[Substrate specificity and action mechanism of glycosidases].
Result of author's research and data from literature have been generalized with respect to hydrolase and transferase activity of glycosidases: alpha-galactosidase and alpha-N-acetylgalactosaminidase--the enzymes which catalyse hydrolysis of natural and synthetic glycosides. Broad variability of action specificity of glycosidases with respect to glycon, aglycon as well as the bond type depending on the enzyme isolation source have been shown. One can suppose that the enzyme action specificity is connected with different formation mechanisms of enzyme-substrate complexes. An idea is discussed concerning the identity of the mechanism of splitting of various glycosidic links by the studied enzymes.


Teaching patient-centered communication skills: a telephone follow-up curriculum for medical students.
To encourage medical students' use of patient-centered skills in core clerkships, we implemented and evaluated a Telephone Follow-up Curriculum focusing on three communication behaviors: tailoring education to patients' level of understanding, promoting adherence by anticipating obstacles, and ensuring comprehension by having patients repeat the plans. The intervention group consisted of two different cohorts of third-year medical students in longitudinal clerkships (n=41); traditional clerkship students comprised the comparison group (n = 185). Intervention students telephoned one to four patients 1 week after seeing them in outpatient clinics or inpatient care to follow up on recommendations. We used surveys, focus groups, and clinical performance examinations to assess student perception, knowledge and skills, and behavior change. Students found that the curriculum had a positive impact on patient care, although some found the number of calls excessive. Students and faculty reported improvement in students' understanding of patients' health behaviors, knowledge of patient education, and attitudes toward telephone follow-up. Few students changed patient education behaviors or called additional patients. Intervention students scored higher in some communication skills on objective assessments. A patient-centered communication curriculum can improve student knowledge and skills. While some intervention students perceived that they made too many calls, our data suggest that more calls, an increased sense of patient ownership, and role modeling by clerkship faculty may ensure incorporation and application of skills.


Implementing guidelines for cancer pain management: results of a randomized controlled clinical trial.
Pain and symptom management is an integral part of the clinical practice of oncology. A number of guidelines have been developed to assist the clinician in optimizing comfort care. We implemented clinical guidelines for cancer pain management in the community setting and evaluated whether these guidelines improved care. Eighty-one cancer patients, aged 37 to 76 years, were enrolled onto a prospective, longitudinal, randomized controlled study from the outpatient clinic settings of 26 western Washington-area medical oncologists. A multilevel treatment algorithm based on the Agency for Health Care Policy and Research Guidelines for Cancer Pain Management was compared with standard-practice (control) pain and symptom management therapies used by community oncologists. The primary outcome of interest was pain (Brief Pain Inventory); secondary outcomes of interest were all other symptoms (Memorial Symptom Assessment Scale) and quality of life (Functional Assessment of Cancer Therapy Scale). Patients randomized to the pain algorithm group achieved a statistically significant reduction in usual pain intensity, measured as slope scores, when compared with standard community practice (P < .02). Concurrent chemotherapy and patient adherence to treatment were significant mediators of worst pain. There were no significant differences in other symptoms or quality of life between the two treatment groups. This guideline implementation study supports the use of algorithmic decision making in the management of cancer pain. These findings suggest that comprehensive pain assessment and evidence-based analgesic decision-making processes do enhance usual pain outcomes.


A GIS based method for soil mapping in Sardinia, Italy: a geomatic approach.
A new project was recently initiated for the realization of the "Land Unit and Soil Capability Map of Sardinia" at a scale of 1:50,000 to support land use planning. In this study, we outline the general structure of the project and the methods used in the activities that have been thus far conducted. A GIS approach was used. We used the soil-landscape paradigm for the prediction of soil classes and their spatial distribution or the prediction of soil properties based on landscape features. The work is divided into two main phases. In the first phase, the available digital data on land cover, geology and topography were processed and classified according to their influence on weathering processes and soil properties. The methods used in the interpretation are based on consolidated and generalized knowledge about the influence of geology, topography and land cover on soil properties. The existing soil data (areal and point data) were collected, reviewed, validated and standardized according to international and national guidelines. Point data considered to be usable were input into a specific database created for the project. Using expert interpretation, all digital data were merged to produce a first draft of the Land Unit Map. During the second phase, this map will be implemented with the existing soil data and verified in the field if also needed with new soil data collection, and the final Land Unit Map will be produced. The Land Unit and Soil Capability Map will be produced by classifying the land units using a reference matching table of land capability classes created for this project.


Overwintering of Heliothis virescens (F.) and Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) in cotton fields of northeast Mississippi.
Following the 1995-2001 crop seasons, population densities of pupae of tobacco budworm, Heliothis virescens (F.), and corn earworm, Helicoverpa zea (Boddie), overwintering in cotton fields, Gossypium hirsutum L., in Monroe County, MS, were estimated by digging a total of 43,542 m-row. Densities of pupae varied greatly among years with the highest densities occurring in years with low rainfall in the late summer and early fall. The sex ratio of pupae was variable and significantly male-biased in some years. The total area-wide, tobacco budworm population emerging from all overwintering sites was estimated from catches in pheromone traps whose absolute efficiency is known. After the introduction of Bacillus thuringiensis Berliner (Bt)-transgenic cotton in 1996 but before a several-fold increase in use of herbicide resistant (HR)-transgenic cotton cultivars in 2001, local cotton fields accounted for <2% of the total overwintered tobacco budworm population. In the pretransgenic cotton production era, it is estimated cotton fields typically accounted for <10% of the overwintered tobacco budworm population. Driving surveys of the tillage status of cotton fields and pheromone trapping to determine the timing of adult emergence in the spring indicate that--until the 2000 crop season--postseason tillage destroyed most tobacco budworm and corn earworm pupae overwintering in cotton fields. Adoption of HR-transgenic cultivars has quintupled cotton field area planted no-till, but the contribution of cotton fields to overwintered tobacco budworm populations has remained low. Preemptive tillage of cotton fields to kill overwintering tobacco budworm pupae would not appreciably suppress in-season population densities of this pest. However, the absence of such tillage might increase the rate at which the tobacco budworm develops resistance to insecticides and counteradaptation to antibiotic host plant resistance traits including the Bt-transgenic trait.


HeLaTG cells have mitochondrial DNA inserted into the c-myc oncogene.
To determine whether mitochondrial DNA (mtDNA) fragments found within the nucleus are transcribed, we have differentially screened a HeLaTG cDNA library. A clone that hybridized to mtDNA as well as to c-myc was identified. Analysis of the cDNA disclosed that it contained a mtDNA sequence, encoding cytochrome-c oxidase subunit III (coxIII) that was contiguous with and 5' of a c-myc sequence corresponding to part of exon 2 and exon 3. Hybridization of ScaI-digested DNA with a 1.05 kb c-myc probe revealed a unique band in HeLaTG cells, as well as a band common to HeLaTG and 13 other cell types examined. Solution hybridization of HeLaTG RNA with a radiolabeled, single-stranded cDNA probe containing the coxIII-c-myc junction demonstrated a nuclease-resistant band that matched the full length of the junctional cDNA probe. A smaller band that equaled the size of the c-myc portion alone was also detected. Only the smaller band coinciding with the c-myc sequences was protected from nuclease digestion by RNA from other cells. When a radiolabeled probe synthesized in the opposite orientation was used, nuclease-resistant bands equal in length to the coxIII portion of the probe were detected after hybridization with RNA from all cells. These results indicate that insertion of mtDNA fragments into nuclear genes occurs and that subsequent transcription of a 'chimaeric' or 'fusion' mRNA containing both mitochondrial and nuclear sequences can ensue.


Photoprotective and Antigenotoxic Effects of the Flavonoids Apigenin, Naringenin and Pinocembrin.
This work evaluated the photoprotective and antigenotoxic effects against ultraviolet B (UVB) radiation of flavonoid compounds apigenin, naringenin and pinocembrin. The photoprotective efficacy of these compounds was estimated using in vitro photoprotection indices, and the antigenotoxicity against UVB radiation was evaluated using the SOS chromotest and an enzymatic (proteinase K/T4 endonuclease V enzyme) comet assay in UV-treated Escherichia coli and human (HEK-293) cells, respectively. Naringenin and pinocembrin showed maximum UV-absorption peak in UVC and UVB zones, while apigenin showed UV-absorption capability from UVC to UVA range. These compounds acted as UV filters reducing UV-induced genotoxicity, both in bacteria and in human cells. The enzymatic comet assay resulted highly sensitive for detection of UVB-induced DNA damage in HEK-293 cells. In this work, the photoprotective potential of these flavonoids was widely discussed.


Unsuspected control of siderophore production by N-acetylglucosamine in streptomycetes.
Iron is one of the most abundant elements on earth but is found in poorly soluble forms hardly accessible to microorganisms. To subsist, they have developed iron-chelating molecules called siderophores that capture this element in the environment and the resulting complexes are internalized by specific uptake systems. While biosynthesis of siderophores in many bacteria is regulated by iron availability and oxidative stress, we describe here a new type of regulation of siderophore production. We show that in Streptomyces coelicolor, their production is also controlled by N-acetylglucosamine (GlcNAc) via the direct transcriptional repression of the iron utilization repressor dmdR1 by DasR, the GlcNAc utilization regulator. This regulatory nutrient-metal relationship is conserved among streptomycetes, which indicates that the link between GlcNAc utilization and iron uptake repression, however unsuspected, is the consequence of a successful evolutionary process. We describe here the molecular basis of a novel inhibitory mechanism of siderophore production that is independent of iron availability. We speculate that the regulatory connection between GlcNAc and siderophores might be associated with the competition for iron between streptomycetes and their fungal soil competitors, whose cell walls are built from the GlcNAc-containing polymer chitin. Alternatively, GlcNAc could emanate from streptomycetes' own peptidoglycan that goes through intense remodelling throughout their life cycle, thereby modulating the iron supply according to specific needs at different stages of their developmental programme.


Finding the extrema of a region.
The computational difficulty of finding the extreme points (those two points furthest from each other) in a region of an image is examined. Various approaches are described, including computation of convex hulls, sophisticated search strategies, and a statistical method. Suitability of the various methods to particular applications is discussed.


