Improving patient care by virtual case discussion between plastic surgeons and residents of Uganda and the Netherlands.
Traditional on-site missions of plastic surgeons from "high-income countries" in "low- and middle-income countries" are often limited in time and lack proper follow-up. Regular digital collaboration could lead to a more impactful and durable exchange of knowledge for plastic surgeons and residents in both settings. The aim of this study was to evaluate the satisfaction of the first twelve months of weekly digital meetings, explore advantages/disadvantages, and to provide tools for similar initiatives. Weekly meetings started from August 2021. An encrypted digital connection allowed residents and plastic surgeons from Uganda and the Netherlands to discuss cases for educational purposes, where treatment options were considered. After twelve months, a survey was sent to participants from both countries to indicate the meetings' strengths, weaknesses, and possible improvements. A total of 18 participants responded to the questionnaire (ten plastic surgeons, six residents, and two researchers). The strengths of the meetings were the accessibility of the meetings, knowledge exchange and practice for residents' final exams. Possible improvements included having a clear format for patient discussion, a session moderator and better internet connectivity. Moreover, a database to assess the impact of the given intervention on the patient cases by evaluating postoperatively (e.g. three months), could further improve clinical care. Virtual patient discussions subjectively contributed to medical education at both locations. Improved digital infrastructure and a collaborative database could further maximize learning capacity. Furthermore, digital proctoring is a promising way to establish sustainable collaborations between high- and low-resource countries.


Spatial Regulation of Valve Interstitial Cell Phenotypes within Three-Dimensional Micropatterned Hydrogels.
Calcific aortic valve disease (CAVD) is the third leading cause of cardiovascular disease. CAVD exhibits progressive disruption of the normally highly organized and aligned extracellular matrix (ECM) structure within the valve leaflets simultaneously with myofibroblastic and/or osteogenic differentiation of indigenous endogenous valve interstitial cells (VIC). It is unclear how the alignment of VIC within their 3D microenvironment drives VIC phenotype or how alignment affects cellular responses to biochemical cues in physiological or pathological conditions. In this study, we implement a photolithographic technique to control the alignment and elongation of both normal and diseased human aortic VIC (HAVIC) within microengineered 3D hydrogels consisting of methacrylated hyaluronic acid and methacrylated gelatin. Stripe micropatterning created distinct alignment of HAVIC within a 3D culture system, which promoted spreading and enhanced their activation and osteogenic differentiation in pro-osteogenic conditions. HAVIC from a patient with CAVD exhibited greater susceptibility to myofibroblastic and osteogenic differentiation in culture. The roles of conjugated basic fibroblastic growth factor (bFGF) and RhoA/ROCK pathway in regulating HAVIC phenotypes were also investigated in the presence of aligned microtopography. The addition of bFGF was preventative to osteogenic differentiation for healthy HAVIC; however, it promoted osteogenic differentiation in diseased HAVIC. Inhibition of the ROCK pathway only decreased osteogenic differentiation for diseased HAVIC in the aligned formation. Collectively, these results improve our knowledge of the effects that VIC alignment has on VIC phenotypes and valve disease progression. The cell culture platform also enables a better understanding of the interplay between topography, biochemical cues, and VIC differentiation and provides information useful for directing differentiation as well as valve tissue regeneration.


Density Functional Theory and Raman Spectroscopy Studies of Adsorption Sites of Au Nanoparticles with Alectinib.
Alectinib is an ALK tyrosine kinase inhibitor, which is mainly used in patients with crizotinib-resistant nonsmall cell lung cancer. Alectinib has attracted much clinical attention for its longest progression-free survival time and the best therapeutic effect. The chemical adsorption of Au nanoclusters (AuNPs) with alectinib molecules is studied by density functional theory (DFT) and surface-enhanced Raman scattering spectroscopy (SERS) experiments. DFT/B3LYP-D3/6-311G** was used for optimization and vibration analysis of alectinib-Au6 complexes, as well as molecular electrostatic potential, frontier molecular orbital, and electro-optic-based charge transfer descriptors. Comparing the results of the DFT theory and SERS experiment, alectinib and AuNPs can form Au-N6 bonds primarily through chemical adsorption of N6 atoms, and the experimental results showed that the enhancement factor (EFCHEM) could reach 4.27. The results provide a theoretical basis for exploring the mechanism of chemical enhancement between AuNPs and alectinib.


Endovascular therapy yields significantly superior outcomes for large vessel occlusions compared with intravenous thrombolysis: is it time to randomize?
We compared outcomes between endovascular (EV) therapy and intravenous (IV) thrombolysis in large vessel strokes. 223 patients who had received either IV (n=100) or EV (n=123) therapy were analyzed. Only patients with strokes involving the internal carotid artery terminus (ICA-T, n=45), the middle cerebral artery (M1, n=107) or the bifurcation branches (M2, n=71) were included. The primary endpoint was 3 month outcome based on the modified Rankin Scale (mRS) score, good-outcome defined as mRS ≤2. The good outcome was 44.7% in the EV group and 26% in the IV group (p=0.003, OR 2.3, 95% CI 1.3 to 4.1). There was no difference in mortality or hemorrhage. For ICA-T occlusions, the good outcome was 27.6% in the EV and 0% in the IV group (p=0.004); for M1 occlusions, 40.6% in the EV versus 10.5% in the IV group (p=0.0006, OR 5.8, 95% CI 1.9 to 18.2); and for M2 occlusions, 76% in the EV versus 47.8% in the IV group (p=0.01, OR 3.5, 95% CI 1.2 to 10.2). For M1 occlusions, the death rate was 27.5% for the EV compared with 57.9% for the IV group (p=0.002, OR 3.6, 95% CI 1.6 to 8.3) with no difference observed in mortality for ICA-T or M2 occlusions. In the univariate analysis, age, National Institutes of Health Stroke Scale score and occlusion site were significant predictors of outcome and mortality (p<0.0001 for all). In the multivariable analysis, EV therapy (p=0.0004, OR 3.9, 95% CI 1.8 to 9) and younger age (p<0.0001, OR 0.96, 95% CI 0.9 to 0.98) were significant independent predictors of good outcome. There are significantly higher odds of a favorable outcome with EV compared with IV therapy for large vessel strokes. The data support the rationale of a randomized trial for large vessel occlusions.


Evaluation of herpes simplex detection in corneal scrapings by three molecular methods.
Herpes simplex virus (HSV)-1 keratitis (HSK) is a sight-threatening ocular infection with worldwide occurrence. A prompt laboratory diagnosis is often very useful. The purpose of this study was to evaluate molecular methods as rapid diagnostic tools compared with cell culture of HSK. Corneal scrapings from patients with clinically suspected HSK were tested by direct immunofluorescence assay (IFA) for HSV-1 antigen and by polymerase chain reaction (PCR) for HSV-1 DNA, and an attempt for viral isolation was performed on Vero cell line culture. Positive samples by cell culture were 20.8%, whereas PCR was positive in 29.2%, and IFA was positive in 33.3%. IFA had better sensitivity (80%) and negative predictive value (81.8%) than PCR (70% and 76.9%, respectively); however, PCR had better specificity (71.4%) and positive predictive value (63.6%). This indicates that a combination of cell culture, IFA and PCR constitutes the best set of tools for diagnosis of clinically suspected cases of HSK. Documented infection can be further assessed by cell-culture technique or PCR depending laboratory availability.


Differences in wedge factor determination in air using a PMMA mini-phantom or a brass build-up cap.
The head scatter dose contribution to the output of a treatment machine has been determined for an open and wedged 60Co gamma-ray beam and for open and wedged x-ray beams of 4, 8, and 16 MV. From those data wedge factor values "in air" have been deduced, expressed as the ratio of the dose to water, measured in air, for the situation with and without wedge, for the same number of monitor units (or treatment time for 60Co). The measurements have been performed using a polymethyl-metacrylate (PMMA) and a graphite-walled ionization chamber inserted in a brass build-up cap and in a PMMA mini-phantom, respectively. Absolute wedge factor values deduced with both detector systems and based on the ratio of ionization chamber readings, differ for the investigated photon beams, up to 3.5% for the 4 MV x-ray beam. The deviations results from the difference in composition between the detector materials and water and can be taken into account by conversion of the ionization chamber readings for both the open and wedged photon beams to the absorbed dose to water. For the brass build-up cap detector system the ratio of the conversion factors for the wedged and open beam changes the ratio of the ionization chamber readings up to about 3.6% for the 4 MV x-ray beam. For the mini-phantom the conversion factors for the wedged and open beam are almost equal for all photon beams. Consequently, for that system wedge factors based on ionization chamber readings or dose values are the same. With respect to the wedge factor variation with field size a somewhat larger increase has been determined for the 60Co and 4 MV photon beam using the brass build-up cap: about 1% for field sizes varying between 5 cm x 5 cm and 15 cm x 15 cm. This effect has to be related to an apparent more pronounced variation of the head scatter dose contribution with field size for the wedged photon beams if the brass build-up cap detection system is used. It can be concluded that determination of wedge factors "in air" under reference irradiation conditions, performed with both the mini-phantom and brass build-up cap yields within 0.5% the same result if the wedge factors are based on a dose to water ratio. However, by using high-Z build-up materials the determination is more complicated because appropriate conversion factors are then required, while similar conversion factors can be ignored if more water equivalent build-up materials such as PMMA are applied.


Early infant diagnosis of HIV infection in southeastern Nigeria: prevalence of HIV infection among HIV-exposed babies.
Vertical transmission of HIV-1 is responsible for a high level of infant mortality necessitating early infant diagnosis. Serologic tests are not useful because of persistence of maternal antibodies in infants. Amplification of the integrated viral genome by PCR is the preferred method of diagnosis of HIV infection in these children. To determine the prevalence of HIV among a cohort of HIV-exposed babies. HIV-exposed infants were recruited for DNA PCR (early infant diagnosis). Babies were enrolled from six weeks of age. Relevant data were collected with the aid of a proforma. Mothers were given pre-test counselling. Heel or finger prick samples of blood on Whatman filter paper were used for DNA PCR testing. Data on the initial 304 babies enrolled for DNA PCR were analyzed. Seven (3.6%) of 192 mother-baby pairs who had received requisite prophylactic anti-retrovirals (PARV) were PCR-positive. In 23 (8.7%) PCR positive babies, their mothers received PARV but the babies had no post exposure prophylaxis (PEP), while two (12.5%) of 16 babies who had received PARV without their mothers turned out PCR-positive. Thirty nine (53.4%) of 73 mother-baby pairs who had no PARV were infected. Exclusive breastfeeding (EBF) rate was 35.5%. In these babies five (18.5%) were infected, while 288 (75%) of babies were exclusive formula fed (EFF), out of which 11 (4.8%) were infected. Forty-seven (15.5%) of the babies were mixed-fed, and 32 (68.0%) of them were infected. Prophylactic ARV in mothers and babies gave a marked reduction in Mother-to-Child-Transmission (MTCT) rate. Feeding BMS conferred a superior protection against (MTCT) than EBF.


