Short Arm Research Articles

1p36 deletion syndrome: Review and mapping with further characterization of the phenotype, a new cohort of 86 patients.

Chromosome 1p36 deletion syndrome (1p36DS) is one of the most common terminal deletion syndromes (incidence between 1/5000 and 1/10,000 live births in the American population), due to a heterozygous deletion of part of the short arm of chromosome 1. The 1p36DS is characterized by typical craniofacial features, developmental delay/intellectual disability, hypotonia, epilepsy, cardiomyopathy/congenital heart defect, brain abnormalities, hearing loss, eyes/vision problem, and short stature. The aim of our study was to (1) evaluate the incidence of the 1p36DS in the French population compared to 22q11.2 deletion syndrome and trisomy 21; (2) review the postnatal phenotype related to microarray data, compared to previously publish prenatal data. Thanks to a collaboration with the ACLF (Association des Cytogénéticiens de Langue Française), we have collected data of 86 patients constituting, to the best of our knowledge, the second-largest cohort of 1p36DS patients in the literature. We estimated an average of at least 10 cases per year in France. 1p36DS seems to be much less frequent than 22q11.2 deletion syndrome and trisomy 21. Patients presented mainly dysmorphism, microcephaly, developmental delay/intellectual disability, hypotonia, epilepsy, brain malformations, behavioral disorders, cardiomyopathy, or cardiovascular malformations and, pre and/or postnatal growth retardation. Cardiac abnormalities, brain malformations, and epilepsy were more frequent in distal deletions, whereas microcephaly was more common in proximal deletions. Mapping and genotype-phenotype correlation allowed us to identify four critical regions responsible for intellectual disability. This study highlights some phenotypic variability, according to the deletion position, and helps to refine the phenotype of 1p36DS, allowing improved management and follow-up of patients.

A multiplex PCR-based sensitive and specific method for detecting Y chromosome material in patients with Turner syndrome

To develop a multiplex PCR method for a rapid detection of Y chromosome-specific sequences in patients with Turner syndrome. Nine genes were selected from various regions of the Y chromosome for designing the primers, which included SRY, TBL1Y, TSPY on the short arm of the Y chromosome, DDX3Y, HSFY1, RPS4Y2 and CDY1 on the long arm of Y chromosome and SHOX in the short arm and SPRY3 in the long arm of the pseudoautosomal region (PAR) of X and Y chromosomes. A multiplex PCR method for the nine genes in Y chromosome was established and optimized. The sensitivity was tested by using different amounts of genomic DNA. A total of 36 patients with Turner syndrome and a patient with male dwarfism with karyotype of 46, X, +mar were examined by the multiplex PCR method for the existence of materials from the Y chromosome. The optimization results of the multiplex PCR reaction system (50 μL) showed that when the final concentration of upstream and downstream of each pair of primers was 0.1 μM, the multiplex PCR reaction of the 9 pairs of primers clearly amplified the target with the expected band size, and there was no non-specific amplification. The bands were clearly visible when the amount of genomic DNA in the multiple PCR reaction system was as low as 1 ng. By using the method, we have examined the 36 patients with Turner syndrome. One patient with Turner syndrome with karyotype of 45,X[40]/47XYY[21] amplified specific seven genes on Y chromosome, 35 patients with Turner syndrome amplified only two target genes SHOX and SPRY3, but not the other seven specific genes on the Y chromosome, which was in keeping with the clinical manifestations of such patients. This study established a multiplex PCR reaction system with nine genes, which can quickly and accurately screen Y chromosome materials in patients with Turner syndrome. It has the advantages of low cost, simple operation, high specificity and rapid turn-around time, and can be used to detect Turner syndrome patients with Y chromosome material in time. The method has provided a diagnostic basis for preventive gonad resection to prevent malignant gonadal tumors.

