RESUMO
BACKGROUND: Prune belly syndrome (PBS) is a rare, multi-system congenital myopathy primarily affecting males that is poorly described genetically. Phenotypically, its morbidity spans from mild to lethal, however, all isolated PBS cases manifest three cardinal pathological features: 1) wrinkled flaccid ventral abdominal wall with skeletal muscle deficiency, 2) urinary tract dilation with poorly contractile smooth muscle, and 3) intra-abdominal undescended testes. Despite evidence for a genetic basis, previously reported PBS autosomal candidate genes only account for one consanguineous family and single cases. METHODS: We performed whole exome sequencing (WES) of two maternal adult half-brothers with syndromic PBS (PBS + Otopalatodigital spectrum disorder [OPDSD]) and two unrelated sporadic individuals with isolated PBS and further functionally validated the identified mutations. RESULTS: We identified three unreported hemizygous missense point mutations in the X-chromosome gene Filamin A (FLNA) (c.4952 C > T (p.A1448V), c.6727C > T (p.C2160R), c.5966 G > A (p.G2236E)) in two related cases and two unrelated sporadic individuals. Two of the three PBS mutations map to the highly regulatory, stretch-sensing Ig19-21 region of FLNA and enhance binding to intracellular tails of the transmembrane receptor ß-integrin 1 (ITGß1). CONCLUSIONS: FLNA is a regulatory actin-crosslinking protein that functions in smooth muscle cells as a mechanosensing molecular scaffold, transmitting force signals from the actin-myosin motor units and cytoskeleton via binding partners to the extracellular matrix. This is the first evidence for an X-linked cause of PBS in multiple unrelated individuals and expands the phenotypic spectrum associated with FLNA in males surviving even into adulthood.
Assuntos
Filaminas/genética , Genes Ligados ao Cromossomo X/genética , Doenças Genéticas Ligadas ao Cromossomo X/genética , Síndrome do Abdome em Ameixa Seca/genética , Adulto , Doenças Genéticas Ligadas ao Cromossomo X/fisiopatologia , Predisposição Genética para Doença , Genótipo , Hemizigoto , Humanos , Masculino , Pessoa de Meia-Idade , Mutação de Sentido Incorreto/genética , Linhagem , Fenótipo , Síndrome do Abdome em Ameixa Seca/fisiopatologia , Sequenciamento do ExomaRESUMO
OBJECTIVE: To design a novel system of scoring prune belly syndrome (PBS) phenotypic severity at any presenting age and apply it to a large pilot cohort. PATIENTS AND METHODS: From 2000 to 2017, patients with PBS were recruited to our prospective PBS study and medical records were cross-sectionally analysed, generating individualised RUBACE scores. We designed the pragmatic RUBACE-scoring system based on six sub-scores (R: renal, U: ureter, B: bladder/outlet, A: abdominal wall, C: cryptorchidism, E: extra-genitourinary, generating the acronym RUBACE), yielding a potential summed score of 0-31. The 'E' score was used to segregate syndromic PBS and PBS-plus variants. The cohort was scored per classic Woodard criteria and RUBACE scores compared to Woodard category. RESULTS: In all, 48 males and two females had a mean (range) RUBACE score of 13.8 (8-25) at a mean age of 7.3 years. Segregated by phenotypic categories, there were 39 isolated PBS (76%), six syndromic PBS (12%) and five PBS-plus (10%) cases. The mean RUBACE scores for Woodard categories 1, 2, and 3 were 20.5 (eight patients), 13.8 (25), and 10.6 (17), respectively (P < 0.001). CONCLUSIONS: RUBACE is a practical, organ/system level, phenotyping tool designed to grade PBS severity and categorise patients into isolated PBS, syndromic PBS, and PBS-plus groups. This standardised system will facilitate genotype-phenotype correlations and future prospective multicentre studies assessing medical and surgical treatment outcomes.
