Heterogeneity of Wnt1-Cre-marked and Pax2-Cre-marked first branchial arch cranial neural crest cells in mice.

OBJECTIVES: This study aimed to explore the heterogeneity and gene ontology of Wnt1-Cre-marked and Pax2-Cre-marked first branchial arch cranial neural crest cells (CNCs) in mice. METHODS: The embryos of Wnt1-Cre;R26RmTmG and Pax2-Cre;R26RmTmG at embryonic day (E)8.0-E9.25 were collected for histological observation. We performed immunostaining to compare green fluorescent protein (GFP)-positive CNCs in Pax2-Cre;R26RAi9 and Wnt1-Cre;R26RAi9 mice at E15.5. Single-cell RNA sequencing (scRNA-seq) was used to analyze the first branchial arch GFP-positive CNCs from Wnt1-Cre;R26RmTmG and Pax2-cre;R26RmTmGmice at E10.5. Real time fluorescence quantitative polymerase chain reaction (q-PCR) was performed to validate the differential genes. RESULTS: Wnt1-Cre-marked and Pax2-Cre-marked CNCs migrated from the neural plateto first and second branchial arches and to the first branchial arch, respectively, at E8.0. Although Wnt1-Cre-marked and Pax2-Cre-marked CNCs were found mostly in cranial-facial tissues, the former had higher expression in palate and tongue. The results of scRNA-seq showed that Pax2-Cre-marked CNCs specifically contributed to osteoblast differentiation and ossification, while Wnt1-Cre-marked CNCs participated in limb development, cell migration, and ossification. The q-PCR data also confirmed the results of gene ontology analysis. CONCLUSIONS: Pax2-Cre mice are perfect experimental animal models for research on first branchial arch CNCs and derivatives in osteoblast differentiation and ossification.

Abnormalities in pharyngeal arch-derived structures in SATB2-associated syndrome.

SATB2-associated syndrome (SAS, glass syndrome, OMIM#612313) is a neurodevelopmental autosomal dominant disorder with frequent craniofacial abnormalities including palatal and dental anomalies. To assess the role of Satb2 in craniofacial development, we analyzed mutant mice at different stages of development. Here, we show that Satb2 is broadly expressed in early embryonic mouse development including the mesenchyme of the second and third arches. Satb2-/- mutant mice exhibit microglossia, a shortened lower jaw, smaller trigeminal ganglia, and larger thyroids. We correlate these findings with the detailed clinical phenotype of four individuals with SAS and remarkable craniofacial phenotypes with one requiring mandibular distraction in childhood. We conclude that the mouse and patient data presented support less well-described phenotypic aspects of SAS including mandibular morphology and thyroid anatomical/functional issues.

Microtia, Branchial Cleft Fistula, and Tetralogy of Fallot: A Possible Association.

When searching over associations between congenital ear abnormalities, especially microtia and affiliated deformities like cleft lip or palate and congenital heart diseases, some clinical analysis and genetic theories are found. A 10-year-old boy sent to the plastic surgery hospital was puzzled by a congenital anterior auricular fistula with fluid trace for more than 9 years. The preoperative diagnoses were branchial cleft fistula and congenital left ear deformity with postoperation of TOF. By browsing over studies on genetic concerns and clinical performance, it may be attributed to a possible association between microtia, branchial cleft fistula, and tetralogy of Fallot, though whose fundamental mechanisms remain concerned.

Dyrk1a is required for craniofacial development in Xenopus laevis.

Loss of function variations in the dual specificity tyrosine-phosphorylation-regulated kinase 1 A (DYRK1A) gene are associated with craniofacial malformations in humans. Here we characterized the effects of deficient DYRK1A in craniofacial development using a developmental model, Xenopus laevis. Dyrk1a mRNA and protein were expressed throughout the developing head and both were enriched in the branchial arches which contribute to the face and jaw. Consistently, reduced Dyrk1a function, using dyrk1a morpholinos and pharmacological inhibitors, resulted in orofacial malformations including hypotelorism, altered mouth shape, slanted eyes, and narrower face accompanied by smaller jaw cartilage and muscle. Inhibition of Dyrk1a function resulted in misexpression of key craniofacial regulators including transcription factors and members of the retinoic acid signaling pathway. Two such regulators, sox9 and pax3 are required for neural crest development and their decreased expression corresponds with smaller neural crest domains within the branchial arches. Finally, we determined that the smaller size of the faces, jaw elements and neural crest domains in embryos deficient in Dyrk1a could be explained by increased cell death and decreased proliferation. This study is the first to provide insight into why craniofacial birth defects might arise in humans with variants of DYRK1A.

