Screening Reveals Sterol Derivatives with Pro-Differentiation, Pro-Survival, or Potent Cytotoxic Effects on Oligodendrocyte Progenitor Cells.

Inducing the formation of new oligodendrocytes from oligodendrocyte progenitor cells (OPCs) represents a potential approach to repairing the loss of myelin observed in multiple sclerosis and other diseases. Recently, we demonstrated that accumulation of specific cholesterol precursors, 8,9-unsaturated sterols, is a dominant mechanism by which dozens of small molecules enhance oligodendrocyte formation. Here, we evaluated a library of 56 sterols and steroids to evaluate whether other classes of bioactive sterol derivatives may also influence mouse oligodendrocyte precursor cell (OPC) differentiation or survival. From this library, we identified U-73343 as a potent enhancer of oligodendrocyte formation that induces 8,9-unsaturated sterol accumulation by inhibition of the cholesterol biosynthesis enzyme sterol 14-reductase. In contrast, we found that mouse OPCs are remarkably vulnerable to treatment with the glycosterol OSW-1, an oxysterol-binding protein (OSBP) modulator that induces Golgi stress and OPC death in the low picomolar range. A subsequent small-molecule suppressor screen identified mTOR signaling as a key effector pathway mediating OSW-1's cytotoxic effects in mouse OPCs. Finally, evaluation of a panel of ER and Golgi stress-inducing small molecules revealed that mouse OPCs are highly sensitive to these perturbations, more so than closely related neural progenitor cells. Together, these studies highlight the wide-ranging influence of sterols and steroids on OPC cell fate, with 8,9-unsaturated sterols positively enhancing differentiation to oligodendrocytes and OSW-1 able to induce lethal Golgi stress with remarkable potency.

Factors associated with advanced-stage Merkel cell carcinoma at initial diagnosis and the use of radiation therapy: Results from the National Cancer Database.

BACKGROUND: The stage of disease at initial diagnosis and the use of radiation therapy (RT) are important determinants of survival in patients with Merkel cell carcinoma (MCC). OBJECTIVE: To define factors that are associated with advanced-stage MCC at the time of initial diagnosis and the use of RT. METHODS: Cross-sectional, retrospective analysis of patients with MCC registered in the National Cancer Database during the period from 2004 to 2013. RESULTS: A total of 11,917 patients were identified; 3152 and 4586 patients were excluded from the staging and RT analyses, respectively, because of lack of available data. African American ethnicity (odds ratio [OR], 1.49; 95% confidence interval [CI], 1.06-2.10; P = .023), lack of medical insurance (OR, 2.15; 95% CI, 1.40-3.30; P < .001), Charlson-Deyo comorbidity score of at least 1 (OR, 1.21; 95% CI, 1.09-1.34; P < .001), residence more than 26 miles from a treatment facility (OR, 1.18; 95% CI, 1.03-1.35; P = .015), tumor located on the lower limb/hip (OR, 1.59; 95% CI, 1.42-1.78; P < .001) or trunk (OR, 2.05; 95% CI, 1.81-2.33; P < .001), and poorly (OR, 2.57; 95% CI, 1.13-5.82; P = .024) or undifferentiated (OR, 3.11; 95% CI, 1.36-7.15; P = .007) tumor histology predicted advanced-stage MCC at the time of initial diagnosis. The use of RT was associated with Native American ethnicity (OR, 5.04; 95% CI, 1.10-22.99; P = .037), tumor size between 1.5 and 2.7 cm (OR, 1.27; 95% CI, 1.10-1.47; P = .001), electing not to have surgery (OR, 2.77; 95% CI, 1.90-4.03; P < .001), positive postsurgical margins (OR, 1.39; 95% CI, 1.18-1.63; P < .001), and receiving treatment at a comprehensive cancer program (OR, 1.25; 95% CI, 1.03-1.50; P = .020). LIMITATIONS: Retrospective design limits generalizability of the results, and precise details of RT regimens utilized were not available. CONCLUSIONS: A number of factors are associated with advanced-stage MCC at initial diagnosis and the use of RT. Health care models should account for these factors, and efforts should be directed toward improving those that are modifiable.

Quantitative trait locus analysis of neointimal formation in an intercross between C57BL/6 and C3H/HeJ apolipoprotein E-deficient mice.

