A comparison between continuous central venous pressure measurement from right atrium and abdominal vena cava or common iliac vein.

OBJECTIVE: To determine the accuracy of mean continuous central venous pressure (CVP) measurements in the abdominal vena cava. DESIGN: We simultaneously measured the CVP at the superior vena cava or right atrium and at the abdominal vena cava or common iliac vein. The study was conducted at the pediatric intensive care unit of a major university-affiliated medical center. PATIENTS: Nine patients, aged 6 months to 14 years, were included in our study. MEASUREMENTS AND RESULTS: Eleven continuous recordings of 12 to 68 min were taken, eight of them while the children were mechanically ventilated. Mean overall CVP ranged from 3 to 30 mmHg. A total of 519 simultaneous recordings were made, of which 515 (99.2%) were within the accepted limits of agreement of +/- 2 mmHg: 301 (58%) with delta CVP of +/- 0 mmHg, 189 (36,4%) with delta CVP of +/- 1 mmHg, and 25 (4.8%) with delta CVP of +/- 2 mmHg. The mean pressure difference was -0.22 +/- 1.52 mmHg. Accuracy was maintained within all ranges of CVP (3-10, 11-20, and 21-30 mmHg) and was not influenced by mechanical ventilation or abdominal fluid collection. CONCLUSION: In children with no obstruction of blood flow from the abdominal vena cava to the right atrium, the pressure in the abdominal vena cava or common iliac vein accurately reflects the pressure in the right atrium.

SgfI, a new type-II restriction endonuclease that recognizes the octanucleotide sequence 5'-GCGAT/CGC-3'.

A new restriction endonuclease (ENase), SgfI, has been isolated from the bacterium Streptomyces sp. SgfI recognizes the 8-bp palindrome 5'-GCGATCGC-3' and cleaves double-stranded DNA after the T in this sequence, producing a two-base 3' overhang compatible with PvuI termini. SgfI is a rare-cutting ENase and should be useful for megabase mapping experiments.

The spatial organization of the peripheral olfactory system of the hamster. Part I: Receptor neuron projections to the main olfactory bulb.

The spatial organization of projections from olfactory receptor neurons to the main olfactory bulb (MOB) was studied in hamsters by using fluorescent stilbene isothiocyanates as retrograde tracers. Injections confined to small sectors of the MOB produce labeling of receptor neurons that is more restricted circumferentially (i.e., with respect to the medial-lateral and dorsal-ventral axes) than longitudinally (i.e., with respect to the rostral-caudal axis) along the mucosal sheet. This restricted labeling is also discontinuous, giving an initial impression that the peripheral input is only crudely organized with respect to the medial-lateral and dorsal-ventral axes of the nasal cavity. However, from analyses of serial sections, it is apparent that each set of mucosal segments shares convergent projections to a circumferential quadrant of the MOB with other segments that are positioned around a common domain of the nasal cavity airspace. The primary afferent projections to the MOB, thus, are organized rhinotopically (i.e., with respect to the three-dimensional position of receptor neurons in olfactory space) rather than mucosotopically.

The spatial organization of the peripheral olfactory system of the hamster. Part II: Receptor surfaces and odorant passageways within the nasal cavity.

The spatial organization of olfactory receptor surfaces and odorant passageways within the nasal cavity was studied in hamsters through descriptive and morphometric analyses of a complete stereotaxically defined series of coronal, sagittal, and horizontal sections through the snout. These analyses reveal that the caudal two-thirds of each cavity is divided into two longitudinally oriented medial and lateral channels. The olfactory mucosa that lines these two channels projects selectively onto the medial and lateral halves of the main olfactory bulb (MOB), respectively. Moreover, the ethmoturbinates of the caudal recesses create highly convoluted channels, lined by ventrally projecting mucosa, that lie ventral, lateral, and dorsal to a relatively smooth central channel lined by dorsally projecting mucosa. The rhinotopic map makes equivalent representations of medial and lateral olfactory space to the MOB but gives the smooth space lined by dorsally projecting mucosa a disproportionately larger representation on the MOB than the convoluted space lined by the more expansive ventrally projecting mucosa. Recent descriptions of the spatial distribution of probes for odorant receptor proteins conform closely to this organization, giving credence to the idea that rhinotopy is a basis for representing to the MOB the specific molecular features of odorant molecules.

