FTIR-ATR study of water uptake and diffusion through ion-selective membranes based on poly(acrylates) and silicone rubber.
For the first time, FTIR-ATR spectroscopy was used to study the water uptake and its diffusion in ion-selective membranes (ISMs) based on poly(acrylates) (PAs) and silicone rubber (SR), which are emerging materials for the fabrication of ISMs for ultratrace analysis. Three different types of PA membranes were studied, consisting of copolymers of methyl methacrylate with n-butyl acrylate, decyl methacrylate, or isodecyl acrylate. Numerical simulations with the finite difference method showed that in most cases the water uptake of the PA and SR membranes could be described with a model consisting of two diffusion coefficients. The diffusion coefficients of the PA membranes were approximately 1 order of magnitude lower than those of plasticized poly(vinyl chloride) (PVC)-based ISMs and only slightly influenced by the membrane matrix composition. However, the simulations indicated that during longer contact times, the water uptake of PA membranes was considerably higher than that for plasticized PVC membranes. Although the diffusion coefficients of the SR and plasticized PVC membranes were similar, the SR membranes had the lowest water uptake of all membranes. This can be beneficial in preventing the formation of detrimental water layers in all-solid-state ion-selective electrodes. With FTIR-ATR, one can monitor the accumulation of different forms of water, i.e., monomeric, dimeric, clustered, and bulk water.


Cancer Prevalence among Physicians in Korea: A Single Center Study.
There is little research regarding whether working as a physician affects cancer risk. Moreover, there is no research on cancer prevalence among physicians in Korea. This study utilized the Korea National Cancer Incidence Database to determine whether the prevalence of cancer among physicians differs from the prevalence of cancer within the general population. We analyzed the medical records of a representative sample of 382 doctors who underwent a health examination between 2010 and 2013 at a health examination center in a Ewha Womans University Medical Center.Cancer incidence was measured as cases that were eventually diagnosed as cancer according to a biopsy. We collected medical records from 382 physicians (mean age, 51.9±8.1 years) and calculated the standardized prevalence ratios compared to the general population. Thirty physicians (9 male and 21 female) were identified as having cancer. Physicians had a significantly higher prevalence of cancer compared to the general population.Cancer prevalence in male physicians was found to be 2.47 times higher than the prevalence expected within the general population (P=0.006). Among female physicians, cancer prevalence was 3.94 times higher than that in the general population (P<0.001). This study revealed that physicians had a higher prevalence of cancer compared to the general population in Korea, which suggests that there may be a problem present in the health care of physicians. Changes to the working environment of physicians will be needed to reduce the high prevalence of cancer among physicians.


Ultrastructure and morphology of antennal sensilla of the adult diving beetle Cybister japonicus Sharp.
The morphology and distribution of the antennal sensilla of adult diving beetle Cybister japonicus Sharp (Dytiscidae, Coleoptera), have been examined. Five types of sensilla on the antennae were identified by scanning electron microscope (SEM) and transmission electron microscope (TEM). Sensilla placodea and elongated s. placodea are the most abundant types of sensilla, distributing only on the flagellum. Both these types of sensilla carry multiple pore systems with a typical function as chemoreceptors. Three types of s. coeloconica (Type I-III) were also identified, with the characterization of the pit-in-pit style, and carrying pegs externally different from each other. Our data indicated that both type I and type II of s. coleconica contain two bipolar neurons, while the type III of s. coleconica contains three dendrites in the peg. Two sensory dendrites in the former two sensilla are tightly embedded inside the dendrite sheath, with no space left for sensilla lymph. There are no specific morphological differences in the antennal sensilla observed between males and females, except that the males have longer antennae and more sensilla than the females.


The effects of osmotic challenge on bronchial responsiveness to methacholine in non-asthmatic subjects.
We have measured the airway responses to methacholine on three days in sixteen non-asthmatic subjects. On the first day the methacholine challenge alone was performed. The other two days were randomized between pretreatment with distilled water or hypertonic saline administered by ultrasonic nebulizer. Distilled water and hypertonic saline did not affect baseline specific conductance (sGaw). Provocative dose producing a 35% fall in sGaw (PD35sGaw) was slightly reduced by both distilled water and hypertonic saline (15.83 to 8.55 mumol with distilled water and to 11.80 mumol with hypertonic saline). Six out of 16 subjects reached a plateau of maximal response with methacholine. The level of this plateau was not affected by pretreatment with distilled water or hypertonic saline. These results show that distilled water and 3.6% saline produced small increases in non-specific reactivity in normal subjects and confirm that substantial osmotic challenge does not change airway calibre in non-asthmatic subjects.


Noise covariance incorporated MEG-MUSIC algorithm: a method for multiple-dipole estimation tolerant of the influence of background brain activity.
This paper proposes a method of localizing multiple current dipoles from spatio-temporal biomagnetic data. The method is based on the multiple signal classification (MUSIC) algorithm and is tolerant of the influence of background brain activity. In this method, the noise covariance matrix is estimated using a portion of the data that contains noise, but does not contain any signal information. Then, a modified noise subspace projector is formed using the generalized eigenvectors of the noise and measured-data covariance matrices. The MUSIC localizer is calculated using this noise subspace projector and the noise covariance matrix. The results from a computer simulation have verified the effectiveness of the method. The method was then applied to source estimation for auditory-evoked fields elicited by syllable speech sounds. The results strongly suggest the method's effectiveness in removing the influence of background activity.


Ant species confer different partner benefits on two neotropical myrmecophytes.
The dynamics of mutualistic interactions involving more than a single pair of species depend on the relative costs and benefits of interaction among alternative partners. The neotropical myrmecophytes Cordia nodosa and Duroia hirsuta associate with several species of obligately symbiotic ants. I compared the ant partners of Cordia and Duroia with respect to two benefits known to be important in ant-myrmecophyte interactions: protection against herbivores provided by ants, and protection against encroaching vegetation provided by ants. Azteca spp., Myrmelachista schumanni, and Allomerus octoarticulatus demerarae ants all provide the leaves of Cordia and Duroia some protection against herbivores. However, Azteca and Allomerus provide more protection than does Myrmelachista to the leaves of their host plants. Although Allomerus protects the leaves of its hosts, plants occupied by Allomerus suffer more attacks by herbivores to their stems than do plants occupied by other ants. Relative to Azteca or Allomerus, Myrmelachista ants provide better protection against encroaching vegetation, increasing canopy openness over their host plants. These differences in benefits among the ant partners of Cordia and Duroia are reflected in the effect of each ant species on host plant size, growth rate, and reproduction. The results of this study show how mutualistic ant partners can differ with respect to both the magnitude and type of benefits they provide to the same species of myrmecophytic host.


The effects of Cinnamaldehyde on early brain injury and cerebral vasospasm following experimental subarachnoid hemorrhage in rabbits.
The neuroprotective and vasodilatory effects of cinnamaldehyde have been widely studied and documented. On the basis of these findings, we hypothesized that cinnamaldehyde exhibits therapeutic effects on subarachnoid hemorrhage-induced early brain injury and cerebral vasospasm. Thirty-two adult male New Zealand white rabbits were randomly divided into four groups of eight rabbits: control, subarachnoid hemorrhage, subarachnoid hemorrhage + vehicle, and subarachnoid hemorrhage + cinnamaldehyde. An intraperitoneal dose of 50 mg/kg cinnamaldehyde was administered 5 min following an intracisternal blood injection, followed by three further daily injections at identical doses. The animals were sacrificed 72 h after subarachnoid hemorrhage was induced. The cross-sectional areas and arterial wall thicknesses of the basilar artery were measured and hippocampal degeneration scores were evaluated. Treatment with cinnamaldehyde was effective in providing neuroprotection and attenuating cerebral vasospasm after subarachnoid hemorrhage in rabbits. It effectively increased the cross-sectional areas of the basilar artery and reduced the arterial wall thickness; in addition, hippocampal degeneration scores were lower in the cinnamaldehyde group. The findings of this study showed, for the first time to our knowledge, that cinnamaldehyde exhibits neuroprotective activity against subarachnoid hemorrhage-induced early brain injury and that it can prevent vasospasm. Potential mechanisms underlying the neuroprotection and vasodilation were discussed. Cinnamaldehyde could play a role in subarachnoid hemorrhage treatment.


The use of serum amylase and lipase in evaluating and managing blunt abdominal trauma.
In order to determine the usefulness of serum amylase and lipase in the initial evaluation and subsequent management of blunt abdominal trauma (BAT) patients, we collected serum amylase and lipase on 85 consecutive BAT patients at admission, hospital day 1, hospital day 3, and hospital day 7. Only one patient had a pancreatic injury. A total of 45 patients (53%) had at least one enzyme abnormality during the study. There was no correlation between amylase or lipase values and age, sex, type of injury, diagnostic tests, operation, and outcome. In a control group of nonabdominal-trauma patients with admit studies only, all enzyme values were normal. We conclude that serum amylase and lipase are randomly elevated in patients with nonpancreatic-BAT both initially and during subsequent hospitalization and are not useful clinical tools in these patients.


Unusual cellular disposition of the mitochondrial molecular chaperones Hsp60, Hsp70 and Hsp10.
A number of molecular chaperones in mammalian cells are localized in mitochondria and they are presumed to function mainly within this organelle. However, there is now compelling evidence that these chaperones are also localized at a variety of other sites/compartments in cells where they perform important functions. These proteins include: (i) the major chaperonin Hsp60 (or P1), which was identified in mammalian cells as a protein altered in mutants resistant to microtubule inhibitors and is involved in numerous functions at the cell surface and in other compartments; (ii) the Hspl0 or Cpn10 protein, which is a co-chaperone for Hsp60 in protein folding but also serves as an early pregnancy factor in maternal serum; and (iii) the mHsp70 protein, which plays a central role in mitochondrial protein import but is also important for cellular senescence (mortalin) and antigen presentation processes. The presence of these mitochondrial chaperones at specific extramitochondrial locations greatly broadens the range of functions that they can carry out in cells. However, these observations also raise important questions regarding the mechanisms by which these proteins reach these extramitochondrial locations. My paper will review some work in this area and discuss the significance of these results.


