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PURPOSE: The gold standard for identification of post-zygotic variants (PZVs) is droplet digital PCR (ddPCR) or high-depth sequencing across multiple tissues types. These approaches are yet to be systematically implemented for monogenic disorders. We developed PZV detection pipelines for correct classification of de novo variants. METHOD: Our pipelines detect PZV in parents (gonosomal mosaicism "pGoM") and children (somatic mosaicism, "M3"). We applied them to research exome sequencing (ES) data from The Australian Cerebral Palsy Biobank (ACPB, n=145 trios) and Simons Simplex Collection (SSC, n=405 families). Candidate mosaic variants were validated using deep amplicon sequencing or ddPCR. RESULTS: 69.2% (M3trio), 63.9% (M3single) and 92.7% (pGoM) of detected variants were validated, with 48.6%, 56.7% and 26.2% of variants respectively meeting strict criteria for mosaicism. In the ACPB, 16.6% of probands and 20.7% of parents had at least one true positive somatic or pGoM variant respectively. A large proportion of PZVs detected in SSC parents (79.8%) and child (94.5%) were not previously reported. We reclassified 3.7-8.0% of germline de novo variants as mosaic. CONCLUSION: Many PZVs were incorrectly classified as germline variants or missed by previous approaches. Systematic application of our pipelines could increase genetic diagnostic rate, improve estimates of recurrence risk in families, and benefit novel disease gene identification.
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OBJECTIVE: To compare strength of left paramedian colopexies using various techniques in equine ex vivo models. STUDY DESIGN: Experimental study. SAMPLE POPULATION: Equine cadavers euthanized for nongastrointestinal pathology (36 specimens derived from 9 horses). METHODS: Colopexies were performed after euthanasia. Suture pattern (horizontal mattress vs. cruciate) and incorporation of dorsal sheath of the rectus abdominis (partial-thickness) versus incorporation of dorsal and ventral sheath of the rectus abdominis (full-thickness) were evaluated. Single cycle load to failure, work to peak load, stiffness, and mode of failure of colopexy tissue constructs were assessed. RESULTS: Mean load to failure of all constructs ranged from 102.26 to 166.38 N. Partial-thickness bites demonstrated a mean load to failure and standard deviation (SD) of 111.91 (35.88) N and 102.26 (30.06) N (p < .05) which was significantly lower than the mean and SD of full-thickness bites (166.3 [72.42] N and 163.21 [51.40 N]), respectively. All full-thickness bites regardless of suture pattern and over half of partial-thickness bites failed at the colonic wall. There was no significant difference in load to failure compared to mode of failure. CONCLUSION: A stronger colopexy was achieved with a full-thickness bite regardless of the suture pattern. The most common mode of failure was the colon wall. CLINICAL SIGNIFICANCE: Incorporating ventral and dorsal fascia of the rectus abdominus provided a stronger colopexy structure, which may necessitate a second incision or subcutaneous palpation of the needle when performing a colopexy. The lateral band of the colon failed in most constructs (77%) regardless of technique, which could weaken the colonic wall and risk colonic rupture.
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Understanding the impact of splicing and nonsense variants on RNA is crucial for the resolution of variant classification as well as their suitability for precision medicine interventions. This is primarily enabled through RNA studies involving transcriptomics followed by targeted assays using RNA isolated from clinically accessible tissues (CATs) such as blood or skin of affected individuals. Insufficient disease gene expression in CATs does however pose a major barrier to RNA based investigations, which we show is relevant to 1,436 Mendelian disease genes. We term these "silent" Mendelian genes (SMGs), the largest portion (36%) of which are associated with neurological disorders. We developed two approaches to induce SMG expression in human dermal fibroblasts (HDFs) to overcome this limitation, including CRISPR-activation-based gene transactivation and fibroblast-to-neuron transdifferentiation. Initial transactivation screens involving 40 SMGs stimulated our development of a highly multiplexed transactivation system culminating in the 6- to 90,000-fold induction of expression of 20/20 (100%) SMGs tested in HDFs. Transdifferentiation of HDFs directly to neurons led to expression of 193/516 (37.4%) of SMGs implicated in neurological disease. The magnitude and isoform diversity of SMG expression following either transactivation or transdifferentiation was comparable to clinically relevant tissues. We apply transdifferentiation and/or gene transactivation combined with short- and long-read RNA sequencing to investigate the impact that variants in USH2A, SCN1A, DMD, and PAK3 have on RNA using HDFs derived from affected individuals. Transactivation and transdifferentiation represent rapid, scalable functional genomic solutions to investigate variants impacting SMGs in the patient cell and genomic context.
