Direct recording of nicotinic responses in presynaptic nerve terminals

Nicotinic acetylcholine receptors are widely expressed in the nervous system, but their functions remain poorly understood. One attractive hypothesis is that the receptors act presynaptically to modulate synaptic transmission. We provide a direct demonstration of presynaptic nicotinic receptors in situ by using whole-cell patch-clamp techniques to record currents in large presynaptic calyces that midbrain neurons form on ciliary neurons. Bath application of nicotine induced inward currents in the calyces capable of generating action potentials that overrode the limited space clamp achievable. The inward currents reversed near 0 mV and showed inward rectification common for neuronal nicotinic receptors. Tetrodotoxin (TTX) blocked the action potentials but not the inward currents. alpha-Bungarotoxin blocked both, consistent with the presynaptic receptors containing alpha7 subunits. Recording from the postsynaptic ciliary neurons during nicotine exposure revealed EPSCs that TTX blocked, presumably by blocking presynaptic action potentials. The postsynaptic cells also displayed bimodal inward currents caused by their own nicotinic receptors; the bimodal currents were not blocked by TTX but were blocked partially by alpha-bungarotoxin and completely by D-tubocurarine. Dye-filling with Lucifer yellow from the recording pipette confirmed the identity of patched structures and showed no dye transfer between calyx and ciliary neuron. When calyces or ciliary neurons were labeled en mass with neurobiotin and biocytin through nerve roots, dye transfer was rarely observed. Thus, electrical synapses were infrequent and unlikely to influence calyx responses. Immunochemical analysis of preganglionic nerve extracts identified receptors that bind alpha-bungarotoxin and contain alpha7 subunits. The results unambiguously document the existence of functional presynaptic nicotinic receptors.     

Cross-language synonyms in the lexicons of bilingual infants: one language or two?

This study tests the widely-cited claim from Volterra & Taeschner (1978), which is reinforced by Clark's PRINCIPLE OF CONTRAST (1987), that young simultaneous bilingual children reject cross-language synonyms in their earliest lexicons. The rejection of translation equivalents is taken by Volterra & Taeschner as support for the idea that the bilingual child possesses a single-language system which includes elements from both languages. We examine first the accuracy of the empirical claim and then its adequacy as support for the argument that bilingual children do not have independent lexical systems in each language. The vocabularies of 27 developing bilinguals were recorded at varying intervals between ages 0;8 and 2;6 using the MacArthur CDI, a standardized parent report form in English and Spanish. The two single-language vocabularies of each bilingual child were compared to determine how many pairs of translation equivalents (TEs) were reported for each child at different stages of development. TEs were observed for all children but one, with an average of 30% of all words coded in the two languages, both at early stages (in vocabularies of 2-12 words) and later (up to 500 words). Thus, Volterra & Taeschner's empirical claim was not upheld. Further, the number of TEs in the bilinguals' two lexicons was shown to be similar to the number of lexical items which co-occurred in the monolingual lexicons of two different children, as observed in 34 random pairings for between-child comparisons. It remains to be shown, therefore, that the bilinguals' lexicons are not composed of two independent systems at a very early age. Furthermore, the results appear to rule out the operation of a strong principle of contrast across languages in early bilingualism.     

Polycyclic aromatic hydrocarbon-DNA adducts in human placenta and modulation by CYP1A1 induction and genotype

This study investigated the relationship in human placenta between polycyclic aromatic hydrocabon (PAH)-DNA adduct levels and two biomarkers of cytochrome P4501A1 (CYP1A1): gene induction evidenced by CYP1A1 mRNA, and a genetic polymorphism, the CYP1A1 MspI RFLP. CYP1A1 codes for an inducible enzyme system that catalyzes the bioactivation of PAHs. Prior research found a high correlation in human lung tissue between CYP1A1 activity and DNA damage from PAHs. The CYP1A1 Mspi RFLP has been linked in some studies to risk of lung cancer. The relationships in human placenta between DNA damage, CYP1A1 activity and genotype have not been well characterized and may be relevant to risks from transplacental PAH exposure. The study cohort consisted of 70 newborns from Krakow, Poland, a city with elevated air pollution, and 90 newborns from nearby Limanowa, an area with lower air pollution but greater indoor coal use. Contrary to results seen previously in lung tissue, CYP1A1 mRNA was not significantly correlated with PAH-DNA adduct levels in the placenta. Smoking (self-reported maternal and infant plasma cotinine) was significantly associated with CYP1A1 mRNA levels (P < 0.01), but not with PAH-DNA adduct levels. Placental PAH-DNA adduct levels were significantly higher in infants with the CYP1A1 MspI restriction site compared with infants without the restriction site (P < 0.01), implicating a genetic factor in inter-individual variation in DNA damage in human placenta. Further studies are needed to determine the relevance of this finding to risk of transplacental carcinogenesis.     

