Assessing vulnerability to embolism and hydraulic safety margins in reed-like Restionaceae.

The African Restionaceae (Poales), the dominant graminoid layer in the megadiverse Cape Floristic Region of South Africa, are distributed across a wide range of moisture availability, yet currently there is very little known about the underlying hydraulics of this group. We tested two methods for measuring culm vulnerability to embolism, the optical and pneumatic methods, in three species of Cannomois ranging in habitat from semi-riparian (Cannomois virgata) to dryland (Cannomois parviflora and C. congesta). Estimates of culm xylem vulnerability were coupled with measures of turgor loss point (ΨTLP) and minimum field water potential (ΨMD) to assess hydraulic safety margins. The optical and pneumatic methods produced similar estimates of P50, but differed for P12 and P88. All three species were quite vulnerable to embolism, with P50 of -1.9 MPa (C. virgata), -2.3 MPa (C. congesta), and -2.4 MPa (C. parviflora). Estimates of P50, ΨTLP and ΨMD aligned with habitat moisture stress, with highest values found in the semi-riparian C. virgata. Consistent differences in P50, ΨMD and ΨTLP between species resulted in consistent hydraulic safety margins across species of 0.96 ± 0.1 MPa between ΨMD and P50, with onset of embolism occurring 0.43 ± 0.04 MPa after ΨTLP for all three species. Our study demonstrates that restio occupancy of dry environments involves more than the evolution of highly resistant xylem, suggesting that other aspects of water relations are key to understanding trait-environment relationships in this group.

Effects of Strain Rate and Temperature on the Mechanical Properties of Simulated Silica Ionogels.

Ionogels are hybrid materials formed by impregnating the pore space of a solid matrix with a conducting ionic liquid. By combining the properties of both component materials, ionogels can act as self-supporting electrolytes in Li batteries. In this study, molecular dynamics simulations are used to investigate the dependence of mechanical properties of silica ionogels on solid fraction, temperature, and pore width. Comparisons are made with corresponding aerogels. We find that the solid matrix fraction increases the moduli and strength of the ionogel. This varies nonlinearly with temperature and strain rate, according to the contribution of the viscous ionic liquid to resisting deformation. Owing to the temperature and strain sensitivity of the ionic liquid viscosity, the mechanical properties approach a linear mixing law at high temperature and low strain rates. The median pore width of the solid matrix plays a complex role, with its influence varying qualitatively with deformation mode. Narrower pores increase the relevant elastic modulus under shear and uniaxial compression but reduce the modulus obtained under uniaxial tension. Conversely, shear and tensile strength are increased by narrowing the pore width. All of these pore size effects become more pronounced as the silica fraction increases. Pore size effects, similar to the effects of temperature and strain rate, are linked to the ease of fluid redistribution within the pore space during deformation-induced changes in the geometry of the pores.

Computational Study of the Structure and Transport in Pyrrolidinium-Li-TFSI-Silica Ionogels.

Ionogels (IGs) are a unique class of composite materials with attributes that make them promising materials for applications in electrochemical energy storage. Due to the solid porous matrix that confines the ionic liquid (IL) in the IG, they can be used as self-supporting electrolytes. Furthermore, interactions of the IL with the porous matrix can have beneficial effects on transport, such as lowering the freezing/glass transition temperature of the conducting IL. In this work, we employ molecular dynamics simulations to investigate the influence of the porous morphology and solid volume fraction on ionic conductivity and Li+ diffusivity using a representative 0.5 M Li-bis(trifluoromethane)sulfonimide (TFSI)-pyrrolidinium (Pyr1.3) IL confined in a nanoporous silica matrix. The effect of the morphology of the confining matrix is compared using the pure IL as a baseline. We find that the tracer and collective Li+ diffusion and ionic conductivity of all the model IGs have significantly lower temperature dependence than the corresponding pure IL. In general, low-silica IGs with wide pores displayed the best transport properties at high temperatures, but the trends with the morphology for the nested set of transport coefficients we examined changed as the collective behavior of the Li+ ions and the molecular IL components were considered. Remarkably, some of the model IGs displayed better transport properties on a volume of fluid basis at low temperatures than the constituent IL. These trends were tied to structural changes revealed by the radial distribution functions of the IL components and the silica surface, including a decreasing Li+ adsorption peak of the surface silica indicating a change in the relative contributions of bulk-like and surface-like transport in the confined IL.