Pulsative venous return from the branchial vessels to the heart of the bivalve Mytilus galloprovincialis supports the constant-volume mechanism.

In bivalves and gastropods, ventricle contraction causes a negative pressure in the auricles and increases venous return from the afferent oblique vein (AOV): the constant-volume (CV) mechanism. The flow in the AOV should be a pulsative flow synchronized with the ventricular contraction. The flow in the heart and adjacent vessels of Mytilus galloprovincialis were measured by magnetic resonance imaging to confirm this hypothesis. Under a regular heartbeat, pulsative flows in the AOV and branchial vessels (BVs) were almost completely synchronized with the flow in the aorta, while filling of the ventricle was in the opposite phase. Flows in the BVs were directed to the posterior direction, and a pair of BVs in the gill axes (the efferent BVs) were connected to the AOV. Based on the images of the whole pathway of the AOV in an oblique slice, we confirmed that haemolymph flow was evoked from the efferent BVs and flow into the ventricle via the auricle was completed in a single heartbeat. Therefore, the walls of the AOV and BVs could resist negative transmural pressure caused by the ventricular contraction. In conclusion, the auricle, the AOV and the BVs, including the gill filaments, act as a suction pump. The pulsative venous return is driven by the negative pressure of the AOV as in the CV mechanism, and the negative pressure in the efferent BVs could draw haemolymph from the sinus via the gill and the afferent BVs. Therefore, Mytilus can start and stop its heartbeat as necessary.

Congenital midline cervical cleft: Management of a case series and literature review.

Congenital midline cervical cleft is a rare anomaly classified as a malformation of the branchial arches and represents less than 2% of congenital cervical malformations. Its clinical presentation involves cervical midline deformities: cephalic nodular lesion, linear groove with atrophic surface, and/or caudal sinus. Other midline alterations of variable complexity may also be present. Early treatment allows for avoiding long-term complications. Based on our experience in four clinical cases, a performed literature search on the topic in the last twenty years, and subsequent discussion of the employed surgical approaches, we included 150 reported cases in our review. Correct diagnosis and early treatment with complete removal of the fibrous midline band is paramount to avoid patient complaints until adolescence or adulthood.

The congenital sternoclavicular sinus in children: Our experience of 56 cases.

OBJECTIVES: The objective of this study is to explore the clinical diagnosis and treatment of the congenital sternoclavicular sinus in the region of sternoclavicular joint, providing valuable insights for clinical practice in children. METHODOLOGY: A retrospective review of case notes was conducted for all patients treated for congenital sternoclavicular sinus at a tertiary care pediatric otolaryngology practice from January 2022 to September 2023.This review analyzed the clinical presentations, imaging examination, pathological examination, and treatment. RESULTS: A total of 56 patients with congenital sternoclavicular sinus were included in the study, with a mean age of 4.10 ± 2.57 years old. Among these patients, the sinuses were located on the left side in 43 cases,on the right side in 12 cases,and bilaterally in one case.While two patients never experienced inflammation, 54 patients did, and 45 of them underwent incision and drainage before excision. Imaging examination, including ultrasonography and computerized tomography (CT) scans, revealed characteristics such as thickening of subcutaneous tissue,subcutaneous tubular structures or tubercles in front of the clavicle,and irregularly shaped lesions with clear borders and uneven internal low echo, Pathological examination confirmed that the sinuses were lined by squamous epithelium in all patients. CONCLUSIONS: The congenital sternoclavicular sinus may be a remnant of the fourth branchial cleft on the skin side.Complete resection of the sinus during the quiescent phase of inflammation is the recommended curative treatment to prevent recurrent infection.

In vitro induction of patterned branchial arch-like aggregate from human pluripotent stem cells.

