The German version of the revised Adolescent Sleep-Wake Scale (rASWS) - A validation study in pediatric pain patients and school children.
Sleep problems are common in children and adolescents with chronic pain. The revised Adolescent Sleep-Wake Scale (rASWS) is an internationally well-established instrument to assess sleep quality in adolescents. So far, no German version is available. The study aimed to provide a validated German version of the rASWS, specifically for use in children and adolescents with chronic pain. The translated questionnaire was validated in a sample of N = 159 pediatric outpatients with chronic pain (8-17 years; 65.4% female), who presented to a specialized pediatric pain center. For cross-validation a community sample of N = 1348 school children was analyzed. Confirmatory factor analysis was conducted to examine the factor structure of the original 10-item 3-factor model in the sample of children and adolescents with chronic pain, which showed poor model fit. Model modifications were carried out by deleting 3 items with low factor loadings stepwise. The overall model fit of the final 3-factor model containing 7 items was excellent. Cronbach's α of the derived scales ranged from 0.74 to 0.86. Cross-validation in a community sample of school children confirmed the superiority of the 7-item model. The convergent validity of the measure was proved by moderate correlations between the rASWS and self-reported sleep problems. Associations with chronic pain characteristics were evident for pain-related disability. The use of the 7-item version of the rASWS for German-speaking children and adolescents with and without chronic pain is recommended as a self-report measure of sleep quality.


Long-term expression of human coagulation factor VIII and correction of hemophilia A after in vivo retroviral gene transfer in factor VIII-deficient mice.
Hemophilia A is caused by a deficiency in coagulation factor VIII (FVIII) and predisposes to spontaneous bleeding that can be life-threatening or lead to chronic disabilities. It is well suited for gene therapy because a moderate increase in plasma FVIII concentration has therapeutic effects. Improved retroviral vectors expressing high levels of human FVIII were pseudotyped with the vesicular stomatitis virus G glycoprotein, were concentrated to high-titers (10(9)-10(10) colony-forming units/ml), and were injected intravenously into newborn, FVIII-deficient mice. High-levels (>/=200 milliunits/ml) of functional human FVIII production could be detected in 6 of the 13 animals, 4 of which expressed physiologic or higher levels (500-12,500 milliunits/ml). Five of the six expressers produced FVIII and survived an otherwise lethal tail-clipping, demonstrating phenotypic correction of the bleeding disorder. FVIII expression was sustained for >14 months. Gene transfer occurred into liver, spleen, and lungs with predominant FVIII mRNA expression in the liver. Six of the seven animals with transient or no detectable human FVIII developed FVIII inhibitors (7-350 Bethesda units/ml). These findings indicate that a genetic disease can be corrected by in vivo gene therapy using retroviral vectors.


Learning our way into the future public health: a proposition.
This article attempts to bridge the gap between the values and skills that currently inform public health and those that we need to confront the future. We draw on a set of radical arguments. Firstly, the ability of modern people to understand, predict and control the natural world has brought many benefits but evidence is accumulating that the methods and mindsets of modernity are subject to diminishing returns and adverse effects. This is manifest in the rise of new epidemics: obesity, addiction-related harm, loss of well-being, rising rates of depression and anxiety and widening inequalities. Secondly, there is little evidence that people are embracing new forms of thinking or practice, despite other threats which have the potential for massive effects on many lives, such as climate change and peak oil. Thirdly, if the problems we face may indicate that 'modernity' is in decline because unsustainable, then profound change is necessary if we are to avoid collapse. This analysis suggests that public health needs a new approach. We set out propositions and models that could help us learn our way into the future.


Precision Medicine Approach for Cardiometabolic Risk Factors in Therapeutic Apheresis.
Lipoprotein apheresis (LA) is currently the most powerful intervention possible to reach a maximal reduction of lipids in patients with familial hypercholesterolemia and lipoprotein(a) hyperlipidemia. Although LA is an invasive method, it has few side effects and the best results in preventing further major cardiovascular events. It has been suggested that the highly significant reduction of cardiovascular complications in patients with severe lipid disorders achieved by LA is mediated not only by the potent reduction of lipid levels but also by the removal of other proinflammatory and proatherogenic factors. Here we performed a comprehensive proteomic analysis of patients on LA treatment using intra-individually a set of differently sized apheresis filters with the INUSpheresis system. This study revealed that proteomic analysis correlates well with routine clinical chemistry in these patients. The method is eminently suited to discover new biomarkers and risk factors for cardiovascular disease in these patients. Different filters achieve reduction and removal of proatherogenic proteins in different quantities. This includes not only apolipoproteins, C-reactive protein, fibrinogen, and plasminogen but also proteins like complement factor B (CFAB), protein AMBP, afamin, and the low affinity immunoglobulin gamma Fc region receptor III-A (FcγRIIIa) among others that have been described as atherosclerosis and metabolic vascular diseases promoting factors. We therefore conclude that future trials should be designed to develop an individualized therapy approach for patients on LA based on their metabolic and vascular risk profile. Furthermore, the power of such cascade filter treatment protocols may improve the prevention of cardiometabolic disease and its complications.


Enhanced metallosorption of Escherichia coli cells due to surface display of beta- and alpha-domains of mammalian metallothionein as a fusion to LamB protein.
The lamB gene was inserted at with DNA fragments encoding N-terminal beta- and C-terminal alpha-domains of human metallothionein 1A (HMT1A). The hybrid LamB proteins were expressed as full-length products. Virtually whole pool of hybrid LamB proteins was found localized in the outer membrane of E. coli to and cells expressing LamB variants retained sensitivity to lambda phage, indicating their correct folding. Expression of hybrid LamB proteins increased natural ability of E. coli accumulate bivalent heavy metals ions with the highest efficiency observed for cadmium. The order of amount of cadmium accumulated is alpha-domain of HMT1A > HMT1A >> beta-domain of HMT1A. This correlates with affinity for cadmium and stability of metallothionein and its individual domains. This confirms suitability of LamB vehicle for surface display of various bioactive molecules and suggests possibility of engineering of cell surface for bioremediation of heavy metals.


2019 Chinese expert consensus statement on left atrial appendage closure in patients with atrial fibrillation.
The left atrial appendage closure (LAAC), the efficacy and safety of which has been proved by a number of randomized controlled trials and registries, is recommended by several guidelines to prevent stroke in high-risk patients with non-valvular atrial fibrillation. However, current guidelines only discuss the indications and contraindications of LAAC, as an emerging technology, there still lacks comprehensive recommendations involved with LAAC, including devices, image assessment modality, identification and treatment of complications, perioperative medication, and postoperative management. Therefore, the Chinese Society of Cardiology (CSC) of Chinese Medical Association (CMA) and the Editorial Board of Chinese Journal of Cardiology jointly issued the expert consensus statement on LAAC in the prevention of stroke in patients with atrial fibrillation after comprehensive discussion by experts with different backgrounds. This consensus provided three levels of recommendations to guide and standardize the clinical application of LAAC based on existing evidence and clinical practice experience, including appropriate (more potential benefits or fewer harms), uncertain (somehow reasonable but need more evidence), and inappropriate (unlikely to benefit, or have more complications).


A Programmable Oxygenation Device Facilitates Oxygen Generation and Replenishment to Promote Wound Healing.
Inadequate oxygenation is one of the chief culprits for delayed wound healing. However, current oxygen therapies, such as hyperbaric oxygen therapy and topical oxygen therapy, face hurdles in providing sustained and long-term oxygenation to reverse wound hypoxia. Furthermore, their efficacy in rejuvenating wound injury is restricted by limited penetration of oxygen in the wound bed. Herein, this study proposes a programmable and portable oxygenation device (named GUFO oxydevice) by ingeniously integrating i) a controllable oxygen generation and unidirectional transmission system (COGT-UTS), and ii) a supramolecular assembled perfluorinated hyperbranched polymer/gelatin (GUF) hydrogel in which the perfluorinated hyperbranched polymer (FHBP) acts as an oxygen reservoir to ensure sustained and convenient oxygen replenishment and thus directly regulate the hypoxic wound microenvironment. Accelerating the wound healing process by GUFO oxydevice is achieved in both a diabetic rat and an acute porcine wound model without any secondary tissue damages. The present study demonstrates that the GUFO oxydevice holds promise as a practically feasible candidate for wound treatment.


Visualization of BRI1 and BAK1(SERK3) membrane receptor heterooligomers during brassinosteroid signaling.
The leucine-rich repeat receptor-like kinase BRASSINOSTEROID-INSENSITIVE1 (BRI1) is the main ligand-perceiving receptor for brassinosteroids (BRs) in Arabidopsis (Arabidopsis thaliana). Binding of BRs to the ectodomain of plasma membrane (PM)-located BRI1 receptors initiates an intracellular signal transduction cascade that influences various aspects of plant growth and development. Even though the major components of BR signaling have been revealed and the PM was identified as the main site of BRI1 signaling activity, the very first steps of signal transmission are still elusive. Recently, it was shown that the initiation of BR signal transduction requires the interaction of BRI1 with its SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE (SERK) coreceptors. In addition, the resolved structure of the BRI1 ectodomain suggested that BRI1-ASSOCIATED KINASE1 [BAK1](SERK3) may constitute a component of the ligand-perceiving receptor complex. Therefore, we investigated the spatial correlation between BRI1 and BAK1(SERK3) in the natural habitat of both leucine-rich repeat receptor-like kinases using comparative colocalization analysis and fluorescence lifetime imaging microscopy. We show that activation of BR signaling by exogenous ligand application resulted in both elevated colocalization between BRI1 and BAK1(SERK3) and an about 50% increase of receptor heterooligomerization in the PM of live Arabidopsis root epidermal cells. However, large populations of BRI1 and BAK1(SERK3) colocalized independently of BRs. Moreover, we could visualize that approximately 7% of the BRI1 PM pool constitutively heterooligomerizes with BAK1(SERK3) in live root cells. We propose that only small populations of PM-located BRI1 and BAK1(SERK3) receptors participate in active BR signaling and that the initiation of downstream signal transduction involves preassembled BRI1-BAK1(SERK3) heterooligomers.


