Academic Institutions and One Health: Building Capacity for Transdisciplinary Research Approaches to Address Complex Health Issues at the Animal-Human-Ecosystem Interface.
To improve health at the human, animal, and ecosystem interface, defined as One Health, training of researchers must transcend individual disciplines to develop a new process of collaboration. The transdisciplinary research approach integrates frameworks and methodologies beyond academic disciplines and includes involvement of and input from policy makers and members of the community. The authors argue that there should be a significant shift in academic institutions' research capacity to achieve the added value of a transdisciplinary approach for addressing One Health problems. This Perspective is a call to action for academic institutions to provide the foundations for this salient shift. The authors begin by describing the transdisciplinary approach, propose methods for building transdisciplinary research capacity, and highlight three value propositions that support the case. Examples are provided to illustrate how the transdisciplinary approach to research adds value through improved sustainability of impact, increased cost-effectiveness, and enhanced abilities to mitigate potentially harmful unintended consequences. The authors conclude with three key recommendations for academic institutions: (1) a focus on creating enabling environments for One Health and transdisciplinary research, (2) the development of novel funding structures for transdisciplinary research, and (3) training of "transmitters" using real-world-oriented educational programs that break down research silos through collaboration across disciplines.


Inhibitory effect of veterinary antibiotics on denitrification in groundwater: a microcosm approach.
Veterinary antibiotics in groundwater may affect natural microbial denitrification process. A microcosm study was conducted to evaluate the influence of sulfamethazine and chlortetracycline at different concentrations (0, 0.01, 0.1, and 1.0 mg/L) on nitrate reduction in groundwater under denitrifying condition. Decrease in nitrate removal and nitrite production was observed with the antibiotics. Maximum inhibition of nitrate removal was observed after seven days of incubation with 0.01 mg/L sulfamethazine (17.0%) and 1.0 mg/L chlortetracycline (15.4%). The nitrite production was inhibited with 1.0 mg/L sulfamethazine to 82.0% and chlortetracycline to 31.1%. The initial/final nitrate concentrations indicated that 0.01 mg/L sulfamethazine and 1.0 mg/L chlortetracycline were most effective in inhibiting activity of denitrifying bacteria in groundwater. After 12 days of incubation, the sulfamethazine biodegradation was observed whereas chlortetracycline was persistent. Sulfamethazine and chlortetracycline in groundwater could inhibit the growth and capability of naturally occurring denitrifying bacteria, thereby threatening nitrate pollution in groundwater.


Ageing populations and childhood infections: the potential impact on epidemic patterns and morbidity.
Population decline, arising from below replacement fertility and possibly giving rise to substantial changes in age distribution, is a feature of many industrialized developed countries; Italy is one of the most notable European examples. The potential influence of this phenomenon on prevalence of chronic non-infectious disease is well known, but little attention to date has been paid to the impact on severe disease due to childhood infections in those cases where control is insufficient to achieve elimination. A transmission dynamics model incorporating realistic demography is used to investigate the possible impact of population decline and ageing and suboptimal vaccination uptake on the age distribution of incidence of measles infection and of consequent mortality. Data from Italy is used to parameterize the model. Population ageing in the absence of vaccination is shown to reduce per capita incidence of infection but also to increase average and upper quartile ages at infection. The effect is substantially enhanced by significantly suboptimal vaccination uptake, when disease-induced mortality has, for a period, the potential to exceed that in the absence of vaccination. Although a substantially increased burden from chronic non-infectious disease has frequently been proposed as a consequence of population decline, there is also potential for an increase in morbidity and mortality from measles and other childhood infectious diseases, particularly where vaccine uptake is substantially below the optimum. Rubella is highlighted as a particular cause for concern. This work also has implications for less-developed countries.


[Pharmacodynamics and pharmacokinetics of the new specific bradycardic preparation bradizole].
The new drug Bradizol (SM-345), a 2-mercaptobenzimidazole derivative producing a specific bradycardic effect, is quite rapidly eliminated upon intravenous bolus in anaesthetiezed cats. After a combined injection (bolus followed by a 60-min infusion), a constant bradizol concentration in the blood plasma is observed over a time period of 90 min. The bradizol-induced bradycardic effect and the drug concentration in the blood plasma are tightly correlated. Upon per os administration of bradizol to the experimental animals, neither the drug is detected in the blood plasma, nor the bradycardic effect is observed. It is suggested that bradizol, possessing antiarrhythmic, antifibrillator, and antiischemic properties, can be used (by intravenous injections) for the therapy of patients with diverse tachyarrhythmia geneses, including those with the ischemic heart disease.


Matters of fact and politics: Generating expectations of cancer screening.
Many countries, including Australia, the United Kingdom and the United States have established national screening programs in the effort to advance the early diagnosis of cancers. Australia has population screening programs for breast, bowel and cervical cancers, and this article focuses on breast and cervical cancer screening as the two longest running programs in Australia. While these screening programs are well-established and report relatively high participation rates, the effectiveness of population screening is a contested issue, subject to significant, ongoing debate about its purported benefits (Armstrong, 2019). In this article, we draw on ideas from sociology of science on the construction of scientific facts to analyse how evidentiary claims are presented in policy documents for Australia's breast and cervical cancer screening programs, and the implications for those who are the targets of screening. We explore how screening-related information assumes the status of scientific 'facts', and argue that presenting information as neutral and objective obscures the political choices involved in its generation. Importantly, some of the claims presented in the policy documents have a tendency to emphasise the benefits, and minimise the risks and harms of population-based screening. In doing so, we suggest that the current national policies may be contributing to sustaining expectations of screening that are higher than warranted. Higher expectations may bring with them unintended societal and economic costs to the public. We conclude by noting how deeply ingrained socio-cultural meanings of cancer shape public expectations of the protective value of screening, which allows current screening approaches to become further entrenched and resistant to challenge.


Beneficial effect of oral tigecycline treatment on Clostridium difficile infection in gnotobiotic piglets.
The efficacy of oral tigecycline treatment (2 mg/kg of body weight for 7 days) of Clostridium difficile infection (CDI) was evaluated in the gnotobiotic pig model, and its effect on human gut microflora transplanted into the gnotobiotic pig was determined. Tigecycline oral treatment improved survival, clinical signs, and lesion severity and markedly decreased concentrations of Firmicutes but did not promote CDI. Our data showed that oral tigecycline treatment has a potential beneficial effect on the treatment of CDI.


Selectivity, sharing and competitive interactions in the regulation of Hoxb genes.
The clustered organisation of Hox complexes is highly conserved in vertebrates and the reasons for this are believed to be linked with the regulatory mechanisms governing their expression. In analysis of the Hoxb4-Hoxb6 region of the HoxB complex we identified enhancers which lie in the intergenic region between Hoxb4 and Hoxb5, and which are capable of mediating the correct boundaries of neural and mesodermal expression for Hoxb5. We examined their regulatory properties in the context of the local genomic region spanning the two genes by transgenic analysis, in which each promoter was independently marked with a different reporter, to monitor simultaneously the relative transcriptional read-outs from each gene. Our analysis revealed that within this intergenic region: (i) a limb and a neural enhancer selectively activate Hoxb4 as opposed to Hoxb5; (ii) a separate neural enhancer is able to activate both genes, but expression is dependent upon competition between the two promoters for the enhancer and is influenced by the local genomic context; (iii) mesodermal enhancer activities can be shared between the genes. We found similar types of regulatory interactions between Hoxb5 and Hoxb6. Together these results provide evidence for three separate general mechanisms: selectivity, competition and sharing, that control the balance of cis-regulatory interactions necessary for generating the proper spatial and temporal patterns of Hox gene expression. We suggest that these mechanisms are part of a regulatory basis for maintenance of Hox organisation.


Mutagenic and inhibitory effects of ribavirin on hepatitis C virus RNA polymerase.
Crotty et al. recently proposed the primary antiviral action of ribavirin to be that of a potent RNA mutagen [Crotty, S., Maag, D., Arnold, J. J., Zhong, W., Lau, J. Y., Hong, Z., Andino, R., and Cameron, C. E. (2000) Nat. Med. 6, 1375-1379]. Here we investigate the effect of ribavirin triphosphate (RTP) on RNA synthesis catalyzed by a full-length hepatitis C virus (HCV) RNA polymerase in vitro. HCV polymerase can use RTP as a nucleotide substrate in a template-dependent manner, incorporating it opposite a pyrimidine (C or U) template residue, but not a purine (A or G). Kinetic analysis revealed that incorporation of ribavirin monophosphate (RMP) across from C is 3 times more efficient catalytically than that across from U, as determined by the k(cat)/K(m) parameter. The efficiency of RMP incorporation, however, is 50-100 fold lower than that of the natural NMP. RMP incorporation does not lead to termination of RNA chain synthesis, as evidenced by the ability of the polymerase to extend its RNA product many nucleotides beyond the site of RMP incorporation. However, multiple-RMP incorporation at low GTP concentrations induced the formation of stalled elongation complexes, particularly at the template region containing consecutive C residues. Most, but not all, such elongation blocks can be relieved by the re-addition of GTP. When ribavirin is present in the RNA template, pyrimidine (but neither purine nor ribavirin) monophosphate is incorporated opposite ribavirin, but at an exceedingly low catalytic efficiency (200-3000-fold lower) compared to the efficiencies of those templated by A or G. Consequently, the level of RNA synthesis on a ribavirin-containing template is significantly reduced. These findings suggest that ribavirin not only is mutagenic but also interferes with HCV polymerase-mediated RNA synthesis.


Potential ocular and systemic COVID-19 prophylaxis approaches for healthcare professionals.
The COVID-19 pandemic has brought with it, innumerable challenges in healthcare, both through the direct burden of morbidity and mortality of the disease, and also by the curtailing of other essential albeit less emergency medical services to reduce the risk of community spread. Reports from around the world are showing mounting number of cases even in healthcare professionals spite of usage of adequate personal protective equipment. There are a number of factors which could account for this, be it the affinity of the virus to the respiratory and other mucosa or to patient risk factors for developing severe forms of the disease. In view of the growing need for resuming other medical services, it is essential to find newer ways to protect ourselves better, whether by systemic or topical mucosal prophylaxis with various medications or lifestyle changes promoting wellbeing and immunity. This article discusses additional prophylactic measures including drug repurposing or new indication paradigms to render protection. Certain medications such as chloroquine, trehalose, antihistaminics, and interferons used topically for various ocular conditions with reasonably good safety records are known to have anti-viral properties. Hence, can be harnessed in preventing SARS-CoV-2 attachment, entry, and/or replication in host cells. Similarly, use of hypertonic saline for nasal and oral mucosa and dietary changes are possible methods of improving our resistance. These additional prophylactic measures can be cautiously explored by healthcare professionals to protect themselves and their patients.


