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At least 433 records · Page 24Linked to original sources
PubMed1989

Abnormalities of thyrotrophin (TSH) evening rise and pulsatile release in haemodialysis patients: evidence for hypothalamic-pituitary changes in chronic renal failure.

Using a sensitive enzyme amplified immunoassay for TSH, the evening rise and pulsatile release of TSH were studied in 10 men with chronic renal failure treated by haemodialysis. Compared to euthyroid male controls the evening rise of TSH was attenuated (median 0.066 vs 0.195 mU/l/h, P less than 0.01) and the rate of rise correlated with the TSH response to TRH (r = 0.93, P less than 0.001). All subjects showed TSH pulsatility in at least one method of data analysis but the less sensitive incremental method showed no significant difference in pulse frequency and amplitude between the two groups. However, with time series analysis, periodicity was shorter (median 45 vs 95 min, P = 0.013) and pulse amplitude smaller (median 0.06 vs 0.175 mU/l, P = 0.017) in renal patients. Pulse amplitude, but not periodicity, correlated with the TSH response to TRH (r = 0.68, P less than 0.05). In addition, serum total thyroxine, free thyroxine and free triiodothyronine concentrations were reduced, while serum prolactin and 17 beta-oestradiol concentrations were raised. These changes in TSH evening surge and pulsatile release may contribute to the reduction in thyroidal hormone concentrations seen in renal failure and emphasize the value of sensitive methods of hormone and pulse data analysis.

Adult↗
PubMed2004

[Hemolysis in blood samples. Assessment in 3 extraction systems].

Blood extraction by means of a vacuum system is a technique which has been employed in our field for some time, although this method does not receive all the popularity it should. This method offers something which the classical syringe can not: greater security. Proper blood extraction leads to a decrease in problems/errors which are susceptible to appear during the pre-analysis period. The use of a system which combine the characteristics of a vacuum with those of classical syringes, such as the Holdex de Vacuette (Bio Greiner, Madrid, España-H) can help to change this negative tendency. However, the special characteristics of this system (bendable to adapt to the site of a phlebotomy with the tube used to extract a blood sample) could cause an increase in the hemolysis in samples due to theoretically greater difficulty blood flow has in this circuit in comparison with other usual procedures in our filed (syringe and vacuum system. To test this assumption, 512 patients who had been asked to undergo a routine biochemical analysis were randomly chosen. These patients were divided into 3 groups, one for each extraction method used; 171 used Holdex de Vacuette(H), 168 used Vacutainer (V), and 173 used a syringe(J). The presence of hemolysis in these tubes was tested and the number of cases was compared to those detected among all the samples carried out during the study period. No statistically significant differences were discovered either among the 3 extraction systems used (H 2.1 or 16%, V 2.1 or 15%, J 4.2 or 23%) nor with the all samples analyzed (39, 2.02% total; 8, 1.56% study group, p < 0.001). These results let us conclude that the number of cases of hemolysis which occur when using Holdex is not greater than the number which occur when using other extraction systems and this method can be a good alternative to the conventional vacuum system.

Adolescent↗
PubMed2007

Analysis of polyhydroxyalkanoate (PHA) synthase gene in activated sludge that produces PHA containing 3-hydroxy-2-methylvalerate.

The identification of phaC which encodes PHA synthase, that is involved in the formation of polyhydroxyalkanoate (PHA) containing 3-hydroxy-2-methylvalerate (3H2MV), was attempted. As of now, PHA containing 3H2MV has been reported to be produced only by mixed microbial cultures in activated sludge, but no pure bacterial cultures. A laboratory-scale activated sludge process was operated for 67 days. During the operation of the activated sludge process, its capacity to produce PHA containing 3H2MV, and the diversity of the partial phaC genes in the activated sludge microorganisms were monitored periodically. Analysis of the partial phaC genes was conducted by PCR followed by cloning and DNA sequencing, or by PCR followed by terminal-restriction fragment length polymorphism (T-RFLP). The cloning-sequencing of the 263 clones gave 11 distinct genetic groups (GGs). All of the genetic groups had similarities to known phaC higher than 48%, and one of them had similarity as high as 96% to that of Alcaligenes sp. The behavior of each of the genetic groups during the operation of the activated sludge process was monitored by the T-RFLP method. The restriction enzyme AccII, with the help of MboI, enabled the monitoring of each of the genetic groups. One of the genetic groups was found to have a strong correlation with the capability of the activated sludge to produce PHA containing 3H2MV, and its DNA sequence together with its amino acid sequence are reported.