Stress as a state of anticipatory vigilance.
The pituitary-adrenocortical system is activated in stressful situations, and the adrenocortical hormones have been implicated by Selye in the diseases of stress. Evidence indicates that the adrenocortical hormones peak during the period of anticipation of stressful events, not during actual confrontation with them. The circadian cortisol rhythm shows peaks before waking, not during active periods of the day, and cortisol does not appear to be required during physical activity. The role of these hormones appears to be to suppress ongoing physiological activity in order to increase vigilance and to get the organism ready to take action before the impact of dangers.


High capacity binding of antibodies by poly(hydroxyethyl methacrylate) nanoparticles.
Poly(hydroxyethyl methacrylate) (PHEMA) nanoparticles with an average size of 150 nm in diameter and with a poly-dispersity index of 1.171 were produced by a surfactant free emulsion polymerization. Specific surface area of the PHEMA nanoparticles was found to be 1779 m(2)/g. Reactive imidazole containing 3-(2-imidazoline-1-yl)propyl(triethoxysilane) (IMEO) was used as a pseudo-specific ligand. IMEO was attached covalently onto the nanoparticles. PHEMA-IMEO nanoparticles were used for the affinity binding of immunoglobulin-G (IgG) from human plasma. To evaluate the degree of IMEO loading, the PHEMA nanoparticles were subjected to Si analysis by using flame atomizer atomic absorption spectrometer and it was estimated as 64.5 mg/g of polymer. The nanoparticles were characterized by transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FTIR). IgG binding onto the PHEMA nanoparticles was found to be 5.2 mg/g. Much higher binding values (up to 843 mg/g) were obtained for the PHEMA-IMEO nanoparticles. IgG could be repeatedly bound and eluted on PHEMA-IMEO nanoparticles without noticeable loss in the IgG binding capacity.


A Salmonella fim homologue in Citrobacter freundii mediates invasion in vitro and crossing of the blood-brain barrier in the rat pup model.
From the invasive Citrobacter freundii strain 3009, an invasion determinant was cloned, sequenced, and expressed. Sequence analysis of the determinant showed high homology with the fim determinant from Salmonella enterica serovar Typhimurium. The genes of the invasion determinant directed invasion of recombinant Escherichia coli K-12 strains into human epithelial cell lines of the bladder and gut as well as mannose-sensitive yeast agglutination and were termed fim(Cf) genes. Expression of the Fim(Cf) proteins was shown by (35)S labeling and/or Western blotting. In the infant rat model of experimental hematogenous meningitis, C. freundii strain 3009 and the in vitro invasive recombinant E. coli K-12 strain harboring the fim(Cf) determinant reached the cerebrospinal fluid, in contrast to the case for the control strain. The fim determinant was also necessary for efficient in vitro invasion by C. freundii, because a deletion mutant was strongly reduced in its invasion efficiency. The mutation could be complemented in trans by the corresponding genes. Invasion by C. freundii could be blocked only by d-mannose, GlcNAc, and chitin hydrolysate and not by other carbohydrates tested. In contrast, yeast agglutination was not affected by GlcNAc or chitin hydrolysate. This finding indicated mannose residues to be essential for both yeast agglutination and invasion, whereas GlcNAc (oligomer) residues of host cells are involved exclusively in invasion. These results showed the fim determinant of C. freundii to be responsible for d-mannose- and GlcNAc-dependent in vitro invasion without being assembled into pili and for crossing of the blood-brain barrier in the infant rat model.


Identification of a reduction product of aristolochic acid: implications for the metabolic activation of carcinogenic aristolochic acid.
Aristolochic acids are nephrotoxic and carcinogenic natural products that have been implicated both in endemic nephropathy in the Balkan region and in ailments caused by ingestion of herbal remedies. Aristolochic acids are metabolized to active intermediates that bind to DNA. In this study, reduction of aristolochic acid I with zinc in acetic acid afforded a new product that was characterized as 9-methoxy-7-methyl-2H-1,3-oxazolo[5',4'-10,9]phenanthro[3,4-d]-1,3-dioxolane-5-carboxylic acid, designated as aristoxazole, along with the expected aristolactam I. This new compound is a condensation product of aristolochic acid and acetic acid that may be related to the aristolochic acid-DNA adducts. The proposed mechanism of formation of aristoxazole involves nucleophilic attack of acetic acid on the nitrenium ion of aristolochic acid I. On the basis of these studies, a route to the metabolic activation of aristolochic acids and formation of adducts with DNA in in vitro systems is proposed and discussed.


Abdominal angina in patients receiving long-term hemodialysis.
The syndrome of abdominal angina is described in three patients receiving long-term hemodialysis. This clinical entity has not yet been described in patients with chronic renal failure. The classic triad of abdominal pain, weight loss, and diarrhea produces fear of eating. Most laboratory and radiologic investigations are inconclusive. Remarkable relief of symptoms occurs with maintenance of the hematocrit level above 30%, reduction of pump speed during hemodialysis to approximately 200 ml/min, use of a smaller surface area dialyzer, and administration of sublingual nitroglycerin. This clinical syndrome is often misdiagnosed, resulting in several unnecessary investigations, as well as unnecessary surgical procedures. Cholecystectomy is the operation most commonly done. All three of our patients died within ten months of diagnosis.


Crystal Structure and Functional Characterization of an Esterase (EaEST) from Exiguobacterium antarcticum.
A novel microbial esterase, EaEST, from a psychrophilic bacterium Exiguobacterium antarcticum B7, was identified and characterized. To our knowledge, this is the first report describing structural analysis and biochemical characterization of an esterase isolated from the genus Exiguobacterium. Crystal structure of EaEST, determined at a resolution of 1.9 Å, showed that the enzyme has a canonical α/β hydrolase fold with an α-helical cap domain and a catalytic triad consisting of Ser96, Asp220, and His248. Interestingly, the active site of the structure of EaEST is occupied by a peracetate molecule, which is the product of perhydrolysis of acetate. This result suggests that EaEST may have perhydrolase activity. The activity assay showed that EaEST has significant perhydrolase and esterase activity with respect to short-chain p-nitrophenyl esters (≤C8), naphthyl derivatives, phenyl acetate, and glyceryl tributyrate. However, the S96A single mutant had low esterase and perhydrolase activity. Moreover, the L27A mutant showed low levels of protein expression and solubility as well as preference for different substrates. On conducting an enantioselectivity analysis using R- and S-methyl-3-hydroxy-2-methylpropionate, a preference for R-enantiomers was observed. Surprisingly, immobilized EaEST was found to not only retain 200% of its initial activity after incubation for 1 h at 80°C, but also retained more than 60% of its initial activity after 20 cycles of reutilization. This research will serve as basis for future engineering of this esterase for biotechnological and industrial applications.


Effects of dynamic-demand-control appliances on the power grid frequency.
Power grid frequency control is a demanding task requiring expensive idle power plants to adapt the supply to the fluctuating demand. An alternative approach is controlling the demand side in such a way that certain appliances modify their operation to adapt to the power availability. This is especially important to achieve a high penetration of renewable energy sources. A number of methods to manage the demand side have been proposed. In this work we focus on dynamic demand control (DDC), where smart appliances can delay their switchings depending on the frequency of the system. We introduce a simple model to study the effects of DDC on the frequency of the power grid. The model includes the power plant equations, a stochastic model for the demand that reproduces, adjusting a single parameter, the statistical properties of frequency fluctuations measured experimentally, and a generic DDC protocol. We find that DDC can reduce small and medium-size fluctuations but it can also increase the probability of observing large frequency peaks due to the necessity of recovering pending task. We also conclude that a deployment of DDC around 30-40% already allows a significant reduction of the fluctuations while keeping the number of pending tasks low.


Measuring outcomes in healthcare economics using Artificial Intelligence: With application to resource management.
The quality of service in healthcare is constantly challenged by outlier events such as pandemics (i.e., Covid-19) and natural disasters (such as hurricanes and earthquakes). In most cases, such events lead to critical uncertainties in decision-making, as well as in multiple medical and economic aspects at a hospital. External (geographic) or internal factors (medical and managerial) lead to shifts in planning and budgeting, but most importantly, reduce confidence in conventional processes. In some cases, support from other hospitals proves necessary, which exacerbates the planning aspect. This paper presents three data-driven methods that provide data-driven indicators to help healthcare managers organize their economics and identify the most optimum plan for resources allocation and sharing. Conventional decision-making methods fall short in recommending validated policies for managers. Using reinforcement learning, genetic algorithms, traveling salesman, and clustering, we experimented with different healthcare variables and presented tools and outcomes that could be applied at health institutes. Experiments are performed; the results are recorded, evaluated, and presented.


Timing of initiation of antiretroviral drugs during tuberculosis therapy.
The rates of death are high among patients with coinfection with tuberculosis and the human immunodeficiency virus (HIV). The optimal timing for the initiation of antiretroviral therapy in relation to tuberculosis therapy remains controversial. In an open-label, randomized, controlled trial in Durban, South Africa, we assigned 642 patients with both tuberculosis and HIV infection to start antiretroviral therapy either during tuberculosis therapy (in two integrated-therapy groups) or after the completion of such treatment (in one sequential-therapy group). The diagnosis of tuberculosis was based on a positive sputum smear for acid-fast bacilli. Only patients with HIV infection and a CD4+ cell count of less than 500 per cubic millimeter were included. All patients received standard tuberculosis therapy, prophylaxis with trimethoprim-sulfamethoxazole, and a once-daily antiretroviral regimen of didanosine, lamivudine, and efavirenz. The primary end point was death from any cause. This analysis compares data from the sequential-therapy group and the combined integrated-therapy groups up to September 1, 2008, when the data and safety monitoring committee recommended that all patients receive integrated antiretroviral therapy. There was a reduction in the rate of death among the 429 patients in the combined integrated-therapy groups (5.4 deaths per 100 person-years, or 25 deaths), as compared with the 213 patients in the sequential-therapy group (12.1 per 100 person-years, or 27 deaths); a relative reduction of 56% (hazard ratio in the combined integrated-therapy groups, 0.44; 95% confidence interval, 0.25 to 0.79; P=0.003). Mortality was lower in the combined integrated-therapy groups in all CD4+ count strata. Rates of adverse events during follow-up were similar in the two study groups. The initiation of antiretroviral therapy during tuberculosis therapy significantly improved survival and provides further impetus for the integration of tuberculosis and HIV services. (ClinicalTrials.gov number, NCT00398996.)


