Insertion of adenovirus type 12 DNA in the vicinity of an intracisternal A particle genome in Syrian hamster tumor cells.
In the adenovirus type 12 (Ad12)-induced hamster tumor T1111(2) about 10 Ad12 genome equivalents were integrated at different sites. One of the integrated copies proved unstable and was lost from the cellular genome or rearranged upon passage of the cell line, H1111(2), established from this tumor. This unstable site of junction between the left terminus of Ad12 DNA and hamster DNA and the preinsertion site from BHK21 hamster cells was cloned, sequenced, and analyzed. The junction site showed several peculiarities. At the left terminus of Ad12 DNA, the first 64 nucleotides were deleted. At a distance of 127 nucleotides to the left from this junction site, an internal dispersed fragment of Ad12 DNA comprising nucleotides 1290 to 1361 of the authentic Ad12 DNA sequence was inserted into cellular DNA in an inverted orientation relative to the complete Ad12 genome that was located in its vicinity. The 127-nucleotide sequence between the intact Ad12 genome and the separate 72-base-pair (bp) Ad12 DNA fragment was cellular, but it was not identical to the preinsertion sequence at this location. The sequences flanking the termini of the dispersed 72-bp Ad12 DNA fragment were characterized by direct repeats of 9 or 10 nucleotides. To the left of Ad12 nucleotide 1361 in the separate 72-bp fragment, about 620 cellular nucleotides followed which were identical at the occupied and at the preinsertion sites. It was conceivable that the separate 72-bp Ad12 DNA fragment and the cellular sequence of 127 bp to its right had been transposed en bloc from another unknown location. Abutting the 620 nucleotides of cellular DNA to the left of this block, the 3'-terminal sequence of an endogenous, intracisternal A particle (IAP) genome of hamster cells was detected. The possible significance of the proximity of an IAP sequence to an inserted Ad12 genome with respect to the transformation event, to the instability at this site, or to the transcriptional activity of this region is not known. The 620 bp of cellular DNA between the 72-bp Ad12 DNA fragment and the end of the long terminal repeat of the hamster IAP sequence was apparently of a unique type. Transcriptional activity was not found in the approximate region between nucleotides -620 (to the left) and +350 (to the right) relative to the site of Ad12 DNA insertion, but was found outside these boundaries.


Review of Roy adaptation model-based qualitative research.
The aim of this paper is to examine the role of qualitative research in the development of the Roy adaptation model. An exploration of the findings from qualitative research using Roy's adaptation model from 1995 to 2005 is compared with the findings and recommendations from a previous review over a 25-year period (1970-1995). The usefulness of qualitative methods in furthering nursing theory is highlighted. Findings from both reviews support the assumptions of the model while generating new information and demonstrating the valuable place of qualitative research in promoting nursing science.


Cataract-Causing S93R Mutant Destabilized Structural Conformation of βB1 Crystallin Linking With Aggregates Formation and Cellular Viability.
Cataract, opacity of the eye lens, is the leading cause of visual impairment worldwide. The crucial pathogenic factors that cause cataract are misfolding and aggregation of crystallin protein. βB1-crystallin, which is the most abundant water-soluble protein in mammalian lens, is essential for lens transparency. A previous study identified the missense mutation βB1-S93R being responsible for congenital cataract. However, the exact pathogenic mechanism causing cataract remains unclear. The S93 residue, which is located at the first Greek-key motif of βB1-crystallin, is highly conserved, and its substitution to Arginine severely impaired hydrogen bonds and structural conformation, which were evaluated via Molecular Dynamic Simulation. The βB1-S93R was also found to be prone to aggregation in both human cell lines and Escherichia coli. Then, we isolated the βB1-S93R variant from inclusion bodies by protein renaturation. The βB1-S93R mutation exposed more hydrophobic residues, and the looser structural mutation was prone to aggregation. Furthermore, the S93R mutation reduced the structural stability of βB1-crystallin when incubated at physiological temperature and made it more sensitive to environmental stress, such as UV irradiation or oxidative stress. We also constructed a βB1-S93R cellular model and discovered that βB1-S93R was more sensitive to environmental stress, causing not only aggregate formation but also cellular apoptosis and impaired cellular viability. All of the results indicated that lower solubility and structural stability, sensitivity to environmental stress, vulnerability to aggregation, and impaired cellular viability of βB1-S93R might be involved in cataract development.


Contributions of Disease Severity, Psychosocial Factors, and Cognition to Behavioral Functioning in US Youth Perinatally Exposed to HIV.
Among perinatally HIV-infected (PHIV) and perinatally HIV-exposed, uninfected (PHEU) youth, we evaluated the contributions of home environment, psychosocial, and demographic factors and, among PHIV only, HIV disease severity and antiretroviral treatment (ART), to cognitive functioning (CF) and behavioral functioning (BF). A structural equation modeling (SEM) approach was utilized. Exploratory factor analysis was used to reduce predictor variables to major latent factors. SEMs were developed to measure associations between the latent factors and CF and BF outcomes. Participants included 231 PHIV and 151 PHEU youth (mean age = 10.9 years) enrolled in the PHACS adolescent master protocol. Youth and caregivers completed assessments of CF, BF, psychosocial factors and HIV health. Medical data were also collected. Clusters of predictors were identified, establishing four parsimonious SEMs: child-assessed and caregiver-assessed BF in PHIV and PHEU youth. Among both groups, higher caregiver-child stress predicted worse BF. Caregiver resources and two disease severity variables, late presenter and better past HIV health, were significant predictors of CF in PHIV youth. Higher youth CF was associated with better caregiver-reported BF in both groups. Caregiver resources predicted caregiver-reported BF in PHEU youth, which was mediated via youth CF. Among PHIV youth, better past HIV health and caregiver resources mediated the effects of CF on caregiver-assessed BF. Using SEMs, we found a deleterious impact of caregiver and child stress on BF in both groups and of HIV disease factors on the CF of PHIV youth, reinforcing the importance of early comprehensive intervention to reduce risks for impairment.


[Case of LGMD2A (calpainopathy) clinically presenting as Miyoshi distal myopathy].
We reported a 23-year-old woman with distal myopathy and highly elevated serum creatine kinase (CK) caused by calpainopathy. Although muscle weakness was not evident, a muscle CT scan revealed replacement by adipose tissue in the medial head of the gastrocnemius. The gluteus maximus and biceps femoris were also affected to a lesser degree, but the lateral head of the gastrocnemius was preserved. A histological study of a biopsied specimen of the biceps brachii revealed obvious variation in fiber size and a few necrotic or regenerating fibers. Rimmed vacuoles or lobulated fibers were absent in vacuoles. Although the clinical features suggested Miyoshi's distal myopathy, gene analysis of calpain 3 revealed a c.802-9G > A mutation in intron 5 and a c.1319G > A (p.Arg440Gln) in exon 10. Mini-multiplex Western Blotting (MMW) of the patient's muscle showed no band in calpain 3 (p94) and calpain 3 30 kDa fragments and immunoblotting did not reveal any dysferlin abnormalities. Calpainopathy should be also considered in patients with clinical manifestations of Miyoshi distal myopathy.


Urinary incontinence in the elderly: the brain factor.
Recent research has shown that urge incontinence is common in the elderly and is often combined with reduced bladder sensation. It is associated with cognitive impairment and with underperfusion of the frontal lobes of the cortex. To test for an expected preferential association with particular aspects of cognitive function, 47 incontinent patients (25 men and 22 women, median age 78 y) underwent cognitive testing, 24-hour monitoring of bladder function and videourodynamics. Median CAMCOG score was 72/107. Median urine loss in 24 h was 36 g (range 11-1347 g). 17 patients had urodynamic proof of urge incontinence, 8 with normal and 9 with reduced bladder sensation. Impaired orientation in time was more strongly associated with proven urge incontinence than overall cognitive impairment; it was the only significant predictor (P < 0.00005). Praxis, calculation ability, abstract thinking or recent memory were less closely involved. Thus dysfunction of the frontal cortical lobes and impairment of temporal orientation appear to constitute a "brain factor" underlying geriatric urge incontinence, particularly in combination with reduced bladder sensation.


In Vitro Wear of Glass-Ionomer Containing Restorative Materials.
Advertisements of glass-ionomer-containing restorative materials recommend suitability as load-bearing permanent or semi-permanent restorations. Historically, unacceptably high wear rates limit clinical indications of glass-ionomer-containing restorations in this regard. To compare the in vitro wear of contemporary glass-ionomer-containing dental materials commercially advertised for use in permanent dentition as load-bearing restorations in a chewing simulator. Resin composite was tested as a control. A resin-modified glass ionomer (Ionolux, VOCO gmbH), a high viscosity glass-ionomer hybrid system (Equia Forte HT with Equia Coat, GC America), and a bioactive ionic resin with reactive glass filler (Activa Bioactive Restorative, Pulpdent) were evaluated. Filtek Supreme Ultra (3M ESPE) is a visible light-activated resin composite that served as a control. Standardized flat disk-shaped specimens (n=12/group) were submitted to 500,000 cycles with continuous thermal cycling against steatite antagonists. Volumetric wear was measured at 1000, 10,000, 200,000, and 500,000 cycles. There was a statistically significant difference in mean volumetric wear for Activa Bioactive Restorative (p=0.0081, 95% CI: 0.3973, 0.4982) and Equia Forte HT (p<0.001, 95% CI: 1.2495, 1.8493), but no statistically significant difference in mean volumetric wear for Ionolux (p=0.6653) compared to control. Activa Bioactive Restorative wore approximately 60% less than, and Equia Forte HT twice more than Filtek Supreme Ultra on average, respectively. Compared to a resin composite, contemporary glass-ionomer-containing restorative materials advertised for use as load-bearing restorations display measurably variable in vitro wear rates.


Effects of Mosquito Control Adulticides on Sterile Cochliomyia hominivorax (Diptera: Calliphoridae).
Effects of mosquito control adulticides on sterile screwworm flies, Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae), were investigated via bottle bioassays, outdoor cage tests, and exposure to treated vegetation. In bottle bioassays, 43 μg of permethrin via dilution of Evoluer, 474.56 μg of malathion via dilution of Fyfanon, and 25 μg of naled via dilution of Dibrom Concentrate were used to challenge screwworm flies. Permethrin was more toxic to screwworm flies than was malathion, which was more toxic than naled. On succeeding days, permethrin was still lethal to the flies, whereas malathion and naled were less toxic. During outdoor cage trials, screwworm mortality declined as distance from the spray truck increased. Sterile screwworm flies were killed by lower concentrations of permethrin needed to kill black salt marsh mosquitoes, Aedes taeniorhynchus (Wiedemann) (Diptera: Culicidae). Flies exposed to treated vegetation taken from the path of the spray cloud died more quickly than did flies exposed to leaves taken 5 ft inside the canopy. Fly mortality increased as volume mean diameter of droplets increased. In spite of the toxicity of Evoluer to screwworm flies, aspects of their biology make it unlikely that mosquito control operations would affect released flies.