Relation between severe illness and non-completion of quality-of-life questionnaires by patients with rectal cancer.
Quality of life (QoL) is an important outcome measure in clinical studies, but interpretation is hindered by incompleteness of data. We addressed this issue in a population-based cohort study of 146 patients with newly diagnosed rectal cancer. QoL was assessed by means of European Organization for the Research and Treatment of Cancer questionnaires at discharge from hospital after primary treatment and then every 3 months for 2 years. In parallel, objective clinical data were documented. Analyses were conducted in three steps: participants versus non-participants with QoL-assessment; poor compliers who filled in only one or two questionnaires (n=20) versus good compliers who filled in all or nearly all questionnaires (n=18); and the proportion of missing forms and critical (very poor) QoL scores in risk patients versus non-risk patients over the course of 2 years. Non-participants and poor compliers were older, were more likely to receive palliative (rather than curative) treatment, and had worse scores for physical status. Tumour progression and therapeutic interventions were more frequent in poor compliers than in good-compliers. Patients with risk factors (age 475 years, poor physical status, palliative treatment) were more likely to have missing questionnaires and critical QoL scores in respect of physical functioning and global quality of life over the course of 2 years. Missing values for QoL have clinical as well as methodological implications, because QoL scores can enhance a clinician's insight. Unwillingness to fill in a questionnaire is an indicator of serious illness. Studies that report sample statistics without specifying compliance rates and the characteristics of non-compliers will give a misleadingly positive picture.


Vitamin D supplementation and inflammatory and metabolic biomarkers in patients with knee osteoarthritis: post hoc analysis of a randomised controlled trial.
The aim of this study was to determine whether vitamin D supplementation and maintaining vitamin D sufficiency are associated with changes in inflammatory and metabolic biomarkers in patients with knee osteoarthritis (OA) and vitamin D deficiency. A total of 413 participants with symptomatic knee OA and vitamin D deficiency were enrolled in a randomised, placebo-controlled trial and received 1·25 mg vitamin D3 or placebo monthly for 24 months across two sites. In this post hoc analysis, 200 participants from one site (ninety-four from the placebo group and 106 from the vitamin D group; mean age 63·1 (sd 7·3) years, 53·3 % women) were randomly selected for measurement of serum levels of inflammatory and metabolic biomarkers at baseline and 24 months using immunoassays. In addition, participants were classified into two groups according to serum 25-hydroxyvitamin D (25(OH)D) levels at months 3 and 24: (1) not consistently sufficient (25(OH)D≤50 nmol/l at either month 3 or 24, n 61), and (2) consistently sufficient (25(OH)D>50 nmol/l at both months 3 and 24, n 139). Compared with placebo, vitamin D supplementation had no significant effect on change in serum high-sensitive C-reactive protein, IL-6, IL-8, IL-10, leptin, adiponectin, resistin, adipsin and apelin. Being consistently vitamin D sufficient over 2 years was also not associated with changes in these biomarkers compared with not being consistently sufficient. Vitamin D supplementation and maintaining vitamin D sufficiency did not alter serum levels of inflammatory and metabolic biomarkers over 2 years in knee OA patients who were vitamin D insufficient, suggesting that they may not affect systemic inflammation in knee OA patients.


Analysis and critical comparison of food allergen recalls from the European Union, USA, Canada, Hong Kong, Australia and New Zealand.
As part of a European Union-funded project (FP7) developing 'Integrated approaches to food allergen and allergy management', a database was constructed based on publicly available information on food allergen recalls in Europe, North America, Hong Kong, Australia and New Zealand. Over 2000 entries were made into the database. The database covers a 4-year period from 2011 to 2014 and each entry is categorised into food type (two different classifications), identified allergen and cause where indicated by the authorities. Across different authorities, by far the biggest incidence of undeclared allergens occurred in the food categories of prepared dishes and snacks (range = 12-53%), and cereals and bakery products (range = 14-25% of all recalls and/or alerts). The biggest incidence of undeclared allergens, according to the information from most authorities, occurred for milk and milk products (16-31% of all products with recall or alert), followed by cereals containing gluten (9-19%), soy (5-45%), and egg and egg products (5-17%). Although 42-90% of the products with recalls/alerts were explained as being 'Not indicated on the label', this is a generic explanation of cause and does not provide much insight into the causes of the recall/alerts. However, 0-17% of products with recalls/alerts could be coded as caused by the unintended presence of an allergen as the probable result of cross-contact in production. Construction of the database of allergen recalls has provided some important lessons and recommendations to the authorities are made in this paper in terms of the harmonisation of the reporting of allergen recalls into a more standardised format.


Nuclear and chloroplast DNA phylogeography reveals high genetic diversity and postglacial range expansion in Quercus mexicana.
Phylogeographical studies are fundamental for understanding factors that influence the spatial distribution of genetic lineages within species. Population expansions and contractions, distribution shifts, and climate changes are among the most important factors shaping the genetic compositions of populations. We investigated the phylogeography of an endemic oak, Quercus mexicana (Fagaceae), which has a restricted distribution in northeastern Mexico along the Sierra Madre Oriental and adjacent areas. Nuclear and chloroplast DNA microsatellite markers were used to describe the genetic diversity and structure of 39 populations of Q. mexicana along its entire distribution area. We tested whether population expansion or contraction events influenced the genetic diversity and structure of the species. We also modeled the historical distributional range of Q. mexicana (for the Mid Holocene, the Last Glacial Maximum, and the Last Interglacial) to estimate the extent to which climate fluctuations have impacted the distribution of this oak species. Our results revealed high genetic diversity and low genetic structure in Q. mexicana populations. Ecological niche models suggested historical fluctuations in the distributional range of Q. mexicana. Historical range changes, gene flow, and physical barriers seem to have played an important role in shaping the phylogeographic structure of Q. mexicana. Our study indicates that the genetic structure of Q. mexicana may have been the result of responses of oak trees not only to heterogeneous environments present in the Sierra Madre Oriental and adjacent areas, but also to elevational and latitudinal shifts in response to climate changes in the past.


Recent issues on dosimetry and radiobiology for peptide receptor radionuclide therapy.
Peptide receptor radionuclide therapy (PRRT) has been constantly evolving over the last decade, providing successful results in the treatment of tumors expressing somatostatin receptors, especially with 90Y -- and 177Lu -- radiolabelled peptides. Recent and/or ongoing studies assure new perspectives to come. Dosimetry represents a precious guide for the selection of radionuclides and peptides, for protocol settings, for toxicity prevention and therapy optimization. Thus, reliable and personalized dosimetry is more and more requested. This paper reviews the important advances recently obtained in the dosimetric methods that have been applied to this therapy. Special emphasis has been given to the impact derived (or derivable in the next future) from more refined dose evaluations focused on the kidneys and the red marrow. The possibility of improving the accuracy of dosimetry represents a further challenge for this therapy. Following the preliminary correlation observed between the biological effective dose and the probability of renal injury, more reliable dose estimates could definitively enhance the predicitivity of the radiobiological effects, for toxicity prevention as well as for tumor control.


The detection of any coronary calcium outperforms Framingham risk score as a first step in screening for coronary atherosclerosis.
The Framingham risk score is often recommended as the starting point for coronary disease screening. We compared the sensitivity of the Framingham risk score for moderate or greater degrees of atherosclerosis to the sensitivity achieved by simple observation of whether any coronary calcium is present. The reference standard was plaque burden as determined by coronary CT angiography. Of 1,416 men (mean age, 51.4 +/- 9.9 [SD] years) and 707 women (56.9 +/- 10.6 years), most were asymptomatic. Plaque burden (segment plaque score) and stenoses burden (Duke prognostic score) were estimated. A segment plaque score >or= 4 or a Duke prognostic score >or= 3 indicated moderate or greater disease burden. For a segment plaque score >or= 4, the presence of any calcium was 98% sensitive in men and 97% sensitive in women, whereas a Framingham risk score >or= 10% was 74% sensitive in men and 36% sensitive in women. The negative likelihood ratio for the presence of calcium was 0.04 in subjects of either sex, whereas, for a Framingham risk score <or= 10%, the negative likelihood ratio was 0.38 in men and 0.71 in women. For a Duke prognostic score >or= 3, calcium was 97% sensitive in men and 92% sensitive in women, whereas a Framingham risk score >or= 10% was 88% sensitive in men and 35% sensitive in women. The negative likelihood ratio of calcium presence was 0.05 in men and 0.13 in women, whereas the negative likelihood ratio for a Framingham risk score <or= 10% was 0.20 in men and 0.75 in women. If subjects are excluded from further screening because they are in the Framingham low-risk category, almost two thirds of women and a quarter of men with substantial atherosclerosis will be missed. In contrast, the simple observation of any coronary calcium is highly sensitive and moderately specific.


The role of bone modifying agents for secondary osteoporosis prevention and pain control in post-menopausal osteopenic breast cancer patients undergoing adjuvant aromatase inhibitors.
Hormonal therapy (HT) blocks the hormone-mediated growth signal dramatically reducing estrogenic levels with aromatase inhibitors (AIs) becoming a crucial component of the treatment mainstay in patients with early breast cancer (BC). Postmenopausal BC patients receiving HT present with a significant risk of secondary osteoporosis with AIs further reducing estrogen levels and ultimately leading to an accelerated rate of bone resorption and thus decreased bone mineral density (BMD). This was an observational retrospective clinical study that consecutively enrolled early BC patients with osteopenia to compare the impact of alendronate versus denosumab on secondary osteoporosis prevention and pain control. We identified two groups of patients treated with denosumab 60 mg by subcutaneous injection once every six months or alendronate 70 mg orally once a week. All the patients underwent a baseline physiatric evaluation (T0) and underwent a follow-up visit after 18 months (T1) together with femoral and vertebral Dual-Energy X-ray Absorptiometry (DEXA) exam evaluating T-Score marks. From September 2015 to December 2019 a total of 50 early (stage I-III) BC patients were considered eligible and consecutively enrolled in our study if they met pre-specified inclusion criteria. In the entire observed population, the addition of treatment with alendronate or denosumab led to a significant T-score improvement at the lumbar spine level (-1.92 vs -1.52, p=0.03), with a comparable contribution from alendronate (-1.60 vs -1.45, p=0.07) and denosumab (-2.26 vs -1.58, p=0.07). Regarding the femoral region, neither alendronate (-0.98 vs -1.07, p=0.23) nor denosumab (-1.39 vs -1.34, p=0.81) were able to produce any statistically relevant effect. However, concerning pain control, BMAs had a significant impact on reducing NRS scoresin the general population (T1 3.94 vs. baseline 4.32, p=0.007), with a likelyspecific contribution from alendronate (T1 3.52 vs. baseline 3.88, p=0.004) compared to denosumab (T1 4.36 vs baseline 4.76, p=0.12), without any differences in analgesic therapy assumption over time (p=0.93). Both alendronate and denosumab significantly contributed to preventing secondary osteoporosis in early BC patients with low BMD undergoing AIs, mostly at the lumbar spine level. Moreover, alendronate seemed to significantly impact pain control in such patients further supporting alendronate as a cost-effective option in this frail setting, although BMAs particularities should be carefully considered on an individual basis according to specific clinical contexts.