Bat guano-dwelling microbes and antimicrobial properties of the pygidial gland secretion of a troglophilic ground beetle against them.
Bat guano is an important source of microbial diversity in caves and can be a source of potential pathogens. Laemostenus (Pristonychus) punctatus is a guanophilic ground beetle species, which pygidial gland secretion exhibits action against pathogenic and other microbes. The distribution and diversity of microbes in bat guano from a karstic cave were determined in this study. Additionally, antimicrobial activity of the pygidial gland secretion of L. (P.) punctatus against guano-dwelling microbes was tested; minimal inhibitory concentration (MIC) and chemical composition of the secretion were analyzed. In total, 63 different bacterial species and 16 fungal morphotypes were isolated from guano samples by the cultivation method and confirmed using and phenotypic characterization and molecular identification. There was a difference in the composition of certain microorganisms between the sampling points (cave locations) and between the guano layers. The largest number of bacterial isolates belongs to the genera Lysinibacillus and Paenibacillus, while Pseudomonas species were highly abundant at the innermost sampling point. For the guanophilic fungi, the majority are ascomycetes, with Penicillium and Aspergillus as the most dominant genera. Meyerozyma guilliermondii was the only yeast species found in the guano samples. The most sensitive isolates were Enterococcus eurekensis (MIC 0.007 mg/mL) and Escherichia fergusonii (MIC 0.028 mg/mL). The most sensitive fungal isolates were M. guilliermondii, Penicillium expansum, and Trichoderma harzianum (MIC 0.15 mg/mL). This study opens a new possibility for better understanding of ecological relations between microorganisms and troglophilic ground beetles and for detailed investigations of morpho-anatomical aspects of pygidial glands.


In vitro dosimetry of agglomerates.
Agglomeration of nanoparticles in biological fluids is a pervasive phenomenon that leads to difficulty in the interpretation of results from in vitro exposure, primarily due to differing particokinetics of agglomerates to nanoparticles. Therefore, well-defined small agglomerates were designed that possessed different particokinetic profiles, and their cellular uptake was compared to a computational model of dosimetry. The approach used here paves the way for a better understanding of the impact of agglomeration on the nanoparticle-cell interaction.


Enacting power and constructing gender in cervical cancer screening encounters between transmasculine patients and health care providers.
Transmasculine people are at risk of cervical cancer but have lower rates of cervical cancer screening than cisgender women. Disaffirmation of the patient's gender and unequal power dynamics between patient and provider during screening contribute to patient unwillingness to be screened. The mechanisms by which the balance of power may be shifted between patient and provider, and by which gender is constructed during the Pap test, are not well understood. A qualitative study using a modified grounded theory approach was undertaken to analyse patient interview and provider interview and focus group data pertaining to power and gender in the context of cervical cancer screening among transmasculine individuals. The study was conducted at an LGBTQ-focussed health centre in Boston, USA. Processes by which power is enacted included constraining or affirming patient choice, mitigating or exacerbating vulnerability, and self-advocacy. Gendering processes included naming patients and their bodies, invoking gender norms, de-gendering/re-gendering Pap tests, and othering or normalising trans bodies. The interplay between these processes promotes or constrains patient agency over body and health, impacting patient care, patient-provider interaction, and service utilisation. Understanding patient and provider roles in power and gender dynamics are critical for the provision of patient-centred care.


LexicHash: sequence similarity estimation via lexicographic comparison of hashes.
Pairwise sequence alignment is a heavy computational burden, particularly in the context of third-generation sequencing technologies. This issue is commonly addressed by approximately estimating sequence similarities using a hash-based method such as MinHash. In MinHash, all k-mers in a read are hashed and the minimum hash value, the min-hash, is stored. Pairwise similarities can then be estimated by counting the number of min-hash matches between a pair of reads, across many distinct hash functions. The choice of the parameter k controls an important tradeoff in the task of identifying alignments: larger k-values give greater confidence in the identification of alignments (high precision) but can lead to many missing alignments (low recall), particularly in the presence of significant noise. In this work, we introduce LexicHash, a new similarity estimation method that is effectively independent of the choice of k and attains the high precision of large-k and the high sensitivity of small-k MinHash. LexicHash is a variant of MinHash with a carefully designed hash function. When estimating the similarity between two reads, instead of simply checking whether min-hashes match (as in standard MinHash), one checks how "lexicographically similar" the LexicHash min-hashes are. In our experiments on 40 PacBio datasets, the area under the precision-recall curves obtained by LexicHash had an average improvement of 20.9% over MinHash. Additionally, the LexicHash framework lends itself naturally to an efficient search of the largest alignments, yielding an O(n) time algorithm, and circumventing the seemingly fundamental O(n2) scaling associated with pairwise similarity search. LexicHash is available on GitHub at https://github.com/gcgreenberg/LexicHash.


Is Cochlear Length Related to Congenital Sensorineural Hearing Loss: Preliminary Data.
This study used the data from patients with congenital sensorineural hearing loss (CSNHL) and those with normal hearing to measure and compare the length of the cochlea with high-resolution computed tomography (HRCT). HRCT images of patients who were diagnosed with CSNHL and were candidates for cochlear implantation were evaluated retrospectively. Sixty-three ears of 33 patients were included in the study. The control group comprised 66 ears of 33 individuals. The measurements were conducted by an experienced radiologist, using three-dimensional curved multiplanar reconstruction. All the measurements were performed thrice, and the average was calculated. The data were distributed normally. The lengths of the cochlear components for the CSNHL and control groups were as follows: basal turn 21.66±1.01 (21.30-22.02) and 22.57±0.68 (22.32-22.81) mm, middle turn 11.58±0.69 (11.34-11.83) and 12.39±0.46 (12.23-12.56) mm, and apical turn 6.45±0.92 (6.12-6.77) and 7.12±0.65 (6.89-7.35) mm, respectively. The mean cochlear lateral wall (LW) length was significantly shorter in the CSNHL patients [39.71±1.32 (39.25-40.18) mm] than in the controls [42.09±1.17 (41.67-42.51) mm], (p<0.001). The intra-rater reliability was 0.878 (confidence interval 95%: 0.841-0.908 p<0.001). The cut-off value was 40.81 mm (sensitivity: 0.91, specificity: 0.94, and accuracy: 0.90). There were microanatomic dissimilarities between the length of the cochlea in subjects from the CSNHL group and those from the control group.


Emergence of Influenza Viruses and Crossing the Species Barrier.
Influenza A viruses are zoonotic pathogens that infect a variety of host species including wild aquatic birds, domestic poultry, and a limited number of mammals including humans. The error-prone nature of the virus's replication machinery and its ability to transmit among multiple hosts lead to generation of novel virus variants with altered pathogenicity and virulence. Spatial, molecular, and physiological barriers inhibit cross-species infections, particularly in the case of human infection with avian viruses. Pigs are proposed as a mixing vessel that facilitates movement of avian viruses from the wild bird reservoir into humans. However, the past decade has witnessed the emergence of highly pathogenic and virulent avian H5 and H7 viruses that have breached these barriers, bypassed the pig intermediate host, and infected humans with a high mortality rate, but have not established human-to-human transmissible lineages. Because influenza viruses pose a significant risk to both human and animal health, it is becoming increasingly important to attempt to predict their identities and pathogenic potential before their widespread emergence. Surveillance of the wild bird reservoir, molecular characterization and documentation of currently circulating viruses in humans and animals, and a comprehensive risk assessment analysis of individual isolates should remain a high priority. Such efforts are critical to the pursuit of prevention and control strategies, including vaccine development and assessment of antiviral susceptibility, that will have a direct impact on the well-being of humans and animals worldwide.


Upregulation of E2F transcription factor 3 is associated with poor prognosis in hepatocellular carcinoma.
E2F transcription factor 3 (E2F3), a member of the E2F transcription factor family and a member of the genes involved in the regulation of cell cycle, is an oncogene with strong proliferative potential. E2F3 is involved in many processes and plays important roles in the development of several types of cancer, while its relationship with prognosis in hepatocellular carcinoma (HCC) has yet to be reported. In the present study, based on 4 independent microarray data sets which covered 385 cases of HCC and 327 cases of normal livers retrieved from the Oncomine database, we demonstrated that E2F3 was upregulated at least 1.5-fold and on average 2.3-fold in HCC when compared with normal controls. Comprehensive bioinformatics analysis consisting of protein-protein interaction, gene co-occurrence, microRNA-mRNA interaction and biological process annotation indicated that E2F3 interacted with a large number of genes, proteins and microRNAs which were all associated with poor prognosis in patients with HCC and other types of cancer, suggesting that E2F3 may also serve as a biomarker for poor prognosis. Taken together, for the first time, we show that the overexpression of E2F3 may be associated with unfavorable prognosis in HCC.


Changes in low molecular weight phenolic compounds in Spanish, French, and American oak woods during natural seasoning and toasting.
The evolution of low molecular weight polyphenols in Spanish oak heartwood of Quercus robur,Quercus petraea, Quercus pyrenaica, and Quercus faginea was studied by HPLC, in relation to the processing of wood in barrel cooperage. The polyphenolic composition of Spanish woods subjected to natural seasoning for 3 years and to the toasting process was studied in relation to those of French oak of Q. robur (Limousin) and Q. petraea (Allier) and American oak of Q. alba (Missouri), which are habitually used in cooperage. The concentrations of benzoic and cinnamic acids and aldehydes of Spanish woods increased during seasoning depending on the duration of this process and in the same way as those of French and American woods. The process having the main influence on the phenolic composition of wood was the toasting. It led to high increases in the concentration of phenolic aldehydes and acids, especially cinnamic aldehydes (sinapic and coniferylic aldehydes), followed by benzoic aldehydes (syringaldehyde and vanillin) and benzoic acids (syringic and vanillic acids). This polyphenolic composition in Spanish oak species evolved during toasting as in French and American oak, but quantitative differences were found, which were especially important in American species with respect to the others.


