A Monoclonal Antibody that Binds Preferentially to Human Suppressor T Cells.
A monoclonal antibody termed JM3-3-3A was produced by somatic cell hybridization. The reactivity was assessed by indirect immunofluorescence. JM3-3-3A was reactive with 41% human peripheral blood T cells and 48% non-T cells. Among various human lymphoblastoid cell lines (MOLT 4F, JM, and TALL-I), TALL-I was found not to be reactive with JM-13-3-3A. Human peripheral blood Tcells fractionated by JM3-3-3A-coated dish panning were submitted to functional studies. JM3-3-3A positive T cells responded to mitogens, concanavalin A and phytohemagglutinin-P much better than JM3-3-3A negative T cells in the presence or absence of adherent cells. JM3-3-3A positive T cells showed suppressor activity and JM3-3-3A negative T cells showed helper activity in pokeweed mitogen-induced Ig production. More than fifty percent of the JM3-3-3A positive T cells were reactive with OKT8 which binds to suppressor/ cytotoxic T cells, whereas 18% of JM3-3-3A negative T cells were reactive with OKTIl. In addition, immunoprecipitation experiments identified a protein with an approximate molecular weight of 43,000 as a cell surface antigen for JM3-3-3A. Thus, the reactivity of JM3-3-3A showed a wide distribution but human peripheral blood T cells could be dissected functionally by this antibody.


Assessment of DFT approaches in noble gas clathrate-like clusters: stability and thermodynamics.
We have assessed the performance and accuracy of different wavefunction-based electronic structure methods, such as DFMP2 and domain-based local pair-natural orbital (DLPNO-CCSD(T)), as well as a variety of density functional theory (DFT) approaches on He@(H2O)N cage systems. We have selected representative clathrate-like structures corresponding to the building blocks present in each of the sI, sII and sH natural gas clathrate hydrates, and we have carefully studied the interaction between a He atom with each of their individual cages. We reported well-converged DFMP2 and DLPNO-CCSD(T) reference data, together with interaction and cohesive energies of four different density functionals (two GGA, revPBE and PW86PBE, and two hybrids, B3LYP and PBE0), including diverse dispersion correction schemes (D3(0), D3(BJ), D4 and XDM) for both He-filled and empty clathrate-like cages. After the analysis of the results, we came to the conclusion that the PW86PBE functional, with both XDM and D4 corrections, and the PBE0-D4 functional present reasonably adequate approaches to describe the guest-host noncovalent interactions that take place in such He clathrate hydrates. Taking into account that the He@sII is the only helium clathrate that scientists have been able to synthesize recently, we have performed a thermodynamic study on the individual 512 and 51264 cages present in the sII crystal. We determined the change in enthalpy, ΔH, and in Gibbs free energy, ΔG, at various temperatures and pressures, and we found out that in the range of experimental conditions the reactions associated with the encapsulation of the He atom inside the cages are exothermic and spontaneous. Finally, we highlighted the importance of an accurate description of the interaction in He@water mixtures, as a crucial component in construction of reliable data-driven models.


Cone-beam computed tomography: Time to move from ALARA to ALADA.
Cone-beam computed tomography (CBCT) is routinely recommended for dental diagnosis and treatment planning. CBCT exposes patients to less radiation than does conventional CT. Still, lack of proper education among dentists and specialists is resulting in improper referral for CBCT. In addition, aiming to generate high-quality images, operators may increase the radiation dose, which can expose the patient to unnecessary risk. This letter advocates appropriate radiation dosing during CBCT to the benefit of both patients and dentists, and supports moving from the concept of "as low as reasonably achievable" (ALARA) to "as low as diagnostically acceptable" (ALADA).


Lived experience narratives in health professional education: educators' perspectives of a co-designed, online mental health education resource.
Meaningful involvement of people with lived experience is an invaluable approach to education that facilitates the development of knowledge, skills and attitudes for collaborative, compassionate and person-centred healthcare practice. The purpose of this evaluation was to gain health professional educators' perspectives of an online learning resource that presents the lived experiences of people who have been consumers of the Australian mental health system. A cross sectional study design was used to survey educators who had registered to use the online education resource. Data were collected using an online survey and follow-up interviews. Two lived experience researchers were involved in the research. Quantitative survey data were analysed descriptively, and qualitative data were analysed thematically. The Listening to Voices online education resource is being used in a range of settings. Educators perceived the content facilitated achievement of learning outcomes related to understanding the experiences of people with mental health issues. The free, online, and flexible design of the resource promoted access and helped overcome barriers to including lived experience experts in education. The powerful impact of the resource and importance of creating safe learning environments when using the resource were highlighted. Suggestions for future developments were provided. Involving people with lived experience in education of healthcare students and professionals can assist in developing skills for collaborative, compassionate, and person-centred care. Implementation of co-design principles and the use of creative pedagogical approaches can contribute to the development of impactful educational resources that foreground lived experience. Making these resources flexible and freely available online improves their utility.


Phenotypic engineering by reprogramming gene transcription using novel artificial transcription factors in Escherichia coli.
Now that many genomes have been sequenced and the products of newly identified genes have been annotated, the next goal is to engineer the desired phenotypes in organisms of interest. For the phenotypic engineering of microorganisms, we have developed novel artificial transcription factors (ATFs) capable of reprogramming innate gene expression circuits in Escherichia coli. These ATFs are composed of zinc finger (ZF) DNA-binding proteins, with distinct specificities, fused to an E. coli cyclic AMP receptor protein (CRP). By randomly assembling 40 different types of ZFs, we have constructed more than 6.4 x 10(4) ATFs that consist of 3 ZF DNA-binding domains and a CRP effector domain. Using these ATFs, we induced various phenotypic changes in E. coli and selected for industrially important traits, such as resistance to heat shock, osmotic pressure and cold shock. Genes associated with the heat-shock resistance phenotype were then characterized. These results and the general applicability of this platform clearly indicate that novel ATFs are powerful tools for the phenotypic engineering of microorganisms and can facilitate microbial functional genomic studies.


Depletion of 14-3-3 zeta elicits endoplasmic reticulum stress and cell death, and increases vulnerability to kainate-induced injury in mouse hippocampal cultures.
14-3-3 proteins are ubiquitous signalling molecules that regulate development and survival pathways in brain. Altered expression and cellular localization of 14-3-3 proteins has been implicated in neurodegenerative diseases and in neuronal death after acute neurological insults, including seizures. Presently, we examined expression and function of 14-3-3 isoforms in vitro using mouse organotypic hippocampal cultures. Treatment of cultures with the endoplasmic reticulum (ER) stressor tunicamycin caused an increase in levels of 14-3-3 zeta within the ER-containing microsomal fraction, along with up-regulation of Lys-Asp-Glu-Leu-containing proteins and calnexin, and the selective death of dentate granule cells. Depletion of 14-3-3 zeta levels using small interfering RNA induced both ER stress proteins and death of granule cells. Treatment of hippocampal cultures with the excitotoxin kainic acid increased levels of Lys-Asp-Glu-Leu-containing proteins and microsomal 14-3-3 zeta levels and caused cell death within the CA1, CA3 and dentate gyrus of the hippocampus. Kainic acid-induced damage was significantly increased in each hippocampal subfield of cultures treated with small interfering RNA targeting 14-3-3 zeta. The present data indicate a role for 14-3-3 zeta in survival responses following ER stress and possibly protection against seizure injury to the hippocampus.


The use of instruments by Iranian endodontics and general practioners.
The current clinical practice of endodontics involves utilization of a variety of new technological advances and materials. Technologies available for use in endodontic offices today include battery or electric motors using NiTi rotary file systems, new generation of electronic apex locators (EALs), improved digital radiographic sensors, surgical microscopes, and ultrasonic units. The aim of this study was to determine the prevalence and use of newer technologies among Iranian dental practitioners. The population under study included the dental practitioners participating in the 49th and 50(th) Congress of Dentistry, who were selected with a simple sampling method. Data was collected by questionnaires which were completed by the participants in an anonymous manner. The contingency table and chi-squared test were used for data analysis by SPSS 13.5 software. A total of 700 dental practitioners participated in the study; NiTi rotary systems were used by 50.1%; electronic apex locators were often employed by 46.3%; 21.7% reported that ultrasonic units were often used. Only 1.1% frequently used surgical microscopes. Males and females differed with respect to the use of newer technologies (P<0.05). Based on the results of the present study, it seems necessary for dentists to take part in continuing dental education programs related to the newer technologies to improve their knowledge and practice.


Natural history, evaluation, and management of intracranial vascular malformations.
Intracranial vascular malformations are seen increasingly in clinical practice, primarily because of advances in cross-sectional brain and spinal cord imaging. Commonly encountered lesion types include arteriovenous malformations, cavernous malformations, venous malformations, dural arteriovenous fistulas, and capillary telangiectasias. Patients can experience various symptoms and signs at presentation. The natural history of vascular malformations depends on lesion type, location, size, and overall hemodynamics. The natural history for each lesion subtype is reviewed, with special consideration of the risk of hemorrhage or other adverse outcomes after the lesion is detected and any known predictors of hemorrhage or other outcomes. In practice, these data are compared with the risk of available treatment options as the optimal management is clarified. A multidisciplinary approach including neurosurgery, radiosurgery, interventional neuroradiology, and vascular neurology is most useful in determining the best management strategy.


Approaching the Traveling Tournament Problem with Randomized Beam Search.
The traveling tournament problem is a well-known sports league scheduling problem famous for its practical hardness. Given an even number of teams with symmetric distances between their venues, a double round-robin tournament has to be scheduled minimizing the total travel distances over all teams. We consider the most common constrained variant without repeaters and a streak limit of three, for which we study a beam search approach based on a state-space formulation guided by heuristics derived from different lower bound variants. We solve the arising capacitated vehicle routing subproblems either exactly for small- to medium-sized instances up to 18 teams or heuristically also for larger instances up to 24 teams. In a randomized variant of the search, we employ random team ordering and add small amounts of Gaussian noise to the nodes' guidance for diversification when multiple runs are performed. This allows for a simple yet effective parallelization of the beam search. A final comparison is done on the NL, CIRC, NFL, and GALAXY benchmark instances with 12 to 24 teams, for which we report a mean gap difference to the best known feasible solutions of 1.2% and five new best feasible solutions.