Assuntos
Fenótipo , Síndrome do Abdome em Ameixa Seca/classificação , Índice de Gravidade de Doença , Parede Abdominal/patologia , Criança , Pré-Escolar , Criptorquidismo/classificação , Feminino , Humanos , Masculino , Projetos Piloto , Estudos Prospectivos , Ureter/diagnóstico por imagem , Obstrução do Colo da Bexiga Urinária/terapia , Refluxo Vesicoureteral/diagnóstico por imagemRESUMO
Prune Belly Syndrome (PBS) is a congenital multisystem myopathy with mild to lethal severity. While of uncertain etiology, 95% male predominance and familial occurrence suggest a genetic basis. As copy number variations (CNVs) can cause unexplained genetic disorders, we tested for novel CNVs in a large PBS population. We genotyped 21 unrelated PBS patients by high-resolution array comparative genomic hybridization (aCGH) and phenotyped using a novel PBS severity scoring system. Available parents were screened for detected CNV via quantitative PCR (qPCR). We additionally screened for recurrence of identified novel candidate CNVs on 106 PBS probands by qPCR. We identified 10 CNVs in 8 of 21 PBS patients tested (38%). Testing confirmed inheritance from an unaffected biological parent in six patients; parental samples were unavailable in two probands. One candidate CNV includes duplication of the X-chromosome AGTR2 gene, known to function in urinary tract development. Subsequent screening of the larger PBS cohort did not identify any recurrent CNVs. Presence of CNV did not correlate with PBS severity scoring. CNVs were uncommon in this large PBS population, but analysis of identified variants may inform disease pathogenesis and reveal targets for therapeutic intervention for this rare, severe disorder.
Assuntos
Variações do Número de Cópias de DNA/genética , Genética Populacional , Síndrome do Abdome em Ameixa Seca/genética , Adolescente , Feminino , Duplicação Gênica , Testes Genéticos , Humanos , Masculino , Linhagem , Fenótipo , Deleção de SequênciaRESUMO
OBJECTIVE: To prospectively correlate pain duration, red scrotal skin, ultrasound appearance of testis, and intraoperative testis color to future testis atrophy after acute testicular torsion. METHODS: Patients 2 months-18 years old with unilateral acute scrotum were consecutively enrolled in a National Institutes of Health transcutaneous near-infrared spectroscopy study, with a subgroup analysis of the true torsion group. Presence or absence of red scrotal skin, pain duration, testicular heterogeneity on preoperative ultrasound, and intraoperative testis color based on a novel visual chart 5 minutes after detorsion were recorded. All testes underwent orchiopexy regardless of appearance. Percent volume difference between normal and torsed testicles on follow-up ultrasound was compared between patients with and without risk factors. RESULTS: Thirty of 56 patients who had surgical detorsion underwent scrotal ultrasound at a mean of 117 days after surgery. A color of black or hemorrhagic 5 minutes after detorsion, pain duration >12 hours, and heterogeneous parenchyma on preoperative ultrasound were associated with significant testis volume loss in follow-up compared with normal testis. All patients with a black or hemorrhagic testis had >80% volume loss. Erythematous scrotal skin was not significantly associated with smaller affected testis volume in follow-up. CONCLUSION: Based on the high atrophy rate, orchiectomy can be considered for testes that are black or hemorrhagic 5 minutes after detorsion. Pain duration >12 hours and parenchymal heterogeneity on preoperative ultrasound were also associated with testis atrophy. Red scrotal skin was not a reliable predictor of atrophy and should not delay exploration.