Early patterning of neural crest cells (NCCs) in the craniofacial primordium is important for subsequent development of proper craniofacial structures. However, because of the complexity of the environment of developing tissues, surveying the early specification and patterning of NCCs is difficult. In this study, we develop a simplified in vitro 3D model using human pluripotent stem cells to analyze the early stages of facial development. In this model, cranial NCC-like cells spontaneously differentiate from neural plate border-like cells into maxillary arch-like mesenchyme after a long-term culture. Upon the addition of EDN1 and BMP4, these aggregates are converted into a mandibular arch-like state. Furthermore, temporary treatment with EDN1 and BMP4 induces the formation of spatially separated domains expressing mandibular and maxillary arch markers within a single aggregate. These results suggest that this in vitro model is useful for determining the mechanisms underlying cell fate specification and patterning during early facial development.

Sonographic Findings of Cervical Chondrocutaneous Branchial Remnants-A Comparison With Dermal Lesions/Cysts and a Literature Review: A Pilot Study.

OBJECTIVES: Cervical chondrocutaneous branchial remnants (CCBRs) and dermal lesions, such as epidermoid cysts or brachial anomalies, including lateral cervical cysts/sinuses or dermal sinuses of anterior chest lesions, are usually located at the lower neck at the anterior or posterior border of the sternocleidomastoid muscle (SCM). We aimed to demonstrate the usefulness of ultrasonography in the differential diagnosis and evaluation of CCBRs. METHODS: We evaluated 22 lesions of 20 pediatric patients, classified into CCBR and dermal lesion groups. We used Fisher's exact test to evaluate differences between these groups in terms of lesion shape (low-echoic mass- or tubular-like), whether the lesion was adjacent to/in contact with the SCM or not, and the presence or absence of a concave SCM caused by the lesion. RESULTS: Of the 22 lesions, 8 were CCBRs, and 14 were dermal lesions. We found a significant difference in the presence/absence of adjacency to or contact with the SCM (presence/absence of adjacency to or contact with the SCM in CCBRs vs that in dermal lesions: 6/2 vs 1/13, P = .002) and presence/absence of lesion-induced concavity of the SCM (presence/absence of lesion-induced concavity of the SCM in CCBRs vs that in dermal lesions: 3/5 vs 0/14, P = .036). The lesion shape (low-echoic mass-like/tubular-like lesions) did not significantly differ between the two study groups (low-echoic mass-like/tubular-like lesions in CCBRs vs that in dermal lesions: 5/3 vs 11/6, P = .624). CONCLUSIONS: CCBRs have a strong association with the SCM. These sonographic findings may be useful in the differential diagnosis of dermal cervical lesions.

Phylogenetic relationships and description of two new species of Diapoma (Characidae: Stevardiinae) from the La Plata River basin

Herein we describe two new species of Diapoma, one from the Negro River, a tributary of the Uruguay River in Brazil and Uruguay, and one from the Iguaçu River, in Brazil and Argentina. The new species from the Negro River basin is distinguished from its congeners by the following combination of characters: a black narrow and conspicuous line restricted to the body horizontal septum, incomplete lateral line, tricuspid teeth in the inner series of the premaxilla, and a lower body depth at vertical through the dorsal-fin origin (29.3-32.8% SL in males and 27.7-33.3% SL in females). The new species from the Iguaçu River basin is distinguished from its congeners by the following combination of characters: a discontinuous lateral line, adipose fin hyaline, longer anal-fin base (26.5-32.4% SL), and a longitudinal black stripe along the median region of caudal-fin rays. Additionally, we updated the molecular phylogeny of the genus, including new sequences from these two new species and Diapoma thauma. An identification key for species of Diapoma is presented, modified from previous study.(AU)

Neste trabalho descrevemos duas espécies novas de Diapoma, uma do rio Negro, afluente do rio Uruguai, Brasil e Uruguai, e a segunda do rio Iguaçu, Brasil e Argentina. A espécie nova do rio Negro é diagnosticada de suas congêneres pela combinação das seguintes características: uma linha preta, estreita e conspícua restrita ao septo horizontal do corpo, linha lateral incompleta, dentes da série interna da pré-maxila tricuspidados, e baixa altura do corpo na vertical que passa pela origem da nadadeira dorsal (29,3-32,8% SL em machos e 27,7-33,3% SL em fêmeas). A espécie nova do rio Iguaçu é diagnosticada de suas congêneres pela combinação das seguintes características: linha lateral descontínua, nadadeira adiposa não pigmentada de preto, base da nadadeira anal longa (26,5-32,4% SL) e raios médios da nadadeira caudal com uma linha preta longitudinal. Adicionalmente, atualizamos a filogenia molecular do gênero, incluindo novas sequências destas duas espécies novas e de Diapoma thauma. Uma chave de identificação para as espécies de Diapoma é apresentada, modificada de estudo anterior.(AU)

