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Hydrocortisone inhibits granulocyte-macrophage colony-stimulating factor production from normal human peripheral blood mononuclear cells and CD3+ T cells.

Normal human peripheral blood mononuclear cells (MNCs), particularly T lymphocytes (T cells), are a rich source of granulocyte-macrophage colony-stimulating factor (GM-CSF). Glucocorticoids are known to inhibit GM-CSF production in in vitro cultures of a human fibroblast cell line and in normal human blood monocytes and alveolar macrophages. To determine whether glucocorticoids also inhibit GM-CSF production from normal human MNCs and T cells, we set up cultures of normal human MNCs and T cells in a liquid system in the presence and absence of 5, 50, and 250 microg/dL of hydrocortisone, and an hour later, a constant dose of 50-ng/mL Escherichia coli lipopolysaccharide (LPS) or 10-microg/mL phytohemagglutinin (PHA) was added. After three days, cell counts and GM-CSF levels were determined. Administering 50- and 250-microg/dL hydrocortisone decreased lymphocyte recovery from MNC cultures with LPS (p < or = 0.01), and 250 microg/dL of hydrocortisone decreased lymphocyte recovery from MNC and T-cell cultures with PHA (p < or = 0.03). The amount of GM-CSF produced from PHA-stimulated MNCs was about 100-fold higher than that produced from LPS-stimulated MNCs. The magnitude of GM-CSFs produced in MNC and T-cell cultures stimulated by PHA was comparable (p=0.88). Administering hydrocortisone at 5, 50, and 250 pg/dL decreased GM-CSF production (p < 0.003) in LPS- or PHA-stimulated MNC cultures and in PHA-stimulated T-cell cultures. PHA (not tested with LPS)-stimulated GM-CSF messenger RNA (mRNA) expression was blocked by hydrocortisone. These results indicate that lower concentrations of hydrocortisone inhibit GM-CSF production from normal human blood MNCs and T cells entirely by inhibiting the expression of GM-CSF mRNA, and higher concentrations of hydrocortisone inhibit by a combined effect of inhibiting the expression of GM-CSF mRNA and decreasing the lymphocyte count.

Apoptosis↗

Holter monitor recording for the detection of myocardial ischemia: validation of a new recorder and chest lead positions.

OBJECTIVE: To validate the Reynolds Tracker II Holter system using newly described lead positions during both upright treadmill exercise and in the recumbent position following exercise. The specific lead positions HL1 and HL2 were chosen to detect anterior and postero-inferior myocardial ischemia, respectively, without interfering with the surgical field in the hypothetical situation of open-heart surgery. Similar lead positions have previously been used to monitor myocardial ischemia during induction of anesthesia, but have never been validated by comparison with 12-lead modified electrocardiogram (ECG) recording. METHODS: To validate the authors' 'chosen' Holter lead positions (HL1 and HL2), both at the fifth intercostal space just lateral to the midclavicular line and on the back, 1.5 cm to the left of the vertebral column, respectively) 49 candidates for routine treadmill exercise testing underwent a simultaneous Holter monitor recording using the described lead positions. DATA ANALYSIS: The Holter ECG recordings were separately analyzed by two physicians unaware of patients' identity. RESULTS: Using the modified 12-lead ECG as the 'gold standard', the sensitivity of Holter for detecting ischemia (defined as 0.1 mV or ST depression lasting at least 60 s) was 77 and 83%, and its specificity was 100 and 92%, respectively, for observers 1 and 2. Most episodes of myocardial ischemia were detected by the modified lead V5 for the 12-lead ECG and by HL1 for the Holter recording. Using the Holter Tracker II system and the chosen lead positions, it was possible to detect successfully most episodes of exercise-induced myocardial ischemia. CONCLUSION: Holter monitoring might be useful in detecting perioperative myocardial ischemia.

Coronary Disease↗

Topographic organization of the orientation column system in large flat-mounts of the cat visual cortex: a 2-deoxyglucose study.