Muscle Rad expression and human metabolism: potential role of the novel Ras-related GTPase in energy expenditure and body composition

Ras associated with diabetes (Rad), a new ras-related GTPase, was recently identified by subtractive cloning as an mRNA in skeletal muscle that is overexpressed in NIDDM. To better understand its metabolic significance, we measured skeletal muscle Rad expression in well-characterized insulin sensitive (IS) and insulin resistant (IR) subjects with normal glucose tolerance and in untreated NIDDM patients. We found no differences in expression of Rad mRNA levels among IS, IR, and NIDDM groups using a ribonuclease protection assay (0.22 +/- 0.06, 0.13 +/- 0.01, and 0.16 +/- 0.02 relative units, respectively; NS) and no differences in Rad protein expression using a specific anti-peptide Rad antibody (1.05 +/- 0.18, 1.14 +/- 0.08, and 1.08 +/- 0.21 units/mg protein, respectively; NS). However, Rad protein levels were positively correlated with BMI (r = 0.43, P = 0.03) and percentage body fat (r = 0.55, P < 0.005), two independent measures of obesity, and negatively correlated with resting metabolic rate (r = 0.49, P = 0.01). In multiple regression analyses, percentage body fat and resting metabolic rate independently accounted for 30 and 10% of individual variability in muscle Rad protein expression. In conclusion, Rad expression in skeletal muscle is not altered as a function of insulin resistance or NIDDM in humans. However, these data, for the first time, implicate a role for Rad in regulating body composition and energy expenditure and provide a framework for studies designed to elucidate Rad's cellular functions.     

The incidence of low-pressure urethra as a function of prolapse-reducing technique in patients with massive pelvic organ prolapse (maximum descent at all vaginal sites)

OBJECTIVE: Our aims were to compare several prolapse-reducing techniques during urodynamic evaluation and prospectively evaluate their usefulness in identifying the incidence of low urethral closure pressure in continent patients with massive prolapse. STUDY DESIGN: This preoperative, prospective, repeated-measures urodynamic study evaluated the maximum urethral closure pressure with the use of four different techniques in 30 consecutive continent patients with grade 4 prolapse at all vaginal sites. Twenty patients with grade 0 prolapse served as the control group. All patients from the prolapse group underwent surgical treatment and were followed up clinically for a minimum of 1 year. RESULTS: Use of the Scopette (Birchwood Laboratories, Eden Prairie, Minn.) reduction technique to reduce the prolapse in a linear orientation during multichannel urodynamics revealed a 56% incidence of low-pressure urethra and an overall genuine stress urinary incontinence of 83% in patients with massive pelvic organ prolapse but without clinical urinary incontinence. CONCLUSIONS: There may be an increased indication for sling urethropexy in patients with massive prolapse.     

Paediatric upper gastrointestinal endoscopy: a 2-year review

OBJECTIVE: To review the paediatric upper gastrointestinal endoscopy service in a paediatric department in Hong Kong. METHODS: Records of all endoscopies undertaken in this department from May 1995 to January 1996 were retrieved and analysed. RESULTS: The commonest indication for upper endoscopy was dyspepsia with 88% positive histological findings. Helicobacter pylori (H. pylori) was found in 25% of this group of children. CONCLUSION: Paediatric upper gastrointestinal endoscopy service is invaluable in a paediatric department by providing useful diagnostic information which would otherwise be missed.     

Detection of oxidative stress in heart by estimating the dinitrophenylhydrazine derivative of malonaldehyde

Accurate estimation of the oxidative stress in heart is necessary because the pathogenesis of many heart diseases are believed to be mediated at least in part from the development of oxidative stress resulting from the generation of oxygen free radicals and reduced antioxidant defense system. The most widely used method for this purpose has been the estimation of malonaldehyde (MDA), a lipid peroxidation product, by the thiobarbituric acid (TBA) reaction method. However, because of the nonspecificity of this method, the results are often erroneous. The present report describes a method using high-performance liquid chromatography (HPLC) to estimate MDA. To develop the oxidative stress, two different models were used: ischaemic-reperfused heart and perfusing the heart with a hydroxyl radical (OH+) generating system. The coronary effluents obtained from the isolated rat heart before ischaemia and during the reperfusion of ischaemic heart, as well as during the perfusion of the heart with the OH+ generating system were collected, derivatized with 2,4-dinitrophenylhydrazine (DNPH) and extracted with pentane. Aliquots of 25 microliters in acetonitrile were injected onto a Beckman Ultrasphere C18 (3 microns) column. The products were eluted isocratically with a mobile phase containing acetonitrile-water-acetic acid (40:60:0.1, v/v/v), measured at 307 nm using a Waters M-490 multichannel UV detector and collected for gas chromatography-mass spectrometry (GC-MS). The peaks were identified by co-chromatography with DNPH derivatives of authentic standards, peak addition, and by GC-MS. The retention time for MDA-DNPH was 5.3 min.(ABSTRACT TRUNCATED AT 250 WORDS)     

Stroke therapy in children

OBJECTIVE: To evaluate the treatment modalities used in children (ages 1-18 years) with cerebral infarction. BACKGROUND: [corrected] Cerebrovascular disease in children is more common than once suspected but its treatment has not been rigorously studied. MATERIAL AND METHODS: We reviewed all cases of cerebral infarction at the James Whitcomb Riley Hospital for Children at the Indiana University Medical Center from 01.01.80 to 31.12.95. RESULTS: Ninety-three children who experienced ischemic strokes were followed over the past fifteen years. Fifty-seven males and thirty-six females comprised the sample. Mean age was 6.9 years at the time of stroke. No medication or surgical intervention was the therapeutic recommendation in 44% of patients. For cardioembolic strokes, warfarin was used later in the course for a few patients who went on to have atrial fibrillation or valve replacement. Aspirin was used in all patients with carotid artery dissections. Aspirin was used in most children with Moya-Moya, with calcium channel blockers and surgical intervention used in later cases. Exchange transfusion followed by monthly transfusion and chelation therapy has been the treatment of choice for children with cerebral infarction complicating sickle cell disease. CONCLUSIONS: In most instances, treatment was widely disparate, probably reflecting the lack of firm therapeutic guidelines for this age group, with a better understanding of the etiology and pathophysiology of strokes in children, multicenter, international, randomized therapeutic trials based strictly on an etiological basis should be organized in the future.