Assuntos
Orquiectomia , Orquidopexia/efeitos adversos , Dor/etiologia , Torção do Cordão Espermático/cirurgia , Testículo/patologia , Adolescente , Atrofia/epidemiologia , Atrofia/etiologia , Criança , Pré-Escolar , Cor , Seguimentos , Humanos , Lactente , Masculino , Período Pós-Operatório , Estudos Prospectivos , Medição de Risco , Fatores de Risco , Escroto/diagnóstico por imagem , Espectroscopia de Luz Próxima ao Infravermelho , Torção do Cordão Espermático/complicações , Testículo/diagnóstico por imagem , Fatores de Tempo , UltrassonografiaRESUMO
In this study, we estimate life history parameters and abundance for a protected jaguar population using camera-trap data from a 14-year monitoring program (2002-2015) in Belize, Central America. We investigated the dynamics of this jaguar population using 3,075 detection events of 105 individual adult jaguars. Using robust design open population models, we estimated apparent survival and temporary emigration and investigated individual heterogeneity in detection rates across years. Survival probability was high and constant among the years for both sexes (φ = 0.78), and the maximum (conservative) age recorded was 14 years. Temporary emigration rate for the population was random, but constant through time at 0.20 per year. Detection probability varied between sexes, and among years and individuals. Heterogeneity in detection took the form of a dichotomy for males: those with consistently high detection rates, and those with low, sporadic detection rates, suggesting a relatively stable population of 'residents' consistently present and a fluctuating layer of 'transients'. Female detection was always low and sporadic. On average, twice as many males than females were detected per survey, and individual detection rates were significantly higher for males. We attribute sex-based differences in detection to biases resulting from social variation in trail-walking behaviour. The number of individual females detected increased when the survey period was extended from 3 months to a full year. Due to the low detection rates of females and the variable 'transient' male subpopulation, annual abundance estimates based on 3-month surveys had low precision. To estimate survival and monitor population changes in elusive, wide-ranging, low-density species, we recommend repeated surveys over multiple years; and suggest that continuous monitoring over multiple years yields even further insight into population dynamics of elusive predator populations.
Assuntos
Distribuição Animal , Animais Selvagens , Panthera , Animais , Comportamento Animal , Belize , Feminino , Masculino , Dinâmica PopulacionalRESUMO
PURPOSE: A rapid test for testicular torsion in children may obviate the delay for testicular ultrasound. In this study we assessed testicular tissue percent oxygen saturation (%StO2) measured by transscrotal near infrared spectroscopy as a diagnostic test for pediatric testicular torsion. MATERIALS AND METHODS: This was a prospective comparison to a gold standard diagnostic test study that evaluated near infrared spectroscopy %StO2 readings to diagnose testicular torsion. The gold standard for torsion diagnosis was standard clinical care. From 2013 to 2015 males with acute scrotum for more than 1 month and who were less than 18 years old were recruited. Near infrared spectroscopy %StO2 readings were obtained for affected and unaffected testes. Near infrared spectroscopy Δ%StO2 was calculated as unaffected minus affected reading. The utility of near infrared spectroscopy Δ%StO2 to diagnose testis torsion was described with ROC curves. RESULTS: Of 154 eligible patients 121 had near infrared spectroscopy readings. Median near infrared spectroscopy Δ%StO2 in the 36 patients with torsion was 2.0 (IQR -4.2 to 9.8) vs -1.7 (IQR -8.7 to 2.0) in the 85 without torsion (p=0.004). AUC for near infrared spectroscopy as a diagnostic test was 0.66 (95% CI 0.55-0.78). Near infrared spectroscopy Δ%StO2 of 20 or greater had a positive predictive value of 100% and a sensitivity of 22.2%. Tanner stage 3-5 cases without scrotal edema or with pain for 12 hours or less had an AUC of 0.91 (95% CI 0.86-1.0) and 0.80 (95% CI 0.62-0.99), respectively. CONCLUSIONS: In all children near infrared spectroscopy readings had limited utility in diagnosing torsion. However, in Tanner 3-5 cases without scrotal edema or with pain 12 hours or less, near infrared spectroscopy discriminated well between torsion and nontorsion.
