Expression patterns of ciliopathy genes ARL3 and CEP120 reveal roles in multisystem development.

BACKGROUND: Joubert syndrome and related disorders (JSRD) and Jeune syndrome are multisystem ciliopathy disorders with overlapping phenotypes. There are a growing number of genetic causes for these rare syndromes, including the recently described genes ARL3 and CEP120. METHODS: We sought to explore the developmental expression patterns of ARL3 and CEP120 in humans to gain additional understanding of these genetic conditions. We used an RNA in situ detection technique called RNAscope to characterise ARL3 and CEP120 expression patterns in human embryos and foetuses in collaboration with the MRC-Wellcome Trust Human Developmental Biology Resource. RESULTS: Both ARL3 and CEP120 are expressed in early human brain development, including the cerebellum and in the developing retina and kidney, consistent with the clinical phenotypes seen with pathogenic variants in these genes. CONCLUSIONS: This study provides insights into the potential pathogenesis of JSRD by uncovering the spatial expression of two JSRD-causative genes during normal human development.

The United Kingdom Primary Immune Deficiency (UKPID) Registry: report of the first 4 years' activity 2008-2012.

This report summarizes the establishment of the first national online registry of primary immune deficency in the United Kingdom, the United Kingdom Primary Immunodeficiency (UKPID Registry). This UKPID Registry is based on the European Society for Immune Deficiency (ESID) registry platform, hosted on servers at the Royal Free site of University College, London. It is accessible to users through the website of the United Kingdom Primary Immunodeficiency Network (www.ukpin.org.uk). Twenty-seven centres in the United Kingdom are actively contributing data, with an additional nine centres completing their ethical and governance approvals to participate. This indicates that 36 of 38 (95%) of recognized centres in the United Kingdom have engaged with this project. To date, 2229 patients have been enrolled, with a notable increasing rate of recruitment in the past 12 months. Data are presented on the range of diagnoses recorded, estimated minimum disease prevalence, geographical distribution of patients across the United Kingdom, age at presentation, diagnostic delay, treatment modalities used and evidence of their monitoring and effectiveness.

Embryonic and foetal expression patterns of the ciliopathy gene CEP164.

Nephronophthisis-related ciliopathies (NPHP-RC) are a group of inherited genetic disorders that share a defect in the formation, maintenance or functioning of the primary cilium complex, causing progressive cystic kidney disease and other clinical manifestations. Mutations in centrosomal protein 164 kDa (CEP164), also known as NPHP15, have been identified as a cause of NPHP-RC. Here we have utilised the MRC-Wellcome Trust Human Developmental Biology Resource (HDBR) to perform immunohistochemistry studies on human embryonic and foetal tissues to determine the expression patterns of CEP164 during development. Notably expression is widespread, yet defined, in multiple organs including the kidney, retina and cerebellum. Murine studies demonstrated an almost identical Cep164 expression pattern. Taken together, these data support a conserved role for CEP164 throughout the development of numerous organs, which, we suggest, accounts for the multi-system disease phenotype of CEP164-mediated NPHP-RC.

SonoAVC: a novel method of automatic volume calculation.

OBJECTIVES: To assess the ability of the new software SonoAVC to measure follicular volume and to compare these volume calculations with those made by conventional methods. METHODS: Three-dimensional ultrasound imaging was used to acquire volumetric data from the ovaries of 51 women undergoing controlled ovarian stimulation as part of in-vitro fertilization treatment. All assessments were performed on the day of oocyte retrieval and the true volume of each follicle was ascertained by manual measurement of the follicular aspirate. SonoAVC was used to automatically measure the volume of follicles and to provide three perpendicular diameters (xyz diameters), which were used to estimate volume using the sphere formula. The sphere formula was also used to estimate the volume from manual measurements of follicle diameter derived from conventional two-dimensional (2D) displays. Virtual Organ Computer-aided AnaLysis (VOCAL) was also used to measure volume, and the validity of each technique was compared using limits of agreement. RESULTS: Two hundred and twenty-four follicles with a mean follicular volume of 3.7 (range, 0.4-16.2) cm(3) were studied. SonoAVC provided highly accurate automatic follicular volume measurements in all cases. Volume estimations made from the automatic maximal follicular diameter measurements (xyz diameters) were less valid. VOCAL proved highly valid but was less accurate than SonoAVC. Volumes estimated from manually derived follicular diameter measurements were the least accurate. CONCLUSIONS: SonoAVC provides highly valid, automatic measurements of follicular volume. These measurements are more accurate than volumes estimated from 2D manual measurements, automated measurements of follicular diameter and those calculated using VOCAL.

