p53 and TAp63 promote keratinocyte proliferation and differentiation in breeding tubercles of the zebrafish.
p63 is a multi-isoform member of the p53 family of transcription factors. There is compelling genetic evidence that ΔNp63 isoforms are needed for keratinocyte proliferation and stemness in the developing vertebrate epidermis. However, the role of TAp63 isoforms is not fully understood, and TAp63 knockout mice display normal epidermal development. Here, we show that zebrafish mutants specifically lacking TAp63 isoforms, or p53, display compromised development of breeding tubercles, epidermal appendages which according to our analyses display more advanced stratification and keratinization than regular epidermis, including continuous desquamation and renewal of superficial cells by derivatives of basal keratinocytes. Defects are further enhanced in TAp63/p53 double mutants, pointing to partially redundant roles of the two related factors. Molecular analyses, treatments with chemical inhibitors and epistasis studies further reveal the existence of a linear TAp63/p53->Notch->caspase 3 pathway required both for enhanced proliferation of keratinocytes at the base of the tubercles and their subsequent differentiation in upper layers. Together, these studies identify the zebrafish breeding tubercles as specific epidermal structures sharing crucial features with the cornified mammalian epidermis. In addition, they unravel essential roles of TAp63 and p53 to promote both keratinocyte proliferation and their terminal differentiation by promoting Notch signalling and caspase 3 activity, ensuring formation and proper homeostasis of this self-renewing stratified epithelium.


Do large volume spacer devices reduce the systemic effects of high dose inhaled corticosteroids?
When used in high doses, inhaled corticosteroids may cause suppression of the hypothalamo-pituitary-adrenal axis. The influence of the mode of drug inhalation on the degree of this suppression is not clear. Hypothalamo-pituitary-adrenal function was assessed by measurement of 0900 h serum cortisol concentrations, a short tetracosactrin test, and 24 hour urine free cortisol excretion in 48 adults with asthma taking 1500-2500 micrograms beclomethasone dipropionate daily via a metered dose aerosol. Twelve patients had hypothalamo-pituitary-adrenal suppression, as judged by subnormal results from at least two of the three tests or (in one patient) by an abnormal insulin stress test response. These patients then changed to inhaling the same dose of beclomethasone dipropionate through a 750 ml spacer device (Volumatic). The endocrine tests were repeated from nine days to eight weeks later in 10 patients. Comparison with initial values showed that adding the spacing device caused an increase in the median 0900 h cortisol concentration from 126 nmol/l to 398 nmol/l, in the post-tetracosactrin cortisol concentration from 402 nmol/l to 613 nmol/l and in 24 hour urine free cortisol excretion from 54 nmol to 84 nmol. The rise in serum cortisol concentration in response to tetracosactrin did not change. Evidence of persisting hypothalamo-pituitary-adrenal axis suppression was present in only four of the 10 patients; the most pronounced improvements in function tended to occur in those who had never required long term oral corticosteroids. The results from this uncontrolled study suggest that asthmatic patients taking high dose beclomethasone dipropionate may minimise adverse effects by using a large volume spacer device.


Healthcare experiences of lesbian and bisexual women in Cape Town, South Africa.
Understanding the health needs and experiences of South African lesbian and bisexual women is imperative for implementing effective and inclusive public health strategies. Such understanding, however, is limited due to the exclusion of these women from most existing research on healthcare access in the region. This paper bridges that gap by investigating the healthcare experiences of lesbian and bisexual women in Cape Town. Data were gathered from 22 interviews with self-identified lesbian and bisexual community members and university students in the Cape Town area. Interviews explored obstacles women face in accessing affirming services, different experiences with public and private healthcare, fear of stigma/discrimination, availability of relevant sexual health information and suggestions to improve existing programmes. Findings suggest that South African lesbians and bisexual women may have a range of both positive and negative experiences in public and private health services, that they use protective strategies when 'coming out' and that they find that sexual health information pertinent to them is largely unavailable. These discussions contribute to a more inclusive understanding of the experiences of lesbian and bisexual women accessing healthcare and other services and help to inform providers, thereby enabling them to deliver more meaningful care to lesbian, gay, bisexual and transgender persons in South Africa.


Psychological impact of 131I radioprotection measures on thyroid cancer patients.
It is very well know nowadays that despite all the good and qualitative information available, the patients who are supposed to be treated with radioiodine for differentiated thyroid cancer suffer from a lot of concerns prior the treatment. The aim of our study is to investigate the level of anxiety and lessening of the concerns pre and post RIT (radioiodine therapy) using a dedicated, special designed questionnaire. A cross-sectional study of 54 differentiated thyroid cancer patients was conducted. Patients who met the inclusion criteria were invited to complete the radioprotection questionnaire pre and post RIT. The questionnaire comprises two sections (pre and post radioiodine treatment) with specific questions regarding aspects of radioprotection measures and the impact on the overall well-being. For uniform distribution of variables we used Pearson correlation and for monotonic relationship between variables, Spearman correlation. The pre-treatment questionnaire reported a strong confidence of the patients in the medical team, good and accurate information regarding the treatment, >50 % suffering from anxiety and concerns before the radioiodine treatment. The post treatment questionnaire revealed no fear of isolation, a lot of useful information and most of the patients would undergo another treatment, if necessary and also recommend it to others. The milestone in having a good and compliant patient remains a very good communication between the medical team and the patient. We are able to influence and change things and have fewer patients with fear from radioactivity and treatment concerns if we dedicate enough time to give them the adequate information in the best way so it will be correctly received.


Reactivity of anti-peptide and anti-poliovirus type 3 monoclonal antibodies with synthetic peptides.
Monoclonal antibodies were prepared from mice immunized with an 18-residue synthetic peptide with an amino acid sequence from a major antigenic sequence involved in the neutralization of type 3 poliovirus. Approximately 250 hybridomas secreted antibodies that reacted with the peptide but not the virus, two antibodies reacted with the virus but not the peptide and no antibody reacted with both. Conversely 26 monoclonal antibodies prepared from mice immunized with type 3 poliovirus and known to be directed against the appropriate sequence on the virus, generally failed to react with the peptide. These results might be expected if only a small proportion of the free or coupled peptide molecules adopt molecular conformations which resemble that of the homologous antigenic site in the virus. Antibodies specific for other antigens occasionally reacted well with the synthetic peptides, indicating that antibodies may bind to peptides of inappropriate sequence. The identification of antigenic sites by the use of synthetic peptides therefore requires considerable caution.


Plant litter loss exacerbates drought influences on grasslands.
Plant litter is known to affect soil, community, and ecosystem properties. However, we know little about the capacity of litter to modulate grassland responses to climate change. Using a 7-yr litter removal experiment in a semiarid grassland, here we examined how litter removal interacts with a 2-yr drought to affect soil environments, plant community composition, and ecosystem function. Litter loss exacerbates the negative impacts of drought on grasslands. Litter removal increased soil temperature but reduced soil moisture and nitrogen mineralization, which substantially increased the negative impacts of drought on primary productivity and the abundance of perennial rhizomatous graminoids. Moreover, complete litter removal shifted plant community composition from grass-dominated to forb-dominated and reduced species and functional group asynchrony, resulting in lower ecosystem temporal stability. Our results suggest that ecological processes that lead to reduction in litter, such as burning, grazing, and haying, may render ecosystems more vulnerable and impair the capacity of grasslands to withstand drought events.


Comparison of the intrusion effects on the maxillary incisors between implant anchorage and J-hook headgear.
Recently, miniscrews have been used to provide anchorage during orthodontic treatment, especially for incisor intrusion. Miniscrews during incisor intrusion are commonly used in implant orthodontics. Traditionally, effective incisor intrusion has been accomplished with J-hook headgear. In this study, we compared the effect of incisor intrusion, force vector, and amount of root resorption between implant orthodontics and J-hook headgear. Lateral cephalometric radiographs from 8 patients in the implant group and 10 patients in the J-hook headgear group were analyzed for incisor retraction. The estimated force vector was analyzed in the horizontal and vertical directions in both groups. Root resorption was also measured on periapical radiographs. In the implant group, significant reductions in overjet, overbite, maxillary incisor to palatal plane, and maxillary incisor to upper lip were observed after intrusion of the incisors. In the J-hook headgear group, significant reductions in overjet, overbite, maxillary incisor to upper lip, and maxillary incisor to SN plane were observed after intrusion of the incisors. There were significantly greater reductions in overbite, maxillary incisor to palatal plane, and maxillary incisor to upper lip in the implant group than in the J-hook headgear group. Estimated force analysis resulted in significantly more force in the vertical direction and less in the horizontal direction in the implant group. Furthermore, significantly less root resorption was observed in the implant group compared with the J-hook headgear group. The maxillary incisors were effectively intruded by using miniscrews as orthodontic anchorage without patient cooperation. The amount of root resorption was not affected by activating the ligature wire from the miniscrew during incisor intrusion.


CT features of intrathoracic gossypiboma (textiloma).
Here, we present chest radiography and computed tomography (CT) findings for three cases of mediastinal and pleural gossypiboma. Radiological manifestations varied according to the locations and chronicities of the gauze sponges and the types of reactions that they caused. CT analysis readily diagnosed gossypibomas in the early postoperative period by showing well-defined mediastinal- or pleural-based masses with hyperdense rims and central air bubbles. However, one patient presented in the late postoperative period, and a CT scan revealed a well-defined, solid pleural mass that was devoid of air bubbles; a correct diagnosis could not be established and the patient had to be reoperated on. Radiologists should be aware of different manifestations of this rare condition in cases of prior thoracic surgery and persistent respiratory symptoms.


Solitary Fibrous Tumor of Breast and Axilla: Clinicopathological Profile of Five Tumors With Comparison of Risk Stratification Models.
Introduction: Solitary fibrous tumor (SFT) is a fibroblastic tumor with malignant potential that is underpinned by a recurrent inv12(q13q13)-derived NAB2::STAT6 fusion. Breast and axilla are uncommon locations for this entity. Methods: Records of two academic institutions were electronically searched for breast and axillary SFTs. Clinical and pathologic data were reviewed. Literature review for breast or axillary SFTs was performed. Present study and previously reported tumors were stratified using five SFT risk models: original and modified Demicco metastatic risk, Salas local recurrence risk, Salas metastatic risk, and Thompson local recurrence risk. Results: Five patients with breast or axillary SFT were identified. Median age was 49 years, and median follow-up (available for four patients) was 82 months. Three patients showed no evidence of disease, and one developed recurrence. Literature review identified 58 patients with breast or axillary SFT. Median age was 54 years, and median follow-up (available for 35 patients) was 24 months. Thirty-one patients showed no evidence of disease, three developed recurrence, and one developed metastasis. Original and modified Demicco models and Thompson model showed the highest sensitivity; original and modified Demicco models and Salas metastatic risk model demonstrated the highest specificity. Kaplan-Meier models were used to assess recurrence-free probability (RFP). Original and modified Demicco models predicted RFP when stratified by "low risk" and "moderate/intermediate and high risk" tumor, though sample size was small. Conclusions: While many SFTs of breast and axilla remain indolent, a subset may develop recurrence and rarely metastasize. The modified Demicco risk model demonstrated optimal performance characteristics.


