Equine infectious anemia: preparation of a liquid antigen extract for the agar-gel immunodiffusion and complement-fixation tests.
An agar-gel immunodiffusion test recommended for the diagnosis of equine infectious anemia was evaluated. Our preliminary observations confirmed those of Coggins concerning the mechanism of the test and the results obtained. Furthermore, emphasis was put on the difficulties encountered in the production of spleen antigens with an optimum amount of reactivity. Acetone-ether extraction procedures for the preparation of a liquid antigen extract are described. This type of antigen was reactive in the complement-fixation test in 1:8 or greater dilution and it is proposed to use the complement-fixation test in assessing and standardizing the liquid antigen extract activity to be used in the immunodiffusion test. This antigen can also be concentrated or diluted, if required, to meet the reactivity of a standard antigen used in the test.


The two largest chloroplast genome-encoded open reading frames of higher plants are essential genes.
The chloroplast genomes of most higher plants contain two giant open reading frames designated ycf1 and ycf2. In tobacco, ycf1 potentially specifies a protein of 1901 amino acids. The putative gene product of the ycf2 reading frame is a protein of 2280 amino acids. In an attempt to determine the functions of ycf1 and ycf2, we have constructed several mutant alleles for targeted disruption and/or deletion of these two reading frames. The mutant alleles were introduced into the tobacco plastid genome by biolistic chloroplast transformation to replace the corresponding wild-type alleles by homologous recombination. Chloroplast transformants were obtained for all constructs and tested for their homoplastomic state. We report here that all transformed lines remained heteroplastomic even after repeated cycles of regeneration under high selective pressure. A balanced selection was observed in the presence of the antibiotic spectinomycin, resulting in maintenance of a fairly constant ratio of wild-type versus transformed genome copies. Upon removal of the antibiotic and therewith release of the selective pressure, sorting out towards the wild-type plastid genome occurred in all transplastomic lines. These findings suggest that ycf1 and ycf2 are functional genes and encode products that are essential for cell survival. The two reading frames are thus the first higher plant chloroplast genes identified as being indispensable.


Analyses of delta(13)C and delta(18)O in tree rings of Callitris columellaris provide evidence of a change in stomatal control of photosynthesis in response to regional changes in climate.
We examined relationships between stable isotopes of carbon (delta(13)C) and oxygen (delta(18)O) in tree rings of Callitris columellaris F. Muell in the semi-arid Pilbara region of north-western Australia. To test the hypothesis that stomatal control of photosynthesis decreases during drier periods, we developed delta(13)C and delta(18)O chronologies spanning 1919-1999, and used a permutation regression approach to relate a 21-year running correlation between delta(13)C and delta(18)O to rainfall and temperature at Marble Bar and our study site. The relationship between delta(13)C and delta(18)O switched from being always negative before 1955 to being consistently positive after 1976, suggesting an increase in stomatal control of photosynthesis in recent decades. Changes in the delta(13)C-delta(18)O relationship reflected changes in rainfall, which has increased in the region by 30% since 1976. The correlation between delta(13)C and delta(18)O was positively related to the 21-year running mean of normalized rainfall anomalies at both the study site (P = 0.045, Adj. r(2) = 0.47) and Marble Bar (P = 0.046, Adj. r(2) = 0.48). In addition, the delta(13)C-delta(18)O correlation was negatively related (P = 0.047, Adj. r(2) = 0.61) to temperatures at Marble Bar. Our interpretation of the role of changes in climate affecting the relationship between tree-ring delta(13)C and delta(18)O is supported by evidence from the isotope composition of foliage samples: foliar delta(13)C and delta(18)O were negatively correlated with log stomatal conductance (delta(13)C, r = -0.41; delta(18)O, r = -0.42), whereas the correlation between foliar delta(13)C and delta(18)O was positive (r = 0.63, P = 0.027) after the summer wet period. Our data indicate that stomatal control of photosynthesis in Callitris adjusts to region-wide changes in climate and that, in a warmer and drier world, trees might adapt by increasing non-stomatal control of photosynthesis.


The Effects of Olive Leaf Extract on The Testis, Sperm Quality and Testicular Germ Cell Apoptosis in Male Rats Exposed to Busulfan.
Busulfan (BU) has a destructive effect on the male reproductive system. The goal of this study was to assess the effects of olive leaf extract (OLE) as a source of antioxidants and phenolic compounds, on BU-induced damages in rat testes. In this experimental study, 40 male Wistar rats were randomly divided into 5 groups. The control group (CTL) received a single intraperitoneal (i.p.) injection of dimethyl sulfoxide (DMSO), followed by oral administration of distilled water for 5 weeks. In BU group, BU (10 mg/kg) was administrated i.p. once. In cotreatment groups, first, received BU (10 mg/kg, a single i.p. injection) then, OLE was administrated orally at different doses of 250 mg/kg (BU+OLE 250), 500 mg/kg (BU+OLE 500) and 750 mg/kg (BU+OLE 750), for 5 weeks. Next, blood and sperm samples were collected. The left testis was removed to investigate testicular parameters and apoptosis by using H and E and TUNEL staining, respectively. All data were analyzed by SPSS software and a P<0.05 was considered significant. There was a significant decline in sperm viability (P=0.017), number of primary spermatocyte (PS) (P=0.001) and Leydig cells (P=0.023) in the BU group versus the CTL group. OLE at three doses could repair these defects versus BU group. Increases in apoptotic spermatogonia cells (SG) due to BU were significantly reduced by OLE 250 and 500 mg/kg (P<0.01). A reduction in germinal epithelium height and an increase in apoptotic SG were observed in BU+OLE 750 group vs. other groups (P<0.01) and alkaline phosphatase (ALP) was at the highest level, also Aspartate aminotransferase (AST) increased markedly vs. CTL (P=0.024). Oral administration of OLE at the doses of 250 and 500 mg/kg could be helpful in ameliorating BUinduced toxicity in rat testes, while OLE 750 mg/kg not only did not cause positive effects, but also could exacerbate the harmful effects.


Researcher perspectives on challenges and opportunities in conservation physiology revealed from an online survey.
Conservation physiology represents a recently emerging arm of conservation science that applies physiological tools and techniques to understand and solve conservation issues. While a multi-disciplinary toolbox can only help to address the global biodiversity crisis, any field can face challenges while becoming established, particularly highly applied disciplines that require multi-stakeholder involvement. Gaining first-hand knowledge of the challenges that conservation physiologists are facing can help characterize the current state of the field and build a better foundation for determining how it can grow. Through an online survey of 468 scientists working at the intersection of physiology and conservation, we aimed to identify characteristics of those engaging in conservation physiology research (e.g. demographics, primary taxa of study), gauge conservation physiology's role in contributing to on-the-ground conservation action, identify the perceived barriers to achieving success and determine how difficult any identified barriers are to overcome. Despite all participants having experience combining physiology and conservation, only one-third considered themselves to be 'conservation physiologists'. Moreover, there was a general perception that conservation physiology does not yet regularly lead to tangible conservation success. Respondents identified the recent conceptualization of the field and the broader issue of adequately translating science into management action as the primary reasons for these deficits. Other significant barriers that respondents have faced when integrating physiology and conservation science included a lack of funding, logistical constraints (e.g. sample sizes, obtaining permits) and a lack of physiological baseline data (i.e. reference ranges of a physiological metric's 'normal' or pre-environmental change levels). We identified 12 actions based on suggestions of survey participants that we anticipate will help deconstruct the barriers and continue to develop a narrative of physiology that is relevant to conservation science, policy and practice.


Idiopathic partial epilepsy with auditory features (IPEAF): a clinical and genetic study of 53 sporadic cases.
The purpose of our study was to describe the clinical characteristics of sporadic (S) cases of partial epilepsy with auditory features (PEAF) and pinpoint clinical, prognostic and genetic differences with respect to previously reported familial (F) cases of autosomal dominant partial epilepsy with auditory features (ADPEAF). We analysed 53 patients (24 females and 29 males) with PEAF diagnosed according to the following criteria: partial epilepsy with auditory symptoms, negative family history for epilepsy and absence of cerebral lesions on NMR study. All patients underwent a full clinical, neuroradiological and neurophysiological examination. Forty patients were screened for mutations in LGI1/epitempin, which is involved in ADPEAF. Age at onset ranged from 6 to 39 years (average 19 years). Secondarily generalized seizures were the most common type of seizures at onset (79%). Auditory auras occurred either in isolation (53%) or associated with visual, psychic or aphasic symptoms. Low seizure frequency at onset and good drug responsiveness were common, with 51% of patients seizure-free. Seizures tended to recur after drug withdrawal. Clinically, no major differences were found between S and F patients with respect to age at onset, seizure frequency and response to therapy. Analysis of LGI1/epitempin exons failed to disclose mutations. Our data support the existence of a peculiar form of non-lesional temporal lobe epilepsy closely related to ADPEAF but without a positive family history. This syndrome, here named IPEAF, has a benign course in the majority of patients and could be diagnosed by the presence of auditory aura. Although LGI1 mutations have been excluded, genetic factors may play an aetiopathogenetic role in at least some of these S cases.


Economics of vaccines revisited.
Performing a total health economic analysis of a vaccine newly introduced into the market today is a challenge when using the conventional cost-effectiveness analysis we normally apply on pharmaceutical products. There are many reasons for that, such as: the uncertainty in the total benefit (direct and indirect) to be measured in a population when using a cohort model; (1) appropriate rules about discounting the long-term impact of vaccines are absent jeopardizing therefore their value at the initial investment; (2) the presence of opposite contexts when introducing the vaccine in developed vs. the developing world with high benefits, low initial health care investment for the latter vs. marginal benefit and high cost for the former; with a corresponding paradox for the vaccine becoming very cost-effective in low income countries but rather medium in middle low to high middle income countries; (3) and the type of trial assessment for the newer vaccines is now often performed with immunogenicity reaction instead of clinical endpoints which still leaves questions on their real impact and their head-to-head comparison. (4.)


Understanding the Basics of Computational Models in Orthopaedics: A Nonnumeric Review for Surgeons.
Computational models represent more than just finite element analysis, a term that many clinicians may know and globally apply. Over the past 30 years, many published studies have addressed clinically relevant orthopaedic questions with speed and precision by using a wide variety of computational approaches. Given such a wide spectrum of techniques, clinicians often do not have a full understanding of the methods used to create models and therefore do not appreciate the strengths, weaknesses, and potential pitfalls of published results. The short, nonnumeric summaries of the methodologies employed for various computational approaches presented here can help address this issue.


