Evidence of Antineutrinos from Distant Reactors Using Pure Water at SNO.

The SNO+ Collaboration reports the first evidence of reactor antineutrinos in a Cherenkov detector. The nearest nuclear reactors are located 240 km away in Ontario, Canada. This analysis uses events with energies lower than in any previous analysis with a large water Cherenkov detector. Two analytical methods are used to distinguish reactor antineutrinos from background events in 190 days of data and yield consistent evidence for antineutrinos with a combined significance of 3.5σ.

[Therapeutic effect of encapsulated into the nanocontainers MBP immunodominant peptides on EAE development in DA rats].

Multiple Sclerosis (MS) is a serve autoimmune neurodegenerative disease. Development of innovative approaches of MS treatment is of a high priority in the modern immunology and pharmacy. In the present study we showed high therapeutic efficiency of immunodominant peptides of myelin basic protein (MBP) incorporated into the monolayer mannosylated liposomes on the development of experimental autoimmune encephalomyelitis (EAE) in DA rats. MBP is a component ofoligodendrocytes' membrane, which form axonal sheath, and is one of the major autoantigens in MS. We analyzed binding pattern ofanti-MBP autoantibodies from MS patients using previously designed MBP epitope library. Utilizing the same approach we investigated pool of anti-MBP antibodies from SJL/J and C57/BL6 mice and DA rats with induced EAE. The most relevant rodent model to MS was EAE in DA rats according to the autoantibodies' binding pattern. We selected three immunodominant MBP fragments encapsulated in monolayer mannosylated liposomes for the following treatment of verified DA rodent model. MBP fragment 46-62 was the most effective in reducing of the first EAE attack, whereas MBP 124-139 and 147-160 inhibited development of pathology during remission stage. Simultaneous administration of these peptides in liposomes significantly decreased level of anti-MBP antibodies. Synergetic therapeutic effect of MBP fragments reduced integral disease score by inhibiting first EAE wave and subsequent remission, thus, our findings disclosure novel approaches for efficient treatment of Multiple Sclerosis.

Selective silencing of the hypoxia-inducible factor 1 target gene BNIP3 by histone deacetylation and methylation in colorectal cancer.

Hypoxia, via the hypoxia-inducible factors 1 and 2 (HIF-1 and HIF-2), upregulates many genes involved in cell survival. However, proapoptotic pathways are also induced. BCL-2/adenovirus E1B-19 kDa-interacting protein 3 (BNIP3) represents a paradigm of a cell death protein that is hypoxically upregulated via HIF-1 in most cancers. We found that in contrast to many other cell types, 6/8 colorectal cancer (CRC) cell lines show little hypoxic induction of BNIP3 despite an intact HIF signalling system. Colorectal tumour tissue also loses BNIP3 expression relative to matched normal samples. Downregulation of hypoxic BNIP3 in CRC cells was independent of the expression of other BCL-2 family members, or BNIP3L. That BNIP3 plays a functional role in hypoxic survival in CRC cells was demonstrated by the fact that CRC cell lines that do not upregulate BNIP3 or have been treated with BNIP3 RNA interference were insensitive to hypoxia-induced cell death. Promoter methylation and histone deacetylation were shown to silence BNIP3 in these CRC cell lines. Of significance, hypoxic induction of BNIP3 was restored in 4/6 cell lines by trichostatin-A treatment alone. These data suggest that BNIP3 plays an important role in hypoxic cell death and epigenetic mechanisms selectively silence its expression in CRC.

Evaluation of an adult immunization composite measure in the Indian Health Service.

