ABSTRACT
BACKGROUND: In high income countries, over the last three decades, the length of hospital stays for people with serious mental illness has reduced drastically. Some argue that this reduction has led to revolving door admissions and worsening mental health outcomes despite apparent cost savings, whilst others suggest longer stays may be more harmful by institutionalising people to hospital care. OBJECTIVES: To determine the clinical and service outcomes of planned short stay admission policies versus a long or standard stay for people with serious mental illnesses. SEARCH STRATEGY: We searched the Cochrane Schizophrenia Group's register of trials (July 2007). SELECTION CRITERIA: We included all randomised trials comparing planned short with long/standard hospital stays for people with serious mental illnesses. DATA COLLECTION AND ANALYSIS: We extracted data independently. For dichotomous data we calculated relative risks (RR) and their 95% confidence intervals (CI) on an intention-to-treat basis based on a fixed effects model. We calculated numbers needed to treat/harm (NNT/NNH) where appropriate. For continuous data, we calculated fixed effects weighted mean differences (WMD). MAIN RESULTS: We included six relevant trials. We found no significant difference in hospital readmissions between planned short stays and standard care at one year (n=651, 4 RCTs, RR 1.26 CI 1.0 to 1.6). Short hospital stay did not confer any benefit in terms of 'loss to follow up compared with standard care (n=453, 3 RCTs, RR 0.87 CI 0.7 to 1.1). There were no significant differences for the outcome of 'leaving hospital prematurely' (n=229, 2 RCTs, RR 0.77 CI 0.3 to 1.8). More post-discharge day care was given to participants in the short stay group (n=247, 1 RCT, RR 4.52 CI 2.7 to 7.5, NNH 3 CI 2 to 6) and people from the short stay groups were more likely to be employed at two years (n=330, 2 RCTs, RR 0.61 CI 0.5 to 0.8, NNT 5 CI 4 to 8). Economic data were few but, once discharged, costs may be more for those allocated to an initial short stay. AUTHORS' CONCLUSIONS: The effects of hospital care and the length of stay is important for mental health policy. We found limited data, although outcomes do suggest that a planned short stay policy does not encourage a 'revolving door' pattern of admission and disjointed care for people with serious mental illness. More large, well-designed and reported trials are justified.
Subject(s)
Length of Stay , Mental Disorders/rehabilitation , Hospitalization/statistics & numerical data , Humans , Institutionalization , Patient Readmission/statistics & numerical data , Randomized Controlled Trials as TopicABSTRACT
The acylethanolamide anandamide (AEA) occurs in a variety of mammalian tissues and, as a result of its action on cannabinoid receptors, exhibits several cannabimimetic activities. Moreover, some of its effects are mediated through interaction with an ion channel-type vanilloid receptor. However, the chemical features of AEA suggest that some of its biological effects could be related to physical interactions with the lipidic part of the membrane. The present work studies the effect of AEA-induced structural modifications of the dipalmitoylphosphatidylcholine (DPPC) bilayer on phospholipase A2 (PLA2) activity, which is strictly dependent on lipid bilayer features. This study, performed by 2-dimethylamino-(6-lauroyl)-naphthalene fluorescence, demonstrates that the effect of AEA on PLA2 activity is concentration-dependent. In fact, at low AEA/DPPC molar ratios (from R = 0.001 to R = 0.04), there is an increase of the enzymatic activity, which is completely inhibited for R = 0.1. X-ray diffraction data indicate that the AEA affects DPPC membrane structural properties in a concentration-dependent manner. Because the biphasic effect of increasing AEA concentrations on PLA2 activity is related to the induced modifications of membrane bilayer structural properties, we suggest that AEA-phospholipid interactions may be important to produce, at least in part, some of the similarly biphasic responses of some physiological activities to increasing concentrations of AEA.