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Transdiferenciación Celular , Fibroblastos , Neuronas , Activación Transcripcional , Humanos , Transdiferenciación Celular/genética , Fibroblastos/metabolismo , Fibroblastos/citología , Neuronas/metabolismo , Neuronas/citología , ARN/genética , ARN/metabolismo , Sistemas CRISPR-CasRESUMEN
Clustering Epilepsy (CE) is a neurological disorder caused by pathogenic variants of the Protocadherin 19 (PCDH19) gene. PCDH19 encodes a protein involved in cell adhesion and Estrogen Receptor α mediated-gene regulation. To gain further insights into the molecular role of PCDH19 in the brain, we investigated the PCDH19 interactome in the developing mouse hippocampus and cortex. Combined with a meta-analysis of all reported PCDH19 interacting proteins, our results show that PCDH19 interacts with proteins involved in actin, microtubule, and gene regulation. We report CAPZA1, αN-catenin and, importantly, ß-catenin as novel PCDH19 interacting proteins. Furthermore, we show that PCDH19 is a regulator of ß-catenin transcriptional activity, and that this pathway is disrupted in CE individuals. Overall, our results support the involvement of PCDH19 in the cytoskeletal network and point to signalling pathways where PCDH19 plays critical roles.
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We implicated the X-chromosome THOC2 gene, which encodes the largest subunit of the highly-conserved TREX (Transcription-Export) complex, in a clinically complex neurodevelopmental disorder with intellectual disability as the core phenotype. To study the molecular pathology of this essential eukaryotic gene, we generated a mouse model based on a hypomorphic Thoc2 exon 37-38 deletion variant of a patient with ID, speech delay, hypotonia, and microcephaly. The Thoc2 exon 37-38 deletion male (Thoc2Δ/Y) mice recapitulate the core phenotypes of THOC2 syndrome including smaller size and weight, and significant deficits in spatial learning, working memory and sensorimotor functions. The Thoc2Δ/Y mouse brain development is significantly impacted by compromised THOC2/TREX function resulting in R-loop accumulation, DNA damage and consequent cell death. Overall, we suggest that perturbed R-loop homeostasis, in stem cells and/or differentiated cells in mice and the patient, and DNA damage-associated functional alterations are at the root of THOC2 syndrome.
