First Report of Bacterial Canker of Walnut Caused by Brenneria nigrifluens in France.
Since the summer of 2000, vertical oozing cankers have been observed on trunks and branches of Persian walnut trees (Juglans regia). Cvs. Fernor, Chandler, Mayette, and Hartley were the most frequently affected, but cvs. Lara and Franquette could also be affected. Brenneria nigrifluens (synonym Erwinia nigrifluens) (3) was isolated from diseased trees from 13 orchards and nurseries in southwestern (Aquitaine, Périgord, Charentes, and Quercy), southeastern (Grenoble areas), and western (near Angers) France. Cankers were observed on trunks and branches where brown-to-black exudates staining the bark appeared mainly in the summer. Isolations were performed primarily from exudates but also from infected tissues by using King's medium B. Colonies similar in appearance to Brenneria nigrifluens (1) were purified and characterized. Gram reaction, Kovac's oxidase, oxidative-fermentative metabolism, and urease activity were assayed for all isolates. API Biotype 100 kits (BioMérieux, Marcy l'Etoile, France) were used as recommended, except that incubations were at 28°C for 4 days. When compared with the reference strain (French Collection of Plant Pathogenic Bacteria (CFBP) 4998T = National Collection of Plant Pathogenic Bacteria (NCPPB) 564T = American Type Culture Collection (ATCC) 13028T) from California, 14 isolated strains were identified as B. nigrifluens on the basis of physiological and biochemical characteristics. These 14 strains were deposited in the CFBP under Accession Nos. 6746 to 6759. Pathogenicity of three selected strains (CFBP 6746, 6747, and 6758) was confirmed by inoculating branches of 7-year-old walnut trees with 108 CFU of each isolate introduced in wounds (2). The reference strain (CFBP 4998T) and water were similarly inoculated as controls. Two and five months later, necrotic lesions were observed in the inner bark and dark lines were observed in internal wood, but no external cankers were observed on any trees inoculated with the local and reference strains. B. nigrifluens was reisolated from the dark lines in internal wood up to approximately 10 cm from the inoculation site. To our knowledge, this is the first report of this bacterium in France. References: (1) L. Hauben et al. Syst. Appl. Microbiol. 21:384, 1998. (2) M. Ridé and S. Ridé. Proc. Int. Conf. Plant Pathogenic Bacteria, 4th, 2:957, 1978. (3) E. E. Wilson et al. Phytopathology 47:669, 1957.


Effect of gender on the association of adiposity with cardiovascular reactivity in Gujarati Indian adolescents.
Studies have reported that both gender and adiposity influence cardiovascular reactivity amongst adolescents. However, not much is known about the effect of gender on the association of adiposity with cardiovascular reactivity. The current study was conducted to learn how gender affects the association of adiposity with cardiovascular reactivity in the Gujarati Indian adolescents so as to develop preventive strategies for the local population. A cross-sectional study was conducted on 428 (Girls-173, Boys-255) Gujarati Indian adolescents of age group 16-19 years. Adiposity was assessed in terms of Body Mass Index (BMI), Body Fat Percentage (BF %), Fat Mass (FM), Fat Mass Index (FMI) and Waist Circumference (WC). Percentage Rise in Pulse Rate (%RPR) and Percentage Rise in Diastolic Blood Pressure (%RDBP) during Isometric Handgrip Test were used to assess the cardiovascular reactivity to acute sympathetic stress. Pearson's correlation coefficient was determined to find the association of adiposity with cardiovascular reactivity. Boys were found to have a significantly larger WC, higher physical fitness and greater %RDBP in comparison to girls while girls had a significantly higher BF% and FMI in comparison to boys. In both boys and girls, BMI, BF%, FMI and WC showed significant positive association with %RDBP with stronger correlationship found in girls. Girls also showed a significant negative correlationship between physical fitness and vascular reactivity. It could thus be concluded that gender affected the association of adiposity with vascular reactivity in Gujarati Indian adolescent such that girls tend to have a larger correlationship between adiposity and vascular reactivity than boys which is most likely due to lower physical fitness.


In vivo correlates of infectious salmon anemia virus pathogenesis in fish.
The phenotypic correlates of pathogenicity for Infectious salmon anemia virus (ISAV) in salmonid fishes have not been thoroughly studied to date. In this study, a comparison was made of 13 different strains of ISAV, isolated from different geographical regions between 1997 and 2004, for their infectivity in three fish species [Atlantic salmon (Salmo salar), coho salmon (Oncorhynchus kisutch) and rainbow trout (Oncorhynchus mykiss)]. When the different virus isolates were used at an approximate inoculum dose of 10(6) TCID(50) in 0.2 ml per fish, it was found that the most virulent strains had an acute mortality phase in Atlantic salmon that started at 10-13 days post-inoculation and lasted for 9-15 days with a cumulative mortality of >/=90 %. These highly pathogenic strains also caused low mortality in rainbow trout, albeit later in infection. Viruses with a more delayed or protracted mortality phase resulting in cumulative mortalities of 50-89 % in Atlantic salmon were considered to be of intermediate pathogenicity and isolates with cumulative mortalities of </=49 % were considered to be of low pathogenicity. On this basis, three of the ISAV isolates showed a high-, eight an intermediate- and two a low-pathogenicity phenotype in Atlantic salmon. Coho salmon were resistant to all ISAV isolates. These results confirmed that there is variation in pathogenicity among ISAV strains for Atlantic salmon and rainbow trout, and that other salmonid species such as coho salmon can carry highly pathogenic strains of ISAV without showing signs of disease. The identified pathogenicity phenotypes may aid in the identification of molecular markers of ISAV virulence.


Speckle reduction in synthetic-aperture-radar imagery.
Speckle appearing in synthetic-aperture-radar images degrades the information contained in these images. Speckle noise can be suppressed by adapted local processing techniques, permitting the definition of statistical parameters inside a small window centered on each pixel of the image. Two processing algorithms are examined; the first one uses the intensity as a variable, and the second one works on a homomorphic transformation of the image intensity. A statistical model for speckle noise that takes into account correlation in multilook imagery has been used to develop these processing algorithms. Several experimental results of processed Seasat-A syntheticaperture-radar images are discussed.


Effects of Zn(2+), Ca(2+), and Mg(2+) on the structure of Zn(7)metallothionein-3: evidence for an additional zinc binding site.
Human metallothionein-3 (Zn(7)MT-3), an intra- and extracellularly occurring metalloprotein, is highly expressed in the brain, where it plays an important role in the homeostasis of the essential metal ions Cu(+) and Zn(2+). Like other mammalian metallothioneins (MT-1 and -2), the protein contains a M(II)(3)(CysS)(9) and a M(II)(4)(CysS)(11) cluster localized in two independent protein domains linked by a flexible hinge region. However, there is a substantially increased number of acidic residues in MT-3 (11 residues) compared with MT-2 (four residues) which may act as binding ligands for additional metal ions. In this study, the binding of Zn(2+), Ca(2+), and Mg(2+) to human Zn(7)MT-3 and its mutant lacking an acidic hexapeptide insert, Zn(7)MT-3(Delta55-60), was investigated and compared with the binding of Zn(7)MT-2. By using spectroscopic and spectrometric techniques, we demonstrate that one additional Zn(2+) binds with an apparent binding constant (K(app)) of approximately 100 microM to Zn(7)MT-3 and Zn(7)MT-3(Delta55-60), but not to Zn(7)MT-2. The changes in spectroscopic features of metal-thiolate clusters and gel filtration behavior reveal that the formation of Zn(8)MT-3 is immediate and is accompanied by a decrease in the Stokes radius (R(s)). The changes in the R(s) suggest a mutual approach of both protein domains. The fast binding of Zn(2+) is followed by a slow time-dependent protein dimerization. The binding of Zn(2+) to Zn(7)MT-3 is specific as in the presence of Ca(2+) and Mg(2+) only an alteration of the R(s) of Zn(7)MT-3 at substantially higher concentrations was observed. The significance of these findings for the biological role of MT-3 is discussed.


Real-world evidence studies into treatment adherence, thresholds for intervention and disparities in treatment in people with type 2 diabetes in the UK.
The University of Surrey-Lilly Real World Evidence (RWE) diabetes cohort has been established to provide insights into the management of type 2 diabetes mellitus (T2DM). There are 3 areas of study due to be conducted to provide insights into T2DM management: exploration of medication adherence, thresholds for changing diabetes therapies, and ethnicity-related or socioeconomic-related disparities in management. This paper describes the identification of a cohort of people with T2DM which will be used for these analyses, through a case finding algorithm, and describes the characteristics of the identified cohort. A cohort of people with T2DM was identified from the Royal College of General Practitioners Research and Surveillance Centre (RCGP RSC) data set. This data set comprises electronic patient records collected from a nationally distributed sample of 130 primary care practices across England with scope to increase the number of practices to 200. A cohort (N=58 717) of adults with T2DM was identified from the RCGP RSC population (N=1 260 761), a crude prevalence of diabetes of 5.8% in the adult population. High data quality within the practice network and an ontological approach to classification resulted in a high level of data completeness in the T2DM cohort; ethnicity identification (82.1%), smoking status (99.3%), alcohol use (93.3%), glycated haemoglobin (HbA1c; 97.9%), body mass index (98.0%), blood pressure (99.4%), cholesterol (87.4%) and renal function (97.8%). Data completeness compares favourably to other, similarly large, observational cohorts. The cohort comprises a distribution of ages, socioeconomic and ethnic backgrounds, diabetes complications, and comorbidities, enabling the planned analyses. Regular data uploads from the RCGP RSC practice network will enable this cohort to be followed prospectively. We will investigate medication adherence, explore thresholds and triggers for changing diabetes therapies, and investigate any ethnicity-related or socioeconomic-related disparities in diabetes management.


Antimetastatic activities of heparins and modified heparins. Experimental evidence.
Heparin is commonly used for prevention or treatment of cancer-associated thromboembolism. Recent clinical evidence indicates that heparin, and low-molecular weight heparin improves survival of cancer patients. Experimental evidence from various animal models consistently supports the ability of heparin to attenuate metastasis. Heparin, apart from its anticoagulant activity contains a variety of biological activities possibly affecting cancer progression, including: inhibition of heparanase, blocking of P- and L-selectin mediated cell adhesion, and inhibition of angiogenesis. The delineation of antimetastatic activity of heparin is in the focus of several ongoing investigations. This review summarizes the current experimental evidence on the biology of heparin as a potential treatment cancer progression.


