Defining the limits of detection for chromosome rearrangements in the preimplantation embryo using next generation sequencing.

STUDY QUESTION: Is next generation sequencing (NGS) capable of detecting smaller sub-chromosomal rearrangements in human embryos than the manufacturer's quoted resolution suggests? SUMMARY ANSWER: NGS was able to detect unbalanced chromosome segments smaller than the manufacturer's resolution. WHAT IS KNOWN ALREADY: Array Comparative Genomic Hybridization (array-CGH) has been the gold standard platform used for PGD of chromosome rearrangements. NGS is a viable alternative to array-CGH for PGD of chromosome arrangements given that the manufacturer's guidelines quote a resolution of ≥20 Mb. However, as many patients carry a chromosome rearrangement <20 Mb, the detection limits of NGS warrant further investigation. STUDY DESIGN, SIZE, DURATION: This study involved a retrospective assessment of stored DNA samples from embryos that had previously been diagnosed as unbalanced by array-CGH as part of routine PGD in two separate IVF clinics between November 2013 and April 2017. SurePlex whole genome amplification (WGA) products derived from DNA extracted from an embryo biopsy sample known to carry an unbalanced form of a chromosome rearrangement were subjected to a specific NGS workflow (VeriSeq PGS). The results from the two technologies were compared for each sample. PARTICIPANTS/MATERIALS, SETTING, METHODS: WGA products from 200 embryos known to carry unbalanced rearrangements were sequenced and analysed. These embryos had been created by 75 patients known to carry a chromosome rearrangement (68 reciprocal translocations, 3 pericentric inversions, 1 paracentric inversion, 2 insertions and 1 dual reciprocal and inversion). Each sample was assessed for the size of the segmental gain/loss (Mb), copy number for each segment and chromosome, segregation pattern, the number of bins in the analysis software used and concordance with array-CGH results. MAIN RESULTS AND THE ROLE OF CHANCE: A total of 294 unbalanced chromosome segments were assessed. NGS was capable of detecting 285/294 (97%) unbalanced segments previously identified using array-CGH. The final PGD diagnosis was concordant for 200/200 (100%) embryos. In total, 44/75 (59%) patients contained an unbalanced chromosome segment below the quoted 20 Mb manufacturer's stated resolution. Of these, 35/44 (80%) patients had segments that were able to be detected using NGS, whilst maintaining clinical outcome concordance. LIMITATIONS, REASONS FOR CAUTION: Our study subset did not include any rearrangements involving the Y chromosome. NGS has less available bins per chromosome compared to the array-CGH platform used, thus it remains possible that chromosome rearrangements predicted to be small but still detectable by array-CGH may not be feasible for testing using NGS. This should be considered when undertaking a theoretical feasibility assessment for detecting the chromosome rearrangement in question. Only one specific workflow for WGA and NGS was investigated in this study. WIDER IMPLICATIONS OF THE FINDINGS: This study has shown that NGS is available for the detection of unbalanced chromosome rearrangements ≥10 Mb. STUDY FUNDING/COMPETING INTEREST(S): Part sponsorship of the VeriSeq PGS kits used was provided by Illumina. The remainder of the kits were provided by two commercial IVF clinics. None of the authors has any conflicting interests to declare. TRIAL REGISTRATION NUMBER: N/A.

Myc--what we have learned from flies.

The Myc family proteins are key regulators of animal growth and development. dMyc, the only Drosophila member of the Myc gene family, is orthologous to the mammalian c-Myc oncoprotein. Extensive studies have revealed much about both upstream regulators and downstream target genes in the sphere of Myc regulation. Here, we review some of the critical discoveries made using the Drosophila model, in particular those studies that have explored the essential role of the Myc family in growth and cell cycle progression and identified many of the upstream signals and downstream targets common to both c-Myc and dMyc.

Newborn intraabdominal cystic lymphatic malformations.

