Enhancement of erythroid target cells for Friend murine leukemia virus by intravenous pyran treatment.

Male BALB/c mice that received prophylactic iv treatment with pyran had significantly enhanced splenomegaly, an increased number of splenic foci induced by the spleen focus forming virus (SFFV) in the Friend murine leukemia virus (F-MuLV) complex, and a slightly decreased mean survival time as compared with untreated controls infected with F-MuLV. A corresponding increase in the lymphatic leukemia virus component of the F-MuLV complex was not observed, which suggests that the enhancement of the disease was due primarily to a selective increase in the SFFV component of the F-MuLV complex. That the enhancement was related to an increased number of target cells for SFFV was substantiated by data concerning erythropoiesis in iv pyran-treated animals. Increases in splenic hematocrits and in uptake of 59Fe in the spleens of animals treated iv with pyran provided quantitative evidence for the histologic finding of increased erythroid precursors in the spleens.

Cellular and serum involvement in protection against Friend leukemia virus.

Treatment of mice with the immunomodulator pyran copolymer inhibited leukemogenesis produced by Friend leukemia virus (FLV) complex, as evidenced by inhibition of the spleen focus-forming virus and lymphatic leukemia virus, as well as by a significant decrease in splenomegaly. In this report we present data suggesting that the protective effect of pyran is mediated by macrophages. Protection was conferred on normal recipient mice when peritoneal exudate cells from pyran-treated mice were transferred to recipient mice infected 24 hr later with FLV. Animals receiving pyran-activated peritoneal cells had a significant reduction of splenomegaly and of titers of spleen focus-forming virus and lymphatic leukemia virus than did control animals. In contrast, when glycogen-elicited peritoneal exudate cells were transferred, the mice were not protected. Pyran-activated peritoneal cells, but not normal peritoneal cells, also inhibited FLV growth in vitro. Serum from pyran-treated, but not glycogen-treated, mice also transferred resistance to FLV-infected mice.

Inhibition and enhancement of Friend leukemia virus by pyran copolymer.

Inhibition or enhancement of Friend leukemia virus disease could be produced by treatment of mice with the immunopotentiator, pyran copolymer. The result depended on the route of inoculation of the drug. Prophylactic administration of the drug i.p. retarded splenomegaly, reduced splenic foci, and increased survival time of mice infected with Friend leukemia virus. Conversely, when the same dose and regimen of pyran was administered i.v., splenomegaly was enhanced, splenic foci were increased, and survival time was decreased. Histopathological examination of the spleens of mice revealed that i.p. pyran administration caused a marked increase in the splenic marginal zone with some increase in erythropoiesis in the red pulp, while i.v. pyran administration did not markedly change the splenic marginal zone but caused an early and sustained increase in erythropoiesis in the red pulp.

Stimulation of autologous antitumor T-cell responses against medullary thyroid carcinoma using tumor lysate-pulsed dendritic cells.

Dendritic cells (DCs) have attracted wide interest because of their unique capacity to elicit primary and secondary antitumor responses. We have generated autologous tumor lysate-pulsed DCs from three patients with medullary thyroid carcinoma (MTC) and tested them for their ability to stimulate cytotoxic T-cell responses against autologous MTC tumor cells in vitro. The aim of our investigations was to evaluate the potential efficacy of DC-based immunotherapy in patients with MTC. DCs were generated from peripheral blood monocytes using GM-CSF and IL-4 (immature DCs) or GM-CSF, IL-4, and TNFalpha (mature DCs). Our results indicate that mature tumor lysate-pulsed DCs are able to elicit a human leukocyte antigen class I-restricted cytotoxic T-cell response against autologous MTC tumor cells, whereas immature tumor lysate-pulsed DCs do not stimulate significant antitumor activity. We feel that our data may be relevant for future clinical trials of active immunotherapy using tumor lysate-pulsed DCs in patients with MTC who have residual or distant disease after surgical treatment. The fact that mature DCs displayed a substantially higher capacity to stimulate autologous antitumor T-cell responses than immature DCs underlines the importance of a maturation step in immunotherapy protocols based on DCs.