BACKGROUND: Government agencies, healthcare accreditation bodies and quality improvement organizations support the development of new quality measures. Composite quality measures use more than one measure to develop a broader assessment of healthcare system function. Currently, no composite measures for adult immunization coverage exist. Development of such measures could facilitate improvements in adult immunization coverage by focusing on measurement of receipt of all age-recommended vaccines. METHODS: We recruited five Indian Health Service (IHS) and Tribal health clinics to pilot an Adult Immunization Composite Measure (AICM). Data were collected monthly over seven months using a pre-programmed electronic health record (EHR) reporting tool (IHS sites); Tribal sites used third-party software or a programmable EHR reporting function. Data collected included: number of adults aged 19 years and over who were active users of the facility with at least two visits in the last three years; the cumulative number fully immunized per age-based recommendations for tetanus toxoid-containing vaccines, pertussis, zoster and pneumococcal vaccines; and the percent immunized for the AICM and for each individual vaccine. Coverage was calculated for three age groups: 19-59 years; 60-64 years; and 65 years and older. RESULTS: All sites reported aggregate immunization data monthly from patient EHR records. For all adults 19 years and older, AICM coverage ranged from 49% to 87% at the end of the report period. Two sites showed increases in AICM coverage ≥ 3%. Improvements in zoster vaccine coverage accounted for most of the increase observed. One site specifically focused on improving zoster coverage as a result of using the AICM. CONCLUSIONS: We demonstrated the feasibility of implementing a composite measure of adult immunization coverage. This is the first measure capable of monitoring immunization completeness, coverage improvement and overall adult vaccine program effectiveness for adults who receive all recommended, age-based vaccines.

Elevated PTTG and PBF predicts poor patient outcome and modulates DNA damage response genes in thyroid cancer.

The proto-oncogene PTTG and its binding partner PBF have been widely studied in multiple cancer types, particularly thyroid and colorectal, but their combined role in tumourigenesis is uncharacterised. Here, we show for the first time that together PTTG and PBF significantly modulate DNA damage response (DDR) genes, including p53 target genes, required to maintain genomic integrity in thyroid cells. Critically, DDR genes were extensively repressed in primary thyrocytes from a bitransgenic murine model (Bi-Tg) of thyroid-specific PBF and PTTG overexpression. Irradiation exposure to amplify p53 levels further induced significant repression of DDR genes in Bi-Tg thyrocytes (P=2.4 × 10-4) compared with either PBF- (P=1.5 × 10-3) or PTTG-expressing thyrocytes (P=NS). Consistent with this, genetic instability was greatest in Bi-Tg thyrocytes with a mean genetic instability (GI) index of 35.8±2.6%, as well as significant induction of gross chromosomal aberrations in thyroidal TPC-1 cells following overexpression of PBF and PTTG. We extended our findings to human thyroid cancer using TCGA data sets (n=322) and found striking correlations with PBF and PTTG expression in well-characterised DDR gene panel RNA-seq data. In addition, genetic associations and transient transfection identified PBF as a downstream target of the receptor tyrosine kinase-BRAF signalling pathway, emphasising a role for PBF as a novel component in a pathway well described to drive neoplastic growth. We also showed that overall survival (P=1.91 × 10-5) and disease-free survival (P=4.9 × 10-5) was poorer for TCGA patients with elevated tumoural PBF/PTTG expression and mutationally activated BRAF. Together our findings indicate that PBF and PTTG have a critical role in promoting thyroid cancer that is predictive of poorer patient outcome.

Hypermutability at a poly(A/T) tract in the human germline.

Poly(A/T) tracts are abundant simple sequence repeats (SSRs) within the human genome. They constitute part of the coding sequence of a variety of genes, encoding polylysine stretches that are important for protein function. Assessment of poly(A/T) tract stability is also used to identify microsatellite unstable colorectal cancers, which are characteristic of tumours defective in DNA mismatch repair. Despite their importance, little is known about the stability of poly(A/T) SSRs in the human germline. We have determined the stability of a paradigm poly(A/T) tract, BAT-40, by study of population allele frequencies, mutation frequency in families and mutation frequency in sperm DNA. We show that the locus is polymorphic, with a level of heterozygosity of 59.7%. Germline mutation was observed in 13 of 187 germline transmissions (7.0%) in 10 families suggesting BAT-40 is unstable in the germline. Further evidence for germline instability at BAT-40 was provided by small pool PCR analysis of matched blood and sperm DNA templates, revealing a significantly elevated frequency of mutation in the germline (P < 0.001). These findings provide insight into poly(A/T) tract stability in the germline. They also have relevance to the study of gene expression and to determination of microsatellite instability in tumours.