Delta-sleep inducing peptide entrapment in the charged macroporous matrices.
Various biomolecules, for example proteins, peptides etc., entrapped in polymer matrices, impact interactions between matrix and cells, including stimulation of cell adhesion and proliferation. Delta-sleep inducing peptide (DSIP) possesses numerous beneficial properties, including its abilities in burn treatment and neuronal protection. DSIP entrapment in two macroporous polymer matrices based on copolymer of dimethylaminoethyl methacrylate and methylen-bis-acrylamide (Co-DMAEMA-MBAA) and copolymer of acrylic acid and methylen-bis-acrylamide (Co-AA-MBAA) has been studied. Quite 100% of DSIP has been entrapped into positively charged Co-DMAEMA-MBAA matrix, while the quantity of DSIP adsorbed on negatively charged Co-AA-MBAA was only 2-6%. DSIP release from Co-DMAEMA-MBAA was observed in saline solutions (0.9% NaCl and PBS) while there was no DSIP release in water or 25% ethanol, thus ionic strength was a reason of this process.


Does Early Chemotherapy Improve Survival in Advanced Endometrial Cancer?
In endometrial cancer, the appropriate sequence of adjuvant chemotherapy (aCT) and adjuvant radiation therapy (aRT) is unclear. We aim evaluated whether early chemotherapy is associated with improved overall survival (OS) and cancer-specific survival (CSS). Endometrial cancer patients that received aCT and aRT were selected from the SEER-Medicare database. Early chemotherapy was defined as receiving aCT before aRT, with or without additional aCT ("sandwich" regimens). All other patients received a full course of aRT before chemotherapy with or without concurrent chemotherapy. Univariate and multivariate Cox proportional hazards regression was utilized to assess the impact of clinical and demographic factors on OS. We selected 597 patients for analysis. Median age and was 72 years; 85% of patients were white. Overall, 68% of women had FIGO (International Federation of Gynecology and Obstetrics) stage III disease and 77% received 4 to 6 cycles of chemotherapy. Five-year OS (66.6% vs. 62.4%, P=0.46) and 5-year CSS (71.1% vs. 71.2%, P=0.88) was not significantly improved among those receiving early chemotherapy. In addition, early chemotherapy did not improve OS (hazard ratio [HR]=0.87; 95% confidence interval [CI]: 0.56-1.34, P=0.53) or CSS (HR=1.21; 95% CI: 0.82-1.79, P=0.34) on multivariate analysis. Compared with 1 to 3 cycles, receiving 4 to 6 (HR=0.48, 95% CI: 0.26-0.87, P=0.02), and ≥7 cycles (HR=0.42, 95% CI: 0.20-0.89, P=0.02) of chemotherapy was associated with improved OS. No differences in OS or CSS were noted among endometrial patients receiving early chemotherapy. However, the number of chemotherapy cycles was associated with prolonged survival.


Absorption of ignitable liquids into polyethylene/polyvinylidine dichloride bags.
Clear plastic bags are often used for the collection, sampling and storage of ignitable liquid evidence. They are popular because they are easy to store. transport and are inexpensive. Cryovac and Globus brand polyethylene/polyvinylidine dichloride bags were tested for suitability in storing ignitable liquid evidence. Standards of diesel, kerosene and gasoline were placed in the bags and sampled by passive headspace adsorption. The bags were then heated to determine if absorbed components of the standards could be released upon heating. Recovered extracts were analyzed by GC and GCMS. These bags were found to absorb components of diesel, kerosene, and gasoline. and were also found to produce interfering by-products that obstruct the chromatographic results. Evidence containers need to be tested to ensure that low levels of ignitable liquids are not missed.


Acupuncture modulates cortical thickness and functional connectivity in knee osteoarthritis patients.
In this study, we investigated cortical thickness and functional connectivity across longitudinal acupuncture treatments in patients with knee osteoarthritis (OA). Over a period of four weeks (six treatments), we collected resting state functional magnetic resonance imaging (fMRI) scans from 30 patients before their first, third and sixth treatments. Clinical outcome showed a significantly greater Knee Injury and Osteoarthritis Outcome Score (KOOS) pain score (improvement) with verum acupuncture compared to the sham acupuncture. Longitudinal cortical thickness analysis showed that the cortical thickness at left posterior medial prefrontal cortex (pMPFC) decreased significantly in the sham group across treatment sessions as compared with verum group. Resting state functional connectivity (rsFC) analysis using the left pMPFC as a seed showed that after longitudinal treatments, the rsFC between the left pMPFC and the rostral anterior cingulate cortex (rACC), medial frontal pole (mFP) and periaquiduct grey (PAG) are significantly greater in the verum acupuncture group as compared with the sham group. Our results suggest that acupuncture may achieve its therapeutic effect on knee OA pain by preventing cortical thinning and decreases in functional connectivity in major pain related areas, therefore modulating pain in the descending pain modulatory pathway.


Challenges to tuberculin screening and follow-up in an urban Aboriginal sample in Montreal, Canada.
We sought to describe the prevalence and predictors of tuberculin skin test (TST) reactors in a high-risk sample of urban Aboriginal people, and to evaluate adherence to medical evaluation and latent tuberculosis infection (LTBI) treatment among those with TST reactivity. Of the 164 participants tested, 86% returned for TST reading. Positive TST reactions (>or=10 mm) were observed in 17.7% (25/141, 95% CI 11.4-24.0) of participants, and were associated with older age (OR per 10 year increase 1.8, 95% CI 1.2-2.7) and Inuit Aboriginal group (OR 2.8, 95% CI 1.1-7.3). Only four participants presented for evaluation, of whom one initiated and none completed LTBI treatment. Tuberculin screening in this population can be an effective strategy for identifying TST reactive individuals; however, screening efforts will have minimal impact without additional efforts in this high-risk group.


From microbiome composition to functional engineering, one step at a time.
SUMMARYCommunities of microorganisms (microbiota) are present in all habitats on Earth and are relevant for agriculture, health, and climate. Deciphering the mechanisms that determine microbiota dynamics and functioning within the context of their respective environments or hosts (the microbiomes) is crucially important. However, the sheer taxonomic, metabolic, functional, and spatial complexity of most microbiomes poses substantial challenges to advancing our knowledge of these mechanisms. While nucleic acid sequencing technologies can chart microbiota composition with high precision, we mostly lack information about the functional roles and interactions of each strain present in a given microbiome. This limits our ability to predict microbiome function in natural habitats and, in the case of dysfunction or dysbiosis, to redirect microbiomes onto stable paths. Here, we will discuss a systematic approach (dubbed the N+1/N-1 concept) to enable step-by-step dissection of microbiome assembly and functioning, as well as intervention procedures to introduce or eliminate one particular microbial strain at a time. The N+1/N-1 concept is informed by natural invasion events and selects culturable, genetically accessible microbes with well-annotated genomes to chart their proliferation or decline within defined synthetic and/or complex natural microbiota. This approach enables harnessing classical microbiological and diversity approaches, as well as omics tools and mathematical modeling to decipher the mechanisms underlying N+1/N-1 microbiota outcomes. Application of this concept further provides stepping stones and benchmarks for microbiome structure and function analyses and more complex microbiome intervention strategies.


Are Impulsive Decisions Always Irrational? An Experimental Investigation of Impulsive Decisions in the Domains of Gains and Losses.
Intertemporal choices are very prevalent in daily life, ranging from simple, mundane decisions to highly consequential decisions. In this context, thinking about the future and making sound decisions are crucial to promoting mental and physical health, as well as a financially sustainable lifestyle. In the present study, we set out to investigate some of the possible underlying mechanisms, such as cognitive factors and emotional states, that promote future-oriented decisions. In a cross-sectional experimental study, we used a gain and a loss version of an intertemporal monetary choices task. Our main behavioural result indicated that people are substantially more impulsive over smaller and sooner monetary losses compared to equivalent gains. In addition, for both decisional domains, significant individual difference predictors emerged, indicating that intertemporal choices are sensitive to the affective and cognitive parameters. By focusing on the cognitive and emotional individual factors that influence impulsive decisions, our study could constitute a building block for successful future intervention programs targeted at mental and physical health issues, including gambling behaviour.


eDNA monitoring as a tool for evaluating the reintroduction of Austropotamobius pallipes after a crayfish plague outbreak.
The crayfish plague, a severe disease caused by the oomycete Aphanomyces astaci, is responsible for most population declines of susceptible crayfish in Europe. This pathogen has been devastating native populations of Austropotamobius pallipes since the 1970s in the Iberian Peninsula. In this study, we report a massive mortality event in one of the most important Spanish populations of A. pallipes. We aimed to: (i) identify the cause of the mortality, and (ii) evaluate the reintroduction viability of the species. Over the course of six months, we used environmental DNA (eDNA) and traditional trap-based methods to detect the presence of A. astaci or of native or invasive crayfish in order to evaluate the reintroduction viability of A. pallipes to the affected population. We did not capture any live crayfish or detect the presence of A. astaci in the reservoir water during the six months following the mass mortality event. Our analyses indicated that it was feasible to initiate a reintroduction program at the site, which will continue to be monitored for three to five years and will help improve the conservation status of A. pallipes.


Silane-initiated nucleation in chemically active plasmas: validation of density functionals, mechanisms, and pressure-dependent variational transition state calculations.
The growth of anionic silicon hydride clusters is a critically important process in nanodusty plasmas. In the current study, we focus on the formation of homologs of silylene (Sin+1H2n+2(-), n = 3, 4) and silyl (SinH2n+1(-), n = 4, 5) anions via anion-neutral reaction pathways. Species like silyl or silylene anions and their related elementary reactions, which are involved in the formation of silicon hydride clusters, were not used in developing exchange-correlation (xc) density functionals (i.e., they were not included in the training set of semiempirical density functionals); therefore, we explored the accuracy of various widely used xc density functionals based on reaction energies and barrier heights. Among the 21 density functionals we tested, M06-2X has the best performance for a hybrid functional, and MN15-L has the best performance for a local functional. Thermal rate constants of the elementary reactions involved in the reaction mechanism are calculated using M06-2X and multistructural canonical variational transition state theory with the small-curvature tunneling approximation (MS-CVT/SCT). The pressure dependence of unimolecular isomerization reactions is treated with system-specific quantum RRK theory (SS-QRRK) and the Lindemann-Hinshelwood mechanism.


