Association of systemic secondary brain insults and outcome in patients with convulsive status epilepticus: A post hoc study of a randomized controlled trial.
To evaluate the association between systemic factors (mean arterial blood pressure, arterial partial pressures of carbon dioxide and oxygen, body temperature, natremia, and glycemia) on day 1 and neurologic outcomes 90 days after convulsive status epilepticus. This was a post hoc analysis of the Evaluation of Therapeutic Hypothermia in Convulsive Status Epilepticus in Adults in Intensive Care (HYBERNATUS) multicenter open-label controlled trial, which randomized 270 critically ill patients with convulsive status epilepticus requiring mechanical ventilation to therapeutic hypothermia (32°C-34°C for 24 hours) plus standard care or standard care alone between March 2011 and January 2015. The primary endpoint was a Glasgow Outcome Scale score of 5, defining a favorable outcome, 90 days after convulsive status epilepticus. The 172 men and 93 women had a median age of 57 years (45-68 years). Among them, 130 (49%) had a history of epilepsy, and 59 (29%) had a primary brain insult. Convulsive status epilepticus was refractory in 86 (32%) patients, and total seizure duration was 67 minutes (35-120 minutes). The 90-day outcome was unfavorable in 126 (48%) patients. In multivariate analysis, none of the systemic secondary brain insults were associated with outcome; achieving an unfavorable outcome was associated with age >65 years (odds ratio [OR] 2.17, 95% confidence interval [CI] 1.20-3.85; p = 0.01), refractory convulsive status epilepticus (OR 2.00, 95% CI 1.04-3.85; p = 0.04), primary brain insult (OR 2.00, 95% CI 1.02-4.00; p = 0.047), and no bystander-witnessed seizure onset (OR 2.49, 95% CI 1.05-5.59; p = 0.04). In our population, systemic secondary brain insults were not associated with outcome in critically ill patients with convulsive status epilepticus. NCT01359332.


Characterization of a temperature-sensitive mouse middle ear epithelial cell line.
Temperature-sensitive mouse middle ear epithelial cells have been successfully established and characterized. Temperature-sensitive middle ear epithelial cell lines are essential for pathophysiologic studies of otitis media. They are useful for studying the pathogen-host interaction, receptor identification, signal transduction, cytokine/mucin production and cellular responses, especially for cell proliferation and differentiation. The purpose of this study was to establish a large T-antigen [simian virus 40 A-gene (SV40)] mutant-immortalized mouse middle ear epithelial cell line for otitis media studies. Primary culture of middle ear epithelial cells was established from the middle ear mucosa of an Immortomouse. The cells were transduced by a temperature-sensitive large T-antigen mutant and cultured for >50 passages. The expression of mRNA transcripts and proteins for epithelial cells was characterized by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry. The temperature-sensitive properties of cells cultured at 33 degree C and 39 degree C were evaluated using 3H-thymidine incorporation, Trypan blue exclusion and peroxisome proliferator-activated receptor-gamma activity. Immortalized middle ear epithelial cells demonstrated a cobblestone-like monolayer culture. The cells expressed mucosal cell markers such as mucins, keratins and collagens. They proliferated at 33 degree C when the SV40 antigen was active and differentiated at 39 degree C when the SV40 antigen was inactive.


Mossy cells in the mouse dentate gyrus: identification in the dorsal hilus and their distribution along the dorsoventral axis.
Previously we showed that large multipolar cells immunoreactive for calretinin and subunits 2 and 3 of amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) type glutamate receptors (GluR2/3) clustered in the ventral hilus of the mouse dentate gyrus and revealed that they were mossy cells. Although such large calretinin immunoreactive cells were not seen in the dorsal hilus, our Golgi study revealed the presence of mossy cells in the dorsal hilus. As we observed large intensely GluR2/3 immunoreactive cells in the dorsal hilus, we suggested that these calretinin negative but intensely GluR2/3 positive large cells in the dorsal hilus were also mossy cells. In the present study we confirmed this identification with several methods. The extracellular tracer labeling studies revealed that all of 47 mossy cells identified morphologically were intensely GluR2/3 positive but calretinin negative, whereas none of 22 non-mossy hilar neurons were intensely GluR2/3 positive. Electron microscopically most of intensely GluR2/3 positive somata and dendritic processes showed the characteristic ultrastructural features of mossy cells. Furthermore, the fimbria-fornix-hippocampal commissure transection procedures induced the calretinin expression in some of these dorsal GluR2/3 immunoreactive cells. On the basis of these observations, we concluded that the vast majority of intensely GluR2/3 immunoreactive large cells in the mouse dorsal hilus were mossy cells. Then we evaluated the presumed difference in the distribution of mossy cells along the dorsoventral axis by the disector. The numerical density of mossy cells was about 1.4 times larger at the ventral level than at the dorsal level, indicating that the dorsoventral difference in the distribution of mossy cells in the mouse hilus was far smaller than that previously speculated.


A device for supplementing breast-feeding.
A device for providing additional nourishment to a breast-fed infant may be constructed to address two key problems of supplemental feeding.


Impact of a Clinical Decision Support System on Inappropriate Prescription of Glucose-lowering Agents for Patients With Renal Insufficiency in an Ambulatory Care Setting.
Inappropriate dosing of glucose-lowering drugs in patients with renal insufficiency can cause severe harm. This study evaluated the short- and long-term effects of clinical decision support systems (CDSS) on inappropriate prescriptions of glucose-lowering agents for patients with renal insufficiency in an ambulatory care setting. This retrospective longitudinal observational study was conducted by using an electronic medical record database and the CDSS log data at Taipei Veterans General Hospital between January 1, 2015, and December 31, 2018. Outpatients who received 7 target glucose-lowering medications and had an estimated glomerular filtration rate <50 mL/min/1.73 m2 were included. Inappropriate prescriptions were defined as a dose, frequency, or daily dose of target drugs that exceeded the dosing recommendations based on renal function. Inappropriate monthly rates were calculated, and the interrupted time series analysis method was used to explore the 1- and 3-year post-implementation effects of CDSS. The major outcome measurements were the level changes and the inappropriate prescription rate trend changes after renal CDSS implementation. The acceptance rates of alerts were also analyzed. A total of 141,037 drug prescriptions were obtained during the study period. In the short-term analysis, the baseline inappropriate rate for overall medications was estimated to range from 30.54% in the first month to 27.06% in month 12. The predicted inappropriate rate 12 months after implementation was 19.35%, corresponding to an estimated 28.49% [(27.06 - 19.35)/27.06] decrease in inappropriate rate. However, after long-term analysis, the predicted inappropriate rate at the end of the study (36 months after implementation) was 18.02%. A total of 27,189 alerts were generated and 628 were accepted during the study period. Thus, after short- and long-term analysis, the overall acceptance rate was 3.06% and 2.31%, respectively. Implementing a CDSS for renal dosing adjustment could significantly decrease the inappropriate prescription rate of glucose-lowing agents among patients with renal insufficiency in an ambulatory setting in the short term, while the long-term effect of a CDSS is limited.


Cytosolic DNA Sensor Upregulation Accompanies DNA Electrotransfer in B16.F10 Melanoma Cells.
In several preclinical tumor models, antitumor effects occur after intratumoral electroporation, also known as electrotransfer, of plasmid DNA devoid of a therapeutic gene. In mouse melanomas, these effects are preceded by significant elevation of several proinflammatory cytokines. These observations implicate the binding and activation of intracellular DNA-specific pattern recognition receptors or DNA sensors in response to DNA electrotransfer. In tumors, IFNβ mRNA and protein levels significantly increased. The mRNAs of several DNA sensors were detected, and DAI, DDX60, and p204 tended to be upregulated. These effects were accompanied with reduced tumor growth and increased tumor necrosis. In B16.F10 cells in culture, IFNβ mRNA and protein levels were significantly upregulated. The mRNAs for several DNA sensors were present in these cells; DNA-dependent activator of interferon regulatory factor (DAI), DEAD (Asp-Glu-Ala-Asp) box polypeptide 60 (DDX60), and p204 were significantly upregulated while DDX60 protein levels were coordinately upregulated. Upregulation of DNA sensors in tumors could be masked by the lower transfection efficiency compared to in vitro or to dilution by other tumor cell types. Mirroring the observation of tumor necrosis, cells underwent a significant DNA concentration-dependent decrease in proliferation and survival. Taken together, these results indicate that DNA electrotransfer may cause the upregulation of several intracellular DNA sensors in B16.F10 cells, inducing effects in vitro and potentially in vivo.


Accidental over-dose of alfathesin under general anaesthesia: case report.
Under general anaesthesia, 35 ml of alfathesin was accidentally injected intravenously. There was no change in the pulse rate and blood pressure. However, two hours later, at the end of the operation, the rectal temperature was 31 degrees C. There were no long-lasting effects of the overdose.


Biomechanical and biological evaluations of novel BPA-free fibre-reinforced composites for biomedical applications.
This aim was to assess the biomechanical and biocompatibility properties of novel glass fibre-reinforced composites (FRCs) with a fluorinated urethane dimethacrylate (FUDMA) resin. Three ratios of FUDMA/TEGDMA (30/70 wt%, 50/50 wt%, 70/30 wt%) and two ratios of control FRCs with bis-GMA/TEGDMA (50/50 wt% and 70/30 wt%) containing long silanized E-glass fibres were prepared. Despite 70 wt% bis-GMA-FRC showed a significantly higher flexural strength (p < 0.05), 50 wt% FUDMA- and bis-GMA-FRCs were not differ from each other. The greatest surface hardness and weight increase after water storage were found in 70 wt% and 30 wt% FUDMA-FRCs, respectively. No significant difference was found in water sorption and solubility among all groups. Average surface roughness was 1.80 ± 0.05 μm, while 70 wt% FUDMA-FRC exhibited the greatest contact angle (p > 0.05). Viabilities and ALP activities of MC3TC-E1 cells in all FUDMA-FRCs were higher than bis-GMA-FRCs after 5 days. To conclude, the novel FUDMA-FRCs are potential substitutes that exhibited superior cytocompatibility properties but comparable biomechanical properties to bis-GMA-FRCs.