Mutation frequency in coding and non-coding repeat sequences in mismatch repair deficient cells derived from normal human tissue.

Repetitive tracts within the coding regions of TGFBR2 and BAX are frequently mutated in mismatch repair deficient tumours and are implicated in tumour progression. However, there has been little study of the balance between selection pressure and inherent instability at sequences within these genes. To determine whether TGFBR2 and BAX are inherently prone to mutations in the presence of MMR defects, we studied MMR deficient cells derived from B-lymphocytes. By analysing cells derived from normal tissue we aimed to minimize the effects of selection pressures that bias the apparent frequency of mutation. We definitively show that certain sequences, usually repaired by MMR, are inherently unstable. Using a small pool PCR technique we confirmed these cells exhibit microsatellite instability. Additionally, we demonstrate that MMR deficiency results in an excess of mutations, specifically at the poly(A)(10) tract compared to other regions of the TGFBR2 gene (P<0.001). Conversely, an excess of mutations does not appear to arise at the poly(G)(8) tract of the BAX gene. These studies provide insight into the mechanism by which TGFBR2 and BAX genes become mutated during tumorigenesis. These findings invoke the notion of "unmasking" specific hypermutable sequences in particular genes adding further complexity to the concept of the mutator phenotype.

Blocking CLEC14A-MMRN2 binding inhibits sprouting angiogenesis and tumour growth.

We previously identified CLEC14A as a tumour endothelial marker. Here we show that CLEC14A is a regulator of sprouting angiogenesis in vitro and in vivo. Using a human umbilical vein endothelial cell spheroid-sprouting assay, we found CLEC14A to be a regulator of sprout initiation. Analysis of endothelial sprouting in aortic ring and in vivo subcutaneous sponge assays from clec14a(+/+) and clec14a(-/-) mice revealed defects in sprouting angiogenesis in CLEC14A-deficient animals. Tumour growth was retarded and vascularity reduced in clec14a(-/-) mice. Pull-down and co-immunoprecipitation experiments confirmed that MMRN2 binds to the extracellular region of CLEC14A. The CLEC14A-MMRN2 interaction was interrogated using mouse monoclonal antibodies. Monoclonal antibodies were screened for their ability to block this interaction. Clone C4, but not C2, blocked CLEC14A-MMRN2 binding. C4 antibody perturbed tube formation and endothelial sprouting in vitro and in vivo, with a similar phenotype to loss of CLEC14A. Significantly, tumour growth was impaired in C4-treated animals and vascular density was also reduced in the C4-treated group. We conclude that CLEC14A-MMRN2 binding has a role in inducing sprouting angiogenesis during tumour growth, which has the potential to be manipulated in future antiangiogenic therapy design.

Adhesion molecules and relationship to leukocyte levels in allergic eye disease.

