Sensitivity of a ribosomal RNA gene probe for identification of life stages of Anopheles arabiensis and An. gambiae (Diptera: Culicidae) using three storage methods.

Individual larvae, pupae, female adults, and adult body parts of Anopheles arabiensis Patton and An. gambiae Giles were stored for 1 mo either in isopropanol at room temperature, over a desiccant at room temperature, or at -70 degrees C. DNA was extracted, digested with EcoR1 restriction enzyme, subjected to electrophoresis in agarose gel, transferred to filters, then hybridized to a 32P-labeled rDNA probe. There was no difference among storage treatments in the proportion of correctly identified samples. First instars were not identifiable. Pupae and female adults were more likely to be identified than earlier life history stages. Nonetheless, the probe identified greater than 75% of second instars, 94% of third instars, and 74% of fourth instars. There were no differences between the species in the proportion of identifiable samples for any life history stage.     

Unmyelinated cutaneous afferent neurons activate two types of excitatory amino acid receptor in the spinal cord of Xenopus laevis embryos

The trunk and tail skin of Xenopus laevis embryos near the time of hatching is innervated by the mechanoreceptive free nerve endings of Rohon-Beard neurons, a homogeneous class of cutaneous primary afferent fibers. Rohon-Beard neurons have cell bodies and axons in the dorsal spinal cord, where they monosynaptically excite a population of dorsolaterally situated interneurons (Clarke and Roberts, 1984). EPSPs can be recorded in these dorsolateral interneurons following electrical stimulation of the unmyelinated neurites of Rohon-Beard neurons in the skin. The EPSPs are dual component, consisting of separate fast and slow potentials that are usually evoked synchronously and that closely resemble those described previously in Xenopus and lamprey motoneurons (Dale and Roberts, 1985; Dale and Grillner, 1986). The excitation of dorsolateral interneurons by Rohon-Beard neurons is reduced by the bath application of excitatory amino acid antagonists. Kynurenic acid suppresses both the fast and slow components of the EPSPs, while both (+/-)-2-amino-5-phosphonovaleric acid (APV) and 1 mM magnesium reduce the slow component but have little or no effect on the peak amplitude of the EPSPs. These data suggest that Rohon-Beard neurons release an excitatory amino acid neurotransmitter, which acts simultaneously at both N-methyl-D-aspartate (NMDA) and non-NMDA receptor types. This is the first direct demonstration of dual-component excitatory amino acid-mediated synaptic transmission from cutaneous primary afferent neurons in the vertebrate spinal cord. The bath application of the agonists NMDA, kainate, or quisqualate in salines containing 1 microM TTX depolarized the interneurons and reduced their input resistance, which suggests that the interneurons possess all 3 types of excitatory amino acid receptor. Kynurenic acid strongly inhibits responses to NMDA and kainate, but is relatively less effective against the larger responses of quisqualate in this system.     

Inadvertent removal of foreign body via a flexible fiberoptic bronchoscope in a 4-year-old boy

We report the case of a 4-year-old child with an atypical presentation of an aspirated unwitnessed foreign body in the airway. During an attempt to obtain a biopsy of what appeared to be a mucosal growth, the foreign body was removed. This may be the first report of foreign body removal in a child this young, using a flexible fiberoptic bronchoscope. Pediatr Pulmonol. 1994; 18:51–52. © 1994 Wiley-Liss. Inc.     

1,2,4-Triazolo[4,3-a]pyrazine derivatives with human renin inhibitory activity. 1. Synthesis and biological properties of alkyl alcohol and statine derivatives

