Pain measurement in elders with chronic low back pain: traditional and alternative approaches.
Pain evaluation typically relies upon the use of self-report instruments. The validity of these tools is questionable in many older adults, however, particularly those with cognitive impairment. Rating of pain behavior (e.g. grimacing, sighing) by an objective observer represents an alternative pain assessment strategy which has been validated in subjects of heterogeneous ages. The purpose of this study was to examine, in a group of community-dwelling elderly with low back pain and lumbosacral osteoarthritis, the concurrent validity of observational pain behavior rating techniques as compared with self-report instruments and the degree to which pain and pain behavior relate to disability. Thirty-nine cognitively intact subjects, age > 65 years, without depression, other sources of pain, or other known spinal pathology underwent the following measures: (1) pain self-report using the verbal 0-10 scale, vertical verbal descriptor scale, Arthritis Impact Measurement Scales and McGill Pain Questionnaire; (2) pain behavior was sampled during two protocols, one, identical to that used by Keefe and Block (Behav. Ther., 13 (1982) 363-375), that required subjects to sit, stand, walk, and recline for 1-2 minute periods (which we have labelled the traditional protocol), and a second, more demanding protocol that was designed to simulate activities of daily living that place a premium on axial movement (the 'ADL' protocol); (3) disability was assessed using the Roland questionnaire, a 6 month global disability question and the Jette Functional Status Index; and (4) radiographic evaluation of the lumbosacral spine; osteoarthritis was quantitated using a previously validated scoring system. Interrelationships among pain, pain behavior and disability measures were tested using canonical correlations. Self-reported pain was associated with pain behavior frequency; the association was stronger when the ADL protocol was used, as compared with the traditional protocol. The association between pain and disability was modestly strong with both self-report instruments and pain behavior observation when the ADL protocol was used, but not when the traditional protocol was used. Our findings suggest that pain behavior observation is a valid assessment tool in the elderly. In addition, it seems that observation of elders during performance of activities of daily living may be a more sensitive and valid way of assessing pain behavior than observing pain behavior during sitting, walking, standing, or reclining.


Genetic deletion of α7 nicotinic acetylcholine receptors induces an age-dependent Alzheimer's disease-like pathology.
The accumulation of amyloid-beta peptide (Aβ) and the failure of cholinergic transmission are key players in Alzheimer's disease (AD). However, in the healthy brain, Aβ contributes to synaptic plasticity and memory acting through α7 subtype nicotinic acetylcholine receptors (α7nAChRs). Here, we hypothesized that the α7nAChR deletion blocks Aβ physiological function and promotes a compensatory increase in Aβ levels that, in turn, triggers an AD-like pathology. To validate this hypothesis, we studied the age-dependent phenotype of α7 knock out mice. We found that α7nAChR deletion caused an impairment of hippocampal synaptic plasticity and memory at 12 months of age, paralleled by an increase of Amyloid Precursor Protein expression and Aβ levels. This was accompanied by other classical AD features such as a hyperphosphorylation of tau at residues Ser 199, Ser 396, Thr 205, a decrease of GSK-3β at Ser 9, the presence of paired helical filaments and neurofibrillary tangles, neuronal loss and an increase of GFAP-positive astrocytes. Our findings suggest that α7nAChR malfunction might precede Aβ and tau pathology, offering a different perspective to interpret the failure of anti-Aβ therapies against AD and to find novel therapeutical approaches aimed at restoring α7nAChRs-mediated Aβ function at the synapse.


CD73: a novel target for cancer immunotherapy.
The promise of cancer immunotherapy has not been translated into clinical successes, in large part because of tumor-associated immune suppression that blocks effective antitumor immunity. Recent findings show a tumor-induced immunosuppressive mechanism, whereby tumor-derived CD73 functions as an ecto-enzyme to produce extracellular adenosine, which promotes tumor growth by limiting antitumor T-cell immunity via adenosine receptor signaling. Results with small molecule inhibitors, or monoclonal antibodies targeting CD73 in murine tumor models, suggest that targeted CD73 therapy is an important alternative and realistic approach to effective control of tumor growth. In particular, it helps T-cell-based therapy by enhancing the adaptive immune response machinery, which may increase the function of tumor-infiltrating T lymphocytes, and subsequently lead to improved survival in cancer patients.


Synthesis of a benzoxazinthione derivative of tanaproget and pharmacological evaluation for PET imaging of PR expression.
The histological evaluation of estrogen receptor (ER) and progesterone receptor (PR) expression in breast cancer lesions from biopsy tissue can stratify patients to receive endocrine therapy. Furthermore, PR expression can predict response to selective estrogen receptor modulators (SERMs). Current immunohistochemical approaches to PR detection are limited by sampling error associated with biopsy and lack of standardised protocols; positron emission tomography (PET) using receptor targeted radiopharmaceuticals to provide quantitative, whole-body imaging may overcome these limitations. PR expression has been successfully imaged with PET in the clinical setting, however investigation into new radioligands with improved pharmacokinetics and metabolic stability is desirable. We report the synthesis of a focused library of non-steroidal PR ligands evaluated for use as PET radioligands. A lead candidate ([18F]2) with low nanomolar activity was selected and radiolabelled with a radiochemical yield of 2.29 ± 2.31% (decay-corrected), radiochemical purity (RCP) > 95% and a molar activity of 2.5 ± 1.6 GBq/μmol. Cell uptake studies showed a significant and specific accumulation of [18F]2 in T47D (PR++) breast cancer cell compared to MDA-MB-231 (PR-) control; however, in vivo evaluation was confounded by rapid defluorination of the radioligand. In vitro metabolite analysis of 2 in MLM confirmed defluorination and oxidative metabolism of the thiocarbamate to oxocarbamate moiety by mass spectrometry. A route to access [18F]2 was developed to allow in vitro and in vivo evaluation, albeit with low radiochemical yield and modest molar activity. [18F]2 demonstrated selective uptake in PR++ T47D cells which could be blocked in a dose dependent manner with progesterone. However, [18F]2 showed poor in vivo metabolic stability with rapid defluorination within the time frame of the imaging protocol.


Air pollution exposure potentiates hypertension through reactive oxygen species-mediated activation of Rho/ROCK.
Fine particulate matter <2.5 microm (PM(2.5)) has been implicated in vasoconstriction and potentiation of hypertension in humans. We investigated the effects of short-term exposure to PM(2.5) in the angiotensin II (AII) infusion model. Sprague-Dawley rats were exposed to PM(2.5) or filtered air (FA) for 10 weeks. At week 9, minipumps containing AII were implanted and the responses studied over a week. Mean concentration of PM(2.5) inside the chamber was 79.1+/-7.4 microg/m(3). After AII infusion, mean arterial pressure was significantly higher in PM(2.5)-AII versus FA-AII group. Aortic vasoconstriction to phenylephrine was potentiated with exaggerated relaxation to the Rho-kinase (ROCK) inhibitor Y-27632 and increase in ROCK-1 mRNA levels in the PM(2.5)-AII group. Superoxide (O(2).(-)) production in aorta was increased in the PM(2.5)-AII compared to the FA group, inhibitable by apocynin and L-NAME with coordinate upregulation of NAD(P)H oxidase subunits p22(phox) and p47(phox) and depletion of tetrahydrobiopterin. In vitro exposure to ultrafine particles (UFP) and PM(2.5) was associated with an increase in ROCK activity, phosphorylation of myosin light chain, and myosin phosphatase target subunit (MYPT1). Pretreatment with the nonspecific antioxidant N-acetylcysteine and the Rho kinase inhibitors (Fasudil and Y-27632) prevented MLC and MYPT-1 phosphorylation by UFP suggesting a O(2)(.-)-mediated mechanism for PM(2.5) and UFP effects. Short-term air pollution exaggerates hypertension through O(2)(.-)-mediated upregulation of the Rho/ROCK pathway.


Case report. Adult Class II malocclusion with constricted arches, excessive vertical overlap and malposed teeth.
The patient presented with constriction of both arches, moderate crowding, posterior crossbite, severe deep bite, TMJ symptoms and facial pain. The Crozat appliance therapy effectively uprighted the posterior segments and developed the arches. Alignment, leveling and development of a good functional occlusion was accomplished efficiently with the straight-wire appliance. Facial balance and good lip support was maintained. No stripping was present and the periodontal health remains excellent. An improvement was made in the patient's smile and TMJ health. The teeth have been stable during the three year period following the initial placement of retainers. And the patient is very pleased with the treatment results.


Phosphagen and intracellular pH changes during contraction of creatine-depleted rat muscle.
To evaluate the functional role of phosphocreatine (PCr) and creatine in muscle metabolism, these compounds were depleted by feeding rats the creatine analogue, beta-guanidinopropionate (beta-GPA, 2% of diet). Changes in phosphate metabolites and intracellular pH were monitored in gastrocnemius muscle in situ by phosphorus nuclear magnetic resonance (31P-NMR) at 162 MHz using the surface coil technique. After 3 mo of feeding, 25 mumol/g of phosphorylated beta-GPA (beta-GPAP) had accumulated, and PCr, creatine, and ATP levels were reduced to 6, 17, and 56%, respectively, compared with muscles of control animals. In resting muscle, there was no measurable exchange of phosphate between beta-GPAP and ATP by the NMR saturation transfer method. During muscle stimulation at 1 and 5 Hz, the maximum net rate of beta-GPAP hydrolysis was 10% that of PCr in control muscles, so that after 150 s inorganic phosphate had increased to less than 50% of the level attained in control muscles. At both rates, peak twitch force declined toward a steady state more rapidly in beta-GPA-loaded muscles, but after 100 s force was either not different (1 Hz) or significantly greater (5 Hz) in the beta-GPA-fed animals. Intracellular pH initially decreased more rapidly during stimulation and recovered more rapidly afterward in the beta-GPA-loaded muscles compared with controls. This difference could be explained by the difference in expected proton consumption due to net PCr hydrolysis. However, despite buffering by PCr hydrolysis, pH ultimately decreased more in control muscle (6.1 vs. 6.3 for 5 Hz), indicating greater acid accumulation compared with beta-GPA-loaded muscles. In the superficial, predominantly fast-twitch glycolytic section of muscles clamp-frozen after 5-Hz stimulation for 150 s, lactate accumulation was twofold greater in controls. The results indicate that PCr is not essential for steady-state energy production but that the phosphate from PCr hydrolysis may be important for maximum activation of glycogenolysis and/or glycolysis.