A MALDI sample preparation method suitable for insoluble polymers

Polyamides are insoluble or poorly soluble in common organic solvents, which makes normal sample preparation for matrix-assisted laser desorption/ionization (MALDI) mass spectrometry very difficult. An new analytical protocol for MALDI analysis of polyamides or other insoluble samples is described. It consists of pressing a pellet from a solid mixture of the polymer and a matrix, both in the form of finely ground powder. This sample preparation is compared with the common dried droplet sample preparation method and found to perform much better, both in terms of robustness against variation of experimental parameters and high-mass capability.

Bilateral cerebellar lesions disrupt conditioned eyelid responses in unrestrained rats.

Electromyographic eyelid responses in unrestrained rats were classically conditioned in a Pavlovian delay paradigm by using a tone conditioned stimulus (CS) and periorbital shock unconditioned stimulus (US). After eyelid conditioning was complete, bilateral electrolytic lesions were made in the dentate-interpositus region of the cerebellar nuclei. Initial eyelid conditioning was reliable and very similar to that previously observed in the rabbit, although the asymptotic eyelid responses contained a short-latency startle response in addition to the usual conditioned and unconditioned responses (CR and UR). Substantial decrements in CRs were observed in 13 of the 14 rats with accurately placed lesions. In contrast, startle responses and URs were unaffected. The results replicate the effects of cerebellar lesions on eyelid CRs in the rabbit and suggest that the anatomical basis of eyelid conditioning in both species is similar.

Long-term potentiation facilitates behavioral responding to single-pulse stimulation of the perforant path.

Three experiments were conducted to determine whether long-term potentiation (LTP) could enhance the stimulus properties of electrical brain stimulation. First, a paradigm was developed in which single-pulse stimulation of the perforant path (PP) could acquire control over operant responses. Evoked potentials were recorded from the dentate gyrus (DG) on every trial in order to measure the postsynaptic consequences of the stimulus and to monitor synaptic efficacy in the PP-DG synapses. The second experiment confirmed the relation between the amount of evoked activity and acquisition rate and also showed that transecting the PP impaired performance. In the third experiment, high-frequency stimulation of the PP produced LTP and accelerated subsequent acquisition of behavioral responding to PP stimulation. These results document a clear link between increases in synaptic efficacy and changes in behavior and thereby demonstrate the ability of LTP to serve as at least one component of the neural bases of learning and memory.

Eyeblink conditioning in the developing rat.

Long-Evans rat pups, 17-18 or 24 days of age, were trained with an eyeblink conditioning (EBC) procedure that was used previously with adult rats (Skelton, 1988). Pups received 3 sessions of delay conditioning in a single day at about 4-hr intervals (100 trials/session). Trials involved pairings of an auditory conditioned stimulus (2.8-kHz, 82-dB tone) and a periocular-shock unconditioned stimulus (US; 100 ms, 2 mA), which were presented 280 ms apart. EBC was observed at both ages, but older pups learned much more rapidly. Subsequent experiments established that this effect is associative (Experiment 2), that age differences in EBC cannot be attributed to differences in ability to respond or in sensitivity to the US (Experiment 3), and that EBC rate can be modulated by motivational state (Experiment 4). This preparation may help elucidate the relation between neural development and the ontogeny of learning.

Pharmacological dissociation between the spatial learning deficits produced by morphine and diazepam.

