A single-centre chart review exploring the adjusted association between breast cancer phenotype and prognosis.

PURPOSE AND METHODS: Using a retrospective chart review, we investigated the differences in survival and prognostic factors between patients with triple-negative breast cancer (tnbc) and those with non-tnbc. The review included 1018 breast cancer patients who were diagnosed between 2000 and 2005 in Essex, Kent, and Lambton counties in Ontario, Canada. RESULTS: Our findings indicate that, although the unadjusted results suggested that patients with tnbc were more likely than patients with non-tnbc to die [hazard ratio (hr): 2.29; 95% confidence interval (ci): 1.33 to 2.93], an adjusted survival analysis revealed no significant difference in overall survival between the groups (hr: 1.22; 95% ci: 0.63 to 2.39). The significant predictors of survival in the adjusted analysis were age, stage of cancer, and size of cancer. CONCLUSIONS: Our findings support those of earlier reports, which suggest that presenting tumour size is the most important prognostic factor in tnbc. Investigations into unique screening methods to identify these tumours at an earlier stage and to prevent advanced-stage cancer in this patient subpopulation are necessary.

Aberrant regulation of endogenous ouabain-like factor in bipolar subjects.

Ill phases of bipolar illness are associated with abnormalities in ion regulation and intracellular ion concentrations. Previously, it has been reported that mania is characterised by lower circulating levels of ion regulating endogenous cardenolides, and that bipolar subjects lack the normal seasonal variation of these factors. Since endogenous cardenolides are elaborated in settings of extensive physical activity, euthymic bipolar and psychiatrically normal control subjects were asked to exercise to exhaustion. Plasma concentrations of endogenous cardenolides were measured at baseline, 60 min, peak exercise and post-recovery. Ouabain-like immunoreactive factor (OLF) was lower at baseline (0.005+/-S.D. 0.01 ng/mL in bipolar vs. 0.072+/-0.06 ng/mL in normal control subjects, P=0.019), lower at 60 min (0.007+/-S.D. 0.02 ng/mL in bipolar vs. 0.075+/-0.06 ng/mL in normal control subjects, P=0.029), and tended to be lower at peak exercise (0.009+/-S.D. 0.02 ng/mL in bipolar vs. 0.131+/-0.21 ng/mL in normal control subjects, P=0.15) in bipolar subjects compared to non-psychiatric controls. Other endogenous cardenolides did not vary significantly. The endogenous cardenolide, OLF, may be aberrantly controlled in bipolar illness.

Efficacy of olanzapine and haloperidol in an animal model of mania.

PURPOSE: Intracerebroventricular (ICV) administration of ouabain, a potent sodium pump inhibitor, has been used to model mania. Antipsychotic agents have demonstrated efficacy in the management of acute mania. This study was undertaken to determine the prophylactic efficacy of olanzapine and haloperidol in the ouabain mania model. METHODS: Male Sprague-Dawley rats (4-8/group) were treated with two haloperidol decanoate intramuscular shots one week apart (21 mg/kg) or twice daily olanzapine intraperitoneal injections at low dose (1 mg/kg/day) or high dose (6 mg/kg/day) for 7 days prior to ICV administration of ouabain. Open field locomotion was quantified at baseline and after ouabain administration. RESULTS: Ouabain caused a significant increase in open field locomotion (253.7+/-SEM 55.12 vs control 53.1+/-12.13 squares traversed in 30 min in the olanzapine experiments, P<0.05; and 236.5+/-41.42 vs 129.3+/-38.23, P<0.05 in the haloperidol experiments). Olanzapine alone at low dose (102.2+/-37.7) or high dose (151.2+/-49.2) did not alter open field activity. Low dose olanzapine (176.6+/-73.27) but not high dose (307.5+/-167.32) caused a modest reduction of the ouabain effect. Haloperidol alone significantly reduced motoric activity compared to control (55.6+/-18.0, P<0.05), and prevented ouabain-induced hyperactivity (60.3+/-33.1, P<0.05). CONCLUSION: Haloperidol, but not olanzapine, demonstrated efficacy in this mania model, but methodological details may have reduced the effect of olanzapine.

