Rapid and fully automated bacterial pathogen detection on a centrifugal-microfluidic LabDisk using highly sensitive nested PCR with integrated sample preparation.

Diagnosis of infectious diseases suffers from long turnaround times for gold standard culture-based identification of bacterial pathogens, therefore impeding timely specific antimicrobial treatment based on laboratory evidence. Rapid molecular diagnostics-based technologies enable detection of microorganisms within hours however cumbersome workflows and complex equipment still prevent their widespread use in the routine clinical microbiology setting. We developed a centrifugal-microfluidic "LabDisk" system for rapid and highly-sensitive pathogen detection on a point-of-care analyser. The unit-use LabDisk with pre-stored reagents features fully automated and integrated DNA extraction, consensus multiplex PCR pre-amplification and geometrically-multiplexed species-specific real-time PCR. Processing merely requires loading of the sample and DNA extraction reagents with minimal hands-on time of approximately 5 min. We demonstrate detection of as few as 3 colony-forming-units (cfu) of Staphylococcus warneri, 200 cfu of Streptococcus agalactiae, 5 cfu of Escherichia coli and 2 cfu of Haemophilus influenzae in a 200 µL serum sample. The turnaround time of the complete analysis from "sample-to-result" was 3 h and 45 min. The LabDisk consequently provides an easy-to-use molecular diagnostic platform for rapid and highly-sensitive detection of bacterial pathogens without requiring major hands-on time and complex laboratory instrumentation.

Sphingosine 1-phosphate mobilizes sequestered calcium, activates calcium entry, and stimulates deoxyribonucleic acid synthesis in thyroid FRTL-5 cells.

Sphingosine 1-phosphate (SPP) potently mobilizes sequestered calcium and is a mitogen in several cell types. In the present investigation, we have evaluated the effect of SPP on intracellular free calcium concentration ([Ca2+]i) and synthesis of DNA in thyroid FRTL-5 cells. SPP rapidly and transiently mobilized sequestered calcium and stimulated entry of extracellular calcium. The entry of calcium, but not the mobilization, was in part inhibited by pretreatment with pertussis toxin (Ptx), and by activation of protein kinase C. SPP did not stimulate the production of inositol 1,4,5-trisphosphate. SPP stimulated the incorporation of 3H-thymidine in a time- and dose-dependent manner. The effect was not inhibited by Ptx. Furthermore, SPP stimulated the activation of the proto-oncogene c-fos. SPP rapidly tyrosine-phosphorylated an approximately 66 kDa protein. This phosphorylation persisted for at least 1 h. Pretreatment of the cells with genistein abolished the SPP-evoked tyrosine phosphorylation, and attenuated the SPP-evoked increase in [Ca2+]i. Furthermore, the SPP-evoked activation of Na+-H+ exchange was inhibited by genistein. The phosphorylation was not attenuated by pretreatment of the cells with Ptx. SPP per se did not affect cellular cAMP levels but attenuated the TSH-evoked increase in cAMP. As the effect of SPP might be due to activation of phospholipase D, we tested whether phosphatidic acid (PA) mobilized calcium or stimulated the incorporation of 3H-thymidine. PA mobilized sequestered calcium but did not stimulate calcium entry. PA very modestly enhanced the incorporation of 3H-thymidine. Our results suggest, that SPP stimulates DNA synthesis and activates entry of calcium in FRTL-5 cells. The effect on calcium entry appears to be dependent, at least in part, on one or several tyrosine kinases.

Evidence for the role of hepatic endothelial lipase in the metabolism of plasma high density lipoprotein2 in man.

Previous animal studies have shown that the heparin-releasable hepatic lipase (HL) is located on the luminal surface of the liver endothelial cells and may have a function in the removal of high density lipoprotein lipids from plasma. We therefore examined the relationship between plasma HDL levels and the HL activity of postheparin plasma in a group of young, very fit men who were living under strictly controlled comparable conditions (military academy studients). HDL2 cholesterol, HDL2 phospholipid and HDL2 protein concentrations each showed a highly significant negative correlation with postheparin HL activity. A similar but slightly lower inverse relationship was also present between total HDL lipids and HL activity, whereas no correlation could be observed between any of the HDL3 lipids and HL activity. The cholesterol/protein ratio of HDL2 correlated negatively with the HL activity. These results support the hypothesis that the hepatic endothelial lipase has a physiological role in the degradation and removal of circulating HDL2.

Plasma high density lipoproteins HDL2, HDL3 and postheparin plasma lipases in relation to parameters of physical fitness.

