Biomedical subjects
Publications and source records attributed to Andrew J King.
Mammals and birds appear to encode timing differences between the ears, a major cue for auditory localization, in fundamentally different ways. It now appears that results from different species can be accommodated within a single general framework.
We have used a slice preparation of the infant rat midbrain to examine converging inputs onto neurons in the deeper multisensory layers of the superior colliculus (dSC). Electrical stimulation of the superficial visual layers (sSC) and of the auditory nucleus of the brachium of the inferior colliculus (nBIC) evoked robust monosynaptic responses in dSC cells. Furthermore, the inputs from the sSC were found to be topographically organized as early as the second postnatal week and thus before opening of the eyes and ear canals. This precocious topography was found to be sculpted by GABAA-mediated inhibition of a more widespread set of connections. Tracer injections in the nBIC, both in coronal slices as well as in hemisected brains, confirmed a robust projection originating in the nBIC with distinct terminals in the proximity of the cell bodies of dSC neurons. Combined stimulation of the sSC and nBIC sites revealed that the presumptive visual and auditory inputs are summed linearly. Finally, whereas either input on its own could manifest a significant degree of paired-pulse facilitation, temporally offset stimulation of the two sites revealed no synaptic interactions, indicating again that the two inputs function independently. Taken together, these data provide the first detailed intracellular analysis of convergent sensory inputs onto dSC neurons and form the basis for further exploration of multisensory integration and developmental plasticity.
We have adapted a new approach for intrinsic optical imaging, in which images were acquired continuously while stimuli were delivered in a series of continually repeated sequences, to provide the first demonstration of the large-scale tonotopic organization of both primary and nonprimary areas of the ferret auditory cortex. Optical responses were collected during continuous stimulation by repeated sequences of sounds with varying frequency. The optical signal was averaged as a function of time during the sequence, to produce reflectance modulation functions (RMFs). We examined the stability and properties of the RMFs and show that their zero-crossing points provide the best temporal reference points for quantifying the relationship between the stimulus parameter values and optical responses. Sequences of different duration and direction of frequency change gave rise to comparable results, although in some cases discrepancies were observed, mostly between upward- and downward-frequency sequences. We demonstrated frequency maps, consistent with previous data, in primary auditory cortex and in the anterior auditory field, which were verified with electrophysiological recordings. In addition to these tonotopic gradients, we demonstrated at least 2 new acoustically responsive areas on the anterior and posterior ectosylvian gyri, which have not previously been described. Although responsive to pure tones, these areas exhibit less tonotopic order than the primary fields.
The organization of the cortical auditory system remains controversial. In particular, the extent to which there is regional specialization in the cortical processing of complex sound is unclear. Here, we ask whether we are currently asking the right questions of auditory cortex, or using the appropriate techniques to do so. A key factor that will promote such understanding in the future will be increasing dialogue between workers using electrophysiological recording methods to assess the response properties of single neurons and those using imaging techniques to map regional organization. In the future, further insights will be obtained by efforts to test hypotheses developed on the basis of one approach by the use of the other. Imaging can tell the neurophysiologists where to look, and work on single neurons can constrain network models based on imaging. There is a crucial need for better understanding of the anatomy of the auditory cortex in different species and for comparative studies that will underpin both approaches.
Lesion studies suggest that primary auditory cortex (A1) is required for accurate sound localization by carnivores and primates. In order to elucidate further its role in spatial hearing, we examined the behavioural consequences of reversibly inactivating ferret A1 over long periods, using Elvax implants releasing the GABA(A) receptor agonist muscimol. Sub-dural polymer placements were shown to deliver relatively constant levels of muscimol to underlying cortex for >5 months. The measured diffusion of muscimol beneath and around the implant was limited to 1 mm. Cortical silencing was assessed electrophysiologically in both auditory and visual cortices. This exhibited rapid onset and was reversed within a few hours of implant removal. Inactivation of cortical neurons extended to all layers for implants lasting up to 6 weeks and throughout at least layers I-IV for longer placements, whereas thalamic activity in layer IV appeared to be unaffected. Blockade of cortical neurons in the deeper layers was restricted to < or = 500 microm from the edge of the implant, but was usually more widespread in the superficial layers. In contrast, drug-free Elvax implants had little discernible effect on the responses of the underlying cortical neurons. Bilateral implants of muscimol-Elvax over A1 produced significant deficits in the localization of brief sounds in horizontal space and particularly a reduced ability to discriminate between anterior and posterior sound sources. The performance of these ferrets gradually improved over the period in which the Elvax was in place and attained that of control animals following its removal. Although similar in nature, these deficits were less pronounced than those caused by cortical lesions and suggest a specific role for A1 in resolving the spatial ambiguities inherent in auditory localization cues.