Acyltransferases↗
PubMed2001

Effect of technical settings and semen handling upon motility characteristics of dog spermatozoa measured using computer-aided sperm analysis.

Technical aspects of computer-aided sperm analysis and the influence of semen preparation were investigated for their effect on the measured motility characteristics of dog spermatozoa. Altering the internal image settings significantly influenced the measured motility by changing the ability of the computer to recognize spermatozoa. Similarly, the use of a longer minimum track point (the minimum length of sperm track detected before analysis) resulted in failure to detect some of the faster moving spermatozoa. There was a clear interaction between the search radius (the threshold distance below which objects are linked together) and the minimum track point. A 1 min analysis period was required to eliminate reduced motility as a result of sample deterioration upon the microscope stage. The dilution of semen to between 1:10 and 1:20 was necessary to allow accurate detection of sperm motility; however, such dilution significantly altered the motility characteristics of spermatozoa. The influence of viscosity and ionic composition of the media was confirmed by comparing dilution in seminal plasma with dilution in iso-viscous methylcellulose and iso-osmotic saline, respectively. Analysis temperature had a significant influence on sperm motility, although values were most constant within the range of 25-45 degrees C. Extremes of temperature had marked deleterious effects. Careful selection of internal image settings, the minimal track point and search radius, and the analysis time are essential for accurate detection of sperm motility. Moreover, dilution of spermatozoa per se, and dilution with media of different viscosities and ionic compositions can alter the sperm motility. Once these aspects of computer image analysis are determined for each system, the method can achieve a high degree of repeatability with interanalysis coefficients of variation of < 12%, and intra-analysis coefficients of variation of < 3% for most parameters.

Animals↗
PubMed2005

Influence of the very polluted inputs of the Tinto-Odiel system on the adjacent littoral sediments of southwestern Spain: a statistical approach.

A spatial and temporal analysis (period 1990-2003) of 15 sampling points distributed along the southwestern Spanish coast permits to delimitate the influence area of the extremely polluted discharges coming from the Tinto-Odiel system in the bottom sediments of the adjacent littoral area. As, Cu, Pb and Zn are the main heavy metals transported by the freshwater runoffs toward the shallow shelf and present very high negative (r < -0.7) and significant (p < 0.001) correlations with the distance to the estuarine mouth. The statistical analysis (index of geoaccumulation, Pearson correlation matrix, cluster analysis) of their concentrations in the littoral sediments located between the Guadiana and Guadalquivir mouths delimitates three zones: (a) Zone 1 (from the estuarine mouth to 6 km to the east), characterized by moderate to strongly polluted bottom sediments and main responsible of the mean annual variations of the former heavy metals in the area studied; (b) Zone 2 (from 21.2 km to the west to 29 km to the east), characterized by moderate pollution levels; and (c) Zone 3, located near the Guadiana and Guadalquivir mouths, with very low As-Cu-Pb contents and unpolluted to moderately levels of Zn due to urban sewages or the presence of local low mobility areas for this element.

Data Interpretation, Statistical↗
PubMed1984

Rotavirus carriage, asymptomatic infection, and disease in the first two years of life. II. Serological response.

