Environmental factors in the development of autism spectrum disorders.

Autism spectrum disorders (ASD) are highly heterogeneous developmental conditions characterized by deficits in social interaction, verbal and nonverbal communication, and obsessive/stereotyped patterns of behavior and repetitive movements. Social interaction impairments are the most characteristic deficits in ASD. There is also evidence of impoverished language and empathy, a profound inability to use standard nonverbal behaviors (eye contact, affective expression) to regulate social interactions with others, difficulties in showing empathy, failure to share enjoyment, interests and achievements with others, and a lack of social and emotional reciprocity. In developed countries, it is now reported that 1%-1.5% of children have ASD, and in the US 2015 CDC reports that approximately one in 45 children suffer from ASD. Despite the intense research focus on ASD in the last decade, the underlying etiology remains unknown. Genetic research involving twins and family studies strongly supports a significant contribution of environmental factors in addition to genetic factors in ASD etiology. A comprehensive literature search has implicated several environmental factors associated with the development of ASD. These include pesticides, phthalates, polychlorinated biphenyls, solvents, air pollutants, fragrances, glyphosate and heavy metals, especially aluminum used in vaccines as adjuvant. Importantly, the majority of these toxicants are some of the most common ingredients in cosmetics and herbicides to which almost all of us are regularly exposed to in the form of fragrances, face makeup, cologne, air fresheners, food flavors, detergents, insecticides and herbicides. In this review we describe various scientific data to show the role of environmental factors in ASD.

Detection of anthrax toxin genetic sequences by the solid phase oligo-probes.

PURPOSE: There is an urgent need to detect a rapid field-based test to detect anthrax. We have developed a rapid, highly sensitive DNA-based method to detect the anthrax toxin lethal factor gene located in pXO1, which is necessary for the pathogenicity of Bacillus anthracis. MATERIALS AND METHODS: We have adopted the enzyme-linked immunosorbent assay (ELISA) so that instead of capturing antibodies we capture the DNA of the target sequence by a rapid oligo-based hybridization and then detect the captured DNA with another oligoprobe that binds to a different motif of the captured DNA sequences at a dissimilar location. We chose anthrax lethal factor endopeptidase sequences located in pXO1 and used complementary oligoprobe, conjugated with biotin, to detect the captured anthrax specific sequence by the streptavidin-peroxidase-based colorimetric assay. RESULT: Our system can detect picomoles (pMoles) of anthrax (approximately 33 spores of anthrax) and is >1000 times more sensitive than the current ELISA, which has a detection range of 0.1 to 1.0 ng/mL. False positive results can be minimized when various parameters and the colour development steps are optimized. CONCLUSION: Our results suggest that this assay can be adapted for the rapid detection of minuscule amounts of the anthrax spores that are aerosolized in the case of a bioterrorism attack. This detection system does not require polymerase chain reaction (PCR) step and can be more specific than the antibody method. This method can also detect genetically engineered anthrax. Since, the antibody method is so specific to the protein epitope that bioengineered versions of anthrax may not be detected.

Orlistat maintains biliary lipid composition and hepatobiliary function in obese subjects undergoing moderate weight loss.

OBJECTIVES: Orlistat, an intestinal lipase inhibitor, has recently been approved by the US Food and Drug Administration for treatment of obesity. The effects of orlistat on hepatobiliary function have not been previously defined. A 4 wk study was performed involving modest weight loss in obese subjects to observe any short-term hepatobiliary responses that occur after initiating treatment with orlistat and a hypocaloric diet. METHODS: A total of 23 obese (BMI 30-41 kg/m2) subjects were randomized to a double blind t.i.d. treatment with 120 mg of orlistat or a placebo in conjunction with a hypocaloric diet (1200-1500 kcal/day). The study was designed to achieve similar modest weight loss in both groups in order to be able to directly assess the effects of orlistat. Cholesterol saturation, bile composition, and gallbladder motility were measured. RESULTS: At the end of the treatment period, mean weight loss of 3.8 kg was achieved in the orlistat group (vs 2.3 kg with placebo, p = NS). Total bile acid concentration decreased significantly with placebo (-18.57 +/- 6.99 mmol/L; 95% CI = -32.26 to -4.87), but not with orlistat. Biliary phospholipid concentration decreased significantly with placebo (-4.38 +/- 1.91 mmol/L; 95% CI = -8.13 to -0.64) but not with orlistat. Mean changes from the baseline in cholesterol saturation index and gallbladder motility were similar in both groups. Microscopy of bile failed to reveal cholesterol microcrystals before or after treatment in either group. CONCLUSIONS: Our findings indicate a primary initial effect of weight loss is a reduction in biliary bile acids and phospholipids. Orlistat blocks these adverse changes in biliary lipid composition and maintains hepatobiliary function. We speculate that the risk of formation of gallstones during weight loss may actually be lowered with orlistat.