Phenylalanine containing hydrophobic nanospheres for antibody purification.
In this study, novel hydrophobic nanospheres with an average size of 158 nm utilizing N-methacryloyl-(L)-phenylalanine methyl ester (MAPA) as a hydrophobic monomer were produced by surfactant free emulsion polymerization of 2-hydroxyethyl methacrylate (HEMA) and MAPA conducted in an aqueous dispersion medium. MAPA was synthesized using methacryloyl chloride and L-phenylalanine methyl ester. Specific surface area of the nonporous nanospheres was found to be 1874 m2/g. Poly(HEMA-MAPA) nanospheres were characterized by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). Average particle size, size distribution, and surface charge measurements were also performed. Elemental analysis of MAPA for nitrogen was estimated as 0.42 mmol/g polymer. Then, poly(HEMA-MAPA) nanospheres were used in the adsorption of immunoglobulin G (IgG) in batch system. Higher adsorption values (780 mg/g) were obtained when the poly (HEMA-MAPA) nanospheres were used from both aqueous solutions and human plasma. The adsorption phenomena appeared to follow a typical Langmuir isotherm. It was observed that IgG could be repeatedly adsorbed and desorbed without significant loss in adsorption amount. These findings show considerable promise for this material as a hydrophobic support in industrial processes.


Tracheal stenosis mimicking severe acute asthma.
Post-intubation tracheal stenosis is a rare but serious clinical entity that rarely develops when intubation is less than a week. These patients may remain asymptomatic for a variable period and are often misdiagnosed as asthmatic. The authors report a case of a middle-aged lady who was initially misdiagnosed as having acute asthma after brief tracheal intubation.


Mutation in the LPS outer core biosynthesis gene, galU, affects LPS interaction with the RTX toxins ApxI and ApxII and cytolytic activity of Actinobacillus pleuropneumoniae serotype 1.
Lipopolysaccharides (LPS) and Apx toxins are major virulence factors of Actinobacillus pleuropneumoniae, a pathogen of the respiratory tract of pigs. Here, we evaluated the effect of LPS core truncation in haemolytic and cytotoxic activities of this microorganism. We previously generated a highly attenuated galU mutant of A. pleuropneumoniae serotype 1 that has an LPS molecule lacking the GalNAc-Gal II-Gal I outer core residues. Our results demonstrate that this mutant exhibits wild-type haemolytic activity but is significantly less cytotoxic to porcine alveolar macrophages. However, no differences were found in gene expression and secretion of the haemolytic and cytotoxic toxins ApxI and ApxII, both secreted by A. pleuropneumoniae serotype 1. This suggests that the outer core truncation mediated by the galU mutation affects the toxins in their cytotoxic activities. Using both ELISA and surface plasmon resonance binding assays, we demonstrate a novel interaction between LPS and the ApxI and ApxII toxins via the core oligosaccharide. Our results indicate that the GalNAc-Gal II-Gal I trisaccharide of the outer core is fundamental to mediating LPS/Apx interactions. The present study suggests that a lack of binding between LPS and ApxI/II affects the cytotoxicity and virulence of A. pleuropneumoniae.


Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers.
Technologies that enable targeted manipulation of epigenetic marks could be used to precisely control cell phenotype or interrogate the relationship between the epigenome and transcriptional control. Here we describe a programmable, CRISPR-Cas9-based acetyltransferase consisting of the nuclease-null dCas9 protein fused to the catalytic core of the human acetyltransferase p300. The fusion protein catalyzes acetylation of histone H3 lysine 27 at its target sites, leading to robust transcriptional activation of target genes from promoters and both proximal and distal enhancers. Gene activation by the targeted acetyltransferase was highly specific across the genome. In contrast to previous dCas9-based activators, the acetyltransferase activates genes from enhancer regions and with an individual guide RNA. We also show that the core p300 domain can be fused to other programmable DNA-binding proteins. These results support targeted acetylation as a causal mechanism of transactivation and provide a robust tool for manipulating gene regulation.


Effect of transversus abdominis plane block with or without buprenorphine after inguinal hernia surgery on postoperative pain.
Transversus abdominis plane (TAP) has been mentioned as having bene-ficial effects on chronic pain after hernioplasty. This study assessed the effects of TAP block on acute and persistent postoperative pain after inguinal hernia surgery, with or without buprenorphine. 64 patients were allocated to group R ( n = 32) and received 20 mL of 0.25% ropivacaine for TAP block; group RB ( n = 32) received 20 mL of 0.25% ropivacaine containing 300 µg of buprenorphine for TAP block. The primary outcome was the analgesic and antihyperalgesic effect of buprenorphine. The duration of analgesia, analgesic consumption, postoperative pain scores at rest and sitting up to 48 hours, and the effect on wound hyperalgesia were evaluated. Secondary outcomes included the incidence of side effects and complications. The median (IQR) duration of analgesia in group R was 386.5 (37.25) minutes vs. 868 (41.3) minutes in the RB group. Median pain scores on sitting were found to be significantly better in group RB than in group R at 6, 12, and 24 hours ( P < 0.001). The wound hyperalgesia index showed a significant difference between groups ( P < 0.001). The incidence of persistent postoperative pain was 6.25% in the R group, as compared to 0% in the RB group. Otherwise, the patients did not have any further complications associated with the block. The results demonstrated that TAP block with buprenorphine reduced acute postoperative pain severity, but we did not find a difference between groups in persistent pain.


Type-1 interferons prolong the lifespan of neutrophils by interfering with members of the apoptotic cascade.
Polymorphonuclear Neutrophils (PMNs) are metabolically highly active phagocytes, present in abundant numbers in the circulation. These active cells take the onus of clearing invading pathogens by crowding at inflammatory sites in huge numbers. Though PMNs are extremely short living and die upon spontaneous apoptosis, extended lifespan has been observed among those cells arrive at the inflammation sites or tackle intracellular infections or face any microbial challenges. The delay/inhibition of spontaneous apoptosis of these short-living cells at the inflammatory core rather helps in combating pathogens. Like many candidates, type-1 interferons (type-1 IFNs) is a group of cytokines predominant at the inflammation site. Although there are some isolated reports, a systematic study is still lacking which addresses the impact of the predominant type of interferon on the spontaneous apoptosis of neutrophils. Here in, we have observed that exposure of these IFNs (IFN-β, IFN-α & IFN-ω etc) on human neutrophils prevents the degradation of the Bfl1, an important anti-apoptotic partner in the apoptotic cascade. Treatment showed a significant reduction in the release of cytochrome-C in the cytosol, a critical regulator in the intrinsic apoptotic pathway. We also noticed a reduction in the conversion of procaspase -3 to active caspase-3, a crucial executioner caspase towards initiation of apoptosis. Taken together our results show that exposure to interferon interferes with apoptotic pathways of neutrophils and thereby delay its spontaneous apoptosis. These findings would help us further deciphering specific roles if these inflammatory agents are causing any immune-metabolomic changes on PMNs at the inflammatory and infection core.


Ultrasound-based stereotactic guidance in prostate cancer--quantification of organ motion and set-up errors in external beam radiation therapy.
A mobile transabdominal ultrasound-based targeting system (BAT(R)) has been developed which can stereotactically localize the position of the prostate each treatment day and directly integrate this information into the treatment planning system. Daily target verification facilitates a marked reduction in planning treatment margins by correcting potential organ-motion and set-up errors. Previous studies have been performed to establish the precision of ultrasound localization. This report quantifies the magnitude of the patient isocenter shift parameters encountered during clinical implementation of this system. After five weeks of conformal external beam radiation therapy, 54 patients underwent a second CT simulation. Prostate-only fields based on this scan were created with no PTV margin beyond the CTV. For each of the final conedown treatments (2-4 fractions), patients underwent ultrasound-based stereotactic prostate localization at the treatment machine. The portable system, which electronically imports the CT simulation target-contour and isocenter information, is situated adjacent to the treatment couch. Transverse and sagittal suprapubic ultrasound images are captured, and the system electronically couples this data to the baseline isocenter. The CT contours are maneuvered in three dimensions by a touch-screen menu to visually overlay the ultrasound images. The system then displays the three-dimensional (3D) couch shifts required to produce field alignment. One hundred and eighty-nine daily ultrasound prostate position shifts were recorded for 54 patients. The isocenter field misalignment between the baseline CT and ultrasound ranged from -26.8 to 33.8 mm in the anterior/posterior (A/P) dimension, -10.2 to 30.9 mm in the lateral dimension, and -24.6 to 9.0 mm in the superior/inferior (S/I) dimension. The corresponding directed average disagreements were -3.0 mm (SD 8.3 mm) A/P, 1.86 mm (SD 5.7 mm) lateral, and -2.6 mm (SD 6.5 mm) S/I. The magnitudes of undirected misalignments were frequently larger than 5 mm (51% of A/P, 31% of lateral, and 35% of superior measurements) and oftentimes larger than 10 mm (21% of A/P, 7% of lateral, and 12% of superior measurements). Organ motion and set-up uncertainties limit optimization of 3D treatment planning by expanding the width of PTV margins required to ensure target coverage. Transabdominal ultrasound-based stereotactic guidance is a safe and direct method for correcting patient positioning. Our experience with the BAT system in a large cohort of prostate cancer patients revealed that substantial daily isocenter corrections were encountered in a large percentage of cases. This data would suggest that daily clinical isocenter misalignments are greater than would be expected from published data on organ motion and set-up variations encountered in the study setting.


Comparison of Fluid-Fluid Interfacial Areas Measured with X-ray Microtomography and Interfacial Partitioning Tracer Tests for the same Samples.
Two different methods are currently used for measuring interfacial areas between immiscible fluids within 3-D porous media, high-resolution microtomographic imaging and interfacial partitioning tracer tests (IPTT). Both methods were used in this study to measure non-wetting/wetting interfacial areas for a natural sand. The microtomographic imaging was conducted on the same packed columns that were used for the IPTTs. This is in contrast to prior studies comparing the two methods, for which in all cases different samples were used for the two methods. In addition, the columns were imaged before and after the IPTTs to evaluate the potential impacts of the tracer solution on fluid configuration and attendant interfacial area. The interfacial areas measured using IPTT are ~5 times larger than the microtomographic-measured values, which is consistent with previous work. Analysis of the image data revealed no significant impact of the tracer solution on NAPL configuration or interfacial area. Other potential sources of error were evaluated, and all were demonstrated to be insignificant. The disparity in measured interfacial areas between the two methods is attributed to the limitation of the microtomography method to characterize interfacial area associated with microscopic surface roughness due to resolution constraints.