Effects of Concurrent Training with Elastic Tubes in Hypertensive Patients: A Blind Controlled Randomized Clinical Trial.
Concurrent training (CT) has been recommended to minimize the deleterious effects of aging. However, few studies have investigated whether this type of training reduces blood pressure in the elderly. Therefore, our objective was to evaluate the effects of CT on the hemodynamic, cardiorespiratory, and muscle strength responses in medicated hypertensive patients. Twenty-three hypertensives (62.65 ± 6.4 years) of both sexes were allocated to the concurrent training group (CTG) or control group (CG). The CTG performed aerobic training (70-85% of reserve heart rate) combined with resistance training with elastic tubes (2sets × 15 repetitions) for 8 weeks. Resting blood pressure (BP), peak oxygen consumption (VO2peak), and right knee and elbow flexion strength were evaluated. A reduction of 6.37% was observed in BP and increases of 16.68% in VO2peak and 16% in muscle strength for right elbow flexion in the CTG compared to CG (p < .05). Intragroup comparisons showed reduction of 5% for BP, and increases of 6.79% for VO2peak, 24.79% for elbow flexion, and 16.47% for knee flexion in the CTG (p < .05), without significant improvement in the CG. CT promoted a reduction in BP, and increased cardiorespiratory fitness and muscular strength of the upper limbs in the hypertensive older adults.


Moonlighting proteins: old proteins learning new tricks.
Recently, several laboratories identifying proteins involved in the complex processes of replication, transcription and tumor suppression found that the 'new' protein they discovered had another, previously identified, function. A single protein with multiple functions might seem surprising, but there are actually many cases of proteins that 'moonlight', or have more than one role in an organism. As well as adding to the number and types of proteins that are known to moonlight, these new examples add to our understanding of the potential importance of moonlighting proteins.


Airline capacity distribution under financial budget and resource consideration.
Capacity distribution is a challenging issue for an airline under financial budget and resource consideration. It is a large-scale optimization problem covering both long-term planning and short-term operating arrangements. This study investigates on the airline capacity distribution problem with financial budget and resource consideration. It contains subproblems of financial budget arrangement, fleet introduction, and fleet assignment. Among them, financial budget is arranged in multiple decision periods, fleet introduction is decided under fixed time points, while fleet assignment is decided under all available time points. To tackle this problem, an integer programming model is formulated for descriptions. Then, an integrated algorithm of modified Variable Neighborhood Search (VNS) and Branch-and-bound (B&B) strategy is developed to find solutions. In detail, a greedy heuristic approach is utilized to generate an initial solution for fleet introduction, the modified B&B strategy is utilized to generate the optimal solution for fleet assignment and the modified VNS is applied to update current solution for a new one with better quality. In addition, budget limit checks are added for financial budget arrangements. Finally, the hybrid algorithm is tested on efficiency and stability. It is also compared to other algorithms which replace the modified VNS by basic VNS, differential evolution and genetic algorithm. Computational results show that performance of our approach is powerful in terms of objective value, convergence speed and stability.


Clonidine modulates the ventrolateral medullary catechol metabolic hyperactivity induced by hypotension.
In vivo electrochemistry allowed recording of a catechol oxidation current in the ventrolateral medulla, caudal to the obex, in anesthetized rats whose ventilatory, metabolic and hemodynamic parameters were rigorously controlled. Hemorrhage or controlled hypotension induced an increase in the metabolism of catecholamines in the A1 noradrenergic group, which remained activated after full hemodynamic recovery. Clonidine (200 micrograms.kg-1 i.p.) given 30 min prior to hemorrhage or immediately before controlled hypotension suppressed partially the increased metabolism of catecholamines especially during the recovery period. This suggests that clonidine preserved phasic reactivity upon circulatory disturbances and decreased tonic hyperactivity following circulatory recovery.


A comparison of patient-controlled analgesia administered by the intravenous or intranasal route during the early postoperative period.
Intranasal administration of lipophilic opioids has been shown to be an effective method of administration which is devoid of major side-effects. Whether it is as effective as intravenous administration for patient-controlled analgesia (PCA) has been investigated for fentanyl and pethidine, but not for diamorphine. This study reports a randomised controlled trial designed to compare the effectiveness of diamorphine administered as PCA utilising either the intranasal or intravenous routes. We investigated 52 consecutive patients undergoing primary lower limb joint replacement surgery. Patients were randomly allocated to receive PCA diamorphine, administered either intravenously (0.5 mg bolus, 3 min lockout) or intranasally (1.0 mg bolus, 3 min lockout). Pain was assessed using a Visual Analogue Score (VAS) at rest and on movement on five occasions over the first 36 h postoperatively. The results demonstrated that patients in the intranasal PCA group had significantly higher VAS scores than the intravenous group, both at rest (intranasal median 35.5 vs. intravenous median 20; p = 0.030) and on movement (intranasal median 64 vs. intravenous median 50; p = 0.016). However, significantly fewer patients in the intranasal group compared with the intravenous group suffered episodes of vomiting (intranasal 0/24 vs. intravenous 6/24 patients; p = 0.022). We suggest that if a maximal reduction in pain score is considered the goal of PCA management, the intravenous route is preferable to the intranasal route.


Glucose Metabolism and Oxygen Availability Govern Reactivation of the Latent Human Retrovirus HTLV-1.
The human retrovirus HTLV-1 causes a hematological malignancy or neuroinflammatory disease in ∼10% of infected individuals. HTLV-1 primarily infects CD4+ T lymphocytes and persists as a provirus integrated in their genome. HTLV-1 appears transcriptionally latent in freshly isolated cells; however, the chronically active anti-HTLV-1 cytotoxic T cell response observed in infected individuals indicates frequent proviral expression in vivo. The kinetics and regulation of HTLV-1 proviral expression in vivo are poorly understood. By using hypoxia, small-molecule hypoxia mimics, and inhibitors of specific metabolic pathways, we show that physiologically relevant levels of hypoxia, as routinely encountered by circulating T cells in the lymphoid organs and bone marrow, significantly enhance HTLV-1 reactivation from latency. Furthermore, culturing naturally infected CD4+ T cells in glucose-free medium or chemical inhibition of glycolysis or the mitochondrial electron transport chain strongly suppresses HTLV-1 plus-strand transcription. We conclude that glucose metabolism and oxygen tension regulate HTLV-1 proviral latency and reactivation in vivo.


It's not who you lose, it's who you are: Identity and symptom trajectory in prolonged grief.
The death of a loved one has been associated with a range of emotional and cognitive impacts, with up to 10% of the bereaved population experiencing a prolonged grief reaction. Direct investigation of the role of self-identity in the maintenance of grief symptoms is limited and has not discriminated between relationship type. This longitudinal study investigated the differences in grief symptoms over time depending on relationship to the deceased person (partner or adult child), as well as the association between long-term grief symptoms and identity, attachment, and cognitive interdependence. Data from bereaved partners and adult children in The Aarhus Bereavement Study at two- and 18-months post-bereavement were included in this study. They completed questionnaires measuring their grief symptoms at both time points, a measure of attachment at Time 1, and measures of the interdependence of their pre- and post-loss identity with the deceased, their cognitive interdependence, and everyday memory retrieval failures at Time 4. Compared with adult children, bereaved partners experienced more intense grief symptoms at both time points. Regression analysis identified that over and above immediate grief symptoms, key predictors of prolonged grief symptoms were a merged post-bereavement identity with the deceased, younger age, and everyday memory retrieval difficulties. Relationship type and pre-bereavement identity contributed to initial but not prolonged grief symptoms. We discuss these findings in terms of the role of interdependence in prolonged grief.


Hypothalamic histamine release by taste stimuli in freely moving rats: possible implication of palatability.
Our previous study indicated that taste information via the chorda tympani (CT) activates the central histaminergic system in anesthetized rats. However, the physiological roles of taste-induced histamine release remain unknown, thus to further investigate the relationship between histamine release and gustatory information, in the present study we investigated the effect of taste stimuli infused intraorally on histamine release using in vivo microdialysis in free moving rats. Consistent with findings from our previous study, application of NaCl and HCl caused significant increases in histamine levels further supporting the suggestion that this phenomenon is attributed to the excitation of the CT. When rats were intraorally infused with quinine HCl (QHCl) solution, a significant increase in hypothalamic histamine release was observed. On the other hand, histamine release was decreased by sucrose and saccharin solutions. When rats were conditioned to acquire taste aversion to sucrose solution or saccharin solution, instead of the histamine decrease seen by the palatable solutions, the pattern of histamine release was similar to that seen by QHCl solution. From these observations, it is concluded that the histamine release by the infusion of these tastants may be explained by two mechanisms-by causing a transient increase after taste stimulation and by causing a decrease relative to the tastant's palatability.


[Silhouettes: electron microscopic photography in bioscience].
The paper describes the first attempts of biological electron microphotography. It starts with a description of the early use of electron microscopy in biology, showing that electron microscopy was used as an extension of former light microscopical studies. Thus, the pictures produced by electron microscopy are interpreted as describing the micro-texture of those structures already seen in light microscopy. That was done irrespective from the specific problems of tissue preparation for electron microscopy. The use of photography in electron microscopy is discussed in more detail. It is shown that in electron microscopy, not the preparation itself which is usually destroyed or damaged during observation in the electron microscope. Thus, biological electron microscopy can be described as a real image science.


Tipping the Scales: Peptide-Dependent Dysregulation of Neural Circuit Dynamics in Alzheimer's Disease.
Identifying effective treatments for Alzheimer's disease (AD) has proven challenging and has instigated a shift in AD research focus toward the earliest disease-initiating cellular mechanisms. A key insight has been an increase in soluble Aβ oligomers in early AD that is causally linked to neuronal and circuit hyperexcitability. However, other accumulating AD-related peptides and proteins, including those derived from the same amyloid precursor protein, such as Aη or sAPPα, and autonomously, such as tau, exhibit surprising opposing effects on circuit dynamics. We propose that the effects of these on neuronal circuits have profound implications for our understanding of disease complexity and heterogeneity and for the development of personalized diagnostic and therapeutic strategies in AD. Here, we highlight important peptide-specific mechanisms of dynamic pathological disequilibrium of cellular and circuit activity in AD and discuss approaches in which these may be further understood, and theoretically and experimentally leveraged, to elucidate AD pathophysiology.


Cereulide-producing strains of Bacillus cereus show diversity.
Producers of cereulide, the emetic toxin of Bacillus cereus, are known to constitute a specific subset within this species. We investigated physiological and genetic properties of 24 strains of B. cereus including two high cereulide producers (600-1,800 ng cereulide mg(-1) wet weight biomass), seven average producers (180-600 ng cereulide mg(-1) wet weight biomass), four low cereulide producers (20-160 ng cereulide mg(-1) wet weight biomass) and 11 non-producers representing isolates from food, food poisoning, human gut and environment. The 13 cereulide producers possessed 16S rRNA gene sequences identical to each other and identical to that of B. anthracis strains Ames, Sterne from GenBank and strain NC 08234-02, but showed diversity in the adk gene (two sequence types), in ribopatterns obtained with EcoRI and PvuII (three types of patterns), in tyrosin decomposition, haemolysis and lecithin hydrolysis (two phenotypes). The cereulide-producing isolates from the human gut represented two ribopatterns of which one was novel to cereulide-producing B. cereus and two phenotypes. We conclude that the cereulide-producing B. cereus are genetically and biochemically more diverse than hitherto thought.