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Discapacidad Intelectual , Factores de Transcripción , Humanos , Masculino , Ratones , Animales , Factores de Transcripción/metabolismo , Estructuras R-Loop , Transporte Activo de Núcleo Celular , Discapacidad Intelectual/genética , Daño del ADN , Fenotipo , ARN Mensajero/metabolismoAsunto(s)
Rotura de la Aorta , Enfermedades de los Caballos , Derrame Pericárdico , Caballos , Animales , Femenino , Derrame Pericárdico/diagnóstico , Derrame Pericárdico/veterinaria , Rotura de la Aorta/veterinaria , Enfermedades de los Caballos/diagnóstico , Dolor Abdominal/diagnóstico , Dolor Abdominal/etiología , Dolor Abdominal/veterinaria , Hemotórax/veterinariaRESUMEN
Aicardi Syndrome (AIC) is a rare neurodevelopmental disorder recognized by the classical triad of agenesis of the corpus callosum, chorioretinal lacunae and infantile epileptic spasms syndrome. The diagnostic criteria of AIC were revised in 2005 to include additional phenotypes that are frequently observed in this patient group. AIC has been traditionally considered as X-linked and male lethal because it almost exclusively affects females. Despite numerous genetic and genomic investigations on AIC, a unifying X-linked cause has not been identified. Here, we performed exome and genome sequencing of 10 females with AIC or suspected AIC based on current criteria. We identified a unique de novo variant, each in different genes: KMT2B, SLF1, SMARCB1, SZT2 and WNT8B, in five of these females. Notably, genomic analyses of coding and non-coding single nucleotide variants, short tandem repeats and structural variation highlighted a distinct lack of X-linked candidate genes. We assessed the likely pathogenicity of our candidate autosomal variants using the TOPflash assay for WNT8B and morpholino knockdown in zebrafish (Danio rerio) embryos for other candidates. We show expression of Wnt8b and Slf1 are restricted to clinically relevant cortical tissues during mouse development. Our findings suggest that AIC is genetically heterogeneous with implicated genes converging on molecular pathways central to cortical development.
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Síndrome de Aicardi , Masculino , Femenino , Animales , Ratones , Síndrome de Aicardi/genética , Pez Cebra/genética , Mapeo Cromosómico , Genes Ligados a X/genética , BioensayoRESUMEN
OBJECTIVE: To assess oral buccal microcirculation by hand-held videomicroscopy in horses during colic surgery, comparing microcirculation values with macrocirculatory parameters and with those of healthy elective surgical horses. STUDY DESIGN: Clinical prospective study. ANIMALS: Client-owned horses (nine in the colic group; 11 in the elective group). METHODS: In the colic group, buccal mucosal side stream dark-field microscopy (DFM) videos, cardiac output (CO), mean arterial pressure (MAP), and lactate were obtained at three timepoints under general anesthesia (30, 90, and 150 min after induction). Video analysis was used to determine total vessel density, proportion of perfused vessels, perfused vessel density, and heterogeneity index. Dark-field microscopy videos, MAP, and lactate were obtained at a single timepoint under general anesthesia (45 min after induction) in the elective group. RESULTS: There were no differences in microcirculatory parameters between colic and elective horses, nor was there a difference across timepoints in the colic group. There was a weak negative correlation between microvascular parameters and CO (rho = -0.23). CONCLUSION: The colic group did not have decreased microcirculation in comparison with the healthy elective group. Dark-field microscopy did not correlate well with macrocirculatory parameters in the colic group. IMPACT: Dark-field microscopy may not be a sensitive enough indicator to detect differences in microcirculation between colic and elective groups. The lack of difference in microcirculation may be due to sample size, probe location, or variation in disease severity.
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Cólico , Enfermedades de los Caballos , Animales , Cólico/cirugía , Cólico/veterinaria , Procedimientos Quirúrgicos Electivos/veterinaria , Enfermedades de los Caballos/cirugía , Caballos/cirugía , Ácido Láctico , Microcirculación , Estudios ProspectivosRESUMEN
BACKGROUND: The COVID-19 pandemic accelerated the use and acceptance of telemedicine. Simultaneously, emergency departments (EDs) have experienced increased ED boarding. With this acceptance of telemedicine and the weighty increase in patient boarding, we proposed the innovative Virtual First (VF) program to leverage emergency medicine clinicians' (EMCs) ability to triage patients. VF seeks to reduce unnecessary ED visits by connecting patients with EMCs prior to seeking in-person care rather than using traditional ED referral systems. OBJECTIVE: The goal of this study is to investigate how patients' access to EMCs from home via the establishment of VF changed how patients sought care for acute care needs. METHODS: VF is a synchronous virtual video visit at a tertiary care academic hospital. VF was staffed by EMCs and enabled full management of