Population Pharmacokinetic Modeling as a Tool To Characterize the Decrease in Ciprofloxacin Free Interstitial Levels Caused by Pseudomonas aeruginosa Biofilm Lung Infection in Wistar Rats.
Biofilm formation plays an important role in the persistence of pulmonary infections, for example, in cystic fibrosis patients. So far, little is known about the antimicrobial lung disposition in biofilm-associated pneumonia. This study aimed to evaluate, by microdialysis, ciprofloxacin (CIP) penetration into the lungs of healthy and Pseudomonas aeruginosa biofilm-infected rats and to develop a comprehensive model to describe the CIP disposition under both conditions. P. aeruginosa was immobilized into alginate beads and intratracheally inoculated 14 days before CIP administration (20 mg/kg of body weight). Plasma and microdialysate were sampled from different animal groups, and the observations were evaluated by noncompartmental analysis (NCA) and population pharmacokinetic (popPK) analysis. The final model that successfully described all data consisted of an arterial and a venous central compartment and two peripheral distribution compartments, and the disposition in the lung was modeled as a two-compartment model structure linked to the venous compartment. Plasma clearance was approximately 32% lower in infected animals, leading to a significantly higher level of plasma CIP exposure (area under the concentration-time curve from time zero to infinity, 27.3 ± 12.1 μg · h/ml and 13.3 ± 3.5 μg · h/ml in infected and healthy rats, respectively). Despite the plasma exposure, infected animals showed a four times lower tissue concentration/plasma concentration ratio (lung penetration factor = 0.44 and 1.69 in infected and healthy rats, respectively), and lung clearance (CLlung) was added to the model for these animals (CLlung = 0.643 liters/h/kg) to explain the lower tissue concentrations. Our results indicate that P. aeruginosa biofilm infection reduces the CIP free interstitial lung concentrations and increases plasma exposure, suggesting that plasma concentrations alone are not a good surrogate of lung concentrations.


Redox-dependent complex formation by an ATP-dependent activator of the corrinoid/iron-sulfur protein.
Movement, cell division, protein biosynthesis, electron transfer against an electrochemical gradient, and many more processes depend on energy conversions coupled to the hydrolysis of ATP. The reduction of metal sites with low reduction potentials (E(0') < -500 mV) is possible by connecting an energetical uphill electron transfer with the hydrolysis of ATP. The corrinoid-iron/sulfur protein (CoFeSP) operates within the reductive acetyl-CoA pathway by transferring a methyl group from methyltetrahydrofolate bound to a methyltransferase to the [Ni-Ni-Fe(4)S(4)] cluster of acetyl-CoA synthase. Methylation of CoFeSP only occurs in the low-potential Co(I) state, which can be sporadically oxidized to the inactive Co(II) state, making its reductive reactivation necessary. Here we show that an open-reading frame proximal to the structural genes of CoFeSP encodes an ATP-dependent reductive activator of CoFeSP. Our biochemical and structural analysis uncovers a unique type of reductive activator distinct from the electron-transferring ATPases found to reduce the MoFe-nitrogenase and 2-hydroxyacyl-CoA dehydratases. The CoFeSP activator contains an ASKHA domain (acetate and sugar kinases, Hsp70, and actin) harboring the ATP-binding site, which is also present in the activator of 2-hydroxyacyl-CoA dehydratases and a ferredoxin-like [2Fe-2S] cluster domain acting as electron donor. Complex formation between CoFeSP and its activator depends on the oxidation state of CoFeSP, which provides evidence for a unique strategy to achieve unidirectional electron transfer between two redox proteins.


Development of a New International Antiphospholipid Syndrome Classification Criteria Phase I/II Report: Generation and Reduction of Candidate Criteria.
An international multidisciplinary initiative, jointly supported by the American College of Rheumatology and European Alliance of Associations for Rheumatology, is underway to develop new rigorous classification criteria to identify patients with high likelihood of antiphospholipid syndrome (APS) for research purposes. The present study was undertaken to apply an evidence- and consensus-based approach to identify candidate criteria and develop a hierarchical organization of criteria within domains. During phase I, the APS classification criteria steering committee used systematic literature reviews and surveys of international APS physician scientists to generate a comprehensive list of items related to APS. In phase II, we reviewed the literature, administered surveys, formed domain subcommittees, and used Delphi exercises and nominal group technique to reduce potential APS candidate criteria. Candidate criteria were hierarchically organized into clinical and laboratory domains. Phase I generated 152 candidate criteria, expanded to 261 items with the addition of subgroups and candidate criteria with potential negative weights. Using iterative item reduction techniques in phase II, we initially reduced these items to 64 potential candidate criteria organized into 10 clinical and laboratory domains. Subsequent item reduction methods resulted in 27 candidate criteria, hierarchically organized into 6 additive domains (laboratory, macrovascular, microvascular, obstetric, cardiac, and hematologic) for APS classification. Using data- and consensus-driven methodology, we identified 27 APS candidate criteria in 6 clinical or laboratory domains. In the next phase, the proposed candidate criteria will be used for real-world case collection and further refined, organized, and weighted to determine an aggregate score and threshold for APS classification.


The impact of de-epithelialization of the amniotic membrane matrix on morphology of cultured human limbal epithelial cells subject to eye bank storage.
To investigate whether human limbal epithelial cells (HLEC) that have been cultured on intact or de-epithelialized amniotic membranes (AMs) demonstrate differences in morphology after 1 week of eye bank storage. HLEC were cultured from limbal explants for 3 weeks on intact AM and AM deprived of the amniotic epithelial cells by incubation with 0.02% ethylene diamine tetra acetic acid followed by mechanical scraping. The HLEC cultures were stored for 1 week in a closed container in a serum-based medium at 23°C. The surface morphology was assessed using scanning electron microscopy, and a quantitative comparison of desmosome and hemidesmosome numbers was performed using transmission electron microscopy. Although most superficial epithelial cells were closely attached to each other, with tightly opposed cell junctions and distinct cell borders, there was evidence of some cell separation in HLEC that had been cultured on intact and denuded AM after 1 week of storage. In both experimental groups, the epithelia were well stratified, consisting of basal column-shaped cells, suprabasal cuboid wing cells, and flat squamous superficial cells, but dilated intercellular spaces were observed. The total number of desmosomes per micron was 1.39 ± 0.77 in HLEC cultured on intact AM versus 0.98 ± 0.45 in HLEC expanded on denuded AM (P = 0.76). The total number of hemidesmosomes per micron in HLEC cultured on intact AM and denuded AM was 0.87 ± 0.34 and 0.78 ± 0.31, respectively (P = 0.70). Denuding of AM does not improve the structural integrity of cultured HLEC after eye bank storage.


Statistical power in stuttering research: a tutorial.
The capacity to make reliable inductive statements about populations is critical for the advancement of scientific knowledge. An important contribution to that advancement of knowledge is determining that effects are either present or not present in populations. Statistical power is an important methodological qualification for any research that presents statistical results, and particularly so for research that presents null results. In this paper we describe the statistical concept of power, outline parameters of research that influence it, and demonstrate how it is calculated. With reference to selected published research, attention is drawn to the problems associated with a body of underpowered research, one being that population effects may go undetected. One way to prevent this problem is to calculate power a priori in planning research and include confidence intervals when presenting the results of research. However, it is difficult, if not impossible in many cases, to obtain high participant numbers for communication disorders of low prevalence such as stuttering. With this in mind, the paper concludes with an attempt to open discussion about ways to redress the problems associated with statistical power.


Cost-utility and budget impact analysis of adding-on apremilast to biologic therapy in the treatment of moderate to severe plaque psoriasis, an Iranian payer perspective.
Plaque psoriasis is a chronic inflammatory disease with skin manifestations that affect the patients' quality of life negatively. The prevalence of psoriasis is approximately 2-3% worldwide and appears to be still on the increase. Due to the stigma problems, psoriasis has a significant effect on one's life that is often overlooked. The current study aimed to conduct the cost-utility evaluation and budget impact analysis of adding-on apremilast ahead of biologic therapy in the treatment of moderate to severe plaque psoriasis. The psoriatic patients who did not undergo the conventional systemic therapy were eligible to enter the defined sequences. An excel-based Markov model with 40 cycles of 3 months, each of which was adopted to compare the outcomes of each exclusively administered sequence in the treatment of moderate to severe plaque psoriasis. Two exclusive therapeutic sequences were considered. In the first sequence, apremilast was followed by biologics and in the second one, biologics were administered initially without apremilast. The results were extrapolated up to 10 years. The designed Markov model was also used in budget impact analysis. The cost-saving potential of the new treatment was accounted for the next 5 years. Incremental cost and incremental effect were reported in the base case scenario. Using the sequence consisting apremilast provided an additional 0.10 quality-adjusted life years and decreased total costs by about 11,100 USD per patient. These results were in line with the findings from sensitivity analysis. The cost-saving over 5 years is estimated to be around 30 million dollars for the Iran market following the use of the new treatment. In the treatment of moderate to severe plaque psoriasis, apremilast supplementation prior to biological treatments is more cost-effective than biological treatment alone.


In hot and cold water: differential life-history traits are key to success in contrasting thermal deep-sea environments.
Few species of reptant decapod crustaceans thrive in the cold-stenothermal waters of the Southern Ocean. However, abundant populations of a new species of anomuran crab, Kiwa tyleri, occur at hydrothermal vent fields on the East Scotia Ridge. As a result of local thermal conditions at the vents, these crabs are not restricted by the physiological limits that otherwise exclude reptant decapods south of the polar front. We reveal the adult life history of this species by piecing together variation in microdistribution, body size frequency, sex ratio, and ovarian and embryonic development, which indicates a pattern in the distribution of female Kiwaidae in relation to their reproductive development. High-density 'Kiwa' assemblages observed in close proximity to sources of vent fluids are constrained by the thermal limit of elevated temperatures and the availability of resources for chemosynthetic nutrition. Although adult Kiwaidae depend on epibiotic chemosynthetic bacteria for nutrition, females move offsite after extrusion of their eggs to protect brooding embryos from the chemically harsh, thermally fluctuating vent environment. Consequently, brooding females in the periphery of the vent field are in turn restricted by low-temperature physiological boundaries of the deep-water Southern Ocean environment. Females have a high reproductive investment in few, large, yolky eggs, facilitating full lecithotrophy, with the release of larvae prolonged, and asynchronous. After embryos are released, larvae are reliant on locating isolated active areas of hydrothermal flow in order to settle and survive as chemosynthetic adults. Where the cold water restricts the ability of all adult stages to migrate over long distances, these low temperatures may facilitate the larvae in the location of vent sites by extending the larval development period through hypometabolism. These differential life-history adaptations to contrasting thermal environments lead to a disjunct life history among males and females of K. tyleri, which is key to their success in the Southern Ocean vent environment. We highlight the complexity in understanding the importance of life-history biology, in combination with environmental, ecological and physiological factors contributing to the overall global distribution of vent-endemic species.