BACKGROUND: Inbred mouse strains C57BL/6J (B6) and C3H/HeJ (C3H) exhibit marked differences in neointimal formation after arterial injury when deficient in apolipoprotein E (apoE(-/-)) and fed a Western diet. Quantitative trait locus (QTL) analysis was performed on an intercross between B6.apoE(-/-) and C3H.apoE(-/-) mice to determine genetic factors contributing to the phenotype. METHODS AND RESULTS: Female B6.apoE(-/-) mice were crossed with male C3H.apoE(-/-) mice to generate F(1)s, which were intercrossed to generate 204 male F(2) progeny. At 10 weeks of age, F(2)s underwent endothelium denudation injury to the left common carotid artery. Mice were fed a Western diet for 1 week before and 4 weeks after injury and analyzed for neointimal lesion size, plasma lipid and MCP-1 levels. One significant QTL, named Nih1 (61cM, LOD score: 5.02), on chromosome 12 and a suggestive locus on chromosome 13 (35cM, LOD: 2.67) were identified to influence lesion size. One significant QTL on distal chromosome 1 accounted for major variations in plasma non-HDL cholesterol and triglyceride levels. Four suggestive QTLs on chromosomes 1, 2, and 3 were detected for circulating MCP-1 levels. No correlations were observed between neointimal lesion size and plasma lipid levels or between lesion size and plasma MCP-1 levels. CONCLUSIONS: Neointimal formation is controlled by genetic factors independent of those affecting plasma lipid levels and circulating MCP-1 levels in the B6 and C3H mouse model.

Randomized comparison of oral valacyclovir and intravenous ganciclovir for prevention of cytomegalovirus disease after allogeneic bone marrow transplantation.

In this multicenter, randomized study, cytomegalovirus (CMV)-seropositive patients who received an allogeneic bone marrow transplant were provided high-dose intravenous acyclovir (500 mg/m(2) q8h) from the day of transplantation until engraftment. The patients were then randomly assigned to receive either oral valacyclovir, 2 g q.i.d. (n=83), or intravenous ganciclovir, 5 mg/kg q12h for 1 week, then 6 mg/kg once daily for 5 days per week (n=85), until day 100 after transplantation. CMV infection occurred in 12% of the patients who received valacyclovir and in 19% of the patients who received ganciclovir (hazard ratio [HR], 1.042; 95% confidence interval [CI], 0.391-2.778; P=.934). CMV disease developed in only 2 patients who received valacyclovir and in 1 patient who received ganciclovir (HR, 1.943; 95% CI, 0.176-21.44; P=.588). Oral valacyclovir can be an effective alternative to intravenous ganciclovir for prophylaxis of CMV disease after bone marrow transplantation.

Tea tree oil reduces the swelling associated with the efferent phase of a contact hypersensitivity response.

OBJECTIVE: To examine the anti-inflammatory activities of tea tree oil (TTO) in vivo. METHODS: Mice were sensitized to a chemical hapten, trinitrochlorobenzene, on their ventral skin and 7 days later challenged (or re-exposed) on their dorsal skin with the same hapten. RESULTS: TTO applied 30 min before or up to 7 h after to the same dorsal site as hapten challenge caused a significant reduction in skin swelling after 24 h. TTO reduced oedema but not the influx of inflammatory cells. This finding was supported by the inability of TTO to suppress TNFalpha-induced E-selectin expression by human umbilical vein endothelial cells. TTO did not suppress irritant- or ultraviolet B-induced oedema. CONCLUSION: Topical TTO, specifically the TTO components, terpinen-4-ol and alpha-terpineol can regulate the oedema associated with the efferent phase of a contact hypersensitivity response.

Localization of the melatonin-related receptor in the rodent brain and peripheral tissues.