To B or Not to B a flower: the role of DEFICIENS and GLOBOSA orthologs in the evolution of the angiosperms.
DEFICIENS (DEF) and GLOBOSA (GLO) function in petal and stamen organ identity in Antirrhinum and are orthologs of APETALA3 and PISTILLATA in Arabidopsis. These genes are known as B-function genes for their role in the ABC genetic model of floral organ identity. Phylogenetic analyses show that DEF and GLO are closely related paralogs, having originated from a gene duplication event after the separation of the lineages leading to the extant gymnosperms and the extant angiosperms. Several additional gene duplications followed, providing multiple potential opportunities for functional divergence. In most angiosperms studied to date, genes in the DEF/GLO MADS-box subfamily are expressed in the petals and stamens during flower development. However, in some angiosperms, the expression of DEF and GLO orthologs are occasionally observed in the first and fourth whorls of flowers or in nonfloral organs, where their function is unknown. In this article we review what is known about function, phylogeny, and expression in the DEF/GLO subfamily to examine their evolution in the angiosperms. Our analyses demonstrate that although the primary role of the DEF/GLO subfamily appears to be in specifying the stamens and inner perianth, several examples of potential sub- and neofunctionalization are observed.


Abnormal bone microarchitecture and evidence of osteoblast dysfunction in premenopausal women with idiopathic osteoporosis.
Idiopathic osteoporosis (IOP) in premenopausal women is an uncommon disorder of uncertain pathogenesis in which fragility fractures occur in otherwise healthy women with intact gonadal function. It is unclear whether women with idiopathic low bone mineral density and no history of fragility fractures have osteoporosis. The objective of the study was to elucidate the microarchitectural and remodeling features of premenopausal women with IOP. We performed transiliac biopsies after tetracycline labeling in 104 women: 45 with fragility fractures (IOP), 19 with idiopathic low bone mineral density (Z score ≤-2.0) and 40 controls. Biopsies were analyzed by two-dimensional quantitative histomorphometry and three-dimensional microcomputed tomography. Bone stiffness was estimated using finite element analysis. Compared with controls, affected women had thinner cortices; fewer, thinner, more widely separated, and heterogeneously distributed trabeculae; reduced stiffness; and lower osteoid width and mean wall width. All parameters were indistinguishable between women with IOP and idiopathic low bone mineral density. Although there were no group differences in dynamic histomorphometric remodeling parameters, serum calciotropic hormones, bone turnover markers, or IGF-I, subjects in the lowest tertile of bone formation rate had significantly lower osteoid and wall width, more severely disrupted microarchitecture, lower stiffness, and higher serum IGF-I than those in the upper two tertiles, suggesting that women with low turnover IOP have osteoblast dysfunction with resistance to IGF-I. Subjects with high bone turnover had significantly higher serum 1,25 dihydroxyvitamin D levels and a nonsignificant trend toward higher serum PTH and urinary calcium excretion. These results suggest that the diagnosis of IOP should not require a history of fracture. Women with IOP may have high, normal or low bone turnover; those with low bone turnover have the most marked deficits in microarchitecture and stiffness. These results also suggest that the pathogenesis of idiopathic osteoporosis is heterogeneous and may differ according to remodeling activity.


Phosphorylation of Leukotriene C4 Synthase at Serine 36 Impairs Catalytic Activity.
Leukotriene C4 synthase (LTC4S) catalyzes the formation of the proinflammatory lipid mediator leukotriene C4 (LTC4). LTC4 is the parent molecule of the cysteinyl leukotrienes, which are recognized for their pathogenic role in asthma and allergic diseases. Cellular LTC4S activity is suppressed by PKC-mediated phosphorylation, and recently a downstream p70S6k was shown to play an important role in this process. Here, we identified Ser(36) as the major p70S6k phosphorylation site, along with a low frequency site at Thr(40), using an in vitro phosphorylation assay combined with mass spectrometry. The functional consequences of p70S6k phosphorylation were tested with the phosphomimetic mutant S36E, which displayed only about 20% (20 μmol/min/mg) of the activity of WT enzyme (95 μmol/min/mg), whereas the enzyme activity of T40E was not significantly affected. The enzyme activity of S36E increased linearly with increasing LTA4 concentrations during the steady-state kinetics analysis, indicating poor lipid substrate binding. The Ser(36) is located in a loop region close to the entrance of the proposed substrate binding pocket. Comparative molecular dynamics indicated that Ser(36) upon phosphorylation will pull the first luminal loop of LTC4S toward the neighboring subunit of the functional homotrimer, thereby forming hydrogen bonds with Arg(104) in the adjacent subunit. Because Arg(104) is a key catalytic residue responsible for stabilization of the glutathione thiolate anion, this phosphorylation-induced interaction leads to a reduction of the catalytic activity. In addition, the positional shift of the loop and its interaction with the neighboring subunit affect active site access. Thus, our mutational and kinetic data, together with molecular simulations, suggest that phosphorylation of Ser(36) inhibits the catalytic function of LTC4S by interference with the catalytic machinery.


Potentially preventable hospitalizations - United States, 2001-2009.
Potentially preventable hospitalizations are admissions to a hospital for certain acute illnesses (e.g., dehydration) or worsening chronic conditions (e.g., diabetes) that might not have required hospitalization had these conditions been managed successfully by primary care providers in outpatient settings. Although not all such hospitalizations can be avoided, admission rates in populations and communities can vary depending on access to primary care, care-seeking behaviors, and the quality of care available. Because hospitalization tends to be costlier than outpatient or primary care, potentially preventable hospitalizations often are tracked as markers of health system efficiency. The number and cost of potentially preventable hospitalizations also can be calculated to help identify potential cost savings associated with reducing these hospitalizations overall and for specific populations.


Monitoring Ceratomyxa shasta infection during a hatchery rearing cycle: comparison of molecular, serological and histological methods.
The prevalence of Ceratomyxa shasta infection in production stocks of steelhead Oncorhynchus mykiss and cutthroat trout O. clarki was monitored using a parasite-specific polymerase chain reaction (PCR) assay. For all 4 stocks of fish followed through their 1 yr rearing cycle, C. shasta infection was detected despite their genetic resistance to the disease and the treatment of the incoming water with ozone. Infection was confirmed using serological methods and standard histological procedures, except when prevalence was low (<10%). This suggests that at the lowest infection levels PCR is more sensitive than other methodologies, and can be used as an early indicator of infection. Results of the PCR assay continued to correlate with histological and serological detection as the numbers of parasites and the lesion severity increased over the rearing cycle. For both steelhead and cutthroat trout, early infections were characterized by large numbers of parasites on the epithelial surface, but with little associated inflammation. At release as yearlings, the infection prevalence in all stocks was greater than 90 % and the inflammatory response in many fish was extensive, with tissue necrosis and mucosal damage. Although C. shasta infections no longer result in high mortality at this facility, results of this study indicate that the parasite remains a contributor to low condition indices in these fish, despite their genetic resistance and ozone disinfection of the water supply.


Genomic mechanisms of climate adaptation in polyploid bioenergy switchgrass.
Long-term climate change and periodic environmental extremes threaten food and fuel security1 and global crop productivity2-4. Although molecular and adaptive breeding strategies can buffer the effects of climatic stress and improve crop resilience5, these approaches require sufficient knowledge of the genes that underlie productivity and adaptation6-knowledge that has been limited to a small number of well-studied model systems. Here we present the assembly and annotation of the large and complex genome of the polyploid bioenergy crop switchgrass (Panicum virgatum). Analysis of biomass and survival among 732 resequenced genotypes, which were grown across 10 common gardens that span 1,800 km of latitude, jointly revealed extensive genomic evidence of climate adaptation. Climate-gene-biomass associations were abundant but varied considerably among deeply diverged gene pools. Furthermore, we found that gene flow accelerated climate adaptation during the postglacial colonization of northern habitats through introgression of alleles from a pre-adapted northern gene pool. The polyploid nature of switchgrass also enhanced adaptive potential through the fractionation of gene function, as there was an increased level of heritable genetic diversity on the nondominant subgenome. In addition to investigating patterns of climate adaptation, the genome resources and gene-trait associations developed here provide breeders with the necessary tools to increase switchgrass yield for the sustainable production of bioenergy.


Long-term results of hypofractionated radiation therapy for breast cancer.
The optimal fractionation schedule for whole-breast irradiation after breast-conserving surgery is unknown. We conducted a study to determine whether a hypofractionated 3-week schedule of whole-breast irradiation is as effective as a 5-week schedule. Women with invasive breast cancer who had undergone breast-conserving surgery and in whom resection margins were clear and axillary lymph nodes were negative were randomly assigned to receive whole-breast irradiation either at a standard dose of 50.0 Gy in 25 fractions over a period of 35 days (the control group) or at a dose of 42.5 Gy in 16 fractions over a period of 22 days (the hypofractionated-radiation group). The risk of local recurrence at 10 years was 6.7% among the 612 women assigned to standard irradiation as compared with 6.2% among the 622 women assigned to the hypofractionated regimen (absolute difference, 0.5 percentage points; 95% confidence interval [CI], -2.5 to 3.5). At 10 years, 71.3% of women in the control group as compared with 69.8% of the women in the hypofractionated-radiation group had a good or excellent cosmetic outcome (absolute difference, 1.5 percentage points; 95% CI, -6.9 to 9.8). Ten years after treatment, accelerated, hypofractionated whole-breast irradiation was not inferior to standard radiation treatment in women who had undergone breast-conserving surgery for invasive breast cancer with clear surgical margins and negative axillary nodes. (ClinicalTrials.gov number, NCT00156052.)


Loss of col8a1a function during zebrafish embryogenesis results in congenital vertebral malformations.
Congenital vertebral malformations (CVM) occur in 1 in 1000 live births and in many cases can cause spinal deformities, such as scoliosis, and result in disability and distress of affected individuals. Many severe forms of the disease, such as spondylocostal dystostosis, are recessive monogenic traits affecting somitogenesis, however the etiologies of the majority of CVM cases remain undetermined. Here we demonstrate that morphological defects of the notochord in zebrafish can generate congenital-type spine defects. We characterize three recessive zebrafish leviathan/col8a1a mutant alleles ((m531, vu41, vu105)) that disrupt collagen type VIII alpha1a (col8a1a), and cause folding of the embryonic notochord and consequently adult vertebral column malformations. Furthermore, we provide evidence that a transient loss of col8a1a function or inhibition of Lysyl oxidases with drugs during embryogenesis was sufficient to generate vertebral fusions and scoliosis in the adult spine. Using periodic imaging of individual zebrafish, we correlate focal notochord defects of the embryo with vertebral malformations (VM) in the adult. Finally, we show that bends and kinks in the notochord can lead to aberrant apposition of osteoblasts normally confined to well-segmented areas of the developing vertebral bodies. Our results afford a novel mechanism for the formation of VM, independent of defects of somitogenesis, resulting from aberrant bone deposition at regions of misshapen notochord tissue.