Subject(s)
1,2-Dipalmitoylphosphatidylcholine/chemistry , Arachidonic Acids/chemistry , Arachidonic Acids/pharmacology , Lipid Bilayers/chemistry , Phospholipases A/metabolism , Cannabinoids , Dose-Response Relationship, Drug , Endocannabinoids , Fluorescent Dyes , Phospholipases A2 , Polyunsaturated Alkamides , Spectrometry, Fluorescence , Structure-Activity Relationship , X-Ray DiffractionABSTRACT
Capacitation is a widely investigated process, which induces sperm plasma membrane changes resulting in its increased affinity for the zona pellucida. The fluorescent probe Laurdan, localized only within the plasma membrane of spermatozoa, is particularly useful to evaluate bilayer polarity in this part of the cell. According to a previous study, sperm membranes from oligozoospermic and some normozoospermic subjects (defined according to World Health Organization criteria), are characterized by low polarity (high Laurdan exGP(340)), while the spermatozoa from the remaining normozoospermic men show a larger polarity (low exGP(340)). In this paper, Laurdan was used to study membrane changes occurring during in-vitro capacitation, on sperm membranes from oligozoospermic and normozoospermic subjects. Results indicated that cells with high exGP(340) show a different susceptibility to Ca(2+)-induced capacitation in vitro, as compared with cells with low exGP(340). Palmitylethanolamide, physiologically present in human reproductive tracts, affects the time-course of in-vitro capacitation, increasing the rate of this process only in the cells with a lower membrane polarity.
Subject(s)
Fluorescence , Sperm Capacitation , Cell Membrane/metabolism , Humans , Infertility , Male , Spermatozoa/metabolism , Zona PellucidaABSTRACT
OBJECTIVE: To determine, by a simple fluorescence method, sperm plasma membrane alterations related with changes of lipid bilayer that, together with routine semen analysis, could help to elucidate the causes of the unexplained male infertility problems. DESIGN: Pilot study. SETTING: Andrology laboratory and biochemistry institute, medical school. PATIENT(S): Men whose semen was studied for infertility problems. INTERVENTIONS(S): No therapeutic intervention was performed on patients. MAIN OUTCOME MEASURE(S): Presence of spermatozoa plasma membrane alterations evidenced by evaluation of Laurdan fluorescence Generalized Polarization (GP) and reported as a function of increasing cell concentration, spermatozoa total motility, linear speed, and vitality. RESULT(S): Reporting GP values as a function of increasing sperm cell concentration, it is evident that the samples are distributed in two distinct areas: at >32 x 10(6) cells per milliliter, mean GP value was 0.303 +/- 0.015, whereas for lower sperm cell concentrations, the mean GP was 0.365 +/- 0.026 (P<.001). These data indicate that the spermatozoa plasma membranes are characterized by liquid-crystalline phases with different ordering degree and polarity and that about 50% of samples with normal semen characteristics (> or =20 x 10(6) cells per milliliter) show high GP values. CONCLUSION(S): Laurdan fluorescence can be used as a simple method to evaluate spermatozoa plasma membrane alterations, particularly in a group of infertile men presenting normal semen parameters. In these samples, Laurdan could be used as a simple tool for infertility assessment. In fact, it is known that compositional and physicochemical alterations of bilayer features can be important for the fertilizing ability of spermatozoa because they are necessary for a proper physiological membrane activity.
Subject(s)
2-Naphthylamine/analogs & derivatives , Cell Membrane/ultrastructure , Infertility, Male/physiopathology , Laurates , Spermatozoa/ultrastructure , Adult , Fluorescent Dyes , Humans , Infertility, Male/pathology , Lipid Bilayers , Male , Pilot Projects , Spectrometry, Fluorescence/methods , Sperm Count , Sperm MotilityABSTRACT
Early notions of mania invoked a combination of psychogenic and organic factors, but psychogenic mania has not endured as a concept. We present a 37-year-old woman with an acute manic episode precipitated by the prolonged death of her husband from cancer. To our knowledge this is the first published account of 'maniacal grief' in the absence of pre-existing affective disorder, and with clear causal relations. Mania was clearly induced by the bereavement and we argue that the loss represented more than a non-specific life event. This case supports a re-examination of mania as potentially a 'reactive' or psychogenic phenomenon.