Short-term orlistat treatment does not affect mineral balance and bone turnover in obese men.

Orlistat is a gastrointestinal lipase inhibitor that is used to reduce dietary fat absorption and to enhance weight loss in subjects consuming a hypocaloric diet. To assess whether orlistat has an effect on the metabolism of six minerals, a 21-d, double-blind, randomized, parallel-group, placebo-controlled mineral balance study was conducted in obese (body mass index > 30 kg/m(2)) men. Subjects consumed a hypocaloric diet with a constant daily mineral content and received daily oral treatment with orlistat (120 mg three times daily) (n = 14) or placebo (three times daily) (n = 14) for 21 d. After a 14-d equilibration period, calcium, phosphorus, magnesium, iron, copper and zinc balances were assessed for d 15-21. In addition, the effect of diet and orlistat treatment on bone metabolism was estimated from measurement of biomarkers of bone formation and bone resorption. Serum and urine electrolytes were also measured at baseline and at the end of treatment. Orlistat inhibited fat absorption by approximately 33% (P < 0.05). There were no significant differences in mineral apparent absorption, urinary mineral loss or mineral balance between the orlistat and placebo groups. Markers of bone turnover and serum and urine electrolytes did not differ between the orlistat and placebo groups. Orlistat was well tolerated; adverse events were of mild or moderate intensity, and the majority of these events were unrelated or remotely related to study treatment. In obese men consuming a hypocaloric diet, the administration of orlistat had no significant effect on the balance of six selected minerals. In addition, biomarkers of bone turnover, as well as serum and urine electrolytes, were not affected by orlistat treatment.

A new mixed micellar preparation for oral vitamin K prophylaxis: randomised controlled comparison with an intramuscular formulation in breast fed infants.

OBJECTIVE: To compare a new oral preparation of vitamin K1 (Konakion MM) containing lecithin and glycocholic acid with a standard intramuscular (IM) preparation during the first 8 weeks of life in exclusively breast fed infants. METHODS: Infants were randomised at birth to the IM group (1 mg vitamin K) or the oral group (2 mg given at birth and repeated at 7 and 30 days of life). Prothrombin time (INR), plasma vitamin K1, and PIVKA II (undercarboxylated prothrombin) were monitored at 14, 30, and 56 days of age. RESULTS: Seventy nine infants were randomised to the oral group and 77 to the IM group. Sixty seven infants in each group completed eight weeks of the study. Prothrombin times did not differ between the two groups. Mean (SD) plasma vitamin K1 values (in ng/ml) decreased in both groups over time, but were higher in the oral group at 14 and 56 days: 2.0 (1.6) v 1.3 (1.1) at 14 days; 0.5 (0.3) v 0.5 (0.7) at 30 days; and 0.5 (0.8) v 0.2 (0.2) at 56 days of life. PIVKA II was raised (> or = 0.1 AU/ml) in cord blood in 47% of the infants. By 14 days, only one infant in each group had a raised PIVKA II value and both of these initially had high concentrations of PIVKA II in cord blood. At 30 days, there were no raised PIVKA II values. At 56 days, there were no raised PIVKA II values in the oral group, although three infants in the IM group had raised values. CONCLUSIONS: Plasma vitamin K concentrations were at least equal or significantly higher in babies given oral vitamin K supplements compared with IM treated babies at the time points measured. Through the first 8 weeks of life, multiple doses of the new oral preparation maintain haemostasis and vitamin K status in breast fed infants at least equal to that of the intramuscular preparation.

Evaluation of methods of administering tyramine to raise systolic blood pressure.

To compare the relative merits of two different administration regimens, tyramine was administered intravenously in ascending doses to 12 healthy subjects to raise systolic blood pressure slightly more than 30 mm Hg. Six subjects received tyramine by bolus injection and six other subjects received tyramine by infusion. The bolus dose of tyramine needed was 4.34 +/- 1.51 mg (X +/- SD) and the infusion rate needed was 1.11 +/- 0.33 mg/min. Four blood pressure response patterns to continuous tyramine infusion were observed. Because different units were measured for the quantity of tyramine administered, the between-subject variance estimate to within-subject variance estimate ratios were calculated. The two techniques had equivalent consistency. With the bolus method, in contrast to the infusion procedure, the dose-response relationship was obvious in most subjects. Therefore the bolus method was judged to be more useful than the infusion method.