Atracurium, vecuronium, and intraocular pressure in humans.
We studied 60 nonophthalmologic patients, allocated to six treatment groups, to assess the effects of atracurium and vecuronium on intraocular pressure (IOP). All patients had IOP measured while awake, using pneumotonometry. In group 1, anesthesia was induced with thiopental, 5 mg/kg, and maintained with N2O, 70% in O2, using controlled mask ventilation, for 5 min. These patients then received atracurium, 0.5 mg/kg. After 5 additional minutes of ventilation, the trachea was intubated. From 1 min after thiopental administration until 1 min after intubation, IOP was recorded every minute. Patients in groups 2, 3, and 4 were treated identically to those in group 1, except the muscle relaxant given was atracurium, 1.0 mg/kg, vecuronium, 0.1 mg/kg, or vecuronium, 0.2 mg/kg, respectively. Patients in groups 5 and 6 underwent rapid sequence induction with thiopental, 5 mg/kg, and atracurium, 1.0 mg/kg, or vecuronium, 0.2 mg/kg, respectively. IOP was measured 1 min later, followed by intubation and IOP measurements for the next 3 min. Intraocular pressure decreased significantly in groups 1, 2, 4, and 6 after thiopental and remained stable in all groups during ventilation with N2O. Neither atracurium nor vecuronium affected IOP, nor was there any correlation between IOP and degree of neuromuscular blockade. However, IOP increased significantly after intubation in all six groups. We conclude that atracurium or vecuronium alone has no adverse effects on IOP.


Thin, lightweight, foldable thermochromic displays on paper.
This article describes an electronic display that is fabricated by patterning electrically conductive wires (heaters) with micron-scale dimensions on one side of a sheet of paper, and thermochromic ink on the opposite side. Passing electrical current through the wires heats the paper and changes the thermochromic ink from colored (black, green, or other colors) to transparent; this change in property reveals the paper underneath the ink-exposing any messages printed on the paper-and serves as the basis for a two-state "shutter" display. This type of display is thin (100 microm), flat, lightweight (the display weighs <20 mg/cm(2)), can be folded, rolled, twisted, and creased while maintaining function, and ultimately can (if required) be disposed of by incineration. The display is appropriate for applications where information must be presented clearly (usually only once) for little cost (each display costs <$0.10/m(2) in materials) and where limited electrical power is available.


Investigating the discernible and distinct effects of platinum-based chemotherapy regimens for metastatic triple-negative breast cancer on time to progression.
Platinum-based chemotherapy regimens are frequently used in patients with triple-negative breast cancer (TNBC). The aim of the current study was to assess whether or not platinum-based chemotherapy is associated with an increased time to progression when compared with non-platinum-based regimens in TNBC and non-TNBC. A retrospective analysis was conducted within a cohort of patients with metastatic breast cancer who received platinum-based chemotherapy at a single institution. Data were collected for up to three lines of treatment for metastatic disease. Time to progression was determined for platinum-based chemotherapy and non-platinum-based regimens for each line of treatment. Adjusted hazard ratios (HRs), together with 95% confidence intervals (CIs) were estimated comparing the time to progression associated with the use of platinum-based chemotherapy versus non-platinum-based regimens. A total of 159 patients were included in the analysis, with 58 diagnosed with TNBC. Among the patients with TNBC, compared with non-platinum-based chemotherapy, no correlation was identified between platinum-based chemotherapy and an improved time to progression [first line: HR, 0.97 (95% CI, 0.40-2.35); second line: HR, 0.91 (95% CI, 0.42-2.01); and third line: HR, 2.83 (95% CI, 0.73-11.03)]. By contrast, patients with non-TNBC appeared to improve with non-platinum-based chemotherapy [first line: HR, 2.57 (95% CI, 1.11-5.99); second line: HR, 1.91 (95% CI, 1.00-3.63); and third line: HR, 1.08 (95% CI, 0.53-2.18)]. Although the present study was limited by the sample size and its observational nature, the results indicated that platinum-based chemotherapy does not offer a discernible or distinct advantage compared with standard regimens in patients with TNBC, and is perhaps less efficacious in patients with non-TNBC.


Simplified Deployment of Health Informatics Applications by Providing Docker Images.
Due to the specific needs of biomedical researchers, in-house development of software is widespread. A common problem is to maintain and enhance software after the funded project has ended. Even if many tools are made open source, only a couple of projects manage to attract a user basis large enough to ensure sustainability. Reasons for this include complex installation and configuration of biomedical software as well as an ambiguous terminology of the features provided; all of which make evaluation of software laborious. Docker is a para-virtualization technology based on Linux containers that eases deployment of applications and facilitates evaluation. We investigated a suite of software developments funded by a large umbrella organization for networked medical research within the last 10 years and created Docker containers for a number of applications to support utilization and dissemination.


[Methods for early detection of predisposition to abnormal increases in body weight of children and adolescents].
This article presents the results of a special method designed for the early identification of children's and adolescents' propensity for abnormal weight gain. The basis of this technique is the principle of monitoring the actual values of body mass index for each child or young person at the age of dynamic changes in the index of the normal range. The weekly body weight of each child or adolescent who participates in the study is measured and the results are included in a special registry. Measurement of the body weight of the children and adolescents occurs in the morning at the same hour on an empty stomach after use of the toilet and performance of necessary hygiene. Afterwards, the obtained values of their body mass indices are compared with those considered normal for age, according to official data. In cases when the resulting body mass index of a child or young person exceeds the range of normal-for-age values, an in-depth medical examination of the child or adolescent is conducted together with the nature of their food intake and mode of physical activity, which may then be corrected.


Primary Microglia Isolation from Postnatal Mouse Brains.
Microglia are the mononuclear phagocytes in the central nervous system (CNS), which play key roles in maintaining homeostasis and regulating the inflammatory process in the CNS. To study the microglial biology in vitro, primary microglia show great advantages compared to immortalized microglial cell lines. However, microglia isolation from the postnatal mouse brain is relatively less efficient and time-consuming. In this protocol, we provide a quick and easy-to-follow method to isolate primary microglia from the neonatal mouse brain. The overall steps of this protocol include brain dissection, primary brain cell culture, and microglia isolation. Using this approach, researchers can obtain primary microglia with high purity. In addition, the harvested primary microglia were able to respond to the lipopolysaccharides challenge, indicating they retained their immune function. Collectively, we developed a simplified approach to efficiently isolate primary microglia with high purity, which facilitates a wide range of microglial biology investigations in vitro.


GNQ-209P Mutation in Metastatic Uveal Melanoma and Treatment Outcome.
Metastatic prognosis in uveal melanoma is assessed by gene expression profiling (GEP) testing of the tumor cells, usually obtained by fine needle aspiration (FNA). GEP has demonstrated high accuracy in distinguishing class I and II tumors, both having different metastatic potential. Transcriptomic studies identified distinct mutations including somatic mutations in GNAQ and GNA11, detected in more than 80%, and contribute to the upregulation of the mitogen-activated protein kinase (MAPK) pathway and the development of uveal melanoma (UM). The role of these mutations in treatment selection and possible benefit from targeted therapy are somewhat unclear. However, until the discovery of novel agents, local versus systemic therapies remain options for treatment that can still be considered for disease control in certain cases. We report a series of patients with metastatic UM with distinct mutational profiles. One had significant liver metastases with proven GNQ-209P mutation on tissue biopsy while peripheral blood molecular profiling did not show these mutations. The other three cases had no GNQ-209P mutation. All cases received nab-paclitaxel (Abraxane) as a treatment drug, and we record their responses to treatment and their molecular-profiling results.


Transcriptome analysis of a Pseudomonas aeruginosasn-glycerol-3-phosphate dehydrogenase mutant reveals a disruption in bioenergetics.
Pseudomonas aeruginosa causes acute and chronic human infections and is the major cause of morbidity and mortality in cystic fibrosis (CF) patients. We previously determined that the sn-glycerol-3-phosphate dehydrogenase encoded by glpD plays a larger role in P. aeruginosa physiology beyond its role in glycerol metabolism. To better understand the effect of a glpD mutation on P. aeruginosa physiology we compared the transcriptomes of P. aeruginosa strain PAO1 and the PAO1ΔglpD mutant using RNA-seq analysis. We determined that a null mutation of glpD significantly altered amino acid metabolism in P. aeruginosa and affected the production of intermediates that are channelled into the tricarboxylic acid cycle. Moreover, the loss of glpD induced a general stress response mediated by RpoS in P. aeruginosa. Several other phenotypes observed for the P. aeruginosa glpD mutant include increased persister cell formation, reduced extracellular ATP accumulation and increased heat output. Taken together, these findings implicate sn-glycerol-3-phosphate dehydrogenase as a key player in energy metabolism in P. aeruginosa.


Completeness and consistency of primary outcome reporting in COVID-19 publications in the early pandemic phase: a descriptive study.
The COVID-19 pandemic saw a steep increase in the number of rapidly published scientific studies, especially early in the pandemic. Some have suggested COVID-19 trial reporting is of lower quality than typical reports, but there is limited evidence for this in terms of primary outcome reporting. The objective of this study was to assess the prevalence of completely defined primary outcomes reported in registry entries, preprints, and journal articles, and to assess consistent primary outcome reporting between these sources. This is a descriptive study of a cohort of registered interventional clinical trials for the treatment and prevention of COVID-19, drawn from the DIssemination of REgistered COVID-19 Clinical Trials (DIRECCT) study dataset. The main outcomes are: 1) Prevalence of complete primary outcome reporting; 2) Prevalence of consistent primary outcome reporting between registry entry and preprint as well as registry entry and journal article pairs. We analyzed 87 trials with 116 corresponding publications (87 registry entries, 53 preprints and 63 journal articles). All primary outcomes were completely defined in 47/87 (54%) registry entries, 31/53 (58%) preprints and 44/63 (70%) journal articles. All primary outcomes were consistently reported in 13/53 (25%) registry-preprint pairs and 27/63 (43%) registry-journal article pairs. No primary outcome was specified in 13/53 (25%) preprints and 8/63 (13%) journal articles. In this sample, complete primary outcome reporting occurred more frequently in trials with vs. without involvement of pharmaceutical companies (76% vs. 45%), and in RCTs vs. other study designs (68% vs. 49%). The same pattern was observed for consistent primary outcome reporting (with vs. without pharma: 56% vs. 12%, RCT vs. other: 43% vs. 22%). In COVID-19 trials in the early phase of the pandemic, all primary outcomes were completely defined in 54%, 58%, and 70% of registry entries, preprints and journal articles, respectively. Only 25% of preprints and 43% of journal articles reported primary outcomes consistent with registry entries.