Facilitation and Delay Sensitivity of Auditory Cortex Neurons in CF - FM Bats, Rhinolophus rouxi and Pteronotus p.parnellii.

Responses of auditory neurons to complex stimuli were recorded in the dorsal belt region of the auditory cortex of two taxonomically unrelated bat species, Rhinolophus rouxi and Pteronotus parnellii parnellii, both showing Doppler shift compensation behaviour. As in P.p.parnellii (Suga et al., J. Neurophysiol., 49, 1573 - 1626, 1983), cortical neurons of R.rouxi show facilitated responses to pairs of pure tones or frequency modulations. Best frequencies for the two components lie near the first and second harmonic of the echolocation call but are in most cases not harmonically related. Neurons facilitated by pairs of pure tones show little dependence on the delay between the stimuli, whereas pairs of frequency modulations evoke best facilitated responses at distinct best delays between 1 and 10 ms. Facilitated neurons are found in distinct portions of the dorsal cortical belt region, with a segregation of facilitated neurons responding to pure tones and to frequency modulations. Non-facilitated neurons are found throughout the field. Neurons are topographically aligned with increasing best delays along a rostrocaudal axis. The best delays between 2 and 4 ms are largely overrepresented numerically, and occupy approximately 56% of the cortical area containing facilitated neurons. A functional interpretation of the large overrepresentation of best delays approximately 3 ms is proposed. Facilitated neurons are located almost entirely within layer V of the dorsal field.

Anatomy and projection patterns of the superior olivary complex in the Mexican free-tailed bat, Tadarida brasiliensis mexicana.

The superior olivary complex (SOC) is the first station in the ascending auditory pathway that receives binaural projections. Two of the principal nuclei, the lateral superior olive (LSO) and the medial superior olive (MSO), are major sources of ascending projections to the inferior colliculus. Whereas almost all mammals have an LSO, it has traditionally been thought that only animals that hear low frequencies have an MSO. Recent reports, however, suggest that the medial part of the SOC in bats is highly variable and that at least some bats have a well-developed MSO. Thus, the main goal of this study was to evaluate the cytoarchitecture and connections of the principal superior olivary nuclei of the Mexican free-tailed bat, with specific attention directed at the MSO. Cell and fiber stained material revealed that the LSO and the medial nucleus of the trapezoid body (MNTB) are similar to those described for other mammals. There are two medial nuclei we refer to as dorsomedial periolivary nucleus (DMPO) and MSO. Tracer experiments exhibited that the DMPO receives bilateral projections from the cochlear nucleus, and additional projections from the ipsilateral MNTB. The DMPO sends a strong projection to the ipsilateral inferior colliculus. Positive staining for acetylcholinesterase indicates that the DMPO is a part of the olivocochlear system, as it is in other animals. The MSO in the free-tailed bat meets many of the criteria that traditionally define this nucleus. These include the presence of bipolar and multipolar principal cells, bilateral innervation from the cochlear nucleus, a strong projection from the ipsilateral MNTB, and the absence of cholinergic cells. The major difference from traditional MSO features is that it projects bilaterally to the inferior colliculus. Approximately 30% of its cells provide collateral projections to the colliculi on both sides. Functional implications of the MSO for the free-tailed bat are considered in the Discussion.

The bovine papillomavirus type 1 genome contains multiple loci of static DNA bending, but bends are absent from the functional origin of replication.

Twenty-four overlapping restriction fragments spanning the entire bovine papillomavirus type 1 (BPV-1) genome were analyzed by electrophoresis to determine the extent of static DNA bending in the BPV-1 genome. Thirteen of 24 fragments contained static bends. Based on known locations of previously mapped bend loci and the overlapping pattern of these 13 fragments, we estimate that there are 8-11 distinct static bend loci in the BPV-1 genome. The bend loci were not uniformly distributed on the genome and with one exception, were clustered from nucleotides 5816 to 2621 on the BPV-1 map. This portion of the BPV-1 genome contains most of the transcriptional regulatory sequences as well as the origin of replication. The concordance between the genomic distribution of DNA bends and cis-active elements is consistent with the possibility that bent sequences may contribute to the function of at least some of these elements. However, unlike SV40, there was no static bend at that functional origin of replication for BPV-1. The nearest bends to the origin were approximately 120 bp to the 5' side and 300 bp to the 3' side. As both of these bends were outside of the sequences required for origin function, it is unlikely that static bending plays a critical role in BPV-1 replication.