A series of 1,2,4-triazolo[4,3-a]pyrazine derivatives with human renin inhibitory activity, which incorporate (1S,2S)-2-amino-1,3-dicyclohexyl-1-hydroxypropane, statine (Sta), and (3S,4S)-4-amino-5-cyclohexyl-3-hydroxy-pentanoic acid (ACHPA) transition-state mimetics, have been prepared. Structure-activity relationships for renin inhibitory activity in the series are consistent with the 2-[8-isobutyl-6-phenyl-1,2,4-triazolo[4,3-a]pyrazin-3-yl]-3-(3 pyridyl)propionic acid moiety 10b acting as a non-peptidic replacement for the P4-P2 (Pro-Phe-His) residues of the natural substrate angiotensinogen. Compounds 12m, 12o and 12q were potent inhibitors of partially purified human renin (IC50 values 1.7, 6.8, and 3.7 nM, respectively), and also effectively lowered blood pressure in anesthetized, sodium depleted marmosets following intravenous administration. On oral administration however, no blood pressure lowering activity could be detected, and absorption studies in bile duct cannulated rats indicate that this may be due primarily to poor oral absorption, rather than rapid biliary excretion. The reason for the observed poor oral activity is not clear, but it seems unlikely that poor aqueous solubility or metabolic instability to gut enzymes are rate-determining, and other factors such as high molecular weight may also be very important.     

Anesthesia and/or Sedation for Pulsed Dye Laser Therapy

One of the most exciting developments in pediatric dermatology has been the use of the flashlamp-pumped, 585-nm, pulsed dye laser for treatment of vascular birthmarks. In many cases the results are miraculous. The increase in self-esteem and happiness of many children and adolescents has been overwhelming; for some, depression has been lifted, stuttering has ceased, social involvement has increased, and antidepressants have been discontinued. There are many success stories to tell.
Despite the remarkable effects of the pulsed dye laser and the medical and psychosocial indications for its use, the issue of pain control remains significant. We have no perfect outpatient pediatric anesthetic. Most methods carry either some risk or, if not hazardous, often are not very effective for controlling pain. Needless to say, a diversity of opinions exist on how to manage discomfort from this treatment modality. Therefore, we thought it would be useful to share the experiences and opinions of several dermatologists who have extensive experience with the pulsed dye laser.     

Detection of intra-amniotic infection by gas-liquid chromatography.

The use of gas-liquid chromatography to detect short-chain organic acids in the amniotic fluid of patients with amnionitis has been previously described. Most of the studies describe patients in the early third trimester with such infections. The purpose of the current study was to confirm the correlation of infection with increased production of organic acids and to assess the effect of gestational age on the presence of these short-chain fatty acids in the amniotic fluid. Six patients with confirmed chorioamnionitis were used as positive control subjects. Seventy-two patients at various gestational ages from 18 to 42 weeks with negative Gram's stain and culture results from the amniotic fluid were used as negative control subjects. The data revealed an increased production of organic acids, particularly pyruvic, oxalic and succinic, in patients with amnionitis regardless of gestational age. Interestingly, patients with noninfected amniotic fluid also revealed an increase in the concentrations of volatile organic acids between 27 and 32 weeks' gestation. It appears from this study that previous results correlating chorioamnionitis with an increased production of organic acids in the amniotic fluid may have been confounded by gestational age.     

Brainstem neurons projecting to different levels of the spinal cord of the dogfish Scyliorhinus canicula.

Neurons in the brain that project to different levels of the spinal cord in the dogfish Scyliorhinus canicula have been identified by retrograde labelling with horseradish peroxidase. Injections have been made at four different levels, namely, cervical (segments 3-6), pectoral (segments 16-18), pelvic (segments 34-36) and caudal (segments 60-80). Labelled neurons were located in the diencephalon, mesencephalon and rhombencephalon. Twenty nuclei contained labelled cells following cervical injections. Fourteen of these contained labelled cells following pectoral injections as did nine following pelvic injections and only seven after caudal injections. Of the three diencephalic nuclei projecting to the cord only nucleus thalamus ventralis pars medialis projected further than cervical regions. From the mesencephalon, the tectospinal projection, which arises from tectal and tectotegmental regions, reaches only as far as the cervical cord. The reticulospinal system, arising from cell groups in both the mesencephalon and rhombencephalon, provides the major descending pathway and reaches the most caudal levels. From the octaval region a vestibulospinal pathway arises from nucleus octavus magnocellularis, to reach all levels of the cord, and from nucleus octavus descendens, to project only as far as the pectoral cord. Other rhombencephalic cell groups--the trigeminal nuclei and nuclei B, F and G--project at least as far as the pectoral cord. We conclude that few brainstem nuclei can directly affect the whole spinal cord, whereas many nuclei may have an impact on its most rostral regions; here we have found labelled spinal interneurons with long descending axons that reach the most caudal cord levels.