BACKGROUND: Polymorphonuclear leukocyte (PMNL) production of reactive oxygen species (ROS) has been linked to hemodialysis (HD) associated morbidity. The effect of dialyzer membrane type and reuse on PMNL function has not been clearly defined. METHODS: The present report is a cross-sectional study undertaken in a cohort of patients undergoing regular HD, at enrollment into the Hemodialysis (HEMO) Study, to study the association between patient and dialysis-related factors and PMNL function. PMNL function was assessed by measuring PMA- and N-formyl methionyl-leucyl-phenylalanine (fMLP) -induced respiratory burst, and phagocytic activity toward Staphylococcus aureus. RESULTS: PMNL from patients dialyzed with polysulphone (PS) or cuprophane (CU) membranes showed higher PMA-induced respiratory burst activity compared with those exposed to substituted cellulose (cellulose acetate, cellulose triacetate, CA/CT) membranes, regardless of dialyzer reuse. The use of bleach as a cleansing agent during reuse was associated with higher PMA-induced PMNL superoxide production, as was the use of renalin when compared to aldehydes. In a subgroup of patients using PS dialyzers, reuse itself was associated with higher fMLP-induced superoxide production. The type of bleach-germicide combination during reuse showed that use of renalin as a germicide was also associated with higher PMNL phagocytosis index. The number of years on HD correlated inversely with PMA-induced PMNL superoxide response. Weaker PMNL response to fMLP was associated with greater comorbidity and poor functional status as quantified by Index of Coexisting Diseases (ICED) and Karnofsky scores, respectively. CONCLUSION: Our results indicate that dialyzer membrane type and the reuse process influence the oxidative response of PMNL among HD patients. The implications of these observations on clinical morbidity need to be further evaluated in prospective studies.
BACKGROUND: Cytokine-orchestrated chronic inflammation plays a major role in long-term morbidity and mortality in patients with end-stage renal disease (ESRD) on hemodialysis (HD). In this cross-sectional study, we evaluated the association between specific alleles/genotypes and combinations of genotypes of interleukin (IL)-6, tumor necrosis factor-alpha (TNF-alpha), and IL-10 with indices of comorbidity, functional status, and other biological markers in a cohort of 183 ESRD patients recruited to the Hemodialysis (HEMO) Study from two Boston centers. METHODS: Genotyping was performed for single nucleotide polymorphisms in the promoter region of IL-6 (-174 G-->C), TNF-alpha (-308 G-->A), and IL-10 (-1082 G-->A). The relationship of specific genotypes to the index of coexistent disease (ICED) score (an index of comorbidity), Karnofsky Index (a measure of functional status), serum albumin, and nutritional indices (anthropometric measurements, body mass index, normalized protein catabolic ratio) were studied. Plasma IL-6 levels, as well as TNF-alpha and IL-10 production by endotoxin-stimulated peripheral blood mononuclear cells (PBMC), were also measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Patients with the high producer genotypes for the proinflammatory cytokines IL-6 (G/G and G/C) and TNF-alpha (G/A and A/A) had significantly higher comorbidity (ICED scores of > or =2) and lower functional scores (Karnofsky Index) compared with patients with the low producer genotypes for these cytokines (C/C and G/G, respectively). In contrast, patients with the high and intermediate producer genotypes (G/G and G/A) for the anti-inflammatory cytokine IL-10 had a higher Karnofsky Index compared with those with the low producer genotype (A/A). Serum albumin levels were lower in patients with the TNF-alpha high producer genotype (G/A and A/A) compared with those with the low producer genotype (G/A and A/A). On multivariate analysis, the IL-6 high producer genotypes were associated with an odds ratio (OR) of 4.87 for higher comorbidity (ICED scores > or =2) (P= 0.02), and 4.9 for lower Karnofsky Index (lower functional status) (P= 0.04) compared with patients with the low IL-6 producer genotypes. Similarly, the TNF-alpha high producer genotype was associated with increased odds for a higher ICED score, lower Karnofsky Index, and lower serum albumin compared with patients with the low producer genotype for this cytokine. In