Transcatheter Aortic Valve Implantation for Degenerated Surgical Aortic Bioprosthesis: A Systematic Review.
Transcatheter aortic valve in valve (Aviv) replacement has been shown to be an effective therapeutic option in patients with failed aortic bioprosthetic valves. This review intended to evaluate contemporary 1-year outcomes of Aviv in recent studies. A systematic review on outcomes of Aviv was performed using the best available evidence from studies obtained using a MEDLINE, Cochrane database, and SCOPUS search. Endpoints of interest were survival, coronary artery obstruction, prosthesis-patient mismatch (PPM), stroke, pacemaker implantation, and structural valve deterioration. A total of 3339 patients from 23 studies were included. Mean age was 68-80 years, 20%-50% were female, and Society of Thoracic Surgeons score ranged from 5.7 to 31.1. Thirty-day all-cause mortality ranged from 2% to 8%, and 1-year all-cause mortality ranged from 8% to 33%. Coronary artery obstruction risk after Aviv ranged from 0.6% to 4%. One-year stroke ranged from 2% to 8%. Moderate-severe PPM occurred in 11%-58%, and pacemaker rate at 1 year ranged from 5% to 12%. Transcatheter aortic ViV has emerged as an effective therapeutic option to treat patients with failed bioprostheses. The acceptable complication rate and favorable 1-year outcomes make Aviv an appropriate alternative to redo surgical aortic valve replacement.


Too much time? Time use and fertility-specific quality of life among men and women seeking specialty care for infertility.
There are known gender differences in the impacts infertility has on quality of life and well-being. Less is known about how infertile couples spend time on fertility-related tasks and associations with quality of life. The purpose of this study is to evaluate whether time spent on tasks related to family-building decision-making (including research, reflection, discussions with partner, discussions with others, and logistics) were associated with fertility-specific quality of life or anxiety among new patients. Couples or individuals (N = 156) with upcoming initial consultations with a reproductive specialist completed the Fertility Quality of Life (FertiQoL) tool, which produces a Core (total) score and four subscales: Emotional, Relational, Social, and Mind-Body. We developed questions to measure time spent in the previous 24 h on tasks related to family-building. We tested for differences by gender in time use (McNemar's Test) and used ordinary least squares regression to analyze the relationship between time use and FertiQoL scores. In the week before a new consultation, a higher percentage of women reported time spent in the past 24 h in research, reflecting, discussion with others, and logistics compared to male partners (all p < 0.05). In adjusted models, more time spent reflecting was associated with worse FertiQoL scores for both men and women, as well as with higher anxiety for men. Time spent in discussion with others was associated with higher anxiety for women but better Social FertiQoL scores for men. Couples seeking infertility consultation with a specialist reported spending time on tasks related to family-building before the initial visit. There were gender differences in the amount of time spent on these tasks, and time was associated with fertility-specific quality of life and anxiety.


Acute right-sided heart failure caused by Neisseria meningitidis.
Meningococcal myocarditis is a rarely diagnosed infection and could be the consequence of primary invasive infection or late immunologic complications. An unusual presentation of meningococcemia in an immunocompetent adult is described, with Neisseria meningitidis identified as the cause of selective right-sided heart failure in a case of acute myocarditis.


Curcumin inhibition of integrin (alpha6beta4)-dependent breast cancer cell motility and invasion.
Curcumin, a polyphenol natural product isolated from the rhizome of the plant Curcuma longa, has emerged as a promising anticancer therapeutic agent. However, the mechanism by which curcumin inhibits cancer cell functions such as cell growth, survival, and cell motility is largely unknown. We explored whether curcumin affects the function of integrin alpha(6)beta(4), a laminin adhesion receptor with an established role in invasion and migration of cancer cells. Here we show that curcumin significantly reduced alpha(6)beta(4)-dependent breast cancer cell motility and invasion in a concentration-dependent manner without affecting apoptosis in MDA-MB-435/beta4 (beta(4)-integrin transfectants) and MDA-MB-231 breast cancer cell lines. Further, curcumin selectively reduced the basal phosphorylation of beta(4) integrin (Y1494), which has been reported to be essential in mediating alpha(6)beta(4)-dependent phosphatidylinositol 3-kinase activation and cell motility. Consistent with this finding, curcumin also blocked alpha(6)beta(4)-dependent Akt activation and expression of the cell motility-promoting factor ENPP2 in MDA-MB-435/beta4 cell line. A multimodality approach using curcumin in combination with other pharmacologic inhibitors of alpha(6)beta(4) signaling pathways showed an additive effect to block breast cancer cell motility and invasion. Taken together, these findings show that curcumin inhibits breast cancer cell motility and invasion by directly inhibiting the function of alpha(6)beta(4) integrin, and suggest that curcumin can serve as an effective therapeutic agent in tumors that overexpress alpha(6)beta(4).


Orbicularis oculi muscle biopsies for mitochondrial DNA analysis in suspected mitochondrial myopathy.
We wished to demonstrate the feasibility of performing diagnostic mitochondrial DNA (mtDNA) analysis on biopsies of the orbicularis oculi muscle in patients with a chronic progressive external ophthalmoplegia (CPEO) phenotype and suspicion of an underlying mitochondrial disorder. Case series of three patients who underwent ptosis surgery and had simultaneous biopsy of the orbicularis oculi muscle because of a suspicion of a mitochondrial disorder. Orbicularis muscle samples were divided into two pieces at the time of biopsy. The first was snap-frozen in liquid nitrogen, DNA was extracted and mtDNA deletion analysis was performed by two complementary methods (long PCR and Southern blot analysis). The second piece of muscle was assessed using routine histopathology, electron microscopy and immuno-histochemical analysis. Three patients with clinical features of CPEO, without any positive family history, underwent orbicularis muscle biopsies at time of eyelid ptosis surgery. All biopsies were adequate to conduct histopathological and immuno-histochemical analysis, which showed evidence of abnormal muscle structure and function. mtDNA was successfully extracted from all biopsies, and long PCR and Southern blot analysis confirmed diagnostic large single mtDNA deletions in all three cases. Orbicularis oculi muscle biopsies are useful in patients with CPEO to perform mtDNA analysis, thus avoiding a separate biopsy of skeletal muscle elsewhere.


Comparison of mechanical properties of various suture repair techniques in a partially lacerated tendon.
The technique of surgical repair for zone two flexor tendon injuries has been debated extensively throughout the years, yet adhesion formation, suture rupture, and suture locking on the pulley edge remain possible consequences of a poor repair. The partially lacerated tendon is especially challenging to treat since there can be justification for not intervening surgically. In a partial laceration canine model we measured failure load and suture gap formation for tendons repaired with the Lee, modified four-strand Savage, Kessler, modified Kessler, and Augmented Becker core suture techniques and with a simple running peripheral suture. The modified Kessler (106.3 N, SD 18.8 N) and modified Savage (108.2 N, SD 19.9 N) repair techniques had a significantly higher failure load than the Lee (85.0 N, SD 20.6 N) suture method (P < 0.05), while there were no differences among the other techniques. There were no significant differences in resistance to gap formation among the repair techniques, with the mean values ranging from 38.9 N/mm (SD 15.7 N/mm) using the simple running suture to 53.2 N/mm (SD 25.8 N/mm) with the Kessler repair. The mean load to produce a 1.5 mm repair site gap ranged from 71.1 N (SD 21.5 N) in the Lee repair to 91.3 N (SD 22.2 N) in the Augmented Becker repair although there were no significant differences among repair methods. All repair methods were much weaker than tendons left unrepaired (184.7 N, SD 41.3 N).


Exome genotyping arrays to identify rare and low frequency variants associated with epithelial ovarian cancer risk.
Rare and low frequency variants are not well covered in most germline genotyping arrays and are understudied in relation to epithelial ovarian cancer (EOC) risk. To address this gap, we used genotyping arrays targeting rarer protein-coding variation in 8,165 EOC cases and 11,619 controls from the international Ovarian Cancer Association Consortium (OCAC). Pooled association analyses were conducted at the variant and gene level for 98,543 variants directly genotyped through two exome genotyping projects. Only common variants that represent or are in strong linkage disequilibrium (LD) with previously-identified signals at established loci reached traditional thresholds for exome-wide significance (P < 5.0 × 10 - 7). One of the most significant signals (Pall histologies = 1.01 × 10 - 13;Pserous = 3.54 × 10 - 14) occurred at 3q25.31 for rs62273959, a missense variant mapping to the LEKR1 gene that is in LD (r2 = 0.90) with a previously identified 'best hit' (rs7651446) mapping to an intron of TIPARP. Suggestive associations (5.0 × 10 - 5 > P≥5.0 ×10 - 7) were detected for rare and low-frequency variants at 16 novel loci. Four rare missense variants were identified (ACTBL2 rs73757391 (5q11.2), BTD rs200337373 (3p25.1), KRT13 rs150321809 (17q21.2) and MC2R rs104894658 (18p11.21)), but only MC2R rs104894668 had a large effect size (OR = 9.66). Genes most strongly associated with EOC risk included ACTBL2 (PAML = 3.23 × 10 - 5; PSKAT-o = 9.23 × 10 - 4) and KRT13 (PAML = 1.67 × 10 - 4; PSKAT-o = 1.07 × 10 - 5), reaffirming variant-level analysis. In summary, this large study identified several rare and low-frequency variants and genes that may contribute to EOC susceptibility, albeit with possible small effects. Future studies that integrate epidemiology, sequencing, and functional assays are needed to further unravel the unexplained heritability and biology of this disease.