Intracanalicular Optic Nerve Swelling and Signal Change in Fulminant Untreated Idiopathic Intracranial Hypertension.
The role of the optic canal in the pathogenesis of papilloedema has been under scrutiny recently. Whether a larger canal precedes more severe papilloedema or is the result of bone remodelling from chronically raised pressure across a pressure gradient is not clear. The authors present the magnetic resonance imaging findings of a 29-year-old female with fulminant and untreated idiopathic intracranial hypertension. Imaging showed focal expansion and intrinsic signal changes of the intracanalicular optic nerve. The authors discuss the possibility of either fluid accumulation within the optic nerves from a water hammer effect across blocked optic canals resulting from the steep pressure gradient or opticomalacia (optic nerve softening) from chronic ischaemia.


Relaxation in a glassy binary mixture: comparison of the mode-coupling theory to a Brownian dynamics simulation.
We solved the mode-coupling equations for the Kob-Andersen binary mixture using structure factors calculated from Brownian dynamics simulations of the same system. We found, as was previously observed, that the mode-coupling temperature T(c) inferred from simulations is about two times greater than that predicted by the theory. However, we find that many time-dependent quantities agree reasonably well with the predictions of the mode-coupling theory if they are compared at the same reduced temperature epsilon = (T - T(c))/T(c), and if epsilon is not too small. Specifically, the simulation results for the incoherent intermediate scattering function, the mean square displacement, the relaxation time, and the self-diffusion coefficient agree reasonably well with the predictions of the mode-coupling theory. We find that there are substantial differences for the non-Gaussian parameter. At small reduced temperatures the probabilities of the logarithm of single particle displacements demonstrate that there is hopping-like motion present in the simulations, and this motion is not predicted by the mode-coupling theory. The wave-vector-dependent relaxation time is shown to be qualitatively different from the predictions of the mode-coupling theory for temperatures where hopping-like motion is present.


Sealants and preventive restorations: review of effectiveness and clinical changes for improvement.
Sealants are effective caries-preventive agents to the extent they remain bonded to teeth. Preventive resin restorations (PRR) have a proven record, but are susceptible to failure as the overlying sealant fails. Careful analyses of studies reveal a measurable failure rate of sealants (5-10% per year) that must be addressed. Even under the best of circumstances, sealants fail. Therefore, dentistry (as well as third-party systems) must accept that sealants need vigilant recall and proper preventive maintenance. In addition, it is clear that cost-effective use of sealants will involve selective application on teeth with the greatest caries risk. Caries risk analysis of the patient as well as the tooth is an essential step in the treatment planning process. To improve sealant success, new material advances are suggested. Data from studies using an intermediate layer of dentin bonding agent between etched enamel and sealant show dramatic reduction of failure for sealants, particularly in instances of molars judged difficult to seal due to early stage of eruption.


Traumatic Floating Clavicle: Is This a Rare Injury?
Bipolar clavicle dislocation is thought to be a rare injury pattern. Even experienced orthopaedic surgeons may have not come across this entity during their careers. We report a misdiagnosed case of a 65-year-old male who underwent a motorcycle accident and was surgically treated six months post-injury. This case has been the ground for research since then. We have come to the conclusion that this type of injury is probably not so uncommon as previously thought. Careful evaluation is of immense importance during diagnosis protocol, and practitioners should be aware of this injury pattern in order to avoid misdiagnosis.


Novelties in diagnostics and therapy of antibody mediated rejection.
Antibody mediated rejection (AMR) is becoming an increasing challenge in renal allotransplantation. AMR is usually associated with profound allograft dysfunction and inferior allograft survival. In the era of refined crossmatch-techniques and improved solid-phase assays, hyperacute rejections in presensitized patients are less common. Majority of AMRs occur due to de novo production of donor specific antibodies. AMR appears to stand for a spectrum of histologic changes paralled with immunohistochemical positivity for C4d along peritubular capillaries. Removal of antibodies with plasmapheresis or immunoadsorption in combination with neutralizing and immunomodulatory intravenous immunoglobulin are therapy standards in majority of transplant centers worldwide. The addition of anti-CD20 (rituximab) with the aim to reduce the number of B-cells may offer advantage in some cases. Apart from donor specific HLA-antibodies, non-HLA antibodies reacting to arterial antigens have been speculated to be responsible for rejections in some patients. More precise definition of antigen targets for non-HLA antibodies is emerging. The analysis of the subtle mechanistic and diagnostic differences among rejection subtypes will help to identify patients at particular risk and improve outcome of AMR.


Sustainability of cement kiln co-processing of wastes in India: a pilot study.
Co-processing in cement kiln achieves effective utilization of the material and energy value present in the wastes, thereby conserving the natural resources by reducing the use of virgin material. In India, a number of multifolded initiatives have been taken that take into account the potential and volume of waste generation. This paper studies the factors which might influence the sustainability of co-processing of waste in cement kilns as a business model, considering the issues and challenges in the supply chain framework in India in view of the four canonical pillars of sustainability. A pilot study on co-processing was carried out in one of the cement plant in India to evaluate the environmental performance, economical performance, operational performance and social performance. The findings will help India and other developing countries to introduce effective supply chain management for co-processing while addressing the issues and challenges during co-processing of different waste streams in the cement kilns.


Food allergy to pumpkinseed--characterization of allergens.
In recent years, pumpkinseed has become increasingly popular as a foodstuff. Here we report the occurrence of allergic reactions (itching and swelling of oral mucosa, and asthma) to this member of the Cucurbitaceae family. We investigated three patients suffering from symptoms after ingestion of roasted pumpkinseed. All the patients fished for sport and used pressed pumpkinseed flour as bait. Sera were tested by the immunoblot technique for IgE reactivity with proteins of pumpkinseed extract. The immunoblot revealed pumpkinseed allergens of 13, 14, 36, 48, 77, and 87 kDa. Inhibition experiments with recombinant birch profilin were performed: IgE binding to the 14-kDa allergen was completely blocked by preincubation of the sera with recombinant birch profilin. In conclusion, type I allergy to pumpkinseed is rare, and the patients' histories suggest inhalation of pumpkinseed flour during fishing to be the relevant route of sensitization, leading to food allergy to pumpkinseed.


[Plantago ovata].
Plantago ovata is a high fibre bulk forming laxative. It absorbs water and expands to provide increased bulk and moisture content to the stool. The increased bulk encourages normal peristalsis and bowel motility. Clinical Indications: Constipation, Fecal Incontinence, Hemorrhoids, Ulcerative Colitis, Appetite, Hyperlipidemia, Diabetes mellitus.


Phytomedicines (medicines derived from plants) for sickle cell disease.
Sickle cell disease (SCD) is a common recessively inherited disorder of haemoglobin affecting peoples originating from sub-Saharan Africa, the Middle East and Mediterranean basin, the Indian subcontinent, the Caribbean and South America. The homozygous state (SS) is associated with complications and a reduced life expectancy. Phytomedicines (medicine derived from plants in their original state) encompass much of what the populations most affected would encounter in terms of plant-remedies from traditional healers. There has been little in the way of systematic appraisal of their benefits. To assess the benefits and risks of phytomedicines in people with SCD of all types, of any age, in any setting. We searched the Cochrane Cystic Fibrosis and Genetic Disorders group specialised register of controlled trials of haemoglobinopathies, which comprises references identified from comprehensive electronic database searches, handsearches of relevant journals and abstract books of conference proceedings. We performed an additional search of the bibliographic database of Allied and Complementary Medicine (AMED). Date of most recent search of the trials register: September 2003. All randomised or quasi-randomised trials with participants of all ages with SCD, in all settings, comparing the administration of phytomedicines, by any mode to placebo or standard treatment, including blood transfusion and hydroxyurea. Both reviewers independently assessed trial quality and extracted data from the study. Reports of two trials were found, of which only one, including 82 participants, was eligible for inclusion in this review. This Phase IIB (pivotal) study suggests that a phytomedicine, NIPRISAN, was effective in reducing episodes of SCD crisis associated with severe pain over a six-month period. NIPRISAN did not appear to affect the risk of severe complications or the level of anaemia. No serious adverse effects were reported. While NIPRISAN, as a phytomedicine, appeared to be safe and effective, over a six-month follow-up period of this study, in reducing crises associated with severe pain, further studies are required to assess its role in the management of people with sickle cell disease. The results of Phase III, multicentre trials are awaited.


A four-country investigation of factors facilitating student internet use.
Education and national culture may influence Internet use. The present study explored factors that might facilitate college students' use of the Internet across samples in four countries: India, Mauritius, Reunion Island (an overseas Department of France), and the United States. Questionnaire data was collected on students' extent and frequency of Internet use. Also, structured items asked about the factors that would help them to use the Internet more. Data was analyzed using dummy-coded multiple regression so as to compare responses across the country groups. The U.S. sample was coded as the reference group so that the regression coefficients in the equation provided a comparison between the U.S. sample and each other country represented. Further country comparisons were made using t-tests for differences between the regression coefficients. Results suggested that there was more Internet use in the U.S. sample compared to the others. The Non-Western samples also reported greater desire for Internet access and non-English web sites. Training on how to use the Internet and course requirements for Internet use were reported as helpful by the India and Mauritius groups to a greater extent than by those in Reunion and the United States. Implications of the present exploratory results are discussed for applications of theoretical models across cultures.


Pulsed Direct Current Electrospray: Enabling Systematic Analysis of Small Volume Sample by Boosting Sample Economy.
We had developed pulsed direct current electrospray ionization mass spectrometry (pulsed-dc-ESI-MS) for systematically profiling and determining components in small volume sample. Pulsed-dc-ESI utilized constant high voltage to induce the generation of single polarity pulsed electrospray remotely. This method had significantly boosted the sample economy, so as to obtain several minutes MS signal duration from merely picoliter volume sample. The elongated MS signal duration enable us to collect abundant MS(2) information on interested components in a small volume sample for systematical analysis. This method had been successfully applied for single cell metabolomics analysis. We had obtained 2-D profile of metabolites (including exact mass and MS(2) data) from single plant and mammalian cell, concerning 1034 components and 656 components for Allium cepa and HeLa cells, respectively. Further identification had found 162 compounds and 28 different modification groups of 141 saccharides in a single Allium cepa cell, indicating pulsed-dc-ESI a powerful tool for small volume sample systematical analysis.


