Chloroplast capture and range extension after hybridization in taro (Colocasia esculenta).

Complete chloroplast genomes of 17 samples from six species of Colocasia (Araceae) were sequenced, assembled, and aligned together with two previously reported complete genome sequences from taro (Colocasia esculenta). Analysis provides a well-supported phylogenetic tree for taro and closely-related wild Colocasia species in Southeast Asia. Two chloroplast lineages (CI and CII) form a well-defined haplotype group and are found in cultivated taros known as var. esculenta (dasheen, CI), var. antiquorum (eddoe, CII), and in a widespread, commensal wild form known as var. aquatilis (CI). A third lineage (CIII) is also found in wild taros known as var. aquatilis and in the wild species C. lihengiae, C. formosana, and C. spongifolia. We suggest three different scenarios to explain the grouping of CIII wild taros (C. esculenta) with other wild Colocasia species. Chloroplast lineages CI and CIII in C. esculenta and an unknown parent species may be involved in an as yet undated history of hybridization, chloroplast capture, and range extension. Substantial taxonomic revision may be needed for C. esculenta after further studies of morphological and genetic diversity within the crop, in wild populations, and in closely related wild species. The results also point to the Bengal delta as a region of key interest for future research on the origins of tropical wetland taros.

Unambiguous representation of overlapping serial events in the rat hippocampal formation.

The hippocampal formation is suggested to be crucial in unambiguous representation of overlapping temporal sequences in episodic memory. We hypothesized that, if this was true, the hippocampal formation neurons would differentially respond to the same elements even in different temporal sequences. The present study was designed to investigate hippocampal formation CA1 neuronal activity of rats during performance of a conditional delayed stimulus-response association task in which three stimuli were conditionally and serially presented with a delay. In the task, the pairs of the second and third stimuli were overlapped across the trials, but separated by the preceding first stimuli. Conditioned tones coming from one of three possible directions were followed, after a short delay, by one of three pairs of reinforcement series. The pairs consisted of air puff (aversive sensory stimuli) and tube protrusion (which allowed licking sucrose behavior) in the following combinations: air puff-tube protrusion, tube protrusion-tube protrusion and tube protrusion-air puff. The pairs were interposed by a 2 s delay. The three conditioned tone directions were associated with these three pairs in a one-to-one correspondence, and its association was conditional to three possible conditioned tone frequencies (300, 530, and 1,200 Hz). The responses of 107 neurons to the air puff and tube protrusion were analyzed by two-way ANOVA (task condition x reinforcement situation). Of 42 air puff-responsive and 64 tube protrusion-responsive neurons, 36 and 53 displayed significant main effects and/or significant interaction, respectively. Furthermore, neural responses during the delay periods were dependent on the task conditions. The results indicated that the majority of the hippocampal formation neurons showed task condition- and/or reinforcement situation-dependent responses, suggesting a crucial role of the hippocampal formation in representation of overlapping serial events in episodic memory.

Generators of somatosensory evoked potentials investigated by dipole tracing in the monkey.

Generators of somatosensory evoked potentials, elicited by electrical stimulation of the median nerve in anaesthetized monkeys (Macaca fuscata), were investigated by submitting a three-dimensional reconstructed brain model to dipole tracing, which can equate surface potential distributions to an approximate corresponding equivalent dipole. The following components of the somatosensory evoked potentials were simultaneously recorded from 21-27 epidural electrodes: P7 (the letter indicates positive or negative polarity; the number indicates the approximate latency of the peak in ms) was recorded widely from various locations on both the left and right hemispheres, P10 was recorded near the anterior side of the central sulcus contralateral to the stimulation side, N10 was recorded near the posterior side of the contralateral central sulcus, P12 was recorded on both sides of the contralateral central sulcus, and P18 was recorded posterior to the contralateral central sulcus. Current source generators (dipoles) of each component of somatosensory evoked potentials were localized by dipole tracing: a dipole for P7 was located in the thalamus contralateral to the stimulation side; a dipole for P10 and N10 in the posterior wall of the contralateral central sulcus (area 3b); a dipole for P12 in the contralateral post central gyrus (areas 1 and 2); and a dipole for P18 in the anterior wall of the contralateral intraparietal sulcus (area 5). The locations and latencies of dipoles that generated cortical components of somatosensory evoked potentials, estimated by dipole tracing, were confirmed by direct cortical surface recording from a 16-25 electrode array placed directly on the cortical surface; and multiple unit recording from the anterior and posterior parietal cortices. After excision of area 5, P18 and N18 were abolished, whereas P10, N10, and P12 were not affected. The results suggest that dipoles for somatosensory evoked potentials progressed from the thalamus to area 5 via the primary somatosensory area. This progress is consistent with the hierarchical sequence of somatosensory information processing.