Subject(s)
Bereavement , Bipolar Disorder/psychology , Psychophysiologic Disorders/psychology , Acute Disease , Adult , Bipolar Disorder/diagnosis , Female , Humans , Life Change Events , Psychophysiologic Disorders/diagnosis , Psychotic Disorders/diagnosis , Psychotic Disorders/psychology , Risk FactorsABSTRACT
The functions of N-acylethanolamines, minor constituents of mammalian cells, are poorly understood. It was suggested that NAEs might have some pharmacological actions and might serve as a cytoprotective response, whether mediated by physical interactions with membranes or enzymes or mediated by activation of cannabinoid receptors. Albumins are identified as the major transport proteins in blood plasma for many compounds including fatty acids, hormones, bilirubin, ions, and many drugs. Moreover, albumin has been used as a model protein in many areas, because of its multifunctional binding properties. Bovine (BSA) and human (HSA) serum albumin are similar in sequence and conformation, but differ for the number of tryptophan residues. This difference can be used to monitor unlike protein domains. Our data suggest that NOEA binds with high affinity to both albumins, modifying their conformational features. In both proteins, NOEA molecules are linked with higher affinity to hydrophobic sites near Trp-214 in HSA or Trp-212 in BSA. Moreover, fluorescence data support the hypothesis of the presence of other NOEA binding sites on BSA, likely affecting Trp-134 environment. The presence of similar binding sites is not measurable on HSA, because it lacks of the second Trp residue.
Subject(s)
Ethanolamines/chemistry , Serum Albumin/chemistry , Circular Dichroism , Fluorometry , Humans , Models, Molecular , Protein Conformation , Protein Structure, Tertiary , Serum Albumin, Bovine/chemistry , Tryptophan/chemistryABSTRACT
BACKGROUND: Lengths for hospital stays for people with serious mental illness have reduced drastically over the last 30 years. Some argue that this reduction has led to revolving door admissions and worsening mental health outcomes despite apparent cost savings, whilst others suggest longer stays may be more harmful in the long term by institutionalising people to hospital care. This review attempts to answer which is the answer: whether short or long stays are effective. OBJECTIVES: To determine the effect of planned short stay admission policies versus a long or standard stay for people with serious mental illnesses. SEARCH STRATEGY: Biological Abstracts (1982-1995), Cochrane Schizophrenia Group's Register (December 1998), EMBASE (1980-1998), MEDLINE (1966-1998) and PsycLIT (1974-1995) were searched. Further references were sought from published trials and their authors. SELECTION CRITERIA: All randomised trials of planned short versus long hospital stays for people with serious mental illness (however defined). DATA COLLECTION AND ANALYSIS: Trials were reliably identified and data extracted. Analysis was on an intention-to-treat basis. People who dropped out or lost to follow-up were assumed to have no improvement. Peto odds ratios (OR) and 95% confidence intervals were calculated. MAIN RESULTS: Five randomised controlled trials were included. For those receiving planned short stays, data suggested that this group experienced no more re-admissions (OR 1.1, CI 0.7-1.7), no more losses to follow up (OR 1.09, CI 0.6-1.9), and were more successfully discharged on time (OR 0.47, CI 0.3-0.9) compared to long stay or standard care. The data also suggested some evidence that planned short stay patients were no more likely to leave hospital prematurely and had a greater chance of being employed. Data on mental, social and family outcomes could not be summated and there was little or no data on user satisfaction, deaths, violence, criminal behaviour, and costs. REVIEWER'S CONCLUSIONS: The effects of hospital care and the length of stay is important for mental health policy. This review suggests that a planned short stay policy does not encourage a 'revolving door' pattern of admission and disjointed care for people with serious mental illness. More large, well-designed and reported trials are justified. It may be that the 'developing world', where, in some places, the long stay institutions are still functioning, will be able to provide good data that has failed to appear from research in the 'developed world'.
