Autosomal recessive intellectual disability caused by compound heterozygous variants of the EEF1D gene in a Chinese family.
Intellectual disability is a prevalent neurodevelopmental disorder, with the majority of affected children exhibiting global developmental delay before the age of 5 years. In recent years, certain children have been found to carry homozygous variations of the EEF1D gene, leading to autosomal recessive intellectual disability. However, the pathogenicity of compound heterozygous variations in this gene remains largely unknown. Trio whole-exome sequencing and copy number variation sequencing were done for the genetic etiological diagnosis of a 3-year and 11-month-old Chinese boy who presented with brachycephaly, severe to profound global developmental delay, and hypotonia in the lower limbs. In this case, compound heterozygous variants of the EEF1D gene were found in the child through trio whole-exome sequencing; one was a splice variant (NM_032378.6:c.1905+1G>A) inherited from his father, and the other was a nonsense variant (NM_032378.6:c.676C>T) inherited from his mother. The nonsense variant leads to the production of a premature termination (p.Gln226*). These variations have the ability to explain the clinical phenotypes of the child. Our study expands the variation spectrum and provides compelling evidence for EEF1D as a candidate gene for autosomal recessive intellectual disability. However, due to the deficient number of reported cases, researchers need to further study EEF1D and supplement the clinical phenotypes and treatment measures.


Performance and kinetic evaluation of the anaerobic digestion of two-phase olive mill effluents in reactors with suspended and immobilized biomass.
A lab-scale study was conducted on the mesophilic anaerobic digestion of two-phase olive mill effluents constituted by the mixture of the wash waters derived from the initial cleansing of the olives and those obtained in the washing and purification of virgin olive oil. The digestion was conducted in two continuously stirred tank reactors, one with biomass immobilized on Bentonite (reactor B) and other with suspended biomass used as control (reactor C). The reactors B and C operated satisfactorily between hydraulic retention times of 25.0 and 4.0 days and 25.0 and 5.0 days, respectively. Total chemical oxygen demand (TCOD) efficiencies in the ranges of 88.8-72.1% and 87.9-71.2% were achieved in the reactors with immobilized and suspended biomass, respectively, at organic loading rates of between 0.86 and 5.38 g TCOD/ld and 0.86-4.30 g TCOD/ld, respectively. On comparing both reactors for the same OLRs applied, it was observed that the reactor with support was always more efficient and stable showing higher TCOD, SCOD removal efficiencies and lower VFA/alkalinity ratio values than those found in the control reactor. A mass (TCOD) balance around the reactors allowed the methane yield coefficient, Y(G/S), to be obtained, which gave values of 0.31 and 0.30l CH(4)/g TCOD(removed) for reactors B and C, respectively. The cell maintenance coefficients,k(m), obtained by means of this balance were found to be 0.0024 and 0.0036 g TCOD(removed)/g VSSd, respectively. The volumetric methane production rates correlated with the biodegradable TCOD concentration through an equation of the Michaelis-Menten type for the two reactors studied. This proposed model predicted the behavior of the reactors very accurately showing deviations lower than 10% between the experimental and theoretical values of methane production rates.


Changes in the volume and density of calvarial split bone grafts after alveolar ridge augmentation.
One main problem occurring after bone grafting is resorption, leading to insufficient bone volume and quality, and may subsequently cause dental implant failure. Comparison of graft volume and bone density of iliac crest and calvarial transplants determined by animal studies demonstrates significantly lower resorption of bone grafts harvested from the skull. This paper is the first clinical study evaluating bone volume and density changes of calvarial split bone grafts after alveolar ridge reconstruction. Bone volume and density were determined using CT scans and the software program Dicom Works in a total of 51 calvarial grafts after alveolar ridge augmentation in 15 patients. CT scans were taken in all 15 patients immediately after grafting (T0) and before implantation after a postoperative period of 6 months (T1). In five patients (26 calvarial grafts), a 1-year follow-up was performed (T2). A mean volume reduction of 16.2% at T1 (15 patients) and 19.2% at T2 (five patients) was observed. Bone density was high--about 1000 Hounsfield units--and did not change during the 1-year period. At the time of implantation, 41 transplants were classified as quality 1 bone and 10 as quality 2-3 bone. Grafting area and the technique used for grafting (inlay or onlay graft) did not affect the postoperative bone volume reduction. Generalized osteoporosis did not increase the resorption rate of calvarial transplants. Based on these findings, calvarial split bone grafts are a promising alternative for alveolar ridge reconstruction in dental implantology.


Plasma cortisol levels in Guillain-Barré syndrome.
To assess the relationship between plasma cortisol level and Guillain-Barré syndrome-related complications, notably respiratory failure. One third of patients with Guillain-Barré syndrome develop respiratory failure, which is predicted by few early indicators. Adrenal function has rarely been studied in Guillain-Barré syndrome. Prospective study. Intensive care unit in a teaching hospital. Patients with Guillain-Barré syndrome referred to our unit (n = 102). Plasma cortisol levels were measured before baseline and 60 mins after corticotrophin test in 93 patients with Guillain-Barré syndrome at admission, 16 (17%) of whom were ventilated within 24 hrs from admission, 17 (18%) ventilated after the 24th hr and 60 (65%) never ventilated. Mean plasma cortisol levels at baseline and 60 mins after corticotrophin test were 22.9 +/- 11.3 ng/mL and 45.4 +/- 16.1 ng/mL. At baseline, the plasma cortisol levels were significantly higher in 17 (18%) patients, who developed respiratory failure at least 24 hrs later (28.5 +/- 12.1 ng/mL vs. 20.4 +/- 9.6 ng/mL; p = .003) and dysautonomia (33.1 +/- 14.3 ng/mL vs. 21.4 +/- 10.2 ng/mL, p = .003). When adjusting on only validated clinical predictors (i.e., delay between onset and admission <7 days, inability to lift head and vital capacity <60%), baseline cortisol level was the only independent risk factor for respiratory failure (odds ratio: 2.45 per 10 ng/mL [1.23-4.88 ng/mL], p = .01). Fifty-nine patients underwent electrophysiological testing. When adjusting on a validated electrophysiological model (i.e., peroneal proximal/distal compound muscle action potential ratio and vital capacity), baseline cortisol level remained an independent predictor (odds ratio: 2.50 per 10 ng/mL [1.14-5.51 ng/mL], p = .02). Measurement of baseline plasma cortisol levels can be helpful for early detection of patients with Guillain-Barré syndrome at risk for respiratory failure at least 24 hrs later.


Role of the tyrosine corner motif in the stability of neocarzinostatin.
Although the immunoglobulin-like beta-sandwich fold has no specifically conserved function, some common structural features have been observed, in particular a structural motif, the tyrosine corner. Such a motif was described in neocarzinostatin (NCS), a bacterial protein the structure of which is very similar to that of the immunoglobulin domain. Compared with the other beta-sheet proteins, the NCS 'tyrosine corner' presents non-standard structural features. To investigate the role of this motif in the NCS structure and stability, we studied the properties of a mutant where the H bond interaction had been eliminated by replacing the tyrosine with a phenylalanine. This mutation costs 4.0 kcal/mol showing that the NCS 'tyrosine corner' is involved in protein stability as in the other Greek key proteins. This destabilization is accompanied by remote structural effects, including modification of the binding properties, suggesting an increase in the internal flexibility of the protein. With a view to using this protein for drug targeting, these results along with those obtained previously allow us to define clearly the limitations of the modifications that can be performed on this scaffold.


Advanced cancer genetics in neurosurgical research.
RAPID ADVANCES IN the technology used to study nucleic acids have revealed a great deal regarding the underlying biology of cancer. Most cancers arise as a result of chromosomal rearrangements and deoxyribonucleic acid mutations that lead to the activation of proto-oncogenes and loss of function of tumor suppressor genes. There are a number of different molecular routes that lead to these common goals, necessitating several different techniques of mutational analysis. Although many of these techniques can be difficult in practice, most are conceptually simple. We discuss several of the current techniques in cytogenetics and molecular genetics that are widely used in cancer biology laboratories. Understanding the molecular events that lead to cancer should allow the future development of targeted, nontoxic therapeutics similar to modern-day antibiotics. These technologies are being progressively applied in clinical neurosurgery, where they will be used to detect, diagnose, stratify, and treat cancers of the nervous system. High demand from an increasingly educated patient population means that neurosurgeons will need to be familiar with many of these techniques.


Trauma surgery in the era of nonoperative management: the Dutch model.
Falling operative experience and diminished job satisfaction of trauma surgeons appears to be in part the result of nonoperative management of many blunt injuries. In The Netherlands, the responsibility of trauma surgeons includes the operative treatment of most pelvic and extremity injures, as well as the overall coordination of care. This study describes the type and number of operative procedures performed by a group of trauma surgeons at a trauma center in Amsterdam, The Netherlands. The study was conducted in a university Level I trauma center with 1,250 annual trauma admissions (90% blunt trauma), of which 125 had an Injury Severity Score of 16 or above. During a 3-year period (2001-2003), all operative interventions performed by or under direct supervision of the trauma surgeons were retrospectively analyzed and categorized into eight groups. During the 3-year period, 2,011 operations were performed by the trauma surgeon group. Of these, 1,459 were single procedures and 552 were multiple interventions leading to a total of 2,784 procedures. Nonurgent procedures (n = 915) constituted 45% of the operations, whereas acute procedures (n = 1,096) accounted for 55% of the operations. Of the acute operations, almost 60% were performed during office hours; the remaining operations were performed outside office hours (evening 37%, night 13%, weekend 50%). This study shows that the addition of (non)operative fracture care results in a viable mix of surgical and nonsurgical management. This leads to broad skills and could enhance job satisfaction. These data could be of interest for the current discussion on the future of trauma surgery in North America and might give a lead to increase the attractiveness of our profession for future trauma surgeons.


Impact of ambient bright light on agitation in dementia.
To evaluate the effect of ambient bright light therapy (BLT) on agitation among institutionalized persons with dementia. High intensity, low glare ambient lighting was installed in activity and dining areas of a state psychiatric hospital unit in North Carolina and a dementia-specific residential care facility in Oregon. The study employed a cluster-unit crossover design involving four ambient lighting conditions: AM bright light, PM bright light, All Day bright light, and Standard light. Sixty-six older persons with dementia participated. Outcome measures included direct observation by research personnel and completion by staff caregivers of the 14-item, short form of the Cohen-Mansfield Agitation Inventory (CMAI). Analyses of observational data revealed that for participants with mild/moderate dementia, agitation was higher under AM light (p = 0.003), PM light (p < 0.001), and All Day light (p = 0.001) than Standard light. There was also a trend toward severely demented participants being more agitated during AM light than Standard light (p = 0.053). Analysis of CMAI data identified differing responses by site: the North Carolina site significantly increased agitation under AM light (p = 0.002) and PM light (p = 0.013) compared with All Day light while in Oregon, agitation was higher for All Day light compared to AM light (p = 0.030). In no comparison was agitation significantly lower under any therapeutic condition, in comparison to Standard lighting. Ambient bright light is not effective in reducing agitation in dementia and may exacerbate this behavioral symptom.