Characterization of two epiphytic bacteria from soybean leaves with antagonistic activities against Pseudomonas syringae pv. glycinea.
The strains 48b/90 and 22d/93 are naturally occurring ephiphytes which were isolated from soybean leaves. On the basis of pheno- and genotypic characteristics 48b/90 was identified as Erwinia herbicola and 22d/93 as Pseudomonas syringae. These two isolates produced biological active substances against different indicator organisms. The E. herbicola strain showed clear antagonistic properties against Escherichia coli and Pseudomonas syringae pv. glycinea, but not against Geotrichum candidum. 22d/93 was active against P. glycinea and G. candidum, but not against E. coli. Strain 48b/90 produced at least two different inhibitors: an antibiotic substance and an inhibitor of the alginate synthesis. Strain 22d/93 produced at least three different compounds inhibitory to P. glycinea and one to G. candidum. Their activities against the bacterial blight pathogen, P. glycinea, can be observed in planta, too. Under the influences of the antagonists the pathogen multiplied at lower rates and to lower stationary phase population levels. The development of bacterial blight symptoms was suppressed.


A pilot study of medical student attitudes to, and use of, commercial movies that address public health issues.
An innovative approach to learning public health by using feature-length commercial movies was piloted in the fourth year of a medical degree. We aimed to explore how students responded to this approach and the relative effectiveness of two promotional strategies. Firstly we placed DVDs of 15 movies (with public health-related content) in the medical school library. Then alternating groups of students (total n = 82 students) were exposed to either a brief promotional intervention or a more intensive intervention involving a class presentation. The response rates were 99% at baseline and 85% at follow-up. The level and strength of support for using movies in public health training increased after exposure to the public health module with significantly more students "strongly agreeing". Student behaviour, in terms of movies viewed or accessed from the library, also suggested student interest. While there were no statistically significant differences in median viewing or library access rates between the two intervention groups, the distribution of viewing patterns was shifted favourably. Those exposed to the more intensive intervention (class presentation) were significantly more likely to have reported watching at least one movie (97% vs. 81%; p = 0.033) or to having accessed at least one movie from the library (100% vs. 70%, p = 0.0001). This pilot study found that the students had very positive attitudes towards viewing public health-related commercial movies. Movie access rates from the library were also favourable.


Fake news on Social Media: the Impact on Society.
Fake news (FN) on social media (SM) rose to prominence in 2016 during the United States of America presidential election, leading people to question science, true news (TN), and societal norms. FN is increasingly affecting societal values, changing opinions on critical issues and topics as well as redefining facts, truths, and beliefs. To understand the degree to which FN has changed society and the meaning of FN, this study proposes a novel conceptual framework derived from the literature on FN, SM, and societal acceptance theory. The conceptual framework is developed into a meta-framework that analyzes survey data from 356 respondents. This study explored fuzzy set-theoretic comparative analysis; the outcomes of this research suggest that societies are split on differentiating TN from FN. The results also show splits in societal values. Overall, this study provides a new perspective on how FN on SM is disintegrating societies and replacing TN with FN.


DNA double-strand break repair functions defend against parvovirus infection.
We measured parvovirus replication and sensitivity to X-ray damage in nine CHO cell lines representing a variety of DNA repair deficiencies. We found that parvovirus replication efficiency increases with radiosensitivity. Parvovirus replication is disrupted at an early stage of infection in DNA repair-proficient cells, before conversion of the single-stranded viral DNA genome into the double-stranded replicative form. Thus, status of the DNA repair machinery inversely correlates with parvovirus replication and is proportional to the host's ability to repair X-ray-induced damage.


Taylor spatial frame in severe foot deformities using double osteotomy: technical approach and primary results.
The treatment of severe foot deformities in children or adolescents is complex and demanding to the surgeon. This paper describes the technical strategy of using a Taylor spatial frame and reports on the functional outcome. The Taylor spatial frame was used by a single surgeon in patients with severe foot deformities. Seven patients with eight severe deformities were treated. Mean patient age at surgery was 15.1 (9-29) years. A double Taylor spatial frame reconstruction was mounted to the limb. All patients had a midtarsal osteotomy and an additional inverse dome-shaped calcaneus osteotomy. Assessed measures were pre- and postoperative deformity and associated complications and clinical results according to Ferreira et al. Mean follow-up was 576.5 (359-987) days. The final functional outcome according to Ferreira was good in seven cases, fair in none, and poor in one. Early complications included pin-tract infection, temporary hypoesthesia, and temporary shortening of the tendon of the M. flexor digitorum. At follow-up there was no deformity recurrence. In children or adolescents, the innovative treatment using the Taylor spatial frame and a double osteotomy allows joint-preserving correction of severe foot deformities. However, the complication rate is relatively high due to the severely deformed feet. Furthermore, the Taylor spatial frame is expensive. Thus, this treatment is only recommended for severe foot deformities and should be handled by experienced orthopedic surgeons.


GRWD1, a new player among oncogenesis-related ribosomal/nucleolar proteins.
Increasing attention has been paid to certain ribosomal or ribosome biosynthesis-related proteins involved in oncogenesis. Members of one group are classified as "tumor suppressive factors" represented by RPL5 and RPL11; loss of their functions leads to cancer predisposition. RPL5 and RPL11 prevent tumorigenesis by binding to and inhibiting the MDM2 ubiquitin ligase and thereby up-regulating p53. Many other candidate tumor suppressive ribosomal/nucleolar proteins have been suggested. However, it remains to be experimentally clarified whether many of these factors can actually prevent tumorigenesis and if so, how they do so. Conversely, some ribosomal/nucleolar proteins promote tumorigenesis. For example, PICT1 binds to and anchors RPL11 in nucleoli, down-regulating p53 and promoting tumorigenesis. GRWD1 was recently identified as another such factor. When overexpressed, GRWD1 suppresses p53 and transforms normal human cells, probably by binding to RPL11 and sequestrating it from MDM2. However, other pathways may also be involved.


Accelerating AXL targeting for TNBC therapy.
The tyrosine kinase receptor AXL of the TAM (TYRO3, AXL and MERTK) family is considered as a promising therapeutic target for different hematological cancers and solid tumors. AXL is involved in multiple pro-tumorigenic processes including cell migration, invasion, epithelial-mesenchymal transition (EMT), and stemness, and recent studies demonstrated its impact on cancer metastasis and drug resistance. Extensive studies on AXL have highlighted its unique characteristics and physiological functions and suggest that targeting of AXL could be beneficial in combination with chemotherapy, radiotherapy, immunotherapy, and targeted therapy. In this mini review, we discuss possible outcomes of AXL targeting either alone or together with other therapeutic agents and emphasize its impact on triple negative breast cancer (TNBC).


A col10a1:nlGFP transgenic line displays putative osteoblast precursors at the medaka notochordal sheath prior to mineralization.
In teleosts, such as medaka, ossification of the vertebral column starts with the mineralization of the notochordal sheath in a segmental pattern. This establishes the chordal centrum, which serves as the basis for further ossifications by sclerotome derived osteoblasts generating the vertebral body. So far, it is unclear which cells produce the notochordal sheath and how a segmental pattern of mineralization is established in teleosts. Here, we use a transgenic medaka line that expresses nlGFP under the control of the col10a1 promoter for in vivo analysis of vertebral body formation. We show that col10a1:nlGFP expression recapitulates endogenous col10a1 expression. In the axial skeleton, col10a1:nlGFP cells appear prior to the mineralization of the notochordal sheath in a segmental pattern. These cells remain on the outer surface of the chordal centra during mineralization as well as subsequent perichordal ossification of the vertebral bodies. Using twist1a1:dsRed and osx:mCherry transgenic lines we show that a subset of col10a1:nlGFP cells is derived from sclerotomal precursors and differentiates into future osteoblasts. For the first time, this shows a segmental occurrence of putative osteoblast precursors in the vertebral centra prior to ossification of the notochordal sheath. This opens the possibility that sclerotome derived cells in teleosts are implicated in the establishment of the mineralized vertebral column in a similar manner as previously described for tetrapods.


Metaecosystem Dynamics of Marine Phytoplankton Alters Resource Use Efficiency along Stoichiometric Gradients.
Metaecosystem theory addresses the link between local (within habitats) and regional (between habitats) dynamics by simultaneously analyzing spatial community ecology and abiotic matter flow. Here we experimentally address how spatial resource gradients and connectivity affect resource use efficiency (RUE) and stoichiometry in marine phytoplankton as well as the community composition at local and regional scales. We created gradostat metaecosystems consisting of five linearly interconnected patches, which were arranged either in countercurrent gradients of nitrogen (N) and phosphorus (P) supply or with a uniform spatial distribution of nutrients and which had either low or high connectivity. Gradient metaecosystems were characterized by higher remaining N and P concentrations (and N∶P ratios) than uniform ones, a difference reduced by higher connectivity. The position of the patch in the gradient strongly constrained elemental stoichiometry, local biovolume production, and RUE. As expected, algal carbon (C)∶N, biovolume, and N-specific RUE decreased toward the N-rich end of the gradient metaecosystem, whereas the opposite was observed for most of the gradient for C∶P, N∶P, and P-specific RUE. However, at highest N∶P supply, unexpectedly low C∶P, N∶P, and P-specific RUE values were found, indicating that the low availability of P inhibited efficient use of N and biovolume production. Consequently, gradient metaecosystems had lower overall biovolume at the regional scale. Whereas treatment effects on local richness were weak, gradients were characterized by higher dissimilarity in species composition. Thus, the stoichiometry of resource supply and spatial connectivity between patches appeared as decisive elements constraining phytoplankton composition and functioning in metaecosystems.


Integrating in silico prediction methods, molecular docking, and molecular dynamics simulation to predict the impact of ALK missense mutations in structural perspective.
Over the past decade, advancements in next generation sequencing technology have placed personalized genomic medicine upon horizon. Understanding the likelihood of disease causing mutations in complex diseases as pathogenic or neutral remains as a major task and even impossible in the structural context because of its time consuming and expensive experiments. Among the various diseases causing mutations, single nucleotide polymorphisms (SNPs) play a vital role in defining individual's susceptibility to disease and drug response. Understanding the genotype-phenotype relationship through SNPs is the first and most important step in drug research and development. Detailed understanding of the effect of SNPs on patient drug response is a key factor in the establishment of personalized medicine. In this paper, we represent a computational pipeline in anaplastic lymphoma kinase (ALK) for SNP-centred study by the application of in silico prediction methods, molecular docking, and molecular dynamics simulation approaches. Combination of computational methods provides a way in understanding the impact of deleterious mutations in altering the protein drug targets and eventually leading to variable patient's drug response. We hope this rapid and cost effective pipeline will also serve as a bridge to connect the clinicians and in silico resources in tailoring treatments to the patients' specific genotype.