Subject(s)
Length of Stay , Mental Disorders/rehabilitation , Hospitalization , Humans , InstitutionalizationABSTRACT
OBJECTIVE: To determine the effectiveness of planned short hospital stays versus standard care for people with serious mental illness. DESIGN: Systematic review of all randomised controlled trials comparing planned short hospital stay versus long hospital stay or standard care for people with serious mental illness. SUBJECTS: Four trials enrolled 628 patients. MAIN OUTCOMES MEASURES: Relapse; readmission; death (suicides and all causes); violent incidents (self, others, property); lost to follow up; premature discharge; delayed discharge; mental state (not improved); social functioning; patient satisfaction, quality of life, self esteem, and psychological wellbeing; family burden; imprisonment; employment status; independent living; total cost of care; and average length of hospital stay. RESULTS: Patients allocated to planned short hospital stays had no more readmissions (in four trials, odds ratio 0.93, 95% confidence interval 0.66 to 1.29 with no heterogeneity between trials), no more losses to follow up (in three trials of 404 patients, 1.09, 0.62 to 1.91 with no heterogeneity between trials), and more successful discharges on time (in three trials of 404 patients, 0.47, 0.27 to 0.85) than patients allocated long hospital stays or standard care. Some evidence showed that patients allocated planned short hospital stay were no more likely to leave hospital prematurely and had a greater chance of being employed than those allocated long hospital stay or standard care. Data on mental, social, and family outcomes could not be summated, and there were few or no data on patient satisfaction, deaths, violence, criminal behaviour, and costs. CONCLUSION: The effectiveness of care in mental hospitals is important to patients, carers, and policy makers. Despite inadequacies in the data, this review suggests that planned short hospital stays do not encourage a "revolving door" pattern of care for people with serious mental illness and may be more effective than standard care. Further pragmatic trials are needed on the most effective organisation and delivery of care in mental hospitals.
Subject(s)
Hospitals, Psychiatric/statistics & numerical data , Length of Stay/statistics & numerical data , Mental Disorders/therapy , Outcome Assessment, Health Care , Psychiatric Department, Hospital/statistics & numerical data , Activities of Daily Living , Employment , Health Care Costs , Hospitals, Psychiatric/economics , Hospitals, Psychiatric/organization & administration , Humans , Patient Care Planning , Psychiatric Department, Hospital/economics , Psychiatric Department, Hospital/organization & administration , Randomized Controlled Trials as Topic , United KingdomABSTRACT
The effect of tributyltin chloride (TBTC) on rainbow trout (Salmo irideus) hemoglobin I (HbI) and hemoglobin IV (HbIV) was characterized by the steady-state fluorescence of intrinsic and extrinsic fluorescent probes. The fluorescence emission spectrum (lambdaex 280 nm) is greatly increased in intensity by the presence of the organotin in both proteins. Circular dichroism spectra in the same samples show a small decrease in theta222, a measure correlated with the percentage of the alpha-helical content. Morever, important changes in near-UV, Soret, and visible regions of CD were induced by TBTC. The correlation of data obtained with trout hemoglobins (HbI and HbIV) with similar measurements on globins suggests that the presence of heme is necessary for the interaction of the organotin compound with the proteins.