Inhibitory control of proximal colonic motility by the sympathetic nervous system.

The purpose of this study is to determine whether or not the sympathetic nervous system provides a tonic inhibitory input to the colon in chloralose-anesthetized cats. Proximal and midcolonic motility were monitored using extraluminal force transducers. An intravenous bolus injection of 5 mg of phentolamine in 14 animals elicited a pronounced increase in proximal colon contractility. The minute motility index changed from 0 +/- 0 to 26 +/- 4 after phentolamine administration. Midcolonic motility also increased in response to phentolamine. Specific blockade of alpha 2-receptors, but not alpha 1-receptors, caused the same response seen with phentolamine. alpha-Adrenergic blockade increased colon contractility after spinal cord transection but not after ganglionic blockade. Blockade of alpha-adrenergic receptors was also performed before vagal and pelvic nerve stimulation and in both cases increased colonic motility. Vagal stimulation alone had no effect on colonic contractility, while pelvic nerve stimulation increased motility at the midcolon. alpha-Receptor blockade did not alter the ineffectiveness of vagal stimulation but did unmask excitatory effects of pelvic nerve stimulation on the proximal colon. All excitatory colonic responses were prevented by blocking muscarinic cholinergic receptors. These data indicate that tonic sympathetic nervous system activity exerts an inhibitory effect on colonic motility. The inhibitory effect is mediated through alpha 2-adrenergic receptors. Based on these findings, we suggest that alterations in sympathetic nervous system activity may be extremely important for the regulation of circular muscle contractions in the colon.

Effect of brief bursts of carotid sinus stimuli on heart rate and atrioventricular conduction.

The effect of a single brief stimulus burst applied simultaneously to both carotid sinus nerves on atrioventricular conduction (PR) was examined in paced and unpaced preparations of anesthetized open-chest dogs. The relative timing of the stimulus burst was varied to encompass the complete cardiac cycle. Carotid sinus/vagal effect curves were constructed to identify the time course of the response. In paced preparations the maximum increase in PR was 20.3 +/- 2.7 msec and this occurred 458.0 +/- 22.8 msec after the stimulus. There was a latency of 246.0 +/- 12.4 msec after the electrical stimulus before the PR began to increase. In unpaced heart preparations the effect of single carotid sinus nerve stimuli on heart period (PP) and PR was also determined. PP was maximally lengthened by 251.4 +/- 49.4 msec at 644.2 +/- 50.9 msec after the stimulus. There was a latency of 251.4 +/- 10.5 msec before the first noticeable change in PP occurred. The PR response was biphasic. The maximum lengthening of the PR interval was 16.2 +/- 4.2 msec. This response occurred at 361 +/- 22.0 msec after the electrical stimulus. The PR decreased to a minimum value of 13.6 +/- 2.2 msec below control values at 767.1 +/- 44.0 msec after the stimulus. The overall effect of carotid sinus activity on atrioventricular conduction depends not only on the direct nodal effect of acetylcholine but also on the indirect heart rate changes. We conclude that brief bursts of carotid sinus nerve stimulation produce cardiac electrophysiological effects qualitatively similar to the effects of direct vagal stimulation, differing only in having a lower amplitude, a longer latency and a somewhat wider time dispersion.

Effects of digoxin on the chronotropic responses to repetitive vagal stimulus bursts in the dog.

We studied the effects of digoxin on the chronotropic responses of the heart to repetitive bursts of vagal stimulation in chloralose-anesthetized dogs. The frequency of the stimulus bursts was increased linearly with time. Over a certain range of frequencies, the cardiac pacemaker became synchronized with the vagal stimulation in a 1:1 ratio of heart beats to stimulus bursts. Digoxin increased the range of cardiac cycle lengths over which 1:1 synchronization occurred during repetitive vagal stimulation. This increment in the range of synchronization varied directly with the dose of digoxin. Before digoxin was given, the range of cardiac cycle lengths over which synchronization occurred when the vagus nerve was stimulated with 10 pulses per burst was 272 +/- 50 (mean +/- SE) ms. However, after a cumulative dose of 120 micrograms/kg-1 digoxin had been given, the range of 1:1 synchronization increased to 396 +/- 32 ms. Digoxin did not appear to have a proportionately greater effect on those processes that take place in the phase of the cardiac cycle during which the pacemaker cells are maximally responsive than on those processes that occur in the phase of the cycle during which the pacemaker cells are minimally responsive. Therefore, we conclude that the augmented entrainment induced by digoxin is ascribable to its tendency to enhance the chronotropic response to vagal stimulation.