A study of inactivation of Chlamydia trachomatis by normal human serum.
To assess the effect of human serum on the viability of Chlamydia trachomatis, organisms were mixed with unheated and heat inactivated homologous serum, and the numbers surviving after incubation at 37 degrees C for 1 hour were compared. With a pool of sera obtained from 12 donors, the number of chlamydiae surviving incubation in unheated serum was less than 1% of that surviving incubation in heat inactivated serum. The antichlamydial activity of the unheated pooled serum samples could be noticeably reduced by treatment with Mg-EGTA (ethyleneglycolbis (beta-amino ethyl ether)-N,N'-tetra-acetic acid). This indicated a requirement for calcium ions and showed that the alternative pathway of complement activation played only a minor role, if any, in the inactivation process. When 12 serum samples were tested individually it was found that four inactivated chlamydiae to an extent comparable with that seen with the pooled serum. The other eight samples showed only moderate (or slight) antichlamydial activity, with survival rates in unheated serum of 20-60% (or more than 60%) of those in heat inactivated serum. There was no correlation between the titres of antichlamydial antibodies and antichlamydial activity, all serum samples having undetectable or low concentrations of antibody on measurement by micro-immunofluorescence. The antichlamydial activity destroyed by heating was restored, however, when heat inactivated serum was mixed with an equal volume of an unheated serum that was not inhibitory to chlamydiae. When the latter serum was heated before addition antichlamydial activity was not restored, indicating the requirement of both a heat stable and a heat labile factor. This observation and the need for calcium ions for inactivation of chlamydiae are compatable with killing mediated by antibody and complement. Thus serum samples from individuals with no clinical or serological evidence of infection with chlamydiae vary in their ability to inactivate the organism, some having antichlamydial activity which is possibly mediated by antibody and complement.


Tendon Injury and Fluoroquinolone Use: A Systematic Review.
Fluoroquinolone antibiotics are commonly used to treat infections and are prescribed by general practitioners, medical specialists and surgeons. Tendon injury has been associated with the use of these medications but the risk associated with newer fluoroquinolones has not been established. The aim of this systematic review was to evaluate the evidence from observational studies to determine the strength of the association between fluoroquinolone use and tendinopathy, and to identify risk factors for this complication. We searched MEDLINE, EMBASE and the Cochrane Collaboration from inception through May 2013 to identify observational studies focused on tendon injury and fluoroquinolones. Studies with original data were selected for inclusion following the PRISMA guidelines. Of the 560 abstracts screened, 16 relevant studies were independently rated by three authors (WW, AS, DC) using the Newcastle-Ottawa Quality Assessment Scale, and assigned a quality score out of 9. High-quality studies (i.e. scored 4.5 or higher) are summarized in detail in this article. Data were independently extracted by two authors (WW, AS). Overall, 16 studies were included in our study. Eight were deemed to be of high quality and five specifically evaluated Achilles tendon rupture. In addition, three studies examined Achilles tendinitis, and three included tendon disorders (including any tendon rupture) as an outcome. Results from these studies suggest that individuals exposed to fluoroquinolones are at increased risk for Achilles tendon rupture, particularly within the first month following exposure to the drug (odds ratios ranged from 1.1 to 7.1). One study showed an increased risk of tendon rupture in those over 60 years of age. Five studies stated that individuals taking fluoroquinolones and oral corticosteroids are at increased risk for tendon injury compared with those taking fluoroquinolones alone. Four studies examined the differential effect of a limited number of fluoroquinolones. Ofloxacin had the highest risk of tendon injury in three of the studies. Included studies are observational in nature and rely on self-report, which may lead to misclassification or underestimation of tendon injury. Observational studies showed an increased risk of tendon injury, including tendon rupture and tendinitis, with exposure to fluoroquinolone antibiotic therapy. Although this complication appears to be rare, concomitant corticosteroids increase the risk for tendon injury, which varies depending on the fluoroquinolone used.


Application of cryopreserved human hepatocytes in trichloroethylene risk assessment: relative disposition of chloral hydrate to trichloroacetate and trichloroethanol.
Trichloroethylene (TCE) is a suspected human carcinogen and a common groundwater contaminant. Chloral hydrate (CH) is the major metabolite of TCE formed in the liver by cytochrome P450 2E1. CH is metabolized to the hepatocarcinogen trichloroacetate (TCA) by aldehyde dehydrogenase (ALDH) and to the noncarcinogenic metabolite trichloroethanol (TCOH) by alcohol dehydrogenase (ADH). ALDH and ADH are polymorphic in humans, and these polymorphisms are known to affect the elimination of ethanol. It is therefore possible that polymorphisms in CH metabolism will yield subpopulations with greater than expected TCA formation with associated enhanced risk of liver tumors after TCE exposure. The present studies were undertaken to determine the feasibility of using commercially available, cryogenically preserved human hepatocytes to determine simultaneously the kinetics of CH metabolism and ALDH/ADH genotype. Thirteen human hepatocyte samples were examined. Linear reciprocal plots were obtained for 11 ADH and 12 ALDH determinations. There was large interindividual variation in the Vmax values for both TCOH and TCA formation. Within this limited sample size, no correlation with ADH/ALDH genotype was apparent. Despite the large variation in Vmax values among individuals, disposition of CH into the two competing pathways was relatively constant. These data support the use of cryopreserved human hepatocytes as an experimental system to generate metabolic and genomic information for incorporation into TCE cancer risk assessment models. The data are discussed with regard to cellular factors, other than genotype, that may contribute to the observed variability in metabolism of CH in human liver.


Two haplotype clusters of Echinococcus granulosus sensu stricto in northern Iraq (Kurdistan region) support the hypothesis of a parasite cradle in the Middle East.
Human cystic echinococcosis (CE) caused by Echinococcus granulosus s.s. is a major public health problem in Iraqi Kurdistan with a reported surgical incidence of 6.3 per 100,000 Arbil inhabitants. A total of 125 Echinococcus isolates retrieved from sheep, goats and cattle were used in this study. Our aim was to determine species/genotypes infecting livestock in Iraqi Kurdistan and examine intraspecific variation and population structure of Echinococcus granulosus s.s. in this region and relate it to that of other regions worldwide. Using nucleotide sequences of the mitochondrial cytochrome c oxidase subunit 1 (cox 1) we identified E. granulosus s.s. as the cause of hydatidosis in all examined animals. The haplotype network displayed a double-clustered topology with two main E. granulosus s.s. haplotypes, (KU05) and (KU33). The 'founder' haplotype (KU05) confirmed the presence of a common lineage of non-genetically differentiated populations as inferred by the low non-significant fixation index values. Overall diversity and neutrality indices indicated demographic expansion. We used E. granulosus s.s. nucleotide sequences from GenBank to draw haplotype networks for the Middle East (Iran, Jordan and Turkey), Europe (Albania, Greece, Italy, Romania and Spain), China, Mongolia, Russia, South America (Argentina, Brazil, Chile and Mexico) and Tunisia. Networks with two haplotype clusters like that reported here for Iraqi Kurdistan were seen for the Middle East, Europe, Mongolia, Russia and Tunisia using both 827bp and 1609bp cox1 nucleotide sequences, whereas a star-like network was observed for China and South America. We hypothesize that the double clustering seen at what is generally assumed to be the cradle of domestication may have emerged independently and dispersed from the Middle East to other regions and that haplotype (KU33) may be the main haplotype within a second cluster in the Middle East from where it has spread into Europe, Mongolia, Russia and North Africa. Further studies using metacestodes of human origin are required to investigate the biological importance of E. granulosus s.s. haplotypes/clusters and their association, if any with clinical manifestations of CE infection.


Anatomy of the Caucasian alar groove.
Anatomic studies performed on the noses of 15 cadavers examined the alar groove, alar lobule, and lower lateral crus areas both microscopically and on gross appearance to determine what effect these structures have on overall nasal appearance. In contrast to the findings of previous studies, the authors found the alar lobule to be an area in which dermis is interdigitated with muscle throughout and up to the alar rim. The anteroposterior lengths of the lower lateral crura were again seen to vary in length, presence or absence of accessory cartilages, and shape. Neither corrugation of the posterior elongation nor overlap of the accessory cartilages of the lower lateral cartilage had an effect on phenotype; sharp angles formed by the cartilage were blunted by the layer of fibrofatty muscular tissue between the cartilage and the skin. The alar groove, which lies at the junction of the lower lateral crus (medially) and the alar lobule (laterally), is defined not as much by a muscular attachment between the perichondrium of the lower lateral cartilage and the vestibular mucosa as by a bulging in the fatty layer on one side of the groove (within the cheek, lateral nasal wall, and nasal tip) and a relative paucity of fatty tissue on its other side (within the alar lobule).


An analysis of craniofacial form; the need for new analytic techniques.
This study was based on lateral cephalographs of 443 patients aged 7--18 years to determine the effects of age and occlusal (Angle) relationships on craniofacial form. From 33 datum points, 56 linear and angular dimensions were subjected to the multivariate statistical technique termed principal component analysis. This technique not only enabled a number of parameters to be combined together but also computed a series of orthogonal variates (components) accounting for over 90 percent of the total variance. The subsequent analysis of the component scores revealed marked contrasts in craniofacial form not only between age groups but also between different occlusal categories. The interpretation of such data remains controversial, since not only do traditional craniofacial parameters combine both size and shape together, but they are based upon datum points of unknown degrees of homology. This study therefore confirmed the complexity of craniofacial form and demonstrated the need to develop more appropriate analytic techniques with the specific objective of defining shape and shape change.


Characterization of the class 3 integron and the site-specific recombination system it determines.
Integrons capture gene cassettes by using a site-specific recombination mechanism. As only one class of integron and integron-determined site-specific recombination system has been studied in detail, the properties of a second class, the only known class 3 integron, were examined. The configuration of the three potentially definitive features of integrons, the intI3 gene, the adjacent attI3 recombination site, and the P(c) promoter that directs transcription of the cassettes, was similar to that found in the corresponding region (5' conserved segment) of class 1 integrons. The integron features are flanked by a copy of the terminal inverted repeat, IRi, from class 1 integrons on one side and a resolvase-encoding tniR gene on the other, suggesting that they are part of a transposable element related to Tn402 but with the integron module in the opposite orientation. The IntI3 integrase was active and able to recognize and recombine both known types of IntI-specific recombination sites, the attI3 site in the integron, and different cassette-associated 59-be (59-base element) sites. Both integration of circularized cassettes into the attI3 site and excision of integrated cassettes were also catalyzed by IntI3. The attI3 site was localized to a short region adjacent to the intI3 gene. Recombination between a 59-be and secondary sites was also catalyzed by IntI3, but at frequencies significantly lower than observed with IntI1, the class 1 integron integrase.


