Heterologous, Fresh, Human Donor Sclera as Patch Graft Material in Glaucoma Drainage Device Surgery.
To determine the safety and efficacy of fresh, human sclera allografts as a patch graft material in glaucoma drainage device (GDD) surgery. Retrospective, noncomparative, interventional, consecutive case series. All GDD cases operated between 2008 and 2013 in which fresh human corneoscleral rims were used immediately after the central corneal button was used for penetrating or endothelial keratoplasty. Surgery was performed by 2 surgeons at 2 facilities. The Ahmed Glaucoma Valve (FP-7) was used exclusively in this cohort. Sixty-four eyes of 60 patients were identified; demographic data were recorded along with intraocular pressure (IOP), medication requirements, visual acuity, complications, and subsequent interventions. Incidence of complications. IOP and medication requirements at the last follow-up. Quilified success utilizing Tube Versus Trabeculectomy study criteria. The mean age of the cohort was 66.2±19.1 years; the average preoperative IOP was 33.2±11.1 mm Hg on 4.2±1.3 IOP-lowering agents before GDD surgery. IOP decreased significantly to 14.1±4.7 mm Hg (P<0.001) on 1.6±1.2 IOP-lowering agents (P<0.001) after an average follow-up of 18.2±15.4 months. There were no cases of early or late blebitis or endophthalmitis, and there was 1 case of conjunctival erosion and tube/plate exposure (1.6%) occurring 30 days after surgery. Qualified success was estimated as 90.5% and 81% at 1 and 2 years, respectively, using Tube Versus Trabeculectomy study criteria. Heterologous, fresh, human donor sclera appears to be a safe material for GDD tube coverage. It provides a cost-efficient alternative compared with traditional patch graft materials associated with a low risk of pathogen transmission.


Interleukin-6, produced by resident cells of the central nervous system and infiltrating cells, contributes to the development of seizures following viral infection.
Cells that can participate in an innate immune response within the central nervous system (CNS) include infiltrating cells (polymorphonuclear leukocytes [PMNs], macrophages, and natural killer [NK] cells) and resident cells (microglia and sometimes astrocytes). The proinflammatory cytokine interleukin-6 (IL-6) is produced by all of these cells and has been implicated in the development of behavioral seizures in the Theiler's murine encephalomyelitis virus (TMEV)-induced seizure model. The assessment, via PCR arrays, of the mRNA expression levels of a large number of chemokines (ligands and receptors) in TMEV-infected and mock-infected C57BL/6 mice both with and without seizures did not clearly demonstrate the involvement of PMNs, monocytes/macrophages, or NK cells in the development of seizures, possibly due to overlapping function of the chemokines. Additionally, C57BL/6 mice unable to recruit or depleted of infiltrating PMNs and NK cells had seizure rates comparable to those of controls following TMEV infection, and therefore PMNs and NK cells do not significantly contribute to seizure development. In contrast, C57BL/6 mice treated with minocycline, which affects monocytes/macrophages, microglial cells, and PMNs, had significantly fewer seizures than controls following TMEV infection, indicating monocytes/macrophages and resident microglial cells are important in seizure development. Irradiated bone marrow chimeric mice that were either IL-6-deficient mice reconstituted with wild-type bone marrow cells or wild-type mice reconstituted with IL-6-deficient bone marrow cells developed significantly fewer behavioral seizures following TMEV infection. Therefore, both resident CNS cells and infiltrating cells are necessary for seizure development.


Reliability measures in distorting body-image.
This study is the first to make a direct comparison between the distorting mirror and the distorting videocamera with the same group of nonclinical subjects. It also establishes the internal consistency, the test-retest reliability and the convergent validity of both techniques. Analysis indicates, as with other studies, that perceived image is consistently over-estimated and ideal image is consistently under-estimated by both techniques. With the distorting mirror, the frontal orientation is repeatable over four days and with the video the profile is the more reliable orientation. A comparison between profile and frontal orientations on any single test occasion (internal consistency) indicates that the results correlate well so either orientation is acceptable for experimental purposes. The extent to which both techniques were measuring the same aspect (convergent validity) was more marked for the ideal image. A researcher using the distorting mirror can be confident that the ideal image is reliable over time using either orientation but that the perceived image requires a frontal orientation. The frontal or profile orientation is acceptable for the ideal image using the distorting video, but the profile orientation is advisable if one wishes a reliable perceived image. The researcher should also be cautious in assuming that both techniques are measuring an identical body-image construct.


Rice P1B-type heavy-metal ATPase, OsHMA9, is a metal efflux protein.
P(1B)-type heavy-metal ATPases (HMAs) are transmembrane metal-transporting proteins that play a key role in metal homeostasis. Despite their importance, very little is known about their functions in monocot species. We report the characterization of rice (Oryza sativa) OsHMA9, a member of the P(1B)-type ATPase family. Semiquantitative reverse transcription-polymerase chain reaction analyses of seedlings showed that OsHMA9 expression was induced by a high concentration of copper (Cu), zinc (Zn), and cadmium. We also determined, through promoterbeta-glucuronidase analysis, that the main expression was in the vascular bundles and anthers. The OsHMA9:green fluorescence protein fusion was localized to the plasma membrane. Heterologous expression of OsHMA9 partially rescued the Cu sensitivity of the Escherichia coli copA mutant, which is defective in Cu-transporting ATPases. It did not rescue the Zn sensitivity of the zntA mutant, which is defective in Zn-transporting ATPase. To further elucidate the functional roles of OsHMA9, we isolated two independent null alleles, oshma9-1 and oshma9-2, from the T-DNA insertion population. Mutant plants exhibited the phenotype of increased sensitivity to elevated levels of Cu, Zn, and lead. These results support a role for OsHMA9 in Cu, Zn, and lead efflux from the cells. This article is the first report on the functional characterization of a P(1B)-type metal efflux transporter in monocots.


A hypertension risk score for adults: a population-based cross-sectional study from the Dubai Household Survey 2019.
The aim of this study was to develop a risk score model for predicting hypertension specific to the population of Dubai in the United Arab Emirates (UAE) to facilitate prevention and early intervention. A retrospective analysis of data from the Dubai Household Health Survey 2019 was conducted. Demographic and physical parameters, as well as blood glucose levels, were included in the data. The risk factors for hypertension were identified using bivariate analysis. A risk score model was developed using the enter method, where all significant predictors of hypertension in bivariate analyses were entered in a single step with the primary outcome of hypertension status (yes/no). The model was validated internally by splitting the data into Emirati and non-Emirati populations. A total of 2,533 subjects were studied. The significant risk factors for hypertension identified were male sex, older age (≥40 years), education level, body mass index, diabetes mellitus, and dyslipidaemia. The model showed a high discrimination ability between individuals with and without hypertension, with an area under the curve of 0.77 (95% confidence interval [CI], 0.75 to 0.79), excellent sensitivity (81.0%; 95% CI, 71.9 to 88.2) and moderate specificity (56.0%; 95% CI, 45.7 to 65.9). The model developed by this study is simple, convenient, and based on readily available demographic and medical characteristics. This risk score model could support initial hypertension screening and provide an effective tool for targeted lifestyle counselling and prevention programs.


Physiologic approach for coronary intervention.
When invasively assessing coronary artery disease, the primary goal should be to determine whether the disease is causing a patient's symptoms and whether it is likely to cause future cardiac events. The presence of myocardial ischemia is our best gauge of whether a lesion is responsible for symptoms and likely to result in a future cardiac event. In the catheterization laboratory, fractional flow reserve (FFR) measured with a coronary pressure wire is the reference standard for identifying ischemia-producing lesions. Its spatial resolution is unsurpassed with it not only being vessel-specific, but also lesion-specific. There is now a wealth of data supporting the accuracy of measuring FFR to identify ischemia-producing lesions. FFR-guided percutaneous coronary intervention of these lesions results in improved outcomes and saves resources. Non-hemodynamically significant lesions can be safely managed medically with a low rate of subsequent cardiac events.


Confirmed testicular torsion in a 67 year old.
Torsion amongst the elderly population is rarely described. This case presents the oldest surgically confirmed case of testicular torsion, in a 67-year-old male, within the UK. Presenting to the emergency department with a 10-day history of left-sided testicular pain, initially treated with antibiotics. There was no pyrexia or urinary symptoms and negative urine dipstick. In adults above the age of 40, likely diagnoses include epididymo-orchitis, epididymitis, neoplasm or hydrocele. Clinical differentiation with epididymo-orchitis can be difficult in any age range. Clinical signs such as fever, elevated C-reactive protein and positive urine dipstick test are suggestive of epididymo-orchitis/orchitis. This case study demonstrates that testicular torsion can occur at any age, and clinical suspicion should always be high in patients presenting with testicular pain and a negative urine dipstick, regardless of age. Although risk in this subgroup is low, the identification of a potentially reversible testicular abnormality should be of high priority.


Paternal imprinting in Marchantia polymorpha.
We are becoming aware of a growing number of organisms that do not express genetic information equally from both parents as a result of an epigenetic phenomenon called genomic imprinting. Recently, it was shown that the entire paternal genome is repressed during the diploid phase of the life cycle of the liverwort Marchantia polymorpha. The deposition of the repressive epigenetic mark H3K27me3 on the male pronucleus is responsible for the imprinted state, which is reset by the end of meiosis. Here, we put these recent reports in perspective of other forms of imprinting and discuss the potential mechanisms of imprinting in bryophytes and the causes of its evolution.


Salivary agglutinin, which binds Streptococcus mutans and Helicobacter pylori, is the lung scavenger receptor cysteine-rich protein gp-340.
Salivary agglutinin is a high molecular mass component of human saliva that binds Streptococcus mutans, an oral bacterium implicated in dental caries. To study its protein sequence, we isolated the agglutinin from human parotid saliva. After trypsin digestion, a portion was analyzed by matrix-assisted laser/desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), which gave the molecular mass of 14 unique peptides. The remainder of the digest was subjected to high performance liquid chromatography, and the separated peptides were analyzed by MALDI-TOF/post-source decay; the spectra gave the sequences of five peptides. The molecular mass and peptide sequence information showed that salivary agglutinin peptides were identical to sequences in lung (lavage) gp-340, a member of the scavenger receptor cysteine-rich protein family. Immunoblotting with antibodies that specifically recognized either lung gp-340 or the agglutinin confirmed that the salivary agglutinin was gp-340. Immunoblotting with an antibody specific to the sialyl Le(x) carbohydrate epitope detected expression on the salivary but not the lung glycoprotein, possible evidence of different glycoforms. The salivary agglutinin also interacted with Helicobacter pylori, implicated in gastritis and peptic ulcer disease, Streptococcus agalactiae, implicated in neonatal meningitis, and several oral commensal streptococci. These results identify the salivary agglutinin as gp-340 and suggest it binds bacteria that are important determinants of either the oral ecology or systemic diseases.