Training feed-forward neural networks with gain constraints
Inaccurate input-output gains (partial derivatives of outputs with respect to inputs) are common in neural network models when input variables are correlated or when data are incomplete or inaccurate. Accurate gains are essential for optimization, control, and other purposes. We develop and explore a method for training feedforward neural networks subject to inequality or equality-bound constraints on the gains of the learned mapping. Gain constraints are implemented as penalty terms added to the objective function, and training is done using gradient descent. Adaptive and robust procedures are devised for balancing the relative strengths of the various terms in the objective function, which is essential when the constraints are inconsistent with the data. The approach has the virtue that the model domain of validity can be extended via extrapolation training, which can dramatically improve generalization. The algorithm is demonstrated here on artificial and real-world problems with very good results and has been advantageously applied to dozens of models currently in commercial use.


A male with type I orofaciodigital syndrome.
We describe a three generation family with three females showing minor features of orofaciodigital syndrome type I and a severely affected male in the third generation. In addition to the classical features of OFD I, the male had bilateral duplication of the halluces, a feature diagnostic of OFD II, and an atrioventricular septal defect. Heart defects have not previously been reported in OFD I but have been reported in OFD II. It is important to examine the mothers of all male neonates with orofaciodigital syndrome with care before making a diagnosis of OFD II.


The MRN complex is transcriptionally regulated by MYCN during neural cell proliferation to control replication stress.
The MRE11/RAD50/NBS1 (MRN) complex is a major sensor of DNA double strand breaks, whose role in controlling faithful DNA replication and preventing replication stress is also emerging. Inactivation of the MRN complex invariably leads to developmental and/or degenerative neuronal defects, the pathogenesis of which still remains poorly understood. In particular, NBS1 gene mutations are associated with microcephaly and strongly impaired cerebellar development, both in humans and in the mouse model. These phenotypes strikingly overlap those induced by inactivation of MYCN, an essential promoter of the expansion of neuronal stem and progenitor cells, suggesting that MYCN and the MRN complex might be connected on a unique pathway essential for the safe expansion of neuronal cells. Here, we show that MYCN transcriptionally controls the expression of each component of the MRN complex. By genetic and pharmacological inhibition of the MRN complex in a MYCN overexpression model and in the more physiological context of the Hedgehog-dependent expansion of primary cerebellar granule progenitor cells, we also show that the MRN complex is required for MYCN-dependent proliferation. Indeed, its inhibition resulted in DNA damage, activation of a DNA damage response, and cell death in a MYCN- and replication-dependent manner. Our data indicate the MRN complex is essential to restrain MYCN-induced replication stress during neural cell proliferation and support the hypothesis that replication-born DNA damage is responsible for the neuronal defects associated with MRN dysfunctions.


Antibiotic Resistance and Usage-A Survey on the Knowledge, Attitude, Perceptions and Practices among the Medical Students of a Southern Indian Teaching Hospital.
Examining the knowledge, attitude, perceptions and practices (KAP) of the medical students regarding antibiotic resistance (ABR) and use can help us in devising suitable educational interventions for them, tailored according to their earlier held knowledge, beliefs, capabilities and experience. A cross sectional, questionnaire based survey was conducted among the second year medical students of a teaching hospital, whereby their KAP regarding antibiotic use and resistance was assessed by using a five point Likert scale, whose responses ranged from 'strongly agree' to 'strongly disagree,' 'always' to 'never and 'very important' to 'unimportant'. The data was analyzed by using simple descriptive statistics. Wherever it was relevant, the Chi-square test was used to determine any significant difference. The response rate was 100 per cent. The number of respondents who agreed that ABR was an important and a serious public health issue in our teaching hospital (n= 66, 68 per cent), was significantly less (p < 0.001) as compared to the number of respondents who agreed that ABR was an important and a serious issue which the country (n = 86, 88.65 per cent) and the world (n = 88, 90.7 per cent) were facing. Only 77.3 per cent (n= 75) of the respondents were aware that bacteria were not responsible for causing colds and flu, while the remaining 22.7 per cent (n = 22) were not knowledgeable about this fact. More than 80 per cent rated the adverse effect profile of the antibiotic and the risk of a superinfection as the important factors which deserved consideration. Cost of the antibiotic was considered to be an important factor deserving consideration by only 56.7 percent (n=55) of the participants. Our survey revealed that most of the students were aware of the antimicrobial resistance and its consequences. The only concern was their casual attitude regarding the antibiotic use. Further educational interventions are necessary to improve their understanding and perceptions on antibiotic resistance, as well as their attitude towards antibiotic use.


Multilaboratory study of the Biomic automated well-reading instrument versus MicroScan WalkAway for reading MicroScan antimicrobial susceptibility and identification panels.
This study compared the Biomic automated well reader results to the MicroScan WalkAway results for reading MicroScan antimicrobial susceptibility and identification panels at four different sites. Routine fresh clinical isolates and quality control (QC) organisms were tested at each study site. A total of 46,176 MicroScan panel drug-organism combinations were read. The Biomic category agreement for 3,117 Gram-negative bacteria was 98.4%, with 1.4% minor and 0.2% major discrepancies. The Biomic category agreement for 5,233 Gram-positive bacteria was 98.7%, with 0.9% minor, 0.3% major, and 0.1% very major errors. Essential agreement, defined as Biomic results that were within ±1 2-fold dilution of the MicroScan results, was 99.3% for Gram-negative bacteria and 98.3% for Gram-positive bacteria. Biomic reading of MicroScan identification panels provided an overall agreement (first- and second-choice organism match) of 99.5% with 846 Gram-negative isolates and 99.5% with 430 Gram-positive isolates. These results suggest that the Biomic automated reader can provide accurate reading of MicroScan panels and has the capability of a visual panel read for manual adjustment of results.


[Laser therapy for chronic pain in cancer patients].
A pilot controlled study included 266 cancer patients (stage IV) with chronic pain syndrome. Laser treatment significantly increased (18%) the effect of pain-relieving drug therapy. Due to application of this visual-analog laser procedure (WHO) carried out in three-stages, untoward side-effects fell 3-fold and were less marked. Life quality indices increased by 27.5-55.0%.


Suitable Data Representation for Abnormal Pattern Detection in Smart Home Environment.
Development in the area of sensor technologies and subsequently applications within the Internet of Things allows the implementation of systems for continuous monitoring of human's vital parameters and daily activities. This development is welcomed since the population aged 65 and over is constantly increasing. Moreover, the number of seniors living alone is also increasing. The monitoring systems can contribute to the safety and security of elderly people and allow them to stay at home where they are used to live as long as possible. Application of various sensor types raises questions on the most suitable sensor data representation, not losing useful information, and also on the design of detection and evaluation algorithms. In the paper, we briefly present several existing approaches and explain why we decided to use the basic ideas coming from the area of behaviour informatics.


Association between renal function and cardiovascular disease in patients with left ventricular hypertrophy. VIIDA study.
The association between renal dysfunction and cardiovascular risk in patients with hypertension and left ventricular hypertrophy (LVH) has not been specifically studied. The aim of this study was to analyze the association between renal function and the presence of cardiovascular disease in this group of patients. Hypertensive patients with electrocardiographic criteria for LVH were recruited in cardiology outpatient clinics from April 2003 until November 2004. Epidemiologic variables were determined, together with an estimation of the glomerular filtration rate by means of the Modification of Diet in Renal Disease (MDRD) and Crockoft-Gault equations. The population was classified according to the kidney disease stages of the National Kidney Foundation. A total of 3962 patients were included in the study, 47.6% of which were female, with a mean age of 67.2 years. The prevalence of established cardiovascular disease was higher in patients with a depressed glomerular filtration rate (68.3% vs 54.9%; P<.001). After adjusting for age, sex, body mass index, diabetes, smoking habits, and systolic and diastolic blood pressures, the stage of renal function was an independent predictor of the presence of cardiovascular disease (odds ratio, 1.5 [confidence interval, 1.19-2.02]; 2.1 [1.55-2.89]; and 2.6 [1.52-4.42], respectively, for stages 2, 3, 4-5, compared with stage 1). In hypertensive patients with electrocardiographic criteria for LVH, the determination of the glomerular filtration rate by the MDRD or Crockoft equations is easy and identifies a progressive and independent increase in cardiovascular risk.


A frameshift addition causes silencing of the delta-globin gene in an old world monkey, an anubis (Papio doguera).
A delta-globin gene of an anubis, Papio doguera, was cloned, and the nucleotide sequence of a region of about 2,000 bp including the gene was determined. In comparison with that of pSP3, a delta-globin clone from the gene library of a normal Japanese, the anubis delta-globin gene showed one base addition in the second exon at the codon for amino acid 55, generating a termination codon at the new 59th codon. The other differences found in the region analyzed were not considered to affect expression of the delta-globin gene. Therefore, it was concluded that this frameshift addition was at least one of the causes of silencing of the delta-globin gene in the anubis. In addition, from the sequence divergence in the delta-globin gene, the period of this silencing was estimated to be at most 4 million years, and the divergence of anubis occurred at most 30 million years ago.


Brazilian spotted fever in Espirito Santo, Brazil: description of a focus of infection in a new endemic region.
Although Brazilian spotted fever is known to occur in several adjacent states, infection with spotted fever group (SFG) rickettsiae has not previously been documented in the Brazilian state of Espirito Santo. We report a cluster of two proven and four suspected cases of Brazilian spotted fever in a small valley near Colatina, Espirito Santo. Four patients died without confirmatory serologic or pathologic studies. The two surviving patients had serologic and/or pathologic evidence of infection with SFG rickettsiae. Results of a survey done in 1991 confirmed the presence of antibodies reactive with SFG rickettsiae in the serum of dogs living in this region. Two of 25 dogs had elevated microimmunofluorescent antibody titers (1:64 and 1:256). Both seropositive dogs were pets of the index cases and lived in the valley where all six human cases were presumed to have acquired their infections. These cases and the results of the canine serosurvey suggest that a focus of virulent disease exists in an area not previously known to be endemic for SFG rickettsiae.