Moclobemide in the treatment of hot flashes in postmenopausal women.
This randomized, prospective, double-blind study evaluated the efficacy and tolerability of moclobemide, a reversible, selective inhibitor of monoamine oxidase-A, in reducing the frequency and severity of hot flashes. Thirty postmenopausal women were enrolled, and 28 were allocated to 5 weeks of treatment with moclobemide 150 mg (group 1, n = 10), moclobemide 300 mg (group 2, n = 11), or placebo (group 3, n = 9). Data on hot flashes were recorded in a daily diary. Mean reductions in the hot flash severity score were 24.4% in the placebo group, 69.8% in group 1, and 35.0% in group 2. This large difference suggests that the beneficial effects were not due to a placebo effect. Moclobemide may be a new nonhormonal option for reducing the incidence, severity, and duration of hot flashes in postmenopausal women who do not wish to take estrogen or have contraindications to its use.


Basic amino acid inhibition of cell division and macromolecular synthesis in Saccharomyces cerevisiae.
Growth of Saccharomyces cerevisiae on poor nitrogen sources such as allantoin or proline was totally inhibited by addition of a non-degradable basic amino acid to the medium. Cells treated with lysine contained greatly reduced quantities of histidine and arginine. Conversely, lysine and histidine were severely reduced in arginase-deficient cells treated with arginine. When all three basic amino acids were present in the culture medium, growth was normal suggesting that synthesis of all three basic amino acids was decreased by an excess of any one of them. Inhibition of growth was accompanied by a fivefold increase in the observed ratio of budded to unbudded cells. These morphological changes suggested that DNA synthesis was inhibited. Consistent with this suggestion, addition of a basic amino acid to the culture medium substantially reduced the ability of the cells to incorporate [14C]uracil into alkali-resistant, trichloroacetic acid-precipitable material. RNA and protein synthesis, although decreased, were less sensitive to the effects of such additions.


Optimal Care for Patients with Anaplastic Lymphoma Kinase (ALK)-Positive Non-Small Cell Lung Cancer: A Review on the Role and Utility of ALK Inhibitors.
The treatment of advanced non-small-cell lung cancer (NSCLC) has undergone a paradigm shift in the last decade. Molecular characterization of the disease has led to the rapid development of personalized medicine and swift delivery of targeted therapies to patients. The discovery of the anaplastic lymphoma kinase (ALK) gene in patients with NSCLC has resulted in rapid bench-bedside transition of several active drugs, with several others currently in clinical trials. After the first-generation ALK inhibitor crizotinib, next-generation ALK inhibitors have entered clinical applications for ALK-rearranged NSCLC. Ceritinib, alectinib, and brigatinib have all received approval for ALK-positive patients who have failed prior crizotinib, as well as first-line therapy in treatment-naïve patients based on favorable efficacy. Most recently, lorlatinib, a potent, newer-generation ALK inhibitor, has been approved as second- or third-line treatment. These advances have led to better patient outcomes, but concurrently have led to several crucial unanswered questions about optimal care for ALK-positive NSCLC patients. The ultimate acquisition of resistance to ALK-inhibitor therapy poses a challenge to ongoing research efforts, in addition to the routine management of these patients in the clinic. This review provides a summary of the clinical development of crizotinib, ceritinib, alectinib, brigatinib, and lorlatinib and highlights current management paradigms, current and evolving clinical information, emerging clinical decision-making and sequencing of therapy in advanced, metastatic, or recurrent ALK-positive NSCLC.


Mygalin, an Acanthoscurria gomesiana spider-derived synthetic, modulates haloperidol-induced cataleptic state in mice.
Haloperidol (HAL) is an antipsychotic used in the treatment of schizophrenia. However, adverse effects are observed in the extrapyramidal tracts due to its systemic action. Natural compounds are among the treatment alternatives widely available in Brazilian biodiversity. Mygalin (MY), a polyamine that was synthesized from a natural molecule present in the hemolymph of the Acanthoscurria gomesian spider, may present an interesting approach. This study aimed to evaluate the effect of MY in mice subjected to HAL-induced catalepsy. Male Swiss mice were used. Catalepsy was induced by intraperitoneal administration of HAL (0.5 mg/kg - 1 mL/Kg) diluted in physiological saline. To assess the MY effects on catalepsy, mice were assigned to 4 groups: (1) physiological saline (NaCl 0.9 %); (2) MY at 0.002 mg/Kg; (3) MY at 0.02 mg/Kg; (4) MY at 0.2 mg/Kg. MY or saline was administered intraperitoneally (IP) 10 min b HAL before saline. Catalepsy was evaluated using the bar test at 15, 30, 60, 90, and 120 min after the IP administration of HAL. The latency time in the bar test 15, 30, 60, and 90 min increased (p < 0.05) after IP administration of HAL compared to the control group. Catalepsy was attenuated 15, 30, 90, and 120 min (p < 0.05) after the IP-administration of MY at 0.2 mg/Kg; while MY at 0.02 mg/Kg attenuated catalepsy 15 min after the HAL treatment. Our findings showed that MY attenuates the HAL-induced cataleptic state in mice.


The Therapeutic Antibody LM609 Selectively Inhibits Ligand Binding to Human αVβ3 Integrin via Steric Hindrance.
The LM609 antibody specifically recognizes αVβ3 integrin and inhibits angiogenesis, bone resorption, and viral infections in an arginine-glycine-aspartate-independent manner. LM609 entered phase II clinical trials for the treatment of several cancers and was also used for αVβ3-targeted radioimmunotherapy. To elucidate the mechanisms of recognition and inhibition of αVβ3 integrin, we solved the structure of the LM609 antigen-binding fragment by X-ray crystallography and determined its binding affinity for αVβ3. Using single-particle electron microscopy, we show that LM609 binds at the interface between the β-propeller domain of the αV chain and the βI domain of the β3 chain, near the RGD-binding site, of all observed integrin conformational states. Integrating these data with fluorescence size-exclusion chromatography, we demonstrate that LM609 sterically hinders access of large ligands to the RGD-binding pocket, without obstructing it. This work provides a structural framework to expedite future efforts utilizing LM609 as a diagnostic or therapeutic tool.


Cytotoxicity of adenovirus-antibody aggregates: sensitivity to different cell strains, and inhibition by hexon antiserum and by complement.
Adenovirus-antibody aggregates under defined conditions are cytotoxic in vitro. All members of adenovirus groups I, II, and III caused toxicity upon aggregation. The toxicity of the clusters is exerted by the virions. Toxicity is temperature dependent and may be caused by a mechanism similar to that used in viral penetration. Cells permitting direct viral penetration were all sensitive to the toxic aggregates. The toxicity seems to be related to hexon antigens on the surface of the virions since antihexon sera neutralized the toxicity. No evidence was obtained showing that pentons are required for this kind of cytotoxicity. Adenovirus types 3, 5, and 9 were used in the experiment. Cytotoxicity was estimated by the (51)Cr release assay. Complement factors could be excluded as mediators of the cytolytic reactions. Instead, complement was shown to prevent the formation of toxic aggregates or to neutralize the toxicity of preformed ones.


Lymphocitic infundibulo-neurohypophysitis mimicking a pituitary adenoma.
A rare case of infundibulo-neurohypophysitis mimicking a pituitary adenoma is presented. A 69-years-old female patient developed polyuria and polydipsia. Laboratory analysis revealed central diabetes insipidus. No hormonal abnormalities. Cranial-magnetic resonance imaging (MRI) showed a left sided mass in the adenohypophysis presuming a pituitary adenoma. The mass had contact to both internal carotids. Admission to our department for neurosurgical treatment followed. Ophthalmologic examination and neurological examination yielded normal findings. A second MRI focussing on the sellar-region showed a left-sided (T2-MRI.hyperintense), distended adenohypophysis, without contrast enhancement in T1. The stalk appeared thickened. T1-weighted sequences of the neurohypophysis showed loss of signal intensity. We diagnosed an infundibulo-neurohypophysitis and abstained from surgical removal. The patient was discharged under treatment with corticosteroids and desmopressin. Hypophysitis is rare and shows special clinical characteristics. Despite defined radiological features to differentiate between hypophysitis and adenoma the possibility of misdiagnosis, and unnecessary surgical procedures, should always kept in mind.


Characterisation of crevicular fluid microbiota in primary Sjögren's syndrome.
To describe the taxonomy of the microbiota in crevicular fluid of primary Sjögren's syndrome (pSS) patients, and evaluate its association with clinical/serological variables, and oral quality of life. Observational study that included 48 pSS without diabetes mellitus, no active neoplasia, no antibiotic use in the previous two weeks, and no current active infection. We registered demographics, oral/ocular sicca symptoms, parotid enlargement and anti-Ro/La serology. We assessed the non-stimulated whole salivary flow (NSWSF), the EULAR Sjögren's Syndrome Patient Reported Index (ESSPRI), and the Xerostomia-related Quality of Life Scale (XeQoLS). Two periodontists determined the presence of periodontal disease and collected crevicular fluid from 6 teeth using filter paper. Samples were frozen at -86°C until processing. We included 17 sex- and age-matched control subjects. Bacterial DNA was extracted from the crevicular fluid sample using a commercial kit. 16SrRNA V3-V4 region was sequenced using reversible adaptor technology. Sequences were pre-processed and analysed using QIIME2 and phyloseq software programs. Functionality profiles were predicted using the Tax4Fun2 package. PSS patients had more bacteria of the genera Prevotella, Streptococcus, Veillonella, Fusobacterium, and Leptotrichia and fewer bacteria of the genus Selenomonas than controls. The pSS microbiota contained more genes encoding accessory secretory proteins. Microbiota also differed between patients with anti-Ro/La status, parotid gland enlargement, and periodontal disease severity, but did not correlate with NSWSF and XeQoLS. The crevicular fluid microbiota of pSS patients and controls differed significantly, even in SSP patients depending on their serology, parotid gland enlargement, and periodontal disease status.