Schizophrenia and substance abuse comorbidity: nicotine addiction and the neonatal quinpirole model.
This review focuses on nicotine comorbidity in schizophrenia, and the insight into this problem provided by rodent models of schizophrenia. A particular focus is on age differences in the response to nicotine, and how this relates to the development of the disease and difficulties in treatment. Schizophrenia is a particularly difficult disease to model in rodents due to the fact that it has a plethora of symptoms ranging from paranoia and delusions of grandeur to anhedonia and negative affect. The basis of these symptoms is believed to be due to neurochemical abnormalities and neuropathology in the brain, which most models have attempted to emulate. A brief review of findings regarding nicotine use and abuse in schizophrenics is presented, with findings using rodent models that have been able to provide insight into the mechanisms of addiction. A common clinical approach to the treatment of nicotine addiction in the schizophrenic population has been that these drugs are used for self-medication purposes, and it is clear that self-medication may actually be directed at several symptoms, including cognitive impairment and anhedonia. Finally, our laboratory has reported across a series of studies that neonatal treatment with the dopamine D(2)/D(3) receptor agonist quinpirole results in long-term increases in dopamine-like receptor sensitivity, consistent with data reporting increases in dopamine D(2) receptor function in schizophrenia. Across these studies, we have reported several behavioral, neurochemical, and genetic consistencies with the disease, and present a hypothesis for what we believe to be the basis of psychostimulant addiction in schizophrenia.


[Erectile dysfunction after radial prostatectomy. Response to treatment with sildenafil citrate].
Erectile dysfunction is one of the sequelae after radical prostatectomy most concerning for the patient, its appearance being influenced by factors such as surgical technique, or patient's age. Thus, around 40% of the patients younger than 70 years and over 90% of the patients older than 70 present with erectile dysfunction after prostatectomy. We studied the response to sildenafil citrate in patients who underwent non nerve sparing radical prostatectomy. Medical records of 100 patients who underwent radical prostatectomy at our department between 1985 and 1998 were retrospectively reviewed and telephone interviews were done to investigate the degree of erectile dysfunction following surgery. All patients were offered to start a two-month therapeutic trial with sildenafil citrate, in dose escalation up to a maximum of 100 mg. Therapeutic response was evaluated by personal interviews. 5 out of 100 patients interviewed showed similar potency than preoperatively. Among 95 patients with erectile dysfunction 63 (66.3%) did not wish treatment for erectile dysfunction. 22 of the remainder 32 patients initiated treatment with sildenafil citrate, whereas 10 preferred trying intracavernous PGE1 injections only. In the intracavernous injections group 5 patients abandoned because of pain with injections and refused any further treatment with oral drugs; 5 patients are satisfied with intracavernous therapy and they do not want to try sildenafil. In the sildenafil group form 22 patients starting treatment 14 (63.6%) are satisfied, with a subjective improvement. Only 5 patients (22%) show an erectile response enough to have sexual intercourse with penetration. 8 (36.5%) abandon treatment because of lack of efficiency. 5% of the patients who undergo non nerve sparing radical prostatectomy maintain sexual potency without treatment postoperatively. Most patients with postoperative erectile dysfunction (63/95 = 66.3%) do not want any treatment. From 22 patients initiating treatment with sildenafil 5 can have sexual intercourse with penetration (5/22 = 22%), all of them with 100 mg doses of sildenafil.


Transient population dynamics in flashlamp pumped sensitized erbium glass lasers.
Erbium population dynamics in ytterbium sensitized phosphate glass is studied by measuring transient changes in laser probe transmission during flashlamp pumping. The influence of energy loss channels such as excitation cumulation and nonlinear fluorescence quenching on the flashlamp pumping efficiency is observed to be relatively small. The decrease of the pumping efficiency at high input energies can be attributed mostly to the blue shift in the flashlamp output radiation spectrum.


Genome-wide association study in RPGRIP1(-/-) dogs identifies a modifier locus that determines the onset of retinal degeneration.
Cone-rod dystrophy (CRD) is a form of inherited retinal degeneration (RD) causing blindness in man as well as in several breeds of dog. Previously, a 44 bp insertion in RPGRIP1 (retinitis pigmentosa GTPase regulator interacting protein-1) was associated with a recessive early-onset CRD (cone-rod dystrophy 1, cord1) in a Miniature longhaired dachshund (MLHD) research colony. Yet in the MLHD pet population, extensive range of the onset age has been observed among RD cases, with some RPGRIP1(-/-) dogs lacking obvious clinical signs. Phenotypic variation has been known in human homologous diseases, including retinitis pigmentosa and Leber congenital amaurosis, indicating possible involvement of modifiers. To explore additional genetic loci associated with the phenotypic variation observed in MLHDs, a genome-wide association study was carried out using Canine SNP20 arrays in 83 RPGRIP1(-/-) MLHDs with variable ages of onset or no clinical abnormality. Using these samples, comparison of 31 early-onset RD cases against 49 controls (15 late-onset RD and 34 normal dogs combined) identified a strong association (P = 5.05 × 10(-13)) at a single locus on canine chromosome 15. At this locus, the majority of early-onset RD cases but few of the controls were homozygous for a 1.49 Mb interval containing ~11 genes. We conclude that homozygosity at both RPGRIP1 and the newly mapped second locus is necessary to develop early-onset RD, whereas RPGRIP1(-/-) alone leads to late-onset RD or no apparent clinical phenotype. This study establishes a unique model of canine RD requiring homozygous mutations at two distinct genetic loci for the manifestation of early-onset RD.


The acute effects of acrivastine (BW825C), a new antihistamine, compared with triprolidine on measures of central nervous system performance and subjective effects.
The new H1-antagonist acrivastine (BW825C) in doses of 4, 8, and 16 mg was compared with triprolidine HCl (2.5 and 5 mg) in a double-blind crossover study in 12 subjects. Adaptive tracking performance 1.5 hours after dosing was impaired by triprolidine, 2.5 and 5 mg; the impairment was still detectable 3.5 hours after 5 mg. Acrivastine did not impair adaptive tracking after any of the doses. Triprolidine increased reaction times after 1.5 hours (2.5 and 5 mg) and 3 hours (5 mg), but acrivastine did not have any effect on reaction time at any dose. Both doses of triprolidine caused subjective central nervous system effects after 1.5 hours, and triprolidine, 5 mg, still had some detectable effects on subjective rating scales after 3 hours. No subjective effects were noted after acrivastine. We conclude that acrivastine at doses causing more peripheral H1-antagonism than triprolidine has considerably reduced central nervous system activity.


[Invasive pulmonary aspergillosis in patients with acute respiratory syndrome by COVID-19].
Patients with COVID-19 who are admitted to intensive care unit (ICU) are at high risk of developing secondary infections, including invasive fungal infections such as invasive pulmonary aspergillosis (IPA). The main purpose was to analyse the putative COVID-19 Associated Pulmonary Aspergillosis (CAPA) patients in our setting. In these patients, we performed mycological culture in bronchoalveolar lavage (BAL) for isolation of Aspergillus sp. We followed the AspICU algorithm to diagnose putative IPA. Moreover, we considered relevant the positivity of galactomannan in BAL. We diagnosed putative IPA in 3 patients. The common features of these 3 patients were: more than 21 days of stay in ICU, severe acute respiratory distress syndrome (ARDS) and treatment with steroids (1 mg/kg per day). Therefore, CAPA has to be systematically considered although a new algorithm to diagnose it is needed to treat patients in early stages in order to avoid catastrophic outcomes.


Genome Wide Analysis Reveals the Extrinsic Cellulolytic and Biohydrogen Generating Abilities of Neocallimastigomycota Fungi.
Ruminating animals, especially cattle lack the carbohydrate active enzyme encoding genes which are required for the degradation of the glycosidic linkages of plant cell wall carbohydrates (such as cellulose, hemicellulose, lignin and pectin). Thus, ruminating animals are completely dependent on the microorganisms (anaerobic bacteria and fungi, methanogenic archaea and protozoa) residing in their rumen (hindgut). In this study, we have retrieved and analyzed the complete genome wide annotations of the Neocallimastigomycota division fungi such as Anaeromyces robustus, Neocallismatix californiae, Orpinomyces sp, Piromyces finnis, Piromyces sp E2. We have retrieved the InterPro, CAZy, KOG, KEGG, SM Clusters and MEROPS genome level data of these anaerobic fungi from JGI-MycoCosm database. Results obtained in our study reveals that, the genomes of anaerobic fungi completely lack genes encoding for lignin degrading auxiliary activity enzymes. Contrastingly, these fungi outnumbered other fungi by having highest number of CAZyme encoding genes. The genes encoding for dockerins and carbohydrate binding modules exaggerated other CAZymes which are involved in the structure and functioning of cellulosomes. Presence of cellulosomes and higher number of carbohydrate transport and metabolism genes also endorses the plant cell wall carbohydrate degrading abilities of these fungi. We also reported the tentative total cellulolytic, hemicellulolytic and pectinolytic abilities. And we have explicitly reported the genes, enzymes and the mechanisms involved in structure and functioning of the cellulosomes and hydrogenosomes. Our present work reveals the genomic machinery underlying the extrinsic plant cell wall degrading abilities of the anaerobic fungi. Results obtained in our study can be significantly applied in improving the gut health of cattle and especially in the fields of biofuel, biorefining and bioremediation-based industries.


Effects of Systemic Lidocaine on Postoperative Recovery Quality and Immune Function in Patients Undergoing Laparoscopic Radical Gastrectomy.
This study aimed to explore the effects of lidocaine on postoperative quality of recovery (QoR) and immune function in patients undergoing laparoscopic radical gastrectomy. In total, 135 patients were enrolled and were equally randomized to receive low-dose lidocaine (Group LL: 1.5 mg/kg bolus followed by an infusion at 1.0 mg/kg/hour) or high-dose lidocaine (Group HL: 1.5 mg/kg bolus followed by an infusion at 2.0 mg/kg/hour) or Controls (Group C: received a volume-matched normal saline at the same rate). The primary outcome was a QoR-40 score on postoperative day (POD) 1. Secondary outcomes were a QoR-40 score on POD 3, levels of inflammatory factors (IL-6, IL-10, TNF-α) and CD4+T cells, CD8+T cells proportions, and CD4+/CD8+ cell ratios and postoperative recovery of bowel function. There were no statistically significant differences in patient characteristics at baseline. The total QoR-40 scores on POD 1 in Group HL (171.4±3.89) were higher than those in Group LL (166.20±4.05) and in Group C (163.40±4.38) (adjusted P<0.001). Differences in the dimension scores of QoR-40 for pain, physical comfort, and emotional state were significant across the three groups. Lidocaine administration significantly reduced the release of IL-6, IL-10, TNF-α, and attenuated immune changes induced by trauma. Kaplan-Meier curves showed that the median time to the first exhaust and defecation were shorter in the Group HL than in Groups LL and C (1.55 days vs 2.4 days vs 2.6 days, log rank P<0.0001; and 2.86 days vs 3.22 days vs 3.46 days, log rank P=0.002, respectively). Additionally, patients in lidocaine groups required less remifentanil consumption and experienced lower pain intensity, compared with the control group. Systemic lidocaine improved postoperative recovery, alleviated inflammation and immunosuppression, and accelerated the return of bowel function, and is thus, worthy of clinical application. ChiCTR2000028934.