Spectral properties of topical retinoids prevent DNA damage and apoptosis after acute UV-B exposure in hairless mice.
We showed in a recent study that topical retinyl palmitate prevented UV-B-induced DNA damage and erythema in humans. Given that retinyl palmitate is a precursor of retinoic acid, the biological form of vitamin A that acts through nuclear receptors, we wondered whether these protective effects toward UV-B exposure were either receptor dependent or linked to other properties of the retinoid molecule such as its spectral properties. We determined the epidermal retinoid profile induced by topical retinoic acid in hairless mice and analyzed its effect on markers of DNA photodamage (thymine dimers) and apoptosis following acute UV-B exposure; we compared these effects to those induced by other natural topical retinoids (retinaldehyde, retinol and retinyl palmitate) which do not directly activate the retinoid receptors. We then analyzed the direct action of these retinoids on UV-B-induced DNA damage and apoptosis in cultured A431 keratinocytes. Topical retinoic acid significantly decreased (approximately 50%) the number of apoptotic cells, as well as the formation of thymine dimers in the epidermis of mice exposed to acute UV-B. Interestingly, the other topical retinoids decreased apoptosis and DNA damage in a similar way. On the other hand, neither retinoic acid nor the other retinoids interfered with the apoptotic process in A431 keratinocytes exposed to UV-B, whereas DNA photodamage was slightly decreased. We conclude that the decrease of apoptotic cells in hairless mouse epidermis following topical retinoids and UV-B irradiation reflects a protection of the primary targets of UV-B (DNA) by a mechanism independent of the activation of retinoid nuclear receptors, rather than a direct inhibition of apoptosis.


Molecular Hydrodynamics from Memory Kernels.
The memory kernel for a tagged particle in a fluid, computed from molecular dynamics simulations, decays algebraically as t^{-3/2}. We show how the hydrodynamic Basset-Boussinesq force naturally emerges from this long-time tail and generalize the concept of hydrodynamic added mass. This mass term is negative in the present case of a molecular solute, which is at odds with incompressible hydrodynamics predictions. Lastly, we discuss the various contributions to the friction, the associated time scales, and the crossover between the molecular and hydrodynamic regimes upon increasing the solute radius.


Ethnic variation in the clinical expression of idiopathic torsion dystonia.
Although childhood, limb-onset idiopathic torsion dystonia (ITD) is at increased prevalence among Jews, and Ashkenazi Jews with this form of ITD show linkage disequilibrium for a unique haplotype, little else is known about phenotypic or genetic variation in ITD among ethnic groups. We assessed clinical features in 786 ITD patients and determined the relative frequencies of various clinical forms of ITD in the major ethnic groups. Several differences in age-at-onset and site-at-onset distributions among groups were demonstrated. As expected, Ashkenazi Jews had an earlier age-at-onset distribution than did non-Jewish Caucasians. In the early-onset group, Jews were less likely to have cervical onset and more likely to have onset in the limbs than were non-Jewish Caucasians (p < 0.01). Among late-onset cases, Jews had less cervical onset and more cranial onset than did non-Jewish Caucasians (p < 0.01). African-Americans showed suggestive differences in both age at onset and site at onset as compared with the two caucasian groups, with an intermediate age at onset, a deficit of leg onset, and an excess of cranial and larynx onset. Subgroups of non-Jewish Caucasians showed no significant differences from each other. The observed ethnic variations in age at onset and site at onset of ITD suggest population differences in disease-causing alleles or loci. However, environmental influences on expression of ITD can not be excluded.


[Cholinergic neurons in the nuclear formations of the human fetal medulla oblongata].
The medulla oblongata of the human fetuses 22-27-week-old has been studied. Histochemical methods for cholinacetyltransferase (ChAT) and partly for acetylcholinesterase (AChE) have been used. Twenty-five nuclei have been described; four types of neurons with a low, middle, high and very high ChAT activity have been revealed in them. In the motor nuclei, neurons with a high, and in the sensitive nuclei--with a low ChAT activity predominate. In 13 nuclei a relative content of the cholinergic neurons has been calculated; it is low in the lateral reticular nucleus and in the Roller's nucleus (49-53%), high--in nuclei of the cranio-cerebral nerves (92-100%). In the motor neurons of the segmentary centers there is a certain correlation between the ChAT and AChE activity; in other neural centers (the inferior olive, the reticular substance nuclei) there is no such coincidence.


Purification and characterization of chromatin-bound DNA-dependent RNA polymerase I from parsley (Petroselinum crispum). Influence of nucleoside triphosphates.
The isolation and purification of DNA-dependent RNA polymerase I (EC 184.108.40.206) from parsley (Petroselinum crispum) callus cells grown in suspension culture is described. The enzyme was solubilized from isolated chromatin. Purification was achieved by using DEAE- and phospho-cellulose in batches, followed by column chromatography on DEAE- and phospho-cellulose (two columns) and density-gradient centrifugation. The highly purified enzyme was stable over several months. The properties of purified parsley RNA polymerase I were investigated. Optimum concentration for Mn2+ was 1 mM, and for Mg2+ 4-6 mM, Mn2+ was slightly more stimulatory than Mg2+. The enzyme was most active at low ionic strengths [10-20 mM-(NH4)SO4]. The influence of various phosphates was tested: pyrophosphate inhibited RNA polymerase at low concentrations, whereas orthophosphate had no effect on the enzyme activity. ADP was slightly inhibitory, and AMP had no effect on the enzyme reaction. Nucleoside triphosphates and bivalent cations in equimolar concentrations in the range 4-11 mM did not influence the RNA synthesis in vitro. Free nucleoside triphosphates in excess of this 1:1 ratio inhibited the enzyme activity, unlike free bivalent cations, which stimulated RNA polymerase I.


Fan-Beam Based Virtual Fluoroscopy for Navigated Catheterization in Interventional Radiology.
Personalized medicine implies reducing invasiveness of therapeutic procedures. Although interventional radiology proved a very interesting alternative to surgical procedures, it still raises concerns due to the irradiation dose received by the medical team (and by the patient). We propose a novel concept allowing to reduce very significantly the irradiation dose during the phases where tools inserted in the patient have to be tracked with respect to previously acquired images. This implies inserting a miniaturized X-ray detector in the tip of the tools, and reducing the dose by a "rotating collimator". We demonstrate that real-time processing of the signals allows accurate localization of the tip of the tools, with a dose reduction of at least ten times.


Choroid is thinner in inferior region of optic disks of normal eyes.
To determine the thickness of the choroid across the posterior pole of normal eyes. Twenty-eight clinic-based normal eyes (54.1 ± 20.0 years, -3.6 ± 4.1 diopter) were studied. The macula and retina around the optic disk were scanned with a spectral-domain optical coherence tomographic instrument. The retinal mapping program with the enhanced depth imaging technique was used, and the borders of the choroid were drawn manually in each optical coherence tomographic image. A choroidal thickness map was constructed by the built-in program, and the choroidal thickness in the different sectors of the Early Treatment Diabetic Retinopathy Study grid was measured and compared. The choroid inferior to the optic disk was significantly thinner than that in the other sectors of the outer ring of the Early Treatment Diabetic Retinopathy Study grid (superior, 196 ± 62 μm; inferior, 146 ± 47 μm; nasal, 183 ± 80 μm; and temporal, 193 ± 64 μm, P < 0.001). The mean choroidal thickness at the nasal sector of the macular region was significantly thinner than other regions of the Early Treatment Diabetic Retinopathy Study grid (superior, 268 ± 74 μm; inferior, 245 ± 73 μm; nasal, 190 ± 68 μm; temporal, 268 ± 63 μm; and central, 258 ± 88 μm; P < 0.05). Choroidal thickness maps showed that the thinner choroidal area spreads around the optic disk and the inferior part of the posterior pole. The thinner choroid inferior to the optic disk may be a natural anatomical architecture of normal eyes, and this area may be more susceptible to hypoxia or to elevated intraocular pressures.


Basement membrane and apocrine epithelial antigens in differential diagnosis between tubular carcinoma and sclerosing adenosis of the breast.
The distributions of defined basement membrane proteins in nine pure tubular carcinomas, 10 cases of sclerosing adenosis, and 15 ductal adenocarcinomas were compared. Sections of formalin fixed, paraffin embedded specimens were pretreated with pepsin and then immunostained for laminin, type IV collagen, and basement membrane proteoglycan, components specific for basement membranes. In sclerosing adenosis the tubules were surrounded by a continuous intact basement membrane composed of laminin, type IV collagen, and basement membrane proteoglycan, while the epithelium in the tubular carcinomas was negative for these proteins. The tumours were also analysed for the distribution of the apocrine epithelial antigen (AEA). In contrast to the benign lesions the tubular carcinomas expressed the AEA in a distinct non-polar fashion throughout the cell surface. In normal ducts and in adenosis the AEA was confined exclusively to the luminal surface. These studies suggest that there is a disturbance of cell polarity in tubular carcinomas. It is concluded that a combined analysis of basement membrane proteins and luminal surface antigens is a reliable and convenient way to differentiate between tubular carcinoma and sclerosing adenosis of the breast.


Membrane Targeting of Disheveled Can Bypass the Need for Arrow/LRP5.
The highly conserved Wnt signaling pathway regulates cell proliferation and differentiation in vertebrates and invertebrates. Upon binding of a Wnt ligand to a receptor of the Fz family, Disheveled (Dsh/Dvl) transduces the signal during canonical and non-canonical Wnt signaling. The specific details of how this process occurs have proven difficult to study, especially as Dsh appears to function as a switch between different branches of Wnt signaling. Here we focus on the membrane-proximal events that occur once Dsh is recruited to the membrane. We show that membrane-tethering of the Dsh protein is sufficient to induce canonical Wnt signaling activation even in the absence of the Wnt co-receptor Arrow/LRP5/6. We map the protein domains required for pathway activation in membrane tethered constructs finding that both the DEP and PDZ domains are dispensable for canonical signaling only in membrane-tethered Dsh, but not in untethered/normal Dsh. These data lead to a signal activation model, where Arrow is required to localize Dsh to the membrane during canonical Wnt signaling placing Dsh downstream of Arrow.