Experiences from a multimodal rhythm and music-based rehabilitation program in late phase of stroke recovery - A qualitative study.
Rehabilitative stroke interventions based on principles of multimodal stimulation have the potential to profoundly affect neuroplastic processes beyond the sub-acute phase. In order to identify important core mechanisms, there is a need to explore how interventions that combine physical, social, sensory, and cognitively challenging activities are perceived and experienced by the participants. This qualitative study, based on an interpretive interactionist perspective, explored the experiences of stroke survivors who participated in a group-based multimodal rehabilitation program based on rhythm and music. Within four weeks after completion of the multimodal rehabilitation program, face-to-face semi-structured interviews were conducted on a single occasion with 15 purposively selected individuals (mean age 65, 8 men, 7 women). The interview duration was between 13 and 44 minutes. Qualitative content analysis with an inductive approach was used to analyze data. Three categories were identified, each containing several sub-categories: To be intellectually challenged (energy-consuming activity and coordinating multiple input and output), Perceived therapeutic benefits (motor skills, cognitive skills, emotional and psychological responses), and Pros and cons with social integration (fellowship, competing with others, and instructor characteristics). From these categories, an overall theme was derived: The multifaceted layers of multimodal stimulation. Enjoying music, being part of a group with peers, a skilled instructor, and being able to manage the challenging movements, were related to positive experiences. In contrast, negative experiences were associated with not being able to perform the exercises, and with group members who dominated the conversational space. This study shows that access to a multimodal rehabilitation program with rhythm and music as operating ingredients may contribute to positive experiences for many individuals in a late phase after stroke in terms of motor, cognitive, as well as emotional enhancements. Important components were the music, the social interaction, the challenging exercises, and the skilled instructor.


In vivo characterization of vasocontractile activities in erythrocytes.
Partially purified protein from washed and artificially hemolyzed erythrocytes, known to cause significant contractions of isolated canine cerebral vessels in vitro, was injected into the cisterna magna of intact anesthetized dogs. Cerebral blood flow, measured by the xenon-133 washout technique, decreased from a control value of 49.5 +/- 1.17 ml/100 gm/min to an experimental value of 34.1 +/- 1.65 ml/100 gm/min at 2 hours. Cerebral vascular resistance rose from a control value of 2.05 +/- 0.17 PRU (peripheral resistance units) to an experimental value of 2.91 +/- 0.25 PRU at 2 hours. Mean arterial blood pressure, heart rate, intracranial pressure, and cerebral perfusion pressure remained stable. Cardiac output also fell significantly (in 2-hour control animals it was 2.89 +/- 0.37 liter/min, and in 2-hour experimental animals 1.43 +/- 0.13 liter/min) and peripheral vascular resistance rose. These changes were evident by 10 minutes after the cisternal injection of the hemolysate protein, and remained for the duration of the 2-hour monitoring period. Serial vertebrobasilar angiograms demonstrated marked narrowing of the intracranial basilar artery when compared to control values. The narrowing persisted for several days in most animals, and tended to increase with time. Relaxation occurred by the 10th through the 14th day. The authors conclude that this experimental preparation may be a useful model for both in vitro and in vivo investigation of chronic cerebral vasospasm.


Head-mounted display augmented reality to guide pedicle screw placement utilizing computed tomography.
Augmented reality has potential to enhance surgical navigation and visualization. We determined whether head-mounted display augmented reality (HMD-AR) with superimposed computed tomography (CT) data could allow the wearer to percutaneously guide pedicle screw placement in an opaque lumbar model with no real-time fluoroscopic guidance. CT imaging was obtained of a phantom composed of L1-L3 Sawbones vertebrae in opaque silicone. Preprocedural planning was performed by creating virtual trajectories of appropriate angle and depth for ideal approach into the pedicle, and these data were integrated into the Microsoft HoloLens using the Novarad OpenSight application allowing the user to view the virtual trajectory guides and CT images superimposed on the phantom in two and three dimensions. Spinal needles were inserted following the virtual trajectories to the point of contact with bone. Repeat CT revealed actual needle trajectory, allowing comparison with the ideal preprocedural paths. Registration of AR to phantom showed a roughly circular deviation with maximum average radius of 2.5 mm. Users took an average of 200 s to place a needle. Extrapolation of needle trajectory into the pedicle showed that of 36 needles placed, 35 (97%) would have remained within the pedicles. Needles placed approximated a mean distance of 4.69 mm in the mediolateral direction and 4.48 mm in the craniocaudal direction from pedicle bone edge. To our knowledge, this is the first peer-reviewed report and evaluation of HMD-AR with superimposed 3D guidance utilizing CT for spinal pedicle guide placement for the purpose of cannulation without the use of fluoroscopy.


Carbonic anhydrase C in white-skeletal-muscle tissue.
We investigated the activity of carbonic anhydrase in blood-free perfused white skeletal muscles of the rabbit. Carbonic anhydrase activities were measured in supernatants and in Triton extracts of the particulate fractions of white-skeletal-muscle homogenate by using a rapid-reaction stopped-flow apparatus equipped with a pH electrode. An average carbonic anhydrase concentration of about 0.5 microM was determined for white skeletal muscle. This concentration is about 1% of that inside the erythrocyte. Some 85% of the muscle enzyme was found in the homogenate supernatant, and only 15% appeared to be associated with membranes and organelles. White-skeletal-muscle carbonic anhydrase was characterized in terms of its Michaelis constant and catalytic-centre activity (turnover number) for CO2 and its inhibition constant towards ethoxzolamide. These properties were identical with those of the rabbit erythrocyte carbonic anhydrase C, suggesting that a type-C enzyme is present in white skeletal muscle. Affinity chromatography of muscle supernatant and of lysed erythrocytes showed that, whereas rabbit erythrocytes contain about equal amounts of carbonic anhydrase isoenzymes B and C, the B isoenzyme is practically absent from white skeletal muscle. Similarly, ethoxzolamide-inhibition curves suggested that white skeletal muscle contains no carbonic anhydrase A. It is concluded that white skeletal muscle contains essentially one carbonic anhydrase isoenzyme, the C form, most of which is probably of cytosolic origin.


Polyclonal antibodies to glutathione S-transferase--verotoxin subunit a fusion proteins neutralize verotoxins.
The A1 subunits of verotoxin-1 (VT1) and VT2 genes were cloned into pGEX-4T-2 for the expression of glutathione S-transferase (GST) fusion proteins. The N-terminal and the transmembrane regions of the A1 subunits were excluded from the constructs in order to increase the product yields. Polyclonal anti-VT1A1 and anti-VT2A1 antibodies were produced by immunizing rabbits with GST-VT1A1 and GST-VT2A1 fusion proteins, respectively. The antibodies were tested for their ability to neutralize active toxins from 45 VT-producing Escherichia coli (VTEC) strains. The antibodies had significantly high neutralizing activities against their homologous toxins. The average percentages of neutralization of VT1 by anti-GST-VT1A1 and anti-GST-VT2A1 were 76.7% +/- 7.9% and 3.6% +/- 2.3%, respectively, and those of VT2 were 1.7% +/- 2.3% and 82.5% +/- 13.9%, respectively. VT2 variant toxin was neutralized by anti-GST-VT2A1, with cross neutralization being a possible consequence of sequence homology between VT2 and a VT2 variant. To our knowledge, this is the first report on the production of polyclonal antibodies from GST-VT fusion proteins. The antibodies were shown to exhibit specific toxin neutralizing activities and may be useful for immunological diagnosis of VTEC infections.


Wafer-scale high-throughput ordered growth of vertically aligned ZnO nanowire arrays.
This article presents an effective approach for patterned growth of vertically aligned ZnO nanowire (NW) arrays with high throughput and low cost at wafer scale without using cleanroom technology. Periodic hole patterns are generated using laser interference lithography on substrates coated with the photoresist SU-8. ZnO NWs are selectively grown through the holes via a low-temperature hydrothermal method without using a catalyst and with a superior control over orientation, location/density, and as-synthesized morphology. The development of textured ZnO seed layers for replacing single crystalline GaN and ZnO substrates extends the large-scale fabrication of vertically aligned ZnO NW arrays on substrates of other materials, such as polymers, Si, and glass. This combined approach demonstrates a novel method of manufacturing large-scale patterned one-dimensional nanostructures on various substrates for applications in energy harvesting, sensing, optoelectronics, and electronic devices.


Differential activation of psoas major and rectus femoris during active straight leg raise to end range.
The purpose of this study was to investigate the activation of the hip flexor and abdominal muscles during an active straight leg raise (ASLR) to end range of hip flexion. Data were recorded from nine healthy men. Fine-wire electromyography (EMG) electrodes were inserted into psoas major (PM), and surface electrodes were placed over rectus femoris (RF), rectus abdominis, obliquus externus abdominis (OE), and obliquus internus abdominis/transversus abdominis (OI/TrA). EMG and kinematic data were obtained during concentric, hold (at end range) and eccentric phases of an ASLR. Concentric and eccentric movements were divided into three phases (early, mid, and late). Onsets of EMG relative to the onset of the ALSR movement and EMG amplitudes in each phase were compared between muscles. Onsets of the PM (-33 ± 245 ms) and RF (-3 ± 119 ms) EMG prior to leg elevation were significantly earlier than those of the OE and OI/TrA. PM EMG showed highest activation in the late concentric, hold, early eccentric phase, and was significantly higher than RF EMG. OI/TrA EMG was significantly greater in mid and late concentric, hold, and early eccentric phase than other phases. During the ASLR, unlike RF, PM EMG continues to increase towards the end range of hip flexion. Activation of OI/TrA muscle may be involved in control trunk and pelvic movement.


[Epidermoid epitheliomas of the upper respiratory and digestive pathways in females].
Ninety-five females with squamous cell carcinoma of the head and neck are reported. Tobacco and alcohol habits were present in only 18% of patients; in these cases tumoral behavior and prognosis are quite similar to those of male patients. When no particular epidemiology was found, female patients present special tumoral localizations as gum, cheek or palate. In addition these patients are elderly, mean age above 75 years. Radiotherapeutic management of female patients allow the same results than in treating males; consequently, since female patients are more elderly than males, these therapeutic results advocate aggressive therapy even in female patients above 80 years old.