Cross-linguistic influence of first language writing systems on brain responses to second language word reading in late bilinguals.
Introduction How human brains acquire second languages (L2) is one of the fundamental questions in neuroscience and language science. However, it is unclear whether the first language (L1) has a cross-linguistic influence on the processing of L2. Methods Here, we used functional magnetic resonance imaging to compare brain activities during L2 word reading tasks of phonographic Japanese Kana between two groups of learners of the Japanese language as their L2 and who had different orthographic backgrounds of their L1. For Chinese learners, a L1 of the Chinese language (Hanji) and a L2 of the Japanese Kana differed orthographically, whereas for Korean learners, a L1 of Korean Hangul and a L2 of Japanese Kana were similar. Results Our analysis revealed that, although proficiency and the age of acquisition did not differ between the two groups, Chinese learners showed greater activation of the left middle frontal gyrus than Korean learners during L2 word reading. Conclusion Our results provide evidence that strongly supported the hypothesis that cross-linguistic variations in orthography between L1 and L2 induce differential brain activation during L2 word reading, which has been proposed previously.


NAADP/SERCA3-Dependent Ca2+ Stores Pathway Specifically Controls Early Autocrine ADP Secretion Potentiating Platelet Activation.
Ca2+ signaling is a key and ubiquitous actor of cell organization and its modulation controls many cellular responses. SERCAs (sarco-endoplasmic reticulum Ca2+-ATPases) pump Ca2+ into internal stores that play a major role in the cytosolic Ca2+ concentration rise upon cell activation. Platelets exhibit 2 types of SERCAs, SERCA2b and SERCA3 (SERCA3 deficient mice), which may exert specific roles, yet ill-defined. We have recently shown that Ca2+ mobilization from SERCA3-dependent stores was required for full platelet activation in weak stimulation conditions. To uncover the signaling mechanisms associated with Ca2+ mobilization from SERCA3-dependent stores leading to ADP secretion. Using platelets from wild-type or Serca3-deficient mice, we demonstrated that an early (within 5-10 s following stimulation) secretion of ADP specifically dependent on SERCA3 stored Ca2+ is exclusively mobilized by nicotinic acid adenosine dinucleotide-phosphate (NAADP): both Ca2+ mobilization from SERCA3-dependent stores and primary ADP secretion are blocked by the NAADP receptor antagonist Ned-19, and reciprocally both are stimulated by permeant NAADP. In contrast, Ca2+ mobilization from SERCA3-dependent stores and primary ADP secretion were unaffected by inhibition of the production of IP3 (inositol-1,4,5-trisphosphate) by phospholipase-C and accordingly were not stimulated by permeant IP3. Upon activation, an NAADP/SERCA3 Ca2+ mobilization pathway initiates an early ADP secretion, potentiating platelet activation, and a secondary wave of ADP secretion driven by both an IP3/SERCA2b-dependent Ca2+ stores pathway and the NAADP/SERCA3 pathway. This does not exclude that Ca2+ mobilized from SERCA3 stores may also enhance platelet global reactivity to agonists. Because of its modulating effect on platelet activation, this NAADP-SERCA3 pathway may be a relevant target for anti-thrombotic therapy. Graphic Abstract: A graphic abstract is available for this article.


Genome-wide maps of mononucleosomes and dinucleosomes containing hyperacetylated histones of Aspergillus fumigatus.
It is suggested that histone modifications and/or histone variants influence the nucleosomal DNA length. We sequenced both ends of mononucleosomal and dinucleosomal DNA fragments of the filamentous fungus Aspergillus fumigatus, after treatment with the histone deacetylase inhibitor trichostatin A (TSA). After mapping the DNA fragments to the genome, we identified >7 million mononucleosome positions and >7 million dinucleosome positions. We showed that the distributions of the lengths of the mononucleosomal DNA fragments after 15-min and 30-min treatments with micrococcal nuclease (MNase) showed a single peak at 168 nt and 160 nt, respectively. The distributions of the lengths of the dinucleosomal DNA fragments after 15-min- and 30-min-treatment with MNase showed a single peak at 321 nt and 306 nt, respectively. The nucleosomal DNA fragments obtained from the TSA-treated cells were significantly longer than those obtained from the untreated cells. On the other hand, most of the genes did not undergo any change after treatment. Between the TSA-treated and untreated cells, only 77 genes had >or=2-fold change in expression levels. In addition, our results showed that the locations where mononucleosomes were frequently detected were conserved between the TSA-treated cells and untreated cells in the gene promoters (lower density of the nucleosomes). However, these locations were less conserved in the bodies (higher density of the nucleosomes) of genes with >or=2-fold changes. Our findings indicate that TSA influences the nucleosome positions, especially of the regions with high density of the nucleosomes by elongation of the nucleosomal DNA. However, most of the nucleosome positions are conserved in the gene promoters, even after treatment with TSA, because of the low density of nucleosomes in the gene promoters.


A systematic review of chiropractic management of adults with Whiplash-Associated Disorders: recommendations for advancing evidence-based practice and research.
The literature relevant to the treatment of Whiplash-Associated Disorders (WAD) is extensive and heterogeneous. A Participatory Action Research (PAR) approach was used to engage a chiropractic community of practice and stakeholders in a systematic review to address a general question: 'Does chiropractic management of WAD clients have an effect on improving health status?' A systematic review of the empirical studies relevant to WAD interventions was conducted followed by a review of the evidence. The initial search identified 1,155 articles. Ninety-two of the articles were retrieved, and 27 articles consistent with specific criteria of WAD intervention were analyzed in-depth. The best evidence supporting the chiropractic management of clients with WAD is reported. Further review identified ways to overcome gaps needed to inform clinical practice and culminated in the development of a proposed care model: the WAD-Plus Model. There is a baseline of evidence that suggests chiropractic care improves cervical range of motion (cROM) and pain in the management of WAD. However, the level of this evidence relevant to clinical practice remains low or draws on clinical consensus at this time. The WAD-Plus Model has implications for use by chiropractors and interdisciplinary professionals in the assessment and management of acute, subacute and chronic pain due to WAD. Furthermore, the WAD-Plus Model can be used in the future study of interventions and outcomes to advance evidence-based care in the management of WAD.


Effects of pre-existing hearing loss on proposed ANSI S12.13 outcomes for characterizing hearing conservation program effectiveness: follow-up investigation.
Draft ANSI S12.13 1991 Standard Evaluating the Effectiveness of Hearing Conservation Programs presents statistical protocols purportedly sensitive to temporary threshold shift (TTS) in occupational hearing conservation program (HCP) audiometric databases. Because it is well established that less TTS is found in humans with hearing loss than in those with essentially normal hearing, one might predict baseline hearing level to be a confounding variable in interpreting ANSI S12.13 outcomes, with better-hearing industrial populations tending to demonstrate greater year-to-year audiometric variability than poorer-hearing groups. However, in a large industrial sample from the public domain audiometric database, poorer-hearing groups systematically demonstrated greater audiometric threshold variability than better-hearing groups, and ANSI S12.13 outcome magnitudes were highly positively correlated to baseline hearing levels. These findings generally do not support notions that ANSI S12.13 outcomes clearly provide indirect measures of TTS. HCP managers should exercise extreme caution in interpreting ANSI S12.13 outcomes to rate overall program performance. In particular, HCP managers should not rely upon these outcomes for decisions regarding hearing protection policies without careful consideration of pre-existing hearing loss in the populations involved.


Photo-Responsive Soft Ionic Crystals: Ion-Pairing Assemblies of Azobenzene Carboxylates.
This report delineates the design and synthesis of negatively charged azobenzene derivatives that form photo-responsive ion-pairing assemblies. The azobenzene carboxylates possessing aliphatic chains were prepared as photo-responsive anions that promote the formation of ion-pairing dimension-controlled assemblies, including mesophases, when used in conjunction with a tetrabutylammonium (TBA) cation. The photo-responsive properties of the ion pairs and the precursory carboxylic acids in the bulk state were examined by polarized optical microscopy (POM) and X-ray diffraction (XRD), demonstrating that liquid crystal (LC)-liquid and crystal-liquid phase transitions occurred, depending on the number and lengths of the aliphatic chains of each assembly. An ion pair exhibited photo-induced crystal-crystal phase transitions upon switching between two irradiation wavelengths (365/436 nm).


Immunoscintigraphy of colon carcinoma.
Two I-131 labeled monoclonal antibodies that react specifically with human gastrointestinal cancers in cell cultures were administered to 90 cancer patients for the scintigraphic detection of cancer sites. Antibody 17-1A, or its F(ab')2 fragments, accumulated significantly in 27 of 46 (59%) colorectal cancer sites, but not in 21 nonepitheliomatous colon cancers and cancers at other sites. Antibody 19-9, or its F(ab')2 fragments, showed significant accumulation in 19 out of 29 (66%) colorectal cancer sites. In 17 patients, immunoscintigraphy with antibody 19-9 correlated with an immunoperoxidase study with the same antibody on resected tissue specimens. In 12 patients injected with two antibodies (17-1A + 19-9, or anti-CEA + 19-9), ten of 13 colorectal cancer sites were positive.


Thrombotic microangiopathy secondary to VEGF pathway inhibition by sunitinib.
Drugs targeting the VEGF pathway are associated with renal adverse events, including proteinuria, hypertension and thrombotic microangiopathy (TMA). Most cases of TMA are reported secondary to bevacizumab. It was shown recently that sunitinib, a small molecule inhibiting several tyrosine kinase receptors, including VEGF receptors, can also induce proteinuria, hypertension and biological features of TMA. Case. A 44-year-old woman with a history of malignant skin hidradenoma was started on sunitinib for refractory disease. She developed hypertension after 2 weeks and low-grade proteinuria after 4 weeks. Renal function remained normal, and biological signs of TMA were absent. A renal biopsy was performed 6 months later as proteinuria persisted, demonstrating typical features of TMA. The patient was given irbesartan, and sunitinib was continued for 3 months after diagnosis. Over this period, blood pressure and renal function remained stable and proteinuria became undetectable. We report on the first case of histologically documented TMA secondary to sunitinib and provide detailed description of renal histological involvement. This suggests that all anti-VEGF drugs may share a common risk for developing renal adverse events, including TMA. Our case highlights the possible discrepancy between mild clinical manifestation on one hand and severe TMA features on renal biopsy on the other hand and pleads for large indication of renal biopsy in this setting. The renin-angiotensin system blockers may be considered in patients with mild clinical manifestations and in the absence of therapeutic alternative to anti-VEGF drugs.