Glial cell number and neuron/glial cell ratios in postmortem brains of bipolar individuals.

BACKGROUND: There are several lines of evidence implicating a glial abnormality in the pathophysiology of bipolar disorder. Previous studies have reported a wide range of abnormalities but with little consistency between their findings. METHODS: Sixty frozen postmortem temporal cortical brain samples from normal, schizophrenic, bipolar, and depressed subjects were obtained from the Stanley Neuropathology Consortium. Nissl stain was utilized to visualize cellular structures. The sections were analyzed for neuron and glial content using the computer programs SPOT Advanced and Metamorph. Three representative fields from each subject were counted and the average numbers obtained. RESULTS: There was no critical difference in glia number across the four diagnostic groups. Compared with normal controls, area occupied by glia was reduced in bipolar subjects (P = 0.018), and the ratio of glial area to neuronal area was reduced (P = 0.028). DISCUSSION: The percent glial space was substantially lower in bipolar disorder subjects compared to normal controls, suggesting that the glia may be smaller in bipolar subjects. The size reduction may be reflective of glial dysfunction. LIMITATIONS: Postmortem interval of 29.4 +/- S.D. 13.4 h may have affected cellular structure and reduced the quality of the staining.

Testing for viral penetration of non-latex surgical and examination gloves: a comparison of three methods.

Currently, there are no international standards based on microbiological methodology for testing the ability of medical examination or surgical gloves to prevent the passage of viruses. Three protocols for the direct examination of the viral barrier properties of non-latex gloves were compared with 1080 gloves (270 gloves from each of two surgical brands and two medical examination brands). In two of the methods, gloves were filled with and suspended in a nutrient broth solution, and bacteriophage phiX174 was placed either inside or outside the glove, while the entire test vessel was agitated. Gloves tested using the third method were filled with a suspension of bacteriophage and allowed to rest in a vessel containing nutrient broth. Gloves were tested directly from the manufacturer's packaging, or after being punctured intentionally or subjected to a stress protocol. The passage of bacteriophage was detected with plaque assays. Significant differences in failure rates between glove brands were apparent only among gloves that had been subjected to the stress protocol. Overall, the two methods in which bacteriophage were placed inside the gloves provided more sensitivity than the method in which bacteriophage was spiked into broth outside the gloves. Thus the placement of bacteriophage inside test gloves (or the use of pressure across the glove barrier during testing), and the use of a standardised stress protocol, will improve significantly the ability of a glove test protocol to determine the relative quality of the barrier offered by medical examination and surgical gloves. Further research is needed to provide test methods that can incorporate reproducibly both the use of bacteriophage and simulated glove use in an industrial quality control setting.

Intracerebroventricular administration of ouabain as a model of mania in rats.

BACKGROUND: Human bipolar illness is characterized by mood state- and diagnosis-associated abnormalities of cellular cation distribution and transport. These include reduced sodium pump activity and expression and increased intracellular sodium. If these alterations are related to the pathophysiology of the disease, rather than secondary or ancillary abnormalities, then one would expect that modeling of these changes in vivo would produce lithium-preventable behavioral abnormalities. METHODS: Ouabain, a potent inhibitor of the sodium pump, was administered intracerebroventricularly to male rats previously fed lithium-containing food or plain rat chow. Locomotion was then quantified in an open field. RESULTS: Ouabain increased locomotion 300% over baseline. Lithium pretreatment prevented the ouabain-induced hyperlocomotion response. CONCLUSION: Inhibition of central nervous system sodium pump with ouabain produces a plausible animal model of mania. This model may be useful for preclinical screening of potential mood stabilizers.

Effect of ethacrynic acid on sodium pump alpha isoforms in SH-SY5Y cells.