Differential activation in the medial temporal lobe during a sound-sequence discrimination task across age in human subjects.

To elucidate the brain mechanisms to encode sequential events, event-related potentials (ERPs) were recorded during a sound-sequence discrimination task using young and middle-aged adult subjects. In the task, a series of six or 12 kinds of natural sounds were sequentially presented; 70-80% of the stimuli were presented in a fixed order (Non-target), but the remaining stimuli, in a random order (Target). The subjects were instructed to detect the Targets and press a button at the end of each Target. In a control task, the same sounds were randomly presented (Control), and they were instructed to press the button at the end of each sound. Behavioral results indicated that the young subjects learned the task faster than did the middle-aged subjects. Positive ERP waves were evoked by Targets and Non-targets in the parieto-occipital area around 300-700 ms after stimulus onset. The mean amplitudes during this period in the young subjects were larger in Target than Control conditions, and those in Target condition were larger in the young than middle-aged subjects. Furthermore, the mean amplitudes in the Target condition were significantly correlated with behavioral performance. Equivalent dipoles for the ERPs evoked by Targets were estimated in the medial temporal lobe including the hippocampal formation and parahippocampal gyrus. The results suggest that the ERPs around 300-700 ms latency are involved in sound-sequence information processing. Furthermore, decrease in amplitudes of this positivity in the middle-aged subjects suggests that age-related memory decline is associated with deficits in encoding and retrieval of unfamiliar sequence.

Comparison of technetium-99m-MIBI, technetium-99m-tetrofosmin, ultrasound and MRI for localization of abnormal parathyroid glands.

UNLABELLED: Abnormal parathyroid tissue can be identified by radionuclide imaging with either 99mTc-MIBI or 99mTc-tetrofosmin. This study compared the relative sensitivity of these two agents to localize parathyroid hyperplasia and adenoma. METHODS: Twenty patients with primary (n = 9) or secondary (n = 11) hyperparathyroidism were studied with 99mTc-MIBI and 99mTc-tetrofosmin parathyroid imaging, ultrasonography and MRI. Radionuclide images of the neck were acquired 10 min and 2-3 hr after radiopharmaceutical injection. The images were visually evaluated for abnormal focal areas of increased tracer localization in the neck and mediastinum. A parathyroid gland/normal thyroid tissue activity ratio (referred to as the P/T uptake ratio) was calculated for each positive scan. RESULTS: Of the 46 parathyroid glands surgically explored, the overall sensitivity and specificity of MIBI imaging were 83% and 83% (38/46); tetrofosmin imaging 87% and 83% (40/46); ultrasonography 78% and 40% (36/46); and MRI 80% and 60% (37/46), respectively. Both radiopharmaceuticals performed well in the nine patients found to have adenoma. The sensitivity and specificity of MIBI imaging were 100% and 100% (9/9); tetrofosmin imaging 100% and 100% (9/9); ultrasonography 78% and 67% (7/9); and MRI 100% and 100% (9/9), respectively. In the 37 glands with hyperplasia, MIBI imaging had a sensitivity of 78% and specificity of 75%; tetrofosmin imaging 84% and 75%; ultrasonography 78% and 43%; and MRI 73% and 60%, respectively. CONCLUSION: All imaging techniques localized abnormal parathyroid glands. The radiotracers have equal sensitivity for the localization of abnormal parathyroid glands. The sensitivity of these tracers was high as compared to ultrasonography or MRI.

Lesions of the medial shell of the nucleus accumbens impair rats in finding larger rewards, but spare reward-seeking behavior.