This study sought to determine whether the place learning deficits produced by diazepam are a secondary result of opioid release. Rats pretreated with diazepam (3 mg/kg) or morphine (15 mg/kg) were trained in the Morris water maze. Diazepam impaired place learning-slowing acquisition and preventing the formation of a quadrant preference. Morphine also slowed acquisition, but did not prevent place learning, and impaired escape to a visible platform. Flumazenil blocked the deficits produced by diazepam, but not morphine. Naloxone (2 mg/kg) blocked the deficits produced by morphine, but not diazepam. A high dose of naloxone (10 mg/kg) slowed acquisition, and exacerbated the deficit produced by diazepam. These results demonstrate that diazepam interferes with mnemonic processes through endogenous benzodiazepine receptors, independently of opioidergic systems. Further, they suggest that morphine interferes with motivational processes through opioidergic systems, independently of endogenous benzodiazepine systems.

Like diazepam, CL 218,872, a selective ligand for the benzodiazepine omega 1 receptor subtype, impairs place learning in the Morris water maze.

The sedative, anxiolytic, and amnesic effects of diazepam were compared to those of CL 218,872, a triazolopyridazine that has a preferential affinity for the benzodiazepine omega 1 receptor subtype. Spontaneous locomotion was assessed using a running wheel, anxiety was assessed using an open-field divided into central and peripheral areas (thigmotaxis), and amnesia was assessed using the Morris water maze. It was found that CL 218,872, like diazepam, depressed spontaneous locomotion, reduced anxiety, and impaired place learning in a dose-dependent manner. Flumazenil, a benzodiazepine receptor antagonist with a similar affinity for both omega 1 and omega 2 subtypes, reversed all of the effects of diazepam and antagonized the anxiolytic and amnesic effects, and some but not all of the sedative effects of CL 218,872. These results suggest that the selective activation of the omega 1 receptor subtype by CL 218,872 is sufficient to produce sedation, anxiolysis, and amnesia in a manner similar to that produced by the coactivation of both the omega 1 and omega 2 receptor subtypes with diazepam.

Modelling recovery of cognitive function after traumatic brain injury: spatial navigation in the Morris water maze after complete or partial transections of the perforant path in rats.

The Morris water maze (MWM) has been used to assess cognitive function in rats after a variety of lesions designed to model brain damage and to assess the effects of drugs, growth factors, and neural transplants on post-operative deficits. The present study examined recovery of spatial navigation in the MWM over time in order to model the spontaneous recovery of cognitive function seen in humans. Diffuse axonal injury, a neuropathology commonly associated with traumatic brain injury (TBI), was modelled by transecting the perforant path (PP) bilaterally, either caudal to the hippocampus or dorsal to it at the decussation of the dorsal hippocampal commissure. Both groups with PP cuts showed substantial deficits initially, but spatial performance recovered with time and training. Recovery of platform finding was nearly complete within 14 days of testing, but recovery of platform searching did not occur for 2 or 3 more weeks. When the platform was moved to a new location, a continuing deficit in learning rate was revealed. When the platform was moved to a new position every day, this deficit was even more evident. These results illustrate the multi-faceted nature of recovery after brain injury and provide a new model for assessing the effects of manipulations designed to modulate recovery.

Effects of intracranial infusions of chlordiazepoxide on spatial learning in the Morris water maze. II. Neuropharmacological specificity.