Evidence-Based Health Informatics as the Foundation for the COVID-19 Response: A Joint Call for Action.
As a major public health crisis, the novel coronavirus disease 2019 (COVID-19) pandemic demonstrates the urgent need for safe, effective, and evidence-based implementations of digital health. The urgency stems from the frequent tendency to focus attention on seemingly high promising digital health interventions despite being poorly validated in times of crisis. In this paper, we describe a joint call for action to use and leverage evidence-based health informatics as the foundation for the COVID-19 response and public health interventions. Tangible examples are provided for how the working groups and special interest groups of the International Medical Informatics Association (IMIA) are helping to build an evidence-based response to this crisis. Leaders of working and special interest groups of the IMIA, a total of 26 groups, were contacted via e-mail to provide a summary of the scientific-based efforts taken to combat COVID-19 pandemic and participate in the discussion toward the creation of this manuscript. A total of 13 groups participated in this manuscript. Various efforts were exerted by members of IMIA including (1) developing evidence-based guidelines for the design and deployment of digital health solutions during COVID-19; (2) surveying clinical informaticians internationally about key digital solutions deployed to combat COVID-19 and the challenges faced when implementing and using them; and (3) offering necessary resources for clinicians about the use of digital tools in clinical practice, education, and research during COVID-19. Rigor and evidence need to be taken into consideration when designing, implementing, and using digital tools to combat COVID-19 to avoid delays and unforeseen negative consequences. It is paramount to employ a multidisciplinary approach for the development and implementation of digital health tools that have been rapidly deployed in response to the pandemic bearing in mind human factors, ethics, data privacy, and the diversity of context at the local, national, and international levels. The training and capacity building of front-line workers is crucial and must be linked to a clear strategy for evaluation of ongoing experiences.


Age and gender differences in the prevalence and patterns of multimorbidity in the older population.
The coexistence of several chronic diseases in one same individual, known as multimorbidity, is an important challenge facing health care systems in developed countries. Recent studies have revealed the existence of multimorbidity patterns clustering systematically associated distinct clinical entities. We sought to describe age and gender differences in the prevalence and patterns of multimorbidity in men and women over 65 years. Observational retrospective multicentre study based on diagnostic information gathered from electronic medical records of 19 primary care centres in Aragon and Catalonia. Multimorbidity patterns were identified through exploratory factor analysis. We performed a descriptive analysis of previously obtained patterns (i.e. cardiometabolic (CM), mechanical (MEC) and psychogeriatric (PG)) and the diseases included in the patterns stratifying by sex and age group. 67.5% of the aged population suffered two or more chronic diseases. 32.2% of men and 45.3% of women were assigned to at least one specific pattern of multimorbidity, and 4.6% of men and 8% of women presented more than one pattern simultaneously. Among women over 65 years the most frequent pattern was the MEC pattern (33.3%), whereas among men it was the CM pattern (21.2%). While the prevalence of the CM and MEC patterns decreased with age, the PG pattern showed a higher prevalence in the older age groups. Significant gender differences were observed in the prevalence of multimorbidity patterns, women showing a higher prevalence of the MEC and PG patterns, as well as a higher degree of pattern overlapping, probably due to a higher life expectancy and/or worse health. Future studies on multimorbidity patterns should take into account these differences and, therefore, the study of multimorbidity and its impact should be stratified by age and sex.


Absolute Monocyte and Platelet Counts May Provide Additional Prognostic Information in Primary Gastric Diffuse Large B-cell Lymphoma Patients Treated with Rituximab and CHOP.
Primary gastric diffuse large B cell lymphoma (PG-DLBCL) is the most common histological subtype of primary gastric lymphoma. The standard of care of PG-DLBCL patients is the combination rituximab-based immunochemotherapy (R-CHOP). Re-cently, different host-related factors have been shown to have significant prognostic significance in non-Hodgkin lymphoma. However, data regarding their prognostic contribution to PG-DLBCL are limited. To assess the prognostic impact of a panel of simple, cost-effective laboratory variables which are easy to apply in routine labora-tory use for R-CHOP-treated PG-DLBCL patients in an attempt to identify those among them that are high-risk category. We retrospectively assessed the possible prognostic impact of different laboratory markers in 42 R-CHOP treated PG-DLBCL patients treated between 2004 and 2014 and followed at a single institution. The estimated 5-year overall (OS) and progression-free survival (PFS) of the whole group were 80.9% and 78%, respectively. The absolute monocyte and platelet counts in univariate analysis predicted PFS and OS when analyzed as continuous and dichotomized variables. On multivariate analysis performed with factors included in the stage-modified International Prognostic Index (m-IPI), the absolute monocyte and platelet counts remained independent predictors of PFS and OS. Therefore, the absolute monocyte and platelet counts were combined to generate a prognostic index that identified patients with an especially poor overall survival. This prognostic index was independent of the m-IPI and could provide additional prognostic information for better stratification of these patients.


Pharmacokinetic Analysis of Intravenous Push Cefepime in Burn Patients with Augmented Renal Clearance.
Patients with augmented renal clearance (ARC) are a subset of critically ill patients including burn patients that exhibit increased renal elimination of medications beyond that of similarly injured patients. Currently approved maximum regimens of medications primarily eliminated by the kidney, such as cefepime (>90% unchanged in the urine), may be inadequate (e.g. compromising the bactericidal activity of cefepime) in patients with ARC. Due to recent resource limitations, centers have changed infusion practices of commonly prescribed medications to intravenous push (IVP), potentially exacerbating the problem of maintaining bactericidal cefepime concentrations. The hypothesis of the study was patients with ARC are not currently achieving adequate target attainment, when receiving cefepime 2g every 8 hours IVP. Eight blood samples were collected from each patient, and concentrations measured via LC-MS/MS. WinNonlin (version 8.3) was used to estimate the pharmacokinetic parameters of cefepime and simulate plasma concentrations of cefepime in each of the ten subjects. Simulations of cefepime plasma concentrations produced by a 2 g dose given every 8 hours and a 1 g dose given every 4 hours were performed and the time above a MIC of 4 mg/L, 8 mg/L, and 16 mg/L compared. The 2 g every 8 hours regimen remained above the breakpoints for 92%, 85%, and 71% of the dosing interval, respectively. The 1 g every 4 hours regimen remained above the same breakpoints at a frequency of 100%, 99%, and 92% of the dosing interval. Giving cefepime 1 g every 4 hours is a simple approach to increase the likelihood of maintaining the optimal bactericidal activity of cefepime in patients with ARC.


Outpatient continuing care after residential substance use disorder treatment in the US Veterans Health Administration: Facilitators and challenges.
In the U.S. Veterans Health Administration (VHA), residential treatment programs are an important part of the continuum of care for patients with substance use disorders (SUDs). Outpatient continuing care after residential treatment helps maintain early recovery and treatment gains. Knowing more about the drivers of variation in continuing care practices and performance across residential programs might inform quality improvement efforts. Metrics of continuing care were operationalized and calculated for each of VHA's 63 SUD Residential Rehabilitation Treatment Programs (SUD RRTPs) and 34 Mental Health Residential Rehabilitation Treatment Programs (MH RRTPs) with a SUD track in fiscal year 2012. Management and frontline staff of these programs were then interviewed to learn what factors might contribute to high or low program performance on the metrics compared with national averages. Among SUD RRTPs, the mean rate of outpatient SUD/MH continuing care was 59% within 7 days and 80% within 30 days, and the mean rate of SUD continuing care was 63% within 30 days. Among MH RRTPs with a SUD track, these rates were 56%, 75%, and 36%, respectively. There was substantial variability in continuing care rates across the 97 programs: 21%-93% for SUD/MH care within 7 days, 36%-100% for SUD/MH care within 30 days, and 4%-91% for SUD care within 30 days. Interviews with representatives of 44 programs revealed key facilitators of continuing care: accountability of program staff, predischarge scheduling, predischarge introductions to continuing care providers, strong patient relationships, accessibility, and persistent emphasis. Key challenges included inadequate program staffing, lack of program staff accountability, and poor accessibility. Wide variation in continuing care rates across programs and identification of common facilitators at high-performing programs suggest substantial opportunity for improvement for programs with lower performance.


Empathy as a Concept from Bench to Bedside: A Translational Challenge.
Empathy is a multidimensional paradigm, and there currently is a lack of scientific consensus in its definition. In this paper, we review the possibility of compromising data during behavioral neuroscience experiments, including but not limited to those who study empathy. The experimental protocols can affect, and be affected by, empathy and related processes at multiple levels. We discuss several points to help researchers develop a successful translational pathway for behavioral research on empathy. Despite varying in their focus with no widely accepted model, current rodent models on empathy have provided sound translational explanations for many neuropsychiatric proof-of-concepts to date. Research has shown that empathy can be influenced by many parameters, some of which are to be reviewed in this paper. We emphasize the future importance of consistency in modeling proof of concept; efforts to create a multidisciplinary group which would include both bench scientists and clinicians with expertise in neuropsychiatry, and the consideration of empathy as an independent variable in animal behavioral experimental designs which is not the mainstream practice at present.


Relation between structure and immunologic properties of the Vi capsular polysaccharide.
The Vi capsular polysaccharide of Salmonella typhi is a linear homopolymer of poly-alpha(1----4)GalNAcp variably O acetylated at the C-3 position. Serum antibodies elicited by this antigen confer protective immunity against typhoid fever. The relation between the immunologic properties and structure of Vi was investigated by carboxyl reduction, O deacetylation, and acid hydrolysis. The immunogenicity of Vi was closely related to its degree of O acetylation. Partial O deacetylation slightly increased immunogenicity; complete O deacetylation eliminated the immunogenicity of Vi. O-deacetylated Vi, however, still reacted with antisera prepared by injection of whole bacteria. Carboxyl reduction, in contrast, had a comparatively slight effect upon both the immunogenicity and antigenicity of Vi. Retention levels of antigenicity after acid treatment were greater for both the native and carboxyl-reduced Vi than for the O-deacetylated product. The Courtauld-Koltun space-filling model of a pentamer of Vi demonstrated that the bulky nonpolar O-acetyls, which protrude in rows on both sides, make up most of the surface. The carboxyls are less exposed and are partially shielded by the O-acetyls. The molecular model thus provides an explantation for the dominant role of the O-acetyls, as well as the lesser effect of carboxyl reduction, upon the immunologic properties of Vi.