A cheap earphone for small animals with good frequency response in the ultrasonic frequency range.

A new type of earphone for binaural stimulation in small animals is described. The characteristic features are: (1) small, 8 mm diameter, can be miniaturised further; (2) cheap, uses no expensive condenser microphone capsule, material costs less than 20 dollars; (3) frequency response flat within 6 dB between 10 kHz and 120 kHz, within 10 dB up to 200 kHz; (4) harmonic distortions at levels of the fundamental frequency below 75 dB SPL is below - 34 dB at worst case (10 kHz); and (5) due to the low price the most stable earphones optimally paired for dichotic stimulation can be selected by screening a number of individual specimen.

A stereotaxic method for small animals using experimentally determined reference profiles.

In bats conventional stereotaxic methods do not yield sufficient positional accuracy to allow reliable recordings and tracer injections in subnuclei of the auditory system. In a newly developed stereotaxic system experimentally measured patterns of skull profile lines are used to define the animal's brain position with an accuracy of +/- 100 microns. By combining the neurophysiological stereotaxic procedure with a standardization of the neuroanatomical processing of the brains, the location of recordings, stimulations or injections can be readily transformed into brain atlas coordinates. This facilitates the compilation and comparison of data within and among animals. The system is not restricted to use in bats and can be readily adapted to other experimental animals.

An open, modular, distributed and redundant computer system for hospitals.

Based on his experience at the University of Würzburg in Germany, Dr. Schuller shows how a successful MUMPS-based hospital information system can be developed, including goals and requirements, networking, software hierarchies, programming in MUMPS, medical applications, data security/privacy and international standards.

Coding of small sinusoidal frequency and amplitude modulations in the inferior colliculus of 'CF-FM' bat, Rhinolophus ferrumequinum.

Single neurons in the inferior colliculus of the Greater Horseshoe bat, Rhinolophus ferrumequinum, showed two broad categories of response patterns to sinusoidally frequency (SFM) or amplitude (SAM) modulated stimuli. Tonic responding cells (best excitatory frequency (BEF) between 10 and 90 kHz) showed a rough sinusoidal modulation of the discharge pattern to SFM. Transient responding neurons generally showing on- or off-responses to pure tones, (BEF between 65 and 88 kHz), displayed highly synchronized discharge patterns to SFM-cycles (Fig. 1). Modulation rates between 20 and 100 Hz were most effective and some neurons encoded modulation rates up to 350 Hz (Figs. 2 and 3). The SFM responses were best synchronized to the modulation envelope for center frequencies in the upper portion of the tuning curve (Figs. 4 and 5). Sharply tuned neurons with BEF around 80 kHz had the lowest threshold for modulation depth (+/- 10 Hz or 0.025%) (Fig. 6). In general, SAMs evoked the same type of response patterns and were encoded down to modulation index of 3% (Fig. 7). The fine frequency and amplitude discriminations for periodical modulations by collicular neurons is discussed as related to the detection and discrimination performance of bats, when preying on flying insects in clustered surroundings.

Spectral and temporal gating mechanisms enhance the clutter rejection in the echolocating bat, Rhinolophus rouxi.