Fístula retroauricular complicada asociada a disgenesia mayor del oído externo. Reporte de un caso / Complicated retroauricular fistula with external ear major dysgenesis. A case report

Las anomalías de las hendiduras y arcos branquiales son la segunda causa más común de lesiones congénitas de cabeza y cuello en niños. Representan el 8% de todas las malformaciones congénitas y el 30% de las de cabeza y cuello. Pueden permanecer asintomáticas o manifestarse como una tumefacción y asociarse a infecciones recurrentes supuradas en la región preauricular, subauricular o retroauricular, en las regiones de la parótida y/o del cuello. Fueron reportados casos asociados a malformaciones óticas.La tomografía computada de alta resolución y la resonancia magnética con gadolinio pueden ayudar al diagnóstico. El tratamiento médico antibiótico está indicado ante una infección aguda, además de incisión y drenaje en los procesos abscedados. El tratamiento definitivo es quirúrgico con la extirpación completa de la lesión. Se presenta a un niño de 6 años de edad con infección aguda en la región retroauricular derecha en el oído disgenésico

Branchial cleft anomalies are the second most common congenital head and neck lesions in children. The first congenital head and neck lesion is thyroglossal duct cysts. First branchial cleft anomalies are rare congenital head and neck malformations (8% of branchial cleft anomalies).The initial clinical manifestation was recurrent infections and/or discharge in auricular, periauricular, parotid or upper neck regions. These anomalies are extremely rare and other associated facial malformations were described.The computed tomography and magnetic resonance can help the diagnosis. The antibiotic treatment is indicated in acute infection. Also, incision and drainage are recommended in abscessed processes.In this report, we present a case of retroauricular sinus infection in a 6-year-old child with congenital ear anomalies

The effects of altered BMP4 signaling in first branchial-arch-derived murine embryonic orofacial tissues / 国际口腔科学杂志·英文版

The first branchial arch (BA1), which is derived from cranial neural crest (CNC) cells, gives rise to various orofacial tissues. Cre mice are widely used for the determination of CNC and exploration of gene functions in orofacial development. However, there is a lack of Cre mice specifically marked BA1's cells. Pax2-Cre allele was previously generated and has been widely used in the field of inner ear development. Here, by compounding Pax2-Cre and R26R-mTmG mice, we found a specific expression pattern of Pax2

Mini-incision with endoscope-assisted surgery for bilateral congenital second branchial cleft fistula and a pedigree report / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the feasibility and efficacy of mini-incision with endoscope-assisted resection for bilateral congenital second branchial cleft fistula, and to report on a rare pedigree. Methods: The clinical data of 5 patients with bilateral congenital second branchial fistula admitted in Fujian Provincial Hospital from April 2007 to December 2018 were retrospectively reviewed, including 2 males and 3 females, aged from 3 to 31 years old. The surgical strateges and clinical experience of single mini-incision with endoscope-assisted fistulectomy were summarized, and a rare pedigree was reported. Results: In five patients, Case 1 to Case 4 were treated with bilateral endoscopic-assisted fistula high ligation with titanium clips and removal through a single small incision under general anesthesia. No obvious complications occurred after the operation. The patients were followed up for 40-164 months with no fistula recurrence. Case 5 gave up surgical resection and was followed up for 24 months with acute infection attack once. Case 2 and Case 4 came from the same family. In this family, 7 out of 31 members of four generations had second branchial cleft fistulas, of which 4 were bilateral and 3 were right. Pedigree analysis was consistent with autosomal dominant inheritance. No deafness, preauricular tag, external and middle ear deformity and kidney malformation were found in the family members. Conclusions: Bilateral congenital second branchial cleft fistula is rare. Surgical resection is the preferred treatment. Mini-incision with endoscopic-assisted fistula high ligation with titanium clip and resection has clear operative field, ideal cosmetic effect and definite curative effect.

