Spontaneous persistent activity and inactivity in vivo reveals differential cortico-entorhinal functional connectivity.

Understanding the functional connectivity between brain regions and its emergent dynamics is a central challenge. Here we present a theory-experiment hybrid approach involving iteration between a minimal computational model and in vivo electrophysiological measurements. Our model not only predicted spontaneous persistent activity (SPA) during Up-Down-State oscillations, but also inactivity (SPI), which has never been reported. These were confirmed in vivo in the membrane potential of neurons, especially from layer 3 of the medial and lateral entorhinal cortices. The data was then used to constrain two free parameters, yielding a unique, experimentally determined model for each neuron. Analytic and computational analysis of the model generated a dozen quantitative predictions about network dynamics, which were all confirmed in vivo to high accuracy. Our technique predicted functional connectivity; e. g. the recurrent excitation is stronger in the medial than lateral entorhinal cortex. This too was confirmed with connectomics data. This technique uncovers how differential cortico-entorhinal dialogue generates SPA and SPI, which could form an energetically efficient working-memory substrate and influence the consolidation of memories during sleep. More broadly, our procedure can reveal the functional connectivity of large networks and a theory of their emergent dynamics.

Cortical alterations in a model for absence epilepsy and febrile seizures: in vivo findings in mice carrying a human GABA(A)R gamma2 subunit mutation.

Childhood absence epilepsy (CAE) is one of the most common forms of epilepsy among children. The study of a large Australian family demonstrated that a point mutation in the gene encoding the gamma2 subunit of the GABA(A) receptor (G2R43Q) leads to an autosomal dominantly inherited form of CAE and febrile seizures (FS). In a transgenic mouse model carrying the gamma2 (R43Q) mutation heterozygous animals recapitulate the human phenotype. In-vitro experiments indicated that this point mutation impairs cortical inhibition and thus increases the likelihood of seizures. Here, using whole-cell (WC) and extracellular (EC) recordings as well as voltage-sensitive dye imaging (VSDI), we systematically searched for an in vivo correlate of cortical alterations caused by the G2R43Q mutation, as suggested by the mentioned in vitro results. We measured spontaneous and whisker-evoked activity in the primary somatosensory cortex and ventral posteriomedial nucleus of the thalamus (VPM) before and after intraperitoneal injection of the ictogenic substance pentylenetetrazol (PTZ) in urethane-anesthetized G2R43Q mice and controls in a blinded setting. Compared to wildtype controls in G2R43Q mice after PTZ injection we found 1.) Increased cortical spontaneous activity in layer 2/3 and layer 5/6 pyramidal neurons (increased standard deviation of the mean membrane potential in WC recordings), 2.) Increased variance of stimulus evoked cortical responses in VSDI experiments. 3.) The cortical effects are not due to increased strength or precision of thalamic output. In summary our findings support the hypothesis of a cortical pathology in this mouse model of human genetic absence epilepsy. Further study is needed to characterize underlying molecular mechanisms.

Spontaneous persistent activity in entorhinal cortex modulates cortico-hippocampal interaction in vivo.

Persistent activity is thought to mediate working memory during behavior. Can it also occur during sleep? We found that the membrane potential of medial entorhinal cortex layer III (MECIII) neurons, a gateway between neocortex and hippocampus, showed spontaneous, stochastic persistent activity in vivo in mice during Up-Down state oscillations (UDS). This persistent activity was locked to the neocortical Up states with a short delay, but persisted over several cortical UDS cycles. Lateral entorhinal neurons did not show substantial persistence, and current injections similar to those used in vitro failed to elicit persistence in vivo, implicating network mechanisms. Hippocampal CA1 neurons' spiking activity was reduced during neocortical Up states, but was increased during MECIII persistent states. These results provide, to the best of our knowledge, the first direct evidence for persistent activity in MECIII neurons in vivo and reveal its contribution to cortico-hippocampal interaction that could be involved in working memory and learning of long behavioral sequences during behavior, and memory consolidation during sleep.

Explicit-duration hidden Markov model inference of UP-DOWN states from continuous signals.