Tn554 inserts in methicillin-resistant Staphylococcus aureus from Australia and England: comparison with an American methicillin-resistant group.
We have compared methicillin-resistant (Mcr) Staphylococcus aureus isolates from Australia, the UK and the USA with regard to chromosomal inserts of the macrolides-lincosamides-streptogramin B (MLS)-resistance transposon Tn554. The American isolates were known to have a distinctive Tn554 insert, designated insert 6, which was closely associated epidemiologically with the methicillin-resistance phenotype. Southern blots of DNA from Australian and London, UK Mcr isolates were hybridized with a range of probes related to Tn554. The isolates had similar or identical Tn554 inserts, and we consider them to be a single group, designated 'Australondon'. Australondon isolates were compared in detail with a deletion mutant, ANS62, that had lost the methicillin-resistance determinant mec, plus other resistance determinants resident in the mec region of the chromosome, and with an American Mcr isolate containing Tn554 insert 6. The Australondon isolates had three Tn554 inserts. Sequence analysis with the polymerase chain reaction showed that all of these inserts differed from classical Tn554 in that the 3'-terminal residues of the transposons were reverse complements of the usual GATGTA. One of the Australondon inserts, designated 6B, closely resembled Tn554 insert 6 in the sequence of its left flanking chromosomal DNA. This insert was found to abut the deletion from the mec region which results in strain ANS62. We infer that Tn554 insert 6B is part of the mec region of the chromosome in Australondon isolates, supporting the idea that insert 6 of the American isolates is also part of this chromosomal region.


Incidentally Discovered Unruptured AICA Aneurysm After Radiosurgery for Vestibular Schwannoma: A Case Report and Review of the Literature.
This is a case report and review of the literature of aneurysm formation after stereotactic radiosurgery (SRS) in the posterior fossa. Cerebral aneurysm formation is not a commonly recognized complication of SRS. We present the first case of an unruptured anteroinferior cerebellar artery aneurysm incidentally found at surgery in a patient with trigeminal neuralgia secondary to a vestibular schwannoma (VS) first treated with Gamma Knife radiosurgery. Other cases of posterior fossa aneurysms associated with SRS and the pathogenesis of vascular injury by radiation are discussed. A 57-year-old woman with medically intractable severe trigeminal neuralgia secondary to a 1.4-cm VS treated with SRS 10 years previously at an outside institution. The patient underwent a left retrosigmoid craniotomy for tumor debulking. During resection, two small aneurysms on the tumor's ventral side arising from the main trunk of the anteroinferior cerebellar artery were encountered and treated with direct clip ligation, sparing the parent vessel. The patient did well after surgery and was discharged home on Hospital Day 4 at her neurologic baseline, with normal facial nerve function and without trigeminal pain. Although aneurysms associated with posterior fossa SRS are rare, there are at least seven reports, including the current case, in the past decade. Because the relationship between radiation and aneurysm formation is unproven and controversial, further study, especially examining long-term effects, is needed. Given the overall rarity and uncertain association between SRS and aneurysm formation, we do not recommend routine aneurysm surveillance screening in patients undergoing Gamma Knife radiosurgery for VS. Surgeons should be aware of the rare possibility of encountering an aneurysm during surgical exploration in patients with VS who fail SRS.


Economic value evaluation in disease management programs.
Chronic disease management has been a rapidly growing entity in the 21st century as a strategy for managing chronic illnesses in large populations. However, experience has shown that disease management programs have not been able to demonstrate their financial value. The objectives of disease management programs are to create quality benchmarks, such as principles and guidelines, and to establish a uniform set of metrics and a standardized methodology for evaluating them. In order to illuminate the essence of disease management and its components, as well as the complexity and the problematic nature of performing economic calculations of their profitability and value, we collected data from several reports that dealt with the economic intervention of disease management programs. The disease management economic evaluation is composed of a series of steps, including the following major categories: data/information technology, information generation, assessment/recommendations, actionable customer plans, and program assessment/reassessment. We demonstrate the elements necessary for economic analysis. Disease management is one of the most innovative tools in the managed care environment and is still in the process of being defined. Therefore, objectives should include the creation of quality measures, such as principles and guidelines, and the establishment of a uniform set of metrics and a standardized methodology for evaluating them.


Characterization of cis-elements required for osmotic response of rat Na(+)/H(+) exchanger-2 (NHE-2) gene.
The Na(+)/H(+) exchanger (NHE-2) has been implicated in osmoregulation in the kidney, because it transports Na(+) across the cell membrane and efficiently alters intracellular osmolarity. On hyperosmotic stress, NHE-2 mRNA increases in abundance in mouse inner medullary collecting duct (mIMCD-3) cells, suggesting possible transcriptional regulation. To investigate the molecular mechanism of potential transcriptional regulation of NHE-2 by hyperosmolarity, we have functionally characterized the 5'-flanking region of the gene in mIMCD-3 cells. Transient transfection of luciferase reporter gene constructs revealed a novel cis-acting element, which we call OsmoE (osmotic-responsive element, bp -808 to -791, GGGCCAGTTGGCGCTGGG), and a TonE-like element (tonicity-responsive element, bp -1201 to -1189, GCTGGAAAACCGA), which together are shown to be responsible for hyperosmotic induction of the NHE-2 gene. Electrophoretic mobility shift assays suggest that different DNA-protein interactions occur between these two osmotic response elements. However, both DNA sequences were shown to specifically bind nuclear proteins that dramatically increase in abundance under hyperosmotic conditions. Isolation of trans-acting factors and characterization of their specific interaction with these osmotic response elements will further elucidate the transcriptional mechanisms controlling NHE-2 gene expression under hyperosmolar conditions.


A polymorphism of the methionine synthase reductase gene increases chromosomal damage in peripheral lymphocytes in smokers.
The cytogenetic effects of cigarette smoke has been evaluated as one of many potential confounders in a large number of biomonitoring studies of occupationally or environmentally exposed populations and control subjects. Despite the well-known presence of carcinogens in the cigarette smoke, the results in the scientific literature linking smoking habits to micronuclei (MN) frequency, one of the cytogenetic markers, are rather controversial. Here, we investigated the relationships among MN frequency, smoking habits and five folate metabolic enzyme gene polymorphisms (MTHFR C677T and A1298C, MTR A2756G, MTRR A66G and TYMS 3'UTR) in 132 healthy Japanese men who were non-habitual drinkers. In never- and former-smokers, no statistically significant differences in the mean MN frequencies were observed according to the five folate metabolic enzyme gene polymorphisms. In current-smokers, however, subjects with the AA genotype for MTRR had a significantly higher mean MN frequency than the AG genotypes for MTRR (p<0.05). Furthermore, among subjects with the AA genotype for MTRR, current-smokers were found to have a significantly higher mean MN frequency than never- and former-smokers (p<0.05). To further characterize this association, we stratified the smoking status into five groups: non-smokers (never-smokers and former-smokers), 1-10 cigarettes/day, 11-20 cigarettes/day, 21-30 cigarettes/day and >or=31 cigarettes/day. There was an overall trend for the mean MN frequency in subjects with the MTRR AA genotype to increase as the number of cigarettes smoked per day increased (p<0.01, Jonckheere-Terpstra test). The results of our preliminary study suggest that the MTRR AA genotype acts to increase the MN frequency resulting from cigarette smoking. Therefore, studies on human genotoxicity based on cytogenetic markers of MN should take into account both the MTRR polymorphism and the potential confounding effect of smoking, although these preliminary findings need to be validated in larger populations because of the relatively small sample size.


[Examination of acute phase proteins concentrations in children with allergic rhinitis].
Allergic rhinitis is an inflammatory disorder of upper respiratory tract (about 15 per cent of the population in industrialized countries suffer from this condition), characterized by frequent sneezing and a runny or stuffy nose sometimes accompanied by watery eyes. As the most common allergic condition, allergic rhinitis affects people of all ages. Boys are twice as likely to get allergic rhinitis than girls. The median age of onset of the condition is 10 years old, meaning that equal numbers of children develop the condition before and after age 10. Symptoms usually appear in childhood first and then lessen by the age of 30 or 40. Seasonal allergic rhinitis usually results from tree, grass or weed pollen. With this type of rhinitis, symptoms will decrease with the arrival of cold weather. Perennial allergic rhinitis can cause year-round symptoms. This allergic reaction is the result of indoor irritants such as feathers, mold spores, animal dander (hair and skin shed by pets) or dust mites. It is often aggravated by a food allergy, the most common being an allergy to milk. Acute phase proteins (APP) belong to the most ancient part of the unspecific immunity and contribute markedly to the keeping of homeostasis. As much as 30 various proteins are for the moment regarded as APP. Being multifunctional regulators and effectors APP stay in multiple relations to practically all types of cells and molecules. Among APP following functional groups may be described: transport proteins (transferrin, ceruloplasmin and haptoglobin), clotting factors (fibrinogen), antiproteases (alpha1-antitrypsin, alpha1-antichymotrypsin, alpha2-macroglobulin), complement components (C3, C4) and several proteins of hardly known function, like C-reactive protein (CRP), serum amyloid A, acid alpha1-glycoprotein (AGP) and others. From a group of 32 children, aged from 5 to 14 years, with symptoms of seasonal allergic rhinitis, and from a control group of 10 healthy children sex and age matched, blood sample was taken after written parents' consent. In all children skin tests (Stallergens) were performed at the beginning of the year. Any reaction was present in the control group. Following proteins were analyzed: CRP, AGP, alpha1-antichymotrypsin (ACT), transferrin, ceruloplasmin (Cp), alpha1-antitrypsin, haptoglobin and alpha2-macroglobulin (A2M). C-reactive protein level was very low, what allowed us to exclude all acute conditions. As expected, AGP and ACT concentrations were not elevated, either, and even non significantly lower values were observed in allergic children in comparison to controls. It is worth noticing that transferrin concentration was statistically lower in allergic children, as well as A2M and Cp concentrations. All this suggest an on-going disturbance in cytokine network that may directly affect both the iron metabolism and the non-specific immunity. It may be stated that allergic rhinitis causes impairment of acute phase proteins synthesis, which may affect natural defense or homeostasis in the immune system of a child.


Ovine and Human Placental IgG inhibit Human Natural Killer Cell Cytotoxicity in vitro.
To determine the effect of ovine and human placental IgG on human Natural Killer (NK) cell cytotoxicity in vitro placental IgG was eluted at acidic pH and purified by ion exchange and subsequently by affinity chromatography on protein G and protein A sepharose columns. These antibodies were analysed for presense of IgG by immuno-electrophoresis and relative purity determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS PAGE). The effect of these antibodies on human NK cell cytotoxicity was investigated by slChromium Release Assay using human K562 cells as targets and human peripheral blood lymphocytes as effector cells. Both ovine and human placental IgG inhibited human NK cell cytotoxicity in a dose dependent manner. Placental IgG may down-regulate the cytotoxic effects of NK cells in vivo by competitively excluding the binding of NK cells to their respective targets on the trophoblast. Alternatively, these antibodies may themselves be toxic to NK cells. Either way, the presence of these antibodies on the placental trophoblast may prevent the binding of NK cells and subsequent immunological rejection of the fetal allograft. Also, ovine placental IgG may be functionally similar to its human counterpart and may therefore be suitable as a model for the study of maternal fetal interaction during pregnancy in humans.