The goal of this study was to help better understand the importance of the nucleus accumbens (Nacc) in the processing of position and reward value information for goal-directed orientation behaviors. Sixteen male Long-Evans rats, under partial water deprivation, were trained in a plus-maze to find water rewards in the respective arms which were lit in pseudo-random sequence (training trials). Each day one reward arm was selected to deliver six drops of water (at 1 s intervals) the others provided only one drop per visit. After 32 visits, probe trials were intermittently presented among training trials. Here, all four arms were lit and offered the previously assigned reward. The rats rapidly learned to go to the highly rewarded arm. Six trained rats were given bilateral electrolytic lesions in the Nacc shell, two others had unilateral lesions and eight had sham operations (with approved protocols). Field potentials evoked by fornix stimulation were recorded in lesion electrodes to guide placements. Only the lesioned rats showed significant impairments (P<0.05) in selecting the greater reward on probe trials. However on training trials, lesioned (and sham-operated) rats made only rare errors. While the motivation to drink and the capacity for cue-guided goal-directed orientation behavior was spared, lesioned rats were impaired in learning the location of the larger reward. The accumbens lesions apparently impaired integration of position and reward value information, consistent with anatomical and electrophysiological data showing the convergence of hippocampal, amygdalar, ventral tegmental area (VTA) and prefrontal cortical inputs there.

Contribution of chorda tympani and glossopharyngeal nerves to taste preferences of rat for amino acids and NaCl.

To learn how the gustatory nerves convey information about the nutritionally dependent taste preference, intake of amino acid solutions and saline in rats with bilateral chorda tympani (CTX) and/or glossopharyngeal neurotomy (GPX) was determined during the feeding of a control diet (C) and a L-lysine (Lys) deficient diet (LD). Intact rats preferred L-arginine (Arg) more in C and Lys more in LD. The CTX group did not select nor ingest Lys in LD, and its intake of Arg was also low in C. The GPX group did not substantially alter its preference under both diets, while it did show an increase in total liquid intake. The preference changes in the CTX + GPX group appeared as combined effects of the CTX and the GPX groups. In an additional study, the preference for Lys shifted to higher concentrations and the total consumption of Lys increased in LD. The present data suggest that the chorda tympani nerves possibly function as discriminators of the nutritional information by altering the taste preference, and that the glossopharyngeal nerves may convey other functional taste information, such as aversive tastes, and sensory aspects of osmotic regulation. In addition, it is revealed that the animals have ability to search for a nutrient deficient in their body, and to ingest it to a level that at least nullifies the deficiency.

Effects of D2 dopamine receptor agonist and antagonist on brain activity in the rat assessed by functional magnetic resonance imaging.

The effects of D2 dopamine receptor agonist, bromocriptine (BROMO), and antagonist, haloperidol (HPD), on brain activity were investigated in rats by functional magnetic resonance imaging. T2*-weighted signal intensity was increased in the hypothalamus at 120 min after acute administration of BROMO, and in the ventral posterior and dorsomedial nuclei of the thalamus from 30 to 120 min. In contrast, the signal intensity was decreased in the caudate-putamen at 30 min after acute administration of HPD, in the hypothalamus from 30 to 60 min, and in the perirhinal cortex at 30 min. After chronic (2 weeks) HPD treatment, acute administration of HPD decreased signal intensity in the caudate-putamen at 60 min, in the hypothalamus at 30 min, the perirhinal cortex from 2 to 120 min, the dorsomedial and ventral posterior nuclei of the thalamus from 2 to 120 min, and the medial nucleus of the amygdala from 60 to 120 min. These results suggest that (1) the D2 receptor agonist increased the activity of the thalamic nuclei and the hypothalamus, while the D2 receptor antagonist suppressed brain activity in the regions where D2 receptors were present, (2) the suppression of brain activity in the thalamic nuclei and the perirhinal cortex by acute HPD administration was enhanced by chronic HPD treatment, and (3) the effects of antipsychotic drugs on the thalamus, amygdala, and perirhinal cortex may be related to their therapeutic efficacy, since clinical improvement in schizophrenic patients appears several days after the start of HPD treatment.

Neuronal responses in monkey anterior putamen during operant bar-press behavior.