Previous studies have provided a limited examination of the expression of the orphan melatonin-related receptor in the pituitary and hypothalamus of human and sheep and retinal tissue in the sheep. The present study reports evidence of conservation of expression in regions of the hypothalamus (dorsal medial hypothalamus, lateral hypothalamus, arcuate nucleus), the epithelial layer lining the third ventricle and the paraventricular thalamic nucleus of the mouse, rat and hamster. An extensive and detailed analysis of melatonin-related receptor mRNA expression in the mouse central nervous system and peripheral tissues is presented. Mapping the distribution throughout the entire mouse brain has revealed new sites of expression in a number of brain nuclei, including preoptic areas, parabrachial nuclei and widespread distribution in the olfactory bulb. Reverse transcriptase-polymerase chain reaction was performed with RNA isolated from peripheral tissues revealing expression of the melatonin-related receptor mRNA in the mouse kidney, adrenal gland, intestine, stomach, heart, lung, skin, testis and ovary. These results suggest a conserved function in neuroendocrine regulation and a potential role in coordinating physiological responses in the central nervous system and peripheral tissues.

Living with rheumatoid arthritis: the role of daily spirituality and daily religious and spiritual coping.

The objective of this preliminary study was to evaluate more fully the role of daily spiritual experiences and daily religious/spiritual coping in the experience of individuals with pain due to rheumatoid arthritis (RA). Thirty-five individuals with RA were asked to keep a structured daily diary for 30 consecutive days. The diary included standardized measures designed to assess spiritual experiences, religious and spiritual pain coping, salience of religion in coping, religious/spiritual coping efficacy, pain, mood, and perceived social support. The participants in this study reported having spiritual experiences, such as feeling touched by the beauty of creation or feeling a desire to be closer or in union with God, on a relatively frequent basis. These participants also reported using positive religious and spiritual coping strategies much more frequently than negative religious and spiritual coping strategies. Although most of the variance in these measures was due to differences between persons, each measure also displayed a significant variability in scores from day to day. Indeed, there was just as much (or more) variability in these measures over time as there was variability in pain. Individuals who reported frequent daily spiritual experiences had higher levels of positive mood, lower levels of daily negative mood, and higher levels of each of the social support domains. Individuals who reported that religion was very salient in their coping with pain reported much higher levels of instrumental, emotional, arthritis-related, and general social support. Coping efficacy was significantly related to pain, mood, and social support in that on days that participants rated their ability to control pain and decrease pain using spiritual/religious coping methods as high, they were much less likely to have joint pain and negative mood and much more likely to have positive mood and higher levels of general social support. Taken together, these results suggest that daily spiritual experiences and daily religious/spiritual coping variables are important in understanding the experience of persons who have RA. They also suggest that newly developed daily diary methods may provide a useful methodology for studying religious and spiritual dimensions of living with arthritis.

The ovine melatonin-related receptor: cloning and preliminary distribution and binding studies.

A melatonin-related receptor was cloned from an ovine genomic library. The sequenced gene has a similar structure to that of the melatonin receptor gene family and consists of two exons separated by an intron of approximately 3 kb. Exon 1 and exon 2 of the ovine melatonin-related receptor encode a protein of 575 amino acids which is 73.8% homologous to the human melatonin-related receptor and shows 40.9% homology with the ovine Mel1a melatonin receptor. COS-7 cells transiently expressing ovine melatonin-related receptors did not bind 2-[125]iodomelatonin or 3H-melatonin. Reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization studies revealed expression of the ovine melatonin-related receptor in the hypothalamus, pituitary, retina and retinal pigment epithelium. Furthermore, expression of the ovine melatonin-related receptor is shown to be coincident with Mel1a and 2-[125I]iodomelatonin binding in the pituitary and serotonin N-acetyl transferase (arylalkylamine N-acetyl transferase, AANAT) expression in the retina. Expression patterns and similarity with the melatonin receptor gene family suggest a role for this novel G protein-coupled receptor in control and regulation of endocrine function and retinal physiology.

Sequence of a novel plant ras-related cDNA from Pisum sativum.

A clone isolated from a purple podded pea (Pisum sativum L.) cDNA library was shown to contain the complete coding sequence of a polypeptide with considerable homology to various members of the ras superfamily. The ras superfamily are a group of monomeric GTP-binding proteins of 21-25 kDa found in eukaryotic cells. Conserved sequences in the isolated clone include the GTP-binding site, GDP/GTP hydrolysis domain and C-terminal Cys residues involved in membrane attachment. Comparisons of the predicted amino acid sequence with those of other ras proteins show significantly higher homologies (ca. 70%) to two mammalian gene products, those of the BRL-ras oncogene, and the canine rab7 gene, than to any of the plant ras gene products so far identified (< 40% homology). The high percentage of amino acid identity suggests that this cDNA may be the product of a gene, designated Psa-rab, which is the plant counterpart of rab7. Rab/ypt proteins are a subfamily of the ras superfamily thought to be involved in intracellular transport from the endoplasmic reticulum to the Golgi apparatus and in vesicular transport. Northern blot hybridisation analysis of total RNA from green and purple podded pea revealed a mRNA species of approximately the same size as the isolated cDNAs.