Cystic lymphatic malformations are rare causes of abdominal masses in the newborn. Also known as mesenteric, omental, or retroperitoneal cysts, they can present in a variety of ways including, intestinal obstruction, volvulus, nonspecific abdominal pain, intracystic hemorrhage, or as an asymptomatic abdominal mass. Abdominal ultrasound scan provides a definitive diagnosis in most suspected cases. Complete resection is possible in most patients except those with extensive retroperitoneal involvement. Recurrence is unusual when complete resection is accomplished. Because most case series with complete data suggest that these entities are lymphatic malformations, the authors suggest the more specific term, cystic lymphatic malformations, be used to describe these lesions.

Hepatobiliary dysfunction as the initial manifestation of disseminated cryptococcosis.

A case of hepatobiliary dysfunction as the initial manifestation of disseminated cryptococcosis is described. The patient was admitted with symptoms of hepatitis with cholestatic jaundice. Antibody tests for hepatitis B and C and human immunodeficiency virus were negative. The patient continued to deteriorate clinically. Eventually, the patient succumbed to hepatic failure. Autopsy disclosed systemic cryptococcosis that caused extensive necrosis of the liver. In review of the literature, only nine cases of cryptococcal infection presenting as hepatitis, cholangitis, and cholecystitis as initial manifestation were reported. Four of these patients had been subjected to exploratory laparotomy for clinical suspicion of acute abdomen. One patient developed cirrhosis as a result of cryptococcal hepatitis. Two patients succumbed to hepatic failure. Cryptococcosis is known to occur commonly in immunocompromised patients, yet only two reported cases presenting as hepatitis were associated with immunocompromised status.

Acquired cystic disease of kidney associated with renal cell carcinoma in chronic dialysis patients.

We describe 2 cases of acquired cystic disease of the kidney (ACDK) associated with renal cell carcinoma in patients treated with long-term hemodialysis. Both patients have had dialysis for five and eight years, respectively. Renal cell carcinomas of these patients are small, averaging 2 cm in diameter. They are clear cell type. Atypical epithelial hyperplasia arising from cystic areas can be seen intermingling with carcinoma. This confirms that atypical epithelial hyperplasia is a precursor of renal cell carcinoma. Although the incidence of renal cell carcinoma arising in ACDK is on the rise, the issue of how to manage patients with ACDK remained unsettled and required further study.

Gastric infarction.

We have described a patient with an acute condition of the abdomen who had infarction of the stomach and the small intestine due to atheromatous thrombus of celiac and superior mesenteric arteries. We believe this unusual simultaneous occurrence of gastric and small intestinal infarction is coincidental. The outcome of gastric infarction is frequently fatal.

Small cell carcinoma of urinary bladder. Ultrastructural study.

Two cases of small cell carcinoma of the urinary bladder are presented. On gross examination they were found to be polypoid and shared light microscopic features of oat cell carcinoma of the lung. Ultrastructurally, however, they did not reveal features of neuroendocrine cells containing neurosecretory granules. In regard to histogenesis, they appear to arise from undifferentiated cells present in the transitional epithelium.

Stone-like calcification of hypernephroma.

A case of stone-like calcification of renal cell carcinoma is presented. The pattern of calcification in relation to prognosis of renal cell carcinoma is discussed.

Malignant fibrous histiocytoma of the ileum.

Malignant fibrous histiocytoma occurs most commonly in the extremities and trunk, but rarely in visceral organs. This report documents a case of malignant fibrous histiocytoma arising in the terminal ileum. Following surgical resection, there is no evidence of recurrence or metastasis in this patient after one-year follow-up studies.

CK-MB isoenzyme in patients with polymyositis.

Total serum creatine kinase and isoenzyme levels were studied in five patients with polymyositis. In all five patients, CK-MB isoenzyme was demonstrated by the column chromatography and electrophoretic method, although there was no evidence of myocardial infarction. The persistent elevation of CK-MB in patients with polymyositis is in contrast to the usual transient increase in myocardial infarction. Serial CK-MB isoenzyme quantitation can be used to distinguish myocardial infarction from polymyositis. CK-MB is a more sensitive indicator than AST and LDH as a monitoring device, but offers no advantage over total CK activity.

Case report. Pulmonary dirofilariasis in man.