Assuntos
Espectroscopia de Luz Próxima ao Infravermelho , Torção do Cordão Espermático/diagnóstico por imagem , Adolescente , Criança , Pré-Escolar , Edema/complicações , Serviço Hospitalar de Emergência , Humanos , Lactente , Masculino , Valor Preditivo dos Testes , Estudos Prospectivos , Curva ROC , Escroto , Sensibilidade e EspecificidadeRESUMO
PURPOSE: The TWIST (Testicular Workup for Ischemia and Suspected Torsion) score uses urological history and physical examination to assess risk of testis torsion. Parameters include testis swelling (2 points), hard testis (2), absent cremasteric reflex (1), nausea/vomiting (1) and high riding testis (1). While TWIST has been validated when scored by urologists, its diagnostic accuracy among nonurological providers is unknown. We assessed the usefulness of the TWIST score when determined by nonurological nonphysician providers, mirroring emergency room evaluation of acute scrotal pain. MATERIALS AND METHODS: Children with unilateral acute scrotum were prospectively enrolled in a National Institutes of Health clinical trial. After undergoing basic history and physical examination training, emergency medical technicians calculated TWIST score and determined Tanner stage per pictorial diagram. Clinical torsion was confirmed by surgical exploration. All data were captured into REDCap™ and ROC curves were used to evaluate the diagnostic usefulness of TWIST. RESULTS: Of 128 patients (mean age 11.3 years) 44 (13.0 years) had torsion. TWIST score cutoff values of 0 and 6 derived from ROC analysis identified 31 high, 57 intermediate and 40 low risk cases (positive predictive value 93.5%, negative predictive value 100%). CONCLUSIONS: TWIST score assessed by nonurologists, such as emergency medical technicians, is accurate. Low risk patients do not require ultrasound to rule out torsion. High risk patients can proceed directly to surgery, with more than 50% avoiding ultrasound. In the future emergency medical technicians and/or emergency room triage personnel may be able to calculate TWIST score to guide radiological evaluation and immediate surgical intervention at initial assessment long before urological consultation.
Assuntos
Escroto/patologia , Torção do Cordão Espermático/diagnóstico , Testículo/patologia , Adolescente , Criança , Pré-Escolar , Humanos , Masculino , Exame Físico/métodos , Valor Preditivo dos Testes , Estudos Prospectivos , Curva ROC , Encaminhamento e Consulta , Medição de Risco/métodos , Escroto/cirurgia , Torção do Cordão Espermático/cirurgia , Testículo/cirurgia , Ultrassonografia/métodosRESUMO
BACKGROUND: The agents of spotted fevers in Latin America are Rickettsia rickettsii, R. parkeri, Rickettsia sp. strain Atlantic rainforest, and R. massiliae. In Continental Central America, R. rickettsii remains the only known pathogenic tick-borne rickettsia. In the present study, ticks were collected from wild mammals in natural areas of Belize. Besides providing new data of ticks from Belize, we investigated rickettsial infection in some of these ticks. Our results provide ticks harboring rickettsial agents for the first time in Central America. METHODS: Between 2010 and 2015, wild mammals were lived-trapped in the tropical broadleaf moist forests of central and southern Belize. Ticks were collected from the animals and identified to species by morphological and molecular analysis (DNA sequence of the tick mitochondrial 16S RNA gene). Some of the ticks were tested for rickettsial infection by molecular methods (DNA sequences of the rickettsial gltA and ompA genes). RESULTS: A total of 84 ticks were collected from 8 individual hosts, as follows: Amblyomma pacae from 3 Cuniculus paca; Amblyomma ovale and Amblyomma coelebs from a Nasua narica; A. ovale from an Eira Barbara; A. ovale, Amblyomma cf. oblongoguttatum, and Ixodes affinis from a Puma concolor; and A. ovale, A. coelebs, A. cf. oblongoguttatum, and I. affinis from two Panthera onca. Three rickettsial agents were detected: Rickettsia amblyommii in A. pacae, Rickettsia sp. strain Atlantic rainforest in A. ovale, and Rickettsia sp. endosymbiont in Ixodes affinis. CONCLUSIONS: The present study provides unprecedented records of ticks harboring rickettsial agents in the New World. An emerging rickettsial pathogen of South America, Rickettsia sp. strain Atlantic rainforest, is reported for the first time in Central America. Besides expanding the distribution of 3 rickettsial agents in Central America, our results highlight the possible occurrence of Rickettsia sp. strain Atlantic rainforest-caused spotted fever human cases in Belize, since its possible vector, A. ovale, is recognized as one of the most important human-biting ticks in the Neotropical region.