BACKGROUND: Ethacrynic acid (ECA), a diuretic that has several cellular actions, increases expression of the sodium and potassium-activated adenosine triphosphatase (Na, K-ATPase or Na pump) in normal lymphocytes, but not in lymphocytes of bipolar patients. While this has been proposed to be important in the pathophysiology of bipolar illness, the response of neural tissues to ECA is unknown. METHODS: Human neuroblastoma SH-SY5Y cells differentiated with 10-microM retinoic acid were treated with various ECA concentrations for 3 days, and changes in Na-pump alpha-isoform expression were quantified with densitometric analysis of Western bands. RESULTS: Expression of alpha1 and alpha3 Na pump isoforms significantly increased with 10-5 M ECA. Cells treated with 10-6 or 10-7 M ECA showed no change in Na-pump expression, while cells treated with 10-4 M ECA died. The alpha2 isoform could not be detected in differentiated SH-SY5Y cells. CONCLUSIONS: The effect of ECA on alpha1-isoform in neural tissue is similar to that observed in lymphocytes. As alpha3 isoform is not expressed in lymphocytes, however, we conclude that lymphocytes are an incomplete model of neural tissue.

Human adrenal cells in culture produce both ouabain-like and dihydroouabain-like factors.

BACKGROUND: Ouabain-like factor (OLF) and its newly discovered reduced species, dihydroouabain-like factor (Dh-OLF), are mammalian cardenolides whose structural and functional characteristics are similar to the plant-derived compounds ouabain and dihydroouabain. These endogenous compounds are believed to be produced by the adrenals and to constitute part of an hormonal axis that may regulate the catalytic activity of the alpha-subunit of Na(+),K(+)-ATPase. We developed antibodies sufficiently specific to distinguish between OLF and Dh-OLF, and in this study demonstrate the selective secretion of OLF and Dh-OLF from human H295R-1 adrenocortical cells in culture. METHODS: We used reversed-phase HPLC, inhibition of Na(+),K(+)-ATPase catalytic activity, and two enzyme immunoassays developed with antibodies specific to ouabain and dihydroouabain to purify and characterize the secretion of these two compounds by human adrenal cells in culture. Purified antisera had high titers (1 x 10(6) for ouabain and 8 x10(5) for dihydroouabain) and were specific to their corresponding antigens. RESULTS: Human H295R-1 cells grown in serum-free medium secreted 0.18 +/- 0.03 pmol of OLF and 0.39 +/- 0.04 pmol of Dh-OLF per 10(6) cells in 24 h. Both OLF and Dh-OLF inhibited the ouabain-sensitive catalytic activity of the sodium pump (0.03 micro mol/L OLF inhibited 29% of the catalytic activity; 0.07 micro mol/L Dh-OLF inhibited 17%). Stimulation of the cell culture by dibutryl cAMP increased the secretion of Dh-OLF 50% over control (unstimulated), whereas the secretion of OLF did not increase significantly. CONCLUSIONS: OLF and Dh-OLF are secreted by human adrenal cells, and antibodies specific to these two compounds can be developed, using the plant-derived counterparts as antigens. The secretion of Dh-OLF is responsive to a cAMP-dependent stimulation mechanism, whereas OLF is not. Our data suggest that either the secretory or biosynthetic pathways for production of these two compounds by human adrenal cells may have different control mechanisms or that they may be linked via a precursor-product relationship.

Evaluation of neuroprotection by lithium and valproic acid against ouabain-induced cell damage.