patient complaints or, if necessary, referral to the appropriate level of care. Patients self-selected this service as an alternative to seeking in-person care at a primary care provider, urgent care center, or ED. A postvisit convenience sample survey was collected through a phone SMS text message or email to VF users. This is a cross-sectional survey study. The primary outcome measure is based on responses to the question "How would you have sought care if a VF visit was not available to you?" Secondary outcome measures describe valued aspects and criticisms. Results were analyzed using descriptive statistics. RESULTS: There were 3097 patients seen via VF from July 2021 through May 2022. A total of 176 (5.7%) patients completed the survey. Of these, 87 (49.4%) would have sought care at urgent care centers if VF had not been available. There were 28 (15.9%) patients, 26 (14.8%) patients, and 1 (0.6%) patient that would have sought care at primary care providers, EDs, or other locations, respectively. Interestingly, 34 (19.3%) patients would not have sought care. The most valued aspect of VF was receiving care in the comfort of the home (n=137, 77.8%). For suggested improvements, 58 (33%) patients most commonly included "Nothing" as free text. CONCLUSIONS: VF has the potential to restructure how patients seek medical care by connecting EMCs with patients prior to ED arrival. Without the option of VF, 64.2% (113/177) of patients would have sought care at an acute care facility. VF's innovative employment of EMCs allows for acute care needs to be treated virtually if feasible. If not, EMCs understand the local resources to better direct patients to the appropriate site. This has the potential to substantially decrease patient costs because patients are given the appropriate destination for in-person care, reducing the likelihood of the need for transfer and multiple ED visits.
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COVID-19 , Medicina de Emergencia , Telemedicina , Humanos , Estudios Transversales , Pandemias , Servicio de Urgencia en HospitalRESUMEN
BACKGROUND: Firearm injury is the leading cause of death in children and prevention is the most effective method to reduce severe morbidity and mortality. Injury prevention programs have used community firearm lock giveaway events to promote safe firearm storage practices. The locks are generally simple and inexpensive devices suitable for mass distribution but may not possess the owner-desired attributes for use in the home. Because data on owner preferences for firearm lock type is lacking, we conducted a community survey to inform firearm safety outreach efforts. METHODS: We performed an anonymous cross-sectional survey at a large community fair. We elicited responses regarding reasons for firearm ownership, current storage practices, and preferences for firearm storage devices. Participants were offered a choice of a free trigger lock or cable lock and education on its use. RESULTS: Two-hundred and sixty-seven of 394 (67.7%) respondents reported firearm ownership, with 64.8% reporting children in the home regularly. Most (60.7%) owned handguns and cited personal protection as the main reason for ownership (88.4%). The ability to store the firearm loaded and the need for rapid access were identified as the main storage considerations. Respondents preferred trigger locks over cable locks at a rate of almost 2:1. CONCLUSIONS: The majority of firearm owners had handguns for self-defense. Owners preferred simple locking mechanisms that allowed the firearm to remain loaded. The pragmatic pediatric injury prevention program will include firearm owners' preferences when considering which lock to purchase and distribute during firearm injury prevention programs. LEVEL OF EVIDENCE: IV, Epidemiological.
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Armas de Fuego , Heridas por Arma de Fuego , Humanos , Niño , Heridas por Arma de Fuego/prevención & control , Estudios Transversales , Encuestas y Cuestionarios , Propiedad , SeguridadRESUMEN
Chemical genetic approaches have had a transformative impact on discovery of drug targets for malaria but have primarily been used for parasite targets. To identify human pathways required for intrahepatic development of parasite, we implemented multiplex cytological profiling of malaria infected hepatocytes treated with liver stage active compounds. Some compounds, including MMV1088447 and MMV1346624, exhibited profiles similar to cells treated with nuclear hormone receptor (NHR) agonist/antagonists. siRNAs targeting human NHRs, or their signaling partners identified eight genes that were critical for Plasmodium berghei infection. Knockdown of NR1D2, a host NHR, significantly impaired parasite growth by downregulation of host lipid metabolism. Importantly, treatment with MMV1088447 and MMV1346624 but not other antimalarials, phenocopied the lipid metabolism defect of NR1D2 knockdown. Our data underlines the use of high-content imaging for host-cellular pathway deconvolution, highlights host lipid metabolism as a drug-able human pathway and provides new chemical biology tools for studying host-parasite interactions.