Genetic analysis of the blood transcriptome of young healthy pigs to improve disease resilience.
Disease resilience is the ability of an animal to maintain productive performance under disease conditions and is an important selection target. In pig breeding programs, disease resilience must be evaluated on selection candidates without exposing them to disease. To identify potential genetic indicators for disease resilience that can be measured on selection candidates, we focused on the blood transcriptome of 1594 young healthy pigs with subsequent records on disease resilience. Transcriptome data were obtained by 3'mRNA sequencing and genotype data were from a 650 K genotyping array. Heritabilities of the expression of 16,545 genes were estimated, of which 5665 genes showed significant estimates of heritability (p < 0.05), ranging from 0.05 to 0.90, with or without accounting for white blood cell composition. Genes with heritable expression levels were spread across chromosomes, but were enriched in the swine leukocyte antigen region (average estimate > 0.2). The correlation of heritability estimates with the corresponding estimates obtained for genes expressed in human blood was weak but a sizable number of genes with heritable expression levels overlapped. Genes with heritable expression levels were significantly enriched for biological processes such as cell activation, immune system process, stress response, and leukocyte activation, and were involved in various disease annotations such as RNA virus infection, including SARS-Cov2, as well as liver disease, and inflammation. To estimate genetic correlations with disease resilience, 3205 genotyped pigs, including the 1594 pigs with transcriptome data, were evaluated for disease resilience following their exposure to a natural polymicrobial disease challenge. Significant genetic correlations (p < 0.05) were observed with all resilience phenotypes, although few exceeded expected false discovery rates. Enrichment analysis of genes ranked by estimates of genetic correlations with resilience phenotypes revealed significance for biological processes such as regulation of cytokines, including interleukins and interferons, and chaperone mediated protein folding. These results suggest that expression levels in the blood of young healthy pigs for genes in biological pathways related to immunity and endoplasmic reticulum stress have potential to be used as genetic indicator traits to select for disease resilience.


Vagus nerve stimulation for children with treatment-resistant epilepsy: a consecutive series of 141 cases.
The authors undertook this study to analyze the efficacy of vagus nerve stimulation (VNS) in a large consecutive series of children 18 years of age and younger with treatment-resistant epilepsy and compare the safety and efficacy in children under 12 years of age with the outcomes in older children. The authors retrospectively reviewed 141 consecutive cases involving children (75 girls and 66 boys) with treatment-resistant epilepsy in whom primary VNS implantation was performed by the senior author between November 1997 and April 2008 and who had at least 1 year of follow-up since implantation. The patients' mean age at vagus nerve stimulator insertion was 11.1 years (range 1-18 years). Eighty-six children (61.0%) were younger than 12 years at time of VNS insertion (which constitutes off-label usage of this device). Follow-up was complete for 91.8% of patients and the mean duration of VNS therapy in these patients was 5.2 years (range 25 days-11.4 years). Seizure frequency significantly improved with VNS therapy (mean reduction 58.9%, p < 0.0001) without a significant reduction in antiepileptic medication burden (median number of antiepileptic drugs taken 3, unchanged). Reduction in seizure frequency of at least 50% occurred in 64.8% of patients and 41.4% of patients experienced at least a 75% reduction. Major (3) and minor (6) complications occurred in 9 patients (6.4%) and included 1 deep infection requiring device removal, 1 pneumothorax, 2 superficial infections treated with antibiotics, 1 seroma/hematoma treated with aspiration, persistent cough in 1 patient, severe but transient neck pain in 1 patient, and hoarseness in 2 patients. There was no difference in efficacy or complications between children 12 years of age and older (FDA-approved indication) and those younger than 12 years of age (off-label usage). Linear regression analyses did not identify any demographic and clinical variables that predicted response to VNS. Vagus nerve stimulation is a safe and effective treatment for treatment-resistant epilepsy in young adults and children. Over 50% of patients experienced at least 50% reduction in seizure burden. Children younger than 12 years had a response similar to that of older children with no increase in complications. Given the efficacy of this device and the devastating effects of persistent epilepsy during critical developmental epochs, randomized trials are needed to potentially expand the indications for VNS to include younger children.


Intermediate class of mRNAs in African swine fever virus.
A transcriptional analysis of the African swine fever virus (ASFV) I226R and I243L genes is presented. Steady-state kinetics and transfection experiments showed the existence of a new temporal class of ASFV mRNAs transcribed from these genes, with the characteristics of the poxvirus intermediate transcripts. Transcription of the I226R gene gave rise to intermediate and late mRNAs that started from different sites, while the I243L gene produced early, intermediate, and late mRNAs, also starting from different sites. The presence of intermediate genes suggests a cascade model for the regulation of ASFV gene expression.


Proteinuria selectivity index in renal disease.
One of the main barriers to early detection and subsequent prevention of kidney diseases is the accessibility and feasibility of testing, especially in urine research. The proteinuria selectivity index (PSI or SI) is a method used to assess changes in glomerular permeability in glomerular diseases. It describes the pattern of proteinuria by comparing the clearance rates of large molecular proteins and transferrin, categorizing it as selective or non-selective. PSI is widely applied for kidney disease classification, prediction of corticosteroid efficacy, and prognosis. Herein, we reviewed the clinical applications and recent advancements of PSI in glomerular diseases, compared it with commonly used renal function biomarkers, and discussed the future research directions for PSI as a potential predictive marker for response to specific biologics.


TERT Promoter Mutations Increase Sense and Antisense Transcription from the TERT Promoter.
Background: Chief among mechanisms of telomerase reverse transcriptase (TERT) reactivation is the appearance of mutations in the TERT promoter. The two main TERT promoter mutations are C>T transitions located -146C>T and -124C>T upstream from the translational start site. They generate a novel Ets/TCF binding site. Both mutations are mutually exclusive and -124C>T is strikingly overrepresented in most cancers. We investigated whether this mutational bias and mutual exclusion could be due to transcriptional constraints. Methods: We compared sense and antisense transcription of a panel of TERT promoter-luciferase vectors harboring the -124C>T and -146C>T mutations alone or together. lncRNA TAPAS levels were measured by RT-PCR. Results: Both mutations generally increased TERT transcription by 2-4-fold regardless of upstream and downstream regulatory elements. The double mutant increased transcription in an additive fashion, arguing against a direct transcriptional constraint. The -146C>T mutation, alone or in combination with -124C>T, also unleashed antisense transcription. In line with this finding, lncRNA TAPAS was higher in cells with mutated TERT promoter (T98G and U87) than in cells with wild-type promoter, suggesting that lncRNA TAPAS may balance the effect of TERT promoter mutations. Conclusions: -146C>T and -124C>T TERT promoter mutations increase TERT sense and antisense transcription, and the double mutant features higher transcription levels. Increased antisense transcription may contain TERT expression within sustainable levels.


Structural mechanism for ubiquitinated-cargo recognition by the Golgi-localized, gamma-ear-containing, ADP-ribosylation-factor-binding proteins.
The Golgi-localized, gamma-ear-containing, Arf (ADP-ribosylation factor)-binding (GGA) proteins are clathrin adaptors that mediate the sorting of transmembrane-cargo molecules at the trans-Golgi network and endosomes. Cargo proteins can be directed into the GGA pathway by at least two different types of sorting signals: acidic cluster-dileucine motifs and covalent modification by ubiquitin. The latter modification is recognized by the GGAs through binding to their GAT [GGA and TOM (target of Myb)] domain. Here we report the crystal structure of the GAT domain of human GGA3 in a 1:1 complex with ubiquitin at 2.8-A resolution. Ubiquitin binds to a hydrophobic and acidic patch on helices alpha1 and alpha2 of the GAT three-helix bundle that includes Asn-223, Leu-227, Glu-230, Met-231, Asp-244, Glu-246, Leu-247, Glu-250, and Leu-251. The GAT-binding surface on ubiquitin is a hydrophobic patch centered on Ile-44 that is also responsible for binding most other ubiquitin effectors. The ubiquitin-binding site observed in the crystal is distinct from the Rabaptin-5-binding site on helices alpha2 and alpha3 of the GAT domain. Mutational analysis and modeling of the ubiquitin-Rabaptin-5-GAT ternary complex indicates that ubiquitin and Rabaptin-5 can bind to the GAT domain at two different sites without any steric conflict. This ability highlights the GAT domain as a hub for interactions with multiple partners in trafficking.


Related Factors of Cerebral Hemorrhage after Cerebral Infarction and the Effect of Atorvastatin Combined with Intensive Nursing Care.
Cerebral infarction is a common neurological disease with high incidence, which is the main factor causing death and disability in adults in China. Cerebral hemorrhage transformation is a common clinical complication. High NIHSS score at admission, atrial fibrillation, and small artery occlusion cerebral infarction can increase the risk of cerebral infarction complicated with hemorrhage transformation. To explore the related factors of cerebral hemorrhage transformation after cerebral infarction and the value of atorvastatin calcium tablets combined with early intensive care measures. In this study, a case-control study was conducted. Sixty patients with hemorrhagic transformation after cerebral infarction admitted to the Department of Neurology of our hospital from January 2017 to June 2021 were selected as the observation group, and 90 patients with cerebral infarction without hemorrhagic transformation during the same period were selected as the control group. The risk factors of hemorrhagic transformation after cerebral infarction were analyzed. The results of logistic regression model showed that the increased National Institutes of Health Stroke Scale (NIHSS) score at admission, hypertension, atrial fibrillation, TOAST classification of small artery occlusion, and large infarction lesions were the risk factors for hemorrhagic transformation in patients with cerebral infarction (P < 0.05). After 2 weeks and 4 weeks of treatment, the NIHSS scores of the intervention group were lower than those of the conventional group (P < 0.05). NIHSS scores of the two groups after treatment were significantly lower than those before treatment (P < 0.05). After three months of treatment, the patients in the intervention group with GOS score of 5 points accounted for 16.67%, and the patients with GOS score of 4 points accounted for 56.67%. The patients in the conventional group with GOS score of 5 points accounted for 6.67%, and the patients with GOS score of 4 points accounted for 33.33%. The prognosis of the intervention group was better than that of the conventional group on the whole (P < 0.05). Patients with hypertension, large infarction lesions, high NIHSS score at admission, atrial fibrillation, and small artery occlusion cerebral infarction can increase the risk of bleeding transformation in patients with cerebral infarction. For patients with bleeding transformation, atorvastatin calcium tablets combined with early intensive nursing intervention has a certain value for improving the prognosis of patients.


