Comparing Parent and Teacher Ratings of Emotional and Behavioural Difficulties in 5-year old Children who are Deaf or Hard-of-Hearing.

Children who are deaf or hard of hearing (DHH) are at higher risk of developing mental health problems. This study reports on the parent and teacher ratings of emotional and behavioural difficulties (EBD) in 5-year old DHH children. It explores the similarities and differences between informants, and the risk and protective factors associated with parent and teacher-ratings of EBD. Parents and teachers of 224 DHH children completed questionnaires on children's EBD and functional auditory behaviour. Children completed standardised assessments of non-verbal cognitive and language abilities. On average, parent- and teacher-rated EBD were 0.42 and 0.20 standard deviations higher than typically developing children. Parents reported more behavioural problems (hyperactivity and conduct), whereas teachers reported poorer prosocial behaviour. Inter-rater correlations were generally low to moderate (0.29 to 0.50). Overall, children with additional disabilities, lower non-verbal cognitive ability, and poor functional auditory behaviour were at higher risk of EBD. Language ability was only a significant predictor of teacher-rated EBD for children with hearing aids but not cochlear implants. Differences in informant-ratings emphasize the need for a multi-informant approach to get a global perspective on the psychopathology of DHH children. The findings indicate that parents may need assistance with managing behavioural problems at home, and teachers should facilitate more opportunities to practice prosocial behaviour at school. Intervention efforts should focus on facilitating good functional listening skills, as this may in turn, improve the mental health of young DHH children.

Language development in deaf or hard-of-hearing children with additional disabilities: type matters!

BACKGROUND: This study examined language development in young children with hearing loss and different types of additional disabilities (ADs). METHOD: A population-based cohort of 67 children who were enrolled in the Longitudinal Outcomes of Children with Hearing Impairment study took part. Language ability was directly assessed at 3 and 5 years of age using the Preschool Language Scale, Fourth Edition and the Peabody Picture Vocabulary Test, Fourth Edition. Standard scores were used to enable comparison with age-based expectations for typically developing children. RESULTS: Analysis of variance showed that, across the total cohort, children's language scores remained stable over the 2-year period. However, this overall stability masked a significant difference between children with different types of ADs; in particular, children with autism, cerebral palsy and/or developmental delay showed a decline in standard scores, whereas children with other disabilities showed a relative improvement. In addition, larger improvements in receptive vocabulary were associated with use of oral communication only. CONCLUSIONS: The results suggest that type of AD can be used to gauge expected language development in the population of children with hearing loss and ADs when formal assessment of cognitive ability is not feasible.

The effect of pre-analytical variables on downstream application and data analysis of human endometrial biopsies.

