Sex differences in the influence of body mass index on anatomical architecture of brain networks.

BACKGROUND/OBJECTIVES: The brain has a central role in regulating ingestive behavior in obesity. Analogous to addiction behaviors, an imbalance in the processing of rewarding and salient stimuli results in maladaptive eating behaviors that override homeostatic needs. We performed network analysis based on graph theory to examine the association between body mass index (BMI) and network measures of integrity, information flow and global communication (centrality) in reward, salience and sensorimotor regions and to identify sex-related differences in these parameters. SUBJECTS/METHODS: Structural and diffusion tensor imaging were obtained in a sample of 124 individuals (61 males and 63 females). Graph theory was applied to calculate anatomical network properties (centrality) for regions of the reward, salience and sensorimotor networks. General linear models with linear contrasts were performed to test for BMI and sex-related differences in measures of centrality, while controlling for age. RESULTS: In both males and females, individuals with high BMI (obese and overweight) had greater anatomical centrality (greater connectivity) of reward (putamen) and salience (anterior insula) network regions. Sex differences were observed both in individuals with normal and elevated BMI. In individuals with high BMI, females compared to males showed greater centrality in reward (amygdala, hippocampus and nucleus accumbens) and salience (anterior mid-cingulate cortex) regions, while males compared to females had greater centrality in reward (putamen) and sensorimotor (posterior insula) regions. CONCLUSIONS: In individuals with increased BMI, reward, salience and sensorimotor network regions are susceptible to topological restructuring in a sex-related manner. These findings highlight the influence of these regions on integrative processing of food-related stimuli and increased ingestive behavior in obesity, or in the influence of hedonic ingestion on brain topological restructuring. The observed sex differences emphasize the importance of considering sex differences in obesity pathophysiology.

Altered viscerotopic cortical innervation in patients with irritable bowel syndrome.

BACKGROUND: Studies have demonstrated the existence of regional gray matter and white matter (WM) alterations in the brains of patients with irritable bowel syndrome (IBS), but the extent to which altered anatomical connectivity between brain regions is altered in IBS remains incompletely understood. METHODS: In this study, magnetic resonance imaging (MRI) and diffusion tensor imaging (DTI) were used to identify significant brain connectivity differences between IBS patients and healthy control (HC) subjects. Based on MRI and DTI volumes acquired from 66 IBS patients and 23 HC subjects, multivariate regression was used to investigate whether subject age, sex, cortical thickness, or the mean fractional anisotropy (FA) of WM connections innervating each location on the cortex could predict IBS diagnosis. KEY RESULTS: HC and IBS subjects were found to differ significantly within both left and right viscerotopic portions of the primary somatosensory cortex (S1), with the mean FA of WM bundles innervating S1 being the predictor variable responsible for these significant differences. CONCLUSIONS & INFERENCES: These preliminary findings illustrate how a chronic visceral pain syndrome and brain structure are related in the cohort examined, and because of their indication that IBS diagnosis is associated with anatomic neuropathology of potential neurological relevance in this patient sample.

Structural and connectomic neuroimaging for the personalized study of longitudinal alterations in cortical shape, thickness and connectivity after traumatic brain injury.

The integration of longitudinal brain structure analysis with neurointensive care strategies continues to be a substantial difficulty facing the traumatic brain injury (TBI) research community. For patient-tailored case analysis, it remains challenging to establish how lesion profile modulates longitudinal changes in cortical structure and connectivity, as well as how these changes lead to behavioral, cognitive and neural dysfunction. Additionally, despite the clinical potential of morphometric and connectomic studies, few analytic tools are available for their study in TBI. Here we review the state of the art in structural and connectomic neuroimaging for the study of TBI and illustrate a set of recently-developed, patient-tailored approaches for the study of TBI-related brain atrophy and alterations in morphometry as well as inter-regional connectivity. The ability of such techniques to quantify how injury modulates longitudinal changes in cortical shape, structure and circuitry is highlighted. Quantitative approaches such as these can be used to assess and monitor the clinical condition and evolution of TBI victims, and can have substantial translational impact, especially when used in conjunction with measures of neuropsychological function.

Major white matter fiber changes in medically intractable neocortical epilepsy in children: a diffusion tensor imaging study.