[Percentage of epithelial cells, fibroblasts and endothelial cells in the dog thyroid].
The proportions of various cells types were studied in the thyroids of a variety of dogs used for laboratory experiments. A remarkable constancy was found with epithelial cells accounting for 70% of the total number of cells. The relative proportions of luminal and epithelial cells spaces were also very similar in different animals (47% and 26% respectively).


Photoluminescence and Nonlinear Optical Properties of Two Terpyridine-Based Hybrid Zn/Cd Halides.
Hybrid metal halides (HMHs) with low-dimensional structures have attracted increasing attention due to their striking optical properties. Herein, two new zero-dimensional HMHs have been fabricated by CdCl2/ZnCl2 and 4'-(4-pyridyl-phenyl)-2,2':6',2″-terpyridine (Tpy), including (TpyH3)[CdCl4][Cl] (Tpy-Cd) and (TpyH3)[ZnCl4][Cl] (Tpy-Zn). Their structures are consisted of a [TpyH3]3+ organic cation, an inorganic [ZnCl4] or [CdCl4] tetrahedron, and one isolated Cl- anion. Tpy-Cd crystallizes to a noncentrosymmetric structure and possesses a moderate second harmonic response of 0.72 × KH2PO4, while Tpy-Zn features a centrosymmetric space group. Though Tpy-Cd and Tpy-Zn crystallize into space groups of completely different symmetry due to distinct connection mode and molecular distortion, they display quite similar photoluminescence of bright green light emission under ultraviolet excitation, nearly identical in Stokes shift, photoluminescence quantum yield, decay lifetime, and energy. The photoluminescence quantum yields of green light emission were measured to be nearly 25%, outperforming most of the Cd/Zn low-dimensional HMHs.


Histologic grade is correlated with outcome after resection of hepatic neuroendocrine neoplasms.
The behavior of neuroendocrine neoplasms is poorly defined, and predictors of outcome after surgical resection have yet to be identified. Consequently, guidelines for treatment remain unclear. Current pathologic classification systems do not permit meaningful discrimination of hepatic neuroendocrine neoplasms. The authors reviewed prospectively maintained databases from 2 institutions of patients who underwent hepatic resection for neuroendocrine neoplasms between 1990 and 2006. Patient, tumor, and operative characteristics were analyzed to identify factors associated with overall survival, progression-free survival, and symptom control. Hepatic neoplasms were stratified by using a 3-tier pathologic classification system based on the number of mitotic figures and the presence of tumor necrosis that was recently validated for pancreatic neuroendocrine neoplasms. Seventy patients were identified from the databases. Low-grade, intermediate-grade, and high-grade neoplasms were identified in 53%, 37%, and 10% of patients, respectively. After a median follow-up of 51 months, the median overall survival for all patients was 91 months, and it was 108 months when 7 patients with high-grade neuroendocrine carcinomas were excluded. Progressive disease was eventually observed in 81% of patients, and the median progression-free survival was 17 months. The median time to the onset of symptoms was 39 months for patients who presented with hormonal symptoms and 80 months for all patients. Histologic grade was associated with poorer overall and progression-free survival. When performed in a context of aggressive multimodality therapy, long-term outcomes after partial hepatectomy for hepatic neuroendocrine neoplasms were favorable; however, disease progression was eventually observed in the majority of patients. Several oncologic variables were associated with significant differences in survival after resection. A novel pathologic classification system appears to enhance prognostic stratification of patients with hepatic neuroendocrine neoplasms.


Classification of complications of epilepsy surgery and invasive diagnostic procedures: A proposed protocol and feasibility study.
In epilepsy surgery, which aims to treat seizures and thereby to improve the lives of persons with drug-resistant epilepsy, the chances of attaining seizure relief must be carefully weighed against the risks of complications and expected adverse events. The interpretation of data regarding complications of epilepsy surgery and invasive diagnostic procedures is hampered by a lack of uniform definitions and method of data collection. Based on a review of previous definitions and classifications of complications, we developed a proposal for a new classification. This proposal was then subject to revisions after expert opinion within E-pilepsy, an EU-funded European pilot network of reference centers in refractory epilepsy and epilepsy surgery, later incorporated into the ERN (European Reference Network) EpiCARE. This version was discussed with recognized experts, and a final protocol was agreed to after further revision. The final protocol was evaluated in practical use over 1 year in three of the participating centers. One hundred seventy-four consecutive procedures were included with 35 reported complications. This report presents a multidimensional classification of complications in epilepsy surgery and invasive diagnostic procedures, where complications are characterized in terms of their immediate effects, resulting permanent symptoms, and consequences on activities of daily living. We propose that the protocol will be helpful in the work to promote safety in epilepsy surgery and for future studies designed to identify risk factors for complications. Further work is needed to address the reporting of outcomes as regards neuropsychological function, activities of daily living, and quality of life.


Local flap reconstruction of large scalp defects.
Scalp defects can have a number of origins, and their repair is dependent upon their location, size and depth. In the case of the scalp, the repair of even small defects is complicated. Local flaps are the reference for the reconstruction of such defects. Knowledge of scalp anatomy is essential for preparing these flaps, which must be based on one or two vascular pedicles to afford a large rotation angle--thereby facilitating closure of the defect. The parietal zone is the location offering the greatest flap mobilization possibilities. We present a case involving the repair of a major pericranial frontoparietal scalp defect. A local transverse posterior transpositioning scalp flap was raised with the posterior auricular and occipital arteries as vascular pedicle. Following repositioning of the flap, a free partial-thickness skin graft from the thigh was used to cover the donor zone. A review is provided of the different techniques for the reconstruction of large scalp defects.


Engineering a change in metal-ion specificity of the iron-dependent superoxide dismutase from Mycobacterium tuberculosis-- X-ray structure analysis of site-directed mutants.
We have refined the X-ray structures of two site-directed mutants of the iron-dependent superoxide dismutase (SOD) from Mycobacterium tuberculosis. These mutations which affect residue 145 in the enzyme (H145Q and H145E) were designed to alter its metal-ion specificity. This residue is either Gln or His in homologous SOD enzymes and has previously been shown to play a role in active-site interactions since its side-chain helps to coordinate the metal ion via a solvent molecule which is thought to be a hydroxide ion. The mutations were based on the observation that in the closely homologous manganese dependent SOD from Mycobacterium leprae, the only significant difference from the M. tuberculosis SOD within 10 A of the metal-binding site is the substitution of Gln for His at position 145. Hence an H145Q mutant of the M. tuberculosis (TB) SOD was engineered to investigate this residue's role in metal ion dependence and an isosteric H145E mutant was also expressed. The X-ray structures of the H145Q and H145E mutants have been solved at resolutions of 4.0 A and 2.5 A, respectively, confirming that neither mutation has any gross effects on the conformation of the enzyme or the structure of the active site. The residue substitutions are accommodated in the enzyme's three-dimensional structure by small local conformational changes. Peroxide inhibition experiments and atomic absorption spectroscopy establish surprisingly the H145E mutant SOD has manganese bound to it whereas the H145Q mutant SOD retains iron as the active-site metal. This alteration in metal specificity may reflect on the preference of manganese ions for anionic ligands.


Porcine Ischemic Wound-Healing Model for Preclinical Testing of Degradable Biomaterials.
Impaired wound healing that mimics chronic human skin pathologies is difficult to achieve in current animal models, hindering testing and development of new therapeutic biomaterials that promote wound healing. In this article, we describe a refinement and simplification of the porcine ischemic wound model that increases the size and number of experimental sites per animal. By comparing three flap geometries, we adopted a superior configuration (15 × 10 cm) that enabled testing of twenty 1 cm2 wounds in each animal: 8 total ischemic wounds within 4 bipedicle flaps and 12 nonischemic wounds. The ischemic wounds exhibited impaired skin perfusion for ∼1 week. To demonstrate the utility of the model for comparative testing of tissue regenerative biomaterials, we evaluated the healing process in wounds implanted with highly porous poly (thioketal) urethane (PTK-UR) scaffolds that were fabricated through reaction of reactive oxygen species (ROS)-cleavable PTK macrodiols with isocyanates. PTK-lysine triisocyanate (LTI) scaffolds degraded significantly in vitro under both oxidative and hydrolytic conditions whereas PTK-hexamethylene diisocyanate trimer (HDIt) scaffolds were resistant to hydrolytic breakdown and degraded exclusively through an ROS-dependent mechanism. Upon placement into porcine wounds, both types of PTK-UR materials fostered new tissue ingrowth over 10 days in both ischemic and nonischemic tissue. However, wound perfusion, tissue infiltration and the abundance of pro-regenerative, M2-polarized macrophages were markedly lower in ischemic wounds independent of scaffold type. The PTK-LTI implants significantly improved tissue infiltration and perfusion compared with analogous PTK-HDIt scaffolds in ischemic wounds. Both LTI and HDIt-based PTK-UR implants enhanced M2 macrophage activity, and these cells were selectively localized at the scaffold/tissue interface. In sum, this modified porcine wound-healing model decreased animal usage, simplified procedures, and permitted a more robust evaluation of tissue engineering materials in preclinical wound healing research. Deployment of the model for a relevant biomaterial comparison yielded results that support the use of the PTK-LTI over the PTK-HDIt scaffold formulation for future advanced therapeutic studies.


Gamma Knife surgery for pituitary adenomas: factors related to radiological and endocrine outcomes.
Gamma Knife surgery (GKS) is a common treatment for recurrent or residual pituitary adenomas. This study evaluates a large cohort of patients with a pituitary adenoma to characterize factors related to endocrine remission, control of tumor growth, and development of pituitary deficiency. A total of 418 patients who underwent GKS with a minimum follow-up of 6 months (median 31 months) and for whom there was complete follow-up were evaluated. Statistical analysis was performed to evaluate for significant factors (p < 0.05) related to treatment outcomes. In patients with a secretory pituitary adenoma, the median time to endocrine remission was 48.9 months. The tumor margin radiation dose was inversely correlated with time to endocrine remission. Smaller adenoma volume correlated with improved endocrine remission in those with secretory adenomas. Cessation of pituitary suppressive medications at the time of GKS had a trend toward statistical significance in regard to influencing endocrine remission. In 90.3% of patients there was tumor control. A higher margin radiation dose significantly affected control of adenoma growth. New onset of a pituitary hormone deficiency following GKS was seen in 24.4% of patients. Treatment with pituitary hormone suppressive medication at the time of GKS, a prior craniotomy, and larger adenoma volume at the time of radiosurgery were significantly related to loss of pituitary function. Smaller adenoma volume improves the probability of endocrine remission and lowers the risk of new pituitary hormone deficiency with GKS. A higher margin dose offers a greater chance of endocrine remission and control of tumor growth.


