Efficient retrovirus-mediated transfer of the multidrug resistance 1 gene into autologous human long-term repopulating hematopoietic stem cells.

Pre-clinical studies indicate that efficient retrovirus-mediated gene transfer into hematopoietic stem cells and progenitor cells can be achieved by co-localizing retroviral particles and target cells on specific adhesion domains of fibronectin. In this pilot study, we used this technique to transfer the human multidrug resistance 1 gene into stem and progenitor cells of patients with germ cell tumors undergoing autologous transplantation. There was efficient gene transfer into stem and progenitor cells in the presence of recombinant fibronectin fragment CH-296. The infusion of these cells was associated with no harmful effects and led to prompt hematopoietic recovery. There was in vivo vector expression, but it may have been limited by the high rate of aberrant splicing of the multidrug resistance 1 gene in the vector. Gene marking has persisted more than a year at levels higher than previously reported in humans.

Difference in brain densities between chronic alcoholic and normal control patients.

The densities of the brains of 11 chronic alcoholics were compared with those of 11 age-matched normal control subjects. Densities were determined from the density numbers generated by computerized tomography at three levels of the brain-the highest level of the lateral ventricles and the next two higher levels-with adjustments made to control for possible artifacts in the data. The advantage of the dominant hemisphere over the nondominant hemisphere was lessened in alcoholics.

Interprofessional provider attitudes toward the initiation of epidural analgesia in the laboring patient: are we all on the same page?

BACKGROUND: The timing of initiation of neuraxial labor analgesia should ultimately depend on patient preference although obstetricians, anesthesiologists and nurses may influence decision-making. We hypothesized that provider groups would have similar attitudes toward the timing of epidural placement, but some identifiable differences could be used to improve understanding and communication among providers. METHODS: Anesthesiologists, nurses and obstetricians completed a survey assessing their knowledge and attitudes on the timing of epidural placement in specified clinical circumstances. RESULTS: Anesthesiologists (100%) and nurses (86.2%) reported being more familiar with epidural management than obstetricians (43.3%, P <0.01). The willingness of providers to advocate epidural placement based on the magnitude of cervical dilation was similar, although at 10 cm dilatation obstetricians (73.3%) were significantly more likely to advocate neuraxial block compared to both nurses (27.6%, P <0.01) and anesthesiologists (36.7%, P <0.01). The impact of patient factors and clinical circumstances on the timing of neuraxial block placement showed significant differences among provider groups in five of 24 areas assessed, including patient desire for an epidural, primigravid patients without membrane rupture, oxytocin infusion initiated, labor epidural in a previous pregnancy, and a difficult airway. CONCLUSIONS: There were differences between providers in factors that may impact the timing of epidural placement and in their self-perceived familiarity with epidural management. These present an opportunity for furthering interprofessional education and collaboration.

Measuring Gratitude in Children.

Gratitude is a rich socioemotional construct that emerges over development beginning in early childhood. Existing measures of children's gratitude as a trait or behavior may be limited because they do not capture different aspects of gratitude moments (i.e., awareness, thoughts, feelings, and actions) and the way that these facets appear in children. The current study evaluates a battery of new measures assessing children's gratitude to address these limitations. Parent-child dyads (N=101; children aged 6-9) completed a lab-based assessment followed by a 7-day online parental diary and 18-month follow-up survey. In addition to newly developed measures of children's gratitude, the battery included indicators of convergent, concurrent, divergent, and predictive validity. Results demonstrate the complexity of gratitude as a construct and the relative benefits and limits of various assessment modalities. Implications for the measurement of children's gratitude and suggestions for future research on the development of gratitude are discussed.

Anesthetic management of parturients with Arnold Chiari malformation-I: a multicenter retrospective study.