Product inhibition of the actomyosin subfragment-1 ATPase in skeletal, cardiac, and smooth muscle.

We studied product inhibition of the actin-activated ATPase of myosin subfragment-1 (S-1) from the three types of muscle tissue: skeletal, cardiac, and smooth. Increasing levels of [MgADP] in the 0-1-mM range caused significant inhibition of the actin-activated MgATPase activity of cardiac and gizzard but not skeletal muscle S-1. When total nucleotide concentration ([ATP] + [ADP]) was kept constant at 1 mM, ATPase activity was inhibited by 50% at an ADP/ATP ratio of 6:1 for cardiac S-1 and 3:1 for gizzard S-1. For skeletal S-1, however, even a 19:1 ratio did not cause 50% inhibition of ATPase activity. The observed effect was not due to changes in pH or inorganic phosphate concentration, nor could it be explained by substrate (ATP) depletion. In the absence of actin, ADP had little or no inhibitory effect on the ATPase activity of S-1, and these observations imply that ADP is competing directly for the ATP binding site of the actin-S1 complexes of cardiac and smooth muscle S-1. ADP has previously been shown to be a weak competitive inhibitor of the ATPase activity in skeletal muscle. The current data imply that ADP is a very effective competitive inhibitor for the actin-activated ATPase activity of cardiac and gizzard S-1 and, therefore, that ADP may be a physiologically important modulator of contractile activity in cardiac and smooth muscle.

Jaundice: clinical practice in 88,000 liveborn infants.

We reviewed jaundiced infants born between 1971 and 1989. Jaundice was diagnosed in infants whose serum bilirubin level was found to be 154 umol/l or greater. Of 88,137 livebirths, 10,944 (12.4%) were jaundiced. The most common aetiological factor was prematurity (20.3%), followed by ABO erythroblastosis (5.5%), sepsis (1.8%), Rh erythroblastosis (1.8%), bruising (1.3%), multifactorial (1.0%) and glucose-6-phosphate dehydrogenase deficiency (0.5%). In the remainder (67.8%) no cause was found or inadequate investigations were performed to determine a cause. During the period under review there was a significant increase (r = 0.91) in the proportion of newborn infants with jaundice of prematurity, in those not investigated (r = 0.92) and a decrease in the proportion with bruising (r = -0.90) as the cause. Phototherapy was used on 4,126 (37.7%) infants and exchange transfusion performed on 248 (2.3%). Causes of jaundice in infants requiring exchange transfusion were Rh erythroblastosis (108, 43.6%), ABO erythroblastosis (58, 23.4%), jaundice of prematurity (44, 17.7%) and a variety of causes in the remaining 38 (15.3%). Death occurred in 164 (1.5%) infants. In only 7 (4.3%), however, was the death possibly related to hyperbilirubinaemia or its treatment (Rh erythroblastosis (4), necrotizing enterocolitis following exchange transfusion (2) and pulmonary haemorrhage following exchange transfusion (1)). Phototherapy proved safe with no deaths attributable to its use.

Preparation of lipid emulsions by pressure extrusion.

Lipid emulsions consisting of a surface monolayer of phospholipid enclosing a core of neutral lipids have been prepared by repeated extrusion through polycarbonate filters of defined pore size. Particle size, as measured by photon correlation spectroscopy, decreases on successive passes through a 100 nm filter, reaching a near constant value (130-150 nm) after 4 passes. A corresponding decrease in the standard deviation of the particle size distribution occurs during this process. The recovery of lipids, especially of cholesterol and cholesterol ester, is improved if the emulsion is sonicated before extrusion through filters. [31P]-NMR and fluorescence techniques are used to confirm that the resulting structures are emulsions rather than lipid bilayers.

Outcomes following the use of rational guidelines in the management of jaundiced newborn infants.