STUDY QUESTION: What are the effects of pre-analytical variables on the downstream analysis of patient-derived endometrial biopsies? SUMMARY ANSWER: There are distinct differences in the protein levels of the master regulator of oxygen homeostasis, hypoxia-inducible factor-1-alpha (HIF1α), and the protein and mRNA levels of three related genes, carbonic anhydrase 9 (CA9), vascular endothelial growth factor A (VEGFA) and progesterone receptor (PR) in human endometrial biopsies, depending on the pre-analytical variables: disease status (cancer vs benign), timing of biopsy (pre- vs post-hysterectomy) and type of biopsy (pipelle vs full-thickness). WHAT IS KNOWN ALREADY: Patient-derived biopsies are vital to endometrial research, but pre-analytical variables relating to their collection may affect downstream analysis, as is evident in other tissues. STUDY DESIGN SIZE DURATION: A prospective observational study including patients undergoing hysterectomy for endometrial cancer (EC) or benign indications was conducted at a large tertiary gynaecological unit in the UK. Endometrial biopsies were obtained at different time points (pre- or post-hysterectomy) using either a pipelle endometrial sampler or as a full-thickness wedge biopsy. PARTICIPANTS/MATERIALS SETTING METHODS: The changes in HIF1α, CA9, VEGFA and PR protein levels were measured by semi-quantitative analysis of immunostaining, and the expression levels of three genes (CA9, VEGFA and PR) were investigated by quantitative real-time PCR, in endometrial biopsies from 43 patients undergoing hysterectomy for EC (n = 22) or benign gynaecological indications (n = 21). MAIN RESULTS AND THE ROLE OF CHANCE: An increase in HIF1α immunostaining was observed in EC versus benign endometrium (functionalis glands) obtained pre-hysterectomy (P < 0.001). An increase in CA9 immunostaining was observed in EC versus benign endometrial functionalis glands at both pre- and post-hysterectomy time points (P = 0.03 and P = 0.003, respectively). Compared with benign endometrial pipelle samples, EC samples demonstrated increased mRNA expression of CA9 (pre-hysterectomy P < 0.001, post-hysterectomy P = 0.008) and VEGFA (pre-hysterectomy P = 0.004, post-hysterectomy P = 0.002). In benign uteri, HIF1α immunoscores (functionalis glands, P = 0.03 and stroma, P = 0.009), VEGFA immunoscores (functionalis glands, P = 0.03 and stroma, P = 0.01) and VEGFA mRNA levels (P = 0.008) were increased in matched post-hysterectomy versus pre-hysterectomy samples. Similarly, in EC, an increase in VEGFA immunoscores (epithelial and stromal) and VEGFA mRNA expression was observed in the matched post-hysterectomy versus pre-hysterectomy biopsies (P = 0.008, P = 0.004 and P = 0.018, respectively). Full-thickness benign post-hysterectomy endometrial biopsies displayed increased VEGFA (P = 0.011) and PR (P = 0.006) mRNA expression compared with time-matched pipelle biopsies. LARGE SCALE DATA: N/A. LIMITATIONS REASONS FOR CAUTION: This descriptive study explores the effect of pre-analytical variables on the expression of four proteins and three hypoxia-related genes in a limited number of endometrial biopsies from patients with EC and benign controls. Due to the small number, it was not possible to investigate other potential variables such as menstrual cycle phase, region-specific differences within the endometrium, grade and stage of cancer, and surgical technicalities. WIDER IMPLICATIONS OF THE FINDINGS: Careful consideration of the effects of these pre-analytical variables is essential when interpreting data relating to human endometrial biopsies. A standardized approach to endometrial tissue collection is essential to ensure accurate and clinically transferrable data. STUDY FUNDING/COMPETING INTERESTS: The authors have no conflicts of interest to declare. The work included in this manuscript was funded by Wellbeing of Women project grants RG1073 and RG2137 (D.K.H.), Wellbeing of Women Entry-Level Scholarship ELS706 and Medical Research Council MR/V007238/1 (A.M./D.K.H.), Liverpool Women's Hospital Cancer Charity (M.A.) and University of Liverpool (L.B., L.R. and E.N.).

Molecular cloning of the major surface antigen of leishmania.

The gene encoding gp63, the major surface glycoprotein of Leishmania promastigotes, was isolated from Leishmania major using a synthetic oligonucleotide probe based on the NH2-terminal protein sequence of purified gp63. DNA sequence analysis and the translated amino acid sequence indicate that gp63 is synthesized as precursor molecule having both an NH2-terminal preregion (signal peptide) and an adjacent proregion. This structure is consistent with the protease activity of gp63 since many other proteases are synthesized as precursor forms requiring processing for enzymatic activity. Hybridization studies demonstrated that there are multiple copies of the gp63 gene in the genome of L. major and other Leishmania species. The conservation of the coding sequence of gp63 amongst diverse species of Leishmania provides further support for the importance of gp63 during the life cycle of Leishmania.

Enzyme replacement therapy by fibroblast transplantation: long-term biochemical study in three cases of Hunter's syndrome.