Converting biomechanical energy into electricity by a muscle-movement-driven nanogenerator.
A living species has numerous sources of mechanical energy, such as muscle stretching, arm/leg swings, walking/running, heart beats, and blood flow. We demonstrate a piezoelectric nanowire based nanogenerator that converts biomechanical energy, such as the movement of a human finger and the body motion of a live hamster (Campbell's dwarf), into electricity. A single wire generator (SWG) consists of a flexible substrate with a ZnO nanowire affixed laterally at its two ends on the substrate surface. Muscle stretching results in the back and forth stretching of the substrate and the nanowire. The piezoelectric potential created inside the wire leads to the flow of electrons in the external circuit. The output voltage has been increased by integrating multiple SWGs. A series connection of four SWGs produced an output voltage of up to approximately 0.1-0.15 V. The success of energy harvesting from a tapping finger and a running hamster reveals the potential of using the nanogenerators for scavenging low-frequency energy from regular and irregular biomotion.


Creating Whole Person Health Care Systems: Understanding Employee Perceptions of VAs Whole Health Cultural Transformation.
Objective: Whole person health care, like that being implemented in the U.S. Veterans Health Administration (VHA), involves person-centered approaches that address what matters most to patients to achieve well-being beyond the biomedical absence of disease. As whole health (WH) approaches expand, their integration into clinical practice is predicated on health care employees reconceptualizing practice beyond find-it-fix-it medicine and embracing WH as a new philosophy of care. This study examined employee perspectives of WH and their integration of this approach into care. Design: We conducted a survey with responses from 1073 clinical and 800 nonclinical employees at 5 VHA WH Flagship sites about their perceptions and use of a WH approach. We used descriptive statistics to examine employees' support for WH and conducted thematic analysis to qualitatively explore their perceptions about this approach from free-text comments supplied by 475 respondents. Results: On structured survey items, employees largely agreed that WH was a valuable approach but were relatively less likely to have incorporated it into practice or report support within their organization for WH. Qualitative comments revealed varying conceptualizations of WH. While some respondents understood that WH represented a philosophical shift in care, many characterized WH narrowly as services. These conceptualizations contributed to lower perceived relevance, skepticism, and misgivings that WH diverted needed resources away from existing clinical services. Organizational context including leadership messaging, siloed structures, and limited educational opportunities reinforced these perceptions. Conclusions: Successfully transforming the culture of care requires a shift in mindset among employees and leadership alike. Employees' depictions didn't always reflect WH as a person-centered approach designed to engage patients to enhance their health and well-being. Without consistent leadership messaging and accessible training, opportunities to expand understandings of WH are likely to be missed. To promote WH transformation, additional attention is needed for employees to embrace this approach to care.


The effect of bone loss on rod-like and plate-like trabeculae in the cancellous bone of the mandibular condyle.
Bone loss may affect the structure of cancellous bone. But its effect on trabeculae with different characteristics, like rods and plates, is not accurately known. This study analyzes the effect of bone loss on individual rod-like and plate-like trabeculae. 94 specimens were obtained from mandibular condyles from both dentate and edentate humans and scanned with a micro-CT scanner. The bone volume fraction (BV/TV) of these specimens ranged from 7% to 30%. Next, the rod-like and plate-like trabeculae were identified with an especially developed algorithm. Plate volume fraction (PV/TV), rod volume fraction (RV/TV), plate thickness, rod thickness, number of plates, and number of rods were determined. In individual specimens, the thickness of the rods ranged from 40 mum to 180 mum, while the thickness of the plates ranged from 40 mum to 300 mum. In every specimen, the thickness of the plates was larger than the thickness of the rods. Statistical analysis revealed that PV/TV was proportional (r = 0.98, P < 0.001) and RV/TV inversely proportional (r = -0.45, P < 0.001) to BV/TV. Also the thickness of the plates correlated with BV/TV (r = 0.62, P < 0.001), while the thickness of the rods (mean = 90 mum, SD = 7 mum) remained constant (r = -0.09, P = 0.378). A four-fold reduction of the bone volume fraction was accompanied by a three-fold reduction of the number of plates and a 40% reduction of their thickness, but also by a three-fold increase in the number of rods and RV/TV. It was concluded that the effect of bone loss on plate-like trabeculae was opposite to its effect on rod-like trabeculae. Remarkably, the thickness of the rods (90 mum) was independent of the bone volume fraction. This suggests that there is a minimal thickness for trabeculae.


Considerations and Outcomes for Adolescents and Young Adults With Cloacal Anomalies: A Scoping Review of Urologic, Colorectal, Gynecologic and Psychosocial Concerns.
The objective of this scoping review is to provide a summary of the current literature regarding adolescents and young adults with histories of cloacal anomalies. Preferred Reporting Items for Systematic Reviews and Meta-analysis Extension for Scoping Reviews were used. Data were categorized into four domains-urologic, colorectal, gynecologic/obstetric, and sexual/psychosocial. The current literature has poor study quality and mostly consists of retrospective studies of small cohorts with varying definitions of outcomes. Women with cloacal anomalies are at high risk for urologic dysfunction but can maintain kidney health and achieve social continence with medical and surgical management. Sexual function and adult healthcare transition are areas ripe for improved future research.


Inhaled interferons beta and SARS-COV2 infection: a preliminary therapeutic perspective.
SARS-COV2 infection represents a therapeutic challenge due to the limited number of effective therapies available and due to the fact that it is not clear which host response in terms of inflammation pattern is the most predictive for an optimal (and rapid) recovery. Interferon β pathway is impaired in SARS-COV2 infection and this is associated with a bigger disease burden. Exogenous inhaled interferon might be beneficial in this setting. Nebulized interferon-β is currently investigated as a potential therapy for SARS-COV2 because the available data from a phase II study demonstrate that this medication is able to accelerate the recovery from disease. Further clinical studies are needed in order to better document the efficacy of this therapy especially in severe forms of COVID-19, the optimal duration of therapy and if such a medication is appropriate for domiciliary use. Also combined regimens with antivirals or with compounds which are able to enhance the endogenous production of interferon might be of promise.


Open adrenalectomy.
In an era when minimally invasive adrenalectomy is the gold standard treatment for majority of patients presenting with adrenal tumours, open adrenalectomy has become an operation that should be centralised in regional referral centers. Its main indication is represented by patients with large (>8 cm) phaeochromocytomas and patients with cortical adrenal tumours suspected of malignancy either because of their size (>4-6 cm) or because of radiological appearance of local invasion. Based on local expertise some of these patients might benefit from multidisciplinary input from liver or transplant surgeons. This chapter will discuss the anatomical landmarks and will comment on different steps in the procedure for right- or left-sided procedure. It is outside the scope of this chapter to settle the ongoing debate about patient selection for laparoscopic or open adrenalectomy when the diagnosis of adrenocortical cancer is suspected preoperatively.


Lack of efficacy of tacrolimus ointment 0.1% for treatment of hemodialysis-related pruritus: a randomized, double-blind, vehicle-controlled study.
Pruritus is a common and disabling symptom for patients undergoing hemodialysis. Many topical and systemic treatments have been used for uremic pruritus, mostly in anecdotal reports. A recent case series demonstrated that topical tacrolimus ointment 0.03% had a significant antipruritic effect for patients with uremia. In an attempt to confirm these findings, we conducted a randomized, double-blind, vehicle-controlled study to assess the efficacy of tacrolimus ointment 0.1% for the treatment of hemodialysis-related pruritus. The results of this study do not demonstrate that tacrolimus ointment 0.1% is more effective than vehicle in relieving uremic pruritus.


Glucose metabolism-targeted therapy and withaferin A are effective for epidermal growth factor receptor tyrosine kinase inhibitor-induced drug-tolerant persisters.
In pathway-targeted cancer drug therapies, the relatively rapid emergence of drug-tolerant persisters (DTPs) substantially limits the overall therapeutic benefit. However, little is known about the roles of DTPs in drug resistance. In this study, we investigated the features of epidermal growth factor receptor-tyrosine kinase inhibitor-induced DTPs and explored a new treatment strategy to overcome the emergence of these DTPs. We used two EGFR-mutated lung adenocarcinoma cell lines, PC9 and II-18. They were treated with 2 μM gefitinib for 6, 12, or 24 days or 6 months. We analyzed the mRNA expression of the stem cell-related markers by quantitative RT-PCR and the expression of the cellular senescence-associated proteins. Then we sorted DTPs according to the expression pattern of CD133 and analyzed the features of sorted cells. Finally, we tried to ablate DTPs by glucose metabolism targeting therapies and a stem-like cell targeting drug, withaferin A. Drug-tolerant persisters were composed of at least two types of cells, one with the properties of cancer stem-like cells (CSCs) and the other with the properties of therapy-induced senescent (TIS) cells. The CD133high cell population had CSC properties and the CD133low cell population had TIS properties. The CD133low cell population containing TIS cells showed a senescence-associated secretory phenotype that supported the emergence of the CD133high cell population containing CSCs. Glucose metabolism inhibitors effectively eliminated the CD133low cell population. Withaferin A effectively eliminated the CD133high cell population. The combination of phloretin and withaferin A effectively suppressed gefitinib-resistant tumor growth.


Asiatic acid inhibits LPS-induced inflammatory response in human gingival fibroblasts.
Asiatic acid, a triterpenoid component isolated from Centella asiatica (L.) Urban, possesses antioxidative and anti-inflammatory activities. In this study, we aimed to investigate the anti-inflammatory effects of asiatic acid both in vivo and in vitro. HGFs or RAW264.7 cells were treated with asiatic acid 1h before LPS treatment. Cell viability was measured by MTT assay. The levels of PGE2, NO, IL-6, and IL-8 were detected by ELISA. Protein expression levels were detected by western blot analysis. In vivo, asiatic acid significantly inhibited LPS-induced IL-6 and IL-8 expression levels in gingival tissues. In vitro, LPS-induced PGE2, NO, IL-6, and IL-8 production was significantly attenuated by asiatic acid. Asiatic acid also inhibited p65 NF-κB phosphorylation induced by LPS in HGFs. The expression of PPAR-γ was up-regulated by asiatic acid. Furthermore, GW9662, a PPAR-γ inhibitor, attenuated the inhibitory effect of asiatic acid on PGE2, NO, IL-6, and IL-8 production. Our results suggest that asiatic acid activates PPAR-γ, which subsequently inhibits LPS-induced NF-κB activation and inflammatory mediators production. Asiatic acid may offer therapeutic potential for the treatment of periodontitis.