Measurement of the longitudinal energy distribution of electrons in low energy beams using electrostatic elements.

The Transverse Energy Spread Spectrometer (TESS) was designed primarily to study the mean transverse energy spread of electrons emitted from photocathode electron sources at both room and liquid nitrogen temperatures as a function of quantum efficiency through analysis of the photoemission footprint. By reconfiguring the potentials applied to different detector elements, TESS can also be used to measure the mean longitudinal energy spread of photoemitted electrons. Initial plans were to use electrostatic wire meshes as a retarding element which prevents the detection of electrons with insufficient energy to overcome a variable potential barrier. However, this method has proved impractical and a new method has been proposed in which the photocathode bias potential is swept (effectively from a state of no electron emission to full emission) and the emitted photocurrent is then detected by using a photoemitted charge collector. In this article, we present the TESS set-up and analyze this new method to measure the longitudinal energy distribution curve. Experimental results are presented and compared to simulated results by utilising a custom designed tracking code.

Analysis of the factors that influence the Finnegan Neonatal Abstinence Scoring System.

OBJECTIVE: To evaluate factors that can influence the Finnegan Neonatal Abstinence Score (FNAS). STUDY DESIGN: Retrospective analysis of 367 patients admitted to two level IV neonatal intensive care units. Linear mixed effects models were developed to evaluate daily census, time of the day, and day of the week as fixed effect predictors. The degree of influence that nurses had on FNAS variability was also estimated. RESULTS: Bivariate analyses showed that daily census and the time of day have significant influence on the FNAS in institution 1, with minimal clinical significance. The proportion of variation in the FNAS attributable to differences in nurses was of 9.8% and 5.1% for institutions 1 and 2, respectively (P<0.0001). CONCLUSIONS: The minimal influences of extraneous factors on the FNAS support the clinical utility of the scoring system in the assessment and management of infants with Neonatal Abstinence Score.

Experimental metastasis and differentiation of murine melanoma cells: actions and interactions of factors affecting different intracellular signalling pathways.

Various factors that modulate the differentiation of malignant cells are known to affect their experimental metastatic potential (EMP), or lung colonization after intravenous injection into syngeneic animals. However, some results and conclusions on the relation between cell differentiation and metastasis have appeared to conflict. We have reanalysed this by measurement of EMP of B16 melanoma sublines after culture with agents or conditions that acted on differentiation through various intracellular pathways. All tested agents did affect the EMP. EMP was usually positively correlated with differentiation under diverse conditions, but exceptions showed that there is no direct causal connection. Nor could all findings be explained in terms of cell proliferation or expression of major histocompatibility antigens. Some data helped to explain disparities between previous reports. Specific novel findings included the following. The stimulation of EMP by melanocyte-stimulating hormone (MSH) as well as all other tested effects of MSH were prevented by extended exposure to 12-O-tetradecanoyl phorbol acetate (TPA), suggesting a requirement for protein kinase C activity as well as G-protein coupling in MSH action. Cells grown with cholera toxin were always more differentiated than untreated cells, but the EMP could be either markedly increased or markedly decreased by cholera toxin under different conditions. The basic culture medium apparently determined this striking reversal. The EMP was also significantly affected by the extracellular pH.