Candidate gene polymorphisms in patients with acetaminophen-induced acute liver failure.
Acetaminophen is a leading cause of acute liver failure (ALF). Genetic differences might predispose some individuals to develop ALF. In this exploratory study, we evaluated genotype frequency differences among patients enrolled by the ALF Study Group who had developed ALF either intentionally from a single-time-point overdose of acetaminophen (n = 78), unintentionally after chronic high doses of acetaminophen (n = 79), or from causes other than acetaminophen (n = 103). The polymorphisms evaluated included those in genes encoding putative acetaminophen-metabolizing enzymes (UGT1A1, UGT1A6, UGT1A9, UGT2B15, SULT1A1, CYP2E1, and CYP3A5) as well as CD44 and BHMT1. Individuals carrying the CYP3A5 rs776746 A allele were overrepresented among ALF patients who had intentionally overdosed with acetaminophen, with an odds ratio of 2.3 (95% confidence interval, 1.1-4.9; P = 0.034) compared with all other ALF patients. This finding is consistent with the enhanced bioactivation of acetaminophen by the CYP3A5 enzyme. Persons homozygous for the CD44 rs1467558 A allele were also overrepresented among patients who had unintentionally developed ALF from chronic acetaminophen use, with an odds ratio of 4.0 (1.0-17.2, P = 0.045) compared with all other ALF subjects. This finding confirms a prior study that found elevated serum liver enzyme levels in healthy volunteers with the CD44 rs1467558 AA genotype who had consumed high doses of acetaminophen for up to 2 weeks. However, both genetic associations were considered relatively weak, and they were not statistically significant after adjustment for multiple comparisons testing. Nevertheless, both CYP3A5 rs776746 and CD44 rs1467558 warrant further investigation as potential genomic markers of enhanced risk of acetaminophen-induced ALF.


GAT: a graph-theoretical analysis toolbox for analyzing between-group differences in large-scale structural and functional brain networks.
In recent years, graph theoretical analyses of neuroimaging data have increased our understanding of the organization of large-scale structural and functional brain networks. However, tools for pipeline application of graph theory for analyzing topology of brain networks is still lacking. In this report, we describe the development of a graph-analysis toolbox (GAT) that facilitates analysis and comparison of structural and functional network brain networks. GAT provides a graphical user interface (GUI) that facilitates construction and analysis of brain networks, comparison of regional and global topological properties between networks, analysis of network hub and modules, and analysis of resilience of the networks to random failure and targeted attacks. Area under a curve (AUC) and functional data analyses (FDA), in conjunction with permutation testing, is employed for testing the differences in network topologies; analyses that are less sensitive to the thresholding process. We demonstrated the capabilities of GAT by investigating the differences in the organization of regional gray-matter correlation networks in survivors of acute lymphoblastic leukemia (ALL) and healthy matched Controls (CON). The results revealed an alteration in small-world characteristics of the brain networks in the ALL survivors; an observation that confirm our hypothesis suggesting widespread neurobiological injury in ALL survivors. Along with demonstration of the capabilities of the GAT, this is the first report of altered large-scale structural brain networks in ALL survivors.


RGN-259 (thymosin β4) improves clinically important dry eye efficacies in comparison with prescription drugs in a dry eye model.
This study evaluated the clinical activity of RGN-259 (thymosin β4) in comparison with cyclosporine A (CsA), diquafosol (DQS), and lifitegrast (LFA) in a murine model of dry eye. The model was NOD.B10-H2b mice in a 30-40% humidified environment together with daily scopolamine hydrobromide injections for 10 days. After desiccation stress, all drugs were evaluated after 10 treatment days. RGN-259 increased tear production similar to that in the DQS- and LFA-treated mice while CsA was inactive. RGN-259 improved corneal smoothness and decreased fluorescein staining similar to that of LFA group while CsA and DQS were inactive. Corneal epithelial detachment was reduced by RGN-259, and DQS and LFA showed similar activity but the CsA was inactive. RGN-259 increased conjunctival goblet cells and mucin production comparable to that seen with CsA, while DQS and LFA were inactive. RGN-259 reduced the over-expression of inflammatory factors comparable to that with CsA and LFA, while DQS was inactive. RGN-259 increased mucin production comparable to that observed with CsA, while DQS and LFA were inactive. In conclusion, RGN-259 promoted recovery of mucins and goblet cells, improved corneal integrity, and reduced inflammation in a dry eye mouse model and was equal to or more effective than prescription treatments.


What memory is for.
Let's start from scratch in thinking about what memory is for, and consequently, how it works. Suppose that memory and conceptualization work in the service of perception and action. In this case, conceptualization is the encoding of patterns of possible physical interaction with a three-dimensional world. These patterns are constrained by the structure of the environment, the structure of our bodies, and memory. Thus, how we perceive and conceive of the environment is determined by the types of bodies we have. Such a memory would not have associations. Instead, how concepts become related (and what it means to be related) is determined by how separate patterns of actions can be combined given the constraints of our bodies. I call this combination "mesh." To avoid hallucination, conceptualization would normally be driven by the environment, and patterns of action from memory would play a supporting, but automatic, role. A significant human skill is learning to suppress the overriding contribution of the environment to conceptualization, thereby allowing memory to guide conceptualization. The effort used in suppressing input from the environment pays off by allowing prediction, recollective memory, and language comprehension. I review theoretical work in cognitive science and empirical work in memory and language comprehension that suggest that it may be possible to investigate connections between topics as disparate as infantile amnesia and mental-model theory.


Serological and molecular epidemiological study on swine influenza in Zambia.
Influenza A viruses (IAVs) cause highly contagious respiratory diseases in humans and animals. In 2009, a swine-origin pandemic H1N1 IAV, designated A(H1N1)pdm09 virus, spread worldwide, and has since frequently been introduced into pig populations. Since novel reassortant IAVs with pandemic potential may emerge in pigs, surveillance for IAV in pigs is therefore necessary not only for the pig industry but also for public health. However, epidemiological information on IAV infection of pigs in Africa remains sparse. In this study, we collected 246 serum and 605 nasal swab samples from pigs in Zambia during the years 2011-2018. Serological analyses revealed that 49% and 32% of the sera collected in 2011 were positive for hemagglutination-inhibition (HI) and neutralizing antibodies against A(H1N1)pdm09 virus, respectively, whereas less than 5.3% of sera collected during the following period (2012-2018) were positive in both serological tests. The positive rate and the neutralization titres to A(H1N1)pdm09 virus were higher than those to classical swine H1N1 and H1N2 IAVs. On the other hand, the positive rate for swine H3N2 IAV was very low in the pig population in Zambia in 2011-2018 (5.3% and 0% in HI and neutralization tests, respectively). From nasal swab samples, we isolated one H3N2 and eight H1N1 IAV strains with an isolation rate of 1.5%. Phylogenetic analyses of all eight gene segments revealed that the isolated IAVs were closely related to human IAV strains belonging to A(H1N1)pdm09 and seasonal H3N2 lineages. Our findings indicate that reverse zoonotic transmission from humans to pigs occurred during the study period in Zambia and highlight the need for continued surveillance to monitor the status of IAVs circulating in swine populations in Africa.


Prediction of spatial variation in global fallout of 137Cs using precipitation.
Deposition from atmospheric nuclear weapons tests (termed global fallout) has been shown to be proportional to the rate of precipitation. Here we describe methods for using precipitation and radionuclide deposition information for a reference site to estimate global fallout at other locations. These methods have been used to estimate global fallout in Iceland, identified during the Arctic Monitoring and Assessment Programme (AMAP) by Wright et al. [Wright, S.M., Howard, B.J., Strand, P., Nylén, T., Sickel, M.A.K., 1999. Prediction of 137Cs deposition from atmospheric nuclear weapons tests within the Arctic. Environ Pollut 104, 131-143.] as one of the Arctic areas which received the highest global fallout, but where measurements of contamination were sparse, and difficult to obtain due to the remote and inaccessible terrain of much of the country. Measurements of global fallout 137Cs deposition have been made in Iceland at sites close to meteorological stations to ensure that precipitation data were of high quality. The AMAP modeling approach, based on measured precipitation and radionuclide deposition data, was applied using a reference monitoring station located close to Reykjavik. The availability of good precipitation data and locally based estimates of time dependent ratios of 137Cs deposition to precipitation during the fallout period gave a better correlation between predicted and measured 137Cs global fallout (r2=0.96) than that achieved using the much more heterogeneous set of data collected by AMAP over the whole of the Arctic. Having obtained satisfactory results with the model for a number of calibration sites alongside meteorological stations we then produced a map of estimated 137Cs deposition based on a model of estimated precipitation. This deposition map was then successfully validated (r2=0.85) for sites where 137Cs deposition was measured; the associated uncertainty in predictions was also estimated.


Molecular diversity after a range expansion in heterogeneous environments.
Recent range expansions have probably occurred in many species, as they often happen after speciation events, after ice ages, or after the introduction of invasive species. While it has been shown that range expansions lead to patterns of molecular diversity distinct from those of a pure demographic expansion, the fact that many species do live in heterogeneous environments has not been taken into account. We develop here a model of range expansion with a spatial heterogeneity of the environment, which is modeled as a gamma distribution of the carrying capacities of the demes. By allowing temporal variation of these carrying capacities, our model becomes a new metapopulation model linking ecological parameters to molecular diversity. We show by extensive simulations that environmental heterogeneity induces a loss of genetic diversity within demes and increases the degree of population differentiation. We find that metapopulations with low average densities are much more affected by environmental heterogeneity than metapopulations with high average densities, which are relatively insensitive to spatial and temporal variations of the environment. Spatial heterogeneity is shown to have a larger impact on genetic diversity than temporal heterogeneity. Overall, temporal heterogeneity and local extinctions are not found to leave any specific signature on molecular diversity that cannot be produced by spatial heterogeneity.


What is the point of tips?
To look at the incidence of tip margin involvement in skin excisions for basal cell carcinomas (BCCs) and to examine if any factors might be used to predict margin involvement. All reports of BCCs by a single dermatopathologist were reviewed and 793 excisions were included from 642 patients. Whether there was BCC in the tip and whether this tip was involved were recorded together with macroscopic and microscopic factors based on the Royal College of Pathologists' (RCPath) dataset. Statistical analysis was carried out to determine associations between these factors and tip involvement. In 43 (5%) specimens the tip margin was involved and in 35 (4%) cases the peripheral lateral margin was involved. Risk factors for tip margin involvement were a lesion that was non-discernible macroscopically and most importantly, involvement of a peripheral lateral margin. The incidence of tip margin involvement is small and it appears unnecessary to embed the tips unless the lesion is non-discernible macroscopically or if the peripheral lateral margin is involved.


