The Lethal and Sublethal Effects of Lambda-Cyhalothrin and Emamectin Benzoate on the Soybean Pest Riptortus pedestris (Fabricius).
Riptortus pedestris (Fabricius, 1775) (Hemiptera: Alydidae) is a major soybean pest in East Asia that can cause soybean staygreen syndrome. To date, no insecticides have been registered for the control of R. pedestris in China, and these insects are primarily controlled in the field through the application of broad-spectrum insecticides including lambda-cyhalothrin (LCT) and emamectin benzoate (EMB). Here, the lethal and sublethal effects of LCT and EMB on R. pedestris were comprehensively evaluated. LCT and EMB were both found to exhibit high levels of toxicity and concentration-dependent repellent effects for R. pedestris. The exposure of third instar nymphs from the F0 generation to LC30 concentrations of LCT and EMB resulted in a significant increase in the duration of nymph development and adult pre-oviposition period (APOP), together with reductions in fifth instar nymph and adult body weight, longevity, oviposition days, fecundity, vitellarium length, lateral oviduct diameter, and vitellogenin (Vg) gene expression as compared to control treatment. Strikingly, these suppressive effects were transmitted to the F1 generation, which similarly experienced the prolongation of preadult development and the preoviposition period (TPOP). Relative to control-treated populations, the F1 generation for these insecticide-treated groups also exhibited significant decreases in population parameter values. Overall, these data offer new insight into the impact that LCT and EMB treatment can have on R. pedestris, providing a valuable foundation for the application of these pesticides in the context of integrated pest management strategies aimed at soybean crop preservation.


Exopolysaccharides produced by Streptococcus mutans glucosyltransferases modulate the establishment of microcolonies within multispecies biofilms.
Streptococcus mutans is a key contributor to the formation of the extracellular polysaccharide (EPS) matrix in dental biofilms. The exopolysaccharides, which are mostly glucans synthesized by streptococcal glucosyltransferases (Gtfs), provide binding sites that promote accumulation of microorganisms on the tooth surface and further establishment of pathogenic biofilms. This study explored (i) the role of S. mutans Gtfs in the development of the EPS matrix and microcolonies in biofilms, (ii) the influence of exopolysaccharides on formation of microcolonies, and (iii) establishment of S. mutans in a multispecies biofilm in vitro using a novel fluorescence labeling technique. Our data show that the ability of S. mutans strains defective in the gtfB gene or the gtfB and gtfC genes to form microcolonies on saliva-coated hydroxyapatite surfaces was markedly disrupted. However, deletion of both gtfB (associated with insoluble glucan synthesis) and gtfC (associated with insoluble and soluble glucan synthesis) is required for the maximum reduction in EPS matrix and biofilm formation. S. mutans grown with sucrose in the presence of Streptococcus oralis and Actinomyces naeslundii steadily formed exopolysaccharides, which allowed the initial clustering of bacterial cells and further development into highly structured microcolonies. Concomitantly, S. mutans became the major species in the mature biofilm. Neither the EPS matrix nor microcolonies were formed in the presence of glucose in the multispecies biofilm. Our data show that GtfB and GtfC are essential for establishment of the EPS matrix, but GtfB appears to be responsible for formation of microcolonies by S. mutans; these Gtf-mediated processes may enhance the competitiveness of S. mutans in the multispecies environment in biofilms on tooth surfaces.


Potassium secretion along the inner medullary collecting duct.
Potassium transport along the inner medullary collecting duct (IMCD) was evaluated by the microcatheterization technique in Charles River CD (cesarean derived) rats 7-9 days after sham operation (S) or uninephrectomy (UNPX). The fraction of filtered potassium (TF/P)K/In) as a function of IMCD length was analyzed by linear regression. In 13 S rats there was a significant correlation and slope (P less than 0.001) and (TF/P)K/In increased from 14% at the beginning of the IMCD to 25% in the urine. IMCD potassium secretion accounted for about half of the excreted potassium. In the UNPX rats a significant correlation and slope was also obtained (P less than 0.001); (TF/P)K/In at the beginning of IMCD was 24% and increased to 36% in the urine. No difference in slope was noted between the groups. There was a slightly greater absolute potassium secretion after UNPX (0.77 +/- 0.03 S vs. 0.93 +/- 0.04 mueq/min UNPX), but this did not account for most of the difference in potassium excretion noted. We conclude that net potassium secretion occurs along the IMCD in S and UNPX rats. After UNPX, there is significantly greater potassium delivery to the IMCD and a greater kaluresis. The increased kaluresis cannot be accounted for primarily by increased potassium secretion along the IMCD.


Subsyndromal posttraumatic stress disorder is associated with health and psychosocial difficulties in veterans of Operations Enduring Freedom and Iraqi Freedom.
This study examined health and psychosocial correlates of subsyndromal/partial posttraumatic stress disorder (PTSD) and full PTSD in veterans of Operations Enduring Freedom and Iraqi Freedom (OEF/OIF). Five hundred and fifty-seven OEF/OIF veterans in Connecticut completed measures of PTSD and health and psychosocial functioning. A total 22.3% of the sample met screening criteria for partial PTSD and 21.5% for full PTSD. Veterans with partial PTSD reported an intermediate level of impairment (e.g., health ratings, work problems, relationship problems) relative to veterans without PTSD and veterans with full PTSD. These results suggest that subsyndromal/partial PTSD is associated with significant health and psychosocial difficulties and underscore the importance of assessing for partial PTSD in OEF/OIF veterans.


Human motor evoked potential responses following spinal cord transection: an in vivo study.
Motor evoked potential (MEP) monitoring has been used increasingly in conjunction with somatosensory evoked potential monitoring to monitor neurological changes during complex spinal operations. No published report has demonstrated the effects of segmental spinal cord transection on MEP monitoring. The authors describe the case of an 11-year-old girl with lumbar myelomeningocele and worsening thoracolumbar scoliosis who underwent a T11-L5 fusion and spinal transection to prevent tethering. Intraoperative MEP and somatosensory evoked potential monitoring were performed, and the spinal cord was transected in 4 quadrants. The MEPs were lost unilaterally as each anterior quadrant was sectioned. This is the first reported case that demonstrates the link between spinal cord transection and MEP signaling characteristics. Furthermore, it demonstrates the relatively minor input of the ipsilateral ventral corticospinal tract in MEP physiology at the thoracolumbar junction. Finally, this study further supports the use of MEPs as a specific intraoperative neuromonitoring tool.


Selection and allocation of manual traffic control points and personnel during emergencies.
Manual traffic control is an intersection control strategy in which law enforcement officers allocate intersection right-of-way to turning movements. Many emergency traffic management plans call for manual traffic control in response to oversaturated roadway conditions. This is because it is thought to more effectively move traffic during temporary surges in demand. The goal of this research was to evaluate the current state-of-the-practice used by the Army Corps of Engineers (ACE) in selecting intersections for manual traffic control and allocating police personnel to them during emergencies. This research uses the emergency traffic management plans developed by the ACE for nine counties in the Maryland Eastern Shore region. This area encompassing 14,318 intersections of which 74 were selected for manual traffic control during emergencies. This work sought to quantify the correlations that exist between intersection attributes and the ACE' decision to allocate officers to control them. The research findings suggest that US routes, State routes, and emergency evacuation routes are statistically significant in determining the need for police control at intersections. Also significant are intersection on contraflow corridors and intersections near grade separated interchanges. The model also determined that intersections isolated from evacuation routes and county exits were more likely to be selected for manual control, indicating that rural areas may rely on manual traffic control in the absence of multilane highway and freeways. This research also found that intersections involving evacuation routes, contraflow corridors, and grade separated interchanges may warrant additional police personnel (two or more officers) for manual traffic control.


18F-FET PET Uptake Characteristics in Patients with Newly Diagnosed and Untreated Brain Metastasis.
In patients with brain metastasis, PET using labeled amino acids has gained clinical importance, mainly regarding the differentiation of viable tumor tissue from treatment-related effects. However, there is still limited knowledge concerning the uptake characteristics in patients with newly diagnosed and untreated brain metastases. Hence, we evaluated the uptake characteristics in these patients using dynamic O-(2-18F-fluoroethyl)-l-tyrosine (18F-FET) PET. Methods: Patients with newly diagnosed brain metastases without prior local therapy and 18F-FET PET scanning were retrospectively identified in 2 centers. Static and dynamic PET parameters (maximal/mean tumor-to-brain-ratio [TBRmax/TBRmean], biologic tumor volume [BTV], and time-activity curves with minimal time to peak [TTPmin]) were evaluated and correlated with MRI parameters (maximal lesion diameter, volume of contrast enhancement) and originating primary tumor. Results: Forty-five brain metastases in 30 patients were included. Forty of 45 metastases (89%) had a TBRmax ≥ 1.6 and were classified as 18F-FET-positive (median TBRmax, 2.53 [range, 1.64-9.47]; TBRmean, 1.86 [range, 1.63-5.48]; and BTV, 3.59 mL [range, 0.04-23.98 mL], respectively). In 39 of 45 brain metastases eligible for dynamic analysis, a wide range of TTPmin was observed (median, 22.5 min; range, 4.5-47.5 min). All 18F-FET-negative metastases had a diameter of ≤ 1.0 cm, whereas metastases with a > 1.0 cm diameter all showed pathologic 18F-FET uptake, which did not correlate with lesion size. The highest variability of uptake intensity was observed within the group of melanoma metastases. Conclusion: Untreated metastases predominantly show increased 18F-FET uptake, and only a third of metastases < 1.0 cm were 18F-FET-negative, most likely because of scanner resolution and partial-volume effects. In metastases > 1.0 cm, 18F-FET uptake intensity was highly variable and independent of tumor size (even intraindividually). 18F-FET PET might provide additional information beyond the tumor extent by reflecting molecular features of a metastasis and might be a useful tool for future clinical applications, for example, response assessment.