Prenatal rotation of the lumbar spine and its relevance for the development of the zygapophyseal joints.
The intrauterine axial rotational characteristics of the lumbar spine were analyzed and quantified sonographically. To determine whether axial rotational movements of the lumbar spine occur prenatally, which may influence the morphogenesis of the zygapophysial joints. Previously, the development of lumbar zygapophysial sagittal joint orientation was linked to the increase in torque of the torso as upright posture and locomotion are adopted. However, the potential influence of prenatal axial rotational movement of the lumbar spine on the development of the zygapophysial joints has not been examined. For this study, 52 healthy fetuses, gestation age 9 to 36 weeks, were monitored sonographically for rotational movements of the lumbar spine. By multiplanar reconstruction of the sonography images, exemplary segmental rotational values were acquired. This method was validated by directly measuring the manually induced rotation of the formalin-fixated fetus in addition to multiplanar reconstruction. Axial rotations of the lumbar spine were seen in most of the examined fetuses ages 9 to 36 gestation weeks. Rotations are best assessable with multiplanar reconstruction in the second trimenon, when segmental rotation of the lower lumbar spine reaches a mean of 8 degrees (range, 4-10 degrees ). Fetal axial rotation of the lower lumbar spine reaches a degree that may functionally influence the morphologic development of the articular processes of the zygapophysial joints. In contrast to the prevalent hypothesis, lumbar spine development is already functionally influenced prenatally by axial rotation.


Is there a role for FISH in the management and surveillance of patients with upper tract transitional-cell carcinoma?
Fluorescence in-situ hybridization (FISH) assay has been approved by the U.S. Food and Drug Administration for the detection of recurrent transitional-cell carcinoma (TCC) of the bladder and in the initial workup of hematuria. In this study, we retrospectively reviewed our initial 94 FISH specimens taken from patients monitored for upper-tract TCC. Between 2004 and 2007, 43 patients had one or more FISH assays performed as part of the workup and management of upper-tract TCC. Of 94 specimens sent for FISH analysis, 25 voided specimens collected at an outpatient encounter and 40 specimens taken as a bladder wash or selective upper-tract washing under anesthesia were followed by upper-tract endoscopy. The sensitivity and specificity of the FISH assay for detecting urothelial lesions in this population were calculated and compared with cytology specimens from the same sources. Overall sensitivity of FISH in the detection of TCC in this population was 52%, compared with 26% for urinary cytology. Both FISH and cytology showed superior sensitivity for high-grade (79% and 50%, respectively) nu low-grade tumors (41% and 12%, respectively). Selective upper-tract washings were more sensitive and specific for upper-tract TCC than bladder washings or voided specimens. While the sensitivity of FISH for upper-tract TCC parallels its performance in bladder cancer, the preponderance of low-grade, recurrent disease in the population undergoing surveillance and minimally invasive therapy for upper-tract TCC may limit its usefulness in this setting. Until a high-sensitivity marker for low-grade urothelial lesions is developed, the surveillance of upper-tract TCC will continue to require vigilant direct visual inspection.


Incidence of multidrug resistance and extended-spectrum beta-lactamase expression in community-acquired urinary tract infection among different age groups of patients.
Urinary tract infection (UTI) is a frequent disorder and depends on age and gender. Ineffective empiric treatment of UTI is common when associated with extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae. The aim of the study was to investigate the prevalence of Gram-negative uropathogens of E. coli and K. pneumoniae in different age groups along with the identification of ESBL-producing uropathogens and antimicrobial susceptibility profiles. A total of 247 uropathogens of E. coli and K. pneumoniae were collected over a period of 1 year (January-December 2015) from various diagnostic centers of Karachi city (Pakistan). Antimicrobial susceptibility analysis was performed by disc diffusion method, and identification of ESBL was performed by double disc synergy test. Categorical data of ESBL and non-ESBL uropathogens were analyzed by Pearson's Chi-square test. The study of 247 patients with community-acquired UTI comprised 72% females and 28% males, illustrating an increased prevalence of UTIs among females. It was also revealed that 90% belonged to the age group of 16-30 years whereas 78% related to the age group of 46-60 years. ESBL was found positive in 33.5% (63/188) of E. coli and 15.25% (9/59) in K. pneumonia, with a significant association i.e., (p =0.007). Amikacin, fosfomycin, imipenem, and tazobactam/piperacillin were found to be the effective treatment options. A significant association was found between ESBL-producing uropathogens against ciprofloxacin, enoxacin, and amoxicillin/clavulanic acid resistance (P < 0.05). It was concluded that for effective treatment of UTIs, appropriate screening of ESBL and culture sensitivity must be employed instead of empiric treatment.


Laser-assisted treatment of peri-implantitis: a retrospective cohort study.
The intent of this study was to provide a retrospective analysis of the clinical outcomes of 222 consecutive patients with 437 implants diagnosed with peri-implantitis and treated with the LAPIP protocol. All patients treated with LAPIP therapy at this practice were included. The primary outcome variable studied was probing depth (PD), and secondary variables were erythema, bleeding on probing, and suppuration. The significance of reductions in PD and clinical signs was assessed using repeated-measures analysis of variance. Complete data for both baseline and follow-up visits were available for 116 patients with a total of 224 treated implants. The rate of successful treatments-defined as follow-up PD ≤ 4.0 mm and elimination of clinical signs-was 90%. The reduction in PD from 5.4 mm at baseline to 3.4 mm at a median of 7.6 months was statistically significant (P ≤ 0.001). The reduction in the frequency of clinical signs was also statistically significant (P ≤ 0.001). Among 138 patients who had follow-up visits but not necessarily complete PD data, 15 implants were recorded as failed and 249 were recorded as intact at the median longest follow-up time of 13.1 months, resulting in a survival rate of 94%. In this single clinical practice, use of the minimally invasive LAPIP protocol for the treatment of peri-implantitis provided effective and predictable clinical outcomes. Future randomized controlled trials are indicated.


Dioptric defocus maps across the visual field for different indoor environments.
One of the factors proposed to regulate the eye growth is the error signal derived from the defocus in the retina and actually, this might arise from defocus not only in the fovea but the whole visual field. Therefore, myopia could be better predicted by spatio-temporally mapping the 'environmental defocus' over the visual field. At present, no devices are available that could provide this information. A 'Kinect sensor v1' camera (Microsoft Corp.) and a portable eye tracker were used for developing a system for quantifying 'indoor defocus error signals' across the central 58° of the visual field. Dioptric differences relative to the fovea (assumed to be in focus) were recorded over the visual field and 'defocus maps' were generated for various scenes and tasks.


Crouching variation revealed.
The term 'phenotypic capacitance' was introduced nearly 15 years ago to describe the strain-specific effects of impairing Hsp90, a molecular chaperone, in the fly Drosophila melanogaster (Rutherford & Lindquist 1998). In one genetic background, Hsp90 depletion caused deformed eyes, whereas in other genetic backgrounds, the wings or abdomens or other aspects of morphology were affected. Hsp90 was therefore viewed as acting like a capacitor, allowing genetic differences to build up and to be released at a later time. In the years since, it has been debated whether capacitance is a laboratory curiosity or a major force in evolution. In this issue of Molecular Ecology, Takahashi (2013) presents evidence, from high-resolution morphometric analysis of fly wings, that a large number of other capacitors exist in D. melanogaster, and that the variation they reveal can be quite subtle. His results advance our understanding of capacitance and contribute to a new view of its role in evolutionary adaptation.


Docosahexaenoic acid, protectins and dry eye.
To report recent data on the potential role of omega-3 fatty acids, in particular docosahexaenoic acid and its derivatives, in the treatment of dry eye syndrome. Dietary supplementation with polyunsaturated fatty acids yields positive results in the improvement of dry eye signs and symptoms. Although several studies have shown this, evidence is still lacking as to which fatty acid or what combination constitutes the most effective treatment. Studies show that treatment with alpha-linoleic acid reduces dry eye-induced inflammation. Eicosapentaenoic acid and docosahexaenoic acid derivatives, particularly resolvin E1 (RvE1) and neuroprotectin D1, appear to be responsible for docosahexaenoic acid's anti-inflammatory effect. This is supported in a study in which topical RvE1 resulted in decreased inflammation in a mouse dry eye model. Topical administration of pigment epithelium-derived factor in combination with docosahexaenoic acid accelerates the regeneration of corneal nerves after their damage during corneal surgery, promoting the return of sensitivity and reducing the signs of dry eye. This combined treatment also reduces objective signs of dry eye, such as rose bengal staining. No firm recommendations can be made regarding optimal dietary supplementation of essential fatty acids that benefit dry eye patients. On the basis of animal data and preliminary human studies, docosahexaenoic acid and its derivatives appear to be a safe, effective topical treatment for dry eye patients. This may result from their role in the resolution of inflammation and the regeneration of damaged corneal nerves.


Partial induction of the myogenic program in noncommitted adult stem cells.
The ability of multipotent adult stem cells (MASCs) and mesenchymal stem cells (MSCs) to acquire certain biochemical and functional properties of differentiated cells in response to various inductive cues has raised hopes to utilize such cells for therapeutic purposes. It is of particular importance to define precisely the differentiation status of induced MASCs and MSCs, and to determine the signaling pathways that control their differentiation. The identification of such signals might help to expand and enhance specific differentiation pathways to allow the formation of fully functional muscle cells from undetermined stem cells. Here, we have analyzed the ability of MASCs to contribute to the formation of muscle syncytia. We also investigated the intracellular signaling cascades that were initiated by Wnt molecules in a murine bone marrow-derived MASC population. We found that the partial recapitulation of the myogenic program depends on Wnt-mediated signaling via beta-catenin and protein kinase C, but not protein kinase A (PKA). Our experiments indicate that Wnt-mediated signaling via beta-catenin and protein kinase C (PKC) does not suffice to promote cell-autonomous differentiation of MASCs, but leads to acquisition of some properties of muscle cells that allow Wnt-induced cells to fuse to preexisting myotubes.


Important considerations for the clinical evaluation of drugs: appropriate dosing regimens.
A major aspect of clinical trials is the determination of a dosing regimen that will be effective and reasonably safe. The author presents a brief overview of the major considerations that govern the choice of dosage regimens for clinical trials and dosage recommendations for the labeling of marketed products. They include: (1) determination of the lowest effective dose, (2) determination of a safe dose for patients with liver or renal disease, (3) recommendations for doses in the elderly, (4) determination of the effect of concomitant drugs on drug disposition, and (5) examination of dose proportionality. Association of blood levels with desired therapeutic effect also is discussed.


