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Nuclear pore complexes (NPCs) regulate nucleocytoplasmic transport and are anchored in the nuclear envelope by the transmembrane nucleoporin NDC1. NDC1 is essential for post-mitotic NPC assembly and the recruitment of ALADIN to the nuclear envelope. While no human disorder has been associated to one of the three transmembrane nucleoporins, biallelic variants in AAAS, encoding ALADIN, cause triple A syndrome (Allgrove syndrome). Triple A syndrome, characterized by alacrima, achalasia, and adrenal insufficiency, often includes progressive demyelinating polyneuropathy and other neurological complaints. In this report, diagnostic exome and/or RNA sequencing was performed in seven individuals from four unrelated consanguineous families with AAAS-negative triple A syndrome. Molecular and clinical studies followed to elucidate the pathogenic mechanism. The affected individuals presented with intellectual disability, motor impairment, severe demyelinating with secondary axonal polyneuropathy, alacrima, and achalasia. None of the affected individuals has adrenal insufficiency. All individuals presented with biallelic NDC1 in-frame deletions or missense variants that affect amino acids and protein domains required for ALADIN binding. No other significant variants associated with the phenotypic features were reported. Skin fibroblasts derived from affected individuals show decreased recruitment of ALADIN to the NE and decreased post-mitotic NPC insertion, confirming pathogenicity of the variants. Taken together, our results implicate biallelic NDC1 variants in the pathogenesis of polyneuropathy and a triple A-like disorder without adrenal insufficiency, by interfering with physiological NDC1 functions, including the recruitment of ALADIN to the NPC., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Increasingly long and complex informed consents have yielded studies demonstrating comparatively low participant comprehension and satisfaction with traditional face-to-face approaches. In parallel, interest in electronic consents for clinical and research genomics has steadily increased, yet limited data are available for trio-based genomic discovery studies. We describe the design, development, implementation, and validation of an electronic iConsent application for trio-based genomic research deployed to support genomic studies of cerebral palsy. iConsent development incorporated stakeholder perspectives including researchers, patient advocates, institutional review board members, and genomic data-sharing considerations. The iConsent platform integrated principles derived from prior electronic consenting research and elements of multimedia learning theory. Participant comprehension was assessed in an interactive teachback format. The iConsent application achieved nine of ten proposed desiderata for effective patient-focused electronic consenting for genomic research. Overall, participants demonstrated high comprehension and retention of key human subjects' considerations. Enrollees reported high levels of satisfaction with the iConsent, and we found that participant comprehension , iConsent clarity , privacy protections , and study goal explanations were associated with overall satisfaction . Although opportunities exist to optimize iConsent, we show that such an approach is feasible, can satisfy multiple stakeholder requirements, and can realize high participant satisfaction and comprehension while increasing study reach., Competing Interests: Conflicts of Interest Paul Gross is President and CEO of the CP Research Network, which contributed to the funding of this project. Mr. Gross personally made financial contributions (donations) to CPRN to support this work, but receives no financial compensation related to either. Dr. Noritz has consulted for Abbott Nutrition, unrelated to this project. Dr. Shrader receives research funding from NIH and serves on the National Advisory Board for Medical Rehabilitative Research for NIH/NICHD.

Highly conserved transport protein particle (TRAPP) complexes regulate subcellular trafficking pathways. Accurate protein trafficking has been increasingly recognized to be critically important for normal development, particularly in the nervous system. Variants in most TRAPP complex subunits have been found to lead to neurodevelopmental disorders with diverse but overlapping phenotypes. We expand on limited prior reports on TRAPPC6B with detailed clinical and neuroradiologic assessments, and studies on mechanisms of disease, and new types of variants. We describe 29 additional patients from 18 independent families with biallelic variants in TRAPPC6B. We identified seven homozygous nonsense (n = 12 patients) and eight canonical splice-site variants (n = 17 patients). In addition, we identified one patient with compound heterozygous splice-site/missense variants with a milder phenotype and one patient with homozygous missense variants. Patients displayed non-progressive microcephaly, global developmental delay/intellectual disability, epilepsy and absent expressive language. Movement disorders including stereotypies, spasticity and dystonia were also observed. Brain imaging revealed reductions in cortex, cerebellum and corpus callosum size with frequent white matter hyperintensity. Volumetric measurements indicated globally diminished volume rather than specific regional losses. We identified a reduced rate of trafficking into the Golgi apparatus and Golgi fragmentation in patient-derived fibroblasts that was rescued by wild-type TRAPPC6B. Molecular studies revealed a weakened interaction between mutant TRAPPC6B (c.454C>T, p.Q152*) and its TRAPP binding partner TRAPPC3. Patient-derived fibroblasts from the TRAPPC6B (c.454C>T, p.Q152*) variant displayed reduced levels of TRAPPC6B as well as other TRAPP II complex-specific members (TRAPPC9 and TRAPPC10). Interestingly, the levels of the TRAPPC6B homologue TRAPPC6A were found to be elevated. Moreover, co-immunoprecipitation experiments showed that TRAPPC6A co-precipitates equally with TRAPP II and TRAPP III, while TRAPPC6B co-precipitates significantly more with TRAPP II, suggesting enrichment of the protein in the TRAPP II complex. This implies that variants in TRAPPC6B may preferentially affect TRAPP II functions compared to TRAPP III functions. Finally, we assessed phenotypes in a Drosophila TRAPPC6B-deficiency model. Neuronal TRAPPC6B knockdown impaired locomotion and led to wing posture defects, supporting a role for TRAPPC6B in neuromotor function. Our findings confirm the association of damaging biallelic TRAPPC6B variants with microcephaly, intellectual disability, language impairments, and epilepsy. A subset of patients also exhibited dystonia and/or spasticity with impaired ambulation. These features overlap with disorders arising from pathogenic variants in other TRAPP subunits, particularly components of the TRAPP II complex. These findings suggest that TRAPPC6B is essential for brain development and function, and TRAPP II complex activity may be particularly relevant for mediating this function., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Guarantors of Brain. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Colorectal cancer (CRC) is more prevalent in south-central Asian countries, particularly the Afghan population. Screening for CRC in the Afghan population has always been challenging, primarily due to the tribal and social cultures, lack of facilities, and lack of education. The United States (US) will soon face a significantly massive influx of Afghan refugees. It becomes imperative to initiate and implement effective measures regarding CRC screening in these refugee populations. The current review article aims to identify the most likely challenges faced for CRC screening in this Afghan refugee population in the US and address the possible measures to overcome these challenges., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2022, Waheed et al.)