Characterization of a rat model of bortezomib-induced painful neuropathy.
Bortezomib (Velcade®) is a breakthrough treatment for multiple myeloma, significantly improving patient survival. However, its use is limited by painful neuropathy often resulting in dose reduction/cessation of first-line treatment due to lack of treatment. The aim of this study was to characterize a clinically relevant rat model of bortezomib-induced painful neuropathy, using established evoked measures and novel ethological techniques, to aid drug discovery. Adult male Sprague-Dawley rats were injected i.p. with 0.1 and 0.2 mg·kg-1 bortezomib, or its vehicle, on days 0, 3, 7 and 10. Multiple behavioural approaches were utilized: mechanical hypersensitivity, cold allodynia, heat hypersensitivity, motor co-ordination, burrowing and voluntary wheel running. At maximal bortezomib-induced mechanical hypersensitivity, 200 mg·kg-1 ethosuximide/vehicle and 100 mg·kg-1 phenyl N-tert-butylnitrone (PBN)/vehicle were administered i.p. in separate experiments, and mechanical hypersensitivity assessed 1, 3 and 24 h later. Bortezomib induced dose-related mechanical hypersensitivity for up to 80 days. Bortezomib induced short-term cold allodynia, but no significant change in heat hypersensitivity, motor co-ordination, voluntary wheel running and burrowing behaviour compared to vehicle-treated controls. Systemic PBN and ethosuximide significantly ameliorated bortezomib-induced mechanical hypersensitivity. These data characterize a reproducible rat model of clinical-grade bortezomib-induced neuropathy demonstrating long-lasting pain behaviours to evoked stimuli. Inhibition by ethosuximide and PBN suggests involvement of calcium and/or ROS in bortezomib-induced painful neuropathy. These drugs could be used as preclinical positive controls to assess novel analgesics. As ethosuximide is widely used clinically, translation to the clinic to treat bortezomib-induced painful neuropathy may be possible.


Ginsenoside content and variation among and within American ginseng (Panax quinquefolius L.) populations.
The contents of five ginsenosides (Rg1, Re, Rb1, Rc and Rd) were measured in American ginseng roots collected from 10 populations grown in Maryland. Ginsenoside contents and compositions varied significantly among populations and protopanaxatriol (Rg1 and Re) ginsenosides were inversely correlated within root samples and among populations. The most abundant ginsenoside within a root and by population was either Rg1 or Re, followed by Rb1. Ginseng populations surveyed grouped into two chemotypes based on the relative compositions of Rg1 and Re. Four populations, including the control population in which plants were grown from TN and WI seed sources, contained roots with the recognized chemotype for American ginseng of low Rg1 composition relative to Re. The remaining 6 populations possessed roots with a distinctive chemotype of high relative Rg1 to Re compositions. Chemotype did not vary by production type (wild versus cultivated) and roots within a population rarely exhibited chemotypes different from the overall population chemotype. These results provide support for recent evidence that relative Rg1 to Re ginsenoside contents in American ginseng roots vary by region and that these differences are likely influenced more by genotype than environmental factors. Because the physiological and medicinal effects of different ginsenosides differ and can even be oppositional, our findings indicate the need for fingerprinting ginseng samples for regulation and recommended usage. Also, the High Rg1/Low Re chemotype discovered in MD could potentially be used therapeutically for coronary health based on recent evidence of the positive effects of Rg1 on vascular growth.


Safety and efficacy of herbal extracts to restore respiratory health and improve innate immunity in COVID-19 positive patients with mild to moderate severity: A structured summary of a study protocol for a randomised controlled trial.
Primary Objective • To assess the efficacy of herbal extracts in boosting innate immunity of patients with COVID-19 infection. Secondary Objectives • To assess the efficacy of herbal extracts in restoring respiratory health • To assess the efficacy of Cap. IP in early recovery of patients and decline in viral load • To assess the safety of herbal extracts TRIAL DESIGN: This is a single centre, randomized, 2-arm, parallel group, double blind, 1:1 ratio, controlled, exploratory trial with a study period of 30 days from the day of enrolment. Patients attending the COVID treatment centre at Yashwantrao Chavan Memorial Hospital, Nehrunagar, Pimpri, Pune, India were screened for their participation in the study. Patients who were known COVID-19 positive (with positive RT-PCR), eligible and willing were enrolled in the study. The intervention in the trial has a background in 'Ayurved'. Intervention Arm: Two capsules, Investigational Product (IP) - 1 - 400mg and Investigational Product - 2 - 450mg, containing herbal extracts (a blend of water and CO2 extracts) of Shunthi (Zingiber officinale (Ginger), Vidanga (Embelia ribes), Yashtimadhu (Glycyrrhiza glabra), Haritaki (Terminalia chebula), Guduchi (Tinospora cordifolia), Shatavari (Asparagus racemosus), Aamalaki (Emblica officinalis), Pippali (Piper longum) and calcined Zinc, Shankha bhasma. Placebo Arm: Edible starch ~ 450 mg. The look and feel of IP and of Placebo boxes were very similar. Patients are to take two capsules (one each of IP-1 and IP-2) twice a day for 15 days, and from the 16th day, one capsule of IP-2 twice a day up-to day 30. Capsules are to be administered orally with plain water. The IP is to be taken with all other concomitant medicines prescribed by the treating physician/doctor. The dose of each component in the IP is very safe to administer. The investigational products are registered products with the Indian Government and have been used for more than 6 months in various health conditions but not for COVID-19. Primary Outcome: Efficacy of the herbal extracts in COVID 19 positive patients (in declining viral load: time-point: 4 days and early recovery) Secondary Outcomes: Efficacy of the herbal extracts as an immune-modulator - TH1, TH2, Th17, IL6, NK Cells and CD markers; Immunoglobulin IGG (Serum); Immunoglobulin IGM (Serum) - at 30 days. Efficacy of the investigational product in reducing sequela of the disease Safety analysis (Liver Function Test and Kidney Function Test) including serious allergic reaction of: rash, itching/swelling, severe dizziness, trouble breathing. An alphanumeric coded set of IP/Placebo containers will be used. Participants will be automatically randomized to two groups in the ratio 1:1. Participants, caregivers and investigators were blinded. A total of more than 60 and up to 75 patients were to be enrolled in the study into the two groups, considering drop-outs. 72 were enrolled with 37 into the intervention group and 35 into the placebo group. Protocol number: CoviQuest-01 Protocol version number: 1.2 Protocol Date: 1st July 2020 The recruitment period is completed for the trial. Date of 1st patient enrolment was 11th Aug 2020 and the last patient was enrolled on 3rd of September 2020. This is to state that it was a late submission from authors for publication of the protocol to the BMC, after enrolment in the study was over. Last Participant's last follow-up is scheduled on 5th October 2020 TRIAL REGISTRATION: The trial was prospectively registered with the CTRI (Clinical Trial Registry of India). Registration number is CTRI/2020/07/026570 . Registered on 14 July 2020 FULL PROTOCOL: The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). In the interest in expediting dissemination of this material, the familiar formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol.


Functional interplay between histone H1 and HMG proteins in chromatin.
The dynamic interaction of nucleosome binding proteins with their chromatin targets is an important element in regulating the structure and function of chromatin. Histone H1 variants and High Mobility Group (HMG) proteins are ubiquitously expressed in all vertebrate cells, bind dynamically to chromatin, and are known to affect chromatin condensation and the ability of regulatory factors to access their genomic binding sites. Here, we review the studies that focus on the interactions between H1 and HMGs and highlight the functional consequences of the interplay between these architectural chromatin binding proteins. H1 and HMG proteins are mobile molecules that bind to nucleosomes as members of a dynamic protein network. All HMGs compete with H1 for chromatin binding sites, in a dose dependent fashion, but each HMG family has specific effects on the interaction of H1 with chromatin. The interplay between H1 and HMGs affects chromatin organization and plays a role in epigenetic regulation.


Footprinting SHAPE-eCLIP Reveals Transcriptome-wide Hydrogen Bonds at RNA-Protein Interfaces.
Discovering the interaction mechanism and location of RNA-binding proteins (RBPs) on RNA is critical for understanding gene expression regulation. Here, we apply selective 2'-hydroxyl acylation analyzed by primer extension (SHAPE) on in vivo transcripts compared to protein-absent transcripts in four human cell lines to identify transcriptome-wide footprints (fSHAPE) on RNA. Structural analyses indicate that fSHAPE precisely detects nucleobases that hydrogen bond with protein. We demonstrate that fSHAPE patterns predict binding sites of known RBPs, such as iron response elements in both known loci and previously unknown loci in CDC34, SLC2A4RG, COASY, and H19. Furthermore, by integrating SHAPE and fSHAPE with crosslinking and immunoprecipitation (eCLIP) of desired RBPs, we interrogate specific RNA-protein complexes, such as histone stem-loop elements and their nucleotides that hydrogen bond with stem-loop-binding proteins. Together, these technologies greatly expand our ability to study and understand specific cellular RNA interactions in RNA-protein complexes.


Characterization of freezability-associated metabolites in boar semen.
Sperm cryopreservation maintains the diversities of porcine genetic resources and improves utilization efficiency of boar semen in artificial insemination practices. Freezability of boar semen presents remarkable differences among individuals. However, metabolic markers for boar semen freezability in both sperm and seminal plasma largely remain unknown. The present study thus aims to determine differences in metabolites of sperm and seminal plasma between poor (PF) and good (GF) freezability semen from a Chinese native pig and screen potential markers for semen freezability. A total of 72,048 metabolites in sperm and 66,551 metabolites in seminal plasma were identified by liquid chromatography-mass spectrometry, respectively. The proportion of lipid molecules among all metabolites in both sperm and seminal plasma was the maximum regardless of negative or positive mode. Furthermore, we identified 21 differentially expressed metabolites (DEMs) in sperm and 185 DEMs in seminal plasma between PF and GF group. Additionally, clustering analysis showed that DEMs in sperm and seminal plasma exhibited significant changes between PF and GF group. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that DEMs in sperm were mainly enriched in metabolic pathways of amino acids and caffeine. DEMs in seminal plasma were associated with AMPK and cAMP signaling pathways. Taken together, these results demonstrate that sperm and seminal plasma of native pigs present differential metabolome between PF and GF semen.