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Malaria , Parásitos , Animales , Humanos , Hepatocitos/metabolismo , Hígado/metabolismo , Malaria/tratamiento farmacológico , Malaria/metabolismo , Plasmodium berghei/genéticaRESUMEN
OBJECTIVE: To report the prevalence and risk factors for incisional morbidities in late pregnant and nonpregnant/early pregnant control mares following colic surgery. DESIGN: Multicenter, retrospective, cohort study from January 2014 to December 2019. SETTING: Two university teaching hospitals and 1 private referral center. ANIMALS: Five hundred and seventy-nine fillies and mares ≥2 years old that underwent celiotomy. Pregnant mares (n = 54) were >240 days in gestation from the last known breeding date and were compared to control females (n = 525) undergoing colic surgery. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Morbidity rates were not different between mare groups with 56% of pregnant mares and 51% of control mares reporting at least 1 morbidity. Incisional swelling was the most common reported complication in both groups. Incisional swelling was associated with shorter hospital stays (odds ratio [OR], 0.18; P < 0.01), and drainage was associated with a longer hospital stay (OR, 1.27; P ≤ 0.01) and with use of an abdominal bandage (OR, 4.4; P < 0.01). Herniation was associated with hypercapnia under anesthesia (OR, 1.1; P = 0.048), previous abdominal surgery (OR, 8.3; P = 0.003), and with use of an abdominal bandage (OR, 56; P = 0.006). Body wall dehiscence was associated with longer hospital stay (OR, 1.2; P < 0.01). Nonsurvival was higher in pregnant mares (13%) compared to control mares (5%; P = 0.02). CONCLUSIONS: The prevalence of incisional morbidities did not differ between pregnant and control mares undergoing colic surgery. Several factors were associated with incisional morbidities, including the duration of surgery and anesthesia, anesthetic variables, abdominal bandage use, previous ventral abdominal incision, and longer duration of hospitalization.
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Cólico , Enfermedades de los Caballos , Embarazo , Caballos , Animales , Femenino , Cólico/epidemiología , Cólico/cirugía , Cólico/veterinaria , Prevalencia , Estudios Retrospectivos , Estudios de Cohortes , Enfermedades de los Caballos/epidemiología , Enfermedades de los Caballos/cirugía , Enfermedades de los Caballos/etiología , MorbilidadRESUMEN
This article discusses the unequal impact of Covid-19 on the lives of the children of survivors of modern slavery, child victims of exploitation and children at risk of exploitation in the UK. It draws on research that has analysed the risks and impacts of Covid-19 on victims and survivors of modern slavery. It explores how pandemic responses may have hindered these children's rights to education, food, safety, development and participation and representation in legal processes. It suggests that the pandemic should be used as an impetus to address inequalities that existed pre-Covid-19 and those that have been exacerbated by it.