Phylogenetic diversity of Sri Lankan freshwater crabs and its implications for conservation.
As part of a Global Biodiversity Hotspot, the conservation of Sri Lanka's endemic biodiversity warrants special attention. With 51 species (50 of them endemic) occurring in the island, the biodiversity of freshwater crabs is unusually high for such a small area (65,600 km(2)). Freshwater crabs have successfully colonized most moist habitats and all climatic and elevational zones in Sri Lanka. We assessed the biodiversity of these crabs in relation to the different elevational zones (lowland, upland and highland) based on both species richness and phylogenetic diversity. Three different lineages appear to have radiated simultaneously, each within a specific elevational zone, with little interchange thereafter. The lowland and upland zones show a higher species richness than the highland zone while--unexpectedly--phylogenetic diversity is highest in the lowland zone, illustrating the importance of considering both these measures in conservation planning. The diversity indices for the species in the various IUCN Red List categories in each of the three zones suggest that risk of extinction may be related to elevational zone. Our results also show that overall more than 50% of Sri Lanka's freshwater crab species (including several as yet undescribed ones), or approximately 72 million years of evolutionary history, are threatened with extinction.


The experiences of carers in supporting a person living with dementia to participate in virtual cognitive stimulation therapy during the COVID-19 pandemic.
Cognitive stimulation therapy (CST) is an evidence-based group intervention for people with dementia. In response to the COVID-19 pandemic, many existing CST groups moved virtually and this required carers of people living with dementia to assist with setting up the appropriate technology. This study aimed to explore the roles and experiences of carers in accessing virtual CST (vCST). This qualitative study used semi-structured individual interviews, via telephone or videoconference, to explore the roles and experiences of carers. Interviews were recorded, transcribed verbatim and thematically analysed. Twelve family carers (age: 51-75 years) reported a range of experiences, from novice to expert knowledge in terms of accessing digital devices (mainly laptops and iPads) and connecting to Zoom. Accessing vCST provided carers an immediate application of new knowledge. Carers reported positive responses to vCST that provided their family member living with dementia with social contact and cognitive stimulation during lockdown. Accessing vCST required carers to have an existing adequate level of technological competence in order to learn and use the Zoom platform. Adult learning principles can be used to improve carers' digital literacy required for vCST and other telehealth initiatives.


Clinical laboratory urine analysis: comparison of the UriSed automated microscopic analyzer and the manual microscopy.
Several automated urine sediment analyzers have been introduced to clinical laboratories. Automated microscopic pattern recognition is a new technique for urine particle analysis. We evaluated the analytical and diagnostic performance of the UriSed automated microscopic analyzer and compared with manual microscopy for urine sediment analysis. Precision, linearity, carry-over, and method comparison were carried out. A total of 600 urine samples sent for urinalysis were assessed using the UriSed automated microscopic analyzer and manual microscopy. Within-run and between-run precision of the UriSed for red blood cells (RBC) and white blood cells (WBC) were acceptable at all levels (CV < 20%). Within-run and between-run imprecision of the UriSed testing for cast, squamous epithelial cells (EPI), and bacteria (BAC) were good at middle level and high level (CV < 20%). The linearity analysis revealed substantial agreement between the measured value and the theoretical value of the UriSed for RBC, WBC, cast, EPI, and BAC (r > 0.95). There was no carry-over. RBC, WBC, and squamous epithelial cells with sensitivities and specificities were more than 80% in this study. There is substantial agreement between the UriSed automated microscopic analyzer and the manual microscopy methods. The UriSed provides for a rapid turnaround time.


Latency and reactivation of JC virus in peripheral blood of human immunodeficiency virus type 1-infected patients.
JC virus (JCV) acts as an opportunistic virus in immunocompromised human immunodeficiency virus type 1 (HIV-1)-infected patients. The role of peripheral blood cells in central nervous system invasion, before the onset of progressive multifocal leukoencephalopathy (PML), remains controversial. In order to clarify JCV latency or reactivation status in peripheral blood, 72 HIV-1-infected patients were studied, together with 7 HIV-1-positive PML patients and 50 blood donors. Blood leukocytes, plasma, and B lymphocytes were investigated by two complementary DNA amplification procedures within the early T and late VP1 JCV genes and two reverse transcription techniques for the detection of corresponding early transcripts and mRNAs. JCV DNA was detected in 40.3% of the HIV-1-infected patients but only 8% of the blood donors (P < 0.001). Leukocytes represented 82.7% of the positive samples, but plasma from 12 patients (41.4%) contained JCV DNA. B lymphocytes seemed to be involved in the natural history of JCV but did not represent the unique cell target. JCV DNA was intermittently found in blood, and JCV mRNAs for VP1 capsid protein were detected exclusively in one PML patient. Such observations demonstrate that JCV, when detected in blood, does not undergo active multiplication. They support the JCV hematogenous spread hypothesis, but do not indicate any direct link between peripheral virus and dissemination in the central nervous system at the time of immunodepression.


Clinical research on antiangiogenic therapy.
A promising novel target for anti-cancer therapy is the tumour vasculature. Both primary tumour growth and the formation of metastasis depend on the establishment of new blood vessels from preexisting ones, a process called angiogenesis. Inhibition of angiogenesis and targeting of the tumour vasculature are highly effective in controlling tumour growth. Several natural and synthetic compounds have been developed and are currently tested in pre-clinical and clinical studies. This review intends to provide an overview of agents with antiangiogenic activity and the early clinical experience with these drugs.


Construction and characterization of deletion mutants of pseudorabies virus: a new generation of 'live' vaccines.
Various deletions were introduced into a cloned subgenomic fragment (BamHI-7), located in the unique short (US) region of the DNA from the virulent Northern Ireland Aujeszky-3 (NIA-3) strain of pseudorabies virus (PRV). In the cloned HindIII-B fragment, the MluI-BglII fragment was replaced by different MluI-BglII fragments of the deleted BamHI-7 clones. Transfection of the deleted HindIII-B fragments together with the HindIII-A fragment of either the NIA-3 or the non-virulent NIA-4 strain yielded replication-competent deletion mutants. The region in US in which sequences were deleted specified several mRNAs. Some of the mRNAs present in cells infected with NIA-3 were absent from cells infected with the deletion mutants, whereas other differently sized mRNAs were generated. The mutants were examined with respect to their biological properties in cell culture, mice and pigs. The results showed that the type of cytopathic effect induced in cell culture seemed to be determined by the UL region, using the mean time to death in mice as a parameter, markers for virulence were present in the US and UL regions and the introduction of deletions in US strongly reduced the virulence of PRV for pigs. Despite the impaired capacity of the deletion mutants to induce high titres of neutralizing antibodies in the serum, inoculation with mutants derived from NIA-3 prevented clinical disease in pigs upon challenge with the virulent parent strain. These deletion mutants provide a good basis for the production of bioengineered live PRV vaccines.


A patient-derived orthotopic xenograft (PDOX) mouse model of a cisplatinum-resistant osteosarcoma lung metastasis that was sensitive to temozolomide and trabectedin: implications for precision oncology.
In the present study, we evaluated the efficacy of trabectedin (TRAB) and temozolomide (TEM) compared to cisplatinum (CDDP) on a patient-derived orthotopic xenogrraft (PDOX) of a lung-metastasis from an osteosarcoma of a patient who failed CDDP therapy. Osteosarcoma resected from the patient was implanted orthotopically in the distal femur of mice to establish PDOX models which were randomized into the following groups when tumor volume reached approximately 100 mm3: G1, control without treatment; G2, CDDP (6 mg/kg, intraperitoneal injection, weekly, for 2 weeks); G3, TRAB (0.15 mg/kg, intravenous injection, weekly, for 2 weeks); G4, TEM (25 mg/kg, oral, daily, for 14 days). Tumor size and body weight were measured with calipers and a digital balance, respectively, twice a week. On day 14 after initiation of treatment, TEM and TRAB, but not CDDP, significantly inhibited tumor volume compared to untreated control: control (G1): 814.5±258.8 mm3; CDDP (G2): 608.6±126.9 mm3; TRAB (G3): 286.6±133.0 mm3; TEM (G4): 182.9±69.1 mm3. CDDP vs. control, p=0.07; TRAB vs. control, p=0.0004; TEM vs. control p =0.0002; TRAB vs. CDDP, p =0.0002; TEM vs. CDDP, p =0.00003. The results of the present study show that a PDOX model of an osteosarcoma lung-metastasis that recurred after adjuvant CDDP-treatment has identified potentially, highly-effective drugs for this recalcitrant disease, while accurately maintaining the CDDP resistance of the tumor in the patient, thereby demonstrating the potential of the osteosarcoma PDOX model for precision oncology.


Comparative phylogeography in rainforest trees from Lower Guinea, Africa.
Comparative phylogeography is an effective approach to assess the evolutionary history of biological communities. We used comparative phylogeography in fourteen tree taxa from Lower Guinea (Atlantic Equatorial Africa) to test for congruence with two simple evolutionary scenarios based on physio-climatic features 1) the W-E environmental gradient and 2) the N-S seasonal inversion, which determine climatic and seasonality differences in the region. We sequenced the trnC-ycf6 plastid DNA region using a dual sampling strategy: fourteen taxa with small sample sizes (dataset 1, mean n = 16/taxon), to assess whether a strong general pattern of allele endemism and genetic differentiation emerged; and four taxonomically well-studied species with larger sample sizes (dataset 2, mean n = 109/species) to detect the presence of particular shared phylogeographic patterns. When grouping the samples into two alternative sets of two populations, W and E, vs. N and S, neither dataset exhibited a strong pattern of allelic endemism, suggesting that none of the considered regions consistently harboured older populations. Differentiation in dataset 1 was similarly strong between W and E as between N and S, with 3-5 significant F ST tests out of 14 tests in each scenario. Coalescent simulations indicated that, given the power of the data, this result probably reflects idiosyncratic histories of the taxa, or a weak common differentiation pattern (possibly with population substructure) undetectable across taxa in dataset 1. Dataset 2 identified a common genetic break separating the northern and southern populations of Greenwayodendron suaveolens subsp. suaveolens var. suaveolens, Milicia excelsa, Symphonia globulifera and Trichoscypha acuminata in Lower Guinea, in agreement with differentiation across the N-S seasonal inversion. Our work suggests that currently recognized tree taxa or suspected species complexes can contain strongly differentiated genetic lineages, which could lead to misinterpretation of phylogeographic patterns. Therefore the evolutionary processes of such taxa require further study in African tropical rainforests.