In the preceding paper it was found that infusions of chlordiazepoxide (CDP) into the medial septal region, but not several other regions possessing a high density of benzodiazepine receptors, impaired spatial learning, but not cue learning or swim speed, in the Morris water maze. The present investigation sought to further characterize the neuropharmacological profile of this effect. Initially, it was reconfirmed that systemically administered CDP impaired spatial learning, but not cue learning or swim speed, in the water maze. Additionally, it was found that systemically administered scopolamine, a muscarinic antagonist, impaired both spatial and cue learning, but not swim speed, confirming the detrimental effects of cholinergic hypofunction on maze learning. In new rats, a dose-response assessment revealed that 60 and 30 nmol, but not 10 nmol, CDP infused into the medial septum impaired spatial learning, but not cue learning or swim speed. On the following day, rats from each dose group, now undrugged, acquired a reversed platform location at control levels, suggesting that the previously observed impairment was not due to a neurotoxic effect. Additionally, it was found that systemically administered flumazenil (10 mg/kg) blocked the spatial learning deficit produced by the 60 nmol dose of CDP infused into the medial septum. However, intraseptal infusions of flumazenil (10, 20, or 30 nmol) failed to attenuate the spatial learning deficit produced by systemically administered CDP. Finally, systemically administered tetrahydroaminoacridine (1 or 3 mg/kg), an acetylcholinesterase inhibitor, failed to attenuate the spatial learning deficit produced by intraseptal CDP (60 nmol). Together these results implicate benzodiazepine receptors in the medial septum in the amnesic actions of CDP but suggest that additional sites also mediate this action. The present results fail to support the idea that the spatial learning deficit produced by intraseptal infusions of CDP is due to a suppression of septo-hippocampal cholinergic activity and it is proposed that CDP impairs spatial learning by exacerbating hippocampal inhibition by inhibiting septo-hippocampal GABAergic projection neurons.

Recovery of spatial performance in the Morris water maze following bilateral transection of the fimbria/fornix in rats.

The present study investigated whether spatial performance in the Morris water maze (MWM) recovers after bilateral transection of the fimbria/fornix (FF) in rats, whether such recovery results from restored or residual spatial cognitive capacity, and what contribution, if any, pre-operative training makes to such recovery. Following surgery, rats were administered extensive training to a constant submerged platform location with frequent probe tests to assess performance strategies. Following the attainment of asymptotic performance levels, rats were tested for acquisition of a second platform location. FF lesions were found to produce a severe impairment both in pre-operatively trained rats (a retention or retrieval deficit) and in naive rats (an acquisition deficit) as shown by the use of indirect routes to the platform on submerged platform trials and an absence of localized searching in the platform's area on probe trials. However, with further training, performance recovered in both groups, such that they eventually used direct escape routes to the submerged platform and showed highly localized searching in its area on probe trials. When tested for acquisition of a second platform location, a substantial deficit reappeared, but was again overcome with additional training. Pre-operative training was found to attenuate the initial post-operative deficit and speed recovery of performance but did not affect asymptotic performance levels nor acquisition of the second platform location. These data show that, though spatial cognition as assessed in the MWM is impaired by FF lesions, spatial performance eventually recovers. Moreover, pre-operative training, though of some initial post-operative benefit, is not essential for this recovery. The deficit shown in acquisition of the second platform location argues against recovery of spatial cognition and suggests that the basis of recovered performance is residual spatial cognitive capacity. Several limitations of this residual capacity are apparent: (i) rate of acquisition of spatial information is reduced; (ii) utilization of spatial information stored pre-operatively is restricted; and (iii) translation of spatial information into navigational behaviour is less efficient. The neural bases of this residual system are speculated to include spared intra-hippocampal storage mechanisms and/or mechanisms involved in extra-hippocampal long-term memory consolidation while the neural bases of the FF's contribution to spatial information storage in the intact brain are speculated to involve theta synchronization of hippocampal activity and the induction and expression of hippocampal long-term potentiation.

Dorsal noradrenergic bundle lesions fail to disrupt self-stimulation from the region of locus coeruleus.

Intracranial self-stimulation (ICSS) from the region of the locus coeruleus (LC) was not attenuated by ipsilateral dorsal noradrenergic bundle (DNB) lesions. Bilateral DNB lesions also failed to affect LC ICSS, whereas the same lesions, in the same animals, resulted in a marked enhancement of lateral hypothalamic (LHA) ICSS responding. Behavioral, neuroanatomical and fluorescent histochemical data confirmed destruction of the DNB, thus suggesting that the dorsal noradrenergic system is not necessary for ICSS from the region of the LC.

Kindling with stimulation of the dentate gyrus. I. Characterization of electrographic and behavioral events.

