Somatic von hippel-lindau mutation in clear cell papillary cystadenoma of the epididymis

Papillary cystadenoma of the epididymis is an uncommon benign lesion that may occur sporadically or as a manifestation of von Hippel—Lindau (VHL) disease. Neither immunohistochemical studies nor molecular genetic analyses of the VHL gene have been reported previously for this lesion. The authors describe two cases of clear cell papillary cystadenoma of the epididymis, both of which were initially confused with metastatic renal cell carcinoma. Both lesions showed positive immunohistochemical staining for low and intermediate molecular weight keratins (Cam 5.2 and AE1/AE3), EMA, vimentin, α₁-antitrypsin, and α₁-antichymotrypsin. Each was negative for CEA. Because clear cell papillary cystadenoma is similar to renal cell carcinoma histologically, and because both occur as components of the von Hippel—Lindau disease complex, the authors analyzed both cases for the presence of mutations in the VHL gene. A somatic VHL gene mutation was detected in one of the two tumors by polymerase chain reaction followed by single-strand conformation polymorphism analysis. Direct sequencing revealed a cytosine to thymine transition at nucleotide 694, resulting in the replacement of an arginine with a stop codon after the sixth amino acid of exon 3. As the VHL gene is believed to function as a tumor suppressor gene, VHL gene mutations may play a role in the initiation of tumorigenesis in sporadic cystadenomas of the epididymis.     

Die Konzentration von Adenovirus-Antigenen mit Polyphosphors?ure

Zusammenfassung Mit Hilfe von Polyphosphaten lassen sich die Antigene von Adenovirus Typ 19 zeitsparend und schonend konzentrieren. In einem pH-Bereich zwischen 2,8 und 3,6 wurden bei Verwendung von 2 mg Polyphosphat je ml Antigen infektiöses Virus, Hämagglutinin und Adeno virus-Gruppen-antigen vollständig gefällt. Unter den gleichen Bedingungen wurde auch der größte Teil des Gesamtproteins und des komplementbindenden HeLa-Zellantigens präzipitiert, so daß man zwar eine Konzentrierung, aber keine Reinigung der virusspezifischen Komponenten erhält.

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Summary Antigens of adenovirus type 19 were concentrated by means of polyphosphate, a time-saving and mild procedure. Infectious virus, haemagglutinin and adenovirus group antigen were completely precipitated in a pH range between 2.8 and 3.6, when 2 mg. polyphosphate per ml. antigen was used. Under the same conditions, the bulk of total protein as well as of complement-fixing HeLa cell antigen was precipitated. Consequently, a concentration of virus-specific components was achieved, but no purification.

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Durchgeführt mit Unterstützung der Deutschen Forschungsgemeinschaft, Bad Godesberg.

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Herrn Prof. Dr.K. Herzberg zum 70. Geburtstag gewidmet.

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FrauI. Maurer und FräuleinI. Denker sei für ausgezeichnete technische Assistenz gedankt.     

In vitro adherence of radioactively labeled Escherichia coli in normal and cystitis-prone females.

Numerous investigators report data obtained using an in vitro quantitative assay for measuring bacterial adherence to epithelial cells. We found this assay to contain significant sources of error in the large variation in number of bacteria bound per cell and in the dependence on the investigator's visual counting of bacteria bound per cell. In the modified assay described here, we eliminated the need for visual counting of bacteria by incorporating the use of radioactively labeled Escherichia coli. This allowed quantitation of bacterial adherence to as many as 50,000 vaginal cells, whereas the visual counting system limits the determination to perhaps 50 cells. We feel that the use of radioactively labeled bacteria in place of the visual counting system increases the validity and sensitivity of this assay. Using the modified method, we found no statistically significant differences among values for adherence of E. coli type 04 to the vaginal cells of control and cystitis-prone women at either pH 6.4 or 4.0.     