Cytotoxicity and immunogenicity of SACCHACHITIN and its mechanism of action on skin wound healing.
SACCHACHITIN membrane, a weavable skin substitute made from the residual fruiting body of Ganoderma tsugae, has been demonstrated to promote skin wound healing. Prior to its clinical application, it is critical to learn more about any possible cytotoxicity, immunogenicity, or allergy response, and at least some of its mechanism(s) of action(s). In the present studies, it has been found that SACCHACHITIN suspension at less than 0.05% shows no cytotoxicity to the primary culture of rat fibroblasts. However, at higher concentrations (> or = 0.1%), it does reduce the growth of fibroblasts, based on MTT assays. This might be caused by positive charges on chitin molecules that are too strong, and may be harmful to the cell membrane. SACCHACHITIN showed no immunogenicity after it was inoculated into rats three times; however, the unmodified, purified rabbit type I and type II collagens did. Subcutaneous injection of SACCHACHITIN suspension into rats showed no gross allergic responses on skin. Nevertheless, it did cause local acute inflammation, as observed by histological investigation. This is similar to what occurred in the wound site covered with SACCHACHITIN membrane. The chemotactic effect of SACCHACHITIN was exhibited in both intact and wounded skin tissues. This may be one of the initial beneficial effects of SACCHACHITIN membrane to wound healing. The rapid acute inflammatory process was followed by the appearance of angiogenesis and granulation tissue formation, which occurred earlier than it normally would. Coverage of the wound area with SACCHACHITIN membrane also induced an earlier formation of scar tissue to replace the granulation tissue. A 1.5 x 1.5 cm(2) wound area covered by SACCHACHITIN completely healed by 21 days, while that covered with cotton gauze did not. Therefore, SACCHACHITIN is a safe biomaterial for use as a wound dressing for skin healing. Its promoting action for wound healing might be due to its chemotactic effect for inflammatory cells. This, in turn, may facilitate subsequent angiogenesis, granulation tissue formation, and faster new tissue formation, leading to faster wound healing.


Evidence for a replication function of FFA-1, the Xenopus orthologue of Werner syndrome protein.
DNA replication in higher eukaryotic cells occurs at a large number of discrete sites called replication foci. We have previously purified a protein, focus-forming activity 1 (FFA-1), which is involved in the assembly of putative prereplication foci in Xenopus egg extracts. FFA-1 is the orthologue of the Werner syndrome gene product (WRN), a member of the RecQ helicase family. In this paper we show that FFA-1 colocalizes with sites of DNA synthesis and the single-stranded DNA binding protein, replication protein A (RPA), in nuclei reconstituted in the egg extract. In addition, we show that two glutathione S-transferase FFA-1 fusion proteins can inhibit DNA replication in a dominant negative manner. The dominant negative effect correlates with the incorporation of the fusion proteins into replication foci to form "hybrid foci," which are unable to engage in DNA replication. At the biochemical level, RPA can interact with FFA-1 and specifically stimulates its DNA helicase activity. However, in the presence of the dominant negative mutant proteins, the stimulation is prevented. These results provide the first direct biochemical evidence of an important role for FFA-1 in DNA replication.


5,19-cyclo-9beta,10xi-androstane-3,17-dione promotes neurotrophic factor biosynthesis in 1321N1 human astrocytoma cells and improves passive avoidance learning impairment.
Since neurotrophic factors are essential for neurons to form neuronal networks and maintain neuronal functions, neurotrophic factor-like substances or inducers of neurotrophic factors can be useful for the treatment of serious neuronal diseases such as Alzheimer's and Parkinson's diseases. In the present study, we examined an effect of 5,19-cyclo-9beta,10xi-androstane-3,17-dione (CAD) on neurotrophic factor synthesis in glial cells and scopolamine-induced impairment of learning in mice. 1321N1 human astrocytoma cells promoted secretion of certain neurotrophic factors in response to CAD with no cytotoxicity, which caused dramatic neurite outgrowth in rat pheochromocytoma (PC12) cells. In fact, CAD significantly enhanced nerve growth factor (NGF) secretion and its gene expression in 1321N1 cells, in a time and concentration-dependent manner. Because second messengers such as cAMP, inositol 1,4,5-trisphosphates and Ca(2+) induce NGF gene expression, we measured activities of adenylyl cyclase and phospholipase C and intracellular Ca(2+) concentration in 1321N1 cells. However, CAD changed neither second messenger levels. CAD enhanced the gene expression of proto-oncogene, c-fos that is one of the components of transcription factor (AP-1). In addition to those above, the in vivo effects of CAD were also examined. Although injection of muscarinic receptor antagonist scopolamine impaired passive avoidance learning in mice, pretreatment with CAD significantly reversed the adverse effect in a dose-dependent manner. Taking these results together, CAD has enormous therapeutic potential for serious neuronal diseases.


Ecologically coherent population structure of uncultivated bacterioplankton.
Bacterioplankton are main drivers of biogeochemical cycles and important components of aquatic food webs. While sequencing-based studies have revealed how bacterioplankton communities are structured in time and space, relatively little is known about intraspecies diversity patterns and their ecological relevance. Here, we use the newly developed software POGENOM (POpulation GENomics from Metagenomes) to investigate genomic diversity and differentiation in metagenome-assembled genomes from the Baltic Sea, and investigate their genomic variation using metagenome data spanning a 1700 km transect and covering seasonal variation at one station. The majority of the investigated species, representing several major bacterioplankton clades, displayed population structures correlating significantly with environmental factors such as salinity and temperature. Population differentiation was more pronounced over spatial than temporal scales. We discovered genes that have undergone adaptation to different salinity regimes, potentially responsible for the populations' existence along with the salinity range. This in turn implies the broad existence of ecotypes that may remain undetected by rRNA gene sequencing. Our findings emphasize the importance of physiological barriers, and highlight the role of adaptive divergence as a structuring mechanism of bacterioplankton species.


Epidemiologic and Clinical Outcomes of Pediatric Renal Tumors in Korea: A Retrospective Analysis of The Korean Pediatric Hematology and Oncology Group (KPHOG) Data.
Renal tumors account for approximately 7% of all childhood cancers. These include Wilms tumor (WT), clear cell sarcoma of the kidney (CCSK), malignant rhabdoid tumor of the kidney (MRTK), renal cell carcinoma (RCC), congenital mesoblastic nephroma (CMN) and other rare tumors. We investigated the epidemiology of pediatric renal tumors in Korea. From January 2001 to December 2015, data of pediatric patients (0-18 years) newly-diagnosed with renal tumors at 26 hospitals were retrospectively analyzed. Among 439 patients (male, 240), the most common tumor was WT (n=342, 77.9%), followed by RCC (n=36, 8.2%), CCSK (n=24, 5.5%), MRTK (n=16, 3.6%), CMN (n=12, 2.7%), and others (n=9, 2.1%). Median age at diagnosis was 27.1 months (range 0-225.5) and median follow-up duration was 88.5 months (range 0-211.6). Overall, 32 patients died, of whom 17, 11, 1, and 3 died of relapse, progressive disease, second malignant neoplasm, and treatment-related mortality. Five-year overall survival and event free survival were 97.2% and 84.8% in WT, 90.6% and 82.1% in RCC, 81.1% and 63.6% in CCSK, 60.3% and 56.2% in MRTK, and 100% and 91.7% in CMN, respectively (p < 0.001). The pediatric renal tumor types in Korea are similar to those previously reported in other countries. WT accounted for a large proportion and survival was excellent. Non-Wilms renal tumors included a variety of tumors and showed inferior outcome, especially MRTK. Further efforts are necessary to optimize the treatment and analyze the genetic characteristics of pediatric renal tumors in Korea.


Cardiac CT Beyond Coronaries: Focus on Structural Heart Disease.
Cardiac computed tomography (CT) is an established non-invasive imaging tool for the assessment of coronary artery disease. Furthermore, it plays a key role in the preinterventional work-up of patients presenting with structural heart disease. CT is the gold standard for preprocedural annular assessment, device sizing, risk determination of annular injury, coronary occlusion or left ventricular outflow tract obstruction, calcification visualization and quantification of the target structure, and prediction of a co-planar fluoroscopic angulation for transcatheter interventions in patients with structural heart disease. It is further a key imaging modality in postprocedural assessment for prosthesis thrombosis, degeneration, or endocarditis. CT plays an integral part in the imaging work-up of novel transcatheter therapies for structural heart disease and postprocedural assessment for prosthesis thrombosis or endocarditis. This review provides a comprehensive overview of the key role of CT in the context of structural heart interventions.


Activation of liver X receptor decreases atherosclerosis in Ldlr⁻/⁻ mice in the absence of ATP-binding cassette transporters A1 and G1 in myeloid cells.
Liver X receptor (LXR) activators decrease atherosclerosis in mice. LXR activators (1) directly upregulate genes involved in reverse cholesterol transport and (2) exert anti-inflammatory effects mediated by transrepression of nuclear factor-κB target genes. We investigated whether myeloid cell deficiency of ATP-binding cassette transporters A1 and G1 (ABCA1/G1), principal targets of LXR that promote macrophage cholesterol efflux and initiate reverse cholesterol transport, would abolish the beneficial effects of LXR activation on atherosclerosis. LXR activator T0901317 substantially reduced inflammatory gene expression in macrophages lacking ABCA1/G1. Ldlr(-/-) mice were transplanted with Abca1(-/-)Abcg1(-/-) or wild-type bone marrow (BM) and fed a Western-type diet for 6 weeks with or without T0901317 supplementation. Abca1/g1 BM deficiency increased atherosclerotic lesion complexity and inflammatory cell infiltration into the adventitia and myocardium. T0901317 markedly decreased lesion area, complexity, and inflammatory cell infiltration in the Abca1(-/-)Abcg1(-/-) BM-transplanted mice. To investigate whether this was because of macrophage Abca1/g1 deficiency, Ldlr(-/-) mice were transplanted with LysmCreAbca1(fl/fl)Abcg1(fl/fl) or Abca1(fl/fl)Abcg1(fl/fl) BM and fed Western-type diet with or without the more specific LXR agonist GW3965 for 12 weeks. GW3965 decreased lesion size in both groups, and the decrease was more prominent in the LysmCreAbca1(fl/fl)Abcg1(fl/fl) group. The results suggest that anti-inflammatory effects of LXR activators are of key importance to their antiatherosclerotic effects in vivo independent of cholesterol efflux pathways mediated by macrophage ABCA1/G1. This has implications for the development of LXR activators that lack adverse effects on lipogenic genes while maintaining the ability to transrepress inflammatory genes.


Do-it-yourself strategic planning provides map to the future.
An alternative to spending a six-figure sum for a consultant-produced strategic plan may be for the staff of a healthcare organization to develop a strategic plan as a "do-it-yourself" project. In addition to being less expensive to acquire, such a plan offers staff the opportunity to control the plan's content tightly and thus make it easier to implement.