Influence of starting material particle size on pellet surface roughness.
The purpose of this study was to investigate the effect of pelletization aids, i.e., microcrystalline cellulose (MCC) and cross-linked polyvinyl pyrrolidone (XPVP), and filler, i.e., lactose, particle size on the surface roughness of pellets. Pellets were prepared from powder blends containing pelletization aid/lactose in 1:3 ratio by extrusion-spheronization. Surface roughness of pellets was assessed quantitatively and qualitatively using optical interferometry and scanning electron microscopy, respectively. Both quantitative and qualitative surface studies showed that surface roughness of pellets depended on the particle size of XPVP and lactose used in the formulation. Increase in XPVP or lactose particle size resulted in rougher pellets. Formulations containing MCC produced pellets with smoother surfaces than those containing XPVP. Furthermore, surface roughness of the resultant pellets did not appear to depend on MCC particle size. Starting material particle size was found to be a critical factor for determining the surface roughness of pellets produced by extrusion-spheronization. Smaller particles can pack well with lower peaks and valleys, resulting in pellets with smoother surfaces. Similar surface roughness of pellets containing different MCC grades could be due to the deaggregation of MCC particles into smaller subunits with more or less similar sizes during wet processing. Hence, for starting materials that deaggregate during the wet processing, pellet surface roughness is influenced by the particle size of the material upon deaggregation.


Timing of DNA damage responses impacts persistence to fluoroquinolones.
Bacterial persisters are subpopulations of phenotypic variants in isogenic cultures that can survive lethal doses of antibiotics. Their tolerances are often attributed to reduced activities of antibiotic targets, which limit corruption and damage in persisters compared with bacteria that die from treatment. However, that model does not hold for nongrowing populations treated with ofloxacin, a fluoroquinolone, where antibiotic-induced damage is comparable between cells that live and those that die. To understand how those persisters achieve this feat, we employed a genetic system that uses orthogonal control of MazF and MazE, a toxin and its cognate antitoxin, to generate model persisters that are uniformly tolerant to ofloxacin. Despite this complete tolerance, MazF model persisters required the same DNA repair machinery (RecA, RecB, and SOS induction) to survive ofloxacin treatment as their nongrowing, WT counterparts and exhibited similar indicators of DNA damage from treatment. Further investigation revealed that, following treatment, the timing of DNA repair was critical to MazF persister survival because, when repair was delayed until after growth and DNA synthesis resumed, survival was compromised. In addition, we found that, with nongrowing, WT planktonic and biofilm populations, stalling the resumption of growth and DNA synthesis after the conclusion of fluoroquinolone treatment with a prevalent type of stress at infection sites (nutrient limitation) led to near complete survival. These findings illustrate that the timing of events, such as DNA repair, following fluoroquinolone treatment is important to persister survival and provide further evidence that knowledge of the postantibiotic recovery period is critical to understanding persistence phenotypes.


New Concerns for Neurocognitive Function during Deep Space Exposures to Chronic, Low Dose-Rate, Neutron Radiation.
As NASA prepares for a mission to Mars, concerns regarding the health risks associated with deep space radiation exposure have emerged. Until now, the impacts of such exposures have only been studied in animals after acute exposures, using dose rates ∼1.5×105 higher than those actually encountered in space. Using a new, low dose-rate neutron irradiation facility, we have uncovered that realistic, low dose-rate exposures produce serious neurocognitive complications associated with impaired neurotransmission. Chronic (6 month) low-dose (18 cGy) and dose rate (1 mGy/d) exposures of mice to a mixed field of neutrons and photons result in diminished hippocampal neuronal excitability and disrupted hippocampal and cortical long-term potentiation. Furthermore, mice displayed severe impairments in learning and memory, and the emergence of distress behaviors. Behavioral analyses showed an alarming increase in risk associated with these realistic simulations, revealing for the first time, some unexpected potential problems associated with deep space travel on all levels of neurological function.


Professional strategies in work-related practice: an exploration of occupational and physical therapy roles and approaches.
Both occupational and physical therapy have historical roots in worker rehabilitation; however, the core philosophies and areas of expertise of these professions suggest there may be inherent differences in the ways each approaches therapeutic intervention. This study surveyed 600 occupational and physical therapists in the United States to determine the nature and scope of work-related practice, and the degree to which occupation-based strategies are used by either profession. The overall response rate was 54% (n = 324), and 76% of respondents (n = 246) were actively engaged in providing work-related services. Results indicate that both professions provide all services commonly associated with work-related therapy in almost equal proportions. Few significant differences were identified between the professions in terms of approaches to therapy, and findings did not reveal the use of occupation-based interventions to any greater degree by occupational therapists.


Characterization of the gamma-interferon-mediated induction of antigen-presenting ability in P388D1 cells.
We characterized an assay system to study the lymphokine-mediated induction of antigen-presenting ability in P388D1 cells. The ability of lymphokine-induced P388D1 macrophages to present antigen plus Id was measured by their ability to induce interleukin 2 production by antigen-specific, Id-restricted T cell hybridomas in the presence of the appropriate antigen. The production of IL 2 by the T cell hybridomas is known to be dependent on the expression of Ia antigens by the antigen-presenting cells. The results obtained suggest that a factor present in the supernatant of the T cell hybridoma FS7-20.6.18 is responsible for inducing the appearance of I-Ad and I-Ed on P388D1, measured by immunofluorescence, and the ability of the cell to present antigen in association with I-Ad or I-Ed. The factor mediating the induction of antigen-presenting ability is thought to be gamma-interferon, because the hybridoma FS7-20.6.18 is known to produce this lymphokine and the factor is sensitive to pH 2 incubation. gamma-Interferon produced by recombinant DNA technology was found to induce antigen-presenting ability in this assay; however, alpha- and beta-interferon were inactive. This observation suggests a unique immunoregulatory role for gamma-interferon. Using many T cell hybridomas in the assay, we were able to distinguish three groups: a) high avidity hybridomas that respond to antigen presented by uninduced P388D1 but show an enhanced response to antigen plus induced P388D1; b) medium avidity hybridomas that do not respond to antigen presented by uninduced P388D1; and c) low avidity hybridomas that show a limited response to antigen presented by induced P388D1, but the response of which increases if the P388D1 cells are induced for longer periods of time. These different patterns of response are believed to be dependent on the Ia antigen density expressed by the gamma-interferon-induced presenting cells, and suggest that the T cell receptors for Ag/Id display marked heterogeneity in their avidities for Ag/Id.


Characterization and purification of osteogenic cells from murine bone marrow by two-color cell sorting using anti-Sca-1 monoclonal antibody and wheat germ agglutinin.
Osteogenic cells were sorted from bone marrow of 5-fluorouracil (5-FU)-treated mice based on light scatter characteristics, Sca-1 expression, and their binding to wheat germ agglutinin (WGA). Four sort gates were established using forward (FSC) and perpendicular (SSC) light scatter and were denominated as FSChigh SSClow, FSClow SSChigh, FSClow SSClow, and FSChigh SSChigh cell. Cells from the FSChigh SSChigh gate, but not from the other gates, synthesized alkaline phosphatase, collagen, and osteocalcin and formed a mineralized matrix in culture. The number of osteoprogenitor cells was significantly enriched after depleting the 5-FU bone marrow from cells of the lymphoid and myeloid lineage, eg, T cells, B cells, natural killer cells, granulocytes, macrophages, and erythrocytes. Approximately 95% of the FSChigh SSChigh cell population of this "lineage-negative" (Lin-) marrow expressed the Sca-1 antigen (Sca-1+) and bound WGA. Three additional sort windows were established based on WGA binding intensity and were denominated as Sca-1+ WGAdull, Sca-1+ WGAmedium, and Sca-1+ WGAbright. Cells from the Sca-1+ WGAbright gate, but not from the other gates, synthesized bone proteins and formed a mineralized matrix. However, they lost this capacity upon subcultivation. Further immunophenotypic characterization showed that FSChigh SSChigh Lin- Sca-1+ WGAbright cells expressed stromal (KM16) and endothelial (Sab-1 and Sab-2) markers, but not hematopoietic surface markers such as c-kit and Thy1.2. Sorted FSChigh SSChigh Lin- Sca-1+ WGAbright cells form three-dimensional nodules that stain with the von Kossa technique and contain osteoblast and osteocyte-like cells.


Desmoglein 2 is less important than desmoglein 3 for keratinocyte cohesion.
Desmosomes provide intercellular adhesive strength required for integrity of epithelial and some non-epithelial tissues. Within the epidermis, the cadherin-type adhesion molecules desmoglein (Dsg) 1-4 and desmocollin (Dsc) 1-3 build the adhesive core of desmosomes. In keratinocytes, several isoforms of these proteins are co-expressed. However, the contribution of specific isoforms to overall cell cohesion is unclear. Therefore, in this study we investigated the roles of Dsg2 and Dsg3, the latter of which is known to be essential for keratinocyte adhesion based on its autoantibody-induced loss of function in the autoimmune blistering skin disease pemphigus vulgaris (PV). The pathogenic PV antibody AK23, targeting the Dsg3 adhesive domain, led to profound loss of cell cohesion in human keratinocytes as revealed by the dispase-based dissociation assays. In contrast, an antibody against Dsg2 had no effect on cell cohesion although the Dsg2 antibody was demonstrated to interfere with Dsg2 transinteraction by single molecule atomic force microscopy and was effective to reduce cell cohesion in intestinal epithelial Caco-2 cells which express Dsg2 as the only Dsg isoform. To substantiate these findings, siRNA-mediated silencing of Dsg2 or Dsg3 was performed in keratinocytes. In contrast to Dsg3-depleted cells, Dsg2 knockdown reduced cell cohesion only under conditions of increased shear. These experiments indicate that specific desmosomal cadherins contribute differently to keratinocyte cohesion and that Dsg2 compared to Dsg3 is less important in this context.


Positive regulation of adult bone formation by osteoblast-specific transcription factor osterix.
Osterix (Osx) is essential for osteoblast differentiation and bone formation, because mice lacking Osx die within 1 h of birth with a complete absence of intramembranous and endochondral bone formation. Perinatal lethality caused by the disruption of the Osx gene prevents studies of the role of Osx in bones that are growing or already formed. Here, the function of Osx was examined in adult bones using the time- and site-specific Cre/loxP system. Osx was inactivated in all osteoblasts by Col1a1-Cre with the activity of Cre recombinase under the control of the 2.3-kb collagen promoter. Even though no bone defects were observed in newborn mice, Osx inactivation with 2.3-kb Col1a1-Cre exhibited osteopenia phenotypes in growing mice. BMD and bone-forming rate were decreased in lumbar vertebra, and the cortical bone of the long bones was thinner and more porous with reduced bone length. The trabecular bones were increased, but they were immature or premature. The expression of early marker genes for osteoblast differentiation such as Runx2, osteopontin, and alkaline phosphatase was markedly increased, but the late marker gene, osteocalcin, was decreased. However, no functional defects were found in osteoclasts. In summary, Osx inactivation in growing bones delayed osteoblast maturation, causing an accumulation of immature osteoblasts and reducing osteoblast function for bone formation, without apparent defects in bone resorption. These findings suggest a significant role of Osx in positively regulating osteoblast differentiation and bone formation in adult bone.


