Preliminary study of the incidence of disomy in sperm fractions after MicroSort flow cytometry

Using triple colour fluorescent in-situ hybridization (FISH) we have evaluated, on a blind basis, the disomy level for chromosome 21 and the sex chromosomes in flow cytometric sorted sperm samples. There were no statistically significant differences in the disomy rates when comparing the sorted samples (either for X- or Y-bearing spermatozoa) with non-sorted samples. There were no diploid spermatozoa observed in any sample group after MicroSort processing.     

Short-term and long-term results after early treatment with the castillo morales stimulating plate

The present study is divided into 2 parts: The first explains the concept of stimulating plate therapy at the Muenster Clinic and the second presents a longitudinal study of mouth and tongue posture in trisomy 21 children treated with this concept.47 children underwent a pre-treatment examination (Rec1), and 38 attended a first follow-up examination (Rec2) after an average of 4 months of stimulating plate treatment. The second follow-up examination (Rec3) was on average 53 months after the end of treatment and covered 18 patients. Data acquisition was based on standardized parent questionnaires and standardized clinical examination records. Rec2 showed in part a highly significant improvement in orofacial appearance. These results were confirmed by the parent interview, according to which the mouth and tongue posture improved in 76.3% of the children during the time they were wearing the plate. Even when the plate was not in place, the result remained stable in 65.8% of the patients. Up to Rec3, further improvement was recorded in some results. The symptoms: mouth mostly wide open and tongue mostly protruding way over lips were reduced. The number of children whose tongue was mostly in the oral cavity increased correspondingly.At Rec3, the parents also rated the mouth posture in 88.9% and the tongue posture in 77.7% of the children as stable or further improved.It can be concluded from the results of this study that stimulating plate treatment in combination with physiotherapy provides a long-term improvement in the mouth and tongue posture of children with Down's syndrome.     

P21 WAF1/Cip1 Gene Expression in Primary Human Hepatocellular Carcinoma and its Relationship with P53 Gene Mutation

Ｐ２１ＷＡＦ１／Ｃｉｐ１ｐｒｏｔｅｉｎ（ｐ２１）ｉｓｔｈｅｐｒｏｄｕｃｔｏｆｗａｆ１ｇｅｎｅ,ａｌｓｏｋｎｏｗｎａｓｃｉｐ１,ｓｄｉ１［１,２］．Ｐ２１ＷＡＦ１／Ｃｉｐ１ｉｓａｕｎｉｖｅｒｓａｌｉｎｈｉｂｉｔｏｒｏｆｃｙｃｌｉｎ－ｄｅｐｅｎｄｅｎｔｋ...     

Auswirkungen von Argon-Gasembolien während eines PneumoperitoneumsRID="*"ID="*"Die Studie wurde zum Teil gefördert von der DFG (Bo 1329/8–1)

Zusammenfassung. In einer experimentellen Studie wurde bei 10 Schweinen mit einem mittleren K?rpergewicht von 18,9 (15–24) kg eine intraven?se CO₂- oder Argon-Embolie mit 10, 20 und 30 ml Gas durchgeführt. Das invasive Monitoring zeigte bei der Gasembolie mit Argon im Gegensatz zur Gasembolie mit CO₂ einen st?rkeren Anstieg des pulmonal arteriellen Drucks (p < 0,001), einen st?rkeren Abfall des endexspiratorischen CO₂ (p < 0,01), des Herzminutenvolumens (p < 0,01) und des mittleren arteriellen Drucks (p < 0,01). In der Argon-Gruppe (n = 5) starben zwei Tiere nach 20 bzw. 30 ml Bolusgabe. Ein weiteres Tier konnte nach Gabe von 30 ml Bolus erfolgreich reanimiert werden. In der CO₂-Gruppe (n = 5) starb weder eines der Tiere noch war eine Reanimation erforderlich. Wenig l?sliche Gase wie Argon sollten in Situationen mit erh?htem Risiko einer Gasembolie nicht angewendet werden. ID=" Dr. T. Junghans Klinik f&uuml;r Allgemein-, Visceral-, Gef&auml;&szlig;- und Thoraxchirurgie Universit&auml;tsklinikum Medizinische Fakult&auml;t der Humboldt-Universit&auml;t Campus Charit&eacute; Mitte Schumannstra&szlig;e 20/21 D-10117 Berlin     