Subject(s)
Hemoglobins, Abnormal/metabolism , Hemoglobins/metabolism , Trialkyltin Compounds/metabolism , Anilino Naphthalenesulfonates/chemistry , Animals , Circular Dichroism , Dose-Response Relationship, Drug , Hydrogen-Ion Concentration , Oncorhynchus mykiss , Spectrometry, Fluorescence , Temperature , Time FactorsABSTRACT
To investigate the molecular mechanisms of the inhibition of Na+,K(+)-adenosine triphosphatase (Na+,K(+)-ATPase) in diabetes mellitus, we incubated Na+,K(+)-ATPase purified from human placenta of six healthy nondiabetic women with plasma from six insulin-dependent diabetic (IDDM) men and six healthy controls and with different concentrations of lysophosphatidylcholine (LPC). We determined the enzyme activity, anthroyl ouabain-binding capacity, dissociation constant (Kd), and average lifetime values (tau) by the static and dynamic fluorescence of anthroyl ouabain. The lipid annulus of the enzyme was studied by static and dynamic fluorescence of 1-(4-trimethylamino-phenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH). Moreover, we studied the lipid microenvironment surrounding the Na+,K(+)-ATPase purified from the placentas of six healthy women and six insulin-dependent diabetic women, determining the percent composition of phospholipids of the lipid annulus. The addition of total and protein-free IDDM plasma to normal Na+,K(+)-ATPase significantly inhibited the enzymatic activity even at the lowest concentration studied (1: 100), whereas the ouabain-binding capacity, Kd, and tau were not affected by IDDM plasma. The fluorescence polarization and lifetime values of TMA-DPH were significantly decreased by diabetic plasma. The incubation of Na+,K(+)-ATPase with LPC caused an inhibition of the enzymatic activity without modifications of the anthroyl ouabain-binding capacity and dissociation constant. The fluorescence polarization and lifetime values of TMA-DPH were significantly decreased by 5 mumol/L LPC. The study of the phospholipids surrounding Na+,K(+)-ATPase demonstrated a significant increase in the percent LPC content in IDDM patients compared with controls together with a concomitant decrease in phosphatidylcholine. These observations indicate that the inhibition caused by diabetic plasma on Na+,K(+)-ATPase is not dependent on a modification of the ouabain-binding site and that it seems to mimic the effect of LPC addition. A link between modification of the lipid moiety of the enzyme and Na+,K(+)-ATPase inhibition might be hypothesized.
Subject(s)
Diabetes Mellitus, Type 1/blood , Lysophosphatidylcholines/pharmacology , Plasma/physiology , Sodium-Potassium-Exchanging ATPase/drug effects , Adult , Case-Control Studies , Female , Fluorescence Polarization , Humans , Male , Phospholipids/analysis , Pregnancy , Sodium-Potassium-Exchanging ATPase/metabolismABSTRACT
BACKGROUND: The action of plasma from women affected by gestational hypertension (GH) on nitric oxide synthase (NOS) activity in cultured human umbilical vein endothelial cells (HUVECs) was evaluated in the present study, together with the effect on cytosolic calcium, on Na+, K+-ATPase activity and on membrane fluidity. METHODS: At 80% confluence, cultured HUVECs were incubated for 3 h at 37 degreesC with fresh culture medium (control samples) or with 20% (v/v) plasma (from five healthy non-pregnant women, five healthy pregnant women and five pregnant women affected by GH). RESULTS: After incubation with GH plasma, we observed a significant reduction in NOS activity, intracellular calcium concentrations and Na+, K+-ATPase activity. CONCLUSIONS: The present work gives further support to the hypothesis that a circulating factor in gestational hypertension, possibly produced by the fetoplacental unit, causes dysfunction of the vascular endothelial cells and NO reduction, resulting in a loss of vascular refractoriness to vasoactive agents.
Subject(s)
Endothelium, Vascular/metabolism , Hypertension/blood , Plasma/physiology , Pregnancy Complications/blood , Adult , Calcium/metabolism , Endothelium, Vascular/drug effects , Female , Humans , Membrane Fluidity/drug effects , Nitric Oxide Synthase/metabolism , Nitric Oxide Synthase Type III , Pregnancy , Sodium-Potassium-Exchanging ATPase/metabolismABSTRACT
The causes of the reduced activity of Na+/K+-adenosine triphosphatase (ATPase) in human diabetes are still the object of controversy. The aim of this work was to investigate the mechanisms of inhibition by means of the study of the Na+/K+-ATPase purified from human placenta. We purified Na+/K+-ATPase from term placentas of six healthy women and six age-matched women with insulin-dependent diabetes mellitus (IDDM) in good metabolic control. The enzymatic activity was reduced in both the microsomal fraction and the purified Na+/K+-ATPase obtained from diabetic women, whereas no difference was found in the number of active molecules determined by anthroyl ouabain binding. The Na+/K+-ATPase purified from women with IDDM did not show any modification in the ouabain affinity or changes in the physicochemical structure of the ouabain binding site investigated by dynamic fluorescence or alterations in lateral diffusion. The activation energy of the enzyme was increased, whereas the tryptophan accessibility of the enzyme was lower in women with IDDM. The fluidity of the lipid anulus of the enzyme was higher in women with IDDM than in control women, as suggested by fluorescence polarization of 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene. The adenosine triphosphate-binding site, investigated by anisotropy decay studies of the fluorescent probe pyrene isothiocyanate, was modified in women with IDDM. It appears that the Na+/K+-ATPase of human placenta is altered in its disposition in IDDM.