Interstitial Duplication on Chromosome 3p14.3p13 in an Adolescent with Dysmorphic Features and Autism, Case Report

Abstract Introduction/Objective The genetic etiology of autism spectrum disorders is only partially understood. Here we describe a 16-year-old male diagnosed with autism at two years of age. He has dysmorphic features, severe cognitive disability, and history of cryptorchidism. A review of systems was significant for slightly coarse features overall, with deep-set eyes, pinched nasal bridge with wide nasal tip, and widely spaced teeth. A large pectus excavatum deformity was also noted. Although the parent’s genetic testing concluded that this likely represents de novo mutation, it is worth mentioning that our patient has a 13-year-old female maternal first cousin with learning difficulties that were said to be less severe. There is no other family history of autism. Methods/Case Report Chromosome analysis showed an abnormal karyotype identifying a duplication of the short arm of chromosome 3 from p13 to p14.3, 46,XY,dup(3)(p13p14.3). Additional microarray testing confirmed this duplication and defined the size as 14.9 Mb. Regions of homozygosity of 29.8 Mb were also identified, representing about 1% of the autosomal genome. The duplicated area includes over 70 genes, thirteen of which are known Mendelian disease genes (IL17RD, HESX1, APPL1, FLNB, DNASEIL3, PDHB, ACOX2, ATXN7, SLC25A26, EOGT, LMOD3, MITF, and FOXP1). Duplications of 3p are extremely rare and can be de novo or inherited from a parent with a balanced translocation. Individuals with these better-described chromosome 3p duplications typically present with intellectual and developmental disabilities, such as autism, as 3p duplications typically present with intellectual and developmental disabilities, such as autism and distinctive dysmorphic features. This duplication has never been reported as a known syndrome and has minimal overlap with copy number variants (CNVs) among healthy individuals. Results (if a Case Study enter NA) NA Conclusion Autism spectrum disorders are largely characterized by speech, communication, and social impairment of varying degrees. Diagnosis is typically made on clinical grounds, but in 14-35% of cases, a genetic basis for the disorder, typically either due to a single gene disorder or a chromosomal deletion/duplication, may be found. This case report describes a rare finding of a 14.9 Mb interstitial duplication on chromosome 3p.

Development of diagnostic SNP markers for identification of rye 1RS translocations in wheat

AbstractWheat (Triticum aestivum L.) wild relatives are rich genetic resources for genetic improvement of wheat. The short arm of rye (Secale cereale L.) chromosome 1 (1RS) contains many beneficial genes for resistance to drought, insects, and diseases, therefore the 1RS wheat translocations have been widely used in wheat breeding programs worldwide. To facilitate marker‐assisted identification of 1RS in wheat breeding, we designed a set of Kompetitive allele‐specific polymerase chain reaction (KASP) markers to detect the translocations of 1RS with long arm from wheat chromosome 1A (1RS.1AL) and 1RS with long arm from wheat chromosome 1B (1RS.1BL) based on the sequence flanking single nucleotide polymorphisms (SNPs) generated by genotyping‐by‐sequencing (GBS) between wheat and rye panels. We identified a set of three SNP markers for diagnosis and differentiation of the two 1RS translocations in wheat. Among the three KASP markers, 1AS‐05056 is 1RS specific, and clearly separated the genotypes with 1RS (1RS.1AL and 1RS.1BL) from those without the two 1RS translocations; whereas 1AS‐29875 clearly identified the 1RS.1AL translocation and 1BS‐16654 separated 1RS.1BL from 1RS.1AL. The three new KASP markers together accurately identified the presence of 1RS translocation on either 1A or 1B chromosome in a panel of 95 representative U.S. winter wheat accessions, which were further validated in two additional U.S. hard winter wheat panels. Therefore, they can be used for routine screening of 1RS.1BL and 1RS.1AL translocation lines in wheat breeding programs.

Validation of a QTL on Chromosome 1DS Showing a Major Effect on Salt Tolerance in Winter Wheat.

Salt stress is one the most destructive abiotic stressors, causing yield losses in wheat worldwide. A prerequisite for improving salt tolerance is the identification of traits for screening genotypes and uncovering causative genes. Two populations of F3 lines developed from crosses between sensitive and tolerant parents were tested for salt tolerance at the seedling stage. Based on their response, the offspring were classified as salt sensitive and tolerant. Under saline conditions, tolerant genotypes showed lower Na+ and proline content but higher K+, higher chlorophyll content, higher K+/Na+ ratio, higher PSII activity levels, and higher photochemical efficiency, and were selected for further molecular analysis. Five stress responsive QTL identified in a previous study were validated in the populations. A QTL on the short arm of chromosome 1D showed large allelic effects in several salt tolerant related traits. An expression analysis of associated candidate genes showed that TraesCS1D02G052200 and TraesCS5B02G368800 had the highest expression in most tissues. Furthermore, qRT-PCR expression analysis revealed that ZIP-7 had higher differential expressions under saline conditions compared to KefC, AtABC8 and 6-SFT. This study provides information on the genetic and molecular basis of salt tolerance that could be useful in development of salt-tolerant wheat varieties.