contrast, the IL-10 high/intermediate producer genotype was associated with increased odds for a higher Karnofsky Index (P= 0.05). Cytokine genotype combinations-the TNF-alpha high producer and IL-6 high producer genotype combination, and the IL-6 high producer and IL-10 low producer genotype combination-were independently associated with a higher ICED score. These genotype combinations, as well as the TNF-alpha high producer and IL-10 low producer genotype combination, were also associated with a lower Karnofsky Index. CONCLUSION: In ESRD patients on long-term HD, single nucleotide polymorphisms in the promoter region of the proinflammatory cytokines IL-6 and TNF-alpha, and the regulatory monokine IL-10, show a strong association with indices of comorbidity and function, and biological and nutritional markers.
The superior colliculus (SC) transforms both visual and nonvisual sensory signals into motor commands that control orienting behavior. Although the afferent and efferent connections of this midbrain nucleus have been well characterized, little is know about the intrinsic circuitry involved in sensorimotor integration. Transmission of visual signals from the superficial (sSC) to the deeper layers (dSC) of the SC has been implicated in both the triggering of orienting movements and the activity-dependent processes that align maps of different sensory modalities during development. However, evidence for the synaptic connectivity appropriate for these functions is lacking. In this study, we used a variety of anatomical and physiological methods to examine the functional organization of the sSC-dSC pathway in juvenile and adult ferrets. Axonal tracing in adult ferrets showed that, as in other species, sSC neurons project topographically to the dSC, providing a route for the transmission of visual signals to the multisensory output layers of the SC. We found that sSC axons terminate on dSC neurons that stain prominently for the NR1 subunit of the NMDA receptor, a subpopulation of which were identified as tectoreticulospinal projection neurons. We also show that the sSC-dSC pathway is topographically organized and mediated by monosynaptic excitatory synapses even before eye opening in young ferrets, suggesting that visual signals routed via the sSC may influence the activity of dSC neurons before the emergence of their multisensory response properties. These findings indicate that superficial- to deep-layer projections provide spatially ordered visual signals, both during development and into adulthood, directly to SC neurons that are involved in coordinating sensory inputs with motor outputs.
Terpenoids are important natural flavour compounds, which are introduced to beer via hopping. It has been shown recently that yeasts are able to biotransform some monoterpene alcohols. As a first step towards examining whether yeasts are capable of altering hop terpenoids during the brewing of beer, we investigated whether they were transformed when an ale and lager yeast were cultured in the presence of a commercially available syrup. Both yeasts transformed the monoterpene alcohols geraniol and linalool. The lager yeast also produced acetate esters of geraniol and citronellol. The major terpenoids of hop oil, however, were not biotransformed. Oxygenated terpenoids persisted much longer than the alkenes.
Auditory localization relies on the detection and interpretation of acoustic cues that change in value as the head and external ears grow. Here we show that the maturation of these structures is an important determinant for the development of spatial selectivity in the ferret auditory cortex. Spatial response fields (SRFs) of high-frequency cortical neurons recorded at postnatal days (P) 33-39 were broader, and transmitted less information about stimulus direction, than in older ferrets. They also exhibited slightly broader frequency tuning than neurons recorded in adult animals. However, when infant neurons were stimulated through virtual ears of adults, SRFs sharpened significantly and the amount of transmitted information increased. This improvement was predicted by a model that generates SRF shape from the localization cue values and the neurons' binaural spectrotemporal response properties. The maturation of spatial response characteristics in auditory cortex therefore seems to be limited by peripheral rather than by central factors.