Long-term in vitro culture and characterisation of avian embryonic stem cells with multiple morphogenetic potentialities.
Petitte, J.N., Clarck, M.E., Verrinder Gibbins, A. M. and R. J. Etches (1990; Development 108, 185-189) demonstrated that chicken early blastoderm contains cells able to contribute to both somatic and germinal tissue when injected into a recipient embryo. However, these cells were neither identified nor maintained in vitro. Here, we show that chicken early blastoderm contains cells characterised as putative avian embryonic stem (ES) cells that can be maintained in vitro for long-term culture. These cells exhibit features similar to those of murine ES cells such as typical morphology, strong reactivity toward specific antibodies, cytokine-dependent extended proliferation and high telomerase activity. These cells also present high capacities to differentiate in vitro into various cell types including cells from ectodermic, mesodermic and endodermic lineages. Production of chimeras after injection of the cultivated cells reinforced the view that our culture system maintains in vitro some avian putative ES cells.


The BDGP gene disruption project: single transposon insertions associated with 40% of Drosophila genes.
The Berkeley Drosophila Genome Project (BDGP) strives to disrupt each Drosophila gene by the insertion of a single transposable element. As part of this effort, transposons in >30,000 fly strains were localized and analyzed relative to predicted Drosophila gene structures. Approximately 6300 lines that maximize genomic coverage were selected to be sent to the Bloomington Stock Center for public distribution, bringing the size of the BDGP gene disruption collection to 7140 lines. It now includes individual lines predicted to disrupt 5362 of the 13,666 currently annotated Drosophila genes (39%). Other lines contain an insertion at least 2 kb from others in the collection and likely mutate additional incompletely annotated or uncharacterized genes and chromosomal regulatory elements. The remaining strains contain insertions likely to disrupt alternative gene promoters or to allow gene misexpression. The expanded BDGP gene disruption collection provides a public resource that will facilitate the application of Drosophila genetics to diverse biological problems. Finally, the project reveals new insight into how transposons interact with a eukaryotic genome and helps define optimal strategies for using insertional mutagenesis as a genomic tool.


A phylum-level phylogenetic classification of zygomycete fungi based on genome-scale data.
Zygomycete fungi were classified as a single phylum, Zygomycota, based on sexual reproduction by zygospores, frequent asexual reproduction by sporangia, absence of multicellular sporocarps, and production of coenocytic hyphae, all with some exceptions. Molecular phylogenies based on one or a few genes did not support the monophyly of the phylum, however, and the phylum was subsequently abandoned. Here we present phylogenetic analyses of a genome-scale data set for 46 taxa, including 25 zygomycetes and 192 proteins, and we demonstrate that zygomycetes comprise two major clades that form a paraphyletic grade. A formal phylogenetic classification is proposed herein and includes two phyla, six subphyla, four classes and 16 orders. On the basis of these results, the phyla Mucoromycota and Zoopagomycota are circumscribed. Zoopagomycota comprises Entomophtoromycotina, Kickxellomycotina and Zoopagomycotina; it constitutes the earliest diverging lineage of zygomycetes and contains species that are primarily parasites and pathogens of small animals (e.g. amoeba, insects, etc.) and other fungi, i.e. mycoparasites. Mucoromycota comprises Glomeromycotina, Mortierellomycotina, and Mucoromycotina and is sister to Dikarya. It is the more derived clade of zygomycetes and mainly consists of mycorrhizal fungi, root endophytes, and decomposers of plant material. Evolution of trophic modes, morphology, and analysis of genome-scale data are discussed.


Fluid removal in acute heart failure: diuretics versus devices.
Fluid removal and relief of congestion are central to treatment of acute heart failure. Diuretics have been the decongestive mainstay but their known limitations have led to the exploration of alternative strategies. This review compares diuretics with ultrafiltration and examines the recent evidence evaluating their use. Relevant recent studies are the Diuretic Optimization Strategies Evaluation trial (of diuretics) and the Cardiorenal Rescue Study in Acute Decompensated Heart Failure (of ultrafiltration). The Diuretic Optimization Strategies Evaluation study evaluated strategies of loop diuretic use during acute heart failure (continuous infusion versus intermittent bolus and high dose versus low dose). After 72 h, there was no significant difference with either comparison for the coprimary end points. Patients treated with a high-dose strategy tended to have greater diuresis and more decongestion compared with low-dose therapy, at the cost of transient changes in renal function. The Cardiorenal Rescue Study in Acute Decompensated Heart Failure study showed that in acute heart failure patients with persistent congestion and worsening renal function, ultrafiltration, as compared with a medical therapy, was associated with similar weight loss but greater increase in serum creatinine and more adverse events. Decongestion remains a major challenge in acute heart failure. Although recent studies provide useful data to guide practice, the relatively poor outcomes point to the continued need to identify better strategies for safe and effective decongestion.


The Relative Importance of Family History, Gender, Mode of Onset, and Age at Onsetin Predicting Clinical Features of First-Episode Psychotic Disorders.
Family history of psychosis, gender, mode of onset, and age at onset are considered prognostic factors important to clinicians evaluating first-episode psychosis; yet, clinicians have little guidance as to how these four factors differentially predict early-course substance abuse, symptomatology, and functioning. We conducted a "head-to-head comparison" of these four factors regarding their associations with key clinical features at initial hospitalization. We also assessed potential interactions between gender and family history with regard to age at onset of psychosis and symptom severity. Consecutively admitted first-episode patients (n=334) were evaluated in two studies that rigorously assessed a number of early-course variables. Associations among variables of interest were examined using Pearson correlations, χ2 tests, Student's t-tests, and 2×2 factorial analyses of variance. Substance (nicotine, alcohol, and cannabis) abuse and positive symptom severity were predicted only by male gender. Negative symptom severity and global functioning impairments were predicted by earlier age at onset of psychosis. General psychopathology symptom severity was predicted by both mode of onset and age at onset. Interaction effects were not observed with regard to gender and family history in predicting age at onset or symptom severity. The four prognostic features have differential associations with substance abuse, domains of symptom severity, and global functioning. Gender and age at onset of psychosis appear to be more predictive of clinical features at the time of initial evaluation (and thus presumably longer term outcomes) than the presence of a family history of psychosis and a more gradual mode of onset.


Rheometer enabled study of cartilage frequency-dependent properties.
Despite the well-established dependence of cartilage mechanical properties on the frequency of the applied load, most research in the field is carried out in either load-free or constant load conditions because of the complexity of the equipment required for the determination of time-dependent properties. These simpler analyses provide a limited representation of cartilage properties thus greatly reducing the impact of the information gathered hindering the understanding of the mechanisms involved in this tissue replacement, development and pathology. More complex techniques could represent better investigative methods, but their uptake in cartilage research is limited by the highly specialised training required and cost of the equipment. There is, therefore, a clear need for alternative experimental approaches to cartilage testing to be deployed in research and clinical settings using more user-friendly and financial accessible devices. Frequency dependent material properties can be determined through rheometry that is an easy to use requiring a relatively inexpensive device; we present how a commercial rheometer can be adapted to determine the viscoelastic properties of articular cartilage. Frequency-sweep tests were run at various applied normal loads on immature, mature and trypsinased (as model of osteoarthritis) cartilage samples to determine the dynamic shear moduli (G*, G' G″) of the tissues. Moduli increased with increasing frequency and applied load; mature cartilage had generally the highest moduli and GAG depleted samples the lowest. Hydraulic permeability (KH) was estimated from the rheological data and decreased with applied load; GAG depleted cartilage exhibited higher hydraulic permeability than either immature or mature tissues. The rheometer-based methodology developed was validated by the close comparison of the rheometer-obtained cartilage characteristics (G*, G', G″, KH) with results obtained with more complex testing techniques available in literature. Rheometry is relatively simpler and does not require highly capital intensive machinery and staff training is more accessible; thus the use of a rheometer would represent a cost-effective approach for the determination of frequency-dependent properties of cartilage for more comprehensive and impactful results for both healthcare professional and R&D.


Distribution of HCN1 and HCN2 in rat auditory brainstem nuclei.
Auditory brainstem neurons that are involved in the precise analysis of the temporal pattern of sounds have ionic currents activated near the resting potential to shorten membrane time constants. One of these currents is the hyperpolarization-activated current (Ih). Molecular cloning of the channels underlying Ih revealed four different isoforms (HCN1-4). HCN1 and HCN2, which are widely distributed in the brain, differ in their activation kinetics, voltage dependence and sensitivity to cAMP. We determined the distribution of the HCN1 and HCN2 isoform in the auditory brainstem and midbrain of young rats (P20-30), using standard immunohistochemical techniques. HCN1 antibodies gave rise to punctate staining on the somatic and dendritic membrane. Strong HCN1 staining was present on octopus and bushy cells of the ventral cochlear nucleus, principal neurons of the lateral and medial superior olive, and neurons of the ventral nucleus of the lateral lemniscus. No HCN1 staining was observed in the dorsal cochlear nucleus and the medial nucleus of the trapezoid body (MNTB). In contrast, HCN2 staining was strongest in the MNTB and the dorsal nucleus of the lateral lemniscus. Strong HCN2 antibody labelling was also observed in bushy cells of the ventral cochlear nucleus. In the central nucleus of the inferior colliculus only a subpopulation of neurons showed HCN1 or HCN2 immunolabelling. This differential distribution of HCN1 and HCN2 channels is in agreement with the physiologically observed Ih currents in corresponding neuronal populations and might represent the basis for functional heterogeneity and diverse sensitivity to neuromodulators.


Structure and RNA recognition by the snRNA and snoRNA transport factor PHAX.
Small nuclear and small nucleolar RNAs (snRNAs and snoRNAs) are critical components of snRNPs and snoRNPs and play an essential role in the maturation of, respectively, mRNAs and rRNAs within the nucleus of eukaryotic cells. Complex and specific pathways exist for the assembly of snRNPs and snoRNPs, involving, for instance, nucleocytoplasmic transport of snRNAs and intranuclear transport between compartments of snoRNAs. The phosphorylated adaptor for nuclear export (PHAX) is required for nuclear export of snRNAs in metazoans and also involved in the intranuclear transport of snoRNAs to Cajal bodies. PHAX contains a conserved single-stranded nucleic acid binding domain (RNA_GG_bind domain) with no sequence homology with any other known RNA-binding module. Here, we report NMR and X-ray crystallography studies that elucidate the structural basis for RNA recognition by the PHAX RNA-binding domain (PHAX-RBD). The crystal structure of the RNA_GG_bind domain from the parasite Cryptosporidium parvum (Cp RBD) forms well-folded dimers in solution in the absence of any ligand. The human PHAX-RBD is monomeric and only adopts a tertiary fold upon RNA binding. The PHAX-RBD represents a novel helical fold and binds single-stranded RNA with micromolar affinity without sequence specificity. RNA recognition by human PHAX-RBD is consistent with mutational analysis that affects RNA binding and PHAX-mediated nuclear export. Our data suggest that the PHAX-RBD mediates auxiliary RNA contacts with the snRNA and snoRNA substrates that are required for transport and/or substrate release.