Subject(s)
Diabetes Mellitus, Type 1/metabolism , Placenta/enzymology , Pregnancy in Diabetics/enzymology , Sodium-Potassium-Exchanging ATPase/metabolism , Acrylamide , Acrylamides/pharmacology , Adult , Anthracenes/metabolism , Binding Sites , Diffusion , Diphenylhexatriene/analogs & derivatives , Diphenylhexatriene/metabolism , Enzyme Activation , Female , Fluorescence Polarization , Fluorescent Dyes , Humans , Isothiocyanates/metabolism , Kinetics , Membrane Fluidity , Microsomes/enzymology , Ouabain/analogs & derivatives , Ouabain/metabolism , Pregnancy , Sodium-Potassium-Exchanging ATPase/antagonists & inhibitors , Sodium-Potassium-Exchanging ATPase/isolation & purification , Spectrometry, Fluorescence , Tryptophan/metabolismABSTRACT
The stability of trout hemoglobin was examined in the presence of some organotin compounds. Tributyltin chloride (TBTC) and triphenyltin chloride (TPTC) protect HbI most efficently from the oxidation. On the other hand, the same compounds accelerate the precipitation process in HbIV to a great extent. Parahydroxymercuribenzoate (PMB), an agent blocking free SH-groups of the protein, abolished the ability of TPTC to decrease the oxidation rate of HbI.
Subject(s)
Hemoglobins/drug effects , Organotin Compounds/pharmacology , Trialkyltin Compounds/pharmacology , Trout/blood , Animals , Hemoglobins/chemistry , Hemoglobins/metabolism , Organotin Compounds/chemistry , Oxidative Stress , Trialkyltin Compounds/chemistryABSTRACT
In order to investigate the molecular mechanisms of the inhibition of Na+/K+-ATPase in Gestational Hypertension (GH), we incubated Na+/K+-ATPase purified from human placenta of 6 healthy normotensive women with plasma from 6 GH women and 6 healthy controls. We determined the enzyme activity by the method of Esman, and the anthroyl-ouabain-binding capacity, dissociation constant (Kd) and average lifetime values (tau) by the static and dynamic fluorescence of anthroyl-ouabain. The lipid annulus of the enzyme was studied by static and dynamic fluorescence of 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5- hexatriene (TMA-DPH). The addition of total and protein-free GH plasma to normal Na+/K+-ATPase significantly inhibited the enzymatic activity even at the lowest concentration studied (1:100), as well as the ouabain-binding capacity, Kd and tau. GH plasma significantly decreased the fluorescence polarization and lifetime values of TMA-DPH. These observations indicate that the inhibition caused by GH plasma on Na+/K+-ATPase might be due to a reduction of the number of active molecules or a modification of the ouabain-binding site suggesting the existence of digitalis-like factor. A link between the modification of the lipid moiety of the enzyme and the Na+/K+-ATPase inhibition might be hypothesized.
