Potential anti-inflammatory treatments against cutaneous sulfur mustard injury using the mouse ear vesicant model.

In spite of several decades of research, no effective treatment to skin injuries following exposure to sulfur mustard (HD) has yet been found. In the present study, the mouse ear vesicant model was applied to awake mice in order to evaluate the efficiency of potential anti-inflammatory treatments in preventing HD-induced skin damages. Clinical follow-up and histological evaluation were used to characterize the injuries to the skin and to evaluate the efficiency of the drugs that were applied. Thus, the extent of mouse ear oedema and the histopathological changes following a single application of 0.2 or 1 microL of neat HD for 10 min (representing moderate and severe lesions, respectively), were monitored. Typical HD skin lesions were observed including epithelial and dermal damage. The development of the injury in mouse ears was found to be very similar to that reported in human skin. Screening of post-exposure topical steroids and non-steroidal antiinflammatory drugs (NSAIDs) proved that HD-induced inflammation could be diminished significantly as long as the treatment was applied during the early stages following exposure. A combined application of these drugs approved to be particularly effective in reducing inflammation.

Scopolamine bioavailability in combined oral and transdermal delivery.

Transdermal therapeutic system scopolamine (TTS-S) is effective in preventing motion sickness for 72 h. However, by this route a prophylactic effect is obtained 6 to 8 h postapplication. By the oral route, scopolamine is effective within 0.5 h for a period of 6 h. To achieve safe as well as effective protection against seasickness during the first hours of a voyage until the TTS-S patch takes effect, the pharmacokinetics of scopolamine was investigated after patch application in combination with oral tablets, 0.6 mg, 0. 3 mg, or placebo. Subjects were 25 naval-crew volunteers, randomly divided into three groups: group 1 (n = 9), TTS-S patch + 0.6 mg of scopolamine per os (p.o.); group 2 (n = 8), TTS-S patch + 0.3 mg of scopolamine p.o.; and group 3 (n = 8), TTS-S patch + placebo tablet. Blood samples were collected before treatment and 0.5, 1, 1.5, 2.5, 3.5, 6, 8, and 22 h post-treatment, and were analyzed for scopolamine levels using radioreceptor assay. Significantly higher plasma scopolamine levels were found in group 1 at 0.5, 1, 1.5, and 2.5 h, and in group 2 at 1 and 1.5 h post-treatment, compared with group 3. Thereafter, plasma levels did not differ significantly between the groups. In all subjects of group 1 and seven subjects (88%) of group 2, therapeutic levels (>50 pg/ml) were measured during the first 2.5 h, compared with only two subjects (25%) of group 3 (P < 0.05). Heart rate, blood pressure, visual accommodation, performance test results, and subjective complaints of adverse effects did not differ significantly. The combination of transdermal and oral scopolamine (0.3 or 0.6 mg) provides the required plasma levels to prevent seasickness, starting as early as 0.5 h post-treatment, with no significant adverse effects.

An animal model for studying therapeutic drugs against post-traumatic stress disorder.

An animal model for the evolvement of post-traumatic stress disorder (PTSD) was developed by simulating the hormonal consequences of prolonged stress via the continuous administration of corticosterone by subcutaneously implanted sustained-release pellets. Behavioral, morphological, and biochemical effects were recorded and analyzed. This model has shown cognitive deficits as well as hippocampal damage in the rat similar to those found in PTSD patients. The model was also used to test a therapeutic treatment against stress-induced brain damages. Concomitant treatment with the L-type calcium channel blocker, nimodipine, protected young rats from corticosterone-induced morphological brain changes but not cognitive impairments. The proposed animal model may be useful for testing the efficacy of various neuroprotective drugs. Development of an effective drug treatment for use after a traumatic event and through the trauma period might prevent permanent brain damage and the development of PTSD.

Kinetics of brain cholinesterase inhibition following metrifonate administration.