The impact of drought on leaf physiology of Quercus suber L. trees: comparison of an extreme drought event with chronic rainfall reduction.
Understanding the responses of cork oak (Quercus suber L.) to actual and predicted summer conditions is essential to determine the future sustainability of cork oak woodlands in Iberia. Thermal imaging may provide a rapid method for monitoring the extent of stress. The ecophysiology of cork trees was studied over three years. Three treatments were applied by means of rainfall capture and irrigation, with plots receiving 120%, 100%, or 80% of natural precipitation. Despite stomatal closure, detected using both thermal imaging and porometry, leaf water potential fell during the summer, most drastically during the third year of accumulative stress. The quantum efficiency (ΦPSII) and the maximum efficiency Fv' /FM' of photosystem II also fell more intensely over the third summer, while non-photochemical quenching (NPQ) increased. The reduced precipitation treatment sporadically further reduced leaf water potential, stomatal conductance (gs), IG (an index of gs derived from thermal imaging), ΦPSII, and Fv' /FM', and increased leaf temperature and NPQ. It is concluded that these are very resilient trees since they were only severely affected in the third year of severe drought (the third year registering 45% less rainfall than average), and removing 20% of rainfall had a limited impact..


Conjugal transfer of antibiotic resistance factors in Bacteroides fragilis: the btgA and btgB genes of plasmid pBFTM10 are required for its transfer from Bacteroides fragilis and for its mobilization by IncP beta plasmid R751 in Escherichia coli.
Transferable plasmids play an important role in the dissemination of clindamycin-erythromycin resistance in Bacteroides fragilis. We previously described the isolation and properties of pBFTM10, a 14.9-kb ClnR transfer factor from B. fragilis TMP10. We also reported the isolation of a transfer-deficient deletion derivative of pBFTM10 contained in the B. fragilis-Escherichia coli shuttle vector pGAT400. In the present study we used pGAT400 and a similar shuttle vector, pGAT550, to characterize and sequence a region of pBFTM10 required for its transfer from B. fragilis to B. fragilis or E. coli recipients and for its mobilization by the broad-host-range plasmid R751 from E. coli donors to E. coli recipients. Deletion of certain BglII restriction fragments from pBFTM10 resulted in partial or complete loss of transfer ability. Tn1000 insertions into this same region also resulted in altered transfer properties. We used the sites of Tn1000 insertions to determine the DNA sequence of the transfer region. Two potential open reading frames encoding proteins of 23.2 and 33.8 kDa, corresponding to two genes, btgA or btgB, were identified in the sequence. Tn1000 insertions within btgA or btgB or deletion of all or portions of btgA or btgB resulted in either a transfer deficiency or greatly reduced transfer from B. fragilis donors and alterations in mobilization by R751 in E. coli. A potential oriT sequence showing similarity in organization to the oriT regions of the IncP plasmids was also detected. Thus, pBFTM10 encodes and requires at least two proteins necessary for efficient transfer from B. fragilis. These same functions are expressed in E. coli and are required for mobilization by R751.


Effects of sub-chronic CRH administration into the hypothalamic paraventricular and central amygdala nuclei in male rats with a focus on food intake biomarkers.
CRH neurons are found in the paraventricular nucleus(PVN) and central amygdala(CeA) nuclei. This study investigated the effects of sub-chronic CRH administration into the PVN and CeA nuclei on food intake biomarkers in rats divided into five groups: control, two shams, and two CRH-PVN and CRH-CeA groups(receiving CRH in nuclei for seven days). The CRH-PVN group had significantly higher cumulative food intake and food intake trends than the CRH-CeA group. The CRH-CeA and CRH-PVN groups exhibited significant increases in food intake during hours 1 and 2, respectively. Moreover, to be time-dependent, food intake is modulated by different brain nuclei. The CRH signaling pathway appeared to be activated later in the PVN than CeA. Both groups exhibited significantly higher leptin levels, the CRH-PVN group exhibited higher ghrelin levels and lower glucose levels. Repetitive administration of CRH into the PVN and CeA significantly reduced body weight differences. CRH administration into the PVN affected both leptin and ghrelin levels, but ghrelin had a greater impact on glucose variations and cumulative food intake than leptin. Finally, CRH administration into the PVN and CeA likely activated the HPA axis, and the CeA had a greater impact on the stress circuit than on food intake behavior.


Risk of cytomegalovirus infection and disease after umbilical cord blood transplantation in children.
Pediatric data regarding cytomegalovirus (CMV) infections in pediatric patients receiving umbilical cord blood (UCB) transplantation are sparse. To determine whether UCB transplantation increases the risk of CMV infection and disease compared with other graft sources. The medical files of patients who underwent allogeneic hematopoietic stem cell transplantation at CHU Ste-Justine (Montreal, Quebec) from April 2000 to December 2006 were retrospectively reviewed. A Cox proportional hazard model was used to assess the effect of potential predictors of outcomes. A total of 176 patients with a median age of nine years (range 0.1 to 18 years) underwent hematopoietic stem cell transplantation. The source of stem cells were UCB, bone marrow and peripheral blood stem cells in 86, 86 and four of the cases, respectively. CMV infection occurred in 29 patients (16%). At day 100 post-transplantation, the rate of CMV infection was 13% in UCB transplant recipients (11 of 86) versus 20% in those with other sources of graft (18 of 90) (P=0.19). Positive CMV serology of the recipient and leukocyte depletion were two independent variables associated with an increased risk of CMV infection. Among infected patients, six developed CMV disease (20.7%). The rate of CMV disease one year after infection was 49% in patients who received UCB (five of 11) and 6% in others (one of 18). This difference was significant by univariate (P=0.01) but not by multivariate analysis. In the setting of the current study, with a moderate CMV infection rate (16.5%), UCB transplantation did not appear to increase the risk of CMV infection and disease.


Comparison of two commercially available media for detection of bacteremia.
An analysis of 3,795 positive blood cultures obtained from 1,718 patients in a 2.5-year evaluation of Tryptic Soy Broth (TSB) and Thiol Broth is reported. Isolation rates of Actinobacillus and Pseudomonas were significantly greater in TSB, whereas isolation rates of Streptococcus and Corynebacterium (aerobic and anaerobic) were significantly greater in Thiol. Otherwise, the two media were similar. Disregarding contaminants, anaerobic bacteria represented 11% of positive cultures and 20% of patients with bacteremia. Eleven per cent of the patients had polymicrobial bacteremia.


Optical Coherence Tomography: Is Really a New Biomarker for Alzheimer's Disease?
Retinal ganglion cell (RGC) degeneration was histopathologically proved previously in Alzheimer's disease (AD) patients. In this study, we aimed to determine RGC degeneration in vivo using optical coherence tomography (OCT) in AD. Twenty-one mild-to-moderate AD patients and 25 cognitively healthy age-matched controls were enrolled in this case-control prospective study. All participants underwent OCT examination to assess peripapillary retinal nerve fiber layer (RNFL) thickness, macular volume, and thickness. Foveal thickness and volume were significantly higher in AD patients than controls (P = 0.023 and P = 0.024, respectively). Compared to controls, peripapillary RNFL and other macular region measurements of AD patients were not statistically different (for all P > 0.05). Increased foveal thickness and volume can be associated with the pathological changes in the early stages of degeneration These results differ from previous studies, but still confirm retinal degeneration in AD. With further OCT studies on large populations, OCT will be in clinical use for early diagnosis of AD.


Antinociceptive effects of the non-selective cannabinoid receptor agonist CP 55,940 are absent in CB1(-/-) and not CB2(-/-) mice in models of acute and persistent pain.
Previous studies have suggested a role for both CB1 and CB2 cannabinoid receptors in modulation of nociception. To further examine the role of CB1 and CB2 receptors in antinociception, we evaluated the efficacy of the non-selective cannabinoid receptor agonist, CP 55,940, in models of acute, inflammatory, and neuropathic pain in control mice, CB1 receptor knockout mice, and CB2 receptor knockout mice. In control C57BL/6 mice, administration of CP 55,940 (0.03-0.3 mg/kg, i.p.) reversed complete Freund's adjuvant-induced tactile allodynia, reversed tactile allodynia in the spinal nerve ligation model and inhibited the noxious heat-evoked tail withdrawal response. In addition to its antinociceptive effects, CP 55,940 produced an impairment of motor coordination in the rotarod test. The antinociceptive effects produced by CP 55,940 and associated motor deficits were found to be completely abolished in CB1 receptor knockout mice. In contrast, the antinociceptive effects of CP 55,940 in all pain models were fully retained in CB2 receptor knockout mice, along with the associated motor deficits. The results suggest that the antinociceptive effects of CP 55,940 in models of acute and persistent pain, along with the associated motor deficits, are mediated by CB1 receptors, and likely not CB2 receptors.


Understanding the HIV coreceptor switch from a dynamical perspective.
The entry of HIV into its target cells is facilitated by the prior binding to the cell surface molecule CD4 and a secondary coreceptor, mostly the chemokine receptors CCR5 or CXCR4. In early infection CCR5-using viruses (R5 viruses) are mostly dominant while a receptor switch towards CXCR4 occurs in about 50% of the infected individuals (X4 viruses) which is associated with a progression of the disease. There are many hypotheses regarding the underlying dynamics without yet a conclusive understanding. While it is difficult to isolate key factors in vivo we have developed a minimal in silico model based on the approaches of Nowak and May to investigate the conditions under which the receptor switch occurs. The model allows to investigate the evolution of viral strains within a probabilistic framework along the three stages of disease from primary and latent infection to the onset of AIDS with a a sudden increase in viral load which goes along with the impairment of the immune response. The model is specifically applied to investigate the evolution of the viral quasispecies in terms of R5 and X4 viruses which directly translates into the composition of viral load and consequently the question of the coreceptor switch. The model can explain the coreceptor switch as a result of a dynamical change in the underlying environmental conditions in the host. The emergence of X4 strains does not necessarily result in the dominance of X4 viruses in viral load which is more likely to occur in the model after some time of chronic infection. A better understanding of the conditions leading to the coreceptor switch is especially of interest as CCR5 blockers have recently been licensed as drugs which suppress R5 viruses but do not seem to necessarily induce a coreceptor switch.


Electronic spectroscopy of UO2(2+), NUO(+) and NUN: an evaluation of time-dependent density functional theory for actinides.
The performance of the time-dependent density functional theory (TDDFT) approach has been evaluated for the electronic spectrum of the UO(2)(2+), NUO(+) and NUN molecules. Different exchange-correlation functionals (LDA, PBE, BLYP, B3LYP, PBE0, M06, M06-L, M06-2X, CAM-B3LYP) and the SAOP model potential have been investigated, as has the relative importance of the adiabatic local density approximation (ALDA) to the exchange-correlation kernel. The vertical excitation energies have been compared with reference data obtained using accurate wave-function theory (WFT) methods.