Overlapping vs. one long stenting in long coronary lesions.
Intervention of long coronary lesions remains problematic, and optimal treatment strategy is yet to be determined. Despite advancement of stent technology, data are few regarding the efficacy of overlapping stents vs. a single long stent in long coronary lesions. This study was performed to evaluate the results of those strategies for long coronary lesions and to determine the predictors of in-stent restenosis (ISR). Sixty-four lesions (> 20 mm) in 64 patients were treated with either one long stent (group 1, n = 32) or two overlapping stents (group 2, n = 32). Overlapping stents were used at tortuous or calcified lesions and at lesions with diameter discrepancy or significant dissection. Immediate results, follow-up clinical and angiographic outcomes, and predictors of ISR were evaluated. Procedures were successful in all patients in both groups. Clinical and angiographic follow-ups were performed in 54 (84%) cases and 50 (78%) cases, respectively. During the follow-up, major adverse cardiac event occurred in 36% of group 1 and 29% of group 2 (P = 0.56). Six-month ISR rates were 39% in group 1 and 41% in group 2 (P = 0.91). Age (>/= 65 years old) was an independent risk factor of ISR (54% vs. 23%; OR = 4.4; P = 0.04), and distal reference diameter (RD) of less than 2.5 mm tended to predict ISR in multivariate analysis (60% vs. 25%; OR = 3.5; P = 0.06). In conclusion, stent overlapping can be used with outcome similar to that of one long stent in long coronary lesions. The optimal result may be obtained by considering the patient's age and the distal vessel diameter of the lesion.


Safety and toxicity of nonoxynol-9 gel as a rectal microbicide.
Methods of HIV and STD prevention, which can be controlled by the receptive partner, are a high priority for research and development. Studies on the safety of Nonoxynol-9 (N-9) on the vaginal mucosa have yielded conflicting results. No Phase I study has evaluated the effect of N-9 on the rectal mucosa. To assess the safety of 52.5 mg of N-9 in a 1.5-g gel when applied one to four applicators per day to the rectum and penis. The study included 25 HIV-negative and 10 HIV-positive, monogamous gay male couples in which each partner was exclusively insertive or receptive while using N-9 gel. Each participant served as his own control during placebo gel use compared to during N-9 gel use. Receptive partners underwent anoscopic examination after 1 week of placebo use and after 2, 5, and 6 weeks of N-9 gel use, with rectal biopsies obtained after 1 week of placebo use and after 5 and 6 weeks of N-9 gel use. Insertive partners had safety monitoring after 1 week of placebo use and after 2, 5, and 6 weeks of N-9. No rectal ulcers were detected; superficial rectal erosions were noted in two HIV-negative participants. Abnormal or slightly abnormal histologic abnormalities of rectal biopsies were detected in 31 (89%) receptive participants after N-9 gel use compared to 24 (69%) participants after 1 week of placebo gel use. Meatal ulceration, not caused by herpes simplex virus, was detected in one HIV-negative participant. Low-dose N-9 gel was not associated with macroscopic rectal and penile epithelial disruption or inflammation, but histologic abnormalities were commonly observed during N-9 gel as well as during placebo gel use.


Evaluation of dexmedetomidine and ketamine in combination with opioids as injectable anesthesia for castration in dogs.
To compare efficacy and cardiorespiratory effects of dexmedetomidine and ketamine in combination with butorphanol, hydromorphone, or buprenorphine (with or without reversal by atipamezole) in dogs undergoing castration. Prospective, randomized, split-plot, blinded study. 30 healthy client-owned sexually intact male dogs. Dogs (n = 10 dogs/group) were assigned to receive dexmedetomidine (15 μg/kg [6.82 μg/lb]) and ketamine (3 mg/kg [1.36 mg/lb]) with butorphanol (0.2 mg/kg [0.09 mg/lb]; DKBut), the same dosages of dexmedetomidine and ketamine with hydromorphone (0.05 mg/kg [0.023 mg/lb]; DKH), or the same dosages of dexmedetomidine and ketamine with buprenorphine (40 μg/kg [18.18 μg/lb]; DKBup). All drugs were administered as a single IM injection for induction and maintenance of anesthesia for castration. At conclusion of the surgery, 5 dogs in each treatment group received atipamezole (150 μg/kg [68.18 μg/lb], IM), and the remainder received saline (0.9% NaCl) solution IM. Cardiorespiratory variables and quality of anesthesia were assessed. Supplemental isoflurane was administered to the dogs when anesthesia was considered inadequate during surgery. All drug combinations rapidly induced anesthesia. Dogs were intubated within 10 minutes after injection. Supplemental isoflurane was needed during surgery in 1, 3, and 4 dogs in the DKBup, DKBut, and DKH groups, respectively. Dogs that received atipamezole had a significantly shorter recovery time. Some dogs in each group had bradycardia and hypoxemia with hypertension. DKBup was the most suitable injectable anesthetic combination used. Recovery was shortened by IM administration of atipamezole. There were minimal adverse effects in all groups.


Outcomes of reintervention for recurrent symptomatic disease after tibial endovascular intervention.
Tibial interventions for critical limb ischemia are now commonplace. Restenosis and occlusion remain barriers to durability after intervention. The aim of this study was to examine the patient-centered outcomes of open and endovascular reintervention for symptomatic recurrent disease after a primary isolated tibial endovascular intervention. A database of patients undergoing isolated primary lower extremity tibial endovascular interventions between 2006 and 2016 was retrospectively queried. Patients with recurrent critical ischemia (Rutherford 4 and 5) were identified. Outcomes in this cohort were analyzed, and three groups were defined: endovascular reintervention (ie, a repeated tibial or pedal endovascular intervention), bypass (bypass to a tibial or pedal vessel), and primary amputation (ie, above- or below-knee amputation) on the ipsilateral leg. Patient-oriented outcomes of clinical efficacy (absence of recurrent signs or symptoms of critical ischemia, maintenance of ambulation, and absence of major amputation), amputation-free survival (survival without major amputation), and freedom from major adverse limb events (above-ankle amputation of the index limb or major reintervention, such as new bypass graft or jump or interposition graft revision) were evaluated after the reintervention. There were 1134 patients (56% male; average age, 59 years) who underwent primary tibial intervention for critical ischemia, and 54% presented with symptomatic restenosis and occlusion. Of the 513 patients with recurrent disease, 58% presented with rest pain and the remainder with ulceration. A repeated tibial endovascular intervention was performed in 64%, open bypass in 19%, and below-knee amputation in 17%. Bypass was employed in patients with a good target vessel, venous conduit, and good pedal runoff. Patient-centered outcomes were better in the bypass group compared with the reintervention group (amputation-free survival, 45% ± 9% vs 27% ± 9% [P < .01]; major adverse limb events, 50% ± 9% vs 31% ± 9% [P < .05]; clinical efficacy, 60% ± 7% vs 30% ± 9% [P < .01], mean ± standard error of the mean at 5 years). Tibial interventions for critical ischemia are associated with a high rate of reintervention. In patients with good target vessel, venous conduit, and good pedal runoff, bypass appears more durable than repeated tibial endovascular intervention.


Selection of characters for an Adansonian analysis of mycobacterial taxonomy.
Tests which are most frequently employed to provide characters for numerical analysis of bacterial taxonomy provide little differentiation among the slow-growing members of the genus Mycobacterium. A special scheme was proposed to deal with this problem. Characters were based, to some extent, on quantitative reactions to tests, and were defined, for each test, in terms of natural clustering behavior of a large collection of assorted strains. Inter- and intrataxon analyses were based on comparisons of reaction patterns with tentative hypothetical median strain patterns. This permitted the analysis of larger numbers of cultures than can be practically studied in a conventional n x n table.


Pre-treatment with Irbesartan attenuates left atrial stunning after electrical cardioversion of atrial fibrillation.
Left atrial (LA) stunning, the transient impairment of LA function, is responsible for an increased thrombo-embolic risk after cardioversion of atrial fibrillation (AF). Angiotensin receptor blockers (ARBs) attenuate atrial remodelling in AF and could theoretically influence LA stunning. We studied the effect of Irbesartan on LA stunning. We prospectively assigned 50 patients from the outpatient clinic undergoing electrical cardioversion for AF with duration of >4 weeks, into two matched groups: 25 patients were treated with Irbesartan (228+/-93 mg/day) for at least 2 weeks prior to cardioversion (Irbesartan group); 25 patients did not receive ARBs (control group). The groups did not differ concerning age (64+/-13 vs. 63+/-13 years, respectively), AF duration (20+/-18 vs. 20+/-19 weeks), underlying disease, LA diameter (46+/-7 vs. 47+/-9 mm), left ventricular dimensions, and ejection fraction (47.7+/-11.6 vs. 49.7+/-14.5%). We assessed LA appendage emptying velocities (LAAEV) and LA spontaneous echo contrast (LASEC) by transoesophageal echocardiography before and after cardioversion and at 2 weeks, and the A-wave by transthoracic echocardiography after cardioversion, at 2 and at 4 weeks. LA stunning was significantly attenuated in the Irbesartan group. The reduction of LAAEV immediately after cardioversion was significantly less in the Irbesartan group (LAAEV reduction of 9+/-49% from 28+/-9 cm/s before cardioversion to 25+/-13 cm/s immediately afterwards) than in the control group (reduction of 48+/-20% from 34+/-15 cm/s before cardioversion to 16+/-6 cm/s afterwards) (P = 0.048). New or increased LASEC occurred in eight patients (32%) in the Irbesartan vs. 16 patients (64%) in the control group (P = 0.046). Irbesartan significantly attenuates LA stunning after electrical cardioversion of AF. Therefore, ARBs may represent an important pharmacological supplementation in patients being prepared for cardioversion.


[Seasonal differences in the sensitivity of the catecholamine receptors of the brain in connection with the endocrine function of the pituitary-testicular complex].
The fluorimetric method was applied to the study of testosterone level in the peripheral blood plasma after the administration of substances stimulating catecholamine receptors of sexually mature Wistar male rats at different seasons of the year. Dophamine administration (a dose of 10 microgram) caused an increase in the blood testosterone level (twice) both in summer and in winter. At the same time noradrenaline stimulating chiefly the alpha-adrenoceptors and isopropylnoradrenaline (novodrin) acting chiefly on beta-adrenoceptors produced a stimulating action in the same doses in summer only. In winter the activation of the hypothalamo hypophysio-testicular complex was noted after the administration of these substances into the lateral ventricle in a dose of 100 microgram only. A conclusion was drawn that, in difference from dophamine receptors, sensitivity of alpha- and beta-adrenoceptors connected with the endocrine function of the hypothalamo-hypophysio-testicular complex depended on the seasonal factors. It is supposed that the brain adrenoceptors were capable of participating in the regulation of seasonal dynamics of testosterone content in the blood.