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BACKGROUND: Increased intraabdominal pressure, termed intraabdominal hypertension (IAH), is reported as an independent cause of morbidity and mortality in the human ICU but, until recently, has been rarely described in veterinary species outside of experimental models. Failure to identify severe IAH leads to organ dysfunction, termed abdominal compartment syndrome, and rapidly becomes fatal without therapeutic intervention. Although the veterinary community has been slow to address the concept of IAH and associated comorbidities, recent companion and large animal case series and experimental studies suggest IAH may also be common in veterinary species and correlates well with risk factors and grading systems already described in the human literature. PATHOPHYSIOLOGY: Increasing abdominal pressures exert deleterious local effects through visceral ischemia and reperfusion injury as well as systemic effects on the cardiovascular, pulmonary, renal, and central nervous systems. Even mild grades of IAH increase systemic vascular resistance, impede venous return, increase pulmonary wedge pressure, and decrease pulmonary function. More severe grades cause azotemia, oliguria, decreased coronary blood flow, hypoxia, increased intracranial pressure, and death. IMPORTANCE: Many of the common diseases in veterinary patients are associated with IAH, including gastric dilatation-volvulus, colon volvulus, closed pyometra, hemoperitoneum, ascites, uroperitoneum, and hydrops. Monitoring of the veterinary patient is difficult, but several experimental studies validate both the presence of IAH and the ability to monitor abdominal pressures in large and small animal species. Moreover, prompt recognition of IAH and subsequent treatment is feasible in the veterinary ICU. KEY POINTS: Increasing abdominal pressures exert deleterious local effects through visceral ischemia and reperfusion injury as well as systemic effects on the cardiovascular, pulmonary, renal, and central nervous systems. Increases in central venous pressure, systemic vascular resistance, pulmonary wedge pressure, and a decreased cardiac output by way of both decreased preload and increased afterload have been documented as a result of intraabdominal hypertension (IAH). Direct diagnosis of IAH is achieved by blind or ultrasound-guided abdominal needle puncture attached to aâ¯waterâ¯manometerâ¯or direct pressure monitoring transducer. Transvesicular measurement of intraabdominal pressure (IAP) is relatively noninvasive, and many patients that would benefit from rapid diagnosis of IAH and abdominal compartment syndrome already have indwelling bladder catheters. Recommendations for interventions are based on the assigned grade of IAH (mild, moderate, severe). KEY POINTS: If IAH is strongly suspected or diagnosed, abdominal wall compliance may be improved through judicious use of neuromuscular blockers and sedation. Decompression, either minimally invasive or surgical, is absolutely recommended for IAPs consistently above 20 mm Hg, especially in the presence of signs attributed to secondary organ dysfunction.
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Traumatismos Abdominales , Anestesia , Hipertensión Intraabdominal , Traumatismos Abdominales/veterinaria , Anestesia/veterinaria , Animales , Cuidados Críticos , Humanos , Hipertensión Intraabdominal/terapia , Hipertensión Intraabdominal/veterinaria , Factores de RiesgoRESUMEN
BACKGROUND: Cecal or colonic gas tympany of any cause may result in increased intraabdominal pressure, causing a significant decrease in venous return and cardiac output. Trocarization of the large colon or cecum in the event of large intestinal tympany may resolve gas distension and accompanying increased intraabdominal pressures sufficiently enough to promote resolution of a displaced large colon. Furthermore, trocarization of the medical colic may decrease morbidity and mortality associated with severe intraabdominal hypertension. DESCRIPTION: This how-to description details the technique of transcutaneous trocarization of the large colon and cecum in the standing horse using a 14-gauge catheter. SUMMARY: Trocarization is not a substitute for surgery in the severely gas-distended painful horse exhibiting signs of colic that require surgery; however, it is a viable medical therapy with minimal risk that alleviates gas distention and may assist with correction of a colonic displacement in the horse. KEY POINTS: The majority of colic episodes involving the large colon can be managed medically without surgical intervention. Auscultation and percussion of the paralumbar fossa area will solicit an audible "ping" (gas/fluid interface), thus targeting the appropriate site for trocarization. Percutaneous trocarization allows for rapid controlled decompression of colonic or cecal tympany, permitting improvement of both ventilation and perfusion. In situations where a displacement or primary colonic or cecal tympany is suspected, trocarisation is deemed successful if the horse has resolution of colic signs and the physical examination parameters return to normal. Trocarization may be clinically advantageous by requiring less pain medication, decreased time in the hospital, decreased costs, and avoidance of surgical intervention; yet, this therapy should not take place of surgical intervention if deemed necessary.