Serological response to rotavirus and virus shedding were prospectively studied in 179 children (neonatal to 24-month-old) upon admission to a hospital during an 11-month period. Analysis of the evolution of IgG and IgM ELISA titers revealed 24 cases of rotaviral disease (serological response and diarrhea), 13 cases of asymptomatic infection (serological response and no diarrhea), 36 cases of virus carriage (absence of a serological response), three cases of past infection, and six possible cases of nosocomial infection. Rotaviral disease was encountered two out of three times and was characterized by diarrhea associated with fever and vomiting. Asymptomatic rotaviral infection and disease, observed from the neonatal period onwards, affected 2% of neonates, 20% of one- to six-month-old children, and 37% of 7-24-month-old children. In contrast, virus carriage occurred in 27%, 19%, and 14% of those children respectively. Altogether these results indicate that during the period 1-24 months of age, when asymptomatic rotaviral infection and disease were prevalent, approximately two of 10 children had rotaviral disease, one of 10 had asymptomatic infection, two of 10 were virus carriers, and five of 10 were not infected with rotavirus.

Aging↗
PubMed1997

Donor type microchimerism is an infrequent event following liver transplantation and is not associated with graft acceptance.

Donor-type microchimerism, the presence of a minority population of donor-derived haematopoietic cells following solid organ transplantation, has been postulated as a mechanism for induction of donor-specific graft tolerance. The stability, frequency, and relevance of microchimerism with respect to long-term outcome, however, remains uncertain. Using a polymerase chain reaction (PCR)-based method of microsatellite analysis of highly polymorphic short tandem repeat sequences (STRs) to detect donor-type cells, DNA from 11 patients was analyzed prospectively at specific time points for 12 months following liver transplantation, and from a further six patients retrospectively 2 years after liver transplantation. Using a panel of STRs, transient peripheral blood donor microchimerism was detected in 2 of 11 patients at a single time-point following transplantation, but persistent evidence of donor-derived cells was not observed during the study period. Analysis of DNA extracted from skin and duodenum in two patients likewise failed to show donor-type cells at these sites. None of the six patients in the retrospective arm showed donor microchimerism, resulting in an overall detection rate of 1.58%. These results suggest that donor microchimerism following liver transplantation is an infrequent event, and that the generation of graft tolerance is independent of microchimerism.

Chimera↗
PubMed1998

Coupling of sympathetic and somatic motor outflows from the spinal cord in a perfused preparation of adult mouse in vitro.

1. The relationship between sympathetic and somatic motor outflows from thoraco-lumbar spinal cord was investigated in a novel arterially perfused trunk-hindquarters preparation of adult mouse. 2. Ongoing activity was present in both somatic motor (obturator, sciatic or femoral nerves) and sympathetic outflows (either renal nerve or abdominal sympathetic chain). Sympathetic activity was rhythmic with bursts frequencies of 0.6-2.2 Hz. No obvious rhythmic activity was found in the somatic motor outflow. There were periods during which sympathetic and somatic motor activity were correlated. 3. Addition of NMDA (20-80 microM) to the perfusate elicited repetitive burst discharges in the somatic motor outflow which were sometimes rhythmic. The frequency of these burst discharges/rhythmic activity varied between preparations but in all cases increased with increasing NMDA concentration. 4. NMDA induced burst discharges in the sympathetic outflow. This bursting activity was of the same frequency as the somatic motor outflow and the two were coupled as revealed by correlation analysis. Periods of coupling persisted for up to 3 min. 5. Administration of hexamethonium (300 microM), to block sympathetic ganglionic transmission, had no effect on the somatic motor activity but severely attenuated sympathetic nerve discharge. 6. The thoraco-sacral cord therefore has the neuronal machinery necessary for generating and coupling activity in somatic motor and sympathetic outflows. Our findings indicate a dynamic control over the degree of coupling. We discuss that the synchronization of these neural outflows reflects either coupling between two independent mechanisms or the presence of a common synaptic driver impinging on both somatic motor and sympathetic neurones.

Animals↗
PubMed1991

Naloxone increases the frequency of the electrical activity of luteinizing hormone-releasing hormone pulse generator in long-term ovariectomized rats.