[Application of the color Doppler scanning in dynamic follow up after the operation on thyroid gland].
After conduction of operation on thyroid gland (TG) 35 patients were examined using color Doppler mapping. The TG stump blood supply was determined mainly along periphery in sites of newly created membranes. Maximal rate of the TG tissue growth was observed in 1 year. The changes of the structure type can not be used as the test for establishing of the "goiter recurrency" diagnosis.


Broadband ultraviolet-visible transient absorption spectroscopy in the nanosecond to microsecond time domain with sub-nanosecond time resolution.
A combination of sub-nanosecond photoexcitation and femtosecond supercontinuum probing is used to extend femtosecond transient absorption spectroscopy into the nanosecond to microsecond time domain. Employing a passively Q-switched frequency tripled Nd:YAG laser and determining the jitter of the time delay between excitation and probe pulses with a high resolution time delay counter on a single-shot basis leads to a time resolution of 350 ps in picosecond excitation mode. The time overlap of almost an order of magnitude between fs and sub-ns excitation mode permits to extend ultrafast transient absorption (TA) experiments seamlessly into time ranges traditionally covered by laser flash photolysis. The broadband detection scheme eases the identification of intermediate reaction products which may remain undetected in single-wavelength detection flash photolysis arrangements. Single-shot referencing of the supercontinuum probe with two identical spectrometer/CCD arrangements yields an excellent signal-to-noise ratio for the so far investigated chromophores in short to moderate accumulation times.


Auxin transport through PIN-FORMED 3 (PIN3) controls shade avoidance and fitness during competition.
Plants grow in dense vegetations at the risk of being out-competed by neighbors. To increase their competitive power, plants display adaptive responses, such as rapid shoot elongation (shade avoidance) to consolidate light capture. These responses are induced upon detection of proximate neighbors through perception of the reduced ratio between red (R) and far-red (FR) light that is typical for dense vegetations. The plant hormone auxin is a central regulator of plant development and plasticity, but until now it has been unknown how auxin transport is controlled to regulate shade-avoidance responses. Here, we show that low R:FR detection changes the cellular location of the PIN-FORMED 3 (PIN3) protein, a regulator of auxin efflux, in Arabidopsis seedlings. As a result, auxin levels in the elongating hypocotyls are increased under low R:FR. Seedlings of the pin3-3 mutant lack this low R:FR-induced increase of endogenous auxin in the hypocotyl and, accordingly, have no elongation response to low R:FR. We hypothesize that low R:FR-induced stimulation of auxin biosynthesis drives the regulation of PIN3, thus allowing shade avoidance to occur. The adaptive significance of PIN3-mediated control of shade-avoidance is shown in plant competition studies. It was found that pin3 mutants are outcompeted by wild-type neighbors who suppress fitness of pin3-3 by 40%. We conclude that low R:FR modulates the auxin distribution by a change in the cellular location of PIN3, and that this control can be of great importance for plants growing in dense vegetations.


A possible murine model for investigation of pathogenesis of sudden infant death syndrome.
Several studies have indicated a possible causative role of toxigenic bacteria in sudden infant death syndrome (SIDS). This study examined the effect of toxigenic E. coli on pregnant and infant mice to determine if these animals could be used as a model for SIDS pathogenesis. Strains of E. coli from the intestinal contents of infants who have died of SIDS or other causes and from the faeces of healthy infants were collected over a broad time scale. The isolates were tested for their ability to produce then known toxins of E. coli and were serotyped (O and H antigens). Certain serotypes (e.g. O1:H- and O25:H1) emerged significantly more frequently from cases of SIDS than from healthy infants and isolates of these types were generally toxigenic in Vero-cell cultures but whose verotoxicity was not related to classical Shiga or other known toxins. This mouse model was developed to test the effects of these toxigenic and also non-toxigenic strains. Four apparently healthy pups aged between 17 and 21 days died unobserved overnight but no pups of the 54 control mice died suddenly (P = 0.0247, Fisher's exact test). These were considered to represent sudden unexpected deaths. Pathological effects compatible with those in SIDS were observed in mouse pups exposed to toxigenic strains indicating this model may be suitable for further study into the pathogenesis of unexpected deaths in infancy. Providing an animal model of SIDS would promote a much better avenue for studying the pathogenesis of this enigmatic condition.


"I thought that I had to be alive to repay my parents": Filial piety as a risk and protective factor for suicidal behavior in a qualitative study of Chinese women.
Filial piety involves the Confucian view that children always have a duty to be obedient and to provide care for their parents. Filial piety has been described as both a risk and a protective factor in depression and suicide. This qualitative study aimed to explore the role of filial piety in the suicidal behavior of Chinese women. Qualitative interviews were conducted with Chinese women with a history of suicidal behavior living in the Beijing area (n = 29). Filial piety data were extracted and analyzed in accordance with constructivist grounded theory. The women described five specific family and filial piety factors and how they influenced their ability to fulfill family role obligations, which was described as a nexus connecting these factors to depression, suicidal behavior, and recovery. The five factors were: 1) rigidity of parental filial expectations, 2) perception of family relationships as positive/supportive or negative/harsh, 3) whether filial piety is of high or low personal value in the woman's life, 4) any experiences of rebellion leading to punitive consequences, and 5) how much filial piety she receives from her children. These factors could inform suicide risk assessments in this population. They can be harnessed as part of recovery and protect against future suicidal behavior.


Alcohol's harms to others: the self-rated health of those with a heavy drinker in their lives.
Studies have examined the self-rated health (SRH) of the drinker, but only few have examined the health of those affected by a heavy drinker. This Nordic study aimed to examine the association between exposure to heavy drinkers and SRH. Data come from surveys from the five Nordic countries that participated in the Reducing Alcohol-Related Harm Standardized European Survey in 2015 (n = 7065 aged 18-64 years). Variables included a five-point Likert-scale question on one's SRH, a question on whether the respondent knew a heavy drinker in the last 12 months, and covariates. The 'fair', 'poor' and 'very poor' response categories were combined and are referred to as poor SRH. Multiple logistic regression models were used to examine the relationship between knowing a heavy drinker and one's SRH. Country-pooled adjusted analyses showed a significant relationship between knowing (and being negatively affected by) a heavy drinker and poor SRH [odds ratios (OR) = 1.39, 95% confidence intervals (CI): 1.02-1.89 for heavy drinker in household; OR = 1.23, 95% CI: 1.07-1.42 for other known heavy drinker, compared to not knowing a heavy drinker or knowing a heavy drinker, but not being negatively affected]. A graded relationship appeared such that increasing proximity of the known heavy drinker increased likelihood to report poor SRH. Knowing and being negatively affected by someone close who drinks heavily increases the likelihood of reporting poor SRH. These results have implications for public health messaging regarding the well-being of relatives of heavy drinkers.


Insulin resistance is associated with the aggressiveness of pancreatic ductal carcinoma.
To study whether insulin resistance accelerates the development and/or the progression of pancreatic adenocarcinoma (PDAC), we hypothesized that patients with insulin resistance, compared with those without insulin resistance, show: (1) a younger age and more advanced PDAC stage at diagnosis and (2) a shorter disease-free and overall survival after PDAC diagnosis. Prospective observational study of patients admitted to a referral center for pancreatic disease. Insulin resistance was defined as a HOMA-IR value greater than the 66th percentile value of the patients included in this study. Survival was estimated according to Kaplan-Meier and by Cox regression. Of 296 patients with PDAC, 99 (33 %) met criteria for being classified as insulin resistant at diagnosis. Median follow-up time after diagnosis was 5.27 ± 0.23 years. Patients with insulin resistance received a diagnosis of PDAC at a similar age compared to patients without insulin resistance (67.1 ± 9 vs. 66.8 ± 10 years, p = 0.68), but were more likely to have a cancer stage ≥3 (23.2 vs. 14.2 %, p = 0.053) and a residual disease after surgery (R1 56.4 vs. 38 %; p = 0.007). The median overall survival was 1.3 ± 0.14 and 1.79 ± 0.11 years for the patients with and without insulin resistance, respectively (p = 0.016). Results did not change when patients with diabetes at PDAC diagnosis were excluded from the analysis. Multivariate analysis showed that insulin resistance was independently associated with overall survival. Insulin resistance is associated with the aggressiveness of PDAC.


Lipoatrophic panniculitis: a possible autoimmune inflammatory disease of fat. Report of three cases.
We observed three children with a clinically similar presentation of erythematous nodules that expanded centrifugally leaving lipoatrophy. Areas of lipoatrophy coalesced, resulting in clinical pictures similar to partial or total lipodystrophy. Histologic study revealed a lobular panniculitis with a mixed infiltrate of lymphocytes and mononuclear phagocytes. Of these three children, one had insulin-dependent diabetes mellitus and Hashimoto's thyroiditis, one developed juvenile rheumatoid arthritis, and the third developed insulin-dependent diabetes mellitus, suggesting that the pathogenic mechanism may be an expression of autoimmunity.


Spanking and adult mental health impairment: The case for the designation of spanking as an adverse childhood experience.
Adverse Childhood Experiences (ACEs) such as child abuse are related to poor health outcomes. Spanking has indicated a similar association with health outcomes, but to date has not been considered an ACE. Physical and emotional abuse have been shown in previous research to correlate highly and may be similar in nature to spanking. To determine if spanking should be considered an ACE, this study aimed to examine 1): the grouping of spanking with physical and emotional abuse; and 2) if spanking has similar associations with poor adult health problems and accounts for additional model variance. Adult mental health problems included depressive affect, suicide attempts, moderate to heavy drinking, and street drug use. Data were from the CDC-Kaiser ACE study (N=8316, response rate=65%). Spanking loaded on the same factor as the physical and emotional abuse items. Additionally, spanking was associated with increased odds of suicide attempts (Adjusted Odds Ratios (AOR)=1.37; 95% CI=1.02 to1.86), moderate to heavy drinking (AOR)=1.23; 95% CI=1.07 to 1.41), and the use of street drugs (AOR)=1.32; 95% CI=1.4 to 1.52) in adulthood over and above experiencing physical and emotional abuse. This indicates spanking accounts for additional model variance and improves our understanding of these outcomes. Thus, spanking is empirically similar to physical and emotional abuse and including spanking with abuse adds to our understanding of these mental health problems. Spanking should also be considered an ACE and addressed in efforts to prevent violence.