Value of direct measurement of active renin concentrations in congenital adrenal hyperplasia due to 21-hydroxylase deficiency

In congenital adrenal hyperplasia (CAR) due to 21-hydroxylase deficiency, measurement of plasma renin activity (PRA) has been the method of choice in diagnosing salt loss and in monitoring adequacy of mineralocorticoid replacement therapy. Due to methodological problems in PRA determinations, direct immunoradiometric assays for the measurement of active renin concentration have been developed. We measured PRA and active renin concentrations simultaneously in 39 patients with CAH (30 salt-wasting, 9 simple virilizing) to evaluate the potential role of this new method in the management of this disease. PRA was determined with an enzymatic assay (sample volume: 2 × 1000 l plasma), active renin concentration with a direct immunoradiometric assay (sample volume: 2 × 200 l plasma or serum). We found a highly significant correlation between active renin and PRA in our patients (P < 0.001), as previously shown in healthy subjects. Active renin was as reliable as PRA to assess the quality of mineralocorticoid replacement.     

Tetrahydronorharmane (Tetrahydro-β-carboline), a physiologically occurring compound of indole metabolism

Summary In the present paper a sensitive method is described to measure tetrahydronorharmane (THN) in the urine of man and rats as well as in the forebrain of rats. The compound is extracted into diethyl ether, separated by thin layer chromatography (TLC), acetylated with radiolabelled acetic anhydride and further isolated by two-dimensional TLC development. The existence of THN in urine of man was proven by using chromatography with different solvent systems, cocristallisation, isotope dilution technique as well as mass-spectrometry. The amount of THN in the urine varied over a wide range.With the same method it was demonstrated that THN occurs also in the forebrain of rats. The concentration increases after loading with tryptamine.The findings are discussed in view of the hypothesis that THN acts as a compound modulating neuronal mechanism.     

Family studies of the steroid 21-hydroxylase and coplement C4 genes define 11 haplotypes in classical congenital adrenal hyperplasia in The Netherlands

Two steroid 21-hydroxylase genes are normally present within the human major histocompatibility complex near the genes encoding the fourth component of complement (C4A and C4B). Steroid 21-hydroxylase is encoded by the CYP21 gene, while the highly homologous CYP21P gene is a pseudogene. We studied steroid 21-hydroxylase and complement C4 haplotypes in 33 Dutch patients (29 families) suffering form classical congenital adrenal hyperplasia (CAH) and in their 80 family members, and also in 55 unrelated healthy controls, using 21-hydroxylase and complement C4 cDNA probes. Eleven different haplotypes, defined in terms of gene deletions, gene duplications, conversions of CYP21 to CYP21P, and long and short C4 genes, were found. In 23% of the patients' haplotypes, the CYP21 gene was deleted; in 12%, it was converted into a CYP21P pseudogene. In the remaining 65%, the defect was apparently caused by a mutation not detectable by this method. The most common haplotype (with one CYP21 and one CYP21P gene) was significantly more often observed in patients with simple virilizing CAH than in those with salt-losing CAH. Comparison of the 21-hydroxylase haplotypes found in CAH patients from several countries shows evidence for considerable genetic variation between the groups studied.     