Nonpigmenting solitary fixed drug eruption caused by a Chinese traditional herbal medicine, ma huang (Ephedra Hebra), mainly containing pseudoephedrine and ephedrine.
We describe a case of nonpigmenting solitary fixed drug eruption appearing on the right thigh of a 31-year-old woman in Japan. The causative drug was determined by closed patch test on postlesional skin as a Chinese traditional herbal medicine, ma huang (Ephedra Hebra), mainly containing pseudoephedrine and ephedrine.


Microfluidic glucose stimulation reveals limited coordination of intracellular Ca2+ activity oscillations in pancreatic islets.
The pancreatic islet is a functional microorgan involved in maintaining normoglycemia through regulated secretion of insulin and other hormones. Extracellular glucose stimulates insulin secretion from islet beta cells through an increase in redox state, which can be measured by NAD(P)H autofluorescence. Glucose concentrations over approximately 7 mM generate synchronous oscillations in beta cell intracellular Ca2+ concentration ([Ca2+]i), which lead to pulsatile insulin secretion. Prevailing models assume that the pancreatic islet acts as a functional syncytium, and the whole islet [Ca2+]i response has been modeled in terms of islet bursting and pacemaker models. To test these models, we developed a microfluidic device capable of partially stimulating an islet, while allowing observation of the NAD(P)H and [Ca2+]i responses. We show that beta cell [Ca2+]i oscillations occur only within regions stimulated with more than approximately 6.6 mM glucose. Furthermore, we show that tolbutamide, an antagonist of the ATP-sensitive K+ channel, allows these oscillations to travel farther into the nonstimulated regions of the islet. Our approach shows that the extent of Ca2+ propagation across the islet depends on a delicate interaction between the degree of coupling and the extent of ATP-sensitive K+-channel activation and illustrates an experimental paradigm that will have utility for many other biological systems.


Eugenol as a Promising Molecule for the Treatment of Dermatitis: Antioxidant and Anti-inflammatory Activities and Its Nanoformulation.
Contact dermatitis produces an inflammatory reaction primarily via stimulation of keratinocytes and cells of the immune system, which promote the release of cytokines, reactive oxygen species (ROS), and other chemical mediators. Eugenol (EUG, phenylpropanoid of essential oils) has attracted attention due to its anti-inflammatory properties, as well as antioxidant effect. On the other hand, it is volatile and insoluble and is a skin irritant. In this case, nanostructured systems have been successfully employed as a drug carrier for skin diseases since they improve both biological and pharmaceutical properties of active compounds. The cytotoxic, antioxidant, and anti-inflammatory effects of EUG were assessed in human neutrophils and keratinocytes. Additionally, polymeric nanocarries (NCEUG) were prepared to improve the chemical and irritant characteristics of EUG. EUG presented apparent safety and antioxidant and anti-inflammatory effects on human neutrophils, but presented cytotoxic effects on keratinocytes. However, the nanocapsules were able to reduce its cytotoxicity. An in vivo experiment of irritant contact dermatitis (ICD) in mice induced by TPA showed that NCEUG reduced significantly the ear edema in mice when compared to the EUG solution, as well as the leukocyte infiltration and IL-6 level, possibly due to better skin permeation and irritancy blockage. These findings suggest that EUG is a promising bioactive molecule, and its nanoencapsulation seems to be an interesting approach for the treatment of ICD.


Extending gene ontology in the context of extracellular RNA and vesicle communication.
To address the lack of standard terminology to describe extracellular RNA (exRNA) data/metadata, we have launched an inter-community effort to extend the Gene Ontology (GO) with subcellular structure concepts relevant to the exRNA domain. By extending GO in this manner, the exRNA data/metadata will be more easily annotated and queried because it will be based on a shared set of terms and relationships relevant to extracellular research. By following a consensus-building process, we have worked with several academic societies/consortia, including ERCC, ISEV, and ASEMV, to identify and approve a set of exRNA and extracellular vesicle-related terms and relationships that have been incorporated into GO. In addition, we have initiated an ongoing process of extractions of gene product annotations associated with these terms from Vesiclepedia and ExoCarta, conversion of the extracted annotations to Gene Association File (GAF) format for batch submission to GO, and curation of the submitted annotations by the GO Consortium. As a use case, we have incorporated some of the GO terms into annotations of samples from the exRNA Atlas and implemented a faceted search interface based on such annotations. We have added 7 new terms and modified 9 existing terms (along with their synonyms and relationships) to GO. Additionally, 18,695 unique coding gene products (mRNAs and proteins) and 963 unique non-coding gene products (ncRNAs) which are associated with the terms: "extracellular vesicle", "extracellular exosome", "apoptotic body", and "microvesicle" were extracted from ExoCarta and Vesiclepedia. These annotations are currently being processed for submission to GO. As an inter-community effort, we have made a substantial update to GO in the exRNA context. We have also demonstrated the utility of some of the new GO terms for sample annotation and metadata search.


Abscess Incision and Drainage With or Without Ultrasonography: A Randomized Controlled Trial.
We hypothesize that clinical failure rates will be lower in patients treated with point-of-care ultrasonography and incision and drainage compared with those who undergo incision and drainage after physical examination alone. We performed a prospective randomized clinical trial of patients presenting with a soft tissue abscess at a large, academic emergency department. Patients presenting with an uncomplicated soft tissue abscess requiring incision and drainage were eligible for enrollment and randomized to treatment with or without point-of-care ultrasonography. The diagnosis of an abscess was by physical examination, bedside ultrasonography, or both. Patients randomized to the point-of-care ultrasonography group had an incision and drainage performed with bedside ultrasonographic imaging of the abscess. Patients randomized to the non-point-of-care ultrasonography group had an incision and drainage performed with physical examination alone. Comparison between groups was by comparing means with 95% confidence intervals. The primary outcome was failure of therapy at 10 days, defined as a repeated incision and drainage, following a per-protocol analysis. Multivariate analysis was performed to control for study variables. Our study was designed to detect a clinically important difference between groups, which we defined as a 13% difference. A total of 125 patients were enrolled, 63 randomized to the point-of-care ultrasonography group and 62 to physical examination alone. After loss to follow-up and misallocation, 54 patients in the ultrasonography group and 53 in the physical examination alone group were analyzed. The overall failure rate for all patients enrolled in the study was 10.3%. Patients who were evaluated with ultrasonography were less likely to fail therapy and have repeated incision and drainage, with a difference between groups of 13.3% (95% confidence interval 0.0% to 19.4%). Abscess locations were predominantly torso (21%), buttocks (21%), lower extremity (18%), and axilla or groin (16%). There was no difference in baseline characteristics between groups relative to abscess size, duration of symptoms before presentation, percentage with cellulitis, and treatment with antibiotics. Patients with soft tissue abscesses who were undergoing incision and drainage with point-of-care ultrasonography demonstrated less clinical failure compared with those treated without point-of-care ultrasonography.


Magnetic field-dependent resistance crossover and anomalous magnetoresistance in topological insulator Bi2Te3.
We report a metal-insulator like transition in single-crystalline 3D topological insulator Bi2Te3 at a temperature of 230 K in the presence of an external magnetic field applied normal to the surface. This transition becomes more prominent at larger magnetic field strength with the residual resistance value increasing linearly with the magnetic field. At low temperature, the magnetic field dependence of the magnetoresistance shows a transition from logarithmic to linear behavior and the onset magnetic field value for this transition decreases with increasing temperature. The logarithmic magnetoresistance indicates the weak anti-localization of the surface Dirac electrons while the high temperature behavior originates from the bulk carriers due to intrinsic impurities. At even higher temperatures beyond ∼230 K, a completely classical Lorentz model type quadratic behavior of the magnetoresistance is observed. We also show that the experimentally observed anomalies at ∼230 K in the magneto-transport properties do not originate from any stacking fault in Bi2Te3.


Separation of full and empty adeno-associated virus capsids by anion-exchange chromatography using choline-type salts.
Development of clinically desirable adeno-associated virus (AAV) vectors with optimal genome design requires rapid and accurate analytical methods to assess AAV quality. Anion-exchange (AEX) chromatography provides a powerful analytical method for full/empty AAV capsid ratio determination. However, the current AEX methodology for separation of empty and full AAV capsids largely relies on the use of the highly toxic tetramethylammonium chloride (TMAC). Here, we describe a novel analytical AEX method for separation of empty and full AAV capsids that uses only non-toxic, choline-type compounds that contain structural similarity to the quaternary ammonium ligand present on the surface of AEX resin. Choline-Cl gradient, combined with sensitive fluorescence detection, allowed a safe and effective separation of empty and full AAV capsids with reproducible empty/full ratio determination. The choline-based assay was suitable for commonly used serotypes, AAV2, AAV5, AAV6, and AAV8. The limit of detection was ∼3.9 × 108 virus particles in the assay. A gradient-hold step-gradient elution with choline-Cl resulted in enhanced baseline separation of empty and full AAV8 capsids. In summary, the use of choline-Cl in the AEX assay is recommended for empty/full capsid ratio determination and other applications in AAV production, and it eliminates the necessity of using toxic TMAC.


Morpho-chemical characterization of individual ancient starches retrieved on ground stone tools from Palaeolithic sites in the Pontic steppe.
Despite the extensive literature on the retrieval of digestible starches from archaeological contexts, there are still significant concerns regarding their genuine origin and durability. Here, we propose a multi-analytical strategy to identify the authenticity of ancient starches retrieved from macrolithic tools excavated at Upper Paleolithic sites in the Pontic steppe. This strategy integrates the morphological discrimination of starches through optical microscopy and scanning electron microscopy with single starch chemo-profiling using Fourier transform infrared imaging and microscopy. We obtained evidence of aging and biomineralization in the use-related starches from Palaeolithic sites, providing a methodology to establish their ancient origin, assess their preservation status, and attempt their identification. The pivotal application of this multidisciplinar approach demonstrates that the macrolithic tools, from which starches were dislodged, were used for food-processing across the Pontic Steppe around 40,000 years ago during the earliest colonization of Eurasia by Homo sapiens.