[BRAF mutation testing for the choice of melanoma treatment].
Individual life-time risk of melanoma in white residents of highly developed countries may be as high as 2%. Continuing rise of melanoma incidence is directly related to the improving life standards, especially to growing opportunities of attending sea resorts and getting tanned. Melanoma is usually highly aggressive and resistant to standard cytotoxic therapy, therefore 5-years survival of patients with the metastatic disease does not exceed 10-15%. Approximately 50% of melanomas contain point mutation in codon 600 of the BRAF kinase. Specific inhibitors of activated BRAF have demonstrated unprecedented therapeutic efficacy, thus BRAF testing has become a mandatory component of treatment planning for inoperable melanoma. This review discusses key issues, which are related to various clinical, morphological, and molecular genetic aspects of determination of BRAF status in metastatic melanoma.


Iterative organometallic addition to chiral hydroxylated cyclic nitrones: highly stereoselective syntheses of alpha,alpha'- and alpha,alpha-substituted hydroxypyrrolidines.
[reaction: see text]. Iteration of organometallic addition to chiral hydroxylated pyrroline N-oxides through an addition-oxidation-addition synthetic sequence allowed highly stereoselective double alkylation of pyrrolidine at C-2 or at C-2 and C-5 depending on the regioselectivity of the oxidation step. Application of this methodology has been exemplified by the synthesis of the all-substituted pyrrolidine alkaloid (-)-codonopsinine and of proline-type amino acid precursors possessing a quaternary stereogenic center, whose configuration can be controlled.


[Health damage caused by smoking].
This paper describes the detrimental effects of smoking. More than 30% of the male and female Dutch population still smokes cigarettes. This figure compares unfavorably in international perspective. The smoking habit is responsible for approximately one third of all deaths from cardiovascular, pulmonary and malignant disease. Cigarette smokers face a 50% chance of dying as a result of a smoking related disease. The morbidity associated with smoking is also very substantial. Subjects who discontinue smoking before being struck by disease, and patients who quit the habit once early symptoms develop, have a more favorable outcome. For these reasons, and also because most patients esteem the opinion of their doctor, smoking behavior should be discussed during regular patient doctor contacts, and followed by the advice to quit smoking.


[Endomyocardial biopsy as a method of surveilling adriamycin cardiac toxicity. 28 biopsies in 26 patients].
The aim of this study was to evaluate the role of endomyocardial biopsy in the surveillance of toxic, latent cardiomyopathy secondary to adriamycin (ADM) therapy, and to assess its value by comparison with non-invasive investigations such as echocardiography. Twenty six patients underwent endomyocardial biopsy and echocardiographic measurement of the fractional shortening of the left ventricular internal diameter after a total dose of 150 to 520 mg/m2 of ADM. Interstitial fibrosis was observed in 8 of the 20 interpretable cases on light microscopy (6 moderate, 2 severe). There were no cellular changes. Slight focal myofibril degeneration was observed in 8 cases, and vacuolisation corresponding to mild dilatation of the sarcoplasmic reticulum was observed in 9 cases out of the 22 technically interpretable cases on electron microscopy. In contrast to the findings of interstitial fibrosis, cellular degeneration correlated significantly with the total dose of ADM (p less than 0,01). We conclude that this histological study confirms the fact that myocardial changes are rare and slight with doses of less than 520 mg/m2 of ADM and that non-invasive techniques are adequate for follow-up until this dose is reached.


Prevention and Management of Urgent/Emergent Limb Ischemia.
Patients who require urgent or emergent peripheral revascularization represent one of the highest risk subgroups of PAD patients. They suffer unacceptably high complication rates including recurrent ALI, vascular amputation, and death. In this article, we examine (1) the burden of cardiovascular complications according to PAD severity, (2) discuss medical optimization to improve vascular outcomes in symptomatic LE-PAD patients, and (3) review the evidence for management of patients following urgent/emergent limb ischemia. The VOYAGER trial recently demonstrated that rivaroxaban 2.5 mg BID + ASA daily significantly reduces major adverse cardiac and limb events in patients following lower extremity revascularization. A recent Canadian survey also demonstrated that significant heterogeneity exists in antithrombotic prescribing practices following urgent/emergent revascularization. COMPASS and VOYAGER have demonstrated the efficacy of aspirin 81 mg daily and rivaroxaban 2.5 mg twice daily at reducing MACE and MALE events in stable PAD patients and those undergoing elective revascularization. Patients who require urgent or emergent peripheral revascularization remain the highest thrombotic risk subgroup of PAD patients, in whom there is insufficient evidence to guide antithrombotic therapy. Despite clear evidence that multi-modal medical therapy (including statins, antihypertensive agents and smoking cessation) benefits patients with atherosclerosis, their use remains unacceptably low in PAD, and greater efforts are needed to understand and address patient, health provider, and system issues that prevent their optimal implementation in practice.


Bifrontal transcranial direct current stimulation modulates fatigue in multiple sclerosis: a randomized sham-controlled study.
Fatigue is a frequent and debilitating symptom in patients with central nervous system diseases. Up to 90% of patients with multiple sclerosis (MS) suffer from fatigue that drastically affects the quality of life. MS patients also complain of anxiety and depressive symptoms and these three manifestations tend to cluster together in this clinical population. The objective of this work was to assess the effects of transcranial direct stimulation (tDCS), a noninvasive brain stimulation technique, on fatigue as well as anxiety and depressive symptoms. Eleven fatigued MS patients randomly received two blocks (active and sham tDCS) of five consecutive daily sessions of bifrontal tDCS (anode/cathode over the left/right prefrontal cortices, respectively) in a crossover manner, separated by a 3-week washout interval. Evaluation took place at day 1, day 5 (right after each block) and 1 week later. Active but not sham tDCS resulted in a significant improvement of fatigue at day 5 (p < 0.05), an effect that seems to last at least 1 week following the stimulation (p = 0.05). Active tDCS also significantly improved anxiety symptoms, but the effect emerged 1 week later (p < 0.05). No significant effects were obtained regarding depression (p > 0.05). Bifrontal tDCS seems to modulate fatigue in PwMS. The observed anxiolytic effects could constitute delayed after effects of tDCS or might be mediated by fatigue improvement. These findings merit to be addressed in large-scale controlled trials.


Toxigenic fungi in food.
Forty-five fungal isolates from moldy supermarket foods were tested for toxicity to brine shrimp, and twenty-two of these isolates were subsequently tested for toxicity to chicken embryos. Highly toxigenic fungi were Cladosporium sphaerospermum from a bakery product, Fusarium oxysporum from carrots, F. solani from cabbage, Aspergillus niger and Penicillium corylophilum from bread, P. cyclopium and P. herguei from corn meal, P. lanosum from onions,P. steckii from chocolate syrup, Penicillium sp. from jelly, and Rhizopus nigricans isolates from sweet potato, applesauce, and strawberries. Approximately one-third of the fungal cultures were moderately to highly toxigenic to brine shrimp and chicken embryos, while several additional cultures were slightly toxigenic.


Predicting binding affinity of CSAR ligands using both structure-based and ligand-based approaches.
We report on the prediction accuracy of ligand-based (2D QSAR) and structure-based (MedusaDock) methods used both independently and in consensus for ranking the congeneric series of ligands binding to three protein targets (UK, ERK2, and CHK1) from the CSAR 2011 benchmark exercise. An ensemble of predictive QSAR models was developed using known binders of these three targets extracted from the publicly available ChEMBL database. Selected models were used to predict the binding affinity of CSAR compounds toward the corresponding targets and rank them accordingly; the overall ranking accuracy evaluated by Spearman correlation was as high as 0.78 for UK, 0.60 for ERK2, and 0.56 for CHK1, placing our predictions in the top 10% among all the participants. In parallel, MedusaDock, designed to predict reliable docking poses, was also used for ranking the CSAR ligands according to their docking scores; the resulting accuracy (Spearman correlation) for UK, ERK2, and CHK1 were 0.76, 0.31, and 0.26, respectively. In addition, performance of several consensus approaches combining MedusaDock- and QSAR-predicted ranks altogether has been explored; the best approach yielded Spearman correlation coefficients for UK, ERK2, and CHK1 of 0.82, 0.50, and 0.45, respectively. This study shows that (i) externally validated 2D QSAR models were capable of ranking CSAR ligands at least as accurately as more computationally intensive structure-based approaches used both by us and by other groups and (ii) ligand-based QSAR models can complement structure-based approaches by boosting the prediction performances when used in consensus.


Impact of the COVID-19 and Socioeconomic Status on Access to Care for Otorhinolaryngology Patients.
Since December 2019, COVID-19 has greatly influenced public healthcare systems around the globe in various aspects, including limitation of healthcare accessibility due to lack of both human and financial resources, suspension of clinics, and fear of infection causing healthcare avoidance. The aim of this study was to investigate the impact of COVID-19 on access to healthcare for otorhinolaryngology patients from different socioeconomic status (SES) groups. Otorhinolaryngology patients' disease severity status, diagnosed at the first hospital visit, was investigated during the pre -and post-COVID-19 pandemic era in a single medical center located in Seoul, Korea. An ordinal regression model was used to assess the impact of both SES and the COVID-19 pandemic on otorhinolaryngology diseases. Within the chronic rhinosinusitis group, lower SES was associated with a higher disease severity at the first visit compared to higher SES (OR = 3.25). During the COVID-19 pandemic, while the total number of outpatients was reduced, the severity of these ENT diseases seemed to increase compared to the pre-pandemic severity in every SES group. Our study demonstrates the negative impact a worldwide pandemic can have on healthcare inequity and disease severity, and highlights the importance of re-allocating fundamental resources for those in need during periods of public health crisis.