BACKGROUND: Consensus regarding the safest mode of delivery and anesthetic management for parturients with Arnold Chiari malformation-I (ACM-I) remains controversial. This study assessed their anesthetic management and reported anesthetic complications during hospitalization for delivery. METHODS: This was a multicenter, retrospective, cohort study of patients with ACM-I undergoing vaginal or cesarean delivery. Data were obtained from the electronic databases of four United States academic institutions using International Classification of Diseases (ICD) codes from 2007-2017 at three sites and 2004-2017 at one site. The primary outcome was anesthetic complications. RESULTS: Data were analyzed for 185 deliveries in 148 patients. Diagnosis of ACM-I was made prior to delivery in 147 (80%) cases. Pre-delivery neurosurgical consultation for management of ACM-I was performed in 53 (36%) patients. Pre-existing symptoms were recorded for 89 (48%) of the deliveries. Vaginal deliveries occurred in 80 (43%) cases, and 62 women (78%) received neuraxial labor analgesia. Cesarean delivery was performed in 105 (57%) cases, of which 70 women (67%) had neuraxial anesthesia and 34 (32%) received general anesthesia. Post-dural puncture headache was reported in three (2%) patients who had neuraxial anesthesia, and in two (12%) patients with syringomyelia. There was one (3%) reported case of aspiration pneumonia with general anesthesia. CONCLUSIONS: The findings suggest that anesthetic complications occur infrequently in patients with ACM-I regardless of the anesthetic management. Although institutional preference in anesthetic and obstetric care appears to drive patient management, the findings suggest that an individualized approach has favorable outcomes in this population.

BMI-based body size guides for women and men: development and validation of a novel pictorial method to assess weight-related concepts.

OBJECTIVES: To develop and evaluate two BMI-based instruments to determine perceptions of weight status, particularly perceptions of overweight and obesity, using pictorial images of women and men. METHODS: Pictures of adults with known BMI values were used to construct gender-specific body size guides (BSGs) containing 10 bodies that ranged from underweight to class III obesity. Figures were standardized and a composite face was added to each. The BSGs were administered to 400 adults to assess the psychometric properties of the instruments and weight perceptions. RESULTS: High correlations between the BMIs of respondents and the BMIs of the current body selected by respondents provided strong support for the criterion-related validity of the BSGs, and the logical pattern of responses to items assessing perception of weight categories supported construct validity for the scales. Test-retest reliability, assessed by correlations for both current and ideal body, was also high, despite the lengthy 6-month testing interval. Respondents' perceptions of the bodies within specific weight categories indicated that a majority failed to recognize the overweight female as overweight and perceived the overweight male as normal weight. Obese bodies were generally unrecognized as such until the bodies reached the higher levels of obesity (that is, BMI values >39). Perception of weight was influenced by the respondents' weight status and gender. CONCLUSIONS: Psychometric analyses indicated the BSGs are valid and reliable instruments. These results, coupled with the face validity of the scales and the relationship between the bodies and BMI values, indicate the BSGs offer advantages over existing instruments for researchers of weight perception and body image. Administration of the scales to an adult sample confirmed that overweight and obesity are under-recognized. Increased efforts to improve public understanding of these terms are needed and the BSGs may provide useful tools for this purpose.

Expression of spatially regulated genes in the sea urchin embryo.

Spatially controlled genes expressed in the early sea urchin embryo have been characterized, and the patterns of expression in terms of the mechanisms by which this embryo accomplishes its initial set of founder cell specifications are the subject of current discussion. Sea urchin transcription factors that have been cloned are classified with respect to their target sites and the genes they regulate. Among the best known of the sea urchin cis-regulatory systems is that controlling expression of the Cyllla gene, which encodes an aboral ectoderm-specific cytoskeletal actin. The Cyllla regulatory domain includes approximately 20 sites of DNA-protein interaction, serviced by about ten different factors. Certain of these factors are known to negatively control spatial expression, while others positively regulate temporal activation and the level of Cyllla gene expression. Differential, lineage-specific gene expression is instituted in the sea urchin embryo by mid-late cleavage, prior to any cell migration or overt differentiation, and shortly following lineage segregation.

Combinatorial regulation of the Saccharomyces cerevisiae CAR1 (arginase) promoter in response to multiple environmental signals.

CAR1 (arginase) gene expression responds to multiple environmental signals; expression is induced in response to the intracellular accumulation of arginine and repressed when readily transported and catabolized nitrogen sources are available in the environment. Up to 14 cis-acting sites and 9 trans-acting factors have been implicated in regulated CAR1 transcription. In all but one case, the sites are redundant. To test whether these sites actually participate in CAR1 expression, each class of sites was inactivated by substitution mutations that retained the native spacing of the CAR1 cis-acting elements. Three types of sites function independently of the nitrogen source: two clusters of Abflp- and Rap1p-binding sites, and a GC-rich sequence. Two different sets of nitrogen source-dependent sites are also required: the first consists of two GATAA-containing UASNTR sites that mediate nitrogen catabolite repression-sensitive transcription, and the second is arginine dependent and consists of three UAS1 elements that activate transcription only when arginine is present. A single URS1 site mediates repression of CAR1 arginine-independent upstream activator site (UAS) activity in the absence of arginine and the presence of a poor nitrogen source (a condition under which the inducer-independent Gln3p can function in association with the UASNTR sites). When arginine is present, the combined activity of the UAS elements overcomes the negative effects mediated by URS1. Mutation of the classes of sites either singly or in combination markedly alters CAR1 promoter operation and control, supporting the idea that they function synergistically to regulate expression of the gene.