Subthalamic deep brain stimulation improves time perception in Parkinson's disease.
Alterations in temporal estimation have been observed in Parkinson's disease (PD) and have been associated with dopaminergic dysfunction. To investigate whether deep brain stimulation might reverse these abnormalities in PD, patients treated with electrode implantation for subthalamic deep brain stimulation were required to reproduce time intervals in different experimental conditions (off deep brain stimulation/off therapy, on deep brain stimulation/off therapy, on therapy/off deep brain stimulation). Patients treated with deep brain stimulation in off deep brain stimulation/off therapy displayed the anomalous pattern of responses typically observed in PD. When subthalamic deep brain stimulation was turned on these abnormalities were significantly attenuated. Our findings reveal that subthalamic deep brain stimulation improves time perception in PD patients, supporting the critical role of basal ganglia in this cognitive function, probably mediated by facilitated thalamo-cortical projections to the prefrontal cortex.


[Bronchiolitis obliterans with organizing pneumonia secondary to chemotherapy in a child with primary pericardial sarcoma].
The uncommon bronchiolitis obliterans organizing pneumonia can be idiopathic or caused by infection or medications. We report a 5 year old boy with pericardial sarcoma that was treated with chemotherapy (vincristine, doxorubicin, etoposide, cyclophosphamide) and radiotherapy. During the third cycle of chemotherapy, he developed progressive cough and dyspnea, needing oxygen therapy. Chest X-ray examination showed bilateral infiltrates. After discarding infectious etiologies, an open lung biopsy was performed, and the pathological examination showed a bronchiolitis obliterans organizing pneumonia. He was treated with prednisolone for 7 days, followed by prednisone for additional 45 days. He continued with fluticasone propionate as maintenance and remains in good condition with a chest X-ray showing mild interstitial images, after seven months of follow up.


Usefulness of 3'- 5' IS6110-RFLP genotyping and spoligotyping of Mycobacterium tuberculosis isolated in a tertiary hospital: a retrospective study detecting unsuspected epidemiological events.
A drug resistance survey involving Mycobacterium tuberculosis isolated from patients of a tertiary Hospital in the Rio de Janeiro city (RJ), Brazil, between the years 1996 and 1998 revealed a high frequency of isoniazid (HR) resistance. These isolates were revisited and genotyped. Patients came from different RJ neighborhoods and municipalities, and 70% were outpatients. Applying the 3' and 5' IS 6110 -RFLP and the Spoligotype genotyping methods, the clonal structure of this population was investigated obtaining a snapshot of past epidemiological events. The 3' clusters were subsequently 5' IS 6110 -RFLP typed. Spoligotyping was analyzed in the SITVIT2 database. Epidemiological relationships were investigated. The major lineage was T (54.4%), and SIT 53/T1 and SIT 535/T1 were the most frequent. The T1 sublineage comprises 12.8% of resistant strains and SIT 535 were assigned for 31.8% of them. Orphan patterns corresponded to 12% and 73.3% and belonged to the T lineage. One pattern was unlisted in the SITVIT2. The 5' IS 6110 -RFLP did not confirm 3/12 of the 3' IS 6110 -RFLP clusters. A combination of all methods decreased the number of clusters to three. Nosocomial transmission was associated with one cluster involving a hospital cupbearer. This event was suspected in a multidrug resistant-TB inpatient caregiver who harbored a mixed infection. The 3' IS 6110 clusters were associated with HR (p=0.046). These genotypic retrospective data may reflect a fraction of more extensive recent transmission in different communities that may be corroborated by the concentration of HR patients, and may serve as a database for further evolutionary and characterization evaluation of circulating strains and together with epidemiological data favors a more effective transmission control.


Compensatory activation of substance P biosynthesis by L-dihydroxyphenylalanine in striatonigral neurons of neonatal dopaminergic denervated rats.
Previous studies have shown that L-dihydroxyphenylalanine (L-DOPA) administration to adult rats that had been subjected to dopamingergic denervation with 6-hydroxydopamine during early postnatal period, led to a marked decrease in the levels of striatonigral substance P (SP). The present study examined the hypothesis of whether there is a compensatory activation of SP biosynthesis in order to replenish the L-DOPA-induced SP depletion. Three-day old Sprague-Dawley rat pups were lesioned with 6-hydroxydopamine and were challenged with L-DOPA at about 60 days of age. The animals were sacrificed 75 min, 6 hr or 24 hr after the L-DOPA administration. The levels of SP (in the striatum and substantia nigra) were determined by radioimmunoassay. The abundance of SP-encoding preprotachykinin (PPT) messenger RNA (mRNA) in the striatum was determined by Northern blot analysis and the abundance of individual PPT-mRNAs (alpha, beta and gamma) was determined by nuclease protection assays. Concentrations of dopamine and its acid metabolite, dihydroxyphenyl acetic acid were determined by high-pressure liquid chromatography with electrochemical detection. At the 75 min time point, L-DOPA produced a greater decrease in SP levels in the striatum and the substantia nigra, than that observed with lesion alone. The SP levels recovered to lesioned-control levels by 6 hr and remained at this level at the 24-hr time point. This recovery was accompanied by a marked increase in striatal SP-encoding PPT-mRNA abundance at 6 hr; mRNA levels were below lesioned-control value at 24 hr. There were no differential changes in the individual SP-encoding PPT-mRNAs.(ABSTRACT TRUNCATED AT 250 WORDS)


Crowding Effects on Energy-Transfer Efficiencies of Hetero-FRET Probes As Measured Using Time-Resolved Fluorescence Anisotropy.
Macromolecular crowding is prevalent in all living cells due to the presence of large biomolecules and organelles. Cellular crowding is heterogeneous and is known to influence biomolecular transport, biochemical reactions, and protein folding. Emerging evidence suggests that some cell pathologies may be correlated with compartmentalized crowding. As a result, there is a need for robust biosensors that are sensitive to crowding as well as quantitative, noninvasive fluorescence methods that are compatible with living cells studies. Here, we have developed a model that describes the rotational dynamics of hetero-Förster resonance energy transfer (FRET) biosensors as a means to determine the energy-transfer efficiency and donor-acceptor distance. The model was tested on wavelength-dependent time-resolved fluorescence anisotropy of hetero-FRET probes (mCerulean3-linker-mCitrine) with variable linkers in both crowded (Ficoll-70) and viscous (glycerol) solutions at room temperature. Our results indicate that the energy-transfer efficiencies of these FRET probes increase as the linker becomes shorter and more flexible in pure buffer at room temperature. In addition, the FRET probes favor compact structures with enhanced energy-transfer efficiencies and a shorter donor-acceptor distance in the heterogeneous, polymer-crowded environment due to steric hindrance. In contrast, the extended conformation of these FRET probes is more favorable in viscous, homogeneous environments with a reduced energy-transfer efficiency compared to those in pure buffer, which we attribute to reduced structural fluctuations of the mCerulean3-mCitrine FRET pair in the glycerol-enriched buffer. Our results represent an important step toward the application of quantitative and noninvasive time-resolved fluorescence anisotropy of hetero-FRET probes to investigate compartmentalized macromolecular crowding and protein-protein interactions in living cells as well as in controlled environments.


[Effect of different doses of ethanol on the release of enzymes and lipids from the perfused rat liver].
The perfusion in situ of the rat liver reveals appearance of enzymes of different subcellular localization, potassium ions and lipid components in the perfusate. Ethanol introduction at different doses induces the most expressed changes in the activity of catalase, lactate dehydrogenase, arginase and alanine aminotransferase and in the potassium removal from hepatocytes. A degree of the evoked changes depends on the dose.


Research Progress on the Mechanism of the Acupuncture Regulating Neuro-Endocrine-Immune Network System.
As one of the conventional treatment methods, acupuncture is an indispensable component of Traditional Chinese Medicine. Currently, acupuncture has been partly accepted throughout the world, but the mechanism of acupuncture is still unclear. Since the theory of the neuro-endocrine-immune network was put forward, new insights have been brought into the understanding of the mechanism of acupuncture. Studies have proven that acupuncture is a mechanical stimulus that can activate local cell functions and neuroreceptors. It also regulates the release of related biomolecules (peptide hormones, lipid hormones, neuromodulators and neurotransmitters, and other small and large biomolecules) in the microenvironment, where they can affect each other and further activate the neuroendocrine-immune network to achieve holistic regulation. Recently, growing efforts have been made in the research on the mechanism of acupuncture. Some researchers have transitioned from studying the mechanism of acupuncture as a single linear pathway to using systems approaches, including metabolomics, genomics, proteomics and biological pathway analysis. This review summarizes the research progress on the neuro-endocrine-immune network related mechanism of acupuncture and discusses its current challenges and future directions.


Prairie strips improve biodiversity and the delivery of multiple ecosystem services from corn-soybean croplands.
Loss of biodiversity and degradation of ecosystem services from agricultural lands remain important challenges in the United States despite decades of spending on natural resource management. To date, conservation investment has emphasized engineering practices or vegetative strategies centered on monocultural plantings of nonnative plants, largely excluding native species from cropland. In a catchment-scale experiment, we quantified the multiple effects of integrating strips of native prairie species amid corn and soybean crops, with prairie strips arranged to arrest run-off on slopes. Replacing 10% of cropland with prairie strips increased biodiversity and ecosystem services with minimal impacts on crop production. Compared with catchments containing only crops, integrating prairie strips into cropland led to greater catchment-level insect taxa richness (2.6-fold), pollinator abundance (3.5-fold), native bird species richness (2.1-fold), and abundance of bird species of greatest conservation need (2.1-fold). Use of prairie strips also reduced total water runoff from catchments by 37%, resulting in retention of 20 times more soil and 4.3 times more phosphorus. Corn and soybean yields for catchments with prairie strips decreased only by the amount of the area taken out of crop production. Social survey results indicated demand among both farming and nonfarming populations for the environmental outcomes produced by prairie strips. If federal and state policies were aligned to promote prairie strips, the practice would be applicable to 3.9 million ha of cropland in Iowa alone.