The Journal of Human Lactation: A Reflective Discussion.
Research about lactation and breastfeeding has exploded since the Journal of Human Lactation (JHL) began publishing in 1985. To discuss the 3-decade-long role of the Journal in promoting, supporting, and disseminating lactation research, editors convened a multidisciplinary group of lactation researchers and providers which comprised three senior researchers and one clinical practitioner, all of whom have a long history of lactation advocacy. Their discussion took place on July 1, 2019. Dr Aimee Eden is a medical anthropologist who leads the qualitative research efforts in a small research department. Her dissertation research focused on the maternal and child healthcare workforce, and the professionalization of breastfeeding support. She served on the Board of Directors of the International Board of Lactation Examiners (2010-2016) and currently serves on the board of the Monetary Investment for Lactation Consultant Certification. Dr Karleen Gribble has been conducting research for 15 years about infant and young child feeding in emergencies, long-term breastfeeding, milk sharing, early childhood trauma, adoption, and fostering. She is an Australian Breastfeeding Association community educator and breastfeeding counselor and a member of the Infant and Young Child Feeding in Emergencies Core Group. Elien Rouw is a practicing physician in Germany specialized in healthy infant care, with a long-standing specialization in breastfeeding medicine. She serves on the Board of Directors of the Academy of Breastfeeding Medicine, is a member of the German National Breastfeeding Committee and their delegate to the World Alliance for Breastfeeding Action. Dr Jackie Wolf is an historian of medicine, whose research focuses on the history of childbirth and breastfeeding practices in the United States and how those practices have shaped women's and children's health, as well as public health, over time. Her latest book, published by Johns Hopkins University Press and funded by a 3 year grant from the National Institutes of Health, is Cesarean Section: An American History of Risk, Technology, and Consequence. Dr Sara Gill moderated the discussion. She was a member of the Board of Directors of the International Lactation Consultant Association for 5 years, and has been an Associate Editor of the Journal of Human Lactation for the past 4 years. Her research has focused on breastfeeding among vulnerable populations. (Participants' comments are noted as AE = Aimee Eden; SG = Sara Gill; KG = Karleen Gribble; ER = Elien Rouw; JW = Jacqueline Wolf).


Prevalence and Surgical Outcomes of Macular Hole in Eyes with Age-Related Macular Degeneration.
To report the prevalence and surgical outcomes of macular holes (MHs) in eyes with age-related macular degeneration (AMD). Interventional, retrospective, consecutive case series. Patients with MH and concurrent non-neovascular (NNV) or neovascular (NV) AMD. The records of 27 912 patients diagnosed with AMD between 2009 and 2014 at Associated Retinal Consultants were reviewed. Demographic data, visual acuity (VA), funduscopic examination, and optical coherence tomography were reviewed in those with a concurrent diagnosis of MH. The VA and MH closure status. A total of 15 196 patients with NNV and 12 716 patients with NV AMD were identified. A total of 199 eyes (0.7%) had MHs (160 NNV [1.1%]; 39 NV [0.3%]). Mean time to diagnosis of MH after the initial visit was 11.2 months (7.1 NNV; 24.8 NV). A total of 127 eyes underwent surgical repair (106 NNV; 21 NV). The final closure rate in those who underwent vitrectomy was 89.8% (91.5% NNV; 81.0% NV) and 25.0% in those who were observed (18.5% NNV, P < 0.0001; 44.4% NV, P = 0.02). Preoperative logarithm of the minimum angle of resolution VAs in NNV and NV AMD was 0.8±0.4 and 0.8±0.5, respectively, and final VA was 0.6±0.5 (P < 0.001) and 0.9±0.6 (P = 0.52), respectively. Mean follow-up time was 5.0 years. The prevalence of MH was higher in eyes with NNV AMD than in those with NV AMD. The surgical closure rate was comparable in both groups, but VA improvement reached statistical significance only in the NNV AMD group.


Differences in host species relationships and biogeographic influences produce contrasting patterns of prevalence, community composition and genetic structure in two genera of avian malaria parasites in southern Melanesia.
Host-parasite interactions have the potential to influence broadscale ecological and evolutionary processes, levels of endemism, divergence patterns and distributions in host populations. Understanding the mechanisms involved requires identification of the factors that shape parasite distribution and prevalence. A lack of comparative information on community-level host-parasite associations limits our understanding of the role of parasites in host population divergence processes. Avian malaria (haemosporidian) parasites in bird communities offer a tractable model system to examine the potential for pathogens to influence evolutionary processes in natural host populations. Using cytochrome b variation, we characterized phylogenetic diversity and prevalence of two genera of avian haemosporidian parasites, Plasmodium and Haemoproteus, and analysed biogeographic patterns of lineages across islands and avian hosts, in southern Melanesian bird communities to identify factors that explain patterns of infection. Plasmodium spp. displayed isolation-by-distance effects, a significant amount of genetic variation distributed among islands but insignificant amounts among host species and families, and strong local island effects with respect to prevalence. Haemoproteus spp. did not display isolation-by-distance patterns, showed marked structuring of genetic variation among avian host species and families, and significant host species prevalence patterns. These differences suggest that Plasmodium spp. infection patterns were shaped by geography and the abiotic environment, whereas Haemoproteus spp. infection patterns were shaped predominantly by host associations. Heterogeneity in the complement and prevalence of parasite lineages infecting local bird communities likely exposes host species to a mosaic of spatially divergent disease selection pressures across their naturally fragmented distributions in southern Melanesia. Host associations for Haemoproteus spp. indicate a capacity for the formation of locally co-adapted host-parasite relationships, a feature that may limit intraspecific gene flow or range expansions of closely related host species.


The effect of associative strength on semantic priming in schizophrenia.
The present research was designed to investigate the pattern of semantic priming in schizophrenia as a function of strength of association (or semantic distance between concepts in the semantic network). Thirty schizophrenia patients, without formal thought disorder, and twenty-nine healthy controls participated in a lexical decision task in which prime-target associative strength (strong, weak and not related) and stimulus-onset asynchrony (SOA: 250ms and 750ms) were manipulated. Patients and controls showed the same associative strength effect on RTs. In the short SOA condition priming effects were obtained for both strong and weak prime-target associative conditions. However in the long SOA priming was only significant for strongly associated pairs. This pattern of priming effects was similar in both groups, with higher priming on the short SOA and strong association conditions. Altogether results suggest that automatic semantic spreading activation is unimpaired in schizophrenia patients without formal thought disorder. These results are in line with the general evidence of impaired implicit priming observed only in patients with formal thought disorder. At the same time patients use context as controls to facilitate word processing. Finally, these findings evidence that, prime-target associative strength could moderate results in studies of semantic memory deficits in schizophrenia.


Localization of the high-affinity glutamate transporter EAAC1 in rat kidney.
Most amino acids filtered by the glomerulus are reabsorbed in the kidney via specialized transport systems. Recently, the cDNA encoding a high-affinity glutamate transporter, EAAC1, has been isolated and shown to be expressed at high levels in the kidney. To determine the potential role of EAAC1 in renal acidic amino acid reabsorption, the distribution of EAAC1 mRNA and protein in rat kidney was examined. In situ hybridization revealed that EAAC1 mRNA is expressed predominantly in S2 and S3 segments of the proximal tubules and at low levels in the inner stripe of outer medulla and inner medulla. Polyclonal antibodies raised against the carboxy terminus of EAAC1 recognized a single band of approximately 70 kDa on Western blots of membrane protein from kidney cortex and medulla. Immunofluorescence microscopy revealed intense signals in the luminal membrane of S2 and S3 segments and weaker signals in S1 segments, descending thin limbs of long-loop nephrons, medullary thick ascending limbs, and distal convoluted tubules. These results are consistent with EAAC1 encoding the previously described apical high-affinity glutamate transporter in the kidney that mediates reabsorption of acidic amino acids in tubules beyond early proximal tubule S1 segments. Potential additional roles of EAAC1 in acid/base balance, cell volume regulation, and amino acid metabolism are discussed.


Analyzing Medical Research Results Based on Synthetic Data and Their Relation to Real Data Results: Systematic Comparison From Five Observational Studies.
Privacy restrictions limit access to protected patient-derived health information for research purposes. Consequently, data anonymization is required to allow researchers data access for initial analysis before granting institutional review board approval. A system installed and activated at our institution enables synthetic data generation that mimics data from real electronic medical records, wherein only fictitious patients are listed. This paper aimed to validate the results obtained when analyzing synthetic structured data for medical research. A comprehensive validation process concerning meaningful clinical questions and various types of data was conducted to assess the accuracy and precision of statistical estimates derived from synthetic patient data. A cross-hospital project was conducted to validate results obtained from synthetic data produced for five contemporary studies on various topics. For each study, results derived from synthetic data were compared with those based on real data. In addition, repeatedly generated synthetic datasets were used to estimate the bias and stability of results obtained from synthetic data. This study demonstrated that results derived from synthetic data were predictive of results from real data. When the number of patients was large relative to the number of variables used, highly accurate and strongly consistent results were observed between synthetic and real data. For studies based on smaller populations that accounted for confounders and modifiers by multivariate models, predictions were of moderate accuracy, yet clear trends were correctly observed. The use of synthetic structured data provides a close estimate to real data results and is thus a powerful tool in shaping research hypotheses and accessing estimated analyses, without risking patient privacy. Synthetic data enable broad access to data (eg, for out-of-organization researchers), and rapid, safe, and repeatable analysis of data in hospitals or other health organizations where patient privacy is a primary value.


Magnetic resonance imaging of pediatric hip disease.
The efficacy of magnetic resonance imaging (MRI) in the assessment of pediatric hip disease was tested by scanning the hips of 24 children (30 scans). Twelve patients with Legg-Calvé-Perthes disease (17 hips) showed characteristic areas of low-intensity signal representative of necrotic areas of the capital epiphysis. Abnormal scans were also obtained on patients with transient synovitis, avascular necrosis secondary to steroids, epiphyseal dysplasia, and multiple osteochondromatosis. MRI accurately shows articular cartilage, femoral head shape, quality of containment, and areas of necrosis in pediatric hips. The extent of involvement and revascularization can be identified in Legg-Calvé-Perthes disease.


Characterization and differential regulation of GABAA and benzodiazepine receptors in rat neocortex.
We have characterized the gamma-aminobutyric acid-A (GABAA) and benzodiazepine (BZ) receptors in in vitro living slices of adult rat neocortex using [3H]SR95531, a GABAA antagonist, and [3H]flunitrazepam (FNZ), a BZ ligand. [3H]SR95531 labelled a single population of GABAA receptors with a Bmax of 1030.7 fmol/mg protein and a Kd of 43.5 nM. [3H]FNZ also labelled a single binding site with a Bmax of 4239 fmol/mg protein and a Kd of 22 nM. The GABAA receptor labelled using [3H]SR95531 could be down-regulated by 2 h preincubations in GABA and the GABAA agonist muscimol (8% and 11%, respectively). Increases in cellular electrical activity induced by a combination of veratridine and glutamate led to an average increase in GABAA receptor number of 58%. The BZ binding site labelled with [3H]FNZ was down-regulated by clonazepam (-55%), increased by GABA (+17%), but not altered by changes in electrical activity. The present results demonstrate the rapid differential regulation of a ligand-gated receptor by agonist stimulation or increases in bioelectric activity. Such regulation may provide clues to the nature of the modifications which occur following changes in cellular activity in the cortex.