Prenatal diagnosis of musculoskeletal anomalies.
Musculoskeletal anomalies are not uncommon in prenatal life. They can be either sporadic or part of chromosomal syndromes causing prenatal morbidity and mortality. The prenatal diagnosis of musculoskeletal anomalies is based on information assembled from various imaging modalities and from biochemical and genetic workups. The prenatal diagnosis can serve as a prognostic tool and in counseling the parents. Among the imaging methods, ultrasonography is the most popular and cost effective in observing and following fetal development from the very early stages of gestation. Transvaginal sonography can detect and identify most of the normal and the specific pathologic changes very close to the stage of their embryogenic development. From a practical point of view, early detailed transvaginal sonography screening at 14 to 15 weeks of gestation is very useful while late detection at 20 to 23 weeks of gestation may provide some additional information in low-risk pregnancies. Very early screening, even during the ninth week, may be indicated in high-risk pregnancies. Additional genetic counseling is recommended when abnormal findings are suspected. We summarize the diagnostic approach and the information available for the most common musculoskeletal anomalies.


Symmetry of foot alignment and ankle flexibility in paediatric Charcot-Marie-Tooth disease.
Charcot-Marie-Tooth disease is the most common inherited nerve disorder and typically presents with pes cavus foot deformity and ankle equinus during childhood. Level in the variation of symmetry of musculoskeletal lower limb involvement across the clinical population is unknown, despite early reports describing gross asymmetry. We measured foot alignment and ankle flexibility of the left and right limbs using accurate and reliable standardised paediatric outcome measures in 172 patients aged 3-20 years with a variety of disease subtypes recruited from the United States, United Kingdom, Italy and Australia. While a large range of differences existed between left and right feet for a small proportion of children, there was no overall significant difference between limbs. There are two important implications of these findings. Children with Charcot-Marie-Tooth disease generally exhibit symmetrical foot alignment and ankle flexibility between limbs. As such, analysing one limb only for biomechanical-related research is appropriate and satisfies the independence requirements for statistical analysis. However, because there are large differences between feet for a small proportion of children, an individualised limb-focused approach to clinical care is required.


Diabetic neuropathic pain: a role for testosterone metabolites.
Diabetic neuropathy is associated with neuropathic pain in about 50% of diabetic subjects. Clinical management of neuropathic pain is complex and so far unsatisfactory. In this study, we analyzed the effects of the testosterone metabolites, dihydrotestosterone (DHT), and 3α-diol, on nociceptive and allodynia thresholds and on molecular and functional parameters related to pain modulation in the dorsal horns of the spinal cord and in the dorsal root ganglia of rats rendered diabetic by streptozotocin injection. Furthermore, the levels of DHT and 3α-diol were analyzed in the spinal cord. Diabetes resulted in a significant decrease in DHT levels in the spinal cord that was reverted by DHT or 3α-diol treatments. In addition, 3α-diol treatment resulted in a significant increase in 3α-diol in the spinal cord compared with control values. Both steroids showed analgesic properties on diabetic neuropathic pain, affecting different pain parameters and possibly by different mechanisms of action. Indeed, DHT counteracted the effect of diabetes on the mechanical nociceptive threshold, pre- and post-synaptic components, glutamate release, astrocyte immunoreactivity, and expression of interleukin-1β (IL1β), while 3α-diol was effective on tactile allodynia threshold, glutamate release, astrocyte immunoreactivity and the expression of substance P, toll-like receptor 4, tumor necrosis factor-α, transforming growth factor β-1, IL1β, and translocator protein. These results indicate that testosterone metabolites are potential agents for the treatment of diabetic neuropathic pain.


Contrast transfer function of de-noising algorithms.
This paper presents a comprehensive study on the contrast transfer function of de-noising algorithms. In order to cover a broad variety of methods, 45 de-noising algorithms are chosen considering their recognized efficiency in the different application domains of image processing. Advanced methods are targeted: wavelet transform-based algorithms with Daubechies, symlets, curvelets, contourlets, patch-based methods such as BM3D, NL-means algorithms and deep learning approaches; in addition, classical spatial filtering methods are considered, such as Wiener, median, Gauss filtering, and adaptive filtering approaches such as anisotropic diffusion and synthetic aperture radar filtering. The contrast transfer function is provided for each algorithm. Ranking of the set of de-noising algorithms is established according to proposed metrics. The paper provides practical methodology and novel results dedicated to the evaluation of the contrast transfer function of de-noising approaches from literature.


Exploring the prevalence of and factors associated with pain: a cross-sectional study of community-dwelling people with dementia.
Few pain studies have made community-dwelling people with dementia (PWD) their focus. The aim of this study was to determine the prevalence of pain among this patient population and to explore medication use. Moreover, we sought to investigate patient and caregiver variables associated with the presence of pain. Community-dwelling PWD and their caregivers were recruited between May 2009 and July 2012 from outpatient memory clinics in Northern Ireland to take part in a face-to-face structured interview with a researcher. Patients' cognitive status and presence of depression were established. A full medication history was taken. Both patients and caregivers were asked to rate patients' pain, at the time of the interview and on an average day, using a 7-point verbal descriptor scale. From the 206 patients who were eligible to take part, 75 patient-caregiver dyads participated in the study (participation rate = 36.4%). The majority of patients (92.0%) had dementia classed as mild or moderate. Pain was commonly reported among the sample, with 57.3% of patients and 70.7% of caregivers reporting patient pain on an average day. Significant differences were found between patients' and caregivers' reports of pain. Two-fifths of patients (40.0%) were prescribed analgesia. Antipsychotic, hypnotic and anxiolytic drug use was low, whereas antidepressant drugs were prescribed more commonly. Presence of pain was unaffected by dementia severity; however, the use of prescribed analgesic medication was a significant predictor of the presence of pain in these patients, whether reported by the patient or their caregiver 'right now' or 'on an average day' (P < 0.001). Patient and caregiver recruitment was challenging, and remains a barrier to research in this area in the future.


Effect of Ptosis Surgery on Tear Oxidative Stress Levels in Patients with Blepharoptosis and Pseudoptosis.
Investigation of the surgical correction effect on tear film functions and tear oxidative stress levels in patients with blepharoptosis and pseudoptosis. Sixty patients with blepharoptosis or pseudoptosis due to dermatochalasis and 30 healthy controls were included in the study. Forty eyes underwent upper blepharoplasty and 20 eyes underwent levator surgery. The tear oxidative stress levels (8-hydroxydeoxyguanosine [8-OHdG] and 4-hydroxy-2-nonenal [4-HNE]) were determined by enzyme-linked immunosorbent assay and tear film functions were evaluated pre-operatively and at the post-operative 1st and 6th months. 8-OHdG and 4-HNE levels in tears were found higher in patients with dermatochalasis (86.3±38.2 ng/mL; 29.8±11.4 ng/mL, respectively) and blepharoptosis (95.3±43.8 ng/mL; 40.8±3.8 ng/mL, respectively) compared to healthy controls (52.9±14.0 ng/mL; 27.8±6.6 ng/mL, respectively). Both levels decreased 1 month after blepharoplasty surgery. The 8-OHdG level in tears of patients who underwent levator surgery increased 1 month after the surgery (p=0.008). No change was detected in tear function tests findings between visits in any patient group. Dermatochalasis and blepharoptosis may lead to an increase in the tear oxidative stress levels. Contrary to a decrease in these levels after blepharoplasty, they may increase in the early period after levator surgery followed by a return to normal levels at the 6th-month visit.


[Ulceromutilating acroosteopathy of the feet (author's transl)].
The cutaneo-osseous syndrome of ulceromutilating acroosteopathy, occurring in a rare hereditary and a "symptomatic" form, represents a nosologically clearly defined disease of the forefoot. Main symptoms are painless, deeply penetrating plantar ulcers with hyperkeratotic edges, non-segmental peripheral sensory neuropathy with a more or less dissociated disturbance of sensitivity and pyrexia of the forefoot (or single toes) without typical clinical signs of inflammation. Secondary symptoms, obligatory in therapy-resistant cases, include (almost) non-reactive acroosteolyses of the "neuropathic" type. The disease, increasing potentially in frequency and not recognised sufficiently, can only be treated successfully when pathogenic risk and precipitation factors are abolished selectively. Twenty-five cases could be observed over a period of 11 years with an increasing incidence.


[Morphological alterations of the skeletal muscles in mitochondrial myopathies in children and their mothers].
Skeletal muscle biopsies from 18 children and 8 from their mothers were studied in mitochondrial myopathies. It is shown that by means of electron microscopy, histochemistry, and morphometry the correct diagnosis of mitochondrial insufficiency is quite possible.


Using a correction factor to correct for overreporting in a food-frequency questionnaire does not improve biomarker-assessed validity of estimates for fruit and vegetable consumption.
To correct for overreporting of fruit and vegetable (FV) consumption in a food-frequency questionnaire, summary questions about consumption of main FV groups are often used to calculate correction factors. This study compared the ability to rank people according to their FV intake of those summary questions and the sum of questions on individual FV items within categories, and of corrected or uncorrected estimates of specific sorts of FV. Healthy middle-age women (n = 161) completed a food-frequency questionnaire about FV consumption during the previous month and gave a single fasting blood sample. Correction factors were calculated as the reported frequency on a summary question divided by the summed frequencies of all items in a category. Plasma carotenoids and vitamin C served as biomarkers of FV consumption. Significant correlations between FV consumption and biomarkers were observed (e.g., Spearman's correlation coefficient r with total carotenoids/vitamin C: 0.32/0.34 for vegetables, 0.30/0.25 for fruits). Summary estimates of cooked, raw and total vegetable consumption correlated higher with biomarkers than sum estimates. For fruits no differences in correlations between sum and summary estimates were observed. Applying a correction factor on the consumption of carrots and total cabbage resulted in lower correlations with relevant biomarkers. For broccoli/cauliflower, Brussels sprouts and citrus fruits, correlations with biomarkers did not change after correction. We conclude that summary questions may suffice to rank individuals according to their intake of main FV categories, and that correction for overreporting of individual FV items is probably not advisable when ranking individuals according to intake of these items.


