Regulation of pharyngeal motility by FMRFamide and related peptides in the medicinal leech, Hirudo medicinalis.

The medicinal leech possesses FMRFamide-like immunoreactivity in neural processes and somata associated with the pharynx and pharyngeal ganglia. The pharynx possessed about 25 immunoreactive somata; about half of the approximately 20 neurons of the pharyngeal ganglia were immunoreactive. We provide brief descriptions of several neurons located in the first neuromere of the subesophageal ganglion involved in controlling pharyngeal motility. Double-labeling experiments indicate that one of these cells, named Swallow neuron 1 (SW1), contains a FMRFamide-like peptide. Stimulation of SW1 caused the mouth to open and the pharynx to dilate. Upon termination of SW1 stimulation, the mouth closed, and a peristaltic wave progressed from the mouth down the length of the pharynx. Stimulation of SW1 did not produce 1:1 postsynaptic potentials in pharyngeal muscle cells. Thus, SW1 is apparently not a motor neuron. The pharynx responded to application of FMRFamide and related peptides by producing a series of 20- to 35-s phasic contractions superimposed upon an increase in basal tonus. The peptide-induced response was quantified by measuring increases in basal tonus, peak tension, and integrated area. Although there were some differences in the order of potency depending upon which parameter was considered, the approximate order of potency of RFamide peptides tested was: pQDPFLRFamide > or = FMRFamide approximately YGGFMRFamide > or = YMRFamide approximately FLRFamide approximately GGKYMRFamide approximately YLRFamide > leucomyosuppressin approximately perisulfakinin. Except for differences in potency, each of the RFamide peptides produced similar contractile waveforms. FMRFamide-induced responses were reduced by the protein kinase C inhibitor bisindolylmaleimide I (10 microM), the nonspecific protein kinase inhibitor H-7 (50 microM), and were increased by the protein phosphatase inhibitor okadaic acid (1 microM). However, the FMRFamide-induced response was unaffected by the protein kinase A inhibitor H-89 (1 microM), the phosphodiesterase inhibitor theophylline (1 mM), the phospholipase A(2) inhibitor OBAA (0.1 microM) or the cation channel blocker amiloride (100 microM).

[Detection of varicella zoster virus infections using polymerase chain reaction]. / Nachweis von Varizella-Zoster-Virus-Infektionen mittels Polymerasekettenreaktion.

The polymerase chain reaction (PCR) was used to detect varicella zoster virus (VZV) DNA in vesicle samples from patients with varicella and zoster. Primers and the oligonucleotide probe were chosen from the region of the immediate early gene 63. Procedures for preparing the DNA from the specimens were omitted, and the amplified DNA was directly detected in ethidium bromide-stained polyacrylamide or agarose gels, thus providing a rapid and less laborious assay. A total of 66 vesicle specimens including 3 crusts (collected on days 1-14 after the onset of exanthem) were tested by the simplified VZV-PCR, and 64 (97%) were positive. When the direct visualization of the amplified DNA was confirmed by DNA hybridization, a non-radioactive hybridization assay involving a digoxigenin-labelled oligonucleotide probe and detection by chemiluminescence proved as adequate as a radioactive hybridization assay. Thus, the VZV PCR described appears to be a useful diagnostic test for detecting and identifying varicella zoster virus.

Diagnosis of acute and latent varicella-zoster virus infections using the polymerase chain reaction.

A simplified assay for the diagnosis of varicella-zoster virus (VZV) infections based on the polymerase chain reaction (PCR) is described. Omitting the procedures for extraction and purification of DNA, the crude vesicle fluid materials were used for PCR. Moreover, hybridization was not necessary for detection of the amplification products because they were already visible after ethidium bromide staining of the electrophoresis gel. Results could therefore be obtained within one day. In comparison to virus isolation, PCR proved much more rapid, highly sensitive, and specific. DNA extraction and a double PCR assay with nested primers were necessary for detection of latent VZV infections in trigeminal and thoracic ganglia. The data suggest that the procedures described are universally applicable to several types of specimens dependent on the calculated amount of target DNA.