Characteristic increases in neuronal activity accompanied by the initiation of each luteinizing hormone (LH) pulse have been successfully recorded in freely moving ovariectomized rats by means of multiunit activity (MUA) recording techniques. In the present study, the effect of the opioid receptor antagonist naloxone on this neuronal activity was examined in rats ovariectomized for 6-10 weeks. Electrodes were chronically implanted into the medial basal hypothalamus (n = 78) or the medial preoptic area (n = 29). During the MUA recording, blood samples were taken through an indwelling atrial cannula at 3- or 6-min intervals to determine the serum LH concentration. Naloxone was administered intravenously for 1 h by either intermittent injection every 6 min (0.05 mg/kg, 10 times) or continuous infusion (0.1, 0.2 and 0.5 mg/kg/h). Explosive rises in the MUA (volleys) associated with the initiation of LH pulse were recorded in 7 animals in which the tips of electrodes were located in the arcuate nucleus-median eminence region. In 5 animals studied, the mean (+/- SE) duration and interval of the MUA volleys were 2.1 +/- 0.1 and 22.1 +/- 0.9 min, respectively. Naloxone given every 6 min significantly increased the duration to 2.6 +/- 0.1 min, and decreased the interval to 9.5 +/- 0.9 min. This naloxone treatment disturbed the clear pulsatility of LH secretion observed in the pretreatment control period. Analysis of the effects of naloxone infused at 3 different doses revealed that the facilitatory effects of naloxone on the duration and frequency of the MUA volleys were dose-dependent.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2003

Induction of myocardial biglycan in heart failure in rats--an extracellular matrix component targeted by AT(1) receptor antagonism.

OBJECTIVE: Cardiac remodelling associated with congestive heart failure typically involves dilatation of the ventricular cavities, cardiomyocyte hypertrophy and alterations of extracellular matrix. Biglycan is an extracellular proteoglycan with several recently appreciated functions including cell adhesion, collagen fibril assembly, and growth factor interactions. The aims of this study were to investigate the regulation of biglycan expression and to elucidate the site(s) of synthesis of biglycan in myocardial tissue in an experimental model of heart failure (HF). METHODS: Myocardial tissue samples were obtained from rats with myocardial infarction (MI) subsequent to ligation of the left coronary artery. Northern blot analysis and real-time quantitative RT-PCR were employed to investigate mRNA levels. The cellular distribution of biglycan was analysed by in situ hybridisation and immunohistochemistry. RESULTS: Myocardial biglycan mRNA levels in non-ischemic tissue of both left and right ventricles of heart failure rats were substantially elevated as compared to sham-operated rats. Although expression levels peaked 7 days after MI (13-fold increase compared to the sham group, P<0.05), substantial elevations of biglycan mRNA were observed throughout the study period. Analysis of cellular distribution revealed that biglycan expression was confined to myocardial fibroblasts and vascular endothelial cells. In cardiac fibroblasts isolated from failing hearts, biglycan mRNA levels were markedly elevated compared with fibroblasts from sham-operated rats. In addition, in rats with ischemic heart failure treatment with the AT(1) receptor antagonist losartan (12.5 mg.kg(-1) b.i.d. per os, for 25 days) prevented the increase of myocardial biglycan as well as TGF-beta(1) mRNA. CONCLUSION: This report demonstrates global induction of myocardial biglycan mRNA in heart failure. Myocardial biglycan expression could be targeted by AT(1) receptor antagonism, an intervention well documented to halt cardiac remodelling in heart failure. Furthermore, the study provides evidence that angiotensin II is a regulator of biglycan expression in cardiac fibroblasts.

Angiotensin II Type 1 Receptor Blockers↗
PubMed1985

Macromolecules released into the culture medium during the vegetative cell cycle of the unicellular green alga Chlamydomonas reinhardii.