Responses of birds and mammals to long-established wind farms in India.
Wind turbines have been recognised as an alternative and clean-energy source with a low environmental impact. The selection of sites for wind-farm often creates serious conservation concerns on biodiversity. Wind turbines have become a serious threat to migratory birds as they collide with the turbine blades in some regions across the globe, while the impact on terrestrial mammals is relatively less explored. In this context, we assessed the responses of birds and mammals to the wind turbines in central Karnataka, India from January 2016 to May 2018 using carcass searches to quantify animal collisions (i.e., birds and bats), fixed radius point count for bird population parameters, and an occupancy framework for assessing the factor that determines the spatial occurrence of terrestrial mammals. The mean annual animal fatality rate per wind turbine was 0.26/year. Species richness, abundance, and unique species of birds were relatively higher in control sites over wind turbine sites. Species and functional compositions of birds in control sites were different from wind turbine sites, explaining the varied patterns of bird assemblages of different feeding guilds. Blackbuck, Chinkara, Golden Jackal, and Jungle Cat were less likely to occupy sites with a high number of wind turbines. The study indicates that certain bird and mammal species avoided wind turbine-dominated sites, affecting their distribution pattern. This is of concern to the management of the forested areas with wind turbines. We raised conservation issues and mitigating measures to overcome the negative effects of wind turbines on animals.


The clinical profile of amblyopia in children younger than 3 years of age.
Amblyopia in children ≥3 years has been well described, but less is known about amblyopia in children <3 years of age. Here we describe the clinical characteristics of a large cohort of children <3 years of age with amblyopia and compare them with a previously described Pediatric Eye Disease Investigator Group amblyopic cohort aged 3 to 6 years. A total of 250 consecutive children with amblyopia <3 years were referred by 16 pediatric ophthalmologists. The mean age at the initial diagnosis of amblyopia was 1.2 ± 0.7 years. The cause of amblyopia was strabismus in 82%, anisometropia in 5%, and combined mechanism in 13%. Compared with the 3- to 6-year-old cohort, the proportion of amblyopia attributable to strabismus was significantly greater (p < 0.001), whereas both anisometropia and combined mechanism amblyopia were significantly less common (p < 0.001). Overall, 61% of amblyopia was diagnosed at the same visit during which strabismus and/or anisometropia was initially diagnosed; an additional 21% of amblyopia was diagnosed at the first follow-up visit 1 to 3 months later. Compared with the 3- to 6-year-old cohort, amblyopic eye refractive error was significantly lower. Strabismic amblyopia was diagnosed much more commonly than anisometropic and combined-mechanism amblyopia in children <3 years. Anisometropic amblyopia may be difficult to detect in children <3 years, and/or strabismic amblyopia may be overdiagnosed by fixation preference. Alternatively, anisometropia may develop more commonly after 3 years of age or may require greater duration to cause amblyopia.


The prevalence and incidence of sick building syndrome in Chinese pupils in relation to the school environment: a two-year follow-up study.
There are few incidence studies on sick building syndrome (SBS). We studied two-year change of SBS in Chinese pupils in relation to parental asthma/allergy (heredity), own atopy, classroom temperature, relative humidity (RH), absolute humidity (AH), crowdedness, CO₂, NO₂, and SO₂. A total of 1993 participated at baseline, and 1143 stayed in the same classrooms after two years. The prevalence of mucosal and general symptoms was 33% and 28% at baseline and increased during follow-up (P < 0.001). Twenty-seven percent reported at least one symptom improved when away from school. Heredity and own atopy were predictors of SBS at baseline and incidence of SBS. At baseline, SO₂ was associated with general symptoms (OR=1.10 per 100 μg/m³), mucosal symptoms (OR=1.12 per 100 μg/m³), and skin symptoms (OR=1.16 per 100 μg/m³). NO₂ was associated with mucosal symptoms (OR=1.13 per 10 μg/m³), and symptoms improved when away from school (OR=1.13 per 10 μg/m³). Temperature, RH, AH, and CO₂ were negatively associated with prevalence of SBS. Incidence or remission of SBS was not related to any exposure, except a negative association between SO₂ and new skin symptoms. In conclusion, heredity and atopy are related to incidence and prevalence of SBS, but the role of the measured exposures for SBS is more unclear. We found high levels of CO₂ indicating inadequate ventilation and high levels of SO₂ and NO₂, both indoors and outdoors. All schools had natural ventilation, only. Relying on window opening as a tool for ventilation in China is difficult because increased ventilation will decrease the level of CO₂ but increase the level of NO₂ and SO₂ indoors. Prevalence studies of sick building syndrome (SBS) might not be conclusive for causal relationships, and more longitudinal studies on SBS are needed both in China and other parts of the world. The concept of mechanical ventilation and air filtration should be introduced in the schools, and when planning new schools, locations close to heavily trafficked roads should be avoided.


Individuals with hemiparetic stroke abnormally perceive their elbow torques when abducting their paretic shoulder.
Individuals with hemiparetic stroke exhibit an abnormal coupling between shoulder abduction and elbow flexion, or flexion synergy, due to an increased reliance on cortico-bulbospinal pathways. While this motor impairment is well documented, its impact on how movements are perceived remains unexplored. This study investigates whether individuals with hemiparetic stroke accurately perceive torques at their paretic elbow while abducting at their shoulder. Ten individuals with hemiparetic stroke participated. We recorded the extent of their abnormal joint coupling as the torque at their elbow, with respect to the maximum voluntary torque in elbow flexion, when abducting at their shoulder. Next, we estimated the perception of their elbow torque by reporting their errors on our torque-matching task. When abducting at the shoulder, the participants with stroke generated a greater non-volitional torque at their paretic elbow (13.2 ± 8.7%) than their non-paretic elbow (1.2 ± 11.2%) (p = 0.003). Regarding the perception of our torque-matching task, participants overestimated their torques to a lesser extent at their paretic elbow (1.8 ± 6.6%) than at their non-paretic elbow (6.2 ± 5.4%) (p = 0.004). Torque perception at the paretic elbow differed from the non-paretic elbow when abducting at the shoulder. This work advances our understanding of the i) somatosensory deficits occurring post hemiparetic stroke and ii) neural basis of torque perception.


Crystallization, structural determination and analysis of a novel parasite vaccine candidate: Fasciola hepatica glutathione S-transferase.
Glutathione S-transferases (GSTs) represent the major class of detoxifying enzymes from parasitic helminths. As a result, they are candidates for chemotherapeutic and vaccine design. Indeed, GSTs from Fasciola hepatica have been found to be effective for vaccinating sheep and cattle against fasciolosis. This helminth contains at least seven GST isoforms, of which four have been cloned. The cloned isoforms (Fh51, Fh47, Fh7 and Fh1) all belong to the mu class of GSTs, share greater than 71% sequence identity, yet display distinct substrate specificities. Crystals of Fh47 were obtained using the hanging drop vapour diffusion technique. The crystals belong to space group I4122, with one monomer in the asymmetric unit, which corresponds to a very high solvent content of approximately 75%. The physiological dimer is generated via a crystallographic 2-fold rotation. The three-dimensional structure of Fh47 was solved by molecular replacement using the Schistosoma japonicum glutathione S-transferase (Sj26) crystal structure as a search model. The structure adopts the canonical GST fold comprising two domains: an N-terminal glutathione-binding domain, consisting of a four-stranded beta-sheet and three helices whilst the C-terminal domain is entirely alpha-helical. The presence of Phe19 in Fh47 results in a 6 degrees interdomain rotation in comparison to Sj26, where the equivalent residue is a leucine. Homology models of Fh51, Fh7 and Fh1, based on the Fh47 crystal structure, reveal critical differences in the residues lining the xenobiotic binding site, particularly at residue positions 9, 106 and 204. In addition, differences amongst the isoforms in the non-substrate binding site were noted, which may explain the observed differential binding of large ligands. The major immunogenic epitopes of Fh47 were surprisingly found not to reside on the most solvent-exposed regions of the molecule.


The effect of whitening agents on caries susceptibility of human enamel.
This in vitro study evaluated whether the treatment of human enamel with whitening agents containing different concentrations of carbamide or hydrogen peroxide changes the susceptibility of enamel to caries. Twenty-four sound human incisors were selected for this study. For each tooth, the crown was sectioned into two halves in the cervical-incisal direction. One half of the sectioned tooth was treated and the other half was used as a control specimen. Each half was randomly divided into three treatment groups (eight two-halves/group). The whitening agents were 10% carbamide peroxide, 20% carbamide peroxide with fluoride and 35% hydrogen peroxide. Following pretreatment, the specimens were demineralized for four days in an in vitro microbial caries model and then analyzed using a confocal laser scanning microscope (CLSM). Results showed that there were no significant differences between the treated and controlled specimens for teeth treated with 10% carbamide peroxide or 35% hydrogen peroxide. However, specimens treated with 20% carbamide peroxide with FP (0.11% fluoride and potassium nitrate) were less susceptible to caries than their controls at p < or = 0.05. In conclusion, application of bleaching agents does not increase the caries susceptibility of human enamel.


Clinical and Biological Characteristics of 40 Patients With Neurosyphilis and Evaluation of Treponema pallidum Nested Polymerase Chain Reaction in Cerebrospinal Fluid Samples.
Syphilis remains a significant public health problem. We conducted a prospective study to define more precisely the clinical and biological characteristics of patients with neurosyphilis (NS), and we assessed the diagnostic value of nested polymerase chain reaction (PCR) testing for Treponema pallidum in cerebrospinal fluid (CSF) samples. From 2001 to 2013, we included 40 patients (90% men; 45% infected with human immunodeficiency virus) with NS, defined as syphilis with neurological and/or ophthalmological symptoms and CSF abnormalities. Thirty patients (75%) had early, 5 (12.5%) had late, and 5 had meningovascular NS. Twenty-four patients (80%) with early NS had ophthalmological symptoms, 14 (47%) had neurological symptoms, and 8 (26%) had both. All patients with meningovascular NS had only neurological symptoms. All patients with late NS had neurological symptoms, and 2 (40%) also had ocular symptoms. Ophthalmological symptoms were present in 65% of all patients with NS, and neurological symptoms in 60%. Seventeen patients (42.5%) had CSF white blood cell counts >20/μL (mean, 57/μL), and 27 (67.5%) had high CSF protein levels (>0.5 g/L; mean value, 1 g/L). CSF PCR results were positive in 42%, and CSF VDRL results in 30%. The nested PCR assay had an overall sensitivity of 42.5%, a specificity of 97%, a positive predictive value of 77%, and a negative predictive value of 86%. Early NS is the most frequent presentation, with an overrepresentation of polymorphous ophthalmological symptoms. PCR is highly specific and of potential value when used with other biological parameters.