Lamotrigine in rapid-cycling bipolar disorder.
We evaluated the antidepressant and mood-stabilizing effects of lamotrigine, a novel anticonvulsant, in a group of rapid-cycling bipolar patients. Most were already nonresponders or poor partial responders to other conventional mood-stabilizing agents. This open, naturalistic, and prospective study was conducted with five rapid-cycling bipolar patients (DSM-IV). Each received lamotrigine titrated to a minimum dose of 150 mg/day as monotherapy or in combination with other psychotropic agents. Patients were assessed with the Global Assessment Scale (GAS), Beck Depression Inventory (BDI), and Young Mania Rating Scale (YMRS) for evidence of cycling mood. Lamotrigine was used at a mean +/- SD dose of 185.0 +/- 33.5 mg/day for 225.8 +/- 28.0 days. Random regression modeling of data showed significant dose- and time-dependent improvements in depressive symptoms and social function of patients taking lamotrigine (Dose: z = 2.17, p < .03 for BDI, z = 4.44, p < .001 for GAS; Time: z = -3.79, p < .001 for BDI, z = 2.16, p < .03 for GAS). Further random regression modeling analysis of change over time in symptoms prior to lamotrigine compared with symptoms during lamotrigine treatment showed a significant treatment by time effect for GAS (z = 2.40, p < .016) and a trend for BDI scores (z = -1.79, p < .073). No significant time or dosage effect or time by treatment effect was observed for YMRS. Finally, t statistics showed a significant reduction in mean BDI scores following treatment with lamotrigine (t = -5.26, p < .006). Lamotrigine was well tolerated by all patients; only one patient experienced several side effects, which were probably due to interaction between several psychotropic medications. Lamotrigine augmentation therapy and monotherapy appeared to have mood-stabilizing and antidepressant efficacy in the treatment of five rapid-cycling bipolar patients. The effect persisted for an average of 7.5 months.


Prediction of orientation relationships and interface structures between α-, β-, γ-FeSi2 and Si phases.
A pure crystallogeometrical approach is proposed for predicting orientation relationships, habit planes and atomic structures of the interfaces between phases, which is applicable to systems of low-symmetry phases and epitaxial thin film growth. The suggested models are verified with the example of epitaxial growth of α-, γ- and β-FeSi2 silicide thin films on silicon substrates. The density of near-coincidence sites is shown to have a decisive role in the determination of epitaxial thin film orientation and explains the superior quality of β-FeSi2 thin grown on Si(111) over Si(001) substrates despite larger lattice misfits. Ideal conjunctions for interfaces between the silicide phases are predicted and this allows for utilization of a thin buffer α-FeSi2 layer for oriented growth of β-FeSi2 nanostructures on Si(001). The thermal expansion coefficients are obtained within quasi-harmonic approximation from the DFT calculations to study the influence of temperature on the lattice strains in the derived interfaces. Faster decrease of misfits at the α-FeSi2(001)||Si(001) interface compared to γ-FeSi2(001)||Si(001) elucidates the origins of temperature-driven change of the phase growing on silicon substrates. The proposed approach guides from bulk phase unit cells to the construction of the interface atomic structures and appears to be a powerful tool for the prediction of interfaces between arbitrary phases for subsequent theoretical investigation and epitaxial film synthesis.


Minoxidil sulfotransferase enzymatical activity in plants: A novel paradigm in increasing minoxidil response in androgenetic alopecia.
Minoxidil is the only US FDA approved topical drug for the treatment of androgenetic alopecia (AGA). Minoxidil is effective in hair re-growth in 30%-40% of patients and 50% of males. To exert its hair growing effect, minoxidil must be sulfonated in the scalp by the minoxidil sulfotransferase enzyme (SULT1A1). Low scalp SULT1A1 correlates with lack of minoxidil response; thus, supplementing the scalp SULT1A1 with naturally occurring minoxidil sulfotransferase enzymes could potentially improve treatment outcomes in AGA patients. In this study, we set to characterize SULT1A1 activity in various plants. From the 10 common botanical extracts we have studied, seven exhibited significant activity toward minoxidil as a substrate; thus, providing a potential novel paradigm to increase minoxidil response with natural supplements. To the best of our knowledge, this is the first study to characterize naturally occurring minoxidil sulfotransferase enzymes in plants.


Leprosy: a review of laboratory and therapeutic aspects--part 2.
Leprosy is a chronic infectious condition caused by Mycobacterium leprae(M. leprae). It is endemic in many regions of the world and a public health problem in Brazil. Additionally, it presents a wide spectrum of clinical manifestations, which are dependent on the interaction between M. leprae and host, and are related to the degree of immunity to the bacillus. The diagnosis of this disease is a clinical one. However, in some situations laboratory exams are necessary to confirm the diagnosis of leprosy or classify its clinical form. This article aims to update dermatologists on leprosy, through a review of complementary laboratory techniques that can be employed for the diagnosis of leprosy, including Mitsuda intradermal reaction, skin smear microscopy, histopathology, serology, immunohistochemistry, polymerase chain reaction, imaging tests, electromyography, and blood tests. It also aims to explain standard multidrug therapy regimens, the treatment of reactions and resistant cases, immunotherapy with bacillus Calmette-Guérin (BCG) vaccine and chemoprophylaxis.


Validation of the English version of the Mood Rhythm Instrument.
Disruption of biological rhythms has been linked to the pathophysiology of mental disorders. However, little is known regarding the rhythmicity of mood symptoms due to the lack of validated clinical questionnaires. A better understanding of the rhythmicity of mood symptoms can help identifying individuals whose severity of mood symptoms follows an altered circadian rhythm. The objective of this study was to validate the English version of the Mood Rhythm Instrument (MRhI), a self-reported measure of self-perceived rhythmicity of mood symptoms and behaviours, in a sample of the general population from Canada. After the translation process, the final English version of the Mood Rhythm Instrument (MRhI-English) was applied on participants recruited at McMaster University and St. Joseph's Healthcare Hamilton campuses. Individuals were also asked to answer the Reduced Morningness-Eveningness Questionnaire (rMEQ). Four hundred one individuals completed the English version of the MRhI and the rMEQ. The MRhI-English presented a Cronbach's alpha of 0.75. The factorial analysis grouped the MRhI-15 items in 3 factors (cognitive, affective and somatic), with affective items having a lower frequency of self-reported 24-h peaks. Comparison between sexes showed that women reported a higher frequency of daily peaks in irritability, anxiety, sadness and talking to friends, while men exhibited peaks more frequently in problem-solving, sexual arousal and motivation to exercise. Our findings suggest that the English version of the MRhI displayed good internal consistency. Future directions will include the use of the MRhI instrument in individuals with mood disorders, aiming to provide a better understanding of the relationship between daily patterns of mood variability and mental health outcomes.


Synthesis, stabilities, and redox behavior of Di(1-azulenyl)(6-azulenyl)methylium hexafluorophosphates. Generation of a donor-acceptor-substituted neutral radical by azulenes.
Several di(1-azulenyl)(6-azulenyl)methanes and 1,3-bis[(1-azulenyl)(6-azulenyl)methyl]azulenes were prepared by the condensation reaction of azulenes with diethyl 6-formylazulene-1,3-dicarboxylate under acidic conditions. The products were converted into di(1-azulenyl)(6-azulenyl)methylium hexafluorophosphates and azulene-1,3-diylbis[(1-azulenyl)(6-azulenyl)methylium] bis(hexafluorophosphate)s via hydride abstraction reaction with DDQ following the exchange of counterions. These mono- and dications exhibited high stability with large pK(R)(+) values (5.6-10.1), despite the captodative substitution of azulenes. The electrochemical reduction of the monocations upon cyclic voltammetry (CV) exhibited a reversible two-step, one-electron reduction wave with a small difference between the first reduction potential (E(1)(red)) and the second one (E(2)(red)), which exhibited the generation of highly amphoteric neutral radicals in solution. The electrochemical reduction of dications showed voltammograms, which were characterized by subsequent two single-electron waves and a two-electron transfer upon CV attributable to the formation of a radical cation, a diradical (or twitter ionic structure), and a dianionic species, respectively. Formation of a persistent neutral radical from a monocation was revealed by ESR and UV-vis spectroscopies and theoretical calculations. The ESR spectra of the neutral radical gave two hyperfine coupling constants: a(H) = 0.083 (6H) and 0.166 mT (9H) (g = 2.0024), indicating that an unpaired electron delocalizes over all three of the azulene rings. The stable monoanion, which shows the localization of the charge on the 6-azulenyl substituent, was also successfully generated from the di(1-azulenyl)(6-azulenyl)methane derivative.


Translational regulation of the JunD messenger RNA.
JunD, a member of the Jun family of nuclear transcription proteins, dimerizes with Fos family members or other Jun proteins (c-Jun or JunB) to form the activator protein 1 (AP-1) transcription factor. The junD gene contains no introns and generates a single mRNA. Here we show that two predominant JunD isoforms are generated by alternative initiation of translation, a 39-kDa full-length JunD protein (JunD-FL) by initiation at the first AUG codon downstream of the mRNA 5' cap and a shorter, 34-kDa JunD protein (DeltaJunD) by initiation at a second in-frame AUG codon. The JunD mRNA contains a long, G/C-rich 5'-untranslated region that is predicted to be highly structured and is important for regulating the ratio of JunD-FL and DeltaJunD protein expression. A third functional out-of-frame AUG directs translation from a short open reading frame positioned between the JunD-FL and DeltaJunD start sites. In addition, three non-AUG codons also support translation, an ACG codon (in-frame with JunD) and a CUG are positioned in the 5'-untranslated region, and a CUG codon (also in-frame with JunD) is located downstream of the short open reading frame. Mutation of these start sites individually had no affect on DeltaJunD protein levels, but mutation of multiple upstream start sites led to an increase in DeltaJunD protein levels, indicating that these codons can function cumulatively to suppress DeltaJunD translation. Finally, we show that the JunD mRNA does not possess an internal ribosome entry site and is translated in a cap-dependent manner.


Effect of glucose on the growth hormone response to L-dopa in normal and diabetic subjects.
The effect of hyperglycemia on the growth hormone response to oral L-dopa (500 mg.) was assessed in eight normal and eight insulin-dependent diabetic subjects. A peak growth hormone response of 21.0 +/- 4.0 ng./ml. (mean +/- S.E.M.), significantly above baseline (p less than 0.01), was achieved in the normal group following oral L-dopa. Glucose concentrations did not change and were approximately 80 mg./100 ml. throughout. Administration of 100 gm. oral glucose with the L-dopa, or thirty minutes thereafter, totally suppressed the growth hormone response in all eight and six of the subjects, respectively. A peak growth hormone response of 20.0 "/- 1.7 ng./ml. (mean +/- S.E.M.), significantly above baseline (p less than 0.001), was obtained in eight nonobese, insulin-dependent diabetics, in spite of prevailing hyperglycemia (mean plasma glucose 243-258 mg./100 ml.) throughout the test. Endogenous hyperglycemia was achieved in these patients by lessening the usual strict adherence to plasma glucose control for the purpose of the study. These results suggest an abnormality in the hypothalamus or pituitary of diabetic subjects allowing growth hormone responsiveness in spite of hyperglycemia.