This study aimed to investigate the extent of microstructural changes in the major white matter fibers and to evaluate whether diffusion tensor imaging (DTI) adds any lateralizing information in children with medically intractable neocortical epilepsy secondary to focal cortical dysplasia. Patient group included twenty-three consecutively enrolled patients with medically intractable focal neocortical epilepsy and focal cortical dysplasia histopathologically confirmed. Thirteen patients (56.5%) had no visible lesion on the conventional magnetic resonance imaging (MRI). Fractional anisotropy (FA) was measured for regions of interest (ROIs) in each major white matter fiber. FA in patients was compared with eighteen age-matched healthy controls. Patient group had lower FA values at corpus callosum, bilateral inferior frontooccipital fasciculus (IFO), bilateral inferior longitudinal fasciculus (ILF) and left superior longitudinal fasciculus (SLF) compared to controls (p<0.05). In the left-side surgery group, the left SLF FA value was lower than controls, while in the right-side surgery group, the right SLF FA values were lower than controls (p<0.05). In the patient group as a whole, ipsilateral SLF FA was significantly lower than the contralateral SLF (p<0.05). Widespread decrease in FA values in the patients compared with the controls suggests that the pathologic changes extend diffusely to most major white matter tracts. In the patient group, the ipsilateral SLF to the seizure focus had greater change compared to the contralateral SLF. These data suggest that the detection of DTI abnormality has an added value to lateralization.

Estrogen increases smooth muscle expression of alpha2C-adrenoceptors and cold-induced constriction of cutaneous arteries.

Raynaud's phenomenon, which is characterized by intense cold-induced constriction of cutaneous arteries, is more common in women compared with men. Cold-induced constriction is mediated in part by enhanced activity of alpha(2C)-adrenoceptors (alpha(2C)-ARs) located on vascular smooth muscle cells (VSMs). Experiments were therefore performed to determine whether 17beta-estradiol regulates alpha(2C)-AR expression and function in cutaneous VSMs. 17beta-Estradiol (0.01-10 nmol/l) increased expression of the alpha(2C)-AR protein and the activity of the alpha(2C)-AR gene promoter in human cultured dermal VSMs, which was assessed following transient transfection of the cells with a promoter-reporter construct. The effect of 17beta-estradiol was associated with increased accumulation of cAMP and activation of the cAMP-responsive Rap2 GTP-binding protein. Transient transfection of VSMs with a dominant-negative mutant of Rap2 inhibited the 17beta-estradiol-induced activation of the alpha(2C)-AR gene promoter, whereas a constitutively active mutant of Rap2 increased alpha(2C)-AR promoter activity. The effects of 17beta-estradiol were inhibited by the estrogen receptor (ER) antagonist, ICI-182780 (1 micromol/l), and were mimicked by a cell-impermeable form of the hormone (estrogen:BSA) or by the selective ER-alpha receptor agonist 4,4',4'''-(4-propyl-[(1)H]-pyrazole-1,3,5-triyl)tris-phenol (PPT; 10 nmol/l) or the selective ER-beta receptor agonist 2,3-bis(4-hydroxyphenyl)-propionitrile (DPN; 10 nmol/l). Therefore, 17beta-estradiol increased expression of alpha(2C)-ARs by interacting with cell surface receptors to cause a cAMP/Rap2-dependent increase in alpha(2C)-AR transcription. In mouse tail arteries, 17beta-estradiol (10 nmol/l) increased alpha(2C)-AR expression and selectively increased the cold-induced amplification of alpha(2)-AR constriction, which is mediated by alpha(2C)-ARs. An estrogen-dependent increase in expression of cold-sensitive alpha(2C)-ARs may contribute to the increased activity of cold-induced vasoconstriction under estrogen-replete conditions.

Optimal testing parameters for blood cultures.