Wide Field Spectral Imaging with Shifted Excitation Raman Difference Spectroscopy Using the Nod and Shuffle Technique.
Wide field Raman imaging using the integral field spectroscopy approach was used as a fast, one shot imaging method for the simultaneous collection of all spectra composing a Raman image. For the suppression of autofluorescence and background signals such as room light, shifted excitation Raman difference spectroscopy (SERDS) was applied to remove background artifacts in Raman spectra. To reduce acquisition times in wide field SERDS imaging, we adapted the nod and shuffle technique from astrophysics and implemented it into a wide field SERDS imaging setup. In our adapted version, the nod corresponds to the change in excitation wavelength, whereas the shuffle corresponds to the shifting of charges up and down on a Charge-Coupled Device (CCD) chip synchronous to the change in excitation wavelength. We coupled this improved wide field SERDS imaging setup to diode lasers with 784.4/785.5 and 457.7/458.9 nm excitation and applied it to samples such as paracetamol and aspirin tablets, polystyrene and polymethyl methacrylate beads, as well as pork meat using multiple accumulations with acquisition times in the range of 50 to 200 ms. The results tackle two main challenges of SERDS imaging: gradual photobleaching changes the autofluorescence background, and multiple readouts of CCD detector prolong the acquisition time.


Morphological and functional cardiac alterations in children with congenital Zika syndrome and severe neurological deficits.
Zika virus infection during pregnancy causes fetal microcephaly and brain damage. Congenital Zika syndrome (CZS) is characterized by systemic involvement with diffuse muscle impairment, a high frequency of arthrogryposis, and microphthalmia. Cardiac impairment in CZS has rarely been evaluated. Our study assessed morphology and biventricular cardiac function in children with CZS and advanced neurological dysfunction. This cross-sectional study was conducted on 52 children with CZS (Zika group; ZG) and 25 healthy children (control group; CG) in Paraiba, Brazil. Clinical evaluation, electrocardiogram (EKG), and transthoracic echocardiogram (TTE) were performed on all children. Additionally, troponin I and natriuretic peptide type B (BNP) levels, the degree of cerebral palsy, and neuroimaging findings were assessed in the ZG group. The median age of the study population was 5 years in both groups, and 40.4% (ZG) and 60% (CG) were female. The most prevalent electrocardiographic alteration was sinus arrhythmia in both the ZG (n = 9, 17.3%) and CG (n = 4, 16%). The morphological parameters adjusted for Z score were as follows: left ventricular (LV) end-diastolic diameter in ZG: -2.36 [-5.10, 2.63] vs. CG: -1.07 [-3.43, 0.61], p<0.001); ascending aorta (ZG: -0.09 [-2.08, 1.60] vs. CG: 0.43 [-1.47, 2.2], p = 0.021); basal diameter of the right ventricle (RV) (ZG: -2.34 [-4.90, 0.97] vs. CG: -0.96 [-2.21, 0.40], p<0.01); and pulmonary artery dimension (ZG: -2.13 [-5.99, 0.98] vs. CG: -0.24 [-2.53, 0.59], p<0.01). The ejection fractions (%) were 65.7 and 65.6 in the ZG and CG, respectively (p = 0.968). The left atrium volume indices (mL/m2) in the ZG and CG were 13.15 [6.80, 18.00] and 18.80 [5.90, 25.30] (p<0.01), respectively, and the right atrium volume indices (mL/m2) were 10.10 [4.90, 15.30] and 15.80 [4.10, 24.80] (p<0.01). The functional findings adjusted for Z score were as follows: lateral systolic excursion of the mitral annular plane (MAPSE) (ZG: 0.36 [-2.79, 4.71] vs. CG: 1.79 [-0.93, 4.5], p = 0.001); tricuspid annular plane systolic excursion (TAPSE) (ZG: -2.43 [-5.47, 5.09] vs. CG: 0.07 [-1.98, 3.64], p<0.001); and the S' of the RV (ZG: 1.20 [3.35, 2.90] vs. CG: -0.20 [-2.15, 1.50], p = 0.0121). No differences in biventricular strain measurements were observed between the groups. Troponin I and BNP levels were normal in in the ZG. Grade V cerebral palsy and subcortical calcification were found in 88.6% and 97.22% of children in the ZG group, respectively. A reduction in cardiac dimensions and functional changes were found in CZS patients, based on the TAPSE, S' of the RV, and MAPSE, suggesting the importance of cardiac evaluation and follow-up in this group of patients.


Reproducible in vitro model for dystrophic calcification of cardiac valvular interstitial cells: insights into the mechanisms of calcific aortic valvular disease.
Calcific aortic valvular disease (CAVD) is the most prevalent valvular pathology in the United States. Development of a pharmacologic agent to slow, halt, or reverse calcification has proven to be unsuccessful as still much remains unknown about the mechanisms of disease initiation. Although in vitro models of some features of CAVD exist, their utility is limited by the inconsistency of the size and time course of the calcified cell aggregates. In this study, we introduce and verify a highly reproducible in vitro method for studying dystrophic calcification of cardiac valvular interstitial cells, considered to be a key mechanism of clinical CAVD. By utilizing micro-contact printing, we were able to consistently reproduce cell aggregation, myofibroblastic markers, programmed cell death, and calcium accumulation within aggregates of 50-400 μm in diameter on substrates with moduli from 9.6 to 76.8 kPa. This method is highly repeatable, with 70% of aggregates staining positive for Alizarin Red S after one week in culture. Dense mineralized calcium-positive nanoparticles were found within the valvular interstitial cell aggregates as shown by scanning electron microscopy (SEM) and energy dispersive spectrometry (EDS). The area of micro-contact printed aggregates staining positive for caspase 3/7 activity increased from 5.9 ± 0.9% to 12.6 ± 4.5% over one week in culture. Z-VAD-FMK reduced aggregates staining positive for Alizarin Red S by 60%. The state of cell stress is hypothesized to play a role in the disease progression; traction force microscopy indicates high substrate stresses along the aggregate periphery which can be modulated by altering the size of the aggregates and the modulus of the substrate. Micro-contact printing is advantageous over the currently used in vitro model as it allows the independent study of how cytokines, substrate modulus, and pharmacologic agents affect calcification. This controlled method for aggregate creation has the potential to be used as an in vitro assay for the screening of promising therapeutics to mitigate CAVD.


Neutrophilic dermatoses: part II.
This article addresses neutrophilic dermatoses, thus complementing the previous article (part I). The following dermatoses are introduced and discussed: subcorneal pustular dermatosis (Sneddon-Wilkinson disease), dermatitis cruris pustulosa et atrophicans, acute generalized exanthematous pustulosis, continuous Hallopeau acrodermatitis, palmoplantar pustulosis, infantile acropustulosis, Andrews' pustular bacteride and eosinophilic pustular folliculitis. A brief review of neutrophilic dermatoses in pediatric patients is also conducted.


Versatile self-complexing compounds based on covalently linked donor-acceptor cyclophanes.
A range of covalently linked donor-acceptor compounds which contain 1) a hydroquinone (HQ) unit, 2) a 1,5-dioxynaphthalene (DNP) ring system, or 3) a tetrathiafulvalene (TTF) unit as the pi-donor, and 4) cyclobis(paraquat-p-phenylene) (CBPQT(4+)) as the pi-accepting tetracationic cyclophane were prepared and shown to operate as simple molecular machines. The pi-donating arms can be included inside the cyclophane in an intramolecular fashion by virtue of stabilizing noncovalent bonding interactions. What amounts to self-complexing/decomplexing equilibria were shown to be highly temperature dependent when the pi-donating arm contains either an HQ or DNP moiety. The thermodynamic parameters associated with the equilibria have been unraveled by using variable-temperature (1)H NMR spectroscopy. The negative DeltaH degrees and DeltaS degrees values account for the fact that the "uncomplexed" conformation becomes the dominant species, since the entropy gain associated with the decomplexation process overcomes the enthalpy loss resulting from the breaking of the donor-acceptor interactions. The arm's in-and-out movements with respect to the linked cyclophanes can be arrested by installing a bulky substituent at the end of the arm. In the case of compounds carrying a DNP ring system in their side arm, two diastereoisomeric, self-complexing conformations are observed below 272 K in hexadeuterioacetone. By contrast, control over the TTF-containing arm's movement is more or less ineffective through the thermally sensitive equilibrium although it can be realized by chemical and electrochemical ways as a result of TTF's excellent redox properties. Such self-complexing compounds could find applications as thermo- and electroswitches. In addition, the thermochromism associated with the arm's movement could lead to some of the compounds finding uses as imaging and sensing materials.


Predicting domains and rates of change in borderline personality disorder.
What changes and how quickly these changes occur as a result of therapy in borderline personality disorder (BPD) is an important ongoing question. The features of BPD patients that are most predictive of rates of change in such patients remain largely unknown. Using the Cornell Personality Disorders Institute (CPDI) randomized controlled trial data, we sought to determine (a) the number and nature of broad domains underlying a large number of rate of change (slope) measures across many psychological, psychiatric, and psychosocial indexes, and (b) which baseline individual difference psychological features of the BPD patients correlated with these rate of change domains. We examined the latent structure of slope (rate of change) measures gleaned from individual growth curves for each subject, studied in multiwave perspective, on separate measures of anger, aggression, impulsivity, depression, global functioning, and social adjustment. Three broad domains of change rate could be discerned. These domains were reflected in factors that are described as (a) anger/aggression change ("aggressive dyscontrol"), (b) global functioning/social adjustment change ("social adjustment/self-acceptance"), and (c) anxiety/depression/impulsivity change ("conflict tolerance/behavioral control"). Factor scores were computed for each change domain and baseline measures of personality and psychodynamic features, selected a priori, were correlated with these factor scores. Multiple regression analyses revealed (a) baseline negative affectivity and aggression predicted the aggressive dyscontrol change domain, (b) baseline identity diffusion predicted the social adjustment/self-acceptance change domain, and (c) baseline social potency predicted the conflict tolerance/behavioral control change domain. These baseline predictors suggest potential research foci for understanding those aspects of BPD that change at comparable rates over time.