Computerized binary scale of auditory speech hallucinations (cbSASH).
Evidence indicates that the neuropathology of auditory verbal hallucinations (AVH) varies according to their phenomenological characteristics. Therefore, AVH should be subgrouped accordingly in hallucinations research. As evaluation of these characteristics depends entirely on the patient report, obtaining measurement of the reliability of these reports is crucial. A computerized binary scale of auditory speech hallucinations (cbSASH) was developed to evaluate the phenomenology of AVH. It includes two subscales (inconsistency and malingering) to assess the reliability of the patient report. The cbSASH was administered along with MMPI-2, a general psychopathology scale, which includes similar validity subscales. Thirty-four psychotic patients with history of AVH were enrolled in this study. The scores on the inconsistency and malingering subscales of the cbSASH were correlated with the scores on the corresponding validity subscales in the MMPI-2. The combination of the malingering and inconsistency subscales provided robust measures of the reliability and ability of the patients' descriptions of their hallucinations. The cbSASH provides a reliable and comprehensive evaluation of the phenomenology of AVH. Consequently, it is possible to subgroup patients according to the characteristics of their hallucinations. This refinement of AVH phenotypes could reduce the noise and inconsistency noted in AVH and psychosis research.


A Conceptual Framework for Use of Increased Endurance Time During Constant Work Rate Cycle Ergometry as a Patient-Focused Meaningful Outcome in COPD Clinical Trials.
The Chronic Lung Disease Biomarker and Clinical Outcome Assessment Qualification Consortium (CBQC) evaluates the potential of biomarkers and outcome measures as drug development tools. Exercise endurance is an objective indicator of treatment benefit, closely related to daily physical function. Therefore, it is an ideal candidate for an outcome for drug development trials. Unfortunately, no exercise endurance measure is qualified by regulatory authorities for use in trials of chronic obstructive pulmonary disease (COPD) and no approved COPD therapies have claims of improving exercise endurance. Consequently, it has been challenging for developers to consider this outcome when designing clinical trials for new therapies. Endurance time during constant work rate cycle ergometry (CWRCE), performed on an electronically braked stationary cycle ergometer, provides an exercise endurance measure under standardized conditions. Baseline individualized work rate for each participant is set using an incremental test. During CWRCE the patient is encouraged to continue exercising for as long as possible. Although not required, physiological and sensory responses (e.g., pulmonary ventilation, heart rate, dyspnea ratings) are frequently collected to support interpretation of endurance time changes. Exercise tolerance limit is reached when the individual is limited by symptoms, unable to maintain pedaling cadence or unable to continue safely. At exercise cessation, exercise duration is recorded. An CWRCE endurance time increase from the pre-treatment baseline is proposed as a key efficacy endpoint in clinical trials. In COPD, improved exercise endurance has a direct relationship to the experience of physical functioning in daily life, which is a patient-centered, meaningful benefit.


Ezrin-Radixin-Moesin Binding Phosphoprotein 50: A Potential Novel Biomarker in Human Papilloma Virus-Associated Head and Neck Squamous Cell Carcinomas.
High-risk human papilloma virus (HR-HPV) has increasingly been associated with head and neck squamous cell carcinoma (HNSCC), in particular oropharyngeal cancers. Ezrin-Radixin-Moesin Binding Phosphoprotein 50 (EBP50), a putative tumour suppressor, localises to the plasma membrane in suprabasal epithelium and to the cytoplasm in proliferative basal layers, and is a target for degradation by the HR-HPV E6 oncoprotein. The aim of this study was to investigate EBP50 protein expression patterns in HNSCC in a large Scottish cohort to determine if there was a correlation with HPV status and clinical outcomes. EBP50 expression patterns were assessed in 156 HNSCC including oropharyngeal (37.8%), laryngeal (24%), oral (19%) and other sites (18.5%), which were genotyped for presence of HR-HPV. HNSCC were generally negative for membranous EBP50. EBP50 expression was either cytoplasmic/absent, being 'predominantly cytoplasmic' in 76 (49%), 'weak/negligible cytoplasmic' in 44 (28%), 'strongly cytoplasmic' in 5 (3%), 'heterogeneous' in 26 (17%) and 'other' in 5 (3%) samples. Forty tumours (25%) were positive for HPV DNA, predominantly HR-HPV 16, and 44 (28%) were p16 positive. The majority of tumours (71%) with 'weak/negligible cytoplasmic' EBP50 expression originated in the oropharynx were more likely to have positive neck nodes, overexpression of p16 and positive tumour HR-HPV status (P < 0.001). Differences in EBP50 levels between oropharyngeal and non-oropharyngeal tumours may be linked to degradation of EBP50 by HR-HPV, and loss of EBP50 may therefore be a surrogate biomarker for HR-HPV infection in oropharyngeal tumours.


DNA polymerase delta, an essential enzyme for DNA transactions.
Many DNA transactions, such as replication, repair and recombination involve DNA synthesis and consequently require the action of DNA synthesizing enzymes called DNA polymerases (Pol). Eukaryotic cells contain at least six different Pols, named alpha, beta, gamma, delta, epsilon, and zeta. Among them Pol delta occupies important roles in DNA replication, nucleotide excision repair, base excision repair and VDJ recombination. Pol a has been extremely conserved in evolution from yeast to man. The function of Pol delta must be considered in the context of two other factors, called proliferating cell nuclear antigen and replication factor C, two protein complexes that build together the moving platform for Pol delta. This moving platform provides an important framework for dynamic properties of an accurate Pol delta such as its recruitment when its function is needed, the facilitation of Pol delta binding to the primer terminus, the increase in Pol delta processivity, the prevention of non-productive binding of the Pol delta to single-stranded DNA, the release of Pol delta after DNA synthesis and the bridging of Pol delta interactions to other replication proteins. In this review we summarize the current knowledge of Pol delta and will focus in particular to its structural conservation, its functional tasks in the cell and its interactions with other proteins.


Personality traits as predictors of OCD remission: A longitudinal study.
Personality traits may confer vulnerability to psychopathology. However, few studies have examined the association between personality traits and obsessive-compulsive disorder (OCD) course. The present study investigates personality traits, OCD symptom severity, and illness duration as a predictor of OCD remission. 166 treatment-seeking adults with OCD, recruited as part of the Brown Longitudinal Obsessive-Compulsive Study, completed the NEO Five-Factor Inventory 3 (NEO-FFI) and were in episode for OCD at time of NEO-FFI completion. Participants were followed for up to 3 years. Results suggest individuals with OCD had a 21 % likelihood of reaching remission over the course of 3 years. Greater OCD symptom severity and longer illness duration were associated with a decreased likelihood of remission. Among the five factors of personality, only low extraversion was associated with a decreased rate of remission. Neuroticism, openness, agreeableness, and conscientiousness were not associated with remission. As this was an observational study, treatment was not controlled precluding examination of treatment on course. Further, data collected on age of onset and symptom severity during follow up were retrospective and therefore are also subject to recall bias. Our findings provide preliminary support that personality traits are potential factors impacting course and symptom presentation. Future research is necessary to determine the mechanisms in which personality traits may influence the presentation and course of OCD.


A 4-channel telemetry system for hospital patient monitoring.
A 4-channel radio telemetry system has been designed and implemented for use in the operating room on patients undergoing surgery. 30 such systems are in daily use throughout the Duke University Medical Cener. Relative simplicity, long battery life and a DC-response capability are the important features of these systems. The four physiological channels used are; electroencephalogram, electrocardiogram, peripheral pulse, and arterial blood pressure.


Mathematical modelling of miRNA mediated BCR.ABL protein regulation in chronic myeloid leukaemia vis-a-vis therapeutic strategies.
Chronic myeloid leukaemia (CML) is a clonal myeloproliferative disease resulting from an aberrant BCR.ABL gene and protein. To predict BCR.ABL protein abundance and phosphorylation in individual cells in a population of CML cells, we modelled BCR.ABL protein regulation through associated miRNAs using a systems approach. The model rationalizes the level of BCR.ABL protein heterogeneity in CML cells in correlation with the heterogeneous BCR.ABL mRNA levels. We also measured BCR.ABL mRNA and BCR.ABLp phosphorylation in individual cells. The experimental data were consistent with the modelling results, thereby partly validating the model. Provided it is tested further, the model may be used to support effective therapeutic strategies including the combined application of a tyrosine kinase inhibitor and miRNAs targeting BCR.ABL. It appears able to predict different effects of the two types of drug on cells with different expression levels and consequently different effects on the generation of resistance.


Splanchnic perfusion evaluation during hemorrhage and resuscitation with gastric near-infrared spectroscopy.
The purpose of this study was to use a prototype side-illuminating near-infrared spectroscopy (NIRS) nasogastric probe to continuously measure changes in gastric tissue oxygen saturation (Sto2) in a pig hemorrhage model. Swine (n = 12; 6 per group) underwent laparotomy and placement of a gastric NIRS probe, jejunal tonometer, superior mesenteric artery (SMA) flow probe, and a portal vein catheter. Animals underwent hemorrhage (28 mL/kg) t = 0 to 20 minutes (where t = time). Pigs in group I were resuscitated (t = 20-40 minutes) with lactated Ringer's solution (84 mL/kg), whereas group II had no resuscitation. A significant decrease in mean arterial pressure and SMA flow was observed after hemorrhage. SMA flow significantly correlated in group I with both NIRS Sto2 (r = 0.58, p = 0.0001) and regional CO2 (r = -0.54, p = 0.0001). In group II, superior mesenteric flow correlated with NIRS Sto2 (r = 0.30, p = 0.03), but not regional CO2 (r = -0.23, p = 0.09). Direct measurement of tissue oxygen saturation with a prototype side-illuminating near-infrared spectroscopy gastric probe appeared to rapidly reflect changes in splanchnic perfusion.