The effects of volume of blood, number of consecutive cultures, and incubation time on pathogen recovery were evaluated for 37,568 blood cultures tested with the automated BACTEC 9240 instrument (Becton Dickinson Diagnostic Instrument Systems) at a tertiary care center over the period of 12 June 1996 through 12 October 1997. When the results for this study were compared with previous data published for manual broth-based blood culture systems and patient samples obtained in the 1970s and 1980s, the following were found: (1) the percentage increase in pathogen recovery per milliliter of blood is less, (2) more consecutive blood culture sets over a 24-h period are required to detect bloodstream pathogens, and (3) a shorter duration of incubation is required to diagnose bloodstream infections. Guidelines developed in the 1970s and 1980s for processing and culturing blood may require revision.

Evolution of central respiratory chemoreception: a new twist on an old story.

Evolution of central respiratory chemosensitivity has been linked traditionally to the need for carbon dioxide regulation that accompanied the evolution of air breathing in terresterial animals. We examined the validity of this linkage by investigating the possibility of central chemoreception in air breathing fish that diverged from the amphibian lineage long before the appearance of terrestriality. We showed that the isolated brainstem preparation of the long nose gar (Lepisosteus osseus) produces a putative motor pattern for lung ventilation, which is responsive to CO(2). These findings, together with more inferential evidence, suggest an association between air breathing and central chemosensitivity in aquatic animals that spans the major branches in vertebrate phylogeny. Furthermore, developmental observations in tadpoles suggest that the neural substrates for central chemoreception exist in proximity to that for rhythm generation. We postulate that a primitive ancestral CPG, sensitive to CO(2) is conserved and is evidenced in the intrinsic coupling of respiratory CPG and central chemoreception in modern tetrapods.

Comparison of the BACTEC MYCO/F Lytic bottle to the isolator tube, BACTEC Plus Aerobic F/bottle, and BACTEC Anaerobic Lytic/10 bottle and comparison of the BACTEC Plus Aerobic F/bottle to the Isolator tube for recovery of bacteria, mycobacteria, and fungi from blood.

The BACTEC MYCO/F Lytic blood culture bottle (Becton Dickinson Diagnostic Instrument Systems, Sparks, Md.) is designed to optimize the recovery of fungi and mycobacteria; however, this bottle also supports the growth of most aerobic bacteria. We compared the MYCO/F Lytic bottle with two other BACTEC bottles and the Isolator system for the recovery of bacteria as well as fungi and mycobacteria from blood. A total of 6,108 blood culture sets were inoculated with blood obtained from adult patients. Twenty-five to 28 ml of blood collected by a phlebotomy team for each blood culture set was randomly distributed into each of four blood culture receptacles: the Isolator tube (Wampole Laboratories, Cranbury, N.J.) and three BACTEC bottles: the MYCO/F Lytic bottle, the BACTEC Plus Aerobic/F bottle, and the BACTEC Anaerobic Lytic/10 bottle. The sediment from the Isolator tube was inoculated onto chocolate agar (CA), brain heart infusion agar (BHI), and Sabouraud dextrose agar (SDA) and into a BACTEC 13A bottle. Incubation durations were as follows: MYCO/F Lytic bottle, 42 days; Plus Aerobic/F bottle, 5 days; Anaerobic Lytic/10 bottle, 5 days; sediment from Isolator tube on CA, 3 days; sediment from Isolator tube on BHI, 30 days; sediment from Isolator tube on SDA, 30 days; and sediment from Isolator tube in a BACTEC 13A bottle, 42 days. Two isolates of Histoplasma capsulatum were recovered from the Isolator tube only. Three isolates of Mycobacterium tuberculosis complex were recovered: two isolates from the MYCO/F Lytic bottle only and one isolate from the Isolator tube (whose sediment was inoculated into the BACTEC 13A bottle) only. Two isolates of Cryptococcus neoformans were recovered: one from the MYCO/F Lytic bottle only and the other from the MYCO/F Lytic bottle and the Isolator tube (whose sediment was inoculated into the BACTEC 13A bottle). For potential pathogens overall, there was a statistical difference in recovery that favored the Isolator system over the MYCO/F Lytic bottle (P = 0.0015), including statistically significant differences for Staphylococcus aureus (P = 0.0001) and Streptococcus pneumoniae (P = 0.0313). However, there was no statistically significant difference between the two blood culture systems when detection of bloodstream infection was considered. The time to detection for all potential pathogens combined was less for the MYCO/F Lytic bottle than for the Isolator system (P = 0.0004). Overall, the potential pathogen recovery was greater for the BACTEC Plus Aerobic/F bottle than for either the Isolator system (P = 0.0003) or the MYCO/F Lytic bottle (P = 0.0001). However, the BACTEC Plus Aerobic/F bottle did not recover M. tuberculosis, H. capsulatum, or C. neoformans isolates. The combination of the Isolator system and MYCO/F Lytic bottle may be useful as a selective blood culture method to optimize the recovery of fungi and mycobacteria from blood. Compared with the manual Isolator system, the MYCO/F Lytic system has the advantage of less preanalytic processing and continuous automated monitoring of bottles for growth by the BACTEC 9240 instrument.