Responses of group IV afferent units from skeletal muscle to stretch,contraction and chemical stimulation

Summary In an attempt to differentiate between nociceptive group IV muscle receptors and ergoceptive ones, the discharges of single group IV fibres from skeletal muscle in response to local pressure, sustained stretch, repetitive contraction and intra-arterial injections of bradykinin, 5-hydroxytryptamine (5-HT), potassium, phosphate, and lactate were studied in anaesthetized cats.Of the 75 fibres of the study, 5 units were activated by sustained stretch, the responses occurring with a delay. These stretch-sensitive units could not be activated by local pressure or muscular contraction. Thirteen group IV afferents raised their discharge frequency during repetitive contractions. Some of the units responded immediately with the onset of the contractions, whereas the others showed a pronounced delay.Forty-six units were tested with all or most of the above mechanical and chemical stimuli. In 32 afferents a response to at least one of the stimuli was present. Taking only these units into account, several groups of receptors could be distinguished by their different response combinations. One group was activated by pain-producing substances, but not by muscular activity and thus showed nociceptive properties. Another group showed a raised activity during muscular contractions but did not respond to the algesic agents bradykinin and 5-HT. Units belonging to this group might serve as ergoceptors. The borderline between the two groups was not sharp, a considerable number of group IV afferents was found which had both nociceptive and ergoceptive properties.     

Evaluation of a communication and stress management training programme for infertile couples

OBJECTIVE: This study evaluates a patient education programme focussed on improving communication and stress management skills among couples in fertility treatment. METHODS: In total, 37 couples completed the intervention. Two teachers conducted all the five courses offered. The effectiveness regarding communication and infertility-related stress was assessed by questionnaires immediately before (time T1) and after the intervention (time T2). Seeking of information and professional support was assessed at a 12-month follow-up (time T3); response rates were: T1, 93.2%; T2, 85.1%; T3, 74.3%. Data were compared at baseline (T1) and at the 12-month follow-up (T3) with a prospective cohort of Danish people in fertility treatment. RESULTS: There were no differences in infertility-related stress at base line between the two groups studied. We estimated the bi-directional changes in communication, e.g., changes from talking often to talking less frequently and vice versa. More intervention participants started to talk often with their partner about infertility and its treatment after the intervention compared to those who stopped to talk often. Women and men changed occurrence, frequency and content of communication with close other people. Among women marital benefit increased significantly. Infertility-related stress was not reduced significantly. Significantly more intervention participants than in the comparison group had contacted support groups, a psychologist and/or agencies for adoption at the 12-month follow-up. CONCLUSION: The intervention resulted in important perceived improvement in the participants' competence to actively manage changes in marital communication and in communication in different social arenas. PRACTICE IMPLICATIONS: We recommend fertility clinics to develop and evaluate different interventions for those fertility couples who ask for more psychosocial support.     

Diagnosis of rhabdomyosarcomas with HHF35, a monoclonal antibody directed against muscle actins.

The authors have recently developed a monoclonal antibody, HHF35, that recognizes the muscle-specific isoforms of actin. To determine its potential usefulness in the differential diagnosis of "small, round, blue cell" tumors of childhood, they immunolabeled formalinor B-5-fixed tissue sections from known cases of rhabdomyosarcoma or rhabdomyoma (30), neuroblastoma (9), retinoblastoma (2), and Ewing's sarcoma (9) with HHF35 and with antibodies to creatine kinase M, myoglobin, vimentin, and neuron-specific enolase. HHF35 reacted with 29 of 30 cases of rhabdomyosarcoma, whereas antibodies to creatine kinase M and myoglobin were positive on only 12 and 7 tumors, respectively. HHF35 did not react with any case of neuroblastoma, retinoblastoma, or Ewing's sarcoma when the antibody diluent contained 50 mM EDTA. These results indicate that HHF35 is a highly sensitive and specific marker for myogenic differentiation and that it will be useful in the differential diagnosis of rhabdomyosarcomas.     

Differential expression of the HLA-B7 and the HLA-A2 gene in transfected mouse L(tk-) cells after stimulation by mouse interferon

Mouse L(tk-) cells were transfected with recombinant genomic clones encoding the human major histocompatibility antigens HLA-A2 or HLA-B7. The exposure of 15 different transfected cell clones to mouse interferon resulted in an up to 2.9-fold enhancement of the HLA-A2 antigen at the cell surface but in an up to 5.5-fold enhancement of the HLA-B7 antigen as shown by quantitative radioimmunoassay with monoclonal antibodies directed against different HLA epitopes. Using the HLA-Bw6 specific monoclonal antibody 2BC4, an even higher increase of the HLA-B7 antigen (up to 12-fold) could be observed. This higher inducibility of an HLA-B versus HLA-A locus gene may reflect distinct regulatory mechanism controlling the expression of HLA class I subregion antigens.     