Phellodendron amurense bark extract prevents progression of prostate tumors in transgenic adenocarcinoma of mouse prostate: potential for prostate cancer management.
Prostate cancer is the second leading cause of cancer-related deaths in men in Western society. Epidemiological studies suggest that a reduced risk of cancer is associated with the consumption of a phytochemical-rich diet that includes fruits and vegetables. Strategies to delay clinically significant prostate cancer will have a tremendous impact in reducing the overall incidence of prostate cancer as well as improving quality of life for elderly men. Furthermore, the long latency involved in the development of clinically significant prostate cancer provides a plethora of opportunities for its management, especially using prevention approaches. Previous studies from our laboratory show that Nexrutine (bark extract from Phellodendron amurense) prevents prostate tumor development when given prior to the development of high-grade prostatic intraepithelial neoplasia in the transgenic adenocarcinoma of mouse prostate (TRAMP) model. In this study, we investigated the effect on the progression of established tumors in the TRAMP model by administering Nexrutine to 28-week-old TRAMP mice. Efficacy of Nexrutine was determined by histopathological evaluation of the prostate. Our data indicate that Nexrutine inhibited progression of prostate tumors that was correlated with tissue levels of transcription factors nuclear factor kappa B, cyclic-AMP response element-binding protein and phosphorylated CREB. Moreover, Nexrutine intervention resulted in a significant increase in the bone mineral density of the left femur diaphysis (p=0.009) and prevented the development of metastatic lesions. Nexrutine treatment also significantly (p=0.005) inhibited invasion of androgen-independent prostate cancer cells.


CRISPR-Cas9-assisted recombineering in Lactobacillus reuteri.
Clustered regularly interspaced palindromic repeats (CRISPRs) and the CRISPR-associated (Cas) nuclease protect bacteria and archeae from foreign DNA by site-specific cleavage of incoming DNA. Type-II CRISPR-Cas systems, such as the Streptococcus pyogenes CRISPR-Cas9 system, can be adapted such that Cas9 can be guided to a user-defined site in the chromosome to introduce double-stranded breaks. Here we have developed and optimized CRISPR-Cas9 function in the lactic acid bacterium Lactobacillus reuteri ATCC PTA 6475. We established proof-of-concept showing that CRISPR-Cas9 selection combined with single-stranded DNA (ssDNA) recombineering is a realistic approach to identify at high efficiencies edited cells in a lactic acid bacterium. We show for three independent targets that subtle changes in the bacterial genome can be recovered at efficiencies ranging from 90 to 100%. By combining CRISPR-Cas9 and recombineering, we successfully applied codon saturation mutagenesis in the L. reuteri chromosome. Also, CRISPR-Cas9 selection is critical to identify low-efficiency events such as oligonucleotide-mediated chromosome deletions. This also means that CRISPR-Cas9 selection will allow identification of recombinant cells in bacteria with low recombineering efficiencies, eliminating the need for ssDNA recombineering optimization procedures. We envision that CRISPR-Cas genome editing has the potential to change the landscape of genome editing in lactic acid bacteria, and other Gram-positive bacteria.


New hybrid films based on cellulose and hydroxygallium phthalocyanine. Synergetic effects in the structure and properties.
Hydroxygallium phthalocyanine (HOGaPc) and cellulose (from a trimethylsilyl derivative) have been used as native elements for the preparation of a novel family of hybrid films. By spin-coating, both components allow the building of films with different configurations on various substrates in a controlled way. The particularities of these hybrid films have been characterized by a range of techniques such as Fourier transform infrared spectroscopy (FTIRS) in attenuated total reflection using multiple internal reflections (ATR/MIR), absorption ultraviolet and visible spectroscopy (UV-vis), X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), and surface potential measurements using the Kelvin-Zisman vibrating capacitor probe (KP). This enabled determination of the influence of cellulose on the arrangement of HOGaPc and, consequently, control of the relation between the structure and the properties of the films. Finally, gas sensor tests were performed to check the potentialities of these hybrid films. In particular, the synergetic behavior between the film-forming materials allows a fast and sensible change in surface potential after cyclic exposures to ozone (O3, 100 ppb) and nitrogen. Overall, we present the advantages of combining phthalocyanine with cellulose in enhancing the properties of the final product. Introduction of cellulose as a host material opens up a new area of hybrid films.


Public health surveillance of automated external defibrillators in the USA: protocol for the dynamic automated external defibrillator registry study.
Lay use of automated external defibrillators (AEDs) before the arrival of emergency medical services (EMS) providers on scene increases survival after out-of-hospital cardiac arrest (OHCA). AEDs have been placed in public locations may be not ready for use when needed. We describe a protocol for AED surveillance that tracks these devices through time and space to improve public health, and survival as well as facilitate research. Included AEDs are installed in public locations for use by laypersons to treat patients with OHCA before the arrival of EMS providers on scene. Included cases of OHCA are patients evaluated by organised EMS personnel and treated for OHCA. Enrolment of 10 000 AEDs annually will yield precision of 0.4% in the estimate of readiness for use. Enrolment of 2500 patients annually will yield precision of 1.9% in the estimate of survival to hospital discharge. Recruitment began on 21 Mar 2014 and is ongoing. AEDs are found by using multiple methods. Each AED is then tagged with a label which is a unique two-dimensional (2D) matrix code; the 2D matrix code is recorded and the location and status of the AED tracked using a smartphone; these elements are automatically passed via the internet to a secure and confidential database in real time. Whenever the 2D matrix code is rescanned for any non-clinical or clinical use of an AED, the user is queried to answer a finite set of questions about the device status. The primary outcome of any clinical use of an AED is survival to hospital discharge. Results are summarised descriptively. These activities are conducted under a grant of authority for public health surveillance from the Food and Drug Administration. Results are provided periodically to participating sites and sponsors to improve public health and quality of care.


Venom yield and its relationship with body size and fang separation of pit vipers from Argentina.
The amount of venom that a snake can inject is related to its body size. The body size is related to head size and to the distance between fangs. To correlate snake body size, distance between fangs and distance between puncture wounds with the venom yield (and consequently with the venom dose potentially injected in a single snakebite), we studied these variables in two species of public health importance in South America, Bothrops (Rhinocerophis) alternatus, and Crotalus durissus terrificus. In all cases a positive correlation was observed between body length, fang separation distance, distance between puncture wounds and venom yield, with a regression coefficient over 0.5 for Bothrops alternatus and over 0.6 for Crotalus durissus terrificus in all cases, being the relation distance between punctures wounds and venom yield of 0.54 and 0.69 respectively. The difference between fang separation and puncture separation was never greater than 30%, with a mean difference around 13%. The strong relationships between body size, fang separation and venom yield may be useful for planning potential venom production in serpentariums. In addition, because puncture mark separation gives an approximate idea of the size of the snake, this provides a rough idea of the size of the snake that produced a bite and the potential amount of venom that could have been injected.


Detrimental effects of antiapoptotic treatments in spinal cord injury.
Long-term functional impairments due to spinal cord injury (SCI) in the rat result from secondary apoptotic death regulated, in part, by SCI-induced decreases in protein levels of the antiapoptotic protein Bcl-xL. We have shown that exogenous administration of Bcl-xL spares neurons 24 h after SCI. However, long-term effects of chronic application of Bcl-xL have not been characterized. To counteract SCI-induced decreases in Bcl-xL and resulting apoptosis, we used the TAT protein transduction domain fused to the Bcl-xL protein (Tat-Bcl-xL), or its antiapoptotic domain BH4 (Tat-BH4). We used intrathecal delivery of Tat-Bcl-xL, or Tat-BH4, into injured spinal cords for 24 h or 7 days, and apoptosis, neuronal death and locomotor recovery were assessed up to 2 months after injury. Both, Tat-Bcl-xL and Tat-BH4, significantly decreased SCI-induced apoptosis in thoracic segments containing the site of injury (T10) at 24 h or 7 days after SCI. However, the 7-day delivery of Tat-Bcl-xL, or Tat-BH4, also induced a significant impairment of locomotor recovery that lasted beyond the drug delivery time. We found that the 7-day administration of Tat-Bcl-xL, or Tat-BH4, significantly increased non-apoptotic neuronal loss and robustly augmented microglia/macrophage activation. These results indicate that the antiapoptotic treatment targeting Bcl-xL shifts neuronal apoptosis to necrosis, increases the inflammatory response and impairs locomotor recovery. Our results suggest that a combinatorial treatment consisting of antiapoptotic and anti-inflammatory agents may be necessary to achieve tissue preservation and significant improvement in functional recovery after SCI.


[Oral discoid lupus erythematosus. Diagnostic considerations apropos of a case].
The study and presentation of a typical case of discoid lupus erythematosus with oral lesions in a 30-years old woman, without visceral manifestations at present, was the motive for a revision and updating of the concepts of etiology, differential diagnosis, treatment and prognosis of this disease. The authors emphasize the importance of the clinical and evolving aspect of the lesions, long-term development, as well as the response to treatment (an antimalarial synthesis, applied systematically and infiltrates with betamethasone solution) in order to reach valid diagnostic conclusions. It is important in the anatomopathological study to include the atrophic zones of the central portion of the discoid lesion.


Natural Compounds As Modulators of Non-apoptotic Cell Death in Cancer Cells.
Cell death is an innate capability of cells to be removed from microenvironment, if and when they are damaged by multiple stresses. Cell death is often regulated by multiple molecular pathways and mechanism, including apoptosis, autophagy, and necroptosis. The molecular network underlying these processes is often intertwined and one pathway can dynamically shift to another one acquiring certain protein components, in particular upon treatment with various drugs. The strategy to treat human cancer ultimately relies on the ability of anticancer therapeutics to induce tumor-specific cell death, while leaving normal adjacent cells undamaged. However, tumor cells often develop the resistance to the drug-induced cell death, thus representing a great challenge for the anticancer approaches. Numerous compounds originated from the natural sources and biopharmaceutical industries are applied today in clinics showing advantageous results. However, some exhibit serious toxic side effects. Thus, novel effective therapeutic approaches in treating cancers are continued to be developed. Natural compounds with anticancer activity have gained a great interest among researchers and clinicians alike since they have shown more favorable safety and efficacy then the synthetic marketed drugs. Numerous studies in vitro and in vivo have found that several natural compounds display promising anticancer potentials. This review underlines certain information regarding the role of natural compounds from plants, microorganisms and sea life forms, which are able to induce non-apoptotic cell death in tumor cells, namely autophagy and necroptosis.