Merkel Cell Polyomavirus Antibody Titer Predicts Recurrence-Free Survival.
Merkel cell polyomavirus (MCPyV) is associated with the development of Merkel cell carcinoma (MCC). Antibody (MCPyV-Ab) titers may have prognostic implications. This study evaluated the impact of the presence or absence of MCPyV-Ab on the 2-year overall survival (OS) and disease-free survival (DFS) of MCC patients. This single-center, IRB-approved, retrospective cohort study evaluated 51 adult patients with MCC from 2014 to 2021 using a prospectively maintained database. Patients were compared by MCPyV-Ab status, and Kaplan-Meier analysis was used to evaluate 2-year OS and DFS. Of the 51 patients, 13 (25.4%) were seropositive, 41 (80.4%) underwent wide excision, 40 (80.0%) received radiotherapy, and 43 (84.3%) received multimodal therapy. The median follow-up period was 15.5 months (range 1-69.5 months). The median 2-year OS of the entire cohort was not reached. The median 2-year OS was not reached for either the seronegative or the seropositive patients. The difference in 2-year OS between the groups was not statistically significant (p = 0.37). Eight patients, all seronegative, were never rendered disease-free and were removed from recurrence analysis. The seropositive patients experienced no recurrences. Of the 30 seronegative patients, 9 (30.0%) experienced recurrence. The median 2-year DFS of the entire cohort was not reached. The median 2-year DFS of the seronegative group was 22.2 months. The 2-year DFS was not reached for the seropositive cohort. Seropositivity conferred a significantly better 2-year DFS than seronegativity (p = 0.04). The MCPyV-Ab seropositive patients demonstrated improved 2-year DFS. The seropositive patients showed a strong trend toward improved 2-year OS, although the difference not statistically significant. This study substantiated the value of MCPyV-Ab assessment for MCC.


Lifting capacity prediction model using physical performance measures among construction workers.
Manual materials handling is performed in many workplaces and is a significant risk factor for musculoskeletal injuries. The identification of lifting capacity is important to reduce the occurrence of musculoskeletal injuries. Lifting capacity is difficult to evaluate at the workplace. Therefore, there is a need to develop an alternate method that is easy and could be performed at the workplace. The study aimed to develop a lifting capacity prediction model for construction workers based on muscle strength and endurance. In this study, 65 construction workers were recruited; their socio-demographic and physical characteristics like core strength and endurance, grip strength, and lower limb flexibility were assessed. The lifting capacity was assessed using progressive isoinertial lifting evaluation. Stepwise multiple linear regression was carried out to develop the prediction model. The study suggested that age, BMI, grip strength, flexibility, prone plank, and trunk lateral flexor endurance tests have significantly influenced lifting capacity. Hence prediction model is developed using these variables. The regression model developed would help in easy estimation of lifting capacity among construction workers, which could be even administered with minimal skills by site supervisors or managers. It might help in the decision-making during pre-placement or return to work evaluations, thereby minimizing the incidence of low back disorders.


Information extraction from sound for medical telemonitoring.
Today, the growth of the aging population in Europe needs an increasing number of health care professionals and facilities for aged persons. Medical telemonitoring at home (and, more generally, telemedicine) improves the patient's comfort and reduces hospitalization costs. Using sound surveillance as an alternative solution to video telemonitoring, this paper deals with the detection and classification of alarming sounds in a noisy environment. The proposed sound analysis system can detect distress or everyday sounds everywhere in the monitored apartment, and is connected to classical medical telemonitoring sensors through a data fusion process. The sound analysis system is divided in two stages: sound detection and classification. The first analysis stage (sound detection) must extract significant sounds from a continuous signal flow. A new detection algorithm based on discrete wavelet transform is proposed in this paper, which leads to accurate results when applied to nonstationary signals (such as impulsive sounds). The algorithm presented in this paper was evaluated in a noisy environment and is favorably compared to the state of the art algorithms in the field. The second stage of the system is sound classification, which uses a statistical approach to identify unknown sounds. A statistical study was done to find out the most discriminant acoustical parameters in the input of the classification module. New wavelet based parameters, better adapted to noise, are proposed in this paper. The telemonitoring system validation is presented through various real and simulated test sets. The global sound based system leads to a 3% missed alarm rate and could be fused with other medical sensors to improve performance.


A practical guide to prescribing hormone replacement therapy.
Over the past 20 years there has been increasing interest in the menopause and hormone replacement therapy (HRT). More recently, postmenopausal HRT has been seen as a specific treatment for symptoms in the short term and preventative therapy in the long term. Women must be counselled regarding the risks and benefits of HRT according to the best available evidence. The patient should also be actively involved in the decision regarding HRT therapy, which should then improve patient compliance. Generally, an appropriate regimen of HRT can be formulated for the majority of patients. Progestogen should be added to therapy in women with an intact uterus in a cyclical or continuous regimen. The management of common estrogenic and progestogenic adverse effects is important in improving compliance. At present, new drugs are being developed for the management of the menopause (selective estrogen receptor modulators and phytoestrogens). Obviously, further research will be necessary to determine whether these drugs have advantages over regular HRT. By offering postmenopausal women HRT an attempt is made to optimise their physical and psychological well-being. However, HRT is not without adverse effects, the most worrying of which is the possible increase in breast cancer risk with long term use. However, with patient education efforts, treatment regimens acceptable to both patient and practitioner can be initiated; in this regard, the aim of the practitioner should be to help the menopausal woman make the decision which is the most appropriate for her.


Reconsidering fear of birth: Language matters.
Interest in fear of childbirth has grown exponentially since the 1980s, but the landscape of birth has shifted considerably since then, with evolving feminism; moving from a patriarchal environment in a biomedical model of care to a holistic model which recognizes the birth and sexuality rights of women and birthing people. Distinguishing the spectrum of fear from low to high and severe is important rather than aggregating all individuals with fear of childbirth. However, the terms 'fear of childbirth' and 'tocophobia' have been used interchangeably. In this paper we urge clinicians to use the term 'tocophobia' with caution since it may be construed negatively and there is a limited understanding of the underpinning aetiology of tocophobia. Furthermore, using the label may be disempowering for women and birthing people making decisions about their birth. Further research is warranted to better understand the experience, refine and define the issue and meet the individual needs of people with fear of childbirth and tocophobia.


Dual RNA Sequencing Reveals Key Events When Different Giardia Life Cycle Stages Interact With Human Intestinal Epithelial Cells In Vitro.
Giardia intestinalis is a protozoan parasite causing diarrheal disease, giardiasis, after extracellular infection of humans and other mammals' intestinal epithelial cells (IECs) of the upper small intestine. The parasite has two main life cycle stages: replicative trophozoites and transmissive cysts. Differentiating parasites (encysting cells) and trophozoites have recently been shown to be present in the same regions of the upper small intestine, whereas most mature cysts are found further down in the intestinal system. To learn more about host-parasite interactions during Giardia infections, we used an in vitro model of the parasite's interaction with host IECs (differentiated Caco-2 cells) and Giardia WB trophozoites, early encysting cells (7 h), and cysts. Dual RNA sequencing (Dual RNAseq) was used to identify differentially expressed genes (DEGs) in both Giardia and the IECs, which might relate to establishing infection and disease induction. In the human cells, the largest gene expression changes were found in immune and MAPK signaling, transcriptional regulation, apoptosis, cholesterol metabolism and oxidative stress. The different life cycle stages of Giardia induced a core of similar DEGs but at different levels and there are many life cycle stage-specific DEGs. The metabolic protein PCK1, the transcription factors HES7, HEY1 and JUN, the peptide hormone CCK and the mucins MUC2 and MUC5A are up-regulated in the IECs by trophozoites but not cysts. Cysts specifically induce the chemokines CCL4L2, CCL5 and CXCL5, the signaling protein TRKA and the anti-bacterial protein WFDC12. The parasite, in turn, up-regulated a large number of hypothetical genes, high cysteine membrane proteins (HCMPs) and oxidative stress response genes. Early encysting cells have unique DEGs compared to trophozoites (e.g. several uniquely up-regulated HCMPs) and interaction of these cells with IECs affected the encystation process. Our data show that different life cycle stages of Giardia induce different gene expression responses in the host cells and that the IECs in turn differentially affect the gene expression in trophozoites and early encysting cells. This life cycle stage-specific host-parasite cross-talk is an important aspect to consider during further studies of Giardia's molecular pathogenesis.


Oral manifestations related to immunosuppression degree in HIV-positive children.
Oral manifestations often found in HIV-infected children are frequently the first clinical sign of the infection. This article aims to report the prevalence of oral manifestations in soft tissues and their relationship with the degree of immunosuppression in 80 HIV-infected patients (average age 6.30 +/- 3.32 years old) at the IPPMG-UFRJ. Thirty children (38%) presented some type of oral lesion and the percentage of CD4 was lower than that found in lesion-free children (p < 0.05); 22.5% presented candidiasis, 17.5% gingivitis, 8.8% enlargement of parotids, 1.3% herpes simplex and 1.3% hairy leukoplakia. Of the 30 children with lesions, 70% showed severe immunosuppression, 23.3% moderate immunosuppression and in only 6.7% was immunosuppression absent. Oral manifestations were directly related to the degree of immunosuppression and such lesions can be considered as indicators of the progression of the HIV infection in children.


Recent advances in SPME techniques in biomedical analysis.
Biomedical analyses of drugs, metabolites, poisons, environmental and occupational pollutants, disease biomarkers and endogenous substances in body fluids and tissues are important in the development of new drugs, therapeutic monitoring, forensic toxicology, patient diagnosis, and biomonitoring of human exposure to hazardous chemicals. In these analyses, sample preparation is essential for isolation of desired components from complex biological matrices and greatly influences their reliable and accurate determination. Solid-phase microextraction (SPME) is an effective sample preparation technique that has enabled miniaturization, automation and high-throughput performance. The use of SPME has reduced assay times, as well as the costs of solvents and disposal. This review focuses on recent advances in novel SPME techniques, including fiber SPME and in-tube SPME, in biomedical analysis. We also summarize the applications of these techniques to pharmacotherapeutic, forensic, and diagnostic studies, and to determinations of environmental and occupational exposure.