Global analysis of proteasomal substrate specificity using positional-scanning libraries of covalent inhibitors.
The proteasome is a large protease complex consisting of multiple catalytic subunits that function simultaneously to digest protein substrates. This complexity has made deciphering the role each subunit plays in the generation of specific protein fragments difficult. Positional scanning libraries of peptide vinyl sulfones were generated in which the amino acid located directly at the site of hydrolysis (P1 residue) was held constant and sequences distal to that residue (P2, P3, and P4 positions) were varied across all natural amino acids (except cysteine and methionine). Binding information for each of the individual catalytic subunits was obtained for each library under a variety of different conditions. The resulting specificity profiles indicated that substrate positions distal to P1 are critical for directing substrates to active subunits in the complex. Furthermore, specificity profiles of IFN-gamma-regulated subunits closely matched those of their noninducible counterparts, suggesting that subunit swapping may modulate substrate processing by a mechanism that does require a change in the primary sequence specificity of individual catalytic subunits in the complex. Finally, specificity profiles were used to design specific inhibitors of a single active site in the complex. These reagents can be used to further establish the role of each subunit in substrate processing by the proteasome.


Discontinuous movement and conformational change during pausing and termination by T7 RNA polymerase.
Time-resolved characterization of T7 RNA polymerase pausing and terminating at a class II termination site has been carried out using site-specifically tethered chemical nucleases. The data indicate that T7RNAP normally moves uniformly down the template as a rigid body. However, at the class II site this movement is interrupted, and the leading edge of the polymerase moves further along the DNA than the trailing edge. This discontinuous movement may persist until it can no longer be accommodated by conformational changes in the elongation complex, at which point the polymerase can either pause or terminate. Termination, but not pausing, is abrogated by introduction of a disulfide bond between the polymerase fingers and thumb subdomains. The introduced cysteines disrupt a thumb-fingers salt-bridge and, under reducing conditions, this mutant enzyme shows reduced processivity coincident with extension of the RNA to 5 nt. These observations suggest that termination requires that the thumb and fingers subdomains move apart, in a reversal of a conformational change important for initially forming a stable transcription complex.


MRP (materiel requirements planning) II: successful implementation the hard way.
Many manufacturing companies embark on MRP II implementation projects as a method for improvement. In spite of an increasing body of knowledge regarding successful implementations, companies continue to attempt new approaches. This article reviews an actual implementation, featuring some of the mistakes made and the efforts required to still achieve "Class A" performance levels.


Temporomandibular symptoms and occlusion: a review of the literature & the concept.
The role of occlusal factors in the etiology of temporomandibular disorders (TMD) is reviewed. The rediscovery of TMD by J.B. Costen in 1934 and the origin of this concept are historically linked. Difficulties in scientifically studying TMD are addressed. TMD lacks specific indicators (signs and symptoms), are episodic and respond to placebos. Clinical success with specific literature (experimental, epidemiological and clinical studies) reviewed fails to support the concept that occlusal factors play a significant role in the etiology of TMD. The remodeling capacity of the temporomandibular joint demonstrates that the joint can accommodate and adapt to various occlusal conditions.


Sol-gel synthesis of Eu3+ doped silica-gold nanorod composites with tunable optical properties.
Gold nanorods (AuNRs) suspension at various concentrations was added into the sol-gel process to engineer nanostructured europium-doped silica host matrices as light-emitting composites. For this purpose, the samples were prepared following two different routes depending on the chemicals used as dopant and catalyst: (a) Eu(NO3)3·5H2O and HNO3, and (b) EuCl·6H2O and HCl. In any case, samples adding various concentrations of AuNRs suspension were prepared. The structural characterization of the samples was through STEM, backscattered electrons (BSE), and EDS analysis. Additionally, their optical properties were evaluated by PL spectroscopy and CIE colorimetry. The results confirmed that (a) methodology produced samples with AuNRs embedded and randomly distributed in the samples. However, these features were not observed in the samples obtained through (b) due to AuNRs dissolution in HCl media. Regarding the optical properties, the analysis of the relative intensity ratio 5D0 → 7F2/5D0 → 7F1 suggested that Eu3+ ions occupy non-centrosymmetric sites in (a) host matrices and centrosymmetric sites in (b). Hence, the increase of AuNRs suspension when fabricating (a) host matrices produced remarkable color changes in the luminescence of the samples towards the reddish-orange region. Meanwhile, the dissolution of AuNRs in (b) minimized the localized surface plasmon resonance (LSPR) effects on the Eu3+ luminescence. These findings revealed that the evaluation and selection of chemicals are critical factors when engineering these materials for more efficient coupling between the LSPR and Eu3+ luminescence.


Population estimate and morphometry of ovarian preantral follicles from three recently recognized squirrel monkey species: a comparative study.
We describe morphological and morphometrical characteristics of preantral ovarian follicles from three recently recognized Saimiri species: S. macrodon, S. cassiquiarensis and S. vanzolinii; the last one a threatened species. Ovaries from four adult monkeys were evaluated: one pair from a pregnant S. macrodon, two ovarian pairs from S. cassiquiarensis females (one of them pregnant), and one left ovary from a senile S. vanzolinii, applying classical histology. Follicular preantral population was quantified and morphology and morphometry of primordial, primary and secondary follicles were evaluated. Follicular preantral population varied among species, being 347,153 in the ovaries of the S. macrodon, 270,342 and 278,376 in the ovaries of both adult non-pregnant and pregnant S. cassiquiarensis females, and 28,149 in the ovary from a senile S. vanzolinii. Most follicles were at primordial or transition stages, except for the senile S. vanzolinii female, which presented the lowest percentages of primordial and transition follicles when compared with primary and secondary ones. Most preantral follicles (>70%) were morphologically normal in the ovaries from all studied S. macrodon and S. cassiquiarensis females, but the ovary of the senile S. vanzolinii female presented a significant decrease in the percentage of normal follicles (primordial: 61%, transition: 52%, primary: 54%, and secondary: 48%). In general, follicular diameter increased significantly from primordial to transition, and subsequently from primary to secondary follicles.


Cellular Response of Human Osteoblasts to Different Presentations of Deproteinized Bovine Bone.
Objectives: This study evaluated the cellular response of primary osteoblasts exposed to two different presentations of a low-temperature non-sintered deproteinized bovine bone matrix (DBBM). Materials and methods: Six different baths of a commercially available DBBM block (Bonefill® Porous Block) and one of DBBM granule (Bonefill® Porous) were evaluated to identify the mineral structure and organic or cellular remnants. Samples of the same baths were processed in TRIZOL for RNA extraction and quantification. For the immunologic cell reaction assay, primary human osteoblasts (pOB) were exposed to DBMM block (pOB + B) or granules (pOB + G), or none (control) for 1, 3, or 7 days of cell cultivation. Expression of proinflammatory cytokines by pOB was evaluated by crosslinked ELISA assay. In addition, total DNA amount, as well as cell viability via LDH evaluation, was assessed. Results: Organic remnants were present in DBBM blocks; 45.55% (±7.12) of osteocytes lacunae presented cellular remnants in blocks compared to 17.31% (±1.31) in granules. In three of five batches of blocks, it was possible to isolate bovine RNA. The highest concentration of TGF-β1 was found in supernatants of pOB + G on day 7 (218.85 ± 234.62 pg/mL) (p < 0.05), whereas pOB + B presented the lowest amount of TGF-β1 secretion at the end of evaluation (30.22 ± 14.94 pg/mL, p < 0.05). For IL-6 and OPG, there was no statistical difference between groups, while pOB + G induced more IL-8 secretion than the control (3.03 ± 3.38 ng/mL, p < 0.05). Considering the kinetics of cytokine release during the study period, all groups presented a similar pattern of cytokines, estimated as an increasing concentration for IL-6, IL-8, and OPG during cultivation. Adherent cells were observed on both material surfaces on day 7, according to H&E and OPN staining. Conclusion: Neither tested material induced a pronounced inflammatory response upon osteoblast cultivation. However, further studies are needed to elucidate the potential influence of organic remnants in bone substitute materials on the regeneration process.


Retrospective analysis of the safety and efficacy of high-dose interleukin-2 after prior tyrosine kinase inhibitor therapy in patients with advanced renal cell carcinoma.
Although tyrosine kinase inhibitors (TKI) are the most common first-line therapy for metastatic renal cell carcinoma, high-dose interleukin-2 (HD-IL2) remains the only agent that provides durable complete responses. The optimal sequence of these agents remains uncertain. This retrospective multi-institutional study examined the safety and efficacy of HD-IL2 following TKI therapy. After IRB approval at 7 HD-IL2 centers, data relating to patient, disease, and treatment characteristics among 40 consecutive patients with metastatic renal cell carcinoma who were treated with HD-IL2 after at least 1 prior TKI therapy were retrospectively collected. The most common cardiac adverse events were grade 3 hypotension and vascular leak syndrome. Six patients (15%) experienced other grade ≥3 cardiac adverse events. There were 2 treatment-related deaths due to congestive heart failure, occurring in 1 patient with short TKI to HD-IL2 interval and another patient with an abnormal baseline cardiac stress test. Best responses included 2 CRs (5%, duration 40+ and 62+ mo), 3 PRs (8%, duration 6, 11, and 24 mo), 13 SD (32%, median duration 12 mo), 20 PD (50%), and 2 not evaluable patients. Median overall survival was 22 months. Administration of HD-IL2 could be safe and effective after TKI therapy; however, careful selection of patients is critical. We recommend baseline cardiac risk factor assessment, screening with both cardiac stress test and echocardiogram, and allowing a TKI to HD-IL2 interval of at least 2 months.