[Current data on iron metabolism].
Iron is required for cellular life. However, abnormalities of its metabolism may lead to iron deficiency or iron overload, both conditions which are deleterious. Therefore, stock and distribution of iron in the body must be very stable. Classically, four major proteins are involved in iron metabolism: (a) transferrin which is implicated in its plasmatic transport, (b) transferrin receptor which regulates iron-transferrin uptake, (c) ferritin, the major iron storage protein, and (d) IRP (Iron Regulatory Protein) which regulates both the entry and storage of iron by linking to the IRE (Iron Responsive Element), a nucleotidic sequence found on transferrin receptor and ferritin mRNA. Thus, IRP adapts gene expression to the iron cellular status. Recent data give informations about new proteins involved in iron metabolism: HFE whose gene is mutated in genetic hemochromatosis, ceruloplasmin which permits cellular iron egress and frataxin which is implicated in the exit of iron from mitochondria.


CYP2E1 and clinical features in alcoholics.
It is known that P-4502E1 metabolizes both ethanol and acetaldehyde, and that it can be induced after birth by inducers such as ethanol. Genetic polymorphisms in the 5'-flanking region change its transcriptional regulation. Persons who are either hetereozygous or mutant homozygous for CYP2E1 (C1/C2 or C2/C2) can drink much more alcohol than persons who are normal homozygous for CYP2E1 (C1/C1) among those whose aldehyde dehydrogenase 2 genotype is heterozygous. Subjects were composed of 52 Japanese alcoholics. The alcoholics having no C2 (group 1) and the alcoholics having C2 (group 2) were compared with each other with respect to their clinical characteristics: family history, past history, alcohol-related symptoms, complications, social disabilities, self-consciousness and short alcohol dependence date. In the social disabilities, the genotypes of CYP2E1 (C1/C2 or C2/C2) showed significantly more cases of guilts (p < 0.05). In regard to liver disease or self-consciousness, this was at a trend level (p = 0.08). We have shown that the genotypes of CYP2E1 are associated with clinical features of alcoholics.


Exploring the factors contributing to sibling correlations in BMI: a study using the Panel Study of Income Dynamics.
Understanding the mechanisms contributing to correlated BMI outcomes in a social network such as siblings will help policy makers reduce the burden of disease associated with obesity. There are two potential mechanisms explaining correlated BMI outcomes in a biologically related social network: (i) time constant factors such as genetic heritability and habits formed during childhood and (ii) factors that change over time some of which are dependent on the frequency of interactions between the social network, for example, social norms shaped by the social network's shifting attitudes towards weight and behaviors related to weight, or environmental factors like opportunities for exercise. This study aims to distinguish between time constant factors from factors that are likely to change over time to gain a better understanding of the mechanisms explaining the correlation in sibling BMI. We exploit data from the Panel Study of Income Dynamics (PSID) over 1999-2007 estimating the correlation in BMI for adult siblings who currently live in separate households but grew-up in the same household and adolescent siblings currently living in the same household to isolate the influence of factors that change over time. The findings indicate that time constant factors explain some of the overall correlation in sibling BMI for both cohorts of siblings. Factors that change over time only significantly impact on the overall correlation in BMI for adolescent siblings suggesting if there is a social network influence on correlations in BMI this is facilitated by sharing the same household.


Efficacy of single-agent immunosuppressive regimens in a murine model of vascularized composite allotransplantation.
We herein investigate the safety and efficacy of single-agent anti-rejection regimens in a mouse vascularized composite allotransplantation (VCA) model. Orthotopic hind-limb transplantations (Balb/c → C57BL/6) were performed using 6- to 8-week-old mice. A thirty-day regimen of either rapamycin, tacrolimus (both 1, 3, 5 mg/kg/day) or cyclosporine (25, 35, 50 mg/kg/day) was used. Primary endpoints were animal and graft survival, and secondary chimerism and regulatory T-cell levels. For rapamycin and tacrolimus given at 1, 3, and 5 mg/kg/day, median graft survival time (MST) was 23 days (18-28 days), 30 days (23-30 days), and 30 d (30-30 days) and 14 days (13-18 days), 30 days (16-30 days), and 30 days (30-30 days), respectively. For cyclosporine dosed at 25 and 35 mg/kg/day, MST was 15 days (12-18 days) and 21 days (14-27 days). Toxicity from CsA 50 mg/kg led to 100% mortality. Mixed chimerism levels were higher in rapamycin-treated animals than in tacrolimus-treated recipients (P = 0.029). Tacrolimus was superior in preventing leukocyte recruitment to the allograft. In murine VCA, no standardized immunosuppressive regimen exists, limiting comparability of outcomes and survival. Our data demonstrate that rapamycin and tacrolimus maintenance treatment at 5 mg/kg/day both yielded allograft survival (<grade 3 rejection) in all animals. Rapamycin displayed less toxicity and maintained mixed chimerism but was not as potent in controlling leukocyte recruitment compared with tacrolimus.


Predictors of prolonged hospital stay following open and laparoscopic gastric bypass for morbid obesity: body mass index, length of surgery, sleep apnea, asthma, and the metabolic syndrome.
The number of weight reduction operations performed for type II and type III obesity is rapidly escalating. Risk of surgery has been infrequently stratified for patient subgroups. The purpose of this study was to identify patient characteristics that increased the odds of a prolonged hospital length of stay (LOS) following open or laparoscopic Roux-en-Y gastric bypass (RYGBP). The hospital records of 311 patients who underwent RYGBP in a 6-month period were retrospectively reviewed. Patient characteristics including the presence of significant obesity-related medical conditions were recorded. Analysis was based on intent to treat. Univariate and step-wise logistic regression analysis was used to identify the odds ratio (OR) and adjusted odds ratio (AOR) for predictors of an increased hospital LOS. Datasets for 311 patients were complete.159 patients underwent open vertical banded gastro-plasty-Roux-en-Y gastric bypass (VBG-RYGBP) and152 laparoscopic RYGBP (LRYGBP). 78% of patients were female. Median age was 40 years (range 18-68). Median BMI was 49 kg/m2 (range 35-82). 17% of patients had sleep apnea, 18% asthma, 19% type 2 diabetes, 13% hypercholesterolemia and 44% hypertension. Median length of surgery for open VBG-RYGBP (64 minutes) was significantly faster than forLRYGBP (105 minutes). Median length of stay was significantly shorter for LRYGBP (2 days) than openVBG-RYGBP (3 days). Univariate logistic regression analysis identified 6 predictors of increased LOS:open surgery (0.4 OR); increasing BMI (60 kg/m2 0.38 OR; BMI 70 kg/m2 0.53 OR); increasing length of surgery (120 min 0.33 OR; 180 min 0.48 OR); sleep apnea (2.25 OR); asthma (3.73 OR); and hypercholesterolemia (3.73 OR). Subset analysis identified patients with the greatest odds for a prolonged hospital stay: women with asthma (2.47 AOR) or coronary artery disease (8.65 AOR); men with sleep apnea (5.54 OR) or the metabolic syndrome (6.67-10.20 OR); and patients undergoing a laparoscopic operation with sleep apnea (11.53 AOR) or coronary artery disease(12.15 AOR). Open surgery, BMI, length of surgery,sleep apnea, asthma and hypercholesterolemia all increased the odds of a prolonged LOS. Patients with the greatest odds of long LOS were women with asthma or coronary disease, men with sleep apnea or the metabolic syndrome, and patients undergoing laparoscopic surgery with sleep apnea or coronary artery disease. Patients at high-risk for prolonged hospital stay can be identified before undergoingRYGBP. Surgeons may wish to avoid high-risk patients early in their bariatric surgery experience.


Cryptic genetic gluten intolerance revealed by intestinal antitransglutaminase antibodies and response to gluten-free diet.
Antitransglutaminase (anti-TG2) antibodies are synthesised in the intestine and their presence seems predictive of future coeliac disease (CD). This study investigates whether mucosal antibodies represent an early stage of gluten intolerance even in the absence of intestinal damage and serum anti-TG2 antibodies. This study investigated 22 relatives of patients with CD genetically predisposed to gluten intolerance but negative for both serum anti-TG2 antibodies and intestinal abnormalities. Fifteen subjects were symptomatic and seven were asymptomatic. The presence of immunoglobulin A anti-TG2 antibodies in the intestine was studied by creating phage-antibody libraries against TG-2. The presence of intestinal anti-TG2 antibodies was compared with the serum concentration of the intestinal fatty acid-binding protein (I-FABP), a marker for early intestinal mucosal damage. The effects of a 12-month gluten-free diet on anti-TG2 antibody production and the subjects' clinical condition was monitored. Twelve subjects entered the study as controls. The intestinal mucosa appeared normal in 18/22; 4 had a slight increase in intraepithelial lymphocytes. Mucosal anti-TG2 antibodies were isolated in 15/22 subjects (68%); in particular symptomatic subjects were positive in 13/15 cases and asymptomatic subjects in 2/7 cases (p=0.01). No mucosal antibodies were selected from the controls' biopsies. There was significant correlation between the presence of intestinal anti-TG2 antibodies and positive concentrations of I-FABP (p=0.0008). After a gluten-free diet, 19/22 subjects underwent a second intestinal biopsy, which showed that anti-TG2 antibodies had disappeared in 12/15 (p=0.002), while I-FABP decreased significantly (p<0.0001). The diet resolved both extraintestinal and intestinal symptoms. A new form of genetic-dependent gluten intolerance has been described in which none of the usual diagnostic markers is present. Symptoms and intestinal anti-TG2 antibodies respond to a gluten free-diet. The detection of intestinal anti-TG2 antibodies by the phage-antibody libraries has an important diagnostic and therapeutic impact for the subjects with gluten-dependent intestinal or extraintestinal symptoms. Clinical trial number NCT00677495.


