Juri Kopolovic

Standardization of the Teratoma Assay for Analysis of Pluripotency of Human ES Cells and Biosafety of Their Differentiated Progeny

Teratoma tumor formation is an essential criterion in determining the pluripotency of human pluripotent stem cells. However, currently there is no consistent protocol for assessment of teratoma forming ability. Here we present detailed characterization of a teratoma assay that is based on subcutaneous co-transplantation of defined numbers of undifferentiated human embryonic stem cells (hESCs) with mitotically inactivated feeder cells and Matrigel into immunodeficient mice. The assay was highly reproducible and 100% efficient when 100,000 hESCs were transplanted. It was sensitive, promoting teratoma formation after transplantation of 100 hESCs, though larger numbers of animals and longer follow-up were required. The assay could detect residual teratoma forming cells within differentiated hESC populations however its sensitivity was decreased in the presence of differentiated cells. Our data lay the foundation, for standardization of a teratoma assay for pluripotency analysis. The assay can also be used for bio-safety analysis of pluripotent stem cell-derived differentiated progeny.

Alveolar soft-part sarcoma of the female genital tract

SummaryAn alveolar soft-part sarcoma of the uterine cervix observed in a 26-year-old women is described. One year after extended radical hysterectomy there was no evidence of tumor recurrence. To the best of our knowledge, this is the second reported case of alveolar soft-part sarcoma arising in the female genital tract and the first description of this tumor in the uterine cervix.

Familial Mediterranean Fever (FMF) with Proteinuria: Clinical Features, Histology, Predictors, and Prognosis in a Cohort of 25 Patients

Objective. Reactive (AA) amyloidosis may complicate familial Mediterranean fever (FMF), the prototype of autoinflammatory diseases. Thus, proteinuria in FMF is commonly viewed as resulting from amyloidosis, and kidney biopsy is deemed superfluous. However, nephropathy other than amyloidosis has been described in FMF, but its rate and distinctive characteristics are unknown. Our aim was to determine the rate and underlying pathology of FMF-related nonamyloidotic proteinuria and compare its clinical course, demographic, and genetic features to those of FMF-amyloid nephropathy. Methods. This study is a retrospective analysis of data from patients with FMF undergoing kidney biopsy for proteinuria above 0.5 g/24 h, over 10 years (2001–2011). Clinical, laboratory, genetic, and pathology data were abstracted from patient files. Biopsies were viewed by an experienced pathologist, as necessary. Results. Of the 25 patients referred for kidney biopsy, only 15 (60%) were diagnosed with amyloid kidney disease (AKD), and 10 were diagnosed with another nephropathy. The AKD and nonamyloid kidney disease (NAKD) groups were comparable on most variables, but showed distinct characteristics with regard to the degree of proteinuria (6.45 ± 4.3 g vs 2.14 ± 1.6 g, p = 0.006), rate of severe FMF (14 vs 5 patients, p = 0.022), and rate of development of end stage renal disease (73.3% vs 20%, p = 0.015), respectively. Conclusion. NAKD is common in FMF and, compared to amyloidosis, it is featured with milder course and better prognosis. Contrary to common practice, it is highly recommended to obtain a kidney biopsy from patients with FMF and proteinuria more than 0.5 g/24 h.

Platelet abnormalities in Down's syndrome

Nine individuals with Downs syndrome and 12 normal children were studied. Platelets from Downs syndrome (DS) were smaller than normal and had reduced numbers and volumes of electron dense bodies. Whole cell and dense body calcium levels were lower than normal, while platelet surface net negative charge and anionic sites were found to be elevated in DS platelets. The findings suggest a calcium‐related membrane abnormality as one of the defects in DS platelets.

Development of Hodgkin’s Disease in a Patient with Leprosy

We present a patient with leprosy who developed Hodgkins disease of the nodular sclerosing type. There are two previous reports describing the combination of leprosy and Hodgkins disease in a single patient [3, 9]. Hodgkins disease was diagnosed 14 months after the complete disappearance of mycobacterium leprae from the skin lesions, under treatment with DDS (diamino-diphenyl-sulfone). Hodgkins disease was treated by irradiation and chemotherapy. Obstructive jaundice developed which resolved under treatment by irradiation of the hilar area of the liver, chemotherapy and hormones. During two years of immuno-suppressive therapy, without DDS, no exacerbation of the leprosy occurred.

Mutations in the familial Mediterranean fever gene of patients with IgA nephropathy and other forms of glomerulonephritis

Glomerulonephritis, particularly IgA nephropathy (IgAN), seems to be more common in familial Mediterranean fever (FMF), an inherited disease caused by mutations in the MEditerranean FeVer gene (MEFV). The present study is aimed to determine, in populations not suffering from FMF, whether carriage of MEFV mutations may modify or precipitate IgAN and other forms of primary glomerulonephritis (PGN). Forty patients with biopsy proven IgAN and 40 with PGN were surveyed for the presence of the three most common MEFV mutations (M694V, V726A and E148Q), using polymerase chain reaction amplification and restriction enzyme analysis. The rate of MEFV mutations in the patients was related to the expected carrier rate in the general population of the same ethnic extraction. The effect of mutation carriage on the disease course was determined in the IgAN patient group. The frequency of MEFV mutations in IgAN or PGN was comparable to that found in ethnically adjusted general population (pu2003=u20030.1 and 0.5, respectively). Carriage of mutated MEFV was not associated with the course and severity of the disease or findings in kidney biopsy and urine analysis. In a population, mostly of Jewish extraction, MEFV mutations do not seem to predispose to the development of IgAN and other forms of PGN or affect the phenotype.

P01-030 – Proteinuria in FMF – prediction of nephropathy type

Reactive (AA) amyloidosis may complicate Familial Mediterranean fever (FMF), the prototype of autoinflammatory diseases. Thus, proteinuria in FMF is commonly viewed as resulting from amyloidosis and kidney biopsy is deemed superfluous. However, nephropathy other than amyloidosis has been described in FMF, but its rate and distinctive characteristics are unknown.