The rate of arterial blood pressure change as a factor in the hysteresis of the baroreceptor reflex.

We induced sinusoidal arterial blood pressure oscillations in anaesthetized dogs, and measured the resulting reflex changes in cardiac cycle length. The graphs of cardiac cycle length as a function of systolic arterial pressure displayed considerable hysteresis. The extent of the hysteresis and the direction around the loop varied with the frequency of the pressure oscillations. At relatively low frequencies (0.02 and 0.04 Hz) the loops were predominantly anticlockwise, whereas at higher frequencies (0.08 and 0.16 Hz) the loops were mainly clockwise in direction. In a second group of dogs we stimulated the transected vagus nerves with sequential ramp-like increases and decreases in voltage. The graph of the chronotropic response displayed a prominent anticlockwise hysteresis. Hence the anticlockwise hysteresis which was induced reflexly by cyclic oscillations of systemic arterial pressure is at least partly ascribable to an efferent vagal mechanism.

Improvement of the toxic to inotropic dose ratio of digoxin by excluding central nervous system influences on the heart.

The effect of progressive digoxin administration (20 micrograms/kg/15 min, i.v.) on cardiac contractile force, cardiac output, heart rate, arterial blood pressure and total peripheral resistance was examined in chloralose anesthetized cats with and without central nervous system control of the heart. In neurally intact animals, the inotropic response (i.e., a sustained increase of 0.5 g-tension in contractile force) occurred with 83 +/- 22.1 micrograms/kg digoxin. The peak change in force occurred immediately prior to the development of ventricular arrhythmias and attained a value of + 1.88 +/- 0.84 g-tension above the control level. Toxicity (i.e., ventricular arrhythmias) occurred with 158 +/- 8.0 micrograms/kg and death occurred with 188 +/- 8.4 micrograms/kg digoxin. The toxic to inotropic, and lethal to inotropic dose ratios were 3.62 +/- 1.28 and 4.28 +/- 1.50, respectively. Elimination of central nervous system control to the heart by bilateral cervical vagotomy and spinal section at the atlanto-occipital junction significantly (p less than 0.05) improved the toxic to inotropic and lethal to inotropic dose ratios to 6.66 +/- 1.00 and 8.72 +/- 1.29, respectively. In addition, the inotropic response attained in these animals (+ 5.30 +/- 0.70 g-tension above control level) was significantly higher than the inotropic response attained in the neurally intact animals. These results suggest that when the central nervous system actions of digoxin are prevented from influencing the heart, greater increases in cardiac contractility are obtained with less likelihood of digoxin-induced toxicity.

An improved signal processor for the ultrasonic dimension gauge.

The flip-flop producing a pulse proportional to the transit time of an ultrasonic burst between two piezoelectric crystals was often reset by transmission artefacts in sonomicrometers. A design improvement was made. The insertion of two monostable vibrators and one nand gate corrected the resetting caused by transmission artefacts. The new design produced a flip-flop pulse that was proportional to the transit time of the ultrasonic burst through the aorta. Ascending aortic diameter was monitored in unanaesthetised dogs for 6 to 8 weeks using the new design.

Digoxin-induced decrease in intraocular pressure in the cat.

A constant intravenous infusion of digoxin (2 micrograms/kg/min) to alpha-chloralose-anesthetized cats produced a progressive decrease in intraocular pressure with increasing doses of digoxin between 60 micrograms/kg and drug-induced ventricular arrhythmia which occurred at a mean dose of 172 micrograms/kg. Digoxin elicited a 40% decrease in intraocular pressure just prior to ventricular arrhythmia compared to a decrease of only 10% with a control infusion of diluent in the same animal (P less than 0.05). There was no significant difference (P less than 0.1) between changes in blood pressure and heart rate observed in the experimental versus the control infusions. The decrease in intraocular pressure may result from inhibition of the Na-K-ATPase in the ciliary body.