Sequevar Diversity and Virulence of Ralstonia solanacearum Phylotype I on Mayotte Island (Indian Ocean).
The genetic and phenotypic diversity of the Ralstonia solanacearum species complex, which causes bacterial wilt to Solanacae, was assessed in 140 strains sampled from the main vegetable production areas of the Mayotte island. Only phylotype I strains were identified in the five surveyed areas. The strains were distributed into the following 4 sequevars: I-31 (85.7%), I-18 (5.0%), I-15 (5.7%), and I-46 (3.6%). The central area of Mayotte was the most diverse region, harboring 4 sequevars representing 47.1% of the collected strains. Virulence tests were performed under field and controlled conditions on a set of 10 tomato breeding line accessions and two commercial hybrid tomato cultivars. The strains belonging to sequevar I-31 showed the highest virulence on the tomatoes (pathotypes T-2 and T-3), whereas sequevars I-18, I-15, and I-46 were grouped into the weakly T-1 pathotype. When the tomato accessions were challenged in the field and growth chambers, the highest level of resistance were observed from the genetically related accessions Hawaii 7996, R3034, TML46, and CLN1463. These accessions were considered moderately to highly resistant to representative strains of the most virulent and prevalent sequevar (I-31). Interestingly, the Platinum F1 cultivar, which was recently commercialized in Mayotte for bacterial wilt resistance, was highly or moderately resistant to all strains. This study represents the first step in the rationalization of resistance deployment strategies against bacterial wilt-causing strains in Mayotte.


Epidermal growth factor receptor glycosylation is required for ganglioside GM3 binding and GM3-mediated suppression [correction of suppresion] of activation.
Gangliosides are able to bind to the epidermal growth factor receptor and inhibit its activation, but the mechanism of this inhibition is unknown. To address the role of receptor carbohydrates in facilitating interaction with gangliosides, we examined the ability of GM3 to bind the deglycosylated receptor and inhibit its autophosphorylation. Flow cytometry studies demonstrated that deglycosylation of the receptor did not affect its ability to be transported to the cell membrane. In contrast with the native (fully glycosylated) receptor, GM3 did not coimmunoprecipitate with the deglycosylated receptor. Using a novel colorimetric bead binding assay, GM3 was shown to bind well to the immunoprecipitated native receptor but not at all to the deglycosylated receptor. Finally, the addition of GM3 to cells with deglycosylated epidermal growth factor receptors did not result in significant further inhibition of autophosphorylation of the receptor, despite a 10-fold decrease in phosphorylation of the native epidermal growth factor receptor by 200 microM GM3. These studies suggest that ganglioside affects epidermal growth factor receptor activity through a direct interaction that requires receptor glycosylation, and contribute to our understanding of the role of gangliosides in cell membrane function.


SSRs and INDELs mined from the sunflower EST database: abundance, polymorphisms, and cross-taxa utility.
Simple sequence repeats (SSRs) are abundant and frequently highly polymorphic in transcribed sequences and widely targeted for marker development in eukaryotes. Sunflower (Helianthus annuus) transcript assemblies were built and mined to identify SSRs and insertions-deletions (INDELs) for marker development, comparative mapping, and other genomics applications in sunflower. We describe the spectrum and frequency of SSRs identified in the sunflower EST database, a catalog of 16,643 EST-SSRs, a collection of 484 EST-SSR and 43 EST-INDEL markers developed from common sunflower ESTs, polymorphisms of the markers among the parents of several intraspecific and interspecific mapping populations, and the transferability of the markers to closely and distantly related species in the Compositae. Of 17,904 unigenes in the transcript assembly, 1,956 (10.9%) harbored one or more SSRs with repeat counts of n > or = 5. EST-SSR markers were 1.6-fold more polymorphic among exotic than elite genotypes and 0.7-fold less polymorphic than non-genic SSR markers. Of 466 EST-SSR or INDEL markers screened for cross-species amplification and polymorphisms, 413 (88.6%) amplified alleles from one or more wild species (H. argophyllus, H. tuberosus, H. anomalus, H. paradoxus, and H. deserticola), whereas 69 (14.8%) amplified alleles from safflower (Carthamus tinctorius) and 67 (14.4%) amplified alleles from lettuce (Lactuca sativa); hence, only a fraction were transferable to distantly related genera in the Compositae, whereas most were transferable to wild relatives of H. annuus. Several thousand additional SSRs were identified in the EST database and supply a wealth of templates for EST-SSR marker development in sunflower.


"Model" patients and the consequences of provider responses to vaccine hesitancy.
The manner in which providers respond to parental concerns about the recommended childhood vaccination schedule is an area of increasing interest in pediatrics and medical ethics. Like many aspects of vaccination policy and practice, evaluating the reactions of providers to parental vaccine hesitancy--including the potential dismissal of such families from their practices--requires an appreciation of the twin character of vaccination as both a public health program as well as an individual preventive intervention. Accordingly, the ethics of vaccination combine many aspects of traditional medical ethics, such as respect for patient autonomy, the primacy of informed consent, and concern for the doctor-patient relationship, with the relatively newer discourse of public health ethics, one that directs particular attention to the health of populations and the pursuit of social justice. When parents are hesitant about vaccination for their children, providers may face a significant challenge reconciling their commitment to the health of those children, their respect for the perspectives of parents, and their interest in the health of their other patients and their communities. The tensions and potential conflicts among these considerations help to explain why provider responses to vaccine hesitancy have emerged as a frequent topic of discussion among practitioners, public health advocates, and ethicists alike.


Carbonate Adsorption to Ferrihydrite: Competitive Interaction with Phosphate for Use in Soil Systems.
Carbonate (CO3) interacts with Fe-(hydr)oxide nanoparticles, affecting the availability and geochemical cycle of other important oxyanions in nature. Here, we studied the carbonate-phosphate interaction in closed systems with freshly prepared ferrihydrite (Fh), using batch experiments that cover a wide range of pH values, ionic strength, and CO3 and PO4 concentrations. The surface speciation of CO3 has been assessed by interpreting the ion competition with the Charge Distribution (CD) model, using CD coefficients derived from MO/DTF optimized geometries. Adsorption of CO3 occurs predominately via formation of bidentate inner-sphere complexes, either (≡FeO)2CO or (≡FeO)2CO··Na+. The latter complex is electrostatically promoted at high pH and in the presence of adsorbed PO4. Additionally, a minor complex is present at high CO3 loadings. The CD model, solely parametrized by measuring the pH-dependent PO4 adsorption as a function of the CO3 concentration, successfully predicts the CO3 adsorption to Fh in single-ion systems. The adsorption affinity of CO3 to Fh is higher than to goethite, particularly at high pH and CO3 loadings due to the enhanced formation (≡FeO)2CO··Na+. The PO4 adsorption isotherm in 0.5 M NaHCO3 can be well described, being relevant for assessing the reactive surface area of the natural oxide fraction with soil extractions and CD modeling. Additionally, we have evaluated the enhanced Fh solubility due to Fe(III)-CO3 complex formation and resolved a new species (Fe(CO3)2(OH)2 3-(aq)), which is dominant in closed systems at high pH. The measured solubility of our Fh agrees with the size-dependent solubility predicted using the surface Gibbs free energy of Fh.


Regulating Size and Charge of Liposomes in Microneedles to Enhance Intracellular Drug Delivery Efficiency in Skin for Psoriasis Therapy.
The stratum corneum (SC) and cell membrane are two major barriers that hinder the therapeutic outcomes of transdermal drug delivery for the treatment of skin diseases. While microneedles (MNs) can efficiently penetrate the SC to deliver nanomedicines, the optimization of physicochemical properties of nanomedicines in MNs to enhance their in vivo cellular delivery efficiency remains unclear. Here, how the size and surface charge of drug-loaded liposomes in MNs influence the retention time and cellular delivery in psoriatic skin is systematically investigated. The results indicate that while 100 nm negatively-charged liposomes in MNs show higher cellular uptake in vitro, 250 and 450 nm liposomes could enhance skin retention and the long-term in vivo cellular delivery efficiency of drugs. Moreover, 250 nm cationic liposomes with a stronger positive charge show an extraordinarily long skin retention time of 132 h and significantly higher in vivo cellular internalization. In the treatment study, dexamethasone (dex)-loaded cationic liposomes-integrated MNs show better therapeutic outcomes than dex-loaded anionic liposomes-integrated MNs in a psoriasis-like animal model. The design principles of liposomes in MN drug delivery systems explored in the study hold the potential for enhancing the therapeutic outcomes of psoriasis and are instrumental for successful translation.


Strong enrichment of atmospherically relevant organic ions at the aqueous interface: the role of ion pairing and cooperative effects.
Surface affinity, orientation and ion pairing are investigated in mixed and single solute systems of aqueous sodium hexanoate and hexylammonium chloride. The surface sensitive X-ray photoelectron spectroscopy technique has been used to acquire the experimental results, while the computational data have been calculated using molecular dynamics simulations. By comparing the single solute solutions with the mixed one, we observe a non-linear surface enrichment and reorientation of the organic ions with their alkyl chains pointing out of the aqueous surface. We ascribe this effect to ion paring between the charged functional groups on the respective organic ion and hydrophobic expulsion of the alkyl chains from the surface in combination with van der Waals interactions between the alkyl chains. These cooperative effects lead to a substantial surface enrichment of organic ions, with consequences for aerosol surface properties.


The anti-CD20 antibody rituximab augments the immunospecific therapeutic effectiveness of an anti-CD19 immunotoxin directed against human B-cell lymphoma.
The chimaeric anti-CD20 antibody rituximab (Rituxan) sensitises lymphoma cells to small molecule cytotoxic drugs and to protein toxins. We have explored the augmentive effect of rituximab on the anti-CD19 immunotoxin BU12-SAPORIN in a model of human lymphoma. Intact rituximab and its F(ab)2 derivative both augmented the immunospecific protein synthesis inhibitory effects of BU12-SAPORIN in a complement-independent manner. A combination of rituximab + BU12-SAPORIN completely abolished the proliferation of Ramos cells in vitro and also induced a significantly greater degree of apoptosis in these cells. Treatment with rituximab, BU12-SAPORIN or a combination of both induced poly(ADPribose) polymerase and caspase 3 cleavage, although this was always consistently greater in combination-treated cells. zVAD almost completely inhibited apoptosis in rituximab- or BU12-SAPORIN-treated cells but only partially in combination-treated cells. In severe combined immunodeficient (SCID)-Ramos mice the combination of rituximab + BU12-SAPORIN was significantly better therapeutically than either single agent. The immunological fidelity of the therapeutic effect because of combination treatment was demonstrated through the failure of rituximab to augment an irrelevant anti-CD7 immunotoxin. The therapeutic efficacy of rituximab and combination treatment was reduced in SCID-Ramos mice depleted of serum complement while natural killer cell depletion failed to show any convincing role for antibody-dependent cellular cytotoxicity. This study shows a clear therapeutic advantage from using rituximab to immunospecifically augment immunotoxin cytotoxicity warranting further investigation.