Understanding Rare Disease Experiences Through the Concept of Morally Problematic Situations.
Rare diseases, defined as having a prevalence inferior to 1/2000, are poorly understood scientifically and medically. Appropriate diagnoses and treatments are scarce, adding to the burden of living with chronic medical conditions. The moral significance of rare disease experiences is often overlooked in qualitative studies conducted with adults living with rare diseases. The concept of morally problematic situations arising from pragmatist ethics shows promise in understanding these experiences. The objectives of this study were to (1) acquire an in-depth understanding of morally problematic situations experienced by adults living with rare diseases in the province of Québec and (2) to develop an integrative model of the concept of morally problematic situations. To this end, an online survey targeting this population was developed through a participatory action research project. Respondents provided 90 long testimonies on the most important morally problematic situations they faced, often in healthcare settings. An integrative model was developed based on various qualitative analyses of these testimonies and relevant literature. The integrative model showcases that morally problematic situations have causes (i.e., contextual and relational factors, personal factors, jeopardized valuations), have affective repercussions (i.e., emotions and feelings, internal tensions), prompt action (i.e., through empowerment strategies leading to the evolution of situations), and elicit outcomes (i.e., factual consequences, residual emotions and feelings, positive or negative resolutions). In sum, this study advances understanding of the moral experiences of adults living with rare diseases while proposing a comprehensive conceptual tool to guide future empirical bioethics research on moral experiences.


Local determinants of road traffic noise levels versus determinants of air pollution levels in a Mediterranean city.
Both traffic-related noise and air pollution have been associated with cardiovascular disease (CVD). Spatial correlations between these environmental stressors may entail mutual confounding in epidemiological studies investigating their long-term effects. Few studies have investigated their correlation - none in Spain - and results differ among cities. We assessed the contribution of urban land-use and traffic variables to the noise-air pollution correlation in Girona town, where an investigation of the chronic effects of air pollution and noise on CVD takes place (REGICOR-AIR). Outdoor annual mean concentrations of nitrogen dioxide (NO(2)) derived from monthly passive sampler measurements were obtained at 83 residential locations. Long-term average traffic-related noise levels from a validated model were assigned to each residence. Linear regression models were fitted both for NO(2) and noise. The correlation between NO(2) and noise (L(24h)) was 0.62. However, the correlation differed across the urban space, with lower correlations at sites with higher traffic density and in the modern downtown. Traffic density, distance from the location to the sidewalk and building density nearby explained 35.6% and 73.2% of the variability of NO(2) and noise levels, respectively. The correlation between the residuals of the two models suggested the presence of other unmeasured common variables. The substantial correlation between traffic-related noise and NO(2), endorsed by common determinants, and the dependence of this correlation on complex local characteristics call for careful evaluations of both factors to ultimately assess their cardiovascular effects.


Pruritus in primary myelofibrosis: clinical and laboratory correlates.
Recent clinical trials with JAK or mammalian target of rapamycin (mTOR) inhibitors in primary myelofibrosis (PMF) have identified pruritus as one of the most treatment-responsive disease traits. However, little is known about the prevalence of pruritus in PMF or its clinical and laboratory correlates. Among 566 consecutive patients with PMF seen at our institution, the presence or absence of pruritus was documented in 90 (16%) and 146 (26%) patients, respectively. Patients with pruritus were less likely to express MPLW515 (0% vs. 10%; P = 0.02) or leukopenia (8% vs. 24%; P = 0.002). The latter association was more pronounced in the absence of JAK2 or MPL mutations. Pruritus also clustered with marked leukocytosis (23% vs. 11%; P = 0.01) and JAK2V617F (71% vs. 59%; P = 0.08). Pruritus did not correlate with karyotype (P = 0.33), risk category per the Dynamic International Prognostic Scoring System (DIPSS)-plus (P = 0.37), DIPSS-plus-adjusted survival (P = 0.41), or leukemic transformation (P = 0.13). Plasma levels of 20 cytokines, which are known to be abnormally expressed in PMF, including IL-1b, IL-2R, IL-6, IL-8, and VEGF, were measured in 63 informative cases and showed no correlations with history of pruritus. We conclude that pruritus is relatively frequent in PMF and is prognostically irrelevant. The pathogenesis of PMF-associated pruritus is not necessarily linked to proinflammatory cytokines but may instead involve molecules that are either granulocyte-derived or influence granulopoiesis. The apparently differential effect of MPL vs. JAK2 mutations on pruritus requires further investigation.


Examination of Reticulocytosis among Chronically Transfused Children with Sickle Cell Anemia.
Sickle cell anemia (SCA) is an inherited hemolytic anemia with compensatory reticulocytosis. Recent studies have shown that increased levels of reticulocytosis during infancy are associated with increased hospitalizations for SCA sequelae as well as cerebrovascular pathologies. In this study, absolute reticulocyte counts (ARC) measured prior to transfusion were analysed among a cohort of 29 pediatric SCA patients receiving chronic transfusion therapy (CTT) for primary and secondary stroke prevention. A cross-sectional flow cytometric analysis of the reticulocyte phenotype was also performed. Mean duration of CTT was 3.1 ± 2.6 years. Fifteen subjects with magnetic resonance angiography (MRA) -vasculopathy had significantly higher mean ARC prior to initiating CTT compared to 14 subjects without MRA-vasculopathy (427.6 ± 109.0 K/μl vs. 324.8 ± 109.2 K/μl, p<0.05). No significant differences in hemoglobin or percentage sickle hemoglobin (HbS) were noted between the two groups at baseline. Reticulocyte phenotyping further demonstrated that the percentages of circulating immature [CD36(+), CD71(+)] reticulocytes positively correlated with ARC in both groups. During the first year of CTT, neither group had significant reductions in ARC. Among this group of children with SCA, cerebrovasculopathy on MRA at initiation of CTT was associated with increased reticulocytosis, which was not reduced after 12 months of transfusions.


Interventions to increase influenza vaccination rates of those 60 years and older in the community.
The effectiveness of interventions to increase influenza vaccination uptake in people aged 60 years and older varies by country and participant characteristics. This review updates versions published in 2010 and 2014. To assess access, provider, system, and societal interventions to increase the uptake of influenza vaccination in people aged 60 years and older in the community. We searched CENTRAL, which includes the Cochrane Acute Respiratory Infections Group's Specialised Register, MEDLINE, Embase, CINAHL, and ERIC for this update, as well as WHO ICTRP and ClinicalTrials.gov for ongoing studies to 7 December 2017. We also searched the reference lists of included studies. Randomised controlled trials (RCTs) and cluster-randomised trials of interventions to increase influenza vaccination in people aged 60 years or older in the community. We used standard methodological procedures as specified by Cochrane. We included three new RCTs for this update (total 61 RCTs; 1,055,337 participants). Trials involved people aged 60 years and older living in the community in high-income countries. Heterogeneity limited some meta-analyses. We assessed studies as at low risk of bias for randomisation (38%), allocation concealment (11%), blinding (44%), and selective reporting (100%). Half (51%) had missing data. We assessed the evidence as low-quality. We identified three levels of intervention intensity: low (e.g. postcards), medium (e.g. personalised phone calls), and high (e.g. home visits, facilitators).Increasing community demand (12 strategies, 41 trials, 53 study arms, 767,460 participants)One successful intervention that could be meta-analysed was client reminders or recalls by letter plus leaflet or postcard compared to reminder (odds ratio (OR) 1.11, 95% confidence interval (CI) 1.07 to 1.15; 3 studies; 64,200 participants). Successful interventions tested by single studies were patient outreach by retired teachers (OR 3.33, 95% CI 1.79 to 6.22); invitations by clinic receptionists (OR 2.72, 95% CI 1.55 to 4.76); nurses or pharmacists educating and nurses vaccinating patients (OR 152.95, 95% CI 9.39 to 2490.67); medical students counselling patients (OR 1.62, 95% CI 1.11 to 2.35); and multiple recall questionnaires (OR 1.13, 95% CI 1.03 to 1.24).Some interventions could not be meta-analysed due to significant heterogeneity: 17 studies tested simple reminders (11 with 95% CI entirely above unity); 16 tested personalised reminders (12 with 95% CI entirely above unity); two investigated customised compared to form letters (both 95% CI above unity); and four studies examined the impact of health risk appraisals (all had 95% CI above unity). One study of a lottery for free groceries was not effective.Enhancing vaccination access (6 strategies, 8 trials, 10 arms, 9353 participants)We meta-analysed results from two studies of home visits (OR 1.30, 95% CI 1.05 to 1.61) and two studies that tested free vaccine compared to patient payment for vaccine (OR 2.36, 95% CI 1.98 to 2.82). We were unable to conduct meta-analyses of two studies of home visits by nurses plus a physician care plan (both with 95% CI above unity) and two studies of free vaccine compared to no intervention (both with 95% CI above unity). One study of group visits (OR 27.2, 95% CI 1.60 to 463.3) was effective, and one study of home visits compared to safety interventions was not.Provider- or system-based interventions (11 strategies, 15 trials, 17 arms, 278,524 participants)One successful intervention that could be meta-analysed focused on payments to physicians (OR 2.22, 95% CI 1.77 to 2.77). Successful interventions tested by individual studies were: reminding physicians to vaccinate all patients (OR 2.47, 95% CI 1.53 to 3.99); posters in clinics presenting vaccination rates and encouraging competition between doctors (OR 2.03, 95% CI 1.86 to 2.22); and chart reviews and benchmarking to the rates achieved by the top 10% of physicians (OR 3.43, 95% CI 2.37 to 4.97).We were unable to meta-analyse four studies that looked at physician reminders (three studies with 95% CI above unity) and three studies of facilitator encouragement of vaccination (two studies with 95% CI above unity). Interventions that were not effective were: comparing letters on discharge from hospital to letters to general practitioners; posters plus postcards versus posters alone; educational reminders, academic detailing, and peer comparisons compared to mailed educational materials; educational outreach plus feedback to teams versus written feedback; and an intervention to increase staff vaccination rates.Interventions at the societal levelNo studies reported on societal-level interventions.Study funding sourcesStudies were funded by government health organisations (n = 33), foundations (n = 9), organisations that provided healthcare services in the studies (n = 3), and a pharmaceutical company offering free vaccines (n = 1). Fifteen studies did not report study funding sources. We identified interventions that demonstrated significant positive effects of low (postcards), medium (personalised phone calls), and high (home visits, facilitators) intensity that increase community demand for vaccination, enhance access, and improve provider/system response. The overall GRADE assessment of the evidence was moderate quality. Conclusions are unchanged from the 2014 review.