Recurrent posterior thigh symptoms detrimental to performance in rugby union: predisposing factors.

Recurrent hamstring injury is a very common problem in rugby union, but has been largely ignored in the literature. It is concluded that a multifactorial aetiology may be present, and that these symptoms may be part of a continuum of symptoms that may lead to more serious injuries. Effective management needs to focus not just locally, but include proximal issues such as lumbo-pelvic stability and correcting lumbar spine dysfunction. The factors addressed in a prevention programme are likely to contribute to performance gains.

Genetic counseling for the deaf.

Genetic counseling is a process that emphasizes accurate diagnosis of hereditary conditions and communication of information to families. Genetic counseling involves systematic collection of family and medical history, a physical examination by a certified clinical geneticist, sharing of information with the family, and follow-up and support services. The issues that arise in genetic counseling can differ for every family and are often dependent on the degree of deafness present in the family, age of onset, and linguistic and cultural orientation. It is important for the genetic counselor to consider these factors in the provision of genetic services. With the increasing application of molecular genetics to the diagnosis and management of hereditary deafness and the increasing participation of families with deafness in research studies, the involvement of genetic counselors to provide information and education to consumers as well as medical professionals and researchers is becoming even more critical. The success of genetic counseling for the provision of information to families and the delineation of types of hereditary deafness through clinical and laboratory research is dependent on appropriate referrals by medical professionals, including otolaryngologists. A working relationship between otolaryngologists and clinical geneticists for the referral and evaluation of patients with hereditary deafness or deafness of "unknown" etiology is important.

Moderate levels of hypohydration impairs bowling accuracy but not bowling velocity in skilled cricket players.

The effects of exercise-induced hypohydration on the motor skill performance of cricket bowling was examined in seven medium-fast bowlers who performed a random order of two experimental trials. Trials consisted of a bowling test (36 deliveries; PREBOWL) in a thermoneutral (16+/-2 degrees C) environment followed by approximately 1 hr of intermittent exercise in a heated environment (28+/-2 degrees C) and a further thermoneutral bowling test (36 deliveries; POSTBOWL). During one trial fluid intake was restricted (HYPO) whereas in the other, subjects were forced to drink in an effort to maintain euhydration (EUH). During all bowling tests subjects were provided with a fixed target on a cricket pitch and the line, length, and velocity of each delivery was determined. Pre-trial hydration status was confirmed by similar body mass (BM; 89.5+/-13.7 vs. 88.9+/-13.4 kg) and haemoglobin concentration (15.0+/-0.8 vs. 14.8+/-0.8 g.100 ml(-1) for EUH and HYPO, respectively). BM loss was greater in HYPO than EUH (2.48+/-0.58 vs. 0.46+/-0.45 kg). Accordingly, the resultant hypohydration was higher after HYPO than EUH (2.78+/-0.49 vs. 0.47+/-0.41% of BM). Whereas HYPO had no effect on bowling velocity (102+/-4 vs. 105+/-8 km x h(-1)), univariate analyses revealed independent differences for both bowling line (2.9+/-0.5 vs 3.4+/-0.6, P<0.01) and length (2.9+/-0.5 vs 3.4+/-0.6, P<0.01) of delivery after HYPO. We conclude that moderate (-2.8% of BM) exercise-induced hypohydration has minimal effect on maximal bowling velocity, but there is a detrimental effect on skilled motor performance in well-trained subjects.

Use of magnifying lens to aid neonatal umbilical arterial catheter insertion.

We conducted this trial to test the hypothesis that use of a magnifying lens would facilitate umbilical arterial catheter (UA C) insertion in neonates. Neonates <33 weeks' gestation requiring a UA C were randomized to conventional method of catheter insertion or an experimental method using a 2.0 X magnifying lens mounted on a headband. A total of 44 neonates (Conventional = 23, Experimental = 21) with comparable demographic characteristics completed the study. The time taken for (primary outcome) and rate of successful UAC insertion (secondary outcome) were not significantly different between the two groups (median times: conventional: 88 seconds,experimental: 70 seconds, P = 0. 734) (Success rate: conventional: 19/23 (83%) vs experimental:13/21 (63%), P = 0. 1791 Thus, our hypothesis was rejected.