Trimethoprim potentiates valproic acid-induced neural tube defects (NTDs) in mice.
The antiepileptic drug valproic acid (VPA) may produce NTDs because of interference with folate metabolism. Therefore, the possible interactions of VPA with the dihydrofolate reductase inhibitor trimethoprim (TM) was investigated. The combination of TM with sulfamethoxazol is used for treatment of urinary infections, the most common complications of pregnancy. TM (80 and 160 mg/kg) was given i.p. and orally, 0.5 and 1 h, respectively, prior to valproic acid (VPA, 300 and 400 mg/kg, s.c.) in day 8 pregnant NMRI mice. Fetuses were examined for exencephaly, resorptions, and fetal weight retardation on day 18 of gestation. TM (160 mg/kg, i.p.) produced no exencephaly or embryolethality, but increased fetal weight. Administration of TM (80 mg/kg, i.p.) increased VPA-induced exencephaly and fetal weight retardation but not embryolethality. Exencephaly rates induced by VPA (300 and 400 mg/kg) were 4% and 12.9% and were increased by coadministration of TM to 22.7% and 42.5%, respectively (P < 0.01). Oral TM also increased VPA-induced exencephaly and fetal weight retardation but with lower potency than i.p. injection. The observed effects were not due to altered VPA pharmacokinetics. These results support the view that VPA-induced neural tube defects may be mediated via an interaction with folate metabolism, and advise against TM-use in VPA-treated epileptics during pregnancy.


Peptide stapling by late-stage Suzuki-Miyaura cross-coupling.
The development of peptide stapling techniques to stabilise α-helical secondary structure motifs of peptides led to the design of modulators of protein-protein interactions, which had been considered undruggable for a long time. We disclose a novel approach towards peptide stapling utilising macrocyclisation by late-stage Suzuki-Miyaura cross-coupling of bromotryptophan-containing peptides of the catenin-binding domain of axin. Optimisation of the linker length in order to find a compromise between both sufficient linker rigidity and flexibility resulted in a peptide with an increased α-helicity and enhanced binding affinity to its native binding partner β-catenin. An increased proteolytic stability against proteinase K has been demonstrated.


A recurrent cross-sectional qualitative study exploring how low-income mothers define snacks and reasons for offering snacks during infancy.
Despite rapid increases in snacking in recent decades, little is known about snacking during infancy. This study explored how low-income mothers define snacks and their reasons for offering snacks during infancy. A recurrent cross-sectional qualitative approach was used to identify themes from semi-structured interviews with low-income mothers when their infants were 6 and 12 months of age. A purposive sample of mothers (N = 15) was recruited from Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) offices and childcare centers serving low-income families in Rhode Island. Mothers also completed demographic and infant feeding questionnaires. Independent thematic analyses were conducted to identify themes from the 6 and 12 month interviews. Themes from the 6 month interviews for how mothers defined snacks were: snacks are consumed between meals, snacks are smaller portions, and snacks are sweet. Themes from the 12 month interviews also included snacks are consumed between meals and snacks are smaller portions with one additional theme: snacks do not include all food groups. Themes from the 6 month interviews for the reasons mothers offered snacks were: infants seemed hungry, infants showed interest, and snacks help manage behavior. Themes from 12 month interviews also included snacks help manage behavior with two additional themes: snacks expose infants to different flavors and snacks expose infants to different textures. Findings suggest that snacks are commonly offered during infancy and that mothers define snacks as smaller portions that help with hunger between meals. However, during early infancy mothers describe snacks as sweet, and across infancy report using snacks to manage behavior, underscoring the importance of providing parents with guidance on healthy snacking during the first year of life.


Miyagi Medical and Welfare Information Network: A Backup System for Patient Clinical Information after the Great East Japan Earthquake and Tsunami.
On March 11, 2011, the Great East Japan Earthquake and ensuing tsunami that hit the northeastern coastal region of Japan caused about 18,000 casualties and destroyed numerous buildings. Additionally, many medical facilities were damaged and patient medical records lost. In order to maintain patient clinical information, a prefectural medical network system, the Miyagi Medical and Welfare Information Network (MMWIN), began providing backup data storage services in 2013 for hospitals, clinics, pharmacies, and other care facilities as a precaution for upcoming disasters. This system also facilitates the sharing of clinical information trans-institutionally as long as patients provide consent for this. In the present study, we examined the development of the MMWIN and its efficiency during the 5 years from its launch, and identified general problems to maintain such a backup system. At the end of 2018, the system contained backup data from more than 11 million patients with more than 420 million data items; more than 900 facilities were MMWIN users, and the number of patients consenting to sharing their clinical information reached 90,000. The use of the system has become widespread and the accumulating data should be utilized for research in the future. Maintaining a balance between income and cost is critical to make this project independent from local government subsidies.


Multichannel Microfluidic Platform for Temporal-Spatial Investigation of Niche Roles of Pseudomonas aeruginosa and Escherichia coli within a Dual-Species Biofilm.
In natural or man-made environments, microorganisms exist predominantly as biofilms forming surface-associated bacterial communities embedded in extracellular polymeric substances (EPSs). Often, biofilm reactors used for endpoint and disruptive analyses of biofilm are not suitable for periodic observation of biofilm formation and development. In this study, a microfluidic device designed with multiple channels and a gradient generator was used for high-throughput analysis and real-time monitoring of dual-species biofilm formation and development. We compared the structural parameters of monospecies and dual-species biofilms containing Pseudomonas aeruginosa (expressing mCherry) and Escherichia coli (expressing green fluorescent protein [GFP]) to understand the interactions in the biofilm. The rate of biovolume increase of each species in monospecies biofilm (2.7 × 105 μm3) was higher than those in a dual-species biofilm (9.68 × 104 μm3); however, synergism was still observed in the dual-species biofilm due to overall increases in biovolume for both species. Synergism was also observed in a dual-species biofilm, where P. aeruginosa forms a "blanket" over E. coli, providing a physical barrier against shear stress in the environment. The microfluidic chip was useful for monitoring the dual-species biofilm in the microenvironment, indicating that different species in a multispecies biofilm exhibit different niches for the survival of the biofilm community. Finally, we demonstrated that the nucleic acids can be extracted from the dual-species biofilm in situ after biofilm imaging analysis. In addition, gene expression supported that the activation and suppression of different quorum sensing genes resulted in the different phenotype seen in the biofilm. This study showed that the integration of microfluidic device with microscopy analysis and molecular techniques could be a promising tool for studying biofilm structure and gene quantification and expression simultaneously. IMPORTANCE In natural or man-made environments, microorganisms exist predominantly as biofilms forming surface-associated bacterial communities embedded in extracellular polymeric substances (EPSs). Often, biofilm reactors used for endpoint and disruptive analyses of biofilm are not suitable for periodic observation of biofilm formation and development. Here, we demonstrate that a microfluidic device with multiple channels and a gradient generator can be useful for high-throughput analysis and real-time monitoring of dual-species biofilm formation and development. Our study revealed synergism in the dual-species biofilm, where P. aeruginosa forms a "blanket" over E. coli, providing a physical barrier against shear stress in the environment. Furthermore, different species in a multispecies biofilm exhibit different niches for the survival of the biofilm community. This study showed that the integration of microfluidic device with microscopy analysis and molecular techniques could be a promising tool for studying biofilm structure and gene quantification and expression simultaneously.


Elbow radiographic anatomy: measurement techniques and normative data.
An increase in elbow pathology in adolescents has paralleled an increase in sports participation. Evaluation and classification of these injuries is challenging because of limited information regarding normal anatomy. The purpose of this study was to evaluate normal radiographic anatomy in adolescents to establish parameters for diagnosing abnormal development. Established and new measurements were evaluated for reliability and variance based on age and sex. Three orthopaedic surgeons independently, and in a standardized fashion, evaluated the normal anteroposterior and lateral elbow radiographs of 178 adolescent and young adult subjects. Fourteen measurements were performed including radial neck-shaft angle, articular surface angle, articular surface morphologic assessment (subjective and objective evaluation of the patterns of ridges and sulci), among others. We performed a statistical analysis by age and sex for each measure and assessed for inter- and intraobserver reliability. The distal humerus articular surface was relatively flat in adolescence and became more contoured with age, as objectively demonstrated by increasing depth of the trochlear and trochleocapitellar sulci, and decreasing trochlear notch angle. Overall measurements were similar between males and females, with an increased carrying angle in females. There were several statistically significant differences based on age and sex; but these were small and unlikely to be clinically significant. Inter and intraobserver reliability were variable; some commonly utilized tools had poor reliability. Most commonly utilized radiographic measures were consistent between sexes, across the adolescent age group, and between adolescents and young adults. Several commonly used assessment tools show poor reliability.


Galanin exerts dual action on inositol-specific phospholipase C activity in isolated pancreatic islets.
The intracellular mechanism whereby the neuropeptide galanin inhibits insulin secretion is not establish, since the peptide affects several signal pathways, including intracellular messengers such as calcium and cyclic AMP. In this study, we have assessed the effect of galanin on the inositol-specific phospholipase C (iPLC) activity in isolated rat pancreatic islets. The iPLC activity was measured as the generation of inositol 1,4,5-trisphosphate and its metabolite inositol 1,3,4-trisphosphate from the hydrolysis of polyphosphoinositides. Inositol phosphates were measured by anion-exchange fast protein liquid chromatography (FPLC) analysis of extracts from islets prelabelled with myo-3H-inositol. Galanin (1 to 100 nM) significantly increased the glucose-induced (12 mM) accumulation of inositol 1,4,5-trisphosphate after 2 min, but this stimulation of iPLC activity was followed by a significant suppression after 15 min. In the absence of extracellular calcium, both the stimulatory and inhibitory effects of galanin on the iPLC activity vanished. We therefore conclude that galanin initially stimulates iPLC in a calcium-dependent manner, followed by a secondary inhibitory effect. The secondary inhibition of iPLC activity might contribute to the insulinostatic action of the neuropeptide.