Effectiveness of Global, Low-Degree Polynomial Transformations for GCxGC Data Alignment.
As columns age and differ between systems, retention times for comprehensive two-dimensional gas chromatography (GCxGC) may vary between runs. To properly analyze GCxGC chromatograms, it often is desirable to align the retention times of chromatographic features, such as analyte peaks, between chromatograms. Previous work by the authors has shown that global, low-degree polynomial transformation functions, namely affine, second-degree polynomial, and third-degree polynomial, are effective for aligning pairs of two-dimensional chromatograms acquired with dual second columns and detectors (GC×2GC). This work assesses the experimental performance of these global methods on more general GCxGC chromatogram pairs and compares their performance to that of a recent, robust, local alignment algorithm for GCxGC data [ Gros Anal. Chem. 2012 , 84 , 9033 ]. Measuring performance with the root-mean-square (RMS) residual differences in retention times for matched peaks suggests that global, low-degree polynomial transformations outperform the local algorithm given a sufficiently large set of alignment points, and are able to improve misalignment by over 95% based on a lower-bound benchmark of inherent variability. However, with small sets of alignment points, the local method demonstrated lower error rates (although with greater computational overhead). For GCxGC chromatogram pairs with only slight initial misalignment, none of the global or local methods performed well. In some cases with initial misalignment near the inherent variability of the system, these methods worsened alignment, suggesting that it may be better not to perform alignment in such cases.


Insight into the Mechanism of Reversible Ring-Opening of 1,3-Benzoxazine with Thiols.
The reversible ring-opening addition and fragmentation reaction of p-cresol-based N-phenylbenzoxazine with aliphatic and aromatic thiols was investigated in solvent-mediated and solvent-free reactions. Independently of the used thiol, N-phenylbenzoxazine and the thiols reacted to equilibrium with comparable amounts of reactants and products in aprotic solvent, whereas in protic solvent almost full conversions were reached. In contrast, thiol reactivity was a crucial factor in solvent-free reactions yielding fast and complete conversions for a more acidic thiol and balanced equilibrium concentrations in case of thiols with high pKa values. The strong influence of thiols with low pKa values emphasizes the relevance of the protonation step in the ring-opening reactions of 1,3-benzoxazines with thiols in absence of solvents where acidity predominates nucleophilicity. The reverse reactions, namely adduct dissociation and benzoxazine recovery, were successfully conducted at elevated temperatures and reduced pressure facilitated by the removal of the formed thiols yielding up to 95% recovered 1,3-benzoxazine. These results provide deeper understanding of the reversible ring-opening reaction mechanism of 1,3-benzoxazine with thiols.


Molecular cloning of the hepcidin gene from crescent sweetlips (Plectorhinchus cinctus) and characterization of its encoded antimicrobial peptide.
Hepcidin has been identified as an important antimicrobial peptide exerting important innate immunomodulatory activities in many fish species. In the present study, reverse transcription PCR coupled with the rapid amplification of cDNA ends was used to obtain the full-length cDNA of the crescent sweetlips hepcidin gene, which is 829 bp in length and includes an 273 bp ORF encoding a peptide with 90 amino acid residues. Sequence alignment showed a typical RXKR motif and eight conserved cysteine residues in the deduced amino acid sequences. Four disulfide bonds were predicted to form between these eight cysteines, which may stabilize the hairpin structure in hepcidin molecule. Furthermore, phylogenetic analysis showed that the deduced amino acid sequences of crescent sweetlips hepcidin had high sequence homology to hepcidins from fish species of Eupercaria. In addition, the crescent sweetlips hepcidin peptide demonstrated a strong antimicrobial activity in vitro against several types of pathogenic bacteria in fish. In conclusion, the obtained results suggested that crescent sweetlips hepcidin possessed the typical structure similar to other fish hepcidins and had strong antibacterial activity, which showed great potential in the prevention of fish diseases in aquaculture.


Copper arylnitrene intermediates: formation, structure and reactivity.
The mechanism of oxidation of arylamines by copper enzymes is not clarified yet. Here, we explored a reaction between a possible high-valent copper(ii)-oxyl intermediate and arylamine. We have employed a TPA ligand (TPA = tris(2-pyridylmethyl)amine) with the NH2 group in position 2 of one of the pyridine rings (TPANH2). This model system allows generation of [(TPANH2)Cu(O)]+ in the gas phase, which immediately undergoes a reaction between the arylamino group and the copper oxyl moiety. The reaction leads to elimination of H2O and formation of a copper-nitrene complex. The structure of the resulting copper-nitrene complex was confirmed by infrared spectroscopy in the gas phase. We show that the copper-nitrene complex reacts by hydrogen atom transfer with 1,4-cyclohexadiene and by an order of magnitude faster by a double hydrogen atom transfer with ethanethiol and methanol. DFT calculations explain the formation of the copper nitrene as well as its reactivity in agreement with the experimental findings.


Visual marking in moving displays: feature-based inhibition is not necessary.
Visual marking is a mechanism by which new visual stimuli can gain a selection advantage by the top-down attentional inhibition of stimuli already in the field. Previous work (Olivers, Watson, & Humphreys, 1999) has shown that, for moving stimuli, there must be a unique feature difference between the old items and the new items for marking to occur. The present study shows that this constraint is not necessary if the local spatial relationships between the old moving items remain constant. It is proposed that, with a fixed configuration, the old moving items can be grouped to form a single object. An inhibitory template set up to represent the object then coordinates the application of inhibition to the individual stimuli. Implications for the theory and ecological flexibility of visual marking are discussed.


Periostin Limits Tumor Response to VEGFA Inhibition.
Resistance to antiangiogenic drugs limits their applicability in cancer therapy. Here, we show that revascularization and progression of pancreatic neuroendocrine tumors (PNETs) under extended vascular-endothelial growth factor A (VEGFA) blockade are dependent on periostin (POSTN), a matricellular protein expressed by stromal cells. Genetic deletion of Postn in RIP1-Tag2 mice blunted tumor rebounds of M2-like macrophages and αSMA+ stromal cells in response to prolonged VEGFA inhibition and suppressed PNET revascularization and progression on therapy. POSTN deficiency also impeded the upregulation of basic fibroblast growth factor (FGF2), an adaptive mechanism previously implicated in PNET evasion from antiangiogenic therapy. Higher POSTN expression correlated with markers of M2-like macrophages in human PNETs, and depleting macrophages with a colony-stimulating factor 1 receptor (CSF1R) antibody inhibited PNET revascularization and progression under VEGFA blockade despite continued POSTN production. These findings suggest a role for POSTN in orchestrating resistance to anti-VEGFA therapy in PNETs.


Autosomal recessive spastic ataxia of Charlevoix Saguenay (ARSACS): expanding the genetic, clinical and imaging spectrum.
Mutations in SACS, leading to autosomal-recessive spastic ataxia of Charlevoix-Saguenay (ARSACS), have been identified as a frequent cause of recessive early-onset ataxia around the world. Here we aimed to enlarge the spectrum of SACS mutations outside Quebec, to establish the pathogenicity of novel variants, and to expand the clinical and imaging phenotype. Sequencing of SACS in 22 patients with unexplained early-onset ataxia, assessment of novel SACS variants in 3.500 European control chromosomes and extensive phenotypic investigations of all SACS carriers. We identified 11 index patients harbouring 17 novel SACS variants. 9/11 patients harboured two variants of at least probable pathogenicity which were not observed in controls and, in case of missense mutations, were located in highly conserved domains. These 9 patients accounted for at least 11% (9/83) in our series of unexplained early onset ataxia subjects. While most patients (7/9) showed the classical ARSACS triad, the presenting phenotype reached from pure neuropathy (leading to the initial diagnosis of Charcot-Marie-Tooth disease) in one subject to the absence of any signs of neuropathy in another. In contrast to its name "spastic ataxia", neither spasticity (absent in 2/9=22%) nor extensor plantar response (absent in 3/9=33%) nor cerebellar ataxia (absent in 1/9=11%) were obligate features. Autonomic features included urine urge incontinence and erectile dysfunction. Apart from the well-established MRI finding of pontine hypointensities, all patients (100%) showed hyperintensities of the lateral pons merging into the (thickened) middle cerebellar peduncles. In addition, 63% exhibited bilateral parietal cerebral atrophy, and 63% a short circumscribed thinning of the posterior midbody of the corpus callosum. In 2 further patients with differences in important clinical features, VUS class 3 variants (c.1373C>T [p.Thr458Ile] and c.2983 G>T [p.Val995Phe]) were identified. These variants were, however, also observed in controls, thus questioning their pathogenic relevance. We here demonstrate that each feature of the classical ARSACS triad (cerebellar ataxia, spasticity and peripheral neuropathy) might be missing in ARSACS. Nevertheless, characteristic MRI features - which also extend to supratentorial regions and involve the cerebral cortex - will help to establish the diagnosis in most cases.


Developmental regulation of pyrroline-5-carboxylate reductase gene expression in Arabidopsis.
At-P5R, a gene encoding the last enzyme of the proline (Pro) biosynthetic pathway in Arabidopsis thaliana, is developmentally regulated. To characterize the cis elements responsible for this developmental regulation, a series of 5' deletions of the At-P5R promoter were transcriptionally fused to a beta-glucuronidase (GUS)-coding region and transformed into Arabidopsis. The complete promoter of At-P5R directs strong GUS activity in root tips, the shoot meristem, guard cells, hydathodes, pollen grains, ovules, and developing seeds, all of which contain rapidly dividing cells and/or are undergoing changes in osmotic potential. This expression pattern is consistent with the function of Pro as an energy, nitrogen, and carbon source and as an osmoticum in response to dehydration. Promoters longer than 212 base pairs (bp) showed the same expression pattern, whereas those shorter than 143 bp did not direct any detectable GUS activity in any organs. This suggests that a 69-bp promoter region located between -212 and -143 bp is necessary to establish the tissue-specific expression of At-P5R during development. The Pro content measured in different organs suggests that, in addition to transcriptional control of the biosynthetic pathway, the transport of Pro may play a role in its distribution within Arabidopsis. Several aspects of the relationship between Pro metabolism and plant physiology are discussed.


Quantifying the interplay effect in prostate IMRT delivery using a convolution-based method.
The authors present a segment-based convolution method to account for the interplay effect between intrafraction organ motion and the multileaf collimator position for each particular segment in intensity modulated radiation therapy (IMRT) delivered in a step-and-shoot manner. In this method, the static dose distribution attributed to each segment is convolved with the probability density function (PDF) of motion during delivery of the segment, whereas in the conventional convolution method ("average-based convolution"), the static dose distribution is convolved with the PDF averaged over an entire fraction, an entire treatment course, or even an entire patient population. In the case of IMRT delivered in a step-and-shoot manner, the average-based convolution method assumes that in each segment the target volume experiences the same motion pattern (PDF) as that of population. In the segment-based convolution method, the dose during each segment is calculated by convolving the static dose with the motion PDF specific to that segment, allowing both intrafraction motion and the interplay effect to be accounted for in the dose calculation. Intrafraction prostate motion data from a population of 35 patients tracked using the Calypso system (Calypso Medical Technologies, Inc., Seattle, WA) was used to generate motion PDFs. These were then convolved with dose distributions from clinical prostate IMRT plans. For a single segment with a small number of monitor units, the interplay effect introduced errors of up to 25.9% in the mean CTV dose compared against the planned dose evaluated by using the PDF of the entire fraction. In contrast, the interplay effect reduced the minimum CTV dose by 4.4%, and the CTV generalized equivalent uniform dose by 1.3%, in single fraction plans. For entire treatment courses delivered in either a hypofractionated (five fractions) or conventional (> 30 fractions) regimen, the discrepancy in total dose due to interplay effect was negligible.