Polymorphisms in CARD8 and NLRP3 are associated with extrapulmonary TB and poor clinical outcome in active TB in Ethiopia.
Innate immunity is a first line defense against Mycobacterium tuberculosis infection where inflammasome activation and secretion of the pro-inflammatory cytokine IL-1beta, plays a major role. Thus, genetic polymorphisms in innate immunity-related genes such as CARD8 and NLRP3 may contribute to the understanding of why most exposed individuals do not develop infection. Our aim was to investigate the association between polymorphisms in CARD8 and NLRP3 and active tuberculosis (TB) as well as their relationship to treatment outcome in a high-endemic setting for TB. Polymorphisms in CARD8 (C10X) and NLRP3 (Q705K) were analysed in 1190 TB patients and 1990 healthy donors (HD). There was a significant association between homozygotes in the CARD8 polymorphism and extrapulmonary TB (EPTB), which was not the case for pulmonary TB or HDs. Among TB-patients, there was an association between poor treatment outcome and the NLRP3 (Q705K) polymorphism. Our study shows that inflammasome polymorphisms are associated with EPTB and poor clinical outcome in active TB in Ethiopia. The practical implications and determining causal relationships on a mechanistic level needs further study.


Technical Aspects Related to the Analysis of Circulating microRNAs.
Specific and sensitive noninvasive biofluid-based biomarkers are always needed in the laboratory diagnosis of diseases. Biomarkers are applied not only for diagnostic purposes but for stratifying a disease and for assessing the therapy response or disease progression. MicroRNAs (miRNAs) are short noncoding RNA molecules regulating gene expression posttranscriptionally. They are frequently dysregulated in many physiological and pathophysiological conditions. miRNAs are present in the circulation and in other biofluids that are common matrices for clinical laboratory testing that has raised the possibility that miRNAs may serve as novel biomarkers. Their excellent stability also supports the possibility that miRNAs once will be routinely used biomarkers in clinical practice. From an analytical point of view, there are many factors (starting material, sample storage and processing, different RNA extraction and detection methods, intra- and interassay variability, and assay interferences) to consider if a miRNA as biomarker is aimed to be introduced as a clinical laboratory test. Despite several pre-analytical and analytical factors that still need standardization, a significant number of studies have been published about the potential role of circulating miRNAs as biomarkers. Due to the lack of standardization of methods, there are a lot of discrepancies among results. In this chapter, we aimed to summarize the current findings about circulating miRNAs focusing on the analytical points related to miRNAs measurements from biofluids.


C4 uremic variant: an acquired C4 allotype.
The major histocompatibility complex (MHC)-linked complotype region includes alleles for B, C2, and C4 loci. These loci are closely linked to each other and to HLA-DR on chromosome 6. The duplicated C4 loci, C4A and B, are especially polymorphic. In seven patients with renal insufficiency, we observed a C4 variant with electrophoretic mobility between C4B2 and C4B3. Four of these patients were detected during a study of MHC markers in membranoproliferative glomerulonephritis. Complete complotype and HLA data from families of five of the seven patients demonstrated that the variant was not inherited. The pattern was revealed by immunofixation electrophoresis and also by C4-specific hemolytic overlay. In serial plasma specimens taken from one patient, the C4 variant appeared only after the patient became uremic. However, the variant could not be produced in normal plasma after incubation with C4-depleted uremic plasma. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions of immunoprecipitated C4 from these patients showed C4A and C4B alpha chains of normal molecular mass; incompletely processed forms of C4 were not observed. We believe that this variant is probably acquired in the presence of uremia and may represent the C4B2.9 allele found by Wank and co-workers in many patients with glomerulonephritis. Family studies are mandatory to distinguish genetic variants from acquired alterations in the C4 phenotype.


Hypnosis meets neuropsychology: simulating visuospatial neglect in healthy participants.
Neglect patients are not aware of stimuli in the contralesional space. We aimed to simulate neglect-like behaviour in healthy participants, by asking them to orient their visuospatial attention in two conditions: non-hypnotic suggestion and post-hypnotic suggestion. Results showed that directing visuospatial attention to one side of space caused neglect of stimuli in the opposite side of space, but only when participants were under post-hypnotic suggestion. Furthermore, directing visuospatial attention to the right side of space caused more neglect of left-sided stimuli than directing visuospatial attention to the left side of space did for right-sided stimuli. We propose that post-hypnotic suggestion can be a useful tool for (de)activating neurocognitive mechanisms underlying visuospatial awareness, a function that is fundamental for our survival. The use of post-hypnotic suggestion could be applied to the study of many domains of cognitive neurosciences (e.g., neurocognitive rehabilitation).


Binding properties of oligonucleotides containing a modified 2'-deoxyuridine with a thymine ended linker to pair with 2'-deoxyadenosine.
Oligonucleotides containing in the place of thymidine the nucleoside 2, a 2'-deoxyuridine harbouring at C-5 a thymine ended linker, were found to undergo base pairing with the opposite 2'-deoxyadenosine. However, the corresponding duplexes are significantly destabilised as compared to the fully natural ones.


A rational approach to assess volume status in patients with decompensated heart failure.
Although the past few decades have yielded significant improvements in the management of cardiomyopathy, heart failure remains a leading cause of hospitalization as well as readmissions, both in the United States and worldwide. Accurate assessment of volume status in these patients is of great importance because it guides initial therapies and aids in determining suitability for discharge. However, this assessment can prove to be challenging and, therefore, must be based on multiple data points and techniques. This review examines the various modalities of volume assessment in the patient with decompensated heart failure.


Therapeutic potential of low-dose IL-2 in a chronic GVHD patient by in vivo expansion of regulatory T cells.
Chronic graft-versus-host disease (cGVHD) is a common complication following allogeneic hematopoietic stem cell transplantation (HSCT), which is characterized by autoimmune like inflammatory responses and reduced levels of regulatory T cells (Tregs). Recently, the use of low-dose IL-2 has been reported to selectively increase Tregs and therefore facilitate immune regulation and promote clinical improvements in cGVHD patients. In this report, we describe the case of a cGVHD patient who was treated with daily low-dose IL-2 therapy. Our observations demonstrate that low-dose IL-2 could induce significant expansion of Tregs in vivo leading to improved Treg/Th17 ratios. The patient showed moderate clinical benefits suggesting that multiple factors may be involved in the immunological responses. Therefore, while the therapeutic potential of low-dose IL-2 is promising, strategic approaches may be needed to induce a clinically significant and sustained Treg effect.


Candida konsanensis sp. nov., a new yeast species isolated from Jasminum adenophyllum in Thailand with potentially carboxymethyl cellulase-producing capability.
A new yeast species (KKU-FW10) belonging to the Candida genus was isolated from Jasminum adenophyllum in the Plant Genetic Conservation Project under The Royal Initiative of Her Royal Highness Princess Maha Chakri Sirindhorn area, Chulabhorn Dam, Konsan district within Chaiyaphum province in Thailand. The strain was identified via analysis of nucleotide sequences from the D1/D2 domain of 26S ribosomal DNA and based on its morphological, physiological and biochemical characteristics. The sequence obtained from yeast isolate KKU-FW10 was 97 percent identical to that of Candida chanthaburiensis (GenBank accession number AB500861.1), with 506/517 (nucleotides identity/total nucleotides) matching nucleotides, nine substitutions and two gaps being detected. This species belonged to the Candida clade. Regarding morphological characteristics, isolate KKU-FW10 presents cream-colored butyrous colonies, vegetative reproduction through budding and, round cells without filaments or ascospores. The major ubiquinone detected was Q-9. The above results suggest that isolate KKU-FW10 is a new member of the genus Candida, and the name Candida konsanensis is proposed for this yeast. The type strain of the new species is KKU-FW10(T) (= BCC 52588(T), = NBRC 109082(T), = CBS 12666(T)). In addition, this KKU-FW10 could potentially produce 58.24 Units/ml of carboxymethyl cellulase when it was cultured in YP broth containing 1.0 % carboxymethyl cellulose for 24 h.


Association of preoperative conization with recurrences after laparoscopic radical hysterectomy for FIGO 2018 stage IB1 cervical cancer.
To evaluate association of preoperative conization with recurrences after laparoscopic radical hysterectomy (LRH) for FIGO 2018 stage IB1 cervical cancer. This is a retrospective single-center study. Patients who underwent LRH for cervical cancer with squamous, adenosquamous and adenocarcinoma subtype from January 2014 to December 2018 were reviewed. All patients were restaged according to the 2018 FIGO staging system. Those who were in FIGO 2018 stage IB1 met the inclusion criteria. General characteristics and oncologic outcomes including recurrence-free survival (RFS) were analyzed. A total of 1273 patients were included in the analysis. 616 (48.4%) patients underwent preoperative biopsy, and 657 (51.6%) patients underwent conization. Residual disease was observed in 822 (64.6%) patients. During a median follow-up of 50.30 months, 30 (2.4%) patients experienced recurrence. The univariate analysis showed that patients who had larger tumor diameter, the presence of residual tumor at final pathology, and underwent adjuvant treatment had a significant higher risk of recurrence (P < 0.01). Conversely, patients who underwent conization were significantly less likely to experience recurrence (P = 0.001). In the multivariate analysis, the independent risk factor associated with an increased risk of recurrence was resident macroscopic tumor (HR: 38.4, 95% CI 4.20-351.64, P = 0.001). On the contrary, preoperative conization was associated with a significantly lower risk of recurrence (HR: 0.26; 95% CI 0.10-0.63, P = 0.003). The Kaplan-Meier curves showed patients who underwent conization had improved survival over those who underwent biopsy (5 year RFS: 98.6 vs 95.1%, P = 0.001). The 5 year RFS of patients with residual tumor was significantly different (R0: 99.2%, R1: 97.4%, R2: 93.6%, P < 0.001), especially the patients with residual macroscopic tumor after conization (R0: 99.5%, R1: 99.0%, R2:92.4%, P = 0.006). Preoperative conization and the absence of residual tumor at the time of surgery might play a protective role in patients with FIGO 2018 IB1 cervical cancer following LRH, which support the theory of the influence of intraoperative tumor spread during radical hysterectomy. Further prospective evidence is needed.


