Detection of an anatoxin-a(s)-like anticholinesterase in natural blooms and cultures of cyanobacteria/blue-green algae from Danish lakes and in the stomach contents of poisoned birds

Ten natural bloom samples of cyanobacteria from the Danish lakes Knud s? (5), Ravn s? (4), and Salten Langs? (1) collected during 1993-1995 were assayed for toxicity by mouse bioassay, for acetylcholinesterase inhibiting activity by a colorimetric method, and for microcystins by enzyme-linked immunosorbent assay. In the mouse bioassay, seven samples were neurotoxic, two were non-toxic and one gave a protracted toxic response. One of the non-toxic and the single protracted toxic sample both contained anticholinesterase activity equivalent to 4 micrograms anatoxin-a(s) g-1. The neurotoxic samples contained equivalents to 20-3300 micrograms anatoxin-a(s) g-1. The highest anticholinesterase activities (equivalent to 2300 and 3300 micrograms anatoxin-a(s) g-1, respectively) were found in samples collected from Lake Knud s? in connection with bird-kills in 1993 and 1994. Small amounts of microcystins (0.1-0.9 microgram g-1) were detected in all samples but one. All Lake Knud s? and Lake Ravn s? samples were dominated by Anabaena lemmermannii, and the Lake Salten Langs? sample by several species of Anabaena. Gel filtration profiles indicated similarity between the toxic component from the Lake Knud s? 1994 bloom with registered bird-kills and anatoxin-a(s) isolated from Anabaena flos-aquae NRC-525-17. Anticholinesterase-producing cultures of A. lemmermannii were isolated from the Lake Knud s? 1993 bloom. These laboratory cultures produced anatoxin-a(s) equivalents of 29-743 micrograms g-1. Other cultures of A. lemmermannii isolated from Lake Knud s? and Lake Ravn s? were hepatotoxic or non-toxic. Dead birds collected from Lake Knud s? during the neurotoxic 1993 Anabaena bloom possibly died from cyanobacterial toxicosis. The stomach contents contained colonies and single trichomes of Anabaena, and anticholinesterase activities equivalent to 2.1-89.7 micrograms anatoxin-a(s) kg-1 body weight and microcystins (53-95 ng kg-1) were also detected.     

Ascorbic acid is a stimulatory cofactor for mitochondrial glycerol-3-phosphate dehydrogenase

We describe in this report a sensitive and direct method for the analysis of tamoxifen (TAM) in microsamples of plasma. The drug and internal standard (quinine bisulfate, I.S.) were separated on a 10-microm particle, 10 cm X 8 mm CN cartridge in conjunction with a radial compression system. The mobile phase was a mixture of 0.1 M sodium acetate in 0.001 M tetrabutylammonium phosphate solution (pH 6) and methanol (30:70, v/v) at a flow-rate of 4 ml/min. After addition of I.S. and o-phosphoric acid in acetonitrile (0.6 M) to the plasma (30 microl), the mixture was placed in an ultraviolet shortwave transluminator for 2 min prior to injection into the chromatograph. The compounds were detected in the effluent fluorometrically at excitation and emission wavelengths of 258 and 378 nm, respectively. Under these conditions, no interference in the assay from any endogenous substance or other concurrently used drugs was observed and the retention times of I.S. and TAM were 4.4 and 10.15 min, respectively. The concentration of TAM in plasma was linearly (r>0.9983) related to the peak height ratio (TAM/I.S.) in the range 0.01-2.0 microg ml(-1) and C.V. at 0.075, 0.4 and 1.2 microg ml(-1) was < or = 4.96%. We are currently using this assay for monitoring TAM in plasma and investigating its pharmacokinetics in cancer patients receiving cytotoxic drugs in addition to TAM as a multi-drug resistance modifier.     

NK cell proliferation and inflammation

Considerable progress has been made in recent years in understanding the biology of NK cells. NK cells are no longer 'null cells', but express an array of functionally important molecules with which they mediate and regulate their cytolytic activity and the cytokines they secrete. Activation and proliferation of NK cells in influenced by cytokines produced by activated monocytes (IL-15, IL-12, IL-10) and activated T cells (IL-2). This paper reviews the phenotype and effector functions of NK cells, their tissue distribution, and evidence that NK cells proliferate in vivo as part of productive or pathologic consequences of immunity.     