PO29 Presentation Time: 7:40 AM: Dosimetric Review to Organs at Risk from Short Course Adjuvant Vaginal Cuff Brachytherapy in Early Endometrial Cancer Compared to Standard of Care (SAVE)

<h3>Purpose</h3> To evaluate dosimetric characteristics to organs at risk (OARs) from short course adjuvant vaginal cuff brachytherapy (VCB) in early endometrial cancer compared to standard of care in a multi-institutional prospective randomized trial. <h3>Materials and Methods</h3> 108 patients requiring VCB were randomized to an experimental short course arm (11 Gy x 2Fx to surface) and standard of care arms: 7 Gy x 3Fx to 5mm, 5-5.5 Gy x 4Fx to 5mm, and 6 Gy x 5Fx to surface. Rectum, bladder, sigmoid, small bowel, and urethra were contoured retrospectively on the planning CT and doses to OARs were compared by arm. Absolute doses for each OAR (D2cc and D0.1cc), and from each fractionation scheme, were converted to 2 Gy equivalent dose (EQD2; α/β=3 Gy). Each standard of care arm was compared to the experimental arm separately using a one-way analysis of variance, followed by pairwise comparisons using Tukey's HSD test. Additionally, treatment planning system reported point doses (mid cylinder) to the vaginal mucosa was converted to EQD2 (α/β=10 Gy) and evaluated. <h3>Results</h3> The experimental arm had significantly lower doses for rectum, bladder, sigmoid, and urethra when compared to the 7 Gy x3 and 5-5.5 Gy x4 fractionation schemes; however, the experimental arm did not differ statistically from the 6 Gy x5 fractionation scheme. For small bowel doses, none of the standard of care fractionation schemes were statistically different than the experimental arm (see fig. 1f). The highest D2cc EQD2 doses to the examined critical structures were observed to come from the most common dose fractionation scheme of 7 Gy x 3Fx (see fig. 1a-e). The data revealed similar results for the D0.1cc EQD2 doses for the urethra. Furthermore, average reported EQD2 doses to the vaginal mucosa were 40.32 Gy (σ=1.97 Gy) for 11 Gy x2, 56.53 Gy (σ=6.29 Gy) for 7 Gy x3, 47.81 Gy (σ=2.97 Gy) for 5-5.5 Gy x 4, and 41.69 Gy (σ=1.29 Gy) for 6 Gy x 5. With a short median follow up of one year, there have been no isolated vaginal recurrences. <h3>Conclusions</h3> Doses to OARs were generally low; however, in the highest quintile significant doses can be delivered with VCB. Experimental short course VCB of 11 Gy x 2Fx to the surface provides a comparable biologic effective dose to standard of care courses. Experimental short course VCB was found to reduce or be comparable to D2cc and D0.1cc EQD2 doses to rectum, bladder, sigmoid, small bowel, and urethra critical structures. This may translate into a comparable or lower rate of acute and late side effects.

Short axis upper arm showing the ulnar nerve with the humerus below.

Short axis upper arm showing the ulnar nerve with the humerus below.

PMON270 Coexistence of 46, XX Testicular Disorder of Sex Development and 11β-Hydroxylase Deficiency: In Vivo and In Vitro Studies