BACKGROUND: The pathophysiology of manic-depression may be associated with dysregulation of ion homeostasis. Ouabain is a potent inhibitor of the sodium-potassium adenosine triphosphatase and has been purported to mimic abnormalities seen in acute mania. As manic episodes are believed to be neurotoxic and mood stabilizers have recently been implicated as neuroprotectants, it is of interest to determine if lithium and valproic acid antagonize ouabain-induced neurotoxicity. METHODS: Human neuroblastoma SH-SY5Y cells were differentiated for 12 days then pretreated with lithium or valproic acid for 24 h and then challenged with a 10 microM ouabain insult. Cellular damage was assessed with lactate dehydrogenase (LDH) release, and apoptotic potential of ouabain was evaluated with DNA fragmentation. RESULTS: Ouabain significantly increased LDH release after 72 h of treatment. Lithium pretreatment at 1 mM diminished ouabain-induced LDH release. Valproic acid alone at 100 and 1000 micrograms/mL significantly increased LDH release from the cells. Furthermore, it significantly potentiated ouabain-induced LDH release. DNA fragmentation suggests that ouabain induces apoptosis. CONCLUSIONS: Lithium at the therapeutic level of 1 mM limits the extent of cellular damage caused by 10 microM ouabain in SH-SY5Y cells as measured by LDH release. Valproic acid alone at the therapeutic concentration of 100 micrograms/mL induces LDH release and does not prevent ouabain-induced LDH release.

Lifetime events and posttraumatic stress disorder in 4 postconflict settings.

CONTEXT: Little is known about the impact of trauma in postconflict, low-income countries where people have survived multiple traumatic experiences. OBJECTIVE: To establish the prevalence rates of and risk factors for posttraumatic stress disorder (PTSD) in 4 postconflict, low-income countries. DESIGN, SETTING, AND PARTICIPANTS: Epidemiological survey conducted between 1997 and 1999 among survivors of war or mass violence (aged >/=16 years) who were randomly selected from community populations in Algeria (n = 653), Cambodia (n = 610), Ethiopia (n = 1200), and Gaza (n = 585). MAIN OUTCOME MEASURE: Prevalence rates of PTSD, assessed using the PTSD module of the Composite International Diagnostic Interview version 2.1 and evaluated in relation to traumatic events, assessed using an adapted version of the Life Events and Social History Questionnaire. RESULTS: The prevalence rate of assessed PTSD was 37.4% in Algeria, 28.4% in Cambodia, 15.8% in Ethiopia, and 17.8% in Gaza. Conflict-related trauma after age 12 years was the only risk factor for PTSD that was present in all 4 samples. Torture was a risk factor in all samples except Cambodia. Psychiatric history and current illness were risk factors in Cambodia (adjusted odds ratio [OR], 3.6; 95% confidence interval [CI], 2.3-5.4 and adjusted OR,1.6; 95% CI, 1.0-2.7, respectively) and Ethiopia (adjusted OR, 3.9; 95% CI, 2.0-7.4 and adjusted OR, 1.8; 95% CI, 1.1-2.7, respectively). Poor quality of camp was associated with PTSD in Algeria (adjusted OR, 1.8; 95% CI, 1.3-2.5) and in Gaza (adjusted OR, 1.7; 95% CI, 1.1-2.8). Daily hassles were associated with PTSD in Algeria (adjusted OR, 1.6; 95% CI, 1.1-2.4). Youth domestic stress, death or separation in the family, and alcohol abuse in parents were associated with PTSD in Cambodia (adjusted OR, 1.7; 95% CI, 1.1-2.6; adjusted OR, 1.7; 95% CI, 1.0-2.8; and adjusted OR, 2.2; 95% CI, 1.1-4.4, respectively). CONCLUSIONS: Using the same assessment methods, a wide range of rates of symptoms of PTSD were found among 4 low-income populations who have experienced war, conflict, or mass violence. We identified specific patterns of risk factors per country. Our findings indicate the importance of contextual differences in the study of traumatic stress and human rights violations.

Secretion of a lactone-hydrogenated ouabain-like effector of sodium, potassium-adenosine triphosphatase activity by adrenal cells.