A draft framework for measuring progress towards the development of a National Health Information Infrastructure.
American public policy makers recently established the goal of providing the majority of Americans with electronic health records by 2014. This will require a National Health Information Infrastructure (NHII) that is far more complete than the one that is currently in its formative stage of development. We describe a conceptual framework to help measure progress toward that goal. The NHII comprises a set of clusters, such as Regional Health Information Organizations (RHIOs), which, in turn, are composed of smaller clusters and nodes such as private physician practices, individual hospitals, and large academic medical centers. We assess progress in terms of the availability and use of information and communications technology and the resulting effectiveness of these implementations. These three attributes can be studied in a phased approach because the system must be available before it can be used, and it must be used to have an effect. As the NHII expands, it can become a tool for evaluating itself. The NHII has the potential to transform health care in America--improving health care quality, reducing health care costs, preventing medical errors, improving administrative efficiencies, reducing paperwork, and increasing access to affordable health care. While the President has set an ambitious goal of assuring that most Americans have electronic health records within the next 10 years, a significant question remains "How will we know if we are making progress toward that goal?" Using the definitions for "nodes" and "clusters" developed in this article along with the resulting measurement framework, we believe that we can begin a discussion that will enable us to define and then begin making the kinds of measurements necessary to answer this important question.


Determination of Genotypic and Phenotypic Characteristics of Friedreich's Ataxia and Autosomal Dominant Spinocerebellar Ataxia Types 1, 2, 3, and 6.
This study aimed to analyze the genotypic characteristics of Friedreich's ataxia (FA) and autosomal dominant ataxias [such as spinocerebellar ataxia (SCA) types 1, 2, 3, and 6] using molecular and biological methods in hereditary cerebellar ataxia considering both clinical and electrophysiological findings. The study included 129 indexed cases, who applied to the neurology department and were diagnosed with hereditary cerebellar ataxia through clinical, laboratory, and electrophysiological findings, and 15 sibling patients who were diagnosed through family scanning (144 cases in total); their genetic analyses were also performed. Detailed physical and neurological examinations, pedigree analyses, electroneurography, evoked potentials, cerebral-spinal magnetic resonance imaging, and echocardiographic analyses were performed for all cases. Blood samples were collected from patients, and the genotypic characteristics of autosomal dominant SCA types 1, 2, 3, and 6 were investigated. Statistical analyses were performed with the Statistical Package for the Social Sciences (SPSS Inc; Chicago, IL, USA) 17.0. Almost 50% of patients were defined as FA. Moreover, two SCA1 cases and one SCA6 case were detected. In our study, 47.2% of patients with FA had developed hereditary cerebellar ataxia. Ground and autosomal dominant-linked SCA1 and SCA6 were each detected in one family. These data suggest that patients with cerebellar ataxia of hereditary origin should be primarily examined for FA.


Symbolic Learning and Reasoning With Noisy Data for Probabilistic Anchoring.
Robotic agents should be able to learn from sub-symbolic sensor data and, at the same time, be able to reason about objects and communicate with humans on a symbolic level. This raises the question of how to overcome the gap between symbolic and sub-symbolic artificial intelligence. We propose a semantic world modeling approach based on bottom-up object anchoring using an object-centered representation of the world. Perceptual anchoring processes continuous perceptual sensor data and maintains a correspondence to a symbolic representation. We extend the definitions of anchoring to handle multi-modal probability distributions and we couple the resulting symbol anchoring system to a probabilistic logic reasoner for performing inference. Furthermore, we use statistical relational learning to enable the anchoring framework to learn symbolic knowledge in the form of a set of probabilistic logic rules of the world from noisy and sub-symbolic sensor input. The resulting framework, which combines perceptual anchoring and statistical relational learning, is able to maintain a semantic world model of all the objects that have been perceived over time, while still exploiting the expressiveness of logical rules to reason about the state of objects which are not directly observed through sensory input data. To validate our approach we demonstrate, on the one hand, the ability of our system to perform probabilistic reasoning over multi-modal probability distributions, and on the other hand, the learning of probabilistic logical rules from anchored objects produced by perceptual observations. The learned logical rules are, subsequently, used to assess our proposed probabilistic anchoring procedure. We demonstrate our system in a setting involving object interactions where object occlusions arise and where probabilistic inference is needed to correctly anchor objects.


Tryptophan and arginine catabolic enzymes and regulatory cytokines in clinically isolated syndrome and multiple sclerosis.
Clinically isolated syndrome (CIS) is the earliest clinical episode in multiple sclerosis (MS). A study of circulating cells from patients with CIS may help us understand the transition to, and processes associated with, the development of MS. As immune cell activity can be determined by flux through metabolic pathways, the mRNA expression of l-tryptophan- and l-arginine-catabolising enzymes, indoleamine 2,3-dioxygenase (IDO) 1 and IDO2 and arginase (ARG) 1 and ARG2, respectively, was compared between peripheral blood mononuclear cells (PBMCs) from healthy controls, and patients with CIS and definite MS. As one measure of cell function, cytokine mRNA levels were analysed directly ex vivo and in cells after culture for 4 h in the absence of regulatory factors in autologous serum. When measured directly ex vivo, the expression of IDO and ARG was greater in cells from patients with CIS and MS than cells from healthy controls. Although not linked to IDO and ARG expression, PBMCs from the CIS patients were characterised by low IL-10 and TGFB mRNA levels and not by greater expression of proinflammatory cytokines. When the cells were cultured for 4 h without autologous serum, pro- and anti-inflammatory cytokine mRNA levels positively correlated with IDO1 expression, and TGFB mRNA levels correlated with ARG1 expression. Higher IDO and ARG expression in CIS and MS provides one sustained homeostatic mechanism to control MS-associated inflammation. However, potent extrinsic mediators in serum may regulate immune cell function in CIS and associations between IDO, ARG and cytokine expression.


Mannose receptor is an HIV restriction factor counteracted by Vpr in macrophages.
HIV-1 Vpr is necessary for maximal HIV infection and spread in macrophages. Evolutionary conservation of Vpr suggests an important yet poorly understood role for macrophages in HIV pathogenesis. Vpr counteracts a previously unknown macrophage-specific restriction factor that targets and reduces the expression of HIV Env. Here, we report that the macrophage mannose receptor (MR), is a restriction factor targeting Env in primary human monocyte-derived macrophages. Vpr acts synergistically with HIV Nef to target distinct stages of the MR biosynthetic pathway and dramatically reduce MR expression. Silencing MR or deleting mannose residues on Env rescues Env expression in HIV-1-infected macrophages lacking Vpr. However, we also show that disrupting interactions between Env and MR reduces initial infection of macrophages by cell-free virus. Together these results reveal a Vpr-Nef-Env axis that hijacks a host mannose-MR response system to facilitate infection while evading MR's normal role, which is to trap and destroy mannose-expressing pathogens.


Serotonin Syndrome Complicating Treatment of Ifosfamide Neurotoxicity With Methylene Blue.
Methylene blue is a widely used treatment for ifosfamide neurotoxicity. We present a case of severe encephalopathy complicating ifosfamide-based therapy for recurrent retroperitoneal leiomyosarcoma. After treatment with methylene blue, the patient experienced clinical decompensation and was diagnosed with serotonin syndrome based on a constellation of clinical findings. Withdrawal of methylene blue and other serotonergic medications led to clinical stabilization and ultimately neurological recovery. Our case highlights the challenge of diagnosing serotonin syndrome in the face of preexisting ifosfamide neurotoxicity, as there is significant clinical overlap between these 2 syndromes. Practitioners must remain vigilant of this potential life-threatening complication in this vulnerable population.


[Spontaneous intracranial hypotension: a rare cause of chronic headache].
Spontaneous intracranial hypotension is a rare cause of chronic postural headache. We report the case of a 58-year-old woman with a 1.5 year history of chronic headache in upright position, nausea and vomiting. Neurological examination was normal. The Gd-enhanced MRI of the brain showed an abnormal meningeal enhancement over the cerebral convexity. The CSF was normal. Opening pressure during lumbar puncture was 6 cm H2O in the lateral recumbent position. No evidence of underlying systemic, infectious or neoplastic diseases could be detected. The headache was alleviated under theophylline therapy. Spontaneous intracranial hypotension should be considered in the differential diagnosis of chronic postural headache. MRI revealing meningeal enhancement and low opening pressure in lumbar puncture are important findings for the diagnosis of spontaneous intracranial hypotension.


Alpha,2-, alpha,3-, and alpha,4-dehydrophenol radical anions: formation, reactivity, and energetics leading to the heats of formation of alpha,2-, alpha,3-, and alpha,4-oxocyclohexadienylidene.
We have regiospecifically generated the alpha,2-, alpha,3-, and alpha,4-dehydrophenoxide anions by collisional activation of o-, m-, and p-nitrobenzoate. The alpha,2 and alpha,4 isomers also were synthesized by reacting o-benzyne radical anion with carbon dioxide and electron ionization of p-diazophenol. All three dehydrophenol radical anions were differentiated from each other and identified by probing their chemical reactivity with several reagents. Each isomer was converted to phenoxide and its corresponding quinone as well. Thermochemical measurements were carried out on all three radical anions and their hydrogen-atom affinities, proton affinities, and electron binding energies are reported. These measured quantities are combined in thermodynamic cycles to derive the heats of formation of each of the radical anions and their corresponding carbenes (i.e., alpha,2-, alpha,3-, and alpha,4-dehydrophenol). These results are compared to MCQDPT2, G3, G2+(MP2), and B3LYP calculations and experimental data for appropriate reference compounds.