A GSK3-binding peptide from FRAT1 selectively inhibits the GSK3-catalysed phosphorylation of axin and beta-catenin.
The Axin-dependent phosphorylation of beta-catenin catalysed by glycogen synthase kinase-3 (GSK3) is inhibited during embryogenesis. This protects beta-catenin against ubiquitin-dependent proteolysis, leading to its accumulation in the nucleus, where it controls the expression of genes important for development. Frequently rearranged in advanced T-cell lymphomas 1 (FRAT1) is a mammalian homologue of a GSK3-binding protein (GBP), which appears to play a key role in the correct establishment of the dorsal-ventral axis in Xenopus laevis. Here, we demonstrate that FRATtide (a peptide corresponding to residues 188-226 of FRAT1) binds to GSK3 and prevents GSK3 from interacting with Axin. FRATtide also blocks the GSK3-catalysed phosphorylation of Axin and beta-catenin, suggesting a potential mechanism by which GBP could trigger axis formation. In contrast, FRATtide does not suppress GSK3 activity towards other substrates, such as glycogen synthase and eIF2B, whose phosphorylation is independent of Axin but dependent on a 'priming' phosphorylation. This may explain how the essential cellular functions of GSK3 can continue, despite the suppression of beta-catenin phosphorylation.


pH-dependence of the phospholipid interaction of diphtheria-toxin fragments.
Photoreactive phospholipids have been used to probe the lipid interaction of diphtheria toxin. Low pH values induce the membrane insertion of both the binding and enzymic fragments of the toxin. The efficiency of this process is much higher with asolectin than with egg lecithin (phosphatidylcholine)/cholesterol liposomes. The low-pH-induced interaction of the toxin fragments with the membrane hydrocarbon phase is more evident for the enzymic A-chain than for the binding B-chain, and it is fully reversed by returning the pH to neutrality.


Chinese medicine formula lingguizhugan decoction improves Beta-oxidation and metabolism of Fatty Acid in high-fat-diet-induced rat model of Fatty liver disease.
Lingguizhugan decoction (LGZG), a classic traditional Chinese medicine (TCM) formula, has been used to treat obesity and hyperlipidemia in recent years, but the related mechanisms underlying the regulation of lipid metabolism by LGZG are not clear yet. Here, we reported the effectiveness and possible mechanisms of LGZG on rats with fatty liver disease induced by high-fat diet (HFD). Our results demonstrated that LGZG significantly attenuated HFD-induced fatty liver disease, as measured by body weight, liver index, epididymal fat pad-body weight ratio (EFP/BW), liver injury, and hepatic triglycerides (TG) probably through increasing serum thyroid hormone levels, improving beta-oxidation (via modulation of TR β 1 and CPT1A expression), metabolism and transport (through modulation of SREBP-1c, ACSL and ApoB100 expression) of fatty acid. In addition, we discovered the herbal combination with the properties of warming yang to relieve water retention in the formula and proposed the biological basis of LGZG conventional effect via further study on disassembled formula. This study, for the first time, revealed the mechanisms through which LGZG regulates lipid metabolism. Furthermore, our study suggested that it might be feasible to understand the scientific implications of TCM from the perspective of classic formulas' conventional efficacy.


Heavy metal deposition in moss samples from east and south Marmara region, Turkey.
A survey of atmospheric heavy metal deposition in the east and south Marmara region, Turkey was carried out in September 2004. For this purpose, moss samples (Hypnum cupressiforme) were collected in a systematic network of 125 sites. Concentrations of the elements (Al, B, Ba, Ca, Cd, Fe, Co, Cr, Cu, K, Li, Mn, Na, Ni, Pb, Sr, Mg, Ti, and Zn) in the moss were used as an indication of the level of air pollution in the region. Significant differences in heavy metal concentrations, especially for Pb, Cd, Cu, and Zn, were recorded in the moss samples collected around industrialized and heavily populated cities (Istanbul, Bursa, Bandırma, Kocaeli, Biga-Çan) and in an abandoned lead-mining area (Balıkesir-Balya). A map of the spatial distribution of each element in the region was plotted, and enrichment factors were calculated. VARIMAX principal component analysis was applied to the data obtained, and five different components were obtained. The results showed that Pb, Cu, Cd, and Zn derived from anthropogenic and industrial sources while other elements came mostly from natural sources.


Localization of broadband sounds carrying interaural time differences: Effects of frequency, reference location, and interaural coherence.
The auditory processes involved in the localization of sounds in rooms are still poorly understood. The present study investigated the auditory system's across-frequency processing of interaural time differences (ITDs) and the impact of the interaural coherence (IC) of the stimuli in ITD discrimination and localization. First, ITD discrimination thresholds were measured as a function of signal frequency, reference ITD, and IC using critical-band wide noises. The resulting data were fitted with a set of analytical functions and ITD weights were derived using concepts from signal detection theory. Inspired by the weighted-image model [Stern, Zeiberg, and Trahiotis. (1988). J. Acoust. Soc. Am. 84, 156-165], the derived ITD weights were then integrated in a simplified localization model using an optimal combination of ITD information across frequency. To verify this model, a series of localization experiments were conducted using broadband noise in which ITD and IC were varied across frequency. The model predictions were in good agreement with the experimental data, supporting the assumption that the auditory system performs a weighted integration of ITD information across frequency to localize a sound source. The results could be valuable for the design of new paradigms to measure localization in more complex acoustic conditions and may provide constraints for future localization models.


An interlaboratory study of perfluorinated alkyl compound levels in human plasma.
We conducted an interlaboratory study which differed from the typical study of this type because of its emphasis on comparing intralaboratory variability in results. We sent specimens to six laboratories experienced in the analysis of perfluorinated alkyl compounds in blood matrices and that use stringent procedures to control and assure accuracy and precision. Each received an identical set of 60 plasma specimens that were analyzed in six completely independent batches. Split specimens were included so that within- and between-batch coefficients of variation could be calculated. All laboratories used liquid chromatography-tandem mass spectrometry (LC-MS/MS). The concentrations of perfluorooctanesulfonate (PFOS), perfluorooctanoate (PFOA), and perfluorohexanesulfonate (PFHxS) measured in the specimens in general showed a high level of agreement, although in some cases the agreement was only moderate. The average within- and between-batch coefficient of variation for PFOS was 9.1% and 9.3%; for PFOA was 14.5% and 14.5%; and for PFHxS was 14.5% and 17.0%. The recent availability of labeled internal standards, among other advances, has facilitated improvement in the accuracy and precision of the assays. Considering the degree of between-subject variation in levels among people in background-exposed populations, the results indicate that biomarker-based epidemiologic studies of associations with health could have reasonable precision.


Near-death experiences and the temporal lobe.
Many studies in humans suggest that altered temporal lobe functioning, especially functioning in the right temporal lobe, is involved in mystical and religious experiences. We investigated temporal lobe functioning in individuals who reported having transcendental "near-death experiences" during life-threatening events. These individuals were found to have more temporal lobe epileptiform electroencephalographic activity than control subjects and also reported significantly more temporal lobe epileptic symptoms. Contrary to predictions, epileptiform activity was nearly completely lateralized to the left hemisphere. The near-death experience was not associated with dysfunctional stress reactions such as dissociation, posttraumatic stress disorder, and substance abuse, but rather was associated with positive coping styles. Additional analyses revealed that near-death experiencers had altered sleep patterns, specifically, a shorter duration of sleep and delayed REM sleep relative to the control group. These results suggest that altered temporal lobe functioning may be involved in the near-death experience and that individuals who have had such experiences are physiologically distinct from the general population.


Effects of tactile startle on serotonin content of midbrain raphe neurons in rats.
Rats selected to exhibit comparable levels of behavioral reactivity to tactile startle stimuli were used to examine the effects of the startle testing itself on the levels of serotonin (5-HT) in the dorsal and median raphe nuclei and in the hippocampus. A series of 240 air puff stimuli given in 1 h immediately before sacrifice decreased the intracellular 5-HT content, as measured by microspectrofluorimetry, in the median raphe nucleus, without affecting 5-HT levels in the dorsal raphe nucleus. Animals which were housed in the startle chambers but were not given any stimuli were not different from controls. A comparable decrease in 5-HT in the startled animals was found in the hippocampus, which receives its serotonergic innervation from the median raphe nucleus. The results are discussed in the context of the general inverse relationship between startle reactivity and the functional status of the mesolimbic serotonergic pathway, and in the context of the effects of physical stressors on the utilization of brain 5-HT.


Surgical anatomy and histology of the levator palpebrae superioris muscle for blepharoptosis correction.
The detailed knowledge of the architecture of the upper eyelid is very important in numerous upper eyelid corrective surgeries. The article deals with the detailed anatomy of the major components of the upper lid, which are commonly seen in surgical practice. This study was conducted on 19 human cadavers (12 adults and 7 infants) without pathologic changes in the orbital region and eyelids. Anatomic microdissection of the contents of the orbita was performed bilaterally on 12 orbits from 6 unfixed cadavers (3 male and 3 female). Micromorphologic investigations of the orbital tissue were performed on 8 en bloc excised and formalin-fixed orbits of infant cadavers. Specimens were fixed according to the Duvernoy method. An intra-arterial injection of 5% mixture of melt formalin and black ink was administered into the carotid arterial system. Using routine fixation, decalcination, dehydration, illumination, impregnation and molding procedures in paraplast, specimens were prepared for cross-sections. The measurement of the muscle length and diameter in situ in 6 nonfixed cadavers (12 orbits) showed an average length of the levator palpbrae superioris (LPS) muscle body of the 42.0 +/- 1.41 mm on the right, and 40.3 +/- 1.63 mm on the left side. In all the cases, the LPS had blood supply from 4 different arterial systems: the lacrimal, supratrochlear, and supraorbital artery and muscle branches of the ophthalmic artery. The LPS muscle in all the specimens was supplied by the superior medial branch of the oculomotor nerve. The connective tissue associated with the LPS muscle contains two transverse ligaments: the superior (Whitnall's) and intermuscular transverse ligaments (ITL). The orbital septum in all the specimens originated from the arcus marginalis of the frontal bone, and consisted of two layers--the superficial and the inner layer. In addition, a detailed histological analysis revealed that the upper eyelid's crease was formed by the conjoined fascia including the fascia of the orbicularis muscle, the superficial layer of the orbital septum, and the aponeurosis of the LPS muscle, as well as the pretarsal fascia. The conducted study provided a valuable morphological basis for biomechanical and clinical considerations regarding blepharoptosis surgery.