Neocortical neurons show UP-DOWN state (UDS) oscillations under a variety of conditions. These UDS have been extensively studied because of the insight they can yield into the functioning of cortical networks, and their proposed role in putative memory formation. A key element in these studies is determining the precise duration and timing of the UDS. These states are typically determined from the membrane potential of one or a small number of cells, which is often not sufficient to reliably estimate the state of an ensemble of neocortical neurons. The local field potential (LFP) provides an attractive method for determining the state of a patch of cortex with high spatio-temporal resolution; however current methods for inferring UDS from LFP signals lack the robustness and flexibility to be applicable when UDS properties may vary substantially within and across experiments. Here we present an explicit-duration hidden Markov model (EDHMM) framework that is sufficiently general to allow statistically principled inference of UDS from different types of signals (membrane potential, LFP, EEG), combinations of signals (e.g., multichannel LFP recordings) and signal features over long recordings where substantial non-stationarities are present. Using cortical LFPs recorded from urethane-anesthetized mice, we demonstrate that the proposed method allows robust inference of UDS. To illustrate the flexibility of the algorithm we show that it performs well on EEG recordings as well. We then validate these results using simultaneous recordings of the LFP and membrane potential (MP) of nearby cortical neurons, showing that our method offers significant improvements over standard methods. These results could be useful for determining functional connectivity of different brain regions, as well as understanding network dynamics.

Mosquito larvicidal properties of aqueous and acetone extracts of Trianthema portulacastrum Linn. (family: Aizoaceae) against vector species of mosquitoes.

Crude aqueous and acetone extracts of leaf of Trianthema portulacastrum was evaluated for its mosquito larvicidal properties against the larvae of four vector species of mosquito under laboratory conditions. These extracts showed good larvicidal properties, 100% mortality in the third instar larvae was observed in the larval bioassay test with Anopheles culicifacies, Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti at a concentration of 1.0, 0.75, 0.75 and 1.0% respectively. The LD50 values estimated for aqueous extract against the larvae of An. culicifacies, An. stephensi, Cx. quinquefasciatus and Ae. aegypti were 0.17, 0.15, 0.18 and 0.17% and LD90 values were 0.57, 0.58, 0.76, and 0.75% respectively. The LD50 values estimated for acetone extract against the larvae of An. culicifacies, An. stephensi, Cx. quinquefasciatus and Ae. aegypti were 0.09, 0.10, 0.09 and 0.12 % and LD90 values were 0.28, 0.25, 0.26 and 0.26% respectively. It is observed that the acetone extract is more effective as compared to aqueous extract. Trianthema portulacastrum can be considered as a probable source of photochemical used for the development of larvicide against disease vectors.

Sensory experience alters specific branches of individual corticocortical axons during development.

Sensory experience can, over the course of days to weeks, produce long-lasting changes in brain function. Recent studies suggest that functional plasticity is mediated by alterations of the strengths of existing synapses or dynamics of dendritic spines. Alterations of cortical axons could also contribute to functional changes, but little is known about the effects of experience at the level of individual corticocortical axons. We reconstructed individual layer (L) 2/3 pyramidal neurons filled in vivo in developing barrel cortex of control and partially sensory-deprived rats. L2 axons had larger field spans than L3 axons but were otherwise equivalently affected by deprivation. Whisker trimming over approximately 2 weeks markedly reduced overall length of axonal branches in L2/3, but individual horizontal axons were as likely to innervate deprived areas as spared ones. The largest effect of deprivation was instead to reduce the length of those axonal branches in L2/3 oriented toward deprived regions. Thus, the location of a branch relative to its originating soma, rather than its own location within any specific cortical column, was the strongest determinant of axonal organization. Individual axons from L2/3 into L5/6 were similarly altered by whisker trimming although to a lesser extent. Thus, sensory experience over relatively short timescales may change the patterning of specific axonal branches within as well as between cortical columns during development.

Differential responses of hippocampal subfields to cortical up-down states.

The connectivity of the hippocampal trisynaptic circuit, formed by the dentate gyrus, the CA3 and the CA1 region, is well characterized anatomically and functionally in vitro. The functional connectivity of this circuit in vivo remains to be understood. Toward this goal, we investigated the influence of the spontaneous, synchronized oscillations in the neocortical local field potential, reflecting up-down states (UDS) of cortical neurons, on the hippocampus. We simultaneously measured the extracellular local field potential in association cortex and the membrane potential of identified hippocampal excitatory neurons in anesthetized mice. Dentate gyrus granule cells showed clear UDS modulation that was phase locked to cortical UDS with a short delay. In contrast, CA3 pyramidal neurons showed mixed UDS modulation, such that some cells were depolarized during the cortical up state and others were hyperpolarized. CA1 pyramidal neurons, located farther downstream, showed consistent UDS modulation, such that when the cortical and dentate gyrus neurons were depolarized, the CA1 pyramidal cells were hyperpolarized. These results demonstrate the differential functional connectivity between neocortex and hippocampal subfields during UDS oscillations.

Phase-locking of hippocampal interneurons' membrane potential to neocortical up-down states.