Direct identification of bacteria from positive blood cultures by amplification and sequencing of the 16S rRNA gene: evaluation of BACTEC 9240 instrument true-positive and false-positive results.

In a previous study which evaluated the BACTEC 9240 automated blood culture system (Becton Dickinson Diagnostic Instrument Systems, Sparks, Md.), we noted a 1.3% "instrument false-positive" rate. That is, the BACTEC system signaled that a bottle (BACTEC Plus Aerobic/F bottle or BACTEC Anaerobic Lytic/10 bottle) culture was positive but a Gram stain was negative and there was no growth of bacteria or yeasts on subculture to chocolate agar. Furthermore, from the same sample of blood, cultures for fungi using the Isolator blood culture system (Wampole Laboratories, Cranbury, N.J.) were negative for growth. For the present study, we evaluated 76 instrument false-positive samples for the presence of 16S ribosomal DNA using the MicroSeq 500 kit (PE Biosystems, Foster City, Calif.). These samples also were negative for fungi by the Isolator method. This kit has a PCR module and sequencing module for the amplification and sequencing of the 16S RNA gene and provides a database for sequence alignment and identification of bacteria. To optimize the assay, we evaluated the effect of adding 0.5% bovine serum albumin to the sample from blood culture bottles and found that it decreased the effects of inhibitors on the PCR. Two control groups of blood culture specimens were also evaluated. One group (n = 45) were "instrument true positives"; the instrument signaled positive, and subsequent Gram stains were positive and subcultures on chocolate agar grew bacteria. The other group (n = 20) were "instrument true negatives"; the instrument signaled negative, the Gram stain was negative, and subcultures on chocolate agar and from the Isolator tube on fungal media showed no growth. None of the 76 instrument false-positive samples had evidence for 16S rRNA gene sequences. All of the instrument true-positive samples and all of the instrument true-negative specimens were positive and negative, respectively, using the MicroSeq 500 kit. Total peripheral white blood cell counts were statistically significantly higher for patients who had instrument false-positive results than for patients who had instrument true-positive or true-negative results (P = 0.001). We conclude that instrument false positives signaled by the BACTEC 9240 system are not due to bacteria in the blood culture samples.

Sites of respiratory rhythmogenesis during development in the tadpole.

During ontogeny, amphibian larvae experience a dramatic alteration in the motor act of breathing as the premetamorphic gill breather develops into the postmetamorphic lung ventilator. We tested the hypothesis that the site of lung rhythmogenesis relocates during metamorphosis by recording fictive lung ventilation before and after transecting the in vitro brain stem of pre- and postmetamorphic Rana catesbeiana into four segments. In premetamorphic tadpoles, the two caudalmost brain stem segments combined proved to be the minimum brain stem configuration necessary and sufficient for lung burst generation. In the postmetamorphic counterpart, this function was supplied by the combination of the two rostralmost brain stem segments. In the postmetamorphic brain stem, a 500-microm segment lying just rostral to cranial nerve IX conveys rhythmogenic capability to neighboring rostral or caudal segments. We conclude that lung rhythmogenic capability translocates rostrally during development as the tadpole shifts from gill to lung ventilation.

Location of central respiratory chemoreceptors in the developing tadpole.