Apolipoprotein D inhibits platelet-derived growth factor-BB-induced vascular smooth muscle cell proliferated by preventing translocation of phosphorylated extracellular signal regulated kinase 1/2 to the nucleus.
Elevated apolipoprotein D (apoD) levels are associated with reduced proliferation of cancer cells. We therefore investigated whether apoD, which occurs free or associated with HDL, suppresses vascular smooth muscle cell (VSMC) proliferation, which is related to the pathobiology of disease. Intense immunoreactivity for apoD was observed in human atherosclerotic plaque but not in normal coronary artery. However, an increase in apoD mRNA was seen in quiescent relative to proliferating fetal lamb aortic VSMCs, and in the rat aortic VSMC line (A10), we demonstrated uptake of apoD from serum. Stable transfection of apoD in A10 cells in the absence of serum did not influence VSMC proliferation assessed by [3H]-thymidine incorporation. ApoD, administered at a dose of 100 ng/mL, completely inhibited basal as well as platelet-derived growth factor (PDGF)-BB-induced VSMC proliferation (P<0.01) but had no effect on fibroblast growth factor-induced VSMC proliferation. ApoD did not suppress PDGF-BB or fibroblast growth factor-2-induced phosphorylation of extracellular signal regulated kinase (ERK) 1/2 but selectively inhibited PDGF-BB-mediated ERK1/2 nuclear translocation. Our data suggest that apoD selectively modulates the proliferative response of VSMC to growth factors by a mechanism related to nuclear translocation of ERK1/2.


Comparing Urinary Glycoproteins among Three Urogenital Cancers and Identifying Prostate Cancer-Specific Glycoproteins.
Prostate cancer, bladder cancer, and renal cancers are major urogenital cancers. Of which, prostate cancer is the most commonly diagnosed and second leading cause of cancer death for men in the United States. For urogenital cancers, urine is considered as proximate body fluid to the tumor site for developing non-invasiveness tests. However, the specific molecular signatures from different urogenital cancers are needed to relate changes in urine to various cancer detections. Herein, we utilized a previously published C4-Tip and C18/MAX-Tip workflow for enrichment of glycopeptides from urine samples and evaluated urinary glycopeptides for its cancer specificity. We analyzed 66 urine samples from bladder cancer (n = 27), prostate cancer (n = 4), clear cell renal cell carcinoma (ccRCC, n = 3), and benign plastic hyperplasia (BPH, n = 32) and then compared them with a previous publication that reported glycopeptides associated with aggressive prostate cancer (Gleason score ≥ 8). We further demonstrated the cancer specificity of the glycopeptides associated with aggressive prostate cancer. In this study, a total of 33 glycopeptides were identified to be specifically differentially expressed in prostate cancer compared to other urogenital cancer types as well as BPH urines. By cross-comparison with our previous urinary glycoproteomic dataset for aggressive prostate cancer, we reported a total of four glycopeptides from glycoproteins DSC2, MGAM, PIK3IP1, and CD55, commonly identified to be prostate cancer-specific. Together, these results deepen our understanding of the urinary glycoproteins associated with urogenital cancer types and expand our knowledge of the cancer specificity of urinary glycoproteins among urogenital cancer progression.


Novel magnetic-fluorescent bifunctional Janus nanofiber membrane.
Magnetic-fluorescent bifunctional materials have received global attention owing to their potential in many fields. Herein, we reported a novel magnetic-fluorescent bifunctional Janus nanofiber membrane (NFM) by adding the as-prepared magnetic CoFe2O4 nanoparticles into the polyacrylonitrile (PAN) side (m-PAN) and the fluorescent molecules of 1,8-naphthalene anhydride (1,8-NAD) into the polyvinylpyrrolidone (PVP) side (f-PVP) via electrospinning method. The obtained m-PAN/f-PVP Janus NFM exhibited excellent magnetic performance and high fluorescent properties due to the unique structure. Compared with the m-PAN/f-PVP composite NFM, the Janus NFM showed higher fluorescent performance because the fluorescent molecules were isolated from the magnetic nanoparticles. In addition, the Janus NFM not only maintain the good self-supporting state in water but also realize a directional movement attracted by a magnet. The unique structure of Janus nanofiber is of great importance and demonstrates great potential applications.


Optic disc and retinal nerve fiber layer parameters of eyes with keratoconus.
To evaluate optic nerve head (ONH) characteristics and retinal nerve fiber layer (RNFL) thickness measurements in patients with nonglaucomatous keratoconus. This was a cross-sectional, observational study in which 46 patients with keratoconus and 74 healthy subjects were included. Analysis of ONH and RNFL imaging were performed by confocal scanning laser ophthalmoscopy and optical coherence tomography, respectively. Eyes with keratoconus had larger disc area (2.37 ± 0.5 vs 2.17 ± 0.36 mm(2)) (P = .013), larger cup area (0.65 ± 0.53 vs 0.49 ± 0.28 mm(2)) (P = .035) and deeper cup depths (for mean cup depth 0.24 ± 0.09 vs 0.20 ± 0.07 mm, P = .008 and for maximum cup depth 0.69 ± 0.27 vs 0.60 ± 0.17 mm, P = .037) than control group eyes. Height variation contour also differed in the two groups (P = .007) but RNFL measurements did not. RNFL thickness between patients with keratoconus and healthy subjects seems to be more comparable than ONH parameters.


Novel anti-Sialyl-Tn monoclonal antibodies and antibody-drug conjugates demonstrate tumor specificity and anti-tumor activity.
Targeted therapeutics that can differentiate between normal and malignant tumor cells represent the ideal standard for the development of a successful anti-cancer strategy. The Sialyl-Thomsen-nouveau antigen (STn or Sialyl-Tn, also known as CD175s) is rarely seen in normal adult tissues, but it is abundantly expressed in many types of human epithelial cancers. We have identified novel antibodies that specifically target with high affinity the STn glycan independent of its carrier protein, affording the potential to recognize a wider array of cancer-specific sialylated proteins. A panel of murine monoclonal anti-STn therapeutic antibodies were generated and their binding specificity and efficacy were characterized in vitro and in in vivo murine cancer models. A subset of these antibodies were conjugated to monomethyl auristatin E (MMAE) to generate antibody-drug conjugates (ADCs). These ADCs demonstrated in vitro efficacy in STn-expressing cell lines and significant tumor growth inhibition in STn-expressing tumor xenograft cancer models with no evidence of overt toxicity.


Nocturnal antihypertensive treatment in patients with type 1 diabetes with autonomic neuropathy and non-dipping of blood pressure during night time: protocol for a randomised, placebo-controlled, double-blind, two-way crossover study.
Cardiac autonomic neuropathy (CAN) and elevated nocturnal blood pressure are independent risk factors for cardiovascular disease in patients with diabetes. Previously, associations between CAN, non-dipping of nocturnal blood pressure and coronary artery calcification have been demonstrated. The present protocol describes a trial to test the efficacy of bedtime dosing of the ACE inhibitor enalapril on night time blood pressure and left ventricular mass in patients with type 1 diabetes. In a randomised, double-blind, two-way cross-over study, 24 normoalbuminuric patients with type 1 diabetes with CAN will be treated for 12 weeks with either morning or bedtime dosing of 20 mg enalapril, followed by 12 weeks of switched treatment regimen. During each treatment period, two 24 h ambulatory blood pressure measurements will be performed and after each treatment period left ventricular mass will be determined by multisliced CT. Primary end points will be reduction in blood pressure and reduction in left ventricular mass. The study has been approved by the Danish Medicines Agency, the Scientific-Ethical Committee of the Capital Region of Denmark and the Danish Data Protection Agency. An external monitoring committee (the Good Clinical Practice Unit at Copenhagen University Hospital) will oversee the study. The results of the study will be presented at national and international scientific meetings and publications will be submitted to peer-reviewed journals. EudraCT (2012- 002136-90).


Thymus volume correlates with the progression of vertical HIV infection.
The thymus is the organ responsible for the maturation and selection of T lymphocytes and is thus pivotal in allowing the development of a functional immune system. Because in HIV infection cell-mediated immune responses are severely impaired, we studied the role of thymus in the control of the progression of HIV infection to AIDS. Thymic volume was analysed by magnetic resonance imaging in 31 vertically HIV-infected children. Plasma HIV viral load and phenotypic and functional cellular immunity-defining parameters were examined in the same patients. Thymic volume was not correlated with age or nutritional status; thymic volume was nevertheless correlated with CD4 T-lymphocyte counts and with the percentage and absolute number of CD45RA+CD62L+ (naive) T lymphocytes. In addition, the ability of peripheral blood mononuclear cells to proliferate upon tetanus stimulation was directly proportional to thymic volume. Finally, a negative correlation was detected between thymic volume and HIV viral load. Because low HIV plasma viraemia and preserved immune function are favourable prognostic indices in HIV disease, these data indicate that an immunological, thymic-dependent control of the progression of HIV infection might be possible, at least in vertically transmitted HIV infection.


Patterns of childhood abuse and neglect as predictors of treatment outcome in inpatient psychotherapy: a typological approach.
Childhood maltreatment is associated with the development and maintenance of mental disorders. The purpose of this naturalistic study was (a) to identify different patterns of childhood maltreatment, (b) to examine how these patterns are linked to the severity of mental disorders and (c) whether they are predictive of treatment outcome. 742 adult patients of a university hospital for psychotherapy and psychosomatics were assessed at intake and discharge by standardized questionnaires assessing depression (Beck Depression Inventory, BDI) and general mental distress (Symptom Check List-90-R, SCL-90-R). Traumatic childhood experience (using the Childhood Trauma Questionnaire, CTQ) and ICD-10 diagnoses were assessed at intake. The patients could be allocated to three different patterns of early childhood trauma experience: mild traumatization, multiple traumatization without sexual abuse and multiple traumatization with sexual abuse. The three patterns showed highly significant differences in BDI, General Severity Index (GSI) and in the number of comorbidity at intake. For both BDI and GSI a general decrease in depression and general mental distress from intake to discharge could be shown. The three patterns differed in BDI and GSI at intake and discharge, indicating lowest values for mild traumatization and highest values for multiple traumatization with sexual abuse. Patients with multiple traumatization with sexual abuse showed the least favourable outcome. The results provide evidence that the severity of childhood traumatization is linked to the severity of mental disorders and also to the treatment outcome in inpatient psychotherapy. In the study, three different patterns of childhood traumatization (mild traumatization, multiple traumatization without sexual abuse, multiple traumatization with sexual abuse) showed differences in the severity of mental disorder and in the course of treatment within the same therapy setting.