Characterization of pili associated with Escherichia coli O18ac.

A strain of Escherichia coli O18ac isolated from the stool sample of a patient with diarrhea was found to agglutinate human erythrocytes. From the results presented it is suggested that this hemagglutination is mediated by pili. Isolated pilus preparations agglutinated human erythrocytes, whereas pilus-negative mutants did not. The serological and chemical analyses indicate that the pili associated with E. coli O18ac are distinct from other types found with E. coli.     

Characterisation of recombinant human IgE-Fc fragments expressed in baculovirus-infected insect cells

IgE mediates its effector functions through the Fc region and it has been demonstrated that structures in the Cvarepsilon3-domain are crucial for FcvarepsilonR-binding. In order to further study structures of importance for the function of IgE, such as the carbohydrates, fragments with unmodified amino acid sequence were blunt-end cloned and expressed in baculovirus-infected Sf9 cells. Two fragments of human IgE, one encompassing the entire Fc-region (rCvarepsilon2-4) and a smaller one comprising the second and third domain (rCvarepsilon2-3), were produced and characterised with respect to epitope expression, glycosylation and FcvarepsilonR-binding. N-terminal analysis showed the expected VCSRDF-sequence of the Cvarepsilon2-domain, confirming correct cleavage of the secretion signal. Immunoblotting and gel permeation chromatography demonstrated that rCvarepsilon2-4 mainly formed a dimer, whereas rCvarepsilon2-3 also existed as monomers and oligomers. Endoglycosidase-treatment revealed that both fragments were N-glycosylated. In inhibition ELISA, rCvarepsilon2-4 and myeloma protein IgE(DES) reacted in a near equimolar way with monoclonal antibodies against the Cvarepsilon2-, Cvarepsilon3- and Cvarepsilon4-domains, whereas rCvarepsilon2-3 only reacted with anti-Cvarepsilon2 mAbs. Moreover, in FACS analysis rCvarepsilon2-4 interacted with two cell-lines constitutively expressing FcvarepsilonRI or FcvarepsilonRII, whereas rCvarepsilon2-3 lacked reactivity. A substantial reduction in the ability of rCvarepsilon2-4, following endoglycosidase treatment, to react with recombinant alpha-chain of the high affinity receptor for IgE in sandwich ELISA, indicated a role of N-linked oligosaccharides in stabilising receptor binding structures. Taken together, our results show that rCvarepsilon2-4, but not rCvarepsilon2-3, will be useful in studies of structure-function relationships of IgE, including the role of N-glycosylation, since it demonstrated appropriate epitope expression, conformation and ability to bind Fcvarepsilon-receptors.     

Delayed responses to electrical stimuli reflect C-fiber responsiveness in human microneurography

The slowing of impulse conduction during the relative refractory period has often been used to assess activation of C-fibers, in particular, in human microneurography. This study aimed to evaluate the sensitivity of this method and the factors affecting it. Thirty cutaneous C-fibers were recorded from the peroneal nerves of healthy human subjects. Intracutaneous electrical stimulation in the receptive field at 4 s intervals, after some minutes of adaptation, induced spike discharges at constant latency. One or more conditioning stimulus pulses were interpolated at different intervals and the increase in latency after the subsequent regular pulse was assessed. The latency shift was found to depend on the number of interposed pulses, on the time interval between conditioning and conditioned stimulus, and on the conduction velocity of the C-unit. The increase in latency was larger with greater distance between stimulating and recording electrodes, indicating a contribution of the conductile membrane over its whole length. On the other hand, slowing was more pronounced, on average, in slower conducting C-units and conduction velocities were slower when recordings were performed more distally. These findings indicate that the slower terminal nerve branches contribute most to the latency increases. Even a single additional spike in between two regular pulses caused a reliable latency shift of 1.2±0.2 ms (mean ±SEM) and additional pulses lead to an approximately linear latency increase (2 pulses: 2.3±0.3 ms; 4 pulses: 5.9±0.7 ms). In contrast to the number of interposed stimuli, different intervals between interposed and regular stimuli had only a minor impact on the latency shifts. It is concluded that latency shifts are reliable indicators of C-fiber activation, being sensitive enough to detect even single spike responses. Furthermore, latency increases may be used as a relative measure of C-fiber activation, e.g., when comparing responses to stimuli of different strength.