The location of central respiratory chemoreceptors in amphibian larvae may change as the central chemoreceptive function shifts from driving gill to driving lung ventilation during metamorphosis. We examined this possibility in the in vitro brain stem of the pre- and postmetamorphic Rana catesbeiana tadpole by microinjecting hypercapnic artificial cerebrospinal fluid (aCSF) while recording fictive lung ventilation. The rostral and caudal brain stem were separately explored systematically using injections of 11 nl of aCSF equilibrated with 100% CO2 that transiently acidified a 500-microm region, producing a maximum reduction in pH of 0.23 +/- 0.06 at the site of injection. In postmetamorphic tadpoles, chemoreceptive sites were concentrated in the rostral compared with the caudal brain stem. No such segregation was observed in the premetamorphic tadpole. We conclude that, as in lung rhythmogenic function, respiratory chemosensitivity emerges rostrally in the amphibian brain stem during development.

Cyclic cystography: diagnostic yield in selected pediatric populations.

PURPOSE: To test the hypothesis that the diagnostic yield of cyclic cystography is related to the prevalence of vesicoureteral reflux (VUR) in the population being evaluated. MATERIALS AND METHODS: Two groups of children were examined prospectively: 124 with severe urinary tract infection, defined as patient hospitalization or a maximum temperature greater than 39.5 degrees C, and 135 with previously diagnosed VUR. Nuclear cystography was performed in 249 patients, and fluoroscopic cystography was performed in 10. If VUR was not seen during the first cycle of bladder filling and voiding, a second cycle was performed. RESULTS: VUR was present during cycle 1 in 40 (32%) of 124 patients with severe urinary tract infection and 90 (67%) of 135 children in the VUR follow-up group (P < .001). VUR was demonstrated during cycle 2 in seven (9%) of 76 of the severe urinary tract infection group and eight (24%) of 34 of the VUR follow-up group (P = .045). Of 15 patients with VUR during cycle 2, two had grade III VUR and 13 had grade I or II VUR. CONCLUSION: The second cycle of cyclic cystography has a higher diagnostic yield in patients undergoing VUR follow-up than in patients with severe urinary tract infection. The decision to perform a second cycle of bladder filling and voiding should take into account the pretest probability of VUR in the child being examined.

The fictively breathing tadpole brainstem preparation as a model for the development of respiratory pattern generation and central chemoreception.

Spontaneous high-frequency, low-amplitude and low-frequency, high-amplitude efferent bursting patterns of cranial and spinal motor nerve activity in the in vitro brainstem preparation of the bullfrog tadpole Rana catesbeiana have been characterized as fictive gill and lung ventilation, respectively (Gdovin MJ, Torgerson CS, Remmers JE). Characterization of gill and lung ventilatory activity in cranial nerves in the spontaneously breathing tadpole Rana catesbeiana, FASEB J 1996;10(3):A642; Gdovin MJ, Torgerson CS, Remmers JE. Neurorespiratory pattern of gill and lung ventilation in the decerebrate spontaneously breathing tadpole, Respir Physiol 1998;113:135 146; Pack AI, Galante RJ, Walker RE, Kubin LK, Fishman AP. Comparative approach to neural control of respiration, In: Speck DF, Dekin MS, Revelette WR, Frazier DT, editors. Respiratory Control Central and Peripheral Mechanisms. Lexington: University of Kentucky Press, 1993:52-57). In addition, the ontogenetic dependence of central respiratory chemoreceptor stimulation on fictive gill and lung ventilation has been previously described (Torgerson CS, Gdovin MJ, Remmers JE. Fictive gill and lung ventilation in the pre- and post-metamorphic tadpole brainstem, J Neurophysiol 1998, in press). To investigate the neural substrates responsible for central respiratory rhythm generation of gill and lung ventilation in the developing tadpole, we recorded efferent activities of cranial nerve (CN) V, VII, and X and spinal nerve (SN) II during changes in superfusate PCO2 before and after multiple transection of the in vitro brainstem. The brainstem was transected between CN VIII and IX and the response to changes in PCO2 was recorded. A second transection was then made between the caudal margin of CN X and rostral to SN II. Preliminary data reveal that robust gill ventilation was recorded consistently only if the segment of brainstem included CN X, whereas the loci capable of eliciting fictive lung bursting patterns appeared to differ depending on developmental stage. These data demonstrate that the neural substrate required for fictive gill and lung ventilation exists in anatomically separate regions such that the gill central pattern generator (CPG) is located in the caudal medulla at the level of CN X throughout development, whereas the location of the lung CPG is located more rostrally at the level of CN VII in the post-metamorphic larva. Both in vivo and in vitro studies revealed two distinct neural bursting patterns associated with gill and lung ventilation. Sequential activation of CN V, VII, X were observed during gill ventilation of in vivo and fictive gill ventilation in vitro, whereas these nerve activities, along with SN II displayed more synchronous bursting patterns of activation during lung ventilation and fictive lung breaths.