Individual variations in head and outer ear size, as well as growth of these structures during development, can markedly alter the values of the binaural and monaural cues which form the basis for auditory localization. This study investigated individual differences in the directional component of the head-related transfer function of both adult and juvenile ferrets. In line with previous studies in humans and cats, intersubject spectral differences were found to be reduced by scaling one of the directional transfer functions on a log-frequency axis. The optimal scale factor correlated most highly with pinna cavity height. Optimal frequency scaling reduced interear spectral difference equally well for adult-juvenile comparisons as for comparisons between pairs of adult ears. This illustrates that the developmental changes in localization cue values should be at least partly predictable on the basis of the expected growth rate of the outer ear structures. Predictions of interaural time differences (ITDs) were also derived from the physical dimensions of the head. ITDs were found to be poorly fitted by the spherical head model, while much better predictions could be derived from a model based on von Mises spherical basis functions. Together, these findings show how more accurate estimates of spatial cue values can be made from knowledge of the dimensions of the head and outer ears, and may facilitate the generation of virtual acoustic space stimuli in the absence of acoustical measurements from individual subjects.
Recent studies in owls and ferrets seem to have identified the origin and nature of the visual signals that shape the development of the auditory space map in the midbrain, which ensures that the neural representations of both sensory modalities share the same topographic organization.
BACKGROUND: Studies have shown that survival factors including cytokines and growth factors delay apoptosis of human neutrophils via induction of the phosphatidylinositol-3 kinase (PI 3-K)/Akt pathway. In the present study, we explored whether complement fragment C5a has a modulatory effect on neutrophil apoptosis through this signaling pathway. METHODS: Human neutrophils were isolated and treated with C5a for up to 24 hours, with or without wortmannin, a PI 3-K inhibitor, and staurosporine, a caspase-9 activator. Apoptosis was quantified by flow cytometry, using propidium iodide nuclear staining, and confirmed by the detection of DNA fragmentation on gel electrophoresis. PI 3-K downstream signaling events were evaluated by measuring the expression of cytosolic total and phosphorylated Akt and Bad proteins by Western blot analyses, and caspase-9 activity. RESULTS: C5a inhibited neutrophil apoptosis in a dose- and time-dependent manner. The anti-apoptotic effects of C5a were markedly abrogated in the presence of wortmannin. Brief stimulation of neutrophils with C5a induced phosphorylation of Akt and Bad proteins through a PI 3-K-dependent pathway. Caspase-9 activity was minimal in C5a-treated cells, but markedly increased following PI 3-K inhibition by wortmannin. Finally, C5a reduced caspase-9 activity in staurosporine-treated cells. CONCLUSIONS: This study demonstrates that C5a inhibits neutrophil apoptosis via a PI 3-K signaling pathway. This effect may be an important mechanism that improves cell survival and function in the inflammatory milieu.
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The purpose of this study was to determine whether people could naturally produce urine sufficiently dilute to meet the federal criteria for a "substituted" specimen. The United States Department of Transportation Regulations (49 Code of Federal Regulations Part 40) defines a urine specimen as substituted if it has a creatinine concentration of < or = 5 mg/dL and a specific gravity of < or = 1.001 or > or = 1.020. These criteria have been criticized based on the contention that an insufficient number of specimens had been tested from the same urine sample for both creatinine and specific gravity measurements. We reviewed the results of 803,130 random urine specimens measured for creatinine and/or specific gravity in a hospital-based laboratory. In this database, 13,467 urine specimens had both creatinine and specific gravity measurements. None of these 13,467 paired urine specimens met the lower limit of specific gravity (< or = 1.001) and creatinine (< or = 5 mg/dL) criteria for a Department of Transportation substituted specimen. We also examined the medical records of those patients meeting even one of the two criteria; creatinine concentration < or = 5 mg/dL or specific gravity < or = 1.001. These patients were neonatal, moribund, or so severely ill that essentially none could have been among the working population. These data in patients with various pathologic states support our belief that normal individuals do not produce urine dilute enough to meet the lower limit of the specific gravity (< or = 1.001) and creatinine (< or = 5 mg/dL) required for meeting substituted specimen criteria. Eleven patients met the criteria for a substituted specimen, with elevated specific gravity of > or = 1.020 and creatinine concentration of < or = 5 mg/dL; however, these patients were seriously ill or terminally ill.
