High prevalence of adrenal insufficiency in patients with sickle cell disease: results from a community hospital in the U.S.

OBJECTIVE: Despite the low prevalence (0.008%) of adrenal insufficiency (AI) in the general population, this disorder was recently diagnosed in a substantial number of sickle cell disease (SCD) patients at our hospital. The main objective of this study was to assess the prevalence of AI in SCD patients. METHODS: All adult patients admitted to the Department of Medicine at Interfaith Medical Center from October 2010 to November 2011 were eligible for this retrospective study. Medical records of adult SCD patients hospitalized for painful crisis and who had undergone cosyntropin testing were reviewed. Adult non-SCD patients hospitalized for painful crisis and who had undergone cosyntropin testing served as controls. The result of the cosyntropin test was the primary outcome. The prevalence of positive cosyntropin tests was compared between the 2 groups by using Student's t-test, and odds ratios. RESULTS: 62 adult SCD patients were enrolled in the study. 15 underwent cosyntropin testing and 12 (19.4%) of these patients were found to have AI. AI was also diagnosed in 1 of 1,340 non-SCD patients. The odds ratio for AI in SCD to non-SCD patients [(12/62)/(1,340)] was 259. The odds ratio for the prevalence of AI in SCD patients in our study (19.4%) vs. the general population (approximately 0.008%) was 2,375. CONCLUSION: AI occurred in 19.4% of SCD patients included in this study. These patients thus have a 2,375-fold higher risk of developing AI than the general population, and a 259-fold greater risk of developing AI than do hospitalized non-SCD patients.

Interleukin-17 stimulates chemokine (interleukin-8 and monocyte chemoattractant protein-1) secretion in human pancreatic periacinar myofibroblasts.

BACKGROUND: Interleukin (IL)-17 is a newly identified T-cell-derived cytokine that can regulate the functions of a variety of cell types. In this study, we investigated the effects of CD4+ T-cell-derived cytokines on chemokine secretion in human pancreatic periacinar myofibroblasts. METHODS: The secretion of IL-8 and monocyte chemoattractant protein (MCP)-1 was evaluated by ELISA and Northern blot. The expression of IL-17 receptor (R) was analyzed by Northern blot and a binding assay using 125I-labeled IL-17. The activation of nuclear factor-kappaB (NF-kappaB) was assessed by an electrophoretic gel mobility shift assay (EMSA). RESULTS: IL-17 induced a dose-dependent increase in IL-8 and MCP-1 secretion. The effects of IL-17 on IL-8 and MCP-1 mRNA abundance reached a maximum as early as 3 h. and then gradually decreased. IL-17 and IFN-gamma synergistically increased IL-8 secretion and additively enhanced MCP-1 secretion. IFN-gamma induced a weak increase in IL-17R mRNA abundance, but incubation with IFN-gamma for 24 h had no effects on 125I-labeled IL-17-binding, indicating that the co-stimulatory effects of IL-17 and IFN-gamma were not regulated by the modulation of IL-17R expression. Furthermore, IL-17 induced a rapid increase in NF-kappaB DNA-binding activity, and the combination of IL-17 and IFN-gamma further enhanced NF-kappaB DNA-binding activity. CONCLUSIONS: In conclusion, it becomes clear that IL-17 is an inducer of IL-8 and MCP-1 secretion in human pancreatic periacinar myofibroblasts. The combination of IL-17 with IFN-gamma further enhances chemokine secretion. These findings indicate a linkage between T-cell-mediated immunity and inflammatory responses in the pancreas.

Ultrasonic vocalization responses in genetically high- and low-emotional rats.