Single neuron activity was recorded in the monkey anterior putamen to compare visuomotor-related responses during operant bar-press behavior with visual discrimination of objects. Of 615 neurons recorded, 9.8% (60/615) responded to the presentation of food during forced delay of access to the bar. Of these 60 neurons, 38 were also tested with nonfood, and 19 of these responded to the nonfood objects regardless of the following movement. The amplitude of the visual-related responses of some differential neurons was graded for different objects to reflect the relative degree of preference for the food presented. However, these responses disappeared in reaction time tasks in which the bar could be accessed for pressing immediately upon presentation of an object. The visual response latency of differential neurons ranged from 50 to 700 ms (mean +/- SD, 386 +/- 211 ms), which was longer than that of the nondifferential responses (207 +/- 204 ms). These results suggest that anterior putamen neurons might participate in estimation of visual information that could be related to forecasting movement.

Ischemic neuronal damage specific to monkey hippocampus: histological investigation.

We previously reported lesions confined specifically to the hippocampus when produced by occluding eight vessels (the bilateral vertebral, common, internal, and external carotid arteries), which supply blood to the brain. However, histopathological changes in the primate brain, caused by ischemic injury, have not previously been thoroughly investigated. In the present study, macaque monkeys were subjected to 5-18-min ischemia by occluding the eight vessels. After the brains were perfused and fixed 5 days after the occlusion, all regions were histologically investigated for ischemic cell changes. Ischemia for 5 min produced no ischemic cell change. Ischemia for 10-15 min produced cell death limited to the deeper portion of the pyramidal cell layer of the CA1 subfield in the hippocampus. In most monkeys, no cell death was observed in any brain region outside of the hippocampus after ischemia for up to 15 min. Ischemia for 18 min produced more widespread cell death in the CA1 subfield of the hippocampus, and cell death was no longer confined to the hippocampus, but was observed in layers III, V, and VI of the neocortices, the striatum, and some other regions. Brains that were perfused and fixed 1 year after 15-min ischemic insult revealed no ischemic cell morphological change in any region, but the number of pyramidal cells in the CA1 subfield was decreased to about half. The results indicate that the CA1 subfield of the monkey hippocampus is the precise region of the brain most susceptible to ischemic insult in the primate forebrain, and after a critical time (15-min ischemia in this procedure) ischemic cell changes occur suddenly and extensively. Ischemia due to occlusion of eight arteries for 10-15 min could produce a model of human amnesia caused by transient ischemic insult.

Rat preference for food-related odors.

Preferences for food-related odors and the effects of fasting on those preferences were investigated during rat bar pressing for brief odor presentation. A rat was housed in an equilateral octagonal cage and had free access to food and water, except during fasting. Among 8 food-related odor substances (black pepper, cheese, coffee, milk, nut, peppermint, plum and orange), black pepper, milk and coffee were most preferred, and cheese was least preferred, but even the bar pressing rate for cheese was above the operant level. This data indicates that all 8 odors were preferred by rats, although there were different degrees of preference in individual animals. Fasting substantially increased the rate of bar pressing for odors and changed the odors preferences. This result was probably due to increased search for food and water. Since bar pressing was reinforced by nothing other than odor presentation, the results reveal inherent odor preferences of rats.

Physiological results of monkey brain ischemia, and protection by a calcium blocker.

Physiological and histological investigation was undertaken to examine dynamic and metabolic changes due to transient ischemic insult of the monkey brain with and without postischemic treatment by the calcium entry blocker, NC-1100 (1 mg/kg, IV). Monkeys were subjected to temporary occlusion of the eight major arteries: bilateral common carotid, internal and external carotid, and vertebral arteries. Blood flow was restored after 5-, 10-, 13-, and 15-min ischemia in different monkeys. The amplitudes of extradural, cortical, and hippocampal electroencephalograms decreased severely within 1-6 min after beginning occlusion. Complete recovery of these electroencephalograms required more than 1 h. During ischemia, significant change was obvious in arterial glucose, and systolic, diastolic, and mean blood pressure, all of which increased. There were no significant physiological differences between the untreated and NC-1100-treated groups, except decreased diastolic blood pressure and slightly lower postischemic heart rate in the treated group. These small differences might be accounted for by the effect of the calcium blocker. Ten to 15 minutes ischemia caused cell changes, including cell death, which were confined almost exclusively to the CA1 subfield of untreated hippocampi examined the fifth day after occlusion. However, no ischemia-induced cell change was observed in the CA1 subfield of hippocampi subjected to 10 to 15 min ischemia in the NC-1100-treated group. It was concluded that a calcium entry blocker can protect neurons from mild ischemia-induced injury and might ameliorate morphological damage and functional impairment of the brain due to ischemia in patients who suffer transient anoxic or hypoxic injury. The present physiological data should contribute to their clinical treatment.