Bca77I: a new type-II restriction endonuclease from Bacillus caldolyticus cutting at the (A/T) increases CCGG(A/T) site.

We describe a new restriction enzyme recognizing a degenerate hexanucleotide sequence and cleaving the 5'-W increases CCGGW site (W = A or T). This enzyme cuts with high efficiency all four permutations of this sequence; ACCGGA (TCCGGT on the opposite strand), ACCGGT and TCCGGA. Methylation of the external cytosine completely blocks restriction while methylation of the internal cytosine only decreases the rate of restriction activity.

Bst71I: an isoschizomer of the type-IIS restriction enzyme, BbvI, recognizing the GCAGC(8/12) site.

A new type-IIS restriction enzyme, Bst71I, with the specificity 5'-GCAGC(N)8/3'-CGTCG(N)12 was isolated from Bacillus stearothermophilus (Promega No. 71). This enzyme is an isoschizomer of BbvI with somewhat improved characteristics for use by molecular biologists.

Dactinomycin potentiation of radiation pneumonitis: a forgotten interaction.

No mention of dactinomycin potentiation of pulmonary radiation was found in a review of the literature of the past 12 years. Before that, this complication was well described and investigators had calculated that dactinomycin increased the toxic effect of lung radiation by a factor of 1.3 and reduced the radiation tolerance of the lung by at least 20%. An example of such a toxic effect is described in the treatment of a 7-year-old girl with lung metastases from Ewing's sarcoma. The chemotherapy protocol followed contained cyclophosphamide, vincristine, dactinomycin, adriamycin, cisplatinum, VP16, and radiotherapy. The treatment was associated with fatal pulmonary fibrosis following the reintroduction of dactinomycin after radiotherapy. Our experience suggests that there is clinical significance to this complication in sarcoma therapy when dactinomycin-containing protocols are used with radiation in the treatment of pulmonary metastases.

Multiple proteins are produced from the dec-1 eggshell gene in Drosophila by alternative RNA splicing and proteolytic cleavage events.

The defective chorion-1 gene (dec-1) in Drosophila encodes follicle cell proteins necessary for proper eggshell assembly. A distinctive feature of the gene is the production of multiple products by both alternative RNA splicing and proteolytic processing events. DNA and protein sequencing studies have revealed several dec-1 protein products. The predominant translation product, fc106, has a vitelline membrane-like N-terminal domain followed by a glutamine, methionine-rich central region, largely in the form of 26 amino acid repeats. During late stage 10 the N-terminal portion of fc106 is cleaved, yielding s80, a major eggshell protein. Conceptual translation of the DNA sequence as well as molecular analyses of several dec-1 mutants suggest that the less abundant alternatively spliced RNAs encode primary translation products with different carboxy terminal ends. These results are discussed with respect to previous genetic analyses of dec-1 mutants as well as with respect to potential protein-protein interactions which may underlie stabilization of this complex extracellular structure.

MAP 1A and MAP 1B are structurally related microtubule associated proteins with distinct developmental patterns in the CNS.