Effects of feed energy density, daily milking frequency, and a single injection of cabergoline on behavior and welfare in dairy cows at dry-off.
Drying off dairy cows may challenge animal welfare due to high milk yields. A total of 111 loose-housed Holstein cows yielding >15 kg/d of milk were included in a 2 × 2 × 2 factorial design during dry-off to investigate the effects of reduced feeding level (normal vs. reduced energy density), reduced milking frequency (twice vs. once daily), and administration of a dopamine agonist (saline i.m. injection vs. cabergoline i.m. injection) on behavior in the home pen. During the 7 d before dry-off, cows were fed and milked according to 1 of the 4 feeding level and milking frequency combinations. Within 3 h after the last milking, cows were injected i.m. with 5 mL of either saline or a dopamine agonist (5.6 mg of cabergoline; Velactis, Ceva Santé Animale, Libourne, France; labeled for use only with abrupt dry-off, i.e., no preceding reduction in feeding level or milking frequency before last milking). Cows' behavior during d -1, 0, and +1 relative to the last milking was recorded via video and leg-attached sensors. Cows on the reduced energy density diet spent more time feeding and showed more attempts to feed from other cows' bins on d -1. Throughout the period of observations, cows on the reduced diet spent a lower percentage of lying time with their head raised, a higher percentage of lying time with their legs bent, and less time standing in a vigilant posture than did cows on the normal lactation diet. Reducing the daily milking frequency from 2 to 1 did not result in any clear behavioral signs of discomfort. On d 0, cows injected with cabergoline lay down longer but had their head raised for a shorter percentage of time while lying, compared with cows injected with saline. Cows injected with cabergoline also spent less time feeding than cows injected with saline on d 0, and reduced the time spent drinking from d -1 to d 0. Finally, fewer cabergoline-injected cows used the brush for self-grooming, and, among cows that did use the brush, the cows injected with cabergoline reduced the time spent using the brush from d -1 to d 0. In conclusion, cows injected with cabergoline showed several behavioral changes compared with control cows injected with saline. The behavioral changes shown by cows injected with cabergoline may be indicative of malaise during the first 24 h after injection, raising concern for animal welfare. No behavioral evidence for reduced udder pain in cows injected with cabergoline compared with control cows injected with saline was found. Drying off by reducing the energy density of the diet caused behavioral changes indicative of hunger before dry-off, whereas reducing the milking frequency had no clear effects on behavior.


A serotonin-induced N-glycan switch regulates platelet aggregation.
Serotonin (5-HT) is a multifunctional signaling molecule that plays different roles in a concentration-dependent manner. We demonstrated that elevated levels of plasma 5-HT accelerate platelet aggregation resulting in a hypercoagulable state in which the platelet surface becomes occupied by several glycoproteins. Here we study the novel hypothesis that an elevated level of plasma 5-HT results in modification of the content of N-glycans on the platelet surface and this abnormality is associated with platelet aggregation. Mass spectrometry of total surface glycoproteins on platelets isolated from wild-type mice infused for 24 hours with saline or 5-HT revealed that the content of glycoproteins on platelets from 5-HT-infused mice switched from predominantly N-acetyl-neuraminic acid (Neu5Ac) to N-glycolyl-neuraminic acid (Neu5Gc). Cytidine monophosphate-N-acetylneuraminate hydroxylase (CMAH) synthesizes Neu5Gc from Neu5Ac. Up-regulation of Neu5Gc content on the platelet surface resulted from an increase in the catalytic function, not expression, of CMAH in platelets of 5-HT-infused mice. The highest level of Neu5Gc was observed in platelets of 5-HT-infused, 5-HT transporter-knock out mice, suggesting that the surface delineated 5-HT receptor on platelets may promote CMAH catalytic activity. These new findings link elevated levels of plasma 5-HT to altered platelet N-glycan content, a previously unrecognized abnormality that may favor platelet aggregation.


Very low dose naltrexone addition in opioid detoxification: a randomized, controlled trial.
Although current treatments for opioid detoxification are not always effective, medical detoxification remains a required step before long-term interventions. The use of opioid antagonist medications to improve detoxification has produced inconsistent results. Very low dose naltrexone (VLNTX) was recently found to reduce opioid tolerance and dependence in animal and clinical studies. We decided to evaluate safety and efficacy of VLNTX adjunct to methadone in reducing withdrawal during detoxification. In a multi-center, double-blind, randomized study at community treatment programs, where most detoxifications are performed, 174 opioid-dependent subjects received NTX 0.125 mg, 0.250 mg or placebo daily for 6 days, together with methadone in tapering doses. VLNTX-treated individuals reported attenuated withdrawal symptoms [F = 7.24 (2,170); P = 0.001] and reduced craving [F = 3.73 (2,107); P = 0.03]. Treatment effects were more pronounced at discharge and were not accompanied by a significantly higher retention rate. There were no group differences in use of adjuvant medications and no treatment-related adverse events. Further studies should explore the use of VLNTX, combined with full and partial opioid agonist medications, in detoxification and long-term treatment of opioid dependence.


Isolation of a large number of novel mammalian genes by a differential cDNA library screening strategy.
As part of the ongoing human and mouse genome projects, the aim of this study was to isolate novel, previously uncharacterized, genes from mouse testis. Two approaches were compared for their effectiveness in isolating novel genes: random, vs differential, complementary DNA (cDNA) cloning methods. In the differential approach, only the cDNA clones containing rare sequences (as determined by preliminary clone hybridization) are further analyzed; in the random approach, cDNA clones are isolated at random from the cDNA library. More than two hundred cDNA clones altogether were analyzed, using a PCR-mediated amplification and sequencing strategy. A comparison of these sequences to nucleic acid and protein sequence databases, revealed that 84% of the isolated rare cDNA clones represented new, previously uncharacterized mouse genes. In contrast, less than 63% of the cDNA clones isolated at random from cDNA libraries, contained novel genes. Thus, the probability of isolating new, previously uncharacterized, mammalian genes from cDNA libraries can be markedly improved by focusing efforts on clones containing rare sequences.


What data determine whether a plant taxon is distinct enough to merit legal protection? A case study of Sedum integrifolium (Crassulaceae).
Measures of molecular and morphological genetic variation are often used to set conservation priorities and design management strategies for plant taxa. Evaluated together they can give insights into a taxon's evolutionary status that neither data type alone can achieve. We investigated the distinctness and variability of Sedum integrifolium ssp. leedyi, a federally and state-listed taxon, from its conspecific relatives using 33 random amplified polymorphic DNA (RAPD) markers (253 plants) and 37 morphological characters from 1308 common-garden-grown plants. We included S. integrifolium ssp. leedyi (four populations), its conspecific relatives (11 populations), and S. rosea and S. rhodanthum populations in our study. The morphological and molecular data correspond in showing that S. integrifolium ssp. leedyi populations are highly distinct. However, the data sets differ in their estimates of the relatedness of some S. integrifolium ssp. leedyi populations and in the percentage variation detected due to differences among them (25 and 9-13% for the molecular and morphological data, respectively) suggesting little gene flow among populations and some differentiation, possibly from selective pressures. Given our data, we recommend that S. integrifolium ssp. leedyi merits protection under the U.S. Endangered Species Act and that its populations be managed as distinct units.


[Surgery of craniofacial deformities].
Most craniofacial abnormalities are craniosynostoses due to premature fusion of one or more craniofacial sutures. The typical cranial deformities result from growth inhibition perpendicular to the affected sutures. Functional impairment is caused either directly by the pathological growth pattern or indirectly by the increased intracranial pressure. Craniosynostosis is frequently accompanied by dysmorphia or dystopia of the orbits, resulting in possible functional problems. The indications for operation are to increase the intracranial volume and to correct aesthetic impairments. The planning of the surgical procedures has to consider the orbital problems. This review describes the different disease patterns of premature craniosynostosis and specific surgical approaches for their correction.


A novel pathway for amyloids self-assembly in aggregates at nanomolar concentration mediated by the interaction with surfaces.
A limitation of the amyloid hypothesis in explaining the development of neurodegenerative diseases is that the level of amyloidogenic polypeptide in vivo is below the critical concentration required to form the aggregates observed in post-mortem brains. We discovered a novel, on-surface aggregation pathway of amyloidogenic polypeptide that eliminates this long-standing controversy. We applied atomic force microscope (AFM) to demonstrate directly that on-surface aggregation takes place at a concentration at which no aggregation in solution is observed. The experiments were performed with the full-size Aβ protein (Aβ42), a decapeptide Aβ(14-23) and α-synuclein; all three systems demonstrate a dramatic preference of the on-surface aggregation pathway compared to the aggregation in the bulk solution. Time-lapse AFM imaging, in solution, show that over time, oligomers increase in size and number and release in solution, suggesting that assembled aggregates can serve as nuclei for aggregation in bulk solution. Computational modeling performed with the all-atom MD simulations for Aβ(14-23) peptide shows that surface interactions induce conformational transitions of the monomer, which facilitate interactions with another monomer that undergoes conformational changes stabilizing the dimer assembly. Our findings suggest that interactions of amyloidogenic polypeptides with cellular surfaces play a major role in determining disease onset.


The maize glossy13 gene, cloned via BSR-Seq and Seq-walking encodes a putative ABC transporter required for the normal accumulation of epicuticular waxes.
Aerial plant surfaces are covered by epicuticular waxes that among other purposes serve to control water loss. Maize glossy mutants originally identified by their "glossy" phenotypes exhibit alterations in the accumulation of epicuticular waxes. By combining data from a BSR-Seq experiment and the newly developed Seq-Walking technology, GRMZM2G118243 was identified as a strong candidate for being the glossy13 gene. The finding that multiple EMS-induced alleles contain premature stop codons in GRMZM2G118243, and the one knockout allele of gl13, validates the hypothesis that gene GRMZM2G118243 is gl13. Consistent with this, GRMZM2G118243 is an ortholog of AtABCG32 (Arabidopsis thaliana), HvABCG31 (barley) and OsABCG31 (rice), which encode ABCG subfamily transporters involved in the trans-membrane transport of various secondary metabolites. We therefore hypothesize that gl13 is involved in the transport of epicuticular waxes onto the surfaces of seedling leaves.