Spermatogenesis-associated 5 like 1 (SPATA5L1) represents an orphan gene encoding a protein of unknown function. We report 28 bi-allelic variants in SPATA5L1 associated with sensorineural hearing loss in 47 individuals from 28 (26 unrelated) families. In addition, 25/47 affected individuals (53%) presented with microcephaly, developmental delay/intellectual disability, cerebral palsy, and/or epilepsy. Modeling indicated damaging effect of variants on the protein, largely via destabilizing effects on protein domains. Brain imaging revealed diminished cerebral volume, thin corpus callosum, and periventricular leukomalacia, and quantitative volumetry demonstrated significantly diminished white matter volumes in several individuals. Immunofluorescent imaging in rat hippocampal neurons revealed localization of Spata5l1 in neuronal and glial cell nuclei and more prominent expression in neurons. In the rodent inner ear, Spata5l1 is expressed in the neurosensory hair cells and inner ear supporting cells. Transcriptomic analysis performed with fibroblasts from affected individuals was able to distinguish affected from controls by principal components. Analysis of differentially expressed genes and networks suggested a role for SPATA5L1 in cell surface adhesion receptor function, intracellular focal adhesions, and DNA replication and mitosis. Collectively, our results indicate that bi-allelic SPATA5L1 variants lead to a human disease characterized by sensorineural hearing loss (SNHL) with or without a nonprogressive mixed neurodevelopmental phenotype., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021. Published by Elsevier Inc.)

In addition to commonly associated environmental factors, genomic factors may cause cerebral palsy. We performed whole-exome sequencing of 250 parent-offspring trios, and observed enrichment of damaging de novo mutations in cerebral palsy cases. Eight genes had multiple damaging de novo mutations; of these, two (TUBA1A and CTNNB1) met genome-wide significance. We identified two novel monogenic etiologies, FBXO31 and RHOB, and showed that the RHOB mutation enhances active-state Rho effector binding while the FBXO31 mutation diminishes cyclin D levels. Candidate cerebral palsy risk genes overlapped with neurodevelopmental disorder genes. Network analyses identified enrichment of Rho GTPase, extracellular matrix, focal adhesion and cytoskeleton pathways. Cerebral palsy risk genes in enriched pathways were shown to regulate neuromotor function in a Drosophila reverse genetics screen. We estimate that 14% of cases could be attributed to an excess of damaging de novo or recessive variants. These findings provide evidence for genetically mediated dysregulation of early neuronal connectivity in cerebral palsy.

CLN6-Batten disease is a rare neurodegenerative disorder with no cure, characterized by accumulation of lipofuscin in the lysosome, glial activation, and neuronal death. Here we test the therapeutic efficacy of modulating collapsin response mediator protein 2 (CRMP2) activity via S -N-benzy-2-acetamido-3-methoxypropionamide (( S )-Lacosamide) in a mouse model of CLN6-Batten disease. Promisingly, mouse neuronal cultures as well as Cln6 patient fibroblasts treated with varying concentrations of ( S )-Lacosamide showed positive restoration of lysosomal associated deficits. However, while acute in vivo treatment enhanced glial activation in 3-month-old Cln6 mutant mice, chronic treatment over several months did not improve behavioral or long-term survival outcomes. Therefore, modulation of CRMP2 activity via ( S )-Lacosamide alone is unlikely to be a viable therapeutic target for CLN6-Batten disease., Competing Interests: The authors declare that there are no competing interests associated with the manuscript., (© 2019 The Author(s).)

Question I recently evaluated a child in my clinic after an emergency department visit where she presented having woken up that morning refusing to walk and was crawling around the house. The parents reported she was getting over a cold, and I recall similar cases of myositis during the H1N1 influenza epidemic a few years ago. What are the key features of myositis that I should recognize? Which investigations are needed to confirm the diagnosis and how should affected patients be managed? Answer Benign acute childhood myositis is a mild and self-limited sudden onset of lower extremity pain during or following recovery from a viral illness. Presentation can include tiptoe gait or refusal to walk, secondary to symmetric bilateral lower extremity pain that resolves quickly, usually within 3 days. In general, no investigation is needed except in severe cases for which screening bloodwork and a urine myoglobin test can confirm the diagnosis and rule out complications. Myoglobinuria and highly elevated creatine phosphokinase levels are rare but should be a consideration for admission to hospital. Prognosis is excellent and management might include rest and analgesia., (Copyright© the College of Family Physicians of Canada.)

