 patients with CPE infection or colonization were identified in the cardiology units of the Asan Medical Centre. Diverse organisms were identified containing blakpc, blaNDM-1, blaVIM or blaIMP, blaOXA-48, and co-producing organisms. A case-control study indicated that using the sinks in the ward patient room bathroom for teeth brushing was associated with CPE acquisition (83% vs 30%; P=0.03). The environment was cultured and Klebsiella pneumoniae carbapenemase (KPC)-producing Escherichia coli was isolated from a water dispenser and New Delhi metallo-beta-lactamase (NDM) 1-producing Citrobacter freundii and Enterobacter cloacae from sinks in patient rooms. Pulsed-field gel electrophoresis (PFGE) analysis of KPC-producing E. coli from patients and the water dispenser in ICU and NDM-1-producing E. cloacae from the patient and sink drain showed the same pulsotypes. CONCLUSIONS: The water dispenser and sink drain were suspected as possible reservoirs of CPE in this outbreak. Close contacts with contaminated water such as tooth brushing were identified as risk factors for CPE acquisition. Education for the adequate use of the water environment system as well as the control of the hospital water environment should be implemented to prevent the CPE outbreaks.

Airway changes and prevalence of obstructive sleep apnoea after bimaxillary orthognathic surgery with large mandibular setback.

This study used three-dimensional computed tomography and polysomnography to evaluate the effect of a large mandibular setback on the postoperative pharyngeal airway space and obstructive sleep apnoea (OSA). Twelve patients who underwent bimaxillary surgery for a mandibular setback movement of >9mm were included in this study. Changes in the pharyngeal airway space and polysomnography parameters based on the surgical movements were analyzed. The median mandibular setback movement was 11.08mm. The total pharyngeal, oropharyngeal, and hypopharyngeal volumes, and the retroglossal cross-sectional area were significantly decreased postoperatively (P=0.006; P=0.005; P=0.012; P=0.005, respectively). The apnoea-hypopnoea index (AHI) increased significantly after surgery (P=0.021). There were significant positive correlations between the preoperative inferiorly located hyoid bone and both AHI and respiratory disturbance index (RDI) postoperative (P=0.008 and P=0.027) and between the postoperative inferiorly dislocated retropalatal level and both AHI and RDI postoperative (P=0.002 and P=0.014). Four patients (33.3%) developed new onset OSA postoperatively. Large mandibular setback movements significantly reduced the pharyngeal airway space in the setting of bimaxillary surgery (P=0.006).

Overall survival results of AGO-OVAR16: A phase 3 study of maintenance pazopanib versus placebo in women who have not progressed after first-line chemotherapy for advanced ovarian cancer.

OBJECTIVE: The AGO-OVAR16 study was designed to test the efficacy, safety, and tolerability of pazopanib maintenance after first-line chemotherapy in patients with newly diagnosed advanced ovarian cancer (AOC). METHODS: Nine hundred and forty patients with histologically confirmed AOC, International Federation of Gynecology and Obstetrics (FIGO) stage II-IV, were randomized in a 1:1 ratio to receive either 800 mg pazopanib once daily or placebo for up to 24 months, unless there was disease progression, toxicity, withdrawal of consent, or death. The primary endpoint (investigator-assessed progression-free survival [PFS]) was met and previously reported. The results of final analyses of overall survival (OS) are reported here. RESULTS: A third OS interim analysis showed futility and led to study closure and a final OS analysis after last patient last visit. At the time of the final OS analysis, 494 (89.7% of the planned 551) events had occurred. No difference was observed in OS between pazopanib and placebo. The hazard ratio (HR) was 0.960 (95% confidence interval [CI]: 0.805-1.145), and the median OS from randomization was 59.1 months in pazopanib and 64.0 months in placebo arms. For the East Asian patients, similar to the first three interim OS analyses, a numerical negative trend was observed favoring placebo (HR, 1.332; 95% CI: 0.863-2.054). Exploratory analyses showed a trend for a longer time to first subsequent anti-cancer therapy or death with pazopanib over placebo (HR, 0.829; 95% CI: 0.713-0.965), with a median estimate of 19.0 and 14.5 months, respectively. No new safety signals were observed. CONCLUSION: Although pazopanib prolonged PFS, this was not associated with improvement in median OS. CLINICAL TRIAL INFORMATION: ClinicalTrials.gov: NCT00866697.

North American April tornado occurrences linked to global sea surface temperature anomalies.

Annual tornado occurrences over North America display large interannual variability and a statistical linkage to sea surface temperature (SST) anomalies. However, the underlying physical mechanisms for this connection and its modulation in a rapidly varying seasonal environment still remain elusive. Using tornado data over the United States from 1954 to 2016 in combination with SST-forced atmospheric general circulation models, we show a robust dynamical linkage between global SST conditions in April, the emergence of the Pacific-North American teleconnection pattern (PNA), and the year-to-year tornado activity in the Southern Great Plains (SGP) region of the United States. Contrasting previous studies, we find that only in April SST-driven atmospheric circulation anomalies can effectively control the northward moisture-laden flow from the Gulf of Mexico, boosting low-level moisture flux convergence over the SGP. These strong large-scale connections are absent in other months because of the strong seasonality of the PNA and background moisture conditions.