Brucellosis Suspicion is the Most Important Criterion for Diagnosis Particularly in Endemic Regions.
Brucellosis is a zoonotic infectious disease that remains endemic in developing countries. The purpose of this study is to emphasize the need for considering brucellosis as a diagnosis, since this disease has a high risk of complications among young patients when not treated appropriately. A total of 88 brucellosis cases with blood cultures that were positive for the pathogen were evaluated retrospectively in this study. The patients included 33 males (37.5%) and 55 females (62.5%) with a median age of 8.9 years (range: 5-14 years). A total of 43.1% (n=38) of the cases included occupational exposure to animals as a possible infection source. The consumption of raw milk products, especially cheese, was present in 52.2% (n=46) of the cases. Clinically, 55 of the cases were acute (62.5%), 23 of the cases were subacute (26.2%) and 10 of the cases were chronic (11.3%). The distribution of the joint pain complaints was as follows: 62.5% (n=55) of patients reported hip pain, 22.7% (n=20) of patients reported knee pain, 11.4% (n=10) of patients reported lumbar-back pain and 3.4% (n=3) of patients reported pain in other joints. A total of 59.1% (n=52) of the cases had been examined by another doctor at least once and mistreated. Complication rates and the rate of chronic infection increase with delayed diagnosis, and clinical doubt is the most important criterion for diagnosis, particularly in endemic regions.


Estrogen Receptor Expression in 21-Gene Recurrence Score Predicts Increased Late Recurrence for Estrogen-Positive/HER2-Negative Breast Cancer.
To identify the individual genes or gene modules that lead to the OncoptypeDx 21-gene recurrence score's reduced performance after 5 years and thereby identify indices of residual risk that may guide selection of patients for extended adjuvant therapy. We conducted a retrospective assessment of the relationship between (i) the individual genes and gene modules of the Recurrence Score and (ii) early (0-5 years) and late (5-10 years) recurrence rates in 1,125 postmenopausal patients with primary estrogen receptor-positive breast cancer treated with anastrozole or tamoxifen in the Arimidex, Tamoxifen, Alone or Combined (ATAC) randomized clinical trial. In the HER2-negative population (n = 1,009), estimates of recurrence risk were similar between years 0-5 and 5-10 for proliferation and invasion modules but markedly different for the estrogen module and genes within it (all split at the median): for low estrogen module, annual recurrence rates were similar across the two time windows (2.06% vs. 2.46%, respectively); for high estrogen module, annual rates were 1.14% versus 2.72%, respectively (P interaction = 0.004). Estrogen receptor transcript levels showed inverse prediction across the time windows: HR, 0.88 (0.73-1.07) and 1.19 (0.99-1.43), respectively (P interaction = 0.03). Similar time-, module-, and estrogen-dependent relationships were seen for distant recurrence. Patients with tumors with high estrogen receptor transcript levels benefit most from 5 years' endocrine therapy but show increased recurrence rates after 5 years and may benefit from extended therapy. Improved prognostic profiles may be created by considering period of treatment and follow-up time.


Ethnicity, deprivation and mortality due to 2009 pandemic influenza A(H1N1) in England during the 2009/2010 pandemic and the first post-pandemic season.
The relationship between risk of death following influenza A(H1N1)pdm09 infection and ethnicity and deprivation during the 2009/2010 pandemic period and the first post-pandemic season of 2010/2011 in England was examined. Poisson regression models were used to estimate the mortality risk, adjusted for age, gender, and place of residence. Those of non-White ethnicity experienced an increased mortality risk compared to White populations during the 2009/2010 pandemic [10·5/1000 vs. 6·0/1000 general population; adjusted risk ratio (RR) 1·84, 95% confidence interval (CI) 1·39-2·54] with the highest risk in those of Pakistani ethnicity. However, no significant difference between ethnicities was observed during the following 2010/2011 season. Persons living in areas with the highest level of deprivation had a significantly higher risk of death (RR 2·08, 95% CI 1·49-2·91) compared to the lowest level for both periods. These results highlight the importance of rapid identification of groups at higher risk of severe disease in the early stages of future pandemics to enable the implementation of optimal prevention and control measures for vulnerable populations.


[Involvement of miR-199a in cisplatin resistance of ovarian cancer cell through modulating expression of mTOR].
To study the effect of miR-199a on the sensitivity of OV2008 and C13* cells to cisplatin and investigate its mechanism of action. Real-time polymerase chain reaction(PCR) and Western blot were applied to detect the expression level changes of mammalian target of rapamycin (mTOR) in OV2008 and C13* cells before and after transfecting the cells with the mimics and inhibitor of miR-199a with Lipofectamine 2000.We constructed luciferase reporter vectors of mTOR and then transfected OV2008 cells with the mimics of miR-199a with Lipofectamine 2000 to study the regulation of miR-199a's downstream target genes. The expression level of mTOR was significantly higher in C13* than OV2008 cells (P<0.05). After OV2008 and C13* cells were transfected with the mimics and inhibitor of miR-199a, real-time PCR and Western blot showed that the mimics of miR-199a repressed the expression of mTOR; and the inhibitor of miR-199a promoted the expression of mTOR. After transfecting OV2008 cells with the mimics of miR-199a and luciferase reporter vectors of mTOR, renillaluciferase reporter gene analysis indicated the expression level of mTOR was negatively regulated by miR-199a. miR-199a regulates the sensitivity of ovarian cancer cells to cisplatin through modulating expression of mTOR, and involves in the cisplatin resistance process of ovarian cancer cells.


Concomitant tumor and minor histocompatibility antigen-specific immunity initiate rejection and maintain remission from established spontaneous solid tumors.
Nonmyeloablative hematopoietic cell transplantation can cure patients with hematologic malignancies but has reported limited success against solid tumors. This is possibly because of profound peripheral tolerance mechanisms and/or suboptimal tumor recognition by effector T lymphocytes. We report that in mice developing spontaneous prostate cancer, nonmyeloablative minor histocompatibility mismatched hematopoietic stem cell transplantation, and donor lymphocyte infusion of unmanipulated lymphocytes combined with posttransplant tumor-specific vaccination circumvents tumor-specific tolerance, allowing acute tumor rejection and the establishment of protective immunosurveillance. Although donor-derived tumor-specific T cells readily differentiated into effector cells and infiltrated the tumor soon after infusion, they were alone insufficient for tumor eradication, which instead required the concomitance of minor histocompatibiltiy antigen-specific CD8(+) T-cell responses. The establishment of protective immunosurveillance was best induced by posttransplant tumor-specific vaccination. Hence, these results provide the proof of principle that tumor-specific T-cell responses have to be harnessed together with minor histocompatibility responses and sustained by posttransplant tumor-specific vaccination to improve the efficacy of allotransplantion for the cure of solid tumors.


Functional activity of insulinoma cells (INS-1E) and pancreatic islets cultured in agarose cryogel sponges.
Here, we describe the preparation, structure, and properties of cryogel sponges, which represent a new type of macroporous biomaterial for tissue engineering. Cryogels were produced through freeze-thawing techniques, either from agarose alone or from agarose with grafted gelatin. The aim of this study was to evaluate agarose cryogel sponges as scaffolds for culturing both isolated pancreatic islets and insulinoma cells (INS-1E). In order to evaluate the effect of cell entrapment in artificial scaffolds, cell function reflected by insulin secretion and content was studied in cells cultivated for a 2-week period either in culture plastic plates or in cryogel sponge disks. Our results show that tumor-derived INS-1E cells grown either on plastic or on cryogels do not differ in their proliferation, morphology, insulin release, and intracellular insulin content. However, isolated pancreatic islets cultivated on cryogels sponge show 15-fold higher basal insulin secretion at 3.0 mM glucose than islets cultivated on plastic plates and fail to respond to stimulation with 16.7 mM glucose. In addition, these islets have about 2-fold lower insulin content compared to those grown in plastic plates. It is possible that the cell dysfunction noted in these in vitro experiments is due to the effect of the limited oxygen supply to the islets cultivated in cryogel sponge. Further in vivo studies are needed to clarify the nature of such an observation since according to previous reports, agarose and gelatin induce new vessel formation supporting enhanced oxygen supply.


The distribution of fitness effects in an uncertain world.
The distribution of fitness effects (DFE) among new mutations plays a critical role in adaptive evolution and the maintenance of genetic variation. Although fitness landscape models predict several key features of the DFE, most theory to date focuses on predictable environmental conditions, while ignoring stochastic environmental fluctuations that feature prominently in the ecology of many organisms. Here, we derive an extension of Fisher's geometric model that incorporates two common effects of environmental variation: (1) nonadaptive genotype-by-environment interactions (G × E), in which the phenotype of a given genotype varies across environmental contexts; and (2) random fluctuation of the fitness optimum, which generates fluctuating selection. We show that both factors cause a mismatch between the DFE within single generations and the distribution of geometric mean fitness effects (averaged over multiple generations) that governs long-term evolutionary change. Such mismatches permit strong evolutionary constraints-despite an abundance of beneficial fitness variation within single environmental contexts-and to conflicting DFE estimates from direct versus indirect inference methods. Finally, our results suggest an intriguing parallel between the genetics and ecology of evolutionary constraints, with environmental fluctuations and pleiotropy placing qualitatively similar limits on the availability of adaptive genetic variation.


Redistribution of neural phase coherence reflects establishment of feedforward map in speech motor adaptation.
Despite recent progress in our understanding of sensorimotor integration in speech learning, a comprehensive framework to investigate its neural basis is lacking at behaviorally relevant timescales. Structural and functional imaging studies in humans have helped us identify brain networks that support speech but fail to capture the precise spatiotemporal coordination within the networks that takes place during speech learning. Here we use neuronal oscillations to investigate interactions within speech motor networks in a paradigm of speech motor adaptation under altered feedback with continuous recording of EEG in which subjects adapted to the real-time auditory perturbation of a target vowel sound. As subjects adapted to the task, concurrent changes were observed in the theta-gamma phase coherence during speech planning at several distinct scalp regions that is consistent with the establishment of a feedforward map. In particular, there was an increase in coherence over the central region and a decrease over the fronto-temporal regions, revealing a redistribution of coherence over an interacting network of brain regions that could be a general feature of error-based motor learning in general. Our findings have implications for understanding the neural basis of speech motor learning and could elucidate how transient breakdown of neuronal communication within speech networks relates to speech disorders.