Doppler shift compensation behaviour in horseshoe bats, Rhinolophus rouxi, was used to test the interference of pure tones and narrow band noise with compensation performance. The distortions in Doppler shift compensation to sinusoidally frequency shifted echoes (modulation frequency: 0.1 Hz, maximum frequency shift: 3 kHz) consisted of a reduced compensation amplitude and/or a shift of the emitted frequency to lower frequencies (Fig 2). Pure tones at frequencies between 200 and 900 Hz above the bat's resting frequency (RF) disturbed the Doppler shift compensation, with a maximum of interference between 400 and 550 Hz (Fig. 1). Minimum duration of pure tones for interference was 20 ms and durations above 40 ms were most effective (Fig. 3). Interfering pure tones arriving later than about 10 ms after the onset of the echolocation call showed markedly reduced interference (Fig. 4). Doppler shift compensation was affected by pure tones at the optimum interfering frequency with sound pressure levels down to -48 dB rel the intensity level of the emitted call (Fig. 5, 6). Narrow bandwidth noise (bandwidth from +/- 100 Hz to +/- 800 Hz) disturbed Doppler shift compensation at carrier frequencies between -250 Hz below and 800 Hz above RF with a maximum of interference between 250 and 500 Hz above resting frequency (Fig. 7). The duration and delay of the noise had similar influences on interference with Doppler shift compensation as did pure tones (Fig. 8, 9). Intensity dependence for noise interference was more variable than for pure tones (-32 dB to -45 dB rel emitted sound pressure level Fig. 10).(ABSTRACT TRUNCATED AT 250 WORDS)

Motion processing in the auditory cortex of the rufous horseshoe bat: role of GABAergic inhibition.

This study examined the influence of inhibition on motion-direction-sensitive responses of neurons in the dorsal fields of auditory cortex of the rufous horseshoe bat. Responses to auditory apparent motion stimuli were recorded extracellularly from neurons while microiontophoretically applying gamma-aminobutyric acid (GABA) and the GABAA receptor antagonist bicuculline methiodide (BMI). Neurons could respond with a directional preference exhibiting stronger responses to one direction of motion or a shift of receptive field (RF) borders depending on direction of motion. BMI influenced the motion direction sensitivity of 53% of neurons. In 21% of neurons the motion-direction sensitivity was decreased by BMI by decreasing either directional preference or RF shift. In neurons with a directional preference, BMI increased the spike number for the preferred direction by a similar amount as for the nonpreferred direction. Thus, inhibition was not direction specific. BMI increased motion-direction sensitivity by either increasing directional preference or magnitude of RF shifts in 22% of neurons. Ten percent of neurons changed their response from a RF shift to a directional preference under BMI. In these neurons, the observed effects could often be better explained by adaptation of excitation rather than inhibition. The results suggest, that adaptation of excitation, as well as cortex specific GABAergic inhibition, contribute to motion-direction sensitivity in the auditory cortex of the rufous horseshoe bat.

Cortical representation of acoustic motion in the rufous horseshoe bat, Rhinolophus rouxi.

Responses of neurons to apparent auditory motion in the azimuth were recorded in three different fields of auditory cortex of the rufous horseshoe bat. Motion was simulated using successive stimuli with dynamically changing interaural intensity differences presented via earphones. Seventy-one percent of sampled neurons were motion-direction-sensitive. Two types of responses could be distinguished. Thirty-four percent of neurons showed a directional preference exhibiting stronger responses to one direction of motion. Fifty-seven percent of neurons responded with a shift of spatial receptive field position depending on direction of motion. Both effects could occur in the same neuron depending on the parameters of apparent motion. Most neurons with contralateral receptive fields exhibited directional preference only with motion entering the receptive field from the opposite direction. Receptive field shifts were opposite to the direction of motion. Specific combinations of spatiotemporal parameters determined the motion-direction-sensitive responses. Velocity was not encoded as a specific parameter. Temporal parameters of motion and azimuth position of the moving sound source were differentially encoded by neurons in different fields of auditory cortex. Neurons with a directional preference in the dorsal fields can encode motion with short interpulse intervals, whereas direction-preferring neurons in the primary field can best encode motion with medium interpulse intervals. Furthermore, neurons with a directional preference in the dorsal fields are specialized for encoding motion in the midfield of azimuth, whereas direction-preferring neurons in the primary field can encode motion in lateral positions. The results suggest that motion information is differentially processed in different fields of the auditory cortex of the rufous horseshoe bat.

Periaqueductal gray and the region of the paralemniscal area have different functions in the control of vocalization in the neotropical bat, Phyllostomus discolor.