In previous metrifonate (MTF) studies, there has been evidence for a preferential functional effect of the drug in cortical but not in striatal regions. In the present study we investigated the kinetics of brain cholinesterase (ChE) inhibition following an acute administration of MTF (100 mg/kg) in various brain regions of young and old Fischer 344 rats. The main objective was to test the hypothesis that the functional regional selectivity, observed in previous studies, was correlated with the extent of ChE inhibition. Using Karnovsky's method for histochemical staining, the highest staining intensity in control rats was found in the striatum and hippocampus, compared to a low basal activity in the frontal and frontoparietal cortices. In the striatum of drug treated old rats, enzyme inhibition was somewhat greater than that found in young rats. However, in the hippocampus, four to eight hours following MTF administration, the inhibition was greater in young compared to old rats. The differences in the sensitivity of various brain regions towards MTF induced ChE inhibition could not be correlated with the regional variation of MTF functional effects.

Sub-regional hippocampal vulnerability in various animal models leading to cognitive dysfunction.

Various animal models, involving different brain insults, lead to memory deficits, which can be measured using behavioral tests. In numerous studies, using five different experimental models in rats, we have found that cognitive dysfunction is invariably accompanied by hippocampal CA1 and CA3 pyramidal cells degeneration. However, of these two, the most affected area changes from one model to the other. The present manuscript describes and compares the morphological alterations within the hippocampus in the following experimental models: normal aging, hypoxia, prolonged corticosterone administration, brain ischemia and cholinesterase (ChE) inhibition. In all the above, many hippocampal neurons were severely damaged, however, CA3 pyramidal cells were mostly affected in normal aging and following hypobaric hypoxia, whereas CA1 cells were especially affected following corticosterone administration, global ischemia and ChE inhibition. Several mechanisms, which might be involved in the diverse courses of the lesions are being considered: cerebral oxygen and glucose, glutamate neurotoxicity and calcium involvement. It is anticipated that elucidation of the specific role of CA1 and CA3 hippocampal sub-fields in the various experimental models might help in understanding processes such as age-related neuronal degeneration and assist in their prevention.

Nimodipine's protection against corticosterone-induced morphological changes in the hippocampus of young rats.

Sustained high levels of corticosterone (CORT), one of the major stress-induced hormones in the rat, were suggested as generating 'accelerated brain aging' and were shown to induce both specific brain changes in the hippocampus and learning impairments in young and middle-aged Fischer-344 rats. Evidence that altered calcium (Ca) homeostasis may play a major role in brain aging has accumulated over the last decade. Recently, new data established a connection between glucocorticoids and voltage-activated Ca influx in aged hippocampal neurons. In the present study, an attempt was made to block the CORT-induced 'accelerated aging' by the simultaneous administration of the L-type Ca channel blocker nimodipine. CORT or placebo sustained-release (SR) pellets were implanted subcutaneously in 3 months old Fischer male rats. Each group was further sub-divided between nimodipine and placebo SR treatments. Characteristic CORT-induced morphological changes were observed in pyramidal hippocampal cells, such as at the CA1 and CA4 sub-regions (22.2% +/- 7.7 and 28.6% +/- 8.4 of pyknotic cells without clear nuclei, respectively). Concomitant treatment with nimodipine conferred full protection against CORT-induced morphological changes (e.g. 3.2% +/- 0.8 and 2.1% +/- 1.9 of pyknotic cells in CA1 and CA4, n = 7 rats in each group; P < 0.04). The neuroprotective efficacy of nimodipine supports the theory of Ca involvement in CORT related 'accelerated brain aging'.

Effects of metrifonate on radial arm maze acquisition in middle-aged rats.

The efficacy of metrifonate, a well-tolerated cholinesterase (ChE) inhibitor, in attenuating the normal aging- and corticosterone-induced impairments of radial maze performance of rats was compared. Middle-aged Fischer 344 rats were screened for their spatial orientation performance in the Morris water escape task. Good and bad performers were selected: good performers (N= 22) were treated with subcutaneous sustained-release corticosterone pellets, resulting in hippocampal cell damage and impaired spatial orientation in the radial maze; age-induced bad performers (N = 20) were tested without additional pharmacological intervention. Metrifonate (MFT), administered daily during radial maze testing, 30 min before training, at a dose of 15 mg/kg p.o., facilitated the acquisition of the task in age-impaired rats, but not in corticosterone-impaired rats.