Quality of the diagnostic process in patients presenting with symptoms suggestive of bladder or kidney cancer: a systematic review.
In urological cancers, sex disparity exists for survival, with women doing worse than men. Suboptimal evaluation of presenting symptoms may contribute. We performed a systematic review examining factors affecting the quality of the diagnostic process of patients presenting with symptoms of bladder or kidney cancer. We searched Medline, Embase and the Cochrane Library from 1 January 2000 to 13 June 2019. We focused on one of the six domains of quality of healthcare: timeliness, and examined the quality of the diagnostic process more broadly, by assessing whether guideline-concordant history, examination, tests and referrals were performed. Studies describing the factors that affect the timeliness or quality of the assessment of urinary tract infections, haematuria and lower urinary tract symptoms in the context of bladder or kidney cancer, were included. Data extraction and quality assessment were independently performed by two authors. Due to the heterogeneity of study design and outcomes, the results could not be pooled. A narrative synthesis was performed. 28 studies met review criteria, representing 583 636 people from 9 high-income countries. Studies were based in primary care (n=8), specialty care (n=12), or both (n=8). Up to two-thirds of patients with haematuria received no further evaluation in the 6 months after their initial visit. Urinary tract infections, nephrolithiasis and benign prostatic conditions before cancer diagnosis were associated with diagnostic delay. Women were more likely to experience diagnostic delay than men. Patients who first saw a urologist were less likely to experience delayed evaluation and cancer diagnosis. Women, and patients with non-cancerous urological diagnoses just prior to their cancer diagnosis, were more likely to experience lower quality diagnostic processes. Risk prediction tools, and improving guideline ambiguity, may improve outcomes and reduce sex disparity in survival for these cancers.


Immunoblotting with monoclonal antibodies: importance of the blocking solution.
Four commonly used blocking agents, i.e., fetal calf serum, mammalian gelatin-Nonidet-P40, fish gelatin-Nonidet-P40, and defatted powdered milk were compared with respect to their efficiency to block the nonspecific background and to promote maximal immunoreactivity of monoclonal antibodies against human intestinal sucrase-isomaltase during immunoblotting. Two of five monoclonal antibodies were found to react with the electroblotted enzyme. However, one of the reacting antibodies gave optimal results with fish gelatin-Nonidet-P40 and the other with defatted powdered milk, while fetal calf serum lead to unacceptably high backgrounds. The results suggest that some of the difficulties encountered with monoclonal antibodies in immunoblotting may be due to inappropriate blocking conditions.


Vinculin is an essential component for normal myofibrillar arrangement in fetal mouse cardiac myocytes.
Vinculin is a cytoskeletal protein that is believed to be an essential component in the linkage of cytoskeletal actin filaments to the plasma membrane. To investigate the precise function of vinculin in the development of cardiac myofibrils, antisense oligodeoxynucleotides complementary to vinculin mRNA were used to perturb the expression of the protein during myofibril assembly and arrangement in mouse cardiac myocytes. Fetal (day 18-20 post-conception) mouse cardiac myocytes were isolated by collagenase digestion, separated by Percoll density gradient centrifugation, and plated on aligned collagen gels. By 72 h of culture, mouse myocytes displayed an elongated in vivo-like phenotype in parallel with the aligned fibrils of the collagen gels with polarized arrays of myofibrils. Two different antisense oligonucleotides (20-mer) altered the formation of the tissue-like phenotype of myocytes. These antisense oligonucleotides suppressed vinculin protein expression at 43.5+/-26.8% and 48.7+/-20.9% when compared to myocytes that were not treated. Examination of these myocytes by confocal scanning laser and transmission electron microscopy revealed a disruption of the aligned in vivo-like phenotype, assembly of thick and thin filaments, and formulation of Z-bands. Random sequence 20-mer oligonucleotides used as controls had little detectable effect on vinculin protein expression (94.2+/-14.8%), cell shape, normal alignment or assembly of myofibrils. These results indicate that vinculin is a critical cytoskeletal component, that functions in the determination of cell shape and the arrangement and organization of developing myofibrils.


Functional frameworks of illumination revealed by probe disk technique.
We used a novel probe disk technique to test for the existence of functional frames of reference for lightness perception in complex images. Thirteen identical gray disks were electronically pasted into the photograph Trastevere, which shows two large regions of sunlight and shadow. Observers matched the lightness of each disk with a Munsell scale. The data revealed a framework effect. That is, lightness differences within either the sunlight or shadow region were small relative to the pronounced step function at the framework boundary. Additional experiments testing the perceived embeddedness of the disks showed that the framework effect was increased when disk size and shape were altered to conform to the perspective shown in the photograph and when the disks were blurred slightly to conform to the graininess of the photograph. The effect was further increased when the photograph was viewed through a pinhole and when the disks were presented one by one. The effect was reduced when paper disks of equal luminance and visual angle were pasted onto the glass front of the CRT screen. When the sunlight framework was covered with black paper, the remained disks within the shadow region appeared white, as predicted by the anchoring theory (A. Gilchrist, 2006).


Zika Virus on YouTube: An Analysis of English-language Video Content by Source.
The purpose of this study was to describe the source, length, number of views, and content of the most widely viewed Zika virus (ZIKV)-related YouTube videos. We hypothesized that ZIKV-related videos uploaded by different sources contained different content. The 100 most viewed English ZIKV-related videos were manually coded and analyzed statistically. Among the 100 videos, there were 43 consumer-generated videos, 38 Internet-based news videos, 15 TV-based news videos, and 4 professional videos. Internet news sources captured over two-thirds of the total of 8 894 505 views. Compared with consumer-generated videos, Internet-based news videos were more likely to mention the impact of ZIKV on babies (odds ratio [OR], 6.25; 95% confidence interval [CI], 1.64 to 23.76), the number of cases in Latin America (OR, 5.63; 95% CI, 1.47 to 21.52); and ZIKV in Africa (OR, 2.56; 95% CI, 1.04 to 6.31). Compared with consumer-generated videos, TV-based news videos were more likely to express anxiety or fear of catching ZIKV (OR, 6.67; 95% CI, 1.36 to 32.70); to highlight fear of ZIKV among members of the public (OR, 7.45; 95% CI, 1.20 to 46.16); and to discuss avoiding pregnancy (OR, 3.88; 95% CI, 1.13 to 13.25). Public health agencies should establish a larger presence on YouTube to reach more people with evidence-based information about ZIKV.


[Diagnosis in forensic medical practice].
The author performed the analysis of the essence and patogenetic structure of diagnosis in forensic medical practice, the similarities and differences between forensic medical, pathological morphological, and clinical diagnoses, and their significance for the forensic medical expert conclusion.


Spectroscopic and Structural Study of Some Oligosilanylalkyne Complexes of Cobalt, Molybdenum and Nickel.
Metal induced stabilization of α-carbocations is well known for cobalt- and molybdenum complexed propargyl cations. The same principle also allows access to reactivity enhancement of metal coordinated halo- and hydrosilylalkynes. In a previous study, we have shown that coordination of oligosilanylalkynes to the dicobalthexacarbonyl fragment induces striking reactivity to the oligosilanyl part. The current paper extends this set of oligosilanylalkyne complexes to a number of new dicobalthexacarbonyl complexes but also to 1,2-bis(cyclopentadienyl)tetracarbonyldimolybdenum and (dippe)Ni complexes. NMR-Spectroscopic and crystallographic analysis of the obtained complexes clearly show that the dimetallic cobalt and molybdenum complexes cause rehybridization of the alkyne carbon atoms to sp³, while in the nickel complexes one π-bond of the alkyne is retained. For the dicobalt and dimolybdenum complexes, strongly deshielded 29Si NMR resonances of the attached silicon atoms indicate enhanced reactivity, whereas the 29Si NMR shifts of the respective nickel complexes are similar to that of respective vinylsilanes.


Preschool Interpersonal Relationships Predict Kindergarten Achievement: Mediated by Gains in Emotion Knowledge.
Using longitudinal data, this study tested a model in which preschool interpersonal relationships promoted kindergarten achievement in a pathway mediated by growth in emotion knowledge. The sample included 164 children attending Head Start (14% Hispanic-American, 30% African-American, 56% Caucasian; 56% girls). Preschool interpersonal relationships were indexed by student-teacher relationship closeness and positive peer interactions. Two measures of emotion knowledge (identifying emotions in photographs, recognizing emotions in stories) were assessed at the start and end of the preschool year. Structural equation models revealed that positive interpersonal relationships (with teachers and peers) predicted gains in emotion knowledge (identification, recognition) during the preschool year. Positive interpersonal relationships in preschool also predicted kindergarten achievement (controlling for initial preschool achievement); however, this association was mediated by gains in emotion knowledge during the preschool year. Implications are discussed for school readiness programs serving economically-disadvantaged children.


[The capillary blood flow in the lungs of patients with chronic laryngeal and tracheal stenosis].
Altogether 19 patients with chronic laryngo- and tracheostenosis of different etiology were investigated. All the patients were broken into 3 groups (1--less than 1 year, 2-1-3 years, 3--over 3 years) with regard to the time of canulla bearing. The patients were examined with 99mTc-human serum albumin particles. Major disorders in the capillary blood supply of both lungs were detected. These changes were focal-diffuse. Abnormalities were mainly observed in the apical area of both lungs and in the mediastinal zone. Redistribution of the pulmonary blood supply in the direction of the upper parts of the lungs was revealed. Lung abnormalities increased depending on the time of canulla bearing.