Remote triage incorporating symptom-based risk stratification for suspected head and neck cancer referrals: A prospective population-based study.
Remote triage for suspected head and neck cancer (HNC) referrals was adopted by many institutions during the initial peak of the coronavirus disease 2019 pandemic. Its safety in this population has not been established. A 16-week, prospective, multicenter national service evaluation was started on March 23, 2020. Suspected HNC referrals undergoing remote triage in UK secondary care centers were identified and followed up for a minimum of 6 months to record the cancer status. Triage was supported by risk stratification using a validated calculator. Data for 4568 cases were submitted by 41 centers serving a population of approximately 26 million. These represented 14.1% of the predicted maximum referrals for this population outside of pandemic times, and this gave the study a margin of error of 1.34% at 95% confidence. Completed 6-month follow-up data were available for 99.8% with an overall cancer rate of 5.6% (254 of 4557). The rates of triage were as follows: urgent imaging investigation, 25.4% (n = 1156); urgent face-to-face review, 27.8%; (n = 1268); assessment deferral, 30.3% (n = 1382); and discharge, 16.4% (n = 749). The corresponding missed cancers rates were 0.5% (5 of 1048), 0.3% (3 of 1149), 0.9% (12 of 1382), and 0.9% (7 of 747; P = .15). The negative predictive value for a nonurgent triage outcome and no cancer diagnosis was 99.1%. Overall harm was reported in 0.24% (11 of 4557) and was highest for deferred assessments (0.58%; 8 of 1382). Remote triage, incorporating risk stratification, may facilitate targeted investigations for higher risk patients and prevent unnecessary hospital attendance for lower risk patients. The risk of harm is low and may be reduced further with appropriate safety netting of deferred appointments. This large national study observed the widespread adoption of telephone assessment (supported by a risk calculator) of patients referred to hospital specialists with suspected head and neck cancer during the initial peak of the coronavirus disease 2019 pandemic. The authors identified 4568 patients from 41 UK centers (serving a population of more than 26 million people) who were followed up for a minimum of 6 months. Late cancers were identified, whether reviewed or investigated urgently (0.4%) or nonurgently (0.9%), but the overall rate of harm was low (0.2%), with the highest rate being seen with deferred appointments (0.6%).


Acute cholecystitis. Evaluation of factors influencing common duct exploration.
The clinical records of 216 patients with proven acute cholecystitis treated by cholecystectomy form the basis of this retrospective study. Common bile duct stones were present in 12.4 per cent of these patients. Thirty per cent of the patients with elevated SGOT values, 26.2 per cent of the patients with elevated alkaline phosphatase, and 23.1 per cent of the patients with elevated amylase had common duct stones. The authors found that 17.6 per cent of patients with bilirubin between 1.5 and 2.9 mg/dl had common duct stones, whereas 71.4 per cent of common bile ducts with a bilirubin greater than 5 mg/dl contained stones. Six of 28 patients with common duct stones had normal bilirubin. Cholangiograms were normal in 115 of the 154 cholangiograms performed; six of these common ducts were explored, and no common duct stones found (false-negative cholangiograms 0.0%). Cholangiograms showed stones in 24 patients; common bile duct stones were recovered from 20 of these patients (accuracy rate 83%, false-positive cholangiograms 17%). Wound infections occurred in seven patients (3.7%). Sepsis resulted in death of three patients, and the other two deaths resulted from multi-system failure. This study demonstrates operative cholangiograms to be the most accurate method of detection of common duct stones, and its routine use in patients undergoing cholecystectomy is recommended.


Current approaches to neurobehavioural testing in occupational health.
In recent years neurobehavioural tests have been used increasingly in occupational settings to identify changes in cognitive function associated with exposure to neurotoxicants. Potential applications of these methods in occupational health include research, diagnosis, and screening. Applications in cross sectional research studies, involving the comparison of the performance of exposed and control groups, are well established. However, the use of such methods requires attention to factors other than exposure which may influence test performance. The validity of adapting existing test batteries for diagnosis or screening is questionable. Well developed techniques exist for diagnosis but this requires lengthy and skilled test administration and interpretation and cannot be accomplished with research batteries. The application of neurobehavioural methods for screening currently presents several difficulties. Current problems and future directions in this field are discussed.


Synthesis of deoxyribonucleic acid in BHK-21-C13 cells.
Newly synthesized (3)H-labelled DNA was extracted from baby hamster kidney cells (BHK-21/C13 cells) and was shown to possess single-stranded properties when examined by column chromatography on benzoylated naphthoylated DEAE-cellulose, hydroxyapatite and methylated albumin on kieselguhr, and by its affinity for nitrocellulose filters. Some of the newly synthesized DNA was shown to be of lower molecular weight than the bulk of the DNA when examined by alkaline sucrose-density-gradient centrifugation. The properties observed were not affected by treatment of the DNA with ribonuclease, Pronase or amylase. The effect of the size of the DNA on its observed properties was examined and is discussed. It is concluded that DNA synthesis in BHK-21/C13 cells proceeds according to the discontinuous-mechanism model in at least one of the strands.


NEK2 enhances malignancies of glioblastoma via NIK/NF-κB pathway.
Glioblastoma (GBM) is one of the most lethal primary brain tumor with a poor median survival less than 15 months. Despite the development of the clinical strategies over the decades, the outcomes for GBM patients remain dismal due to the strong proliferation and invasion ability and the acquired resistance to radiotherapy and chemotherapy. Therefore, developing new biomarkers and therapeutic strategies targeting GBM is in urgent need. In this study, gene expression datasets and relevant clinical information were extracted from public cancers/glioma datasets, including TCGA, GRAVENDEEL, REMBRANDT, and GILL datasets. Differentially expressed genes were analyzed and NEK2 was picked as a candidate gene for subsequent validation. Human tissue samples and corresponding data were collected from our center and detected by immunohistochemistry analysis. Molecular biological assays and in vivo xenograft transplantation were performed to confirm the bioinformatic findings. High-throughput RNA sequencing, followed by KEGG analysis, GSEA analysis and GO analysis were conducted to identify potential signaling pathways related to NEK2 expression. Subsequent mechanism assays were used to verify the relationship between NEK2 and NF-κB signaling. Overall, we identified that NEK2 is significantly upregulated in GBM and the higher expression of NEK2 exhibited a poorer prognosis. Functionally, NEK2 knockdown attenuated cell proliferation, migration, invasion, and tumorigenesis of GBM while NEK2 overexpression promoted the GBM progression. Furthermore, High-throughput RNA sequencing and bioinformatics analysis indicated that NEK2 was positively related to the NF-κB signaling pathway in GBM. Mechanically, NEK2 activated the noncanonical NF-κB signaling pathway by phosphorylating NIK and increasing the activity and stability of NIK. In conclusion, NEK2 promoted the progression of GBM through activation of noncanonical NF-κB signaling, indicating that NEK2- NF-κB axis could be a potential drug target for GBM.


BLINDNESS RELATED TO PRESUMED RETINAL TOXICITY AFTER USING PERFLUOROCARBON LIQUID DURING VITREORETINAL SURGERY.
To describe the presumed retinal toxicity after using specific batches of perfluorocarbon liquid ALA OCTA (Alamedics, Dornstadt, Germany) in pars plana vitrectomy. This is an observational retrospective consecutive case series analyses of patients operated on pars plana vitrectomy for retinal detachment or intraocular lens subluxation, using the 150141 or 200114 batches of perfluorocarbon liquid ALA OCTA as assistance during the surgery in a single center. Patients were included in this report if they manifested retinal toxicity signs throughout the follow-up, such as retinal and retinal pigment epithelium atrophy, disk paleness, and intensive macular fibrosis. Spectral domain optical coherence tomography (Spectralis; Heidelberg Engineering, Heidelberg, Germany) and Ultra-Wide Field 200° retinal camera (Optos P200Tx; Optos, Scotland, United Kingdom) images, electrophysiological tests, and visual fields were performed to analyze the retinal structure and functionality. Seven of 80 patients showed all the described signs of toxicity, after a mean follow-up of 34.29 days (range: 10-87) since surgery. Four patients needed a second pars plana vitrectomy because of tractional retinal detachment and proliferative vitreoretinopathy, and two of them underwent a third surgery because of redetachment. All patients experienced amaurosis or central scotoma, with a final best-corrected visual acuity ranging from 20/200 to light perception. Presumed toxic batches of perfluorocarbon liquid may cause massive retinal toxicity. A rapid suspicion, a correct traceability of surgical products, and informing health authorities are fundamental to prevent further cases of toxicity.


[Comparative assessment of efficacy of local thrombolysis with various fibrinolytics].
To avoid the characteristic for thrombolytic therapy of pulmonary embolism problems associated with low efficacy and elevated danger haemorrhagic complications may, in our opinion, be possible by means of the method of local thrombolysis worked out in our Clinic (RF Patent No 2376042). The present work was aimed at assessing efficacy of local thrombolysis in patients diagnosed as having pulmonary artery embolism with the drugs urokinase and actilyse. We examined a total of twenty patients. Ten Group One patients underwent local thrombolysis with Urokinase, and the remaining ten Group Two patients were treated with actilyse. We studied the parameters of central and pulmonary haemodynamics. Local thrombolysis with urokinase and actilyse in patients with pulmonary artery embolism turned out to equally efficiently reestablish blood flow in the pulmonary artery, which was confirmed by normalization of central haemodynamics maximally 2 hours after carrying out thrombolytic therapy with urokinase, with acktilyse accelerating this process to take 1 hour only. The proposed method makes it possible to decrease the dose of fibrinolytic agents twofold as compared with the recommended guidelines of the Pharmacopoeia, without losing their efficacy and ensures prevention of haemorrhagic complications.


The Cambia Sojourns Scholars Leadership Program: Conversations with Emerging Leaders in Palliative Care.
There is a pressing workforce shortage and leadership scarcity in palliative care to adequately meet the demands of individuals with serious illness and their families. To address this gap, the Cambia Health Foundation launched its Sojourns Scholars Leadership Program in 2014, an initiative designed to identify, cultivate, and advance the next generation of palliative care leaders. This report intends to summarize the second cohort of Sojourns Scholars' projects and their reflection on their leadership needs. This report summarizes the second cohort of sojourns scholars' project and their reflection on leadership needs. After providing a written reflection on their own projects, the second cohort participated in a group interview (fireside chat) to elicit their perspectives on barriers and facilitators in providing palliative care, issues facing leadership in palliative care in the United States, and lessons from personal and professional growth as leaders in palliative care. They analyzed the transcript of the group interview using qualitative content analysis methodology. Three themes emerged from descriptions of the scholars' project experience: challenges in palliative care practice, leadership strategies in palliative care, and three lessons learned to be a leader were identified. Challenges included perceptions of palliative care, payment and policy, and workforce development. Educating and collaborating with other clinicians and influencing policy change are important strategies used to advance palliative care. Time management, leading team effort, and inspiring others are important skills that promote effectiveness as a leader. Emerging leaders have a unique view of conceptualizing contemporary palliative care and shaping the future. Providing comprehensive, coordinated care that is high quality, patient and family centered, and readily available depends on strong leadership in palliative care. The Cambia Scholars Program represents a unique opportunity.