Acute hemichorea in a newly diagnosed type II diabetes patient: a diagnostic challenge in resource-limited setting: a case report.
Chorea is a rare complication of uncontrolled type II diabetes. We report for the first time in Tanzania a case of type II diabetes presenting with a hyperglycaemia-induced hemichorea. A 58-year-old Tanzanian chagga by tribe with a body mass index of 28 kg/m(2) and newly diagnosed type II diabetes presented with polydipsia and involuntary movements of the right upper limb for 4 days. His plasma glucose was 549 mg/dl and glycated haemoglobin was 18.9 %. His movements were exaggerated by attempts to use his right hand. The rest of his neurological assessment was unremarkable. Other laboratory findings including calcium were within the normal range. A computed tomography scan of the brain was essentially normal except for age-related atrophy. There was no significant ketonuria on urine dipstick testing. We treated the patient's hyperglycaemia with intravenous insulin and the dystonia disappeared within 5 days. Hemichorea is among the rare complications of hyperglycaemia-induced involuntary movements. Hyperglycaemia should be considered as a differential diagnosis for patients with type II diabetes mellitus presenting with hemichorea upon clinical assessment.


[Prevalence of genetic diseases in admissions to a tertiary care hospital pediatric service].
With the epidemiological changes, the role of genetic factors as a cause of morbidity and mortality is increasing, changing disease patterns of patients admitted to pediatric hospitals. To describe the prevalence of genetic diseases (GD) in patients admitted to a tertiary-care hospital Pediatric Service. The clinical records of consecutive admissions to a Pediatric Service of a clinical hospital in 2011 were reviewed. Two categories were assigned: with GD and without GD. Both groups were compared according to days of hospitalization, type of admission, readmissions and mortality. We reviewed the 98.1% of the 1,781 total annual admissions (1,459 cases), 322 of them were readmissions (187 cases). The mean age at admission was 54.8 ± 54 months and 55% were male. The mean hospitalization length was 4.9 ± 10 days. Of total admissions and individual cases, 52.7% (938/1,781) and 48% (705/1,459) were cases with GD, respectively. Within this group, 85% (597/705) were sub-categorized as having a significant genetic base. The differences between gender, age average income and hospital mortality were not significant between the two categories. Readmissions were more common for GD than for patients without GD (Odds ratio (OR): 2.6, confidence intervals (CI): 1.9-3.6). Average hospital stay was 27% higher among GD patients (p < 0.01). Our findings confirm the high prevalence of GD in pediatric hospitals (52.7%), with a higher risk for readmission in cases with GD compared with those without GD.


Multivariate analysis applied to Raman mapping of dye-functionalized carbon nanotubes: a novel approach to support the rational design of functional nanostructures.
Principal component analysis is applied to analyse the Raman maps collected on carbon nanotubes at different degrees of oxidation and functionalization with dye labeling molecules. The results are used to demonstrate that the technique is extremely effective in clustering data and comparing preparation protocols, so that it enables drawing of a fast and reliable classification of the molecule propensity to interact with pristine and oxidized carbon nanotubes. The spectral findings are supported and elucidated by several experimental techniques, thermogravimetry and steady-state and time-resolved fluorescence measurements, and by computational modeling, showing that the proposed methodology could represent a powerful and routine test for the rational design of functional nanostructures.


The alien in the forest OR when temporal context dominates perception.
Some German hunters came up with pictures taken from a wildlife camera and showing a strange humanoid-like object of about 10 cm size. Its puzzling appearance and a remarkable absence of wildlife in that area after its occurrence triggered paranormal explanations. After reexamination of the pictures, we found a more plausible conventional explanation. This case study serves as an interesting real-world example for the constructive nature of human perception. We discuss how the perceptual system uses spatial and temporal/memory factors to disambiguate ambiguous and restricted sensory information.


Experience alters the spatial tuning of auditory units in the optic tectum during a sensitive period in the barn owl.
The auditory spatial tuning of bimodal (auditory-visual) units in the optic tectum of the barn owl was altered by raising animals with one ear occluded. Changes in spatial tuning were assessed by comparing the location of a unit's auditory best area with that of its visual receptive field. As shown previously, auditory best areas are aligned with visual receptive fields in the tecta of normal birds (Knudsen, E. I. (1982) J. Neurosci. 2: 1177-1194). It was demonstrated in this study that, when birds were raised with one ear occluded, best areas and visual receptive fields were aligned only as long as the earplug was in place. When the earplug was removed, best areas and visual receptive fields became misaligned, indicating that a change in auditory spatial tuning had taken place during the period of occlusion. However, in a bird that received an earplug as an adult, no such alterations in auditory spatial tuning were observed; even after 1 year of monaural occlusion, auditory best areas and visual receptive fields were misaligned so long as the earplug was in place, and were aligned when the earplug was removed. These results suggest that exposure to abnormal localization cues modifies the auditory spatial tuning of tectal units only during a restricted, sensitive period early in development. After the earplug was removed from a juvenile bird that had been raised with an occluded ear, the initial misalignment between auditory best areas and visual receptive fields decreased gradually over a period of weeks. In contrast, when earplugs were removed from two adult birds that had been raised with monaural occlusions, auditory-visual misalignments persisted for as long as measurements were made, which was up to 1 year after earplug removal. These data indicate that auditory cues become permanently associated with locations in visual space during a critical period which draws to a close at about the age when the animal reaches adulthood. Horseradish peroxidase was injected into two optic tecta (in a single animal) that contained units with permanently altered auditory spatial tuning. The positions of retrogradely labeled cells in the external nucleus of the inferior colliculus (ICX) were the same as those observed in control birds (Knudsen, E. I., and P. F. Knudsen (1983) J. Comp. Neurol. 218: 187-196). Thus, the changes in spatial tuning were not due to a shift in the topographic projection from the ICX to the optic tectum.(ABSTRACT TRUNCATED AT 400 WORDS)


Determination of stabilogram diffusion analysis coefficients and invariant density analysis parameters to understand postural stability associated with standing on anti-fatigue mats.
Prolonged standing has been associated with loss of balance, onset of low back pain symptoms and development of fatigue in lower extremity muscles in working populations. Although so far, it is unknown how individuals' postural stability is affected by standing on rigid versus cushioned platform but many industries are opting for anti-fatigue mats at workstations to reduce fall and injury related socio-economic cost. The goal of this study is to test SATECH's anti-fatigue mat for its effects on postural stability. A pilot test with seven healthy subjects (25–35 years old) has been conducted with a force plate to obtain kinetics of body when standing on two different platforms. The centers of pressure (COP) position of subjects were determined on rigid and anti-fatigue mats for quiet stance (each trial 60 seconds). In order to understand postural control along with dynamic or stochastic characteristics of the COP, stabilogram diffusion analysis (SDA) and Invariant density analysis methods are used. Subject specific patterns were seen in stabilogram diffusion plots and associated parameters in both conditions. We also found differences in some postural sway SDA parameters with anti-fatigue mats compared to rigid vinyl floor standing with open eyes condition. But no significant differences were found in sway IDA parameters. This work further provides insights whether anti fatigue mats can be helpful to workers involved with prolonged standing tasks.


Retinal complications after aqueous shunt surgical procedures for glaucoma.
To assess retinal complications and to identify risk factors for retinal complications following aqueous shunt procedures. Records of 38 consecutive aqueous shunt procedures that were performed on 36 patients at the Eye Institute of the Medical College of Wisconsin, Milwaukee, from June 1993 to March 1995 (minimum follow-up, 6 months) were reviewed. The mean +/- SD follow-up was 11.4 +/- 5.2 months (median, 10.5 months). Twelve patients (32%) had the following retinal complications: 4 serous choroidal effusions (10%) that required drainage, 3 suprachoroidal hemorrhages (8%), 2 vitreous hemorrhages (5%), 1 rhegmatogenous retinal detachment (3%), 1 endophthalmitis (3%), and 1 scleral buckling extrusion (3%). Surgical procedures for retinal complications were required in 8 (67%) of these 12 patients. Visual acuity decreased 2 lines or more in 9 (75%) of these 12 patients. The median onset of a postoperative retinal complication was 12.5 days, with 10 patients (83%) experiencing complications within 35 days. Serous choroidal effusions developed in 10 other patients (26%), and these effusions resolved spontaneously. Visual acuity decreased 2 lines or more in 2 (20%) of these additional 10 patients. Patients who experienced serious retinal complications were significantly older, had a higher rate of hypertension, and postoperative ocular hypotony. Serious retinal complications were distributed evenly among patients with Krupin valves with discs and Molteno and Baerveldt devices. Experience with the Ahmed glaucoma valve implant was limited. Aqueous shunt procedures may be associated with significant retinal complications and subsequent visual loss.


Does Resection of the Posterior Longitudinal Ligament Impact the Incidence of C5 Palsy After Cervical Corpectomy Procedures?: A Review of 459 Consecutive Cases.
Retrospective review. To evaluate key risk factors for the development of C5 palsy after cervical corpectomy, including resection of the posterior longitudinal ligament (PLL). Postoperative C5 palsy is a well-known complication after cervical spine surgery. It is unknown whether resection of the PLL affects the incidence of C5 palsy. We performed a retrospective review of 459 consecutive patients undergoing anterior cervical corpectomies over a 15-year period. Medical records were reviewed to gather demographic data, operative details, and the incidence of C5 palsy. We performed regression analyses to identify variables that predicted the development of C5 palsy. Our final analysis included 397 patients (females 51.4%, mean age 55.6 ± 11.6 yrs). Anterior corpectomy alone was performed in 255 (64.2%) patients, and combined anterior and posterior fusion was performed in 142 (35.8%) patients. Twenty-four patients (6.0%) developed C5 nerve palsy. Univariable regression demonstrated age greater than 65 (odds ratio, OR 2.7, 95% confidence interval, CI 1.2 to 6.3), corpectomy of three or more levels (OR 6.3, 95% CI 2.1 to 18.9), presence of ossification of the PLL (OR 4.3, 95% CI 1.6 to 11.7), and complete or partial resection of the PLL (OR 2.6, 95% CI 1.0 to 6.7) predicted development of C5 palsy. Multivariable regression demonstrated that the odds of getting C5 palsy with complete or partial resection of the PLL is 4.0 times (95% CI 1.5 to 10.5) higher compared with patients with an intact PLL. There were no significant differences in C5 palsy rates based on surgical approach (anterior vs. anterior plus posterior), sex, smoking status, or diabetes. Age greater than 65 years, corpectomy of three or more levels, presence of ossification of the PLL, and complete or partial resection of the PLL significantly predicted the development of C5 palsy. 4.