The culture medium of growing Chlamydomonas reinhardii cells contains hydroxyproline-rich glycoproteins, which are mainly liberated during release of the zoospores from the mother-cell wall. Pulse-labelling studies with [3H]proline and [35S]methionine have been performed in order to detect the protein components released by synchronously growing cells at different stages of the cell cycle. When either [3H]proline or [35S]methionine were applied during the phase of cell growth, radioactive label appeared in the released macromolecules after a lag period of 40 min, whereas incorporation into the insoluble part of the cell wall was delayed only by 20 min. When applied at the end of the growth phase, e.g. 13 h after beginning of the illumination period, the radioactive amino acids were incorporated into the cell wall, but radioactive labelling of macromolecules released into the culture medium could not be detected before the zoospores were liberated from the mother-cell wall. Maximal incorporation of [3H]proline and [35S]methionine into the insoluble part of the cell wall was observed during cell division, but essentially no radioactively-labelled macromolecules were released into the culture medium during this time period. Analysis of the macromolecules, which were liberated during cell enlargement, by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis revealed distinct radioactive bands, which were differentially labelled with [3H]proline and [35S]methionine. Among the macromolecules released into the culture medium during cell growth, a component of an apparent Mr 35 000 was preferentially labelled with [3H]proline. This component was also detected after labelling with [35S]methionine, but components of an apparently higher Mr were more prominent after labelling with [35S]methionine. Macromolecules released during the cell-enlargement period of synchronously growing cultures in the presence of [3H]proline contained radioactively-labelled hydroxyproline in addition to proline. These results show that, during cell-wall growth, specific protein components are released into the culture medium and that at least one of these components contains large amounts of proline and hydroxyproline. At least some of these macromolecules seem to be constituents of the cell wall, because during pulse-chase experiments radioactively-labelled macromolecules appeared in the culture medium mainly during the time period when the specific radioactivity of the insoluble inner-cell-wall layer decreased.

Cell Cycle↗
PubMed2001

[Economic assessment of the use of tinzaparin in the treatment of acute pulmonary embolism in France].

OBJECTIVE: This pharmacoeconomic study was conducted to determine the cost of using tinzaparin, a low-molecular-weight heparin (LMWH), administered intravenously, in comparison with non-fractionated heparin (NFH) for the treatment of acute pulmonary embolism (PE) in France. METHODS: A decision algorithm indicating the therapeutic schedules and their consequences was constructed from data collected during a large-scale multicentric study and from data obtained from interviews with French physicians implicated in the treatment of pulmonary embolism. This model was used to estimate costs and to compare per patient expenditures for the two therapeutic schedules. RESULTS: Cost of instituting treatment per patient (excluding costs common to both schedules) was 202 euros for NFH and 77 euros for tinzaparin (delta 125 euros in favor of tinzaparin). Including follow-up costs for the 90 day analysis period, tizaparin reduced costs by 123 euros per patient (211 versus 334 euros for patients treated with NFH). Taking into account the sole cost of management of the two critical events that occurred during NFH or tinzaparin (8 first days) treatment, tinzaparin reduced costs by 137 euros per patient (112 euros versus 249 euros respectively). CONCLUSION: Tinzaparin reduced health care costs for pulmonary embolism due to easier administration, less complex laboratory tests, and personnel time savings. The robustness of the results was confirmed by the sensitivity analysis.

Acute Disease↗
PubMed1988

Interferon alpha-n1 (Wellferon) for refractory genital warts: efficacy and tolerance of low dose systemic therapy.