Gat1p, a GATA family protein whose production is sensitive to nitrogen catabolite repression, participates in transcriptional activation of nitrogen-catabolic genes in Saccharomyces cerevisiae.

Saccharomyces cerevisiae cells selectively use nitrogen sources in their environment. Nitrogen catabolite repression (NCR) is the basis of this selectivity. Until recently NCR was thought to be accomplished exclusively through the negative regulation of Gln3p function by Ure2p. The demonstration that NCR-sensitive expression of multiple nitrogen-catabolic genes occurs in a gln3 delta ure2 delta dal80::hisG triple mutant indicated that the prevailing view of the nitrogen regulatory circuit was in need of revision; additional components clearly existed. Here we demonstrate that another positive regulator, designated Gat1p, participates in the transcription of NCR-sensitive genes and is able to weakly activate transcription when tethered upstream of a reporter gene devoid of upstream activation sequence elements. Expression of GAT1 is shown to be NCR sensitive, partially Gln3p dependent, and Dal80p regulated. In agreement with this pattern of regulation, we also demonstrate the existence of Gln3p and Dal80p binding sites upstream of GAT1.

A GBF-binding site and a novel AT element define the minimal sequences sufficient to direct prespore-specific expression in Dictyostelium discoideum.

In order to better understand the molecular mechanisms of cellular differentiation in Dictyostelium discoideum, we have identified the minimum regulatory sequences of the prespore-specific gene SP60/cotC that are sufficient to confer cell-type-specific expression on a heterologous promoter. This region includes at least two essential cis-acting elements: a novel AT-rich element (or elements) and CAE3. The essential function of the AT element is confirmed through point mutations that decrease expression below the level of detection. CAE3 is one of three CA-rich elements (CAEs) required for the induction of SP60/cotC during development or in response to extracellular cyclic AMP. The CAEs have differential affinities for a specific developmentally induced nuclear activity (CAE1 > CAE2 >> CAE3). Here, we identify this activity as G-box-binding factor (GBF) and show that in vitro-transcribed and -translated GBF binds all three SP60/cotC CAEs in a sequence-specific manner. Previous studies have suggested that GBF mediates the induction of some prestalk genes, and these results demonstrate that it also has a specific role in prespore gene activation.

Spatial and temporal expression of the Dictyostelium discoideum G alpha protein subunit G alpha 2: expression of a dominant negative protein inhibits proper prestalk to stalk differentiation.

Previous results have shown that the G alpha protein subunit G alpha 2 is required for aggregation in Dictyostelium discoideum and is essential for coupling cell-surface cAMP receptors to downstream effectors in vivo during this stage of development. G alpha 2 expresses at least four distinct transcripts that are differentially regulated during development; two of the transcripts are expressed exclusively in the multicellular stages and their expression is restricted to prestalk cells. We partially dissected the G alpha 2 promoter and identified a component that is expressed exclusively during the multicellular stages using luciferase gene fusions. When this promoter region is coupled to lacZ, beta-gal expression is restricted to the multicellular stages and localized in prestalk cells with a pattern similar to that of the ecmA prestalk-specific promoter. We show that expression in wild-type cells of the G alpha 2 mutant protein [G alpha 2(G206T)] during the early stages of development blocks aggregation and cAMP-mediated activation of adenylyl cyclase and guanylyl cyclase, suggesting it functions as a dominant negatively active G alpha subunit. When this mutant G alpha protein is expressed from the ecmA prestalk-specific promoter, abnormal stalk differentiation during culmination is observed. Expression of the mutant G alpha 2 from the SP60 prespore promoter or wild-type G alpha 2 from either the ecmA or the SP60 promoter results in no detectable phenotype. The results suggest that G alpha 2 plays an essential role during the culmination stage in prestalk cells and may mediate cAMP receptor activation of these processes during multicellular development.

Transversus abdominis plane and ilioinguinal/iliohypogastric blocks for cesarean delivery in a patient with type II spinal muscular atrophy.