Transient paraparesis as a consequence of spontaneous spinal epidural haematoma.
A 78-year-old man presented to the accident and emergency department following sudden onset of back pain, with attendant loss of power and sensation in the lower half of his body and urinary retention. Onset occurred while he was seated in a chair and he fell as he tried to get up. Neurological examination revealed symmetrical lower motor weakness (MRC grade 4/5) in both lower limbs with downgoing plantar reflexes. There was a clear sensory boundary at D4 with loss of joint position and sensation to light touch below this level. Rectal examination was unremarkable. While a CT head scan was normal, spinal MRI showed multilevel disc desiccations in the dorsal and lumbar spine. Additionally, in the dorsal spine at D4/5, there was a thin plaque of epidural tissue representing an epidural haematoma. Within 48-72 h the patient gradually and spontaneously regained full power.


Accounting for intraclass correlations and controlling for baseline differences in a cluster-randomised evidence-based practice intervention study.
In health care and community-based intervention studies, cluster-randomised designs have been increasingly used because of administrative convenience, a desire to decrease treatment contamination, and the need to avoid ethical issues that might arise. While useful, cluster-randomised designs present challenges for data analysis. First, because of dependencies that exist among subjects within a cluster, methods that account for intra-class correlations have to be used. Second, on many occasions, because of unavailability of large numbers of clusters, lack of balance on baseline measures has to be carefully examined and appropriately controlled for. Two strategies are presented that can be used when analysing data from a cluster-randomised design; both account for baseline differences. Examples of these challenges are provided by a pain management intervention study designed to promote the adoption of evidence-based pain management practices. One approach involves use of a mixed model via SAS PROC MIXED. The other approach involves use of a marginal model: Generalised estimating equations using SAS PROC GENMOD. In cluster-randomised design, one must adjust for intra-class correlation when evaluating the intervention effect. Although the parameter estimates and their standard errors might be comparable with both random effect and marginal strategies for certain link functions (identity link or log link only), the interpretations are quite different and the two approaches are suitable for indicating answers to different questions. If differences are present concerning baseline measures between experimental and control groups, accounting for baseline measures is important. The choice between a mixed model or marginal approach should be dictated by whether the primary interest is a population or individual.


Reaction of Histone H1 with trans-Platinum Complexes and the Effect on DNA Platination.
Transplatin is an inactive platinum drug; however, a number of analogues, such as trans-EE and trans-PtTz, demonstrate promising antitumor activity in vitro and in vivo. Although the ultimate target is nuclear DNA, increasing evidence indicate that proteins also play important roles in the display of antitumor activity. The linker histone H1 is situated by the portal between the unwrapped DNA and the nucleosome core. Our recent study revealed that H1 can readily react with cisplatin, and the adducts tend to form ternary complexes with DNA. In this work, we have investigated the reaction of histone H1 with two antitumor-active trans-oriented complexes, trans-EE and trans-PtTz, and the effect of H1 upon the platination of DNA. The results show that trans-platinum drugs are much more reactive than cisplatin toward H1. Interestingly, in addition to the expected bidentate adducts (by displacement of the two labile chlorido ligands), also a tridentate adduct can be formed by displacement of one nonlabile carrier ligand of trans-EE or trans-PtTz. The trans-Pt/H1 adducts can then react with DNA and generate protein-Pt-DNA ternary complexes. Additionally, platinum can be transferred from trans-Pt/H1 adducts to DNA, generating binary trans-Pt/DNA complexes. Such a transfer of the platinum agent to DNA was not observed in the reaction of cisplatin. Furthermore, the detailed investigation carried out on a model peptide indicates that H1 promotes the DNA platination by trans- EE, while it reduces that of trans-PtTz and cisplatin. These results suggest that H1 can play a key role in the DNA platination and modulate the efficacy of different platinum agents.


'Otology at the crossroads' (the second Morell MacKenzie address).
It is an honour for an otological surgeon to be asked to deliver the second Sir Morell MacKenzie address. Thank you for the invitation and for that honour. Exactly 100 years ago MacKenzie, with Norris Wolfenden, founded 'The Journal of Laryngology and Rhinology'. 'Otology' was not added to the title until 1892, the year MacKenzie died. I shall return to this theme later. It is also exactly 100 years since, in 1887, MacKenzie was asked by Queen Victoria to examine her son-in-law, Frederick, Crown Prince of Germany. The controversy and subsequent recrimination which flowed from the management of Frederick's laryngeal lesion I shall leave to future lecturers to assess. It is as the founder of British laryngology that he should be remembered. As early as 1863 he had started the Metropolitan Free Dispensary for Diseases of the Throat and Loss of Voice, the first institution of its kind in the world. He wrote authoritatively on diseases of the throat and invented many instruments for the difficult art of indirect laryngeal and pharyngeal surgery. He received his Knighthood, also in 1887.


Role of artesunate in TGF‑β1‑induced renal tubular epithelial‑mesenchymal transdifferentiation in NRK‑52E cells.
The implications of epithelial‑mesenchymal transdifferentiation (EMT) have extended beyond the confines of renal fibrosis to renal tubulointerstitial fibrosis. It has been proposed that EMT may be one of the mechanisms involved in the pathogenesis of renal fibrosis. However, the underlying mechanisms remain unknown. Transforming growth factor (TGF)‑β1 is considered to be an important cytokine which regulates the transdifferentiation of tubular epithelial cells into myofibroblasts in renal tubulointerstitial fibrosis. In the present study, normal rat kidney tubular epithelial cells (NRK‑52E) were treated for 48 h with TGF‑β1 (5 ng/ml) and different concentrations of artesunate (ART; 0.01, 0.1 and 1 µg/ml). Western blotting, reverse transcription‑semi quantitative polymerase chain reaction analysis and immunofluorescence staining were used to evaluate the expression of bone morphogenetic protein (BMP)‑7, uterine sensitization‑associated gene (USAG)‑1, E‑cadherin, α‑smooth muscle actin (α‑SMA) and extracellular matrix collagen type I (Col I) mRNA. ART was able to attenuate renal injury in a unilateral ureteral obstruction model. However, its anti‑fibrotic effect remains to be elucidated. In the present study, it was observed that ART was able to ameliorate the TGF‑β1‑induced alterations in cellular morphology. In addition, ART inhibited the TGF‑β1‑induced USAG‑1 increase and the decrease in BMP‑7. Treatment with ART markedly attenuated the TGF‑β1‑induced upregulation of α‑SMA and downregulation of E‑cadherin. Additionally, ART was able to significantly attenuate the deposition of interstitial collagens, including Col I. The results of the present study further verified the therapeutic efficacy of ART in TGF‑β1‑induced renal interstitial fibrosis. These findings indicated that ART may hold the potential to prevent chronic kidney diseases via the suppression of USAG‑1 expression or by increasing BMP‑7 expression.


Predictors of successful ring pessary use in women with pelvic organ prolapse.
Pelvic organ prolapse (POP) is a common chronic health issue. Pessary rings are used for conservative management. To date, there is little evidence on objective anatomical findings as predictors of successful ring pessary management. To determine any association between history, clinical and four-dimensional translabial ultrasound (TLUS)/pelvic floor ultrasound examination and pessary success. From November 2013 to November 1015, all new patients presenting with symptomatic prolapse to a tertiary urogynaecological unit underwent an assessment including interview, clinical examination, that is, International Continence Society POPQ (pelvic organ prolapse quantification) and TLUS. Women with symptomatic prolapse were offered conservative management with a ring pessary. Those who agreed had a ring inserted that day. Successful trial of pessary use was defined as continued use for at least three months. Retrospective analysis of imaging data was performed blinded to other data. Statistical analysis was performed to assess the relationship between history, examination and imaging and pessary success. Of 525 patients seen during the inclusion period, 177 had symptomatic prolapse. One hundred and twenty-eight were offered a pessary, 89 accepted. Five had incomplete data, leaving 84. Forty- Two (50%) were still using the pessary at a three-month follow-up. Predictors associated with failure included being pre-menopausal (P = 0.031), a previous hysterectomy (P = 0.051), increasing genital hiatus and perineal body (Gh + Pb) (P = 0.013), posterior compartment prolapse (P = 0.027) and a larger hiatal area on Valsalva on TLUS (P = 0.049). Pre-menopausal status (P = 0.003), increasing Gh + Pb (P = 0.011) and previous hysterectomy (P = 0.001) remained significant on multivariate analysis. A history of previous hysterectomy is a predictor of pessary failure as are Gh+Pb on Valsalva and premenopausal status.


Training Programs on Endoscopic Scoring Systems for Inflammatory Bowel Disease Lead to a Significant Increase in Interobserver Agreement Among Community Gastroenterologists.
Endoscopic outcomes are increasingly used in clinical trials and in routine practice for inflammatory bowel disease [IBD] in order to reach more objective patient evaluations than possible using only clinical features. However, reproducibility of endoscopic scoring systems used to categorize endoscopic activity has been reported to be suboptimal. The aim of this study was to analyse the inter-rated agreement of non-dedicated gastroenterologists on IBD endoscopic scoring systems, and to explore the effects of a dedicated training programme on agreement. A total of 237 physicians attended training courses on IBD endoscopic scoring systems, and they independently scored a set of IBD endoscopic videos for ulcerative colitis [with Mayo endoscopic subscore], post-operative Crohn's disease [with Rutgeerts score] and luminal Crohn's disease (with the Simple Endoscopic Score for Crohn's Disease [SESCD] and Crohn's Endoscopic Index of Severity [CDEIS]). A second round of scoring was collected after discussion about determinants of discrepancy. Interobserver agreement was measured by means of the Fleiss' kappa [kappa] or intraclass correlation coefficient [ICC] as appropriate. The inter-rater agreement increased from kappa 0.51 (95% confidence interval [95% CI] 0.48-0.55) to 0.76 [95% CI 0.72-0.79] for the Mayo endoscopic subscore, and from 0.45 [95% CI 0.40-0.50] to 0.79 [0.74-0.83] for the Rutgeerts score before and after the training programme, respectively, and both differences were significant [P < 0.0001]. The ICC was 0.77 [95% CI 0.56-0.96] for SESCD and 0.76 [0.54- 0.96] for CDEIS, respectively, with only one measurement. The basal inter-rater agreement of inexperienced gastroenterologists focused on IBD management is moderate; however, a dedicated training programme can significantly impact on inter-rater agreement, increasing it to levels expected among expert central reviewers.