Photodynamic inactivation of cultured bladder tumor cells: a preliminary study of the effects of porphyrin aggregation.
This study was designed to determine the influence, if any, of intracellular porphyrin aggregation upon in vitro photosensitivity. HpD uptake conditions were manipulated in such a way as to yield three experimental groups, each containing equal levels of cellular porphyrins but having different porphyrin aggregation status. Aggregation status was determined through comparison of the spectral properties of intact cells. Subsequent exposure to red light resulted in a family of survival curves which generally corresponded to cellular HpD fluorescence yields and optical densities (indicators of aggregation status). These data suggest that porphyrin aggregation within the cell may be partially responsible for the observed differential photosensitivity following various uptake conditions.


Transplantation of iPSC-derived corneal endothelial substitutes in a monkey corneal edema model.
In order to provide regenerative therapy for millions of patients suffering from corneal blindness globally, we derived corneal endothelial cell substitute (CECSi) cells from induced pluripotent stem cells (iPSCs) to treat corneal edema due to endothelial dysfunction (bullous keratopathy). We developed an efficient xeno-free protocol to produce CECSi cells from both research grade (Ff-MH09s01 and Ff-I01s04) and clinical grade (QHJI01s04) iPSCs. CECSi cells formed a hexagonal confluent monolayer with Na, K-ATPase alpha 1 subunit expression (ATP1A1), tight junctions, N-cadherin adherence junction formation, and nuclear PITX2 expression, which are all characteristics of corneal endothelial cells. CECSi cells can be cryopreserved, and thawed CECSi cell suspensions also expressed N-cadherin and ATP1A1. Residual undifferentiated iPSCs in QHJI01s04-derived CECSi cells was below 0.01%. Frozen stocks of Ff-I01s04- and QHJI01s04-derived CECSi cells were transported, thawed and transplanted into a monkey corneal edema model. CECSi-transplanted eyes significantly reduced corneal edema compared to control group. Our results show a promising approach to provide bullous keratopathy patients with an iPS-cell-based cell therapy to recover useful vision.


Biological pest control using a model-based robust feedback.
Biological control is the artificial manipulation of natural enemies of a pest for its regulation to densities below a threshold for economic damage. The authors address the biological control of a class of pest population models using a model-based robust feedback approach. The proposed control framework is based on a recursive cascade control scheme exploiting the chained form of pest population models and the use of virtual inputs. The robust feedback is formulated considering the non-linear model uncertainties via a simple and intuitive control design. Numerical results on three pest biological control problems show that the proposed model-based robust feedback can regulate the pest population at the desired reference via the manipulation of a biological control action despite model uncertainties.


Validation of the Portuguese version of the Stanford presenteeism scale in nurses.
The SPS-6 is a tool that is widely used to test the characteristic of presenteeism, which is the ability to concentrate and work despite health problems. This concept refers to the phenomenon of people-despite complaining and being ill--going to work instead of resting and taking sick leave. The aim of this study was to assess the validity and reliability of the SPS-6. Internal consistency was examined by Cronbach's α. The Perceived Stress Scale was selected to determine the external validity of the SPS-6. Exploratory and confirmatory factor analyses were used to evaluate construct validity. The reliability analysis of all 126 registered nurses entered in the study showed satisfactory results (Cronbach's α = 0.83). External validity was evidenced by the significant negative correlation between the SPS-6 and the perceived stress scores. The factor analysis resulted in a two-component solution that conjointly accounted for 52.4% of the variance. A confirmatory factor analysis showed an adequate model fit. The Portuguese version of the SPS-6 displays the appropriate structure to measure the active employee's engagement at work.


Altered microRNA Expression Profiles and Regulation of INK4A/CDKN2A Tumor Suppressor Genes in Canine Breast Cancer Models.
microRNA (miRNA) expression profiling of cancer versus normal cells may reveal the characteristic regulatory features that can be correlated to altered gene expression in both human and animal models of cancers. In this study, the comprehensive expression profiles of the 277 highly characterized miRNAs from the canine genome were evaluated in spontaneous canine mammary tumor (CMT) models harboring defects in a group of cell cycle regulatory and potent tumor suppressor genes of INK4/CDKN2 family including p16/INK4A, p14ARF, and p15/INK4B. A large number of differentially expressed miRNAs were identified in three CMT cell lines to potentially target oncogenes, tumor suppressor genes and cancer biomarkers. A group of the altered miRNAs were identified by miRNA target prediction tools for regulation of the INK4/CDKN2 family tumor suppressor genes. miRNA-141 was experimentally validated for INK4A 3'-UTR target binding in the CMT cell lines providing an essential mechanism for the post-transcriptional regulation of the INK4A tumor suppressor gene in CMT models. A well-recognized group of miRNAs including miR-21, miR-155, miR-9, miR-34a, miR-143/145, and miR-31 were found to be altered in both CMTs and human breast cancer. These altered miRNAs might serve as potential targets for advancing the development of future therapeutic reagents. These findings further strengthen the validity and use of canine breast cancers as appropriate models for the study of human breast cancers.


[Heat shock proteins of freshwater protists and their involvement in adaptation to changes in the environmental salinity].
Changes in the level of heat shock proteins (HSP) in cells of freshwater protists, amoebae Amoeba proteus and ciliates Paramecium jenningsi, in response to changes in the environmental salinity were investigated. Changes in salinity levels were considered as a stress factor. The immunoblotting method revealed a polypeptide antigen cross-reacting with antibodies against bovine HSP70 in total protein extracts of both intact cells and cells subjected to salinity stress. The same polypeptide antigen was revealed in A. proteus cells subjected to heat shock. Therefore, it may be supposed that the polypeptide revealed after salinity shock is a heat shock protein related to the vertebrate HSP70. Under the impact of stress factor, well acclimated protists mostly spend their own previously accumulated HSP70. A conclusion is made that freshwater protists, living under conditions of increased salinity, appear to be preadapted to changes in environmental factors.


Distribution-free ROC analysis using binary regression techniques.
Receiver operating characteristic (ROC) regression methodology is used to identify factors that affect the accuracy of medical diagnostic tests. In this paper, we consider a ROC model for which the ROC curve is a parametric function of covariates but distributions of the diagnostic test results are not specified. Covariates can be either common to all subjects or specific to those with disease. We propose a new estimation procedure based on binary indicators defined by the test result for a diseased subject exceeding various specified quantiles of the distribution of test results from non-diseased subjects with the same covariate values. This procedure is conceptually and computationally simplified relative to existing procedures. Simulation study results indicate that the approach has fairly high statistical efficiency. The new ROC regression methodology is used to evaluate childhood measurements of body mass index as a predictive marker of adult obesity.


Structural determinant of human La protein critical for internal initiation of translation of hepatitis C virus RNA.
Human La protein has been implicated in facilitating internal ribosome entry site (IRES)-mediated translation of hepatitis C virus (HCV). Earlier, we demonstrated that the RNA recognition motif (RRM) encompassing residues 112 to 184 of La protein [La (112-184)] interacts with the HCV IRES near the initiator AUG codon. A synthetic peptide, LaR2C (24-mer), derived from La RRM (112-184), retains RNA binding ability, competes with La protein binding to the HCV IRES, and inhibits translation. The peptide interferes with the assembly of 48S complexes, resulting in the accumulation of preinitiation complexes that are incompetent for the 60S ribosomal subunit joining. Here, nuclear magnetic resonance spectroscopy of the HCV IRES-bound peptide complex revealed putative contact points, mutations that showed reduced RNA binding and translation inhibitory activity. The residues responsible for RNA recognition were found to form a turn in the RRM (112-184) structure. A 7-mer peptide comprising this turn showed significant translation inhibitory activity. The bound structure of the peptide inferred from transferred nuclear Overhauser effect experiments suggests that it is a beta turn. This conformation is significantly different from that observed in the free RRM (112-184) NMR structure, suggesting paths toward a better-stabilized mimetic peptide. Interestingly, addition of hexa-arginine tag enabled the peptide to enter Huh7 cells and showed inhibition of HCV IRES function. More importantly, the peptide significantly inhibited replication of the HCV monocistronic replicon. Elucidation of the structural determinant of the peptide provides a basis for developing small peptidomimetic structures as potent anti-HCV therapeutics.


An anticancer Os(II) bathophenanthroline complex as a human breast cancer stem cell-selective, mammosphere potent agent that kills cells by necroptosis.
Conventional chemotherapy is mostly effective in the treatment of rapidly-dividing differentiated tumor cells but has limited application toward eliminating cancer stem cell (CSC) population. The presence of a very small number of CSCs may contribute to the development of therapeutic resistance, metastases, and relapse. Thus, treatment failure by developing novel anticancer drugs capable of effective targeting of CSCs is at present a major challenge for research focused on chemotherapy of cancer. Here, we show that Os(II) complex 2 [Os(η6-pcym)(bphen)(dca)]PF6 (pcym = p-cymene, bphen = bathophenanthroline, and dca = dichloroacetate), is capable of efficient and selective killing CSCs in heterogeneous populations of human breast cancer cells MCF-7 and SKBR-3. Notably, its remarkable submicromolar potency to kill CSCs is considerably higher than that of its Ru analog, [Ru(η6-pcym)(bphen)(dca)]PF6 (complex 1) and salinomycin, one of the most selective CSC-targeting compounds hitherto identified. Furthermore, Os(II) complex 2 reduces the formation, size, and viability of three-dimensional mammospheres which more closely reflect the tumor microenvironment than cells in traditional two-dimensional cultures. The antiproliferation studies and propidium iodide staining using flow cytometry suggest that Os(II) complex 2 induces human breast cancer stem cell death predominantly by necroptosis, a programmed form of necrosis. The results of this study demonstrate the promise of Os(II) complex 2 in treating human breast tumors. They also represent the foundation for further preclinical and clinical studies and applications of Os(II) complex 2 to comply with the emergent need for human breast CSCs-specific chemotherapeutics capable to treat chemotherapy-resistant and relapsed human breast tumors.