Interactions between digoxin and brief vagal bursts influencing atrioventricular conduction.

The interaction between intravenous injections of digoxin (20 microgram/kg every 15 minutes) and brief electrical bursts of vagal stimulation was determined in chloralose-anesthetized dogs. Vagal effect curves were generated to characterize the effect of brief vagal stimulus bursts on atrioventricular conduction. These curves were fit with an analytic expression from which the following parameters were derived as the experimental observations: 1) the maximal change in atrioventricular conduction (deltaPRmax), 2) the time after the stimulus at which atrioventricular conduction was maximally inhibited (Tmax) and 3) the width of the vagal effect curve at one-half the maximal amplitude (TD). Digoxin administration significantly (P less than .05) increased deltaPRmax, Tmax and TD by 21.6 +/- 4.3, 50.0 +/- 16.1 and 125.5 +/- 42.4 msec, respectively, before the disruption of sinus rhythm. Diluent or saline administration did not alter deltaPRmax, Tmax or TD. In addition, digoxin produced dose-dependent increases in deltaPRmax. These results suggest that digoxin vagal interactions not only affect the PR interval but also the time to the maximum delay in atrioventricular conduction and the length of time for depressed conduction after brief bursts of vagal activity.

Neuroexcitatory effects of digoxin in the cat.

The effect of intravenous injections of digoxin (20 mug/kg every 15 minutes) on spontaneously occurring activity in autonomic efferent nerves, motor nerves, afferent nerves, electrocardiogram and on arterial blood pressure was evaluated in chloralose-anesthetized cats. Administration of digoxin enhanced neural activity in pre- and postganglionic cardiac synpathetic nerves and this enhancement occurred near the time the disturbances in ventricular rhym were noted. Neural activity continued to increase during ventricular tachycardia and maximum enhancement was observed just proir to ventricular fibrillation. Similar results were observed when digoxin was administered to animals in which neural activity was recorded from preganglionic splanchnic and superior cervical nerves. Digoxin administration also increased discharge frequency from vagus (efferent fibers), phrenic and carotid sinus nerves. Denervation of cardiovascular reflexogenic areas prevented the increased discharge in vagus nerves, reduced it in phrenic nerves, but did not affect nerve discharge in sympathetic nerves. These results suggest that digoxin-induced hyperactivity in synpathetic nerves was related to a central nervous system effect of the drug, whereas the mechanism for the digoxin-induced hyperactivity in vagus nerves involved a peripheral reflex effect of the drug. Both sites were involved in the digoxin-induced hyperactivity in phrenic nerves. Enhancement of cardiac sympathetic nerve activity appeared to be responsible for the ventricular arrhythmias provoked by digoxin as 1) a temporal relationship was observed between augmented nerve activity and arrhythmia development, 2) a centrally acting sympathetic nervous system depressant drug, clonidine, converted the ventricular arrhythmia to normal rhythm, and 3) removal of sympathetic influence to the heart by spinal cord transection decreased the sensitivity of the heart to the arrhythmogenic effect of digoxin. These results suggest that digoxin partially responsible for its cardiotoxic effects.

Role of the nervous system in experimentally induced arrhythmias.

The purpose of our studies was to examine the role of the nervous system in arrhythmias produced by digitalis overdose and coronary artery occlusion in the cat. This was done by observing the effect of these arrhythmogenic procedures on cardiac efferent neural activity and then determining whether any observed alteration in neural activity contributed to the cardiac rhythm disturbances evoked by digitalis and coronary artery occlusion. Our data indicate that both procedures used to evoke arrhythmias activate each division of the autonomic nervous system. Activation of the sympathetic nervous system resulted in a deleterious effect on cardiac rhythm whereas activation of the parasympathetic nervous system, in general, resulted in a beneficial effect on cardiac rhythm. With coronary occlusion, the role exerted by the nervous system depended on the anatomic location of the involved myocardium. Studies directed at elucidating the mechanisms whereby the nervous system caused cardiac rhythm disturbances indicated that there may be an important difference between the antiarrhythmic efficacy of beta-adrenergic blockade and bilateral stellate ganglionectomy. The latter procedure proved to be a more effective way of removing deleterious sympathetic neural effects on the heart. In conclusion, our findings suggest that the development of new drugs for treating arrhythmias resulting from digitalis and coronary occlusion should be aimed at finding drugs that act to either depress central sympathetic outflow or enhance parasympathetic effects on the ventricle.