Vasospastic Angina Presenting With Syncope and Chest Pain: A Case Report and Brief Literature Review.
A 65-year-old male presented to the hospital with chest pain associated with recurrent syncope. He had a history of coronary artery disease and a long-standing history of smoking. While he was hospitalized, he had an episode of chest pain during which he was found to have transient ST segment elevation in the inferior leads. He was also noted to have a brief cardiac tachyarrhythmia. Coronary arteriography revealed vasospasm of the left anterior descending artery and right coronary artery, which were relieved to a significant extent after administration of intracoronary nitroglycerin. Subsequent angiograms and fractional flow reserve studies, demonstrated underlying non-obstructive coronary artery disease at the sites of spasm. No percutaneous coronary intervention was pursued. The patient was started on a calcium channel blocker on dismissal from the hospital. Upon follow up several months later, he remained free of symptoms that brought him to the hospital.


The effectiveness and safety of TNF-alpha blockers in the treatment of early psoriatic arthritis: an Italian multicentre longitudinal observational pilot study.
The objective of this study is to assess the effectiveness and safety of TNF-α blockers in a group of early psoriatic arthritis (PsA) patients with an unsatisfactory response to previous conventional treatment consecutively enrolled in five Italian centres. A 24-week open-label trial was carried out in consecutive early PsA patients classified according to the CASPAR criteria, with unsatisfactory response to previous treatments and with a DAS28 threshold as ≥3.2, seen at the outpatient clinics of each centre. Exclusion criteria were previous usage of TNF-α blockers and a disease duration >12 months. The choice of any of the three TNF-α blockers was decided by the expert's opinion, without any restriction. Effectiveness was considered as an improvement of DAS28 at 12 and 24 weeks of treatment. Secondary endpoints were an improvement of TJC, SWJ, HAQ score and PASI score. Changes from baseline to the 12- and 24-week follow-up assessments were analysed using the Wilcoxon paired sign rank test. Twenty-nine patients (14 males, 15 females, median age 37 years, range 20-65 years) were enrolled. A statistical improvement of the DAS28 was observed at 12 and 24 weeks from baseline (p<0.001). Secondary endpoints also confirmed the effectiveness of the TNF-α blockers in the treatment of early PsA. No severe adverse events were observed during the treatment period, and no patient withdrew from the medications. This study suggests that the TNF-α blockers can be effective in the management of early PsA. Further controlled studies will provide more data on this challenging topic.


Magnetostructural dynamics of Rieske versus ferredoxin iron-sulfur cofactors.
The local chemical environment of the [2Fe-2S] cofactor hosted by ferredoxin and Rieske-type proteins is fundamentally different due to the presence of distinct ligands at the two iron centers in the case of Rieske proteins, whereas they are identical in ferredoxins. This renders Rieske [2Fe-2S] cores chemically asymmetric and results in more complex vibrational spectra as compared to ferredoxin. Likewise, one would expect other properties, for instance the dynamics of the magnetic exchange coupling constant J, to be also more complex. Applying ab initio molecular dynamics using our recently introduced spin-constrained two-determinant extended broken symmetry (CEBS) approach to Rieske and ferredoxin model complexes at 300 K, we extract the molecular fluctuations and the resulting magnetostructural cross-correlations involving the antiferromagnetic exchange interaction J(t). This analysis demonstrates that the details of the magnetostructural dynamics are indeed distinctly different for Rieske and ferredoxin cofactors, while the time averages of 〈J〉 are shown to be essentially identical. In particular, the frequency window between about 200 and 350 cm(-1), is a "fingerprint region" that allows one to distinguish chemically asymmetric from symmetric cofactors and thus Rieske proteins from ferredoxins.


New perspective on parkinsonism in frontotemporal lobar degeneration.
Frontotemporal dementia (FTD) is the second most common type of presenile dementia. Three clinical prototypes have been defined; behavioral variant FTD, semantic dementia, and progressive nonfluent aphasia. Progressive supranuclear palsy, corticobasal degeneration, and motor neuron disease may possess clinical and pathological characteristics that overlap with FTD, and it is possible that they may all belong to the same clinicopathological spectrum. Frontotemporal lobar degeneration (FTLD) is a clinicopathological syndrome that encompasses a heterogenous group of neurodegenerative disorders. Owing to the advancement in the field of molecular genetics, diagnostic imaging, and pathology, FTLD has been the focus of great interest. Nevertheless, parkinsonism in FTLD has received relatively less attention. Parkinsonism is found in approximately 20-30% of patients in FTLD. Furthermore, parkinsonism can be seen in all FTLD subtypes, and some patients with familial and sporadic FTLD can present with prominent parkinsonism. Therefore, there is a need to understand parkinsonism in FTLD in order to obtain a better understanding of the disease. With regard to the clinical characteristics, the akinetic rigid type of parkinsonism has predominantly been described. Parkinsonism is frequently observed in familial FTD, more specifically, in FTD with parkinsonism linked to chromosome 17q (FTDP-17). The genes associated with parkinsonism are microtubule associated protein tau (MAPT), progranulin (GRN or PGRN), and chromosome 9 open reading frame 72 (C9ORF72) repeat expansion. The neural substrate of parkinsonism remains to be unveiled. Dopamine transporter (DAT) imaging revealed decreased uptake of DAT, and imaging findings indicated atrophic changes of the basal ganglia. Parkinsonism can be an important feature in FTLD and, therefore, increased attention is needed on the subject.


Referral parameters for diagnosis of glaucoma in general eye clinic at Tilganga Institute of Ophthalmology.
To find out the most common referral parameter among the glaucoma suspects patients from general eye clinic and to establish glaucoma diagnosis. This study is a retrospective cohort hospital based study. Two hundred patients from January to February 2017 sent to glaucoma clinic as glaucoma suspects were re-evaluated meticulously by glaucoma specialist and were diagnosed as glaucoma, non glaucoma, suspects and ocular hypertension. Out of the 200 patients referred to glaucoma clinic as glaucoma suspects only19% were diagnosed to have glaucoma. The mean age at which glaucoma diagnosed was 55.29(14.4) compared to 41.6(15.1) in normal group. One hundred and sixty five patients were referred on the basis of suspicious optic nerve head, among them 14.5% (24/165) had glaucoma. This study showed that, open angle glaucoma (OAG) 28.9% was the most common type of total glaucoma diagnosed. The mean vertical cup discratio in the OAG group was 0.69±0.1 (0.4 -0.9) compared to 0.56 ± 0.11((0.2-0.8)(p=0.00) normal. The mean intra ocular pressure (IOP) in OAG group was 19.73 ±4.95(11-32) mmHg compared to 16.74± 3.36(10-30) mmHg (p=0.00) in normal group. The mean central corneal thickness (CCT) in OAG group was 533.05 ± 31.24μm (467-606) compared to normal was 534.9±33.6 μm (432-696) (p=0.670). Suspicious optic nerve head is the most common referral parameter between the general ophthalmologist and residents, but this study shows only few of them were diagnosed with glaucoma. This gives us a clue that the ophthalmologists and residents are to be trained better to help them identify the signs of glaucoma on the optic nerve head beside its size, which will reduce unnecessary burden to the resources of patients and hospital.


Children and natural disasters.
The number of children affected by natural disasters each year is alarmingly high and can be expected to rise as climate change continues. The mental consequences have been documented not only in the rates of post-traumatic stress symptoms and post-traumatic stress disorder, but also for depression and other mental health problems. To contribute towards the knowledge in this area, this special issue of the European Journal of Psychotraumatology focuses on how children can be prepared for natural disasters and the mental health aspects of such events. It includes articles on communicating risks to children, involving children in disaster risk reduction, and the mental health consequences for children from an earthquake, a volcanic eruption, a flood and a hurricane. In this special issue specifically focusing on children and natural disasters, we hope to enhance our understanding of some of the complex pathways and pave the way for improving our interventions.


A comparison of postoperative, three-dimensional soft tissue changes in patients with skeletal class III malocclusions treated via orthodontics-first and surgery-first approaches.
The aim of this retrospective study was to compare three-dimensional (3D) soft tissue and hard tissue changes between orthodontics-first approach (OFA) and surgery-first approach (SFA) after mandibular setback surgery. All patients underwent bilateral sagittal split osteotomy, and were examined by lateral cephalograms and 3D optical scanner before surgery (T0) and 1 (T1), 3 (T2), and 12 (T3) months after surgery. Three standard angles (FMA, U1 to FH, IMPA) were measured as hard tissue change and the 2 sets of 3D data were superimposed, and volumetric differences were calculated as soft tissue change. Statistical analyses were performed by using unpaired t-tests. Differences with P < 0.05 were considered significant. A total of 39 patients with mandibular prognathism were included in this study. The OFA group consisted of 24 patients and the SFA group of 15 patients. The SFA group exhibited more labial inclination from T1 to T2 (p = 0.008) and T2 to T3 (p = 0.003) than did the OFA group. There were no significant changes at maxilla and mandible at each term of T0, T1, T2 and T3 (p > 0.05), but compared to before surgery, mandibular volume in SFA group significant increased at 1year (p = 0.049) after surgery. We found that the soft tissue changes after the SFA differed significantly from those after the OFA; thus, soft tissue predictions require more care. An analysis of our data compared with OFA and SFA for the patient with mandibular prognathism confirm that the mandibular soft tissue changes by postoperative orthodontic treatment and occlusal relationship in SFA.


Agomelatine: a novel antidepressant.
Depression is highly prevalent in community and primary care samples. According to the findings of the Sequential Treatment Alternatives to Relieve Depression study, after multiple clinical interventions, approximately one-third of patients never fully experienced remission from depression. This somber finding invites consideration of novel antidepressant options, which may in the near future include agomelatine. Agomelatine is a melatonergic agonist and a 5HT(2c) antagonist (i.e., it has a unique mechanism of action). The melatonergic function appears to improve sleep patterns, whereas the serotonergic antagonism results in the release of norepinephrine and dopamine. Given the current information, the overall side-effect profile of agomelatine appears relatively mild. For example, agomelatine has no discontinuation syndrome, exhibits infrequent sexual dysfunction, and is generally weight neutral. The drug appears to be relatively safe in overdose. On a cautionary note, however, one percent of patients experience elevated hepatic transaminases while on treatment. Overall, agomelatine may be a unique pharmacological addition in the clinical war on depression in both psychiatric and primary care settings-if further evaluation in clinical trials supports reasonable risk.