A number of studies has shown that the plasma levels of high density lipoprotein (HDL) are increased by regular aerobic exercise. The plasma HDL, particularly HDL2, is regulated by the activity of 2 endothelial lipases, viz. lipoprotein lipase (LPL) and hepatic lipase (HL), which both can be assayed in postheparin plasma. In the present study the plasma levels of HDL2 and HDL3 cholesterol and the postheparin plasma lipase activities were related to parameters of physical fitness obtained from a pulse conducted maximal bicycle ergometer test. There was a significant positive correlation between HDL2 cholesterol and physical fitness (r = 0.52, P less than 0.01). On the other hand, the postheparin plasma hepatic lipase activity showed a significant negative correlation to physical fitness (r = -0.57, P less than 0.01). The HDL2 cholesterol was inversely correlated with the HL activity (r = 0.57, P less than 0.001). Application of partial correlation analysis to the data showed that the relationship between HDL2 cholesterol and fitness disappeared by keeping the HL activity constant whereas the correlation between HDL2 and HL was not influenced by fitness. The relation of HDL2 to fitness was independent in body fat and basal plasma insulin level; in addition the relationship between HL and fitness was not accounted for by body fatness. No relationship was found between physical fitness and LPL activity or between HDL3 and fitness. The results support the hypothesis that hepatic endothelial lipase has a role in the regulation of plasma HDL2 cholesterol and that the activity of this enzyme decreases upon increase of physical fitness.

Suicide risk in schizophrenia: an analysis of 17 consecutive suicides.

The aim of this study was to investigate interactional factors related to the recognition of suicide risk in patients with schizophrenia. The study focused on 17 schizophrenia patients who had committed suicide during the National Suicide Prevention Project in Finland between April 1, 1987, and March 31, 1988, in the province of Kuopio. Consensus case reports were assembled by using the psychological autopsy method. Study methods included structured and in-depth interviews of next of kin and interviews of health care or social services workers who had treated the suicide victims. Male and female patients with schizophrenia committed suicide in equal proportions. Most had suffered from schizophrenia for more than 15 years; all but one had been receiving psychiatric treatment at the time of suicide. Retrospective assessment indicated that 59 percent of the patients were clinically depressed at the time of suicide. In 76 percent of the cases, the mental health professionals involved in treatment had not believed that there was a risk of suicide during their last contact with the patient. In 29 percent of the cases, the patient's paranoid ideas concerning treatment personnel had increased. Patients' withdrawal from human relationships because of depression was related to loss of the treatment professionals' concern for the patients. The findings in this descriptive study suggest that withdrawal by a patient with schizophrenia and an increase in the patient's paranoid behavior should be regarded as signals of risk of suicide.

Is it possible to adapt to the suicide of a close individual? Results of a 10-year prospective follow-up study.

We conducted a prospective ten-year follow-up study on the consequences of a suicide among relatives or other individuals close to the suicide victim. Baseline interviews (n = 104) were carried out during the National Suicide Prevention Project in Finland from 1987 to 1988. Semi-structured follow-up interviews including the 12-item General Health Questionnaire and the 21-item Beck Depression Inventory were carried out ten years later (n = 64, 64% of those still alive). Mental symptoms were reported to have been common after suicide but they had subsided during three years. Mental symptoms had been common especially among children after their parent's suicide. At baseline mothers had had depressive symptoms more often than others, but at follow-up there was no statistical difference. Life changes associated with the suicide were common, and they could be unfavourable as well as favourable. At the time of the follow-up study, half of the interviewees felt that baseline interviews had helped them to adjust to the suicide.

Line-shape distortions in misaligned cube corner interferometers.

Line-shape distortions caused by the misalignment of the moving cube mirror in Fourier transform spectrometers have been described. A method of studying and correcting these distortions is presented. By using this method we can estimate the accuracy of the line position, which is especially important in high-resolution Fourier transform spectroscopy. The method is verified in simulations, and in practice it has been used to align the Oulu Fourier transform spectrometer.

Spectral line-shape distortions in Michelson interferometers due to off-focus radiation source.

When the radiation source in a Michelson interferometer is placed in front of or behind the focal plane of the collimator, distortions arise in the spectral line shapes. The appearance and behavior of these distortions in a cube-corner interferometer are treated in this work. We shall also present a simple and fast method to calculate the true line position of a distorted line, and an easy rule of thumb to connect the off-focus shift and the amount of distortion. Calculation of the phase-error curves in the signal domain is treated as well. This calculation gives a possibility of correcting the distorted interferogram.

Adenosine inhibits DNA synthesis stimulated with TSH, insulin, and phorbol 12-myristate 13-acetate in rat thyroid FRTL-5 cells.