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Enfermedades del Ciego , Cólico , Enfermedades de los Caballos , Animales , Enfermedades del Ciego/veterinaria , Ciego , Cólico/veterinaria , Colon/cirugía , Enfermedades de los Caballos/cirugía , CaballosRESUMEN
OBJECTIVES: Children with hemophilia have the usual childhood risk of falls and head trauma. Head computed tomographies (HCTs) are fast, detailed, and readily available, but increased radiation exposure in the pediatric population is now recognized as causing increased brain malignancy. By examining the incidence of intracranial cerebral hemorrhage in this population, we will be able to weigh risks and benefits of HCT use more accurately. METHODS: Using a retrospective chart review, we examined past medical records of pediatric patients, aged 0 to 15 years, with hemophilia presenting to 1 academic medical center. Primary outcomes included number of head CTs ordered, total and per patient over the years studied, and the incidence of positive findings, as defined by presence of blood products as documented by radiologist final read/interpretation. RESULTS: The mean number of head CTs per child was 2.5 (range, 1-10). None of the HCT scans were read as intracranial cerebral hemorrhage, and none of the patients had findings that lead to neurosurgical intervention. In a sensitivity analysis, applying Pediatric Emergency Care Applied Research Network head injury criteria, 11 HCT scans would be ordered for a reduction of 80 HCTs, or a decrease of 2 HCT scans per child. No incidence of intracranial cerebral hemorrhage would have been missed. CONCLUSIONS: Our findings suggest that in the child with hemophilia and a history of minor head trauma, exposure to the radiation of a HCT based on the diagnosis of hemophilia alone may not be necessary but that imaging decisions need to be made in conjunction with clinical examination findings and neurologic status.
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Traumatismos Craneocerebrales , Hemofilia A , Exposición a la Radiación , Niño , Traumatismos Craneocerebrales/complicaciones , Traumatismos Craneocerebrales/diagnóstico por imagen , Traumatismos Craneocerebrales/epidemiología , Hemofilia A/complicaciones , Hemofilia A/diagnóstico por imagen , Hemofilia A/epidemiología , Humanos , Estudios Retrospectivos , Tomografía Computarizada por Rayos XRESUMEN
BACKGROUND: A pediatric emergency medicine and critical care fellowship was recently developed in Kenya through the University of Nairobi/Kenyatta National Hospital and AIC Kijabe Hospital. As part of this training, a week-long trauma and emergency medicine course was developed with emphasis on trauma and emergency medicine procedures. Given limited resources, we developed a course with simulation of procedures centered around utilization of a goat cadaver. OBJECTIVE: The aim of the study was to describe fellow and faculty experiences and perspectives when using a goat cadaver to teach emergency medicine procedures by simulation in Kijabe, Kenya. METHODS: A 5-day course was given to 2 fellows with a variety of didactics and simulations after which fellows completed a questionnaire to rate their satisfaction with the content and teaching effectiveness. RESULTS: The course was rated very highly, with an average content satisfaction score of 4.5 5 and average teaching effectiveness score of 4.4 of 5. Qualitative faculty feedback was positive, with specific learnings allowing ongoing adaptation of this model. CONCLUSIONS: A goat cadaver is a cost-effective resource not often considered that can be adequately used to teach several emergency medicine skills by simulation.
