Serpentine fibula — polycystic kidney syndrome and Melnick-Needles syndrome are different disorders

We report on the third patient with serpentine fibula-polycystic kidney syndrome. Main features in the three reported cases were growth retardation, abnormal face, hirsutism, short neck, bowed forearms and lower legs due to bowed radii and elongated serpentine fibulae, and metatarsus adductus. Two patients including our own were deaf. All were mentally normal, all were female and sporadic. In addition, we report on a girl with Melnick-Needles syndrome and illustrate the similarities and differences between these syndromes.     

Detection of human cytomegalovirus in urine samples by cell culture,early antigen assay and polymerase chain reaction

Summary With the advent of effective therapy rapid, sensitive and reliable assays for diagnosis of human cytomegalovirus (HCMV) infections are required. In a total of 1,928 urine samples, detection of HCMV-immediate early antigen in a spin amplified microplate culture by a monoclonal antibody and immunoperoxidase staining (EA-assay) was compared with virus isolation in cell culture. Sensitivity of the EA assay was 85.5% and specificity was 99.5% compared with virus isolation. Overall agreement of both assays was 97.8%. In addition, in 235/1,928 urine samples amplification of HCMV-DNA was performed by means of polymerase chain reaction (PCR) using primers from the immediate early (IE1) gene region and 141/1,928 using primers from the late region (LA). The sensitivity of PCR compared with virus isolation was 67.8% for IE1 primers and 94.1% for LA primers (statistical significance: p<0.01, Chi-square-test). Overall agreement between virus isolation and PCR was 88.5% for IE1-PCR and 84.4% for LA-PCR. Discordant results were more often found in adults with acute infection and immunocompromised patients than in infants.

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Nachweis von humanem Zytomegalievirus in Urinproben mittels Zellkultur, Early-Antigen-Test und Polymerasekettenreaktion

Zusammenfassung Schnelle und verläßliche Teste für die Diagnostik von Zytomegalie- (CMV) Infektionen sind besonders seit dem Aufkommen einer effektiven antiviralen Therapie gefordert. Bei insgesamt 1928 Urinproben wurde der Nachweis von CMV-Frühantigen in Zentrifugationskultur (Mikrotiterplatte) mit einem monoklonalen Antikörper und Immunperoxidase-Färbung (Early-Antigen-Test) mit der herkömmlichen Virusisolierung in Zellkultur verglichen. Die Sensitivität des Early-Antigen-Tests betrug 85,5%, die Spezifität 99,5% im Vergleich zur Virusisolierung. Die Übereinstimmung beider Tests war 97,8%. Zusätzlich erfolgte in 235 der 1928 Urinproben der Nachweis von CMV-DNS mittels Polymerase-Kettenreaktion (PCR) mit Primern aus dem Bereich des viralen Immediate-Early-Gens (IE1) und in 141 der 1928 Proben mit Primern aus dem Bereich des Late-Gens (LA). Im Vergleich zur Virusisolierung erreichte die PCR eine Sensitivität von 67,8% mit IE1-Primern und von 94,1% mit LA-Primern (statistische Signifikanz: p<0.01, Chi-square-Test). Die Übereinstimmung von Virusisolierung und PCR betrug 88,5% (IE1) und 84,4% (LA). Bei Erwachsenen mit akuter Infektion und Immunsupprimierten Patienten waren die Ergebnisse weniger übereinstimmend als bei Kindern.

     

Mediators of antigen-induced gastrin release: role of antigen-antibody complexes and the complement system

That orally administered antigen was shown to induce gastrin release in immunized animals was a new aspect of gastrointestinal physiology. The mediators responsible for this immunological effect are still unclear. In an attempt to discover more about the mechanisms regarding antigen-induced gastrin release, we developed an in vitro system where fragments of rat antral mucosa were challenged. This makes it possible to determine the role of antigen-antibody complexes and the complement system in the mechanism of antigen-induced gastrin release. Wistar rats were immunized in vivo with NIP-OVA and mucosal fragments were challenged in vitro with NIP-HGG. Gastrin was determined after a preincubation and a challenged incubation period without supernatants. After antigenic challenge, supernatants were used for in vitro challenge in order to rule out the presence of a soluble mediator and activation of complement. In a second group of experiments Wistar rats were used to study in vitro the release of specific antibodies after antigenic challenge. With this experimental design we were able to show increased gastrin secretion after antigenic challenge in vitro in the presence of intact tissue. It is shown that the increased gastrin release is most probably mediated by activation of the complement system in the presence of antigen-antibody complexes. These are built up by specific anti-NIP antibodies and NIP-HGG used for the challenge. The complement system might be the final pathway of the observed in-creased gastrin release.     