Adenosine has been shown to modulate cell proliferation in FRTL-5 thyroid cells, although the mechanisms by which this interaction occurs is still unclear. In the present study we investigated the effects of adenosine on the 3H-thymidine incorporation, cell cycle kinetics, and expression of the transcription factor c-Fos in cells stimulated via three different mitogenic pathways, i.e., by thyroid stimulating hormone (TSH) [adenosine 3',5'-cyclic monophosphate(cAMP)], insulin (tyrosine kinase), or phorbol 12-myristate 13-acetate (protein kinase C). Addition of adenosine to cells grown in medium containing hormones and serum did not inhibit the incorporation of 3H-thymidine. If adenosine was added to hormone-deprived cells together with any of the tested mitogens, the stimulation of the 3H-thymidine incorporation was inhibited in a dose-dependent manner. The inhibition was significantly lower when the cells were preincubated with TSH or insulin for 48 h. Flow cytometric studies showed that adenosine evoked an inhibition of the cells in the G0/G1 phase. Submaximal doses of adenosine (10 nM-10 microM) were able to induce c-Fos expression in FRTL-5 cells. However, the mitogen-induced expression of c-Fos was not reduced by maximal dose of adenosine (100 microM). The effect of adenosine on DNA synthesis was not dependent on pertussis toxin-sensitive G-proteins. In addition, adenosine A1- or A2- receptor antagonists did not block the effect of adenosine. The effect of adenosine was abolished by treatment of the cells with adenosine deaminase, suggesting that the observed effect was not mediated by a metabolite of adenosine. The results suggest that adenosine is an effective blocker of mitogen-evoked DNA synthesis of FRTL-5 cells, provided that adenosine is administered simultaneously with the mitogen.

Labetalol and pindolol in the treatment of hypertension: a comparative study.

Labetalol, a new hypotensive drug with both alpha- and beta-adrenoceptor antagonist properties, was compared with pindolol in a double blind cross-over study lasting 15 weeks. Both drugs caused a significant drop on blood pressure in both the standing and the supine position. The effect was seen after only two weeks treatment. The effect on the pulse rate was moderate. Three patients on pindolol (but none on labetalol) had to withdraw from the trial for medical reasons. In general the side effects recorded for labetalol and pindolol were mild. No postural hypotension was noticed with the moderate doses used in the study. Labetalol seems to be at least as effective as pindolol in mild to moderate hypertension.

Suicide mortality in Finland during an economic cycle, 1985-1995.

This is a study on associations between suicide mortality, unemployment, divorce rate and mean alcohol consumption during an economic cycle in Finland, from 1985 to 1995. Data on annual suicide mortality, gross domestic product, unemployment, divorce rate and mean alcohol consumption were collected from official Finnish statistics. Regression analyses using a correction for serial autocorrelation were performed. Suicide mortality in both males and females increased during an economic upswing from 1985 to 1990 and decreased during an economic recession from 1990 to 1995. Suicide mortality was not associated with unemployment or divorce rate. These results are opposed to those of many previous studies. However, a significant association was found between male suicide mortality and mean alcohol consumption, in accord with other studies. In conclusion, the results suggest diversity in associations between suicide mortality, socioeconomic factors and alcohol consumption.

Is alexithymia a permanent feature in depressed patients? Results from a 6-month follow-up study.

BACKGROUND: A six-month follow-up study was conducted to determine whether alexithymia is a permanent feature in 169 depressed outpatients. METHODS: Diagnosis of depression was confirmed by means of the Structured Clinical Interview for DSM-III-R (SCID-I). Alexithymia was screened using the 20-item version of the Toronto Alexithymia Scale (TAS-20) and severity of depression was assessed using the 21-item Beck Depression Inventory (BDI). RESULTS: Almost 40% of the patients were considered alexithymic at baseline, but only 23% at follow-up. Alexithymic patients were more often moderately or severely depressed than other patients in both study phases. The BDI scores explained 23% (at baseline) and 42% (at follow-up) of the variation in TAS-20 scores. The decrease in the TAS-20 scores was associated with a concurrent decrease in BDI scores. CONCLUSIONS: Alexithymic patients with depressive disorders do not appear to form a stable group. On the contrary, alexithymia seems to change as a function of depression. In the light of these results, alexithymia appears not to be a stable personality trait among depressed patients, and furthermore, it seems possible that alexithymic features respond to psychiatric treatment.

Suicidal ideation in the Finnish general population. A 12-month follow-up study.