Efficacy of an evidence-based clinical decision support in primary care practices: a randomized clinical trial.
There is consensus that incorporating clinical decision support into electronic health records will improve quality of care, contain costs, and reduce overtreatment, but this potential has yet to be demonstrated in clinical trials. To assess the influence of a customized evidence-based clinical decision support tool on the management of respiratory tract infections and on the effectiveness of integrating evidence at the point of care. In a randomized clinical trial, we implemented 2 well-validated integrated clinical prediction rules, namely, the Walsh rule for streptococcal pharyngitis and the Heckerling rule for pneumonia. INTERVENTIONS AND MAIN OUTCOMES AND MEASURES: The intervention group had access to the integrated clinical prediction rule tool and chose whether to complete risk score calculators, order medications, and generate progress notes to assist with complex decision making at the point of care. The intervention group completed the integrated clinical prediction rule tool in 57.5% of visits. Providers in the intervention group were significantly less likely to order antibiotics than the control group (age-adjusted relative risk, 0.74; 95% CI, 0.60-0.92). The absolute risk of the intervention was 9.2%, and the number needed to treat was 10.8. The intervention group was significantly less likely to order rapid streptococcal tests compared with the control group (relative risk, 0.75; 95% CI, 0.58-0.97; P= .03). The integrated clinical prediction rule process for integrating complex evidence-based clinical decision report tools is of relevant importance for national initiatives, such as Meaningful Use. TRIAL REGISTRATION clinicaltrials.gov Identifier: NCT01386047.


Contralateral effects of upper extremity proprioceptive neuromuscular facilitation patterns.
Electromyography was used to determine the presence of electrical activity in the nonexercised latissimus dorsi, infraspinatus, and pectoralis major muscles while the contralateral limb underwent the proprioceptive neuromuscular facilitation pattern of flexion, abduction, external rotation with elbow straight and extension, adduction, internal rotation with elbow straight. Activity was present in all of these muscles during both components of the pattern. There was no significant difference in activity for the pectoralis major muscle during the flexor as compared to extensor component. The infraspinatus was more active during the flexor component, while the latissimus dorsi was more active during the extensor component. These results could be used in planning a treatment program for patients who are unable to exercise one of their upper extremities and who could benefit from the contralateral effects of upper extremity proprioceptive neuromuscular facilitation patterns.


Characterization of an Immunogenic Mutation in a Patient with Metastatic Triple-Negative Breast Cancer.
Purpose: The administration of autologous tumor-infiltrating lymphocytes (TILs) can mediate durable tumor regressions in patients with melanoma likely based on the recognition of immunogenic somatic mutations expressed by the cancer. There are limited data regarding the immunogenicity of mutations in breast cancer. We sought to identify immunogenic nonsynonymous mutations in a patient with triple-negative breast cancer (TNBC) to identify and isolate mutation-reactive TILs for possible use in adoptive cell transfer.Experimental Design: A TNBC metastasis was resected for TIL generation and whole-exome sequencing. Tandem minigenes or long 25-mer peptides encoding selected mutations were electroporated or pulsed onto autologous antigen-presenting cells, and reactivity of TIL was screened by upregulation of CD137 and IFNγ ELISPOT. The nature of the T-cell response against a unique nonsynonymous mutation was characterized.Results: We identified 72 nonsynonymous mutations from the tumor of a patient with TNBC. CD4+ and HLA-DRB1*1501-restricted TILs isolated from this tumor recognized a single mutation in RBPJ (recombination signal binding protein for immunoglobulin kappa J region). Analysis of 16 metastatic sites revealed that the mutation was ubiquitously present in all samples.Conclusions: Breast cancers can express naturally processed and presented unique nonsynonymous mutations that are recognized by a patient's immune system. TILs recognizing these immunogenic mutations can be isolated from a patient's tumor, suggesting that adoptive cell transfer of mutation-reactive TILs could be a viable treatment option for patients with breast cancer. Clin Cancer Res; 23(15); 4347-53. ©2017 AACR.


Equine cytomegalovirus: structural proteins of virions and nucleocapsids.
Enveloped virions and nucleocapsids of equine cytomegalovirus (ECMV; equine herpesvirus type 2) have been purified from the supernatants and the nuclear extracts of infected rabbit kidney (RK) cells, respectively, and their structural protein compositions have been analyzed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that ECMV nucleocapsids were composed of nine proteins (average molecular weights = 148K, 52K, 49.5K, 46K, 43.5K, 38.5K, 27K, 20K, and 18K), which together constituted 89% of the total nucleocapsid protein on the basis of incorporated 3H-labeled amino acids. The 148K protein comprised 47.3% of the total protein and thus appeared to be similar in molecular weight and proportional composition to the major capsid proteins of other herpesviruses. Purified virions were composed of 37 proteins whose average molecular weights ranged from 14K to greater than 200K. Three intense glycoprotein bands (83K, 78K, and 73.5K) as well as four less intensely labeled glycoproteins were detected in [3H]glucosamine-labeled virion preparations. At least 14 structural proteins were readily detected in extracts of infected cells which had been [35S]methionine labeled late in infection, and 11 of these were immunoprecipitated by rabbit antiserum against purified virions. The protein composition of ECMV differs substantially from those of equine herpesvirus type 1 and type 3 as well as from those of other herpesviruses.


Zieve's Syndrome: An Under-reported Cause of Anemia in Alcoholics.
Anemia is a common finding in alcoholics. It is often multifactorial and caused by a combination of liver dysfunction, ineffective erythropoiesis, and poor nutrition. Zieve's syndrome (ZS) is a clinical syndrome that presents with a triad of jaundice, hemolytic anemia, and hyperlipidemia secondary to alcohol use. Herein, we present a case of a 58-year-old male with a history of liver cirrhosis who presented after a fall due to binge drinking and was found to have severe anemia. Workup was consistent with hemolytic anemia with no source of active bleeding. The patient was managed with supportive treatment and blood transfusions which improved his anemia. However, given his advanced liver disease, he developed encephalopathy and subsequently severe aspiration pneumonia. He died 18 days after admission.


"Ex" marks a spot: the stickiness of dirty work and other removed stigmas.
This theoretical paper proposes that some stigmas, once removed, can continue to incite prejudice toward the formerly marked ("stickiness"), essentially restigmatizing individuals and continuing the stressful experience of being a "dirty person" in others' eyes. The authors focus on dirty work roles (e.g., morticians, exotic dancers) as prototypes of sticky marks that can lead to continued devaluation and, due to legitimizing myths about work, may be especially vulnerable to it. The authors argue that stickiness results from internal attributions made by others about dirty work and is influenced by visibility, onset- and offset-controllability, taint, tenure, and how the work ended. The authors conclude with an analysis of how stickiness might affect vocational patterns of former dirty workers.


Protein targeting and integration signal for the chloroplastic outer envelope membrane.
Most proteins in chloroplasts are encoded by the nuclear genome and synthesized in the cytosol. With the exception of most quter envelope membrane proteins, nuclear-encoded chloroplastic proteins are synthesized with N-terminal extensions that contain the chloroplast targeting information of these proteins. Most outer membrane proteins, however, are synthesized without extensions in the cytosol. Therefore, it is not clear where the chloroplastic outer membrane targeting information resides within these polypeptides. We have analyzed a chloroplastic outer membrane protein, OEP14 (outer envelope membrane protein of 14 kD, previously named OM14), and localized its outer membrane targeting and integration signal to the first 30 amino acids of the protein. This signal consists of a positively charged N-terminal portion followed by a hydrophobic core, bearing resemblance to the signal peptides of proteins targeted to the endoplasmic reticulum. However, a chimeric protein containing this signal fused to a passenger protein did not integrate into the endoplasmic reticulum membrane. Furthermore, membrane topology analysis indicated that the signal inserts into the chloroplastic outer membrane in an orientation opposite to that predicted by the "positive inside" rule.


Functionally relevant and functionally disruptive (epileptic) synchronized oscillations in brain slices.
Focal seizurelike events can be induced in experimental preparations by means of a number of distinct manipulations that differ in synaptic mechanisms. Nevertheless, the form of the seizurelike events can be explained with common principles, including long-lasting excitation of pyramidal cell dendrites and recurrent excitation between pyramidal cells that provides synchronization. One means of induction of seizurelike events, tetanic stimulation, induces a more physiologic type of activity before seizures are elicited, that is, gamma-frequency (> 20 Hz) oscillations. Such oscillations, called 40-Hz oscillations, are believed to be important for cognition in vivo. Experimental gamma oscillations depend critically on synaptic inhibition between interneurons, from interneurons to pyramidal cells, and on a tonic drive to pyramidal cells and interneurons by metabotropic glutamate receptors. The function of gamma oscillations appears to be imposition of a precise temporal structure on the firing patterns of pyramidal cells while still allowing the pyramidal cells to influence each other and be influenced by afferents selectively. We suggest that a relative loss of synaptic inhibition, occurring by any of a number of mechanisms, prevents the occurrence of gamma activity, allows recurrent pyramidal cell-pyramidal cell excitation to predominate, and thereby allows neuronal networks to generate functionally disruptive seizures.


Portal Dissemination of Fusarium graminearum in a Patient with Acute Lymphoblastic Leukemia and Febrile Neutropenia.
We present the case of a man with acute lymphoblastic leukemia and prolonged profound neutropenia, who developed an invasive infection by Fusarium graminearum, acquired via non-cutaneous entry, with gastrointestinal symptoms, sigmoid perforation and liver abscesses due to portal dissemination. The etiologic agent was identified using the 18S-ITS1-5.8S-ITS2-28S rRNA sequence gene, from a liver biopsy. The infection was resolved with surgical drainage and antifungal treatment based on voriconazole. As far as we know, there are no previous reports in the literature of cases of human infection due to Fusarium graminearum.


Axillofemoral bypass. A 2-decade experience reviewed.
A retrospective study of axillofemoral bypass performed at Groote Schuur Hospital over a period of 2 decades was undertaken to analyse the results and to re-examine current criteria for patient selection. During this period 109 patients underwent surgery. The primary indication was critical ischaemia (in 88% of cases). Operative mortality was 6,4%. Graft thrombosis and sepsis were identified as the major reasons for graft failure. Overall graft patency at 3 years was 43% and at 5 years 30%. Successful graft thrombectomy improved the overall cumulative patency rate at 5 years to 43%, and patient survival in the same period was 56%. Modern advances in the assessment of anaesthetic risk together with improved postoperative intensive care have allowed more patients who in the past would have been considered candidates for axillofemoral bypass to undergo direct aortic reconstruction. While the results of axillofemoral bypass are inferior to those of aortic grafts, the technique remains a valuable method of limb salvage for the poor-risk patient and a life-saving treatment for aortic graft sepsis.