Molecular epidemiology of candidemia: evidence of clusters of smoldering nosocomial infections.
Invasive fungal infections pose a serious threat to hospitalized patients worldwide. In particular, the prevalence of clusters of nosocomial infection among patients with candidemia remains unknown. The aim of this study was to investigate the molecular epidemiology of candidemia in a nationwide setting in Iceland during a 16-year period. The genotypes of all available fungal bloodstream isolates during 1991-2006 (n = 219) were determined by polymerase chain reaction fingerprinting with use of 4 separate primers. Clusters were defined as isolation of > or =2 strains with genotypes that had > or =90% relatedness in the same hospital within a period of 90 days. Candida albicans represented 61.6% of isolates, followed by Candida glabrata (13.7%), Candida tropicalis (9.1%), and Candida parapsilosis (8.7%). Polymerase chain reaction fingerprinting revealed 35 clones of C. albicans, 10 clones of C. glabrata, 7 clones of C. tropicalis, 4 clones of C. parapsilosis, and 5 clones of Candida dubliniensis. Overall, 18.7%-39.9% of all infections were part of nosocomial clusters, most commonly caused by C. albicans, C. parapsilosis, and C. tropicalis. Most clusters involved 2 cases and disproportionately affected patients in adult and neonatal intensive care units (P = .045). The 7-day (16%) and 30-day (32%) case-fatality rates among cluster-associated cases did not differ statistically significantly from those for sporadic nosocomial infections. None of the clusters were identified by the hospital surveillance team. In an unselected patient population, as many as one-third of all cases of candidemia may be attributable to nosocomial clusters. The risk is dependent on hospital wards and patient populations; it is highest in intensive care units. Small clusters are not identified by routine hospital surveillance.


Mechanism of the pulmonary vasoconstrictor action of digoxin in the dog.
The pulmonary vascular effects of a subarrhythmic dose of digoxin (60 micrograms/kg i.v.) were examined in the canine in situ perfused lung. Digoxin produced an increase in pulmonary vascular resistance (66.1%) and pulmonary arterial pressure (8.2 mm Hg) at 70 min after injection in the constant-flow, blood-perfused lung preparation. The digoxin-treated group exhibited higher plasma levels of norepinephrine compared with control dogs. The pulmonary vasoconstrictor response to digoxin was abolished by prior treatment with the alpha-adrenergic antagonists phenoxybenzamine and phentolamine. This vasoconstriction does not involve inhibition of synthesis or action of vasodilator prostaglandins by digoxin, as pretreatment with indomethacin did not attenuate, and even tended to increase, the pressor response to digoxin. The response was prevented by prior treatment with blockers of nonneuronal uptake of catecholamines normetanephrine and hydrocortisone, but not with cocaine, a blocker of neuronal uptake. In the lung preparation perfused with Krebs buffer solution, digoxin failed to produce vasoconstriction when administered intravenously (60 micrograms/kg) or in the perfusate at a concentration of 8 ng/ml, the blood level at the peak of the pressor response. Sodium-pump activity (ouabain-sensitive 86Rb+ uptake) of intralobular pulmonary arteries excised after 90 min of exposure to digoxin was the same as activity in arteries from control dogs. In conclusion, digoxin produces a pulmonary vasoconstriction through an alpha-adrenergic mechanism. Since the pressor response was observed only in the blood-perfused lung, blood-borne catecholamines are apparently involved.


Tumor exosomes inhibit differentiation of bone marrow dendritic cells.
The production of exosomes by tumor cells has been implicated in tumor-associated immune suppression. In this study, we show that, in mice, exosomes produced by TS/A murine mammary tumor cells target CD11b(+) myeloid precursors in the bone marrow (BM) in vivo, and that this is associated with an accumulation of myeloid precursors in the spleen. Moreover, we demonstrate that TS/A exosomes block the differentiation of murine myeloid precursor cells into dendritic cells (DC) in vitro. Addition of tumor exosomes at day 0 led to a significant block of differentiation into DC, whereas addition at later time points was less effective. Similarly, exosomes produced by human breast tumor cells inhibited the differentiation of human monocytes in vitro. The levels of IL-6 and phosphorylated Stat3 were elevated 12 h after the tumor exosome stimulation of murine myeloid precursors, and tumor exosomes were less effective in inhibiting differentiation of BM cells isolated from IL-6 knockout mice. Addition of a rIL-6 to the IL-6 knockout BM cell culture restored the tumor exosome-mediated inhibition of DC differentiation. These data suggest that tumor exosome-mediated induction of IL-6 plays a role in blocking BM DC differentiation.


Resveratrol alleviates bleomycin-induced lung injury in rats.
Antioxidant therapy may be useful in diseases with impaired oxidant-antioxidant balance such as pulmonary fibrosis. This study was designed to examine the effects of resveratrol, an antioxidant agents, against bleomycin-induced pulmonary fibrosis and oxidative damage. Wistar albino rats were administered a single dose of bleomycin (5 mg/kg; via the tracheal cannula) followed by either saline or resveratrol (10 mg/kg; orally) for 14 days. The effect of resveratrol on pulmonary oxidative damage was studied by cell count and analysis of cytokine levels (TGF-beta, TNF-alpha, IL-1beta and IL-6) in the bronchoalveolar lavage fluid (BALF) and biochemical measurements of malondialdehyde (MDA), an end product of lipid peroxidation; glutathione (GSH), a key antioxidant; and myeloperoxidase (MPO) activity, an index of neutrophil infiltration, in the lung tissue. Bleomycin-induced lung fibrosis was determined by lung collagen contents and also microscopically. Bleomycin caused a significant decrease in lung GSH, which was accompanied with significant increases in MDA level, MPO activity, and collagen contents of the lung tissue concomitant with increased levels of the pro-inflammatory mediators and cell count in BALF. On the other hand, resveratrol treatment reversed all these biochemical indices as well as histopathological alterations induced by bleomycin. The results demonstrate the role of oxidative mechanisms in bleomycin-induced pulmonary fibrosis, and resveratrol, by its antioxidant properties, ameliorates oxidative injury and fibrosis due to bleomycin. Thus, an effective supplement with resveratrol as an adjuvant therapy may be a very promising agent in alleviating the side effects of bleomycin, an effective chemotherapeutic agent.


Removal of ureteral calculi in two geldings via a standing flank approach.
Two geldings, aged 11 and 17 years, were examined for treatment of ureteroliths located approximately 10 cm proximal to the bladder. Ureteral obstruction was an incidental finding in 1 horse that was referred because of urinary tract obstruction and a cystic calculus. This horse did not have clinical or laboratory evidence of renal failure, although severe hydronephrosis was evident on transabdominal ultrasonography. The second patient had a serum creatinine concentration of 6.3 mg/dL (reference range, 0.8 to 2.2 mg/dL) and mild hydronephrosis of the affected left kidney. In both patients, the obstructed ureter was exteriorized through a flank incision as a standing procedure, and the calculus was crushed and removed with a uterine biopsy forceps introduced through a ureterotomy approximately 25 cm proximal to the calculus. The cystic calculus was removed through a perineal urethrostomy by lithotripsy, piecemeal extraction, and lavage. The horse without azotemia developed pyelonephritis in the affected kidney and was euthanatized because of complications of a nephrectomy 13 months later. In the horse with azotemia, the serum creatinine concentration decreased after surgery, and the horse returned to its intended use. However, it was euthanatized approximately 2 years after surgery because of progressive renal failure, and a large nephrolith was found in the previously unobstructed right kidney. The technique used for ureterolith removal was successful in both horses in this report, did not require sophisticated equipment, and could be effective in the early stages of ureteral obstruction as a means of restoring urine flow and renal function. The outcome in the horse with advanced unilateral renal disease without azotemia would suggest that nephrectomy should be considered as a treatment in such patients.


Whole Genome Expression Analysis in a Mouse Model of Tauopathy Identifies MECP2 as a Possible Regulator of Tau Pathology.
Increasing evidence suggests that hyperphosphorylation and aggregation of microtubule-associated protein tau (MAPT or tau) correlates with the development of cognitive impairment in Alzheimer's disease (AD) and related tauopathies. While numerous attempts have been made to model AD-relevant tau pathology in various animal models, there has been very limited success for these models to fully recapitulate the progression of disease as seen in human tauopathies. Here, we performed whole genome gene expression in a genomic mouse model of tauopathy that expressed human MAPT gene under the control of endogenous human MAPT promoter and also were complete knockout for endogenous mouse tau [referred to as 'hTau MaptKO(Duke)' mice]. First, whole genome expression analysis revealed 64 genes, which were differentially expressed (32 up-regulated and 32 down-regulated) in the hippocampus of 6-month-old hTau MaptKO(Duke) mice compared to age-matched non-transgenic controls. Genes relevant to neuronal function or neurological disease include up-regulated genes: PKC-alpha (Prkca), MECP2 (Mecp2), STRN4 (Strn4), SLC40a1 (Slc40a1), POLD2 (Pold2), PCSK2 (Pcsk2), and down-regulated genes: KRT12 (Krt12), LASS1 (Cers1), PLAT (Plat), and NRXN1 (Nrxn1). Second, network analysis suggested anatomical structure development, cellular metabolic process, cell death, signal transduction, and stress response were significantly altered biological processes in the hTau MaptKO(Duke) mice as compared to age-matched non-transgenic controls. Further characterization of a sub-group of significantly altered genes revealed elevated phosphorylation of MECP2 (methyl-CpG-binding protein-2), which binds to methylated CpGs and associates with chromatin, in hTau MaptKO(Duke) mice compared to age-matched controls. Third, phoshpho-MECP2 was elevated in autopsy brain samples from human AD compared to healthy controls. Finally, siRNA-mediated knockdown of MECP2 in human tau expressing N2a cells resulted in a significant decrease in total and phosphorylated tau. Together, these results suggest that MECP2 is a potential novel regulator of tau pathology relevant to AD and tauopathies.