Neuronal ceroid lipofuscinoses (NCLs; also known collectively as Batten Disease) are a family of autosomal recessive lysosomal storage disorders. Mutations in as many as 13 genes give rise to ∼10 variants of NCL, all with overlapping clinical symptomatology including visual impairment, motor and cognitive dysfunction, seizures, and premature death. Mutations in CLN6 result in both a variant late infantile onset neuronal ceroid lipofuscinosis (vLINCL) as well as an adult-onset form of the disease called Type A Kufs. CLN6 is a non-glycosylated membrane protein of unknown function localized to the endoplasmic reticulum (ER). In this study, we perform a detailed characterization of a naturally occurring Cln6 mutant (Cln6(nclf)) mouse line to validate its utility for translational research. We demonstrate that this Cln6(nclf) mutation leads to deficits in motor coordination, vision, memory, and learning. Pathologically, we demonstrate loss of neurons within specific subregions and lamina of the cortex that correlate to behavioral phenotypes. As in other NCL models, this model displays selective loss of GABAergic interneuron sub-populations in the cortex and the hippocampus with profound, early-onset glial activation. Finally, we demonstrate a novel deficit in memory and learning, including a dramatic reduction in dendritic spine density in the cerebral cortex, which suggests a reduction in synaptic strength following disruption in CLN6. Together, these findings highlight the behavioral and pathological similarities between the Cln6(nclf) mouse model and human NCL patients, validating this model as a reliable format for screening potential therapeutics.

Aims: Accurate determination of HER-2 status is critical to identify patients for whom trastuzumab treatment will be of benefit. Although the recommended primary method of evaluation is immunohistochemistry, numerous reports of variability in interpretation have raised uncertainty about the reliability of results. Recent guidelines have suggested that image analysis could be an effective tool for achieving consistent interpretation, and this study aimed to assess whether this technology has potential as a diagnostic support tool., Methods and Results: Across a cohort of 275 cases, image analysis could accurately classify HER-2 status, with 91% agreement between computer-aided classification and the pathology review. Assessment of the continuity of membranous immunoreactivity in addition to intensity of reactivity was critical to distinguish between negative and equivocal cases and enabled image analysis to report a lower referral rate of cases for confirmatory fluorescence in situ hybridization (FISH) testing. An excellent concordance rate of 95% was observed between FISH and the automated review across 136 informative cases., Conclusions: This study has validated that image analysis can robustly and accurately evaluate HER-2 status in immunohistochemically stained tissue. Based on these findings, image analysis has great potential as a diagnostic support tool for pathologists and biomedical scientists, and may significantly improve the standardization of HER-2 testing by providing a quantitative reference method for interpretation.

For over 30 years, the African cichlid fish, Astatotilapia burtoni, has been an important model system for studying the mechanisms underlying socially mediated behavioral change, with the focus being the dominance behavior of males. A recently collected wild-stock (WS) of this species invigorates interest in parallel studies of females' behavior. Here, we describe a robust 'good-mother' phenotype, increased maternal affiliation in fry, and subtle differences in males' behavior that are exhibited by this new stock. While the females of both the laboratory-stock (LS) and the WS brood the developing fry in their buccal cavity, only the WS continues to provide maternal care after initial release of the fry while the LS engage in filial cannibalism. We show that weight loss during starvation, either during brooding or with restriction of food, is greater in the LS than in the WS; thus, the observed behavioral differences may be tied to metabolic differences. The WS also exhibits a robust androgen response to challenge during the maternal care phase. Given the increasing power of genomic tools available for this species, the comparison of these two stocks will offer the opportunity to investigate the genetic and genomic basis of behavioral differences.

Aims: A retrospective post-mortem analysis was undertaken to determine the aetiology of sudden cardiac death (SCD) amongst individuals aged

Objectives: Evidence about sociodemographic factors associated with late attendance for antenatal care in the UK is of poor quality. This study aimed to identify any social or ethnic differences in access to antenatal care, and to quantify the effect of any such differences using data collected in a survey of women's experiences of antenatal screening., Study Design: Cross-sectional survey using a postal questionnaire., Methods: A stratified clustered random sampling strategy was used. Hospitals in England were stratified according to ethnic mix. In order to ensure inclusion of an adequate number of women from Black and Minority Ethnic (BME) backgrounds, hospitals with >or= 15% of women of BME origin were oversampled. Pregnant women aged >or= 16 years, receiving care in 15 participating hospitals, were sent a postal questionnaire at 27-31 weeks of gestation. Logistic regression was used to estimate odds ratios (ORs) comparing social and ethnic groups for attendance for antenatal care, adjusting for sociodemographic and clinical factors., Results: In total, 839 women (57%) returned completed questionnaires. Compared with all women giving birth in 2005 in England and Wales, the survey sample contained fewer women aged <20 years (5.8% vs 6.9%), more women aged >35 years (24.1% vs 19.6%) and fewer women who were born outside the UK (14.8% vs 20.8%). Five percent of responders were late attenders for their first antenatal appointment. The odds of late initiation of antenatal care were higher for women born outside the UK [OR 4.37, 95% confidence interval (CI) 2.25-8.52; P=0.0004] and for women living without a husband/partner (OR 2.74, 95% CI 1.81-4.16; P=0.0002). In total, 2.5% of women were late attenders for their booking appointment. The odds of late booking were higher for Black women (OR 5.92, 95% CI 2.97-11.83) and women living without a husband/partner (OR 1.95, 95% CI 0.97-3.93; P=0.06)., Conclusions: A small proportion of women initiate and/or book late for antenatal care. This study provides recent, good-quality evidence that women born outside the UK and those living without a husband/partner may be at particular risk of late attendance for antenatal care.