COVID-19: self-reported reductions in physical activity and increases in sedentary behaviour during the first national lockdown in the United Kingdom.
The United Kingdom (UK) government imposed its first national lockdown in response to COVID-19 on the 23rd of March 2020. Physical activity and sedentary behaviour levels are likely to have changed during this period. An online survey was completed by n = 266 adults living within the UK. Differences in day-to-day and recreational physical activity (at moderate and vigorous intensities), travel via foot/cycle, and sedentary behaviour were compared before and during the initial COVID-19 lockdown. The median level of total weekly physical activity significantly reduced (- 15%, p < 0.001) and daily sedentary time significantly increased (+ 33%, p < 0.001). The former was caused by a significant reduction in weekly day-to-day physical activity at moderate intensities (p < 0.001), recreational activities at vigorous (p = 0.016) and moderate (p = 0.030) intensities, and travel by foot/cycle (p = 0.031). Sub-group analyses revealed that some populations became disproportionally more physically inactive and/or sedentary than others, such as those that were: living in a city (versus village), single (versus a relationship), an athlete (versus non-athlete), or earning an average household income < £25,000 (versus > £25,000). Now that the UK is transitioning to a state of normal living, strategies that can help individuals gradually return to physical activities, in accordance with the 2020 WHO guidelines, are of paramount importance to reducing risks to health associated with physical inactivity and sedentary behaviour.


Rumination and behavioural factors in Parkinson's disease depression.
Parkinson's disease is associated with high rates of depression. There is growing interest in non-pharmacological management including psychological approaches such as Cognitive Behaviour Therapy. To date, little research has investigated whether processes that underpin cognitive models of depression, on which such treatment is based, apply in patients with Parkinson's disease. The study aimed to investigate the contribution of core psychological factors to the presence and degree of depressive symptoms. 104 participants completed questionnaires measuring mood, motor disability and core psychological variables, including maladaptive assumptions, rumination, cognitive-behavioural avoidance, illness representations and cognitive-behavioural responses to symptoms. Regression analyses revealed that a small number of psychological factors accounted for the majority of depression variance, over and above that explained by overall disability. Participants reporting high levels of rumination, avoidance and symptom focusing experienced more severe depressive symptoms. In contrast, pervasive negative dysfunctional beliefs did not independently contribute to depression variance. Specific cognitive (rumination and symptom focusing) and behavioural (avoidance) processes may be key psychological markers of depression in Parkinson's disease and therefore offer important targets for tailored psychological interventions.


Chromosomal aberration in peripheral lymphocytes and doses to the active bone marrow in radiotherapy of prostate cancer.
Radiotherapy plays an important role in the management of prostate cancer. Epidemiological data indicate a small but significant risk of radiation-induced leukemia after radiotherapy which might be related to the high mean bone marrow dose associated with radiotherapy of prostate cancer. The purpose of the study was to investigate the relation between the mean bone marrow dose and unstable chromosome aberrations in peripheral blood lymphocytes in patients undergoing conformal radiotherapy for prostate cancer as a possible indicator of risk. Endometrial cancer patients were also included for comparison. Nine patients, six with prostate cancer (60-73 years old) and three with endometrial cancer (61-81 years old) treated with radiotherapy were included in the study. The non-bony spaces inside the pelvic bones were outlined on every CT slice using the treatment planning system and mean doses to the bone marrow calculated. Blood samples of the patients were obtained at different times before, during and at the end of treatment. Lymphocytes were cultured in the usual way and metaphases scored for dicentric aberrations. 46 samples from nine patients were obtained. The mean number of metaphases analyzed per sample was 180 with a range from 52 to 435. The mean bone marrow doses for prostate cancer patients ranged from 2.8 to 4.2 Gy and for endometrial cancer patients from 12.8 to 14.8 Gy. The aberration yield increased with the planning target volume and the mean bone marrow dose. The yield of dicentric aberrations for prostate cancer patients correlated closely with the mean bone marrow dose albeit the induction of dicentrics occurred in mature T lymphocytes most of which were probably in transit through the irradiated volumes. Therefore, the observed relationship between dicentrics and mean bone marrow doses are indirect.


Value of stereotactic 11-gauge vacuum-assisted breast biopsy in non-palpable suspicious calcifications: an eight-year single institution experience with 587 patients.
Vacuum-assisted breast biopsy (VABB) has been routinely recommended for stereotactic intervention in cases of isolate mammographically-detected calcifications. Herein we aimed to evaluate and compare the diagnostic consistency and accuracy of calcified and noncalcified specimens obtained from same sites of sampling on mammography-visible calcifications. In addition, we presented the biopsy procedure and retrospectively evaluated the usefulness of VABB as well as the complications of this technique over an eight-year experience in our centre. This single-institution observational cohort study included 587 patients referred for stereotactic 11-gauge VABB of 594 mammographically-detected calcifications between January 2010 and December 2018. The rate of histopathological underestimation, the false negative, the diagnostic consistency and accuracy between calcified and noncalcified specimens of VABB were comprehensively evaluated based on the surveillance data and final histopathological result of the surgical specimens. In total, 594 biopsy procedures were performed in 587 patients (mean age 46 years, range, 21-80 years). The average number of biopsy specimens was 14.7 (range, 9-21) per lesion. VABB pathological results revealed 471 (79.3%) benign, 39 (6.6%) high-risk, and 84 (14.1%) malignant cases. The diagnostic inconsistency between calcified and noncalcified specimens was 14.6% (105/123) for high-risk and malignant lesions. Furthermore, calcified specimens exhibited higher diagnostic accuracy of malignant lesion as compared with the noncalcified specimens (97.7% versus 82.6%, respectively). Underestimation rate for high-risk lesions and in situ carcinoma was 5.1% and 54.1%, respectively, along with a false negative rate of 6.25%. In addition, mild complications were reported with high patient tolerance. Stereotactic 11G-VABB might be preferred for the investigation of non-palpable mammographically-detected calcifications in terms of accuracy and safety profile. The high prevalence of diagnostic discordance between the specimens with and without calcifications revealed a higher value of calcified specimens in diagnosing high-risk and malignant calcifications.


Membrane intercalation-enhanced photodynamic inactivation of bacteria by a metallacycle and TAT-decorated virus coat protein.
Antibiotic resistance has become one of the major threats to global health. Photodynamic inactivation (PDI) develops little antibiotic resistance; thus, it becomes a promising strategy in the control of bacterial infection. During a PDI process, light-induced reactive oxygen species (ROS) damage the membrane components, leading to the membrane rupture and bacteria death. Due to the short half-life and reaction radius of ROS, achieving the cell-membrane intercalation of photosensitizers is a key challenge for PDI of bacteria. In this work, a tetraphenylethylene-based discrete organoplatinum(II) metallacycle (1) acts as a photosensitizer with aggregation-induced emission. It self-assembles with a transacting activator of transduction (TAT) peptide-decorated virus coat protein (2) through electrostatic interactions. This assembly (3) exhibits both ROS generation and strong membrane-intercalating ability, resulting in significantly enhanced PDI efficiency against bacteria. By intercalating in the bacterial cell membrane or entering the bacteria, assembly 3 decreases the survival rate of gram-negative Escherichia coli to nearly zero and that of gram-positive Staphylococcus aureus to ∼30% upon light irradiation. This study has wide implications from the generation of multifunctional nanomaterials to the control of bacterial infection, especially for gram-negative bacteria.


Usefulness of the four-variable formula on serial electrocardiograms in detecting subtle anterior myocardial infarction.
To perform serial electrocardiogram (ECG) analyses in patients with subtle ECG changes in the anterior leads and evaluate the performance of the four-variable formula in detecting left anterior descending (LAD) coronary artery occlusion. This prospective study included patients admitted to the emergency department with acute chest pain between April 2021 and January 2023, whose initial ECG was not diagnostic but indicated suspicion of myocardial infarction (MI) and who underwent percutaneous coronary intervention in their follow-up. The control group consisted of patients who were diagnosed with benign variant ST-segment elevation (BV-STE) due to ST-segment elevation (STE) of at least 1 mm in the anterior lead, who had normal cardiac troponin levels, and who presented with non-cardiac chest pain. Following admission, six ECGs were taken at 10-min intervals. The scores of all patients were calculated with the four-variable formula on serial ECGs and compared between the groups. A total of 232 patients, including 116 with anterior MI and 116 with BV-STE, were included in the study. When the cut-off value for the four-variable formula was taken as ≥18.2, the sensitivity, specificity, and diagnostic accuracy of the first ECG were determined to be 82.7%, 85.3%, and 83.6%, respectively. We found that the four-variable formula had the highest sensitivity, specificity, and diagnostic accuracy in detecting LAD occlusion for the ECG taken at the 20th minute (83.6%, 89.6%, and 86.2%, respectively). The four-variable formula was found to be a valid method for the differentiation of STEMI and BV-STE in patients with subtle ECG changes. While managing this patient group, using serial ECGs rather than a single ECG to evaluate the clinical status of patients can help clinicians make more accurate decisions.


Bifunctional derivative of p,p'-dichlorochalcone. Part III. Synthesis and study for cytotoxic activity of a new compound, 2-[2,2-bis(4-chlorophenyl)ethyl]-2-(4-chlorophenyl)-thiazolidin-4-one from p,p'-dichlorochalcone.
The synthesis of 2-(2,2-bis(4-chlorophenyl)ethyl]-2-(4-chlorophenyl)-thiazolidin-4-one (3) from p,p'-dichlorochalcone (1) via 1,3,3-tris(4-chlorophenyl)-propan-1-one (2) using thioglycollic acid in the presence of ammonium carbonate is described. Structural assignment, stereochemistry and biological assay are discussed.


P53 mutations in tissue from Danish ovarian cancer patients: from the Danish "MALOVA" ovarian cancer study.
The p53 gene, a tumor suppressor gene located on the short arm of chromosome 17 (17p13), has been found mutated in 30-80% of epithelial ovarian cancers (OC), with the most frequently detected mutations in the conserved regions of the gene. A small number of studies investigated the survival of patients with p53 mutations in OC, but their conclusions are not in agreement. We analyzed the frequency of p53 mutations in 124 Danish women with OC, using Single-Stranded Conformation Polymorphism analysis in addition with DNA sequencing and evaluated if mutations correlated with clinicopathological parameters and with patient survival. Thirty-five (28%) ovarian tumors were found to contain one or more p53 variations, two of which were considered polymorphisms. Twenty-seven (82%) mutations were single nucleotide substitutions of which 23 (85%) were missense mutations and therefore led to amino acid substitutions. Significantly shorter survival was found for stage III/IV patients with a p53 missense mutation compared to stage III/IV OC patients with wild type p53 (P = 0.0018). Multivariate Cox regression analysis restricted to 107 OC patients with a p53 missense mutation or p53 wild type in the tumor tissue and with information on radicality of primary surgery showed that missense p53 mutation (HR = 2.5, 95% CI: 1.21-4.98), radicality after primary surgery (HR = 1.7, 95% CI: 1.04-2.88), tetranectin (mg/l: HR = 0.78, 95% CI: 0.67-0.91) and stage (I vs. III: HR = 0.30, 95% CI: 0.10-0.92, II vs. III: HR = 0.24, 95% CI: 0.05-1.05, IV vs. III: HR = 2.70, 95% CI: 1.22-5.98) were independent prognostic factors. Missense mutations in the conserved regions of p53 may be of prognostic value in Danish OC patients.