Patient-initiated versus fixed-interval patient-reported outcome-based follow-up in outpatients with epilepsy: a pragmatic randomized controlled trial.
The use of patient-reported outcome (PRO) could potentially contribute to the reorganization of the health care system. AmbuFlex is a PRO system used in remote patient monitoring, in which questionnaires are sent to patients at fixed intervals. The PRO data are used by clinicians to decide whether patients need clinical attention. Better self-management and cost-saving follow-up activities may be achieved by letting patients initiate need of contact. We evaluated the effects of patient-initiated PRO-based outpatient follow-up on health care resource utilization, quality of care, and the patient perspective. We conducted a parallel two-arm pragmatic randomized controlled trial at the Department of Neurology, Aarhus University Hospital, Denmark. Outpatients with epilepsy (≥ 15 years old), attending fixed-interval PRO-based follow-up with web-based questionnaires, were randomly assigned in a ratio of 0.55:0.45 to either 1) patient-initiated PRO-based follow-up (open access telePRO) or 2) fixed-interval PRO-based follow-up (standard telePRO). The primary outcome was the number of outpatient hospital contacts related to epilepsy retrieved from a regional registry. Hospitals admissions and emergency room visits were also assessed. Secondary self-reported outcomes including general health, well-being, health literacy, self-efficacy, number of seizures, side effects, confidence, safety, and satisfaction were retrieved from questionnaires. Data were analyzed by the intention-to-treat and per-protocol approaches. Between January 2016 and July 2016, 593 patients were randomized to either open access telePRO (n = 346) or standard telePRO (n = 247). At 18 months, no statistically significant differences were found between the arms regarding number of telephone consultations or outpatient visits. Patients in the open access arm had a slightly lower, statistically significant number of emergency room visits than patients in the standard arm. Self-reported mental well-being in the open access arm was slightly, statistically significantly lower than in the standard arm. Other secondary outcomes did not differ statistically significantly between arms. This study did not find, as hypothesized, less use of health care resources or improved patient self-management or satisfaction in the patient-initiated PRO-based initiative compared to fixed-interval PRO-based follow-up. Patient-initiated PRO-based follow-up may be used as an alternative to fixed-interval PRO-based follow-up in patients who prefer this approach, but there is insufficient evidence for recommending a system-wide shift to patient-initiated PRO-based follow-up. Registered 4 February 2016 with ClinicalTrials.gov: NCT02673580 .


Comparison of clinical findings and short-term survival between horses with intestinal entrapment in the gastrosplenic ligament and horses with intestinal entrapment in the epiploic foramen.
OBJECTIVE To compare clinical findings and short-term outcome for horses with intestinal entrapment in the gastrosplenic ligament (GLE) with those of horses with intestinal entrapment in the epiploic foramen (EFE). DESIGN Retrospective case-control study. ANIMALS 43 horses with GLE (cases) and 73 horses with EFE (controls). PROCEDURES Medical records of horses examined because of colic at a veterinary teaching hospital between 1992 and 2012 were reviewed. Signalment was extracted from medical records for all horses with colic (colic population), and additional information regarding colic history, clinical findings, treatments, and outcome was extracted from the records of horses in which GLE or EFE was diagnosed during surgery or necropsy. Signalment was compared between the colic population and the case and control populations. Clinical findings and short-term outcome were compared between the cases and controls. RESULTS The proportions of middle-aged horses and geldings in both the case and control groups were greater than those in the colic population. Mean heart rate and blood and peritoneal fluid lactate concentrations in horses with EFE were significantly greater than those for horses with GLE. The proportion of horses that underwent surgery and were discharged from the hospital (short-term survival rate) did not differ between the GLE (22/25 [88%]) and EFE (29/34 [85%]) groups. CONCLUSIONS AND CLINICAL RELEVANCE Compared with the colic population, results suggested middle-aged geldings might be predisposed to GLE and EFE. The short-term survival rate was similar between the GLE and EFE groups even though horses with EFE had more severe systemic derangements than did horses with GLE.


mRNAs and Protein Synthetic Machinery Localize into Regenerating Spinal Cord Axons When They Are Provided a Substrate That Supports Growth.
Although intra-axonal protein synthesis is well recognized in cultured neurons and during development in vivo, there have been few reports of mRNA localization and/or intra-axonal translation in mature CNS axons. Indeed, previous work indicated that mature CNS axons contain much lower quantities of translational machinery than PNS axons, leading to the conclusion that the capacity for intra-axonal protein synthesis is linked to the intrinsic capacity of a neuron for regeneration, with mature CNS neurons showing much less growth after injury than PNS neurons. However, when regeneration by CNS axons is facilitated, it is not known whether the intra-axonal content of translational machinery changes or whether mRNAs localize into these axons. Here, we have used a peripheral nerve segment grafted into the transected spinal cord of adult rats as a supportive environment for regeneration by ascending spinal axons. By quantitative fluorescent in situ hybridization combined with immunofluorescence to unambiguously distinguish intra-axonal mRNAs, we show that regenerating spinal cord axons contain β-actin, GAP-43, Neuritin, Reg3a, Hamp, and Importin β1 mRNAs. These axons also contain 5S rRNA, phosphorylated S6 ribosomal protein, eIF2α translation factor, and 4EBP1 translation factor inhibitory protein. Different levels of these mRNAs in CNS axons from regenerating PNS axons may relate to differences in the growth capacity of these neurons, although the presence of mRNA transport and likely local translation in both CNS and PNS neurons suggests an active role in the regenerative process. Although peripheral nerve axons retain the capacity to locally synthesize proteins into adulthood, previous studies have argued that mature brain and spinal cord axons cannot synthesize proteins. Protein synthesis in peripheral nerve axons is increased during regeneration, and intra-axonally synthesized proteins have been shown to contribute to nerve regeneration. Here, we show that mRNAs and translational machinery are transported into axons regenerating from the spinal cord into the permissive environment of a peripheral nerve graft. Our data raise the possibility that spinal cord axons may make use of localized protein synthesis for regeneration.


Detection of Echinococcus multilocularis and other foodborne parasites in fox, cat and dog faeces collected in kitchen gardens in a highly endemic area for alveolar echinococcosis.
Echinococcus multilocularis, Toxoplasma gondii and Toxocara spp. are foodborne parasites whose eggs or oocysts are spread in the environment via canid or felid faeces. They can cause infections in humans following the raw consumption of contaminated fruit or vegetables. In this study, their occurrence was investigated by quantitative polymerase chain reaction (qPCR) in 254 carnivore faeces deposited in 94 kitchen gardens of northeastern France that were sampled between two and six times from October 2011 to April 2013. Less than 25% of the sampled kitchen gardens contained more than 75% of the collected faeces. Of the 219 faeces that could be attributed to an emitter, cat accounted for 58%, fox for 32% and dog for 10%. Echinococcus multilocularis was detected in 35%, 11% and 7% of fox, dog and cat faeces, respectively, and Toxocara spp. in 33%, 12% and 5.5% of cat, fox and dog faeces, respectively. Toxoplasma gondii was detected in 2/125 cat faeces and 2/21 dog faeces. The 34 faeces that tested positive for E. multilocularis were found in only 19 out of the 94 sampled kitchen gardens, and the 40 faeces that tested positive for Toxocara spp. were found in 28 of them. Consequently, some kitchen gardens appeared particularly at risk of human exposure to foodborne parasites, including E. multilocularis responsible for alveolar echinococcosis (AE), which is a serious zoonosis. In endemic areas, kitchen garden owners should be informed about the zoonotic risk linked to carnivore faeces deposits and encouraged to set up preventive measures.


[Cytochemical demonstration of an endogenous inhibitor of Na-K ATPase and its relationship to familial arterial hypertension].
Acute volume expansion, an increase in sodium intake and a restraint on sodium excretion endow the plasma with the capacity to cause a natriuresis, to inhibit sodium transport and to stimulate vascular reactivity. One natriuretic substance, the atrial natriuretic peptide, has been identified. Cytochemical techniques can detect the presence of a Na-K ATPase inhibitor in the plasma of normal man and the rat, the concentration of which is controlled by salt intake. The substance responsible appears to originate in the hypothalamus. The plasma concentration of the cytochemically detectable Na-K ATPase inhibitor is substantially raised in the plasma of patients with essential hypertension, of the spontaneously hypertensive rat and of the Milan hypertensive rat. An hypothesis is put forward that links salt intake, a genetic renal lesion, the endogenous Na-K ATPase inhibitor, the atrial natriuretic peptide, and the substance responsible for vascular reactivity, with the rise in arterial pressure in hereditary forms of hypertension.


Real-world safety and effectiveness of dapagliflozin in people living with type 1 diabetes in Spain: The Dapa-ON multicenter retrospective study.
To assess real-world safety and effectiveness of dapagliflozin in people living with type 1 diabetes mellitus (T1DM). We conducted a multicenter retrospective study in Spain including data from 250 people living with T1DM receiving dapagliflozin as add-on therapy to insulin (80.8% on-label use). The number of diabetic ketoacidosis (DKA) events was calculated over a 12-month follow-up (primary outcome). Changes in body weight, HbA1c, total daily insulin dose, and continuous glucose monitoring (CGM) metrics from baseline (at dapagliflozin prescription) to 12 months were also evaluated. A total of five DKA events (2.4% [95% CI 0.3;4.5] were reported in patients with a 12-month follow-up, n= 207): two events related to insulin pump malfunction, two events related to concomitant illnesses, and one event related to insulin dose omission. DKA events were more frequent among insulin pump users (7.7% versus 1.2%). Four of the reported DKA events occurred within the first six months after initiation of dapagliflozin. No deaths or persistent sequelae due to DKA were reported. No severe hypoglycemia episodes were reported. Significant reductions in mean body weight (-3.3 kg), HbA1c (-0.6%), and total daily insulin dose (- 8.6%), P < 0.001, were observed 12 months after dapagliflozin prescription. Significant improvements in TIR (+ 9.3%), TAR (-7.2%), TBR (-2.5%), and coefficient of variation (- 5.1%), P < 0.001, were also observed in the subgroup of patients with available CGM data. Finally, an improvement in urinary albumin-to-creatinine ratio (UACR) was found among participants with UACR ≥ 30 mg/g at baseline (median decrease of 99 mg/g in UACR, P = 0.001). The use of dapagliflozin in people living with T1DM has an appropriate safety profile after careful selection of participants and implementation of strategies to reduce the risk of DKA (i.e., prescribed according to the recommendations of the European Medicines Agency), and also leads to clinical improvements in this population.


Porosity control in mesoporous polymers using CO2-swollen block copolymer micelles as templates and their use as catalyst supports.
Mesoporous polymers with tunable large mesopores and thin mesopore walls were synthesized through a CO2-swollen micelle templating route. The mesopore size and porosity properties of the polymers can be easily modulated by adjusting CO2 pressure. The as-synthesized mesocellular polymers are excellent candidate supports for preparing heterogeneous catalysts.