Distinct cortical activity patterns in Parkinson's disease and essential tremor during a bimanual tapping task.
Parkinson's disease (PD) and essential tremor (ET) are neurodegenerative diseases characterized by movement deficits. Especially in PD, maintaining cyclic movement can be significantly disturbed due to pathological changes in the basal ganglia and the cerebellum. Providing external cues improves timing of these movements in PD and also affects ET. The aim of this study is to determine differences in cortical activation patterns in PD and ET patients during externally and internally cued movements. Eleven PD patients, twelve ET patients, OFF tremor suppressing medication, and nineteen age-matched healthy controls (HC) were included and asked to perform a bimanual tapping task at two predefined cue frequencies. The auditory cue, a metronome sound presented at 2 or 4 Hz, was alternately switched on and off every 30 s. Tapping at two different frequencies were used since it is expected that different brain networks are involved at different frequencies as has been shown in previous studies. Cortical activity was recorded using a 64-channel EEG cap. To establish the cortical activation pattern in each group, the task related power (TRP) was calculated for each subject. For inter-groups analysis, EEG electrodes for divided into 5 different areas. Inter-group analysis revealed significant differences in areas responsible for motor planning, organization and regulation and involved in initiation, maintenance, coordination and planning of complex sequences of movements. Within the area of the primary motor cortex the ET group showed a significantly lower TRP than the HC group. In the area responsible for combining somatosensory, auditory and visual information both patient groups had a higher TRP than the HC group. Different neurological networks are involved during cued and non-cued movements in ET, PD and HC. Distinct cortical activation patterns were revealed using task related power calculations. Different activation patterns were revealed during the 2 and 4 Hz tapping task indicating different strategies to execute movements at these rates. The results suggest that a including a cued/non-cued tapping task during clinical decision making could be a valuable tool in an objective diagnostic protocol.


Cricoid arch replacement in dogs. Further studies.
Published reports by Delahunty et al and Alonso et al described the feasibility of correcting subglottic defects with cricoid arch transplantation. Using normal dogs, satisfactory results, including airway patency and vocal cord function, were reported. This report describes a second series of dogs in which subglottic stenosis was first induced by an open procedure and was subsequently repaired with an allotransplanted cricoid arch. The resulting subglottic defect in the donor dogs was repaired by autogenous hyoid arch transposition. No immunosuppressive measures or stents were used in either group. The animals were followed for periods up to one year prior to histologic study. Satisfactory results were obtained in both groups.


Site-specific Conjugation of 6 DOTA Chelators to a CA19-9-targeting scFv-Fc Antibody for Imaging and Therapy.
Antibodies conjugated with diagnostic/therapeutic radionuclides are attractive options for inoperable cancers lacking accurate imaging methods and effective therapeutics, such as pancreatic cancer. Hence, we have produced an antibody radionuclide conjugate termed TE-1132 comprising a α-CA19-9 scFv-Fc that is site-specifically conjugated at each C-terminus to 3 DOTA chelators via a cysteine-containing peptide linker. The smaller scFv-Fc size facilitates diffusivity within solid tumors, whereas the chelator-to-antibody ratio of six enabled 177Lu-radiolabeled TE-1132 to exhibit high radioactivity up to 520 MBq/nmol. In mice bearing BxPC3 tumors, immuno-SPECT/CT imaging of [111In]In-TE-1132 and the biodistribution of [177Lu]Lu-TE-1132 showed selective tumor accumulation. Single and multiple doses of [177Lu]Lu-TE-1132 effectively inhibited the BxPC3 tumor growth and prolonged the survival of mice with no irreversible body weight loss or hematopoietic damage. The adequate pharmacokinetic parameters, prominent tumor accumulation, and efficacy with good safety in mice encourage the further investigation of theranostic TE-1132 for treating pancreatic cancer.


Ultrasensitive detection of non-amplified genomic DNA by nanoparticle-enhanced surface plasmon resonance imaging.
Technologies today available for the DNA detection rely on a combination of labeled probes hybridized to target sequences which are amplified by polymerase chain reaction (PCR). Direct detection methods that eliminate the requirement for both PCR and labeling steps could afford faster, cheaper and simpler devices for the analysis of small amounts of unamplified DNA. In this work we describe the results obtained in the ultrasensitive detection of non-amplified genomic DNA. We analyzed certified reference materials containing different amounts of genetically modified DNA by using a detection method which combines the nanoparticle-enhanced surface plasmon resonance imaging (SPRI) biosensing to the peptide nucleic acids (PNAs) improved selectivity and sensitivity in targeting complementary DNA sequences. The method allowed us to obtain a 41 zM sensitivity in targeting genomic DNA even in the presence of a large excess of non-complementary DNA.


[The relationship between depression and cerebrovascular disease: clinical and biological aspects].
Medical complications after stroke are an important problem not only for patients, but also for their families and the clinicians who take care of them, thus representing a major public health problem. Among medical conditions complicating stroke, in the last several years much efforts has been directed to determine the role of affective disorders. Although depression coexisting with stroke has been shown to increase levels of functional disability and reduce the effectiveness of rehabilitation, we still have much to learn about the clinical interface between such disorders. This review focuses on the data concerning the potential relationship between depression and cerebrovascular disease and the emerging insights, which may be relevant to provide directions for the development of novel research strategies on the pathogenesis and treatment of the interface between depression and cerebrovascular disease.


Impact of ectomycorrhizal colonization of hybrid poplar on the remediation of diesel-contaminated soil.
Infection by ectomycorrhizal (ECM) fungi may benefit hybrid poplar growing in contaminated soils by providing greater access to water and nutrients and possibly protecting the trees from direct contact with toxic contaminants. The objective of this research was to determine the effect of colonization of the ECM fungus Pisolithus tinctorius (Pers.) Coker & Couch on hybrid poplar fine root production, biomass and N and P uptake when grown in diesel-contaminated soil (5000 mg diesel fuel kg soil(-1)). Commercially available Mycogrow Tree Tabs were the source of inoculum. A minirhizotron camera was used to provide the data necessary for estimating fine root production. Colonization of hybrid poplar roots (P. deltoides x [P. laurifolia x P. nigra] cv. Walker) by P. tinctorius increased total fine root production in diesel-contaminated soil to 56.58 g m(-2) compared to 22.59 g m(-2) in the uncolonized, diesel-contaminated treatment. Hybrid poplar leaf N and P concentrations were significantly greater in the diesel-contaminated/ECM-colonized treatment compared to the diesel-contaminated/uncolonized treatment after 12 wk, while significantly less diesel fuel was recovered from the soil of the uncolonized treatment compared to the colonized treatment. Both planted treatments removed more contaminants from the soil than an unplanted control. Significantly greater concentrations of total petroleum hydrocarbons (TPH) were found sequestered in hybrid poplar root/fungal-sheath complexes from the colonized treatment compared to the roots of the uncolonized treatment. The results of this study indicate that over a 12-wk growth period, ECM colonization of hybrid poplar in diesel-contaminated soils increased fine root production and whole-plant biomass, but inhibited removal of TPH from the soil.


Participation of active-site carboxylates of Escherichia coli DNA polymerase I (Klenow fragment) in the formation of a prepolymerase ternary complex.
We have investigated the roles of four active-site carboxylates in the formation of a prepolymerase ternary complex of Escherichia coli DNA polymerase I (Klenow fragment), containing the template-primer and dNTP. The analysis of nine mutant enzymes with conserved and nonconserved substitutions of Asp(705), Glu(710), Asp(882), and Glu(883) clearly shows that both catalytically essential aspartates, Asp(705) and Asp(882), are required for the formation of a stable ternary complex. Of the two glutamates, only Glu(710) is required for ternary complex formation, while Glu(883) does not participate in this process. This investigation also reveals two interesting properties of the Klenow fragment with regard to enzyme-template-primer binary and enzyme-template-primer-dNTP ternary complex formation. These are (a) the significant resistance of enzyme-template-primer-dNTP ternary complexes to the addition of high salt or template-primer challenge and (b) the ability of the Klenow fragment to form ternary complexes in the presence of noncatalytic divalent cations such as Ca(2+), Co(2+), Ni(2+), and Zn(2+).


Angiotensin II stimulates sympathetic output by a direct spinal action.
When administered intrathecally in a dose of 10 mu to the ninth thoracic segment of the spinal cord in the anesthetized rat, angiotensin II produced a transient increase in systolic and diastolic arterial pressures lasting 1-4 min. Heart rate was also increased, but in this case for more than 30 min. Similar administration of 5 micrograms or of CSF had no effect on either arterial pressure or heart rate. Neither the arterial pressure nor the heart rate response to 10 micrograms of angiotensin II was observed in rats given hexamethonium (10 mg/kg, i.v.), suggesting that the effects were mediated by spinal activation of sympathetic output. When the rats were pretreated with 10 micrograms of [Sar1, IIe8]-angiotensin II three min prior to angiotensin II, there was a block of the increase in arterial pressure but not of the increase in heart rate. When the antagonist was given 15 min prior to angiotensin II, the full pressor response appeared, suggesting that the antagonist was effective for less than 15 min; in addition, after the antagonist alone, while arterial pressure remained unaltered, there was a gradual increase in heart rate suggesting that the analogue had agonistic effects on mechanisms regulating heart rate. These results suggest that angiotensin II activates sympathetic mechanisms by a spinal action and that arterial pressure and heart rate are regulated differentially, arterial pressure via a mechanism which is antagonized by [Sar1, IIe8]-angiotensin II, and heart rate via a mechanism in which the analog can act as an agonist.