Simultaneous quantification of multiple urinary naphthalene metabolites by liquid chromatography tandem mass spectrometry.
Naphthalene is an environmental toxicant to which humans are exposed. Naphthalene causes dose-dependent cytotoxicity to murine airway epithelial cells but a link between exposure and human pulmonary disease has not been established. Naphthalene toxicity in rodents depends on P450 metabolism. Subsequent biotransformation results in urinary elimination of several conjugated metabolites. Glucuronide and sulfate conjugates of naphthols have been used as markers of naphthalene exposure but, as the current studies demonstrate, these assays provide a limited view of the range of metabolites generated from the parent hydrocarbon. Here, we present a liquid chromatography tandem mass spectrometry method for measurement of the glucuronide and sulfate conjugates of 1-naphthol as well as the mercapturic acids and N-acetyl glutathione conjugates from naphthalene epoxide. Standard curves were linear over 2 log orders. On column detection limits varied from 0.91 to 3.4 ng; limits of quantitation from 1.8 to 6.4 ng. The accuracy of measurement of spiked urine standards was -13.1 to + 5.2% of target and intra-day and inter-day variability averaged 7.2 (± 4.5) and 6.8 (± 5.0) %, respectively. Application of the method to urine collected from mice exposed to naphthalene at 15 ppm (4 hrs) showed that glutathione-derived metabolites accounted for 60-70% of the total measured metabolites and sulfate and glucuronide conjugates were eliminated in equal amounts. The method is robust and directly measures several major naphthalene metabolites including those derived from glutathione conjugation of naphthalene epoxide. The assays do not require enzymatic deconjugation, extraction or derivatization thus simplifying sample work up.


[The ecological, taxonomic and biotechnology aspects of research on bacteria and higher plants that produce biologically active substances].
The paper deals with the basic results of scientific activity of the Department of Antibiotics at the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine for the last two decades. Results of fundamental ecological and taxonomical investigations of bacteria of the Pseudomonas and Bacillus genera, study of regularities in formation of antibiotics and other biologically active substances by microorganism of these taxa and by higher plants have been presented; new antibiotics, probiotics and other biopreparations developed at the Department during this period have been characterized.


Improving Health for People Living With Heart Failure: Focus Group Study of Preconditions for Co-Production of Health and Care.
Co-production of health and care involving patients, families of patients, and professionals in care processes can create joint learning about how to meet patients' needs. Although barriers and facilitators to co-production have been examined previously in various health care contexts, the preconditions in Swedish chronic cardiac care contexts are yet to be explored. This study is set in the health system of the Swedish region of Jönköping County and is part of system-wide efforts to promote better health for persons with heart failure (HF). The objective of this study was to test the usefulness of the Capability, Opportunity, and Motivation Behavior (COM-B) model when assessing the barriers to and facilitators of co-production of health and care perceived by patients with HF, family members of patients with HF, and professionals in a Swedish chronic cardiac care context as a guide for subsequent initiatives. Data collection involved 1 focus group interview (FGI) with patients with HF (n=5), 1 FGI with family members of patients with HF (n=5), 1 FGI with professionals in primary care (n=7), and 1 FGI with professionals in cardiac care (n=4). In addition, patients with HF kept diaries of their thoughts regarding co-production. Using a deductive approach to content analysis, underpinned by the COM-B model, barriers and facilitators were categorized into capabilities, opportunities, and motivations to co-produce health and care. The participants showed limited understanding of co-production as a practice. They appeared to view it as a privilege to be offered to patients on top of traditional care and rarely as an approach for improving health care processes. The interviews revealed the limited health literacy among patients and the struggle of professionals to convey health information to these patients. Co-production was considered to be more resource-intensive than traditional care. Different expectations of stakeholders' roles were revealed: professionals expected older patients not to want to co-produce health and care, and all participants expected professionals to be in charge of health care services. The family members' position involved trying to balance their desire to support their relatives with understanding when, how, and with whom to co-produce. Presumed benefits motivated stakeholders: co-production was recognized to motivate patients to improve self-care. However, the participants recognized that motivation to get involved in health and care decisions varies over time among stakeholders. Co-production can be facilitated by the stakeholders' motivation. However, varying levels of understanding of co-production, patients' limited health literacy, unease with power sharing between patients and professionals, and resource constraints are barriers that need to be managed to promote co-produced care and better health for persons living with HF. Further research is warranted to explore how to co-produce health care services with patients with HF and how leaders can facilitate the inevitable cultural change it requires and represents.


Regulation of MyoD activity and muscle cell differentiation by MDM2, pRb, and Sp1.
Muscle cell differentiation is controlled by a complex set of interactions between tissue restricted transcription factors, ubiquitously expressed transcription factors, and cell cycle regulatory proteins. We previously found that amplification of MDM2 in rhabdomyosarcoma cells interferes with MyoD activity and consequently inhibits overt muscle cell differentiation (1). Recently, we found that MDM2 interacts with Sp1 and inhibits Sp1-dependent transcription and that pRb can restore Sp1 activity by displacing MDM2 from Sp1 (2). In this report, we show that forced expression of Sp1 can restore MyoD activity and restore overt muscle cell differentiation in cells with amplified MDM2. Furthermore, we show that pRb can also restore MyoD activity and muscle cell differentiation in cells with amplified MDM2. Surprisingly, we found that the MyoD-interacting domain of pRb is dispensable for this activity. We show that the C-terminal, MDM2-interacting domain of pRb is both necessary and sufficient to restore muscle cell differentiation in cells with amplified MDM2. We also show that the C-terminal MDM2-interacting domain of pRb can promote premature differentiation of proliferating myoblast cells. Our data support a model in which the pRb-MDM2 interaction modulates Sp1 activity during normal muscle cell differentiation.


The secreted protein ANGPTL2 promotes metastasis of osteosarcoma cells through integrin α5β1, p38 MAPK, and matrix metalloproteinases.
The tumor microenvironment can enhance the invasive capacity of tumor cells. We showed that expression of angiopoietin-like protein 2 (ANGPTL2) in osteosarcoma (OS) cell lines increased and the methylation of its promoter decreased with time when grown as xenografts in mice compared with culture. Compared with cells grown in normal culture conditions, the expression of genes encoding DNA demethylation-related enzymes increased in tumor cells implanted into mice or grown in hypoxic, serum-starved culture conditions. ANGPTL2 expression in OS cell lines correlated with increased tumor metastasis and decreased animal survival by promoting tumor cell intravasation mediated by the integrin α5β1, p38 mitogen-activated protein kinase, and matrix metalloproteinases. The tolloid-like 1 (TLL1) protease cleaved ANGPTL2 into fragments in vitro that did not enhance tumor progression when overexpressed in xenografts. Expression of TLL1 was weak in OS patient tumors, suggesting that ANGPTL2 may not be efficiently cleaved upon secretion from OS cells. These findings demonstrate that preventing ANGPTL2 signaling stimulated by the tumor microenvironment could inhibit tumor cell migration and metastasis.


Human bocavirus VP2 upregulates IFN-β pathway by inhibiting ring finger protein 125-mediated ubiquitination of retinoic acid-inducible gene-I.
Precise regulation of innate immunity is crucial for maintaining optimal immune responses against infections. Whereas positive regulation of IFN signaling elicits rapid type I IFNs, negative regulation is equally important in preventing the production of superfluous IFNs that can be hazardous to the host. The positive regulators of IFN pathway are known to be the main targets of viruses to antagonize the innate immune system. Whether viruses target the negative regulators of IFN pathway remains to be fully investigated. In this study, we report that the structural protein VP2 of human Bocavirus modulates IFN pathway by targeting the ring finger protein 125 (RNF125), a negative regulator of type I IFN signaling, which conjugates Lys(48)-linked ubiquitination to retinoic acid-inducible gene-I (RIG-I) and subsequently leads to the proteasome-dependent degradation of RIG-I. VP2 not only upregulated Sendai virus (SeV)-induced IFNB promoter activity, but also enhanced SeV-induced IFN-β production at both mRNA and protein levels. In agreement, the level of Ser(396)-phosphorylated IFN regulatory factor 3 stimulated by SeV was enhanced in the presence of VP2. Furthermore, VP2 was demonstrated to physically interact with RNF125, resulting in the reduction of RNF125-mediated ubiquitination and proteasome-dependent degradation of RIG-I. Additional study indicated that endogenous RIG-I degradation was decreased in VP2-expressing cells. Our study delineates a unique phenomenon for aberrant activation of IFN regulatory factor 3 pathway and may represent a new mechanism underlying viral manipulation of the host immune system.


Effects of alcohol taxes on alcohol-related disease mortality in New York State from 1969 to 2006.
The relationship of increased alcohol taxes to reductions in alcohol-related harm is well established. Few studies, however, have examined the effects of sudden decreases in alcohol tax rates or effects of narrow tax changes limited to specific beverage types. In the current study, we: (1) examine whether tax increases on spirits have similar effects in reducing alcohol-related disease mortality as increasing taxes on all types of alcoholic beverages simultaneously, and (2) evaluate effects of beer-specific tax decreases in New York State on mortality. We used a time-series, quasi-experimental research design, including non-alcohol deaths within New York State and other states' rates of alcohol-related disease mortality for comparison. The dataset included 456 monthly observations of mortality in New York State over a 38-year period (1969-2006). We used a random-effects approach and included several other important covariates. Alcohol-related disease mortality declined by 7.0% after a 1990 tax increase for spirits and beer. A spirits-only tax increase (in 1972) was not significantly associated with mortality but a data anomaly increased error in this effect estimate. Small tax decreases on beer between 1996 and 2006 had no measurable effect on mortality. Doubling the beer tax from $0.11 to $0.22 per gallon, a return to New York State's 1990 levels, would decrease deaths by an estimated 250 deaths per year. Excise tax increases on beer and spirits were associated with reductions in alcohol-related disease mortality. Modifying tax rates on a single beverage type does not appear to be as effective as doing so on multiple alcoholic beverages simultaneously. In New York, small decreases in beer taxes were not significantly associated with alcohol-related disease mortality.