We have assessed the effectiveness of transplanted histocompatible fibroblasts as a long-lived source of lysosomal enzymes for replacement therapy in three patients with Hunter's syndrome, over periods ranging from 2.5 to 3.75 yr. The level of Hunter corrective factor excreted by all three patients increased after transplantation, as did the activity of alpha-L-idurono-2-sulfate sulfatase in serum, when measured directly with a radioactive disulfated disaccharide substrate. Sulfatase activity was also raised in leukocyte homogenates from the two patients that we were able to assess. These increases in enzyme activity were accompanied by corresponding increases in catabolism of heparan and dermatan sulfates, as shown by (a) a decrease in sulfate:uronic ratios of urinary oligosaccharides, (b) an increase in iduronic acid monosaccharide, and (c) a normalization of Bio-Gel P-2 gel filtration profiles. Both the increase in enzyme activity and increased catabolism were maintained during the period of study and were not affected by either a gradual decrease or total withdrawal of immunosuppressive therapy.

The Saccharomyces cerevisiae chromosome III left telomere has a type X, but not a type Y', ARS region.

A yeast Saccharomyces cerevisiae telomeric region was isolated by chromosome walking from HML alpha, the most distal known gene on the chromosome III left (IIIL) end. The terminal heterodisperse 3.3-kilobase (kb) SalI fragment on chromosome IIIL, 8.6 kb distal to HML alpha, was cloned in a circular vector to generate a telomeric probe. Southern hybridization and DNA sequencing analyses indicated that 0.6 kb (+/- 200 base pairs) of 5'-C1-3A-3' simple tandem repeat sequence, adjacent to a 1.2-kb type X ARS region, constitutes the telomere on the chromosome IIIL end, and no type Y' ARS region homologies exist between HML alpha and the IIIL terminus.

Serotherapy of a patient with a monoclonal antibody directed against a human lymphoma-associated antigen.

A preliminary serotherapeutic trial was undertaken with a monoclonal antibody designated antibody 89 (Ab 89) directed against a lymphoma-associated antigen. In vitro studies demonstrated that Ab 89 could mediate complement-dependent lysis and macrophage adherence but not antibody-dependent cell-mediated cytotoxicity. To evaluate toxicity and therapeutic efficacy, two courses of Ab 89 were administered to a patient with an Ab 89-reactive tumor. Transient decreases in the number of circulating tumor cells and the appearance of circulating dead cells were noted with the infusion of Ab 89. Following administration of 150 mg or more of Ab 89, small amounts of antibody could be demonstrated on circulating tumor cells at a time when no free antibody was found in the serum. The inability to deliver a significant amount of Ab 89 to tumor cells in vivo is thought to be secondary to a circulating tumor antigen. Following each infusion, the amount of this blocking antigen decreased but could not be entirely cleared from the serum. This study provides preliminary evidence for the lack of clinical toxicity of a monoclonal antibody and identifies circulating blocking antigens as a significant obstacle to serotherapy.

Recombinant Leishmania surface glycoprotein GP63 is secreted in the baculovirus expression system as a latent metalloproteinase.

A gene encoding the Leishmania surface metalloproteinase, GP63, was modified using the polymerase chain reaction to obtain effective secretion of recombinant GP63 (reGP63) in the baculovirus insect cell expression system. The coding region for the N-terminal signal peptide (SP) of GP63 was modified to resemble the SP for the GP67 envelope protein from the budded virus form of Autographa californica nuclear polyhedrosis virus. To prevent processing at the C-terminus with a glycosyl phosphatidylinositol anchor and the subsequent membrane anchoring of reGP63 in insect cells, the coding region for a putative SP at the C-terminus of GP63 was deleted. The reGP63 protein was glycosylated and secreted as a latent metalloproteinase in the baculovirus expression system. The reGP63 protein was purified from serum-free medium using concanavalin A lectin affinity chromatography, with a yield of 1 mg/l. The purified Leishmania reGP63 was secreted as a latent proteinase. Treatment of reGP63 with HgCl2 resulted in activation of full proteinase activity and a concomitant decrease in M(r). The mechanism of the activation of Leishmania reGP63 is consistent with that of other members of the family of matrix-degrading metalloproteinases.