Parasitic lesions in fish in the Federal District, Brazil

Brazil has a vast territory and favorable climatic conditions that allow the cultivation of freshwater fish. The intensification of the productive system can cause an imbalance in the aquatic environment as a result of poor water quality, nutritional deficiencies and infectious or parasitic diseases. The laboratory diagnosis and the determination of the prevalence of the main lesions, which occur in a certain region, help to guide towards the etiological diagnosis. This study aimed to describe the main parasitic lesions in fish in the routine at the Veterinary Pathology Laboratory of the Universidade de Brasília (UnB). All records of fish with parasitic lesions were recovered. Those cases in which there was an intralesional parasite and which presented lesions compatible with the parasite were included. The screening of ectoparasites was done by scraping the superficial mucus from the gills and skin. Organ sections were routinely processed for histopathologyand stained with hematoxylin-eosin (HE). In some records, parasitological identification was carried. The information was divided into the species of the affected fish, epidemiology of the outbreak (water quality, temperature, type of breeding), lesion distribution, etiology and macroscopic and microscopic changes. The resulting data was organized in absolute frequency and percentage. In this study, 22 cases were counted, between individual deaths and outbreaks, totaling 83 necropsied teleost fish. Inflammatory changes of parasitic origin were seen in 13/22 (59%) of the cases had lesions of parasitic origin. Skin lesions and gills were the most relevant. Macroscopically, red areas or spots of hyperemia or hemorrhage on the body surface were the most prevalent findings. Under microscopy, proliferative gill inflammation was the most relevant diagnosis. Pscinoodinium pilullare (Dinoflagellida), Ichthyophthirius multifiliis (Ciliophora), and monogenetic worms (Monogenea) were the main parasites found. Trichodina sp. (Ciliophora), Ichtyobodo sp. (Kinetoplastida), Amoebas, and Chilodonella sp. (Ciliophora), were seen in fewer numbers. An unusual case suggestive of parasitism by Eustrongylides sp. (Nematoda), in a pirá-brasília (Simpsonichthys boitonei), specimen has been recorded. The diagnoses were based on epidemiology, anatomopathological and parasitological findings. The most frequent and significantly lethal lesion in the study was proliferative and / or hyperplastic branchitis. Proliferative branchitis with lamellar epithelial hyperplasia (LEH) is a response to some type of chemical or mechanical injury to the gill epithelium in order to protect the capillaries from further damage or microbial penetration. However, it also increases the diffusion distance between capillaries and the environment and, therefore, hinders breathing, excretory and osmoregulatory functions. Protozoan infections and monogenetic worms in general generated LEH and skin lesions of mechanical origin. Secondary bacterial infection, were observed in this parasitosis determining the cause of death of the fish. Its pathogenicity comes from the lesions caused by the colonization and histophagy of the epithelial surfaces, mainly gills and skin, causing epithelial proliferation, lamellar cell fusion, epithelial cell degeneration and necrosis forming several ulcers in the epithelium after the release of mature trophies. The pathogenesis of parasitism by Eustrongylides spp. is considerable when there is a large quantity of these larvae that can cause intestinal obstruction, rupture and compression of viscera, of greater importance in small fish. The main parasites of necropsied fish were protozoa and monogenetic worms, which mainly cause branquitis and dermatitis in varying grades.(AU)

No todo son adenopatías / Not everything is an adenopathy

Las tumoraciones cervicales son habituales en niños. Lo más frecuente es que se trate de adenopatías secundarias a infecciones, virales o bacterianas. No obstante, es conveniente tener en cuenta otras causas de adenopatías y la existencia de otros procesos que pueden confundirse con ellas. La exploración mediante ecografía es la técnica de elección para diferenciar la lesión y determinar sus características. Presentamos el caso clínico de una paciente con adenopatías cervicales, a la que se diagnosticó una masa congénita con ecografía en Atención Primaria

Cervical masses are frequent in children. Most often they are lymphadenopathies secondaries to infection, viral or bacterial. However, we should think in other causes of adenopathy and other processes that may be confused with them. Ultrasound examination is the technique of choice to differentiate the lesion and determine its characteristics. We present a patient with cervical adenopathies, who was diagnosed a congenital mass with ultrasound in Primary Care