Right ventricular afterload predicts long-term transition from parenteral to oral treprostinil in pulmonary arterial hypertension.
Despite the increasing trends, reports on long-term follow-up are limited on transitioning from parenteral to oral treprostinil therapy in patients with pulmonary arterial hypertension (PAH). We investigated both the effectiveness of parenteral to oral treprostinil transition and the characteristics associated with transition failure over a duration of two years. The study included 37 Group I functional class I and II patients with PAH on combination therapy. Patients were excluded if cardiac index ≤2.2 L/min/m2, right atrial pressure ≥11 mmHg, or 6-min walk distance ≤250 m. Patients were categorized as successful (STransition) or unsuccessful (UTransition) transition based on clinical stability, or a parenteral comparator (CParenteral) if they remained on parenteral therapy (no transition). All patients underwent two right heart catheterizations, one at enrollment and a second post transition. Of 24 total transition patients, 46% were classified as UTransition. UTransition occurred on average 577 days post transition. Both UTransition and STransition had similar hemodynamics at diagnosis and treprostinil dose before and after transition. Before transition, the pulmonary vascular resistance (PVR) was significantly higher in the UTransition (6.7 ± 2 WU) vs. STransition group (3.5 ± 1.5 WU). At follow-up catheterization, the UTransition group demonstrated further increases in PVR, greater than the CParenteral group, without recovery despite "rescue" therapy in the UTransition group. A pre-transition PVR of 4.16 WU discriminated the UTransition from the STransition group. While a subset of PAH patients on combination therapy may be safely transitioned from parenteral to oral treprostinil, caution should be exercised in patients with elevated baseline PVR to avoid irreversible destabilization.


The Sinus Membrane-Maxillary Lateral Wall Complex: Histologic Description and Clinical Implications for Maxillary Sinus Floor Elevation.
Maxillary sinus floor elevation has been documented as a safe and predictable procedure for gaining vertical bone height in the atrophic posterior maxillae. Conversely, there is a lack of basic research on the characteristics of the union between the sinus membrane (SM) and the bone. Clinical implications of an impaired union in healthy or pathologic membranes remain unknown. The objective of this study was to present a comprehensive histologic and morphologic description of the sinus membrane-lateral bone wall complex. In 14 fresh cadaver heads, 28 lateral wall sinus augmentation procedures were performed to obtain SM samples. Samples were assessed using hematoxylin-eosin, Masson trichrome, and toluidine blue staining and immunofluorescence and immunohistochemistry procedures. Specimens were coded and studied by a trained examiner using an optical microscope at ×4, ×10, ×40, and ×100 objectives. Thickness and inflammation status were assessed in these samples. Overall SM thickness of the samples was 0.40 ± 0.12 mm and was positively correlated to the inflammatory condition of the membranes. Such low values are the consequence of limited inflammation. Most of the fibers and cells in the deeper layers of the SM ran in a horizontal direction, oriented parallel to the underlying bone wall. In the immunohistochemistry study, 3 out of 7 samples showed a certain degree of nestin expression, suggesting osteogenic potential in spite of the elderly specimens. Large variations in thickness across the SM were found. These were noted to be partially correlated to the SM inflammatory status. The vast majority of the fibers were oriented parallel to the maxillary lateral wall, and only a few isolated areas showed a stronger perpendicular attachment. This might indicate the surpassing importance of the SM inflammatory status, operator skill, and other anatomical factors over the sinus membrane-maxillary lateral wall complex interface. Moreover, about half of the SM investigated were positive for nestin, indicating their osteogenic potential.


Serum retinoic acid, retinol and retinyl palmitate levels in patients with lung cancer.
Epidemiological studies have shown an inverse relationship between dietary vitamin A intake and the risk of developing lung cancer. The aim of this study was to investigate the vitamin A status in patients with lung cancer, by determining the serum levels of retinoic acid, retinol and retinyl palmitate. In total, 36 patients with lung cancer and 27 controls were assessed. Of the patients 14 had squamous cell carcinoma, 3 adenocarcinoma, 15 non-small cell lung cancer and 4 small cell lung cancer. Serum retinoic acid, retinol and retinyl palmitate levels were determined with HPLC and UV detection, after liquid extraction. Serum retinol levels did not differ between patients (733.5 +/- 326.4 ng/mL) and controls (734.5 +/- 337.1 ng/mL). The retinyl palmitate concentration tended to be lower in patients (14.3 +/- 9.7 ng/mL) than in controls (16.7 +/- 13.7 ng/mL). The serum retinoic acid levels were significantly lower in patients (1.9 +/- 0.6 ng/mL) than in controls (2.5 +/- 1.1 ng/mL, P < 0.05). A positive correlation was observed between the retinol and retinoic acid levels and retinyl palmitate and retinoic acid levels. The lower levels of retinoic acid in patients with lung cancer suggest there may be a deficiency or impairment in retinol metabolism in these patients. Further studies with larger numbers of patients are needed to evaluate the possible relationship between serum retinoid levels and lung cancer.


Impedance to gas transfer across the alveolar-capillary membrane in chronic cardiac failure.
One of the consequences of an elevation of the pulmonary capillary pressure in chronic heart failure is the occurrence of structural and functional changes at the level of the alveolar-capillary interface. These changes are called "stress failure" of the membrane, and consist of thickening of the interstitium, increase in capillary permeability to water and ions, and disruption of local regulatory mechanisms for gas exchange. Functional correlates are an augmented impedance to gas transfer (DL), a reduction in the alveolar-capillary membrane conduction (DM) and an increase in the volume of the pulmonary capillary blood (VC). DM and VC are the two subcomponents of DL. DM has been identified as the strongest respiratory predictor of oxygen uptake at peak exercise in patients with chronic heart failure, suggesting that impeded lung diffusion may significantly contribute to exercise limitation and ventilatory abnormalities. The evidence relating to the pathophysiological and clinical significance of the impairment in lung diffusion capacity in patients with chronic heart failure, as well as the response to treatment are the main subjects of this review.


Functional Outcomes Following Total Laryngectomy and Pharyngolaryngectomy: A 20-Year Single Center Study.
Laryngeal cancer accounts for 1% of all cancers in men and 0.3% of all cancers in women. Pharyngolaryngectomy (TPL) and total laryngectomy (TL) are central surgical techniques in the management of advanced laryngeal malignancies but are associated with significant morbidity. In addition, optimal reconstruction following TPL remains an area of active research. Here, we compared speech and swallowing outcomes following circumferential and partial pharyngeal resection alongside total laryngectomy in patients with laryngeal and hypolaryngeal tumors. We performed a systemic analysis of patient demographics, tumor characteristics, treatment modality, and pharyngeal reconstruction technique following TPL and TL, leveraging data collected over a 20-year period at a large tertiary referral center. Analyzing 155 patients the results show circumferential pharyngeal defects and prior radiotherapy have a significant impact on surgical complications. Pharyngeal resection carries a substantial risk of incurring impaired speech and swallowing in patients. Moreover, our results support poorer functional outcomes with more radical pharyngeal resections and show a clear trend toward worse swallowing outcomes in salvage surgery.


State-specific prevalence of cigarette smoking among adults, and children's and adolescents' exposure to environmental tobacco smoke -- United States, 1996.
In 1996, the prevalence of cigarette smoking was added to the list of nationally notifiable health conditions reported by states to CDC. The addition of a health-related behavior to the list of diseases and illnesses reflected the recognized role of tobacco use as the leading preventable cause of death in the United States. This report summarizes the 1996 prevalence of current smoking among adults in 49 states and the District of Columbia and presents state-specific estimates of environmental tobacco smoke (ETS) exposure for children and adolescents residing in homes where adults smoke. The findings indicate that state-specific smoking prevalence among adults varied twofold and that approximately 15 million children and adolescents were exposed to ETS in their home.


Human Coronaviruses: A Review of Virus-Host Interactions.
Human coronaviruses (HCoVs) are known respiratory pathogens associated with a range of respiratory outcomes. In the past 14 years, the onset of severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV) have thrust HCoVs into spotlight of the research community due to their high pathogenicity in humans. The study of HCoV-host interactions has contributed extensively to our understanding of HCoV pathogenesis. In this review, we discuss some of the recent findings of host cell factors that might be exploited by HCoVs to facilitate their own replication cycle. We also discuss various cellular processes, such as apoptosis, innate immunity, ER stress response, mitogen-activated protein kinase (MAPK) pathway and nuclear factor kappa B (NF-κB) pathway that may be modulated by HCoVs.


Criteria for early discharge after acute myocardial infarction: validation in a community hospital.
All patients admitted to a community hospital coronary care unit during an 18-month-period were studied to validate previously reported criteria for early hospital discharge after myocardial infarction. Factors present during the first four hospital days, which predict subsequent complications requiring urgent medical attention, were classified as either urgent or prognostic. Patients whose initial four days were marked by either no complications (81 patients) or prognostic complications (51 patients) are described. Only one patient in the group of 81 patients had a subsequent urgent complication. Four of 55 patients had late urgent complications. Persistent sinus tachycardia occurred during the first four days in all four of these 55 patients. Early hospital discharge would be feasible in the group with neither urgent nor prognostic complications. Further study of persistent sinus tachycardia is required to improve its predictive ability.


Cold-pressed and hot-pressed rapeseed oil: The effects of roasting and seed moisture on the antioxi- dant activity, canolol, and tocopherol level.
The paper looks at the levels of canolol, tocopherols and antioxidant activity in cold-pressed and hot-pressed rapeseed oils produced from seeds of various moisture levels (5%, 7.5%, and 10%). The paper also considers the effects of seed roasting on the levels of these compounds. The material used for the tests was rapeseed cv. Adrianna. The quality of the oils obtained is determined using peroxide and acid values. The levels of canolol and tocopherols are analyzed using HPLC. The DPPH radical-scavenging activity method for oil samples and phenolic extract from oils was used. It has been demonstrated that the oils produced from rapeseeds with a 5% moisture content, and in particular from cold-pressed oils, were characterized by the lowest peroxide values. Cold-pressed oils produced from rapeseeds with a 5% moisture content were characterized by higher levels of tocopherols and plastochromanol-8. In the case of hot-pressed oils, the highest levels of tocopherols were found in oils pro- duced from seeds with a 7.5% moisture content, and the greatest amount of PC-8 (more than 4 mg/100 g) was found in oils produced from seeds with a 10% moisture content. Hot-pressed oils have been shown to have higher levels of these compounds than cold-pressed oils. Both roasting and hot pressing led to an increase in the amount of canolol in the oils investigated. When analysing the antioxidant activity of the oils and phenolic extracts it was shown that phenolic compounds are responsible for approx. 10% of total antioxidant activity. Various levels of biologically active compounds were shown to be present in the rapeseed oil obtained from raw materials of a varying moisture content. The type of pressing process (cold-pressing or hot-pressing) and whether the seeds have undergone roasting has also been shown to affect the resulting oil and the level of native antioxidants it contains.