PURPOSE: To evaluate the conjunctival expression of leukocyte cell adhesion molecules (CAMs) and their relationship to leukocyte patterns on the microvasculature in the different clinical subtypes of allergic eye disease. METHODS: Immunohistochemical analysis, using appropriate monoclonal antibodies, was applied to glycolmethacrylate-embedded biopsies of bulbar and tarsal conjunctival tissue. The proportion of total blood vessels expressing a particular CAM was derived and related to individual cell types identified by cell-specific markers, such as mast cells, eosinophils, neutrophils, T cells, and macrophages. Statistical analysis was used to correlate adhesion molecule expression and, ultimately, cell type. RESULTS: There was a basal expression of intercellular adhesion molecule-1 (ICAM-1) (21% bulbar, 18% tarsal), E-selectin (15% bulbar, 21% tarsal), and vascular cell adhesion molecule-1 (VCAM-1) (13% bulbar and tarsal) in normal controls. In seasonal and perennial (bulbar and tarsal conjunctival) allergic tissue, ICAM-1 and E-selectin were expressed in 40% to 78% of vessels; in chronic disease, they were expressed in 45% to 80% of vessels; and in vernal giant papillae, they were expressed in as many as 90% of vessels. There was also increased expression of endothelial VCAM-1 in all forms of allergic eye disease; the greatest values were found in vernal giant papillae (64%). Biopsies taken in winter from seasonal sufferers demonstrated a marked reduction in levels of all three CAMs compared with those taken in the pollen season. This is almost consistent with values found in normal conjunctiva. Positive correlations were found between the levels of ICAM-1 and E-selectin expression and the degree of granulocyte and lymphocyte infiltration, although VCAM-1 expression correlated most closely with eosinophil numbers. CONCLUSIONS: Increased levels of cell adhesion molecules on the microvasculature and the factors that regulate them are likely to be responsible for the infiltration of cells bearing their ligands and may perpetuate inflammation in the chronic forms of allergic eye disease.

Very early changes in olfactory functioning due to Alzheimer's disease and the role of apolipoprotein E in olfaction.

Alzheimer's disease (AD) is a progressive neurodegenerative illness marked by memory loss and at least one other cognitive disturbance. Early diagnosis of the disease has proved difficult and has therefore been the focus of much research. Apolipoprotein E (ApoE), a protein manufactured and distributed throughout the body, has shown specificity of binding to the beta A4 peptide, the primary component in the senile plaques of AD. Furthermore, the ApoE, epsilon 4 (epsilon 4) allele, is overrepresented in AD. These two lines of evidence suggest that ApoE, specifically the epsilon 4 allele, plays an important role in the development of AD. Further support for this hypothesis appears in neuropsychological data showing cognitive decrements in ostensibly nondemented individuals with the epsilon 4 allele, compared to those without the allele. It is also well known that olfaction is compromised in AD. Thus, the purpose of this study was twofold: (1) to examine very early changes in olfactory functioning due to AD and (2) to examine the role of ApoE in olfactory functioning in people at risk for AD by virtue of early cognitive decline. Results demonstrated changes in olfactory threshold the year immediately preceding change in diagnosis from normal control to AD. Also, in individuals with mild cognitive impairment, those with the ApoE epsilon 4 allele show poorer thresholds than those without the epsilon 4 allele.

Apolipoprotein E status is associated with odor identification deficits in nondemented older persons.

Alzheimer's disease (AD) patients with moderate dementia show losses in olfactory threshold, odor identification and odor memory. Sensitivity and specificity of olfactory testing is significant, with the greatest power of accurate diagnosis in the more cognitively loaded olfactory tasks. In patients with very mild AD or in patients at risk for the disease because of their mild cognitive impairment, losses are apparent for odor identification, odor recognition memory and odor threshold, with the best sensitivity in the identification task. Persons who are either heterozygous or homozygous for the epsilon 4 allele of apolipoprotein E (ApoE) have an increased risk of Alzheimer's disease, although they show no dementia in the preclinical period. Evidence of olfactory dysfunction in this population might be reflective of an incipient dementing process. We have recently examined olfactory function in a group of normal elderly persons who have undergone genetic testing for the Apoe4 allele. These individuals consisted of all normal control subjects at the University of California, San Diego (UCSD) Alzheimer's Disease Research Center (ADRC) who had undergone both the genetic testing and testing for olfactory function. All had been diagnosed as normal control participants by two different neurologists who applied the National Institute of Neurological Disorders and Stroke and the Alzheimer's Disease and Related Disorders Association (NINDS-ADRDA) criteria for dementia. Persons with a history of alcoholism, drug abuse, learning disability or neurologic or psychiatric illness (including depression) were excluded. In this population, persons with the Apoe4 allele showed significantly poorer odor identification than those without an epsilon 4 allele. Early appearance of olfactory deficits in the progression to AD in persons with the epsilon 4 allele suggests diagnostic utility in olfactory testing.