Regulation of ABCA1-mediated cholesterol efflux by sphingosine-1-phosphate signaling in macrophages.
Sphingolipid and cholesterol metabolism are closely associated at the structural, biochemical, and functional levels. Although HDL-associated sphingosine-1-phosphate (S1P) contributes to several HDL functions, and S1P signaling regulates glucose and lipid metabolism, no study has addressed the involvement of S1P in cholesterol efflux. Here, we show that sphingosine kinase (Sphk) activity was induced by the LXR agonist 22(R)-hydroxycholesterol and required for the stimulation of ABCA1-mediated cholesterol efflux to apolipoprotein A-I. In support, pharmacological Sphk inhibition and Sphk2 but not Sphk1 deficiency abrogated efflux. The involved mechanism included stimulation of both transcriptional and functional ABCA1 regulatory pathways and depended for the latter on the S1P receptor 3 (S1P3). Accordingly, S1P3-deficient macrophages were resistant to 22(R)-hydroxycholesterol-stimulated cholesterol efflux. The inability of excess exogenous S1P to further increase efflux was consistent with tonic S1P3 signaling by a pool of constitutively generated Sphk-derived S1P dynamically regulating cholesterol efflux. In summary, we have established S1P as a previously unrecognized intermediate in LXR-stimulated ABCA1-mediated cholesterol efflux and identified S1P/S1P3 signaling as a positive-feedback regulator of cholesterol efflux. This constitutes a novel regulatory mechanism of cholesterol efflux by sphingolipids.


The coefficient of dominance is not (always) estimable with biallelic markers.
The genetic relationship among individuals at one locus is characterized by nine coefficients of identity. The coefficients of inbreeding, coancestry and dominance (or fraternity) are just linear functions of them. Here, it is shown how they can be estimated using biallelic and triallelic markers using the method of moments, and comparisons are made with other methods based on molecular coancestry or molecular covariance. It is concluded that in the general case of dominance and inbreeding with biallelic markers, only the coefficients of inbreeding and coancestry can be estimated, but neither the single coefficients of identity nor the coefficient of dominance can be estimated. More than two alleles are required for a full estimation as illustrated with the triallelic situation.


Synthesis and structure of new compounds with Zn-Ga bonds: insertion of the gallium(I) bisimidinate Ga(DDP) into Zn-X (X = CH3, Cl) and the homoleptic complex cation [Zn(GaCp*)4]2+.
Insertion reactions of the low-valent group 13 bisimidinate ligand Ga(DDP) {DDP = 2-[(2,6-diisopropylphenyl)amino]-4-[(2,6-diisopropylphenyl)imino]-2-pentene} into Zn-Me and Zn-Cl bonds are reported. The reaction of ZnMe2 with 2 equiv of Ga(DDP) yields the double-insertion product [{(DDP)GaMe}2Zn] (1), whereas the insertion of Ga(DDP) into the Zn-Cl bond of ZnCl2 in tetrahydrofuran (THF) leads to the monoinsertion product [{(DDP)GaCl}ZnCl(THF)2] (2). Treatment of 2 with Na[BArF] results in the salt [{THF.Ga(DDP)}Zn(THF)(mu-Cl)]2[BArF]2 (3), with two Cl atoms bridging the Zn centers. The structural features of the Zn-Ga-bonded compounds 1-3 were compared with related complexes and in particular with the compound [Zn(GaCp*)4][BArF]2 (4), which was synthesized by the reaction of ZnMe2, [H(OEt2)2][BArF], and GaCp* in fluorobenzene. The complex cation [Zn(GaCp*)4]2+ of 4 relates to previously reported d10 analogues [M(GaCp*)4] (M = Ni, Pd, Pt). All new compounds were fully characterized by elemental analysis, NMR spectroscopy, and single-crystal X-ray diffraction analysis.


Garlic inhibits microsomal triglyceride transfer protein gene expression in human liver and intestinal cell lines and in rat intestine.
Epidemiologic studies have suggested that fresh garlic has lipid-lowering activity. Because the microsomal triglyceride transfer protein (MTP) plays a pivotal role in the assembly and secretion of apolipoprotein B (apoB)-containing lipoproteins, we evaluated the effect of garlic on the expression of the MTP gene in vitro in cell lines and in vivo in rats. Fresh garlic extract (FGE) reduced MTP mRNA levels in both the human hepatoma HepG2 and intestinal carcinoma Caco-2 cells in dose-dependent fashion; significant reductions were detected with 3 g/L FGE. Maximal 72 and 59% reductions, respectively, were observed with 6 g/L FGE. To evaluate the in vivo effect of garlic on MTP gene expression, rats were given a single oral dose of fresh garlic homogenate (FGH), with hepatic and intestinal MTP mRNA measured 3 h after dosing. Rats fed FGH had significantly (46% of the control) lower intestinal MTP mRNA levels compared with the control rats, whereas hepatic MTP mRNA levels were not affected. These results suggest a new mechanism for the hypolipidemic effect of fresh garlic. Long-term dietary supplementation of fresh garlic may exert a lipid-lowering effect partly through reducing intestinal MTP gene expression, thus suppressing the assembly and secretion of chylomicrons from intestine to the blood circulation.


Efficacy and Safety of Traditional Chinese Medicine in Coronavirus Disease 2019 (COVID-19): A Systematic Review and Meta-Analysis.
Introduction: Until now, there is no clinically approved specific medicine to treat COVID-19. Prior systematic reviews (SRs) have shown that traditional Chinese medicine (TCM) reduces the number of patients with severe disease and time to fever clearance, promotes clinical effectiveness, and improves chest images and the negativity rate of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleic acid test. Few SRs arrived at a definitive conclusion, and more randomized controlled trials (RCTs) were published. We conducted this study to summarize the latest evidence of TCM in COVID-19. Methods: Eight online databases were searched from December 2019 to July 2020, updated to March 2021. Only RCTs evaluating the clinical efficacy and safety of TCM in the treatment of COVID-19 were included. Primary outcomes were clinical cure and the negativity of the SARS-CoV-2 nucleic acid test. Secondary outcomes included clinical deterioration, ARDS, mechanical ventilation, death, time to fever clearance, duration of hospitalization, and chest imaging improvement. Safety outcomes included adverse events and serious adverse events during treatment. Two reviewers selected the included articles, assessed the risk of bias, and extracted data independently and in duplicate. Results: A total of 25 RCTs involving 2222 participants were selected in the systematic review, and seven RCTs were included in the meta-analysis. The results showed that TCM plus routine treatment was significantly better than routine treatment alone in clinical cure (risk ratio [RR] = 1.20, 95% confidence interval (CI) [1.04, 1.38], P = 0.01) and chest image improvement (RR = 1.22, 95% CI [1.07, 1.39], P = 0.01) and could reduce clinical deterioration (RR = 0.39, 95% CI [0.18, 0.86], P = 0.02), ARDS (RR = 0.28, 95% CI [0.11, 0.69], P = 0.01), mechanical ventilation (RR = 0.30, 95% CI [0.12, 0.77], P = 0.01), or death rate (RR = 0.28, 95% CI [0.09, 0.84], P = 0.02). No significant difference between TCM and routine treatment in the negativity of SARS-CoV-2 nucleic acid test (RR = 1.08, 95% CI [0.94, 1.23], P = 0.29) was observed. Finally, there was no overall significant difference in the incidence of adverse events between the two groups. The summary of evidence showed moderate confidence of a benefit of 11.8% in clinical cure and 14.0% in chest image improvement and a reduction of 5.9% in clinical deterioration, 25.4% in ARDS, 18.3% in mechanical ventilation, and 4.5% in death with TCM plus routine treatment compared to routine treatment alone in patients with COVID-19. A low confidence of a benefit of 5.4% in the negativity of SARS-CoV-2 nucleic acid test was also observed. Conclusions: Synethized evidence of 21 outcomes in 8 RCTs showed moderate certainty that TCM treatment plus routine treatment may promote a clinical cure and chest image improvement compared to routine treatment alone while reducing clinical deterioration, development of ARDS, use of mechanical ventilation, and death in patients with COVID-19. TCM treatment plus routine treatment may not promote the negativity of the SARS-CoV-2 nucleic acid test compared to routine treatment alone. TCM treatment was found to be safe for patients with COVID-19.


Opposite regulation of IL-1beta and secreted IL-1 receptor antagonist production by phosphatidylinositide-3 kinases in human monocytes activated by lipopolysaccharides or contact with T cells.
The unbalanced production of IL-1beta and its natural, specific inhibitor, the secreted IL-1R antagonist (sIL-1Ra), plays an important role in chronic/sterile inflammation. Relevant to this condition is direct cellular contact with stimulated T cells which is a potent inducer of cytokine production in human monocytes/macrophages. We previously demonstrated that activation of PI3Ks is a prerequisite of the transcription of the sIL-1Ra gene in human monocytes activated by IFN-beta. In this study, we addressed the question of PI3K involvement in the production of IL-1beta and sIL-1Ra in monocytes activated by cellular contact with stimulated T cells (mimicked by CHAPS-solubilized membranes of stimulated T cells (CE(sHUT))), and a crude preparation of LPS, to compare stimuli relevant to chronic/sterile and acute/infectious inflammation, respectively. In monocytes activated by either CE(sHUT) or LPS, the inhibition of PI3Ks abrogated sIL-1Ra transcript expression and sIL-1Ra production, demonstrating that PI3Ks control the induction of sIL-1Ra gene transcription. In contrast, PI3K inhibition increased the production of IL-1beta protein in both CE(sHUT)- and LPS-activated monocytes, the enhancement being drastically higher in the former. This was not due to changes in IL-1beta mRNA steady-state levels or transcript stability, but to the involvement of PI3Ks in the repression of IL-1beta secretion. The downstream PI3K effector, Akt, was implicated in this process. The present results demonstrate that PI3Ks are involved in the inhibition of IL-1beta secretion and in the induction of sIL-1Ra production in human blood monocytes by controlling different mechanisms in conditions mimicking chronic/sterile (CE(sHUT)) and acute/infectious (LPS) inflammation.


Angiotensin converting enzyme inhibitors and aortic arch obstructive malformations.
We describe two newborn infants with aortic arch obstructive malformations who became anuric after initiation of captopril. Since angiotensin converting enzyme inhibitors can alter renal blood flow by reduction in angiotensin II and blocking autoregulation phenomenon, it is important to use them with great caution in neonates with aortic arch obstructive malformations, while monitoring their renal function closely.


Novel Midkine Inhibitor iMDK Inhibits Tumor Growth and Angiogenesis in Oral Squamous Cell Carcinoma.
Midkine is a heparin-binding growth factor highly expressed in various human malignant tumors. However, its role in the growth of oral squamous cell carcinoma is not well understood. In this study, we analyzed the antitumor effect of a novel midkine inhibitor (iMDK) against oral squamous cell carcinoma. Administration of iMDK induced a robust antitumor response and suppressed cluster of differentiation 31 (CD31) expression in oral squamous cell carcinoma HSC-2 cells and SAS cells xenograft models. iMDK inhibited the proliferation of these cells dose-dependently, as well as the expression of midkine and phospho-extracellular signal-regulated kinase in HSC-2 and SAS cells. Moreover, iMDK significantly inhibited vascular endothelial growth factor and induced tube growth of human umbilical vein endothelial cells in a dose-dependent fashion. These findings suggest that midkine is critically involved in oral squamous cell carcinoma and iMDK can be effectively used for the treatment of oral squamous cell carcinoma.