BACKGROUND: The epidemiology of suicidal ideation has remained a relatively unstudied area. The aim of this study was to investigate the incidence, prevalence and persistence of and recovery from suicidal ideation in a sample of the Finnish general population. METHODS: Postal questionnaires including the Beck Depression Inventory (BDI) were mailed to the study subjects (n = 1,593) at baseline and on follow-up after 12 months. The suicidality item of BDI was used to screen suicidal ideation. RESULTS: The 12-month incidence (4.6%) and prevalence of suicidal ideation (14.7%) were higher in men than in women (3.1% and 9.2%, respectively). Sixty-nine per cent of those men and 59% of those women who had suicidal ideation at baseline continued to have suicidal thoughts on follow-up. Suicidal ideation and the severity of depression associated strongly. Over half of the men and women who had persistent suicidal ideation had not visited any health services for help with psychological distress during the 12-month follow-up period. Daily smoking associated with a decreased probability of recovery from suicidal ideation. CONCLUSIONS: Suicidal ideation is common and persistent at the population level. Seeking professional help for depression with suicidal tendencies is disproportionately low in relation to the high prevalence of suicidal ideation.

Suppression of the TSH response to TRH by thyroxine therapy in differentiated thyroid carcinoma patients.

Absent response of serum thyrotrophin (TSH) after stimulation with 200 micrograms synthetic thyrotrophin-releasing hormone (TRH) was used as a criterion of adequate suppression of TSH in the treatment of thyroid carcinoma patients with thyroxine. The mean causing total suppression of the response was 223 micrograms of thyroxine per day. At this dose level about 40% of the patients had serum thyroxine concentrations above the upper reference interval and only 10% had elevated triiodothyronine concentrations. In some patients the TSH response to TRH varied between absent and low normal when tested at long intervals. The ideal dose of thyroxine is obviously slightly higher than the smallest one causing total suppression of the TSH response to TRH, i.e. about 250 micrograms a day. The individual dose must be found using the TRH stimulation test because serum thyroid hormone levels cannot be used as a guideline for adequate dosage. In some patients the thyroid remnant of apparently normal thyroid tissue was not totally suppressed although the thyroxine dose was definitely above the level causing suppression of the response to TRH.

Debt and suicidal behaviour in the Finnish general population.

We studied the significance of debt as a risk factor for suicidal ideation and suicide attempts in a nation-wide sample (n = 4868) of the Finnish general population. Those experiencing difficulties in repaying their debts more often than others (37 vs. 16%, P<0.001) had a probable minor mental disorder (GHQ-12 score > or =3). Nevertheless, difficulties in repaying debts were found to be a factor independently associated with suicidal ideation (OR, 2.8; 95% CI, 1.9-4.2). The clinical implication of these findings is that individuals experiencing difficulties in repaying their debts may require psychiatric evaluation as well as socio-economic counselling.

Risk factors for suicidal ideation in psychiatric patients.

Sociodemographic and clinical risk factors for suicidal ideation have been less studied than risk factors for parasuicide and suicide. No reports on associations between therapy satisfaction and suicidal ideation among psychiatric patients have been published. In this study we compared a group of patients with suicidal ideation (n = 84) with a randomly selected group of nonsuicidal patients (n = 166) in community-based psychiatric services. Patients with suicidal ideation felt a need for psychiatric treatment more often than nonsuicidal patients. They were also more likely to receive antidepressive medication, and weekly therapy sessions were more common among them. A wish to change therapist (OR 15.6, 95% CI 3.6-67.8), hopeless future orientation (OR 14.8, 95% CI 4.5-48.9), severe depression as evaluated by the Beck Depression Inventory (OR 14.0, 95% CI 4.3-45.2) and dysthymia (OR 12.8, 95% CI 1.7-97.3) were the factors most strongly associated with suicidal ideation in multivariate analysis. A wish to change therapist is an expression of therapy dissatisfaction, which may therefore be among the factors most strongly associated with suicidal ideation in psychiatric patients. To help prevent suicidality among psychiatric patients special attention to therapy factors is needed.

The effect of psychotropic drugs on the TSH-response to thyroliberin (TRH).

The effect of chlorpromazine (CPZ), thioridazine (TR), L-dopa, fusaric acid (FA), pimozide (PZ) and propranolol (P) in the i.v. thyrotropin-releasing hormone (TRH) stimulation test was studied in 75 subjects. It was found that chronic treatment with CPZ, TR and L-dopa inhibits the response to TRH. FA may to some extent counteract the effect of L-dopa. The results indicate that the response of the pituitary to TRH as regards the secretion of TSH is mainly regulated by norepinephrinergic alpha-receptors.