[Multiple chemical sensitivity is for real].
Diagnosis of multiple chemical sensitivity is based on symptoms described by the patient. Sometimes the symptom is manifested as flushing of the face and as watering of the eyes and nose. Patients with multiple chemical sensitivity do not sense lower concentrations of scents and smells than do healthy persons, but their olfaction does not adapt to the smell. Because the mechanism of the sensitivity is not properly known, a good treatment does not exist. Some patients are alleviated by antihistamines or nasally sprayed corticosteroids or dilute local anesthetics. The best way is to avoid intense smells.


[Changes in lipid and carbohydrate metabolism in patients with severe sepsis of different etiology].
Different origin (heterogeneity) of sepsis is a key stone in many discussions regarding options for the course and outcome, despite the general rules of development of the pathogenic mechanisms. To compare data of systemic inflammation (CRP, PCT, IL-8, IL-6, IL-4, TNF-alpha) and markers of endothelial dysfunction (NO, lactate, D-dimers), also lipid (cholesterol, triglycerides, HDL, LDL, VLDL) and carbohydrate metabolism between the two groups of patients with severe intra-abdominal infection (n = 109) and severe sepsis of other etiologies (n = 53). We found out a significant difference between the groups in serum levels of the CRP, IL-4 and cholesterol at all stages of the study. During severe abdominal sepsis was accompanied by a significantly higher level of cholesterol, LDL and VLDL, as well as higher values of glycaemia. Patients with sepsis other etiology showed a lighter and more dynamic course of the disease was significantly lower 28-day mortality.


Trends in the measurement of health utilities in published cost-utility analyses.
The Panel on Cost-Effectiveness in Health and Medicine recommended the compilation of a catalog of health state utility weights for use in cost-utility analyses (CUAs), and has given methodological recommendations. This study presents an update, through 2001, to our current registry of utility weights (available at http://www.tufts-nemc.org/cearegistry; previously at http://www.hsph.harvard.edu/cearegistry), and documents recent changes in methods used for utility weight elicitation. We searched the English-language medical literature for original CUAs reporting outcomes as cost per quality-adjusted life-year (QALY). Two trained readers independently audited each article, abstracting data on the health state descriptions, corresponding utility weights, methods of elicitation, and sources of the estimates. The utility elicitation methods from 1998 to 2001 were compared with the methods used to obtain utilities before 1998. We identified 306 CUAs published after 1998, reporting 1210 separate health-related utility estimates, bringing the total in our catalog to 2159 weights. Most frequently, health states pertained to the circulatory system and oncology. Methods varied substantially: 36% of authors used direct elicitation (standard gamble, time trade-off or rating scale), 23% used generic health status instruments (EQ-5D, Health Utilities Index, etc.), and 25% estimated weights based on clinical judgment. Community preferences were used in 27% of the values. Compared with pre-1998, utilities published from 1998 to 2001 were more likely to be elicited using a generic instrument, more likely elicited from community samples, and less likely derived from expert opinion, with no formally employed methodology. Increasingly, analysts conducting CUAs are using generic, preference-weighted instruments, and relying on community-based preferences. Our catalog of utility weights provides a useful reference tool for producers and consumers of CUAs, but also highlights the continued need for improvement in methods and transparency.


Metformin in Gestational Diabetes The Offspring Follow Up (MiGTOFU): Associations between maternal characteristics and size and adiposity of boys and girls at nine years.
Data from 98 women recruited in the Metformin in Gestational Diabetes trial and dual-energy X-ray absorptiometry studies of their children at nine years were analysed to investigate associations between maternal measures during pregnancy and their children's size and adiposity. Mothers of boys (n = 56) and girls (n = 42) had been randomised to metformin or insulin treatment at 30.1 ± 2.8 and 29.3 ± 4.1 weeks gestation, respectively. In boys, fat-free mass indexed to height squared was associated with maternal weight, body mass index, maternal glycaemia and metformin treatment. In boys and girls, fat mass indexed to height squared was associated with maternal glycaemia measures before gestational diabetes treatment.


Influence of Rex and intronic sequences on expression of spliced mRNAs produced by human T cell leukemia virus type I.
Expression of the incompletely spliced HTLV-I mRNAs relies on the viral posttranscriptional activator Rex, whose interaction with the Rex-responsive element (RXRE) overcomes effects of cis-acting repressive sequences (CRSs). Studies based on heterologous reporter plasmids identified an intronic CRS in the 5' LTR and a CRS that overlaps with the RXRE. The present study investigated the effects of these elements in the context of spliced viral mRNAs encoding p21Rex (mRNA 1-3), Tax/Rex (mRNA 1-2-3), and Tof (mRNA 1-2-B). All three mRNAs were inefficiently expressed when transcribed in their mature intronless form, with the p21Rex mRNA showing the weakest expression. In contrast, efficient expression of p21Rex was obtained from a plasmid containing the 5' LTR and 3' portion of the genome that encoded a spliceable RNA. The defective expression of the intronless mRNAs reflected the inhibitory activity of the RXRE and the lack of 5' intronic sequences. Insertion of an intronic 5' LTR segment located upstream of the 5' CRS overcame Rex dependence conferred by the RXRE. The activity of this segment was mapped to the major splice donor and sequences overlapping with, but functionally distinct from, a previously described transcriptional enhancer. The three mRNAs responded differently to Rex and to insertion of the constitutive transport element of simian retrovirus type 1. Taken together, these results suggest that expression of the spliced mRNAs is controlled by the relative influence of positive and negative sequences present on the primary transcript as well as the Rex-RXRE interaction.


Respiratory Responses to Two Voice Interventions for Parkinson's Disease.
The purpose of this study was to examine the respiratory strategies used by persons with Parkinson's disease (PD) to support louder speech in response to two voice interventions. Contrasting interventions were selected to investigate the role of internal and external cue strategies on treatment outcomes. LSVT LOUD, which uses an internal cueing framework, and the SpeechVive prosthesis, which employs an external noise cue to elicit louder speech, were studied. Thirty-four persons with hypophonia secondary to idiopathic PD were assigned to one of three groups: LSVT LOUD (n = 12), SpeechVive (n = 12), or a nontreatment clinical control (n = 10). The LSVT LOUD and SpeechVive participants received 8 weeks of voice intervention. Acoustic and respiratory kinematic data were simultaneously collected at pre-, mid- and posttreatment during a monologue speech sample. Intervention outcomes included sound pressure level (SPL), utterance length, lung volume initiation, lung volume termination, and lung volume excursion. As compared to controls, the LSVT LOUD and SpeechVive participants significantly increased SPL at mid- and posttreatment, thus confirming a positive intervention effect. Treatment-related changes in speech breathing were further identified, including significantly longer utterance lengths (syllables per breath group) at mid- and posttreatment, as compared to pretreatment. The respiratory strategies used to support louder speech varied by group. The LSVT LOUD participants terminated lung volume at significantly lower levels at mid- and posttreatment, as compared to pretreatment. This finding suggests the use of greater expiratory muscle effort by the LSVT LOUD participants to support louder speech. Participants in the SpeechVive group did not significantly alter their respiratory strategies across the intervention period. Single-subject effect sizes highlight the variability in respiratory strategies used across speakers to support louder speech. This study provides emerging evidence to suggest that the LSVT LOUD and SpeechVive therapies elicit different respiratory adjustments in persons with PD. The study highlights the need to consider respiratory function when addressing voice targets in persons with PD.


Disulfide bonds of GM2 synthase homodimers. Antiparallel orientation of the catalytic domains.
GM2 synthase is a homodimer in which the subunits are joined by lumenal domain disulfide bond(s). To define the disulfide bond pattern of this enzyme, we analyzed a soluble form by chemical fragmentation, enzymatic digestion, and mass spectrometry and a full-length form by site-directed mutagenesis. All Cys residues of the lumenal domain of GM2 synthase are disulfide bonded with Cys(429) and Cys(476) forming a disulfide-bonded pair while Cys(80) and Cys(82) are disulfide bonded in combination with Cys(412) and Cys(529). Partial reduction to produce monomers converted Cys(80) and Cys(82) to free thiols while the Cys(429) to Cys(476) disulfide remained intact. CNBr cleavage at amino acid 330 produced a monomer-sized band under nonreducing conditions which was converted upon reduction to a 40-kDa fragment and a 24-kDa myc-positive fragment. Double mutation of Cys(80) and Cys(82) to Ser produced monomers but not dimers. In summary these results demonstrate that Cys(429) and Cys(476) form an intrasubunit disulfide while the intersubunit disulfides formed by both Cys(80) and Cys(82) with Cys(412) and Cys(529) are responsible for formation of the homodimer. This disulfide bond arrangement results in an antiparallel orientation of the catalytic domains of the GM2 synthase homodimer.


Brain abscess due to Xylohypha bantiana.
A case of brain abscess due to Xylohypha bantiana without any obvious predisposing factors and with the uncommon clinical feature of seizures is presented.


Endothelin B receptor deficiency predisposes to pulmonary edema formation via increased lung vascular endothelial cell growth factor expression.
Endothelin (ET) may contribute to pulmonary edema formation, particularly under hypoxic conditions, and decreases in ET-B receptor expression can lead to reduced ET clearance. ET increases vascular endothelial cell growth factor (VEGF) production in vitro, and VEGF overexpression in the lung causes pulmonary edema in vivo. We hypothesized that pulmonary vascular ET-B receptor deficiency leads to increased lung ET, that excess ET increases lung VEGF levels, promoting pulmonary edema formation, and that hypoxia exaggerates these effects. We studied these hypotheses in ET-B receptor-deficient rats. In normoxia, homozygous ET-B-deficient animals had significantly more lung vascular leak than heterozygous or control animals. Hypoxia increased vascular leak regardless of genotype, and hypoxic ET-B-deficient animals leaked more than hypoxic control animals. ET-B-deficient animals had higher lung ET levels in both normoxia and hypoxia. Lung HIF-1alpha and VEGF content was greater in the ET-B-deficient animals in both normoxia and hypoxia, and both HIF-1alpha and VEGF levels were reduced by ET-A receptor antagonism. Both ET-A receptor blockade and VEGF antagonism reduced vascular leak in hypoxic ET-B-deficient animals. We conclude that ET-B receptor-deficient animals display an exaggerated lung vascular protein leak in normoxia, that hypoxia exacerbates that leak, and that this effect is in part attributable to an ET-mediated increase in lung VEGF content.