Background and Aims: Trastuzumab provides clinical benefit for advanced and early breast cancer patients whose tumours over-express or have gene amplification of the HER2 oncogene. The UK National External Quality Assessment Scheme (NEQAS) for immunohistochemical testing was established to assess and improve the quality of HER2 immunohistochemical testing. However, until recently, no provision was available for HER2 fluorescence in situ hybridisation (FISH) testing. A pilot scheme was set up to review the performance of FISH testing in clinical diagnostic laboratories., Methods: FISH was performed in 6 reference and 31 participating laboratories using a cell line panel with known HER2 status., Results: Using results from reference laboratories as a criterion for acceptable performance, 60% of all results returned by participants were appropriate and 78% either appropriate or acceptable. However, 22.4% of results returned were deemed inappropriate, including 13 cases (4.2%) where a misdiagnosis would have been made had these been clinical specimens., Conclusions: The results of three consecutive runs show that both reference laboratories and a proportion of routine clinical diagnostic (about 25%) centres can consistently achieve acceptable quality control of HER2 testing. Data from a significant proportion of participating laboratories show that further steps are required, including those taken via review of performance under schemes such as NEQAS, to improve quality of HER2 testing by FISH in the "real world".

Background: The majority of cases of breast cancer scoring HER2 weak positive (2+) on immunohistochemistry (IHC) using the HercepTest are not associated with amplification of the HER2/neu gene., Aim: To examine the reproducibility of IHC in cases scoring 2+ subsequently shown to have gene amplification by fluorescence in-situ hybridisation (FISH)., Methods: A retrospective analysis of 153 cases referred for FISH confirmation of a weak positive HercepTest (2+) result was performed. Repeat IHC was undertaken in cases with weak positive (2+) referral IHC and amplification of the HER2 gene by FISH., Results: Amplification of the HER2 gene was confirmed in 29/153 cases (19%) scoring 2+ on IHC. Repeat IHC was carried out on 25 IHC 2+ cases: 7 (28%) scored 2+ on repeat IHC, 18 (72%) scored 3+ and were reclassified as strong positive. A heterogeneous expression pattern was present in 3/17 cases scoring 3+., Conclusions: The majority of HercepTest 2+ results are not accompanied by gene amplification and represent "false positive" IHC in terms of prognostic or therapeutic relevance. A small proportion of HercepTest 2+ scores represent true 2+ IHC positive cases accompanied by gene amplification: a category probably biologically related to 3+ IHC cases. The remainder of cases of HercepTest 2+ accompanied by gene amplification represent a category of referral IHC 2+ weak positive, FISH amplified, repeat IHC 3+ strong positive, best described as "false negative 2+ IHC". This has implications for selection of cases for FISH analysis where weak positive (2+) IHC score is used as a triage for FISH testing, and for testing strategies in referral laboratories undertaking FISH analysis.

HER2 and TOP2A genes, located on 17q, can be coamplified in cancer. Overexpression of both genes has been reported in high-grade, androgen-resistant prostate cancer. Both genes have not been compared in a single prostate cancer study and the frequency of TOP2A amplifications in prostate cancer is unknown. Using tissue microarrays, we did immunohistochemistry and fluorescence in situ hybridization for HER2 and TOP2A in 100 prostate cancers (41 localized and 59 advanced) and 42 cases of benign prostatic hyperplasia (BPH). Amplification was defined as a target/centromere signal ratio of > or =1.5. HER2 immunohistochemistry was scored from 0 to 3+. Percentage nuclei staining for topoisomerase IIalpha (topoIIalpha) was recorded; overexpression was defined as > or =5% cells staining. Eighteen (31%) advanced prostate cancers showed topoIIalpha overexpression; 12 (26%) showed TOP2A low-level amplification; 9 (16%) expressed HER2; and 6 (13%) showed HER2 low-level amplification. No high-level amplification of either gene (target/centromere signal ratio of > or =3.0) was detected. TOP2A coexpression and coamplification were seen in 75% and 66% of HER2-positive cases, respectively. Localized prostate cancer or BPH showed no gene amplification or topoIIalpha overexpression. Gene amplification or overexpression correlated with high stage and Gleason score. The presence of TOP2A amplification in advanced cancer was associated with androgen resistance and decreased survival by multivariate analysis. This is the first study to document low-level TOP2A amplification in prostate cancer and an association with reduced survival. TOP2A amplification may occur with or without HER2 duplication and is often associated with topoIIalpha expression. Therapies directed against topoIIalpha (and HER2) in such patients may improve survival.

Objective: To develop a questionnaire to assess patients' views of clinical trials, and to report the results from the questionnaire in two patient groups: asthma and cancer., Design: A 43 item questionnaire asking patients about their views to clinical trials was developed on the basis of interviews with trialists and focus groups with patients. The questionnaire was mailed to patients with a diagnosis of either asthma or cancer. A set of items was then selected, via statistical analyses, to form the core of the questionnaire., Participants: Patients with a diagnosis of cancer in one NHS Hospital Trust, and patients with a diagnosis of asthma in two NHS Hospital Trusts., Results: Completed questionnaires were received from 353 cancer patients and 578 asthma patients. Factor analyses of the data indicated that 22 items contributed to five dimensions: 'positive beliefs', 'safety', 'information needs', 'negative expectations' and 'patient involvement'. Differences between asthma and cancer patients on these dimensions were small. A regression of these dimension scores against a variable asking if patients would be willing to take part in trials found that 'safety' and 'information needs' did not contribute significantly to the model for either asthma or cancer patients., Conclusions: A questionnaire has been developed for use in assessing patients' views towards clinical trials. Results from the surveys reported here suggest that patient views about the importance of trials and beliefs about the value of patient involvement are likely to be predictive of whether or not patients will agree to take part in a study.