Effect of pathological fracture on limb salvage surgery with preservation of the epiphysis in children with osteosarcoma of the distal femur: Two case reports.
The outcome of limb salvage treatment for femoral osteosarcoma with pathological fractures in children is currently unknown. The aim of the present study was to present two cases of patients who received limb salvage surgery with preservation of the epiphysis at the Department of Orthopedics of The General Hospital of Jinan Military Commanding Region (Shandong, China). Between January, 2007 and January, 2013, two pediatric patients were admitted to our hospital with pathological fractures. One of the patients was a girl, aged 11 years, with confirmed osteosarcoma of the right distal femur; the other patient was a boy, aged 9 years, with osteosarcoma of the left distal femur. After receiving two cycles of neoadjuvant chemotherapy following tumor biopsy, the patients received limb salvage surgery with epiphyseal preservation, with wide resection of the tumor and biological reconstruction by allogeneic bone and fibular autograft, followed by 10 cycles of adjuvant chemotherapy. With a mean follow-up of 64 months, there were no postoperative complications, local recurrence or metastasis. The limb function recovered well, although limb shortening was observed. The female patient underwent a second fixation and limb lengthening after epiphyseal closure. Therefore, with effective neoadjuvant chemotherapy, limb salvage surgery with epiphyseal preservation is not contraindicated for pediatric patients with pathological fractures from femoral osteosarcoma. Biological reconstruction by allogeneic bone and vascularized fibular autograft following wide tumor resection is a viable option for such patients, with a good postoperative functional outcome.


Lrrk2 localization in the primate basal ganglia and thalamus: a light and electron microscopic analysis in monkeys.
The Leucine Rich Repeat Kinase-2 (LRRK2) gene is a common mutation target in Parkinson's disease (PD), but the cellular mechanisms by which such mutations underlie the pathophysiology of PD remain poorly understood. Thus, to better characterize the neuronal target sites of LRRK2 mutations in the primate brain, we studied the cellular and ultrastructural localization of Lrrk2 immunoreactivity in the monkey basal ganglia. As previously described, the monkey striatum was the most enriched basal ganglia structure in Lrrk2 labeling. Both projection neurons and parvalbumin-containing GABAergic interneurons displayed Lrrk2 immunoreactivity. At the electron microscopic level, striatal Lrrk2 labeling was associated predominantly with dendritic shafts and subsets of putative glutamatergic axon terminals. At the pallidal level, moderate cellular Lrrk2 immunostaining was found in the external globus pallidus (GPe), while neurons in the internal globus pallidus (GPi) were devoid of Lrrk2 immunoreactivity. Strong labeling was associated with cholinergic neurons in the nucleus basalis of Meynert. Midbrain dopaminergic neurons in the primate substantia nigra pars compacta (SNc) and ventral tegmental area harbored a significant level of Lrrk2 labeling, while neurons in the subthalamic nucleus were lightly immunostained. Most thalamic nuclei were enriched in Lrrk2 immunoreactivity, except for the centromedian nucleus that was completely devoid of labeling. Thus, Lrrk2 protein is widely distributed in the monkey basal ganglia, suggesting that gene mutations in PD may result in multifarious pathophysiological effects that could impact various target sites in the functional circuitry of the primate basal ganglia.


Population density of tropical forest frogs: relation to retreat sites.
The forest frog Eleutherodactylus coqui defends specific sites for retreats and nests in the Luquillo Forest, Puerto Rico. The hypothesis that shortages of nest and retreat sites limit population size was tested by placing 100 bamboo frog houses in plots measuring 100 square meters in areas of high frog density. These new sites were readily adopted by adult frogs. After one year, experimental plots had significantly more nests and frogs of all sizes than did control plots.


Oestradiol enhances tumour regression induced by B7-1/IL-2 adenoviral gene transfer in a murine model of breast cancer.
The majority of breast cancers are oestrogen dependent and although current treatment strategies have improved, approximately 50% of the patients will develop metastasis. New treatments that result in long-term systemic immunity are therefore being developed. We have previously shown that adenoviral gene transfer of B7-1/IL-2 to murine breast cancer induces a high rate of complete tumour regression and systemic immunity. Since oestrogens not only affect breast cancer but also have been shown to modulate immune function and secretion of immune-regulatory cytokines, we explored whether administration of oestradiol altered the immune response induced by an adenoviral vector expressing B7-1/IL-2. An oestrogen-dependent murine breast cancer tumour was used in ovariectomised mice, supplemented either oestradiol or placebo. We report the somewhat unexpected finding that intratumoral injection of adenovirus expressing B7-1/IL-2 induces complete tumour regression in 76% of oestradiol-supplemented mice, while only 18% of the tumours regressed in the oestrogen-depleted group. Cured mice in both groups exhibited a similar CTL response against the tumour antigen. However, intratumoral IFN-gamma levels, 2 days after B7-1/IL-2 injection, were significantly higher in mice treated with oestradiol compared to placebo. This may be one mechanism explaining the higher response rate of tumours in oestradiol-replenished mice.


Bacteriological studies of contamination of hydrophilic contact lenses with Staphylococcus epidermitis.
An attempt was made to contaminate hydrophilic contact lens material with Staphylococcus epidermitis. This study was initiated as a result of some clinical investigations which had revealed the presence of coagulase-negative staphylococcal infection in eyes fitted with this type of contact lens. It was though particularly relevant in view of the known association of some strains of this organism with other plastic prostheses. In vitro cultures proved unsuccessful in demonstrating the ability of this bacterium to metabolize the lens material.


Maternal cigarette smoking during pregnancy and the trajectory of externalizing and internalizing symptoms across childhood: Similarities and differences across parent, teacher, and self reports.
Maternal smoking during pregnancy (MSDP) has been associated with symptoms of externalizing (e.g., hyperactivity) and internalizing (e.g., emotional) disorders in childhood. The present research addresses two new questions about the nature of this relation: (1) Do the associations between MSDP and externalizing and internalizing symptoms vary by who reports the symptoms? and (2) Is MSDP associated with changes in symptomatology across childhood? We address these questions with two cohorts from the Longitudinal Study of Australian Children (LSAC). Parents and teachers completed the Strengths and Difficulties Questionnaire up to six times every two years between child ages 4 and 14 in the older cohort (N = 3841) and up to four times between child ages 4 and 10 in the younger cohort (N = 3714); the study children also completed the same questionnaire up to three times starting at age 10. Across the two cohorts, MSDP was associated with more externalizing symptoms as reported by parents, teachers, and self. MSDP was also associated with increases in externalizing symptoms across childhood when teachers assessed the symptoms but not when parents assessed them. Finally, MSDP was not consistently associated with the average level of internalizing symptoms, but it was associated with increases in these symptoms across childhood. The present research indicates a robust association between MSDP and the average level of externalizing symptoms in childhood regardless of who reports the symptoms. It also indicates that whether MSDP is associated with the trajectory of externalizing symptomatology depends on who reports on the symptoms.


Development of number combination skill in the early school years: when do fingers help?
Children's change over time in frequency of finger use on number combinations was examined in relation to their change in accuracy. Performance was tracked longitudinally over 11 time points, from the beginning of kindergarten (mean age = 5.7 years) to the end of second grade (n= 217). Accuracy in number combinations increased steadily during the time period while frequency of finger use declined. Correlations between finger use and accuracy decreased gradually, ranging from 0.60 in kindergarten to -0.15 at the end of second grade. Low-income children showed linear growth in frequency of finger use while middle-income children slowed down by second grade and even started to decline. Although girls and boys showed similar growth patterns in frequency and accuracy, boys used their fingers less often than girls and were more accurate. The findings indicate that finger use is most adaptive when children are first learning number combinations, but this benefit lessens over time.


Evaluating the optimum number of biopsies to assess histological inflammation in ulcerative colitis: a retrospective cohort study.
The optimal ulcerative colitis biopsy protocol is unclear. To evaluate the number of biopsies required to accurately assess microscopic disease activity in ulcerative colitis METHODS: Biopsies from patients with ≥4 rectosigmoid samples, and clinical and endoscopic data, were retrospectively obtained from a prospective biobank. Histology and endoscopic videos were read blindly. A 4-biopsy Robarts Histopathology Index (RHI) reference score, consisting of the worst item-level ratings from four biopsies, was compared to 1-, 2- and 3-biopsy estimates. Agreement was determined using bivariate errors-in-variable regression analysis (acceptance interval: ±8.25). Endoscopic activity and disease location subgroup analyses were also performed. Forty-six patients had ≥4 rectosigmoid biopsies available (N = 287). The 2-biopsy (tolerance interval: -7.66, 4.79) and 3-biopsy (tolerance interval: -4.86, 3.46) RHI scores demonstrated acceptable agreement with 4-biopsy scores. One-biopsy scores demonstrated unacceptable agreement (tolerance interval: -13.99, 7.78). Mean RHI scores using the 2-, 3- and 4-biopsy approaches were similar (6.1 ± 9.6 P = 0.36; 6.8 ± 10.5, P = 0.7; 7.5 ± 11.2), whereas the 1-biopsy estimate was lower (4.4 ± 8.1, P = 0.06). Histological remission rates were identical for the 2-, 3- and 4-biopsy methods (65.2%, P = 1.0). Subgroup analysis demonstrated that three biopsies were required in patients with endoscopically active disease. Sampling additional colonic locations yielded lower histological remission rates compared to rectosigmoid sampling alone (33.3% vs 61.9%, P = 0.1). A minimum of two - conservatively, three - biopsies are required to reliably assess disease activity in a single colonic segment using the RHI. Further studies are needed of endoscopically active patients and sampling locations. These results have implications for biopsy strategies in clinical trials and practice.


Peripheral Endothelial Cell Count Is a Predictor of Disease Severity in Advanced Fuchs Endothelial Corneal Dystrophy.
In advanced Fuchs endothelial corneal dystrophy (FECD), central endothelial changes do not correlate with disease severity. The peripheral endothelial cell count (ECC) has not been studied as a marker of FECD severity. The goal of this study was to determine the relationship between the peripheral ECC and known clinical markers of FECD in advanced cases. Patients with FECD examined between January 1, 2013, and September 1, 2016, by 1 cornea specialist were identified. Medical records from all previous visits were reviewed to include eyes with high-quality central and peripheral in vivo confocal microscopy images performed on the same day as a clinical evaluation. Endothelial photographs were used to perform manual cell counts centrally and peripherally. Clinical grading of FECD from 1 to 4 was performed at the slit-lamp. We identified 154 eyes of 126 patients that met criteria for inclusion. With higher disease grades, central ECC and peripheral ECC decreased, visual acuity worsened, and central corneal thickness (CCT) increased (all P < 0.05). In patients with advanced disease (defined as either grade 3 or 4, CCT >700, or central ECC <350), the peripheral ECC was the best predictor of disease severity and had the highest number of statistically significant correlations with other clinical markers compared with competing variables. In advanced FECD, severity is best determined by the peripheral ECC compared with the central ECC, visual acuity, clinical disease grade, and CCT. The peripheral ECC should be added to the clinical parameters used to evaluate FECD severity.