Once daily for 60 days, hooded rats received unilateral high-frequency stimulation in the hilus of the dentate gyrus (DG), at an intensity sufficient to evoke epileptiform afterdischarge (AD). Although most rats eventually developed generalized stage-5 seizures (Generalized group), some did not progress beyond partial stage-1 or stage-2 seizures (Partial group). Hilar kindling also displayed several other characteristics that distinguished it from typical limbic kindling, including low rate of development, marked instability of the seizures, and little or no growth in duration of AD.

Kindling with stimulation of the dentate gyrus. II. Effects on evoked field potentials.

Once daily for 60 days, male hooded rats received unilateral high-frequency stimulation in the hilus of the dentate gyrus (DG), at an intensity sufficient to evoke afterdischarge (AD). Every 2nd day, evoked potentials were recorded from the hilus following stimulation of the PP with single 0.1 ms pulses at 6 current intensities. Changes in synaptic excitability of the dentate granule cells were monitored by measuring the amplitudes of the population spikes; changes in the strength of excitatory synaptic transmission were monitored by measuring the slopes of the excitatory postsynaptic potentials (EPSPs). Control rats, which were not given kindling stimulation, were tested for changes in synaptic transmission and excitability in the same way, at comparable times. In general, hilar stimulation resulted in a large decrease in population spike amplitudes to below baseline and control levels, accompanied by a paradoxical potentiation of EPSPs. Population spike amplitudes decreased more in rats that developed generalized stage-5 seizures (Generalized group) than in rats that did not progress beyond partial seizures despite 60 days of stimulation (Partial group). Conversely, EPSP slopes increased more in the Partial group than in the Generalized group. These results suggest that kindling stimulation may potentiate responsiveness of the directly activated dentate granule cells to inputs from the PP, but at the same time suppress the output of the granule cells resulting from this input. Furthermore, the results indicate that kindling is more closely allied to the suppression of output than to the potentiation of responsiveness to input.

Differential effects of benzodiazepine receptor agonists on hippocampal long-term potentiation and spatial learning in the Morris water maze.

The amnesic effect of benzodiazepine drugs has been well documented, though the mechanisms mediating this effect are unknown. Long-term potentiation (LTP) has been proposed as a mechanism by which information is stored in the mammalian central nervous system. This experiment sought to determine if benzodiazepines impair mnemonic processes by blocking LTP. Rats implanted with a stimulating electrode in the perforant path and a recording electrode in the dentate gyrus were given high-frequency stimulation after the administration of either chlordiazepoxide (5 mg/kg), diazepam (5 mg/kg) or CL 218,872 (10 mg/kg). None of these drugs completely blocked the induction of LTP as measured by changes in the magnitude of the population spike amplitude, though CL 218,872 significantly suppressed potentiation over the duration of recording (24 h). Moreover, the potentiation observed in diazepam-treated rats returned to baseline after 24 h. Two weeks after the last recording, the same implanted rats were given their previous drug and dose and then tested for spatial learning ability in the Morris water maze. Each drug resulted in a severe impairment of spatial learning, but had no effect on cue learning. Two days later, in the absence of drugs, the same rats readily acquired a reversed platform location. Together these results suggest that CL 218,872 may impair spatial learning by suppressing LTP in the perforant path but that chlordiazepoxide and diazepam can impair spatial learning in the absence of LTP suppression in this pathway.

Reference ranges for cardiac dimensions and blood flow velocity in preterm infants.

AIM: To establish reference ranges for cardiac dimensions and Doppler measurements in preterm infants. METHODS: 79 infants of less than 34 weeks' gestation were examined by echocardiography on days 0, 7, and 28 after birth, to produce a set of reference ranges and to examine changes in these indices over the first month of life. The following dimensions were measured: interventricular septum, left ventricular posterior wall, left interventricular diameter at end systole and diastole, left atrium, and aortic root; Doppler measurements were made of maximum blood flow velocity (Vmax) through the pulmonary, aortic, mitral, and tricuspid valves. RESULTS: Reference ranges are given. Cardiac dimensions correlated well with gestation and birth weight but Vmax did not. There was a significant increase in measurements over time. The "normal" preterm infant also appeared to often have asymmetrical septal hypertrophy. Antenatal dexamethasone administration did not appear to affect the measurements. CONCLUSIONS: There is a close correlation with both gestation and birth weight for all physical measurements. Echocardiograms in preterm babies clearly differ from those in older children and adults.