While neuraxial and general anesthetic techniques are most commonly utilized for cesarean delivery, there are rare instances in which alternative techniques may be considered. We report a patient with type II spinal muscular atrophy who had relative contraindications to both neuraxial and general anesthesia, and had experienced significant discomfort during two previous cesarean deliveries performed with local anesthetic infiltration. We describe the successful use of bilateral ultrasound-guided transversus abdominis plane and ilioinguinal/iliohypogastric blocks, in addition to intravenous sedation, for cesarean delivery anesthesia.

Structural brain deficits in schizophrenia. Identification by computed tomographic scan density measurements.

Research has suggested the presence of brain damage as a cause or concomitant of chronic schizophrenia. The most recent research in this area has been the identification of abnormalities in schizophrenia by computed tomographic (CT) scans. A study was done to investigate localized changes in CT scan density numbers in the brains of schizophrenic patients, as opposed to the brains in normal control subjects. Twenty-four normal subjects and 23 schizophrenic patients were tested with CT scans. Density measurements in each area of the brain (left, right, anterior and posterior) were compared to three separate CT scan levels. Of six measurements of anterior left-hemisphere density, it was found that five showed lower density in schizophrenic brains, as compared with normal brains. Of the remaining 18 measurements that evaluated other areas of the brain, only three differentiated between schizophrenic patients and normal subjects. The results support the hypothesis that there are primary structural deficits in some schizophrenic patients, and these deficits are centered in and around the anterior area of the left (dominant) hemisphere. The results also demonstrated further implications.

Structural abnormalities in the frontal system in schizophrenia. A magnetic resonance imaging study.

Thirty-eight schizophrenics and 49 normal controls underwent magnetic resonance imaging. Midline sagittal cuts indicated that the schizophrenics had significantly smaller frontal lobes, as well as smaller cerebrums and craniums. The findings are consistent with some type of early developmental abnormality that might retard brain growth and therefore skull growth. These findings are confirmed on a smaller sample of patients on whom we have coronal cuts. Decreased cerebral and cranial size are associated with prominent negative symptoms, although decreased frontal size is not. Decreased cranial and cerebral size was also associated with impairment on some cognitive tests. These findings are consistent with the hypothesis that some schizophrenics may have a type of early developmental abnormality associated with prominent negative symptoms and cognitive impairment. Further, the results suggest that schizophrenics may have a type of structural frontal system impairment. Thus, they provide anatomic evidence for the "hypofrontality hypothesis."

Digital vasodilatation during mental stress in patients with Raynaud's disease.

Fingertip blood flow was measured by venous occlusion plethysmography before and during a stressful mental task consisting of rapid serial arithmetic calculations in a 25 degrees C room. Significant rises in heart rate and blood pressure indicated that stress was actually induced in all individuals. During mental stress in normal subjects, blood flow decreased (46.4 +/- 6.2 to 22.4 +/- 4.9 ml X min-1 per 100 ml tissue; P less than 0.01) and vascular resistance increased (2.1 +/- 0.4 to 7.6 +/- 2.2 units; P less than 0.01). Patients with Raynaud's disease unexpectedly increased blood flow (15.4 +/- 4.2 to 21.6 +/- 5.7; P = 0.05) and decreased vascular resistance (9.7 +/- 2.3 to 7.1 +/- 1.4; P = 0.05). Ten additional normal subjects were studied in a cool room (20 degrees C). Their digits remained vasoconstricted during stress, as blood flow (7.4 +/- 2.9 to 5.1 +/- 1.3) and vascular resistance (31.5 +/- 11.1 to 34.4 +/- 8.2) varied insignificantly (P greater than 0.10). The digital vasodilatation which occurs during mental stress in patients with Raynaud's disease was not altered by pretreatment with oral indomethacin, with intra-arterial propranolol or atropine, or by digital nerve block. These findings suggest the existence of an active digital vasodilatory mechanism in patients with Raynaud's disease.

Raynaud's phenomenon. An update.