Comparison of anesthetic management and outcomes of robot-assisted versus open radical cystectomy.
Robot-assisted cystectomy surgery may be advantageous for patients. The purpose of this study was to compare anesthetic management and outcomes in patients undergoing robot-assisted versus open radical cystectomy. In a retrospective review of 256 cystectomy procedures, procedure duration, blood loss, respiratory parameters, recovery room opiate consumption, pain scores and antiemetic use in the recovery room, and hospital length of stay were compared. After exclusions, 96 robot-assisted and 102 open procedures were analyzed. Anesthesia and surgery duration were significantly longer in the robot-assisted group, while the length of hospital stay was significantly shorter in the robot-assisted group: 7.1 ± 5.8 versus 9.8 ± 5.03 days, p = 0.0005. Estimated blood loss was 601.8 ± 491.4 ml in the open group versus 257.7 ± 164.3 ml in the robot-assisted group, p < 0.0001. Recovery room opiate consumption was significantly less in the robot-assisted group: 9.5 ± 8.9 versus 12.6 ± 9.9 mg (morphine equivalents), p = 0.02. The highest recorded respiratory rate was significantly higher in the robot-assisted group, as was the highest recorded peak airway pressure. Among patients with arterial blood gas data, the highest arterial partial pressure of CO2 was significantly greater in the robot-assisted group than in the open surgery group: 42.6 ± 5.6 versus 37.4 ± 4.8 mmHg CO2, p = 0.0001. Surgeons and anesthesia providers can expect robot-assisted radical cystectomy surgery to last longer than traditional open surgery, but to be associated with less pain and blood loss. Positioning and abdominal insufflation for robot-assisted surgery may contribute to ventilation challenges.


[The etiological structure of acute intestinal infections].
The share of various etiologic forms of acute intestinal infections, diagnosed by bacteriologic methods, is presented. The share of gastroenterocolitis induced by opportunistic microflora makes up 35.6%, that of dysentery 25.6%, salmonellosis 18.5%; mixed infection (dysentery + salmonellosis) is diagnosed in 7% of cases with acute intestinal infections. The principal representative of opportunistic microflora isolated from patients with acute intestinal infections is the Klebsiella genus (40.6%), whereas in the reference group Citrobacter, Morganella, and Klebsiella detection rates are approximately the same (27.0-18.3%). Opportunistic microorganisms in titers under 10(6) are isolated from normal subjects 5 times more frequently than from the patients, this indicating the diagnostic value of this level of feces contamination with opportunistic microflora.


A critical review of studies on vitamin D in relation to colorectal cancer.
Vitamin D intake has been hypothesized to reduce the risk of several types of cancer. Vitamin D and its analogues have demonstrated anticancer activity in vitro and in animal models. However, the risk of colorectal cancer in relation to dietary vitamin D remains controversial. A literature search was performed for articles on epidemiologic studies of vitamin D and colorectal cancer and the mechanisms involved. Studies that combine multiple sources of vitamin D or examine serum 25(OH)D3 usually find that above-average vitamin D intake and serum metabolite concentrations are associated with significantly reduced incidence of colorectal cancer. A number of mechanisms have been identified through which vitamin D may reduce the risk of colorectal and several other types of cancer. Although studies that include vitamin D from all sources or serum 25(OH)D3 usually show significantly reduced incidence of colorectal cancer in association with vitamin D, analyses limited to dietary vitamin D tend to have mixed results. The likely reason that dietary vitamin D is not a significant risk reduction factor for colorectal cancer in many studies is that dietary sources provide only a portion of total vitamin D, with supplements and synthesis of vitamin D in the skin in association with solar UV-B radiation providing the balance. There is strong evidence from several different lines of investigation supporting the hypothesis that vitamin D may reduce the risk of colorectal cancer. Further study is required to elucidate the mechanisms and develop guidelines for optimal vitamin D sources and serum levels of vitamin D metabolites.


Plasticity and proprioception in insects. II. Modes of reflex action of the locust metathoracic femoral chordotonal organ.
Reflex responses of tibial motoneurones were examined during mechanical stimulation of the femoral chordotonal organ, a joint angle receptor of the locust hindleg. Step displacements of the main ligament of the organ, mimicking 10-15 degree changes in joint angle, produced different patterns of discharge in motoneurones (1) when the leg was resting against a support and (2) when the support was removed to induce active searching movements. Tibial motoneurones showed resistance reflex responses to oppose the apparent joint movement when the leg rested against a support. Resistance reflexes consisted of constant, short latency excitatory responses followed by discharges that varied in intensity (gain) and degree of tonic coupling. These variations were not due to simple summation with other inputs to motoneurones. Responses changed during periods of active searching movements. Tibial flexor motoneurones fired phasically in response to apparent joint movement in any direction. Tibial extensor motoneurones were generally inhibited by chordotonal inputs. These reflex changes are not simple reflex 'reversals', but represent more complex changes in reflex mode. Potential functions of each of these reflex modes and the need for plasticity in reflexes of the chordotonal organ are discussed.


Enterovirus 71 3C inhibits cytokine expression through cleavage of the TAK1/TAB1/TAB2/TAB3 complex.
Enterovirus 71 (EV71) causes hand, foot, and mouth disease in young children and infants. Severe infection with EV71 can lead to various neurological complications or fatal diseases. However, the mechanism of EV71 pathogenesis is poorly understood. Emerging evidence suggests that EV71 modulates type I interferon (IFN) and cytokine responses. Here, we show that EV71 disables components of the TAB2 complex through the 3C protein. When expressed in mammalian cells, EV71 3C interacts with TAB2 and TAK1, which inhibits NF-κB activation. Furthermore, 3C mediates cleavage of TAB2 and its partners, which requires the protease activity. H40D or C147S substitution in the 3C active sites abolishes its activity, whereas R84Q or V154S substitution in the RNA binding domain has no effect. The 3C protein targets TAB2 at Q113-S114, TAK1 at Q360-S361, TAB1 both at Q414-G415 and Q451-S452, and TAB3 at Q173-G174 and Q343-G344. Importantly, overexpression of TAB2 inhibits EV71 replication, whereas addition of cleaved fragments has no effect. Thus, an equilibrium between the TAB2 complex and EV71 3C represents a control point of viral infection. These results suggest that TAK1/TAB1/TAB2/TAB3 cleavage mediated by EV71 may be a mechanism to interfere with inflammatory responses. The TAK1 complex plays a critical role in the activation of NF-κB and cytokine production. However, little is known about its connection to enterovirus 71 (EV71). We demonstrate that EV71 3C suppresses cytokine expression via cleavage of the TAK1 complex proteins. EV71 3C interacts with TAB2 and TAK1. Furthermore, overexpression of TAB2 inhibits EV71 replication, whereas addition of cleaved fragment has no effect. These results suggest that the interplay of EV71 and the TAK1 complex influences the outcome of viral infection.


"Sympathy" vs."Empathy": Comparing experiences of I2Audits and disability simulations.
People with disability often experience stigma and discrimination, and people with disability in rural areas may experience these at higher rates. Additionally, people with disability in rural areas may have fewer opportunities for physical and social participation due to barriers in the built environment. Activities such as disability simulations and inclusive, interdisciplinary community planning workshops (i.e., I2Audits) seek to draw awareness to and address these problematic experiences. The present study used thematic analysis from qualitative research to examine the advantages and disadvantages of using disability simulations and I2Audits in rural communities. Findings suggest that disability simulations increase stigmatization, lead to feelings of embarrassment and discomfort, and do not capture the experiences of people with disability. On the other hand, I2Audits lead to meaningful environmental changes, create feelings of empowerment, and center the lived experiences of people with disability within a bio-psycho-social model of disability. Results suggest that not only can I2Audits be a powerful tool to draw attention to physical barriers that people with disability face, but they also draw attention to the multi-level changes needed to increase opportunities for participation and address sources of stigma and discrimination in rural areas.


Structural and spectral response of Aequorea victoria green fluorescent proteins to chromophore fluorination.
Global replacements of tyrosine by 2- and 3-fluorotyrosine in "enhanced green" and "enhanced yellow" mutants of Aequorea victoria green fluorescent proteins (avGFPs) provided protein variants with novel biophysical properties. While crystallographic and modeled structures of these proteins are indistinguishable from those of their native counterparts (i.e., they are perfectly isomorphous), there are considerable differences in their spectroscopic properties. The fluorine being an integral part of the avGFP chromophore induces changes in the titration curves, variations in the intensity of the absorbance and fluorescence, and spectral shifts in the emission maxima. Furthermore, targeted fluorination in close proximity to the fluorinated chromophore yielded additional variants with considerably enhanced spectral changes. These unique spectral properties are intrinsic features of the fluorinated avGFPs, in the context of the rigid chromophore-microenvironment interactions. The availability of the isomorpohous crystal structures of fluorinated avGFPs allowed mapping of novel, unusual interaction distances created by the presence of fluorine atoms. In addition, fluorine atoms in the ortho position of the chromophore tyrosyl moiety exhibit a single conformation, while in the meta position two conformer states were observed in the crystalline state. Such global replacements in chromophores of avGFPs and similar proteins result in "atomic mutations" (i.e., H --> F replacements) in the structures, offering unprecedented opportunities to understand and manipulate the relationships between protein structure and spectroscopic properties.


Identification of a phosphoprotein in the nuclear matrix by monoclonal antibodies.
Previous work in this laboratory has established that a rat liver nuclear phosphoprotein (B2:Mr 68,000, pI 6.5-8.2) is associated with actively transcribed nucleosomes, as demonstrated by its preferential release after mild treatment with micrococcal nuclease. In the present report we provide further immunological evidence ('Western Blot' analysis, solid-phase radioimmunoassay and indirect immunofluorescence) that in addition establishes the presence of this phosphoprotein in the nuclear-matrix protein fraction. This paradoxical localization suggests that this phosphoprotein may function in two separate and distinct roles within the realm of nuclear organization.


Predictive factors for successful clinical outcome 1 year after an intensive combined physical and psychological programme for chronic low back pain.
The aim of this longitudinal study is to determine the factors which predict a successful 1-year outcome from an intensive combined physical and psychological (CPP) programme in chronic low back pain (CLBP) patients. A prospective cohort of 524 selected consecutive CLBP patients was followed. Potential predictive factors included demographic characteristics, disability, pain and cognitive behavioural factors as measured at pre-treatment assessment. The primary outcome measure was the oswestry disability index (ODI). A successful 1-year follow-up outcome was defined as a functional status equivalent to 'normal' and healthy populations (ODI ≤22). The 2-week residential programme fulfills the recommendations in international guidelines. For statistical analysis we divided the database into two equal samples. A random sample was used to develop a prediction model with multivariate logistic regression. The remaining cases were used to validate this model. The final predictive model suggested being 'in employment' at pre-treatment [OR 3.61 (95 % CI 1.80-7.26)] and an initial 'disability score' [OR 0.94 (95 % CI 0.92-0.97)] as significant predictive factors for a successful 1-year outcome (R (2) = 22 %; 67 % correctly classified). There was no predictive value from measures of psychological distress. CLBP patients who are in work and mild to moderately disabled at the start of a CPP programme are most likely to benefit from it and to have a successful treatment outcome. In these patients, the disability score falls to values seen in healthy populations. This small set of factors is easily identified, allowing selection for programme entry and triage to alternative treatment regimes.