Calcium entry blocker ameliorates ischemic neuronal damage in monkey hippocampus.

Effects of treatment with (+/-)-1-(3,4-dimethoxyphenyl)-2-(4- diphenylmethylpiperazinyl)ethanol dihydrochloride (NC-1100), a calcium entry blocker, on ischemic neuronal damage were investigated. Monkeys were subjected to temporary occlusion of eight (bilateral common carotid, internal and external carotid, and vertebral arteries) major arteries. Blood flow was restored after 5, 10, 13, and 15 min occlusion, and NC-1100 (1 mg/kg) was then immediately infused intravenously. Monkeys were killed by perfusion fixation 5 days after occlusion. All brain regions were then histologically investigated for ischemic neuronal changes. Physiological data of NC-1100-treated subjects were not significantly different than those of untreated subjects. Heart rate tended to decrease after ischemia in treated subjects. Occlusion of 8 arteries for 10 to 15 min produced ischemic neuronal damage confined exclusively to the CA1 subfield of the hippocampus. Treatment with NC-1100 markedly reduced ischemic neuronal damage in the CA1 subfield of the hippocampus. It is suggested that postischemic treatment with the calcium entry blocker, NC-1100, might protect the brain from the ischemic damage produced in patients suffering from transient ischemia.

Hippocampal neuronal damage after transient forebrain ischemia in monkeys.

To investigate cerebral injury in the monkey due to transient ischemia, monkeys were each subjected to temporary occlusion of eight (bilateral common carotid, internal and external carotid, and vertebral) major arteries. After 0 (control), 5, 10, 13, 15, and 18 min occlusion, blood flow was restored. The monkeys were sacrificed by perfusion fixation 5 days after the operation, and all brain regions were then histologically examined for ischemic neuronal changes induced by the occlusion. The amplitude of EEG signals from skull and scalp became almost isoelectric within 1-6 min after the onset of occlusion. The EEG signals from the hippocampus were markedly attenuated within 1-4 min, although they did not become completely isoelectric. Blood pressure was significantly increased after 10-min ischemia. Five-min occlusion produced no ischemic neuronal changes except a slight increment of glial cells in the striatum and III, V, and VI layers of the neocortices. After 10- to 15-min occlusion, there were ischemic cell changes restricted exclusively to the CA1 subfield of the hippocampus. Eighteen-min occlusion produced more prominent ischemic neuronal damage in the CA1 subfield of the hippocampus, but ischemic neuronal damage was no longer confined to the hippocampus. These results suggest that only the CA1 subfield of the monkey hippocampus could be damaged by mild ischemic insult. We demonstrate that the limited lesion of the hippocampus, especially the CA1 subfield, after 10- to 15-min occlusion of eight arteries in the monkey, produces a model equivalent to human amnesia caused by transient ischemic insult.

Amino acid and NaCl appetite, and LHA neuron responses of lysine-deficient rat.

Rats' preferences for amino acids and NaCl in their drinking behavior were noted when they were fed either a control diet of gluten plus 20% purified egg protein, or (same rats, different time) a lysine-deficient diet. In control, the order of the rats' preferences was arginine greater than saline greater than monosodium L-glutamate (MSG, umami) greater than glycine greater than water greater than threonine greater than histidine greater than lysine. When fed a lysine-deficient diet the order of preference was lysine greater than saline greater than MSG greater than glycine greater than threonine greater than water greater than arginine greater than histidine. To relate neural activity with preference for amino acids and NaCl, activity of lateral hypothalamic (LHA) neurons was recorded during ingestion of MSG, lysine, arginine, glycine, saline, glucose, or water by the control and lysine-deprived condition, following a different cue tone. When the diet was lysine deficient, some neurons responded specifically to lysine ingestion. More neurons responded nondifferentially during licking in control, and responded to lysine, but fewer to other amino acid ingestion during lysine deficiency. Responses to cue tone were associated with those during licking. The present results suggest that preference for deficient amino acids might be mediated in the LHA.

Contribution of amygdalar and lateral hypothalamic neurons to visual information processing of food and nonfood in monkey.