Subject(s)
Enzyme Inhibitors/blood , Hypertension/blood , Placenta/enzymology , Pregnancy Complications, Cardiovascular/blood , Pregnancy/blood , Sodium-Potassium-Exchanging ATPase/metabolism , Adult , Blood Proteins , Female , Humans , Kinetics , Ouabain/metabolism , Reference Values , Sodium-Potassium-Exchanging ATPase/antagonists & inhibitors , Sodium-Potassium-Exchanging ATPase/isolation & purification , Spectrometry, FluorescenceABSTRACT
The HtrA (DegP) protein of Escherichia coli is a heat shock serine protease, essential for cell survival only at temperatures above 42 degrees C. It has been shown by genetic experiments that HtrA is an envelope protease, functioning in the periplasmic space. To clarify the cellular localization of HtrA, E. coli cells were fractionated, and HtrA was not detected by the immunoblotting technique in the periplasm or in the fraction of soluble proteins but was found in the inner membrane. The protein could be partially eluted from the total membrane fraction by a high ionic strength solution, whereas solutions affecting protein conformation released HtrA almost completely. These results, taken together with the evidence showing that HtrA functions in the periplasm, indicate that HtrA is a peripheral membrane protein, localized on the periplasmic side of the inner membrane. As the first step toward solving the problem of HtrA-membrane interactions, the structure of HtrA in the presence of phosphatidylglycerol (PG), phosphatidylethanolamine (PE), or cardiolipin (CL) was analyzed by fluorescence and Fourier-transform infrared spectroscopy. The infrared and fluorescence data indicated an interaction of HtrA with PG and CL but not with PE suspensions. Fluorescence spectroscopy revealed that this interaction was at the level of the polar head group of the phospholipid. In the PG/HtrA system, small changes were observed in the HtrA secondary structure and a remarkable decrease of the thermal stability of the protein, which suggested changes in HtrA tertiary structure. This suggestion was supported by fluorescence data that showed a shift of the fluorescence emission spectrum of HtrA tyrosine residues in the presence of PG and a reduced fluorescence intensity, phenomena not observed in the presence of PE or CL suspensions. Infrared data revealed also that the interaction of HtrA with PG leads to a protection of unfolded protein against aggregation at relatively low temperatures. The conformational changes of HtrA in the presence of PG influenced the proteolytic activity of HtrA by increasing it at the temperatures 37-45 degrees C and inhibiting it at 50-55 degrees C. CL inhibited HtrA activity at all of the temperatures tested.
Subject(s)
Bacterial Proteins/metabolism , Heat-Shock Proteins , Membranes, Artificial , Periplasmic Proteins , Phospholipids/metabolism , Serine Endopeptidases/metabolism , Cardiolipins/metabolism , Cell Membrane/enzymology , Cytoplasm/enzymology , Escherichia coli/enzymology , Phosphatidylethanolamines/metabolism , Phosphatidylglycerols/metabolism , Protein Conformation , Protein Denaturation , Protein Structure, Secondary , Spectrometry, Fluorescence , Spectroscopy, Fourier Transform InfraredABSTRACT
Erythrocytes of Salmo irideus trout were separated in the range from 45 to 65% Percoll, yielding three well-separated different fractions. Steady-state fluorescence of probes embedded in erythrocyte membranes and/or in liposomes from extracted lipids was used to characterize their physicochemical properties. Furthermore, the fluorescence decay of 1,6-diphenyl-1,3,5-hexatriene (DPH), embedded in the same liposomes, was measured by a frequency decay fluorometer. DPH decay was analyzed on the assumption of continuous distribution of lifetimes, for evaluating modifications of membrane microheterogeneity. Significant differences were observed in the parameters measured for the three erythrocyte fractions, possibly connected with the specific lipid composition of the samples.
Subject(s)
Erythrocyte Membrane/chemistry , Animals , Cell Fractionation , Chemical Phenomena , Chemistry, Physical , Cholesterol/blood , Erythrocyte Aging , Fluorescence Polarization , Lipid Peroxides/blood , Liposomes/chemistry , Membrane Fluidity , Membrane Lipids/blood , Phosphatidylcholines/chemistry , Temperature , Trout/bloodABSTRACT
Phase-modulation fluorescence lifetime measurements were used to study the single Trp residue of the Ca(2+)-binding protein S-100a both in the absence and in the presence of Ca2+ and/or Mg2+. Trp fluorescence decay for the protein was satisfactorily described by Lorentzian lifetime distributions centered around two components (approximately 4 ns and 0.5 ns). Lifetime values were unchanged by 2 mM Ca2+, but the fractional intensity associated with longer lifetime increased up to 75%. In the presence of Mg2+, the Ca2+ induced increase of the fractional intensity associated with longer lifetime was only 57%. For the protein in buffer, about the 85% of the recovered anisotropy was associated to a rotational correlation time of 6.7 ns. After the addition of Ca2+, this value was increased to 16.08 ns. In the presence of Mg2+, Ca+2 increased the rotational correlation time to 33.75 ns. Similar studies were performed with S-100a interacting with egg phosphatidylcholine vesicles (SUV). Our data suggest that the conformation of the protein may be influenced by structural features of the lipidic membrane. Moreover, data obtained in the presence of Mg2+ indicate some interaction between lipids and S-100, likely mediated by this ion.