Implementation of a cloud-based electronic patient-reported outcome (ePRO) platform in patients with advanced cancer.
Patient reported outcomes (PROs) have been associated with improved symptom management and quality of life in patients with cancer. However, the implementation of PROs in an academic clinical practice has not been thoroughly described. Here we report on the execution, feasibility and healthcare utilization outcomes of an electronic PRO (ePRO) application for cancer patients at an academic medical center. We conducted a randomized trial comparing an experimental ePRO arm to standard of care in patients with advanced cancer in the thoracic, gastrointestinal, and genitourinary oncology groups at Stanford Cancer Center from March 2018 to November 2019. We describe the pre-implementation, implementation, and post-implementation phases of the ePRO arm, technological barriers, electronic health record (EHR) integration, clinician burden, and patient data privacy and security. Feasibility was pre-specified to be at least 70% completion of all questionnaires. Acceptability was based on patient and clinician feedback. Ambulatory healthcare utilization was assessed by reviewing numbers of phone messages, electronic portal messages, and referrals for supportive care. Of 617 ePRO questionnaires sent to 72 patients, 445 (72%) were completed. Most clinicians (87.5%) and patients (93%) felt neutral or positive about the ePRO tool's ease of use. Exposure to ePRO did not cause a measurable change in ambulatory healthcare utilization, with a median of less than two phone messages and supportive care referrals, and 5-6 portal messages. Web-based ePRO tools for patients with advanced cancer are feasible and acceptable without increasing clinical burden. Key lessons include the importance of pilot testing, engagement of stakeholders at all levels, and the need for customization by disease group. Future directions for this work include completion of EHR integration, expansion to other centers, and development of integrated workflows for routine clinical practice.


Transactional Patterns of Maternal Depressive Symptoms and Mother-Child Mutual Negativity in an Adoption Sample.
Transactional models of analysis can examine both moment-to-moment interactions within a dyad and dyadic patterns of influence across time. This study used data from a prospective adoption study to test a transactional model of parental depressive symptoms and mutual negativity between mother and child over time, utilizing contingency analysis of second-by-second behavioral data. To consider both genetic and environmental influences on mutual negativity, depressive symptoms were examined in both adoptive and birth mothers. Adoptive mother depressive symptoms at 9 months increased the likelihood that, at 18 months, children reacted negatively to their mothers' negative behavior, which in turn predicted higher levels of adoptive mother depressive symptoms at 27 months, suggesting that over time, mothers' depressive symptoms influence and are influenced by moment-to-moment mutual negativity with their toddlers. Birth mother depressive symptoms moderated the association between mutual negativity at 18 months and adoptive mother depressive symptoms at 27 months, suggesting a child-driven contribution to maternal depressive symptoms that can be measured by a genetic sensitivity.


The Zoonoses Order, 1975.
The coming into effect on July 14 of the Zoonoses Order 1975, marks a new and important step in veterinary public health. The Order makes available powers to investigate and control Salmonella and Brucella infections in animals and Salmonella infections in birds which present a threat to human health. Any Salmonella and Brucella infection becomes reportable in certaain species when organisms have been isolated by laboratory test. Veterinary surgeons and livestock owners may be involved in this obligatory procedure. The statutory powers to investigate incidents and apply restrictions on the movement of animals, birds and their products are not limited to reportable species: they cover a much wider range of animals and birds. They will be held in reserve and used only when necessary because of a hazard to public health; the use of restrictions will be limited to cases involving a very serious threat to man. The information collected from reports and investigations will add considerably to the epidemiological knowledge of these infections and so allow improved methods of control to be developed. Some of these measures already in operation are described, together with others possible for the future.


GCM1 regulation of the expression of syncytin 2 and its cognate receptor MFSD2A in human placenta.
Syncytin 2 is a newly identified placental membrane protein with fusogenic and immunosuppressive activities. Major facilitator superfamily domain containing 2A (MFSD2A) is the cognate receptor for syncytin 2-mediated cell-cell fusion. Both syncytin 2 and MFSD2A are highly expressed in placenta. In this study to understand the regulation of syncytin 2 and MFSD2A expression in placenta, we found that syncytin 2 gene is epigenetically silenced in nonplacental cells by cytosine-phosphate-guanine (CpG) dinucleotide methylation and that expression of syncytin 2 and MFSD2A genes are regulated by the placental transcription factor GCM1 in placental cells. Functional GCM1-binding sites were identified in syncytin 2 and MFSD2A promoters based on electrophoretic mobility shift assay and chromatin immunoprecipitation assay. Because GCM1 activity is decreased in hypoxic placental cells, we further confirmed that expression of MFSD2A is downregulated in hypoxic BeWo choriocarcinoma cells. Interestingly, ectopic expression of GCM1 activated syncytin 2 and MFSD2A expression in MCF-7 breast cancer cells and facilitated MCF-7 cell fusion. The expression of syncytin 2 in MCF-7 cells was partly attributed to CpG demethylation in the syncytin 2 promoter in the presence of GCM1. Our results suggest that GCM1 is a critical factor in controlling placental cell fusion through transcriptional regulation of syncytin 2 and MFSD2A gene expression in placenta. In addition, GCM1 may also play an important role in the epigenetic regulation of syncytin 2 gene expression.


Nanocarrier of α-Tocopheryl Succinate Based on a Copolymer Derivative of (4,7-dichloroquinolin-2-yl)methanol and Its Cytotoxicity against a Breast Cancer Cell Line.
In order to improve the water solubility and, therefore, bioavailability and therapeutic activity of anticancer hydrophobic drug α-tocopherol succinate (α-TOS), in this work, copolymers were synthesized via free radicals from QMES (1-[4,7-dichloroquinolin-2-ylmethyl]-4-methacryloyloxyethyl succinate) and VP (N-vinyl-2-pirrolidone) using different molar ratios, and were used to nanoencapsulate and deliver α-TOS into cancer cells MCF-7. QMES monomer was chosen because the QMES pendant group in the polymer tends to hydrolyze to form free 4,7-dichloro-2-quinolinemethanol (QOH), which also, like α-TOS, exhibit anti-proliferative effects on cancerous cells. From the QMES-VP 30:70 (QMES-30) and 40:60 (QMES-40) copolymers obtained, it was possible to prepare aqueous suspensions of empty nanoparticles (NPs) loaded with α-TOS by nanoprecipitation. The diameter and encapsulation efficiency (%EE) of the QMES-30 NPs loaded with α-TOS were 128.6 nm and 52%; while for the QMES-40 NPs loaded with α-TOS, they were 148.8 nm and 65%. The results of the AlamarBlue assay at 72 h of treatment show that empty QMES-30 NPs (without α-TOS) produced a marked cytotoxic effect on MCF-7 breast cancer cells, corresponding to an IC50 value of 0.043 mg mL-1, and importantly, they did not exhibit cytotoxicity against healthy HUVEC cells. Furthermore, NP-QMES-40 loaded with α-TOS were cytotoxic with an IC50 value of 0.076 mg mL-1, demonstrating a progressive release of α-TOS; however, the latter nanoparticles were also cytotoxic to healthy cells in the range of the assayed concentrations. These results contribute to the search for a new polymeric nanocarrier of QOH, α-TOS or other hydrophobic drugs for the treatment of cancer or others diseases treatable with these drugs.


Lung cancer-reacting human recombinant antibody AE6F4: potential usefulness in the sputum cytodiagnosis.
Human monoclonal antibody (hMAb) AE6F4 has been shown to be potentially useful for immunocytological detection of lung cancer cells in sputum. By recombinant DNA technology, IgM type hMAb AE6F4 was switched to lgG. The IgG mimic recombinant AE6F4 antibody expression plasmid was assembled using the antibody heavy chain gene, which ligated the gene encoding VH and CH1(mu) domains of hMAb AE6F4 heavy chain to the gene encoding CH2(gamma 1) and CH3(gamma 1) domains of human IgG heavy chain, and the antibody light chain gene of hMAb AE6F4. The recombinant antibody expressed by baby hamster kidney (BHK)-21 cells showed molecular size equivalence to IgG, and consisted of human mu-gamma hybrid heavy and kappa light chains. The immunological specificity of the recombinant antibody was the same as that of hMAb AE6F4 by immunoblotting analysis to the 14-3-3 protein, the putative antigen of hMAb AE6F4, and by immunohistochemical and immunocytological analyses using tissue sections and sputa of lung cancer patients. The transfected BHK-21 cells produced the recombinant antibody persistently and the productivity was greater than 20 times that by human-human hybridoma producing hMAb AE6F4.


Frequent allopolyploidy with distant progenitors in the moss genera Physcomitrium and Entosthodon (Funariaceae) identified via subgenome phasing of targeted nuclear genes.
Allopolyploids represent a new frontier in species discovery among embryophytes. Within mosses, allopolyploid discovery is challenged by low morphological complexity. The rapid expansion of sequencing approaches in addition to computational developments to identifying genome merger and whole-genome duplication using variation among nuclear loci representing homeologs has allowed for increased allopolyploid discovery among mosses. Here, we test a novel approach to phasing homeologs within loci and phasing loci across subgenomes, or subgenome assignment, called Homologizer, in the family Funariaceae. We confirm the intergeneric hybrid nature of Entosthodon hungaricus, and the allopolyploid origin of Physcomitrium eurystomum and one population of Physcomitrium collenchymatum. We also reveal that hybridization gave rise to Physcomitrium immersum, as well as to yet unrecognized lineages sharing the phenotype of Physcomitrium pyriforme and Physcomitrium sphaericum. Our findings demonstrate the utility of our approach when working with polyploid genomes, and its value in identifying progenitor species using target capture data.