Effect of aging on the spatial learning deficit produced by diazepam in rats.

1. After pretraining in an undrugged state, young (6 months) and aged (18-24 months) rats were trained on a spatial learning-set task after receiving one of four doses of diazepam (1, 2, 3 or 5 mg/kg) or the drug vehicle. The effects of 5 mg/kg of diazepam were also assessed on the spatial learning-set task one full hour after injection (delay condition) as well as on the visible platform task. 2. During pretraining (undrugged), aged rats demonstrated a transient impairment on the visible platform task but subsequently did not differ significantly from young rats on the submerged platform task. On the spatial learning-set task, aged rats performed as well as young rats under control conditions and diazepam produced a comparable dose-dependent impairment of spatial learning. However, when the 1 hr delay was interposed between diazepam administration and maze testing, only aged rats exhibited a spatial learning impairment. Diazepam did not impair performance on the visible platform task in either young or aged rats. 3. These results indicate that although the amnesic effect of diazepam is not initially greater in aged rats, it persists for longer periods.

"Click test": rapid diagnosis of the respiratory distress syndrome.

Appropriate and early treatment with exogenous surfactant has clinical and economic benefits for neonates with pulmonary surfactant deficiency. In order to rapidly and reliably identify such neonates, we have evaluated the shake and click tests, biophysical tests of surfactant function, using 0.2 mL samples of tracheal (TA) and gastric aspirates (GA). Samples from 181 neonates with a gestational age range of 24-40 weeks were shaken with 95% ethanol. If bubbles formed (positive shake test) they were examined in air-free water under a microscope. In a positive shake or click test, the bubbles rhythmically increase and then decrease in size, denoting the presence of active surfactant. The probability of the tests to predict clinical surfactant deficiency was analyzed. The latter was defined as respiratory distress syndrome or transient tachypnea of the newborn diagnosed by chest radiography and clinical criteria. The click test on TA from preterm infants was most accurate, with a 100% positive predictive value and specificity, and a 93% and 94% negative predictive value and sensitivity respectively. These values for GA were 73%, 84%, 97%, and 95%, respectively. The test is quick, simple, inexpensive, reproducible, and unaffected by contamination with blood. The accuracy of this test on TA in diagnosing surfactant deficiency in neonates would permit early and optimal treatment with exogenous surfactant. When performed on GA, the test could aid decisions regarding transfer of neonates to tertiary level care.

HIV in the U.K.: problems of prevalence, sociological response and health education.

It is argued that the prevalence of AIDS is substantially under-represented in existing national AIDS data. Thus although official statistics demonstrate significant recent shifts in transmission routes for HIV, health educators are faced with a problem because it is difficult to develop preventive strategies against a syndrome whose prevalence can only be estimated. Problems of the true extent of the prevalence of HIV are compounded when there is a lack of knowledge about the specifics of heterosexual behaviour. It is unwise to assume that the protective strategies developed by gay men in the face of HIV are routinely available for adoption by heterosexuals, who are characterised by social divisions of age, gender and relative amounts of social power. These concerns represent a problem for health educators. To date, sociological work may not have made the most effective contribution in its support of intervention strategies against HIV/AIDS. Examination of the empirical literature on lay concepts of health and illness reveals a pessimistic stance on the part of some researchers about the ability of individuals to modify behaviour. More positive readings of their own data are possible. The traditional concerns to emphasise the socio-economic determinants of health and behaviour, now also shared by some health educators, should not obscure a concern for the fate of individuals. The most effective contribution that health promotion may be able to make to the control of HIV in the heterosexual population is to assist in the development of strategies of empowerment and 'horizontal intervention'.