Subject(s)
Calcium-Binding Proteins/analysis , Calcium/pharmacology , Magnesium/pharmacology , Phosphatidylcholines/pharmacology , S100 Proteins , Animals , Brain Chemistry , Calcium-Binding Proteins/chemistry , Cattle , Fluorescence Polarization , Intracellular Membranes/chemistry , Lipids/analysis , Ovum/chemistry , S100A12 Protein , Time FactorsABSTRACT
1. Na+,K(+)-ATPase is the membrane enzyme catalysing the active transport of Na+ and K+ across the plasma membrane of animal cells. A reduced activity of Na+,K(+)-ATPase has been described in gestational hypertension in a variety of cell types, in agreement with the hypothesis that gestational hypertension can induce membrane transport modifications similar to those reported for essential hypertension. The causes of the reduced Na+,K(+)-ATPase activity are still debated. 2. The aim of the present work was to investigate the molecular mechanism of the reduced enzymic activity in gestational hypertension using as a model Na+,K(+)-ATPase purified from human placenta. Na+,K(+)-ATPase obtained from term placentas of eight healthy pregnant women and eight age-matched women with gestational hypertension was purified as previously described. 3. We observed in gestational hypertension: (i) a significant increase in the activation energies above transition temperature; (ii) a significant decrease in the fluorescence polarization of 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene (i.e. increased fluidity) and an increase in the mean lifetime (modified hydrophobicity); (iii) a lower Kq, suggesting an enzymic structural modification; and (iv) an increased mean lifetime and rotational relaxation time of pyrene isothiocyanate, indicating a modified ATP binding site.
Subject(s)
Hypertension/enzymology , Placenta/enzymology , Pregnancy Complications, Cardiovascular/enzymology , Sodium-Potassium-Exchanging ATPase/metabolism , Adult , Biophysical Phenomena , Biophysics , Female , Fluorescence Polarization , Humans , Pregnancy , Temperature , TryptophanABSTRACT
The case of a women with polycystic ovary disease and delusional disorder is reported. During the course of her paranoid illness she developed firstly "homosexual' erotomania and later "heterosexual' erotomania. We suggest that this is a rare presentation of delusional disorder.
Subject(s)
Bipolar Disorder/psychology , Bisexuality/psychology , Delusions/psychology , Erotica , Neurocognitive Disorders/psychology , Polycystic Ovary Syndrome/psychology , Adult , Bipolar Disorder/diagnosis , Chronic Disease , Delusions/diagnosis , Female , Humans , Neurocognitive Disorders/diagnosis , Polycystic Ovary Syndrome/diagnosis , SyndromeABSTRACT
The effect of N-lauroylethanolamine (N-LEA) and N-oleoylethanolamine (N-OEA) on the thermal behaviour of fully hydrated egg phosphatidylethanolamine (TPE) was investigated by the steady-state fluorescence of 2-dimethylamino-(6-lauroyl)-naphtalene (laurdan) and 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH). The parameter generalised polarisation (GP), calculated by exciting laurdan at 340 and 410 nm, revealed the gel to liquid crystalline lamellar (L alpha) as well as the L alpha to inverse hexagonal (HII) phase transitions of TPE. The L alpha to HII phase transition was not detected in TPE/N-OEA system, probably because of the formation of an intermediate Q224 cubic phase. The formation of Q224 phase in TPE/N-OEA and TPE/N-LEA systems was previously demonstrated by X-ray diffraction, but neither laurdan generalised polarisation nor TMA-DPH steady-state fluorescence anisotropy measurements revealed the presence of this phase. It is suggested that the lack of detection of the cubic phase is probably due to the similarity in dynamic characteristics and hydration levels of phospholipid headgroups in the bilayer and cubic phases.