Endoscopic management of cerebrospinal fluid leaks.
Cerebrospinal fluid (CSF) rhinorrhea is an uncommon but important medical condition. It can result from trauma, intracranial hypertension, or be idiopathic in origin. If left untreated, significant sequelae can result including infectious meningitis. Beta-2 transferrin is a sensitive and specific protein marker for CSF. Various radiographic modalities have been used to localize defects, including computed tomography (CT), magnetic resonance imaging, and CT cisternography. In recent years, surgical management of this condition has evolved significantly, primarily being performed endoscopically. Reconstruction can be performed with fat, free mucosal, or fascial grafts, or with vascularized flaps. The endoscopic surgeon should have a thorough understanding of the pathophysiology and diagnosis of CSF rhinorrhea as well as several surgical options in his or her armamentarium available to treat the patient suffering from CSF rhinorrhea.


Cloning, expression, purification and preliminary X-ray analysis of EstN2, a novel archaeal α/β-hydrolase from Candidatus Nitrososphaera gargensis.
EstN2 is a novel α/β-hydrolase originating from the ammonia-oxidizing thaumarchaeon Candidatus Nitrososphaera gargensis. The genome of the organism was sequenced and genes conferring putative lipolytic activity were amplified and cloned into Escherichia coli as a heterologous host. Through function-based screening, esterase and lipase activity was detected. A recombinant enzyme designated EstN2 was successfully expressed, purified and crystallized. The crystals belonged to space group I2, with one molecule per asymmetric unit, and diffracted X-rays to 1.5 Å resolution.


Double-blind, randomized trial of long-chain polyunsaturated fatty acid supplementation in formula fed to preterm infants.
We tested the hypothesis that balanced addition of long-chain polyunsaturated fatty acid (LCPUFA) to preterm formula during the first weeks of life would confer long-term neurodevelopmental advantage in a double-blind, randomized, controlled trial of preterm formula with and without preformed LCPUFA. The participants were 195 formula-fed preterm infants (birth weight <1750 g, gestation <37 weeks) from 2 UK neonatal units and 88 breast milk-fed infants. Main outcome measures were Bayley Mental Developmental Index (MDI) and Psychomotor Developmental Index (PDI) at 18 months and Knobloch, Passamanick and Sherrard's Developmental Screening Inventory at 9 months' corrected age. Safety outcome measures were anthropometry at 9 and 18 months, tolerance, infection, necrotizing enterocolitis, and death. There were no significant differences in developmental scores between randomized groups, although infants who were fed LCPUFA-supplemented formula showed a nonsignificant 2.6-point (0.25 standard deviation) advantage in MDI and PDI at 18 months, with a greater (nonsignificant) advantage (MDI: 4.5 points; PDI: 5.8 points) in infants below 30 weeks' gestation. LCPUFA-supplemented infants were shorter than control infants at 18 months (difference in length standard deviation score: 0.44; 95% confidence interval: 0.08-0.8). No other significant short- or long-term differences in safety outcomes were observed. Breastfed infants had significantly higher developmental scores at 9 and 18 months than both formula groups and were significantly heavier and longer at 18 months than LCPUFA-supplemented but not control infants. With the dose, duration, and preparation of LCPUFA used, efficacy was not demonstrated, although an advantage in later neurodevelopment cannot be excluded by global tests of development up to 18 months, particularly in infants below 30 weeks' gestation. The surprising effect of LCPUFA-supplemented formula on growth 18 months beyond the intervention period needs to be confirmed in other studies using similar supplementation strategies. Additional follow-up of this cohort is critical at an age when more specific tests of cognitive function are possible.


Direct solid-phase fluoroenzymeimmunoassay of 5 beta-pregnane-3 alpha, 20 alpha-diol-3 alpha-glucuronide in urine.
A competitive solid-phase fluoroenzymeimmunoassay has been developed for rapidly measuring 5 beta-pregnane-3 alpha, 20 alpha-diol-3 alpha-glucuronide (Pd-3G) directly in diluted specimens of pre-breakfast urine. The assay involves use of an antiserum to Pd-3G and enzyme-labeled antigen prepared by chemically linking glucose-6-phosphate dehydrogenase (EC 22.214.171.124) to Pd-3G. Antibody-bound and free antigen are separated by use of a solid-phase double antibody: sheep anti-rabbit gamma-globulin coupled to cellulose particles. The solid phase, isolated by centrifugation, is washed free of labeled antigen and endogenous enzyme interferences. Enzyme activity in the bound fraction is then measured fluorometrically with glucose 6-phosphate as the substrate and generated NADPH as a fluorogenic indicator of enzyme activity. The assay is sufficiently sensitive (30-35 pg per assay tube), specific, and reliable for routine use, and results correlate well (r = 0.98) with those by an established specific radioimmunoassay. Because it is suited to routine use, this assay may be applied to detecting ovulation, assessing the function of corpus luteum, and monitoring early pregnancy.


Logistics, risks, and benefits of automated red blood cell exchange for patients with sickle cell disease.
Red blood cell (RBC) transfusions treat and prevent severe complications of sickle cell disease (SCD) and can be delivered as a simple or exchange transfusion. During an exchange, some of the patient's abnormal hemoglobin (Hb) S (HbS) RBCs are removed. An apheresis device can accomplish an automated RBC exchange, simultaneously removing patient's RBCs while returning other blood components along with normal RBCs. Automated RBC exchange is therefore an isovolemic transfusion that can efficiently decrease HbS RBCs while limiting iron loading and hyperviscosity. However, specialized equipment, trained personnel, appropriate vascular access, and increased RBC exposure are required compared to simple or manual RBC exchange. Therefore, risks and benefits must be balanced to make individualized decisions for patients with SCD who require transfusion.


Localization and expression of phospholipase A2 and polyunsaturated fatty acid profile in the testis tissues of Hu sheep.
The fatty acid content and the localization and expression of phospholipase A2 (PLA2) in the testis of Hu sheep were investigated. A total of 18 six-month-old Hu sheep were divided into small group (S, with left testis weight < 50 g), medium group (M, with left testis weight among 90-110 g), and large group (L, with left testis weight >160 g), which had six individuals each. The expression of PLA2 in testicular tissues of different sizes was analyzed by immunohistochemistry, RT-qPCR, and Western blot. The fatty acid profile was detected by gas chromatography. Immunohistochemical labeling determined that PLA2 protein was expressed in the Leydig and Sertoli cells of testis, and the immunohistochemical average optional density in the S group was significantly greater than the L group (P < 0.05). RT-qPCR and Western blot analysis showed that PLA2 in the S group was greater than that in the L group (P < 0.05). Docosahexaenoic acid, ω-3 polyunsaturated fatty acid (PUFA), and total PUFA content in the testis of the L group were significantly less than those of the S and M groups (P < 0.01). This study showed that PLA2 content in the S group was greater than that in the L group.


Comparing label-free quantitative peptidomics approaches to characterize diurnal variation of peptides in the rat suprachiasmatic nucleus.
Mammalian circadian rhythm is maintained by the suprachiasmatic nucleus (SCN) via an intricate set of neuropeptides and other signaling molecules. In this work, peptidomic analyses from two times of day were examined to characterize variation in SCN peptides using three different label-free quantitation approaches: spectral count, spectra index and SIEVE. Of the 448 identified peptides, 207 peptides were analyzed by two label-free methods, spectral count and spectral index. There were 24 peptides with significant (adjusted p-value < 0.01) differential peptide abundances between daytime and nighttime, including multiple peptides derived from secretogranin II, cocaine and amphetamine regulated transcript, and proprotein convertase subtilisin/kexin type 1 inhibitor. Interestingly, more peptides were analyzable and had significantly different abundances between the two time points using the spectral count and spectral index methods than with a prior analysis using the SIEVE method with the same data. The results of this study reveal the importance of using the appropriate data analysis approaches for label-free relative quantitation of peptides. The detection of significant changes in so rich a set of neuropeptides reflects the dynamic nature of the SCN and the number of influences such as feeding behavior on circadian rhythm. Using spectral count and spectral index, peptide level changes are correlated to time of day, suggesting their key role in circadian function.


The novel subtype of major depressive disorder characterized by somatic symptoms is associated with poor treatment efficacy and prognosis: A data-driven cluster analysis of a prospective cohort in China.
There is not yet a valid and evidence-based system to classify patients with MDD into more homogeneous subtypes based on their clinical features. This study aims to identify symptom-based subtypes of MDD and investigate whether the treatment outcomes of those subtypes would be different. The cohort was established at 12 densely populated cities of China. A total of 1487 patients were enrolled. All participants were 18-65 years old and diagnosed with MDD. Participants were followed up at baseline, weeks 4, 8, and 12, and months 4 and 6. K-means algorithm was used to cluster patients with MDD according to clinical symptoms. The network analysis was adopted to characterize and compare the symptom patterns in the clusters. We also examined the associations between the clusters and the clinical outcomes. The optimal number of the clusters was determined to be 2. Each cluster's maximum Jaccard Co-efficient was calculated to be >0.5 (cluster1 = 0.53, cluster 2 = 0.67). The symptom "depressed mood" and some other affective symptoms were the most prominent in cluster 1. Somatic symptoms, such as weight loss and general somatic symptoms, had the greatest expected influence in cluster 2. Compared with the response rates of the patients in the "somatic cluster", those of the patients in the "affective cluster" were significantly higher (P < 0.05). Patients with MDD might be classified into two symptom-based subtypes featured with affective symptoms or somatic symptoms. The treatment efficacy and prognosis of the subtype featured with somatic symptoms may be worse.


Functionalization of Polydopamine via the Aza-Michael Reaction for Antimicrobial Interfaces.
Polydopamine (pDA) coatings afford tremendous versatility due to their capabilities to provide substrate-independent functionalization with a wide range of amine- and thiol-containing molecules. In this work, we developed a new and facile conjugation approach to the formation of β-amino carbonyl linkages between pDA and acrylate/acrylamide molecules via the aza-Michael reaction. Sulfobetaine acrylamide (SBAA), sulfobetaine methacrylate (SBMA), and poly(ethylene glycol) methacrylate (PEGMA) were employed to graft onto pDA films, giving rise to formation of antifouling coatings. Because of the universal adhesive property of pDA, the coating strategy was applied to different substrates, including TiO2, gold, SiO2, Nitinol alloy, polystyrene, and poly(dimethylsiloxane). The variation of surface chemistry and surface wettability upon pDA modification and subsequent conjugation was monitored with X-ray photoelectron spectroscopy (XPS) and water contact angle measurements. Antifouling properties of coatings were challenged by three common Gram-negative and Gram-positive bacteria. Cytocompatibility of the coatings with NIH-3T3 fibroblasts was accessed using MTT assay. The results showed that pDA coatings grafted with SBAA exhibited superhydrophilicity and excellent fouling resistance likely due to the high chemical reactivity of acrylamide, leading to high grafting density. In addition, dual functional coatings containing passive and active antibacterial components were constructed through the in situ deposition of antimicrobial agent, silver nanoparticles, in pDA, followed by the grafting of SBAA for bacterial repellence. The composite coatings allowed reducing adsorption of E. coli by >95%, while killing attached bacteria by up to 98% upon the releasing of Ag(+) ions as measured by inductively coupled plasma mass spectrometry. Consequently, this work paves a new avenue to the grafting strategy to engineer pDA and to the functional bioinspired antifouling interfaces in a substrate-independent fashion.