Intracellular distribution of Photofrin in malignant and normal endothelial cell lines.
Compared to current treatments including surgery, radiation therapy, and chemotherapy, PDT offers the advantage of an effective and selective method of destroying diseased tissues without damaging surrounding healthy tissues. One of the aspects of antitumour effectiveness of PDT is related to the distribution of photosensitizing drugs. The localization of photosensitizers in cytoplasmic organelles during PDT plays a major role in the cell destruction; therefore, intracellular localization of Ph in malignant and normal cells was investigated. The cell lines used throughout the study were: human malignant A549, MCF-7, Me45 and normal endothelial cell line HUV-EC-C. After incubation with Ph cells were examined using fluorescence and confocal microscopy to visualize the photosensitizer accumulation. For cytoplasm and mitochondria identification, cells were stained with CellTracker Green and MitoTracker Green, respectively. Distribution of Ph was different in malignant and normal cells and dependent on the incubation time. The maximal concentration of Ph in two malignant cell lines (A549 and MCF-7) was observed after 4 hours of incubation, and the most intensive signal was observed around the nuclear envelope. Intracellular distribution of Ph in the Me45 cell line showed that the fluorescence emitted by Ph overlaid that from MitoTracker. This indicates preferential accumulation of the sensitizer in mitochondria. Our results based on the mitochondrial localization support the idea that PDT can contribute to elimination of malignant cells by inducing apoptosis, which is of physiological significance.


Molecular predictors of sensitivity to the insulin-like growth factor 1 receptor inhibitor Figitumumab (CP-751,871).
Figitumumab (CP-751,871), a potent and fully human monoclonal anti-insulin-like growth factor 1 receptor (IGF1R) antibody, has been investigated in clinical trials of several solid tumors. To identify biomarkers of sensitivity and resistance to figitumumab, its in vitro antiproliferative activity was analyzed in a panel of 93 cancer cell lines by combining in vitro screens with extensive molecular profiling of genomic aberrations. Overall response was bimodal and the majority of cell lines were resistant to figitumumab. Nine of 15 sensitive cell lines were derived from colon cancers. Correlations between genomic characteristics of cancer cell lines with figitumumab antiproliferative activity revealed that components of the IGF pathway, including IRS2 (insulin receptor substrate 2) and IGFBP5 (IGF-binding protein 5), played a pivotal role in determining the sensitivity of tumors to single-agent figitumumab. Tissue-specific differences among the top predictive genes highlight the need for tumor-specific patient selection strategies. For the first time, we report that alteration or expression of the MYB oncogene is associated with sensitivity to IGF1R inhibitors. MYB is dysregulated in hematologic and epithelial tumors, and IGF1R inhibition may represent a novel therapeutic opportunity. Although growth inhibitory activity with single-agent figitumumab was relatively rare, nine combinations comprising figitumumab plus chemotherapeutic agents or other targeted agents exhibited properties of synergy. Inhibitors of the ERBB family were frequently synergistic and potential biomarkers of drug synergy were identified. Several biomarkers of antiproliferative activity of figitumumab both alone and in combination with other therapies may inform the design of clinical trials evaluating IGF1R inhibitors.


Inference of segmented color and texture description by tensor voting.
A robust synthesis method is proposed to automatically infer missing color and texture information from a damaged 2D image by (N)D tensor voting (N > 3). The same approach is generalized to range and 3D data in the presence of occlusion, missing data and noise. Our method translates texture information into an adaptive (N)D tensor, followed by a voting process that infers noniteratively the optimal color values in the (N)D texture space. A two-step method is proposed. First, we perform segmentation based on insufficient geometry, color, and texture information in the input, and extrapolate partitioning boundaries by either 2D or 3D tensor voting to generate a complete segmentation for the input. Missing colors are synthesized using (N)D tensor voting in each segment. Different feature scales in the input are automatically adapted by our tensor scale analysis. Results on a variety of difficult inputs demonstrate the effectiveness of our tensor voting approach.


MPTP Mouse Model of Preclinical and Clinical Parkinson's Disease as an Instrument for Translational Medicine.
Parkinson's disease (PD) is characterized by the appearance of motor symptoms many years after the onset of neurodegeneration, which explains low efficiency of therapy. Therefore, one of the priorities in neurology is to develop an early diagnosis and preventive treatment of PD, based on knowledge of molecular mechanisms of neurodegeneration and neuroplasticity in the nigrostriatal system. However, due to inability to diagnose PD at preclinical stage, research and development must be performed in animal models by comparing the nigrostriatal system in the models of asymptomatic and early symptomatic stages of PD. In this study, we showed that despite the progressive loss of neurons in the substantia nigra at the presymptomatic and symptomatic stage, almost no change was observed in the main functional characteristics of this brain region, including dopamine (DA) uptake and release, dopamine transporter (DAT) and vesicular monoamine transporter 2 (VMAT2) expression, and activity of MAO-A and MAO-B. In the striatum of presymptomatic mice, some parameters (DA release and uptake, MAO-A activity) remained compensatory unchanged or compensatory decreased (MAO-B gene expression and activity), while others-a reduction in DA levels in tissue and extracellular space and in VMAT2 and DAT expression-manifest the functional failure. In symptomatic mice, only a few parameters (spontaneous DA release and uptake, MAO-B gene expression and activity) remained at the same level as at presymptomatic stage, while most parameters (DA level in tissue and extracellular space, DA-stimulated release, VMAT2 and DAT contents), decreased, showing decompensation, which was enhanced by increasing MAO-A activity. Thus, this study provides a comprehensive assessment of the molecular mechanisms of neuroplasticity in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine models of preclinical and clinical stages of PD, which could potentially serve as a powerful tool for translational medicine.


Enzymatic resolution, desymmetrization, and dynamic kinetic asymmetric transformation of 1,3-cycloalkanediols.
An efficient desymmetrization of cis-1,3-cyclohexanediol to (1S,3R)-3-(acetoxy)-1-cyclohexanol ((R,S)-2a) was performed via Candida antarctica lipase B (CALB)-catalyzed transesterification, in high yield (up to 93%) and excellent enantioselectivity (ee's up to >99.5%). (R,R)-Diacetate ((R,R)-3a) was obtained in a DYKAT process at room temperature from (1S,3R)-3-acetoxy-1-cyclohexanol ((R,S)-2a), in a high trans/cis ratio (91:9) and in excellent enantioselectivity of >99%. Metal- and enzyme-catalyzed dynamic transformation of cis/trans-1,3-cyclohexanediol using PS-C gave a high diastereoselectivity for cis-diacetate (cis/trans = 97:3). The (1R,3S)-3-acetoxy-1-cyclohexanol (ent-(R,S)-2a) was obtained from cis-diacetate by CALB-catalyzed hydrolysis in an excellent yield (97%) and selectivity (>99% ee). By deuterium labeling it was shown that intramolecular acyl migration does not occur in the transformation of cis-monoacetate to the cis-diacetate.


Severe hyperkalaemia and ketoacidosis during routine treatment with an insulin pump.
During a feasibility study of the use of insulin pumps to treat diabetes ketoacidosis occurred at a rate of 0.14 episodes/patient/year in the first year but was lower in subsequent years. A case of cardiac arrest secondary to hyperkalaemia during ketoacidosis occurred in a patient treated with a pump. The mean (SD) serum potassium concentration on presentation to hospital with ketoacidosis was significantly higher in patients treated with a pump (5.7 (1.1) mmol(mEq)/l) than those treated with conventional injections of insulin (4.9(0.9) mmol/l; p less than 0.01). The high rate of ketoacidosis and raised serum potassium concentrations during treatment with the pump creates doubt about the use of this treatment as an alternative regimen for large numbers of patients in a busy diabetic clinic.


Beyond the RCT - understanding the relationship between interventions, individuals and outcome - the example of neurological rehabilitation.
Rehabilitation is highly complex, involving multiple processes, outcomes and stakeholders. The way we deliver our services and work with our clients and their families should be informed by research approaches that produce the wide range of knowledge needed. This article aims to explore the degree to which the dominant approach to 'evidence' (the randomised clinical trial or RCT) meets those needs and discuss alternate/additional ways of gaining evidence. A critical review of the literature allowing exploration of problems encountered in rehabilitation RCTs and alterative approaches. We discuss some problematic issues related to using RCTs in rehabilitation research (for example the large number of people excluded from trials, and the small numbers of people with some neurological conditions making RCTs non-viable). Alternative approaches are discussed including clinical practice improvement studies (sometimes called practice-based evidence or PBE), which provide data on patients treated in routine practice; qualitative research, which can provide an understanding of the users of health care services to ensure they are meeting their needs; and metasynthesis, which can be used to summarise several qualitative studies to enhance our understanding of the principles underlying service delivery. Finally, we explore how clinicians and commissioners often use evidence generated by forms of research other than the RCT. The best answers about how to enhance rehabilitation outcomes are likely to come from a combination and integration of the most appropriate methods. In conclusion, we urge for more joined up thinking, for learning from different fields so that we can develop more effective and appropriate health care and rehabilitation.


Becoming a Rare Disease Parent: An Interpretative Phenomenological Analysis of Parent-Caregivers' Postpartum Experiences.
Rare diseases constitute a group of conditions that are individually rare, but in aggregate impact between 3 and 6% of the world population. Many of these conditions present during infancy and involve substantial caregiving responsibilities, often assessed via quantitative measurements. However, few qualitative analyses examine lived experiences of parent-caregivers during the early period of their child's life. The purpose of this study was to examine the meaning that rare disease parent-caregivers apply to the postpartum year using data collected from a semi-structured interview exploring significant experiences over the course of their affected child's life. We utilized an interpretative phenomenological analysis (IPA) approach to analyze 22 interview transcripts from caregivers to children with several inherited metabolic and mitochondrial disorders, as well as an undiagnosed disease. Our analysis yielded three superordinate themes: Reckoning With the Parent-Caregiver Role, Familial Transition, and Adaptation and Adjustment. Subordinate themes expanded upon these concepts and included distinctions between the parent and caregiving identity, communal coping and shifting of family dynamics, as well as meaning applied to child milestones, anticipatory grief, and parental perception of a new normal. Exploration of these themes in relation to existing literature, as well as future research directions for qualitative research on rare disease caregivers, is discussed. Overall, this work contributes to a growing body of literature exploring the parental experience of rare disease across several condition contexts.