Expression of claudin-7 in benign kidney and kidney tumors.
Claudins, a family of tight junction-related transmembrane proteins, have been implicated in the pathogenesis of various human neoplasms. Expression of claudin-7 was increased in chromophobe renal cell carcinoma in a recent oligonucleotide microarray study. We studied the expression of claudin-7 in benign and neoplastic kidneys by immunohistochemical staining. Distal nephron (distal convoluted tubule and thick ascending limb of Henle's loop) epithelium showed strong membranous staining in 100% (174/174) of the cases. Chromophobe renal cell carcinoma was positive for claudin-7 expression in 100% (36/36) of cases, while papillary renal cell carcinoma, oncocytoma and clear cell renal cell carcinoma were positive in 90% (71/80), 45% (21/47) and 7% (7/98) of the cases, respectively. Differential expression of Claudin-7 in different types of renal cell neoplasms can be useful in their differential diagnosis, particularly when used in a panel of markers. In addition, results from this study support previous reports of distal nephron origin for chromophobe renal cell carcinoma and oncocytoma. The data also suggest that, as far as claudin-7 expression is concerned, papillary renal cell carcinoma may be more closely related to the distal nephron, rather than the proximal nephron.


Effects of consecutive trials and test-retest reliability of 1000-Hz tympanometry in adults.
To investigate the effect of consecutive 1000-Hz tympanometry testing on admittance measures, and examine test-retest reliability. Repeated measures with eight trials of 1000-Hz Y(a) and B(a)/G(a) tympanometry, respectively, in two ears of each subject, followed by repeated 226-Hz tympanometry. Twenty-seven normal-hearing young adults. For single-peak tympanograms, peak Y(tm) and G(tm) systematically increased across trials with a mean change of 8% and 15% at Trial 8, respectively, whereas B(tm) increased marginally. For notched tympanograms, peak Y(tm) and B(tm) decreased by 23% and 162% at Trial 8, and G(tm) in two cases also decreased (10% on average). Trial 2 and 3 contributed 50% to 70% of the total changes. Test-retest differences of subsequently acquired 1000- and 226-Hz tympanograms were smaller than previously reported. Consecutive testing significantly alters middle-ear admittance in 1000-Hz tympanometry. The outcome is contingent on tympanogram pattern and admittance component: Increase of peak Y(tm) and G(tm) in single-peak tympanograms and decrease of all measures in notched tympanograms. The present results complement previous studies on our understanding of the mechanism underlying this effect: a decrease of middle-ear stiffness. The effect of repetitive tympanometry should be accounted for in research involving sequential testing.


Population pharmacokinetics of topiramate in adult patients with epilepsy using nonlinear mixed effects modelling.
The objective of the study was to develop population pharmacokinetic model of topiramate (TPM) using nonlinear mixed effects modelling approach. Data were collected from 78 adult epileptic patients on mono- or co-therapy of TPM and other antiepileptic drugs, such as carbamazepine (CBZ), valproic acid, lamotrigine, levetiracetam, phenobarbital and pregabalin. Steady-state TPM concentrations were determined in blood samples by high performance liquid chromatography with fluorescence detection. A one-compartment model with first order absorption and elimination was used to fit the concentration-time TPM data. Volume of distribution of TPM was estimated at 0.575 l/kg. The influence of demographic, biochemical parameters and therapy characteristics of the patients on oral clearance (CL/F) was evaluated. Daily carbamazepine dose (DCBZ) and renal function estimated by Modification of diet in renal disease (MDRD) formula significantly (p<0.001) influenced CL/F and were included in the final model: CL/F · (l/h)=1.53(l/h) · [1+0.476 · DCBZ(mg/day)/1000(mg/day)] · EXP[0.00476 · [MDRD(ml/ min)-95.72(ml/min)]]. Increase of CL/F with DCBZ and MDRD was best described by linear and exponential models. Mean TPM CL/F during CBZ co-therapy was 2.46 l/h, which is higher for 60.8% than in patients not co-treated with CBZ. Evaluation by bootstrapping showed that the final model was stable. The predictive performance was evaluated by adequate plots and indicated satisfactory precision. This model allows individualisation of TPM dosing in routine patient care, especially useful for patients on different CBZ dosing regimen.


CTCF-mediated 3D chromatin predetermines the gene expression program in the male germline.
Spermatogenesis is a unidirectional differentiation process that generates haploid sperm, but how the gene expression program that directs this process is established is largely unknown. Here we determine the high-resolution 3D chromatin architecture of male germ cells during spermatogenesis and show that CTCF-mediated 3D chromatin predetermines the gene expression program required for spermatogenesis. In undifferentiated spermatogonia, CTCF-mediated chromatin contacts on autosomes pre-establish meiosis-specific super-enhancers (SE). These meiotic SE recruit the master transcription factor A-MYB in meiotic spermatocytes, which strengthens their 3D contacts and instructs a burst of meiotic gene expression. We also find that at the mitosis-to-meiosis transition, the germline-specific Polycomb protein SCML2 resolves chromatin loops that are specific to mitotic spermatogonia. Moreover, SCML2 and A-MYB establish the unique 3D chromatin organization of sex chromosomes during meiotic sex chromosome inactivation. We propose that CTCF-mediated 3D chromatin organization enforces epigenetic priming that directs unidirectional differentiation, thereby determining the cellular identity of the male germline.


Temporal evolution of the microbiome, immune system and epigenome with disease progression in ALS mice.
Amyotrophic lateral sclerosis (ALS) is a terminal neurodegenerative disease. Genetic predisposition, epigenetic changes, aging and accumulated life-long environmental exposures are known ALS risk factors. The complex and dynamic interplay between these pathological influences plays a role in disease onset and progression. Recently, the gut microbiome has also been implicated in ALS development. In addition, immune cell populations are differentially expanded and activated in ALS compared to healthy individuals. However, the temporal evolution of both the intestinal flora and the immune system relative to symptom onset in ALS is presently not fully understood. To better elucidate the timeline of the various potential pathological factors, we performed a longitudinal study to simultaneously assess the gut microbiome, immunophenotype and changes in ileum and brain epigenetic marks relative to motor behavior and muscle atrophy in the mutant superoxide dismutase 1 (SOD1G93A) familial ALS mouse model. We identified alterations in the gut microbial environment early in the life of SOD1G93A animals followed by motor dysfunction and muscle atrophy, and immune cell expansion and activation, particularly in the spinal cord. Global brain cytosine hydroxymethylation was also altered in SOD1G93A animals at disease end-stage compared to control mice. Correlation analysis confirmed interrelationships with the microbiome and immune system. This study serves as a starting point to more deeply comprehend the influence of gut microorganisms and the immune system on ALS onset and progression. Greater insight may help pinpoint novel biomarkers and therapeutic interventions to improve diagnosis and treatment for ALS patients.This article has an associated First Person interview with the joint first authors of the paper.


Utilization of a Peritoneal Interposition Flap to Prevent Symptomatic Lymphoceles After Robotic Radical Prostatectomy and Bilateral Pelvic Lymph Node Dissection.
Introduction: The peritoneal interposition flap (PIF) has been shown to prevent postoperative symptomatic lymphocele (SL) formation after robot-assisted radical prostatectomy (RARP) and pelvic lymph node dissection (PLND). The PIF inhibits the mobilized bladder from resealing over its lateral dissection planes, which overly the lymphadenectomy beds. This creates a window for lymphatic fluid to drain into the peritoneal cavity where it can be absorbed. Herein, we externally validate its utility in preventing postoperative SL formation and assess its effect on postoperative urinary function. Materials and Methods: We retrospectively reviewed all consecutive patients who underwent RARP with bilateral PLND by a single surgeon between July 2016 and September 2019. All patients who underwent surgery before August 8, 2018 did not receive the PIF, while those who underwent surgery after August 8, 2018 received the PIF. Our PIF technique involves fixing the peritoneum overlying the lateral dome of the bladder to the ipsilateral, anterior-lateral surface of the bladder using a barbed absorbable suture. Continuous and categorical variables were compared between the two groups using independent t-tests and chi-square tests, respectively; p < 0.05 was considered significant. Results: Of 318 total patients, 201 did not undergo the PIF and 117 underwent the PIF. With regard to postoperative complications, patients undergoing the PIF had a lower incidence of SL compared with those not undergoing the PIF (0.0% vs 6.0%, p = 0.007). There was no difference in 30-day postoperative nonlymphocele complications (Clavien >2) between both groups (p = 0.800). With regard to urinary function, there was no difference in the rate of 3-month postoperative continence (p = 0.624), preoperative American Urological Association Symptom Score (AUASS) (p = 0.898), and postoperative AUASS (p = 0.470) between both groups. Conclusion: Utilization of a PIF may minimize the risk of SL formation after RARP and PLND without increasing the risk of non-SL-related complications. This technique does not adversely affect postoperative urinary function.


Gated molecular baskets.
In this review, we describe the construction of gated molecular baskets, discuss their mechanism of action in regulating the exchange of guests and illustrate the potential of these concave hosts to act as catalysts for controlling chemical reactions. Importantly, a number of computational and experimental studies have suggested that gated baskets ought to unfold their gates at the rim for permitting the passage of guests to/from their inner space. These dynamic hosts are therefore offered as useful models for investigating the process of gating in artificial systems. Furthermore, gated baskets should permit examining the benefit of controlling the rate by which reactants access a gated catalyst for promoting chemical reactions occurring in its confined space.


Profound biotinidase deficiency in two asymptomatic adults.
Biotinidase deficiency is an autosomal-recessive disorder of biotin recycling. Children with profound biotinidase deficiency usually have neurological and cutaneous symptoms in early childhood, but they may not develop symptoms until adolescence. We now report on a man and a woman with profound biotinidase deficiency who are asymptomatic and who were diagnosed only because their biotinidase-deficient children were identified by newborn screening. These adults have never exhibited symptoms of the disorder and are homozygous for two different mutations resulting in different aberrant enzymes. There is no evidence of an increased dietary intake of biotin to explain why they have remained asymptomatic. Although these adults may still be at risk for developing symptoms, they could represent a small group of individuals with profound biotinidase deficiency who will never develop clinical problems. Their lack of symptoms suggests that there are probably epigenetic factors that protect some enzyme-deficient individuals from developing symptoms. These individuals broaden the spectrum of expression of biotinidase deficiency.