Ouabain-like factor (OLF), a mammalian cardenolide, is a counterpart to plant-derived ouabain and is found in the adrenal, hypothalamus, and blood of several mammalian species. We now report the existence of a mammalian lactone-hydrogenated ouabain-like factor (dihydro-OLF) in secretions from cultured mouse adrenal Y-1 cells. Dihydro-OLF structurally and functionally mimics plant-derived dihydroouabain. We measured both OLF and the newly discovered dihydro-OLF using five independent techniques: immunoreactivity with two specific antisera, one against ouabain and one against dihydroouabain; chromatographic mobility; spectral absorbance characteristics; and concentration-dependent inhibition and phosphorylation of Na,K-adenosine triphosphatase. All measured physical attributes of dihydro-OLF mimic those of plant-derived dihydroouabain, including a spectral shift maxima, 220 nm (OLF) to 196 nm (dihydro-OLF), with appropriately decreased molar absorptivity. Dihydro-OLF (IC50 = 590 nM) is a 10-fold less potent Na+,K+-adenosine triphosphatase inhibitor than its oxidized mammalian counterpart OLF (IC50 = 60 nM), just as dihydroouabain is less potent than ouabain. Dihydro-OLF is also 3-fold more potent than a recently identified isomer of plant-derived dihydroouabain (IC50 = 1,700 nM). Using antiouabain and antidihydroouabain antisera we estimate that 3 x 10(7) mouse adrenal Y-1 cells secreted 1.3 ng OLF and 8.9 ng dihydro-OLF. The relative abundance of dihydro-OLF is consistently greater than that of its oxidized form, OLF, in bovine adrenals (22-fold), human serum (13-fold), and secretions from cultured mouse Y-1 cells (5-fold). The discoveries of OLF, OLF-genin, and now dihydro-OLF constitute an intriguing structural polymorphism probably involved in the synthesis, regulation, and metabolic control of these new hormone-like compounds.

Emotional and behavioural problems and trauma exposure of school-age Palestinian children in Gaza: some preliminary findings.

An international research team was established to design and implement a community-based, cross-section epidemiological survey of 669 school-age Palestinian children and their families living in the Gaza Strip. Several psychometric instruments were utilized for measuring mental health outcomes, including the Ontario Child Health Scale (OCHS) for measuring childhood emotional and behavioural problems, and the Child Post-traumatic Stress Reaction Index (CPTSRI) to measure post-traumatic stress disorder (PTSD). Lifetime trauma exposure was assessed by using the Health Reach Modified War Questionnaire. It was found that children and adolescents from age six to 16 have high prevalence rates of conduct, attention deficit-hyperactivity disorders and PTSD. These rates are higher than those reported internationally on children in non-conflict areas. A significant correlation was found between higher rates of lifetime trauma exposure, by frequency and type, and higher prevalence rates of mental health problems. The various state and non-governmental organizations involved in health care services are urged to use a community-based approach to alleviate the burden of suffering, including addressing issues of social justice for Palestinians.

Two biologically active isomers of dihydroouabain isolated from a commercial preparation.

Ouabain is a plant-derived cardiac glycoside that inhibits the catalytic activity of Na(+),K(+)-ATPase (sodium pump; NKA). Dihydroouabain, a derivative of ouabain with a reduced lactone ring, is commonly used as a sodium pump antagonist. It has been assumed that commercially available dihydroouabain is homogeneous. We now report that preparations of dihydroouabain contain two components each with a different potency for inhibition of sodium pump activity. We used reverse-phase HPLC chromatography, UV spectrophotometry, electrospray ionization-mass spectrometry (ESI-MS), nuclear magnetic resonance (NMR) spectroscopy and two independent bioassays to characterize these compounds. The two dihydroouabain fractions (Dho-A and Dho-B) resolved by 3 min chromatographically, had UV absorbance maxima at 196 nm, and comprised 37% and 63% of the stock dihydroouabain, respectively. The molar potency of each component for inhibition of NKA from porcine cerebral cortex differed by 4. 4-fold (Dho-A, IC(50) = 7.13 +/- 0.8 microM; Dho-B, IC(50) = 1.63 +/- 0.12 microM). The relative potencies were 9% and 40% of those of ouabain, respectively. A similar pattern for phosphorylation of NKA was observed. Mass spectrometry (ESI-MS) and fragmentation patterns are consistent with Dho-A and Dho-B being isomers of identical molecular mass (587 Da) and each with six hydroxyl groups, a deoxyhexose sugar moiety and a lactone ring. Furthermore, NMR spectroscopy revealed structural differences between Dho-A and Dho-B by displaying noticeably different chemical shifts at only two groups of proton resonances assigned to H-21 and H-22. The ESI-MS and NMR results confirm the presence of the isomerism at C20 of the lactone ring. Our results demonstrate the existence of two molecular forms of dihydroouabain, each with a different biological potency. These findings underscore the importance of characterizing the purity of dihydroouabain commercial preparations. It also provides possible molecular models for investigating the metabolism of endogenous ouabain-like factors recently reported in mammals.