A novel method to test non-exclusive hypotheses applied to Arctic ice projections from dependent models.

A major conundrum in climate science is how to account for dependence between climate models. This complicates interpretation of probabilistic projections derived from such models. Here we show that this problem can be addressed using a novel method to test multiple non-exclusive hypotheses, and to make predictions under such hypotheses. We apply the method to probabilistically estimate the level of global warming needed for a September ice-free Arctic, using an ensemble of historical and representative concentration pathway 8.5 emissions scenario climate model runs. We show that not accounting for model dependence can lead to biased projections. Incorporating more constraints on models may minimize the impact of neglecting model non-exclusivity. Most likely, September Arctic sea ice will effectively disappear at between approximately 2 and 2.5 K of global warming. Yet, limiting the warming to 1.5 K under the Paris agreement may not be sufficient to prevent the ice-free Arctic.

Cognitive decline in association with hyposmia in idiopathic rapid eye movement sleep behavior disorder: a prospective 2-year follow-up study.

BACKGROUND AND PURPOSE: The aim was to analyze the characteristics and progression of cognitive dysfunction in non-demented idiopathic rapid eye movement sleep behavior disorder (iRBD) patients with baseline olfactory function. METHODS: From a prospective polysomnography-confirmed iRBD cohort, 25 patients (16 patients in 2-year follow-up) and 13 normal controls were included. Initial and 2-year follow-up cognitive functions were analyzed with olfactory function and 18 F-fluorinated-N-3-fluoropropyl-2ß-carboxymethoxy-3ß-(4-iodophenyl)-nortropane (18 F-FP-CIT) uptake in deep nuclei initially. RESULTS: Idiopathic RBD patients had impaired attention, memory and executive function compared to controls. Baseline cognitive tests were comparable between the iRBD subgroups with and without hyposmia. 18 F-FP-CIT uptake tended to be lower in the hyposmic group than in the normosmic group. The olfactory test score was positively correlated with amygdala uptake in iRBD patients (P = 0.027). After 2 years, visuospatial and verbal memory dysfunction worsened more in hyposmics than in normosmics. Lower initial olfactory test score was associated with more severe declines in verbal memory function. CONCLUSIONS: Hyposmia may be a predictive sign of cognitive decline in iRBD patients.

Lytic transglycosylase contributes to the survival of lipooligosaccharide-deficient, colistin-dependent Acinetobacter baumannii.

OBJECTIVES: The phenomenon of colistin dependence in Acinetobacter baumannii has been described in a situation in which colistin is now considered as the last resort for the treatment of infections caused by multidrug-resistant Gram-negative bacteria. In this study, we aimed to reveal a gene associated with colistin dependence in A. baumannii. METHODS: The colistin-dependent A. baumannii H08-391D strain was isolated from a patient, and target gene-inactivation mutants were constructed. We investigated the effects of target gene on colistin dependence with quantitative real-time PCR and endotoxin assay. Also, we observed the change of cell morphology by electron microscopy. RESULTS: The expression of ACICU_02898, encoding a soluble lytic transglycosylase associated with cell-wall degradation and recycling, was increased by eight-to 42-fold in colistin-dependent mutants, and deletion of ACICU_02898 in a colistin-dependent strain led to colistin susceptibility (MIC = 8 mg/L). Endotoxin activity was significantly low in a colistin-dependent derivative ACICU_02898-inactivated mutant and a complemented mutant. In addition, the ACICU_02898-inactivated mutant showed a highly reduced growth rate. The colistin-dependent derivative and ACICU_02898-inactivated mutant showed clearly distinguished absorption profiles in the red/green fluorescence dot blot with regard to their membrane potential. Electron microscopy revealed that the deletion mutant cells were elongated compared to the colistin-susceptible wild-type strain and colistin-dependent strain. CONCLUSIONS: A colistin-dependent A. baumannii strain exhibited a deficiency in its outer membrane integrity and high expression of lytic transglycosylase was required for survival. This study reveals why the colistin-dependent mutant can tolerate high antibiotic concentrations.

Structural and molecular modelling studies of antimelanogenic piper-amide TRPM1 antagonists.

Piper-amides exhibit diverse biological activities, including antimelanogenic effects. In our previous studies, we identified a potent piper-amide derivative that inhibited melanogenesis via the TRPM1 calcium channel. Despite its potential as a therapeutic target, the three-dimensional structure of TRPM1 is still not available. Thus, structure-guided compound design and the discovery of novel inhibitors of melanogenesis have been limited. In the present study, a series of computational methods, including homology modelling, docking, molecular dynamics simulation and field-based pharmacophore modelling, were integrated to explore the structural features of natural piper-amide-like compounds related to the TRPM1 target. These studies suggested the binding mode and provided a 3D pharmacophore model of the ligands, which can be helpful in understanding the TRPM1-ligand interactions at the molecular level and in designing potent antagonists of TRPM1.