Conjoined twins with single heart and liver - a case report.
Twin pregnancy is common but conjoined twins are an extremely rare congenital malformation without any known genetic predisposition. The incidence of conjoined twins is estimated to be around 1 in 250,000 live births with a female predilection. We presented an extremely rare case of abdomino-thoracopagus with single heart and liver. In this paper, the imaging of conjoined twins both antenatally and postnatally is reviewed, in particular taking into consideration recent advances with gray scale and color Doppler ultrasound. This case is of particular interest because of the rarity of abdomino-thoracopagus twins with single heart and liver, in literature.


Cabozantinib in neuroendocrine tumors: tackling drug activity and resistance mechanisms.
Neuroendocrine tumors (NETs) are highly vascularized malignancies in which angiogenesis may entail cell proliferation and survival. Among the emerging compounds with antivascular properties, cabozantinib (CAB) appeared promising. We analyzed the antitumor activity of CAB against NETs utilizing in vitro and in vivo models. For cell cultures, we used BON-1, NCI-H727 and NCI-H720 cell lines. Cell viability was assessed by manual count coupled with quantification of cell death, performed through fluorescence-activated cell sorting analysis as propidium iodide exclusion assay. In addition, we investigated the modulation of the antiapoptotic myeloid cell leukemia 1 protein under CAB exposure, as a putative adaptive pro-survival mechanism, and compared the responses with sunitinib. The activity of CAB was also tested in mouse and zebrafish xenograft tumor models. Cabozantinib showed a dose-dependent and time-dependent effect on cell viability and proliferation in human NET cultures, besides a halting of cell cycle progression for endoduplication, never reported for other tyrosine kinase inhibitors. In a transplantable zebrafish model, CAB drastically inhibited NET-induced angiogenesis and migration of implanted cells through the embryo body. CAB showed encouraging activity in NETs, both in vitro and in vivo models. On this basis, we envisage future research to further investigate along these promising lines.


Functional dissection of a human Dr1-DRAP1 repressor complex.
The heterotetrameric Dr1-DRAP1 transcriptional repressor complex was functionally dissected. Dr1 was found to contain two domains required for repression of transcription. The tethering domain interacts with the TATA box binding protein and directs the repressor complex to the promoter. This tethering domain can be replaced by a domain conferring sequence-specific recognition to the repressor complex. In the absence of the tethering domain, Dr1 interacts with its corepressor DRAP1, but this interaction is not functional. The enhancement of Dr1-mediated repression of transcription by DRAP1 requires the tethering domain. The second domain of Dr1 is the repression domain, which is glutamine-alanine rich. A 65-amino-acid polypeptide containing the repression domain fused to the Ga14 DNA binding domain repressed transcription when directed to TATA-containing and TATA-less promoters. This repression domain was also found to functionally and directly interact with the TATA box binding protein.


Cationic lipophosphoramidates with two disulfide motifs: synthesis, behaviour in reductive media and gene transfection activity.
Lipophosphoramidates have previously been identified as efficient vectors for gene delivery. The incorporation of functional groups that respond to a physiological stimulus is hypothesised to further improve the efficacy of this type of vector and eventually reduce its cytotoxicity. In the present work, we report the effects of the incorporation of two disulfide motifs into the hydrophobic domain, close to the phosphoramidate group. Three cationic vectors possessing such a red/ox sensitive function were synthesised. The capability of one of them (5b) to compact DNA is reported jointly with its ability to release that DNA in the presence of a reducing agent. Finally, compound 5b was tested as a vector for gene delivery into human cells in vitro and its cytotoxicity was also evaluated.


Persistent symptoms among survivors of Hodgkin's disease: an explanatory model based on classical conditioning.
Persistent symptoms of nausea, distress, and vomiting triggered by reminders of cancer treatment were examined among 273 Hodgkin's disease survivors, 1 to 20 years posttreatment. Prevalence rates were high for distress and nausea but low for vomiting. Retrospective report of anticipatory symptoms during treatment was the strongest predictor of persistent symptoms, suggesting that treatment-induced symptoms are less likely to persist if conditioning does not occur initially. Time since treatment was also a significant predictor, with patients more recently treated more likely to experience persistent symptoms. Thus, an explanatory model based on classical conditioning theory successfully predicted presence of persistent symptoms. Symptoms also were associated with ongoing psychological distress, suggesting that quality of life is diminished among survivors with persistent symptoms. Recommendations for prevention and treatment of symptoms are discussed.


Factors influencing happiness of the grandmothers raising grandchildren in rural areas of Northern Thailand.
To study the factors influencing happiness of grandmothers raising grandchildren in the rural areas of Northern Thailand. Cross-sectional survey research was conducted among 400 grandmothers, aged 50-79 years, who raised their grandchildren in the rural areas of Northern Thailand. Participants were selected by cluster sampling. Data were collected through a structured interview from April to July 2009 and analyzed by frequency, percentage, Pearson product moment correlation coefficient, and Multiple regression analysis. Nearly half (46.8%) of grandmothers raising grandchildren had high level of happiness, followed by moderate level (40.4%) and low level (12.8%). The factors, which significantly influenced the happiness of the grandmothers, were self-esteem, social support, and family relationships (p-value < 0.05). In addition, self-esteem, social support, and family relationships could significantly predict happiness of the grandmothers by 48.1%. Self-esteem had the highest predictive power of happiness among grandmothers. The factors influencing happiness of grandmothers raising grandchildren were self-esteem, social support, and family relationships. To promote happiness of grandmothers, responsible organizations should establish activities that enhance the grandmother's self-esteem, provide sufficient social support, and promote good family relationships.


Mono- and bimetallic manganese-carbonyl complexes and clusters bearing imidazol(in)ium-2-dithiocarboxylate ligands.
Five complexes with the generic formula fac-[MnBr(CO)3(S2C·NHC)] were obtained by reacting [MnBr(CO)5] with a set of representative imidazol(in)ium-2-dithiocarboxylate zwitterions. These ligands are the adducts of N-heterocyclic carbenes (NHCs) and carbon disulfide. The mononuclear Mn(i) derivatives were coupled with Na[Mn(CO)5] to afford bimetallic [Mn2(CO)6(S2C·NHC)] clusters. Yet, the most convenient strategy to access these dinuclear Mn(0) products implied a direct carbonyl substitution from the [Mn2(CO)10] dimer. The molecular structures of three monometallic and four bimetallic compounds were elucidated by single crystal X-ray diffraction analysis. In the monometallic complexes, the NHC·CS2 ligands exhibited a bidentate κ2-S,S' coordination mode with an S-C-S bite angle of about 116°. In the dinuclear clusters, the CS2- unit acted as a chelate toward one manganese center and as a pseudoallylic ligand toward the other one. The S-C-S bite angle was reduced to ca. 104°. Thus, the zwitterions displayed a remarkable flexibility, which also permitted a staggered arrangement of the carbonyl groups in the bimetallic systems. Examination of the [small nu, Greek, macron]CO absorption bands on IR spectroscopy helped identify the presence of fac-Mn(CO)3 or Mn2(CO)6 motifs, while the 13C NMR chemical shift of the CS2- moiety was a reliable indicator for monitoring its hapticity. Whereas the dinuclear clusters were air- and moisture-stable crystalline solids, mononuclear halido derivatives displayed only a limited stability under aerobic conditions. Both types of compounds underwent rather unselective, extensive fragmentations in the gas phase, in sharp contrast with the analogous rhenium derivatives that led to clean sequential decarbonylation processes upon collision-induced dissociation.


Applying Quality by Design Concepts to Pharmacy Compounding.
Compounding of medications is an important part of the practice of the pharmacy profession. Because compounded medications do not have U.S. Food and Drug Administration approval, a pharmacist has the responsibility to ensure that compounded medications are of suitable quality, safety, and efficacy. The Federal Government and numerous states have updated their laws and regulations regarding pharmacy compounding as a result of recent quality issues. Compounding pharmacists are expected to follow good preparation prodecures in their compounding practices in much the same way pharmaceutical manufacturers are required to follow Current Good Manufacturing Procedures as detailed in the United States Code of Federal Regulations. Application of Quality by Design concepts to the preparation process for a compounded medication can help in understanding the potential pitfalls and the means to mitigate their impact. The goal is to build quality into the compounding process to ensure that the resultant compounded prescription meets the human or animal patients' requirements.


Acromio-Clavicular Joint Dislocation Types IV to VI: Does the Outcome with the modified Weaver-Dunn Procedure Justify the Treatment?
Introduction: The optimal surgical treatment for acromioclavicular joint (ACJ) injuries remains controversial. The modified Weaver-Dunn (WD) procedure is one of the frequently used techniques. Recently when it was compared with anatomical autogenous tendon graft reconstruction procedures, the results were inferior. However, these anatomical procedures are technically more demanding with small margin of error and they have tendency for postoperative pain because of extra donor site incision. Materials and Methods: Forty patients with type IV to VI ACJ dislocations were treated by modified WD procedure using non-absorbable synthetic suture passed through the base of coracoid process for augmentation of transferred coraco-acromial (CA) ligament. Functional outcome was assessed using the Oxford Shoulder Score, Nottingham Clavicular Score and Visual Analog Score (VAS) at the final follow-up after surgery. Results: The mean pre-operative Oxford Shoulder Score improved from 25.22±2.64 (range 20 to 30) to 44.75±1.99 (range 40 to 48) and mean pre-operative Nottingham Shoulder Score improved from 49.25±4.91 (range 39 to 58) to 87.27±4.39 (range 79 to 96) at last follow-up after surgery with p-value <0.001. Thirty-five (87.5%) patients had excellent outcomes, four (10%) patients had good outcomes and one (2.5%) patient had fair outcome. Thirty-eight (95%) patients had no pain while two (5%) had moderate pain based on VAS score. Conclusion: Modified Weaver-Dunn is a simple well established technique for grade IV to VI ACJ dislocation. We cannot consider this procedure as old and outdated on the argument that the long term functional outcomes are not suboptimal.