[Intravenous gamma globulin in newborns].
Eight newborns with a mean gestational age of 36.1 weeks and a mean weight of 2863 grams were treated intravenously with 3% gammaglobulin (Sandoglobulina). We used two different infusion rates: one, of 0.5-1.5 mg/kg/min., and a second one, of 7-9 mg/kg/min. Close cardiopulmonary monitoring was carried out by bioelectronic means. The changes of heart rate, mean blood pressure and respiratory rate before and during the gammaglobulin infusion were recorded. We did not find any significant variation between the values recorded before and during the infusion. These values were all within tolerable physiological ranges except for those for respiratory rate, where 7 out of 8 patients had mild tachypnea. Stable sick newborn because of bacterial infections tolerated well the infusion of intravenous gammaglobulin at rates of 0.5-9.0 mg/kg/min., when the doses used were between 0.25 mg/kg/day and 1.0 gm/kg/day, for 4-6 days.


PML/RARA inhibits expression of HSP90 and its target AKT.
Essential for cell survival, the 90 kD Heat Shock Proteins (HSP90) are molecular chaperons required for conformational stabilization and trafficking of numerous client proteins. Functional HSP90 is required for the stability of AKT, a serine-threonine kinase phosphorylated in response to growth factor stimulation. AKT plays a crucial regulatory role in differentiation, cell cycle, transcription, translation, metabolism and apoptosis. Acute promyelocytic leukaemia (APL) is characterized by the presence of the promyelocytic leukaemia/retinoic acid receptor alpha (PML/RARA) fusion protein, which deregulates expression of several genes involved in differentiation and apoptosis. Here, we report inhibition of HSP90AA1 and HSP90AB1 isomer transcription in blasts isolated from patients with APL, associated with reduction of HSP90 protein expression and loss of control on AKT protein phosphorylation. We show that in vitro treatment of PML/RARA expressing cells with all-trans retinoic acid (ATRA) up-regulates HSP90 expression and stabilizes AKT. Addition of the HSP90-inhibitor 17-(allylamino)-17-demethoxygeldanamycin in combination with ATRA, blocks upregulation of AKT protein, indicating that HSP90 is necessary for ATRA action on AKT. This is the first report proving that expression of HSP90 isomers are directly and differentially repressed by PML/RARA, with critical results on cellular homeostasis of target proteins, such as AKT, in APL blasts.


Massively parallel sequencing and identification of genes for primary lymphoedema: a perfect fit.
Primary lymphoedema is a clinically and genetically heterogeneous group of disorders characterized by disruption of the lymphatic system. To date, the majority of the causative genes in primary lymphoedema have been identified through linkage analysis in large families with multiple affected subjects. Studies aimed at isolating additional genes responsible for primary lymphoedema have been hampered by cohorts comprised primarily of sporadic cases and small affected kindreds. In the absence of genetic heterogeneity, recent development of massively parallel DNA sequencing technology, specifically exome sequencing, has provided novel paradigms for disease gene identification in such cohorts. In this review, we summarize the novel approaches to disease gene discovery with massively parallel sequencing also known as Next Generation Sequencing (NGS), and show how the selection of unrelated affected cases from clinically homogenous phenotypic subclassifications is proving to be a successful approach for disease gene discovery in primary lymphoedema.


A biochemical study on the kinetics of kininogen in asphyxiated newborn.
From the viewpoint of fetal distress related disseminated intravascular coagulation (DIC), the Hageman Factor, kinin-kallikrein system were investigated and on the other hand the adaptation of the normal newborn to extrauterine life was also evaluated. (1) The Kininogen volume of the cord venous blood was approximately 1/2-1/3 or there abouts and was remarkable in serious cases of asphyxiated newborn. (2) In a similar manner the Hageman factor decreases in asphyxiated newborn. On the other hand, SFMC increases in aspyxiation as well as FDP. According to these results, there seems to be a coagulation obstruction due to consumption. Or it may be interpreted as an inhibition of kinin separation. In any event it may be considered as a rational contribution to the living body. (3) Based on the above, in cases of serious asphyxiation, DIC may possibly play a role and in these cases and the kininkallikrein system may be one of the mediators.


Do strict formularies replicate failure for patients with schizophrenia?
We measure the impact of Medicaid formulary restrictions (FRs) on the rate at which patients who previously failed a drug therapy for schizophrenia are returned to that therapy. We collect drug-level information on FRs in state Medicaid programs and examine claims of noninstitutionalized Medicaid enrollees with schizophrenia. A difference-in-differences technique is used to compute the change in the probability of adverse outcomes before and after a state adopts an FR. This change is compared with the change in failure probabilities in states with no FRs. Regardless of FRs, patients tend to resume the same drug after an adverse medical event. In 2005, 69% of inpatient mental health-related admissions resulted in patients resuming the same therapy within 6 months of the event, and 63% of patients resumed the same drug after a mental health-related emergency department admission. In states where FRs limit access to all atypicals, the likelihood of a patient resuming the same atypical after having ceased treatment for at least 30 days increases by 20.1% relative to patients in states without restrictions. Additionally, patients in states that impose FRs on all atypicals are 11.6% more likely to discontinue all treatments. FR may increase the likelihood that patients will return to failed treatments or cease treatment altogether. Although formularies are designed to reduce drug spending, an unintended consequence may be an increase in the use of other services needed to treat patients with schizophrenia.


Tracking technology: lessons learned in two health care sites.
The aim of this study is to describe the process of staff and patient adoption and compliance of a real-time locating system (RTLS) across two health care settings and present lessons learned. While previous work has examined the technological feasibility of tracking staff and patients in a health care setting in real-time, these studies have not described the critical adoption issues that must be overcome for deployment. The ability to track and monitor individual staff and patients presents new opportunities for improving workflow, patient health and reducing health care costs. A RTLS is introduced in both a long-term care and a polytrauma transitional rehabilitation program (PTRP) in a Veterans Hospital to track staff and patient locations and five lessons learned are presented from our experiences and responses to emergent technological, work-related and social barriers to adoption. We conclude that successful tracking in a health care environment requires time and careful consideration of existing work, policies and stakeholder needs which directly impact the efficacy of the technology.


Parathyroid hormone-related protein mRNA in sheep endometrium and myometrium during late gestation and labor.
To determine quantitative changes of parathyroid hormone-related protein (PTHrP) mRNA in sheep myometrium and endometrium during late gestation and labor. Tissues were obtained from 22 pregnant ewes under halothane anesthesia: early controls at 131 days' gestational age (dGA) not in labor (ECNL; n = 6); during cortisol-induced premature labor (CPL at 131 dGA; n = 6); in term spontaneous labor (STL at 140-145 dGA; n = 5); or term control animals not in labor (TCNL at 140-145 dGA; n = 5). Total RNA was extracted and subjected to Northern blot analysis. Blots were probed with a human PTHrP cDNA probe, stripped, and rehybridized with an 18s rRNA nucleotide probe to normalize PTHrP mRNA levels. Endometrial PTHrP mRNA:18s was unaffected by gestational age or labor in tissues from the four groups. In contrast, myometrial PTHrP mRNA:18s ratio was decreased when TCNL and STL were compared with both ECNL and CPL groups (P < .05). Significant changes occur at the end of gestation in pregnant ovine myometrium PTHrP mRNA but not in the endometrium. In myometrium PTHrP mRNA levels were down-regulated at term regardless of whether labor was present. It seems that PTHrP mRNA levels in myometrium are related to gestational age rather than to labor per se in sheep. We hypothesize that PTHrP may play a role in maintaining uterine quiescence until term, at which time levels fall allowing myometrial contractile activity to increase unopposed by PTHrP.


The Glutamate Receptor-Like Protein GLR3.7 Interacts With 14-3-3ω and Participates in Salt Stress Response in Arabidopsis thaliana.
Ionotropic glutamate receptors (iGluRs) are ligand-gated cation channels that mediate fast excitatory neurotransmission in the mammalian central nervous system. In the model plant Arabidopsis thaliana, a family of 20 glutamate receptor-like proteins (GLRs) shares similarities to animal iGluRs in sequence and predicted secondary structure. However, the function of GLRs in plants is little known. In the present study, a serine site (Ser-860) of AtGLR3.7 phosphorylated by a calcium-dependent protein kinase (CDPK) was identified and confirmed by an in vitro kinase assay. Using a bimolecular fluorescence complementation and quartz crystal microbalance analyses, the physical interaction between AtGLR3.7 and the 14-3-3ω protein was confirmed. The mutation of Ser-860 to alanine abolished this interaction, indicating that Ser-860 is the 14-3-3ω binding site of AtGLR3.7. Compared with wild type, seed germination of the glr3.7-2 mutant was more sensitive to salt stress. However, the primary root growth of GLR3.7-S860A overexpression lines was less sensitive to salt stress than that of the wild-type line. In addition, the increase of cytosolic calcium ion concentration by salt stress was significantly lower in the glr3.7-2 mutant line than in the wild-type line. Moreover, association of 14-3-3 proteins to microsomal fractions was less in GLR3.7-S860A overexpression lines than in GLR3.7 overexpression line under 150 mM NaCl salt stress condition. Overall, our results indicated that GLR3.7 is involved in salt stress response in A. thaliana by affecting calcium signaling.


Knee Kinematics of Healthy Adults Measured Using Biplane Radiography.
A dataset of knee kinematics in healthy, uninjured adults is needed to serve as a reference for comparison when evaluating the effects of injury, surgery, rehabilitation, and age. Most currently available datasets that characterize healthy knee kinematics were developed using conventional motion analysis, known to suffer from skin motion artifact. More accurate kinematics, obtained from bone pins or biplane radiography, have been reported for datasets ranging in size from 5 to 15 knees. The aim of this study was to characterize tibiofemoral kinematics and its variability in a larger sample of healthy adults. Thirty-nine knees were imaged using biplane radiography at 100 images/s during multiple trials of treadmill walking. Multiple gait trials were captured to measure stance and swing-phase knee kinematics. Six degrees-of-freedom kinematics were determined using a validated volumetric model-based tracking process. A bootstrapping technique was used to define average and 90% prediction bands for the kinematics. The average ROM during gait was 7.0 mm, 3.2 mm, and 2.9 mm in anterior/posterior (AP), medial/lateral (ML), and proximal/distal (PD) directions, and 67.3 deg, 11.5 deg, and 3.7 deg in flexion/extension (FE), internal/external (IE), and abduction/adduction (AbAd). Continuous kinematics demonstrated large interknee variability, with 90% prediction bands spanning approximately ±4 mm, ±10 mm, and ±5 mm for ML, AP, and PD translations and ±15 deg, ±10 deg, and ±6 deg in FE, IE, and AbAd. This dataset suggests substantial variability exists in healthy knee kinematics. This study provides a normative database for evaluating knee kinematics in patients who receive conservative or surgical treatment.