Tsukuba High Emotional (THE) and Tsukuba Low Emotional (TLE) strains of rats have been congenitally bred for use in studies of emotionality. The current study investigated THE and TLE strain differences in the footshock-induced ultrasonic vocalization responses of adult male rats. Ultrasonic vocalization response inducibility (ratio of rats emitting ultrasounds) and vocalization activity in THE rats were statistically higher than in TLE rats. We next examined the causal relation between the ultrasonic vocalizations and the activation of the pituitary-adrenocortical axis in response to footshocks in these two strains of rats. Rats were sorted into two groups, vocalizing and non-vocalizing, after being exposed to the shock regimen daily for 5 successive days. Basal plasma concentrations of ACTH and corticosterone were not different between THE and TLE rats. After receiving footshocks, significant increases in plasma concentrations of ACTH and corticosterone were induced in both strains of rats. These increases in plasma concentrations of ACTH and corticosterone were significantly higher in THE than in TLE rats. However, in vocalizing and non-vocalizing rats of both strains, no statistical differences in plasma concentrations of ACTH and corticosterone were observed after footshocks. These findings suggest that the high emotionality of the adult male rats was reflected in the emission of ultrasounds, and that the emission of ultrasonic vocalizations might not be related to the activation of the pituitary-adrenocortical axis.

Neurosteroid synthesis by cytochrome p450-containing systems localized in the rat brain hippocampal neurons: N-methyl-D-aspartate and calcium-dependent synthesis.

Neurosteroidogenesis has not been well elucidated due to the very low level of steroidogenic proteins in the brain. Here we report the first demonstration of the neuronal localization of neurosteroidogenic systems as well as the regulation of neurosteroidogenic activity in the adult rat hippocampus. Significant localization of cytochrome P450scc was observed in pyramidal neurons and granule neurons by means of immunohistochemical staining of slices. We also observed the colocalization, in hippocampal neurons, of P450scc with redox partners, hydroxysteroid sulfotransferase and steroidogenic acute regulatory protein. The distributions of astroglial cells and oligodendroglial cells showed very different patterns from that of the P450scc-containing cells. The expression of P450scc, redox partners, the sulfotransferase, and steroidogenic acute regulatory protein was also confirmed by Western blot analysis. The process of active neurosteroidogenesis was stimulated by exposing neurons to N-methyl-D-aspartate. Upon stimulation with N-methyl-D-aspartate, Ca(2+) influx through the N-methyl-D-aspartate subtype of glutamate receptors occurred, and significant net production of pregnenolone and pregnenolone sulfate was observed in the hippocampus. This neurosteroid production was considerably suppressed by the addition of antagonists of N-methyl-D-aspartate receptors, by Ca(2+) depletion, or by the addition of an inhibitor of P450scc. Upon stimulation with N-methyl-D-aspartate, the processing of full-length steroidogenic acute regulatory protein (37-kDa) to the truncated 30-kDa steroidogenic acute regulatory protein was observed. Taken together, these observations imply that hippocampal neurons synthesize neurosteroids. This synthesis may be stimulated and regulated by glutamate-mediated synaptic communication.

Cooperation of interleukin-17 and interferon-gamma on chemokine secretion in human fetal intestinal epithelial cells.

Interleukin (IL)-17 is a newly identified T cell-derived cytokine that can regulate the functions of a variety of cell types. In this study, we investigated the effects of IL-17 and interferon (IFN)-gamma on chemokine secretion in human fetal intestinal epithelial cells. IL-8 and monocyte chemoattractant protein (MCP)-1 secretion by the human fetal intestinal epithelial cell line, intestine-407, was evaluated by ELISA and Northern blot. The expression of IL-17 receptor (R) was analysed by a binding assay using [(125)I]-labelled IL-17. The activation of nuclear factor-kappa B (NF-kappa B), NF-IL6 and AP-1 was assessed by an electrophoretic gel mobility shift assay (EMSA). IL-17 induced a dose-dependent increase in IL-8 and MCP-1 secretion. The inducing effects of IL-17 on IL-8 and MCP-1 mRNA abundance reached a maximum as early as 3 h, and then gradually decreased. IL-17 and IFN-gamma synergistically increased IL-8 and MCP-1 secretion and mRNA abundance. IFN-gamma induced a weak increase in IL-17 R mRNA abundance, and incubation with IFN-gamma for 24 h enhanced [(125)I]-labelled IL-17-binding by 2.4-fold. IL-17 rapidly induced the phosphorylation and degradation of I kappa B alpha molecules, and the combination of IL-17 and IFN-gamma induced a marked increase in NF-kappa B DNA-binding activity as early as 1.5 h after the stimulation. Furthermore, this combination induced an increase in NF-IL-6 and AP-1 DNA-binding activity. In conclusion, it becomes clear that IL-17 is an inducer of IL-8 and MCP-1 secretion by human fetal intestinal epithelial cells. The combination of IL-17 with IFN-gamma synergistically enhanced chemokine secretion. These effects of IL-17 and IFN-gamma might play an important role in the inflammatory responses in the intestinal mucosa.