Investigating the Osteoinductive Potential of a Decellularized Xenograft Bone Substitute.
Bone grafting is the second most common tissue transplantation procedure worldwide. One of the alternative methods for bone repair under investigation is a tissue-engineered bone substitute. An ideal property of tissue-engineered bone substitutes is osteoinductivity, defined as the ability to stimulate primitive cells to differentiate into a bone-forming lineage. In the current study, we use a decellularization and oxidation protocol to produce a porcine bone scaffold and examine whether it possesses osteoinductive potential and can be used to create a tissue-engineered bone microenvironment. The decellularization protocol was patented by our lab and consists of chemical decellularization and oxidation steps using combinations of deionized water, trypsin, antimicrobials, peracetic acid, and triton-X100. To test if the bone scaffold was a viable host, preosteoblasts were seeded and analyzed for markers of osteogenic differentiation. The osteoinductive potential was observed in vitro with similar osteogenic markers being expressed in preosteoblasts seeded on the scaffolds and demineralized bone matrix. To assess these properties in vivo, scaffolds with and without preosteoblasts preseeded were subcutaneously implanted in mice for 4 weeks. MicroCT scanning revealed 1.6-fold increased bone volume to total volume ratio and 1.4-fold increase in trabecular thickness in scaffolds after implantation. The histological analysis demonstrates new bone formation and blood vessel formation with pentachrome staining demonstrating osteogenesis and angiogenesis, respectively, within the scaffold. Furthermore, CD31+ staining confirmed the endothelial lining of the blood vessels. These results demonstrate that porcine bone maintains its osteoinductive properties after the application of a patented decellularization and oxidation protocol developed in our laboratory. Future work must be performed to definitively prove osteogenesis of human mesenchymal stem cells, biocompatibility in large animal models, and osteoinduction/osseointegration in a relevant clinical model in vivo. The ability to create a functional bone microenvironment using decellularized xenografts will impact regenerative medicine, orthopedic reconstruction, and could be used in the research of multiple diseases.


Circulating microparticles in cardiovascular disease: implications for atherogenesis and atherothrombosis.
The complex and multifactorial nature of atherogenesis and development of atherothrombotic complications involves numerous interactions between various cell types inside the vascular wall (e.g. macrophages and smooth muscle cells) and in the blood (e.g. leukocytes and platelets). One relatively recent advance in this area is the discovery of circulating microparticles and their role in endothelial damage, platelet activation, hypercoagulability and regulation of inter-cellular interactions. Microparticles are small anucleoid phospholipid vesicles released from different cells, such as platelets, erythrocytes, leukocytes and endothelial cells. Microparticles carry surface proteins and include cytoplasmic material of the parental cells responsible for the exertion of microparticle-mediated biological effects. About 25% of the procoagulant activity of stimulated platelet suspensions is associated with microparticles released upon platelet activation and their surface may be approximately 50-100-fold more procoagulant than the surface of activated platelets per se. The available lines of evidence indicate that shedding of microparticles from the parental cells is not just a passive process accompanying cellular dysfunction and apoptosis, but a tightly regulated mechanism implicated in the interactions between various cell types. The role of microparticles as biological messengers is supported by their differential and specific involvement in the pathophysiology of different cardiovascular disorders, including atherogenesis and thrombosis.


Predictors of exercise-induced oxygen desaturation in systemic sclerosis patients with interstitial lung disease.
The diffusion capacity of the lung for carbon monoxide (DLCO) is a good marker of disease severity in patients with idiopathic interstitial pneumonia, and is associated with oxygen saturation; however, little is known about DLCO in systemic sclerosis patients with interstitial lung disease. We studied potential predictors of exercise-induced oxygen desaturation in patients with systemic sclerosis. Data were collected prospectively from 80 of 110 consecutive systemic sclerosis patients with normal oxygen saturation (> 95%) at rest, who could perform the 6-min walk test without physical discomfort, including leg pain. Pulmonary function tests and echocardiography were collected from all subjects. Thirty subjects showed a ≥ 4% decline in oxygen saturation during the 6-min walk test (desaturation group). The other subjects were assigned to the normoxic group. The percent-of-predicted values for FVC, FEV1, total lung capacity, DLCO, and DLCO/alveolar volume were lower, and FEV1/FVC was higher, in the desaturation group. Logistic regression analysis showed the percent-of-predicted DLCO as a highly accurate predictor of exercise-induced oxygen desaturation: the area under the receiver operating characteristic curve was 0.92 (cutoff point 56.3%, sensitivity 0.83, specificity 0.86). Five subjects over the cutoff point of the percent-of-predicted DLCO in the desaturation group could not be distinguished from the normoxic subjects with the lung-volume measurements or right-ventricular systolic pressure. The factor underlying exercise-induced oxygen desaturation appeared to be reduced percent-of-predicted DLCO, which was useful as a predictor in over 80% of the subjects.


Intracellular injection of guanyl nucleotides alters the serotonin-induced increase in potassium conductance in Aplysia neuron R15.
The effects of the adenylate cyclase inhibitor GDP beta S on the response of Aplysia neuron R15 to serotonin (5HT) were investigated. Previous studies have demonstrated that 5HT causes an increase in K+ conductance in R15 and that the response is mediated by cAMP. At concentrations in the micromolar range, GDP beta S inhibits the stimulation of adenylate cyclase by 5HT in particulate fractions from Aplysia ganglia. When micromolar concentrations of GDP beta S are injected into neuron R15, there is no effect on the resting membrane conductance, but the increase in K+ conductance normally elicited by 5HT is completely inhibited. Furthermore, the decrease in inward current normally elicited by dopamine (DA), which does not appear to involve cAMP, is not affected by micromolar concentrations of GDP beta S. In addition, application of 8-benzylthio cAMP to R15 can evoke an increase in K+ conductance even after the injection of GDP beta S, which indicates that events subsequent to the activation of adenylate cyclase are not inhibited by the GDP analogue. In contrast, when millimolar concentrations of GDP beta S are injected into R15, direct effects on membrane conductance are observed and the response of R15 to 5HT is enhanced. Although these effects of high concentrations of GDP beta S are only poorly understood, the results with micromolar concentrations are consistent with the hypothesis that stimulation of adenylate cyclase is necessary for the 5HT-induced increase in K+ conductance in neuron R15.


Functional alpha7-containing nicotinic acetylcholine receptors localize to cell bodies and proximal dendrites in the rat substantia nigra pars reticulata.
The substantia nigra pars reticulata (SNr) is the primary output nucleus for the basal ganglia (BG) in the rat. The SNr is reciprocally connected with the pedunculopontine tegmental nucleus (PPN) in the brainstem, which provides cholinergic innervation to most BG nuclei. The cholinergic input into the BG is considered to be important because PPN activity is altered in Parkinson's disease (PD), a neurological disorder involving the BG, and cholinergic pharmacotherapy is beneficial in alleviating some of its symptoms. In order to better understand the role of cholinergic input to the BG, we examined the effects of nicotinic acetylcholine receptor (nAChR) activation in the GABAergic neurons in slices through juvenile rat SNr. With the aide of subtype selective antagonists, we found that SNr neurons express the alpha7 subtype of nAChRs, the function of which we assessed using the whole cell patch-clamp recording technique. Besides alpha7 nAChRs, GABAergic SNr neurons also contained functional non-alpha7 nAChRs. Using local photolysis of caged carbachol, a broad-spectrum cholinergic agonist, we mapped alpha7 nAChR-mediated currents along the visible extent of filled SNr neurons and found that alpha7 nAChRs can be functionally detected as far as 60 microm away from the soma. Our data are paving the way to a better understanding of the physiological roles of nAChRs in the BG.


[Intraoperative radiotherapy of lung cancer. Report of 31 cases].
Intraoperative radiotherapy (IORT) was done in 31 patients with non-small cell lung cancer from July 1989 to July 1990. Most of them (21/31) were found to have a tumor of stage III. A radiation dose of 5 to 25 Gy was delivered after pulmonary resection to residual tumor or subclinical tumor lesion. The postoperative course was uneventful and no clinical complication developed in all patients. The clinical significance, technique, indications, doses and possible complications of IORT were discussed.


Lessons learned from co-production in public health research: the MAMAH case study involving underserved migrant mothers in the UK.
Research suggests some migrant women are at increased risk of mortality and morbidity in the perinatal period; however, there is a gap in co-produced research to improve care. The UK National Institute for Health and Care Research (NIHR) defines co-production as "an approach in which researchers, practitioners, and members of the public work together, sharing power and responsibility". We summarise learnings from our study, which aimed to co-produce solutions to improve maternity care for migrant women in the UK, by working with women to identify the most important research priorities. We recruited 18 underserved migrant women living in the UK who had given birth in the UK within the past 15 years to create a patient advisory panel. They were recruited via national and local non-governmental organisations and snowball sampling using purposive methods to ensure representation from a range of backgrounds, including those who were refugees, asylum seekers, and undocumented. Underserved was defined as asylum seeking, refugee, undocumented, or low-income mothers (those who were experiencing homelessness or in receipt of welfare support). The women are involved in conceptualisation, analysis, and dissemination of the project. The project is a UK National Institute of Health and Care Research (NIHR) Doctoral Fellowship project lasting 3 years with a variety of research workstreams. The research funding application process began in January, 2021, and the project was funded and began in November, 2022. The research team struggled to access comprehensive training on co-production, particularly in how to counter power dynamics. We appointed a Lead Patient Advisor who manages the relationship between the academics and the patient advisors. Additionally, we reimburse women's time, childcare, and travel. We have found that online meetings are preferable, as women do not need to travel or arrange childcare. We meet our patient advisory panel four times per year. Some women have been directly involved in research such as systematic review screening and qualitative interviewing and have been given research training. Our initial research priorities did not align with those of the women, and this helped us to reshape our work. Women said that having a Lead Patient Advisor made it easier to participate, particularly as some issues are traumatic. To mitigate this, we have offered support resources and debriefing. Using online interpreters has been challenging, and we have recently split into different language groups to maximise engagement. Overall, as researchers, we have learned that taking a truly co-produced approach is time-consuming but has ensured our research prioritises the views of migrant women giving birth in the UK. National Institute for Health and Care Research (NIHR).


Reduced levels of prostaglandin precursors in the blood of atopic patients: defective delta-6-desaturase function as a biochemical basis for atopy.
In the plasma phospholipids of a group of 50 young adults with atopic eczema, there was an elevation of cis-linoleic acid associated with a deficit of gamma-linolenic acid and of the prostaglandin precursors, dihomogammalinolenic acid and arachidonic acid. This suggests that atopics have a deficit in the function of the delta-6-desaturase enzyme which converts linoleic acid to gamma-linolenic acid. Carriers of cystic fibrosis tend to be phenotypically atopic, supporting previous suggestions that in homozygote cystic fibrosis patients the key defect may be in the delta-6-desaturase enzyme. Atopic patients may be exceptionally sensitive to side effects of non-steroidal anti-inflammatory agents. They fail to flush in response to application of niacin compounds to the skin, a reaction mediated by prostaglandins. A deficit of prostaglandin precursors would explain both of these observations. That the observed biochemical deficit plays a causative role in the manifestations of atopy was indicated by the fact that in a double-blind, placebo-controlled crossover trial, gamma-linolenic acid in the form of evening primrose oil (Efamol), partially corrected both the biochemical abnormalities and the clinical state.