Selenium utilization in humans--a long-term, self-labeling experiment with stable isotopes.

A stable (nonradioactive) isotope of selenium in a chemical form common in foods (selenomethionine) or inorganic selenite was taken orally (200 micrograms/d) for 3 wk to label deep body pools. By deep body pools we mean selenium compartments that are large and/or have a slow turnover (exchange) rate. Blood plasma was removed, stored for 11 mo, and later reinfused as a labeled tracer dose with the selenium label in all of the biologically significant chemical forms. Accessible tissues such as red blood cells were highly labeled (20-25%) in the subjects receiving selenomethionine. Selenium from deep body pools is excreted primarily via the urine (80%). Reexcretion of previously absorbed selenium back into the gastrointestinal tract can be measured, avoiding a major source of error in conventional balance studies used to estimate nutrient absorption.

Heterogeneity of the genes encoding the major surface glycoprotein of Leishmania donovani.

The major surface glycoprotein of Leishmania (GP63) is present on all known species of Leishmania and likely plays an integral role during the infection of macrophages in the mammalian host. To identify regions of GP63 which may be of functional significance, the nucleotide sequence of a gene encoding GP63 of Leishmania donovani was determined and compared to the sequences reported for GP63 genes of Leishmania major and Leishmania chagasi. The GP63 nucleotide and predicted protein sequence was highly conserved among the 3 species despite their diverse geographical distribution. L. donovani GP63 is encoded by a multigene family and the gene locus contains at least 7 tandemly repeated genes and at least 3 genes which are dispersed from the tandem array. In addition, polymerase chain reaction and Southern blot analyses demonstrated that there was size heterogeneity within the pro-peptide coding regions of the multiple GP63 genes of L. donovani and that such genes were expressed concurrently in the promastigote life stage.

Modification of GP63 genes from diverse species of Leishmania for expression of recombinant protein at high levels in Escherichia coli.

Toward the future development of a defined subunit vaccine against leishmaniasis is, high levels of recombinant GP63 for diverse species of Leishmania were produced in Escherichia coli. Several features of Leishmania GP63 genes were simultaneously modified with the polymerase chain reaction (PCR) using either cloned genes or total genomic DNA from Leishmania as template DNA for the PCR amplification reactions. The PCR products included only the coding region for the predicted mature form of GP63 that occurs on the surface of Leishmania, flanked by the appropriate translation signals and cloning sites for the production of recombinant GP63 as nonfusion protein in E. coli. When the codon usage in the GP63 gene was modified to reduce the guanine and cytosine content for the codons adjacent to the ATG initiation codon, rGP63 represented about 50% of total protein in E. coli. Mouse monoclonal antibodies raised against purified Leishmania major rGP63 had equivalent immunoblotting characteristics for native GP63 and recombinant GP63 with respect to linear determinants on GP63 expressed in diverse species of Leishmania. Human T cell lines and clones were derived from a patient infected with Leishmania braziliensis panamensis using rGP63 purified from an L. major GP63 expression clone as antigen.

The major surface glycoprotein (GP63) is present in both life stages of Leishmania.

Leishmania exist as extracellular promastigotes which multiply in the gut of the sandfly insect vector and as intracellular amastigotes which divide in the phagolysosome of mononuclear phagocytic cells of the mammalian host. Promastigotes express a major surface glycoprotein of 63 kDa, referred to as GP63. The expression of GP63 in both Leishmania life stages was studied using rabbit antibodies against native GP63 as well as rabbit antibodies against recombinant GP63 that was synthesized in an Escherichia coli expression system. Immunofluorescence staining detected GP63 in intracellular amastigotes contained within a macrophage cell line and within freshly isolated lesion amastigotes. Western blot analysis using anti-recombinant GP63 antibodies also demonstrated that amastigotes synthesize GP63 which may undergo differential post-translational processing as compared to promastigote GP63.