Abstract Background 46, XX testicular disorder of sex development and 11β-hydroxylase deficiency (11β-OHD) are two extremely rare types of disorder of sex development (DSD). No coexistence has been reported yet. Clinical Case A 22-year-old male showed small testes (volume=4ml) and gynecomastia for 2 years. At the age of 3.5, he was diagnosed as simple virilizing type of 21-hydroxylase deficiency (21-OHD) for the manifestations of the enlarged penis (length=7.5cm), hyperpigmentation, and normal blood pressure. Adrenal ultrasound suggested bilateral adrenal hyperplasia (both length * width = 2.4*0.9cm) and X-ray showed accelerated bone age (12.5 years old). Irregular cortisone acetate 6.25mg per day was administrated since then. At the age of 18, he was sent to emergency for aortic aneurysm rupture and heart failure, and hypertension urgency was diagnosed. Sanger sequencing revealed that he carried compound heterozygous variants in CYP11B1 gene: NM_000497.3: c.905_907delinsTT and NM_000497.3: c.954+7C&amp;gt;T, which were respectively inherited from his father and mother. Pathogenesis of the novel intron variant (c.954+7C&amp;gt;T) was further verified in COS7, CHO, and 273T cell lines by transfecting wild-type and mutant-type minigene, extracting RNA of cells, reversing RNA into cDNA, PCR, and Sanger sequencing. The result showed that the c.954+7C&amp;gt;T variant introduced a cryptic donor splice site at a site 5 bp downstream of the 3' natural splice donor site of exon 5. The level of adrenal steroids detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was consistent with the characteristics of 11β-OHD. The patient was treated by neoplasty of thoracic aortic aneurysm and added anti-hypertension drugs in addition to glucocorticoid after surgery. Unexpectedly, the abnormalities of male characteristics including small testes and gynecomastia appeared since adolescence. Laboratory tests showed an elevated level of FSH and LH and azoospermia. Scrotum Ultrasound showed bilateral testicular dysplasia (right: 1.9*1.4*0.8cm, left: 1.9*1.3*0.8cm). To figure out the unexplainable results, whole-exome sequencing was performed and revealed two copy number variants: the duplication of Xp22.33-q28 which spans 151.32Mb, and the deletion of Yp11.2-q11.23 which spans 23.34Mb. Meanwhile, the karyotype test by culturing blood lymphocytes showed 46, XX (100/100), which confirmed the diagnosis of 46, XX testicular DSD. Fluorescence in situ hybridization (FISH) revealed the presence of the SRY gene, which is translocated into the short arm of the X chromosome. Testosterone replacement is unnecessary for the normal level of testosterone (T) (4.44ng/ml, N: 1.75-7.81ng/ml), which is the result of the combined action of 11β-OHD with adrenal T accumulation and 46, XX testicular DSD with testicular T deficiency. Assisted reproduction consulting should start early. Conclusion This is the first case demonstrating the possibility of coexistence of 46, XX testicular DSD and 11β-OHD and hinting at the significance of a combination of clinical and molecular diagnostic methods. Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

OR04-4 Loss of KDM1A in Bilateral Macronodular Adrenal Hyperplasia With GIP-Dependent Cushing's Syndrome and in Acromegaly With Paradoxical GH Response to Oral Glucose

Abstract Context Primary bilateral macronodular adrenal hyperplasia (PBMAH) with glucose-dependent insulinotropic polypeptide (GIP)-dependent Cushing's syndrome is caused by ectopic expression of GIP receptor (GIPR) in the adrenal lesions. Such ectopic expression of GIPR was also reported in other endocrine neoplasm, notably in somatotroph pituitary adenomas from acromegalic patients with paradoxical increase of GH after oral glucose load, suggesting a common molecular pathogenesis. We aimed to identify the driver event responsible for GIP-dependent PBMAH with Cushing's syndrome and ectopic GIPR expression in somatotropinomas. Methods We conducted an international, multicenter, cohort study. We collected blood and adrenal samples from patients who had undergone unilateral or bilateral adrenalectomy for GIP-dependent PBMAH with Cushing's syndrome. Adrenal samples from patients with PBMAH and Cushing's syndrome without food-dependent cortisol production were used as controls. We further collected somatotropinoma specimens from acromegaly patients followed at two expert endocrine centers in France. Results 17 patients with familial or sporadic GIP-dependent PBMAH with Cushing's syndrome were studied. We identified germline heterozygous mutations in the lysine demethylase 1A (KDM1A) gene in all 17 patients. We further identified a recurrent deletion of the short arm of chromosome 1 harboring the KDM1A locus in the adrenal lesions of affected patients. None of the 25 patients in the control group had KDM1A germline or somatic alterations. Concomitant genetic inactivation of both KDM1A alleles resulted in loss of KDM1A expression in the adrenal lesions. RNA-sequencing revealed the global impact of KDM1A loss in adrenal tissue on gene transcription and identified differentially regulated genes including those encoding for GIPR and other G-Protein-Coupled Receptors that may be involved in adrenal tumorigenesis and regulation of steroidogenesis. In vitro pharmacologic inhibition, silencing and knock-out by CRISPR-Cas9 genome editing of KDM1A led to an increase in GIPR transcripts and protein in human adrenocortical H295R cells. Somatotropinoma samples from 78 patients with acromegaly were studied. 24% of these patients presented with a paradoxical rise of GH after oral glucose load and expressed ectopically GIP-receptor in their somatotropinoma. None of the somatotropinomas harbored KDM1A pathogenic variants, but those from patients with paradoxical GH response displayed a recurrent chromosome 1p loss. Discussion We identified germline inactivating KDM1A mutations and loss of heterozygosity as a genetic predisposition to GIP-dependent PBMAH with Cushing's syndrome following a tumor suppressor gene model of tumorigenesis. We currently perform genetic screening in first-degree relatives of patients with GIP-dependent PBMAH with Cushing's syndrome and clinical examination with biochemical testing in asymptomatic KDM1A variant carriers. We did not identify somatic KDM1A mutations in somatotropinomas expressing GIPR ectopically, however their recurrent 1p chromosome loss suggests that KDM1A haploinsufficiency may contribute to GIPR expression in those tumors. Presentation: Saturday, June 11, 2022 12:15 p.m. - 12:30 p.m.