Patients on certain waiting lists in the UK National Health Service (NHS) are now offered the choice of persevering with their home hospital or switching to another hospital where they will be treated on a guaranteed date. Such decisions require knowledge of performance. We used facilitated focus groups to investigate the views of patients and members of the public on publication of information about the performance of healthcare providers. Six groups with a total of 50 participants met in six different locations in England. Participants felt that independent monitoring of healthcare performance is necessary, but they were ambivalent about the value of performance indicators and hospital rankings. They tended to distrust government information and preferred the presentational style of 'Dr Foster', a commercial information provider, because it gave more detailed locally relevant information. Many participants felt the NHS did not offer much scope for choice of provider. If public access to performance information is to succeed in informing referral decisions and raising quality standards, the public and general practitioners will need education on how to interpret and use the data.

The U.S. military currently faces serious difficulties in training medical personnel in peacetime for the tasks of war. The military beneficiary population comprises fit young service men and women, their dependents, and retirees. Their peacetime care, although vital, does little to prepare military medical personnel for war. Medical commanders have instituted an array of training programs to compensate for this shortfall, but there remains a large gap between operational medical needs and training opportunities in peacetime. The military has begun to examine whether simulation can fill this gap. An array of commercial, off-the-shelf technologies are already being used with varying degrees of success, and major initiatives are under way in both academia and industry, supported by the military, to develop virtual reality products for combat medical training. Even as the military exploits emerging technology and begins to articulate a simulation strategy, there is a growing interest in civilian medicine in the potential for simulation to affect patient safety--how medical simulation might mitigate the injuries and deaths caused by medical errors--and how it might also improve the quality of medical education and training.

We examined a panel of 21 patients diagnosed with compulsive buying for two DNA sequence polymorphisms found in the gene that encodes the serotonin transport (5-HTT). One polymorphism, found in the promoter region of the 5-HTT gene, involves a 44-base pair (bp) deletion, and the other, found in the second intron, is due to variable numbers of a repeat sequence. We also typed a panel of 38 psychiatrically normal controls for both 5-HH markers. When compared to this control panel, no significant differences were seen for either 5-HTT marker among the compulsive buyers.

For a variety of technical reasons it is rarely possible to study cytogenetic abnormalities in ductal carcinoma in situ (DCIS) using traditional techniques. However, by combining molecular biology and computerized image analysis it is possible to carry out cytogenetic analyses on formalin-fixed, paraffin-embedded tissue, using comparative genomic hybridization (CGH). The purpose of this study was to identify the prevalence of chromosomal amplifications and deletions in high-grade DCIS and to look specifically for unique or consistent abnormalities in this pre-invasive cancer. Twenty-three cases of asymptomatic, non-palpable, screen-detected, high-grade DCIS were examined using CGH on tumour cells obtained from histology slides. All cases showed chromosomal abnormalities. A wide variety of amplifications and deletions were spread across the genome. The most frequent changes were gains of chromosomes 17 (13 of 23), 16p (13 of 23), and 20q (9 of 23) and amplifications of 11q13 (22 of 23), 12q 24.1-24.2 (12 of 23), 6p21.3 (11 of 23), and 1q31-qter (6 of 23). The most frequent deletions were on 13q 21.3-q33 (7 of 23), 9p21 (4 of 23), and 6q16.1 (4 of 23). These findings indicate that high-grade DCIS is, from a cytogenetic viewpoint, an advanced lesion. There was no absolutely consistent finding in every case, but amplification of 11q13 was found in 22 of the 23 cases. The precise significance of this is unknown at present. This region of chromosome 11q harbours a number of known oncogenes, including cyclin D1 andINT2. It is likely that many of these findings are the result of accumulated chromosomal abnormalities, reflecting an unstable genome in established malignancy., (Copyright 1999 John Wiley & Sons, Ltd.)

The serine hydroxymethyltransferase (SHMT) gene family is composed of three distinct loci. The cytosolic (cSHMT) and mitochondrial (mSHMT) genes constitute the functional members of the gene family, while the third member, SHMT-ps1, is a processed pseudogene descended from cSHMT. PCR analysis of 38 primate and nonprimate mammal species indicates that the reverse transcription event that gave rise to SHMT-ps1 might have occurred after the divergence of the primates from the rest of the mammals. In addition, direct sequencing of primate PCR products has revealed several features--including two deletions, an insertion, and two single base mutations--that are unique to specific phylogenetic branches of the order Primates. These unique features make the SHMT-ps1 locus a useful marker in molecular studies of the primates.

The D3-dopamine receptor gene, DRD3, has been considered as a candidate gene in several disorders in which the dopaminergic system has been implicated including Tourette syndrome and schizophrenia. The DRD3 studies to date have all used as the gene marker a Bal I polymerase chain reaction restriction fragment length polymorphism (PCR RFLP). There have been recent reports on a second marker, an Msp I PCR RFLP, that lies 40 kb downstream. We have typed a sample of 16 Tourette syndrome families with both markers and observed significant linkage disequilibrium between the two markers but no apparent association of either marker with Tourette syndrome.

Aneuploidy is an important prognostic factor in many cancers. Chromosome 1 abnormalities are present in most breast carcinomas. These may be part of a non-specific increase in DNA (aneuploid status) or represent a restricted chromosomal abnormality. In 16 breast carcinomas we compared chromosome 1 aneusomy with ploidy status. Patients were selected from a mammographically screened population and interphase tumour nuclei were studied by in situ hybridization using a chromosome 1 pericentromeric probe. Ploidy status was assessed by image cytometry on disaggregated cells from paraffin blocks. Of eight cases showing chromosome 1 aneusomy, six (75%) were aneuploid and two diploid. Six (75%) of the eight eusomic cases were aneuploid. This study demonstrates that chromosome 1 aneusomy does not always reflect a gross aneuploid status but, in some tumours, is part of a more restricted chromosomal abnormality. Interphase cytogenetics, possibly using a small panel of pericentromeric probes, may be more sensitive than DNA cytometry for detecting abnormal nuclear DNA content.