A case of pulmonary dirofilariasis in a 69-year-old woman is presented. A review of the literature revealed 56 documented cases. As in most cases of pulmonary dirofilariasis, our patient presented roentgenologically a single noncalcified coin lesion and required thoracotomy for diagnosis. The gross and histologic characteristics of the lesion, the life cycle of the parasite, the subcutaneous form of human dirofilariasis, and the possible role of the immunologic tests in establishing the diagnosis are briefly discussed. The incidence of pulmonary dirofilariasis appears to be increasing. Awareness that such entity exists is important since correct preoperative diagnosis can alleviate unnecessary surgical intervention.

Angioimmunoblastic lymphadenopathy in a patient taking diphenylhydantoin.

The premise that chronic antigenic stimulation may be involved in lymphoproliferative disorders was considered in a patient with angioimmunoblastic lymphadenopathy who had received diphenylhydantoin (DPH) for seizure. This patient had clinical features overlapping with systemic lupus erythematosus (SLE) and serum antibody directed against DPH. It was proposed that the syndrome was caused by chronic antigenic stimulation with DPH.

Superficial spreading adenocarcinoma of appendix, cecum, and terminal ileum.

A case of superficial spreading adenocarcinoma involving terminal ileum, appendix and cecum is presented. The tumor might primarily arise from the proximal part of the appendix with extension to the cecum and terminal ileum, although multicentric origin of the tumor cannot be excluded. The mechanism underlying superficial infiltration of the tumor is through close contact and intramucosal lymphatic spread.

Budd-Chiari syndrome in women taking oral contraceptives.

The report concerns a 21-year-old woman who, after having taken oral contraceptives for three months, developed the Budd-Chiari syndrome as the results of a widespread chronic obliterative process involving the intrahepatic venous system. The diagnosis was established by liver biopsy. She was treated with supportive measures and completely recovered after 28 days of hospitalization. The literature is reviewed regarding the Budd-Chiari syndrome in women taking oral contraceptives and the clinical and pathological features seen in those reported patients are discussed.

PIP: Sine 1966 when the 1st reference of OCs (oral contraceptives) as a cause of Budd-Chiari syndrome was made, 18 such cases have been reported. A further case of a 21-year-old woman who, having taken OCs for 3 months, developed the Budd-Chiari syndrome as the result of a widespread chronic obliterative process involving the intrahepatic venous system is reported. The Budd-Chiari syndrome, i.e., occlusion of the hepatic veins, was diagnosed by liver biopsy. The treatment is described and recovery was complete following 28 days of hospitalization. The literature regarding the incidence of Budd-Chiari syndrome in women taking OCs is reviewed. In recent years, aggressive surgical treatment has been recommended for this syndrome.

Diffuse cystitis glandularis. Associated with adenocarcinomatous change.

Although cystitis glandularis has been considered a premalignant lesion, the instance of cystitis glandularis progression to adenocarcinoma or cystitis glandularis associated with adenocarcinomatous change is rare. This article includes 4 cases of neurogenic bladder with urinary diversion for different periods of time. In the first case with ureterocutaneostomy for twenty-five years diffuse cystitis glandularis with multifocal adneocarcinomatous change developed. The second case with suprapubic cystostomy for twenty-two years had diffuse cystitis glandularis of gastrointestinal type without evidence of malignancy. The other 2 cases with suprapubic cystostomy for merely ten years showed only mild to moderate cystitis glandularis and chronic cystitis with squamous metaplasia, respectively. The extent of cystitis glandularis appeared to correlate with the duration of urinary stasis. Ureterocutaneostomy rendered constant infection of the urinary bladder of the first case because of inadequate drainage. Thus, we assume that the intensity of the infection with a toxic product and virulence of organism may be responsible for the development of adenocarcinoma in this patient. Cystitis glandularis, especially diffuse type, can undergo malignant degeneration under constant irritation, but it is a long-term process.

Cholesterol granuloma of right testis.

A case of cholesterol granuloma of the testis is presented. This lesion might be caused by trauma and inflammation. The difference between cholesterol granuloma and lipogranuloma of the external genitalia is emphasized.