Ecological Interventionist Causal Models in Psychosis: Targeting Psychological Mechanisms in Daily Life.
Integrated models of psychotic disorders have posited a number of putative psychological mechanisms that may contribute to the development of psychotic symptoms, but it is only recently that a modest amount of experience sampling research has provided evidence on their role in daily life, outside the research laboratory. A number of methodological challenges remain in evaluating specificity of potential causal links between a given psychological mechanism and psychosis outcomes in a systematic fashion, capitalizing on longitudinal data to investigate temporal ordering. In this article, we argue for testing ecological interventionist causal models that draw on real world and real-time delivered, ecological momentary interventions for generating evidence on several causal criteria (association, time order, and direction/sole plausibility) under real-world conditions, while maximizing generalizability to social contexts and experiences in heterogeneous populations. Specifically, this approach tests whether ecological momentary interventions can (1) modify a putative mechanism and (2) produce changes in the mechanism that lead to sustainable changes in intended psychosis outcomes in individuals' daily lives. Future research using this approach will provide translational evidence on the active ingredients of mobile health and in-person interventions that promote sustained effectiveness of ecological momentary interventions and, thereby, contribute to ongoing efforts that seek to enhance effectiveness of psychological interventions under real-world conditions.


A review of targeted therapy and immune checkpoint inhibitors for metastatic colorectal cancer.
Surgical resection is the cornerstone of treatment for metastatic colorectal cancer (CRC) and offers the best chance at long-term survival. Unfortunately, most patients do not present with resectable metastatic disease and, among patients who do undergo curative-intent resection, many will develop recurrence. In turn, patients require a multi-disciplinary treatment approach with a combination of chemotherapy, surgery, radiation, and/or liver directed therapies that is guided by patient disease burden and clinical status. The development of targeted therapies has led to varying success in other cancers and has emerged as a treatment option for patients with metastatic CRC. While cytotoxic chemotherapy aims to kill cells as they replicate, targeted therapies are directed at biologic features of cancers, like angiogenesis or immune checkpoints. Targeted therapy can facilitate a more treatment tailored approach to the unique genomic alterations of the tumor and hopefully deliver more personalized therapy. We herein provide a systematic review of approved targeted therapies and immune checkpoint inhibitors for metastatic CRC and provide an overview of the current literature.


Xanthomonas maltophilia infection in chronic peritoneal dialysis patients.
Xanthomonas maltophilia infection has only been occasionally reported in patients receiving chronic peritoneal dialysis. We describe four cases of Xanthomonas maltophilia infection associated with chronic peritoneal dialysis. Two patients presented with peritonitis and two with exit site infection. All patients were diabetics, who immediately prior to the study had not received antibiotic therapy. Failure to respond to multiple antibiotic therapy resulted in catheter removal in both patients with peritonitis. In those patients with only exit site infections, dialysis could be continued following antibiotic therapy and catheter replacement in one. Catheter loss in our patients was directly attributed to peritonitis with Xanthomonas maltophilia infection.


[Clinical significance and effects of foreign body embolism during the use of central venous catheters].
An embolism caused by catheters or puncture devices is generally a rare complication during the use of central venous catheters. Possible reasons are either mistakes on the part of the user (e.g. shearing off or tearing of catheters or Seldinger wires) or other accidental causes (e.g. faulty products, material fatigue). As patients are at risk of suffering serious injuries following an embolism, foreign bodies should be removed from the patient's cardio circulatory system as quickly and as completely as possible. Two-dimensional echo-cardiography has proved to be very helpful for diagnostics, while interventional radiology is most effective for the removal of intravascular foreign bodies.


Improved Survival in a Long-Term Rat Model of Sepsis Is Associated With Reduced Mitochondrial Calcium Uptake Despite Increased Energetic Demand.
To investigate the relationship between prognosis, changes in mitochondrial calcium uptake, and bioenergetic status in the heart during sepsis. In vivo and ex vivo controlled experimental studies. University research laboratory. Male adult Wistar rats. Sepsis was induced by intraperitoneal injection of fecal slurry. Sham-operated animals served as controls. Confocal microscopy was used to study functional and bioenergetic parameters in cardiomyocytes isolated after 24-hour sepsis. Electron microscopy was used to characterize structural changes in mitochondria and sarcoplasmic reticulum. The functional response to dobutamine was assessed in vivo by echocardiography. Peak aortic blood flow velocity measured at 24 hours was a good discriminator for 72-hour survival (area under the receiver operator characteristic, 0.84 ± 0.1; p = 0.03) and was used in ex vivo experiments at 24 hours to identify septic animals with good prognosis. Measurements from animals with good prognostic showed 1) a smaller increase in mitochondrial calcium content and in nicotinamide adenine dinucleotide fluorescence following pacing and 2) increased distance between mitochondria and sarcoplasmic reticulum on electron microscopy, and 3) nicotinamide adenine dinucleotide redox potential and adenosine triphosphate/adenosine diphosphate failed to reach a new steady state following pacing, suggesting impaired matching of energy supply and demand. In vivo, good prognosis animals had a blunted response to dobutamine with respect to stroke volume and kinetic energy. In situations of higher energetic demand decreased mitochondrial calcium uptake may constitute an adaptive cellular response that confers a survival advantage in response to sepsis at a cost of decreased oxidative capacity.


Phosphorylation of HuR by Chk2 regulates SIRT1 expression.
The RNA binding protein HuR regulates the stability of many target mRNAs. Here, we report that HuR associated with the 3' untranslated region of the mRNA encoding the longevity and stress-response protein SIRT1, stabilized the SIRT1 mRNA, and increased SIRT1 expression levels. Unexpectedly, oxidative stress triggered the dissociation of the [HuR-SIRT1 mRNA] complex, in turn promoting SIRT1 mRNA decay, reducing SIRT1 abundance, and lowering cell survival. The cell cycle checkpoint kinase Chk2 was activated by H(2)O(2), interacted with HuR, and was predicted to phosphorylate HuR at residues S88, S100, and T118. Mutation of these residues revealed a complex pattern of HuR binding, with S100 appearing to be important for [HuR-SIRT1 mRNA] dissociation after H(2)O(2). Our findings demonstrate that HuR regulates SIRT1 expression, underscore functional links between the two stress-response proteins, and implicate Chk2 in these processes.


Neuropsychological performance and quality of life of 10 year survivors of childhood medulloblastoma.
Survivors of medulloblastoma, the most frequently occurring malignant brain tumor of childhood, suffer neuropsychological damage in the first decade after diagnosis. Cognitive performance, psychosocial functioning and quality of life were assessed in medulloblastoma survivors in the second decade after diagnosis. Ten year survivors were evaluated with a battery of neuropsychological tests, and self-report questionnaires regarding quality of life and emotional functioning. Clinical variables examined included the patient's age at diagnosis, duration since diagnosis, treatment, and complications. Sixteen medulloblastoma survivors [mean age at diagnosis: 7.2 years, range: 1-15 years; 6 males] were tested at a mean age of 22.2 years [range: 13.6-27.9 years]. All survivors had been treated with craniospinal radiation therapy; nine were treated with chemotherapy. Significant impairments were identified in more than 50 of survivors on tests within all neuropsychological domains, including attention, memory, visuospatial abilities, motor functioning, language, and executive functioning. Significant impairments were also identified in all psychosocial domains examined, including employment, ability to drive an automobile, participation in normal education, independent living, and dating history. Most importantly, quality of life scores, reported by both survivors and their caretakers, were in the normal range. Survivors of childhood medulloblastoma frequently suffer severe persistent deficits in a wide-range of neuropsychological functional domains. Nevertheless, survivors and their families do not report impaired quality of life. These severe neuropsychological and psychosocial deficiencies justify further attempts to reduce or delay the use of craniospinal radiation therapy for childhood medulloblastoma.


Introduction of a new model for time-continuous and non-contact investigations of in-vitro thrombolysis under physiological flow conditions.
Thrombolysis is a dynamic and time-dependent process influenced by the haemodynamic conditions. Currently there is no model that allows for time-continuous, non-contact measurements under physiological flow conditions. The aim of this work was to introduce such a model. The model is based on a computer-controlled pump providing variable constant or pulsatile flows in a tube system filled with blood substitute. Clots can be fixed in a custom-built clot carrier within the tube system. The pressure decline at the clot carrier is measured as a novel way to measure lysis of the clot. With different experiments the hydrodynamic properties and reliability of the model were analyzed. Finally, the lysis rate of clots generated from human platelet rich plasma (PRP) was measured during a one hour combined application of diagnostic ultrasound (2 MHz, 0.179 W/cm2) and a thrombolytic agent (rt-PA) as it is commonly used for clinical sonothrombolysis treatments. All hydrodynamic parameters can be adjusted and measured with high accuracy. First experiments with sonothrombolysis demonstrated the feasibility of the model despite low lysis rates. The model allows to adjust accurately all hydrodynamic parameters affecting thrombolysis under physiological flow conditions and for non-contact, time-continuous measurements. Low lysis rates of first sonothrombolysis experiments are primarily attributable to the high stability of the used PRP-clots.