The Role of Eif2s3y in Mouse Spermatogenesis and ESC.

Eukaryotic translation initiation factor 2 subunit 3 and structural gene Y-linked (Eif2s3y) gene, the gene encoding eIF2γ protein, is located on the mouse Y chromosome short arm. The Eif2s3y gene is globally expressed in all tissues and plays an important role in regulating global and gene-specific mRNA translation initiation. During the process of protein translation initiation, Eif2s3x (its homolog) and Eif2s3y encoded eIF2γ perform similar functions. However, it has been noticed that Eif2s3y plays a crucial role in spermatogenesis, including spermatogonia mitosis, meiosis, and spermiogenesis of spermatids, which may account for infertility. In the period of spermatogenesis, the role of Eif2s3x and Eif2s3y are not equivalent. Importance of Eif2s3y has been observed in ESC and implicated in several aspects, including the pluripotency state and the proliferation rate. Here, we discuss the functional significance of Eif2s3y in mouse spermatogenesis and self-renewal of ESCs.

45,X male – rare case of unbalanced translocation of Y chromosome to chromosome 2 presenting with developmental delay, learning difficulty and obesity

A male phenotype accompanied by a 45,X karyotype is rare. It may occur due to Y chromosomal translocation or insertion to X/autosome. Clinical presentation may vary depending on the presence of the Y chromosomal locus and the degree of loss of autosome material. 45,X males can present with short stature and Turner syndrome phenotype due to haploinsufficiency of genes which are normally expressed in both X and Y chromosomes. The presence of the sex-determining region Y (SRY) gene leads to the differentiation of bipotential gonads to testis. Most individuals go through puberty normally, but some may need pubertal induction for delayed puberty. Rarely some can have a pubertal arrest. The risk of gonadoblastoma is minimal in these individuals due to functioning testicular tissue. The azoospermia factor (AZF) region is found on the long arm of the Yq chromosome and is needed for spermatogenesis. In a 45,X male with unbalanced translocation of Y chromosome, spermatogenesis can be affected due to the lack of AZF leading to Sertoli cell-only syndrome. This will have an implication on fertility in adult life. We present a 14-year-old boy with developmental delay, learning difficulties and subtle dysmorphic features who was diagnosed with 45,X,der(2)t(Y:2)(?:p25). Fluorescence in situ hybridisation analysis revealed translocation of SRY (Yp11.3) to the terminal part of the short arm of chromosome 2 resulting in the deletion of most of the Y chromosome (Yp11.2-q12) and part of chromosome 2(2p25.3). This is the first case where SRY translocation to chromosome 2 presents with the above clinical presentation. 45,X karyotype is rare in male. It may occur due to SRY translocation or an insertion to X/autosome. SRY gene translocation to chromosome 2 has been not reported in the literature. Clinical presentation can be varied due to degree of loss of chromosomal material. Due to loss of AZF region found on the long arm of the Yq, spermatogenesis can be affected. Loss of 2p25 leads to learning difficulty and obesity.