Immunoglobulin G directed against toxins A and B of Clostridium difficile in the general population and patients with antibiotic-associated diarrhea.

Serum immunoglobulin G (IgG) class antibodies directed against toxins A and B of Clostridium difficile were studied using an enzyme-linked immunosorbent assay and a serum-neutralizing assay based on the MRC-5 tissue cytotoxicity assay. Of 185 individuals, 46 sera (24%) in the general population demonstrated IgG antibody, 36 (19.4%) against toxin A and 15 (8.1%) against toxin B. Antibody titer in the general population did not correlate with serum-neutralizing activity. Antibody prevalence fell with age (P = 0.58) over 50 years. Six of ten patients with acute primary episodes of C. difficile-associated diarrhea demonstrated antibody in convalescent-phase sera, predominantly directed against toxin B. Only two (28%) of seven patients with a history of relapsing C. difficile disease had demonstrable antibody.

Targeted disruption of galanin: new insights from knock-out studies.

The neuropeptide galanin has a widespread but no means ubiquitous expression pattern in the nervous and endocrine systems. Profound changes in the levels and distribution of the peptide occur in a range of path-physiological situations including nerve injury or damage and alterations in the circulating levels of a number of hormones. There is now a substantial body of work to indicate that galanin plays an important biological role as a regulator of neurotransmitter and hormone release in the adult. The recent generation of mice carrying a loss-of-function mutation within the galanin gene has allowed us new insights into the physiological actions of galanin. In this manuscript we detail three sets of data relating to the major phenotypic effects thus far delineated, putting them in the context of existing published data. These studies demonstrate that galanin acts as a developmental and trophic factor to subsets of neurons in the nervous and neuroendocrine systems.

Perfluorocarbon heavy liquids in the management of posterior dislocation of the lens nucleus during phakoemulsification.

We report a case in which the lens nucleus dislocated into the vitreous cavity through a posterior capsular rupture during phakoemulsification. We performed a vitrectomy and removed the lens nucleus using the perfluorocarbon heavy liquid perfluoro-1,3-dimethylcyclohexane. The management of posterior dislocation of the lens nucleus during cataract surgery is discussed.

Effect of race on perception of fat alone and in combination with sugar.

Two studies were performed to assess the perception of sugar-fat combinations and fat emulsions in African-American and white subjects. In the first study, African-American children aged 9-15 years were found to prefer higher concentrations of sweetness in liquid dairy products varying in fat content than white children. No significant differences in preference for the four fat levels were found. These data are consistent with a previous study by Desor et al. (2) that suggested African-American youngsters aged 9-15 preferred greater sweetness in water solutions. In a second study, thresholds and preferences for corn oil and butterfat in emulsions were determined for young adults. No significant differences between African-American and white young adults were found.

The responses of autistic children to the distress of others.

The behavior of preschool children from five groups (developmental language disordered, high-functioning autistic, low-functioning autistic, mentally retarded, and normally developing) were coded in three situations: presentation of a nonsocial orienting stimulus (an unfamiliar noise) and two social situations involving simulated distress on the part of an adult with whom they were playing. Cognitive level was correlated with level of responsiveness to stimuli only for the two retarded groups (mentally retarded and low-functioning autistic). Girls showed more prosocial behavior than boys in both social situations, independent of diagnosis. The language-disordered children showed only mild and subtle social deficits. The low-functioning autistic children showed pronounced deficits in responding in all situations. The mentally retarded and high-functioning autistic children showed good awareness of all situations, but were moderately impaired in their ability to respond prosocially; they rarely initiated prosocial behavior, but did respond to specific prompts. The behavioral feature that marked both autistic groups, in contrast to all other groups, was a lack of social referencing; they did not tend to look toward an adult in the presence of an ambiguous and unfamiliar stimulus. Results are discussed in terms of variability between and among high- and low-functioning autistic children, and implications for the core deficits in autism.