Assuntos
Rickettsia/classificação , Rickettsia/isolamento & purificação , Infestações por Carrapato/epidemiologia , Infestações por Carrapato/parasitologia , Carrapatos/classificação , Carrapatos/microbiologia , Animais , Animais Selvagens , Proteínas de Bactérias/genética , Belize/epidemiologia , Análise por Conglomerados , DNA Mitocondrial/química , DNA Mitocondrial/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Dados de Sequência Molecular , Filogenia , RNA Ribossômico 16S/genética , Rickettsia/genética , Rickettsia rickettsii , Análise de Sequência de DNA , Carrapatos/anatomia & histologia , Carrapatos/genéticaRESUMO
Animal models are key in biomedical research as a proof of concept to study complex processes in a physiological context. Despite the small yet crucial role animals play in fundamental and applied research, the value of animal research is recurrently undermined. Lack of openness and transparency encourages misconceptions, which can have a dramatic negative impact on science and medicine. Research centres should use all available resources to ensure that relevant details about their use of animals in research are readily accessible. More concerted efforts by professional advocacy groups devoted to informing about the benefits of biomedical animal research are also crucial. The European Animal Research Association acts as an umbrella organisation providing support to national advocacy groups and coordinating actions in countries in which no advocacy group exists.
Assuntos
Experimentação Animal , Europa (Continente) , OrganizaçõesRESUMO
The RUN and FYVE domain proteins rabip4 and rabip4' are encoded by RUFY1 and differ in a 108 amino acid N-terminal extension in rabip4'. Their identical C terminus binds rab5 and rab4, but the function of rabip4s is incompletely understood. We here found that silencing RUFY1 gene products promoted outgrowth of plasma membrane protrusions, and polarized distribution and clustering of lysosomes at their tips. An interactor screen for proteins that function together with rabip4' yielded the adaptor protein complex AP-3, of which the hinge region in the ß3 subunit bound directly to the FYVE domain of rabip4'. Rabip4' colocalized with AP-3 on a tubular subdomain of early endosomes and the extent of colocalization was increased by a dominant negative rab4 mutant. Knock-down of AP-3 had an ever more dramatic effect and caused accumulation of lysosomes in protrusions at the plasma membrane. The most peripheral lysosomes were localized beyond microtubules, within the cortical actin network. Our results uncover a novel function for AP-3 and rabip4' in regulating lysosome positioning through an interorganellar pathway.
Assuntos
Complexo 3 de Proteínas Adaptadoras/metabolismo , Endossomos/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Lisossomos/metabolismo , Complexo 3 de Proteínas Adaptadoras/genética , Proteínas Adaptadoras de Transdução de Sinal , Animais , Western Blotting , Linhagem Celular , Membrana Celular/metabolismo , Fibroblastos/citologia , Fibroblastos/metabolismo , Citometria de Fluxo , Células HEK293 , Células HeLa , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Proteína 1 de Membrana Associada ao Lisossomo/metabolismo , Camundongos Mutantes , Microscopia de Fluorescência , Microtúbulos/metabolismo , Ligação Proteica , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , Interferência de RNA , Tetraspanina 30/metabolismo , Proteínas de Transporte Vesicular/metabolismo , Proteínas rab5 de Ligação ao GTP/metabolismoRESUMO
Rab6 is a conserved small GTPase that localizes to the Golgi apparatus and cytoplasmic vesicles and controls transport and fusion of secretory carriers [1]. Another Rab implicated in trafficking from the trans-Golgi to the plasma membrane is Rab8 [2-5]. Here we show that Rab8A stably associates with exocytotic vesicles in a Rab6-dependent manner. Rab8A function is not needed for budding or motility of exocytotic carriers but is required for their docking and fusion. These processes also depend on the Rab6-interacting cortical factor ELKS [1], suggesting that Rab8A and ELKS act in the same pathway. We show that Rab8A and ELKS can be linked by MICAL3, a member of the MICAL family of flavoprotein monooxygenases [6]. Expression of a MICAL3 mutant with an inactive monooxygenase domain resulted in a strong accumulation of secretory vesicles that were docked at the cell cortex but failed to fuse with the plasma membrane, an effect that correlated with the strongly reduced mobility of MICAL3. We propose that the monooxygenase activity of MICAL3 is required to regulate its own turnover and the concomitant remodeling of vesicle-docking protein complexes in which it is engaged. Taken together, the results of our study illustrate cooperation of two Rab proteins in constitutive exocytosis and implicates a redox enzyme in this process.