During quiet wakefulness and sleep, and under anesthesia, the membrane potentials of neocortical pyramidal neurons show synchronous, slow oscillations, so-called up-down states (UDS), that can be detected in the local field potential (LFP). The influence of this synchronized, spontaneous neocortical activity on the hippocampus is largely unknown. We performed the first in vivo whole-cell recordings from hippocampal dorsal CA1 interneurons and found that their membrane potentials were phase-locked to neocortical up-down states with a small delay. These results provide strong evidence for cortico-hippocampal interaction and suggest that neocortical activity drives hippocampal interneurons during UDS.

Entomological indices of Aedes aegypti at some international airports and seaports of southern India--a report.

Entomological surveys were undertaken at some of the international airports/seaports (Bangalore, Calicut, Chennai, Cochin, Thiruvanathapuram and Vishakapatnam) to find out the breeding prevalence of dengue vector mosquito in diverse breeding containers from 1998 to 2004. Three vector indices (House index, Container index and Breateu index) were used to assess the breeding potential at each airport/seaport. International Health Regulations urged national governments to keep all the international airports/seaports and peripheral areas up to 400 meters free from Aedes aegypti mosquitoes. However, surveys revealed high vector indices at all the airports and seaports. Health authorities of airports/seaports need to take cognizance of these facts and develop action plan for appropriate control measures with emphasis on vector surveillance.

Synaptic changes in layer 2/3 underlying map plasticity of developing barrel cortex.

The functional and anatomical rearrangements of cortical sensory maps accompanying changes in experience are not well understood. We examined in vivo and in vitro how the sensory map and underlying synaptic connectivity of the developing rat barrel cortex are altered when the sensory input to the cortex is partially deprived. In the nondeprived cortex, both the sensory responses and synaptic connectivity between columns were strengthened through an increase in the synaptic connection probability between L2/3 pyramids in adjacent columns. This was accompanied by a selective growth of L2/3pyramid axonal arbors between spared columns. In contrast, deprived and nondeprived cortical columns became weakly connected in their L2/3 pyramid connections.

Five-year surveillance of acute flaccid paralysis in Malaysia.

OBJECTIVE: The nation-wide surveillance for acute flaccid paralysis (AFP) was implemented in Malaysia in 1995 and further intensified in 1996 as part of the World Health Organization's (WHO) certification process for polio eradication in the Western Pacific Region. Clinical data on AFP cases during a 5-year surveillance period from 1997 to 2001 were compiled and analysed. RESULTS: Based on 517 cases of AFP reported during this 5-year period, the overall rate of AFP was 1.2 per 100 000 children below 15 years old. The major clinical diagnosis associated with AFP were Guillain-Barre syndrome (30.2%), central nervous system infection (16.2%), transverse myelitis (10.6%) non-polio enterovirus infection (6.2%), and hypokalaemic paralysis (5.2%). This unusual pattern with an excess of CNS infection and non-polio enterovirus infection was attributed to the outbreak of enterovirus 71 infection nation-wide in 1997. According to the WHO virological classification, there was no case of poliomyelitis due to wild poliovirus. Three cases were 'polio compatible', there were no cases of vaccine-associated paralytic polio (VAPP), while 62 cases (12.0%) were merely classified as 'non-polio AFP'. CONCLUSION: Overall, these data suggest the absence of circulation of wild poliovirus in Malaysia from 1997 to 2001. The pattern of AFP in this study is different from other published reports.

Outbreak investigation of scrub Typhus in Himachal Pradesh (India).

Scrub Typhus outbreak investigation was undertaken during September 2003 in the three worst affected districts Shimla, Solan and Sirmaur in Himachal Pradesh (India). A total of 113 cases and 19 deaths (17.27 percent case fatality rate) were reported from the eight districts, which were reporting cases. Cases were prevalent in all age groups and in both the sexes among the persons frequenting forest for occupational work. 35.7 percent of the patients serum samples showed a titer of > 1: 80 against OX 19 and OXK antigen is suggestive of Scrub Typhus. Entomologically Rattus rattus (39.5 per cent) was the most prevalent species followed by Suncus murinus (22.91 percent), Bandicoota bengalensis (29.16 percent) and Bandicoota indica and Mus musculus (4.16 percent each). Vector species Leptotrombidium deliense and Gahrliepia (schoengastilla) sp. were recorded. The chigger index 23.0 was found to be highest in Vill. Baldian and 5.0 in Vill. Bhatakuffer (Shimla) and 1.0 in Vill. Rebon (Solan), which is above the critical limit of 0.69 per rodents. Other mite species, ixodid ticks, fleas and lice have been recorded.

Interaction of sensory responses with spontaneous depolarization in layer 2/3 barrel cortex.