The current guidelines on dialysis adequacy in acute renal failure (ARF) are loosely defined and have been extrapolated from patients with end-stage renal disease. The objectives of this study were (1) to compare three methods of urea kinetic modeling measurement in patients with ARF receiving intermittent hemodialysis, (2) to compare prescribed to delivered dose of dialysis, and (3) to explore the factors that are associated with dialysis delivery. 'Single-pool' urea kinetic modeling was assessed by the Ureakin) software and the second-generation equation which uses a logarithmic estimate of spKt/V. 'Equilibrated' Kt/V (eKt/V) was calculated using the rate adjustment equation. The prescribed dose was derived using the manufacturer's specifications of the dialyzer clearance, prescribed time, actual delivered blood and dialysate flow, and estimates of volume of urea distribution. A total of 78 consecutive spKt/V measurements were obtained in 24 patients. The mean urea reduction ratio was 51 +/- 1%. The delivered spKt/V was significantly lower than that prescribed (0.87 +/- 0.03 or 0.83 +/- 0.03 vs. 1.28 +/- 0.05; p = 0.0001). The equilibrated Kt/V was markedly lower than the delivered spKt/V (0.73 +/- 0.03 vs. 0.83 +/- 0.03; p = 0.0001). Univariate analyses demonstrated that female gender, low body mass index, low predialysis weight, use of cellulose acetate dialyzers, and increased prescribed time were associated with increased odds of prescribed spKt/V > or =1.2. Similarly, old age, increased delivered time, and high cytokine production were associated with increased odds of delivered spKt/V > or =1.2. In summary, while the impact of delivered intermittent hemodialysis on the survival of patients with ARF remains to be determined, these results indicate that dialysis delivery is suboptimal in ARF, and empiric dosing should strongly consider factors related to lean body mass, including age and gender.
BACKGROUND: Interleukin-10 (IL-10), a potent regulatory monokine produced by activated mononuclear cells, provides an efficient autocrine mechanism for controlling proinflammatory cytokine synthesis. We hypothesized that defective synthesis of IL-10 could contribute to the inflammatory state in hemodialysis (HD) patients due to impaired feedback inhibition of proinflammatory cytokine production. METHODS: We compared peripheral blood mononuclear cell (PBMC) synthesis and transcription of IL-10 and TNF-alpha in 12 patients with end-stage renal disease on long-term maintenance HD and a control group of 10 healthy subjects. RESULTS: The synthesis of IL-10 by unstimulated PBMC was detectable in 5 of 12 (42%) HD patients as compared to 7 of 10 (70%) controls (p = 0.02). IL-10 synthesis in response to endotoxin (ET) by PBMC from HD patients was significantly lower when compared to the robust response in the control group (p = 0.008). Among the HD patients, there was a positive correlation between ET-stimulated IL-10 synthesis and the duration of time on dialysis. Unstimulated and ET-stimulated synthesis of TNF-alpha by PBMC did not differ between the 2 groups. In the HD patients, there was an inverse correlation between TNF-alpha and IL-10 synthesis by ET-stimulated PBMC, suggesting a regulatory effect of IL-10 on PBMC TNF-alpha synthesis. There was also an inverse correlation between plasma albumin and ET-stimulated TNF-alpha synthesis by PBMC among HD patients. TNF-alpha mRNA expression did not differ in HD patients relative to healthy controls. In contrast, when IL-10 mRNA from ET-stimulated PBMC was quantified, there was marked difference between the 2 groups indicating a transcriptional defect in IL-10 synthesis in PBMC from HD patients. CONCLUSION: Our observations indicate a marked abnormality in IL-10 synthesis by PBMC from HD patients probably related to a transcriptional defect. Low PBMC IL-10 synthesis may contribute to a chronic inflammatory state in these patients by defective feedback inhibition of proinflammatory cytokine production.