Antibiotic treatment durations for pyogenic liver abscesses: A systematic review.
We sought to systematically review the existing research on pyogenic liver abscesses to determine what data exist on antibiotic treatment durations. We conducted a systematic review and meta-analysis of contemporary medical literature from 2000 to 2020, searching for studies of pyogenic liver abscesses. The primary outcome of interest was mean antibiotic treatment duration, which we pooled by random-effects meta-analysis. Meta-regression was performed to examine characteristics influencing antibiotic durations. Sixteen studies (of 3,933 patients) provided sufficient data on antibiotic durations for pooling in meta-analysis. Mean antibiotic durations were highly variable across studies, from 8.4 (SD 5.3) to 68.9 (SD 30.3) days. The pooled mean treatment duration was 32.7 days (95% CI 24.9 to 40.6), but heterogeneity was very high (I2 = 100%). In meta-regression, there was a non-significant trend towards decreased mean antibiotic treatment durations over later study years (-1.14 days/study year [95% CI -2.74 to 0.45], p = 0.16). Mean treatment duration was not associated with mean age of participants, percentage of infections caused by Klebsiella spp, percentage of patients with abscesses over 5 cm in diameter, percentage of patients with multiple abscesses, and percentage of patients receiving medical management. No randomized trials have compared treatment durations for pyogenic liver abscess, and no observational studies have reported outcomes according to treatment duration. Among studies reporting on antibiotic durations for pyogenic liver abscess, treatment practices are highly variable. This variability does not seem to be explained by differences in patient, pathogen, abscess, or management characteristics. Future RCTs are needed to guide optimal treatment duration for patients with this complex infection.


Analysis of the n-GaN electrochemical etching process and its mechanism in oxalic acid.
We studied the wet electrochemical etching of n-GaN films in oxalic acid. The electrooxidation processes occur in a potentiostatic mode in the voltage range of 5 to 20 V. We described the formation of the porous n-GaN layer structures in several ways. Firstly, we observed the microphotographs of the cross section to characterize the nanostructure. Secondly, we examined the reaction products in a liquid phase using ICP-OES and TOC-TN methods, while vapor-phase products were examined by gas chromatography. Finally, according to the product data analysis, we demonstrate a mechanism for the electrochemical oxidation of n-GaN in oxalic acid, which involves 6 electrons.


Soluble inflammatory markers as predictors of liver histological changes in patients with chronic hepatitis C virus infection.
Host immune response seems to be mainly responsible for the progression of liver disease among patients with hepatitis C virus (HCV) infection. Immune activation involves the release of cytokines and their receptors that can be measured in plasma samples. The study aimed to evaluate the association between plasma levels of chemokines and soluble tumor necrosis factor receptors (sTNFR) and liver histological changes among patients with chronic HCV infection. Seventy-one treatment-naive patients were included. Plasma levels of CCL2, CCL3, CCL11, CCL24, CXCL9, CXCL10, sTNFR1, and sTNFR2 were measured and liver histological findings were reviewed. Plasma levels of CXCL9, sTNFR1, and sTNFR2 were significantly associated with liver fibrosis, with higher median levels found among patients with moderate/severe fibrosis (F >or= 2) if compared to those with no or mild fibrosis (p = 0.014; p = 0.012; p = 0.009, respectively). Plasma sTNFR2 levels were significantly associated with necroinflammatory activity, with higher median levels among patients with moderate/severe activity (A >or= 2) if compared to those with no or mild activity (2.34 ng/mL vs. 1.99 ng/mL; p = 0.019). In conclusion, plasma levels of CXCL9, sTNFR1, and sTNFR2 were independently associated with liver histological changes, suggesting a role of TNF activation and Th1-type cell-mediated immune response in the pathogenesis of HCV infection.


Self-supported heterogeneous titanium catalysts for enantioselective carbonyl-ene and sulfoxidation reactions.
A new strategy for the heterogenization of chiral titanium complexes was developed by the in situ assembly of bridged multitopic BINOL ligands with [Ti(OiPr)4] without using a support. The assembled heterogeneous catalysts (self-supported) showed excellent enantioselectivity in both the carbonyl-ene reaction of alpha-methylstyrene with ethyl glyoxylate (up to 98 % ee) and the oxidation of sulfides (up to >99 % ee). The catalytic performance of these heterogeneous catalytic systems was comparable or even superior to that attained with their homogeneous counterparts. The spacers between the two BINOL units of the ligands in the assembled catalysts had significant impact on the enantioselectivity of the carbonyl-ene reaction. This demonstrates the importance of the supramolecular structures of the assemblies on their catalytic behavior. In the catalysis of sulfoxidation, the self-supported heterogeneous titanium catalysts were highly stable and could be readily recycled and reused for over one month (at least eight cycles) without significant loss of activity and enantioselectivity (up to >99.9 % ee). The features of these self-supported catalysts, such as facile preparation, robust chiral structure of solid-state catalysts, high density of the catalytically active units in the solids, as well as easy recovery and simple recycling, are particularly important in developing methods for the synthesis of optically active compounds in industrial processes.


Psychopathy in a noninstitutional sample: differences in primary and secondary subtypes.
Early theoretical conceptualizations suggest psychopathy is a heterogeneous construct whereby psychopathic individuals are found in diverse populations. The current study examined male and female psychopathy subtypes in a large sample of undergraduate students (n = 1229). Model-based cluster analysis of the Psychopathic Personality Inventory-Short Form (PPI-SF) revealed two clusters in both male and female students. In males, the primary subtype evidenced greater psychopathic personality traits (i.e., Social Potency, Fearlessness, and Impulsive Nonconformity) and lower anxiety (i.e., higher Stress Immunity), whereas the secondary subtype displayed fewer psychopathic personality traits (i.e., Machiavellian Egocentricity and Blame Externalization) and higher anxiety (i.e., lower Stress Immunity). In females, the primary subtype exhibited higher scores across all PPI-SF subscales and lower anxiety whereas the secondary subtype reported lower PPI-SF subscale scores and higher anxiety. Across a diverse array of personality, affective, and behavioral external correlates, differences between the subtypes and with nonpsychopaths emerged. Implications for psychopathy in noninstitutional populations with respect to theory, research, and gender are discussed.


Graphene oxide decorated with gold enables efficient biophotovolatic cells incorporating photosystem I.
This paper describes the use of reduced graphene oxide decorated with gold nanoparticles as an efficient electron transfer layer for solid-state biophotovoltic cells containing photosystem I as the sole photo-active component. Together with polytyrosine-polyaniline as a hole transfer layer, this device architecture results in an open-circuit voltage of 0.3 V, a fill factor of 38% and a short-circuit current density of 5.6 mA cm-2 demonstrating good coupling between photosystem I and the electrodes. The best-performing device reached an external power conversion efficiency of 0.64%, the highest for any solid-state photosystem I-based photovoltaic device that has been reported to date. Our results demonstrate that the functionality of photosystem I in the non-natural environment of solid-state biophotovoltaic cells can be improved through the modification of electrodes with efficient charge-transfer layers. The combination of reduced graphene oxide with gold nanoparticles caused tailoring of the electronic structure and alignment of the energy levels while also increasing electrical conductivity. The decoration of graphene electrodes with gold nanoparticles is a generalizable approach for enhancing charge-transfer across interfaces, particularly when adjusting the levels of the active layer is not feasible, as is the case for photosystem I and other biological molecules.


Pulmonary complications in pediatric cardiac surgery at a university hospital.
To identify the prevalence of pulmonary complications in children undergone cardiac surgery, as well as demographic and clinical characteristics of this population. The sample comprised 37 children of both genders, underwent cardiac surgery at the Hospital Universitário Presidente Dutra, São Luis (MA) during the year of 2007. There were not included patients who had lung disease in pre-operative period, patients with neurological disorders, intra-operative death besides lack of data in medical records. The data were obtained from general medical and nursing staff of their medical records. The population of the study was predominantly composed by female children, from the countryside and at school age. Pathologies considered low risk were the majority, especially the patent ductus arteriosus, interventricular communication and interatrial communication. It was observed that the largest share of children made use of cardiopulmonary bypass for more than 30 minutes, with a median of 80 minutes, suffered a median sternotomy, using only the mediastinal drain and made use of mechanical ventilation after surgery, with the median about 6.6 hours. Only three (8.1%) patients developed pulmonary complications, and of these, two died. Most of the sample was female, school aged and from the countryside. The low time of cardiopulmonary bypass and mechanical ventilation, and congenital heart disease with low risk, may have been factors that contributed to the low rate of pulmonary complications postoperative.


Vasopressin mRNA regulation in individual hypothalamic nuclei: a northern and in situ hybridization analysis.
The present study examines the relative levels of vasopressin (AVP) mRNA within the paraventricular (PVN), supraoptic (SON), and suprachiasmatic (SCN) nuclei of the rat hypothalamus, and details the rates at which these levels change over the course of a 6 d salt-loading regimen. The quantitation of vasopressin mRNA was achieved by using three different procedures: (1) cell-free translation in rabbit reticulocyte lysate or (2) Northern analysis of poly(A)RNAs isolated from micro-punch dissected SON, PVN, and SCN, and (3) in situ hybridization histochemistry. The former involved the quantitative immunoprecipitation of the neurophysin precursors containing arginine8-vasopressin (AVP) or oxytocin, and the latter two techniques employed a radiolabeled synthetic oligodeoxynucleotide complementary to the 3' region of the AVP mRNA. Both the cell-free studies and the Northern gel analyses detected a sevenfold increase of AVP mRNA in the SON, a fivefold increase in the PVN, and no significant change in the SCN following 6 d of salt-loading. After the initiation of salt-drinking, these increases were shown to occur between 24 and 48 hr in the SON and between 48 and 72 hr in the PVN. The in situ hybridization studies revealed the anatomically correct hybridization of either 32P- or 3H-labeled AVP oligonucleotide to magnocellular perikarya within both the SON and PVN. Autoradiographic grains could be shown to be confined to the cytoplasm of these cells, and could be co-localized with immunoreactivity directed against the carboxy terminus of the AVP percursor. Comparison of x-ray level autoradiograms of control and 6 day salt-loaded SON revealed up to a sevenfold increase in specific signal in the salt-loaded sections. It is concluded that the response of AVP mRNA to osmotic stimuli in the three hypothalamic nuclei is heterogeneous, and that this heterogeneity can be explained by separating AVP neurons into two systems: one responsible for eliciting the antidiuretic actions of AVP via plasma AVP levels, and the other involved in CNS activities not directly involved with antidiuresis.