"Gender-based Violence (GBV) and HIV, they are like sister and brother": barriers and facilitators to GBV screening and referral in public HIV treatment settings in Uganda.
People living with HIV are vulnerable to gender-based violence (GBV), which can negatively impact HIV treatment outcomes. National guidelines in Uganda recommend GBV screening alongside HIV treatment services. We explored barriers and facilitators to providers implementing GBV screening and referral in public antiretroviral therapy (ART) clinics in Uganda. We conducted qualitative in-depth interviews. Providers were purposively sampled from 12 ART clinics to represent variation in clinical specialty and gender. We used the Theoretical Domains Framework to structure our deductive analysis. We conducted 30 in-depth interviews with providers implementing GBV screening and/or referral. Respondents had a median age of 36 (IQR: 30, 43) years and had been offering post-GBV care to clients for a median duration of 5 (4, 7) years. 67% of respondents identified as female and 57% were counselors. Facilitators of GBV screening and referral included providers having access to post-GBV standard operating procedures and screening tools, trainings offered by the Ministry of Health, facility-sponsored continuing medical education units and support from colleagues. Respondents indicated that referrals were uncommon, citing the following barriers: negative expectations regarding the quality and quantity of referral services; lack of financial resources to support clients, facilities, and referral partners throughout the referral process; and sociocultural factors that threatened client willingness to pursue post-GBV support services. Findings from this evaluation support the refinement of GBV screening and referral implementation strategies that leverage facilitators and address barriers to better support individuals living with HIV and who may have heightened vulnerability to GBV.


Lubricin (PRG-4) anti-fouling coating for surface-enhanced Raman spectroscopy biosensing: towards a hierarchical separation system for analysis of biofluids.
Surface-enhanced Raman Spectroscopy (SERS) is a powerful optical sensing technique that amplifies the signal generated by Raman scattering by many orders of magnitude. Although the extreme sensitivity of SERS enables an extremely low limit of detection, even down to single molecule levels, it is also a primary limitation of the technique due to its tendency to equally amplify 'noise' generated by non-specifically adsorbed molecules at (or near) SERS-active interfaces. Eliminating interference noise is thus critically important to SERS biosensing and typically involves onerous extraction/purification/washing procedures and/or heavy dilution of biofluid samples. Consequently, direct analysis within biofluid samples or in vivo environments is practically impossible. In this study, an anti-fouling coating of recombinant human Lubricin (LUB) was self-assembled onto AuNP-modified glass slides via a simple drop-casting method. A series of Raman spectra were collected using rhodamine 6G (R6G) as a model analyte, which was spiked into NaCl solution or unprocessed whole blood. Likewise, we demonstrate the same sensing system for the quantitative detection of L-cysteine spiked in undiluted milk. It was demonstrated for the first time that LUB coating can mitigate the deleterious effect of fouling in a SERS sensor without compromising the detection of a target analyte, even in a highly fouling, complex medium like whole blood or milk. This feat is achieved through a molecular sieving property of LUB that separates small analytes from large fouling species directly at the sensing interface resulting in SERS spectra with low background (i.e., noise) levels and excellent analyte spectral fidelity. These findings indicate the great potential for using LUB coatings together with an analyte-selective layer to form a hierarchical separation system for SERS sensing of relevant analytes directly in complex biological media, aquaculture, food matrix or environmental samples.


Gene expression profile analysis of colorectal cancer to investigate potential mechanisms using bioinformatics.
This study aimed to explore the underlying molecular mechanisms of colorectal cancer (CRC) using bioinformatics analysis. Using GSE4107 datasets downloaded from the Gene Expression Omnibus, the differentially expressed genes (DEGs) were screened by comparing the RNA expression from the colonic mucosa between 12 CRC patients and ten healthy controls using a paired t-test. The Gene Ontology (GO) functional and pathway enrichment analyses of DEGs were performed using the Database for Annotation, Visualization and Integrated Discovery (DAVID) software followed by the construction of a protein-protein interaction (PPI) network. In addition, hub gene identification and GO functional and pathway enrichment analyses of the modules were performed. A total of 612 up- and 639 downregulated genes were identified. The upregulated DEGs were mainly involved in the regulation of cell growth, migration, and the MAPK signaling pathway. The downregulated DEGs were significantly associated with oxidative phosphorylation, Alzheimer's disease, and Parkinson's disease. Moreover, FOS, FN1, PPP1CC, and CYP2B6 were selected as hub genes in the PPI networks. Two modules (up-A and up-B) in the upregulated PPI network and three modules (d-A, d-B, and d-C) in the downregulated PPI were identified with the threshold of Molecular Complex Detection (MCODE) Molecular Complex Detection (MCODE) score ≥4 and nodes ≥6. The genes in module up-A were significantly enriched in neuroactive ligand-receptor interactions and the calcium signaling pathway. The genes in module d-A were enriched in four pathways, including oxidative phosphorylation and Parkinson's disease. DEGs, such as FOS, FN1, PPP1CC, and CYP2B6, may be used as potential targets for CRC diagnosis and treatment.


Vitamin C deficiency in the brain impairs cognition, increases amyloid accumulation and deposition, and oxidative stress in APP/PSEN1 and normally aging mice.
Subclinical vitamin C deficiency is widespread in many populations, but its role in both Alzheimer's disease and normal aging is understudied. In the present study, we decreased brain vitamin C in the APPSWE/PSEN1deltaE9 mouse model of Alzheimer's disease by crossing APP/PSEN1(+) bigenic mice with SVCT2(+/-) heterozygous knockout mice, which have lower numbers of the sodium-dependent vitamin C transporter required for neuronal vitamin C transport. SVCT2(+/-) mice performed less well on the rotarod task at both 5 and 12 months of age compared to littermates. SVCT2(+/-) and APP/PSEN1(+) mice and the combination genotype SVCT2(+/-)APP/PSEN1(+) were also impaired on multiple tests of cognitive ability (olfactory memory task, Y-maze alternation, conditioned fear, Morris water maze). In younger mice, both low vitamin C (SVCT2(+/-)) and APP/PSEN1 mutations increased brain cortex oxidative stress (malondialdehyde, protein carbonyls, F2-isoprostanes) and decreased total glutathione compared to wild-type controls. SVCT2(+/-) mice also had increased amounts of both soluble and insoluble Aβ1-42 and a higher Aβ1-42/1-40 ratio. By 14 months of age, oxidative stress levels were similar among groups, but there were more amyloid-β plaque deposits in both hippocampus and cortex of SVCT2(+/-)APP/PSEN1(+) mice compared to APP/PSEN1(+) mice with normal brain vitamin C. These data suggest that even moderate intracellular vitamin C deficiency plays an important role in accelerating amyloid pathogenesis, particularly during early stages of disease development, and that these effects are likely modulated by oxidative stress pathways.


The Nightmare Disorder Index: development and initial validation in a sample of nurses.
Nurses are a group at high risk for nightmares, yet little is known about the rate of nightmare disorder and associated psychosocial factors in this group in part attributable to the lack of a self-report questionnaire to assess DSM-5 criteria for nightmare disorder. Aims of the current study were to (1) report on development and initial validity of a self-report measure of DSM-5 nightmare disorder, and (2) examine the rate and associated factors of nightmare disorder among nurses. Nurses (N = 460) completed baseline measures online including Nightmare Disorder Index (NDI), psychosocial and demographic questionnaires. A subset (n = 400) completed 14 days of sleep diaries and actigraphy. NDI demonstrated satisfactory psychometric characteristics as indicated by good internal consistency (α = 0.80), medium inter-item correlations (r = 0.50), medium to large item-total (r = 0.55-0.85) and convergent correlations (0.32-0.45), and small to medium discriminant correlations (-0.12-0.33). Per NDI, 48.7% of nurses reported no nightmares in the past month, 43.9% met partial/subthreshold criteria and 7.4% met full criteria for probable nightmare disorder. Nurses with nightmare disorder demonstrated significantly poorer psychosocial functioning (i.e. posttraumatic stress, depression, anxiety, stress) than those with subthreshold nightmare symptoms, who had poorer functioning than those with no nightmares. NDI is an efficient and valid self-report assessment of nightmare disorder. Nurses have high rates of nightmares and nightmare disorder which are associated with poorer psychosocial functioning. We recommend increased nightmare screening particularly for high-risk populations such as healthcare workers.


Warning about warnings: weighing risk and benefit when information is in a state of flux.
In 2010, new data about the safety of recombinant human growth hormone (rhGH) resulted in warnings and subsequent pronouncements by the US Food and Drug Administration (FDA) and its European counterpart [the European Medicines Agency (EMA)] regarding its use in children and adolescents as an elective treatment for short stature. However, opinions about these new data are divergent: the FDA did not change the label of the drug and experts have argued for further research on the safety of rhGH. In this situation of an evolving scientific controversy, it is unclear how questions about benefit and risk are communicated to patients and their parents. Social biases and misperceptions about the deleterious effects of short stature and the benefits of added height influence decisions to prescribe rhGH and may affect discussions of the warnings by regulators. Fully supporting a model of shared decision-making involving children and adolescents requires sharing risk-benefit information, including evolving information from drug regulators, with patients and parents.


Conventional measurement of renal function utilizing serum creatinine, creatinine clearance, inulin and para-aminohippuric acid clearance.
Inulin and para-aminohippuric acid clearances, determined by the conventional method of continuous intravenous infusion with blood and urine sample collections, are the gold standards for estimating glomerular filtration rate and renal plasma flow, respectively. Creatinine clearance provides a reasonably good estimate of glomerular filtration rate but is still subject to errors in accuracy and precision. However, novel methods employing cimetidine to block renal tubular creatinine secretion hold promise for improving the accuracy of estimates. More importantly, a large (and growing) number of studies have consistently demonstrated that estimating glomerular filtration rate by creatinine clearance calculated from the Cockcroft-Gault formula is better than measuring creatinine clearance with a 24-h urine collection. Until newer, more simple methods are developed, calculating creatinine clearance using fasting serum creatinine level, body weight, age and sex provides a reasonable and clinically useful bedside measure of glomerular filtration rate for the practising clinician.