105. Clinical whole-genome sequencing identifies NSD3 as the correct fusion partner of NUP98 in a patient with acute myeloid

<h3>Background</h3> Only rare cases of acute myeloid leukemia (AML) have been shown to harbor a t(8;11)(p11.2;p15.4). This translocation is believed to involve the fusion of NSD3 or FGFR1 with NUP98; however, apart from targeted mRNA quantitative PCR analysis, no molecular approaches have been utilized to define the chimeric fusions present in these rare cases. <h3>Case Presentation</h3> Here we present the case of a 51-year-old female with AML with myelodysplastic-related morphologic changes, 13q deletion and t(8;11), where initial fluorescence in situ hybridization (FISH) assays were consistent with the presence of NUP98 and FGFR1 rearrangements, and suggestive of NUP98/FGFR1 fusion. Using a streamlined clinical whole-genome sequencing approach, we resolved the breakpoints of this translocation to intron 4 of NSD3 and intron 12 of NUP98, indicating NUP98/NSD3 rearrangement as the underlying aberration. Furthermore, our approach identified small variants in WT1 and STAG2, as well as an interstitial deletion on the short arm of chromosome 12, which were cryptic in G-banded chromosomes. <h3>Conclusions</h3> NUP98 fusions in acute leukemia are predictive of poor prognosis. The associated fusion partner and the presence of co-occurring mutations, such as WT1, further refine this prognosis with potential clinical implications. Using a clinical whole-genome sequencing analysis, we resolved t(8;11) breakpoints to NSD3 and NUP98, ruling out the involvement of FGFR1 suggested by FISH while also identifying multiple chromosomal and sequence level aberrations.

Noninvasive Technique to Monitor the Pressure under a Cast: A Mobile Application-Friendly Bluetooth Pressure Sensor.

The purpose of this study was to design a sensor that could monitor the skin-cast contact surface pressure (SCCSP) of a limb under a cast and inform the user via a mobile application when the pressure increases. In this experimental study, an infant sphygmomanometer cuff was initially placed on the forearm of 10 volunteers. A pressure sensor with a Bluetooth chip was then placed on the volar aspect of the forearm. Short arm plaster was applied with synthetic cast material. The SCCSP under the plaster was measured by the sensor and the measured values were transmitted to a mobile application via a Bluetooth chip. The mobile application processed the data from the chip and converted it to mmHg. Intracompartmental pressure (ICP) values were categorized as 0, 10, 20, 30, 40, 50, 60, and 75 mmHg. The highest SCCSP was 75 mmHg CP, while the lowest was 0 mmHg CP. The correlation coefficient of the mean pressure values was 0.993 (p ≤ 0.001) (SD 0.002, range 0.989-0.997), and there was a significant relationship between ICP and SCCSP values (p ≤ 0.05). We can monitor SCCSP, detect limb swelling, and notify the user via a mobile application by using Bluetooth pressure sensors.

Comparative cytogenetic within Achillea genus (Asteraceae) in Iran

The cytological features including chromosome numbers and detailed chromosome measurements of 46 populations of ten Achillea species (A. aleppica, A. bibersteinii, A. eriophora, A. filipendula, A. millefolium, A. nobilis, A. pachycephala, A. tenuifolia, A. vermicularis, and A. wilhelmsii) were reported from Iran. Sample plants and seeds were collected from diff erent geographical regions of Iran. Total chromosome length, long/ short arm ratio, centromere index, total form percentage, inter and intra-chromosomal index, different relative lengths of the mitotic chromosomes were calculated. The populations showed one basic chromosome number x = 9 with two ploidy levels (2x and 4x). The chromosome numbers of A. filipendula, A. nobilis and A. pachycephala, for the first time are new to science and new ploidy levels are reported for A. biebersteinii (4x) and A. wilhelmsii (2x). The mean of chromosome length ranged from 3.8 μm in A. millefolium from Semnan, Chasham, Sirat to 6.37 μm in A. nobilis from Golestan, Agh Emam. Karyotype analysis indicated that chromosomes of Achillea taxa have predominantly median and sub-median centromeres. Several karyological aspects of these taxa are discussed in the light of the cytogenetic data.