Estimation of oestrogen receptors (ER) in breast carcinoma has important therapeutic and prognostic implications. The dextran-coated charcoal (DCC) biochemical technique is being replaced in many institutions by immunohistochemical techniques. The aim of this study was to assess the level of agreement between ER counts in frozen and paraffin sections. Ninety-seven infiltrating breast carcinomas were studied immunohistochemically, for ER in frozen and paraffin sections. Cases were classified as ER-positive if > or = 10% of the tumor cells were positive. Both techniques gave a high level of correlation and agreement. There was complete agreement in classifying cases as ER-positive or ER-negative. ER estimation on only one case was outside 2SD of complete agreement. Oestrogen-receptor content can be assayed satisfactorily in paraffin sections and offers many advantages over the frozen section technique.

Aggressive behavior is difficult to capture with most available rating instruments because it occurs with low frequency. The Overt Aggression Scale (OAS) was designed to capture all incidents on a 24-hour basis. To assess its usefulness, the OAS was piloted on 16 hospitalized, severely aggressive children with conduct disorder. OAS ratings capture episodes of aggressive behavior, measure trends in aggressive behavior over time, and appear to reflect change associated with pharmacotherapy in hospitalized children. This study suggests that the use of the OAS on a children's psychiatric inpatient unit is appropriate and feasible for clinical as well as research purposes.

The present study reports on the frequency of MDM2 gene amplification and MDM2 protein expression in a series of 100 breast carcinomas and its association with accumulation of the p53 protein. Of the 100 cases, frozen samples for 82 cases were available for Southern blotting. Three of the 82 (4%) demonstrated MDM2 gene amplification of up to 6-fold. Immunohistochemical analysis of the formalin-fixed, paraffin-embedded tumours demonstrated that 7/97 (7%) had nuclear expression for MDM2 in 10-50% of the tumour cells (type 2 staining) and were denoted MDM2+. Two of the MDM2-amplified samples were MDM2+ with one of the two tumours also displaying type 2 p53 nuclear staining. Finally at the protein level, MDM2+ tumours were significantly associated with tumours having low levels of p53 staining (0-10% cells positive) (P = 0.03). We conclude that MDM2 gene amplification occurs at a lower frequency in breast cancer than in non-epithelial tumours. Alterations in MDM2 and p53 may represent alternative pathways in tumorigenesis, but they are not mutually exclusive in all cases.

Because of the relative rarity of ductal carcinoma in situ (DCIS) in the premammographic screening era, the unavailability of adequate fresh tissue for culture, and the lack of cytogenetic expertise in most pathology departments, there is little information on karyotypic abnormalities in DCIS. The purpose of this study was to investigate the frequency of chromosome 1 aneusomy in DCIS, using interphase cytogenetic techniques, and to correlate the findings with nuclear grade. Twenty-one cases of DCIS, identified in a mammographically screened population, were studied by in situ hybridization. Chromosome 1 numbers were identified in interphase nuclei in conventional histology sections, using a specific centromeric probe (pUC 1.77). In each case, 100 tumour nuclei were compared with 100 normal nuclei. Eighteen of 21 (86 per cent) cases were aneusomic for chromosome 2. This included 15 of 16 (94 per cent) pure comedo or predominantly comedo DCIS. Fifteen of 16 (94 per cent) DCIS with grade 3 nuclei and 3 of 5 (60 per cent) cases with grade 2 nuclei were aneusomic. One case with grade 3 nuclei (a comedo carcinoma) was negative. We conclude that chromosome 1 aneusomy precedes invasion and is a relatively consistent occurrence in those DCIS with high nuclear grade.

Aims: To examine the pattern of reactivity of Ki-67 in neuroblastoma and correlate this with a) clinical prognostic criteria and b) cell cycle statistics (using flow cytometry)., Methods: Four micron sections of paraffin embedded (PE) tissue from 55 patients (25 pre chemotherapy and 30 post) were placed on to aminosialinised slides, dewaxed and rehydrated. Slides were pretreated in a microwave oven, endogenous peroxidase activity blocked using 3% hydrogen peroxide and Ki-67 reactivity investigated using a streptavidin/biotin/peroxidase technique. DNA ploidy was also performed from an immediately adjacent section on the same block using a FACScan and Cellfit software., Results: Ki-67 reactivity was well defined and highly reproducible. Eighteen out of 30 post chemotherapy samples were totally negative, despite evidence of proliferation on flow cytometry and all subsequently died of disease. As interpretation post chemotherapy was therefore deemed unreliable, this group was excluded from analysis. Reactivity in pretreatment samples ranged from 0% to 67%; staining was restricted to the nucleus with a distinct pattern noted in the nucleolus. Ki-67 positivity was lower in aneuploid compared with diploid tumours (mean 26% vs 36%, NS). Among diploid tumours, a lower percentage positivity was noted in those patients with better clinical prognostic parameters. Correlation however between Ki-67 and SG2M phases of cell cycle was poor (RS = 0.39, NS)., Conclusion: Assessment of proliferation in neuroblastoma by Ki-67 reactivity in paraffin embedded tissue is reliable in pretreatment samples and can be incorporated into routine immunohistochemical evaluation. Larger multicentre studies are required to further evaluate Ki-67 reactivity as a prognostic indicator.