Detecting Racial/Ethnic Health Disparities Using Deep Learning From Frontal Chest Radiography.
The aim of this study was to assess racial/ethnic and socioeconomic disparities in the difference between atherosclerotic vascular disease prevalence measured by a multitask convolutional neural network (CNN) deep learning model using frontal chest radiographs (CXRs) and the prevalence reflected by administrative hierarchical condition category codes in two cohorts of patients with coronavirus disease 2019 (COVID-19). A CNN model, previously published, was trained to predict atherosclerotic disease from ambulatory frontal CXRs. The model was then validated on two cohorts of patients with COVID-19: 814 ambulatory patients from a suburban location (presenting from March 14, 2020, to October 24, 2020, the internal ambulatory cohort) and 485 hospitalized patients from an inner-city location (hospitalized from March 14, 2020, to August 12, 2020, the external hospitalized cohort). The CNN model predictions were validated against electronic health record administrative codes in both cohorts and assessed using the area under the receiver operating characteristic curve (AUC). The CXRs from the ambulatory cohort were also reviewed by two board-certified radiologists and compared with the CNN-predicted values for the same cohort to produce a receiver operating characteristic curve and the AUC. The atherosclerosis diagnosis discrepancy, Δvasc, referring to the difference between the predicted value and presence or absence of the vascular disease HCC categorical code, was calculated. Linear regression was performed to determine the association of Δvasc with the covariates of age, sex, race/ethnicity, language preference, and social deprivation index. Logistic regression was used to look for an association between the presence of any hierarchical condition category codes with Δvasc and other covariates. The CNN prediction for vascular disease from frontal CXRs in the ambulatory cohort had an AUC of 0.85 (95% confidence interval, 0.82-0.89) and in the hospitalized cohort had an AUC of 0.69 (95% confidence interval, 0.64-0.75) against the electronic health record data. In the ambulatory cohort, the consensus radiologists' reading had an AUC of 0.89 (95% confidence interval, 0.86-0.92) relative to the CNN. Multivariate linear regression of Δvasc in the ambulatory cohort demonstrated significant negative associations with non-English-language preference (β = -0.083, P < .05) and Black or Hispanic race/ethnicity (β = -0.048, P < .05) and positive associations with age (β = 0.005, P < .001) and sex (β = 0.044, P < .05). For the hospitalized cohort, age was also significant (β = 0.003, P < .01), as was social deprivation index (β = 0.002, P < .05). The Δvasc variable (odds ratio [OR], 0.34), Black or Hispanic race/ethnicity (OR, 1.58), non-English-language preference (OR, 1.74), and site (OR, 0.22) were independent predictors of having one or more hierarchical condition category codes (P < .01 for all) in the combined patient cohort. A CNN model was predictive of aortic atherosclerosis in two cohorts (one ambulatory and one hospitalized) with COVID-19. The discrepancy between the CNN model and the administrative code, Δvasc, was associated with language preference in the ambulatory cohort; in the hospitalized cohort, this discrepancy was associated with social deprivation index. The absence of administrative code(s) was associated with Δvasc in the combined cohorts, suggesting that Δvasc is an independent predictor of health disparities. This may suggest that biomarkers extracted from routine imaging studies and compared with electronic health record data could play a role in enhancing value-based health care for traditionally underserved or disadvantaged patients for whom barriers to care exist.


LC-MS/MS isomeric profiling of permethylated N-glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients.
Early stage detection and cancer treatment demand the identification of reliable biomarkers. Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. Herein, an LC-MS/MS-based analysis was conducted to define the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients. The haptoglobin samples were extracted from serum using an antibody-immobilized column prior to the release of N-glycans. A comparison of non-isomeric and isomeric permethylated glycan forms was achieved using C18 and porous graphitic carbon (PGC) columns, respectively. In the case of C18-LC-MS/MS analysis, 25 glycan structures were identified of which 10 sialylated structures were found to be statistically significant between the two cohorts. Also, 8 out of 34 glycan structures identified by PGC-LC-MS/MS were found to be statistically significant, suggesting that isomeric distributions of a particular glycan composition were different in abundances between the two cohorts. The glycan isoform patterns distinguished early stage HCC from cirrhotic patients. Both retention times and tandem mass spectra were utilized to determine the specific isomeric glycan structures. All of the glycan isomers, which were statistically significant, were either branch fucosylated or composed of α-2,6 linked sialic acid moieties. The result of this study demonstrates the potential importance of isomeric separation for defining disease prompted aberrant glycan changes. The levels of several glycan isoforms effectively distinguished early stage HCC from cirrhosis.


Effects of call on sleep and mood in internal medicine residents.
Residents on call experience decreased total sleep time (TST) and increased dysphoria. This study monitored changes in mood and sleepiness for 3 post-call days. Fifty-two internal medicine residents participated in the study. The residents wore actigraphs for the 4 to 9 days of the study. Each morning resident completed mood scales, a sleepiness scale, and estimated their prior night TST. The residents were on a 1-in-4 schedule. Call decreased subjective- and actigraphy-derived TST to less than 4 hr. During the 3 days post call, mood measures improved. Tension, depression, and anxiety stabilized on the first post-call day following the first night of off-call sleep during which the residents obtained about 7 hr of sleep. Vigor, fatigue, and confusion stabilized on the second post-call day. The Epworth Sleepiness Scale dropped to less than 11 after 1 post-call night and continued to decrease up to 3 post-call days. The effects of call linger past the first recovery night. For these residents, recovery sleep appeared inadequate, and the negative effects of call persisted across succeeding off-call days. Thus, for these residents on a 1-in-4 schedule, call affects their mood for much of the time when off call and potentially their personal and professional interactions during this period as well.


Differences in Granule Morphology yet Equally Impaired Exocytosis among Cytotoxic T Cells and NK Cells from Chediak-Higashi Syndrome Patients.
Chediak-Higashi syndrome (CHS) is caused by autosomal recessive mutations in LYST, resulting in enlarged lysosomal compartments in multiple cell types. CHS patients display oculocutaneous albinism and may develop life-threatening hemophagocytic lymphohistiocytosis (HLH). While NK cell-mediated cytotoxicity has been reported to be uniformly defective, variable defects in T cell-mediated cytotoxicity has been observed. The latter has been linked to the degree of HLH susceptibility. Since the discrepancies in NK cell- and T cell-mediated cellular cytotoxicity might result from differences in regulation of cytotoxic granule release, we here evaluated perforin-containing secretory lysosome size and number in freshly isolated lymphocytes from CHS patients and furthermore compared their exocytic capacities. Whereas NK cells from CHS patients generally contained a single, gigantic perforin-containing granule, cytotoxic T cells predominantly contained several smaller granules. Nonetheless, in a cohort of 21 CHS patients, cytotoxic T cell and NK cell granule exocytosis were similarly impaired upon activating receptor stimulation. Mechanistically, polarization of cytotoxic granules was defective in cytotoxic lymphocytes from CHS patients, with EEA1, a marker of early endosomes, mislocalizing to lysosomal structures. The results leads to the conclusion that lysosome enlargement corresponds to loss of distinct organelle identity in the endocytic pathway, which on a subcellular level more adversely affects NK cells than T cells. Hence, vesicular size or numbers do not per se dictate the impairment of lysosomal exocytosis in the two cell types studied.


Effectiveness of quality improvement: learning from evaluations.
The effectiveness of many quality improvement interventions has been studied, and research suggests that most have highly variable effects which depend heavily on the context in which they are used and the way they are implemented. This has three important implications. Firstly, it means that the approach to quality improvement used in an organisation probably matters less than how and by whom it is used. Rather than taking up, trying, and then discarding a succession of different quality improvement techniques, organisations should probably choose one carefully and then persevere to make it work. Secondly, future research into quality improvement interventions should be directed more at understanding how and why they work--the determinants of effectiveness--rather than measuring whether they work. Thirdly, some element of evaluation should be incorporated into every quality improvement programme so that its effectiveness can be monitored and the information can be used to improve the systems for improvement.


Single-dose therapy with streptomycin and sulfametopyrazine for bacteriuria during pregnancy.
One hundred and sixty-three patients with bacteriuria in pregnancy were treated with a single dose of antibiotic. Four regimens were used: sulfadoxine (sulphormethoxine) 2 g., sulfametopyrazine 2 g., streptomycin 1 g., and a combination of sulfametopyrazine 2 g. and streptomycin 1 g. The highest cure rate of 77% was achieved with streptomycin and sulfametopyrazine. The long-acting sulphonamides alone gave a cure rate of 55% while streptomycin alone eradicated 43% of the infections. Despite the high cure rate with combined therapy bacterial resistance developed in some cases.


Anesthesia for endolaryngeal surgery.
A method of anesthesia for endolaryngeal surgery that dispenses with the use of an indwelling endotracheal tube with consequent improvement in the surgical conditions is described. This method further permits the manipulation of the surgeon's laryngoscope and the use of endolaryngeal surgery to improve the compromised or potentially compromised laryngeal airway without serious risk and with avoidance in some circumstances of preliminary tracheostomy.


Orthoreovirus infection and concurrent cryptosporidiosis in rough green snakes (Opheodrys aestivus): pathology and identification of a novel orthoreovirus strain via polymerase chain reaction and sequencing.
Reoviruses are nonenveloped, segmented, double-stranded RNA viruses capable of infecting a wide range of invertebrate, vertebrate, fungus, and plant hosts. Though sporadic infection has been reported in a variety of reptilian species, infection of rough green snakes (Opheodrys aestivus) has not been previously described. Five wild-caught, adult rough green snakes were obtained by a zoological institution. Clinical deterioration was first noted in all snakes after 3 weeks in quarantine. Despite treatment, clinical decline progressed, and all 5 snakes died or were euthanized by 48 days post-arrival. Moderate, multifocal, acute, necrotizing hepatitis with hepatocellular syncytia was diagnosed in 1 snake. Two additional snakes had severe, diffuse, subacute to chronic pancreatitis. All 5 snakes had gastroenteric cryptosporidiosis. Electron microscopic examination of liver from the snake with hepatic lesions revealed scattered hepatocytes containing 1 or more intranuclear clusters of approximately 90 nm in diameter viral particles arranged in loose arrays. Polymerase chain reaction (PCR) amplification of a segment of the reovirus RNA-dependent RNA polymerase gene was performed on RNA extracted from tissues of all 5 snakes. PCR amplification of samples extracted from the snake with hepatic lesions resulted in a 109-base pair (bp) product. Phylogenetic analyses indicated the virus was a novel strain distinct from other reoviruses at a level consistent with species difference. The source of infection was unknown. PCR amplification of samples extracted from the other 4 snakes was negative.