Establishment and Characterisation of Human Clear Cell Sarcoma and Malignant Melanoma Cell Lines.

Clear cell sarcoma of soft tissue (CCSST) and conventional malignant melanoma (MM) are rare and aggressive tumours with similarities in morphology and the expression of melanocytic markers. We established two CCSST cell lines (FU-CCSST-1 and FU-CCSST-2) from soft tissues of the patella and supraclavicular. A MM cell line (FU-MM-1) was established from lymph node metastases of subungual malignant melanoma. FU-CCSST-2 cells were transplantable to immunodeficient mice. Immunohistochemical studies demonstrated tumour cells were negative for cytokeratin AE1/AE3 and positive for S100 protein, HMB45, Melan-A, CD146 and SOX10 in all specimens. FU-CCSST-1 and FU-CCSST-2 harboured t(12;22)(q13;q12) translocations with expression of the EWSR1/ATF1 fusion gene. FU-MM-1 demonstrated loss of the short arm of chromosome 9 and harboured wild-type BRAF (codon 469 and 600) and NRAS (codon 12, 13 and 61). We report the establishment and characterisation of CCSST and MM cell lines that may have utility in the study of pathogenic mechanisms and development of novel therapeutic reagents.

Osteichthyan Fishes from the uppermost Norian (Triassic) of the Fuchsberg near Seinstedt, Lower Saxony (Germany)

Fishes from the uppermost Norian Fuchsberg Quarry near Seinstedt are represented by two taxa that we interpret as a teleosteomorph (complete specimens) and actinistian (scales). Seinstedtia parva gen. et sp. nov. is described; although it was proposed as a possible semionotiform, this study reveals that Seinstedtia possesses a combination of teleosteomorph features, for instance: characteristic pholidophoriform-shaped cranial roof; fusion of skull roof bones; three dorso-posterior infraorbitals, including an enlarged infraorbital 3; one suborbital bone; movable premaxilla; and characteristic-shaped preopercle. In parallel, Seinstedtia possesses a head gently curved anteriorly, with lower jaw protruding slightly in front of upper jaw; supraorbital 1 forming most of antero-dorsal margin of circumorbital ring; one supramaxilla; three extrascapulae; cleithrum with short and broad lower arm; and enlarged clavicle. This character combination places Seinstedtia as a teleosteomorph, family incertae sedis. This fish (total length ca. 50 mm) and some pholidophorids (ca. 70 mm or less; Parapholidophorus nybelini and Pholidoctenus serianus) represent the known smallest teleosteomorphs that inhabited Europe during the Norian. The isolated scales are elasmoid of amioid type ornamented with the elongated ridges of actinistians. This diversity of teleosteomorphs and actinistians in Fuchsberg Quarry during the Triassic indicates a connection to a marine environment.

Integration of genomic analysis and transcript expression of ABCC8 and KCNJ11 in focal form of congenital hyperinsulinism.

The focal form of CHI is caused by an autosomal recessive pathogenic variant affecting the paternal homologue of genes ABCC8 or KCNJ11 and a second somatic event specifically occurring in the affected islet of Langerhans. The approach of this study was to integrate the genetic changes occurring in pancreatic focal lesions of CHI at the genomic and transcriptional level. Patients receiving therapeutic surgery and with proven ABCC8 or KCNJ11 pathogenic variants were selected and analyzed for loss of heterozygosity (LOH), changes in copy number and uniparental disomy (UPD) on the short am of chromosome 11 by molecular microarray analysis and methylation-specific MLPA. Gene expression was analyzed by RT-PCR and Massive Analysis of cDNA Ends (MACE). Both genes, ABCC8 and KCNJ11, are located in proximity to the Beckwith-Wiedemann (BWS) imprinting control region on chromosome 11p15. Somatic paternal uniparental isodisomy (UPD) at chromosome 11p was identified as second genetic event in focal lesions resulting in LOH and monoallelic expression of the mutated ABCC8/KCNJ11 alleles. Of five patients with samples available for microarray analysis, the breakpoints of UPD on chromosome 11p were different. Samples of two patients were analyzed further for changes in gene expression. Profound downregulation of growth suppressing genes CDKN1 and H19 was detected in focal lesions whereas growth promoting gene ASCL2 and pancreatic transcription factors of the endocrine cell lineage were upregulated. Paternal UPD on the short arm of chromosome 11 appears to be the major second genetic event specifically within focal lesions of CHI but no common breakpoint for UDP can be delineated. We show for the first time upregulation of growth promoting ASCL2 (achaete-scute homolog 2) suggestive of a driving factor in postnatal focal expansion in addition to downregulation of growth suppressing genes CDKN1C and H19.