We developed a flat-mount technique in order to visualize, without additional reconstruction, the system of orientation columns in the cat visual cortex by using 2-deoxyglucose-autoradiography. Experimental animals were injected with 2-deoxyglucose and then stimulated for 45-60 minutes either with vertical or horizontal or oblique gratings alone or with vertical and horizontal gratings presented in alternation. In both areas 17 and 18 stimulation with either vertical or horizontal or oblique stripes produced similar and highly ordered patterns of parallel bands of increased 2-deoxyglucose uptake that were perpendicular to the boundaries of the areas. In area 17 they occasionally extended without interruption from the 17/18 border on the top of the lateral gyrus to the monocular segment in the splenial sulcus. Superposition of serial sections revealed that these bands were present in all cortical layers and in precise register along lines orthogonal to the lamination. The center-to-center spacing of the bands was 1.0-1.1 mm in area 17 and 1.2-1.4 mm in area 18. Stimulation with alternating vertical and horizontal contours led to a pattern the general organization of which resembled that induced by a single orientation but the spacing of which was reduced by a factor of 0.5. This strongly supports the concept that orientation is mapped in a system of parallel bands and argues against a recently formulated hypothesis that iso-orientation bands extend like spokes from centers that lack orientation selectivity (Braitenberg and Braitenberg, Biol. Cybern. 33:179-186, '79). Another characteristic feature, revealed by the flat-mount technique, was a periodic variation of 2-deoxyglucose uptake along the bands that gave them a beaded appearance. The mean center-to-center distance between adjacent beads on the same band was in the range of 0.9-1.2 mm and remained unchanged when horizontal and vertical gratings were presented in alternation. We propose that these beads reflect another columnar system whose features have yet to be determined.

Animals↗

HPA-related CSF neuropeptides in suicide attempters.

Corticotropin-releasing hormone (CRH), somatostatin (SOM), delta-sleep-inducing peptide (DSIP), neuropeptide Y (NPY), beta-endorphin (beta-END), and vasopressin (AVP), which are regarded as being involved in the HPA-regulation were investigated in lumbar CSF of 44 suicide attempters. The patients were diagnosed according to the DSM-III-R, and rated with the MADRS. The neuropeptides were compared with the serotonin metabolite 5-hydroxyindoleacetic acid (5-HIAA) in CSF and with post-dexamethasone plasma cortisol. We found strong correlations between CRH and the peptides SOM and beta-END. The latter also correlated positively with SOM. There were no differences between men and women. Patients with major depressive disorders had significantly lower SOM, CRH, and DSIP than other patients. Both SOM and beta-END correlated negatively with post dexamethasone plasma cortisol in all patients. We found no significant relationships between neuropeptides and CSF 5-HIAA. Patients who had made previous suicide attempts had significantly lower CRH than those who had not. No other significant associations between neuropeptides and suicidal subgroups of patients appeared, and there was no indication of specific neuropeptide patterns in patients who later completed suicide. Intercorrelations of some neuropeptides and low SOM and DSIP in major depressed patients are findings in line with those by others.

Adult↗

[Changes in the level of sister chromatid exchanges in cultured Chinese hamster cells undergoing limited proliferation].

"Stationary phase ageing" of cultured Chinese hamster cells (when proliferation rate decreases and in the stationary growth phase) produces an increase in the frequency of spontaneous sister chromatid exchanges (SCE). Thiophosphamide-induced (24 h) frequency of SCE increases from 2-day to 5-day "age" and later (in the stationary phase) is practically the same. The "stationary ageing" cultured cells are suggested to be used as a model system for studying molecular-genetic age changes.

Animals↗

Muscle fibre direction of longissimus, iliocostalis and multifidus: landmark-derived reference lines.

Considerable inter-individual variations in the fibre direction angles of the iliocostalis lumborum, longissimus and multifidus were observed, thus bringing the applicability of a two dimensional fixed angle grid system for fibre direction determination into question. However, the angulation of the fibres of the multifidus and iliocostalis lumborum were found to be easily identifiable by the use of three surface anatomical landmarks: the caudal tip of the superior iliac spine, the lateral border of the iliocostalis at the twelfth rib and the L1-L2 interspinous space. No reliable index was found for the longissimus. Suggested electrode placement sites for the electromyographic study of the iliocostalis lumborum and the multifidus are at the levels of the L2-L3 and the L4-L5 interspinous spaces respectively.

Biometry↗

Localization of multiple sound sources with two microphones.

This paper presents a two-microphone technique for localization of multiple sound sources. Its fundamental structure is adopted from a binaural signal-processing scheme employed in biological systems for the localization of sources using interaural time differences (ITD). The two input signals are transformed to the frequency domain and analyzed for coincidences along left/right-channel delay-line pairs. The coincidence information is enhanced by a nonlinear operation followed by a temporal integration. The azimuths of the sound sources are estimated by integrating the coincidence locations across the broadband of frequencies in speech signals (the "direct" method). Further improvement is achieved by using a novel "stencil" filter pattern recognition procedure. This includes coincidences due to phase delays of greater than 2pi, which are generally regarded as ambiguous information. It is demonstrated that the stencil method can greatly enhance localization of lateral sources over the direct method. Also discussed and analyzed are two limitations involved in both methods, namely missed and artifactual sound sources. Anechoic chamber tests as well as computer simulation experiments showed that the signal-processing system generally worked well in detecting the spatial azimuths of four or six simultaneously competing sound sources.