The periaqueductal gray matter and the region of the paralemniscal area were neuroanatomically delineated in the brain of the neotropical bat Phyllostomus discolor[Wagner (1843) Arch. Naturgesch., 9, 365-368] and were probed with electrical microstimulation for eliciting vocalizations. In a well-delimited rostral portion of the periaqueductal gray exclusively, communication calls could be triggered at low stimulation currents. Communication calls as well as echolocation calls could be elicited at the dorsal and ventral edges of this area. Pharmacological stimulation with microdialysed kainic acid in this particular periaqueductal gray area demonstrated that neurons and not fibres of passage are activated for triggering vocalization. Solely echolocation calls were emitted upon electrical microstimulation or with microdialysed kainic acid in the region of the paralemniscal area. The periaqueductal gray appears to be involved in vocal pathways that control both communication calls and echolocation calls, while the region of the paralemniscal area seems to be specialized for control of echolocation calls only. Respiration is similarly influenced by stimulation in the periaqueductal gray and the region of the paralemniscal area. Periaqueductal gray and paralemniscal area interact differently with the final common pathway for vocalization, and may represent different functional organization in the vocal controlling pathways for communication calls and echolocation calls.

The central acoustic tract and audio-vocal coupling in the horseshoe bat, Rhinolophus rouxi.

Doppler shift compensation (DSC) behaviour in horseshoe bats is a remarkable example of sensorimotor feedback that stabilizes the echo frequency at the bat's optimum hearing range regardless of motion-induced frequency shifts in the echoes. Searching for a related neural interface, the nucleus of the central acoustic tract (NCAT) was investigated in the echolocating horseshoe bat, Rhinolophus rouxi, using various neurophysiological and tracer methods. The NCAT receives bilateral auditory input from the cochlear nuclei and sends projections to regions outside the classical acoustic pathway like the pretectal area or the superior colliculus. The binaural input is excitatory from the contralateral and inhibitory from the ipsilateral ear to 53% of the units, and auditory responses were biased to frontal and contralateral directions. The best frequencies of NCAT neurons match a narrow range above the main frequency component of the bat's species-specific echolocation call (62% of the units), and the neurons exhibit extremely sharp tuning (Q10dB up to 632). DSC is degraded by unilateral electrical or pharmacological microstimulation of the NCAT, and heavily impaired by unilateral lesion of the region. Altogether, the efferents of the NCAT to prevocal areas, the tuning of its neurons to the DSC-relevant echo frequency range, and the possibility to affect DSC by manipulation of the NCAT, support the assumption that the nucleus plays an important role in audio-vocal control in the horseshoe bat.

Audiovocal behavior of Doppler-shift compensation in the horseshoe bat survives bilateral lesion of the paralemniscal tegmental area.

The role of the paralemniscal tegmental area of the horseshoe bat, Rhinolophus rouxi, in the control of vocalization and Doppler-shift compensation was investigated using electrical and pharmacological stimulation and lesioning techniques. The paralemniscal tegmental area is situated in the dorsolateral tegmentum ventral to the inferior colliculus and rostral and medial to the dorsal and intermediate nuclei of the lateral lemniscus. Vocalizations indistinguishable from spontaneously uttered calls can be elicited with both electrical and pharmacological stimulation methods, demonstrating that the stimulation of neural elements within the area and not fibers passing through the area are responsible for the stimulated call emission. The audiovocal feedback system for Doppler-shift compensation was also investigated. Doppler-shift compensation adjusts the frequency of the emitted calls according to the increases in the frequency of the echoes that are normally encountered in flying bats. Bats compensate for Doppler shifts not only under natural conditions but also when echoes are played back to the bat following spontaneous vocalizations or vocalizations induced by electrical or pharmacological stimulation of the investigated brain area. Unilateral electrolytic lesions of the paralemniscal tegmental area did not impair the ability to evoke vocalizations with electrical stimulation of the unlesioned side. The calls had exactly the same structure and frequency composition as those emitted prior to lesioning. Unilateral lesions also did not impair Doppler-shift compensation performance. After bilateral lesioning of the paralemniscal area, vocalizations could not be evoked with electrical stimulation. However, normal calls were emitted spontaneously and Doppler-shift compensation during spontaneous call emission was unaltered compared with the intact condition. The paralemniscal tegmental area is therefore not an audiovocal feedback system required for Doppler-shift compensation, but rather a brain area whose stimulation and activation is sufficient but not necessary for call emission. It is also not directly involved in the control of spectral parameters of vocalization but contributes to the control of the occurrence of vocal output.

