Picoplankton dynamics and picoeukaryote diversity in a hyper-eutrophic subtropical lagoon.

Picoplankton (cells with a diameter of 0.2-3.0 µm) is the dominant contributor to both primary production and biomass in the ocean. Most of the previous studies on picoplankton have been conducted in the oligotrophic open sea with few in the eutrophic area. In this study, we investigated the dynamics of different groups of picoplankton and the diversity of picoeukaryote (based on 18S rDNA) in a hyper-eutrophic marine coastal lagoon. The results indicated that temperature and phosphate concentration were most responsible for the dynamics of different picoplankton groups. Examination of 135 clones revealed 27 different Denaturing Gradient Gel Electrophoresis (DGGE) patterns. At least 7 high-level taxonomic groups of picoeukaryote were recorded. The picoeukaryotic diversities included Alveolates, Stramenopiles, Haptophyceae, and Viridiplantae, with Stramenopiles being the most diverse group. Overall the results of this study indicated that picoplankton diversity was low relative to studies conducted in more oligotrophic waters.

The effect of Chinese herbal medicines (CHM) on menopausal symptoms compared to hormone replacement therapy (HRT) and placebo.

OBJECTIVE: To evaluate within the Traditional Chinese Medicine (TCM) setting, the effect of CHM-formulae on menopausal symptoms. DESIGN: A double-blind and double-dummy randomised placebo-controlled trial. METHOD: Between February and June 2002 and June and October 2004, 31 peri- and postmenopausal Dutch women were recruited to complete 12 weeks of treatment with either CHM formulae (n=10), HRT (n=11) or placebo (n=10) medications plus 4 weeks of non-treatment follow-up observation. Hereby a double-dummy setting is applied, i.e. the medications and placebo's taken were both capsules and liquid extracts. Appropriate to the TCM setting, the CHM-prescriptions could be adjusted according to the symptoms and signs of the individuals for that moment. MAIN OUTCOME MEASURES: The primary end-point was the reduction in frequency of vasomotor symptoms (hot flushes and night sweats). Secondary end-point were the improvements measured in quality of life questionnaire SF-36 and other symptoms and signs related to the peri- and postmenopausal period. RESULTS: Placebo had a score of 30%. Compared to Placebo, on average CHM is 29% significantly (p<0.05) more effective in reducing the amount of hot flushes, while HRT is almost 50%. Although quantitatively there was a significant difference in the reduction of hot flushes between groups, qualitatively there was no overall improvement. CONCLUSIONS: This pilot study proved clearly that CHM could help women with their menopausal problems. The chosen trial methodology with its TCM differential diagnosis after orthodox medicine diagnosis is fully compatible with TCM practice and hence acceptable for western and Chinese medical practitioners. For a place in the western health care system, we need to conduct a larger trial with a more menopause specific questionnaire.

Towards establishing comprehensive databases of cellular proteins from transformed human epithelial amnion cells (AMA) and normal peripheral blood mononuclear cells.

Databases of protein information derived from the analysis of two-dimensional gels have been established from transformed human amnion cells (AMA) and peripheral blood mononuclear cells (PBMCs). A total of 1781 [35S]methionine-labeled AMA proteins (1274 IEF, 537 NEPHGE) and a total of 1311 proteins from PBMC (948 IEF, 363 NEPHGE) were resolved and recorded using computerized (PDQ-SCAN and PDQUEST softwares) two-dimensional gel electrophoresis. AMA and PBMC proteins (total, 454: 301 IEF, 153 NEPHGE) were matched both manually and by the computer. Information entered in the AMA database (in most cases for some major proteins) includes: molecular weight, protein name, HeLa protein catalogue number, mouse protein catalogue number, nuclear proteins, phosphorylated proteins, distribution of proteins in Triton X-100 supernatants and cytoskeletons, proliferation- and transformation-sensitive proteins, cell cycle-specific proteins, mitochondrial proteins, proteins matched in normal human embryonal lung MRC-5 fibroblasts and PBMC cells, heat shock proteins, proteins affected by interferons, cytoskeletal proteins, and the presence of antibody against protein in human sera. Additional information has been entered for the cell cycle-regulated and DNA replication protein cyclin (PCNA). Information entered in the PBMC database includes molecular weight and potential markers for sorted populations of lymphocyte subtypes. For those proteins that have been matched to AMA proteins, information contained in some entries may be transferred from the AMA database.

Comprehensive, human cellular protein databases and their implication for the study of genome organization and function.

Comprehensive, computerized databases of cellular protein information derived from the analysis of two-dimensional gels, together with recently developed techniques to microsequence proteins offer a new dimension to the study of genome organization and function. In particular, human protein databases provide an ideal framework in which to focus the human genome sequencing effort.

Occupational exposures and knowledge of universal precautions among medical students.