Accuracy of the clinical diagnosis of Down syndrome.

OBJECTIVES: To determine the accuracy of clinical diagnosis of Down syndrome, identify problems in reaching a diagnosis, to provide recommendations for improvement and estimate a minimum prevalence for all types of Down syndrome. DESIGN: A retrospective observational study was carried out over a five-year period. Genesis, a database located in the Department of Medical genetics, was used to identify the number of Down syndrome karyotypes including trisomy, translocation, and mosaic sample variants. Age of diagnosis was determined using date of receipt. Karyotyping requests for a clinical diagnosis of Down syndrome were also identified. Patient notes and cytogenetic laboratory reports were used to identify clinical indication for karyotyping. SETTING: Regional Genetics Centre, covering all cytogenetic analyses for referrals within the entire Northern Ireland population. RESULTS: 208 postnatal cases of Down syndrome were identified, 197 (94.7%) trisomy, 3 (1.45%) translocation, and 8 (3.85%) mosaic variants. 112 (54.8%) were male and 96 (46.2%) female. 268 samples were taken to confirm or exclude a clinical diagnosis of Down syndrome. 185 of these had Down syndrome, 77 were normal, and 6 had another abnormality. 90% and 100% of trisomy and translocation Down syndrome respectively were diagnosed on the basis of clinical features. This fell to 37.5% of mosaic Down syndrome patients being diagnosed clinically (p < 0.001). Simian crease, sandal gap, epicanthic folds, hypotonia, upslanting palpebral fissures, and protruding tongue are the most frequent characteristic features seen. Similarly epicanthic folds, protruding tongue, simian crease and sandal gap, hypotonia, and upslanting palpebral fissures are also described in a significant proportion of karyotypically normal individuals, thus arousing a suspicion of Down syndrome. 89.4% of patients were diagnosed between day 1 and 7 of life. Of 10.6% patients diagnosed after day 7 of life, 7.6% were adults and 3% children. The minimum prevalence was estimated at 167.9 per 100,000, or 1 in 595 births. CONCLUSION: In a defined population, with a prevalence of around 1 in 600 births, accurate clinical diagnosis occurred in 90%, 100%, and 37.5% of trisomy, translocation, and mosaic patients. 49.5% of patients had one or more of the following phenotypic findings: Simian crease, sandal gap, epicanthic folds, hypotonia, upslanting palpebral fissures, and protruding tongue. However, the same six features aroused a suspicion of Down syndrome in individuals with normal karyotyping, thus causing undue stress and worry to parents. Mosaic cases may be more common than previously recognised, and often do not have dysmorphic features. It is therefore a diagnosis that should always be considered in those who are educationally subnormal without a definitive diagnosis.

Use of baclofen for withdrawal in a preterm infant.

Baclofen is a gamma-aminobutyric acid agonist used primarily as a muscle relaxant to treat spasticity in children and adults. Withdrawal of oral baclofen is known to cause a withdrawal syndrome in adults. Only one previous case describes a withdrawal syndrome in a term infant, manifested by seizures, associated with the use of oral baclofen in the mother. This case describes a withdrawal syndrome and the unique use of baclofen for withdrawal in a preterm infant.

Natural killer cell cytotoxicity in patients with recurrent herpes infections: diagnostic utility of a flow cytometric assay.