Connatal varicella embryo-fetopathy

Connatal varicella embryo-fetopathy is rare. A newborn is described with severe cutaneous lesions, contractures, and hypoplasia of limbs following maternal varicella at the 13th week of pregnancy. Persisting low CF- and ELISA IgG-antibodies up to the age of 18 months give evidence for a prenatal varicella infection. Varicella contact during pregnancy requires rapid serodiagnosis of the immune status; if the women are seronegative, the application of zoster-hyperimmunoglobulin is recommended.     

Junctional complexes in the preimplantation rabbit embryo.

The morphology and development of junctional complexes between blastomeres of the preimplantation rabbit embryo were investigated using several approaches. Electron microscopic examination of embryos stained en bloc with uranyl acetate, and the study of junction permeability using horseradish peroxidase and lanthanum nitrate provided information on structure, intermembrane spacing and permeability of the junctional complexes. In addition, the freeze fracture technique was used with day 5 and day 6 blastocysts, since the large size of these embryos facilitated use of this method. These experiments showed that although rudimentary junctions were present between blastomeres of the early cleavage stages, effective tight junctions were not present until the blastocyst stage. Electron microscopic examination of thin sections revealed apical foci of membrane approximation or "fusion" between trophoblast cells by day 4. Freeze fracturing revealed a lattice of interconnecting ridges (on the A face) and grooves (on the B face) in the apical region between trophoblast cells of the day 5 blastocyst. This lattice formed a continuous band along the apical margin of each cell, and therefore constituted a zonula occludens. The zonula occludens of the day 5 blastocyst averages 2-3 ridges per lattice, while day 6 blastocysts had lattices that averaged 5-6 ridges. Also seen in the freeze fracture replicas from the day 5 and day 6 blastocysts were local accumulations of intramembranous particles on the A face. These particles were often observed in aggregates similar to those of previously described gap junctions. It could not be determined whether these small regions of particles were true gap junctions or a possible primitive form of gap junction because the complementary pitted surfaces (B face pits) were not demonstrated.     

Gastric cancer cell detection in peritoneal washing: cytology versus RT-PCR for CEA transcripts.

This study investigates the sensitivity and specificity of cytology, qualitative, and real-time RT-PCR methods in free cancer cell detection of peritoneal washing from gastric cancer patients. Peritoneal washings were collected from 65 gastric cancer patients for routine cytology and total RNA extraction for qualitative and real-time RT-PCR for CEA. The sensitivity and false-positive rate was 51.1%, 0% for cytology, 48.9% and 5% for qualitative RT-PCR for CEA, and 42.5% and 5% for real-time RT-PCR for CEA. The qualitative and real time RT-PCR results show high concordance rate (89.7%). The highest sensitivity was obtained by the combination of cytology with qualitative RT-PCR for CEA (70.2%). RT-PCR results were positive in 63.6% of cytologic "atypia" cases. Combination of cytology and either of the RT-PCR methods resulted in significantly higher sensitivity than any one of the three methods alone (P < 0.05). There was no definite advantage of the real-time RT-PCR over the conventional RT-PCR.     

Rapid typing of the codon 129 polymorphism of the human prion protein gene by combined real-time PCR and melting curve analysis

Homozygosity of methionine (m/m) at amino acid residue 129 (codon 129) of the human prion protein (PrP) has been reported for all so far analyzed cases of the new variant Creutzfeldt-Jakob disease (vCJD). This contrasts with its general distribution in the healthy Caucasian population of only about 43%. For this reason a predisposition for carriers of the corresponding genotype to develop vCJD after infection with the bovine spongiform encephalopathy (BSE) agent is assumed, and PCR based methods such as allele-specific oligonucleotide hybridization or restriction analysis and sequencing have been developed for codon 129 genotyping. These methods are cumbersome and time-consuming and the need for extensive post-amplification manipulations increases the risk of carry-over contamination with amplified products. To overcome these shortcomings, the authors developed a real-time PCR assay on the LightCycler (LC) instrument combining PCR and temperature melting curve analysis (Tm) in a closed vessel format. Forty-six swabs and blood samples from healthy donors were tested. Of these 23 (50%) were heterozygous at codon 129, 4 (8.7%) homozygous for valine and 19 (41.3%) homozygous for methionine. Accuracy of LC-genotyping was confirmed by automated sequencing of the amplified products. Taken together, genotyping of the codon 129 polymorphism by combined LC-PCR and melting-curve analysis with the LC-instrument is a reliable and easy to perform method even in a screening context with numerous samples. Results can be obtained within 2 hours, including sample preparation.