PURPOSE: To examine the relationship between occupational exposures and knowledge of universal precautions among medical students. METHOD: Graduating medical students were given a survey regarding occupational exposures suffered during their clinical rotations. The survey also tested students' knowledge of universal precautions by asking them to indicate what combination of gloves, mask, and eyeshields should be worn to satisfy universal precautions for ten common procedures. RESULTS: At a seminar one week before graduation, 45 senior medical students were given the questionnaire. The response rate was 100% 84% of the surveyed students suffered at least one occupational exposure during their clinical training. Of those who had an exposure, 42% reported at least once to an exposure center. The mean percentage of correct answers on the protective equipment questionnaire was 71%. No correlation between number of exposures and score on the protective equipment questionnaire was found (r = 0.0). CONCLUSION: Occupational exposures to blood are common among medical students. Few students report to exposure centers. Knowledge of universal precautions may not correlate with reduced risk of occupational exposures among medical students.

Kinetics of polychlorinated biphenyl partitioning to marine Chrysophyte Isochrysis galbana.

This study focused on the uptake kinetics of polychlorinated biphenyl (PCB) congeners by the Chrysophyte, Isochrysis galbana. A gas-purging experimental system was used to maintain constant dissolved PCB concentrations. Three phases of absorption were observed: first, a rapid absorption phase within the first 15min, second, a first order process reaching the maximum concentration within 48h of exposure, and third, a plateau phase as yet to be determined with very slight increases in concentration. In this study, the percentage of the maximum concentration reached within the first phase varied from 10% to 67%, depending on the size of the PCB (as determined by molecular weight and total surface area), whereas the uptake rate (k(u)) during the second phase was more comparable across different PCBs. In addition, for the first phase, the bioconcentration factor (BCF) of PCBs deviated from its expected relationship with hydrophobicity, as determined by K(ow), and was instead related to the molecular structure of the compound.

The effects of radiofrequency fields on cell proliferation are non-thermal.

The number of reports on the effects induced by radiofrequency (RF) electromagnetic fields and microwave (MW) radiation in various cellular systems is still increasing. Until now no satisfactory mechanism has been proposed to explain the biological effects of these fields. One of the current theories is that heat generation by RF/MW is the cause, in spite of the fact that a great number of studies under isothermal conditions have reported significant cellular changes after exposure to RF/MW. Therefore, this study was undertaken to investigate which effect MW radiation from these fields in combination with a significant change of temperature could have on cell proliferation. The experiments were performed on the same cell line, and with the same exposure system as in a previous work [S. Kwee, P. Raskmark, Changes in cell proliferation due to environmental non-ionizing radiation: 2. Microwave radiation, Bioelectrochem. Bioenerg., 44 (1998), pp. 251-255]. The field was generated by signal simulation of the Global System for Mobile communications (GSM) of 960 MHz. Cell cultures, growing in microtiter plates, were exposed in a specially constructed chamber, a Transverse Electromagnetic (TEM) cell. The Specific Absorption Rate (SAR) value for each cell well was calculated for this exposure system. However, in this study the cells were exposed to the field at a higher or lower temperature than the temperature in the field-free incubator i.e., the temperature in the TEM cell was either 39 or 35 +/- 0.1 degrees C. The corresponding sham experiments were performed under exactly the same experimental conditions. The results showed that there was a significant change in cell proliferation in the exposed cells in comparison to the non-exposed (control) cells at both temperatures. On the other hand, no significant change in proliferation rate was found in the sham-exposed cells at both temperatures. This shows that biological effects due to RF/MW cannot be attributed only to a change of temperature. Since the RF/MW induced changes were of the same order of magnitude at both temperatures and also comparable to our previous results under isothermal conditions at 37 degrees C, cellular stress caused by electromagnetic fields could initiate the changes in cell cycle reaction rates. It is widely accepted that certain classes of heat-shock proteins are involved in these stress reactions.

Mode of interaction of polyoxyethyleneglycol detergents with membrane proteins.

Binding of dodecyloctaethyleneglycol monoether (C12E3) and purified Triton X-100 to various integral membrane proteins was studied by chromatographic procedures. Binding capacity decreased in the following order: bovine rhodopsin greater than photochemical reaction center greater than sarcoplasmic reticulum Ca2+-ATPase. The detergents were bound in different amounts to the proteins and less than corresponding to the aggregation number of the pure micelles. Appreciable binding of C12E8 to Ca2+-ATPase was observed far below the critical micelle concentration, consistent with interaction of the membrane protein with non-micellar detergent. Model calculations indicate that the detergents cannot combine with the membrane proteins, forming an oblate ring similar to that of pure detergent micelles, such as has been previously proposed for e.g. cytochrome b5 [Robinson and Tanford (1975) Biochemistry, 14, 365-378]. Other arrangements (prolate and monolayer rings), in which all detergent molecules are in contact with the protein, are considered as alternatives for covering the hydrophobic surface of the membrane protein with a continuous layer of detergent.

Positron emission tomography study of voluntary saccadic eye movements and spatial working memory.