Effect of platelet-rich plasma with self-assembled peptide on the rotator cuff tear model in rat.
Several trials have been carried out to improve the healing of rotator cuff tear, but their effects remain controversial. In this study, we examined the therapeutic effect of platelet-rich plasma (PRP) in combination with self-assembled peptide (SAP) on the healing of rotator cuff tear in the rat. Twenty-seven Sprague-Dawley rats, aged 15 weeks, were used for the rotator cuff tear model. The supraspinatus muscle at the insertion site was dissected and truncated, leaving the cut edge free. A week after the surgery, the rats were randomly divided into SAP, PRP, SAP-PRP and control groups and 0.2 ml SAP, PRP, SAP-PRP and saline were injected, respectively. Bonar scores and synovial inflammation grade were checked 5 weeks after the injection. Immunofluorescence staining for heat shock protein (HSP)-70 and caspase-3 was conducted. Furthermore, maximum stride length was measured before and at 2 and 4 weeks after the injection. The Bonar scores were 6.4 ± 1.8 in the SAP group, 5.9 ± 2.0 in the PRP group, 4.7 ± 1.1 in the SAP-PRP group and 8.3 ± 2.3 in the control group. There was significant difference between the SAP-PRP and control groups in post hoc multiple comparison analysis. Among four categories of Bonar scores, collagen arrangement and vascular infiltration showed improvement after SAP-PRP injection. Immunofluorescence images showed that HSP-70 and caspase-3 were much less expressed in the SAP-PRP group than in the other groups. In conclusion, SAP-PRP can be effective in healing a rotator cuff tear by enhancing the collagen arrangement and inhibiting inflammatory changes and apoptosis. Copyright © 2015 John Wiley & Sons, Ltd.


Cysteine desulfurase (IscS)-mediated fine-tuning of bioenergetics and SUF expression prevents Mycobacterium tuberculosis hypervirulence.
Iron-sulfur (Fe-S) biogenesis requires multiprotein assembly systems, SUF and ISC, in most prokaryotes. M. tuberculosis (Mtb) encodes a complete SUF system, the depletion of which was bactericidal. The ISC operon is truncated to a single gene iscS (cysteine desulfurase), whose function remains uncertain. Here, we show that MtbΔiscS is bioenergetically deficient and hypersensitive to oxidative stress, antibiotics, and hypoxia. MtbΔiscS resisted killing by nitric oxide (NO). RNA sequencing indicates that IscS is important for expressing regulons of DosR and Fe-S-containing transcription factors, WhiB3 and SufR. Unlike wild-type Mtb, MtbΔiscS could not enter a stable persistent state, continued replicating in mice, and showed hypervirulence. The suf operon was overexpressed in MtbΔiscS during infection in a NO-dependent manner. Suppressing suf expression in MtbΔiscS either by CRISPR interference or upon infection in inducible NO-deficient mice arrests hypervirulence. Together, Mtb redesigned the ISC system to "fine-tune" the expression of SUF machinery for establishing persistence without causing detrimental disease in the host.


Reduced incidence of cardiac arrhythmias in walkers and runners.
Walking is purported to reduce the risk of atrial fibrillation by 48%, whereas jogging is purported to increase its risk by 53%, suggesting a strong anti-arrhythmic benefit of walking over running. The purpose of these analyses is to compare incident self-reported physician-diagnosed cardiac arrhythmia to baseline energy expenditure (metabolic equivalent hours per day, METhr/d) from walking, running and other exercise. Proportional hazards analysis of 14,734 walkers and 32,073 runners. There were 1,060 incident cardiac arrhythmias (412 walkers, 648 runners) during 6.2 years of follow-up. The risk for incident cardiac arrhythmias declined 4.4% per baseline METhr/d walked by the walkers, or running in the runners (P = 0.0001). Specifically, the risk declined 14.2% (hazard ratio: 0.858) for 1.8 to 3.6 METhr/d, 26.5% for 3.6 to 5.4 METhr/d, and 31.7% for ≥5.4 METhr/d, relative to <1.8 METhr/d. The risk reduction per METhr/d was significantly greater for walking than running (P<0.01), but only because walkers were at 34% greater risk than runners who fell below contemporary physical activity guideline recommendations; otherwise the walkers and runners had similar risks for cardiac arrhythmias. Cardiac arrhythmias were unrelated to walking and running intensity, and unrelated to marathon participation and performance. The risk for cardiac arrhythmias was similar in walkers and runners who expended comparable METhr/d during structured exercise. We found no significant risk increase for self-reported cardiac arrhythmias associated with running distance, exercise intensity, or marathon participation. Rhythm abnormalities were based on self-report, precluding definitive categorization of the nature of the rhythm disturbance. However, even if the runners' arrhythmias include sinus bradycardia due to running itself, there was no increase in arrhythmias with greater running distance.


Human chromosome banding with restriction endonucleases HaeIII, HinfI and PstI.
Human chromosome banding was carried out with restriction endonucleases (REs) HaeIII, HinfI, PstI and the effects of some factors on it were examined. HaeIII induced C and G banding patterns, HinfI induced negative C bands but the centromeric heterochromatin of chromosomes 3 and 4 remained selectively dark-stained. A new heteromorphism of C banded region was discovered in chromosome 4. PstI induced G-like banding pattern. The duration of enzymatic digestion, the concentration of glycerol in the reaction buffer, as well as the ageing and heating of chromosome preparations all had a significant influence on the banding effects of the REs.


Vitamin D and Influenza-Prevention or Therapy?
Vitamin D generates many extraskeletal effects due to the vitamin D receptor (VDR) which is present in most tissues throughout the body. The possible role of vitamin D in infections is implied from its impact on the innate and adaptive immune responses. A significant effect is also the suppression of inflammatory processes. Because vitamin D could be acknowledged as a "seasonal stimulus", as defined by R. Edgar Hope-Simpson, it would be crucial to prove it from a potential easy and cheap prophylaxis or therapy support perspective as far as influenza infections are concerned. The survey of the literature data generates some controversies and doubts about the possible role of vitamin D in the prevention of influenza virus. The most important point is to realise that the broad spectrum of this vitamin's activity does not exclude such a possibility. According to most of the authors, more randomized controlled trials with effective, large populations are needed to explore the preventive effect of vitamin D supplementation on viral influenza infections.


Epizootic of parainfluenza-3 virus infection in gibbons.
A laboratory-housed breeding colony of white-handed gibbons (Hylobates lar) experienced an epizootic of upper respiratory tract disease characterized by lethargy, anorexia, coughing, and serous rhinorrhea. Signs were more severe in adults than in offspring, and all animals recovered without complications. Base-line, acute, and convalescent sera from the most severely affected gibbons were tested for antibodies against a wide spectrum of infectious agents. For personnel known to have had contact with the gibbons, testing for the same agents was done on base-line sera and sera obtained at the same time as the acute and convalescent sera were obtained from the gibbons. Rising titers against parainfluenza-3 virus were detected in 6 of 7 gibbons tested. An increase in titer was not seen in the sera of personnel.


[State of intracellular signaling systems in endothelial dysfunction].
To estimate the state of different parts of intracellular signaling system associated with NO production after a2-adrenoreceptors stimulation under endothelial dysfunction in 22 patients with diabetic foot, the inhibitory analysis of clophelin-induced platelets aggregation modulation by administration of eNOS stimulator (L-arginin) and inhibitor (L-NAME), modulator of Ca-calmoduline (triphtazine), guanilate cyclase stimulator (sodium nitroprusside), inhibitors of phosphodiesterases (theophillinum) and protein kinase A (butamide). It was shown that endothelial dysfunction in patients with diabetic foot is associated with alteration of adrenoreactivity and inhibition of eNOS, guanilate cyclase, protein kinase A and stimulation of phosphodiesterases activity.


Animals with a schizophrenia-like phenotype are differentially sensitive to the motivational effects of cannabinoid agonists in conditioned place preference.
Cannabis is the most consumed illicit drug worldwide, but among patients with a diagnosis of schizophrenia, this consumption is higher suggesting that they are differentially sensitive to cannabis. We chose to study this problematic using a neurodevelopmental model of schizophrenia: neonatal ventral hippocampus lesions (NVHL). In a first study, we compared the locomotor response to novelty, a mild stress and two doses of amphetamine (0.75 and 1.5 mg/kg) in sham and NVHL rats at post-natal day 35 (PD35) or 56 (PD56). In a second study, we investigated the valence of the motivational effect of Delta-9-tetrahydrocannabinnol (THC, 0.5 mg/kg, i.p.) and the cannabinoid receptor agonist, WIN55,212-2 (WIN, 1 mg/kg, i.p.), using the conditioned place preference paradigm; we used a biased procedure that comprised 12 days of testing with 3 paired-conditioning. The effects of this dose of WIN were also measured on locomotor activity. Results confirmed that the adult NVHL animals displayed a stronger locomotor response to the two doses of amphetamine, but not to novelty and a mild stress. In adult NVHL, but not sham animals, WIN stimulated locomotor activity and produced a conditioned place aversion. At the dose tested, THC tended to produce an aversion in adult sham but not NVHL animals. Taken together these findings show that adult animals with a schizophrenia-like phenotype are differentially sensitive to the motivational effect of cannabinoids.


Impact and cost-effectiveness of measles vaccination through microarray patches in 70 low-income and middle-income countries: mathematical modelling and early-stage economic evaluation.
Microarray patches (MAPs) are a promising technology being developed to reduce barriers to vaccine delivery based on needles and syringes (N&S). To address the evidence gap on the public health value of applying this potential technology to immunisation programmes, we evaluated the health impact on measles burden and cost-effectiveness of introducing measles-rubella MAPs (MR-MAPs) in 70 low-income and middle-income countries (LMICs). We used an age-structured dynamic model of measles transmission and vaccination to project measles cases, deaths and disability-adjusted life-years during 2030-2040. Compared with the baseline scenarios with continuing current N&S-based practice, we evaluated the introduction of MR-MAPs under different measles vaccine coverage projections and MR-MAP introduction strategies. Costs were calculated based on the ingredients approach, including direct cost of measles treatment, vaccine procurement and vaccine delivery. Model-based burden and cost estimates were derived for individual countries and country income groups. We compared the incremental cost-effectiveness ratios of introducing MR-MAPs to health opportunity costs. MR-MAP introduction could prevent 27%-37% of measles burden between 2030 and 2040 in 70 LMICs, compared with the N&S-only immunisation strategy. The largest health impact could be achieved under lower coverage projection and accelerated introduction strategy, with 39 million measles cases averted. Measles treatment cost is a key driver of the net cost of introduction. In countries with a relatively higher income, introducing MR-MAPs could be a cost-saving intervention due to reduced treatment costs. Compared with country-specific health opportunity costs, introducing MR-MAPs would be cost-effective in 16%-81% of LMICs, depending on the MR-MAPs procurement prices and vaccine coverage projections. Introducing MR-MAPs in LMICs can be a cost-effective strategy to revitalise measles immunisation programmes with stagnant uptake and reach undervaccinated children. Sustainable introduction and uptake of MR-MAPs has the potential to improve vaccine equity within and between countries and accelerate progress towards measles elimination.