Inhibition of osteosarcoma cell proliferation by the Hedgehog-inhibitor cyclopamine.

Osteosarcomas (OS) are the most frequent primary malignant bone tumors in humans. Even though OS are chemosensitive, about 30% of patients must be considered poor responders and consequently have a dismal long term prognosis. The Hedgehog (Hh) gene is crucial in the signalling pathways of proliferation and differentiation during embryonic development. There is evidence that uncontrolled activation of this pathway results in specific types of cancer and that inhibition of Hh signalling is able to suppress tumour growth and to induce apoptosis of neoplastic cells. This study investigates the impact of the steroidal alkaloid and Hh-inhibitor cyclopamine on osteosarcoma cells. Thus we demonstrate the drug's impact on cellular proliferation, cell cycle cell death as well as the cells' metabolism. We here demonstrate that cyclopamine exhibits a high efficacy against the osteosarcoma cell lines HOS, SaOS and OS-KA, a self-established primary osteosarcoma cell line. In particular, cyclopamine is able to inhibit proliferation and to promote cell death. Our results provide evidence for the potency of the Hh-inhibitor cyclopamine as a future treatment of osteosarcomas.

[Hypomelanosis Ito in translocation trisomy 9/mosaicism (46,XX/46,XX,t(9;9)(p24;p24)). Spontaneous remission in childhood]. / Hypomelanosis Ito bei Translokationstrisomie 9/Mosaik (46,XX/46,XX, t(9;9)(p24;p24)). Spontane Rückbildung im Kindesalter.

A 4 5/12 years old girl with hypomelanosis of Ito (HI) presented on the 3rd day of life with hypopigmented streaks and whorls following the lines of Blaschko on the back, the arms and the legs. In addition, patchy depigmented areas were present on the trunk. Extracutaneous manifestations included dystopia of the right kidney, atrial septal defect, persistent ductus arteriosus, hearing impairment, EEG abnormalities, and asymmetric dilatation of the ventricle system and a vermal atrophy as documented in the MRT of the brain. Cytogenetic analysis showed a mosaic 46,XX/46,XX,t(9;9)(p24;p24) present in the lymphocytes and skin fibroblasts. The mother's karyotype in her lymphocytes was normal. At reexamination at the age of 4 5/12 years the girl was retarded. In particular, the speech development was severely delayed. Interestingly we found only very small areas of hypopigmentation, which, unless one knew the previous findings, would not have been diagnostic.

Prenatal diagnosis of a fetus with a cryptic translocation 4p;18p and Wolf-Hirschhorn syndrome (WHS).

Wolf-Hirschhorn Syndrome (WHS) is caused by distal deletion of the short arm of chromosome 4 and is characterized by growth deficiency, mental retardation, a distinctive, 'greek-helmet' facial appearance, microcephaly, ear lobe anomalies, and sacral dimples. We report a family with a balanced chromosomal translocation 4;18(p15.32;p11.21) in the father and an unbalanced translocation resulting in partial monosomy 4 and partial trisomy 18 in one living boy and a prenatally diagnosed male fetus. Both showed abnormalities consistent with WHS and had in addition aplasia of one umbilical artery. Karyotyping of another stillborn fetus revealed a supernumerary derivative chromosome der(18)t(4;18)(p15.32;p11.21) of paternal origin and two normal chromosomes 4. The umbilical cord had three normal vessels. A third stillborn fetus with the same balanced translocation as the father had a single umbilical artery and hygroma colli.

Detection of TSPY protein in a unilateral microscopic gonadoblastoma of a Turner mosaic patient with a Y-derived marker chromosome.

Gonadoblastomas are seen almost exclusively in dysgenetic gonads of patients with a chromosomal mosaicism of 45,X and an additional Y-bearing cell line. This paper presents a case of a Turner mosaic patient with 45,X/46,X,+mar karyotype, who developed a unilateral microscopic gonadoblastoma. Cytogenetic and molecular analysis confirmed a Y-chromosomal origin of the marker chromosome, with a deletion of the distal Yq arm and the proposed region of a so far undefined gonadoblastoma locus (GBY) present. One of the candidate genes within the postulated GBY region is TSPY (testis-specific protein Y-encoded). To study the TSPY protein expression, an anti-fusion protein antibody was used for immunohistochemistry of the patient's gonads. In contrast to the dysgenetic gonad, an intense immunoreaction was found in gonadoblastoma tumour cells of the other gonad. These results confirm the high level of TSPY protein expression by these cells and demonstrate the value of this antibody as an additional marker to confirm the diagnosis of gonadoblastoma.

Translocation trisomy dup(21q) and free trisomy 21 can be distinguished by interphase-FISH.