Prospective study of alcohol use and hearing loss in men.
Hearing loss is a common and disabling sensory disorder, yet prospective data on potentially modifiable risk factors are limited. Previous studies suggest that alcohol consumption may influence the development of hearing loss, yet results have been inconsistent. The purpose of this study was to prospectively examine the relation between alcohol use and hearing loss in men. We examined prospectively the independent association between alcohol intake and self-reported professionally diagnosed hearing loss in 26,809 men aged 40 to 74 yrs at baseline in 1986. Study participants completed detailed questionnaires at baseline and every 2 yrs thereafter. Incident cases of hearing loss were defined as those professionally diagnosed after 1986. Cox proportional hazards multivariate regression was used to adjust for potential confounding factors. During 386,081 person-years of follow-up, 3447 incident cases of hearing loss were reported. Overall, there was no association between level of alcohol intake and risk of hearing loss. Compared with those who did not consume alcohol, the multivariate-adjusted hazard ratios (95% confidence interval) were 1.00 (0.89 to 1.12) for those who consumed 5.0 to 9.9 g/day, 1.08 (0.96 to 1.21) for 10.0 to 14.9 g/day, and 0.98 (0.85 to 1.13) for 30.0 to 49.9 g/day. The results did not differ by age group or folate intake. Among those with lower intake of vitamin B12, however, higher consumption of alcohol, specifically liquor, was associated with an increased risk of hearing loss. Our data suggest that low or moderate alcohol consumption does not influence the risk of hearing loss in older men. A possible relation between vitamin B12 intake, alcohol consumption, and hearing loss merits further investigation.


Does Increasing Patient Complexity Have an Effect on Medical Outcomes and Lengths-of-Stay after Total Knee Arthroplasty?
A greater number of medically complex patients with multiple comorbidities are now more readily considered for total knee arthroplasty (TKA). Therefore, the purpose of this study was to determine whether comorbidity burden, measured with the Elixhauser Comorbidity Index (ECI), correlated with 90-day medical complications and longer in-hospital lengths-of-stay (LOS) in TKA patients. The PearlDiver supercomputer was queried for all primary TKA patients in the Medicare Standard Analytic Files from 2005 to 2014 using International Classification of Disease, 9th edition codes. Patients were included based on ECI scores, ranging from 1 to 5. ECI 1 patients served as the control cohort, while ECI 2, 3, 4, and 5 patients were considered study cohorts. Each study cohort was matched based on age and gender to the control cohort, resulting in a total of 715,398 patients included for analysis (ECI 1, n = 144,072; ECI 2, n = 144,072; ECI 3, n = 144,072; ECI 4, n = 144,072; ECI 5, n = 139,110). Logistic regression analyses were performed to compare 90-day medical complications and Welch's t-tests were performed to compare LOS between the cohorts. Patients with higher ECI scores were more likely to develop medical complications and have longer LOS compared with matched patients in the control cohort. Compared with matched ECI 1 patients, patients with ECI scores of 2 (odds ratio [OR]: 1.19, 95% confidence interval [CI]: 1.14-1.24), 3 (OR: 1.27, 95% CI: 1.21-1.32), 4 (OR: 1.32, 95% CI: 1.27-1.38), and 5 (OR: 1.33, 95% CI: 1.27-1.39) were significantly more likely to develop 90-day medical complications. Additionally, the mean LOS of patients in the ECI 2 (2.59 ± 1.49 vs. 2.73 ± 1.52 days), ECI 3 (2.59 ± 1.49 vs. 2.88 ± 1.51 days; p < 0.001), ECI 4 (2.59 ± 1.49 vs. 3.01 ± 1.56 days; p < 0.001), and ECI 5 (2.61 ± 1.49 vs. 3.14 ± 1.61 days; p < 0.001) groups were significantly longer than the mean LOS in the control ECI 1 group. In an increasingly complex patient population, associations between comorbidities and outcomes after TKA procedures can guide providers on how to modify their pre- and postoperative care. These results demonstrate that higher ECI scores are associated with a greater likelihood of 90-day medical complications and longer in-hospital LOS.


Neurocircuits underlying cognition-emotion interaction in a social decision making context.
Decision making (DM) in the context of others often entails complex cognition-emotion interaction. While the literature suggests that the ventromedial prefrontal cortex (vmPFC), striatum, and amygdala are involved in valuation-based DM and hippocampus in context processing, how these neural mechanisms subserve the integration of cognitive and emotional values in a social context remains unclear. In this study we addressed this gap by systematically manipulating cognition-emotion interaction in a social DM context, when the participants played a card game with a hypothetical opponent in a behavioral study (n=73) and a functional magnetic-resonance-imaging study (n=16). We observed that payoff-based behavioral choices were influenced by emotional values carried by face pictures and identified neurocircuits involved in cognitive valuation, emotional valuation, and concurrent cognition-emotion value integration. Specifically, while the vmPFC, amygdala, and ventral striatum were all involved in both cognitive and emotional domains of valuation, these regions played dissociable roles in social DM. The payoff-dependent responses in vmPFC and amygdala, but not ventral striatum, were moderated by the social context. Furthermore, the vmPFC, but not amygdala, not only encoded the opponent's gains as if self's losses, but also represented a "final common currency" during valuation-based decisions. The extent to which emotional input influenced choices was associated with the functional connectivity between the value-signaling amygdala and value integrating vmPFC, and also with the functional connectivity between the context-setting hippocampus and value-signaling amygdala and ventral striatum. These results identify brain pathways through which emotion shapes subjective values in a social DM context.


A procedure for obtaining thin sections of undercalcified bone biopsies embedded in methyl methacrylate.
A complex of techniques is described, including a specially designed mould and a method for flat mounting of the sections, enabling to obtain well-stretched and undamaged thin sections of undecalcified bone biopsies. Apart from a gain in polymerisation time, a clear advantage of the special mould is a more favorable temperature course in and around the tissue during the polymerisation process.


Chiari 1000 Registry Project: assessment of surgical outcome on self-focused attention, pain, and delayed recall.
Prior research has typically found a negative relationship between chronic pain and memory, and we examined whether cognitive control processes (e.g. reflection and rumination) moderated this relationship in individuals with Chiari malformation Type I (CM). CM is a neurological condition in which the cerebellar tonsils descend into the medullary and upper cervical spine regions potentially resulting in severe headaches and neck pain. CM patients who had (n = 341) and had not (n = 297) undergone decompression surgery completed the McGill Pain Questionnaire-Short Form-Revised (SF-MPQ-2), the Rey Auditory Verbal Learning Test (RAVLT), and the Rumination-Reflection Questionnaire (RRQ). Immediate recall scores were compared to those of 102 healthy controls, and delayed recall performance was compared across other variables within the CM group. CM patients performed more poorly on immediate recall than did controls. Within CM patients, we observed main effects for reflection and age, and a pain x reflection x surgical status (surgery v. no surgery) interaction in which non-decompressed individuals with low levels of pain and high levels of reflection showed superior delayed recall relative to non-decompressed individuals with higher pain and all decompressed individuals. CM patients show an immediate recall deficit relative to controls, regardless of surgical status. High levels of reflection were associated with better delayed recall performance in non-decompressed CM patients with lower pain levels. High levels of chronic pain may overwhelm increased focused attention abilities, but higher levels of reflection partially overcome the distracting effects of pain and this may represent a type of resilience.


Heart rate-based lactate minimum test: a reproducible method.
To find the individual intensity for aerobic endurance training, the lactate minimum test (LMT) seems to be a promising method. LMTs described in the literature consist of speed or work rate-based protocols, but for training prescription in daily practice mostly heart rate is used. The aim of the present study was to investigate the reproducibility of a new heart rate-based LMT protocol. 20 subjects each underwent four LMTs on a cycle ergometer. The LMT consisted of a first part (Conconi test) to induce lactate accumulation and a second part (incremental protocol) with stages of 5 min starting beyond the aerobic threshold. During these stages work rate was adjusted to reach predetermined heart rates, which were calculated for every single test from heart rate (HR) at rest and maximum HR. Lactate was measured after each stage to define the lactate minimum (LM) and the corresponding HR. Institute of Sports Medicine. 20 healthy and endurance-trained individuals (13 men, seven women). Reproducibility of heart rate at LM. The reproducibility of heart rate at LM was high (coefficient of variation (CV) = 2.1%). The reproducibility of work rate at LM was good (CV = 6.7%). CV for lactate concentrations at LM was 17.4%. High interindividual differences at LM were found in heart rate (range 149-178 beats/min) and lactate levels (range 1.2-6.8 mmol/l). The LMT using a heart rate-based protocol is a reproducible method of assessing HR at an exercise intensity where an equilibrium exists between blood lactate accumulation and elimination.


Calcium phosphates: what is the evidence?
A number of different calcium phosphate compounds such as calcium phosphate cements and solid beta-tricalcium phosphate products have been introduced during the last decade. The chemical composition mimics the mineral phase of bone and as a result of this likeness, the materials seem to be remodeled as for normal bone through a cell-mediated process that involves osteoclastic activity. This is a major difference when compared with, for instance, calcium sulphate compounds that after implantation dissolve irrespective of the new bone formation rate. Calcium phosphates are highly biocompatible and in addition, they act as synthetic osteoconductive scaffolds after implantation in bone. When placed adjacent to bone, osteoid is formed directly on the surface of the calcium phosphate with no soft tissue interposed. Remodeling is slow and incomplete, but by adding more and larger pores, like in ultraporous beta-tricalcium phosphate, complete or nearly complete resorption can be achieved. The indications explored so far include filling of metaphyseal fracture voids or bone cysts, a volume expander in conjunction with inductive products, and as a carrier for various growth factors and antibiotics. Calcium phosphate compounds such as calcium phosphate cement and beta-tricalcium phosphate will most certainly be part of the future armamentarium when dealing with fracture treatment. It is reasonable to believe that we have so far only seen the beginning when it comes to clinical applications.