Collie Eye Anomaly in Switzerland.
In this retrospective study, the results of 3'527 eye examinations in 6 different breeds affected with Collie Eye Anomaly (CEA) over a period of 8 years (1999 - 2007) are described. CEA was divided into three main ophthalmoscopic features, a) choroidal hypoplasia (CRH), b) CRH and coloboma and c) coloboma alone. Of the 101 Smooth Collies 8.9 % showed signs of CRH, whereas 36.9 % of Rough Collies were affected with CRH, 2.8 % with CRH and coloboma and 0.38 % with coloboma alone. Choroidal hypoplasia was present in 13.1 %, CRH and coloboma in 1.8 % and coloboma alone in 0.2 % of the Shetland Sheepdogs. Only one Australian Shepherd dog had CRH, while 0.7 % of the Border Collies were affected with CRH. None of the Nova Scotia Duck Tolling Retrievers were affected with CEA. There were no statistically significant differences in the occurrence of CEA between males and females, nor was there any relation between coat colors. Significant differences could be shown between dogs younger or older than 8 weeks at first examination. CEA was more often diagnosed in dogs younger than 8 weeks within the Rough Collie and Shetland Sheepdog.


Calreticulin: one protein, one gene, many functions.
The endoplasmic reticulum (ER) plays a critical role in the synthesis and chaperoning of membrane-associated and secreted proteins. The membrane is also an important site of Ca(2+) storage and release. Calreticulin is a unique ER luminal resident protein. The protein affects many cellular functions, both in the ER lumen and outside of the ER environment. In the ER lumen, calreticulin performs two major functions: chaperoning and regulation of Ca(2+) homoeostasis. Calreticulin is a highly versatile lectin-like chaperone, and it participates during the synthesis of a variety of molecules, including ion channels, surface receptors, integrins and transporters. The protein also affects intracellular Ca(2+) homoeostasis by modulation of ER Ca(2+) storage and transport. Studies on the cell biology of calreticulin revealed that the ER membrane is a very dynamic intracellular compartment affecting many aspects of cell physiology.


Inhibition of Ca absorptive flux by chlorthalidone in the rat duodenum and colon.
To evaluate the mode and intestinal sites of action of chlorthalidone on Ca transport, balance and flux studies by the Ussing technique were performed in rats fed a low Ca diet throughout the experiment to stimulate Ca absorption. Half of the rats received chlorthalidone (5 mg X kg-1 X day-1) orally for 18 wk, a duration of administration previously shown to yield consistent changes. Net Ca absorption was decreased by chlorthalidone (0.91 +/- 0.07 vs. 1.08 +/- 0.3 mg/day, P less than 0.05), which is similar to prior observations in rats fed a normal Ca diet. Ca balance was unchanged by chlorthalidone (0.34 +/- 0.08 vs. 0.36 +/- 0.05 mg/day) because of concomitant hypocalciuria of similar magnitudes. In the duodenum, chlorthalidone decreased mucosal-to-serosal flux (Jm----s) (96.7 vs. 111.5 nmol X cm-2 X h-1, P less than 0.05). Since serosal-to-mucosal flux (Js----m) was unchanged, net Ca flux (Jnet) was reduced (74.0 vs. 89.6 nmol X cm-2 X h-1, P less than 0.05). Similarly, chlorthalidone selectively inhibited Jm----s in the descending colon (27.0 vs. 36.2 nmol X cm-2 X h-1, P less than 0.01), reducing Jnet (14.3 vs. 23.2 nmol X cm-2 X h-1, P less than 0.005). Short-circuit current and tissue conductance were unaffected. These data suggest that chronic chlorthalidone treatment reduces Ca absorption by inhibiting active calcium transport in the small and large intestine.


Influenza C virus CM2 protein is produced from a 374-amino-acid protein (P42) by signal peptidase cleavage.
Although unspliced mRNA from influenza C virus RNA segment 6 (M gene) has a single open reading frame capable of encoding a 374-amino-acid protein (Mr, 42,000), the major polypeptide synthesized from this mRNA species is the CM2 protein, with an Mr of 18,000. The present study was performed to investigate the mechanism by which CM2 is generated from the unspliced mRNA. It was reported previously that the 374-amino-acid protein (P42) is an integral membrane protein having two internal hydrophobic domains, one of which (residues 241 to 252) is followed by two sequences (252 Ile-Thr-Ser and 257 Ala-Ser-Ala) favorable for cleavage by signal peptidase. To examine the possibility that P42 is cleaved by signal peptidase after Ser residue 254 or Ala residue 259 to yield CM2, we constructed three mutated M gene cDNAs in which either or both of the two sequences were eliminated and tested their ability to synthesize CM2 in the transfected COS cells. The results showed that CM2 synthesis was blocked completely when the second recognition motif for signal peptidase was removed. It was also found that when the mRNA transcript of the wild-type M gene was translated in vitro, P42, but not CM2, was synthesized in the absence of dog pancreas microsomal membranes, whereas CM2, in addition to a polypeptide (designated M1') slightly larger than matrix protein (M1), was synthesized in the presence of microsomes. When the same experiment was done with the transcript of the mutated M gene in which the second recognition motif was removed, synthesis of CM2 could not be seen, even in the presence of microsomes. From these results, we conclude that cleavage of P42 by signal peptidase after Ala residue 259 produces CM2, composed of the C-terminal 115 amino acids, in addition to M1', composed of the N-terminal 259 amino acids.


Ritucharya: Answer to the lifestyle disorders.
Ritu, the season, classified by different features expresses different effects on the body as well as the environment. Ayurveda has depicted various rules and regimens (Charya), regarding diet and behavior to acclimatize seasonal enforcement easily without altering body homeostasis. The prime principle of Ayurvedic system of medicine is preventive aspect, can be achieved by the change in diet and practices in response to change in climatic condition. This is a very important aspect of preventive medicine as mentioned in Ayurvedic texts. Lifestyle disorders are very common in the present era, basically originating from lack of following seasonal regimens due to lack of concentration in seasonal characteristics. A firm scientific analysis is the base, which holds true even on date. In this review article, various regimens in diet and lifestyle as mentioned in the classics of Ayurveda and their importance on lifestyle disorders has been discussed.


[Induction of interferon-specific enzymes in cultures of cells sensitive and resistant to interferon].
Induction of 2'-5'-oligoadenylatesynthetase (2-5A synthetase) by interferons and theophylline by means of activation of cAMP-system in interferon susceptible and resistant cell lines were studied. In interferon resistant cell lines the basal activity of 2-5A synthetase exceeded the level of the same enzyme in interferon susceptible cell lines. Activity of 2-5A synthetase is increased in interferon susceptible cell lines by interferon treatment, but the activity of the enzyme is not altered in interferon resistant cell lines. Among the studied cell lines the induction of 2-5A synthetase by theophylline was possible only in L929 cell line. The common mechanism for the absence of 2-5A synthetase induction by interferon and theophylline in interferon resistant cells is discussed.


Advances in Doppler OCT.
We review the principle and some recent applications of Doppler optical coherence tomography (OCT). The advances of the phase-resolved Doppler OCT method are described. Functional OCT algorithms which are based on an extension of the phase-resolved scheme are also introduced. Recent applications of Doppler OCT for quantification of flow, imaging of microvasculature and vocal fold vibration, and optical coherence elastography are briefly discussed.


Molecular characteristics of carbapenem-resistant Raoultella ornithinolytica.
Aim of this study was to explore molecular characteristics and resistance mechanisms of carbapenem-resistant Raoultella ornithinolytica (CR-ROR) isolated from patients in a hospital in China. Three CR-ROR strains were collected and bacterial identification was done by Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) Vitek-MS and by digital DDH analysis. VITEK 2 compact system and Kirby-Bauer (K-B) disk diffusion were used for antimicrobial susceptibility testing. Whole genome sequencing was carried out using the Illumina platform NovaSeq sequencer. Abricate software was used for the prediction of antibiotic resistance genes of three CR-ROR strains. The phylogenetic tree was constructed through genome SNPs to investigate the genetic relationship of three CR-ROR strains. Three CR-ROR (WF1357, WF2441, and WF3367) strains were collected in this study. Two strains were isolated from neurosurgery (WF1357 and WF2441), and one was isolated from pulmonology department (WF3367). All strains harboured multiple antibiotic resistance genes. Two strains (WF1357, WF2441) carried the blaNDM-1 gene, one of the strains (WF3367) carried the blaKPC-2 gene. Three CR-RORs were resistant to different antimicrobial agents including carbapenems. The three CR-ROR strains collected in this study and 51 CR-ROR strain genomes downloaded from NCBI, were divided into six evolutionary groups (A-F). In this study, three CR-ROR strains were found to have a higher level of resistance to antibacterial agents and carried multiple antibiotic resistance genes. The CR-ROR strains carrying multiple antibacterial resistant genes require the stringent monitoring to avoid the spread of multidrug-resistant bacterial strains.


Randomized comparison of two anti-emetic strategies in high-risk patients undergoing day-case gynaecological surgery.
Postoperative nausea and vomiting (PONV) is a significant cause of morbidity among patients undergoing general anaesthesia. The optimal strategy for prevention of PONV, however, remains unclear. This study compared two commonly used prophylactic strategies in high-risk, day-case, gynaecological surgery patients. We conducted a randomized trial comparing sevoflurane combined with dolasetron (SD), with propofol-based total intravenous anaesthesia (TIVA) in 126 high-risk patients undergoing day-case gynaecological surgery. The primary endpoints included the incidence and severity of nausea or vomiting before discharge and the incidence of nausea or vomiting between discharge and 24 h. To identify the factors most predictive of a complete response (no PONV at any time within the 24 h period), multiple logistic regression models were fitted. Before discharge, there was no significant difference between the two treatment groups with respect to nausea and vomiting outcomes (P = 0.3). Post-discharge nausea and vomiting (PDNV), however, were significantly more common for patients in the TIVA group (nausea, P = 0.004 and vomiting, P = 0.03). Type of anaesthetic, adjusted for weight and anaesthesia duration was significantly associated with complete response (odds ratio = 2.7, 95% confidence interval = 1.15 to 6.4). Although both TIVA and dolasetron prophylaxis reduce the predicted rate of PONV in the early postoperative period, the anti-emetic effects of propofol are short-lived. A longer-acting drug such as dolasetron may therefore be necessary to prevent PDNV.