Nimodipine counteracts corticosterone-induced habituation impairments.

Corticosterone or placebo sustained-release pellets (4 pellets of 200 mg each, released over 90 days) were implanted subcutaneously in young Fischer-344 rats, fed with either regular food or with food containing 860 ppm of nimodipine. Following 2 weeks of treatment, the habituation of the rats to a new environment was studied. On the first test day, placebo-implanted rats explored the new environment and exhibited a characteristic habituation. On the second test day, 48 hr later, low activity was measured in the already familiar environment. This habituation was absent in corticosterone-implanted rats fed with regular food. However, corticosterone-implanted rats fed with food containing nimodipine behaved during the second test similarly to the placebo-implanted group. The data indicated that the behavioral deficit, induced in Fischer-344 rats by the high corticosterone levels, was reversed by the nimodipine treatment. Thus, nimodipine may be useful in counteracting certain prolonged stress-related cognitive impairments.

Aging, stress, and cognitive function.

Stress was implied as involved in "enhanced aging," and prolonged administration of corticosterone was claimed to lead to central neuronal lesions. This study describes an animal model that simulates the steroid elevation associated with stress by a continuous slow-release administration of corticosterone, in young (3 months old) and middle-aged (12 months old) Fischer 344 rats. Plasma concentrations of corticosterone were stable throughout the day, with no diurnal variation, within the range associated with mild stress. Corticosterone prolonged treatment resulted in morphological changes mainly in the CA1, CA4, and dentate gyrus areas of the hippocampus. Middle-aged rats showed higher vulnerability to the long-term COR treatment than young ones, even when COR treatment was prolonged in young rats from 63 to 90 days. Middle-aged rats were screened before the corticosterone treatment, using the Morris water maze, and divided between cognitively "impaired" and "nonimpaired" subpopulations. Severe cognitive damage during acquisition of the eight-arm radial maze was shown, after the continuous hormonal treatment, in rats initially defined as "nonimpaired" in the Morris water maze. This animal model might be useful for testing the protective effects of drugs against brain changes and cognitive damage, during either pathological or normal aging.

Transdermal physostigmine in the treatment of Alzheimer's disease.

Physostigmine has been reported to improve the memory function of some patients with Alzheimer's Disease (AD). However, the drug has a short half-life and a narrow therapeutic window. To overcome these impediments, we developed a continuous transdermal delivery system and tested it for 2 weeks in 12 AD inpatients, using a single-blind design. No major adverse effects were recorded in any of the patients. Physostigmine plasma concentrations were relatively stable (0.56 +/- 0.10 ng/ml) and correlated well with blood acetylcholinesterase inhibition. Six of the 12 patients reported improved vigilance and concentration, and also had higher scores in all four neuropsychological tests employed (Mini Mental State examination, Short Mental Test [SMT], Wechsler's Memory Scale [WMS], and Buschke's Selective Reminding Test). The performance of two additional patients improved in only two tests (SMT and WMS). Transdermal delivery of physostigmine appears to be safe and may be useful for the treatment of a subset of AD patients.

Cognitive deficits induced in young rats by long-term corticosterone administration.

Corticosterone slow-release pellets, implanted for 9 weeks in young Fischer 344 rats, resulted in continuous high plasma levels of the hormone which are comparable to those of rats under mild stress. One week following termination of the drug treatment, the rats were tested in an eight-arm radial maze. During the initial acquisition stages, corticosterone-treated rats exhibited cognitive impairments in contrast to placebo-treated rats. The deficits were observed in all three parameters which were monitored, the total number of errors, the number of correct entries out of the first eight, and the total time needed to complete the test. This study is the first to report specific behavioral decrements related to the previously observed morphological hippocampal changes induced by long-term corticosterone administration.