New insights into the mechanisms controlling the bronchial mucus balance.
In this work, we aim to analyze and compare the mechanisms controlling the volume of mucus in the bronchial region of the lungs of a healthy human adult, at rest and in usual atmospheric conditions. This analysis is based on a balance equation for the mucus in an airway, completed by a computational tool aiming at characterizing the evaporation, during respiration, of the water contained in the bronchial mucus. An idealized representation of the lungs, based on Weibel's morphometric model, is used. The results indicate that the mechanisms controlling the volume of mucus in an airway depend on the localization of the airway in the bronchial region of the lungs. In the proximal generations, the volume of mucus in an airway is mainly controlled by the evaporation of the water it contains and the replenishment, with water, of the mucus layer by epithelial cells or the submucosal glands. Nevertheless, cilia beating in this part of the bronchial region remains of fundamental importance to transport the mucus and hence to eliminate dust and pathogens trapped in it. On the other hand, in the distal generations of the bronchial region, the volume of mucus in an airway is mainly controlled by the mucociliary transport and by the absorption of liquid by the epithelium. This absorption is a consequence of the mucus displacement by the cilia along generations with an interface between the epithelium and the airway surface layer of decreasing area. The numerical results obtained are in good agreement with previously published experimental data, thus validating our approach. We also briefly discuss how our results can improve the understanding and, possibly, the treatment of pulmonary diseases.


Gas chromatography-electron ionization and chemical ionization mass spectrometric analysis of serum mexiletine concentration after derivatization with 2,2,2-trichloroethyl chloroformate: a novel derivative.
Mexiletine (Mexitil) is an antiarrhythmic agent used in the treatment of ventricular arrhythmia. The drug has a narrow therapeutic window, and monitoring its serum concentration is recommended. The authors describe a gas chromatography-mass spectrometric (GC/MS) assay of mexiletine using selected ion monitoring. Mexiletine was extracted from alkaline serum with dichloromethane followed by derivatization with 2,2,2-trichloroethyl chloroformate. The reaction was completed in 30 minutes at 70 degrees C. N-propylamphetamine was used as the internal standard. The ions monitored were m/z 58, 102, 122, 232, and 234 for derivatized mexiletine and m/z 56, 91, 131, 260, and 262 for the derivatized internal standard. The within-run precision at a serum mexiletine concentration of 1 mg/l was 1.7% (mean = 0.981, SD = 0.017 mg/l, n = 8) and the between-run precision was 3.3% (mean = 0.983, SD = 0.033 mg/l, n = 6). The assay was linear for serum mexiletine concentrations of 0.2 to 2.5 mg/l. The detection limit was 0.1 mg/l. The authors observed no carry-over problem in their assay. They observed a good correlation between mexiletine concentrations measured by a reference laboratory (Associated Regional University Pathologists, Salt Lake City, UT, U.S.A.) and by the new GC/MS assay.


Recent advances in hepatitis C virus treatment: review of HCV protease inhibitor clinical trials.
Hepatitis C virus (HCV) infection affects millions of people world-wide, and chronic infection can result in end-stage liver disease and hepatocellular carcinoma. Conventional therapy to date has involved combination antiviral therapy including alpha-interferon and ribavirin; response rates with these drugs are variable based on both viral and host factors, such as HCV viral load, HCV genotype, HIV co-infection, host genetic polymorphisms (such as those in the IL28B region), and other factors. Recent advances in HCV treatment have included pegylated forms of alpha-interferon and, more recently, the development of specifically targeted antiviral therapy for HCV (STAT-C) with novel HCV protease inhibitors (PIs) for genotype 1 HCV. Although unlikely to be administered as monotherapy due to the potential for development of HCV PI drug resistance mutations, results of phase II trials of two PIs in development have recently been reported, demonstrating promising therapeutic efficacy of HCV PIs in combination with established conventional treatment. This review outlines the advances and the challenges in the development of these HCV PIs as effective HCV antiviral agents and their role in clinical practice.


How Notifications Affect Engagement With a Behavior Change App: Results From a Micro-Randomized Trial.
Drink Less is a behavior change app to help higher-risk drinkers in the United Kingdom reduce their alcohol consumption. The app includes a daily notification asking users to "Please complete your drinks and mood diary," yet we did not understand the causal effect of the notification on engagement nor how to improve this component of Drink Less. We developed a new bank of 30 new messages to increase users' reflective motivation to engage with Drink Less. This study aimed to determine how standard and new notifications affect engagement. Our objective was to estimate the causal effect of the notification on near-term engagement, to explore whether this effect changed over time, and to create an evidence base to further inform the optimization of the notification policy. We conducted a micro-randomized trial (MRT) with 2 additional parallel arms. Inclusion criteria were Drink Less users who consented to participate in the trial, self-reported a baseline Alcohol Use Disorders Identification Test score of ≥8, resided in the United Kingdom, were aged ≥18 years, and reported interest in drinking less alcohol. Our MRT randomized 350 new users to test whether receiving a notification, compared with receiving no notification, increased the probability of opening the app in the subsequent hour, over the first 30 days since downloading Drink Less. Each day at 8 PM, users were randomized with a 30% probability of receiving the standard message, a 30% probability of receiving a new message, or a 40% probability of receiving no message. We additionally explored time to disengagement, with the allocation of 60% of eligible users randomized to the MRT (n=350) and 40% of eligible users randomized in equal number to the 2 parallel arms, either receiving the no notification policy (n=98) or the standard notification policy (n=121). Ancillary analyses explored effect moderation by recent states of habituation and engagement. Receiving a notification, compared with not receiving a notification, increased the probability of opening the app in the next hour by 3.5-fold (95% CI 2.91-4.25). Both types of messages were similarly effective. The effect of the notification did not change significantly over time. A user being in a state of already engaged lowered the new notification effect by 0.80 (95% CI 0.55-1.16), although not significantly. Across the 3 arms, time to disengagement was not significantly different. We found a strong near-term effect of engagement on the notification, but no overall difference in time to disengagement between users receiving the standard fixed notification, no notification at all, or the random sequence of notifications within the MRT. The strong near-term effect of the notification presents an opportunity to target notifications to increase "in-the-moment" engagement. Further optimization is required to improve the long-term engagement. RR2-10.2196/18690.


Early diagnosis of oral cancer using a hybrid arrangement of deep belief networkand combined group teaching algorithm.
Oral cancer can occur in different parts of the mouth, including the lips, palate, gums, and inside the cheeks. If not treated in time, it can be life-threatening. Incidentally, using CAD-based diagnosis systems can be so helpful for early detection of this disease and curing it. In this study, a new deep learning-based methodology has been proposed for optimal oral cancer diagnosis from the images. In this method, after some preprocessing steps, a new deep belief network (DBN) has been proposed as the main part of the diagnosis system. The main contribution of the proposed DBN is its combination with a developed version of a metaheuristic technique, known as the Combined Group Teaching Optimization algorithm to provide an efficient system of diagnosis. The presented method is then implemented in the "Oral Cancer (Lips and Tongue) images dataset" and a comparison is done between the results and other methods, including ANN, Bayesian, CNN, GSO-NN, and End-to-End NN to show the efficacy of the techniques. The results showed that the DBN-CGTO method achieved a precision rate of 97.71%, sensitivity rate of 92.37%, the Matthews Correlation Coefficient of 94.65%, and 94.65% F1 score, which signifies its ability as the highest efficiency among the others to accurately classify positive samples while remaining the independent correct classification of negative samples.


Childhood multifocal skeletal non-Hodgkin lymphoma is a differential diagnosis of battered child syndrome.
Skeletal lesions observed in battered children include fractures, subperiostal new bone formation, irregular metaphyses, and cone-shaped epiphyses. A 4-year-old girl presented with bone pain. Radiographs revealed multiple fractures and epiphyseal and metaphyseal lesions highly significant for battered child syndrome. Six weeks later the authors diagnosed primary multifocal skeletal non-Hodgkin lymphoma. This case report illustrates that multifocal skeletal non-Hodgkin lymphoma is a differential diagnosis of battered child syndrome.


Lessons from the evaluation of the UK's NHS R&D implementation methods programme.
Concern about the effective use of research was a major factor behind the creation of the NHS R&D Programme in 1991. In 1994, an advisory group was established to identify research priorities in research implementation. The Implementation Methods Programme (IMP) flowed from this, and its commissioning group funded 36 projects. In 2000 responsibility for the programme passed to the National Co-ordinating Centre for NHS Service Delivery and Organisation R&D, which asked the Health Economics Research Group (HERG), Brunel University, to conduct an evaluation in 2002. By then most projects had been completed. This evaluation was intended to cover: the quality of outputs, lessons to be learnt about the communication strategy and the commissioning process, and the benefits from the projects. We adopted a wide range of quantitative and qualitative methods. They included: documentary analysis, interviews with key actors, questionnaires to the funded lead researchers, questionnaires to potential users, and desk analysis. Quantitative assessment of outputs and dissemination revealed that the IMP funded useful research projects, some of which had considerable impact against the various categories in the HERG payback model, such as publications, further research, research training, impact on health policy, and clinical practice. Qualitative findings from interviews with advisory and commissioning group members indicated that when the IMP was established, implementation research was a relatively unexplored field. This was reflected in the understanding brought to their roles by members of the advisory and commissioning groups, in the way priorities for research were chosen and developed, and in how the research projects were commissioned. The ideological and methodological debates associated with these decisions have continued among those working in this field. The need for an effective communication strategy for the programme as a whole was particularly important. However, such a strategy was never developed, making it difficult to establish the general influence of the IMP as a programme. Our findings about the impact of the work funded, and the difficulties faced by those developing the IMP, have implications for the development of strategic programmes of research in general, as well as for the development of more effective research in this field.


Clinical features and genetic analysis of a case series of skeletal ciliopathies in a prenatal setting.
Short-rib polydactyly syndrome (SRPS) refers to a group of lethal skeletal dysplasias that can be difficult to differentiate between subtypes or from other non-lethal skeletal dysplasias such as Ellis-van Creveld syndrome and Jeune syndrome in a prenatal setting. We report the ultrasound and genetic findings of four unrelated fetuses with skeletal dysplasias. Systemic prenatal ultrasound examination was performed in the second or third trimester. Genetic tests including GTG-banding, single nucleotide polymorphism (SNP) array and exome sequencing were performed with amniocytes or aborted fetal tissues. The major and common ultrasound anomalies for the four unrelated fetuses included short long bones of the limbs and narrow thorax. No chromosomal abnormalities and pathogenic copy number variations were detected. Exome sequencing revealed three novel variants in the DYNC2H1 gene, namely NM_001080463.2:c.6809G > A p.(Arg2270Gln), NM_001080463.2:3133C > T p.(Gln1045Ter), and NM_001080463.2:c.337C > T p.(Arg113Trp); one novel variant in the IFT172 gene, NM_015662.3:4540-5 T > A; and one novel variant in the WDR19 gene, NM_025132.4:c.2596G > C p.(Gly866Arg). The genotypes of DYNC2H1, IFT172 and WDR19 and the phenotypes of the fetuses give hints for the diagnosis of short-rib thoracic dysplasia (SRTD) with or without polydactyly 3, 10, and 5, respectively. Our findings expand the mutation spectrum of DYNC2H1, IFT172 and WDR19 associated with skeletal ciliopathies, and provide useful information for prenatal diagnosis and genetic counseling on rare skeletal disorders.