Branchial cleft cyst in the parotid gland in a human immunodeficiency virus-negative patient

In branchial lymphoepithelial cyst (BLEC), which is also known as branchial cleft cyst, the remnants of a branchial arch develop into a cyst, causing swelling. The first case of BLEC in the parotid gland was reported by Hildebrant in 1895. Since then, BLEC in the parotid gland has continued to be reported, but in rare cases. A 45-year-old man presented to our hospital with a swelling of the left cheek of approximately 6 months’ duration. The patient underwent a superficial parotidectomy and was pathologically diagnosed with BLEC. Of note, this was the first case of non-human immunodeficiency virus (HIV)-related BLEC of the parotid gland in South Korea. BLEC is a benign condition, but its treatment depends on the presence of HIV infection. In HIV-negative patients, BLEC does not require a further work-up to evaluate metastasis. Our case report describes the diagnosis and treatment of BLEC in a patient without HIV.

Hey1 gene expression patterns during the development of branchial arches and facial prominences / Patrones de expresión del gen Hey1 durante el desarrollo de arcos branquiales y prominencias faciales

ABSTRACT Objective. The present study aimed to describe in detail the expression patterns of the gene Hey1, an effector of the Notch pathway, during the development of branchial arches and facial prominences. Materials and methods. Fertilized chicken (Gallus gallus) eggs obtained from a local egg farm were incubated at 37.5 -38.5ºC with 70% relative humidity until the embryos reached Hamilton-Hamburger stages HH14 through HH25. Digoxigenin-UTP labeled probes Hey1 were generated from linearized plasmids with either T3 polimerase for in vitro transcription. Whole-mount in situ hybridization was then performed. At least 3 replicates (n=3) were obtained for each stage. To confirm the results observed in whole embryos, sagittal and coronal cryosectioning was performed using a thickness of 10 µm. Results. During developmental stages HH14 and HH18, Hey1 gene expression was localized to the endoderm of branchial pouches. Hey1 gene expression was also observed in the epithelium that covers the maxillary and mandibular prominences during developmental stages HH19 and HH21, as well as in the nasal epithelium between HH19 and HH25. Transcripts were also detected in the epithelium that covers the frontonasal prominence during stage HH21. Conclusions. These expression patterns suggest the participation of this component of the Notch signaling pathway in craniofacial morphogenesis, possibly establishing pharyngeal segmentation patterns during early stages and/or regulating cell proliferation and differentiation during the late stages of facial development.

RESUMEN Objetivo. El presente estudio tuvo como objetivo describir detalladamente los patrones de expresión del gen Hey1, un efector de la vía Notch durante el desarrollo de arcos branquiales y prominencias faciales. Materiales y métodos. Se incubaron huevos fertilizados de pollo (Gallus gallus) obtenidos de una granja local entre 37.5-38.5ºC con humedad relativa del 70% hasta que los embriones alcanzaron los estadios HH14 hasta HH25 de Hamilton-Hamburger. Las sondas Hey1 marcadas con digoxigenina-UTP se generaron a partir de plásmidos linearizados con T3 polimerasa por transcripción in vitro. Luego se realizó hibridación in situ sobre embriones completos. Se obtuvieron al menos 3 repeticiones (n=3) para cada estadio. Para confirmar los resultados observados en embriones completos, se realizaron cortes sagitales y coronales de 10 µm. Resultados. Durante los estadios de desarrollo HH14 y HH18, la expresión del gen Hey1 se localizó en el endodermo de las bolsas branquiales. La expresión génica de Hey1 también se observó en el epitelio que cubre las prominencias maxilares y mandibulares durante las etapas de desarrollo HH19 y HH21, así como en el epitelio nasal entre HH19 y HH25. También se detectaron transcritos de Hey1 en el epitelio que cubre la prominencia frontonasal durante la etapa HH21. Conclusiones. Estos patrones de expresión sugieren la participación de este componente de la vía de señalización Notch en la morfogénesis craneofacial, posiblemente estableciendo patrones de segmentación faríngea durante las primeras etapas y / o regulando la proliferación y diferenciación celular durante las últimas etapas del desarrollo facial.