Neurorespiratory pattern of gill and lung ventilation in the decerebrate spontaneously breathing tadpole.

A decerebrate, spontaneously breathing tadpole preparation (Taylor-Kollros stages 16-19) was used to test the general hypothesis that the efferent bursting activities of cranial nerves (CN) V, VII and spinal nerve (SN) II are respiratory in nature, and, in particular, to identify separate and specific neural correlates of gill and lung ventilation. Oropharyngeal pressure (POP), intrapulmonary pressure (PIP), electromyogram (EMG) of the buccal levator muscle (interhyoideus), and efferent neural activities of CN V, CN VII and SN II were recorded while the animal was exposed to hyperoxia (100% inspired O2), normoxia (21% inspired O2), and hypoxia (10, 5 and 0% inspired O2). Gill ventilation, indicated by fluctuations in POP at constant PIP, was characterized by high-frequency, low-amplitude bursts of action potentials in CN V and VII and interhyoideus EMG without phasic activity in SN II. Lung breaths, indicated by oscillations in POP and PIP were characterized by large bursts in EMG, CN V and VII together with a large burst in SN II. The amplitude of the moving average of nerve activities associated with lung ventilation was significantly larger than those associated with gill ventilation. During gill ventilation, the burst in CN V led that in CN VII, and both preceded the rise in POP. By contrast, a more synchronous neural burst onset pattern was observed during lung ventilation. The results document the neural, muscular, and mechanical characteristics of gill and lung ventilation in the tadpole, and establish bursting activity in SN II as a specific marker for lung ventilation in the metamorphic tadpole.

Fictive gill and lung ventilation in the pre- and postmetamorphic tadpole brain stem.

The pattern of efferent neural activity recorded from the isolated brain stem preparation of the tadpole Rana catesbeiana was examined to characterize fictive gill and lung ventilations during ontogeny. In vitro recordings from cranial nerve (CN) roots V, VII, and X and spinal nerve (SN) root II of premetamorphic tadpoles showed a coordinated sequence of rhythmic bursts occurring in one of two patterns, pattern1, high-frequency, low-amplitude bursts lacking corresponding activity in SN II and pattern 2, low-frequency, high-amplitude bursts with coincident bursts in SN II. These two patterns corresponded to gill and lung ventilatory burst patterns, respectively, recorded from nerve roots of decerebrate, spontaneously breathing tadpoles. Similar patterns were observed in brain stem preparations from postmetamorphic tadpoles except that they showed a greater frequency of lung bursts and they expressed fictive gill ventilation in SN II. The laryngeal branch of the vagus (Xl) displayed efferent bursts in phase with gill and lung activity, suggesting fictive glottal constriction during gill ventilation and glottal dilation during lung ventilation. The fictive gill ventilatory cycle of pre- and postmetamorphic tadpoles was characterized by a rostral to caudal sequence of CN bursts. The fictive lung ventilatory pattern in the premetamorphic animal was initiated by augmenting CN VII discharge followed by synchronous bursts in CN V, X, SN II, and Xl. By contrast, postmetamorphic patterns of fictive lung ventilation were characterized by lung burst activity in SN II that preceded burst onset in CN V and followed the lead burst in CN VII. We conclude that recruitment and timing of pattern 1 and pattern 2 rhythmic bursts recorded in vitro closely resemble that recorded during spontaneous respiratory behavior, indicating that the two patterns are the neural equivalent of gill and lung ventilation, respectively. Further, fictive gill and lung ventilatory patterns in postmetamorphic tadpoles differ in burst onset latency from premetamorphic tadpole patterns and resemble fictive oropharyngeal and pulmonary burst cycles in adult frogs.