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Medicina de Emergencia , Cabras , Animales , Cadáver , Análisis Costo-Beneficio , Curriculum , Medicina de Emergencia/educación , Becas , Humanos , KeniaRESUMEN
BACKGROUND: Incisional complications are a common cause of morbidity following laparotomy. Although uncommon, acute abdominal dehiscence (AAD) is a potentially fatal post-operative complication. However, few AAD cases are described in the literature. OBJECTIVES: To describe common features of cases of AAD following ventral midline laparotomy, management and outcomes. STUDY DESIGN: Retrospective case series. METHODS: Hospital records of horses that underwent a ventral midline laparotomy at nine hospitals in the UK, Ireland and USA over a 10-year period (2009-2019) were reviewed. Data were collected for pre-, intra- and post-operative factors that were considered relevant. Descriptive statistical analysis was performed. RESULTS: A total of 63 cases of AAD were identified. AAD occurred due to tearing of sutures through the linea alba or rupture of the body wall adjacent to the suture line in 46 horses (73%). AAD occurred at a median of 5 days (0.5-70 days) post-operatively and broodmares accounted for 25% of the cases (n = 16). Surgical site infection developed prior to AAD in 28 horses (44%); leakage of peritoneal fluid occurred in 5% of horses prior to AAD being identified. Surgical repair was performed in 27 horses (43%), 10 (16%) were treated conservatively and 26 (41%) were euthanised immediately. Repair was most frequently performed using suture (n = 14), wire (n = 5) or a combination (n = 5). Overall survival to hospital discharge was 39% (24/63). Where surgical repair was performed, 15 horses (56%) survived to hospital discharge; 9 horses (90%) managed conservatively survived to hospital discharge. MAIN LIMITATIONS: Follow-up was not performed for all cases following hospital discharge and some data were incompletely recorded in hospital files. CONCLUSIONS: Previously stated causative factors for AAD were not consistent features in the present study. Surgical site infection following laparotomy and pregnant or early post-partum mares may be important risk factors for AAD and warrant further investigation.
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PCDH19 is a nonclustered protocadherin molecule involved in axon bundling, synapse function, and transcriptional coregulation. Pathogenic variants in PCDH19 cause infantile-onset epilepsy known as PCDH19-clustering epilepsy or PCDH19-CE. Recent advances in DNA-sequencing technologies have led to a significant increase in the number of reported PCDH19-CE variants, many of uncertain significance. We aimed to determine the best approaches for assessing the disease relevance of missense variants in PCDH19. The application of the American College of Medical Genetics and Association for Molecular Pathology (ACMG-AMP) guidelines was only 50% accurate. Using a training set of 322 known benign or pathogenic missense variants, we identified MutPred2, MutationAssessor, and GPP as the best performing in silico tools. We generated a protein structural model of the extracellular domain and assessed 24 missense variants. We also assessed 24 variants using an in vitro reporter assay. A combination of these tools was 93% accurate in assessing known pathogenic and benign PCDH19 variants. We increased the accuracy of the ACMG-AMP classification of 45 PCDH19 variants from 50% to 94%, using these tools. In summary, we have developed a robust toolbox for the assessment of PCDH19 variant pathogenicity to improve the accuracy of PCDH19-CE variant classification.
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Cadherinas , Epilepsia , Cadherinas/genética , Humanos , Mutación Missense , Protocadherinas , Análisis de Secuencia de ADNRESUMEN
OBJECTIVE: To identify the causative gene in a large unsolved family with genetic epilepsy with febrile seizures plus (GEFS+), we sequenced the genomes of family members, and then determined the contribution of the identified gene to the pathogenicity of epilepsies by examining sequencing data from 2,772 additional patients. METHODS: We performed whole genome sequencing of 3 members of a GEFS+ family. Subsequently, whole exome sequencing data from 1,165 patients with epilepsy from the Epi4K dataset and 1,329 Australian patients with epilepsy from the Epi25 dataset were interrogated. Targeted resequencing was performed on 278 patients with febrile seizures or GEFS+ phenotypes. Variants were validated and familial segregation examined by Sanger sequencing. RESULTS: Eight previously unreported missense variants were identified in SLC32A1, coding for the vesicular inhibitory amino acid cotransporter VGAT. Two variants cosegregated with the phenotype in 2 large GEFS+ families containing 8 and 10 affected individuals, respectively. Six further variants were identified in smaller families with GEFS+ or idiopathic generalized epilepsy (IGE). CONCLUSION: Missense variants in SLC32A1 cause GEFS+ and IGE. These variants are predicted to alter γ-aminobutyric acid (GABA) transport into synaptic vesicles, leading to altered neuronal inhibition. Examination of further epilepsy cohorts will determine the full genotype-phenotype spectrum associated with SLC32A1 variants.