Genes encoding the major surface glycoprotein in Leishmania are tandemly linked at a single chromosomal locus and are constitutively transcribed.

The major surface glycoprotein of Leishmania, gp63, is encoded by a small multi-gene family of tandemly linked genes which map to a single chromosome. For Leishmania major, there are five 3.1 kilobasepair (kb) direct repeat units which include a 1.8-kb open reading frame and a 1.3-kb intergenic or spacer region. In addition, there is a single gene copy linked as a direct repeat but separated from the tandem array of gp63 genes by about 8 kb. The restriction enzyme map of the repeat unit is highly conserved among the gene copies. The regions which flank the tandemly repeated genes diverge outside of the 3.1-kb repeat unit. Transcription of the gp63 gene locus is constitutive as the 3-kb transcript was present in promastigotes as well as in amastigotes. A minor 6-kb gp63 gene transcript was also detected in Northern blot analysis which could signify the transcription of the genes as a polycistronic or multigene precursor RNA.

Blood volume changes in patients with cystic fibrosis.

Simultaneous red blood cell (RBC) and plasma volume determinations were obtained in 16 patients with cystic fibrosis (CF) and moderately severe pulmonary involvement. Hypervolemia with an increase in both RBC and plasma volumes was observed. Changes in blood volume were marked when values were indexed by weight but less significant when indexed by height. Decreasing systemic arterial oxygen saturation was associated with a progressive increase in RBC mass, hematocrit value, and hemoglobin level and a decrease in mean corpuscular hemoglobin concentration. RBC and total blood volumes were highest in patients with cor pulmonale and congestive heart failure. However, the compensatory polycythemic response in patients with CF was inadequate when compared with the response to hypoxemia in patients with cyanotic congenital heart disease. The insufficient oxygen-carrying capacity may compromise tissue oxygen delivery and necessitate treatment.

Use of homologous platelet survival in differential diagnoses of chronic thrombocytopenia in childhood.

Twenty-one children with chronic thrombocytopenia of obscure etiology, 19 of whom had normal numbers of bone marrow megakaryocytes, were evaluated in order to determine if factors extrinsic to the platelets themselves (presumably antibodies) caused the thrombocytopenia. The rapid destruction of platelets by such antibodies has been demonstrated to cause idiopathic thrombocytopenic purpura (ITP). History, physical examination, and routine laboratory studies were unable to provide an exact diagnosis in these patients. Measurement of platelet survival, using sodium chromate Cr 51 labeled homologous platelets obtained from unrelated donors, permitted a clear separation of these children into two groups. One group (11 patients) had a markedly shortened platelet life span, and was considered to have chronic ITP. The other group (10 patients) had a normal or only slightly reduced platelet life span. Thus, the destruction of platelets by extrinsic factors could be ruled out. Since the diagnosis of chronic ITP was excluded, splenectomy could be avoided in these patients since it is rarely useful in children with nonimmune forms of chronic thrombocytopenia. Homologous platelet survival measurement is a useful diagnostic tool in certain children with chronic thrombocytopenia.

Genetic heterogeneity in Peruvian Leishmania isolates.

Leishmania were isolated from Peruvian patients with uta or espundia; genomic DNA was examined for restriction fragment length differences by Southern blot analysis using DNA probes for beta-tubulin and for the major surface antigen gp63. Using 5 different restriction endonucleases, Peruvian isolates show homogeneity when examined at the beta-tubulin locus. In contrast, the organisms demonstrated heterogeneity both within and between disease groups when examined for restriction site differences within the gp63 locus. The differences observed did not correlate with the 2 disease groups. Comparison of these Peruvian isolates to New World reference strains of the Leishmania braziliensis complex reveals no consistent pattern of identity with either L. b. guyanensis, L. b. panamensis, or L. b. braziliensis.