Five high-molecular-weight microtubule-associated proteins (MAPs) were identified in brain tissue in previous work from this laboratory (Bloom et al., 1984). These proteins were termed MAP 1A, 1B, 1C, 2A, and 2B. The MAP 1's differed from the MAP 2's, and showed little evidence of interrelationship on the basis of immunological and biochemical comparison. We report here that MAP 1A and MAP 1B are, in fact, related at the level of subunit composition. Immunoprecipitation of the individual MAPs showed that both contained low-molecular-weight subunits of Mr 30,000 and Mr 19,000 (light chains 1 and 3). An additional subunit, light chain 2 (Mr 28,000), was primarily found in preparations of MAP 1A. The light chains co-sedimented with microtubules after chymotryptic digestion of the MAPs. This suggested an association of the light chains with the microtubule binding domains of the MAPs, which are identified here as distinct fragments of Mr 60,000 for MAP 1A and 120,000 for MAP 1B. A panel of monoclonal anti-MAP 1A and anti-MAP 1B antibodies, including one that reacts with a common phosphorylated epitope, was used to examine the distribution of these proteins in the developing rat brain and spinal cord. MAP 1B was found to be abundant in the newborn brain and to decrease with development, in contrast to MAP 1A which increased with development. By immunohistochemistry MAP 1B was found to be highly concentrated in developing axonal processes in the cerebellar molecular layer, the corticospinal tract, the mossy fibers in the hippocampus, and the olfactory nerve. Of particular interest, the mossy fiber and olfactory nerve staining persisted in the adult, indicating continued outgrowth of the mossy fibers as well as olfactory nerve axons. MAP 1A staining was, in contrast, weak or absent in developing axonal fibers but moderate in mature axons and intense in developing and mature dendritic processes. Our results indicate that MAP 1A and MAP 1B are structurally related components of the neuronal cytoskeleton with complementary patterns of expression.

Substance P and luteinizing hormone-releasing hormone levels in the brain of the male golden hamster are both altered by castration and testosterone replacement.

The effects of castration and testosterone (T) replacement on levels of substance P (SP) and luteinizing hormone-releasing hormone (LHRH) were assessed in discrete areas of the male hamster brain. The animals were either castrated, castrated and given a chronically low or high dose of T by Silastic implant, or sham-operated. Brain tissues and trunk blood were collected 3 weeks after surgery. Plasma T levels were maintained within the normal range by the implants but at significantly lower or higher levels than the mean for sham-operated males. Levels of SP and LHRH were quantified in the olfactory bulbs, rostral basal forebrain, anterior hypothalamic and preoptic area, medial basal hypothalamic area, medial basal hypothalamic area and median eminence, and brain stem. In general, castration and T replacement effected opposite changes in levels of SP and LHRH. In the medial basal hypothalamic area and median eminence SP levels were found to be inversely related to the chronic T levels, whereas the LHRH levels were directly correlated. In the anterior hypothalamic and preoptic area, castration reduced levels of SP. Conversely, castration elevated levels of LHRH in this area. This inverse dynamic relationship between changing peptide levels was also observed in the rostral basal forebrain but not in the olfactory bulbs. In most of these forebrain regions, the dose-response curves for the experimental groups could not incorporate the peptide levels in the sham-operated control group. SP levels in the brain stem showed a monotonic inverse relationship to circulating T levels which did include the control group values.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenovirus type 3 infection with systemic manifestation in apparently normal children.

Between July 1983 and February 1984, eight children with adenovirus Type 3 infection, proven by virus isolation from sputum, stool or nasopharyngeal swabs and a fourfold increase in complement fixation antibody titers against the virus, were treated in our department. All eight patients had fever lasting at least 7 days, hepatomegaly, diffuse pulmonary infiltrates and abnormal liver function tests. Seven of the patients exhibited dyspnea and pulmonary wheezing. Six of the patients developed changes in state of consciousness, and three had repeated convulsions. EEG patterns in three of the patients were compatible with encephalopathy. Other clinical manifestations included: follicular tonsillitis in two patients, diarrhea in two, pneumothorax in one, and shock with disseminated intravascular coagulation in one. The spectrum of adenovirus Type 3 infection reported here has been described previously only in the viral hemorrhagic fevers. This adenovirus Type 3 infection shares the potential for disseminated disease that has been described previously for Type 7, simulating Reye's syndrome.

Maturation of olfactory exploration in golden hamsters.