Adult↗

Hypothesis; overdistention of the neural tube may cause anomalies of non-neural organs.

This hypothesis is offered by a neurological surgeon interested in anomalies of the central nervous system. It is based on accumulating evidence indicating that some neural tube defects result not from failure of the tube to close but from its rupture after closure. The central nervous system, serving all organs, is the first to develop and its maldevelopment may cause damage to other emerging structures. The neural tube closes during the fourth week and is immediately distended by a proteinaceous neural tube fluid (NTF) secreted by its lining cells at a pressure four to five times that of the surrounding amniotic fluid. This NTF has been miscalled "cerebrospinal fluid." The choroid plexus does not begin to secrete true cerebrospinal fluid (CSF) until 2 weeks later. If oversecretion of NTF should occur during this 2-week interval, the resulting overexpansion of the neural tube may spread apart the developing somite, eventuating in a combination of anterior and posterior spina bifida that constitutes bilateral hemivertebrae. If the distending neural tube ruptures beneath intact cutaneous ectoderm, the escaping NTF will infiltrate mesoderm. The resulting dislocation of cells and their possible injury by the extraneous protein may damage the as yet unidentifiable anlagen of mesodermal organs. If neural tube overdistention splits the underlying notochord and damages primitive gut, anomalies of entodermal organs may result. The neuroenteric cyst is one such anomaly that the neurosurgeon is called upon to treat. He finds it accompanied by hemivertebrae and hydromyelia. A preliminary report on this hypothesis has been published (Gardner and Breuer, '80).

Central Nervous System↗

Complementation of the xeroderma pigmentosum DNA repair synthesis defect with Escherichia coli UvrABC proteins in a cell-free system.

A newly developed cell-free system was used to study DNA repair synthesis carried out by extracts from human cell lines in vitro. Extracts from a normal human lymphoid cell line and from cell lines established from individuals with hereditary dysplastic nevus syndrome perform damage-dependent repair synthesis in plasmid DNA treated with cis- or trans-diamminedichloro-platinum(II) or irradiated with ultraviolet light. Cell extracts of xeroderma pigmentosum origin (complementation groups A, C, D, and G) are deficient in DNA repair synthesis. When damaged plasmid DNA was pretreated with purified Escherichia coli UvrABC proteins, xeroderma pigmentosum cell extracts were able to carry out DNA repair synthesis. The ability of E. coli UvrABC proteins to complement xeroderma pigmentosum cell extracts indicates that the extracts are deficient in incision, but can carry out later steps of repair. Thus the in vitro system provides results that are in agreement with the incision defect found from studies of xeroderma pigmentosum cells.

Cell-Free System↗

Innervation of the ring gland of Drosophila melanogaster.

In insects, peptidergic neurons of the central nervous system regulate the synthesis of the main developmental hormones. Neuropeptides involved in this neuroendocrine cascade have been identified in lepidopterans and dictyopterans. Since these organisms are not suitable for genetic research, we identified peptidergic brain neurons innervating the ring gland in Drosophila melanogaster. In larvae of Drosophila, ecdysteroids and juvenile hormones are produced by the ring gland, which is composed of the prothoracic gland, the corpus allatum, and the corpora cardiaca. Using the GAL4 enhancer trap system, we mapped those neurons of the central nervous system that innervate the ring gland. Eleven groups of neurosecretory neurons and their target tissues were identified. Five neurons of the lateral protocerebrum directly innervate the prothoracic gland or corpus allatum cells of the ring gland and are believed to regulate ecdysteroid and juvenile hormone titers. Axons of the circadian pacemaker neurons project onto dendritic fields of these five neurons. This connection might be the neuronal substrate of the circadian rhythms of molting and metamorphosis in Drosophila. Most of the neurons presented here have not been described before. The enhancer trap lines labeling them will be valuable tools for the analysis of neuronal as well as genetic regulation in insect development.

Animals↗

Continuous monitoring of ventilatory mechanics during one-lung ventilation.