Application of 2-aminopyridine fluorescence labeling in the analysis of in vivo and in vitro metabolism of dextran sulfate sodium by size-exclusion high-performance liquid chromatography.

The present study describes a size-exclusion high-performance liquid chromatographic method for the separation and quantification of sulfated polysaccharides, such as dextran sulfate sodium (DSS). Pyridylamination of DSS was achieved without difficulty using 2-aminopyridine as a fluorometric label. In addition, 0.1-0.2 M phosphate buffer (pH 3.0) was found to be the mobile phase which produced the best separation. In vitro enzymatic degradation of the pyridylamino-DSS (PA-DSS5000, Mr 5000) using alpha-amylase and the in vivo metabolism in the rat feces after oral administration of PA-DSS5000 were then evaluated. Two small peaks of approximately Mr 380 and 600 appeared after co-incubation with alpha-amylase, indicating PA-DSS5000 may be considerably depolymerized. In vivo, however, PA-DSS5000 excreted in the feces was mainly of PA-DSS5000 polymer. No peaks of less than Mr 5000 were not clearly detectable in the feces because of background fluorescence attributable to gut lumen contents. This method of fluorometric analysis allows fairly selective detection of sulfated polysaccharides in biological materials.

Ultrasonic isolation calls in genetically high- and low-emotional rat pups.

The present study was undertaken to investigate the relationship between the emotionality and the modulation of ultrasonic vocalizations in Tsukuba High Emotional (THE) and Tsukuba Low Emotional (TLE) strain rat pups 3-18 days old. The THE pups, while isolated from their dam and littermates and placed in a cold environment, emitted ultrasonic isolation calls at a high rate until day 15. In contrast, ultrasound production was at a consistently low level throughout the test period in the TLE pups. The ultrasonic isolation calls of THE pups were attenuated to the same level as those of the TLE pups after administration of diazepam (1 mg/kg, s.c.), a benzodiazepine receptor agonist, at 6 and 12 days of age. These findings suggest that the high emotionality of the rat pups was reflected largely by the emission of ultrasounds in response to isolation distress rather than the number of the benzodiazepine receptors in the brain that might play a role in physiologic mediation of the rat pup isolation call.

Efficacy of oral adsorbent for treatment of peristomal fistula associated with Crohn's disease.

Perianal fistulae are frequently seen complications in Crohn's disease. Although surgical procedures such as Seton's method have been devised, many patients still suffer from fistulae that are resistant to conventional therapy. We administered oral adsorbent to a patient with disease Crohn's who had a complicated peristomal fistula that did not improve with conventional therapy. Six grams of oral adsorbent (AST-120) were added daily to a regimen of elemental diet therapy and prednisolone. The fistula gradually decreased in size after the administration of the oral adsorbent, and had healed completely after 40 days' treatment. There were no side effects from the oral adsorbent. This case report suggests that oral adsorbent is an effective treatment for peristomal fistula associated with Crohn's disease.

On the Cusp around Central Black Holes in Luminous Elliptical Galaxies.

In this Letter, we show that a massive black hole (MBH) that falls into the center of a galaxy on the dynamical timescale leaves a weak cusp (rho~r-1&solm0;2) around it, which is in good agreement with the recent observations of luminous elliptical galaxies by the Hubble Space Telescope. Such an event is a natural outcome of the merging of two galaxies that have central MBHs. This is the only known mechanism for forming weak cusps in luminous elliptical galaxies. Therefore, the existence of the weak cusps indicates that the central black holes of luminous elliptical galaxies have fallen to the center from outside, most likely during a major merger event.