BACKGROUND: Primary immune deficiencies of natural killer (NK) cells have been described in patients with a susceptibility to herpes infections. AIMS: To assess the diagnostic utility of measurement of NK cytotoxicity in patients with recurrent oral herpes infections. METHODS: A retrospective audit was carried out on results obtained over an 18-month period, from 28 NK cell cytotoxicity assays (24 patients; all with a history of recurrent oral herpes infections), and 24 control samples (three healthy donors). Percentage specific cytotoxicity (PSC) was determined by measurement of the percentage of K562 target cells lysed by NK cells after incubation, using the NK TEST. Comparison of PSC was made with reference ranges provided. RESULTS: No patient with absent NK/NKT cells or NK cell cytotoxicity was identified (95% CI 0 to 14.8%). Two patients had persistently low PSC. Two patients with reduced PSC showed PSC within the normal reference range on repeat testing. Patient and control samples were seen both above and below the reference ranges. A relationship was expected between NK cell percentage and PSC; however this correlation was not significant (r(s)=0.29, p=0.18, 95% CI -0.14 to 0.63). CONCLUSIONS: A deficiency of NK cell cytotoxicity has not been identified in this cohort. An apparent reduction in cytotoxicity may be due to normal interpersonal and intersample variability in NK cytotoxicity. Without reference ranges established from a large population of control samples to account for this, a reduction in PSC is difficult to define. Further studies are required to identify if a correlation exists between the percentage of NK cells and PSC.

Germline mutation prevalence in the base excision repair gene, MYH, in patients with endometrial cancer.

Germline mutations in the base excision repair gene, MutY human homolog (MYH), have recently been associated with a recessively inherited multiple adenoma polyposis syndrome and colorectal cancer. The spectrum of extracolonic lesions is still being characterized, although preliminary reports suggest that bi-allelic mutation carriers may share some of the clinical features of other hereditary colon cancer syndromes. Of 225 endometrial cancer patients, we identified one individual as a compound heterozygote, carrying mutations Y165C and G382D of MYH, and five individuals with heterozygous defects (three G382D and two Y165C). The patient with the bi-allelic Y165C/G382D mutation also had a sebaceous carcinoma, a feature of Muir-Torre syndrome. Although several intronic polymorphisms were detected in the heterozygous carriers, no other pathogenic variants were identified. While not conclusive, this novel and interesting finding provides evidence that bi-allelic germline mutations in MYH may increase susceptibility to endometrial cancer.

The comparative assessment of paraspinal tissue compliance in asymptomatic female and male subjects in both prone and standing positions.

Segmental bilateral paraspinal tissue compliance measures were obtained from 50 male and 50 female asymptomatic subjects (vertebral segments C7-L5). Interexaminer concordance for the taking of the measure was found to be extremely high (r greater than .90). Additionally, test-retest measures obtained from the same subjects initially, and again 15 min or 2 wk later indicated fairly low short-term as well as long-term temporal variabilities for the measure. The greatest levels of variability were encountered at lumbar segments, particularly with 2 wk intervals between assessments, and when measures were obtained with subjects in the standing, rather than prone, position. Data generated by this study suggest that tissue compliance assessments may provide useful information with respect to the contractile state of the paraspinal musculature at various regions or segmental levels.

Batch process solar disinfection is an efficient means of disinfecting drinking water contaminated with Shigella dysenteriae type I.

AIMS: The mortality and morbidity rate caused by Shigella dysenteriae type I infection is increasing in the developing world each year. In this paper, the possibility of using batch process solar disinfection (SODIS) as an effective means of disinfecting drinking water contaminated with Sh. dysenteriae type I is investigated. METHODS: Phosphate-buffered saline contaminated with Sh. dysenteriae type I was exposed to simulated solar conditions and the inactivation kinetics of this organism was compared with that of Sh. flexneri, Vibrio cholerae and Salmonella typhimurium. SIGNIFICANCE: Recovery of injured Sh. dysenteriae type I may be improved by plating on medium supplemented with catalase or pyruvate. Sh. dysenteriae type I is very sensitive to batch process SODIS and is easily inactivated even during overcast conditions. Batch process SODIS is an appropriate intervention for use in developing countries during Sh. dysenteriae type I epidemics.