Assuntos
Exocitose/fisiologia , Oxigenases de Função Mista/fisiologia , Proteínas rab de Ligação ao GTP/fisiologia , Transporte Biológico , Membrana Celular/metabolismo , Vesículas Citoplasmáticas/metabolismo , Células HeLa , Humanos , Fusão de Membrana , Oxigenases de Função Mista/análise , Oxigenases de Função Mista/metabolismo , Oxirredução , Proteínas rab de Ligação ao GTP/análise , Proteínas rab de Ligação ao GTP/metabolismoRESUMO
The endosomal pathway in neuronal dendrites is essential for membrane receptor trafficking and proper synaptic function and plasticity. However, the molecular mechanisms that organize specific endocytic trafficking routes are poorly understood. Here, we identify GRIP-associated protein-1 (GRASP-1) as a neuron-specific effector of Rab4 and key component of the molecular machinery that coordinates recycling endosome maturation in dendrites. We show that GRASP-1 is necessary for AMPA receptor recycling, maintenance of spine morphology, and synaptic plasticity. At the molecular level, GRASP-1 segregates Rab4 from EEA1/Neep21/Rab5-positive early endosomal membranes and coordinates the coupling to Rab11-labelled recycling endosomes by interacting with the endosomal SNARE syntaxin 13. We propose that GRASP-1 connects early and late recycling endosomal compartments by forming a molecular bridge between Rab-specific membrane domains and the endosomal SNARE machinery. The data uncover a new mechanism to achieve specificity and directionality in neuronal membrane receptor trafficking.
Assuntos
Dendritos/metabolismo , Endossomos/metabolismo , Proteínas rab4 de Ligação ao GTP/metabolismo , Animais , Transporte Biológico , Células COS , Proteínas de Transporte/análise , Proteínas de Transporte/metabolismo , Proteínas de Transporte/fisiologia , Chlorocebus aethiops , Dendritos/ultraestrutura , Escherichia coli/genética , Células HeLa , Humanos , Membranas Intracelulares/metabolismo , Membranas Intracelulares/fisiologia , Camundongos , Plasticidade Neuronal , Proteínas Qa-SNARE/metabolismo , Ratos , Receptores de Glutamato/metabolismo , Suínos , Proteínas rab4 de Ligação ao GTP/análise , Proteínas rab4 de Ligação ao GTP/fisiologiaRESUMO
BACKGROUND AND PURPOSE: The prevalence of refractive errors in the general population is between 12 to 18%. The objective of the present study was to determine the refractive errors in healthy children and compare these results in patients with Down syndrome and with cerebral palsy. MATERIAL AND METHOD: The refractive errors in patients between 6 to 12 years without ophthalmologic conditions were studied. The patients were divided into three groups: normal children, children with Down syndrome, and children with cerebral palsy. The results were compared. RESULTS: 258 patient were included: 200 normal children, 29 with Down syndrome, and 29 with cerebral palsy. Ametropia was found in 79.5% in the group of normal children, 93.2% in children with Down syndrome, and in 100% in children with cerebral palsy. Patients requiring optical correction included 30% in normal children, 48.2% in patients with Down syndrome, and 41.1% in the cerebral palsy group. Hyperopia was the most frequent refractive error in all three groups. Most of the refractive errors were mild to moderate. CONCLUSIONS: Refractive errors are more frequent in patients with Down syndrome and cerebral palsy compared with normal children.
RESUMO
The public health workforce has been studied, but not health educators specifically. The development of a Master of Public Health (MPH) in community health education inspired research to illuminate the San Francisco MPH health educator labor market. A time-series survey of an employer panel was conducted between 1995 and 1999 regarding the number of MPH health educators on staff, hiring projections, and importance of selected competencies. In the San Francisco Bay Area, there were 4 MPH health educators per 100,000 persons in 1999. The majority worked in local health departments and community-based organizations. Although hiring was largely replacement in the late 1990s, employers anticipated an increase in hiring from 2000 to 2004. Employers reported that educational preparation was adequate, although preparation in specific competencies, such as bilingual competence, was lacking. These results suggest a favorable labor market for MPH health educators in the San Francisco Bay Area.