The rodent primary somatosensory cortex is spontaneously active in the form of locally synchronous membrane depolarizations (UP states) separated by quiescent hyperpolarized periods (DOWN states) both under anesthesia and during quiet wakefulness. In vivo whole-cell recordings and tetrode unit recordings were combined with voltage-sensitive dye imaging to analyze the relationship of the activity of individual pyramidal neurons in layer 2/3 to the ensemble spatiotemporal dynamics of the spontaneous depolarizations. These were either brief and localized to an area of a barrel column or occurred as propagating waves dependent on local glutamatergic synaptic transmission in layer 2/3. Spontaneous activity inhibited the sensory responses evoked by whisker deflection, accounting almost entirely for the large trial-to-trial variability of sensory-evoked postsynaptic potentials and action potentials. Subthreshold sensory synaptic responses evoked while a cortical area was spontaneously depolarized were smaller, briefer and spatially more confined. Surprisingly, whisker deflections evoked fewer action potentials during the spontaneous depolarizations despite neurons being closer to threshold. The ongoing spontaneous activity thus regulates the amplitude and the time-dependent spread of the sensory response in layer 2/3 barrel cortex.

Characterization of oxidative stress in various tissues of diabetic and galactose-fed rats.

Rats fed a galactose-rich diet have been used for several years as a model for diabetes to study, particularly in the eye, the effects of excess blood hexoses. This study sought to determine the utility of galactosemia as a model for oxidative stress in extraocular tissues by examining biomarkers of oxidative stress in galactose-fed rats and experimentally-induced diabetic rats. Sprague-Dawley rats were divided into four groups: experimental control; streptozotocin-induced diabetic; insulin-treated diabetic; and galactose-fed. The rats were maintained on these regimens for 30 days, at which point the activities of catalase, glutathione peroxidase, glutathione reductase, and superoxide dismutase, as well as levels of lipid peroxidation and reduced and oxidized glutathione were determined in heart, liver, and kidney. This study indicates that while there are some similarities between galactosemic and diabetic rats in these measured indices of oxidative stress (hepatic catalase activity levels and hepatic and renal levels of oxidized glutathione in both diabetic and galactosemic rats were significantly decreased when compared to normal), overall the galactosemic rat model is not closely parallel to the diabetic rat model in extra-ocular tissues. In addition, several effects of diabetes (increased hepatic glutathione peroxidase activity, increased superoxide dismutase activity in kidney and heart, decreased renal and increased cardiac catalase activity) were not mimicked in galactosemic rats, and glutathione concentration in both liver and heart was affected in opposite ways in diabetic rats and galactose-fed rats. Insulin treatment reversed/prevented the activity changes in renal and cardiac superoxide dismutase, renal and cardiac catalase, and hepatic glutathione peroxidase as well as the hepatic changes in lipid peroxidation and reduced and oxidized glutathione, and the increase in cardiac glutathione. Thus, prudence should be exercised in the use of experimentally galactosemic rats as a model for diabetes until the correspondence of the models has been more fully characterized.

Insecticide susceptibility status of Culex tritaeniorhynchus giles, vector of Japanese encephalitis in Delhi.

Laboratory studies were carried out to ascertain the current susceptibility status of adult and larval stages of the Culex tritaeniorhynchus mosquito, vector of Japanese encephalitis, to various insecticides used under public health programs in India. The present study revealed that exposure of adult mosquitoes to diagnostic concentrations of DDT - 4.0%, malathion - 5.0%, fenitrothion - 1.0%, and propoxur - 0.1% could induce only 50.0, 10. 0, 15.0, and 5.0% mortality, respectively, indicating that the species was resistant to all of these insecticides. The LT50 and LT95 values calculated using diagnostic concentrations of DDT, malathion, fenitrothion, and propoxur were found to be 56.4 and 136, 138 and 272, 185 and 258, and 187 and 249 min, respectively. However, when adult mosquitoes were exposed to the diagnostic concentration of synthetic pyrethroids, viz., deltamethrin - 0.025%, permethrin - 0.25%, and lambdacyhalothrin - 0.1%, 100.0% mortality was observed, indicating that the species was highly susceptible to these adulticides. Larval susceptibility tests carried out using diagnostic dosages of DDT- 0.008, temephos- 0.02, fenthion- 0.008, fenitrothion- 0.125, and malathion- 0.005 mg/l failed to induce any mortality, indicating that larvae were resistant to these larvicides. The LC50 and LC90 values calculated for commonly used larvicides, viz., temephos and fenthion, were 0.1511 and 1.9098, and 0.6151 and 2.395 mg/l, respectively. Increase in tolerance level were estimated at 95.5- and 299.4-fold when these LC90 values were compared with diagnostic dosages of temephos and fenthion, respectively.