A Systematic Review and Meta-Analysis of Complications among Various Reduction Malarplasty.

Reduction malarplasty is one of the most common aesthetic procedures to improve a wide bizygomatic width and a prominent zygomatic body. Although there are various kinds of modifications, any method is imperfect, while some complications may occur. The purpose of this review was to compare kinds of complications of reduction malarplasty to provide certain suggestions for clinical application. A comprehensive computerized search of scientific literature was performed via the PubMed, Web of Science, and Library of Congress databases, involved in articles from January 1st, 1983 to February 28th, 2022. The outcomes were extracted and analyzed by 3 independent authors, including patient demographics, diagnoses, surgical techniques, postoperative outcomes, and complications. A total of 29 studies covering 6611 patients were included according to the inclusion and exclusion criteria. The L-shaped osteotomy may obtain a better effect when someone has both zygomatic body and arch protrusion. In the view of complications, our conclusion suggested that L-shaped osteotomy without bony resection reduced the zygomatic complex effectively with the lowest incidence of postoperative complications (0.02%). But the amount of bone resection is limited. If increasing bone resection is necessary, L-shaped osteotomy with long arm bony resection and L-shaped osteotomy with short arm bony resection are both preferable choices with lowest incidence of structural and functional complications, respectively. L-shaped osteotomy may obtain a better effect when a patient has both zygomatic body and arch protrusion. L-shaped osteotomy without bony resection reduced the zygomatic complex effectively with the lowest incidence of postoperative complications. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

3D-Printed Short Arm Casts: Reliability, Validity, Feasibility Compared with Conventional Waterproof Fiberglass Casts

Aim: To evaluate the reliability, validity, and feasibility of 3D-printed short arm casts (SACs) versus conventional casts. Methods: Three raters of varying experience applied two conventional and two 3D-printed casts to a participant's dominant wrist to evaluate reliability. Each cast was worn for 24 h and removed the following day after data collection. Results &amp; conclusion: ICCs demonstrated ‘excellent’ intra-rater reliability for clinical effectiveness and patient satisfaction. There were no significant differences between fiberglass and 3D-printed SACs. The 3D-printed SAC group had significantly higher wrist function compared with the fiberglass SAC group. Discussion: This pilot case study demonstrates that 3D-printed short arm casts may be a valid immobilization technique of the wrist compared with conventional waterproof fiberglass casting.

Pathogenesis of Huntington's Disease: An Emphasis on Molecular Pathways and Prevention by Natural Remedies.

Background: Huntington’s disease is an inherited autosomal dominant trait neuro-degenerative disorder caused by changes (mutations) of a gene called huntingtin (htt) that is located on the short arm (p) of chromosome 4, CAG expansion mutation. It is characterized by unusual movements, cognitive and psychiatric disorders. Objective: This review was undertaken to apprehend biological pathways of Huntington’s disease (HD) pathogenesis and its management by nature-derived products. Natural products can be lucrative for the management of HD as it shows protection against HD in pre-clinical trials. Advanced research is still required to assess the therapeutic effectiveness of the known organic products and their isolated compounds in HD experimental models. Summary: Degeneration of neurons in Huntington’s disease is distinguished by progressive loss of motor coordination and muscle function. This is due to the expansion of CAG trinucleotide in the first exon of the htt gene responsible for neuronal death and neuronal network degeneration in the brain. It is believed that the factors such as molecular genetics, oxidative stress, excitotoxicity, mitochondrial dysfunction, neuroglia dysfunction, protein aggregation, and altered UPS leads to HD. The defensive effect of the natural product provides therapeutic efficacy against HD. Recent reports on natural drugs have enlightened the protective role against HD via antioxidant, anti-inflammatory, antiapoptotic, and neurofunctional regulation.