Individual and flock immunity responses of naïve ducks on smallholder farms after vaccination with H5N1 Avian Influenza vaccine: a study in a province of the Mekong Delta, Vietnam.
In Vietnam, vaccination has played a crucial role in the national strategy for the prevention and control of H5 highly pathogenic avian influenza (HPAI). This study aimed to evaluate antibody responses of immunologically naïve domestic ducks to H5N1 avian influenza vaccine currently used in the national mass vaccination program of Vietnam. Blood samples of 166 ducks reared on smallholder farms were individually collected at three sampling time points, namely, right before vaccination, 21 days after primary vaccination, and 21 days after booster vaccination. Vaccine-induced antibody titers of duck sera were measured by the hemagglutination inhibition assay. Temporal differences in mean antibody titers were analyzed using the generalized least-squares method. No sampled ducks showed anti-H5 seropositivity pre-vaccination. The geometric mean titer (GMT) of the vaccinated ducks was 5.30 after primary vaccination, with 80% of the vaccinated ducks showing seropositivity. This result indicates that the immunity of duck flocks met the targets of the national poultry H5N1 HPAI mass vaccination program. GMT and seropositive rates of the ducks were 6.48 and 96.3%, respectively, after booster vaccination, which were significantly higher than those after primary vaccination. Flock-level seroprotection rate significantly increased from 68% to 84.7%, whereas variability in GMT titers decreased from 34.87% to 26.3%. This study provided important information on humoral immune responses of ducks to the currently used H5N1 vaccine under field conditions. Our findings may help guide veterinary authorities in planning effective vaccine protocols for the prevention and control of H5N1 in the target poultry population.


Relationship between Surface Hydroxyl Complexation and Equi-Acidity Point pH of MnO2 and Its Adsorption for Co2+ and Ni2.
MnO2 has shown great potential in the field of adsorption and has a good adsorption effect on heavy metal ions in aqueous solution, but there have been problems in the adsorption of heavy metal ions in high-concentration metal salt solutions. In this paper, different crystal forms of MnO2 (α-MnO2, β-MnO2, γ-MnO2, δ1-MnO2, δ2-MnO2, and ε-MnO2) were prepared and characterized by XRD, SEM, EDS, XPS, ZETA, and FT-IR. The reasons for the equi-acidity point pH change of MnO2 and the complex mechanism of surface hydroxylation on metal ions were discussed. The results showed that the equi-acidity point pHs of different crystalline MnO2 were different. The equi-acidity point pH decreased with the increase of reaction temperature and electrolyte concentration, but the reaction time had no effect on it. The equi-acidity point pHs of MnO2 were essentially equal to the equilibrium pH values of adsorption and desorption between surface hydroxyl and metal ions on them. The change of equi-acidity points was mainly due to the complexation of surface hydroxyl, and the equi-acidity point pHs depended on the content of surface hydroxyl and the size of the complexation ability. According to the equi-acidity point pH characteristics of MnO2, more hydroxyl groups could participate in the complexation reaction by repeatedly controlling the pH, so that MnO2 could adsorb heavy metals Co2+ and Ni2+ in high-concentration MnSO4 solution, and the adsorption rates of Co2+ and Ni2+ could reach 96.55 and 79.73%, respectively. The effects of MnO2 dosage and Mn2+ concentration on the adsorption performance were further investigated, and the products after MnO2 adsorption were analyzed by EDS and FT-IR. A new process for MnO2 to adsorb heavy metals Co2+ and Ni2+ in high-concentration MnSO4 solution was explored, which provided a reference for the deep purification of manganese sulfate solutions.


Ovarian granulosa cell-derived insulin-like growth factor binding proteins: modulatory role of follicle-stimulating hormone.
Recent observations disclosed the multiplicity of granulosa cell-derived high affinity insulin-like growth factor binding proteins (IGFBPs) and revealed the striking ability of high doses of FSH to suppress their constitutive release under both in vitro and in vivo circumstances. It is the objective of this communication to characterize in greater detail the modulatory effect(s) exerted by FSH on the elaboration of IGFBPs by granulosa cells from immature, estrogen-primed rats. The ability of FSH to regulate the elaboration of granulosa cell-derived IGFBPs proved dose-dependent but biphasic in nature. Specifically, FSH concentrations in the range of 1-3 ng/ml applied for 72 h produced a significant (P less than 0.05) increase in polyethylene glycol-precipitable [125I]IGF-I binding activity corresponding to all IGFBP species detectable by ligand blotting. In contrast, treatment with higher concentrations (greater than or equal to 10 ng/ml) of FSH resulted in progressive dose-dependent inhibition of the constitutive release of IGF-I binding activity (80% inhibition at the 100 ng/ml dose level) and the virtual elimination of all detectable IGFBP species. Time-course studies disclosed a significant (P less than 0.05) initial (apparent at the 24-h time point) stimulatory effect of a high dose of FSH (100 ng/ml) corresponding mostly (but not solely) to the single minor (23K) IGFBP band. In contrast, more prolonged exposure to FSH (greater than or equal to 48 h) produced progressive time-dependent decrements in IGF-I binding activity, an effect associated with a decrease in the relative representation of the major band doublet (28-29K), the 23K species being all but eliminated under these experimental circumstances. Hypophysectomy produced significant (P less than 0.05) inhibition of the subsequent in vitro release of granulosa cell-derived precipitable IGF-I binding activity strongly suggesting that (presumptively stimulatory) pituitary principles other than FSH are likely involved in the regulation of granulosa cell IGFBP release and that the trophic influence of these putative agents appears to outweight the potential disinhibition of FSH hormonal action. Taken together, these findings indicate that the pituitary regulation of granulosa cell-derived IGFBPs is complex and is not FSH-exclusive. Our findings further indicate that the ability of FSH to regulate granulosa cell-derived IGFBPs is dose- and time-dependent but biphasic in nature, an effect characterized by an early low-dose stimulatory effect and a late high-dose inhibitory effect.(ABSTRACT TRUNCATED AT 400 WORDS)


Progress toward the Total Synthesis of Lymphostins: Preparation of a Functionalized Tetrahydropyrrolo[4,3,2-de]quinoline and Unusual Oxidative Dimerization.
The lymphostins are a family of closely related pyrrolo[4,3,2-de]quinoline natural products produced by Streptomyces and Salinispora actinobacteria. Neolymphostin A was recently shown to strongly inhibit phosphoinositide 3-kinase (PI3K) and the mammalian target of rapamycin (mTOR) in a covalent manner via conjugation to a catalytic lysine residue in the ATP-binding pocket of the enzymes, making this metabolite the first reported covalent kinase inhibitor from a bacterium. A flexible and efficient synthetic route toward these alkaloids would allow for improvements in their solubility, stability, and selectivity and help to deliver a viable drug candidate. We have since established a short synthesis to methyl 8-bromo-1,3,4,5-tetrahydropyrrolo[4,3,2-de]quinoline-4-carboxylate via a conjugate addition/intramolecular Ullman reaction sequence. However, attempts to oxidize this intermediate to the pyrrolo[4,3,2-de]quinoline characteristic of the lymphostins resulted in formation of either a 2-oxo-1,2-dihydropyrrolo[4,3,2-de]quinoline or an unusual N,C-linked tetrahydropyrroloquinoline-pyrroloquinoline heterodimer. We expect that key modifications to the tetrahydropyrroloquinoline intermediate prior to oxidation should prevent these side reactions and pave the way for the completion of the synthesis.


Using Community Engagement to Develop a Web-Based Intervention for Latinos about the HPV Vaccine.
Human papillomavirus (HPV) infection is pervasive among sexually active women and men, and Hispanic women are at particularly high risk as they have higher rates of invasive cervical cancer compared to other racial or ethnic groups in the United States. There is a need for interventions to increase HPV vaccination among this high-risk population. This study investigated how to modify a previously developed web-based intervention that provided individually tailored information about HPV to improve its use among the Latino population. A community-oriented modification approach incorporated feedback from a community advisory committee, and focus groups among the Latino population, to modify the intervention. Several themes emerged including a need for basic information about HPV and HPV vaccination, changes to make the intervention appear less clinical, and incorporation of information addressing barriers specific to the Latino community. This work was done in preparation for a randomized trial to assess the impact of this modified intervention on HPV vaccination attitudes and uptake among Latino young adults and parents of adolescents. If effective, our intervention could be a resource for reducing HPV vaccination concerns, improving immunization rates, and educating Latinos about HPV and the HPV vaccine outside of the time boundaries of the traditional clinical encounter.


The combination of multi-approach studies to explore the potential therapeutic mechanisms of imidazole derivatives as an MCF-7 inhibitor in therapeutic strategies.
Breast cancer covers a large area of research because of its prevalence and high frequency all over the world. This study is based on drug discovery against breast cancer from a series of imidazole derivatives. A 3D-QSAR and activity atlas model was developed by exploring the dataset computationally, using the machine learning process of Flare. The dataset of compounds was divided into active and inactive compounds according to their biological and structural similarity with the reference drug. The obtained PLS regression model provided an acceptable r 2 = 0.81 and q2 = 0.51. Protein-ligand interactions of active molecules were shown by molecular docking against six potential targets, namely, TTK, HER2, GR, NUDT5, MTHFS, and NQO2. Then, toxicity risk parameters were evaluated for hit compounds. Finally, after all these screening processes, compound C10 was recognized as the best-hit compound. This study identified a new inhibitor C10 against cancer and provided evidence-based knowledge to discover more analogs.


In-die evaluation of capping tendency of pharmaceutical tablets using force-displacement curve and stress relaxation parameter.
A novel in-die evaluation method of tablet capping tendency was proposed based on a force-displacement curve and stress relaxation parameter in a tableting process. In our previous study (Chem. Pharm. Bull., 59, 2011, Nakamura et al.), the phase diagram consisting of elastic recovery energy (E(e)) and plastic deformation energy (E(p)) of compressed powder, named as the E(e)-E(p) diagram, was proposed. However, it was found that capping tendency of tablets prepared by double-compression with multi-component powder formulations cannot be discriminated using the E(e)-E(p) diagram. To improve the capping discrimination ability, we here proposed a novel corrected phase diagram consisting of the E(e) and an interparticle bonding parameter E(b)(t), named as the E(e)-E(b)(t) diagram. The E(b)(t) was proposed as a new parameter expressing strength of the interparticle bonding formed by the stress relaxation inside compressed powder. The E(b)(t) was defined as a product of the E(p) and the stress relaxation parameter Y(t), estimated from the force-displacement curve and the stress relaxation test. The capping discrimination ability of the diagrams was evaluated using a hierarchical-clustering analysis. The results exhibited that the capping tendency could be clearly discriminated using the proposed E(e)-E(b)(t) diagram at the double-compression and the multi-component powder formulations, as compared to the E(e)-E(p) diagram. This proposed diagram can be used for screening of the powder formulations to avoid the capping.