Auditory cortex of the rufous horseshoe bat: 1. Physiological response properties to acoustic stimuli and vocalizations and the topographical distribution of neurons.

The extent and functional subdivisions of the auditory cortex in the echolocating horseshoe bat, Rhinolophus rouxi, were neurophysiologically investigated and compared to neuroarchitectural boundaries and projection fields from connectional investigations. The primary auditory field shows clear tonotopic organization with best frequencies increasing in the caudorostral direction. The frequencies near the bat's resting frequency are largely over-represented, occupying six to 12 times more neural space per kHz than in the lower frequency range. Adjacent to the rostral high-frequency portion of the primary cortical field, a second tonotopically organized field extends dorsally with decreasing best frequencies. Because of the reversed tonotopic gradient and the consistent responses of the neurons, the field is comparable to the anterior auditory field in other mammals. A third tonotopic trend for medium and low best frequencies is found dorsal to the caudal primary field. This area is considered to correspond to the dorsoposterior field in other mammals. Cortical neurons had different response properties and often preferences for distinct stimulus types. Narrowly tuned neurons (Q10dB > 20) were found in the rostral portion of the primary field, the anterior auditory field and in the posterior dorsal field. Neurons with double-peaked tuning curves were absent in the primary area, but occurred throughout the dorsal fields. Vocalization elicited most effectively neurons in the anterior auditory field. Exclusive response to pure tones was found in neurons of the rostral dorsal field. Neurons preferring sinusoidal frequency modulations were located in the primary field and the anterior and posterior dorsal fields adjacent to the primary area. Linear frequency modulations optimally activated only neurons of the dorsal part of the dorsal field. Noise-selective neurons were found in the dorsal fields bordering the primary area and the extreme caudal edge of the primary field. The data provide a survey of the functional organization of the horseshoe bat's auditory cortex in real coordinates with the support of cytoarchitectural boundaries and connectional data.

Significance of the paralemniscal tegmental area for audio-motor control in the moustached bat, Pteronotus p. parnellii: the afferent off efferent connections of the paralemniscal area.

The paralemniscal tegmental area has been determined in the brain of the New World moustached bat, Pteronotus p. parnellii, by electrical microstimulation eliciting echolocation calls and pinna movements. It is located in the dorsal tegmentum rostral and medial to the dorsal nucleus of the lateral lemniscus and is characterized by medium sized and large neurons. Tracer injections (WGA-HRP) showed that the most intense input to the paralemniscal tegmental area originates in the intermediate and deep layers of the homolateral superior colliculus. The strong projections from the ipsi- and contralateral nucleus praepositus hypoglossus most probably contributes vestibular information. Further inputs in descending order of intensity are from the substantia nigra, the contralateral paralemniscal tegmental area, the putamen, the ventral reticular formation in its lateral portions, the medial cerebellar nucleus and the dorsal reticular formation. Efferent projections of the paralemniscal tegmental area reach the putamen bilaterally, the nucleus accumbens and other parts of the basal ganglia, the pretectal area, the substantia nigra, the intermediate and deep layers of the superior colliculus bilaterally and the tegmental area ventral to it. Connections to the dorsal part of the periaqueductal grey, the cuneiform nucleus and the parabrachial region are important in the context of vocal control, whereas projections to the medial portion of the contralateral facial nucleus may interfere with the control of pinna movement. The findings suggest that the paralemniscal tegmental area is involved in audio-motor control of vocalization and pinna movements in bats; connectional and functional similarities and disparities to tegmental regions described in other mammals are discussed.