Comparing employer-sponsored and federal exchange plans: wide variations in cost sharing for prescription drugs.
Just under seven million Americans acquired private insurance through the new health insurance exchanges, or Marketplaces, in 2014. The exchange plans are required to cover essential health benefits, including prescription drugs. However, the generosity of prescription drug coverage in the plans has not been well described. Our primary objective was to examine the variability in drug coverage in the exchanges across plan types (health maintenance organization or preferred provider organization) and metal tiers (bronze, silver, gold, and platinum). Our secondary objective was to compare the exchange coverage to employer-sponsored coverage. Analyzing prescription drug benefit design data for the federally facilitated exchanges, we found wide variation in enrollees' out-of-pocket costs for generic, preferred brand-name, nonpreferred brand-name, and specialty drugs, not only across metal tiers but also within those tiers across plan types. Compared to employer-sponsored plans, exchange plans generally had lower premiums but provided less generous drug coverage. However, for low-income enrollees who are eligible for cost-sharing subsidies, the exchange plans may be more comparable to employer-based coverage. Policies and programs to assist consumers in matching their prescription drug needs with a plan's benefit design may improve the financial protection for the newly insured.


Optical fiber-coupled ocular spectrometer for measurement of drug concentration in the anterior eye--applications in pharmaceuticals research.
This paper describes in detail a novel optoelectronic system designed to measure drug absorption in the anterior segment of the eye following topical application of drug formulations. This minimally invasive measurement technique offers both a method for determining drug concentration in human eyes, and demonstrates an alternative to current testing processes in model animals, which require paracentesis of the anterior chamber of the eye. The optoelectronic technique can be used with formulations, which possess appropriate spectral characteristics, namely unique absorption or fluorescence spectra. Preliminary experiments using our measurement system have been performed in rabbit and man, where we have been successful in achieving the direct measurement of topically applied brimonidine, an alpha-2 agonist used in the treatment of glaucoma. This demonstrates the feasibility of performing real-time, in vivo testing of ophthalmic drug formulations in the eye of human test subjects. We further demonstrate the novel application of the optoelectronic system for detection of topically applied UV-absorbing compounds in rabbit cadaver eyes, with a view to evaluating potential ocular sunscreen formulations. In summary, this method can be applied for the rapid comparison of the penetration of different drug formulations into the anterior eye at greatly reduced cost and time.


A 2015 global update on folic acid-preventable spina bifida and anencephaly.
Spina bifida and anencephaly are two major neural tube defects. They contribute substantially to perinatal, neonatal, infant, and under-five mortality and life-long disability. To monitor the progress toward the total prevention of folic acid-preventable spina bifida and anencephaly (FAP SBA), we examined their global status in 2015. Based on existing data, we modeled the proportion of FAP SBA that are prevented in the year 2015 through mandatory folic acid fortification globally. We included only those countries with mandatory fortification that added at least 1.0 ppm folic acid as a fortificant to wheat and maize flour, and had complete information on coverage. Our model assumed mandatory folic acid fortification at 200 μg/day is fully protective against FAP SBA, and reduces the rate of spina bifida and anencephaly to a minimum of 0.5 per 1000 births. Our estimates show that, in 2015, 13.2% (35,500 of approximately 268,700 global cases) of FAP SBA were prevented in 58 countries through mandatory folic acid fortification of wheat and maize flour. Most countries in Europe, Africa, and Asia were not implementing mandatory fortification with folic acid. Knowledge that folic acid prevents spina bifida and anencephaly has existed for 25 years, yet only a small fraction of FAP SBA is being prevented worldwide. Several countries still have 5- to 20-fold epidemics of FAP SBA. Implementation of mandatory fortification with folic acid offers governments a proven and rapid way to prevent FAP SBA-associated disability and mortality, and to help achieve health-related Sustainable Development Goals. Birth Defects Research (Part A) 106:520-529, 2016. © 2016 Wiley Periodicals, Inc.


HIV testing, human rights, and global AIDS policy: exceptionalism and its discontents.
Two years ago, in May 2007, UNAIDS and WHO issued new guidelines on HIV testing. Prepared to meet the demands of the AIDS pandemic and the prospects of extending the benefits of antiretroviral therapy to regions where such treatment had been all but out of reach, the new guidance was the product of an extended period of sometimes acrimonious controversy both within the two UN agencies and globally. Those pressing for change had argued that a paradigm of testing that had emerged at a time when little could be done for those infected with HIV was inappropriate to the current moment. Those who viewed with skepticism, if not hostility, the claims that current practice and stringent ethical standards had become an impediment to effectively confronting the challenge of AIDS saw in the proposed changes a threat to the bedrock ethical principles of informed consent. In the end, of course, decisions about HIV testing will be taken by nation - states, with the recommendations of international organizations constituting but one element, however important, that will shape policy. Nevertheless, an examination of the history and the dynamics of the recent controversy and its outcome will provide a unique resource to those faced with policy choices; it will also provide a unique opportunity to lay bare the complex and politically charged relationships evolving between public health and human rights.


A cholesterol-rich diet accelerates bacteriologic sterilization in pulmonary tuberculosis.
Hypocholesterolemia is common among tuberculous patients and is associated with mortality in miliary cases. Some in vitro studies have shown that cholesterol is necessary for the good functioning of macrophages and lymphocytes. To determine whether a cholesterol-rich diet could accelerate sputum sterilization in patients with pulmonary tuberculosis. An 8-week follow-up, randomized, controlled trial carried out from March 2001 to January 2002. A third-level hospital for respiratory diseases in Mexico City. Adult patients with newly diagnosed pulmonary tuberculosis were hospitalized for 8 weeks and randomly assigned to receive a cholesterol-rich diet (800 mg/d cholesterol [experimental group]) or a normal diet (250 mg/d cholesterol [control group]). All patients received the same four-drug antitubercular regimen (ie, isoniazid, rifampin, pyrazinamide, and ethambutol). Every week, a quantitative sputum culture and laboratory tests were done and respiratory symptoms were recorded. Patients in the experimental group (10 patients) and the control group (11 subjects) were HIV-negative and harbored Mycobacterium tuberculosis that was fully sensitive to antitubercular drugs. Sterilization of the sputum culture was achieved faster in the experimental group, as demonstrated either by the percentage of negative culture findings in week 2 (80%; control group, 9%; p = 0.0019) or by the Gehan-Breslow test for Kaplan-Meier curves (p = 0.0037). Likewise, the bacillary population decreased faster (p = 0.0002) in the experimental group. Respiratory symptoms improved in both groups, but sputum production decreased faster in the experimental group (p < 0.05). Laboratory test results did not differ between the groups. A cholesterol-rich diet accelerated the sterilization rate of sputum cultures in pulmonary tuberculosis patients, suggesting that cholesterol should be used as a complementary measure in antitubercular treatment.


Developmental differences in the accumbal dopaminergic response to repeated ethanol exposure.
Recent research indicates that alcohol use/abuse is often initiated during the adolescent period and that brain reinforcement pathways (e.g., the mesolimbic dopamine [DA] pathway) are undergoing developmental transition. Our research focuses on the effects of ethanol administration on neural mechanisms associated with addiction in preadolescent (postnatal day [PND] 25), adolescent (PND 35, PND 45), and young adult (PND 60) animals. Using conditioned place preference (CPP) testing, we have shown that adolescent animals are unique in their responses to ethanol. Since CPP has been associated with contextually conditioned incentive motivation, our results suggest that younger animals may be more vulnerable to addiction. The present data reveal that adolescent animals are neurochemically distinct in response to ethanol's effects. Using in vivo microdialysis within the nucleus accumbens septi (NAcc), we have determined the DAergic response across development. Results reveal that basal levels of DA transition during the adolescent period and differ from preadolescent or adult animals. Specifically, PND 45 animals exhibited significantly higher, and PND 25 significantly lower, basal DA levels than all other ages examined. Further, repeated exposure to ethanol elevated basal DA levels significantly regardless of age or dose. Basal 3,4-dihydroxyphenylacetic acid (DOPAC)/DA ratio also differed as a function of age, with PND 35 and PND 60 animals demonstrating the highest ratios, and PND 45 animals producing the lowest baseline levels. Repeated ethanol exposure produced significant changes in basal ratios as a function of age. Interestingly, PND 45 animals exhibited no change in ratios with repeated exposure, while all other ages demonstrated a dose-dependent rise in DOPAC/DA ratios. These data indicate an age-dependent difference in the homeostatic alterations of mesolimbic systems in response to repeated ethanol treatment, an effect that may manifest itself as differences in behavioral responsivity and conditionability to the drug and the drug's effects.


Efficacy of Paromomycin-Chloroquine Combination Therapy in Experimental Cutaneous Leishmaniasis.
The 4-aminoquinoline chloroquine (CQ) is clinically used in combination with doxycycline to cure chronic Q fever, as it enhances the activity of the antibiotic against the causative bacterium Coxiella burnetii residing within macrophage phagolysosomes. As there is a similar cellular host-pathogen biology for Leishmania parasites, this study aimed to determine whether such an approach could also be the basis for a new, improved treatment for cutaneous leishmaniasis (CL). We have evaluated the in vitro and in vivo activities of combinations of CQ with the standard drugs paromomycin (PM), miltefosine, and amphotericin B against Leishmania major and Leishmania mexicana In 72-h intracellular antileishmanial assays, outcomes were variable for different drugs. Significantly, the addition of 10 μM CQ to PM reduced 50% effective concentrations (EC50s) by over 5-fold against L. major and against normally insensitive L. mexicana parasites. In murine models of L. major and L. mexicana CL, daily coadministration of 50 mg/kg of body weight PM and 25 mg/kg CQ for 10 days resulted in a significant reduction in lesion size but not in parasite load compared to those for mice given the same doses of PM alone. Overall, our data indicate that PM-CQ combination therapy is unlikely to be a potential candidate for further preclinical development.