The possibility of distinguishing in routine diagnostics translocation trisomy dup(21q) from disomy 21 as well as from free trisomy 21 using interphase fluorescence in situ hybridization (FISH) with a single copy probe (LSI 21) localized on chromosome 21q22.13-q22.2 is described. In free trisomy 21 and translocation trisomy dup(21q) 94%-98% of the nuclei exhibit 3 specific signals, while in disomy 21 only up to 6% of them have 3 false positive signals. Furthermore, reliable differentiation between free and translocation trisomy dup(21q) can be achieved by evaluating the percentage of nuclei with one single and two co-localized chromosome 21q22.13-q22.2 specific signals in 50-100 interphase nuclei. While in translocation trisomy 75+/-4.3% are co-localized due to a chromosomal rearrangement, in free trisomy 21 only 40+/-2.83% of the nuclei have two co-localized signals by chance. No differences in interphase signal distribution could be detected in two cases with a dicentric chromosome dup(21q) compared to one case with a monocentric one, a comparison not previously carried out. In addition, the single copy probe LSI 21 was compared with the alphoid probe D13Z1/D21Z1 which was found to be unsuitable for such assays due to polymorphisms in the á satellite regions of chromosome 21.

Distribution of marker-Y chromosome containing cells in different tissues of a Turner mosaic patient with mixed gonadal dysgenesis.

We here describe a 12-year-old girl with numerous Turner stigmata and virilized external genitalia. Chromosome analysis of PHA stimulated lymphocytes using different banding techniques revealed a 45,X/46,X,+mar Turner mosaicism with the prominent marker present in about 90% of the blood cells. A PCR-based analysis using a set of 9 STS from different regions of the human Y chromosome indicated the presence of Y chromosomal material with a deletion breakpoint most likely within deletion interval 6. Because of the risk of gonadoblastoma for Turner patients carrying Y chromosomal material, and clinical indications of functional testicular tissue, a gonadectomy in addition to surgical correction of the external genitalia was performed. The histological analysis of the gonads showed a mixture of testicular tissue and ovarian stroma, thus indicating mixed gonadal dysgenesis. Fibroblasts from skin and different parts of the gonads were cytogenetically analyzed and showed a variable distribution of the Y-derived marker between 4% in skin, 11-31% in gonadal tissue and up to 90% in peripheral lymphocytes.

Pierre Robin sequence and hyperphalangy--a genetic entity (Catel-Manzke syndrome).

Since the first description by Catel and Manzke of hyperphalangy and clinodactyly of the index finger associated with Robin malformation sequence, seven further cases have been published. In two families more than one case occurred. Another family is presented with possibly two affected boys showing variable features of the syndrome. It is proposed that the trait is X-chromosomal, recessively inherited.

HLA-B27-positive oligarthritis.

68 cases with polyarthritis were selected from 406 HLA B 27 positive patients with various rheumatic diseases excluding ankylosing spondylitis (AS) or Reiter's disease. 23 fulfilled at least 5 criteria of the ARA for the diagnosis of rheumatoid arthritis (RA). 5 suffered from polyarthritis and psoriasis. The remaining 40 patients expressed an asymmetric oligarthritis especially of the lower limbs (knee, ankle) affecting predominantly young adult men. Sacroiliitis was observed in 10 cases. Joint erosions, rheumatoid factors and visceral manifestations were uncommon. The arthritic pattern of B 27 positive oligarthritis differed clearly from rheumatoid arthritis (n = 34) and psoriatic arthritis (n = 15), but was similar to peripheral joint involvement in AS (n = 32) except for the higher incidence of coxitis in AS. HLA typing is helpful not only in the early diagnosis of AS but also in the differential diagnosis of unclassifiable polyarthritis.

[Decrease of mitogen stimulation of lymphocytes in rheumatoid arthritis]. / Verminderung der Mitogenstimulation von Blutlymphozyten bei der rheumatoiden Arthritis.

Peripheral blood lymphocytes of 51 patients with rheumatoid arthritis and of 14 healthy blood donors were cultivated in autologous and homologous normal pool-serum and stimulated with phytohemagglutinin (PHA-P), concanavalin A (Con A) and pokeweed mitogen (PMW) in different doses. In autologous as well as in homologous pool-serum the 3H-thymidine uptake of stimulated lymphocytes from patients with rheumatoid arthritis was significantly depressed independent of the mitogen concentration used. Serum from patients with rheumatoid arthritis depressed the activation of blood donor lymphocytes by Con A and PWM. The depression of lymphocyte activation was not related to the presence of rheumatoid factors in the serum, the severity of the disease and the anti-rheumatic therapy.