This multi-center trial compared two doses of parenterally administered interferon alpha-n1 (Wellferon) in men and women with recurrent/resistant genital warts. Patients received either 1 or 3 MU/m2 daily for 14 days, then 3 times weekly for 4 weeks; non-responders could receive an additional four weeks of treatment. A total of 107 patients were enrolled, and 102 were evaluable after six weeks of study. The principal dose comparison was in 57 women assigned alternately to the two doses. Median lesion measurements were reduced significantly from baseline at weeks 2, 4 and 6 in both groups. Statistical analysis showed no difference in response to 1 versus 3 MU/m2. The overall complete response (CR) plus partial response (PR) rate at week 6 was 69% for the two doses. Two additional groups of 21 women and 24 men were treated at the higher dose with CR plus PR rates of 75 and 50%, respectively. Week 10 disease evaluations for all groups showed 19 of 77 patients to be completely cleared. Of these 19, only one had recurrent disease at the end of the 6-month study period. Analysis of the incidence of symptomatic side effects showed a significantly higher frequency among women treated with 3 MU/m2 than among women treated with 1 MU/m2. Five dose reductions and two withdrawals for toxicity occurred, all in the high dose group. This study demonstrates that parenterally administered Wellferon produces clearance of resistant genital warts in many patients, and that rates of clearance do not appear to vary between groups receiving moderate or low dose therapy.

Adult↗
PubMed1995

Local injection of bicuculline into area 8 and area 6 of the rhesus monkey induces deficits in performance of a visual discrimination GO/NO-GO task.

While performing a symmetrically reinforced visual discrimination GO/NO-GO task, five monkeys were injected with a GABAA antagonist, bicuculline methiodide (BMI), into Brodmann's area 9, 8, 6, or 4. The task consisted of five periods: START, OFF, CUE, RESPONSE, and an inter-trial interval. The monkey was trained to make either the GO response (lever release) or the NO-GO response (continued pressing of the lever), depending on the color of the cue, during the RESPONSE period. Analysis was limited to 102 sites in which muscle convulsions of the forelimb and/or shoulder did not result from BMI injections. Errors in performance increased 10-60 min after injection into 10 of 33 sites in area 9, 9 of 25 sites in area 8, 20 of 34 sites in area 6, and 2 of 10 sites in area 4. The number of trials finished in a 120-min session decrease. Injections induced PRE-RESPONSE errors (release of the lever in either the OFF or CUE periods), GO RESPONSE errors (failure to release the lever when signaled), and NO-GO RESPONSE errors (release of the lever despite a signal not to release). The results suggest that both areas 8 and 6 are involved in correct performance of the GO/NO-GO task.

Animals↗
PubMed1998

Effects of mass screening for neuroblastoma on incidence, mortality, and survival rates in Osaka, Japan.

OBJECTIVES: To evaluate the effects of mass screening for neuroblastoma, time trends of incidence, mortality, and survival of neuroblastoma in Osaka Prefecture were analyzed. METHODS: Data for this analysis was obtained from the population-based Osaka Cancer Registry. Time trends of incidence and mortality rates were analyzed by calendar year and by birth cohort. Survival was compared between before and after the introduction of systematic screening. RESULTS: From 1970-94, 457 cases of neuroblastoma and 182 deaths from neuroblastoma were observed in Osaka. The annual age-standardized incidence rate per million children increased from 7.5 in 1970-84 to 20.5 in 1985-94, while the mortality rates did not differ between these two periods. Analysis by birth cohort showed that the incidence rate at 0 year of age per 100,000 live births increased from 2.30 in 1970-79 (unscreened) to 19.80 in 1988-89 (screening by high-performance liquid chromatography, HPLC). The incidence rate in children 1 and 2-4 years of age also increased according to the introduction of HPLC. The mortality rate in children 1-4 years of age per 100,000 live births slightly decreased from 3.87 in 1970-79 to 3.30 in 1988-89, which was presumed to be derived from the improvement in survival due to the progress in treatment. CONCLUSIONS: It is strongly suggested that mass screening for neuroblastoma causes harm because of overdiagnosis, and it has little effect on decreasing the incidence and the mortality of neuroblastoma at 1-4 years of age.

Adolescent↗
PubMed1991

Computed tomography as an adjunct in etiological analysis of hemiplegic cerebral palsy. I: Children born preterm.