Benign and malignant breast lesions in cytologic preparations can be difficult to distinguish. The authors applied the silver nucleolar organizer region (AgNOR) technique to cytologic preparations obtained from surgical specimens to evaluate its diagnostic usefulness in making this distinction. Sixty-two benign and 36 malignant lesions were examined. AgNORs were counted and evaluated using a subjective scoring technique to examine AgNOR size, shape, and clustering. The benign lesions had a mean count of 4.44 AgNORs/nucleus (95% CI, 2.4-6.5) and the malignant cases, 9.52 AgNORs/nucleus (95% confidence interval, 7.4-11.7; P < .0005). The median score for benign cases was 7 and for malignant cases, 13 (P < .0001). With a few exceptions, cases with high counts had high scores. The diagnostic accuracy of combined counting and pattern assessment was 90%. The likelihood ratio for correct diagnosis using AgNOR counting was 14:1 and for AgNOR scoring, 13:1.

Pigmented solar keratosis (PSK) is sometimes clinically indistinguishable from lentigo maligna, a form of malignant melanoma in situ. Occasionally histologic diagnosis is also difficult. Accurate diagnosis is essential, as the treatment and prognosis for each condition differs considerably. To determine whether there was a significant overlap in the number of melanocytes in these sun-damaged skin lesions, or whether immunohistochemistry might be helpful in the differential diagnosis, the authors examined skin biopsy specimens from 26 patients with obvious lentigo maligna and 15 patients with PSK using 3 monoclonal antibodies (HMB-45, NK1C3, and vimentin) and 1 polyclonal antibody (S-100 protein). Formalin-fixed paraffin sections were immunostained with each of the above antibodies, and immunopositive cells per mm2 of epidermis were counted. The difference between lentigo maligna and PSK counts was statistically significant at a level of P < .0001; furthermore, there was almost no overlap between the two groups. The sensitivity for the diagnosis of lentigo maligna was high with all antibodies. However, HMB-45 had the highest sensitivity and the lowest false-positive rate and was visually most pleasing. Using a cut-off count of 60 cells per mm2 of epidermis, HMB-45 had a sensitivity of 96% and a 0% false-positive rate. In this study, lentigo maligna was easily differentiated from PSK. The real value of immunohistochemistry in the differential diagnosis of these pigmented lesions should be tested in a prospective study using cases that are difficult to diagnose by routine light microscopy.

ABSTRACT Two new cases are reported of persistent tardive dyskinesia associated with neuroleptic treatment of patients with Tourette's disorder. Previously, 44 cases were described in 8 published reports, including 36 children and adolescents, but diagnostic criteria were infrequently specified. In our review of these cases, using the criteria of Schooler and Kane but modified by Gualtieri's more conservative duration criteria of 16 weeks, only 2 of these cases were consistent with a diagnosis of persistent tardive dyskinesia. The 2 new cases are reported here. A 12-year-old, who was treated with haloperidol 4 mg daily since the age of 8, developed fine vermicular movements of the tongue of moderate severity. Despite discontinuation of the neuroleptic, symptoms of tardive dyskinesia still persisted at age 15 and were associated with difficulty in speech production. A 42-year-old, who was treated with haloperidol 1 mg three times daily for 7 years, developed jaw movements and lip smacking that persisted for more than 2 years. Abnormal involuntary movement scale (AIMS) ratings supported a diagnosis of tardive dyskinesia in both patients with Tourette's disorder. The identification of tardive dyskinesia in the setting of a preexisting movement disorder is discussed. Features that helped distinguish the movements of tardive dyskinesia and Tourette's disorder in these patients included the premonitory urges of Tourette's symptoms and a differential response of the symptoms to distracting voluntary motor tasks. Clinicians should be attentive and thorough in searching for symptoms of tardive dyskinesia following treatment with relatively low doses of haloperidol in patients with Tourette's disorder.

Cell proliferation can be studied by a variety of techniques. However, most require fresh tissue. To evaluate cell proliferation in formalin-fixed paraffin-embedded sections, the authors immunohistochemically studied 35 tumors and 11 samples of normal/hyperplastic tissue with PC10, a monoclonal antibody directed against proliferating cell nuclear antigen. Results were compared with those obtained with Ki-67 on fresh tissues. There was no significant difference between proliferating cell nuclear antigen and Ki-67 counts, which were strongly correlated (r = 0.8). Proliferating cell nuclear antigen positivity was easier to evaluate because morphology was better preserved in formalin-fixed tissue. The authors conclude that PC10 is an alternative to Ki-67 in evaluating cell proliferation and has the advantage of reacting with formalin-fixed paraffin-embedded tissue.

This open pilot study explored the efficacy and safety of pimozide, over a 3-week period, in hospitalized autistic children. Eight males, ages 4.2 to 8.3 years, completed the study. Intellectual functioning ranged from moderate to profound mental retardation. Symptoms included severe withdrawal, stereotypies, hyperactivity and/or hypoactivity, aggressiveness, and temper tantrums. Therapeutic daily doses of pimozide ranged from 3.0 mg to 6.0 mg with a mean of 4.9 mg (0.12-0.32 mg/kg; mean, 0.22). Laboratory studies including electrocardiogram and liver function tests remained within normal limits. Untoward effects were minimal and transient. Decreases of behavioral symptoms were evidenced on all measures including the Children's Psychiatric Rating Scale, Clinical Global Impressions, and Global Clinical Judgments Scale (consensus rating). Of the 5 hypoactive children, 4 showed a decrease in hypoactivity, whereas 1 child showed worsening. These findings are promising and indicate the need for further study.