Purification and characterization of beta-glucosidase of Alcaligenes faecalis.
A cellobiose-utilizing bacterium isolated from sugar cane bagasse and identified as a strain of Alcaligenes faecalis (ATCC 21400) produced an inducible beta-glucoside-splitting enzyme. The enzyme was purified by a series of streptomycin and ammonium sulfate fractionations and by Sephadex and diethylaminoethyl column chromatography. The final preparation was purified 130-fold, with a recovery of about 10% of the initial enzyme activity. The enzyme had a wide pH range, with optimal activity at pH 6.0 to 7.0. The enzyme was stable in solution at pH 6.5 to 7.8 when kept at 30 C for 2 hr, but it was destroyed by temperatures above 55 C. At 58 and 60 C, the time required to inactivate 90% of the initial activity was 16 and 6.5 min, respectively. An activation energy of 9,500 cal/mole and a K(m) of 1.25 x 10(-4)m were obtained by using p-nitrophenyl beta-glucoside as a substrate. The K(i) value and hydrolysis of cellobiose by the enzyme indicated a high affinity of the enzyme for the cellobiose. The enzyme had its specificity on beta-glucosidic linkage and the rate of hydrolisis of glucosides depended upon the nature of the aglycon moiety. The inactivation studies showed the presence of sulfhydryl groups in the enzyme. The activity of the enzyme was easily destroyed by the Cu(++) and Hg(++) ions. The Michaelis-Menton relationship and the rate of heat inactivation indicated the presence of one type of noninteracting active site in the bacterial beta-glucosidase. Molecular weight of the enzyme was estimated by gel filtration (Sephadex G-200) and sucrose density gradient, and a value of 120,000 to 160,000 was obtained.


Evidence for serotonin synthesis, uptake, and release in dissociated rat sympathetic neurons in culture.
In the previous paper (Matsumoto et al., 1987), pharmacological evidence for secretion of serotonin (5-HT) by cultured sympathetic principal neurons was reported. Here, we present further evidence that 5-HT is a transmitter of some of these neurons under certain culture conditions, and can also be a "false" transmitter. Sympathetic principal neurons, dissociated from superior cervical ganglia (SCG) of newborn rats, were grown in cell culture. The presence of serotonin was demonstrated with high-performance liquid chromatography (HPLC), immunocytochemistry, and electrophysiological recording. Homogenates of cultures contained a substance that comigrated with authentic 5-HT in HPLC. A voltammogram for this substance was superimposed upon that for authentic 5-HT. When the cultures were examined with immunocytochemical staining, using the peroxidase-antiperoxidase technique, many neuronal processes contained 5-HT-like immunoreactive material. Many somata also contained 5-HT-like immunoreactivity when the neurons were grown in the presence of medium conditioned by heart cells (CM), but few somata stained above background in the absence of CM. Medium that contained a raised concentration of K+ (54 mM) or veratridine evoked Ca2+-dependent release of 5-HT, consistent with a neurotransmitter role for 5-HT in the cultures. In preliminary electrophysiological experiments on microcultures containing single sympathetic principal neurons and cardiac myocytes, a nonadrenergic excitatory (NAE) interaction was sometimes obtained after exposing the neurons to 100 microM 5-hydroxytryptophan. This interaction was sensitive to the serotonin blockers reserpine, methysergide, and gramine.(ABSTRACT TRUNCATED AT 250 WORDS)


Diarrhea Patterns and Climate: A Spatiotemporal Bayesian Hierarchical Analysis of Diarrheal Disease in Afghanistan.
Subject to a high burden of diarrheal diseases, Afghanistan is also susceptible to climate change. This study investigated the spatiotemporal distribution of diarrheal disease in the country and how associated it is with climate variables. Using monthly aggregated new cases of acute diarrhea reported between 2010 and 2016 and monthly averaged climate data at the district level, we fitted a hierarchical Bayesian spatiotemporal statistical model. We found aridity and mean daily temperature were positively associated with diarrhea incidence; every 1°C increase in mean daily temperature and 0.01-unit change in the aridity index were associated with a 0.70% (CI: 0.67%, 0.73%) increase and a 4.79% (CI: 4.30%, 5.26%) increase in the risk of diarrhea, respectively. Average annual temperature, on the other hand, was negatively associated, with a 3.7% (CI: 3.74%, 3.68) decrease in risk for every degree Celsius increase in annual average temperature. Temporally, most districts exhibited similar seasonal trends, with incidence peaking in summer, except for the eastern region where differences in climate patterns and population density may be associated with high rates of diarrhea throughout the year. The results from this study highlight the significant role of climate in shaping diarrheal patterns in Afghanistan, allowing policymakers to account for potential impacts of climate change in their public health assessments.


Expression of novel genes encoded by the paramyxovirus J virus.
Characterization of the J virus or, in keeping with recent nomenclature recommendations, J paramyxovirus (JPV) genome revealed a unique genome structure, consisting of eight genes in the order 3'-N-P/V/C-M-F-SH-TM-G-L-5'. The small hydrophobic (SH) protein and the transmembrane (TM) protein genes are predicted to encode proteins 69 and 258 aa in size, respectively. The 4401 nt attachment (G) protein gene, much larger than most other paramyxovirus attachment protein genes sequenced to date, encodes a putative 709 aa attachment protein and contains distally a second open reading frame (ORF-X) 2115 nt long. Experiments undertaken in this study were intended to confirm the sequence-based gene allocation of JPV and to determine if proteins encoded by the SH gene, the novel TM gene and ORF-X are expressed. Northern blot analyses carried out on mRNA purified from JPV-infected cells indicated that the putative transcription initiation and termination sequences flanking the SH and TM genes are functional, consistent with their allocation as discrete genes, although a high level of read-through was observed across almost all transcriptional boundaries. Probes specific to the G protein coding region and ORF-X both identified an mRNA species corresponding to the predicted length of the G gene, confirming sequence-based predictions. While the SH and TM proteins were both detected in infected cells, no evidence was found for the expression of ORF-X. Preliminary studies indicate that the novel TM protein is a type II glycosylated integral membrane protein, orientated with its C terminus exposed at the cell surface.


Monte Carlo-aided dosimetry of the theragenics TheraSeed model 200 103Pd interstitial brachytherapy seed.
A dosimetric study of a 103Pd seed for permanent interstitial brachytherapy, the Theragenics Corporation Model 200 (TheraSeed), has been undertaken utilizing Monte Carlo photon-transport (MCPT) simulations. All dosimetric quantities recommended by the American Association of Physicists in Medicine (AAPM) Task Group 43 (TG-43) [Med. Phys. 22, 209-234 (1995)] report have been calculated. This source contains graphite pellets coated with palladium metal, within which the radioactive 103Pd is distributed. Due to the significant influence of this metal coating thickness on the dose distribution, two coating thicknesses, 2.2 microm (light seed, representing currently available seeds) and 10.5 microm (heavy seed, representing reactor-produced seeds available before 1994), were analyzed. Quantities determined are the following: dose rate constant, radial dose function, anisotropy function, anisotropy factor, anisotropy constant, and "along and away" dose tables. The National Institute of Standards and Technology (NIST) Wide Angle Free Air Chamber (WAFAC) standard for air-kerma strength (SK,N99) was simulated, allowing a comparison to measured dosimetry data normalized to SKN99. The calculated dose-rate constants are 0.691 (light seed) and 0.694 cGy h(-1) U(-1) (heavy seed), where 1 U= 1 microGy x m2 x h(-1), in contrast to the recommended TG-43 value of 0.74 cGy h(-1) U(-1) and the value, 0.665 cGy h(-1) U(-1), recommended by AAPM report 69 [Med. Phys. 27, 634-642 (2000)]. Anisotropy constants (1/r2 weighted average, r > or = 1 cm) are 0.862 and 0.884 for the light seed and heavy seed, respectively. A generalization of the AAPM formalism [Med. Phys. 27, 634-642] for evaluating the time-dependent ratio of an administered-to-prescribed dose is presented. The findings of this study, in combination with 5% corrections applied to WAFAC measurements performed in 1999, imply that changes in the AAPM's recommended ratios as large as 6%, are indicated.


Corneal abnormalities early in the course of Fuchs' endothelial dystrophy.
Corneas with advanced Fuchs' endothelial dystrophy that require endothelial keratoplasty manifest anterior corneal structural and cellular abnormalities that have been associated with visual deficits before and after endothelial keratoplasty. In this study, we determined the onset of these abnormalities in the course of the disease. Cross-sectional study. Sixty-three eyes (39 subjects) with a range of severity of Fuchs' dystrophy and 25 eyes (13 subjects) with normal corneas. All corneas were examined using slit-lamp biomicroscopy, ultrasonic pachymetry, and confocal microscopy. The clinical grade of Fuchs' dystrophy was assessed according to the presence and extent of guttae and clinically evident edema and was categorized as mild (grades 1 and 2), moderate (grades 3 and 4), or advanced (grades 5 and 6). Normal corneas were devoid of any central guttae (grade 0). Corneal backscatter (haze) was measured from the confocal image light intensity profile. Stromal cell density and number and the presence of abnormal subepithelial cells were determined from confocal images. Comparisons between groups were made by using generalized estimating equation models. Anterior corneal backscatter, stromal cell density and number, presence of subepithelial cells, and central corneal thickness. Anterior corneal backscatter was 18% to 67% higher in eyes with moderate and advanced Fuchs' dystrophy compared with normal eyes (P ≤ 0.003); a similar trend was noted in mild Fuchs' dystrophy eyes compared with normal eyes (P = 0.08). Stromal cell density and the absolute number of stromal cells in the anterior 10% of the stroma were approximately 20% and 27% lower, respectively, in Fuchs' dystrophy (regardless of severity) compared with normal (P < 0.001). Abnormal subepithelial cells were visible in 9%, 19%, and 30% of corneas with mild, moderate, and advanced Fuchs' dystrophy, respectively. Only corneas with advanced Fuchs' dystrophy were thicker than normal (P < 0.001). Anterior corneal cellular and structural abnormalities begin early in the course of Fuchs' dystrophy, before the onset of clinically evident edema. The chronicity of these changes can explain their incomplete resolution after endothelial keratoplasty, and understanding the onset of these may help to determine the optimal time to intervene to achieve best outcomes.