Efficient Semi-Numerical Implementation of Relativistic Exact Exchange within the Infinite-Order Two-Component Method Using a Modified Chain-of-Spheres Method.
We present an efficient implementation of relativistic exact exchange within the infinite-order two-component method (IOTC) by employing a state-of-the-art seminumerical integration technique. For accurate consideration of the picture change, inherent to two-component methods, we propose a new scheme based on a relativistic or picture-change transformation of the density matrix, which provides a simple and efficient formulation of relativistically transformed quantities such as the electron density or exact exchange and thus avoids expensive integral transformations. We show that the new scheme does not introduce additional numerical or theoretical errors beyond the approximations of the IOTC method. For the efficient implementation of exact-exchange integrals, we build upon a modified version of the chain-of-spheres exact-exchange (COSX) method. In addition to the conventional overlap and density matrix screening by S- and P-junctions, respectively, we introduce a new simple screening technique in the sense of the original COSX method by additionally considering the asymptotic decay of the integrals over the Coulomb operator within the new F-junctions. Together with the picture-change transformation of the density matrix, this modified COSX method is shown to provide superior efficiency for the calculation of relativistic exact exchange compared to a conventional analytical direct self-consistent-field implementation of exact exchange.


Cloning of Drosophila MCM homologs and analysis of their requirement during embryogenesis.
MCM (minichromosome maintenance) gene family of Saccharomyces cerevisiae encodes essential DNA replication factors that participate in the initiation of DNA replication. In addition, their localization to the nucleus in a mitosis-dependent manner fueled the hypothesis that MCMs also act to couple DNA replication to mitosis. We report the identification of a Drosophila gene family with extensive sequence identity to the MCM genes. Results from antibody injection experiments suggest that MCMs play an essential role in DNA replication during embryogenesis. Evolutionary conservation of MCM sequences and function in Drosophila could potentially facilitate studies of how initiation of DNA replication is regulated and coupled to mitosis during metazoan development.


Selective inhibition of matrix metalloproteinase 10 (MMP10) with a single-domain antibody.
Since their discovery, the matrix metalloproteinase (MMP) family proteases have been considered as therapeutic targets in numerous diseases and disorders. Unfortunately, clinical trials with MMP inhibitors have failed to yield any clinical benefits of these inhibitors. These failures were largely due to a lack of MMP-selective agents; accordingly, it has become important to identify a platform with which high selectivity can be achieved. To this end, we propose using MMP-targeting antibodies that can achieve high specificity in interactions with their targets. Using a scaffold of single-domain antibodies, here we raised a panel of MMP10-selective antibodies through immunization of llamas, a member of the camelid family, whose members generate conventional heavy/light-chain antibodies and also smaller antibodies lacking light-chain and CH1 domains. We report the generation of a highly selective and tightly binding MMP10 inhibitor (Ki < 2 nm). Using bio-layer interferometry-based binding assays, we found that this antibody interacts with the MMP10 active site. Activity assays demonstrated that the antibody selectively inhibits MMP10 over its closest relative, MMP3. The ability of a single-domain antibody to discriminate between the most conserved MMP pair via an active site-directed mechanism of inhibition reported here supports the potential of this antibody as a broadly applicable scaffold for the development of selective, tightly binding MMP inhibitors.


Survival of rabbit limbal stem cell allografts.
Failure of a specialized population of corneal epithelial stem cells found in the peripheral cornea and limbus results in ocular surface disease, which may be amenable to treatment by transplantation of limbal tissue. This study was designed to investigate donor limbal stem cell allograft survival in rabbits with ocular surface disease. Rabbits underwent corneal epithelial debridement and limbal ablation to induce ocular surface disease and were then treated by limbal stem cell allotransplantation, by allotransplantation plus topical steroid, or by topical steroid only (n = 7 for each group). Donors and recipients were sex mismatched. Recipients were followed for up to 5 months. Outcome was assessed by daily slit-lamp examination, weekly impression cytology and photographic record, end-point sex chromatin and fluorescent cell tracer analyses, histology, and immunohistochemistry. In no case was a completely normal ocular surface regained, but some animals that received grafts plus corticosteroids fared best by all criteria used. In the absence of immunosuppression, graft hemorrhagia (believed to be a manifestation of graft rejection) occurred within the first month, the cornea became resurfaced with conjunctiva-derived cells, and no donor cells survived centrally in the long term. Topical corticosteroids reduced the number and severity of these episodes significantly, and were associated with survival of some donor-derived cells in the central cornea of some grafted animals. Thus, rabbit limbal stem cell allografts appeared to undergo rejection, which could be modified by immunosuppression, but useful regeneration of the ocular surface occurred only where rejection was circumvented.


Information-Theoretic Model and Analysis of Molecular Signaling in Targeted Drug Delivery.
Targeted drug delivery (TDD) modality promises a smart localization of appropriate dose of therapeutic drugs to the targeted part of the body at reduced system toxicity. To achieve the desired goals of TDD, accurate analysis of the system is important. Recent advances in molecular communication (MC) present prospects to analyzing the TDD process using engineering concepts and tools. Specifically, the MC platform supports the abstraction of TDD process as a communication engineering problem in which the injection and transportation of drug particles in the human body and the delivery to a specific tissue or organ can be analyzed using communication engineering tools. In this paper we stand on the MC platform to present the information-theoretic model and analysis of the TDD systems. We present a modular structure of the TDD system and the probabilistic models of the MC-abstracted modules in an intuitive manner. Simulated results of information-theoretic measures such as the mutual information are employed to analyze the performance of the TDD system. Results indicate that uncertainties in drug injection/release systems, nanoparticles propagation channel and nanoreceiver systems influence the mutual information of the system, which is relative to the system's bioequivalence measure.


Development of a novel transgenic mouse for the study of interactions between CD4 and CD8 T cells during graft rejection.
The goal of this study was the development of a system in which the cooperative interactions between CD4 and CD8 T cells specific for defined peptides from a single minor histocompatibility antigen could be studied. A transgenic mouse strain that expresses chicken ovalbumin (Act-mOVA) on the surface of all cells in the body was produced as a source of tissues containing such an antigen. Skin grafts from Act-mOVA donors were rapidly and completely rejected by wild-type recipients, but only when both CD4 and CD8 T cells were present. CD4 T cells by themselves caused an incomplete form of rejection characterized by rapid but partial contraction of Act-mOVA grafts. CD8 T cells alone caused complete rejection of Act-mOVA skin grafts but only after a long delay. Adoptively transferred ovalbumin-specific TCR-transgenic CD4 and CD8 T cells were stimulated by Act-mOVA graft antigens and CD8 T-cell accumulation in the grafts was enhanced by specific CD4 T cells. These findings, together with the fact that the ligand for ovalbumin peptide-specific CD8 T cells can be detected in Act-mOVA tissues with an MHC-restricted antibody, make this an ideal system for the study of cooperation between CD4 and CD8 T cells.


The use of matched molecular series networks for cross target structure activity relationship translation and potency prediction.
Matched molecular series (MMS) analysis is an extension of matched molecular pair (MMP) analysis where all of the MMPs belong to the same chemical series. An MMS within a biological assay is able to capture specific structure activity relationships resulting from chemical substitution at a single location in the molecule. Under this convention, an MMS has the ability to capture one specific interaction vector between the compounds in a series and their therapeutic target. MMS analysis has the potential to translate the SAR from one series to another even across different protein targets or assays. A significant limitation of this approach is the lack of chemical series with a sufficient number of overlapping fragments to establish a statistically strong SAR in most databases. This results in either an inability to perform MMS analysis altogether or a potentially high proportion of spurious matches from chance correlations when the MMS compound count is low. This paper presents the novel concept of an MMS Network, which captures the SAR relationships between a set of related MMSs and significantly enhances the performance of MMS analysis by reducing the number of spurious matches leading to the identification of unexpected and potentially transferable SAR across assays. The results of a full retrospective leave-one-out analysis and randomization simulation are provided, and examples of pharmaceutically relevant programs will be presented to demonstrate the potential of this method.


Two-Dimensional, High-Resolution Peripheral Refraction in Adults with Isomyopia and Anisomyopia.
The purpose of this study was to investigate the two-dimensional peripheral refraction in fellow eyes of patients with isomyopia and anisomyopia. Sixty-eight young adults were recruited, including 25 isomyopes with interocular differences (IODs) of foveal refraction < 1.00 D and 43 anisomyopes with IOD > 1.50 D. Peripheral refraction across an area of the visual field of 60° × 36° with a resolution of 1° was measured using a custom-made Hartmann-Shack wavefront sensor. The retinal area was divided into 3 × 3 zones for comparison between the fellow eyes. There was no difference of refraction in all corresponding zones between the fellow eyes in the isomyopic group (all P > 0.05). The IODs between more myopic (MM) eyes and less myopic (LM) eyes in the anisomyopic group ranged from -1.40 to approximately -2.46 D (all P <0.001), which was flagged in the center and attenuated in peripheral zones by varied magnitudes. In the stratification analysis for different levels of anisomyopia, the nasal retina first presented significant relative hyperopic shifts compared to the center, followed by the temporal retina. In contrast, the superior and inferior periphery only differed from the center when the central IOD was greater than 3.00 D. The two-dimensional peripheral refraction patterns showed a mirror symmetry between the fellow eyes of a patient with isomyopia. However, in the anisomyopic group, peripheral refraction showed significantly relative hyperopic shift when compared with the center and developed with a varied rate in different areas. These findings may indicate an asymmetrical variation in the peripheral refraction patterns during myopia progression.


Smaller hippocampal volume as a vulnerability factor for the persistence of post-traumatic stress disorder.
Smaller hippocampal volume has often been observed in patients with post-traumatic stress disorder (PTSD). However, there is no consensus whether this is a result of stress/trauma exposure, or constitutes a vulnerability factor for the development of PTSD. Second, it is unclear whether hippocampal volume normalizes with successful treatment of PTSD, or whether a smaller hippocampus is a risk factor for the persistence of PTSD. Magnetic resonance imaging (MRI) scans and clinical interviews were collected from 47 war veterans with PTSD, 25 healthy war veterans (combat controls) and 25 healthy non-military controls. All veterans were scanned a second time with a 6- to 8-month interval, during which PTSD patients received trauma-focused therapy. Based on post-treatment PTSD symptoms, patients were divided into a PTSD group who was in remission (n = 22) and a group in whom PTSD symptoms persisted (n = 22). MRI data were analysed with Freesurfer. Smaller left hippocampal volume was observed in PTSD patients compared with both control groups. Hippocampal volume of the combat controls did not differ from healthy controls. Second, pre- and post-treatment analyses of the PTSD patients and combat controls revealed reduced (left) hippocampal volume only in the persistent patients at both time points. Importantly, hippocampal volume did not change with treatment. Our findings suggest that a smaller (left) hippocampus is not the result of stress/trauma exposure. Furthermore, hippocampal volume does not increase with successful treatment. Instead, we demonstrate for the first time that a smaller (left) hippocampus constitutes a risk factor for the persistence of PTSD.