Preliminary appraisal of the reliability and validity of the Colorado State University Feline Acute Pain Scale.
The objective of this study was to determine the inter-rater reliability and convergent validity of the Colorado State University Feline Acute Pain Scale (CSU-FAPS) in a preliminary appraisal of its performance in a clinical teaching setting. Sixty-eight female cats were assessed for pain after ovariohysterectomy. A cohort of 21 cats was examined independently by four raters (two board-certified anesthesiologists and two anesthesia residents) with the CSU-FAPS, and intra-class correlation coefficient (ICC) was used to determine inter-rater reliability. Weighted Cohen's kappa was used to determine inter-rater reliability centered on the 'need to reassess analgesic plan' (dichotomous scale). A separate cohort of 47 cats was evaluated independently by two raters (one board-certified anesthesiologist and one veterinary small animal rotating intern) using the CSU-FAPS and the Glasgow Composite Measure Pain Scale (CMPS-Feline), and Spearman rank-order correlation was determined to assess convergent validity. Reliability was interpreted using Altman's classification as very good, good, moderate, fair and poor. Validity was considered adequate if correlation coefficients were between 0.4 and 0.8. The ICC was 0.61 for anesthesiologists and 0.67 for residents, indicating good reliability. Weighted Cohen's kappa was 0.79 for anesthesiologists and 0.44 for residents, indicating moderate to good reliability. The Spearman rank correlation indicated a statistically significant ( P = 0.0003) positive correlation (0.31; 95% confidence interval 0.14-0.46) between the CSU-FAPS and the CMPS-Feline. The CSU-FAPS showed moderate-to-good inter-rater reliability when used by veterinarians to assess pain level or need to reassess analgesic plan after ovariohysterectomy in cats. The validity fell short of current guidelines for correlation coefficients and further refinement and testing are warranted to improve its performance.


Polyomavirus large T antigen binds independently to multiple, unique regions on the viral genome.
To map the polyomavirus large T antigen binding sites on the viral genome we employed a quantitative immunoassay. Defined, radiolabeled fragments of the viral genome were reacted with crude nuclear extracts prepared from lytically infected mouse 3T6 cells, and the fragments bound by large T antigen were immunoprecipitated with anti-T serum and Formalin-fixed Staphylococcus aureus. The immunoprecipitated DNA was then analyzed by gel electrophoresis and autoradiography. By the use of a variety of restriction endonuclease-generated fragments of wild-type and mutant viral DNAs, the region of high-affinity binding was localized to a 153-base-pair stretch between nucleotides 5292 and 152. At least two independent binding sites lie within this region, one upstream and the other downstream of the Bg/I site at nucleotide 87. One of the binding sites is located within sequences required in cis for DNA replication; the other overlaps the TATA box and cap sites of the early transcription unit. The two sites share a common sequence, A/TGAGGC-N4/5-A/TGAGGC, which may serve as the recognition sequence for large T antigen.


[Mediastioscopic management of pleuropericardial cysts].
We report two cases of paratracheal cystic tumors where no preoperative diagnosis could be established. Mediastinoscopy provided the diagnosis of pleuropericardial cyst. The mediastinoscopic approach allowed partial removal in one case and complete removal in the other.


A bibliometric analysis of toxicology research productivity in Middle Eastern Arab countries during a 10-year period (2003-2012).
Bibliometric studies are increasingly being used for research assessment by involving the application of statistical methods to scientific publications to obtain the bibliographics for each country. The main objective of this study was to analyse the research productivity originating from 13 Middle Eastern Arab (MEA) countries with articles published in toxicology journals. Data from January 1, 2003 till December 31, 2012 were searched for documents with specific words in the toxicology field as a "source title" in any one of the 13 MEA countries. Research productivity was evaluated based on a methodology developed and used in other bibliometric studies. Research productivity was adjusted to the national population and nominal gross domestic product (GDP) per capita. Documents (n = 1,240) were retrieved from 73 international peer-reviewed toxicology journals. The h-index of the retrieved documents was 39. Of the 73 journal titles, 52 (69.9%) have their IF listed in the ISI Journal Citation Reports 2012; 198 documents (16.0%) were published in journals that had no official IF. After adjusting for economy and population power, Egypt (193.6), Palestine (18.1), Kingdom of Saudi Arabia (KSA) (13.0), and Jordan (11.5) had the highest research productivity. Countries with large economies, such as the Kuwait, United Arab Emirates (UAE), and Oman, tended to rank relatively low after adjustment of GDP. The total number of citations at the time of data analysis (August 4, 2013) was 10,991, with a median (interquartile range) of 4 (1-11). MEA collaborated more with countries in the MEA regions (16.7%), especially KSA, Egypt, and UAE, followed by Europe (14.4%), especially with the United Kingdom and Germany. The present data show a promising rise and a good start for toxicology research activity in toxicology journals in the Arab world. Research output is low in some countries, which can be improved by investing in more international and national collaborative research projects in the field of toxicology.


Involvement of 14-3-3 proteins in the second epidermal growth factor-induced wave of Rac1 activation in the process of cell migration.
Immense previous efforts have elucidated the core machinery in cell migration, actin remodeling regulated by Rho family small GTPases including RhoA, Cdc42, and Rac1; however, the spatiotemporal regulation of these molecules remains largely unknown. Here, we report that EGF induces biphasic Rac1 activation in the process of cell migration, and UTKO1, a cell migration inhibitor, inhibits the second EGF-induced wave of Rac1 activation but not the first wave. To address the regulation mechanism and role of the second wave of Rac1 activation, we identified 14-3-3ζ as a target protein of UTKO1 and also showed that UTKO1 abrogated the binding of 14-3-3ζ to Tiam1 that was responsible for the second wave of Rac1 activation, suggesting that the interaction of 14-3-3ζ with Tiam1 is involved in this event. To our knowledge, this is the first report to use a chemical genetic approach to demonstrate the mechanism of temporal activation of Rac1.


Lack of association of the esophageal microbiome in adults with eosinophilic esophagitis compared with non-EoE controls.
Changes in the esophageal microbiome have been reported in children with eosinophilic esophagitis (EoE), but few data exist for adults. We aimed to determine whether the esophageal microbiome differs in adults with and without EoE. In a prospective cohort study, adults undergoing outpatient endoscopy were enrolled as incident EoE cases or non-EoE controls. Clinical, endoscopic, and histologic data were collected. An esophageal biopsy was utilized for microbiome analysis. Bacterial DNA was extracted and the V3-V4 region of the 16S rRNA gene was amplified and sequenced. Analyses were performed comparing microbiome features for cases and controls, and within cases for disease features, with correction for multiple hypothesis testing. A total of 24 incident EoE cases (mean age 40 years; 63% male; 100% white; 97 eos/hpf) and 25 controls (mean age 48, 36% male; 76% white; 1 eos/hpf) were analyzed. Principal coordinate analysis ordination failed to distinguish cases from controls. There were no microbiome differences within EoE cases based on clinical phenotype, presence of atopy, or endoscopic features. Use of proton pump inhibitors (PPIs), however, was significantly associated with 5 taxa including SR1 at the phylum level and Burkholderia at the genus level. There were no significant differences in the esophageal microbiome between newly diagnosed EoE cases and non-EoE controls in adults, or within EoE cases based on clinical features. However, given the strong rationale for the esophageal microbiome in EoE pathogenesis, future studies should explicitly consider the presence of PPIs as a confounding feature.


The structure and interactions of the proline-rich domain of ASPP2.
ASPP2 is a pro-apoptotic protein that stimulates the p53-mediated apoptotic response. The C terminus of ASPP2 contains ankyrin (Ank) repeats and a SH3 domain, which mediate its interactions with numerous partner proteins such as p53, NFkappaB, and Bcl-2. It also contains a proline-rich domain (ASPP2 Pro), whose structure and function are unclear. Here we used biophysical and biochemical methods to study the structure and the interactions of ASPP2 Pro, to gain insight into its biological role. We show, using biophysical and computational methods, that the ASPP2 Pro domain is natively unfolded. We found that the ASPP2 Pro domain interacts with the ASPP2 Ank-SH3 domains, and mapped the interaction sites in both domains. Using a combination of peptide array screening, biophysical and biochemical techniques, we found that ASPP2 Ank-SH3, but not ASPP2 Pro, mediates interactions of ASPP2 with peptides derived from its partner proteins. ASPP2 Pro-Ank-SH3 bound a peptide derived from its partner protein NFkappaB weaker than ASPP2 Ank-SH3 bound this peptide. This suggested that the presence of the proline-rich domain inhibited the interactions mediated by the Ank-SH3 domains. Furthermore, a peptide from ASPP2 Pro competed with a peptide derived from NFkappaB on binding to ASPP2 Ank-SH3. Based on our results, we propose a model in which the interaction between the ASPP2 domains regulates the intermolecular interactions of ASPP2 with its partner proteins.


Diversity of autoantibodies in patients with antimitochondrial antibody and their diagnostic value.
A retrospective analysis of clinical and laboratory features of 102 patients, whose sera contained antibody to mitochondria, showed that primary biliary cirrhosis was diagnosed in 50% of them. Immunofluorescence showed that the sera of the patients with primary biliary cirrhosis all had the M2 antimitochondrial antibody staining pattern. A new staining pattern, designated M2(1), which could be mistaken for the M2 pattern, was not found in any patients with either primary biliary cirrhosis or chronic active hepatitis. Other serological variables such as antibody to mitochondria in IgM class, to multiple nuclear dots, and to the XR antigen, were associated with primary biliary cirrhosis, and taken in association with antimitochondrial antibody of M2 type, contribute to the diagnosis of the disease.


Corticotectal and corticothalamic efferent projections of SIV somatosensory cortex in cat.
Substantial corticotectal (and corticothalamic) projections from the cortex of the anterior ectosylvian sulcus (AES) were demonstrated in the cat using the axonal transport methods of autoradiography and horseradish peroxidase. The corticotectal projection arises nearly exclusively from medium-large pyramidal cells in lamina V. One of the densest projecting areas of the AES is the rostral aspect of its superior bank, where a fourth somatotopic representation (SIV) has recently been demonstrated. It terminates in the intermediate and deep laminae of the superior colliculus, where somatic cells are located. The pathway is bilateral but much heavier ipsilaterally than contralaterally. In contrast to the substantial corticotectal projection from SIV and adjacent tissue, there was no unequivocal evidence for a corticotectal projection from traditional somatosensory cortex SI-SIII. This finding, that somatosensory projections to the cat superior colliculus arise from an area outside of SI-SIII, was unexpected on the basis of what is known about visual corticotectal projections. However, it is consistent with the patterns of other cortical projections that terminate in the intermediate and deep laminae of this structure and with the absence of demonstrable corticotectal influences from SI to SIII in this animal. These data are in contrast to demonstrations by other investigators that there is a corticotectal projection from SI cortex in rodents. Apparently there is a fundamental species difference in the organization of descending somatosensory pathways. A corticothalamic projection of the AES was also observed. This descending projection appeared to form a shell of labeled cells and fibers around the ventrobasal complex, but unequivocal terminal labeling within the ventrobasal complex could not be demonstrated. Dense terminal labeling was apparent in the posterior group of thalamic nuclei (PO) where thalamocortical afferents to the AES originate.