Contrast-Enhanced Computed Tomography Enables Quantitative Evaluation of Tissue Properties at Intrajoint Regions in Cadaveric Knee Cartilage.
Objective The aim of this study was to investigate whether the concentration of the anionic contrast agent ioxaglate, as quantitated by contrast-enhanced computed tomography (CECT) using a clinical cone-beam CT (CBCT) instrument, reflects biochemical, histological, and biomechanical characteristics of articular cartilage imaged in an ex vivo, intact human knee joint. Design An osteoarthritic human cadaveric knee joint (91 years old) was injected with ioxaglate (36 mg I/mL) and imaged using CBCT over 61 hours of ioxaglate diffusion into cartilage. Following imaging, the joint surfaces were excised, rinsed to remove contrast agent, and compressive stiffness (equilibrium and instantaneous compressive moduli) was measured via indentation testing ( n = 17 sites). Each site was sectioned for histology and assessed for glycosaminoglycan content using digital densitometry of Safranin-O stained sections, Fourier transform infrared spectroscopy for collagen content, and morphology using both the Mankin and OARSI semiquantitative scoring systems. Water content was determined using mass change after lyophilization. Results CECT attenuation at all imaging time points, including those <1 hour of ioxaglate exposure, correlated significantly ( P < 0.05) with cartilage water and glycosaminoglycan contents, Mankin score, and both equilibrium and instantaneous compressive moduli. Early time points (<30 minutes) also correlated ( P < 0.05) with collagen content and OARSI score. Differences in cartilage quality between intrajoint regions were distinguishable at diffusion equilibrium and after brief ioxaglate exposure. Conclusions CECT with ioxaglate affords biochemical and biomechanical measurements of cartilage health and performance even after short, clinically relevant exposure times, and may be useful in the clinic as a means for detecting early signs of cartilage pathology.


Rapidly Progressing Neurocognitive Disorder in a Male with FXTAS and Alzheimer's Disease.
Fragile X-associated tremor/ataxia syndrome (FXTAS) is a neurodegenerative disorder that usually begins in the early 60s and affects carriers of premutation expansion (55-200 CGG repeats) of the fragile X mental retardation 1 (FMR1) gene. Additional disorders can co-occur with FXTAS including Alzheimer's disease (AD). Here we discuss a case report of a male with 67 CGG repeats in FMR1 who had mild late-onset FXTAS symptoms followed by neurocognitive disorder symptoms consistent with AD. The patient has developed tremor and ataxia that are the two characteristic symptoms of FXTAS. In addition, he shows rapid cognitive decline, brain atrophy most substantial in the medial temporal lobe, and decreased metabolism in the brain regions that are the characteristic findings of AD. The purpose of this study is to describe a patient profile with both diseases and review the details of an overlap between these two diseases.


IL-1β-mediated signals preferentially drive conversion of regulatory T cells but not conventional T cells into IL-17-producing cells.
Regulatory T cells (Tregs) are committed to suppressive functions. Recently, it was proposed that Tregs could produce IL-17 under proinflammatory, polarizing conditions. We studied the role of Tregs on IL-17 production in the absence of exogenous cytokines and insults. Using in vitro and in vivo approaches, we determined that under neutral conditions, simultaneous activation of Tregs and naive CD4(+) conventional T cells in the presence of APCs resulted in conversion of Tregs into IL-17-producing cells, and endogenous IL-1β was mandatory in this process. Mechanistic analysis revealed that the IL-1R1 was highly expressed on Tregs and that IL-1β induced marked activation of p38 and JNK, which were involved in IL-17 production. These observations could have important implications on therapeutic strategies using Tregs.


Analyzing the Subspace Structure of Related Images: Concurrent Segmentation of Image Sets.
We develop new algorithms to analyze and exploit the joint subspace structure of a set of related images to facilitate the process of concurrent segmentation of a large set of images. Most existing approaches for this problem are either limited to extracting a single similar object across the given image set or do not scale well to a large number of images containing multiple objects varying at different scales. One of the goals of this paper is to show that various desirable properties of such an algorithm (ability to handle multiple images with multiple objects showing arbitary scale variations) can be cast elegantly using simple constructs from linear algebra: this significantly extends the operating range of such methods. While intuitive, this formulation leads to a hard optimization problem where one must perform the image segmentation task together with appropriate constraints which enforce desired algebraic regularity (e.g., common subspace structure). We propose efficient iterative algorithms (with small computational requirements) whose key steps reduce to objective functions solvable by max-flow and/or nearly closed form identities. We study the qualitative, theoretical, and empirical properties of the method, and present results on benchmark datasets.


Oil-free eye drops containing Cyclosporine A/cyclodextrin/PVA supramolecular complex as a treatment modality for dry eye disease.
According to the global mapping of dry eye disease (DED), nearly 5 to 50 % of people suffer from DED, and this number is on the rise. The drug of choice Cyclosporine A (CsA) exhibits poor ocular bioavailability due to high molecular weight and lipophilicity. Moreover, formulations of CsA currently available are in the form of oil-based emulsions that are known to cause ocular irritation and pain. In this study, sulfobutylether-β-cyclodextrin (SBE-β-CD) based binary and ternary supramolecular complexes of CsA were developed as completely oil-free, and particle-free eye drops to treat DED. The physicochemical characterizations were supplemented with relevant in silico studies, to ascertain the findings. Further, the efficacy of the complexes was evaluated in the scopolamine-induced mouse model of DED. The complexation improved the CsA solubility by ~21-fold, with ~4-fold improvement in dissolution and transcorneal permeation. The non-irritancy and non-toxicity were confirmed by hen's egg chorioallantoic membrane assay and cytotoxicity assay using human corneal epithelial cells, respectively. The in vivo treatment with the ternary CD complex demonstrated better management of the dry eye supported by the tear volume assessment, corneal fluorescein staining, and histopathological studies of the cornea, lacrimal gland, and harderian gland. The study demonstrates the potential of the supramolecular complex as an alternative to the oil-based formulation of eye drops for drugs that show low solubility and poor corneal permeation.


Short course therapy with cefuroxime axetil for acute otitis media: results of a randomized multicenter comparison with amoxicillin/clavulanate.
Otitis media is a common infection of childhood. Increasing antibiotic resistance rates among the principal causative pathogens, Streptococcus pneumoniae and Haemophilus influenzae, are associated with failure of first line agents. This open, randomized, multicenter study compared the clinical efficacy of a short 5-day course of cefuroxime axetil (CAE) suspension with that of amoxicillin/clavulanate (A/CA) suspension for 8 or 10 days. Children age 6 to 36 months with acute otitis media with effusion, diagnosed by tympanocentesis and microbiologic culture, were randomized to receive CAE (30 mg/kg/day in two divided doses for 5 days) or A/CA 40 mg/kg/day in three divided doses for 10 days (A/CA-10). In French centers A/CA was given at 80 mg/kg/day in three divided doses for 8 days (A/CA-8). Patients were assessed 1 to 4 days after completing the course (posttreatment) and followed up at 21 to 28 days after completing the course. Of the 716 patients randomized, 252 were treated with CAE, 255 with A/CA-10 and 209 with A/CA-8. In the clinically evaluable population, the proportions of patients with clinical cure at posttreatment were 175 of 203 (86%), 181 of 205 (88%) and 145 of 164 (88%) in the CAE, A/CA-10 and A/CA-8 groups, respectively, demonstrating equivalence among the three treatments. For patients <18 months old, clinical cures were 111 of 134 (83%), 116 of 131 (89%) and 83 of 99 (84%) in the CAE, A/CA-10 and A/CA-8 groups, respectively; equivalence was also demonstrated. At follow-up, 130 of 175 (74%) CAE, 121 of 172 (70%) A/CA-10, and 112 of 142 (79%) A/CA-8 had maintained cure. A total of 837 pretreatment pathogens were isolated from middle ear fluid in 73% (522 of 716) patients, the majority of isolates were S. pneumoniae (30%) and H. influenzae (27%). The most common adverse events were gastrointestinal, the incidence of drug-related diarrhea being higher in the A/CA-10 group (18%) than in either the CAE or A/CA-8 groups (10%). A 5-day course of CAE, given twice daily, was shown to be equivalent to the two regimens of A/CA for treatment of acute otitis media with effusion in children.


Dermatology ECHO: A case presentation demonstrating benefits of specialty telementoring in primary care.
Allergic contact dermatitis (ACD) is a common dermatologic disorder that is estimated to affect 15-20% of the general population. Because of its prevalence, it may be expected that ACD should be easily recognized. However, it can present with many clinical variations that may complicate diagnosis. Although ACD is a treatable condition, patients from rural and underserved areas suffer if timely access to specialty care is limited. Dermatology Extension for Community Healthcare Outcomes (Dermatology ECHO) telemedicine sessions were created to mentor rural primary care providers (PCPs). To illustrate their benefit, we present the case of a 19-year-old female patient who suffered from worsening undiagnosed ACD for over nine months following a laparoscopic appendectomy. During that time, the surgeon and multiple PCPs treated her with antibiotics, antivirals, and Scabicide without improvement in her condition. The de-identified patient case was presented by her PCP during the Dermatology ECHO session. The Dermatology ECHO specialty team mentored and educated the PCP in the diagnosis and treatment of ACD. After making the diagnosis, the patient received new treatment and her condition improved significantly. Dermatology ECHO provides a knowledge-sharing network for participating PCPs that may improve patient outcomes and reduce patient suffering.


Detecting resting-state functional connectivity in the language system using functional near-infrared spectroscopy.
Functional connectivity has become one of the important approaches to understanding the functional organization of the human brain. Recently, functional near-infrared spectroscopy (fNIRS) was demonstrated as a feasible method to study resting-state functional connectivity (RSFC) in the sensory and motor systems. However, whether such fNIRS-based RSFC can be revealed in high-level and complex functional systems remains unknown. In the present study, the feasibility of such an approach is tested on the language system, of which the neural substrates have been well documented in the literature. After determination of a seed channel by a language localizer task, the correlation strength between the low frequency fluctuations of the fNIRS signal at the seed channel and those at all other channels is used to evaluate the language system RSFC. Our results show a significant RSFC between the left inferior frontal cortex and superior temporal cortex, components both associated with dominant language regions. Moreover, the RSFC map demonstrates left lateralization of the language system. In conclusion, the present study successfully utilized fNIRS-based RSFC to study a complex and high-level neural system, and provides further evidence for the validity of the fNIRS-based RSFC approach.