Computed tomography (CT) of the brain has been used to evaluate potential etiologies of hemiplegic cerebral palsy in a population-based group of 28 children born preterm, in whom an obvious postnatal cause of hemiplegia could be excluded. The CT findings were classified according to a system so constructed to reflect the phase of brain maturation during which the insult causing hemiplegia likely had occurred. We found that 50% had evidence of periventricular leukomalacia the lesion typical of hypoxic-ischemic injury to the immature brain. However, it was not possible, neither from the CT scans nor from retrospective review of available clinical data, to date the injury more precisely to the pre- or perinatal period. Analysis of CT findings further showed that 18% of the children had cerebral maldevelopment, indicating an injury during early fetal life as the cause of hemiplegia, earlier than clinically suspected. The CT scans were normal in 25% of the children. It is concluded that the objective information obtained by studying the morphology of brain lesions using CT provides important information in addition to clinical data, and should be integrated with clinical assessment when attempts are being made to evaluate the etiology of cerebral palsy in children.

Adolescent↗
PubMedDate not supplied

The temporal relationship of psychosocial stress to cellular immunity and herpes labialis recurrences.

This study used a prospective single-subject study design and time series analysis for repeated measures data to investigate the hypothesis that variations in psychosocial stress are associated with changes in cellular immunity in a study subject with recurrent herpes labialis. A study subject who was antibody positive for HSV-1 but reported no clinical manifestations of disease served as a control. Psychosocial stress, as measured by a questionnaire; cellular immunity, as measured by the monoclonal antibodies CD4 (OKT4), which defines the helper/inducer subset of T lymphocytes; and CD8 (OKT8), which defines the suppressor/cytotoxic subset, were measured weekly over the 32-week study period. Analysis of data using bivariate time series (ARIMA) demonstrated significant inverse correlations between stress level scores and percent CD4 helper/inducer T lymphocytes in both subjects. In the study subject with recurrent herpes simplex labialis, a Mann-Whitney U statistic determined that the percent of CD4 in the early stages of a recurrence were significantly lower than the percent of CD4 at other times when blood samples were drawn. The data presented are repeated measures on two individuals with positive antibodies to the herpes simplex virus, one with recurrent herpes and one without a history of recurrent herpes. Relevance for stress research is discussed.

Adult↗
PubMed1993

Pyrrolidine dithiocarbamate inhibits NF-kappa B mobilization and TNF production in human monocytes.

The human TNF promoter contains four potential nuclear factor-kappa B (NF-kappa B)-binding sites, with the strongest binding seen for the -605 motif. Nuclear extracts from unstimulated cells of the human monocytic cell line, Mono Mac 6, contain one specific binding protein (complex II), consistent with a constitutive p50 homodimer. Stimulation of Mono Mac 6 cells with LPS will increase complex II and will strongly induce a second specific complex (complex I), which represents the p50/65 heterodimer. Treatment of Mono Mac 6 cells with pyrrolidine-dithiocarbamate (PDTC) at 300 microM will block the LPS-induced complex I almost completely and will reduce complex II to the constitutive level. Binding activity of other nuclear factors that recognize the SP-1 and c/EBP motifs of the human TNF promoter is not affected by such treatment. Northern blot analysis demonstrates that PDTC treatment will strongly reduce LPS-induced TNF transcripts. Secreted TNF protein as detected in the Wehi 164S/ActD bioassay and in a sandwich immunoassay was similarly reduced by PDTC. Kinetic analyses show that after LPS stimulation, NF-kappa B will peak at 1 h, TNF transcript prevalence at 2 h, and TNF protein at 4 h. PDTC did not shift this response to LPS to a later time, but suppressed NF-kappa B mobilization, TNF transcripts, and TNF protein over the entire 8-h observation period. Analysis of freshly isolated, LPS-stimulated blood monocytes showed a similar blockade of NF-kappa B. Furthermore, in these primary cells, induction of TNF transcripts, as determined by Northern blot analysis and by quantitative polymerase chain reaction, was prevented by PDTC as was TNF protein production. These data show that dithiocarbamates can profoundly affect cytokine expression and suggest that NF-kappa B is involved in LPS-induced TNF gene expression in human monocytes.

Binding Sites↗