In 1975, rational guidelines for management of the jaundiced newborn infant were introduced to the Mercy Maternity Hospital, Melbourne. The guidelines were produced as four charts, each chart covering a particular birthweight range. The charts have been used widely. The effect of introduction of the charts has been examined by comparing the years 1971-74 with 1975-77. An overall decrease occurred in the use of phototherapy, without an increase in the use of exchange transfusion or in those with a serum bilirubin level above 339 mumol/L. The difference was not explained by an alteration in the spectrum of causes of jaundice. Long-term follow-up of jaundiced infants managed according to these guidelines revealed a satisfactory outcome despite a significant reduction in active treatment. The associated avoidance of potential side effects of treatment of the jaundiced newborn infant warrants consideration of the use of these charts by other neonatal units.

Cephalhaematoma: a 10-year review.

The incidence of cephalhaematoma at the Mercy Maternity Hospital over a 10-year period was 2.5%; of the 1,030 infants 68.4% were born to primiparas, 65.6% were males, the majority (91.1%) were between 37 and 42 weeks' gestation and 3,000 and 4,000 g birth-weight (71.6%). Forceps delivery and vacuum extraction were associated with increased incidences of cephalhaematoma (5.1% and 22.9% respectively), and the incidence was slightly increased (3.8%) when a scalp electrode had been applied. Hyperbilirubinaemia was more prevalent (12.9%) in infants with a cephalhaematoma as was exchange transfusion (0.8%) and the need for phototherapy (4.9%).

Jaundice: a 10 year review of 41,000 live born infants.

A review is presented of jaundiced newborn infants during the 10-year period to 1980. Included are those whose serum bilirubin level was 154 mumol/l or more. Of 41,057 live births, 4,406 (10.7%) infants had hyperbilirubinaemia. The most common (19.9;%) aetiological factor was prematurity, followed by ABO erythroblastosis 7.1%; sepsis 3.4%; Rhesus erythroblastosis 2.7%; bruising 2.2%; multifactorial 1.0% and glucose-6-phosphate dehydrogenase deficiency 0.5%. Treatment was not undertaken in 2,855 (64.7%) infants, but 1,419 (32.2%) received phototherapy alone, 122 (2.7%) infants received both exchange transfusion and phototherapy and 10 (0.2%) infants received exchange transfusion alone. Of the infants requiring exchange transfusion 50.0% had Rhesus erythroblastosis, 28.0% ABO erythroblastosis, 10.6% jaundice of prematurity and the remainder were due to a variety of causes. Sixty-three (1.4%) infants died, with two deaths being related to the hyperbilirubinaemia, as their death was due to necrotizing enterocolitis following exchange transfusion. Phototherapy proved safe with no deaths directly attributable to its use.

Antifertility activity of N-protected glycine activated esters.

A series of N-protected glycine activated esters was found to have antifertility activity in mice when administered intravaginally. The N-carbobenzoxyglycine vinyl ester and N-carbobenzoxyglycine 1,2-dibromoethyl ester analogs possessed the best activity in inhibiting pregnancy but were much less active when administered intraperitoneally. The acrosin enzymatic activity of sperm also was inhibited by these N-protected glycine activated esters, as measured by N-alpha-benzoyl-L-arginine ethyl ester and azocasein hydrolysis. The ability to inhibit N-alpha-benzoyl-L-arginine ethyl ester hydrolysis, a trypsin-like activity, appeared to have a positive correlation with antifertility activity when the agents were administered intravaginally.

PIP: A series of glycine esters were tested for antifertility activity in mice. N-protected glycine, when administered intravaginally had antifertility activity in mice both in vitro and in vivo. Greatest antifertility activity was found with 2 glycine esters, N-carbobenzoxyglycine vinyl ester, and N-carbobenzoxyglycine 1,2-dibromoethyl ester analogs; 0% pregnancy occurred when administered intravaginally. Their inhibiting activity was much less when administered intraperitoneally. In vitro, the acrosin enzymatic activity of sperm was inhibited by these N-protected glycine-activated esters, as determined by measuring N-alpha-benzoyl-L-arginine ethyl ester and azocasein hydrolysis. The ability to inhibit the arginine ethyl ester hydrolysis, which is a trypsin-like activity, seemed to be the positive correlate with antifertility activity when agents were administered intravaginally.