The pathogenesis of primary Raynaud's phenomenon remains an enigma. Most evidence favors a local abnormality in the digital arteries as opposed to an increased activity of the sympathetic nervous system. The local fault may involve the alpha 2-adrenergic receptors, which are most important in reflex sympathetic vasoconstriction. Cooling blood vessels increase the sensitivity of alpha 2-adrenergic receptors, increased levels of alpha 2-adrenergic receptors are present in primary Raynaud's disease, and patients show an increased sensitivity to alpha 2-adrenergic receptor agonists on finger blood flow. Serotonin has also been implicated, but the evidence is not compelling. In secondary Raynaud's phenomenon, vasospastic attacks can often be explained by a low arterial distending pressure, a thickened vessel wall, or absence of beta-adrenergic receptor activity. Diagnosis of primary Raynaud's disease relies on a typical history and normal physical examination, laboratory studies, and nailfold capillaroscopy. Finger systolic blood pressures during local cooling with ischemia may be helpful to document vasospastic attacks but does not distinguish primary from secondary Raynaud's phenomenon. The treatment of Raynaud's phenomenon is usually conservative. Pavlovian conditioning or biofeedback may be beneficial. When drug therapy is necessary, the calcium channel entry blocker nifedipine or sympatholytic agents have been shown to decrease the frequency and duration of vasospastic attacks in about two thirds of patients, although subjective improvement does not usually correlate with objective testing. Direct-acting vasodilators have not been shown to be of definite benefit. New therapies include prostaglandins, captopril, and the serotonergic antagonist ketanserin. Surgical sympathectomy has not been beneficial.

Pharmacological concentrations of ascorbic acid are required for the beneficial effect on endothelial vasomotor function in hypertension.

Increased production of superoxide anion may contribute to impaired bioactivity of endothelium-derived nitric oxide in hypertension. Ascorbic acid is capable of scavenging superoxide anion; however, experimental studies have shown that high physiological concentrations (>1 mmol/L) of ascorbic acid are required to prevent superoxide-mediated vascular dysfunction. To seek kinetic evidence that superoxide anion contributes to endothelial vasomotor dysfunction in human hypertension, we examined the effects of 2.4 or 24 mg/min ascorbic acid intra-arterial infusions on forearm blood flow responses to methacholine or sodium nitroprusside in 30 patients with hypertension and 22 age-matched controls. Endothelium-dependent vasodilation to methacholine was significantly impaired in the hypertensive patients, with a response to the highest dose of methacholine (10 microg/min) of 12.3+/-6.7 compared with 16.1+/-5.8 mL. min(-1). dL tissue(-1) in the controls (P<0.001). The response to sodium nitroprusside was equivalent in the 2 groups. Ascorbic acid at 24 mg/min significantly improved the forearm blood flow response to methacholine in hypertensive patients with a peak response of 16.1+/-7.1 mL. min(-1). dL tissue(-1) (P=0.001). This dose produced a cephalic vein ascorbic acid concentration of 3.2+/-1. 4 mmol/L. In contrast, ascorbic acid at 2.4 mg/min had no effect on the methacholine response. Ascorbic acid at both doses had no effect on the vasodilator response to sodium nitroprusside in hypertensive patients or the methacholine response in the controls. These results agree with the predicted kinetics for superoxide anion-mediated impairment of endothelium-derived nitric oxide action. Thus, superoxide anion may contribute to impaired endothelium-dependent vasodilation in patients with hypertension.

Left hemispheric density deficits in chronic schizophrenia.

Regional determinations of mean cerebral tissue density were made for 50 chronic schizophrenics and compared with results in 24 normal controls. When values for corresponding left and right hemispheric regions were compared, 11 of 12 comparisons showed the left regional value to be significantly greater than the right for controls while only 6 of 12 comparisons among schizophrenics showed left values significantly greater than right. These data suggest an overall decreased relative density of the left cerebral hemisphere in schizophrenia. Further implications of the data are discussed.

Perinatal brain injury and cerebellar vermal lobules I-X in schizophrenia.

Several studies, including our own, have reported atrophy of the cerebellar vermis in some schizophrenic patients. A recent report by Courchesne et al (1988) of hypoplasia of a developmentally specific region of the cerebellar vermis in autism prompted us to hypothesize that the cerebellar "atrophy" in some schizophrenic patients may also have developmental origins. We measured the area of the vermal lobules in 30 male schizophrenics. Contrary to expectation, the patients as a group had consistently larger cerebellar structures than the controls. Patients with perinatal injury had smaller structures than the nonperinatally injured group, but these measures were still larger than in the control subjects. Patients without perinatal injury differed from controls, having larger lobules VI-VII (p less than 0.03). These preliminary findings tentatively suggest a role for developmental factors for cerebellar structures in schizophrenia. Further research is needed to clarify the cerebellar vermal changes observed in some schizophrenic patients.