Molecular detection of genetic defects in congenital adrenal hyperplasia due to 21-hydroxylase deficiency: A study of 27 families

Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase (21-OHase) deficiency is inherited as an autosomal recessive trait. Patients can present with the salt wasting, simple virilizing or a non-classical form of the disease. The gene for P450C21, the enzyme carrying 21-OHase activity, has been mapped to the major histocompatibility complex on chromosome 6p. Using molecular hybridisation techniques we have studied the genetic defect in 27 families with one or more affected off-spring diagnosed and treated at the University Hospital of Essen. DNA samples were digested with restriction endonucleaseTaqI,PvuII,BglII, andEcoRI and analysed by Southern blot hybridisation with the cDNA probe pC21/3c. Eleven of 40 haplotypes associated with the salt wasting form were found to have a large deletion of 30 kb affecting the 5 end of the active 21-OHase gene and the 3 end of the closely linked pseudogene. Results in another 11 cases are compatible with gene conversion; 18 cases were not informative. The 30 kb deletion was associated with a combination of the HLA antigens Bw47 and DR7 in 7 of 11 cases. In the haplotypes with gene conversion, no linkage disequilibrium to HLA antigens was found. No apparent gene alterations were detected in simple virilizing and non-classical haplotypes. The direct detection of the genetic defect in 55% of the salt wasting haplotypes may help to improve predictive testing in families with CAH.     

葡萄胎p53、p21~(CIP1)及p185蛋白表达与恶变的关系

目的 :探讨葡萄胎p5 3、p2 1CIP1及p185蛋白表达与恶变的关系及其临床特点。方法 :免疫组化法检测葡萄胎标本中p5 3、p2 1CIP1及p185蛋白的表达 ,以侵蚀性葡萄胎及绒癌为对照 ,并回顾性分析其临床资料。结果 :葡萄胎组p5 3、p2 1CIP1、p185蛋白阳性表达率分别为 35 % (35 / 10 0 ) ,71% (71/ 10 0 )及 36 % (36 / 10 0 ) ,与恶性对照组相比均有显著性差异 (P <0 .0 5 ) ;而完全性与部分性葡萄胎 ,完全性葡萄胎恶变组与非恶变组之间差异均无显著性 (P >0 .0 5 ) ,但恶变组p2 1CIP1表达有降低趋势。结论 :p5 3、p2 1CIP1及p185蛋白表达改变与葡萄胎恶变无确定关系 ,但可作为晚期现象出现于恶性滋养细胞肿瘤中 ,其中p2 1CIP1蛋白表达降低提示滋养细胞有向恶性转化的倾向     

Utility of peptide–protein affinity complexes in proteomics: identification of interaction partners of a tumor suppressor peptide*

Abstract: We used a N‐biotinylated peptide analog of the C‐terminal domain of the tumor suppressor protein, p21^cip1/waf1 to elucidate peptide/protein interacting partners. The C‐terminal domain of p21^cip1/waf1 protein spanning 141–160 amino acid residues is known to bind PCNA and this interaction is important in many biological processes including cell‐cycle control. This C‐terminal 20‐mer efficiently extracts PCNA in the presence of a variety of N‐ or C‐terminally attached affinity tags. Using difference silver stained 2D gels combined with in‐gel tryptic digests, we identified the difference spots using MALDI‐TOF mass spectrometry‐based peptide mass fingerprinting followed by a database search using profound against NCBIs human nonredundant protein sequence data bank. Identified spots include the p48 subunit of chromatin assembly factor‐1, the heat shock 70 protein analog BiP, calmodulin, nucleolin and a spot similar in size to dimeric PCNA. In contrast, microcapillary ion‐trap LC‐MS/MS analysis of a tryptic digest of entire affinity extracts derived from both control and experimental runs followed by database searches using sequest confirmed the presence of most of the above proteins. This strategy also identified hnRNPA1, HPSP90α, HSP40 and T‐complex protein 1, a protein similar to prothymosin, and a possible allelic variant of the p21^cip1/waf1 protein. The use of N‐biotinylated peptide derived from the C‐terminal domain of p21^cip1/waf1 protein in proteomic analysis exemplified here suggests that peptides obtained from intracellular functional screens could also potentially serve as efficient baits to discover new drug targets.