Enhancement of Radiation Sensitivity by Cathepsin L Suppression in Colon Carcinoma Cells.
Cancer is one of the main causes of death globally. Radiotherapy/Radiation therapy (RT) is one of the most common and effective cancer treatments. RT utilizes high-energy radiation to damage the DNA of cancer cells, leading to their death or impairing their proliferation. However, radiation resistance remains a significant challenge in cancer treatment, limiting its efficacy. Emerging evidence suggests that cathepsin L (cath L) contributes to radiation resistance through multiple mechanisms. In this study, we investigated the role of cath L, a member of the cysteine cathepsins (caths) in radiation sensitivity, and the potential reduction in radiation resistance by using the specific cath L inhibitor (Z-FY(tBu)DMK) or by knocking out cath L with CRISPR/Cas9 in colon carcinoma cells (caco-2). Cells were treated with different doses of radiation (2, 4, 6, 8, and 10), dose rate 3 Gy/min. In addition, the study conducted protein expression analysis by western blot and immunofluorescence assay, cytotoxicity MTT, and apoptosis assays. The results demonstrated that cath L was upregulated in response to radiation treatment, compared to non-irradiated cells. In addition, inhibiting or knocking out cath L led to increased radiosensitivity in contrast to the negative control group. This may indicate a reduced ability of cancer cells to recover from radiation-induced DNA damage, resulting in enhanced cell death. These findings highlight the possibility of targeting cath L as a therapeutic strategy to enhance the effectiveness of RT. Further studies are needed to elucidate the underlying molecular mechanisms and to assess the translational implications of cath L knockout in clinical settings. Ultimately, these findings may contribute to the development of novel treatment approaches for improving outcomes of RT in cancer patients.


Heritability jointly explained by host genotype and microbiome: will improve traits prediction?
As we observe the $70$th anniversary of the publication by Robertson that formalized the notion of 'heritability', geneticists remain puzzled by the problem of missing/hidden heritability, where heritability estimates from genome-wide association studies (GWASs) fall short of that from twin-based studies. Many possible explanations have been offered for this discrepancy, including existence of genetic variants poorly captured by existing arrays, dominance, epistasis and unaccounted-for environmental factors; albeit these remain controversial. We believe a substantial part of this problem could be solved or better understood by incorporating the host's microbiota information in the GWAS model for heritability estimation and may also increase human traits prediction for clinical utility. This is because, despite empirical observations such as (i) the intimate role of the microbiome in many complex human phenotypes, (ii) the overlap between genetic variants associated with both microbiome attributes and complex diseases and (iii) the existence of heritable bacterial taxa, current GWAS models for heritability estimate do not take into account the contributory role of the microbiome. Furthermore, heritability estimate from twin-based studies does not discern microbiome component of the observed total phenotypic variance. Here, we summarize the concept of heritability in GWAS and microbiome-wide association studies, focusing on its estimation, from a statistical genetics perspective. We then discuss a possible statistical method to incorporate the microbiome in the estimation of heritability in host GWAS.


Regulation of paraxis expression and somite formation by ectoderm- and neural tube-derived signals.
During vertebrate embryogenesis, the paraxial mesoderm becomes segmented into somites, which form as paired epithelial spheres with a periodicity that reflects the segmental organization of the embryo. As a somite matures, the ventral region gives rise to a mesenchymal cell population, the sclerotome, that forms the axial skeleton. The dorsal region of the somite remains epithelial and is called dermomyotome. The dermomyotome gives rise to the trunk and limb muscle and to the dermis of the back. Epaxial and hypaxial muscle precursors can be attributed to distinct somitic compartments which are laid down prior to overt somite differentiation. Inductive signals from the neural tube, notochord, and overlying ectoderm have been shown to be required for patterning of the somites into these different compartments. Paraxis is a basic helix-loop-helix transcription factor expressed in the unsegmented paraxial mesoderm and throughout epithelial somites before becoming restricted to epithelial cells of the dermomyotome. To determine whether paraxis might be a target for inductive signals that influence somite patterning, we examined the influence of axial structures and surface ectoderm on paraxis expression by performing microsurgical operations on chick embryos. These studies revealed two distinct phases of paraxis expression, an early phase in the paraxial mesoderm that is dependent on signals from the ectoderm and independent of the neural tube, and a later phase that is supported by redundant signals from the ectoderm and neural tube. Under experimental conditions in which paraxis failed to be expressed, cells from the paraxial mesoderm failed to epithelialize and somites were not formed. We also performed an RT-PCR analysis of combined tissue explants in vitro and confirmed that surface ectoderm is sufficient to induce paraxis expression in segmental plate mesoderm. These results demonstrate that somite formation requires signals from adjacent cell types and that the paraxis gene is a target for the signal transduction pathways that regulate somitogenesis.


Perturbation-Based Balance Exercise Using a Wearable Device to Improve Reactive Postural Control.
Reactive postural control is an important component of the balance function for fall prevention. Perturbation-based balance exercises improve reactive postural control; however, these exercises require large, complex instruments and expert medical guidance. This study investigates the effects of unexpected perturbation-based balance exercises using a wearable balance exercise device (WBED) on reactive postural control. Eighteen healthy adult males participated in this study. Participants were assigned to the WBED and Sham groups. In the intervention session, participants in the WBED group randomly underwent unexpected perturbation in the mediolateral direction, while the Sham group performed the same exercises without perturbation. Before and after the intervention session, all participants underwent evaluation of reactive balance function using air cylinders. Peak displacement (D), time at peak displacement (T), peak velocity (V), and root mean square (RMS) of center of pressure (COP) data were measured. For mediolateral and anteroposterior COP (COPML and COP[Formula: see text]), the main effects of group and time factors (pre/post) were investigated through the analysis of variance for split-plot factorial design. In the WBED group, the D-COPML and V-COPML of the post-test significantly decreased compared to those of the pre-test (p = 0.017 and p = 0.003, respectively). Furthermore, the D-COPAP and RMSAP of the post-test significantly decreased compared to those of the pre-test (p = 0.036 and p = 0.015, respectively). This study proved that the perturbation-based balance exercise using WBED immediately improved reactive postural control. Therefore, wearable exercise devices, such as WBED, may contribute to the prevention of falls and fall-related injuries.


SSN20 is an essential gene with mutant alleles that suppress defects in SUC2 transcription in Saccharomyces cerevisiae.
Dominant and recessive mutations at the SSN20 locus were previously isolated as extragenic suppressors of mutations in three genes (SNF2, SNF5, and SNF6) that are required in trans to derepress invertase expression. All ssn20 alleles cause recessive, temperature-sensitive lethality. In this study we cloned the SSN20 gene, identified a 4.6-kilobase poly(A)-containing RNA, and showed that disruption of the gene is lethal in a haploid cell. Genetic mapping of SSN20 to a locus on chromosome VII 10 centimorgans distal to cly8 led to the finding that SSN20 is the same gene as SPT6, which affects expression of delta insertions in the 5' noncoding region of HIS4 (F. Winston, D. T. Chaleff, B. Valent, and G. R. Fink, Genetics 107:179-197, 1984). We also showed that an ssn20 mutation restored expression of secreted invertase from deletions of the SUC2 upstream regulatory region; ssn20 restored derepression of SUC2 mRNA in strains with a SUC2 upstream region deletion or a snf2 mutation. Increased or decreased gene dosage of SSN20 also suppressed defects that are suppressed by ssn20 missense mutations. These findings suggest that SSN20 plays a role in general transcriptional processes.


Metabolic engineering for the production of natural products.
Natural products and their derivatives play an important role in modern healthcare as frontline treatments for many diseases and as inspiration for chemically synthesized therapeutics. With advances in sequencing and recombinant DNA technology, many of the biosynthetic pathways responsible for the production of these chemically complex yet valuable compounds have been elucidated. With an ever-expanding toolkit of biosynthetic components, metabolic engineering is an increasingly powerful method to improve natural product titers and generate novel compounds. Heterologous production platforms have enabled access to pathways from difficult to culture strains, systems biology and metabolic modeling tools have resulted in increasing predictive and analytic capabilities, advances in expression systems and regulation have enabled the fine-tuning of pathways for increased efficiency, and characterization of individual pathway components has facilitated the construction of hybrid pathways for the production of new compounds. These advances in the many aspects of metabolic engineering not only have yielded fascinating scientific discoveries but also make it an increasingly viable approach for the optimization of natural product biosynthesis.


Is there benefit in dual renin-angiotensin-aldosterone system blockade? No, yes and maybe: a guide for the perplexed.
Since the initial discovery of Angiotensin converting enzyme inhibitors (ACEI) in the 1960s and the launch of Captopril as the first available for clinical use in the 1970s, there now exist three other classes of drugs that block the renin angiotensin aldosterone system (RAAS): the angiotensin II receptor blockers (ARB), aldosterone antagonists (AA) and direct renin inhibitors (DRI). With the proven efficacy of RAAS blockers as monotherapy in many arenas there has been considerable interest in the use of dual therapy combinations of these medications that target different points in the pathway. By potentially offering a more complete RAAS blockade with a commensurate enhanced clinical effect, the strong biological rationale for dual therapy has led to it being embraced by clinicians as a treatment option, for hypertension and nephroprotection in particular. However, the initial enthusiasm for this treatment has been tempered by the recent results from several large trials such as ONTARGET and ALTITUDE, which do not support a specific dual therapy approach. In contrast, there is supportive evidence for dual blockade of specific combinations in selected patient groups and data are lacking for others. In the wake of this complex contemporary evidence, the conundrum now faced by clinicians committed to individualised care is, for which patients dual therapy could still be of benefit. This review examines for the practising clinician the current 'state of play' for dual blockade of various combinations and a perspective on its use in cardio-renal disease and diabetic complications.