Administration of first-aid and prevention of injuries in Kenyan soccer

Appropriate scientific training demands the cooperation of the coach, player, doctor and physiotherapist. The training process involves the medical examination before a competitive season, the diagnosis and treatment of injuries when they occur, the prevention of injuries and the rehabilitation of the injured back into competitive play. It is in these respects that a coach requires the services of a doctor and/or a physiotherapist on the technical bench. This study therefore attempted to establish whether the soccer coaches were benefiting from the professional support of the doctors. The study sought the views of the Kenyan soccer coaches on the administration of first-aid and the preventive measures against injuries. Specifically, the study attempted to find out whether the Kenyan soccer clubs employed team doctors, whether the coaches had adequate knowledge in first-aid, the injury prevention measures that the coaches enforced, etc. A questionnaire was administered to 42 individual soccer coaches who were attending an advanced Confederation Africaine de Football (CAF) coaching course in Nairobi. Thirty-three (78.57%) were used in the study. The participants were drawn from all the provinces of Kenya except North Eastern. The findings were that 13 (39.4%) of the clubs represented did not have a qualified medical attendant; that in the absence of a team doctor, it is the coach who mostly administered first-aid (42.4%); that out of all the coaches only 15 (45.5%) were well versed in first-aid procedures, among others.(ABSTRACT TRUNCATED AT 250 WORDS)     

Sensitivity analysis of initial value problems with mixed odes and algebraic equations

An efficient method is described for sensitivity analysis of nonlinear initial value problems, which may include algebraic equations as well as ordinary differential equations.The linearity of the sensitivity equations is utilized to solve them directly via the local Jacobian of the state equations. The method is implemented with the implicit integrator DASSL and is demonstrated on a stiff industrial reaction model.     

Increased synthetic peptide specificity of tissue-CSF bound anti-MBP in multiple sclerosis

Free and bound hydrosoluble protein extracts were prepared from four anatomical areas of a multiple sclerosis (MS) cerebrum and from corresponding anatomical areas of a normal (non-MS) control. Increased levels of IgG and anti-myelin basic protein antibodies (anti-MBP) were detected in all MS samples and they were undetectable in the controls. IgG and anti-MBP from free (unbound) hydrosoluble protein extracts are defined as free IgG and free anti-MBP while IgG and anti-MBP from tissue bound protein extracts are defined as bound IgG and bound anti-MBP. IgG was purified from free protein extracts by protein G Sepharose affinity chromatography and anti-MBP was further isolated from purified IgG by antigen specific (MBP) Sepharose affinity chromatography. Free and bound anti-MBP were reacted with 20 synthetic peptides of human MBP prepared by the Fmoc method. Free anti-MBP, whether in the context of whole protein extracts, or as purified IgG or as purified antibody was completely neutralized by peptides #12, #15, #56 and #56* containing overall residues 75-106, partially neutralized by peptides #27, #16 and #21 containing overall residues 61-83 and did not react with the remaining 13 peptides. Tissue bound anti-MBP was completely neutralized only by peptides #12, #15, #56 and #56* (overall residues 75-106) and showed no reactivity towards the remaining 16 peptides including peptides #27, #16 and #21. Synthetic peptide specificity of free anti-MBP purified from MS cerebrum was identical to previously reported specificity of free anti-MBP from MS cerebrospinal fluid (CSF), while tissue bound anti-MBP, as well as bound anti-MBP from CSF had a more restricted synthetic peptide specificity than free anti-MBP. This suggests that the most likely epitope of anti-MBP is located between residues 84 and 95 of human MBP just proximal to the tri-proline sequence (99-101).     

Cortical implants for the blind

Arrays of stimulating electrodes can be placed in the brain itself, in the visual cortex, to bring vision to the profoundly blind. However, when designing such devices one needs to take into consideration the fact that the visual pathway maps images onto cortical structures in a complex and unpredictable way