Astrophysics on the GRAPE family of special-purpose computers

The GRAPE-4, the world's fastest computer in 1995-1997, has produced some major scientific results through a wide diversity of large-scale simulations in astrophysics. Applications have included planetary formation, the evolution of star clusters and galactic nuclei, and the formation of galaxies and clusters of galaxies.

An integrated multimedia medical information network system.

An integrated multimedia medical information network system at Shimane Medical university has been developed to organize medical information generated from each section and provide information services useful for education, research and clinical practice. The report describes the outline of our system. It is designed to serve as a distributed database for electronic medical records and images. We are developing the MML engine that is to be linked to the world wide web (WWW) network system. To the users, this system will present an integrated multimedia representation of the patient records, providing access to both the image and text-based data required for an effective clinical decision making and medical education.

Defensive burying in two strains of rats selected for emotional reactivity.

This paper examined defensive burying in Tsukuba High- and Low-Emotional rats (THE and TLE). The THE rats showed significantly more defensive burying and more immobility than the TLE rats. The TLE rats were more active and started prod-investigation faster than the THE rats. Some TLE rats bit the prod immediately after shock administration. No significant strain differences were observed in passive avoidance (stay away from the prod) and time spent at prod-investigation. The results indicate that THE rats are active and TLE rats are inactive in relation to defensive burying, although THE rats generally show passive coping. The relation between defensive burying and strain-specific coping styles is discussed.

Role of angiotensin II in the transforming growth factor-beta 1 expression of rat kidney in anti-glomerular basement membrane antiserum-induced glomerulonephritis.

Induction of acute nephritis in the rat by injecting anti-glomerular basement membrane (GBM) antiserum is accompanied by a transient increase in angiotensin II generation in blood circulation within the first 24 h and a subsequent elevation of transforming growth factor-beta 1 (TGF-beta 1) mRNA levels in kidney cortex with a peak at days 7-8. Studies were carried out to determine whether the increased generation of angiotensin II plays a role in the elevation of TGF-beta 1 mRNA. Elevation of TGF-beta 1 mRNA levels 7 d after injection of antiserum was significantly inhibited by a successive daily administration of TCV-116, angiotensin II type 1 receptor antagonist, at 1 mg/kg/d from days 0 to 2 or from days 0 to 6, while it was not influenced by a single administration of this dose on day 0. In addition, angiotensin II infusion or 24 h at a rate of 50 ng/min did not alter the level of TGF-beta1 mRNA which was measured 6 d after the infusion. These results suggest that the anti-GBM antiserum-induced increase in TGF-beta 1 expression in the kidney is not responsible for angiotensin II generated in the blood circulation during the early phase of acute nephritis, but iis probably mediated by angiotensin II generated locally in the kidney.

A special-purpose computer for molecular dynamics: GRAPE-2A.

Molecular dynamics simulations have been extensively used in research of proteins. Since these simulations are quite computer intensive, their acceleration is of main interest of the research. In molecular dynamics simulations, almost all computing time is consumed in calculating the forces between particles, e.g., Coulomb and van der Waals forces. We have designed and built GRAPE-2A (GRAvity PipE 2A), a special-purpose computer for use in simulations of classical many-body systems. GRAPE-2A calculates forces exerted on a particle from the other particles. GRAPE-2A can calculate force of an arbitrary functional form of a central force. The host computer, which is connected to GRAPE-2A through the VME bus, performs other calculations such as time integration. The peak speed of GRAPE-2A is 180 Mflops. We can also stimulate systems with periodic boundary conditions by the Ewald method, using GRAPE-2A and another special-purpose computer, WINE (Wave space INtegrator for the Ewald method).

[Response types to shock and avoidance learning in inbred strains of mice].