Development of cytochrome oxidase blobs in visual cortex of normal and visually deprived cats.
Cytochrome oxidase (CO) blobs are central to our understanding of the columnar organization and parallel pathways in primate and cat visual cortex. In primates, development of blobs and their relationship with other columnar features of the visual cortex begins pre-natally, before visual experience. In kittens, the supragranular layers differentiate post-natally, after eye opening, raising the possibility that visual experience may influence the development of blobs in cat V1. We have examined the development of blobs in unfolded and flattened sections through the visual cortex of normally reared, dark-reared, monocularly deprived and binocularly deprived kittens. Blobs were found in superficial layers of V1 of normally reared kittens as early as 2 weeks of age, although at this age the overall CO staining in V1 was lighter than in V2. By 6 weeks of age the blobs were adult-like. A patchy pattern of CO staining was also found in V2 of young kittens but not in adults. Visual experience was not necessary for expression of the blobs and monocularly deprived kittens had well developed blobs, indicating that strong Y cell drive is not necessary for the development of blobs in cat V1. CO blobs appear in kitten V1 very early in post-natal development and their expression is independent of visual experience, suggesting that they may be an intrinsic feature of V1 organization.


Assessment of pharmaceutical waste management at selected hospitals and homes in Ghana.
The practice of use and disposal of waste from pharmaceuticals compromises the safety of the environment as well as representing a serious health risk, as they may accumulate and stay active for a long time in the aquatic environment. This article therefore presents the outcome of a study on pharmaceutical waste management practices at homes and hospitals in Ghana. The study was conducted at five healthcare institutions randomly selected in Ghana, namely two teaching hospitals (hospital A, hospital B), one regional hospital (hospital C), one district hospital (hospital D) and one quasi-governmental hospital (hospital E). Apart from hospital E which currently has a pharmaceutical waste separation programmr as well as drug return programme called DUMP (Disposal of Unused Medicines Program), all other hospitals visited do not have any separate collection and disposal programme for pharmaceutical waste. A survey was also carried out among the general public, involving the questioning of randomly selected participants in order to investigate the household disposal of unused and expired pharmaceuticals. The results from the survey showed that more than half of the respondents confirmed having unused, left-over or expired medicines at home and over 75% disposed of pharmaceutical waste through the normal waste bins which end up in the landfills or dump sites.


Conformationally-constrained indeno[2,1-c]quinolines--a new class of anti-mycobacterial agents.
The design, synthesis and anti-mycobacterial activities of 23 conformationally-constrained indeno[2,1-c]quinolines against Mycobacterium tuberculosis H37Rv is reported. Based on a structural comparison with the anti-TB TMC207 we have devised a synthetic methodology for making new conformationally-constrained indeno[2,1-c]quinoline analogs (Fig. 1), by retaining the biologically significant quinoline and the phenyl rings in the SW and NW hemispheres, respectively. This new class of conformationally-constrained compounds has been designed such that their conformational flexibility across C4-C2' is diminished to nil by covalently locking the C4 center of the quinoline moiety in the SW hemisphere with the C2' center of the phenyl ring in the NW hemisphere, thereby decreasing the entropic penalty for their complex formation within the target protein, which will in turn give improved free-energy of stabilization of the complex. The efficacies of these anti-TB compounds were evaluated in vitro for 8/9 consecutive days using the BACTEC radiometric assay upon administration of a single-dose on day one. Compounds 11, 13, 16, 24, 30, 32 and 34 showed 85-99% growth inhibition of Mycobacterium tuberculosis. Compounds 13 and 34 however have inhibited the mycobacterial growth more effectively than others in the series, with minimum inhibitory concentrations (MIC) of 0.39 microg mL(-1) (1 microM) and 0.78 microg mL(-1) (2 microM) respectively.


Endothelial dysfunction in pre-eclampsia.
The repeated demonstration of biomarkers of endothelial cell and leukocyte activation has suggested that the maternal syndrome of pre-eclampsia arises from a generalised maternal inflammatory systemic response incorporating a substantive component of endothelial cell dysfunction. Reports of reduced endothelium dependent dilatation in isolated resistance arteries and from non invasive methods in vivo indicate a major contribution to the systemic vasoconstriction, characteristic of the syndrome. The recent discovery of raised concentrations of soluble fms-like tyrosine kinase1(sFlt1) and the soluble transforming growth factor beta (TGF-beta) coreceptor (sEng) sEng which indirectly may compromise endothelial function, adds to the growing list of potential origins of endothelial disturbance. Most are proposed to originate from placental underperfusion and associated placental oxidative stress, although it is clear that not all women with pre-eclampsia have reduced utero-placental blood flow, and other precipitating factors, including dyslipidaemia and hyperglycaemia are likely to contribute. Endothelial dysfunction, unlike pre-eclampsia, does not resolve post-partum, and persistence of the defect may underpin the increased risk of cardiovascular disease in later life.


Caloribacterium cisternae gen. nov., sp. nov., an anaerobic thermophilic bacterium from an underground gas storage reservoir.
A novel anaerobic, moderately thermophilic bacterium (strain SGL43(T)) was isolated from Severo-Stavropolskoye underground gas storage reservoir (Russia). Cells of strain SGL43(T) were motile straight rods, 0.4 µm in diameter and 2.0-3.0 µm in length. The temperature range for growth was 28-65 °C, with optimum growth at 50 °C. The pH range for growth was 5.5-8.0, with optimum growth at pH 7.0-7.5. Growth of strain SGL43(T) was observed at NaCl concentrations of 0-4.0% (w/v) with optimum growth at 1.0% (w/v) NaCl. Substrates utilized by strain SGL43(T) included peptone, yeast extract, glucose, fructose, maltose, galactose, pyruvate and citrate. Products of glucose or citrate fermentation were acetate, hydrogen and CO(2). Thiosulfate was reduced to sulfide. The DNA G+C content of strain SGL43(T) was 43.1 mol%. 16S rRNA gene sequence analysis revealed that strain SGL43(T) belongs to the order Thermoanaerobacterales (phylum 'Firmicutes'). The closest relative of strain SGL43(T) was Thermoanaerobacterium saccharolyticum (86.2% 16S rRNA gene sequence similarity with the type strain). Based on the data presented here, strain SGL43(T) is considered to represent a novel species of a new genus, for which the name Caloribacterium cisternae gen. nov., sp. nov. is proposed. The type strain of Caloribacterium cisternae, the type species of the genus, is SGL43(T) (=DSM 23830(T)=VKM B-2670(T)).


MotivATE: A Pretreatment Web-Based Program to Improve Attendance at UK Outpatient Services Among Adults With Eating Disorders.
In the UK, eating disorders affect upward of 725,000 people per year, and early assessment and treatment are important for patient outcomes. Around a third of adult outpatients in the UK who are referred to specialist eating disorder services do not attend, which could be related to patient factors related to ambivalence, fear, and a lack of confidence about change. This lack of engagement has a negative impact on the quality of life of patients and has implications for service costs. To describe the development of a Web-based program ("MotivATE") designed for delivery at the point of referral to an eating disorder service, with the aim of increasing service attendance. We used intervention mapping and a person-based approach to design the MotivATE program and conducted a needs assessment to determine the current impact of service nonattendance on patients (via a review of the qualitative evidence) and services (through a service provision survey to understand current issues in UK services). Following the needs assessment, we followed the five steps of program development outlined by Bartholomew et al (1998): (1) creating a matrix of proximal program objectives; (2) selecting theory-based intervention methods and strategies; (3) designing and organizing the program; (4) specifying adoption and implementation plans; and (5) generating program evaluation plans. The needs assessment identified current nonattendance rates of 10%-32%. We defined the objective of MotivATE as increasing attendance rates at an eating disorder service and considered four key determinants of poor attendance: patient ambivalence about change, low patient self-efficacy, recognition of the need to change, and expectations about assessment. We chose aspects of motivational interviewing, self-determination theory, and the use of patient stories as the most appropriate ways to enable change. Think-aloud piloting with people with lived experience of an eating disorder resulted in positive feedback on the MotivATE program. Participants related well to the stories used. Nonetheless, because of feedback, we further modified the program in line with patients' stage of change and addressed issues with the language used. A consultation with service staff meant that we could make clear implementation plans. Finally, a randomized controlled trial is currently underway to evaluate the MotivATE program. Using intervention mapping, we have developed a novel pretreatment Web-based program that is acceptable to people with eating disorders. To our knowledge, this is the first such program. The model of development described here could be a useful template for designing further programs for other difficult-to-engage populations.


Unexpected pulmonary involvement in extrapulmonary tuberculosis patients.
This study aimed to assess the utility of sputum examinations and chest radiographs (CXRs) in patients with extrapulmonary tuberculosis (XPTB) to detect pulmonary involvement of tuberculosis (TB). We studied 72 XPTB patients who were managed through the TB Program, King County, WA, from January 2003 through November 2004. The two most common sites of XPTB were the lymph nodes (36 [50%]) and pleura (12 [17%]). Thirty-five of 72 XPTB patients (49%) had abnormal CXR findings. Sputum was not obtained from 15 patients despite sputum induction. Of the 57 patients from whom sputum was collected, 30 (53%) had abnormal CXR findings, 5 (9%) had sputum smears that were positive for acid-fast bacilli, and 12 (21%) had sputum cultures that were positive for Mycobacterium tuberculosis. Weight loss was significantly associated with positive sputum culture findings in a multivariate analysis (odds ratio, 4.3; 95% confidence interval, 1.01 to 18.72; p = 0.049). There was no significant difference in the occurrence of positive sputum culture results between patients with abnormal CXR findings and those with normal CXR findings (7 of 30 patients [23%] vs 5 of 27 patients [19%], respectively; p = 0.656). Of 24 HIV-negative XPTB patients with normal CXR findings, 2 patients (8%) had positive sputum culture findings. CXR results did not reliably differentiate XPTB patients with and without positive sputum culture findings. Some XPTB patients had positive sputum culture results despite normal CXR findings and negative HIV status. Weight loss in XPTB patients was associated with positive sputum culture results. Sputum examinations in XPTB patients, regardless of the CXR results, may identify potentially infectious cases of TB.