Diabetes and the accompanying hyperglycemia impairs cardiomyocyte calcium cycling through increased nuclear O-GlcNAcylation.
Diabetic cardiomyopathy is characterized by impaired cardiac contractility leading to poor myocardial performance. We investigated the role that the hexosamine pathway, and especially altered nuclear O-Glc-NAcylation, plays in the development of diabetic cardiomyopathy. Incubating neonatal rat cardiomyocytes in high glucose (25 mM) resulted in prolonged calcium transients when compared with myocytes incubated in normal glucose (5.5 mM), which is consistent with delayed myocardial relaxation. High glucose-treated myocytes also exhibited reduced sarcoendoplasmic reticulum Ca(2+)-ATPase 2a (SERCA2a) mRNA and protein expression, decreased SERCA2a promoter activity, and increased O-GlcNAcylation of nuclear proteins compared with myocytes treated with normal glucose. Exposure of myocytes to 8 mM glucosamine or an adenovirus expressing O-GlcNAc-transferase (OGT) resulted in prolonged calcium transient decays and significantly reduced SERCA2a protein levels, whereas treatment with an adenovirus encoding O-GlcNAcase (GCA) resulted in improved calcium transients and SERCA2a protein levels in myocytes exposed to high glucose. Effects of elevated glucose or altered O-GlcNAcylation were also observed on essential transcription factors involved in cardiomyocyte function. High glucose-treated myocytes (with or without OGT adenovirus) exhibited increased levels of O-GlcNAcylated specificity protein 1 compared with control myocytes, whereas infecting high glucose-treated myocytes with GCA adenovirus reduced the degree of specificity protein 1 Glc-NAcylation. Treatment of myocytes with 25 mM glucose, 8 mM glucosamine, or OGT adenovirus also significantly reduced levels of myocytes enhancer factor-2A protein compared with control myocytes, whereas infection with GCA adenovirus resulted in improved myocytes enhancer factor-2 expression. Our results suggest that the hexosamine pathway, and O-GlcNAcylation in particular, is important in impaired cardiac myocyte function and the development of diabetic cardiomyopathy.


LEU3 of Saccharomyces cerevisiae activates multiple genes for branched-chain amino acid biosynthesis by binding to a common decanucleotide core sequence.
LEU3 of Saccharomyces cerevisiae encodes an 886-amino-acid polypeptide that regulates transcription of a group of genes involved in leucine biosynthesis and has been shown to bind specifically to a 114-base-pair DNA fragment of the LEU2 upstream region (P. Friden and P. Schimmel, Mol. Cell. Biol. 7:2707-2717, 1987). We show here that, in addition to LEU2, LEU3 binds in vitro to sequences in the promoter regions of LEU1, LEU4, ILV2, and, by inference, ILV5. The largely conserved decanucleotide core sequence shared by the binding sites in these genes is CCGGNNCCGG. Methylation interference footprinting experiments show that LEU3 makes symmetrical contacts with the conserved bases that lie in the major groove. Synthetic oligonucleotides (19 to 29 base pairs) which contain the core decanucleotide and flanking sequences of LEU1, LEU2, LEU4, and ILV2 have individually been placed upstream of a LEU3-insensitive test promoter. The expression of each construction is activated by LEU3, although the degree of activation varies considerably according to the specific oligonucleotide which is introduced. A promoter construction with substitutions in the core sequence remains LEU3 insensitive, however. One of the oligonucleotides (based on a LEU2 sequence) was also tested and shown to confer leucine-sensitive expression on the test promoter. The results demonstrate that only a short sequence element is necessary for LEU3-dependent promoter binding and activation and provide direct evidence for an expanded repertoire of genes that are activated by LEU3.


Cardiovascular responses to pulmonary artery catheterization.
This study compares prospectively the cardiovascular and catecholamine responses to central venous and pulmonary artery catheterization before and after induction of general anaesthesia. Twenty patients for elective coronary artery surgery were randomized into two groups. One group had central venous and pulmonary artery catheterization performed awake using local anaesthesia. The other group had these catheters inserted following induction of general anesthesia. In all patients heart rate, arterial blood pressure, ST segment analysis and epinephrine and norepinephrine levels were measured prior to central venous cannulation and at 2-min intervals until placement of the lines was achieved. There were no statistically significant changes in any cardiovascular or catecholamine variable with time compared with the base-line measurements. There were no statistically significant differences in plasma catecholamine levels between the awake and the anaesthetized groups.


Molecular cloning and characterization of endoplasmic reticulum stress related genes grp78 and atf6α from black seabream (Acanthopagrus schlegelii) and their expressions in response to nutritional regulation.
Glucose-regulated protein 78 (grp78) and activating transcription factor 6α (atf6α) are considered vital endoplasmic reticulum (ER) molecular chaperones and ER stress (ERS) sensors, respectively. In the present study, the full cDNA sequences of these two ERS-related genes were first cloned and characterized from black seabream (Acanthopagrus schlegelii). The grp78 cDNA sequence is 2606 base pair (bp) encoding a protein of 654 amino acids (aa). The atf6α cDNA sequence is 2168 base pair (bp) encoding a protein of 645 aa. The predicted aa sequences of A. schlegelii grp78 and atf6α indicated that the proteins contain all the structural features, which were characteristic of the two genes in other species. Tissues transcript abundance analysis revealed that the mRNAs of grp78 and atf6α were expressed in all measured tissues, but the highest expression of these two genes was all recorded in the gill followed by liver/ brain. Moreover, in vivo experiment found that fish intake of a high lipid diet (HLD) can trigger ERS by activating grp78/Grp78 and atf6α/Atf6α. However, it can be alleviated by dietary betaine supplementation, similar results were also obtained by in vitro experiment using primary hepatocytes of A. schlegelii. These findings will be beneficial for us to evaluate the regulator effects of HLD supplemented with betaine on ERS at the molecular level, and thus provide some novel insights into the functions of betaine in marine fish fed with an HLD.


Cathepsin B Is Inhibited in Mutant Cells Selected during Persistent Reovirus Infection.
Persistent reovirus infections of murine L929 (L) fibroblast cells select mutant (LX) cells that do not support proteolytic disassembly of reovirus virions within the endocytic pathway. To better understand the function and regulation of endocytic proteases, we conducted experiments to define the block to reovirus disassembly displayed by LX cells. In contrast to parental L cells, mutant LX cells harbor defects that interfere with the maturation and activity of cathepsin B and cathepsin L but not cathepsin H. The cDNAs encoding cathepsin B and cathepsin L in L cells are identical to those in LX cells, indicating that LX cells manifest an extrinsic block to the function of these enzymes. Mixed lysates of L cells and LX cells lack activity of both cathepsin B and cathepsin L, suggesting the presence of an inhibitor of cathepsin function in LX cells. A cathepsin B-green fluorescent protein (GFP) fusion protein expressed in L cells and purified by immunoprecipitation retains cathepsin B activity, whereas cathepsin B-GFP expressed in LX cells does not. However, activity of cathepsin B-GFP expressed in LX cells can be recovered by incubating the immunoprecipitate with L cell lysate followed by immunoprecipitation, providing further evidence that LX cells express a cathepsin inhibitor. Native-gel electrophoresis and gel filtration chromatography demonstrate that, in both cell lines, the double-chain form of cathepsin B is sequestered in a large molecular weight complex that renders this form of the enzyme inactive. Alteration of this sequestration complex appears to be responsible for inhibition of cathepsin B in LX cells. These findings suggest that cathepsins can be regulated within the endocytic pathway. Moreover, this regulation influences host cell susceptibility to intracellular pathogens.


Familial adenomatous polyposis: ileo-anal pouch versus ileo-rectal anastomosis.
In this study we describe the presentation, treatment, and complications of 27 FAP patients. Treatment of Familial adenomatous polyposis (FAP) is centered on early recognition and curative surgery with either restorative proctocolectomy with ileal-pouch-anal-anastomosis (IPAA) or colectomy with ileo-rectal anastomosis (IRA). All patients diagnosed with FAP at our center from 2008 to 2012 were included in this case series. Either IPAA or IRA was used for treatment. Complications were recorded for 12 months after the procedure. Overall 27 patients were included, 12 (44.44%) index patients, and 15 (55.55%) relatives diagnosed by screening. Eight Index patients presented with rectal bleeding, two with occult fecal blood and two with abdominal masses found to be desmoid tumors. Nineteen patients were treated by IPAA, 6 with IRA, and 2 were inoperable due to diffuse desmoid tumors. Daytime stool frequency was the most common side effect (70.37%), followed by bowel discomfort episodes (55.56%), requiring dietary restrictions (37.4%), passive incontinence (25.93%), soiling (22.22%), nighttime stool frequency (18.52%), flatus incontinence (16.0%), and anastomosis leakage (3.70%). On average patients treated by IPAA experienced less complication than those treated by IRA. compared with previous reports, this series had older age of diagnosis, higher rate of adenocarcinoma at diagnosis, and fewer side effects after IPAA than IRA. The latter may reflect technique improvement with experience, and if supported by future studies, will cement IPAA as the treatment of choice in FAP.


CRISPR-Cas9 and the Promise of a Better Future.
Due to its simplicity, low cost and accuracy, CRISPR-Cas9 has become a promising new technique in the field of gene editing. However, despite its virtues, it is not yet immune to scientific hazards and ethical legal concerns. These concerns have been used to justify opposition to genetic manipulation, and have led to some regulations to ban or impose a moratorium based on the precautionary principle. In Europe, regulation mostly comes from the European Union and the Council of Europe, both very cautious towards gene editing. In this article, two arguments on the future legal framework of CRISPR-Cas9 are made. The first is that continued research will contribute to more scientific accuracy; thus, the precautionary principle should promote regulated research to achieve this aim. The second is that most of the legal and ethical concerns surrounding CRISPR-Cas9 are based on unfounded prejudice emanating from a mystical understanding of the human genome.


Assessment of arterial stiffness for clinical and epidemiological studies: methodological considerations for validation and entry into the European Renal and Cardiovascular Medicine registry.
Epidemiological studies have highlighted the role of arterial stiffness as a risk factor for development of cardiovascular (CV) diseases. Moreover, aortic stiffness has been shown to be a significant predictive factor of all-cause and CV mortality in different populations including patients with end-stage renal disease. Pulse-wave velocity (PWV) is the most widely used technique to assess arterial stiffness. Although PWV can be measured on any artery or between any arterial sites, only carotid-to-femoral PWV, representing stiffness of the aorta and iliofemoral axes, has been shown to have predictive value for morbidity and mortality. The several available commercial devices differ according to the type of signal (pressure, distension, flow) or by recording both sites simultaneously or using ECG synchronization. It is also possible to directly measure arterial diameter changes during the cardiac cycle and link them to local pulse-pressure changes, which provides the pressure-diameter relationship and stress-strain relationship if arterial wall thickness is also measured. These techniques are based on high-precision vascular echo tracking or magnetic resonance imaging and applanation tonometry. This paper summarizes the basic principles of arterial haemodynamics and various methodologies to assess stiffness and the latest consensus recommendations for clinical applications.