Screening of the equine intestinal microflora for potential probiotic organisms.

REASONS FOR PERFORMING STUDY: Probiotics have not been demonstrated to provide any beneficial health effects in horses, possibly because of improper selection of probiotic organisms. This study was designed to identify lactic acid bacteria of equine origin with predetermined beneficial properties which might make them useful as therapeutic probiotics. HYPOTHESIS: A small percentage of lactic acid bacteria that are native to the intestinal tract of horses possess properties that may be useful in the treatment and/or prevention of gastrointestinal disease in horses. METHODS: Faecal samples were collected from healthy mature horses and foals. Lactic acid bacteria were isolated and tested for the ability to grow in acid and bile environments, aerotolerance and in vitro inhibition of enteropathogens. One isolate that possessed these properties was administered orally to healthy mature horses and foals and gastrointestinal survival was assessed. RESULTS: Of the 47 tested organisms, 18 were deemed to be adequately acid- and bile-tolerant. All were aerotolerant. Four organisms markedly inhibited Salmonella spp. One isolate, Lactobacillus pentosus WE7, was subjectively superior and chosen for further study. It was also inhibitory against E. coli, moderately inhibitory against S. zooepidemicus and C. difficile and mildly inhibitory against C. perfringens. After oral administration, this isolate was recovered from the faeces of 8/9 (89%) foals and 7/8 (87.5%) mature horses. CONCLUSIONS: Lactobacillus pentosus WE7 possesses in vitro and in vivo properties that may be useful for the prevention and treatment of enteric disease in horses. POTENTIAL RELEVANCE: The beneficial in vitro and in vivo properties that L. pentosus WE7 possesses indicate that randomised, blinded, placebo-controlled efficacy studies are warranted.

Anna's story: narratives of personal experience about being labeled learning disabled.

The narratives of personal experience of an adolescent named Anna provided insights into two issues: first, how well her discourse conformed to linguistic expectations for the types of narrative traditionally deemed acceptable in school, and second, the themes associated with the presentation of self that Anna and her peers addressed when talking about their personal experiences as students labeled learning disabled. By narrative, we mean the root metaphor for human sense-making that is described in the following epigraph by Bruner. We found that Anna's narrative differed from typical school-based expectations in that its structure was reminiscent of the oral tradition. From the group of students, we heard themes of isolation, undervaluing, and oppression. We recommend a more thoughtful and respectful approach to educational decision making that gives voice to students.

Hospitalized incidence and case fatality for upper gastrointestinal bleeding from 1999 to 2007: a record linkage study.

BACKGROUND: Upper gastrointestinal (GI) bleeding is the most common emergency managed by gastroenterologists. AIM: To establish the hospitalized incidence and case fatality for upper GI bleeding, and to determine how they are associated with factors including day of admission, hospital size, social deprivation and distance from hospital. METHODS: Systematic record linkage of hospital in-patient and mortality data for 24 421 admissions for upper GI bleeding among 22 299 people in Wales from 1999 to 2007. RESULTS: The hospitalized incidence of upper GI bleeding was 134 per 100 000. Case fatality was 10.0%. Incidence was stable from 1999 to 2007; case fatality fell from 11.4% in 1999-2000 to 8.6% in 2006-7. Incidence was associated significantly with social deprivation. Compared with weekday admissions, case fatality was 13% higher for weekend admissions and 41% higher for admissions on public holidays. There was little variation in case fatality according to social deprivation, hospital size or distance from hospital. CONCLUSIONS: Incidence, but not case fatality, was associated significantly with social deprivation. The higher mortality for weekend and public holiday admissions could not be explained by measures of case mix and may indicate a possible impact of reduced staffing levels and delays to endoscopy at weekends in some hospitals.