The NLRP3 inflammasome inhibitor OLT1177 rescues cognitive impairment in a mouse model of Alzheimer's disease.
Numerous studies demonstrate that neuroinflammation is a key player in the progression of Alzheimer's disease (AD). Interleukin (IL)-1β is a main inducer of inflammation and therefore a prime target for therapeutic options. The inactive IL-1β precursor requires processing by the the nucleotide-binding oligomerization domain-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome into a mature and active form. Studies have shown that IL-1β is up-regulated in brains of patients with AD, and that genetic inactivation of the NLRP3 inflammasome improves behavioral tests and synaptic plasticity phenotypes in a murine model of the disease. In the present study, we analyzed the effect of pharmacological inhibition of the NLRP3 inflammasome using dapansutrile (OLT1177), an oral NLRP3-specific inhibitor that is safe in humans. Six-month-old WT and APP/PS1 mice were fed with standard mouse chow or OLT1177-enriched chow for 3 mo. The Morris water maze test revealed an impaired learning and memory ability of 9-mo-old APP/PS1 mice (P = 0.001), which was completely rescued by OLT1177 fed to mice (P = 0.008 to untreated APP/PS1). Furthermore, our findings revealed that 3 mo of OLT1177 diet can rescue synaptic plasticity in this mouse model of AD (P = 0.007 to untreated APP/PS1). In addition, microglia were less activated (P = 0.07) and the number of plaques was reduced in the cortex (P = 0.03) following NLRP3 inhibition with OLT1177 administration. We also observed an OLT1177 dose-dependent normalization of plasma metabolic markers of AD to those of WT mice. This study suggests the therapeutic potential of treating neuroinflammation with an oral inhibitor of the NLRP3 inflammasome.


Iron replacement therapy in cancer-related anemia.
The incidence, etiology, and management of cancer-related anemia is reviewed and the role of i.v. iron therapy in its treatment is described. Between 30% and 90% of patients with cancer develop anemia due to direct effects of the disease, its treatment, underlying nutritional deficiencies, and the inflammation that characterizes chronic disease. Although the use of erythropoiesis-stimulating agents (ESAs) increases hemoglobin levels and decreases the need for transfusions, up to 50% of patients do not to respond to these drugs, usually due to the presence of absolute or functional iron deficiency. Multiple clinical trials have demonstrated that i.v. iron supplementation in patients with cancer-related anemia improves the response rate to ESAs, reduces the time to target hemoglobin levels, decreases ESA requirements, reduces costs, and is more efficacious than oral iron. These benefits are seen without increased toxicity. Nonetheless, i.v. iron remains underused in patients with cancer, partly due to misinformation and misinterpretation of the clinical nature of adverse events. Intravenous iron is underutilized in patients with cancer-related anemia. Based on published evidence, i.v. iron supplementation in patients with absolute or functional iron deficiency can improve patient responses to ESAs and reduce ESA requirements and may also reduce the need for transfusions and improve quality of life.


Effective connectivity inferred from fMRI transition dynamics during movie viewing points to a balanced reconfiguration of cortical interactions.
Our behavior entails a flexible and context-sensitive interplay between brain areas to integrate information according to goal-directed requirements. However, the neural mechanisms governing the entrainment of functionally specialized brain areas remain poorly understood. In particular, the question arises whether observed changes in the regional activity for different cognitive conditions are explained by modifications of the inputs to the brain or its connectivity? We observe that transitions of fMRI activity between areas convey information about the tasks performed by 19 subjects, watching a movie versus a black screen (rest). We use a model-based framework that explains this spatiotemporal functional connectivity pattern by the local variability for 66 cortical regions and the network effective connectivity between them. We find that, among the estimated model parameters, movie viewing affects to a larger extent the local activity, which we interpret as extrinsic changes related to the increased stimulus load. However, detailed changes in the effective connectivity preserve a balance in the propagating activity and select specific pathways such that high-level brain regions integrate visual and auditory information, in particular boosting the communication between the two brain hemispheres. These findings speak to a dynamic coordination underlying the functional integration in the brain.


Alpha-amylase supplementation of broiler diets based on corn.
A 42-d trial was conducted to study the influence of exogenous alpha-amylase on digestive and performance traits in broilers fed a corn-soybean meal diet. There were two treatments (control and alpha-amylase supplemented diet) and six replicates (14 Cobb male chicks caged together) per treatment. At 7 d of age, alpha-amylase supplementation improved daily gain by 9.4% (P < or = 0.05) and feed conversion by 4.2% (P < or = 0.01). At the end of the trial, birds fed the alpha-amylase-supplemented diet ate more and grew faster (P < or = 0.05) and hadbetter feed conversion (P < or = 0.10) than broilers fed the control diet. Also, alpha-amylase supplementation improved apparent fecal digestibility of organic matter and starch (P < or = 0.01) and AMEn of the diet (P < or = 0.001). However, no effects were detected for CP or fat digestibility. Nutrient digestibility and AMEn of the diet increased with age (P < or = 0.001); however, no interactions of alpha-amylase x age were observed for any trait. Coefficients of apparent ileal and fecal digestibility of starch at 28 d of age were similar, which indicated that most of the undigested starch was not fermented in the hindgut of the chick. alpha-Amylase supplementation reduced relative pancreas weight (P < or = 0.001) but did not affect the weight of the remaining organs. Age consistently reduced intestinal viscosity and relative weights of all the organs (P < or = 0.001). The data indicated that alpha-amylase supplementation of a corn-soybean meal diet improved digestibility of nutrients and performance of broilers.


Visual vs. Spectrophotometric Methods for Shade Selection.
The differences in tooth colour are important factors in the esthetic dentistry. The aim of this study was to determine the tooth colour using visual methods under natural light and "Easy Shade" device. Five hundred patients of Dental Clinic Graz Austria were selected for this study. The results of this study showed that the shade matching using "Easy Shade" device were not better than shade matching with visual methods under natural light. No difference was found between visual and digital methods in the selection of a tooth shade.


Cell-free synthesis and co-translational processing of the human asialoglycoprotein receptor.
The human asialoglycoprotein receptor is a 46-kDa membrane glycoprotein. It is initially synthesized as a 40-kDa precursor species possessing two N-linked high-mannose oligosaccharides which is subsequently converted to the 46-kDa mature product upon modification of its oligosaccharides of the complex form [Schwartz, A. L. & Rup, D. (1983) J. Biol. Chem. 258, 11 249-11 255]. To investigate further the biosynthesis of the human asialoglycoprotein receptor, we have utilized a cell-free wheat germ translation system supplemented with dog pancreatic microsomal membranes and programmed with HepG2 and human liver RNA. The primary translation product of the human receptor is a single 34-kDa species and this species is expressed throughout human fetal and adult development. The primary translation product possesses no cleavable signal peptide and is cotranslationally glycosylated to form the 40-kDa precursor species. In addition, the human asialoglycoprotein receptor is co-translationally inserted into microsomal membranes such that a 4-kDa cytoplasmic tail is susceptible to trypsin digestion.


Delineating the early dissemination mechanisms of acral melanoma by integrating single-cell and spatial transcriptomic analyses.
Acral melanoma (AM) is a rare subtype of melanoma characterized by a high incidence of lymph node (LN) metastasis, a critical factor in tumor dissemination and therapeutic decision-making. Here, we employ single-cell and spatial transcriptomic analyses to investigate the dynamic evolution of early AM dissemination. Our findings reveal substantial inter- and intra-tumor heterogeneity in AM, alongside a highly immunosuppressive tumor microenvironment and complex intercellular communication networks, particularly in patients with LN metastasis. Notably, we identify a strong association between MYC+ Melanoma (MYC+MEL) and FGFBP2+NKT cells with LN metastasis. Furthermore, we demonstrate that LN metastasis requires a metabolic shift towards fatty acid oxidation (FAO) induced by MITF in MYC+MEL cells. Etomoxir, a clinically approved FAO inhibitor, can effectively suppress MITF-mediated LN metastasis. This comprehensive dataset enhances our understanding of LN metastasis in AM, and provides insights into the potential therapeutic targeting for the management of early AM dissemination.


Kinetic properties of microtubule-activated 13 S and 21 S dynein ATPases. Evidence for allosteric behaviour associated with the inner row and outer row dynein arms.
The 13 S and 21 S dynein ATPases from Tetrahymena cilia rebind to extracted doublet microtubules as inner row and outer row arms. Rebinding is accompanied by four- to ninefold activation of the ATPase activity. The soluble (microtubule-free) forms of the two dyneins exhibit simple saturation kinetics (h = 1.0) with Vmax much less than mumol Pi mg-1 min-1 and Km = 20-40 microM-ATP. Mixing a fixed quantity of free dynein with increasing concentrations of extracted doublets results in systematic increases in all three kinetic parameters for each dynein. At infinite concentrations of doublets and ATP, each enzyme undergoes a significant shift to sigmoid saturation kinetics (h = 2-3), Vmax increases to a turnover rate of about 90 mol ATP per mol Es-1 and the Michaelis constant increases to much greater than 100 microM-ATP. These data suggest that both enzymes are allosteric and can be interpreted in terms of positive cooperativity relative to a minimum of two or three interacting sites. It is less clear whether this cooperativity is related to subunit interactions within the 21 S or 13 S particles, or to subunit interactions between adjacent particles (arms) on the microtubule lattice.


Influences of cardiorespiratory fitness levels and other predictors on cardiovascular disease mortality in men.
This investigation quantifies the relation between cardiorespiratory fitness levels and cardiovascular disease (CVD) mortality within strata of other CVD predictors. Participants included 25,341 male Cooper Clinic patients who underwent a maximal graded exercise test. CVD death rates were determined for low (least fit one-fifth), moderate (next two-fifths), and high (top two-fifths) cardiorespiratory fitness categories by strata of smoking habit, blood cholesterol level, resting blood pressure, and health status. There were 226 cardiovascular deaths during 211,996 man-years of follow-up. For individuals with none of the major CVD predictors (smoking, elevated resting systolic blood pressure, elevated blood cholesterol), there was a strong inverse relation (P = 0.001) between fitness level and CVD mortality. An inverse relation between CVD mortality and fitness level was seen within strata of cholesterol levels and health status. No evidence of a trend (P = 0.60) for decreased mortality was seen across fitness levels for individuals with elevated systolic blood pressure; however, a strong inverse gradient (P < 0.001) was seen across fitness levels for individuals with normal systolic blood pressure. There was a tendency for association between high levels of fitness and decreased CVD mortality in smokers compared with low and moderately fit smokers (P < 0.076). There was no significant association between level of fitness and CVD mortality for individuals with multiple (two or more) predictors (P = 0.325). Approximately 20% of the 226 CVD deaths in the population studied were attributed to low fitness level. Moderate and high levels of cardiorespiratory fitness seem to provide some protection from CVD mortality, even in the presence of well established CVD predictors.