1. The purpose of this study is to define the cortical regions that subserve voluntary saccadic eye movements and spatial working memory in humans. 2. Regional cerebral blood flow (rCBF) during performance of oculomotor tasks was measured with [15O]-H2O positron emission tomography (PET). Eleven well-trained, healthy young adults performed the following tasks: visual fixation, visually guided saccades, antisaccades (a task in which subjects made saccades away from rather than toward peripheral targets), and either an oculomotor delayed response (ODR, a task requiring memory-guided saccades after a delay period) or a conditional antisaccade task (a task in which the color of the peripheral target determined whether a saccade toward or away from the target was required). An additional six subjects performed a sequential hand movement task to compare localization of hand-related motor cortex and the frontal eye fields (FEFs) and of the hand- and eye-movement-related regions of the supplementary motor area (SMA). 3. Friston's statistical parametric mapping (SPM) method was used to identify significant changes in rCBF associated with task performance. Because SPM does not take advantage of the anatomic information available in magnetic resonance (MR) scans, each subject's PET scan was registered to that individual's MR scan, after which all PET and MR studies were transformed to conform to a standard reference MR image set. Subtraction images were visually inspected while overlayed on the reference MR scan to which PET images had been aligned, in order to confirm anatomic localization of significant rCBF changes. 4. Compared with visual fixation, performing visually guided saccades led to a significant bilateral activation in FEF, cerebellum, striate cortex, and posterior temporal cortex. Right posterior thalamus activation was also observed. 5. The visually guided saccade task served as the comparison task for the ODR, antisaccade, and conditional antisaccade tasks for identification of task-related changes in rCBF beyond those associated with saccade execution. Performance on the ODR task was associated with a bilateral increase of rCBF in FEFs, SMA, dorsolateral prefrontal cortex (DLPFC), and posterior parietal cortex. The cortical regions of increased regional blood flow during the ODR task also showed increased rCBF during the antisaccade task; however, FEF and SMA activations were significant only in the right hemisphere. These findings closely parallel those of single-cell recording studies with behaving monkeys in indicating that FEF, DLPFC, SMA, and posterior parietal cortex perform computational activity for voluntary purposive saccades. 6. Comparison of PET scans obtained during performance of eye movement and hand movement tasks indicated that peak activations in FEF were located approximately 2 cm lateral and 1 cm anterior to those of hand-related motor cortex. The oculomotor area of SMA, the supplementary eye field (SEF), was located approximately 7-8 mm anterior and superior to the hand-related area of SMA. 7. During performance of antisaccade and ODR tasks, rCBF was significantly lower in ventromedial prefrontal cortex (PFC), along the rectus gyrus, and in ventral anterior cingulate cortex than during the visually guided saccade and fixation tasks. During the antisaccade task, the ventral region of lower rCBF involved medial structures including left ventral striatum and bilateral medial temporal-limbic cortex. During the ODR task, the ventral aspect of the region of lower rCBF extended laterally, rather than medially, to include the temporal poles. The lower blood flow observed in ventromedial PFC during both the antisaccade and ODR tasks, relative to the visually guided saccade and fixation tasks, suggests that modulation of output from ventromedial PFC to limbic cortex and the striatum may play a role in the voluntary control of saccadic eye movements, possibly in the suppression of responses that would interrupt

Computerized, comprehensive databases of cellular and secreted proteins from normal human embryonic lung MRC-5 fibroblasts: identification of transformation and/or proliferation sensitive proteins.

Databases of protein information from human embryonal lung fibroblasts (MRC-5) have been established using computer analyzed two-dimensional gel electrophoresis. One thousand four hundred and eighty-two cellular proteins (1060 with isoelectric focusing and 422 with nonequilibrium pH gradient electrophoresis, in the first dimension) ranging in molecular mass between 8 and 234 kDa were separated and numbered. Information entered in the database (in most cases for major proteins) includes: protein name, HeLa protein catalog number, mouse protein catalog number, proteins matched in transformed human epithelial amnion cells (AMA) and peripheral blood mononuclear cells (PBMC), transformation and/or proliferation sensitive proteins, synthesis in quiescent cells, cell cycle regulated proteins, mitochondrial and heat shock proteins, cytoskeletal proteins and proteins whose synthesis is affected by interferons. Additional information entered for a few transformation-sensitive proteins that have been selected for future studies includes levels of synthesis and amounts in fetal human tissues. A total of four hundred and seventy-six [35S]methionine labeled polypeptides (258 isoelectric focusing; 218, nonequilibrium pH gradient electrophoresis) secreted by MRC-5 fibroblasts were separated and recorded (J. E. Celis et al., Leukemia 1987, 1, 707-717). Information entered in this database includes molecular weight and transformation sensitive proteins. These databases, as well as those of epithelial and lymphoid cell proteins (J. E. Celis et al., Leukemia 1988, 9, 561-601), represent the initial stages of a systematic effort to establish comprehensive databases of human protein information. In the long run, these databases are expected to offer a useful framework in which to focus the human genome sequencing effort.