TBP2 gene may not be associated with primary ovarian insufficiency.
It has previously been reported that TATA-binding protein 2 (TBP2) knockout female mice are sterile. Our objective was to assess the association between the TBP2 gene and primary ovarian insufficiency (POI) in a Chinese population. A cohort of 60 POI patients matched with 60 fertile controls was recruited by Nanjing Medical University. There were no interventions. The complete TBP2 exon regions were analyzed by direct sequencing in all POI patients and controls. A known single nucleotide polymorphism (rs8019270) was identified in both POI and control groups. There was no difference in the genotype distribution or allelic frequencies between the control group and the POI group. No plausible pathogenic mutations were identified. Our study indicates that the TBP2 gene is not responsible for idiopathic POI in the Chinese Han population.


A new approach for resolving conflicts in actionable behavioral rules.
Knowledge is considered actionable if users can take direct actions based on such knowledge to their advantage. Among the most important and distinctive actionable knowledge are actionable behavioral rules that can directly and explicitly suggest specific actions to take to influence (restrain or encourage) the behavior in the users' best interest. However, in mining such rules, it often occurs that different rules may suggest the same actions with different expected utilities, which we call conflicting rules. To resolve the conflicts, a previous valid method was proposed. However, inconsistency of the measure for rule evaluating may hinder its performance. To overcome this problem, we develop a new method that utilizes rule ranking procedure as the basis for selecting the rule with the highest utility prediction accuracy. More specifically, we propose an integrative measure, which combines the measures of the support and antecedent length, to evaluate the utility prediction accuracies of conflicting rules. We also introduce a tunable weight parameter to allow the flexibility of integration. We conduct several experiments to test our proposed approach and evaluate the sensitivity of the weight parameter. Empirical results indicate that our approach outperforms those from previous research.


Generation and Characterization of a Spike Glycoprotein Domain A-Specific Neutralizing Single-Chain Variable Fragment against Porcine Epidemic Diarrhea Virus.
The emergence of the genotype (G) 2 and re-emergence of the G1 porcine epidemic diarrhea virus (PEDV) has caused severe economic impacts in the past decade. Developments of efficient vaccines against new variants of PEDV have been challenging, not least because of the difficulties in eliciting mucosal and lactogenic immunity. A single-chain fragment variable (scFv) capable of efficient antigen recognition is an alternative to vaccination and treatment of a viral infection. In the present study, the variable regions of the light chain and the heavy chain of a G2b PEDV spike domain A (S1A)-specific neutralizing monoclonal antibody (mAb) were sequenced, constructed with a (G4S) x3 linker, and produced by a mammalian protein expression system. Our results demonstrated that the PEDV S1A domain scFv was able to bind to S proteins of both G1 and G2b PEDVs. Nevertheless, the scFv was only capable of neutralizing the homologous G2b PEDV but not the G1 PEDV. The binding ability of the G2b-specific neutralizing scFv was not able to predict the neutralizing ability toward heterologous PEDV. The anti-PEDV S1A scFv presented herein serves as a potential therapeutic candidate against the virulent G2b PEDV.


Disseminated infection by Scedosporium prolificans: an emerging fatality among haematology patients. Case report and review.
We describe a case of proven disseminated infection by Scedosporium prolificans in a profoundly neutropenic patient. The patient presented with a fever unresponsive to broad-spectrum antibiotics, endophthalmitis, respiratory failure and a renal abscess. The organism was isolated from bronchoalveolar lavage fluid and from pus obtained through a sterile puncture. Review of the English-language literature identified 28 additional cases; these occurred exclusively in severely neutropenic patients (predominantly leukaemia) and in transplant recipients. Apart from two or possibly three cases, dissemination was uniformly fatal due to persistent neutropenia and inherited resistance of these pathogens to currently available antifungal drugs. At present, the optimal treatment of S. prolificans infections is unknown, but reversal of the underlying deficient immune status appears of great importance.


Cerebral-cortex hexokinase. Comparison of properties of solubilized mitochondrial and cytoplasmic activities.
1. Cerebral-cortex mitochondria, after purification by using high-density sucrose solutions, were extracted with Triton X-100. The total hexokinase activity of the intact mitochondria was increased by 50-80% in the Triton extracts. 2. Triton X-100 was removed from mitochondrial extracts by a combination of ammonium sulphate fractionation and DEAE-cellulose chromatography. Mitochondrial hexokinase remained soluble after removal of extractant. 3. The behaviour of solubilized mitochondrial hexokinase was compared with soluble cytoplasmic hexokinase from the same samples of cerebral cortex on identical columns of DEAE-cellulose. Two peaks were eluted from each source of hexokinase. The distribution between hexokinase peaks was similar for the two sources. Peak I (approx. 80% of the total hexokinase) from each was eluted at identical concentrations of potassium chloride and slight differences were observed in the elution profiles for peak II. 4. The purified mitochondrial hexokinase showed the following kinetic properties: peak I, K(m)(ATP) 0.60mm, K(m)(glucose) 0.042mm; peak II, K(m)(ATP) 0.66mm, K(m)(glucose) 0.043mm. The purified cytoplasmic hexokinase Michaelis constants were: peak I, K(m)(ATP) 0.56mm, K(m)(glucose) 0.048mm; peak II, K(m)(ATP) 0.68mm, K(m)(glucose) 0.062mm. 5. Although no significant differences between mitochondrial and cytoplasmic hexokinases were noted in chromatographic behaviour or in the kinetic properties studied, the purified mitochondrial enzyme was activated slightly (approx. 20%) by Triton X-100, in contrast with the cytoplasmic enzyme, which was not affected. 6. The results, taken to indicate basic similarity between mitochondrial and cytoplasmic hexokinases, are discussed in relation to the role of the two sources of enzyme in the metabolism of the tissue.


[Results of immunogenetic typing of lymphocytes of patients with keratoconus].
Histotyping of HLA I antigens in 100 patients with keratoconus. Europeoids of the Chelyabinsk district, and of 701 controls (donors listed in the regional register) showed an increased incidence of HLA A28, B12, and B15, a decreased incidence of HLA B8, B13, and complete absence of HLA A11. Estimation of HLA haplotypes in keratoconus showed an increased incidence of haplotypes A1-B5 (relative risk RR = 5.9), A2-B15 (RR = 3.34), A2-B27 (RR = 2.12), and A9-B21 (RR = 4.13) and decreased incidence of A1-B8 (RR = 0.31). The incidence of incomplete phenotypes was 27%. The most incident of incomplete phenotypes was A3-B35. Reliably linked HLA haplotypes A3-B7, A2-B27, A9-B21, and A9-B35 were typical of men with keratoconus, whereas A2-B21 haplotype was characteristic of the female patients.


Translation and validation of the Copenhagen Burnout Inventory amongst Greek doctors.
The Copenhagen Burnout Inventory (CBI) is a tool for burnout measurement and includes three different subscales: (i) personal, (ii) work-related and (iii) client-related burnout. The aim of this study was the translation and validation of CBI into the Greek Language. The forward-backward translation method was performed. Initially the questionnaire was distributed to 35 medical students for face validity assessment. Then, 284 residents answered the Greek version of CBI so as for construct validity to be examined. The data analysis was performed by using SPSS and AMOS. Face validity was estimated above 0.8 for the Greek version of CBI. The three factors model of CBI-Greek, with 1 item removed, achieved not so well-defined inner structure in CFI. However, the 16-item model achieved good levels of goodness-of-fit indices (Cmin/df 2.52, RMSEA 0.074, GFI 0.901, CFI70.938, NFI 0.901, TLI 0.923). The composite reliability values, also varied from 0.842 to 0.852. Cronbach's alpha index values surpassed 0.844. The 16-item model of CBI-Gr is a valid tool with high internal consistency that can be used in the evaluation of burnout in Greek population. The original 3-items model is a possible alternative.


Mesoaccumbal glutamate neurons drive reward via glutamate release but aversion via dopamine co-release.
Ventral tegmental area (VTA) projections to the nucleus accumbens (NAc) drive reward-related motivation. Although dopamine neurons are predominant, a substantial glutamatergic projection is also present, and a subset of these co-release both dopamine and glutamate. Optogenetic stimulation of VTA glutamate neurons not only supports self-stimulation but can also induce avoidance behavior, even in the same assay. Here, we parsed the selective contribution of glutamate or dopamine co-release from VTA glutamate neurons to reinforcement and avoidance. We expressed channelrhodopsin-2 (ChR2) in mouse VTA glutamate neurons in combination with CRISPR-Cas9 to disrupt either the gene encoding vesicular glutamate transporter 2 (VGLUT2) or tyrosine hydroxylase (Th). Selective disruption of VGLUT2 abolished optogenetic self-stimulation but left real-time place avoidance intact, whereas CRISPR-Cas9 deletion of Th preserved self-stimulation but abolished place avoidance. Our results demonstrate that glutamate release from VTA glutamate neurons is positively reinforcing but that dopamine release from VTA glutamate neurons can induce avoidance behavior.


[Microsatellite marker analysis of a soybean recombinant inbred line NJRIKY].
The prerequisite of genomic research such as genetic mapping is construction of a suitable segregating population and its reasonable evaluation before use. A recombinant inbred line (RILs), NJRIKY, was constructed through a cross between Kefeng 1 and Nannong 1138-2. The present study was focused on evaluation of genetic structure of the RILs through the simple sequence repeats (SSRs) analysis. A total number of 138 pairs of SSRs primers covering the whole soybean genome were randomly selected for polymorphic analysis between two parents. 86 pairs of which were polymorphic and polymorphic rate is up to 62.32%. 90 polymorphic loci were found. Further analysis with polymorphic SSRs loci on the sampled lines indicated that not only segregation of most SSRs loci conformed to the ratio of 1:1, but also almost each sampled line tended to homozyge. The distribution of genotypic composition was normal. The population is ideal for genetic mapping and other genomic research.


The association of remote Streptococcus bovis bacteremia with colonic neoplasia.
We prospectively studied five patients with a remote history of Streptococcus bovis bacteremia who had not previously been evaluated for colonic pathology. This study was prompted by several reports of concomitant neoplasia in patients with recent S. bovis bacteremia. The patients were studied at a mean of 32 months after bacteremia. Despite the absence of other signs or symptoms suggestive of bowel disease, a negative test for occult blood in stool and a negative fecal culture for S. bovis, two patients were found to have neoplastic polyps of the colon. These findings suggest that the risk of colonic neoplasia in patients with remote S. bovis septicemia is similar to that found in patients with recent bacteremia, but evaluation of a larger group of these patients will be necessary to establish this risk.