OBJECTIVE: The Ultima SV respiratory monitor can be used to monitor the intraoperative effects of the lateral decubitus position and one-lung ventilation on ventilatory mechanics. METHODS: Eight patients with esophageal cancer who required one-lung ventilation for esophagectomy and reconstruction were enrolled in the study. We monitored pressure-volume or flow-rate-volume loops continuously throughout the operation. Respiratory parameters were evaluated closely during five conditions of ventilation: two-lung ventilation in the supine position, two-lung ventilation in the lateral decubitus position, dependent one-lung ventilation in the lateral decubitus position, nondependent one-lung ventilation in the lateral decubitus position, and dependent one-lung ventilation in the lateral decubitus position with the chest opened. Respiratory rate was controlled at 10 breaths/min, and tidal volume was kept constant (10 ml/kg) during surgery. RESULTS: Peak inspiratory pressure increased to 29.0 +/- 9.0 (mean +/- SD) cm H2O in the dependent one-lung in the lateral decubitus position with the chest opened (p < 0.01). Dynamic compliance decreased to 29.4 +/- 4.9 ml/cm H2O in the dependent one-lung in the lateral decubitus position with the chest opened (p < 0.01). The changing configuration of the loops also offered additional and instantaneous information during one-lung ventilation. CONCLUSIONS: One-lung ventilation caused several changes in the whole respiratory system (lung, thorax, and endotracheal tube). Continuous monitoring of flow-rate-volume or pressure-volume loops with in-line spirometry provided comprehensive information regarding parameters in one-lung ventilation.

Aged↗

Adverse male reproductive effects following subchronic exposure of rats to sodium dichloroacetate.

Dichloroacetate (DCA) activates the pyruvate dehydrogenase complex enhancing carbohydrate and lactate utilization in animals. As a result it is used clinically in the treatment of acute lactic acidosis and has therapeutic potential in the treatment of stroke. Adverse effects of chronic DCA treatment include polyneuropathy and testicular degeneration. Since DCA is a principal product of the aqueous chlorination of fulvic acids concern has arisen regarding the agent's impact on environmental health. We treated male Long-Evans rats with 0, 31.25, 62.5, or 125 mg DCA/kg/day by oral gavage for 10 weeks. Compared to controls, preputial gland and epididymis weights were reduced at 31.25 mg/kg, body and liver weights at 62.5 mg/kg, and accessory organ weights at 125 mg/kg. Epididymal sperm counts were reduced and sperm morphology was impacted at the 62.5 and 125 mg/kg doses levels. Histologic examination of the testis and epididymis revealed inhibited spermiation in testes at the 125 mg/kg dose level. Computer-assisted sperm motion analysis revealed reductions in percentage motile sperm, curvilinear and straight-line velocity, linearity, and amplitude of lateral head displacement at both the 62.5 and the 125 mg/kg dose levels. In the assessment of fertility after an overnight mating, the number of viable implants on Day 14 of gestation was decreased only in the highest dose group. These studies demonstrate adverse effects of NaDCA treatment on the rat male reproductive system, primarily on the accessory organs and sperm within them at lower doses (31.25 and 62.5 mg/kg), and on the testis at the highest dose (125 mg/kg).

Animals↗

Experimental velocity profiles and volumetric flow via two-dimensional speckle tracking.

The performance of a two-dimensional speckle tracking system in measuring in vitro laminar flow is evaluated. The system uses a pattern matching algorithm to track subresolution-sized speckle regions between successive ultrasonic 2D pulse-echo acquisitions in order to determine both the axial and the lateral components of velocity. In this study, multiple 2D vector velocity maps were acquired in real time using a calibrated laminar flow phantom, and then statistically analyzed off-line. At a 90 degrees transducer angle, volumetric flow rates computed from measured velocity profiles exhibited excellent linearity (R2 > 0.99), with a mean error of -6.1%, over the range 5-30 mL/s. At 105 degrees and 120 degrees, experimental volume flow rates also agreed well with actual rates, although measured velocity profiles appeared more irregular with decreasing Doppler angles. Velocity profiles estimated using sampled radio-frequency data rather than envelope-detected data were inconsistent due to an insufficient sampling rate and the quantization of the velocity grid. Results indicate that excellent flow velocity and volume rate estimates can be obtained from vector velocity measurements along a single line of sight, without a priori knowledge of the flow direction, at transducer angles near 90 degrees where Doppler instruments are prone to large errors.

Phantoms, Imaging↗

Transgenic mice expressing beta-galactosidase in mature neurons under neuron-specific enolase promoter control.