Visual information processing was investigated in the inferotemporal cortical (ITCx)-amygdalar (AM)-lateral hypothalamic (LHA) axis which contributes to food-nonfood discrimination. Neuronal activity was recorded from monkey AM and LHA during discrimination of sensory stimuli including sight of food or nonfood. The task had four phases: control, visual, bar press, and ingestion. Of 710 AM neurons tested, 220 (31.0%) responded during visual phase: 48 to only visual stimulation, 13 (1.9%) to visual plus oral sensory stimulation, 142 (20.0%) to multimodal stimulation and 17 (2.4%) to one affectively significant item. Of 669 LHA neurons tested, 106 (15.8%) responded in the visual phase. Of 80 visual-related neurons tested systematically, 33 (41.2%) responded selectively to the sight of any object predicting the availability of reward, and 47 (58.8%) responded nondifferentially to both food and nonfood. Many of AM neuron responses were graded according to the degree of affective significance of sensory stimuli (sensory-affective association), but responses of LHA food responsive neurons did not depend on the kind of reward indicated by the sensory stimuli (stimulus-reinforcement association). Some AM and LHA food responses were modulated by extinction or reversal. Dynamic information processing in ITCx-AM-LHA axis was investigated by reversible deficits of bilateral ITCx or AM by cooling. ITCx cooling suppressed discrimination by vision responding AM neurons (8/17). AM cooling suppressed LHA responses to food (9/22). We suggest deep AM-LHA involvement in food-nonfood discrimination based on AM sensory-affective association and LHA stimulus-reinforcement association.

Magnetic resonance imaging and histologic evidence of postoperative back muscle injury in rats.

STUDY DESIGN: Postoperative back muscle injury was evaluated in rats by magnetic resonance imaging and histologic analyses. OBJECTIVE: To compare the magnetic resonance imaging manifestation of back muscle injury with the histologic findings in rats and to subsequently clarify the histopathologic appearance of the high intensity regions on T2-weighted images in human postoperative back muscles. SUMMARY OF BACKGROUND DATA: In a previous study, it was found that the signal intensity on T2-weighted images of the postoperative back muscles was increased in patients who had postsurgical lumbar muscle impairment, especially in those with a prolonged surgery duration. However, the specific histopathologic changes that cause the high signal intensity on T2-weighted images remain unclear. METHODS: Rats were divided into three groups: sham operation group, 1-hour retraction group, and 2-hour retraction group. Magnetic resonance imaging and histology of the multifidus muscles were examined before surgery and at 2, 7, and 21 days after surgery. RESULTS: T2-weighted imaging was more useful than T1-weighted imaging to estimate back muscle injury. The high signal intensity of the multifidus muscles on T2-weighted images remained 21 days after surgery only in the 2-hour retraction group. Histologically, the regeneration of the multifidus muscles was complete at 21 days after surgery in the 1-hour retraction group, but the regenerated muscle fibers in the 2-hour retraction group had a small diameter, and the extracellular fluid space remained large. CONCLUSION: The high signal intensity on T2-weighted images of the postoperative multifidus muscles in the regenerative phase may be due to an increased extracellular space and incomplete muscle fiber regeneration.

A case of a primary hepatic tumor causing segmental changes on imaging and its relation to Zahn's infarct.

A 69-year-old female was admitted to our hospital for further examination of an intrahepatic mass which had been found while undergoing a complete physical examination. The mass measured 4 cm in size and was located in the medial segment (S4) of the liver. On computed tomography (CT), S4 was observed to be 'atrophied' and was well enhanced segmentally. A celiac angiogram showed segmental staining, and a transarterial portogram demonstrated portal stoppage of S4 from the left branch. However, no segmental intensity difference was seen on magnetic resonance imaging (MRI). An aspiration biopsy showed adenocarcinoma and thus an operation was performed under a tentative diagnosis of intrahepatic cholangiocarcinoma. The postoperative diagnosis of the tumor was combined hepatocellular and cholangiocellular carcinoma. However, no histological abnormality was seen in S4, contrary to the expectation of Zahn's infarct. In this study, we discuss the mechanism and imaging findings of Zahn's infarct, the possible reasons as to why no pathological change was seen in S4, as well as stress the rarity of reports on Zahn's infarct in cases of portal thrombus due to hepatocellular carcinoma.