Clinical Benefits of Preoperative Conventional Fluorescence Diagnosis in Surgical Treatment of Extramammary Paget Disease.
In extramammary Paget disease (EMPD), initial margins of surgical excision are difficult to determine preoperatively. Fluorescence diagnosis (FD) may have utility as a noninvasive method for delineating tumor boundaries. The relationship between FD-delineated borders and results of multiple scouting biopsies (MSBs) in EMPD was investigated, evaluating the potential clinical role of FD. A cohort of 21 patients was studied, looking for consistencies between FD and MSB results. Initial surgical margins were determined by FD or FD plus MSB. All margins were subjected to frozen section analysis. Paget cells were identified in 88 of 117 (71.8%) biopsies obtained from FD borders. When used preoperatively with MSB in 9 patients, 96.4% of margins proved negative at initial resection stage. Using a 6-mm margin beyond FD borders in another 9 patients, 86.7% of margins were negative at initial stage. The maximum distance of excision beyond FD border was 12 mm. Fluorescence diagnosis borders (identified visually) correlated well with MSB histopathology. Fluorescence diagnosis is a useful method to delineate initial margins for surgical resection of EMPD.


Breast cancers with high proliferation and low ER-related signalling have poor prognosis and unique molecular features with implications for therapy.
Luminal breast cancers with high proliferation (MKShi) and low ER-related signalling (ERSlo) have a poor prognosis. We investigated treatment responses and molecular features of MKShi/ERSlo tumours to inform potential therapies. Gene expression data from patients who received neoadjuvant chemotherapy (NAC) without (MDACC, N = 199) or with pembrolizumab (I-SPY2, N = 40), or endocrine therapy (NET) without (POETIC, N = 172) or with palbociclib (NeoPalAna, N = 32) were analyzed to assess treatment response by MKS/ERS-subgroups. TCGA was used to assess the mutational landscape and biomarkers associated with palbociclib-resistance (Cyclin-E, RBsig, IRPR) and immunotherapy-response (TMB, TILs, T-cell inflamed) by MKS/ERS-subgroups. Compared to MKShi/ERShi tumours, MKShi/ERSlo tumours had higher pathological response rates to NAC (22% vs 8%, p = 0.06) but a higher recurrence risk (4-year metastasis-free survival 70% vs 94%, p = 0.01). MKShi/ERSlo tumours frequently harboured TP53 (34%) and PIK3CA (33%) mutations, and showed high expression of Cyclin-E, RBsig and IRPR, high TMB and elevated TIL and T-cell inflamed metagene expression. MKShi/ERSlo tumours retained high proliferation after NET with or without palbociclib but had higher pathological complete response rates when pembrolizumab was added to NAC (42% vs 21%, p = 0.07). MKShi/ERSlo tumours have dismal outcomes and are enriched in chemotherapy-sensitive but ET- and palbociclib-resistant tumours. Biomarker analysis and clinical data suggest a potential role for immunotherapy in this group.


Ligand-induced conformational change of Plasmodium falciparum AMA1 detected using 19F NMR.
We established an efficient means of probing ligand-induced conformational change in the malaria drug target AMA1 using 19F NMR. AMA1 was labeled with 5-fluorotryptophan (5F-Trp), and the resulting 5F-Trp resonances were assigned by mutagenesis of the native Trp residues. By introducing additional Trp residues at strategic sites within a ligand-responsive loop, we detected distinct conformational consequences when various peptide and small-molecule ligands bound AMA1. Our results demonstrate an increase in flexibility in this loop caused by the native ligand, as inferred from, but not directly observed in, crystal structures. In addition, we found evidence for long-range allosteric changes in AMA1 that are not observed crystallographically. This method will be valuable in ongoing efforts to identify and characterize therapeutically relevant inhibitors of protein-protein interactions involving AMA1 and is generalizable to the study of ligand-induced conformational change in a wide range of other drug targets.


Multifactorial determination of skeletal age at death: a method and blind tests of its accuracy.
Traditional methods of estimating skeletal age at death have relied solely on the pubic symphyseal face or on this indicator combined with others in nonsystematic ways. A multifactorial method is presented that uses a principal components weighting of five indicators (public symphyseal face, auricular surface, radiographs of proximal femur, dental wear, and suture closure). This method has been tested by completely blind assessment of age in two samples from the Todd collection carefully screened for accuracy of stated age at death. Results show a marked superiority of the multifactorial method over any single indicator with respect to both bias and accuracy. This represents the first truly blind test of an age-at-death indicator or system, as the test populations were independent of the system(s) being tested, and the age, sex, and ethnogeographic origin of the individuals being assessed (as well as the compositions of the test samples with respect to these variables) were completely unknown until the tests were completed. Implications for paleodemography are discussed.


Dynamic visualization of signaling activities in living cells.
The complexity and specificity of many forms of signal transduction are widely suspected to require spatial microcompartmentation and dynamic modulation of the activities of protein kinases, phosphatases, and second messengers. However, traditional methodologies for detecting signaling events, such as activation of kinases and second-messenger production and degradation, are limited in their spatiotemporal resolution and do not allow one to follow these events within the live-cell context. To achieve dynamic tracking of signaling activities in living cells, we have engineered genetically encoded fluorescent reporters for protein kinases and second messengers, such as cyclic adenosine monophosphate (cAMP) and phosphoinositides. Their development and specific examples of their application are discussed. In addition, a live-cell, high-throughput screening method has been developed for identification of new modulators that affect the dynamic activity of kinases and second messengers. Together, these reporters have the potential to provide important spatiotemporal information about the circuitry governing specific signaling events in living cells.


Grammar originates in action planning, not in cognitive and sensorimotor visual systems.
While the PREDICATE(x) structure requires close coordination of subject and predicate, both represented in consciousness, the cognitive (ventral), and sensorimotor (dorsal) pathways operate in parallel. Sensorimotor information is unconscious and can contradict cognitive spatial information. A more likely origin of linguistic grammar lies in the mammalian action planning process. Neurological machinery evolved for planning of action sequences becomes applied to planning communicatory sequences.


Sodium Orthovanadate and Trigonella Foenum Graecum Prevents Neuronal Parameters Decline and Impaired Glucose Homeostasis in Alloxan Diabetic Rats.
Hyperglycemia is the most important contributor in the onset and progress of diabetic complications mainly by producing oxidative stress. The present study was carried out to observe, the antihyperglycemic effect of sodium orthovanadate (SOV) and Trigonella foenum graecum seed powder (TSP) administration on blood glucose and insulin levels, membrane linked enzymes (monoamine oxidase, acetylcholinesterase, Ca2+ATPase), intracellular calcium (Ca2+) levels, lipid peroxidation, membrane fluidity and neurolipofuscin accumulation in brain of the alloxan induced diabetic rats and to see whether the treatment with SOV and TSP was capable of reversing the diabetic effects. Diabetes was induced by administration of alloxan monohydrate (15 mg/100 g body weight) and rats were treated with 2 IU insulin, 0.6 mg/ml SOV, 5% TSP in the diet and a combination of 0.2 mg/ml SOV and 5% TSP separately for three weeks. Diabetic rats showed hyperglycemia with almost four fold high blood glucose levels. Activities of acetylcholinesterase and Ca2+ATPase decreased in diabetic rat brain. Diabetic rats exhibited an increased level of intracellular Ca2+ levels, lipid peroxidation, neurolipofuscin accumulations and monoamine oxidase activity. Treatment of diabetic rats with insulin, TSP, SOV and a combined therapy of lower dose of SOV with TSP revived normoglycemia and restored the altered level of membrane bound enzymes, lipid peroxidation and neurolipofuscin accumulation. Our results showed that lower doses of SOV (0.2 mg/ml) could be used in combination with TSP in normalization of altered metabolic parameters and membrane linked enzymes without any harmful side effect.


[Significance of the involvement of parapharyngeal space cervical nodes and cranial nerves in nasopharyngeal carcinoma].
Two hundred cases of nasopharyngeal carcinoma (NPC) admitted to this department from Feb. 1985 to May. 1988 were analysed according to the CT scanning and clinical findings of the primary lesions prior to radiotherapy. The results showed that involvement of parapharyngeal space was very common in NPC, about 80% (160/200 cases); particularly unilateral or bilateral retro-styloid spaces, about 69.5% (139/200 cases). It was proposed that patients with NPC had a high incidence of ipsilateral cervical node metastasis. Contralateral cervical node metastasis was rare. The development of cervical node metastasis in NPC has to modes: one is direct infiltration of the retro-styloid space by the lesion; the other is along the nasopharyngeal lymphatic rete. The data also showed that patients with NPC who presented symptoms of IX-XII cranial nerve paralyses always had ipsilateral or bilateral retro-styloid space infiltrations.


Computational fragment-based drug design to explore the hydrophobic sub-pocket of the mitotic kinesin Eg5 allosteric binding site.
Eg5, a mitotic kinesin exclusively involved in the formation and function of the mitotic spindle has attracted interest as an anticancer drug target. Eg5 is co-crystallized with several inhibitors bound to its allosteric binding pocket. Each of these occupies a pocket formed by loop 5/helix alpha2 (L5/alpha2). Recently designed inhibitors additionally occupy a hydrophobic pocket of this site. The goal of the present study was to explore this hydrophobic pocket with our MED-SuMo fragment-based protocol, and thus discover novel chemical structures that might bind as inhibitors. The MED-SuMo software is able to compare and superimpose similar interaction surfaces upon the whole protein data bank (PDB). In a fragment-based protocol, MED-SuMo retrieves MED-Portions that encode protein-fragment binding sites and are derived from cross-mining protein-ligand structures with libraries of small molecules. Furthermore we have excluded intra-family MED-Portions derived from Eg5 ligands that occupy the hydrophobic pocket and predicted new potential ligands by hybridization that would fill simultaneously both pockets. Some of the latter having original scaffolds and substituents in the hydrophobic pocket are identified in libraries of synthetically accessible molecules by the MED-Search software.