Re-treatment with varenicline is a cost-effective aid for smoking cessation.
One quit attempt with varenicline has been found to be a cost-effective smoking cessation intervention. The purpose of this study was to analyze varenicline's cost-effectiveness in patients who relapse during or after the first treatment. A comparison was made between re-treatment schema with varenicline and re-treatment schema with bupropion, NRT and unaided cessation, and treatment once with varenicline in a Finnish context. The two-quit version of BENESCO Markov model was used to follow a cohort of smokers making up to two quit attempts over a lifetime. The abstinence rates of the interventions were derived from a Cochrane review. Gender- and age-specific data on the incidence and prevalence of five smoking-related diseases were included in the model. Quality-adjusted life-years, total expected costs, and the lifetime cumulative incidence of smoking-related morbidities and mortality were the primary outcomes evaluated. The study cohort comprised 116,533 smokers who were willing to make a quit attempt. In the lifetime simulation, re-treatment with varenicline yielded 6,150-20,250 extra quitters, depending on the comparator. Among these quitters it was possible to prevent 899-2,972 additional cases of smoking-related diseases, and 395-1,307 deaths attributable to smoking. Re-treatment with varenicline resulted in cost savings of up to 54.9 million Euros. Re-treatment with varenicline dominated all the other smoking cessation interventions used in the analysis. Sensitivity analysis supported the robustness of the base case results. The analysis did not consider adverse events, and included only five major smoking-related diseases, which is a conservative approach, and probably leads to under-estimation of cost-effectiveness of cessation interventions. Furthermore, assumptions of constant relative risks for smoking-related diseases for each smoking status and the proxy values used as efficacy estimates of second quit attempts for other interventions than varenicline are limitations. A second quitting effort with varenicline is economically justifiable.


Comparison of fluid filtration and bacterial leakage techniques for evaluation of microleakage in endodontics.
Apical leakage assessment is a way to compare the efficiency of a filling material to seal the apical region of the tooth. Many microleakage testing techniques have been introduced through the years, but there has been no agreement as to which technique gives the most accurate results. The aim of this study was to compare the accuracy of fluid filtration and bacterial leakage techniques in the assessment of the apical sealing ability of mineral trioxide aggregate (MTA) and calcium enriched mixture (CEM). A sample of 34 extracted single-rooted human teeth were selected and prepared. The samples were divided in to 2 experimental groups. The apical 3 mm of each root was resected at 90° to its long axis and root end preparation was done with ultrasonic tips to a depth of 3 mm and filled with MTA and CEM, respectively. Assessment of apical sealing ability was done with fluid filtration technique and bacterial leakage technique along 90 days with Enterococcus faecalis bacteria. Mann-Whitney U-test and Chi-square test were used to analyze the data using SPSS (SPSS Inc., Chicago, IL, USA). P less than 0.05 was considered as significant. There was no significant difference in apical sealing ability between MTA and CEM in bacterial leakage and fluid filtration techniques. Samples which had bacterial leakage showed higher leakage values by fluid filtration technique. Both techniques showed same results and there was no significant difference between fluid filtration and bacterial leakage techniques in assessment of apical microleakage.


Interactions between 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and hypoxia signaling pathways in zebrafish: hypoxia decreases responses to TCDD in zebrafish embryos.
The aryl hydrocarbon receptor (AHR) interacts with the aryl hydrocarbon receptor nuclear translocator (ARNT) to form a heterodimer that binds to promoters in target genes to alter transcription in response to xenobiotics such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The ARNT protein also forms heterodimers with other proteins such as HIF-1alpha and HIF-2alpha to alter gene expression in response to low oxygen conditions. Because ARNT is shared between multiple signaling pathways it is possible that activation of one ARNT-requiring pathway could inhibit the activation of other pathways that depend on ARNT. One hypothesis to explain TCDD toxicity in early life stage fish is that TCDD activation of zfAHR2 sequesters zfARNT2 away from the hypoxia signaling pathway. To test this hypothesis we measured the ability of TCDD to prevent induction of heme oxygenase by hypoxia (40% saturation), as well as the ability of hypoxia to increase the sensitivity of zebrafish to the effects of TCDD during the first week of life. As a further test of the model we examined mutant zebrafish that lack zfARNT2 for phenotypes that resemble the effects of TCDD exposure. Our results demonstrate that sequestration of zfARNT2 is not causing TCDD toxicity. TCDD did not inhibit hypoxia induction of heme oxygenase, hypoxia and TCDD exposures were not additive in causing developmental toxicity, and mutant embryos that lack zfARNT2 do not develop defects mimicking TCDD toxicity. However, our results demonstrate some level of cross talk between the two pathways in the zebrafish embryo. Hypoxia decreased TCDD induction of zfCYP1A mRNA, and decreased the potency of TCDD in causing edema. It is not clear whether this is mediated through competition for zfARNT2, or through other mechanisms.


Multiple cancers. Tumor burden permits the outgrowth of other cancers.
We demonstrate that tumor-bearing hosts permit the outgrowth of "potentially malignant" cells that are located at a different site. These second cancers continued to grow and kill their hosts even though they retain the "premalignant" phenotype, even after removal of the original malignancy. The potentially malignant cells used in these experiments were ultraviolet light- or methylcholanthrene-induced regressor tumor cells that are rejected regularly by normal mice at any testable dose, and only form progressive tumors in immunosuppressed individuals. The immunological rejection of these highly immunogenic, potentially malignant cells was suppressed by Thy-1+, Ly-2-, nonadherent, radio-sensitive suppressor cells in the tumor-bearing mice. These suppressor cells were absent in nude tumor-bearing mice. Unlike helper and cytolytic T cell-mediated responses, which are exquisitely tumor specific, the suppression caused by a progressively growing tumor was crossreactive among many syngeneic, independently derived tumors induced by different carcinogens. However, T cell-mediated immune responses to alloantigens, allogeneic tumors, certain syngeneic tumors, and humoral responses to xenogeneic red blood cells were normal in these mice. The immune suppression in the tumor-bearing animals closely simulated that induced by ultraviolet light irradiation, and both types of suppression might therefore share common mechanisms. Our findings may contribute to understanding the growth, development, and possible control of multicentric malignancies and add a precaution to the potential use of strongly immunogenic tumor variants for active immunotherapy in hosts bearing less immunogenic tumors.


[The action of tuftsin on the reaction of macrophage-suppressor formation in vitro and in vivo].
A cell suspension consisting of nonadhering and adhering spleen cells in the ratio 30:1 was incubated in a 10(-4) M tuftsin's solution during 15-30 min. The addition of 10(7) cells incubated in tuftsin syngeneic recipients resulted in the suppression of the immune response of the latter to sheep red blood cells. It was noted that this effect may be induced by using adhering cells only of intact donors and only when incubated together with nonadhering cells. The addition of tuftsin one hour after transplantation of the nonadhering spleen cells resulted in the suppression of the immune response. It was proposed, that suppression effect released through generation of the macrophage-suppressors.


A refractory phase in cyclic AMP-responsive transcription requires down regulation of protein kinase A.
Cyclic AMP (cAMP) stimulates the expression of numerous genes through the protein kinase A (PK-A)-mediated phosphorylation of the nuclear factor CREB at Ser-133 (G. A. Gonzalez and M. R. Montminy, Cell 59:675-680, 1989). Like other signal transduction pathways, cAMP induces gene expression with burst-attenuation kinetics; cAMP-dependent transcription and CREB phosphorylation peak within 30 min and decline steadily over the next 4 to 6 h via the protein phosphatase 1-mediated dephosphorylation of CREB (M. Hagiwara, A. Alberts, P. Brindle, J. Meinkoth, J. Feramisco, T. Deng, M. Karin, S. Shenolikar, and M. Montminy, Cell 70:105-113, 1992). Here we characterize a third phase in cAMP-responsive transcription--a refractory period during which hormone-treated cells become transcriptionally unresponsive to subsequent stimulation by cAMP. This refractory period begins 6 to 8 h after stimulation and lasts 3 to 5 days after the removal of hormone. In contrast to the earlier attenuation phase, transcription of cAMP-responsive genes during the refractory period is not restored by inhibitors of protein phosphatase 1 activity. Rather, the establishment and maintenance of this phase rely on a marked reduction in PK-A catalytic subunit expression at the translational level. As overexpression of C-subunit protein can reactive transcription of cAMP-responsive genes during the refractory period, our results suggest that hormone-responsive cells may stimulate, attenuate, and then silence signal-dependent genes through distinct regulatory mechanisms.


Clinical risk factors for malignancy and overall survival in patients with pheochromocytomas and sympathetic paragangliomas: primary tumor size and primary tumor location as prognostic indicators.
Pheochromocytomas and sympathetic paragangliomas are rare neuroendocrine tumors for which no precise histological or molecular markers have been identified to differentiate benign from malignant tumors. The aim was to determine whether primary tumor location and size are associated with malignancy and decreased survival. We performed a retrospective chart review of patients with either pheochromocytoma or sympathetic paraganglioma. The study group comprised 371 patients. Overall survival and disease-specific survival were analyzed according to tumor size and location. Sixty percent of patients with sympathetic paragangliomas and 25% of patients with pheochromocytomas had metastatic disease. Metastasis was more commonly associated with primary tumors located in the mediastinum (69%) and the infradiaphragmatic paraaortic area, including the organ of Zuckerkandl (66%). The primary tumor was larger in patients with metastases than in patients without metastatic disease (P < 0.0001). Patients with sympathetic paragangliomas had a shorter overall survival than patients with pheochromocytomas (P < 0.0001); increased tumor size was associated with shorter overall survival (P < 0.001). Patients with sympathetic paragangliomas were twice as likely to die of disease than patients with pheochromocytomas (hazard ratio = 1.93; 95% confidence interval = 1.20-3.12; P = 0.007). As per multivariate analysis, the location of the primary tumor was a stronger predictor of metastases than was the size of the primary tumor. The size and location of the primary tumor were significant clinical risk factors for metastasis and decreased overall survival duration. These findings delineate the follow-up and treatment for these tumors.


Adoption and impact of improved groundnut varieties on household food security in Nigeria.
Improved agricultural technologies are promoted as cost-effective and sustainable ways of improving rural households' food security and reducing poverty in developing countries. This study evaluates the relationship between improved groundnut varieties (IGVs) and household food security using detailed household and plot level data from a sample of over 1300 farm households in Northern Nigeria. Endogenous switching regression models are employed to control for potential endogeneity biases. Results show that about 30 % of groundnut plots are planted with improved varieties, and the adoption of IGVs significantly increases the likelihood of household per capita groundnut consumption by about 13 % and reduces the probability of households' vulnerability to food (access) insecurity by 22 %. Counterfactual analyses show that non-adopting households could have enjoyed comparable benefits had they adopted IGVs. These results suggest that development interventions aimed at improving the diffusion and impacts of IGVs in Nigeria need to target farmers' access to information about the technologies while developing groundnut seed systems to make quality seeds readily available to smallholder farmers at affordable prices.