Reversal of age-related cognitive impairments by an M1 cholinergic agonist, AF102B.

This study examined the effect of a specific M1 cholinergic agonist, AF102B, on place learning of aged and young rats. Spatial reference memory was tested in the Morris Water Maze task, while spatial working memory was tested on an 8-arm radial maze. Both memory functions were impaired in aged rats compared to young animals. However, the administration of AF102B significantly reduced the age-related cognitive impairments observed in both tasks. This data supports the assertion of the "cholinergic hypothesis," namely that specific enhancement of cholinergic function may reverse geriatric cognitive deficits.

(+-)-cis-2-methyl-spiro(1,3-oxathiolane-5,3') quinuclidine (AF102B): a new M1 agonist attenuates cognitive dysfunctions in AF64A-treated rats.

(+-)-cis-2-Methyl-spiro(1,3-oxathiolane-5,3')quinuclidine (AF102B), a new muscarinic agonist of utmost rigidity, exhibits a high selectivity for M1 muscarinic receptors. In rats having a cholinergic hypofunction induced by the intracerebroventricular administration of ethylcholine aziridinium (AF64A), AF102B reversed cognitive impairments in a step-through passive avoidance task and in an 8-arm radial maze. AF102B reversed cognitive impairments at significantly lower doses than those needed to induce side-effects. In addition, AF102B exhibited low toxicity. The results suggest that AF102B may prove useful for treatments of cholinergic deficiencies and cognitive impairments, like those reported in Alzheimer's disease.

Temporal relationship of the pulsatile fluctuation of luteinizing hormone and progesterone in cattle: a time series cross-correlation analysis.

The temporal relationship between the pulsatile patterns of plasma luteinizing hormone (LH) and progesterone (P4) was studied in mid-luteal (ML) and early-pregnant (EP) dairy cows. Blood samples were collected (via external jugular vein cannulae) at 10-min intervals for 16 h in 5 ML cows (d 10 to 12 of the cycle) and for 10 h in 5 EP cows (d 52 to 56 of gestation). Concentrations of LH and P4 were determined by radioimmunoassays and a time series cross-correlation analysis was utilized to evaluate the temporal relationship between them. A pulsatile pattern was found for both hormones in both groups, and in all animals LH peaks were uniformly followed by P4 peaks. In 80% of the cows in both groups the highest cross-correlation occurred between samples LH(n) and P4 (n + 1) (n = sample number), suggesting that a lag time of about 10 min is necessary for luteal stimulation. Results from both groups demonstrate that P4 is released from the corpus luteum in a pulsatile manner and that its release is at least partially dependent upon the pulsatile pattern of plasma LH. Correlation coefficients between LH and P4 mean level, basal levels, peak frequencies and peak amplitudes obtained in both groups indicate that the conceptus alters the hypothalamic-pituitary-ovarian relationship found in the ML cows, suggesting the existence of another factor(s) acting along with LH to release P4 in EP cows.

Ovarian activity of Holstein and Jersey cows of diverse transmitting abilities for milk.

The relationship between ovarian activity and milk yield was studied in 35 daughters of 24 Holstein sires and 17 daughters of 14 Jersey sires in the same herd. Ovulations and length of estrous cycles were determined by progesterone concentration in postmilking strippings three times per week, by weekly palpation per rectum, and by twice daily estrus detection. Transmitting abilities were for 4% fat-corrected milk of cows and their sires. Yields of 4% fat-corrected milk were estimated for 60, 90, 120, and 305 days in lactation. Postpartum intervals to first ovulation averaged 22 and 20 days for Holsteins and Jerseys, but the interval to first standing estrus was shorter for Jersey than for Holstein. Postpartum intervals to each of the first three ovulations and length of estrous cycles were unrelated to actual yield or transmitting ability for yield of 4% fat-corrected milk in either breed. The percentage of cows observed in standing estrus at each of the first three ovulations increased from 23 to 43%.