Patient-reported outcomes after hepatic resection of colorectal cancer metastases.
Hepatic resection of colorectal carcinoma (CRC) liver metastases is increasing, but evidence for the impact of surgery on patient-reported outcomes (PROs) is limited. This study aimed to describe comprehensively the impact of liver surgery for CRC hepatic metastases on PROs. Consecutive patients selected for hepatic resection completed the European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire-C30 and Quality of Life Questionnaire-Liver Metastases C21 before and 3, 6, and 12 months after surgery. For functional scales, mean scores with 95% CIs were calculated at each time point, with differences in scores of at least 10 points considered clinically significant. Responses to symptom scales and items were categorized as minimal or severe. Proportions and 95% CIs for each symptom category were calculated. Hepatic surgery was planned in 241 patients but abandoned in nine because of unresectable disease. There were two postoperative deaths, 58 complications (25.2%), and 32 patients (14.9%) with disease recurrence. Questionnaire compliance was excellent (> 95% at all time points). After surgery, most functional aspects of health decreased, and the proportions of patients with severe symptoms increased; role function deteriorated significantly, and 30% of patients reported severe activity/vigor problems. Functional scales recovered by 6 months and were maintained at 1 year. Postoperative symptoms returned to baseline levels at 12 months, but 32.1% of patients reported severe problems with sexual dysfunction and 11.9% with abdominal pain. These findings provide new evidence regarding outcomes of liver resection for CRC metastases. It is recommended that patients be reassured that surgery has a minimal and short-lived detrimental impact on health.


Tunability enhancement of Yb:YLF based laser.
We present a comprehensive characterization of a laser, based on a high-quality heavily doped Yb:YLF crystal. The analysis includes the comparison of the laser efficiency in respect of the absorbed pump for both pi and sigma-polarization. In particular, for the tunable laser, we report an enhancement of the tuning range obtained by adapting the output coupler transmission in respect of the lasing wavelength. The experimental results obtained for various output coupling have also been compared with a numerical model in order to give information for a further enlargement of the tuning range.


The value of a nationwide vascular registry in understanding contemporary time trends of abdominal aortic aneurysm repair.
Nationwide vascular registries offer rapid feed-back in an environment of fast new technical development, as is the case with the treatment of abdominal aortic aneurysm (AAA). Furthermore, they offer an opportunity to study non-selected, population-based data. The aim of this review was to analyze time-trends in published papers from nationwide registries on AAA-repair. In contrast to several US reports, an increased rate of intact AAA repair, associated with the introduction of endovascular repair, was reported in a recent publication based on the Swedish Vascular Registry (Swedvasc). The rate of ruptured abdominal aortic aneurysm (rAAA) repair is stable in most reports, while some report a decreasing incidence. Most nationwide studies report a reducing mortality over time after intact AAA repair, while time trends on the mortality after ruptured AAA repair are more heterogenic.


Small-molecule ambipolar transistors.
Ambipolar transistor properties have been observed in various small-molecule materials. Since a small energy gap is necessary, many types of molecular designs including extended π-skeletons as well as the incorporation of donor and acceptor units have been attempted. In addition to the energy levels, an inert passivation layer is important to observe ambipolar transistor properties. Ambipolar transport has been observed in extraordinary π-electron systems such as antiaromatic compounds, biradicals, radicals, metal complexes, and hydrogen-bonded materials. Several donor/acceptor cocrystals show ambipolar transport as well.


Significance of alanine aminopeptidase N (APN) in bile in the diagnosis of acute cellular rejection after liver transplantation.
Allograft dysfunction after liver transplantation requires histopathologic examination for confirmation of the diagnosis, however, the procedure is invasive and its interpretation is not always accurate. The aim of this study was to find novel protein markers in bile for the diagnosis of acute cellular rejection (ACR) after liver transplantation. Quantitative proteomic analysis using the (18)O labeling method was used to search for bile proteins of interest. Nine recipients were selected who had liver dysfunction, diagnosed by liver biopsy, either with ACR (ACR group, n = 5) or without (LD group, n = 4). Donor bile samples were obtained from nine independent live liver donors. Enzyme activity in bile samples was assayed and liver biopsy specimens were immunostained for candidate protein of ACR. The analysis identified 78 proteins, among which alanine aminopeptidase N (APN/CD13) was considered a candidate marker of ACR. Comparative analysis of the ACR and LD groups showed high APN enzyme activity in three (60%) of five cases of the ACR group, while it was as low as donor level in all patients of the LD group. APN enzyme activity in bile samples of liver dysfunction liver transplantation (LDLT) recipients of the ACR group collected within 3 d before biopsy-confirmed ACR (n = 10) was significantly higher (584 ± 434 U/g protein) than in those of recipients free of ACR (n = 96, 301 ± 271 U/g protein) (P = 0.004). APN overexpression along bile canaliculi was observed during ACR in all five cases of the ACR group. APN in bile seems to be a useful and noninvasive biomarker of ACR after liver transplantation.


Temperature dependence of ethanol depression in mice.
The relationship between environmental temperature, body temperature and ethanol depression was investigated in male C57BL/6J mice. Animals were injected with ethanol (3.6 g/kg) or saline and placed in an experimental or normal room temperature (control) environment. Environmental temperatures from 12-37 degrees C significantly influenced rectal temperature, sleep-time and wake-up brain ethanol concentrations. The rectal temperatures of ethanol-treated animals ranged from 1.5 degrees C greater to 11.3 degrees C lower than their matched saline controls, supporting indications that ethanol impairs normal thermoregulation. Sleep-time decreased up to 43% and wake-up brain ethanol concentrations increased as rectal temperature decreased from approximately 38-32 degrees C. More severe hypothermia after ethanol (rectal temperature less than 29 degrees C) increased sleep-time and decreased wake-up brain ethanol concentrations. These results indicate that sleep-time and brain sensitivity to ethanol vary with rectal temperature in accordance with predictions based on membrane expansion and partition theories of anesthesia and the anticipated anesthetic-like actions of severe hypothermia.


Heavy ion irradiation inhibits in vitro angiogenesis even at sublethal dose.
Angiogenesis is essential for tumor growth and metastasis. Because endothelial cells are genetically stable, they rarely acquire resistance to anticancer modalities, and could, thus, be a suitable target for radiation therapy. Heavy ion radiation therapy has attracted attention as an effective modality for cancer therapy because of its highly lethal effects, but the effects of heavy ion irradiation on in vitro cell function associated with angiogenesis have not been reported. Our study found that in vitro angiogenesis was inhibited by high linear energy transfer carbon ion irradiation even at sublethal dose (0.1 Gy). ECV304 and HUVEC human umbilical vascular endothelial cells were irradiated with 290 MeV carbon ion beams of approximately 110 keV/ micro m or 4 MV X-ray of approximately 1 keV/ micro m. Their adhesiveness and migration to vitronectin or osteopontin were inhibited, and capillary-like tube structures in three-dimensional culture were destroyed after carbon ion irradiation concomitant with the inhibition of matrix metalloproteinase-2 activity, down-regulation of alphaVbeta3 integrin, which is one of the adhesion molecules, slight up-regulation of membrane type1- matrix metalloproteinase, and significant up-regulation of tissue inhibitor of metalloproteinase-2. On the other hand, sublethal X-ray irradiation promoted migration of endothelial cells, and the capillary-like tube structure in three-dimensional culture progressed even after 16 Gy irradiation. These results provide an implication that heavy ion beam therapy could be superior to conventional photon beam therapy in preventive effects on in vitro angiogenesis even at sublethal dose, and might inhibit angiogenesis in vivo.


Using light to see and control membrane traffic.
Cellular compartmentalization into discrete organelles is maintained by membrane trafficking including vesiculation and tubulation. Recent advances in superresolution imaging have begun to bring these small and dynamic events into focus. Most nanoscopes exploit, and are limited by, switching dyes ON and OFF. Using ground state depletion to switch dyes into long-lived dark states can exploit specific photophysical properties of dyes, such as redox potential or pK(a), and expand the repertoire of nanoscopy probes for multicolor imaging. Seeing is not enough, and new technologies based on homodimerization, heterodimerization and selective release can manipulate membrane trafficking in pulse-chase and light-controlled ways. Herein we highlight the utility and promise of these strategies and discuss their current limitations.


Optimal beamforming in ultrasound using the ideal observer.
Beamforming of received pulse-echo data generally involves the compression of signals from multiple channels within an aperture. This compression is irreversible, and therefore allows the possibility that information relevant for performing a diagnostic task is irretrievably lost. The purpose of this study was to evaluate information transfer in beamforming using a previously developed ideal observer model to quantify diagnostic information relevant to performing a task. We describe an elaborated statistical model of image formation for fixed-focus transmission and single-channel reception within a moving aperture, and we use this model on a panel of tasks related to breast sonography to evaluate receive-beamforming approaches that optimize the transfer of information. Under the assumption that acquisition noise is well described as an additive wide-band Gaussian white-noise process, we show that signal compression across receive-aperture channels after a 2-D matched-filtering operation results in no loss of diagnostic information. Across tasks, the matched-filter beamformer results in more information than standard delay-and-sum beamforming in the subsequent radio-frequency signal by a factor of two. We also show that for this matched filter, 68% of the information gain can be attributed to the phase of the matched-filter and 21% can be attributed to the amplitude. A 1-D matched filtering along axial lines shows no advantage over delay-andsum, suggesting an important role for incorporating correlations across different aperture windows in beamforming. We also show that a post-compression processing before the computation of an envelope is necessary to pass the diagnostic information in the beamformed radio-frequency signal to the final envelope image.