To gain insights into transcription factors defining neuronal identity, we generated transgenic mice carrying a 1.8 kb rat neuron-specific enolase (NSE) promoter fragment fused to an E. coli lacZ gene. Four of seven transgenic families expressed transgene RNA in the nervous system but not in most other tissues. Histochemical analysis of adult brain from the two lines with highest lacZ mRNA levels showed neuron-specific, pan-neuronal beta-galactosidase activity. Developmental RNA and histochemical analyses showed parallel onset of transgene and endogenous NSE gene expression in various neuronal cell types, although the magnitude of NSE mRNA accumulation later in development was not matched by the transgene. These results suggest that cis-acting regulatory elements, subject to neuron-specific control, are located within 1.8 kb upstream from the NSE gene.

Animals↗

Long descending motor tract axons and their control of neck and axial muscles.

It has been tacitly assumed that a long descending motor tract axon consists of a private line connecting the cell of origin to a single muscle, as a motoneuron innervates a single muscle. However, this notion of a long descending motor tract referred to as a private line is no longer tenable, since recent studies have showed that axons of all major long descending motor tracts send their axon collaterals to multiple spinal segments, suggesting that they may exert simultaneous influences on different groups of spinal interneurons and motoneurons of multiple muscles. The long descending motor systems are divided into two groups, the medial and the lateral systems including interneurons and motoneurons. In this chapter, we focus mainly on the medial system (vestibulospinal, reticulospinal and tectospinal systems) in relation to movement control of the neck, describe the intraspinal morphologies of single long descending motor tract axons that are stained with intracellular injection of horseradish peroxidase, and provide evidence that single long motor-tract neurons are implicated in the neural implementation of functional synergies for head movements.

Afferent Pathways↗

Temporal and spatial coding of periodicity information in the inferior colliculus of awake chinchilla (Chinchilla laniger).

Amplitude modulation responses and onset latencies of multi-unit recordings and evoked potentials were investigated in the central nucleus of inferior colliculus (ICC) in the awake chinchilla. Nine hundred and one recording sites with best frequencies between 60 and 30 kHz showed either phasic (18%), tonic (25%), or phasic-tonic (57%) responses. Of 554 sites tested for responses to modulation frequencies 73% were responsive and 57% showed clear preference for a narrow range of modulation frequencies. Well defined bandpass characteristics were found for 32% of rate modulation transfer functions (rate-MTFs) and 36% of synchronization MTFs (sync-MTFs). The highest best modulation frequency (BMF) of a bandpass rate-MTF was 600 Hz. Neurons with phasic responses to best-frequency tones showed strong phase coupling to modulation frequencies and were dominated by bandpass rate-MTFs and sync-MTFs. Most neurons with tonic responses showed bandpass tuning only for sync-MTFs. Both BMFs and onset latencies changed systematically across frequency-band laminae of the ICC. Low BMFs and long latencies were located medially and high BMFs and short latencies laterally. Latency distributions obtained with evoked potentials to clicks showed a similar gradient to the multi-unit data. These findings are in line with previous findings in different animals including humans and support the hypothesis that temporal processing results in a topographic arrangement orthogonal to the spectral processing axis, thus forming a second neural axis of the auditory system.

Acoustic Stimulation↗

Expression of zebrafish fkd6 in neural crest-derived glia.

The zebrafish fkd6 gene is a marker for premigratory neural crest. In this study, we analyze later expression in putative glia of the peripheral nervous system. Prior to neural crest migration, fkd6 expression is downregulated in crest cells. Subsequently, expression appears initially in loose clusters of cells in positions corresponding to cranial ganglia. Double labelling with a neuronal marker shows that fkd6-expressing cells are not differentiated neurones and generally lie peripheral to neurones in ganglia. Later, expression appears associated with the posterior lateral line and other cranial nerves. For the posterior lateral line nerve, we show that fkd6-labeling extends caudally along this nerve in tight correlation with lateral line primordium migration and axon elongation. Expression in colourless mutant embryos is consistent with these cells being satellite glia and Schwann cells.

Animals↗

Inversion and transposition of Tc1 transposon of C. elegans in mammalian cells.

Tc1/mariner transposons are widespread in the eukaryotes. In vitro transposition test indicated that the transposase is the only protein that is needed in transpositions. It was shown later that the reconstructed Tc1-like transposon, "sleeping beauty" in fish, and the Tc1 transposon in C. elegans jumps in human cells. This discovery indicates that the Tc1/mariner transposon may be engineered as a somatic gene therapy vector if coupled with an efficient gene delivery system. We introduced the Tc1 transposon from C. elegans into different mammalian cell lines and detected the transposition events, indicating that Tc1 transposon functions in different mammalian cells. Interestingly, a high inversion frequency of the transposon was also detected, suggesting that this type of transposon may add variations to host genome when it is horizontally transferred into a new species.

Animals↗