The effects of shock intensity on response types to shock (Experiment 1) and the relationship between response types and avoidance learning (Experiment 2) were investigated in four inbred strains of mice (BALB/c, C3H/He, C57BL/6 and DBA/2J). In Experiment 1, mice received a one-second inescapable shock, which ranged in intensity from .01 to .4 mA (12 levels). Response types to shock were observed, and locomotion increased in all strains as the shock intensity increased. Additionally, the C3H/He strain also increased jumping response at shocks greater than .2 mA. In Experiment 2, the four strains were trained for shuttle avoidance with three shock intensities (.06, .16 and .4 mA). Two responses, locomotion into the adjacent compartment (L-typed) and rearing or jumping (R-typed), were equally effective in terminating the shock. While all strains learned the task at all the levels of shock intensities, the performance of the BALB/c strain declined as the intensity decreased. BALB/c and C57BL/6 strains avoided the shocks mainly by L-typed response across all the intensities. Likewise, the DBA/2J strain predominately displayed L-typed responses, but some R-typed responses did occur. The C3H/He strain, on the other hand, largely avoided the shocks by the R-typed response, especially in the .06 and .4 mA conditions.

[Sequential structure of behavior and its modification in inbred strains of mice in a passive avoidance situation].

Behavior in a shuttle-box of four inbred strains of mice (BALB, C3H, C57BL and DBA) was observed by time-sampling method with 11 behavioral items in order to determine how their dyadic sequential structure of the items would be modified under a passive avoidance procedure. Mice in the safe-compartment could enter freely into the shock-compartment, the grid of which was electrified during Days 1-5. A common structure mainly composed of six items was found in every strain on Day 0 when the grid floor was not electrified. The structure was modified on Days 1-5. Entries into the shock-compartment were greatly reduced in all strains as the shock-experience increased, indicating the establishment of passive-avoidance. However, the way they avoided shocks was quite different between BALB and the other strains. In the latter the transitions between 'locomotion' and 'peeping' and between 'sniffing' and 'peeping' did not extinguish, while in BALB these transitions extinguished after Day 1, which means they avoided even 'peeping' into the shock-compartment. It was concluded that the passive avoidance strategy may be different among inbred strains.

[An active part of female mice in sexual interaction].

Proceptive behavior, according to Beach (1976), maintains and accelerates sexual interactions toward the end goal. Such distinctive type of proceptive behavior as darting or hopping of female rats is not apparent in mice. Nevertheless, it seems more reasonable that females may take an active part as much as their male partners also in this species. Twenty sexually experienced females in estrus were paired with 20 naive males of the same strain (ICR/JCL) and the pairs were observed for 6 hours. Eleven males of the 20 pairs successfully ejaculated. Females exhibited lordosis more frequently when they actively approached the male partners than when they were approached and mounted by males. This indicated that the rate of mating success was higher in the case of female's approach. Female's approaching behavior thus possibly plays a role as a proceptive behavior in mice.

A system for study of coronavirus mRNA synthesis: a regulated, expressed subgenomic defective interfering RNA results from intergenic site insertion.

A system that exploits defective interfering (DI) RNAs of mouse hepatitis virus (MHV) for deciphering the mechanisms of coronavirus mRNA transcription was developed. A complete cDNA clone of MHV DI RNA containing an inserted intergenic region, derived from the area of genomic RNA between genes 6 and 7, was constructed. After transfection of the in vitro-synthesized DI RNA into MHV-infected cells, replication of genomic DI RNA as well as transcription of the subgenomic DI RNA was observed. S1 nuclease protection experiments, sequence analysis, and Northern (RNA) blotting analysis revealed that the subgenomic DI RNA contained the leader sequence at its 5' end and that the body of the subgenomic DI RNA started from the inserted intergenic sequence. Two subgenomic DI RNAs were synthesized after inserting two intergenic sites into the MHV DI RNA. Metabolic labeling of virus-specific protein in DI RNA replicating cells demonstrated that a protein was translated from the subgenomic DI RNA, which can therefore be considered a functional mRNA. Transfection study of gel-purified genomic DI RNA and subgenomic DI RNA revealed that the introduction of the genomic DI RNA, but not subgenomic DI RNA, into MHV-infected cells was required for synthesis of the subgenomic DI RNA. A series of deletion mutations in the intergenic site demonstrated that the sequence flanking the consensus sequence of UCUAAAC affected the efficiency of subgenomic DI RNA transcription and that the consensus sequence was necessary but not sufficient for the synthesis of the subgenomic DI RNA.