Cutaneous squamous cell carcinoma in patients with chronic lymphocytic leukemia: a systematic review of the literature.
The continuous improvement of life expectancy of patients with chronic lymphocytic leukemia (CLL) has resulted in increased risk of second primary malignancy that potentially may affect survival and quality of life of CLL patients. We performed a systematic review to assess the risk and the clinical-pathological features and prognosis of cutaneous squamous cell carcinoma (cSCC) in patients with CLL. We searched PubMed, Embase, and Cochrane Central Register of Control Trials databases for articles published from database inception to December 31, 2019. English-language studies reporting original data on patients with a specific diagnosis of CLL and cSCC were included. Data were extracted using a standardized extraction form, and any discordance was resolved by consensus. Descriptive data were generated by pooling patients from eligible studies. Of the 4588 non-duplicate records identified, 55 articles met our inclusion criteria. These studies reported that CLL patients have a 3.2% prevalence of cSCC, with an 11.5% cSCC-related lethality and an overall risk of metastasis of 5.7% (7.3% for regional lymph node involvement and 3.8% for distant metastasis). The quality of evidence was limited by the high heterogeneity in the design, populations, and objectives of the included studies. This systematic review suggests that cSCC in CLL patients tends to behave less aggressively compared with the solid organ transplant recipients but has a higher morbidity and mortality than in the general population. Future prospective studies are needed to increase the quality of evidence and to determine the best treatment modalities and screening intervals for these patients.


A Comparison of Person-Fit Indices to Detect Social Desirability Bias.
Social desirability bias (SDB) has been a major concern in educational and psychological assessments when measuring latent variables because it has the potential to introduce measurement error and bias in assessments. Person-fit indices can detect bias in the form of misfitted response vectors. The objective of this study was to compare the performance of 14 person-fit indices to identify SDB in simulated responses. The area under the curve (AUC) of receiver operating characteristic (ROC) curve analysis was computed to evaluate the predictive power of these statistics. The findings showed that the agreement statistic (A) outperformed all other person-fit indices, while the disagreement statistic (D), dependability statistic (E), and the number of Guttman errors (G) also demonstrated high AUCs to detect SDB. Recommendations for practitioners to use these fit indices are provided.


Characterisation and molecular dynamic simulations of J15 asparaginase from Photobacterium sp. strain J15.
Genome mining revealed a 1011 nucleotide-long fragment encoding a type I L-asparaginase (J15 asparaginase) from the halo-tolerant Photobacterium sp. strain J15. The gene was overexpressed in pET-32b (+) vector in E. coli strain Rosetta-gami B (DE3) pLysS and purified using two-step chromatographic methods: Ni(2+)-Sepharose affinity chromatography and Q-Sepharose anion exchange chromatography. The final specific activity and yield of the enzyme achieved from these steps were 20 U/mg and 49.2%, respectively. The functional dimeric form of J15-asparaginase was characterised with a molecular weight of ~70 kDa. The optimum temperature and pH were 25°C and pH 7.0, respectively. This protein was stable in the presence of 1 mM Ni(2+) and Mg(2+), but it was inhibited by Mn(2+), Fe(3+) and Zn(2+) at the same concentration. J15 asparaginase actively hydrolysed its native substrate, l-asparagine, but had low activity towards l-glutamine. The melting temperature of J15 asparaginase was ~51°C, which was determined using denatured protein analysis of CD spectra. The Km, Kcat, Kcat/Km of J15 asparaginase were 0.76 mM, 3.2 s(-1), and 4.21 s(-1) mM(-1), respectively. Conformational changes of the J15 asparaginase 3D structure at different temperatures (25°C, 45°C, and 65°C) were analysed using Molecular Dynamic simulations. From the analysis, residues Tyr₂₄ , His₂₂, Gly₂₃, Val₂₅ and Pro₂₆ may be directly involved in the 'open' and 'closed' lid-loop conformation, facilitating the conversion of substrates during enzymatic reactions. The properties of J15 asparaginase, which can work at physiological pH and has low glutaminase activity, suggest that this could be a good candidate for reducing toxic effects during cancer treatment.


The functionalization of nanodiamonds (diamondoids) as a key parameter of their easily controlled self-assembly in micro- and nanocrystals from the vapor phase.
We detail herein readily accessible processes to control previously unobserved robust self-assemblies of nanodiamonds (diamondoids) in micro- and nanocrystals from their mild vapor deposition. The chemical functionalization of uniform and discernible nanodiamonds was found to be a key parameter, and depending on the type of functional group (hydroxy, fluorine, etc.) and its position on the diamondoid, the structure of the discrete deposits can vary dramatically. Thus, well-defined anisotropic structures such as rod, needle, triangle or truncated octahedron shapes can be obtained, and self-assembled edifices of sizes ranging from 20 nm to several hundred micrometers formed with conservation of a similar structure for a given diamondoid. Key thermodynamic data including sublimation enthalpy of diamondoid derivatives are reported, and the SEM of the self-assemblies coupled with EDX analyses and XRD attest the nature and purity of nanodiamond crystal deposits. This attractive method is simple and outperforms in terms of deposit quality dip-coating methods we used. This vapor phase deposition approach is expected to allow for an easy formation of diamondoid nanoobjects on different types of substrates.


CB1 receptor signaling regulates social anxiety and memory.
The endocannabinoid (eCB) system regulates emotion, stress, memory and cognition through the cannabinoid type 1 (CB1 ) receptor. To test the role of CB1 signaling in social anxiety and memory, we utilized a genetic knockout (KO) and a pharmacological approach. Specifically, we assessed the effects of a constitutive KO of CB1 receptors (CB1 KOs) and systemic administration of a CB1 antagonist (AM251; 5 mg/kg) on social anxiety in a social investigation paradigm and social memory in a social discrimination test. Results showed that when compared with wild-type (WT) and vehicle-treated animals, CB1 KOs and WT animals that received an acute dose of AM251 displayed anxiety-like behaviors toward a novel male conspecific. When compared with WT animals, KOs showed both active and passive defensive coping behaviors, i.e. elevated avoidance, freezing and risk-assessment behaviors, all consistent with an anxiety-like profile. Animals that received acute doses of AM251 also showed an anxiety-like profile when compared with vehicle-treated animals, yet did not show an active coping strategy, i.e. changes in risk-assessment behaviors. In the social discrimination test, CB1 KOs and animals that received the CB1 antagonist showed enhanced levels of social memory relative to their respective controls. These results clearly implicate CB1 receptors in the regulation of social anxiety, memory and arousal. The elevated arousal/anxiety resulting from either total CB1 deletion or an acute CB1 blockade may promote enhanced social discrimination/memory. These findings may emphasize the role of the eCB system in anxiety and memory to affect social behavior.


Health-related quality of life in patients on hemodialysis and peritoneal dialysis.
Chronic kidney disease (CKD) is a worldwide public health problem, and its treatment imposes a considerable burden on patients and their families. Limitations in everyday activity may worsen the situation and affect the health-related quality of life (HRQOL) of patients with CKD. There are no studies on the HRQOL of dialysis patients in South Africa. We assessed the HRQOL of patients undergoing hemodialysis (HD) and continuous ambulatory peritoneal dialysis (PD) attending the Groote Schuur Hospital renal unit by using the Kidney Disease Quality of Life-Short Form version 1.3 questionnaire. Baseline demographic and clinical details of the participants were recorded. Analysis was performed (unpaired t test and univariate analysis) to compare the HRQOL between HD and PD patients and to identify factors influencing HRQOL. The HRQOL was low but not significantly different between HD and PD patients. In PD patients, the use of erythropoiesis-stimulating agents (ESA) significantly contributed to the emotional well-being (r 2 = 0.267; P = 0.01) and alleviation of pain (r 2 = 0.073; P = 0.049); in HD patients also, ESA use was associated with emotional well-being (r 2 = 0.258; P <0.0001) as well as improvement in energy/fatigue (r 2 = 0.390; P <0.0001). Systolic and diastolic blood pressures significantly influenced cognitive function in PD patients (P <0.05). Parathyroid hormone level significantly influenced the physical functioning and energy/fatigue domains in HD patients (P <0.0001). Serum ferritin (r 2 = 0.441; P = 0.002) and level of hemoglobin concentration (r 2 = 0.180; P = 0.006) were significantly associated with the domain role emotional in PD and HD patients, respectively. Although HRQOL is low in dialysis patients in Cape Town, the factors that have been identified to be associated with these scores (such as anemia and hyperparathyroidism) if aggressively managed and corrected may assist in improving patients' HRQOL.


Purification and characterization of the membrane-bound ferrochelatase from Spirillum itersonii.
The membrane-bound enzyme ferrochelatase (protoheme ferro-lyase, EC 220.127.116.11) was purified from isolated membrane fragments of Spirillum itersonii approximately 490-fold. Purification was achieved by solubilization with chaotropic salts followed by ammonium sulfate fractionation, diethylaminoethyl-cellulose chromatography, and gel filtration on Sephadex G-200. The purified enzyme has an apparent minimum molecular weight of approximately 50,000, as determined by gel filtration in the presence of 0.1% Brij 35 and 1 mM dithiothreitol but forms high-molecular-weight aggregates in the absence of detergent. Purified ferrochelatase is strongly stimulated in the presence of copper. The apparent Km for Fe2+ is 20 micrometer in the absence of copper and 9.5 micrometer in the presence of 20 micrometer CuCl2. The apparent Km for protoporphyrin is 50 micrometer, and it is unaltered by copper. Ferrochelatase has a single pH optimum of 7.50, and it is inhibited 50% by 20 micrometer heme. Certain divalent cations and sulfhydryl reagents also inhibit the enzyme.


A Music Therapy Intervention for Refugee Children and Adolescents in Schools: A Process Evaluation Using a Mixed Method Design.
Refugee children and adolescents have often experienced negative or traumatic events, which are associated with stress and mental health problems. A specific music therapy intervention is developed for this group in school settings. The aim of the present study was to set the first steps in the implementation of this intervention. A process evaluation was performed using a mixed method design among refugee children and adolescents (6-17 years) at three different schools in the Netherlands. Interviews were conducted with teachers and music therapists before, at the midpoint, and after the intervention. At these moments, children completed a classroom climate questionnaire and a visual analogue scale on affect. The results indicate that the intervention strengthens the process of social connectedness, resulting in a "sense of belonging". The intervention may stimulate inclusiveness and cultural sensitivity, and may contribute to a safe environment and the ability of teachers to adapt to the specific needs of refugee children. Refugee children and adolescents showed a decrease of negative affect during the intervention. When implementing the intervention in schools, it is important to take into account the initial situation, the prerequisites for the intervention, the professional competence, the experience of music therapists, and the collaboration and communication between the professionals involved.