Favorable colorectal cancer mortality-to-incidence ratios in countries with high expenditures on health and development index: A study based on GLOBOCAN database.
Global variation in the incidence and outcomes of colorectal cancer (CRC) is associated with many factors, among which screening policies and early treatment play substantial roles. However, screening programs and intense treatment are expensive and require good health care systems. For CRC, no clear association has yet been established between clinical outcomes and health care disparities. We used the mortality-to-incidence ratio (MIR) of CRC as a measure of clinical outcomes for comparison with the Human Development Index (HDI), current health expenditure (CHE), and current health expenditure as a percentage of gross domestic product (CHE/GDP) using linear regression analyses. We included 171 countries based on data from the GLOBOCAN 2018 database. We found that the regions with the lowest MIRs for CRC are Oceania and North America. A significant correlation was observed between incidence, mortality and HDI, CHE, and CHE/GDP among the countries enrolled. Furthermore, lower MIRs of CRC significantly correlated with higher HDI, CHE, and CHE/GDP (P < 0.001, P < 0.001, and P < 0.001, respectively). : CRC MIRs tend to be most favorable in countries with high health care expenditures and a high HDI.


A new model system for investigation of ionic channels in filamentous fungi: evidence for existence of two K+-permeable ionic channels in Phycomyces blakesleeanus.
A technique was developed to obtain viable cytoplasmic droplets, enabling the formation of a gigaohm seal. Such cytoplasmic droplets can be used for characterization of plasma membrane ion channels from filamentous fungi by patch-clamp technique. Two K(+) ionic channels are characterized with a conductance of 43 pS and 74 pS.


Regulation of connexons composed of human connexin26 (hCx26) by temperature.
This report shows that temperature is a latent regulator of the voltage-dependent conductance of hemichannels composed of hCx26. The latter were expressed in Xenopus oocytes by injection of a mixture of hCx26 cRNA and antisense of endogenous Cx38 (anti-Cx38). At 24-25 degrees C, voltage clamp of oocytes at potentials above -40 mV evoked outward currents which were not observed in control oocytes. These currents were reversibly affected by change in temperature. Increasing temperature of the bath solution amplified gradually, whereas decreasing bath temperatures below 20 degrees C reduced the current. Furthermore analysis revealed that temperature-dependent increase of the conductance of the hemichannels did not correlate with a change of the apparent gating charge, whereas the half-activation voltage V(1/2) of the hemichannel was affected by a temperature change. It is proposed that this finding correlates with a temperature-dependent transition into an open state above 20 degrees C. In addition, a temperature-dependent release of Lucifer Yellow from loaded liposomes containing reconstituted purified and hCx26 hemichannels was observed, which indicate that a temperature-dependent regulation of the permeability of hCx26 hemichannels is not related to intracellular mediators. The involvement of temperature to modulate hemichannels as well as of the corresponding gap junction channel composed of hCx26 at physiological condition is discussed.


The role of miRNAs in the pheochromocytomas.
Pheochromocytoma (PCC) is a catecholamine-producing and neuroendocrine tumor, which originates from chromaffin cells in adrenal medulla or at extra-adrenal sites. The clinical symptoms of PCC patients were including paroxysmal hypertension and cardiovascular crises. Despite the identification of many molecular-level basics involved in these pathogenesis processes, the 5-year survival of advanced stage PCC is <40 %. The 5-year survival of advanced stage PCC is <40 %. microRNAs (miRNAs) are a class of single-strand, highly conserved, small, non-coding RNAs which inhibit gene expression by binding to the target messenger RNA (mRNA) 3'UTR to induce translational repression or their degradation. Increasing data have shown that the deregulated expression of miRNAs has been implicated in the diagnosis and treatment of tumors including PCC. In this review, we summarized the expressions and roles of miRNAs in PCC and comment on the potential role of miRNAs in improving diagnosis, prognosis, and treatment of PCC.


Spatial variation of metal bioaccumulation in the hydrothermal vent mussel Bathymodiolus azoricus.
The variability of the bioaccumulation of metals (Ag, Cd, Cu, Fe, Mn and Zn) was extensively studied in the mussel Bathymodiolus azoricus from five hydrothermal vent sites inside three main vent fields of increasing depth along the Mid-Atlantic Ridge: Menez Gwen, Lucky Strike and Rainbow. Metal bioaccumulation varied greatly between vent fields and even between sites inside a vent field with B. azoricus showing a great capacity to accumulate metals. The bioaccumulation of these metals also varied significantly among tissues. The main target was the gills where metals were mainly associated with soluble compounds whereas in the digestive gland they were mainly associated with insoluble compounds. Storage of metals under insoluble forms in B. azoricus seems to be a major pathway for the detoxification of both essential and non-essential metals. Mussels from the studied fields can be discriminated following their metallic load but the segregation relies partially on the composition of the metal-enriched fluids.


Correction: Resistance profile of the HIV-1 maturation inhibitor GSK3532795 in vitro and in a clinical study.
[This corrects the article DOI: 10.1371/journal.pone.0224076.].


A Rare Presentation of a Non-Asian Female With Metastatic Non-small-cell Lung Cancer Harboring EGFR L747P Mutation With Clinical Response to Multi-targeted Epigenetic and EGFR Inhibition.
Activating mutations of the epidermal growth factor receptor (EGFR) gene have been utilized to predict the effectiveness of EGFR tyrosine kinase inhibitor (TKI) therapy. The most common EGFR mutations are exon 19 deletion and exon 21-point mutation, which are sensitive to EGFR TKI. However, rare/complex EGFR mutations still exist, data of which are scarce and controversial. Hence, their role in response to standard therapy remains uncertain. We present the case of a patient diagnosed with stage IV lung adenocarcinoma for whom standard chemotherapies, including platinum agents, had failed. The patient was found to have an EGFR exon 19 (L747P) mutation, as evident in her liquid biopsy. This alteration has not been described before in the literature on non-Asian females. Data from the current case study highlight the aggressive nature of this type of EGFR mutation as indicated by the complete resistance to erlotinib. Using standard first-generation EGFR inhibitors in treating this point mutation was considered inadequate. However, this patient showed a substantial response when treated with erlotinib combined with epigenetic therapies, consisting of DNA methyltransferase and histone deacetylase inhibitors. For more than 8 years, the patient has been responding to combination therapy with a normal quality of life. This case represents a possible novel approach to reducing resistance in patients harboring this rare EGFR mutation which may translate to better outcomes.


A "new" chromosome marker common to the Rett syndrome and infantile autism? The frequency of fragile sites at X p22 in 81 children with infantile autism, childhood psychosis and the Rett syndrome.
Chromosomes from 46 autistic, 20 psychotic and 15 Rett syndrome children were cultured in a folic-acid-depleted medium. Nine percent of the autistic, 20% of the psychotic and 40% of the Rett syndrome cases showed a "new" chromosomal anomaly, viz a fragile site at the (X) (p22) location. It is suggested that in some cases of autism/psychosis and the Rett syndrome, there might be a common biological marker for the common type of psychiatric disturbance. However, as the population frequency of the chromosome marker is not yet known, conclusions must be drawn with great caution.


Intervention in the aging immune system: Influence of dietary restriction, dehydroepiandrosterone, melatonin, and exercise.
The decline in immunologic function with age is associated with an increase in susceptibility to infections and the occurrence of autoimmune diseases and cancers. Hence, the restoration of immunologic function is expected to have a beneficial effect in reducing pathology and maintaining a healthy condition in advanced age. A number of therapeutic strategies have been employed to intervene in the aging immune system. This article reviews the effect of dietary restriction (DR), dehydroepiandrosterone (DHEA) treatment, melatonin (MLT) therapy, and exercise on modulating the immune responses and retarding/reducing immunosenescence. DR has been subject to intensive research and is known to be the most efficacious means of increasing longevity, reducing pathology and enhancing immune function. The circulatory levels of the androgenic hormone DHEA and the pineal hormone MLT decrease with increasing age, and this decrease has been correlated with the age-related decline in the immune system. Therefore, the observation that immunosenescence is associated with low levels of DHEA and MLT has provided a rationale for therapeutic intervention. DHEA treatment and MLT therapy both exhibit immunostimulatory actions and preliminary reports indicate that hormonal (DHEA or MLT) substitution therapy reverses immunosenescence in mice. Similarly, exercise in some studies has been shown to enhance the immune response. However, these findings have not been confirmed by other laboratories. Thus, at the present time, it is difficult to draw any definitive conclusions on the efficacy of DHEA, MLT, and exercise on reversing or restoring the aging immune system.


Electrofusion of mouse blastomeres.
Fusion of blastomeres of 2-cell mouse embryos with an intact zona pellucida can be induced with electric pulses. Fusion was most frequent with the field strength of 1 kV/cm and direct current pulses of 100-250 microsec duration. An electrolyte solution (PBS) can be used instead of a non-electrolyte solution (0.3 M mannitol). The viability of blastomeres fused in these two types of solution is similar. Fused 2-cell blastomeres develop into tetraploid blastocysts but die after implantation. Embryos in which blastomeres failed to fuse despite the treatment (diploid controls) can develop till term. The technique can also be applied to 3- and 4-cell embryos and to zona-free oocytes and blastomeres.


Differential type 1 interferon-regulated gene expression in the brain during AIDS: interactions with viral diversity and neurovirulence.
The lentiviruses, human and feline immunodeficiency viruses (HIV-1 and FIV, respectively), infect the brain and cause neurovirulence, evident as neuronal injury, inflammation, and neurobehavioral abnormalities with diminished survival. Herein, different lentivirus infections in conjunction with neural cell viability were investigated, concentrating on type 1 interferon-regulated pathways. Transcriptomic network analyses showed a preponderance of genes involved in type 1 interferon signaling, which was verified by increased expression of the type 1 interferon-associated genes, Mx1 and CD317, in brains from HIV-infected persons (P<0.05). Leukocytes infected with different strains of FIV or HIV-1 showed differential Mx1 and CD317 expression (P<0.05). In vivo studies of animals infected with the FIV strains, FIV(ch) or FIV(ncsu), revealed that FIV(ch)-infected animals displayed deficits in memory and motor speed compared with the FIV(ncsu)- and mock-infected groups (P<0.05). TNF-α, IL-1β, and CD40 expression was increased in the brains of FIV(ch)-infected animals; conversely, Mx1 and CD317 transcript levels were increased in the brains of FIV(ncsu)-infected animals, principally in microglia (P<0.05). Gliosis and neuronal loss were evident among FIV(ch)-infected animals compared with mock- and FIV(ncsu)-infected animals (P<0.05). Lentiviral infections induce type 1 interferon-regulated gene expression in microglia in a viral diversity-dependent manner, representing a mechanism by which immune responses might be exploited to limit neurovirulence.