Seasonal distribution and hormonal modulation of reptilian T cells.

Using PNA and anti-Thy-1 fluorescent binding assays, T lymphocytes of the lizard, Chalcides ocellatus were phenotypically distinguishable into four subpopulations (PNA+ Thy-1-, PNA+ Thy-1+, PNA- Thy-1+ and PNA-Thy-1-), which seemed to be affected independently by endogenous steroid levels. Indeed, the size of PNA+ thymocytes is maximal and coincides with the low level of circulating cortisol during spring through summer and decreases gradually with the elevation of the cortisol level. On the other hand, as the endogenous testosterone (TS) level begins its physiological rise, lympholysis of Thy-1+ thymic cells begins in spring with gradual increase in size and with the decrease in TS levels. Among splenocytes and bone marrow lymphocytes, seasonal-dependent alterations in the size of both lymphocyte subpopulations seemed to correlate in part with the status of the thymus. Direct support of this observation was derived from subsequent in vitro studies with exogenous hydrocortisone (HC) and testosterone propionate (TP) treatments in spring and autumn. In all incidents, the data were indicative of the selective susceptibility of the PNA+ Thy-1- subpopulation to HC in the thymus and not in the periphery, and the susceptibility of the PNA- Thy-1+ subpopulation to TP in all three lymphoid organs tested. In vivo studies with a purified fraction of thymosin alpha 1 (T alpha 1) suggested that the PNA+ Thy-1- subpopulation in the different organs was the selective target for the action of T alpha 1. Finally, the dual treatment with T alpha 1 in vivo followed by TP or HC in vitro confirmed that TP-sensitivity was confined to the PNA- Thy-1+ and HC to PNA(+)-Thy-1- subpopulations in any of the three lymphoid organs.

Cryptosporidiosis in children in Gaza.

A prospective survey of children admitted with gastro-enteritis to the Nasser Children's Hospital, Gaza revealed that 19% were excreting cryptosporidium, a significantly (p less than 0.001) greater percentage than that (7%) observed in children admitted for other reasons. Detection of cryptosporidium decreased when the change from hot dry to colder wetter weather occurred. Although Salmonella spp were isolated more frequently than cryptosporidium in children with diarrhoea (20% of cases), there was no statistically significant association between excretion of Salmonella spp and gastro-enteritis. A follow-up study of a cohort of the children with cryptosporidiosis indicated that over three-quarters were dehydrated and all were below their expected weight-for-age. There was a statistically significant association between cryptosporidium gastro-enteritis and evidence of respiratory tract infection.

The Dyggve-Melchio-Clausen syndrome.

Two families with Dyggve-Melchior-Clausen syndrome are reported. In the first family, Jews from Morocco, six of 10 siblings are affected. In the second family, a consanguineous marriage of Arabs from Gaza, two of three children are affected. A description of the skeletal changes in patients ranging in age from 4 to 25 years is presented. The radiologic signs of generalized platyspondyly with double humped end plates and the lace-like appearance of thickened iliac crests are pathognomonic and distinctive of the syndrome. The diagnostic features of the disease are compared to those of Morguio's disease, spondyloepiphyseal dysplasia tarda, and spondylometaphyseal dysplasia.