Genetic diversity in natural populations of noble crayfish (Astacus astacus L.) in north-western Poland on the basis of combined SSR and AFLP data.
Conservation of noble crayfish (Astacus astacus) populations is becoming particularly important since the number of individuals is rapidly declining across the distribution range of the species in Europe. Five crayfish populations in northwestern Poland have been constantly monitored for two decades. However, the genetic structure of these populations has not been analysed, although this information is important to devise effective conservation strategies. Noble crayfish were collected in the autumn of 2014 by scuba diving in Lakes Graniczne, Babinki, Biwakowe, Sęki and Kwisno, all of which are situated in the Bytów Lakeland of northwestern Poland. Genetic diversity of the five populations was assessed based on allele variability in nine SSR regions and six AFLP primer combinations. Microsatellite results analysed with AMOVA showed that the diversity between populations corresponds to 18% of total variability, which was confirmed by similar results obtained using AFLP. Additionally, significant genetic diversity was revealed by high average FST values. All of the studied crayfish populations significantly deviated from the expected Hardy-Weinberg genetic equilibrium and were characterised by negative values of inbreeding coefficient (FIS). The invariably negative inbreeding coefficients (FIS) suggest a low number of mating individuals, a possible consequence of the phenomenon known as genetic bottleneck. However, additional comprehensive analyses are needed to assess the genetic structure, origin and vulnerability of the remaining populations of noble crayfish in the Bytów Lakeland of northwestern Poland, which have high conservation value and are particularly important as a live genetic bank for breeding and restitution purposes.


Effect of cholinergic agonists on bulbospinal C1 neurons in rats.
Cholinergic inputs to the rostral ventrolateral medulla (RVLM) may contribute to sympathetic tone generation. The present study analyzes the response of RVLM neurons to cholinergic agonists. In chloralose-anesthetized rats iontophoresis of carbachol excited RVLM sympathoexcitatory neurons (+69% from resting level of 11.9 +/- 2 spikes/s; n = 28). This effect was reduced 85% by iontophoresis of methylatropine and abolished by intravenous scopolamine. Iontophoresis of nicotine or hexamethonium was ineffective. In contrast, most RVLM respiratory units were inhibited by carbachol. Whole cell recordings of bulbospinal RVLM neurons were made in neonatal rat brain slices (54 cells, 24 C1 adrenergic neurons). In current-clamp recordings (without tetrodotoxin) carbachol produced depolarization, increased postsynaptic potential frequency, and decreased input resistance. In voltage-clamp recording (-50 to -60 mV; 1 microM tetrodotoxin) carbachol produced inward current [50% effective concentration (EC50): 10 +/- 1 microM; 12.6 +/- 2 pA at 30 microM; n = 16] that persisted in low Ca2+/high Mg2+ (n = 6). Muscarine (30 microM) caused smaller inward currents (2.6 +/- 0.6 pA; n = 16). The carbachol-induced current was reduced 46% by 5 microM methylatropine (n = 15) and 84% by 200 microM hexamethonium (n = 9). The current was linear as a function of the holding potential (extrapolated reversal potential: -22 +/- 2 mV). In conclusion, carbachol exerts both pre- and postsynaptic effects on C1 and other putative sympathoexcitatory RVLM neurons. In vitro the postsynaptic effect of carbachol has a mixed nicotinic and muscarinic pharmacology. In vivo, iontophoretically applied carbachol produces muscarinic excitation of barosensitive RVLM neurons.


The reduction of cystine during its transpot into rat jejunal eveted segments and sacs.
1. Everted segments and sacs of rat jejunum were incubated in buffer containing [(35)S]cystine. 2. Concentration gradients were achieved by both segments and sacs, and the effects of duration of incubation and of cystine concentration on the isotope distribution ratios were determined. 3. Kinetic constants were determined for the uptake of cystine by both segments and sacs, and the differences between the two systems are discussed. 4. Reduction to cysteine was virtually complete intracellularly and in the sac lumen. Extensive reduction in the medium occurred only when segments were incubated. 5. Anaerobiosis prevented a concentration gradient being obtained between the medium and the tissue, but had little effect on the extent of reduction to cysteine in the tissue and sac lumen. 6. It is concluded that cystine is transported by an active process into rat jejunum, where it is present almost entirely in the reduced form, and that efflux of cysteine occurs through the serosal surface.


Distribution of aluminium in hydrated leaves of tea (Camellia sinensis) using synchrotron- and laboratory-based X-ray fluorescence microscopy.
Aluminium (Al) is highly toxic to plant growth, with soluble concentrations being elevated in the ∼40% of arable soils worldwide that are acidic. Determining the distribution of Al in plant tissues is important for understanding the mechanisms by which it is toxic and how some plants tolerate high concentrations. Synchrotron- and laboratory-based X-ray fluorescence microscopy (XFM) is a powerful technique to quantitatively analyse the distribution of elements, including in hydrated and living plants. However, analysis of light elements (z < phosphorus) is extremely challenging due to signal losses in air, and the unsuitability of vacuum environments for (fresh) hydrated plant tissues. This study uses XFM in a helium environment to avoid Al signal loss to reveal the distribution of Al in hydrated plant tissues of Tea (Camellia sinensis). The results show that Al occurs in localised areas across the foliar surface, whereas in cross-sections Al is almost exclusively concentrated in the apoplastic space above and in between adaxial epidermal cells. This distribution of Al is related to the Al tolerance of this species, and accumulation of phytotoxic elements in the apoplastic space, away from sensitive processes such as photosynthesis in the palisade mesophyll cells, is a common tolerance mechanism reported in many different plant species. This study develops an XFM method on both synchrotron and laboratory sources that overcomes the drawbacks of existing analytical techniques, permitting measurement of light elements down to Al in (fresh) hydrated plant tissues.


Sonochemical synthesis of Eu3+ substituted CoFe2O4 nanoparticles and their structural, optical and magnetic properties.
Magnetic, optic and microstructural properties of ultrasonically synthesized CoEuxFe2-xO4 (x ≤ 0.1) nanoferrites (NFs) have been examined in this study. After sonochemical synthesis, XRD and FT-IR analyses confirmed the purity, the structure (cubic spinel structure and Fd3m space group) and the spectral properties of the spinel ferrite samples. The spherical morphology and chemical compositions of the products were observed via transmission and scanning electron microscopes along with EDX and elemental mapping. Percent diffuse reflectance (%DR) was used for optical investigation. Optical band gaps (Eg) were estimated utilizing Kubelka-Munk theory and Tauc equation. Eg values are in a narrow band of 1.34 to 1.44 eV. The magnetic parameters like Ms (saturation magnetization), SQR = Mr/Ms (squareness ratio), nB (magnetic moment), Hc (coercivity) and Mr (remanence) have been evaluated by analyzing measurements of magnetization versus magnetic field performed at room (RT; T = 300 K) and low (T = 10 K) temperatures. It is showed that the different produced CoEuxFe2-xO4 (0.00 ≤ x ≤ 0.10) nanospinel ferrites present superparamagnetic (SPM) nature at RT. At low temperature, the various produced CoEuxFe2-xO4 (x ≤ 0.08) nanospinel ferrites display ferrimagnetic (FM) nature. With exception, the x = 0.10 sample exhibit SPM behavior at T = 10 K. It is noticed that the Eu3+ substitutions alter in a significant way on the magnetic data. A decreasing trend in the Ms, Mr and nB values was noted with Eu3+ substitutions.


Occluded but nonthrombosed internal carotid artery: an indication for endarterectomy.
The indications for carotid endarterectomy are sometimes imprecise. Total occlusion of the internal carotid artery has often implied irreversibility. Carotid thromboendarterectomy was performed on eight patients who on angiography had a complete occlusion of proximal internal carotid artery but showed antegrade opacification of the infraophthalmic carotid siphon. Direct magnification, delayed radiography, and subtraction were used on all patients. In five of eight patients, normal antegrade cervical internal carotid blood flow was restored. The possible restoration of the occluded internal carotid artery lumen offers a viable alternative to bypass surgery in selected cases.


Predictors of cholesterol and lipoprotein(a) testing in children with arterial ischemic stroke.
Professional societies recommend screening lipids in healthy children. Dyslipidemia and elevated lipoprotein(a) are risk factors for adult cardiovascular disease and stroke. Their role in childhood arterial ischemic stroke is unexplored. Inconsistencies in testing limit analysis of existing lipid data. The objective of this study is to identify predictors and modifiable barriers to lipid testing in pediatric stroke. In this cross-sectional analysis, children (28 days-18 years) with arterial ischemic stroke were identified from the International Pediatric Stroke Study registry (January 2003-April 2012). Analyzed predictors of recorded lipid or lipoprotein a (Lp(a)) testing were age, sex, race, ethnicity, body mass index (BMI) category, other stroke risk factors, country, US region, and recurrent thrombosis. Among 1652 participants (median, 6 years [interquartile range, 1.7-12.7]; 59.0% male; 40.8% white; 7.0% black), at least 1 lipid parameter or Lp (a) was available for 461 (27.9%). Compared with infants, testing was incrementally higher for older age categories. Compared with whites, testing was lower in blacks (adjusted odds ratio [OR], .5; 95% confidence interval [CI], .4-.5; P < .0001). Hispanic ethnicity only predicted testing within the United States (OR, 2.2; 95% CI, 1.4-3.4; P = .001]. Testing was lower in the United States and Australia and higher in Chile. Any thrombotic recurrence and recurrent symptomatic arterial ischemic stroke were associated with testing, unlike male sex, BMI, other stroke risk factors, and region in the United States. Only a quarter of children with stroke had recorded lipid testing. Older age, white race, and recurrence predicted testing. In future study adjusting for these predictors may be necessary. Standardized lipid testing in children with arterial ischemic stroke may further our understanding of this potential risk factor.


Frequency and prognostic value of resistance/intolerance to hydroxycarbamide in 890 patients with polycythaemia vera.
The clinical significance of resistance/intolerance to hydroxycarbamide (HC) was assessed in a series of 890 patients with polycythaemia vera (PV). Resistance/intolerance to HC was recorded in 137 patients (15·4%), consisting of: need for phlebotomies (3·3%), uncontrolled myeloproliferation (1·6%), failure to reduce massive splenomegaly (0·8%), development of cytopenia at the lowest dose of HC to achieve a response (1·7%) and extra-haematological toxicity (9%). With a median follow-up of 4·6 years, 99 patients died, resulting in a median survival of 19 years. Fulfilling any of the resistance/intolerance criteria had no impact on survival but when the different criteria were individually assessed, an increased risk of death was observed in patients developing cytopenia [Hazard ratio (HR): 3·5, 95% confidence interval (CI): 1·5-8·3, P = 0·003]. Resistance/intolerance had no impact in the rate of thrombosis or bleeding. Risk of myelofibrotic transformation was significantly higher in those patients developing cytopenia (HR: 5·1, 95% CI: 1·9-13·7, P = 0·001) and massive splenomegaly (HR: 9·1, 95% CI: 2·3-35·9, P = 0·002). Cytopenia at the lowest dose required to achieve a response was also an independent risk factor for transformation to acute leukaemia (HR: 20·3, 95% CI: 5·4-76·5, P < 0·001). In conclusion, the unified definition of resistance/intolerance to HC delineates a heterogeneous group of PV patients, with those developing cytopenia being associated with an adverse outcome.