EEG microstates are correlated with brain functional networks during slow-wave sleep.
Electroencephalography (EEG) microstates have been extensively studied in wakefulness and have been described as the "atoms of thought". Previous studies of EEG have found four microstates, i.e., microstates A, B, C and D, that are consistent among participants across the lifespan during the resting state. Studies using simultaneous EEG and functional magnetic resonance imaging (fMRI) have provided evidence for correlations between EEG microstates and fMRI networks during the resting state. Microstates have also been found during non-rapid eye movement (NREM) sleep. Slow-wave sleep (SWS) is considered the most restorative sleep stage and has been associated with the maintenance of sleep. However, the relationship between EEG microstates and brain functional networks during SWS has not yet been investigated. In this study, simultaneous EEG-fMRI data were collected during SWS to test the correspondence between EEG microstates and fMRI networks. EEG microstate-informed fMRI analysis revealed that three out of the four microstates showed significant correlations with fMRI data: 1) fMRI fluctuations in the insula and posterior temporal gyrus positively correlated with microstate B, 2) fMRI signals in the middle temporal gyrus and fusiform gyrus negatively correlated with microstate C, and 3) fMRI fluctuations in the occipital lobe negatively correlated with microstate D, while fMRI signals in the anterior cingulate and cingulate gyrus positively correlated with this microstate. Functional brain networks were then assessed using group independent component analysis based on the fMRI data. The group-level spatial correlation analysis showed that the fMRI auditory network overlapped the fMRI activation map of microstate B, the executive control network overlapped the fMRI deactivation of microstate C, and the visual and salience networks overlapped the fMRI deactivation and activation maps of microstate D. In addition, the subject-level spatial correlations between the general linear model (GLM) beta map of each microstate and the individual maps of each component yielded by dual regression also showed that EEG microstates were closely associated with brain functional networks measured using fMRI during SWS. Overall, the results showed that EEG microstates were closely related to brain functional networks during SWS, which suggested that EEG microstates provide an important electrophysiological basis underlying brain functional networks.


Dietary Iron Repletion Stimulates Hepatic Mobilization of Vitamin A in Previously Iron-Deficient Rats as Determined by Model-Based Compartmental Analysis.
Iron deficiency can result in hyporetinolemia and hepatic vitamin A (VA) sequestration. We used model-based compartmental analysis to determine the impact of iron repletion on VA metabolism and kinetics in iron-deficient rats. At weaning, Sprague-Dawley rats were assigned to either a VA-marginal diet (0.35 mg retinol equivalent/kg) with adequate iron (35 ppm, control group [CN]) or reduced iron (3 ppm, iron-deficient group [ID-]), with an equivalent average body weight for each group. After 5 wk, n = 4 rats from each group were euthanized for baseline measurements of VA and iron indices, and the remaining rats (n = 6 CN, n = 10 ID-) received an intravenous injection of 3H-labeled retinol in an emulsion as tracer to initiate the kinetic study. On day 21 after dosing, half of the ID- rats were switched to the CN diet to initiate iron repletion, referred to as the iron-repletion group (ID+). From the time of dosing, 34 serial blood samples were collected from each rat over a 92-d time course. Plasma tracer and tissue tracee data were fitted to 6- and 4-compartment models, respectively, to analyze the kinetic behavior of VA in all groups. Our mathematical model indicated that ID- rats exhibited a nearly 6-fold decrease in liver VA secretion and >4-fold reduction in whole-body VA utilization, compared with CN rats, whereas these perturbed kinetic behaviors were notably corrected in ID+ rats, close to those from the CN group. Iron repletion can remove the inhibitory effect that iron deficiency exerts on hepatic mobilization of VA and restore retinol kinetic parameters to values similar to that of never-deficient CN rats. Together with improvements in iron and VA indices, our results suggest that restoration of an iron-adequate diet is sufficient to improve VA kinetics after a previous state of iron deficiency.


Roles of endothelial and smooth muscle cell dysfunction and vasa vasorum in vasomotor disorders in ischemia with no obstructive coronary artery disease.
Recently, the importance has emerged of ischemia with no obstructive coronary artery disease (INOCA), for which endothelial and vascular smooth muscle cell (VSMC) dysfunctions and alterations in coronary vasa vasorum are involved. Regarding endothelial vasodilator functions, both endothelium-derived nitric oxide and endothelium-derived hyperpolarizing factor play important roles in modulating vascular tone, especially in the microcirculation. Recent studies have suggested systemic endothelial dysfunction in INOCA. Regarding VSMC dysfunction, Rho-kinase has been identified as a key molecular mechanism of VSMC hyperconstriction in INOCA. Finally, recent advances of coronary imaging have demonstrated the important role of altered adventitial vasa vasorum functions in INOCA.


Molecular cloning and characterization of an amphibian progesterone receptor from Rana dybowskii.
Progesterone plays a pivotal role in the regulation of reproduction in all vertebrates and binds to nuclear hormone receptor, one of ligand-dependent transcription factors. Although avian and mammalian progesterone receptors (PR) have been well characterized, detail structure and function of amphibian progesterone receptor in wild frog is poorly studied yet. Here we report the cloning and characterization of a novel progesterone receptor from the Korean frog, Rana dybowskii. The R. dybowskii progesterone receptor (dyPR, GenBank Accession No. AF431813) cDNA isolated from testis encodes a protein of 711 amino acids which shows approximately 60% overall identity with the Xenopus progesterone receptor. Reverse transcriptase-polymerase chain reaction (RT-PCR) demonstrates that dyPR is expressed in all the tissues examined. Electrophoretic mobility shift assays demonstrate that this receptor specifically binds to a progesterone response element (PRE), and transient transfection studies demonstrate that dyPR significantly activates the transcription of a PRE containing reporter element. Finally, confocal microscopy demonstrates the localization of this protein in nucleus, cytoplasm, and plasma membrane in transiently transfected CV-1 cell. These results indicate that dyPR cDNA encodes a classical progesterone receptor and molecular characterization of dyPR may provide us new information about the evolution of steroid hormone receptor.


Characterization of a vanillic acid non-oxidative decarboxylation gene cluster from Streptomyces sp. D7.
The genetics of non-oxidative decarboxylation of aromatic acids are poorly understood in both prokaryotes and eukaryotes. Although such reactions have been observed in numerous micro-organisms acting on a variety of substrates, the genes encoding enzymes responsible for these processes have not, to our knowledge, been reported in the literature. Here, the isolation of a streptomycete from soil (Streptomyces sp. D7) which efficiently converts 4-hydroxy-3-methoxybenzoic acid (vanillic acid) to 2-methoxyphenol (guaiacol) is described. Protein two-dimensional gel analysis revealed that several proteins were synthesized in response to vanillic acid. One of these was characterized by partial amino-terminal sequencing, leading to the cloning of a gene cluster from a genomic DNA lambda phage library, consisting of three ORFs, vdcB (602 bp), vdcC (1424 bp) and vdcD (239 bp). Protein sequence comparisons suggest that the product of vdcB (201 aa) is similar to phenylacrylate decarboxylase of yeast; the putative products of vdcC (475 aa) and vdcD (80 aa) are similar to hypothetical proteins of unknown function from various micro-organisms, and are found in a similar cluster in Bacillus subtilis. Northern blot analysis revealed the synthesis of a 2.5 kb mRNA transcript in vanillic-acid-induced cells, suggesting that the cluster is under the control of a single inducible promoter. Expression of the entire vdc gene cluster in Streptomyces lividans 1326 as a heterologous host resulted in that strain acquiring the ability to decarboxylate vanillic acid to guaiacol non-oxidatively. Both Streptomyces sp. strain D7 and recombinant S. lividans 1326 expressing the vdc gene cluster do not, however, decarboxylate structurally similar aromatic acids, suggesting that the system is specific for vanillic acid. This catabolic system may be useful as a component for pathway engineering research focused towards the production of valuable chemicals from forestry and agricultural by-products.


The projection diagram.
We describe a diagram, named the projection diagram (PD), that can be used for the interpretation of fringe projection operations in a similar way as the Holodiagram is used in holography and other branches in optics. It is obtained as a Moiré pattern between two spoke targets that mimic central projections, the same as those in a projection and observation system or two projection systems. N. Abramson [Academic, London (1981)] has already proposed its use in systems with two observation points (i.e., stereoscopic observation). By using this PD, several interesting features dealing with fringe projection are highlighted. Information on the sensitivity vector and the geometry of the contouring surfaces can be straightforwardly obtained in a graphical way. The effect of defocusing can also be included in the diagram.


Successful phased approach to a patient with synchronous traumatic descending aortic pseudoaneurysm and bronchial rupture.
A 34-year-old woman was referred to our center because of collapsed left lung and left main bronchial stenosis 1 week after a vehicular accident. Bronchoscopic observation revealed stenosis in the left main and lobar bronchus. Computed tomography (CT) angiography found traumatic descending aortic pseudoaneurysm after admission. Phased intervention strategy was adopted. The aortic rupture was repaired with endovascular stent firstly, followed by sleeve reconstruction of the left main bronchus through posterolateral thoracotomy 11 days later. She recovered uneventfully and resulted in an excellent long-term outcome. The diagnosis and treatment of this case is discussed.


Acute hyperammonemia activates branched-chain amino acid catabolism and decreases their extracellular concentrations: different sensitivity of red and white muscle.
Hyperammonemia is considered to be the main cause of decreased levels of the branched-chain amino acids (BCAA), valine, leucine, and isoleucine, in liver cirrhosis. In this study we investigated whether the decrease in BCAA is caused by the direct effect of ammonia on BCAA metabolism and the effect of ammonia on BCAA and protein metabolism in different types of skeletal muscle. M. soleus (SOL, slow-twitch, red muscle) and m. extensor digitorum longus (EDL, fast-twitch, white muscle) of white rat were isolated and incubated in a medium with or without 500 μM ammonia. We measured the exchange of amino acids between the muscle and the medium, amino acid concentrations in the muscle, release of branched-chain keto acids (BCKA), leucine oxidation, total and myofibrillar proteolysis, and protein synthesis. Hyperammonemia inhibited the BCAA release (81% in SOL and 60% in EDL vs. controls), increased the release of BCKA (133% in SOL and 161% in EDL vs. controls) and glutamine (138% in SOL and 145% in EDL vs. controls), and increased the leucine oxidation in EDL (174% of controls). Ammonia also induced a significant increase in glutamine concentration in skeletal muscle. The effect of ammonia on intracellular BCAA concentration, protein synthesis and on total and myofibrillar proteolysis was insignificant. The data indicates that hyperammonemia directly affects the BCAA metabolism in skeletal muscle which results in decreased levels of BCAA in the extracellular fluid. The effect is associated with activated synthesis of glutamine, increased BCAA oxidation, decreased release of BCAA, and enhanced release of BCKA. These metabolic changes are not directly associated with marked changes in protein turnover. The effect of ammonia is more pronounced in muscles with high content of white fibres.


Teaching home care to physicians during residency.
Residency is a pivotal time in a physician's education when lifetime patterns are often established. Thus, during residency physicians need to learn about home care and--more than just learn--become enthusiastic and committed to this type of health care. What can medical schools do to instill these values?