Silicone rubber contact lenses for the compromised cornea.

Silicone rubber contact lenses (SRCLs) are infrequently used because of the risk of developing unpredictable lens tightening, their poor availability, and their expense. However, their high oxygen transmissibility and nonabsorption of water make them valuable as therapeutic lenses. SRCLs are routinely used in our management of severely dry eyes, decompensated or vascularised corneas, and conditions where the corneal shape is flat or irregular. The records of 48 consecutive patients fitted with SRCLs between January 1989 and June 1990 were studied. The clinical history, indications, complications, success, and duration of SRCL wear were analysed. Therapeutic goals, which included epithelial healing, sealing of corneal perforations, and improved comfort and vision, were achieved in 53 of 62 eyes. The best corrected acuity was attained using SRCLs in 58 of 62 eyes. Failure of lens wear was due to lens tightening (four eyes), spoilation (two), discomfort, fornix shortening, handling problems, and decentration (one each). Infective keratitis complicated one case, but SRCL wear was resumed after successful treatment. With adequate follow-up, SRCLs have a low complication rate and are well tolerated even in severely compromised eyes, for which conventional lenses may be contraindicated. Their continued use as therapeutic lenses is advocated in carefully selected cases.

The importance of the short-term leaching dynamics of wood preservatives.

The potential environmental impacts from the use of treated timber in aquatic areas is under scrutiny as a result of environmental legislation and reports of the deleterious environmental effects around treated structures. In this study leaching experiments of up to 3 weeks duration were conducted on two species of chromated copper arsenate treated timber, dried for different periods of time. Increased drying time significantly reduced leaching of Cr and As. The addition of a synthetic humic acid increased leaching of Cu and As, but reduced leaching of Cr. Putative risk assessments conducted using short-term copper leaching data suggested protocol design may influence decisions made regarding the environmental acceptability of such preservatives.

Risk factors for acanthamoeba keratitis in contact lens users: a case-control study.

OBJECTIVE: To investigate reasons for an increase in cases of Acanthamoeba keratitis related to contact lenses. DESIGN: Case-control study. Cases were contact lens related acanthamoeba keratitis patients treated between 1 September 1989 and 31 August 1992. Controls were lens users without lens related disease who presented as new patients to the casualty department from 1 March 1992 to 31 August 1992. All subjects completed a questionnaire detailing lens use and hygiene practices. SETTING: Eye hospital. SUBJECTS: 35 cases with acanthamoeba keratitis and 378 controls. MAIN OUTCOME MEASURES: Relative risks comparing different contact lens types, socioeconomic classification, age, sex, lens use, lens wearing experience, hygiene compliance, and hygiene systems. RESULTS: The crude relative risk for developing acanthamoeba keratitis with the use of daily wear disposable lenses was 49.45 (95% confidence interval 6.53 to 2227; P < 0.001) compared with conventional soft lenses (the referent). Multivariable analysis showed that this increased risk could be largely attributed to lack of disinfection (relative risk 55.86 (10 to 302); P < 0.001) and use of chlorine based disinfection (14.63 (2.8 to 76); P = 0.001) compared with other chemical systems (the referent). None of the other outcome measures showed a significant association. CONCLUSIONS: Both failure to disinfect daily wear soft contact lenses and the use of chlorine release lens disinfection systems, which have little protective effect against the organism, are major risk factors for acanthamoeba keratitis. These risks have been particularly common in disposable lens use. Over 80% of acanthamoeba keratitis could be avoided by the use of lens disinfection systems that are effective against the organism.