Biosynthesis of soluble carnitine acetyltransferases from the yeast Candida tropicalis.
Soluble carnitine acetyltransferase from Candida tropicalis is synthesized as a 76 kDa precursor, which is monomeric and possesses no or very little carnitine acetyltransferase activity. Maturation of the enzyme begins with proteolytic processing of the 76 kDa precursor to 64 and 57 kDa subunits. The processed subunits subsequently associate into two kinds of active oligomers; the 57 kDa subunits are assembled into a tetramer and the 64 kDa subunits into an octamer. Formation of these oligomers depends apparently on growth conditions, since both oligomers were present in cells grown in continuous culture, but cells grown batchwise contained only the tetrameric form of carnitine acetyltransferase.


In renal hypertension, Cirsium japonicum strengthens cardiac function via the intermedin/nitric oxide pathway.
Cirsium japonicum, a constituent of traditional Chinese medicine, has been shown to exert inflammatory effects as well as to improve the circulation and thus to counteract hematologic stasis. Studies have demonstrated that intermedin (IMD) has protective effects on hypertension in rats by regulating the Ang/NO metabolic pathway. In this study, we investigated whether by regulating the expression of IMD, Cirsium japonicum could improve cardiac function in rats with 2k1c-induced renal hypertension. Renal hypertension was induced in Sprague-Dawley rats by occluding the renal artery. The rats were maintained on a normal diet and randomly divided into four groups: sham, 2k1c, 2k1c with Cirsium japonicum (1.8 g/kg per day) and 2k1c with IMD (n = 10 in each group). Cardiac function, plasma angiotensin II (Ang II), IMD, serum nitric oxide (NO) and nitric oxide synthase (NOS), as well as the expression of IMD and adrenomedullin (ADM) in the aorta and left ventricle were analyzed. Administration of Cirsium japonicum or IMD significantly strengthened cardiac function in 2k1c-induced rats, increased serum NO and NOS levels, reduced plasma Ang II, and upregulated IMD expression in the aorta and left ventricle. These results demonstrate that Cirsium japonicum has cardioprotective effects on 2k1c-induced renal hypertension in rats via the IMD/NO pathway.


Overdose with modified-release paracetamol results in delayed and prolonged absorption of paracetamol.
A modified-release formulation of paracetamol is currently available in Australasia and marketed under a number of different trade names. These include: Panadol Osteo, Panadol Extend Tablets, and Duatrol SR. We report four cases of intentional overdose with this formulation resulting in delay to peak plasma paracetamol concentrations and prolonged paracetamol absorption. Physicians must be aware that a single plasma paracetamol estimation four or more hours post-ingestion may not be adequate in the risk assessment of patients requiring treatment with N-acetylcysteine (NAC). Current Australasian guidelines for the management of modified-release paracetamol overdose advise empiric commencement of NAC if the suspected ingested dose is greater than 10 grams or 200 mg/kg (whichever is the least), an initial plasma paracetamol concentration should be assayed four or more hours post-ingestion and a second assay should be estimated four hours after the first. Treatment with NAC should continue if either concentration falls above the paracetamol treatment nomogram line. With massive ingestions of this paracetamol formulation (>50 grams) plasma concentrations may be elevated for several days and prolonged treatment with NAC is recommended. When modified-release paracetamol overdose is suspected a clinical toxicologist or Poisons Information Centre should be consulted to help guide management decisions.


Food Proteins as Source of Opioid Peptides-A Review.
Traditional opioids, mainly alkaloids, have been used in the clinical management of pain for a number of years but are often associated with numerous side-effects including sedation, dizziness, physical dependence, tolerance, addiction, nausea, vomiting, constipation and respiratory depression which prevent their effective use. Opioid peptides derived from food provide significant advantages as safe and natural alternative due to the possibility of their production using animal and plant proteins as well as comparatively less side-effects. This review aims to discuss the current literature on food-derived opioid peptides focusing on their production, methods of detection, isolation and purification. The need for screening more dietary proteins as a source of novel opioid peptides is emphasized in order to fully understand their potential in pain management either as a drug or as part of diet complementing therapeutic prescription.


Pseudomonas aeruginosa cytochrome P450 CYP168A1 is a fatty acid hydroxylase that metabolizes arachidonic acid to the vasodilator 19-HETE.
Pseudomonas aeruginosa is a Gram-negative opportunistic human pathogen that is highly prevalent in individuals with cystic fibrosis (CF). A major problem in treating CF patients infected with P. aeruginosa is the development of antibiotic resistance. Therefore, the identification of novel P. aeruginosa antibiotic drug targets is of the utmost urgency. The genome of P. aeruginosa contains four putative cytochrome P450 enzymes (CYPs) of unknown function that have never before been characterized. Analogous to some of the CYPs from Mycobacterium tuberculosis, these P. aeruginosa CYPs may be important for growth and colonization of CF patients' lungs. In this study, we cloned, expressed, and characterized CYP168A1 from P. aeruginosa and identified it as a subterminal fatty acid hydroxylase. Spectral binding data and computational modeling of substrates and inhibitors suggest that CYP168A1 has a large, expansive active site and preferentially binds long chain fatty acids and large hydrophobic inhibitors. Furthermore, metabolic experiments confirm that the enzyme is capable of hydroxylating arachidonic acid, an important inflammatory signaling molecule present in abundance in the CF lung, to 19-hydroxyeicosatetraenoic acid (19-HETE; Km = 41 μM, Vmax = 220 pmol/min/nmol P450), a potent vasodilator, which may play a role in the pathogen's ability to colonize the lung. Additionally, we found that the in vitro metabolism of arachidonic acid is subject to substrate inhibition and is also inhibited by the presence of the antifungal agent ketoconazole. This study identifies a new metabolic pathway in this important human pathogen that may be of utility in treating P. aeruginosa infections.


Validation of Doloplus-2 among nonverbal nursing home patients--an evaluation of Doloplus-2 in a clinical setting.
Pain measurement in nonverbal older adults is best based on behavioural observation, e.g. using an observational measurement tool such as Doloplus-2. The purposes of this study were to examine the use of Doloplus-2 in a nonverbal nursing home population, and to evaluate its reliability and validity by comparing registered nurses' estimation of pain with Doloplus-2 scores. In this cross-sectional study, Doloplus-2 was used to observe the pain behaviour of patients aged above 65 years who were unable to self-report their pain. Nurses also recorded their perceptions of patient pain (yes, no, don't know) before they used Doloplus-2. Data on demographics, medical diagnoses, and prescribed pain treatment were collected from patient records. Daily life functioning was measured and participants were screened using the Mini Mental State Examination. In total, 77 nursing home patients were included, 75% were women and the mean age was 86 years (SD 6.6, range 68-100). Over 50% were dependent on nursing care to a high or a medium degree, and all were severely cognitively impaired. The percentage of zero scores on Doloplus-2 ranged from 17% (somatic reactions) to 40% (psychosocial reactions). Cronbach's alpha was 0.71 for the total scale. In total, 52% of the patients were judged by nurses to be experiencing pain, compared with 68% when using Doloplus-2 (p = 0.01). For 29% of the sample, nurses were unable to report if the patients were in pain. In the present study, more patients were categorized as having pain while using Doloplus-2 compared with nurses' estimation of pain without using any tools. The fact that nurses could not report if the patients were in pain in one third of the patients supports the claim that Doloplus-2 is a useful supplement for estimating pain in this population. However, nurses must use their clinical experience in addition to the use of Doloplus-2, as behaviour can have different meaning for different patients. Further research is still needed about the use of Doloplus-2 in patients not able to self-report their pain.


Schistosoma mansoni-Induced Oxidative Stress Triggers Hepatocellular Proliferation.
Schistosomiasis is one of the most prominent parasite-induced infectious diseases, affecting more than 250 million people. Schistosoma mansoni causes metabolic exhaustion and a strong redox imbalance in the liver, causing parenchymal damage, and may predispose for cancer. We investigated whether oxidative stress provokes hepatocellular proliferation upon S. mansoni infection. The cell cycle, replication stress response, and proliferation were analyzed on transcriptional and protein levels in the livers of S. mansoni-infected hamsters and by mechanistic gain- and loss-of-function experiments in human hepatoma cells. Major results were validated in human biopsy specimens of S. mansoni-infected patients. S. mansoni infection induced licensing factors of DNA replication and cell-cycle checkpoint cyclins in parallel with a DNA damage response in hamster hepatocytes. Moreover, even unisexual infection without egg effects, as a reflection of a chronic inflammatory process, resulted in a moderate activation of several cell-cycle markers. S. mansoni soluble egg antigens induced proliferation of human hepatoma cells that could be abolished by reduced glutathione. Our data suggest that hepatocellular proliferation is triggered by S. mansoni egg-induced oxidative stress.


MAP kinase-mediated phosphorylation of distinct pools of histone H3 at S10 or S28 via mitogen- and stress-activated kinase 1/2.
ERK and p38 MAP kinases, acting through the downstream mitogen- and stress-activated kinase 1/2 (MSK1/2), elicit histone H3 phosphorylation on a subfraction of nucleosomes--including those at Fos and Jun--concomitant with gene induction. S10 and S28 on the H3 tail have both been shown to be phospho-acceptors in vivo. Both phospho-epitopes appear with similar time-courses and both occur on H3 tails that are highly sensitive to TSA-induced hyperacetylation, similarities which might suggest that MSK1/2 phosphorylates both sites on the same H3 tails. Indeed, on recombinant histone octamers in vitro, MSK1 efficiently phosphorylates both sites on the same H3 tail. However, sequential immunoprecipitation studies show that antibodies against phosphorylated S10-H3 recover virtually all this epitope without depletion of phosphorylated S28-H3, and vice versa, indicating that the two phospho-epitopes are not located on the same H3 tail in vivo. Confocal immunocytochemistry confirms the clear physical separation of the two phospho-epitopes in the intact mouse nucleus. Finally, we used transfection-based experiments to test models that might explain such differential targeting. Overexpression and delocalisation of MSK1 does not result in the breakdown of targeting in vivo despite the fact that the ectopic kinase is fully activated by external stimuli. These studies reveal a remarkable level of targeting of S10 and S28 phosphorylation to distinct H3 tails within chromatin in the interphase mouse nucleus. Possible models for such exquisite targeting are discussed.