Role of tympanometric volume in paediatric tympanoplasty.
To determine the prognostic significance of "Tympaometric Volume" for paediatric tympanoplasty type I in a select age-group of 5-8 years. A prospective study was conducted in 30 children with chronic suppurative otitis media-inactive mucosal disease of either sex. Pre-operative tympanometric volume was recorded in all the cases and statistically analysed with the graft uptake results post-operatively. All the patients underwent tympanoplasty type I by underlay technique using temporalis fascia graft. An intact graft at the end of 6 months, and a postoperative hearing improvement of 10 dB or greater in two consecutive frequencies, was regarded as surgical and audiological success, respectively. The statistical analysis was done using Mantel Haenszel χ(2) i.e. Chi square test, and Fisher exact p value test for confirmation. We recorded an impressive surgical success rate of 87% and an audiological improvement of 70% in this study. On the basis of mean tympanometric volume of 1.6 cm(3) the patients were divided into two groups: in Group A (tympanometric volume<1.6 cm(3)), and group B (tympanometric volume>1.6 cm(3)). A graft uptake of 95% and 77% was recorded in Group A & B, respectively. However, the statistical evaluation of the data revealed no significant effect of this factor. In this study no correlation between the tympanometric volume and the surgical success of paediatric tympanoplasty in select age group of 5-8 years was observed.


[Postoperative T cell suppression in relation to preoperative nutritional status in childhood].
The influence of moderate malnutrition on the cellular immunity was studied in 59 operated infants and children aged 1 day to 14 years. Except during the neonatal period the number of blood T-cells preoperatively and the degree of the postoperative T-cell suppression did not show a significant difference between children suffering from moderate malnutrition (weight for age less than or equal to 10 percent, less than or equal to 90% of the normal median weight) and the control group. However some datas indicate that even borderline malnutrition impairs the cellular immune-status.


Salud Sí: a case study for the use of participatory evaluation in creating effective and sustainable community-based health promotion.
Participatory evaluation can be an essential tool for community-based organizations in tailoring programs to the needs of the populations they serve. This article provides a case study of Salud Sí, a promotora-driven health promotion program designed to encourage physical activity, fruit and vegetable consumption, and stress reduction among Mexican American women. Through a partnership between a community health center and an academic institution, we describe how the participatory evaluation framework is applied over a 10-year period throughout the stages of program development, implementation, and sustainability. Partners used the results to identify the essential elements of the health promotion program.


A new species of Deinodryinus Perkins, 1907 (Hymenoptera, Dryinidae) from Minas Gerais, Brazil.
Deinodryinus pratapolensis sp. nov. is described and illustrated. This species was collected by yellow pan trap, in a gallery forest at Pratápolis (Minas Gerais, Brazil).


Gross yield driving the mass fluxes of fishery drugs: Evidence of occurrence from full aquaculture cycle in lower Yangtze River Basin.
Expanding aquaculture has generated pollutants like fishery drugs in wastewater, which affects the aquatic environments and hinders sustainable development of aquaculture. To evaluate the occurrence, mass fluxes and production factors of fishery drugs in aquaculture, full-aquaculture-cycle monitoring in finfish and crustacean wastewater was conducted in the lower Yangtze River Basin, and 28 pesticides and 15 antibiotics were detected. The results showed that individual fishery drugs varied from ppt to ppb levels. Among them, sulfonamides were dominant with a mean concentration of 105.95 ± 4.13 ng·L-1 in finfish aquacultural wastewater, and insecticides were prevailing in crustacean aquacultural wastewater with a content of 146.56 ± 0.66 ng·L-1. Since the susceptibility to finfish disease determined the aquaculture practice, there were significant differences between two types of aquacultural wastewater. Finfish aquacultural wastewater contained more drugs and reached peak earlier in rapid-growth period, yet crustacean aquacultural wastewater peaked at the harvest period, to prevent against disease. Meanwhile, higher ecological risk, especially for florfenicol, were found in finfish wastewater. With 6 production factors from Good Aquaculture Practice, the gross yield was the most influential factor of drug mass flux, explaining 98 % variance by stepwise regression. Apart from increasing concentrations of fishery drugs in wastewater, regional high-yield aquaculture also significantly impacted the corresponding mass flux. As estimated by linear regression, 1.63 tons of target drugs would be discharged by 1 Mt. aquatic products, and 7.77 tons were discharged from aquaculture in the lower Yangtze River Basin in 2021. This is the first report to quantify mass fluxes of fishery drugs and to highlight gross yield as the most influential factor, which provides guidance for the supervision and regulation of sustainable aquaculture.


Effect of accelerated natural lactic fermentation of infant food ingredients on some pathogenic microorganisms.
Accelerated natural lactic fermentation in mixtures of water and ground sorghum, millet and pigeon pea was obtained by gradual selection of lactic acid bacteria, through inoculum recycling. Weaning food prepared from ingredients fermented this way, contained approx. 0.7% lactic and 0.05% acetic acids and had a pH of about 3.8. In porridges, a pH of less than or equal to 4.0 was required to cause death of Salmonella typhimurium and Staphylococcus aureus. Several intestinal pathogenic bacteria were inoculated into sour porridge. The most resistant Salmonella sp. died at a rate of 1.2 log cycle/h; the most resistant Shigella sp. at 0.9 log cycle/h; and the most resistant Escherichia coli strain at 0.6 log cycle/h. A yeast, Candida albicans, could grow well in the sour product, whereas a bacteriophage (MS-2) was inactivated at a rate of 0.1 log cycle/h. In the acid-sensitive bacterial cultures, no gradual adaptation to acid environments could be observed. The survival studies were carried out at 30 degrees C.


BRF1 accelerates prostate tumourigenesis and perturbs immune infiltration.
BRF1 is a rate-limiting factor for RNA Polymerase III-mediated transcription and is elevated in numerous cancers. Here, we report that elevated levels of BRF1 associate with poor prognosis in human prostate cancer. In vitro studies in human prostate cancer cell lines demonstrated that transient overexpression of BRF1 increased cell proliferation whereas the transient downregulation of BRF1 reduced proliferation and mediated cell cycle arrest. Consistent with our clinical observations, BRF1 overexpression in a Pten-deficient mouse (PtenΔ/Δ BRF1Tg) prostate cancer model accelerated prostate carcinogenesis and shortened survival. In PtenΔ/Δ BRF1Tg tumours, immune and inflammatory processes were altered, with reduced tumoral infiltration of neutrophils and CD4 positive T cells, which can be explained by decreased levels of complement factor D (CFD) and C7 components of the complement cascade, an innate immune pathway that influences the adaptive immune response. We tested if the secretome was involved in BRF1-driven tumorigenesis. Unbiased proteomic analysis on BRF1-overexpresing PC3 cells confirmed reduced levels of CFD in the secretome, implicating the complement system in prostate carcinogenesis. We further identify that expression of C7 significantly correlates with expression of CD4 and has the potential to alter clinical outcome in human prostate cancer, where low levels of C7 associate with poorer prognosis.


Psychological test results at three stages of inpatient alcoholism treatment.
Improvement in intellectual, perceptual-motor and personality functioning was observed in men alcoholics after 30 and 60 days of treatment. On the basis of results with the Minnesota Multiphasic Personality Inventory, four consistent personality clusters were identified.


Stereocontrolled synthesis of (E,Z)-dienals via tandem Rh(I)-catalyzed rearrangement of propargyl vinyl ethers.
A novel Rh(I)-catalyzed approach to functionalized (E,Z) dienals has been developed via tandem transformation where a stereoselective hydrogen transfer follows a propargyl Claisen rearrangement. Z-Stereochemistry of the first double bond suggests the involvement of a six-membered cyclic intermediate whereas the E-stereochemistry of the second double bond stems from the subsequent protodemetalation step giving an (E,Z)-dienal.


Endovascular treatment of intracranial aneurysms.
To examine the techniques, reported experiences, and advantages and disadvantages associated with the endovascular treatment of intracranial aneurysms. We review the endovascular techniques used for the treatment of intracranial aneurysms and the sequelae of subarachnoid hemorrhage, which have evolved during the past 10 years. Two broad categories of endovascular therapy for intracranial aneurysms are described: occlusion of the parent artery and preservation of the parent artery by selective occlusion of the aneurysm with balloons or metallic coils. The Mayo protocol for testing tolerance of patients before permanent balloon occlusion of the parent artery is described, as are the types of aneurysms most amenable to this treatment. In addition, use of balloon angioplasty for cerebral vasospasm after subarachnoid hemorrhage is reviewed. Recent improvements in microcatheter technology have facilitated the safe navigation of percutaneously introduced catheters in the intracranial circulation and selective catheterization of intracranial aneurysms. Surgically difficult aneurysms are now being treated with endovascular techniques more frequently than in the past. Early results from animal experiments and human trials have shown that selective occlusion of aneurysms with metallic coils may have a role in the treatment of intracranial aneurysms. Balloon angioplasty of symptomatic cerebral vasospasm has demonstrated improvement in neurologic function in approximately 70% of patients. As technology continues to improve and as greater experience is obtained, interventional neuroradiologists will continue to have an increasingly important role in the treatment of intracranial aneurysms.


Learning styles differ between senior dietetics students and dietetics faculty members.
This study examined learning styles of faculty members and senior students in coordinated programs in dietetics and compared learning styles by area of career expertise (faculty) or interest (students). Respondents included 108 faculty members and 324 students. The survey included a standardized Learning Styles Inventory and questions regarding demographics and career preferences. chi(2) analysis was used to compare learning style with selected variables. Significant differences were found between student and faculty learning styles (P=.006). Faculty also had significant differences in learning styles within various areas of expertise (P=.037), but no significant differences were found in learning style by expected career choice of students. Students listed entry-level and 5-year career choices. Most (55.6%) plan to work in clinical nutrition following graduation, but 37.0% anticipate being in a nontraditional area of dietetics 5 years postgraduation. Only 6.8% plan to work in management at entry, dropping to 3.4% at 5 years.