Clinical comparison of BACTEC 9240 plus aerobic/F resin bottles and the isolator aerobic culture system for detection of bloodstream infections.

The Plus Aerobic/F resin bottle of the BACTEC 9240 automated blood culture system (Becton Dickinson Diagnostic Instrument Systems, Sparks, Md.) was compared with aerobic culture of the Isolator system (Wampole Laboratories, Cranbury, N.J.) for the detection of bloodstream microorganisms from 6,145 blood cultures collected from adult patients with suspected septicemia. The BACTEC resin bottles were incubated for 7 days, and the sediment from the Isolator tube was inoculated to sheep blood and chocolate agars which were incubated for 72 h and to inhibitory mold, brain heart infusion, and Sabouraud agars which were incubated for 21 days. A total of 622 microorganisms were recovered from 583 blood cultures. The BACTEC resin bottle recovered statistically significantly more pathogens overall than the Isolator system (P = 0.0006). When individual pathogens isolated from either system for a 7-day study period were assessed, it was determined that the BACTEC resin bottle detected statistically significantly more isolates of Staphylococcus aureus (P = 0.0113) and coagulase-negative Staphylococcus spp. (P = 0.0029) than the Isolator system. The BACTEC resin bottle also detected statistically significantly more bloodstream infections (septic episodes) caused by coagulase-negative Staphylococcus spp. (P = 0.0146). The Isolator system recovered statistically significantly more contaminants overall (P < 0.0001), and among this group of microorganisms, recovered statistically significantly more Bacillus spp. (P < 0.0001), coagulase-negative Staphylococcus spp. (P < 0.0001), and viridans group Streptococcus spp. (P = 0.0156). The Isolator system detected statistically significantly more isolates of Histoplasma capsulatum (P = 0.004), but all of these isolates were detected at > or = 7 days of incubation of fungal plates, i.e., after the system to system comparison study period (7 days). In blood culture sets which produced growth of the same pathogen in both systems, there was a statistically significant difference in median time to detection for all pathogens combined favoring the BACTEC resin bottle over the Isolator tube (P < 0.05). When assessing individual microorganisms, the median times for detection of S. aureus, Enterococcus spp., and Pseudomonas spp. were all statistically significantly less for the BACTEC system (P < 0.05). The BACTEC instrument had 79 (1.3%) false positive signals. The BACTEC system required less processing time than the Isolator system and eliminates the hands-on time for detection of positive cultures required with the Isolator system.

Depth profiles of pH and PO2 in the in vitro brainstem preparation of the tadpole Rana catesbeiana.

Extracellular pH and PO2 was recorded in the isolated in vitro brainstem of the metamorphic tadpole, Rana catesbeiana while the brainstem preparation was superfused with oxygenated mock cerebrospinal fluid of pH = 7.8, PCO2 = 17 Torr, PO2 = 600 Torr at 23 degrees C. Using pH and PO2 microelectrodes, the ventral medullary surface was penetrated at midline and lateral sites between cranial nerves V and X. Mean pH and PO2 gradients of 0.07 pH units/100 microns and 60 Torr/100 microns were detected in the superfusate, 100-200 microns above the ventral surface of the brainstem. These gradients remained virtually constant for the first 100-200 microns below the medullary surface. Beyond this level, pH and PO2 gradients decreased in a curvilinear fashion. For midline tracts, minimum values of pH and PO2 (7.58 +/- 0.05 and 323 +/- 31 Torr) were reached at a depth of 500-750 microns, whereas for lateral tracts, mean minimum values of pH and PO2 (7.34 +/- 0.12 and 240 +/- 68 Torr), were recorded at 850-900 microns. With further electrode advancement, pH and PO2 gradients in both midline and lateral tracts reversed as levels began to increase. Between CN V and X, lateral width was 4.34 +/- 0.57 mm, while dorsal-ventral thickness in midline and lateral regions was 0.92 +/- 0.21 and 1.31 +/- 0.22 mm, respectively. Overall, the in vitro tadpole brainstem provides a robust neural preparation which, although moderately acidic, is well oxygenated throughout all tissue layers.