Immunoglobulin M antibodies to aquaporin-4 in neuromyelitis optica and related disorders.
Neuromyelitis optica (NMO, Devic syndrome) is an inflammatory disorder of the central nervous system of putative autoimmune etiology that primarily affects the optic nerves and spinal cord. NMO is frequently associated with immunoglobulin G (IgG) antibodies to aquaporin-4 (AQP4-IgG), which are thought to be involved in the patho-genesis of the disease. The frequency and diagnostic relevance of immunoglobulin M (IgM) antibodies to aquaporin-4 (AQP4-IgM) in patients with NMO is essentially not known. Testing for AQP4-IgM may be of importance since 20%-30% of patients with NMO are negative for AQP4-IgG. Moreover, IgM antibodies are more potent activators of complement compared with IgG, and are detectable at NMO lesional sites. Serum samples from 42 patients with NMO spectrum disorders (NMOSD) and from 66 controls were tested for IgM AQP4-Ab using a cell-based assay employing HEK293 cells transfected with human full length AQP4. To control for possible interactions between IgG and IgM, serum was depleted of IgG prior to testing by indirect immunofluorescence. IgM AQP4-Ab were detectable in 4/42 samples from patients with NMOSD, but in none of the 66 control samples. In three patients, titers were higher following depletion of total IgG from the samples. One sample was positive only after precipitation of total IgG. AQP4 antibodies of the IgM class exist in almost 10% of patients with NMO and might contribute to lesion pathology. Routine testing for AQP4-IgM appears to not be justified, as all AQP4-IgM positive patients were also positive for AQP4-IgG, and none of the AQP4-IgG negative samples were positive for AQP4-IgM.


Sex differences in elbow and wrist joint loading during the cartwheel and round off with different hand positions performed by young gymnasts.
The aim of the study was to determine if sex differences exist in the key elbow and wrist joint injury risk factors during different cartwheel (CW) and round-off (RO) techniques performed by young male and female artistic gymnasts. Sixteen active young gymnasts (8 males and 8 females) performed 30 successful trials of CW and RO with three different hand positions (parallel (10), T-shape (10) and reverse (10)). Synchronised kinematic and kinetic data were collected for each trial. Two-way repeated measures ANOVA (3 × 2, technique × sex) and effect-sizes (ES) were used for statistical analysis. In conclusion, female gymnasts exhibited greater normalised peak vertical ground reaction forces (VGRF), elbow and wrist compression forces and elbow internal adduction moments during CW and RO skills compared with male gymnasts. In both sexes, the parallel and reverse techniques increased peak VGRF, elbow and wrist compression forces and the elbow internal adduction moment. Increased elbow flexion resulted in decreased peak VGRF, elbow compression forces and elbow internal adduction moment. Injury risk factors including elbow extension and internal adduction moment with axial compression force suggest that a CW and RO in reverse and parallel techniques can be hazardous especially for young female gymnasts.


Metastatin: a hyaluronan-binding complex from cartilage that inhibits tumor growth.
In this study, a hyaluronan-binding complex, which we termed Metastatin, was isolated from bovine cartilage by affinity chromatography and found to have both antitumorigenic and antiangiogenic properties. Metastatin was able to block the formation of tumor nodules in the lungs of mice inoculated with B16BL6 melanoma or Lewis lung carcinoma cells. Single i.v. administration of Metastatin into chicken embryos inhibited the growth of both B16BL6 mouse melanoma and TSU human prostate cancer cells growing on the chorioallantoic membrane. The in vivo biological effect may be attributed to the antiangiogenic activity because Metastatin is able to inhibit the migration and proliferation of cultured endothelial cells as well as vascular endothelial growth factor-induced angiogenesis on the chorioallantoic membrane. In each case, the effect could be blocked by either heat denaturing the Metastatin or premixing it with hyaluronan, suggesting that its activity critically depends on its ability to bind hyaluronan on the target cells. Collectively, these results suggest that Metastatin is an effective antitumor agent that exhibits antiangiogenic activity.


Recombination of pseudorabies virus vaccine strains in swine.
We report here genetic recombination between 2 USDA-licensed vaccine strains of pseudorabies virus co-inoculated into swine. The vaccine strains, one of which was a conventionally attenuated strain and the other, a genetically engineered deleted strain containing a negative immunologic marker, had complementary genomes. Co-inoculation resulted in the creation of novel strains of pseudorabies virus containing negative immunologic markers with restored virulence genes. Plaque-purified recombinant progeny viruses were found in 2 litters of pigs in which both strains were co-inoculated IM, a litter in which both strains were co-inoculated oronasally, and a litter in which the conventionally attenuated strain was inoculated oronasally and the genetically engineered strain was inoculated IM. Recombinant phenotypes and recombinant restriction fragment patterns were observed. The creation, spread, and potential misdiagnosis of these types of recombinant strains could disrupt control and eradication programs that are based on the serologic identification of swine infected with potentially virulent strains of pseudorabies virus.


Genome engineering with TAL-effector nucleases and alternative modular nuclease technologies.
Over three years following the discovery of the TAL code, artificial TAL effector DNA binding domains have emerged as the premier platform for building site-specific DNA binding polypeptides for use in biological research. Here, we provide an overview of TAL effector and alternative modular DNA binding domain (mDBD) technologies, focusing on their use in established and emerging architectures for building site-specific endonucleases for genome engineering applications. We also discuss considerations for choosing TAL effector/mDBD or alternative nuclease technologies for genome engineering projects ranging from basic laboratory gene editing of cultured cell lines to therapeutics. Finally, we highlight how the rapid pace of development of mDBD-based, such as monomeric TALENs (I-TevI-TAL), and more recently RNA-guided nucleases (CRISPR-Cas9) has led to a transition in the field of genome engineering towards development of the next generation of technologies aimed at controlling events that occur after targeted DNA breaks are made.


ICBT program for improving informal caregiver well-being: A qualitative study.
Informal caregivers represent an important and vulnerable part of the society. They can experience negative psychological symptoms, such as depression and anxiety. Internet-based cognitive behavioural therapy (ICBT) is a promising psychological support option that could be effective in reducing informal caregiver burden as well as improving psychological well-being. Qualitative studies are valuable for gaining insights into participant experiences of using ICBT. The main aim of this study was to explore informal caregiver experiences and hence acceptability and feasibility of the transdiagnostic ICBT program aimed at reducing informal caregiver burden and increasing the quality of life. Following a strategic sampling procedure, 23 informal caregivers were recruited from previous randomized controlled trial for informal caregivers in Lithuania. Participants were interviewed over phone, using semi-structured interview questions. Interviews were analysed using a thematic analysis approach. Four themes and a total of 10 sub-themes were generated: I A program as a means of change (Convenience and applicability of the format and materials; Ability to focus on own needs; Opportunity for communication), II Suggestions for the program (Including live support; Tailoring materials and format; Providing with more time and resources), III Driving personal and situational forces (Developing acceptance and adjustment over time; Being proactive and/or receiving support) and Hindering situational factors (Deterioration and unpredictability of the care-receivers health; Lacking external support and opportunities for respite). Most of the informal caregivers were found to be satisfied with the program's format and materials as well as the communication with the therapist via a message function in the program. Some suggestions were made regarding implementation of the live support option. Also, suggestions regarding possibility for tailoring the program's content. Lastly, several personal and situational factors were identified as important in affecting informal caregiver well-being. We conclude that ICBT has potential in reducing informal caregiver burden and improving psychological health. Further research trials are warranted for evaluating both, the effectiveness and the feasibility of the program.


Environmental risk assessment for ancillary substances in biotechnological production of pharmaceuticals.
An increasing number of pharmaceutical active substances are produced through biotechnological processes. For sustained and safe growth of the host organisms as well as optimal expression, purification, and formulation of the product, biotechnological manufacturing processes need optimal and robust environmental conditions, which are attained through the use of buffers, chelators, and antibiotics, beside nutrients. These ancillary substances are drained with the wastewater to a wastewater treatment plant (WWTP) and are released after treatment with the effluent to receiving waters. The potential risks of such substances to WWTPs and surface waters were investigated. Three common buffers (morpholinoethane sulfonic acid [MES], morpholinopropanesulfonic acid [MOPS], 1,4-piperazine (diethanesulfonic acid) [PIPES]), one chelator (ethylenediaminetetraacetic acid [EDTA]), and one antibiotic (gentamycin) were searched in the literature for environmental data or tested for biodegradability and inhibition of activated sludge as well as acute toxicity to algae, daphnids, and fish. Amounts of the ancillary substances used in the European biotechnological production plants of F. Hoffmann-La Roche Ltd in Basle (Switzerland) and Penzberg (Germany), and actual wastewater fluxes through the respective WWTP, as well as realistic dilution factors for the local receiving water, were documented. Based on this information, site-specific predicted environmental concentrations (PECs) for the WWTPs and surface waters in Basle and Penzberg were extrapolated. These PECs were compared with predicted no effect concentrations (PNECs) for the WWTP and surface waters, derived from sludge inhibition and ecotoxicity results, respectively. For all five ancillary substances investigated, all PEC/PNEC risk characterization ratios are <1, indicating no significant risk to the WWTPs or the receiving waters at both sites.


Antagonism of synaptic transmission in vivo: contributions of microiontophoresis.
Fundamental to the problems in the study of synaptic transmission in the mammalian central nervous system is the identification of the nature and role of transmitter substances at prescribed loci. Although recently many advances have been made in determining which compounds are endogenous to the central nervous system and which of these and other molecules alter neuronal activity, there can be little doubt that at the present level of our understanding of such brain constituent we are encountering merely a few of the basic materials requisite for the normal functioning of central neurones. Some 60 or 70 substances have now been postulated as candidates for involvement in synaptic processes. Many of the receptors for these and closely related compounds have been characterized pharmacologically through the examination of the effects of agonists and antagonists. One technique which has played a principal role in these neuropharmacological investigations is the method of local drug administration termed microiontophoresis. Although this method is invaluable for helping to answer questions at the molecular and membrane level, under circumscribed conditions its advantages may be exploited for use in experiments oriented toward a more physiological level of investigation. This short account is an attempt to provide a description of some of these conditions and further will describe some of the creative ways in which microiontophoresis has been utilized in vivo to reveal mechanisms by which the central nervous system processes information at the synaptic level.