Histopathology and viral antigen distribution in lymph nodes of cats naturally infected with feline immunodeficiency virus.
Lymph nodes from cats with natural feline immunodeficiency virus (FIV) infection were examined histopathologically for characterization of lesions in the symptomatic and terminal stages. The localization of viral antigen was also studied in the nodes by immunohistochemistry using a monoclonal antibody to FIV p24. In a node of persistent generalized lymphadenopathy stage, follicular hyperplasia was prominent and the viral antigen was demonstrated predominantly in the follicular lymphocytes and tingible body macrophages. In cats died at AIDS related complex stage, the lymph nodes showed either mixed follicular hyperplasia and involution, involution, or mixed involution and depletion. The lymph nodes in those died at AIDS stage showed considerable destruction of the nodal architecture with involution and depletion of lymphoid follicles. In this terminal stage, the viral antigen was seen prominently in histiocytes/macrophages of the sinus. These findings observed in the dead cats were similar to the lymph node changes seen in human AIDS associated with HIV infection. It was suggested that sequential histologic changes from follicular hyperplasia to depletion via involution took place in the FIV infected lymph node with persistence of FIV antigen.


Economic burden and co-morbidities of atrial fibrillation in a privately insured population.
This study assesses the direct (medical and drugs) and indirect (work loss) annual costs associated with Atrial Fibrillation (AFIB) in a privately insured population. AFIB is the most common sustained rhythm disturbance, affecting 2.3 million people in the United States. Cost and co-morbidity measures for AFIB patients were compared to matched controls using a privately insured administrative database (including medical, drug, and disability claims) for 2 million enrollees, 1999-2002 from 16 employers across the United States. Patients with an AFIB diagnosis on at least two occasions were included in the co-morbidity analysis (n = 3944). A non-AFIB control sample was randomly selected with a 1:1 ratio, with characteristics (i.e., age, gender, health plan) matched to AFIB patients. Excess medical costs (i.e., employer payments) were estimated for AFIB patients, defined as the difference in average annual costs between AFIB and control patients (n = 3944); excess work-loss costs were defined similarly for employees with available work-loss data (n = 603). Statistical significance in the descriptive analysis was measured by paired t-tests for cost, or Chi-square tests for co-morbidity comparisons. A two-part multivariate model of excess cost was further estimated to control for co-existing conditions and other patient characteristics. The excess costs of AFIB patients were estimated as the difference between the observed costs of AFIB patients and their estimated costs, assuming they did not have AFIB. The multivariate analysis found that the excess annual direct cost of AFIB was $12,349 (p < 0.01), with AFIB patients approximately 5 times as costly as non-AFIB individuals ($15,553 versus $3204, respectively). The excess annual total cost was $14,875 (p < 0.01), with AFIB patients again approximately 5 times as costly as non-AFIB individuals ($18,454 versus $3,579, respectively). AFIB was associated with increased incidence of atrial flutter (p < 0.01), heart failure (Relative Risk (RR) = 29, p < 0.01), other arrhythmias/conduction disorders (RR = 16, p < 0.01), heart attack (RR = 8, p < 0.01), and stroke (RR = 6, p < 0.01). AFIB is a costly disease and one for which more definitive therapies might offer some potential for reducing, not only the clinical impact, but also the economic burden of the disease.


ANARCI: antigen receptor numbering and receptor classification.
Antibody amino-acid sequences can be numbered to identify equivalent positions. Such annotations are valuable for antibody sequence comparison, protein structure modelling and engineering. Multiple different numbering schemes exist, they vary in the nomenclature they use to annotate residue positions, their definitions of position equivalence and their popularity within different scientific disciplines. However, currently no publicly available software exists that can apply all the most widely used schemes or for which an executable can be obtained under an open license. ANARCI is a tool to classify and number antibody and T-cell receptor amino-acid variable domain sequences. It can annotate sequences with the five most popular numbering schemes: Kabat, Chothia, Enhanced Chothia, IMGT and AHo. ANARCI is available for download under GPLv3 license at opig.stats.ox.ac.uk/webapps/anarci. A web-interface to the program is available at the same address. email@example.com.


Induction of anthocyanin in the inner epidermis of red onion leaves by environmental stimuli and transient expression of transcription factors.
Novel imaging approaches have allowed measurements of the anthocyanin induction in onion epidermal cells that can be induced through water stress or transient expression of exogenous transcription factors. Environmental and genetic mechanisms that allow the normally colourless inner epidermal cells of red onion (Allium cepa) bulbs to accumulate anthocyanin were quantified by both absorbance ratios and fluorescence. We observed that water-stressing excised leaf segments induced anthocyanin formation, and fluorescence indicated that this anthocyanin was spectrally similar to the anthocyanin in the outer epidermal cells. This environmental induction may require a signal emanating from the leaf mesophyll, as induction did not occur in detached epidermal peels. Exogenous transcription factors that successfully drive anthocyanin biosynthesis in other species were also tested through transient gene expression using particle bombardment. Although the petunia R2R3-MYB factor AN2 induced anthocyanin in both excised leaves and epidermal peels, several transcription factors including maize C1 and Lc inhibited normal anthocyanin development in excised leaves. This inhibition may be due to moderate levels of conservation between the exogenous transcription factors and endogenous Allium transcription factors. The over-expressed exogenous transcription factors cannot drive anthocyanin biosynthesis themselves, but bind to the endogenous transcription factors and prevent them from driving anthocyanin biosynthesis.


Culture, Ethnicity, and Level of Education in Alzheimer's Disease.
Alzheimer's disease (AD) is the most frequent cause of dementia, where the abnormal accumulation of beta-amyloid (Aβ) and tau lead to neurodegeneration as well as loss of cognitive, behavioral, and functional abilities. The present review analyzes AD from a cross-cultural neuropsychological perspective, looking at differences in culture-associated variables, neuropsychological test performance and biomarkers across ethnic and racial groups. Studies have found significant effects of culture, preferred language, country of origin, race, and ethnicity on cognitive test performance, although the definition of those grouping terms varies across studies. Together, with the substantial underrepresentation of minority groups in research, the inconsistent classification might conduce to an inaccuratte diagnosis that often results from biases in testing procedures that favor the group to which test developers belong. These biases persist even after adjusting for variables related to disadvantageous societal conditions, such as low level of education, unfavorable socioeconomic status, health care access, or psychological stressors. All too frequently, educational level is confounded with culture. Minorities often have lower educational attainment and lower quality of education, causing differences in test results that are then attributed to culture. Higher levels of education are also associated with increased cognitive reserve, a protective factor against cognitive decline in the presence of neurodegeneration. Biomarker research suggests there might be significant differences in specific biomarker profiles for each ethnicity/race in need of accurate cultural definitions to adequately predict risk and disease progression across ethnic/racial groups. Overall, this review highlights the need for diversity in all domains of AD research that lack inclusion and the collection of relevant information from these groups.


How the O2-dependent Mg-protoporphyrin monomethyl ester cyclase forms the fifth ring of chlorophylls.
Mg-protoporphyrin IX monomethyl ester (MgPME) cyclase catalyses the formation of the isocyclic ring, producing protochlorophyllide a and contributing substantially to the absorption properties of chlorophylls and bacteriochlorophylls. The O2-dependent cyclase is found in both oxygenic phototrophs and some purple bacteria. We overproduced the simplest form of the cyclase, AcsF, from Rubrivivax gelatinosus, in Escherichia coli. In biochemical assays the di-iron cluster within AcsF is reduced by ferredoxin furnished by NADPH and ferredoxin:NADP+ reductase, or by direct coupling to Photosystem I photochemistry, linking cyclase to the photosynthetic electron transport chain. Kinetic analyses yielded a turnover number of 0.9 min-1, a Michaelis-Menten constant of 7.0 µM for MgPME and a dissociation constant for MgPME of 0.16 µM. Mass spectrometry identified 131-hydroxy-MgPME and 131-keto-MgPME as cyclase reaction intermediates, revealing the steps that form the isocyclic ring and completing the work originated by Sam Granick in 1950.


A Novel Mutation in SLC7A9 Gene in Cystinuria.
Cystinuria is an inherited disorder affecting luminal transport of cystine and dibasic amino acids. Because of the poor solubility of cystine in urine, stone formation in the kidney occurs frequently. Cystinuria is associated with mutations in the SLC3A1 and SLC7A9 genes. Despite the population-specific distribution of mutations in the SLC7A9 genes, there are few genetic data reported for cystinuric patients from the Middle East. Exon 4 of the SLC7A9 gene was sequenced in 21 patients with cystinuria, using the polymerase chain reaction and sequencing methods. A new variation in exon 4 of the SLC7A9 gene was identified, which was insertion of 1 adenine nucleotide between 2 cytosine nucleotides in position c.213-214insA. It seems to be important since it causes frame shift and it may be an important cause to make disease.


Hematologic and biochemical effects of the Gradiflow in an ex vivo ovine model.
The Gradiflow (Life Therapeutics, Frenchs Forest, Australia) system is a novel electrophoretic technique that uses the dual characteristics of size and charge to separate target macromolecules from complex biological solutions. The system has the potential to selectively remove a range of moieties from blood and plasma in an in vivo system such as hemodialysis or, alternatively, in an in vitro setting such as a blood bank. In this study, the safety of a scaled down Gradiflow prototype device with a membrane surface area of 16 cm2 was investigated using an ex vivo ovine model. Physiologic, hematologic, and biochemical parameters were assessed in 12 sheep: 6 animals treated using the Gradiflow and 6 controls. The effects of the Gradiflow procedure on both whole blood and plasma were analyzed. The Gradiflow procedure was well tolerated, and the application of an electrical potential or exposure to the Gradiflow membrane did not cause any significant changes in the parameters measured. Hemoglobin levels remained stable in all groups during the 4-hour experiment. An early neutropenia was observed in all groups, although this appeared to be more pronounced with exposure to a plasma filter; the presence of the Gradiflow component had no separate influence. One sheep in the plasma group experienced septic shock, caused by Staphylococcus contamination of the separation membrane. Overall, the results indicate that there were no gross physiologic, hematologic, or biochemical adverse reactions to the ex vivo Gradiflow procedure.