Ontogeny of central chemoreception during fictive gill and lung ventilation in an in vitro brainstem preparation of Rana catesbeiana

An isolated brainstem preparation of the bullfrog tadpole, Rana catesbeiana, displays coordinated rhythmic bursting activities in cranial nerves V, VII and X in vitro. In decerebrate, spontaneously breathing tadpoles, we have previously shown that these bursts correspond to fluctuations in buccal and lung pressures and to bursts of activity in the buccal levator muscle H3a. This demonstrates that the rhythmic bursting activities recorded in vitro represent fictive gill and lung ventilation. To investigate the ontogeny of central respiratory chemoreception during the transition from gill to lung ventilation, we superfused the isolated brainstems of four larval stage groups with oxygenated artificial cerebrospinal fluid at various levels of PCO2. We measured shifts in the pattern of fictive respiratory output and the response to central hypercapnic stimulation throughout development. At normal PCO2 (2.3 kPa), stage 3­9 tadpoles displayed rhythmic neural bursts associated with gill ventilation, while stages 10­14 and 15­19 tadpoles produced oscillating bursting activity associated with both gill and lung respiration, and tadpoles at stages 20­25 displayed neural activity predominantly associated with lung ventilation. In stage 3­9 tadpoles, variations in PCO2 of the superfusate (0.5­6.0 kPa) caused almost no change in fictive gill or lung ventilation. By contrast, stage 10­14 tadpoles showed a significant hypercapnic response (P<0.05) in the amplitude and frequency of fictive gill ventilation, which was accompanied by a significant increase (P<0.05) in the burst amplitude and respiratory output of cranial nerve X over that occurring at all other stages. The amplitude and frequency of fictive gill ventilation in stages 15­19 increased significantly (P<0.05) in response to pH reduction, but became insensitive to hypercapnia at stages 20­25. The frequency of fictive lung ventilation was unresponsive to hypercapnia in stage 10­14, increased significantly by stage 15­19 (P<0.05) and became maximal (P<0.05) in stages 20­25. Overall, we describe the ontological development of central respiratory chemoreceptors driving respiratory output in the larval amphibian, demonstrating transfer in central chemoreceptive influence from gill to lung regulation during metamorphic stages. In addition, we provide novel evidence for the stimulatory influence of central chemoreceptors on fictive gill ventilation in response to CO2.

Clinical comparison of difco ESP, Wampole isolator, and Becton Dickinson Septi-Chek aerobic blood culturing systems.

The ESP 80A aerobic blood culture of the ESP automated blood culture system (Difco Laboratories. Detroit, Mich.) was compared with two manual aerobic blood culture systems, the Isolator (Wampole Laboratories, Cranbury, N.J.) and the Septi-Chek (Becton Dickinson, Cockeysville, Md.) systems, for the detection of bloodstream microorganisms from 5,845 blood samples for culture collected from adult patients with suspected septicemia. The bottles were incubated for 7 days, and the sediment from the Isolator tube was inoculated onto solid medium and this medium was incubated for 72 h. A total of 609 microorganisms were recovered from 546 blood cultures. There was no statistically significant difference in the total recovery of microorganisms for the ESP 80A system when compared with that for the Septi-Chek system (P = 0.083); however, the Isolator system recovered significantly more microorganisms overall than either the ESP 80A (P < 0.001) or the Septi-Chek (P < 0.001) system. When assessing individual probable pathogens, the Isolator system detected statistically significantly more Staphylococcus aureus and Candida spp. than either the ESP 80A or the Septi-Chek system (P < 0.05). Similarly, the Isolator system detected statistically significantly more bloodstream infections (septic episodes) caused by S. aureus and Candida spp. than either the ESP 80A or the Septi-Chek system (P < 0.05). In blood culture sets which produced growth of the same probable pathogens in the ESP 80A and the Isolator systems, there was no statistically significant difference in the median times to detection for all pathogens combined (P = 0.067). However, a similar comparison showed the Isolator and the ESP 80A systems to have statistically significantly shorter median detection times for all pathogens combined (P < 0.001) when they were independently compared with the Septi-Chek system. The ESP 80A system had 29 (0.5%) false-positive signals. The ESP system required less processing time than the Isolator system and eliminates the hands-on time for the detection of positive cultures required by the manual systems.