Following placement into a test cage filled with pine shavings, a litter of 7-8 golden hamster pups (aged 3-18 days postnatal: P3-18) initially displays a period of locomotion which ends reliably in huddling. The latency to establish a huddle (i.e., the duration of locomotion) is significantly longer in the presence of novel odors (fresh or lemon shavings) than more familiar odors (slightly soiled fresh or lemon shavings) but only in pups aged P12 or older. Pups aged P9 or younger do not locomote differentially in the presence of novel or familiar odors. This age difference represents the emergence of olfactory exploration in hamsters between P9 and P12. Exploration of novel odors interferes with initial attempts to establish a single huddle site by a litter, but does not preclude the ultimate aggregation of all pups at a single site as guided by conspecific odors and possibly thermotactile cues as well. Such shifts in the control of behavior by non-nest and nest-related, conspecific stimuli correspond well with the first occurrence of nest exits at P11-12 (e.g., Dieterlen, 1959) coupled with the persistent return of hamster pups to the maternal nest for as long as it is maintained (Rowell, 1961).

Precursor forms of substance P (SP) in nervous tissue: detection with antisera to SP, SP-Gly, and SP-Gly-Lys.

Antisera generated to substance P-Gly (SP-G) and substance P-Gly-Lys (SP-G-K), the likely unamidated COOH-terminally extended forms of substance P, were used to quantify and localize substance P precursor forms in hamster brain stem and spinal cord. The precursor determinant SP-G-K was liberated from larger heterogeneous forms by mild trypsinization of tissue extracts and was converted into the second precursor determinant, SP-G, by subsequent treatment with carboxypeptidase B. The basal levels of SP-G-K in brain stem and spinal cord were approximately equal to 0.5 pg/mg of tissue and rose 43- to 64-fold after trypsinization. Basal levels of SP-G were comparable to those of SP-G-K and rose 10- to 29-fold after combined enzyme treatments. Immunohistochemical labeling of axons and somata with anti-SP-G-K increased dramatically after trypsinization. This labeling was eliminated by preadsorption with authentic SP-G-K but not substance P or SP-G. Gel-permeation chromatography revealed SP-G-K-like immunoreactivity in fractions corresponding to considerably higher molecular weight than mature substance P. Collectively, these results support the hypothesis that substance P is synthesized from larger precursors and demonstrate that extended precursor forms are normally present in the axons and somata of neural systems that synthesize substance P.

Topographic organization of tufted cell axonal projections in the hamster main olfactory bulb: an intrabulbar associational system.

The organization of intrinsic axonal projections of principal neurons in the main olfactory bulb (MOB) was studied in hamsters by using wheat germ agglutinin-horseradish peroxidase (WGA-HRP) and fluorescent dyes. Punctate injections of either WGA-HRP or fast blue (FB) that are restricted to small sectors on one side of the MOB produce comparably restricted fields of retrograde labeling on the opposite side. Label is found predominantly in superficially situated (middle and external) tufted cells that lie near and at the border between the external plexiform and glomerular layers. Few of the deeper middle tufted, internal tufted, or mitral cells and no external tufted cells that lie in the superficial two-thirds of the glomerular layer are labeled in regions remote to the injection site. Anterograde transport of WGA-HRP from the injection site labels axons that travel dorsally and ventrally in restricted bands through the internal plexiform layer and then terminate within this layer in the punctate sector on the opposite side that contains retrogradely labeled neurons. Such reciprocal projections between opposing regions of the medial and lateral sides of the MOB are found at all rostrocaudal and dorsoventral levels. When punctate injections of FB into the MOB are paired with restricted injections of a second fluorescent tracer (nuclear yellow or diamidino yellow dihydrochloride) into the appropriate sector of pars externa (pE) of the anterior olfactory nucleus, the punctate region of remote retrogradely labeled principal neurons is embedded within a topographically restricted longitudinal wedge of retrogradely labeled mitral and tufted cells that project extrinsically to or through pE. However, extremely few of these neurons are double-retrogradely labeled. The results reveal the existence of an intrabulbar associational system in which principal neurons engage in point-to-point, reciprocal projections between opposing regions of the medial and lateral MOB. Moreover, the results indicate that this associational system largely arises from superficially situated tufted cells distinct from those that support bulbofugal projections into the topographically organized interbulbar commissural system via pE.