The effects of methylphenidate on hospitalized conduct-disordered (CD) adolescents were examined by using teacher ratings of behavior, a measure of classroom learning, and a test of impulsivity. Twenty-two male adolescents with CD, 12-18 years of age, participated in a double-blind, placebo-controlled, within-subject (crossover) design in which each adolescent received three doses of methylphenidate (10 mg, 15 mg, and 20 mg) and a placebo in a randomly assigned, counterbalanced order. Seven of the adolescents had a comorbid diagnosis of attention deficit hyperactivity disorder. Significant overall medication effects were shown on teacher ratings of conduct, and on number of arithmetic questions correctly completed and time spent. Within the limitations of this study, stimulant actions may be effective for some aspects of CD in the absence of attention deficit hyperactivity disorder, although only for specific measures.

A review of arterial embolism to upper and lower limbs from 1961 to 1985. During this period 253 patients were treated for 269 episodes of embolus at the Princess Alexandra Hospital. The period was divided into 5 year intervals and analysis showed significantly increasing age for the population over this period. Similarly a decreasing mortality rate was noted during this period although limb salvage did not change after the initial 5 year period. There was a rising incidence of atherosclerotic ischaemic heart disease and a decreasing incidence of rheumatic heart disease. The overall mortality rate was 15.6% while the overall amputation rate was 8.2%. Factors significantly related to survival were acute myocardial infarction and level of occlusion. Factors significantly relating to morbidity were pre-operative heparin administration and performance of completion angiography. The performance of completion angiography was shown to improve the result from embolectomy without having adverse influence on the mortality rate. The results of this review indicate that preoperative heparinization, the performance of completion angiography, and the flexibility to explore multiple levels of the limb circulation, are essential factors in the management of peripheral arterial embolism.

The successful management of 2 patients is reported, each involving the closure of a fistula into the inferior vena cava as a sequel to the rupture of an atherosclerotic aortic aneurysm. It is an uncommon complication which should be suspected in the presence of an aortic aneurysm with a loud machinery bruit. Associated peripheral cyanosis and venous pulsation in the accessible proximal veins of the lower extremity and priapism afford strong supporting evidence.

One hundred and sixty-eight ruptured aortic aneurysms presenting at the Princess Alexandra Hospital in an 11-year period have been reviewed. Surgical treatment still carries a high mortality and a very high postoperative morbidity. The survival rate in the Vascular Unit of this hospital has improved during the period studied and in the last five years has risen to 61%. The operative technique is described in detail, and the operative problems and complications are discussed. It is felt that prompt surgical treatment, and accurate assessment of blood loss with adequate replacement during surgery, combined with attention to early postoperative problems, particularly those of a respiratory nature, are the most important factors in survival.

A retrospective analysis of 75 patients with 103 atherosclerotic popliteal aneurysms, seen at Princess Alexandra Hospital, Brisbane, was undertaken to assess the results of management and the most appropriate approach to the contralateral popliteal aneurysm. These patients were seen between January 1968 and December 1985. There were no females, the mean age was 66.3 years and 36% of the patients had bilateral popliteal aneurysms. There was a high incidence of other associated aneurysms, abdominal aortic (21), femoral (12) and isolated iliac (3). Thrombosis in a popliteal aneurysm was the most common reason for presentation initially (48%), followed by detection of a lump in the popliteal fossa (22.6%), embolic phenomena (10.6%) and ruptured popliteal aneurysm (four cases). The most common presentation for contralateral aneurysms was detection of a lump in the popliteal fossa. Management of the contralateral popliteal aneurysm was classified as early, elective repair (n = 14) and no surgery or surgery delayed until the development of symptoms (n = 14). Outcome was classified as successful, retained and fully functional limb at follow-up; or failure, amputation or debilitating ischaemia. There was one failure among those undergoing early elective repair and seven failures among those undergoing no or delayed surgery (P = 0.033, Fisher's Exact Test). There were 79 reconstructions. Bypass and ligation (n = 54) resulted in two amputations and two long-term occlusions, interposition augmented vein graft (n = 15) in two amputations and no long-term occlusions and interposition Dacron grafts (n = 10) with no amputations and three long-term occlusions.(ABSTRACT TRUNCATED AT 250 WORDS)

Rupture of a Dacron arterial prosthesis is a most unusual complication, although it has been reported previously. Three further such cases are reported with a brief review of the relevant literature.

Actual HL-A typing has been performed on 115 members of cancer family N, a large kindred (over 1000 members ascertained) showing the findings consistent with the cancer family syndrome. In the cancer-prone line (branches C and D) of the family, 20 of 21 members with cancer had one HL-A haplotype, HL-A2-HL-A12 (relative odds = 6.30), including some decreased family members who had haplotypes assigned. Eleven of 12 family members with cancer in branches C and D, actually typed, had HL-A2-HL-A12 (relative odds = 6.06). The single exception showing cancer and another haplotype in branch D is a child of a family member with haplotype HL-A2-HL-A12.

Carcinoma of the colon was studied in Omaha-Douglas County, Nebraska (population 345,000). A total of 154 cases of colon cancer were diagnosed in 1964 (44.7/100,000). The frequency distribution of these patients in specific census tracts of this community was determined. Statistical analysis of the data showed a greater frequency of colon cancer in patients living in census tracts with higher average income. Colon cancer appears to be nonrandomly distributed with respect to the income and socioeconomic status of its victims, suggesting that hypotheses consistent with environmental variables--particularly those characterizing extremely high versus extremely low socioeconomic groups, including occupation, diet and other life patterns--should be pursued. All of these data have implications for cancer epidemiology, cancer control, and carcinogenesis.