Review on fluoride varnishes currently recommended in dental prophylaxis.
In dentistry, fluoride compounds play a very important role in the development of teeth hard tissue. They have been modifying the development of the carious process for many years in accordance with the principles of minimally invasive therapy. Studies have confirmed their effectiveness in the prevention and treatment of carious lesions and erosion of deciduous and permanent teeth, as well as in the dentin hypersensitivity treatment. Typically, each varnish consists of 3 basic components, i.e., a resin usually in the form of mastic, shellac and/or rosin, an alcohol-based organic solvent (usually ethanol) and active agents. In the first-generation varnishes, the active agent is fluorine compounds, most often in the form of 5% NaF, while in second-generation varnishes, the composition is further enriched with calcium and phosphorus compounds in the form of CPP-ACP/CPP-ACPF, ACP, TCP, fTCP, CSPS, TMP, CXP, or CaGP. This influences the bioavailability of fluoride in the oral environment by increasing both its release from the product and its subsequent accumulation in enamel and plaque, promotes more efficient closure of dentinal tubules, and facilitates pH buffering in the oral cavity.


Bi-level feature selection in high dimensional AFT models with applications to a genomic study.
We propose a new bi-level feature selection method for high dimensional accelerated failure time models by formulating the models to a single index model. The method yields sparse solutions at both the group and individual feature levels along with an expedient algorithm, which is computationally efficient and easily implemented. We analyze a genomic dataset for an illustration, and present a simulation study to show the finite sample performance of the proposed method.


An improved protocol for the production of AFLP markers in complex genomes by means of capillary electrophoresis.
The amplified fragment-length polymorphism (AFLP) technology is a recently introduced method to investigate genomes of different complexity, from microbial to higher organisms. It is applied to purposes as diverse as identification of species, strain and varieties, investigation of genetic diversity within and between populations, simple and complex trait mapping, and construction of linkage and physical maps. This technology has been designed on the use of primers labelled with radioactivity and on AFLP fragment separation on sequencing gel. We show that the original EcoRI/TaqI AFLP protocol does not perform appropriately when transferred to fluorescent labelling and capillary electrophoresis (CE), and propose an improved protocol for the production of high-quality AFLP markers in fish, rodents and artiodactyles by means of the Beckman-Coulter CEQ2000 automatic DNA sequencer. In addition, we describe the procedure routinely used in our laboratory to obtain binary matrices from AFLP profiles with the aid of Genographer free-share software (vers. 1.6.0, J.J. Benham, Montana State University), able to elaborate original fragment data and convert them to standard graphical formats for phylogenetic analyses. Comparison with radioactive AFLPs in goats confirmed the reliability of the protocol developed for CE. In fact, 107 fragments generated by two primer combinations and identified by both techniques were attributed the same scoring. Compared with traditional methods, the use of capillary systems and automated analysis increases data throughput and scoring reliability, decreasing the overall experimental error.


Bladder pain syndrome/interstitial cystitis ESSIC type 3C: high expression of inducible nitric oxide synthase in inflammatory cells.
Bladder pain syndrome/interstitial cystitis (BPS/IC) includes a heterogeneous collection of underlying pathological conditions. Compared to the classic IC with a Hunner lesion, now denominated ESSIC type 3C, the non-Hunner type of BPS/IC appears different in a number of respects. In a previous study, measuring luminal nitric oxide (NO) in the bladder of patients with BPS/IC, it was reported that all patients with ESSIC type 3C had high levels of NO. The aim of the present study was to investigate the source of inducible nitric oxide synthase (iNOS) and thereby the cellular origin of NO production via iNOS. Immunohistochemistry, with two different anti-iNOS antibodies, was used to study 10 patients with BPS/IC ESSIC type 3C who expressed high levels of intraluminal NO. These results were compared with four patients with non-Hunner BPS/IC. To substantiate further the involvement of iNOS in this condition, the protein expression of nitrotyrosine, a marker for iNOS activation, was also assessed. On routine histopathology, the tissues of type 3C patients exhibited inflammatory infiltrates of varying intensity. Strong immunoreactivity for both iNOS and nitrotyrosine was noted within the urothelium but also within the inflammatory infiltrates in the lamina propria of these subjects. The findings of a clearly detectable protein expression of iNOS in both the urothelium and the inflammatory infiltrates in bladder biopsies from patients with BPS/IC ESSIC type 3C suggest that the production of NO, in this entity, may occur in different tissue compartments.


Genetic redundancy in soybean photoresponses associated with duplication of the phytochrome A gene.
Gene and genome duplications underlie the origins of evolutionary novelty in plants. Soybean, Glycine max, is considered to be a paleopolyploid species with a complex genome. We found multiple homologs of the phytochrome A gene (phyA) in the soybean genome and determined the DNA sequences of two paralogs designated GmphyA1 and GmphyA2. Analysis of the GmphyA2 gene from the lines carrying a recessive allele at a photoperiod insensitivity locus, E4, revealed that a Ty1/copia-like retrotransposon was inserted in exon 1 of the gene, which resulted in dysfunction of the gene. Mapping studies suggested that GmphyA2 is encoded by E4. The GmphyA1 gene was mapped to a region of linkage group O, which is homeologous to the region harboring E4 in linkage group I. Plants homozygous for the e4 allele were etiolated under continuous far red light, but the de-etiolation occurred partially, indicating that the mutation alone did not cause a complete loss of phyA function. The genetic redundancy suggests that the presence of duplicated copies of phyA genes accounts for the generation of photoperiod insensitivity, while protecting against the deleterious effects of mutation. Thus, this phenomenon provides a link between gene duplication and establishment of an adaptive response of plants to environments.


The effects of posterior talar glide with dorsiflexion of the ankle on mobility, muscle strength and balance in stroke patients: a randomised controlled trial.
[Purpose] The purpose of this study was to examine the effectiveness of posterior talar glide (PTG) with dorsiflexion of the ankle on stroke patients ankle mobility, muscle strength, and balance ability. [Subjects and Methods] Thirty-four subjects were randomly assigned to either a PTG with dorsiflexion group (PTG; n=17), or a weight-bearing with placebo PTG group (control; n=17). Subjects in the PTG group performed PTG with dorsiflexion, designed to improve ankle mobility, muscle strength and balance ability with proprioceptive control of the ankle, for 10 glides of 5 sets/day, 5 days/week, for 4 weeks. [Results] The experimental group showed significant improvement on the Ankle Dorsiflexion Range of Motion assessment, Ankle Dorsiflexor Manual Muscle Test, Functional Reach Test, Time Up and Go test, and Functional Gait Assessment compared to the control group. However, regarding Ankle Plantarflexion Range of Motion assessment and the Ankle Plantarflexor Manual Muscle Test, no significant differences were found between the two groups. [Conclusion] The results of this study show that PTG with dorsiflexion can improve ankle mobility, muscle strength and balance ability in patients recovering from stroke. This exercise may prove useful in clinical rehabilitation. Further research on the long-term effectiveness of PTG on gait ability is suggested.


Programmed visual field testing.
The use of programmed static spot-checking is remarkably accurate in detecting the presence of visual field defects, even the earliest abnormality, provided the correct stimulus (or stimuli) is chosen to check an appropriate number of spots, and the location of each missed response is retested. Automatic quantitation of visual field defects (so it can later be determined if the defects are worsening) can be accomplished automatically with some of the more expensive new instruments. The less elaborate instruments require repetitive testing or manual interaction, achieving accurate results, but typically with a cumbersome record of the results that is difficult to interpret. An efficient plan for the usual office at present is to use an automated spot-checking instrument for screening and diagnosis, but to depend upon the tangent screen or Goldmann perimeter for quantitation unless the most expensive automated instruments can be afforded.


Ratiometric Optical and Photoacoustic Imaging In Vivo in the Second Near-Infrared Window.
ConspectusOptical imaging and photoacoustic (PA) imaging have become essential tools to investigate physiological or pathological processes at the molecular level in vivo. The detection of variations at the molecular level in vivo is particularly important owing to the rapid progression of diseases. However, most studies have mainly focused on plain qualitative molecular imaging and detection, which is characterized by the absence of a reference signal in one-channel responsive imaging. To overcome the limitation and quantitatively detect molecules in situ, this Account reviews the recent contributions of our group to the quantitative imaging field in the form of ratiometric optical and PA imaging in vivo in the second near-infrared window (NIR-II, 950-1700 nm).In this Account, we present recent advances that our group has made in ratiometric imaging probe design and biomedical applications by constructing probes based on ratiometric optical imaging and ratiometric PA imaging. First, we highlight the design strategies of ratiometric optical probes that were based on organic ratiometric molecular probes, radio-activated organic ratiometric probes, and hybrid organic-inorganic assembled ratiometric probes. Subsequently, the design strategies of the ratiometric NIR-II optical nanoprobes with activated bioluminescence resonance energy transfer (BRET), Förster resonance energy transfer (FRET), and nonradiative energy transfer (NRET) effects provide a reliable tool to achieve the ratiometric detection of endogenous signaling molecules and thereby apply it to the monitoring and evaluation of the efficacy of photodynamic therapy, radiotherapy, and immunotherapy to guide the treatment process. In addition, we systematically introduce the functional design principles of ratiometric PA imaging probes based on core-shell nanoprobes, core-satellite nanoprobes, and universal hybrid nanoprobes, where we have established that reference signal and sensing signal can be obtained from the random assortment of plasmonic components and organic semiconducting molecules using a phase separation strategy. On these insights, we discuss the rational and detailed biomedical applications of ratiometric PA imaging probes which include accurate quantitative detection of disease-related molecules in inflammation or tumors in real time. In these champion implementations of ratiometric PA imaging probes, different diagnostic modules have been linked through compound modification with activation characteristics (e.g., pH, redox, enzyme, hypoxia). Finally, we present the challenges and perspectives for ratiometric probes based on optical imaging and PA imaging for multitarget design and future clinical translation. We believe that the upcoming generations of ratiometric imaging probes would have promising potential applications in the precise diagnosis of diseases. Finally, this Account may stimulate innovative studies in the design of ratiometric imaging probes and exploration of their clinical applications.