Topographic organization of connections between the main olfactory bulb and pars externa of the anterior olfactory nucleus in the hamster.

The organization of connections between the main olfactory bulb (MOB) and pars externa (pE) of the anterior olfactory nucleus was studied in hamsters by using wheat germ agglutinin-horseradish peroxidase as both an anterograde and a retrograde neuronal tracer. Bulbar efferents of pE project exclusively to the contralateral MOB. A topographic organization is evident in these efferents, such that distinct sectors of pE project predominantly to certain sectors of the contralateral MOB and lightly to other sectors. The predominant projection to any bulbar sector in the coronal plane is repeated at nearly all rostral-caudal levels, i.e., the pE efferents to the contralateral MOB terminate within long strips or wedges that show a sector-to-sector topographic organization with respect to the medial-lateral and dorsal-ventral axes but not the rostral-caudal axis of the MOB. Afferents to pE arising in the ipsilateral MOB also show a sector-to-sector topographic organization. Injections into restricted sectors along the circumference of pE label all classes of output neurons (mitral cells and internal, middle, and external tufted cells) in restricted sectors of the ipsilateral MOB, and the sectors that have retrograde neuronal labeling are homotopic to those in the contralateral MOB that have dense anterograde terminal labeling. External tufted cells are not labeled and the other classes of MOB output neurons do not have prominent topographic patterns of labeling in cases with injections caudal to pE. The somata of external tufted cell that project to pE are predominantly in the deep part of the glomerular layer; most of the external tufted cells that lie more superficially in the glomerular layer do not appear to have projections extrinsic to the MOB. These results indicate that both the afferent and efferent connections of pE with the MOB are topographically organized and provide a short synaptic pathway between homotopic sectors of the two main olfactory bulbs.

Widespread distribution of the major polypeptide component of MAP 1 (microtubule-associated protein 1) in the nervous system.

We prepared a monoclonal antibody to microtubule-associated protein 1 (MAP 1), one of the two major high molecular weight MAP found in microtubules isolated from brain tissue. We found that MAP 1 can be resolved by SDS PAGE into three electrophoretic bands, which we have designated MAP 1A, MAP 1B, and MAP 1C in order of increasing electrophoretic mobility. Our antibody recognized exclusively MAP 1A, the most abundant and largest MAP 1 polypeptide. To determine the distribution of MAP 1A in nervous system tissues and cells, we examined tissue sections from rat brain and spinal cord, as well as primary cultures of newborn rat brain by immunofluorescence microscopy. Anti-MAP 1A stained white matter and gray matter regions, while a polyclonal anti-MAP 2 antibody previously prepared in this laboratory stained only gray matter. This confirmed our earlier biochemical results, which indicated that MAP 1 is more uniformly distributed in brain tissue than MAP 2 (Vallee, R.B., 1982, J. Cell Biol., 92:435-442). To determine the identity of cells and cellular processes immunoreactive with anti-MAP 1A, we examined a variety of brain and spinal cord regions. Fibrous staining of white matter by anti-MAP 1A was generally observed. This was due in part to immunoreactivity of axons, as judged by examination of axonal fiber tracts in the cerebral cortex and of large myelinated axons in the spinal cord and in spinal nerve roots. Cells with the morphology of oligodendrocytes were brightly labeled in white matter. Intense staining of Purkinje cell dendrites in the cerebellar cortex and of the apical dendrites of pyramidal cells in the cerebral cortex was observed. By double-labeling with antibodies to MAP 1A and MAP 2, the presence of both MAP in identical dendrites and neuronal perikarya was found. In primary brain cell cultures anti-MAP 2 stained predominantly cells of neuronal morphology. In contrast, anti-MAP 1A stained nearly all cells. Included among these were neurons, oligodendrocytes and astrocytes as determined by double-labeling with anti-MAP 1A in combination with antibody to MAP 2, myelin basic protein or glial fibrillary acidic protein, respectively. These results indicate that in contrast to MAP 2, which is specifically enriched in dendrites and perikarya of neurons, MAP 1A is widely distributed in the nervous system.