A previously unrecognized membrane protein in the Rhodobacter sphaeroides LH1-RC photocomplex.
Rhodobacter (Rba.) sphaeroides is the most widely used model organism in bacterial photosynthesis. The light-harvesting-reaction center (LH1-RC) core complex of this purple phototroph is characterized by the co-existence of monomeric and dimeric forms, the presence of the protein PufX, and approximately two carotenoids per LH1 αβ-polypeptides. Despite many efforts, structures of the Rba. sphaeroides LH1-RC have not been obtained at high resolutions. Here we report a cryo-EM structure of the monomeric LH1-RC from Rba. sphaeroides strain IL106 at 2.9 Å resolution. The LH1 complex forms a C-shaped structure composed of 14 αβ-polypeptides around the RC with a large ring opening. From the cryo-EM density map, a previously unrecognized integral membrane protein, referred to as protein-U, was identified. Protein-U has a U-shaped conformation near the LH1-ring opening and was annotated as a hypothetical protein in the Rba. sphaeroides genome. Deletion of protein-U resulted in a mutant strain that expressed a much-reduced amount of the dimeric LH1-RC, indicating an important role for protein-U in dimerization of the LH1-RC complex. PufX was located opposite protein-U on the LH1-ring opening, and both its position and conformation differed from that of previous reports of dimeric LH1-RC structures obtained at low-resolution. Twenty-six molecules of the carotenoid spheroidene arranged in two distinct configurations were resolved in the Rba. sphaeroides LH1 and were positioned within the complex to block its channels. Our findings offer an exciting new view of the core photocomplex of Rba. sphaeroides and the connections between structure and function in bacterial photocomplexes in general.


The use of biolistic inoculation of cassava mosaic begomoviruses in screening cassava for resistance to cassava mosaic disease.
Inoculation of cassava with infectious clones of cassava mosaic geminiviruses (Geminiviridae: Begomovirus) and total DNA extracts from plants infected with well-characterised viruses was evaluated using the Bio-Rad Helios Gene Gun System. Total DNA extracts from infected plants and cloned viruses were produced for coating gold particles and bombardment onto new cassava genotypes, 96/1089A, 96/1039, 96/0160, 96/0304 and three local landraces TME 117, TME 3 and TME 4. Cloned DNA of a Kenyan isolate of the recombinant variant of East African cassava mosaic virus (EACMV-UG2-[Ka]), was only infectious to TME 117 (7/10 plants), 3 weeks post-inoculation with mild infection symptoms in the newly developing leaves. Biolistic inoculation with a chimeric pseudorecombinant virus between DNA A and B components from EACMV-[Ke-Kilifi] and EACMV-UG2-[Ka], respectively, was infectious to TME 117, 96/1039 and 96/0304 and developed very severe and persistent symptoms. TME 3 and TME 4 also developed symptoms, 12 days post-inoculation (d.p.i.). Total DNA extracts of ACMV and EACMV-[Ke-Kilifi] resulted in serious infections with symptoms already evident, 10d.p.i. In general, biolistic inoculation trials with total DNA extracts resulted in a higher number of infected plants expressing symptoms at a much earlier stage (10-12d.p.i.) compared with trials inoculated with virus clones.


A randomized trial to assess catheter ablation versus rate control in the management of persistent atrial fibrillation in heart failure.
This study sought to compare catheter ablation with rate control for persistent atrial fibrillation (AF) in heart failure (HF). The optimal therapy for AF in HF is unclear. Drug-based rhythm control has not proved clinically beneficial. Catheter ablation improves cardiac function in patients with HF, but impact on physiological performance has not been formally evaluated in a randomized trial. In a randomized, open-label, blinded-endpoint clinical trial, adults with symptomatic HF, radionuclide left ventricular ejection fraction (EF) ≤35%, and persistent AF were assigned to undergo catheter ablation or rate control. Primary outcome was 12-month change in peak oxygen consumption. Secondary endpoints were quality of life, B-type natriuretic peptide, 6-min walk distance, and EF. Results were analyzed by intention-to-treat. Fifty-two patients (age 63 ± 9 years, EF 24 ± 8%) were randomized, 26 each to ablation and rate control. At 12 months, 88% of ablation patients maintained sinus rhythm (single-procedure success 68%). Under rate control, rate criteria were achieved in 96%. The primary endpoint, peak oxygen consumption, significantly increased in the ablation arm compared with rate control (difference +3.07 ml/kg/min, 95% confidence interval: 0.56 to 5.59, p = 0.018). The change was not evident at 3 months (+0.79 ml/kg/min, 95% confidence interval: -1.01 to 2.60, p = 0.38). Ablation improved Minnesota score (p = 0.019) and B-type natriuretic peptide (p = 0.045) and showed nonsignificant trends toward improved 6-min walk distance (p = 0.095) and EF (p = 0.055). This first randomized trial of ablation versus rate control to focus on objective exercise performance in AF and HF shows significant benefit from ablation, a strategy that also improves symptoms and neurohormonal status. The effects develop over 12 months, consistent with progressive amelioration of the HF syndrome. (A Randomised Trial to Assess Catheter Ablation Versus Rate Control in the Management of Persistent Atrial Fibrillation in Chronic Heart Failure; NCT00878384).


Reciprocal regulation of sex-dependent expression of testosterone 15 alpha-hydroxylase (P-450(15 alpha)) in liver and kidney of male mice by androgen. Evidence for a single gene.
Testosterone 15 alpha-hydroxylase activities and its mRNA levels are higher in kidneys than in livers from male 129/J mice. Castration of 129/J male mice resulted in repression of P-450(15 alpha) in kidney, but increased it in liver. Two types of cDNA (p15 alpha-29 (Type I) and -15 (Type II)) encoding P-450(15 alpha) were previously cloned from 129/J female livers (Burkhart, B.A., Harada, N., and Negishi, M. (1985) J. Biol. Chem. 260, 15357-15361). With the use of p15 alpha-29 as a probe, Type I and II P-450(15 alpha) cDNAs were isolated from libraries of 129/J kidney poly(A)+ RNA. The nucleotide sequences of the cDNAs showed that Type I and II cDNAs from liver and kidney were identical and shared 98.3% similarity. The deduced amino acid sequence from a full-length Type I cDNA indicated that Type I P-450(15 alpha) consists of 494 amino acids with a molecular weight of 56,594. Nine amino acid substitutions were found in the Type II clone in 432 amino acids overlapping Type I. Type I cDNA clones accounted for approximately 90% P-450(15 alpha) clones isolated from a male kidney library, whereas approximately 90% of cDNA clones in a female kidney library were Type II. Liver cDNA libraries from males and females contained similar ratios of Type I and II. Effects of castration on Type I and II mRNAs were determined by Southern hybridization of a 32P-labeled ClaI-ClaI fragment from p15 alpha-29 to cDNAs synthesized from kidney and liver poly(A)+ RNAs prepared from sham-operated, castrated 129/J mice. The double-stranded cDNAs were digested with ClaI and PstI prior to gel electrophoresis to create the diagnostic restriction fragments specific for Type I or II. Castration resulted in decreased levels of Type I mRNA in male kidney. In male liver, only Type I mRNA rose significantly in response to castration. Testosterone administration returned the Type I mRNA to normal levels in castrated mice. It therefore appears that the high levels of P-450(15 alpha) in male kidney were due to androgen-dependent induction of Type I mRNA. Both Types I and II were repressed in male liver, which results in decreased levels of P-450(15 alpha). Androgen was responsible for the repression and expression of Type I in liver and kidney, but not Type II.


Oscillations and variability in neuronal systems: interplay of autonomous transient dynamics and fast deterministic fluctuations.
Neuronal systems are subject to rapid fluctuations both intrinsically and externally. These fluctuations can be disruptive or constructive. We investigate the dynamic mechanisms underlying the interactions between rapidly fluctuating signals and the intrinsic properties of the target cells to produce variable and/or coherent responses. We use linearized and non-linear conductance-based models and piecewise constant (PWC) inputs with short duration pieces. The amplitude distributions of the constant pieces consist of arbitrary permutations of a baseline PWC function. In each trial within a given protocol we use one of these permutations and each protocol consists of a subset of all possible permutations, which is the only source of uncertainty in the protocol. We show that sustained oscillatory behavior can be generated in response to various forms of PWC inputs independently of whether the stable equilibria of the corresponding unperturbed systems are foci or nodes. The oscillatory voltage responses are amplified by the model nonlinearities and attenuated for conductance-based PWC inputs as compared to current-based PWC inputs, consistent with previous theoretical and experimental work. In addition, the voltage responses to PWC inputs exhibited variability across trials, which is reminiscent of the variability generated by stochastic noise (e.g., Gaussian white noise). Our analysis demonstrates that both oscillations and variability are the result of the interaction between the PWC input and the target cell's autonomous transient dynamics with little to no contribution from the dynamics in vicinities of the steady-state, and do not require input stochasticity.


Increased incidence of metabolic syndrome among older survivors relocated to temporary housing after the 2011 Great East Japan earthquake & tsunami.
Beyond the immediate toll of injuries and deaths, major disasters are often associated with long-term increased risks of chronic disease. We sought to investigate the incidence of metabolic syndrome (MetS) among survivors of the 2011 Great East Japan Earthquake and tsunami. Subjects aged ≥18 years from the tsunami-stricken area participated in a prospective cohort study of disaster survivors (the RIAS Study) from 2011 to 2015. After excluding subjects who were previously diagnosed with MetS, we observed the cumulative incidence of MetS across four annual examinations among 7318 subjects (mean age, 59.8 years; 43.5% men). We defined MetS using the International Diabetes Foundation criteria. The 4-year cumulative incidence of MetS was 18.0% in the overall sample. The incidence was significantly higher among older women survivors relocated to prefabricated temporary housing (40.9%, 95% confidence interval, 36.4-44.6), and other types of housing (36.2%, 95% CI: 32.3-40.6) compared to those who were not relocated (34.1%, 95% CI: 30.9-37.4). An increase in incidence of MetS was not observed for older men, or younger survivors aged ≤64 years. Relocation to prefabricated temporary housing was a risk factor for increased incidence of MetS in older women.


Effect of splenectomy and immunotherapy on advanced gastric cancer associated with total gastrectomy.
Immunotherapy with levamisole (LMS) and combined splenectomy were performed in the cases of stages III and IV gastric cancer of the upper and middle stomach and associated with total gastrectomy. The cell-mediated immunity and the survival rates for both groups were compared with controls. Combined splenectomy was not effective for cases of stage III gastric cancer, but was effective for those in stage IV. Comparing the cumulative survival rates of the immunotherapy group with the controls, cases of stages III and IV gastric cancer without splenectomy, the survival rate of those with immunotherapy was higher than that of the control group in the cases of the stage III, but the result was reverse in cases of stage IV gastric cancer. Changes in the cell-mediated immunity correlated with the survival rates.