Semi-automated analysis of manually reconstructed tracks of progressively motile human spermatozoa

Several alternative algorithms for computer-assisted derivationof measurements of movement characteristics from manually reconstructedtracks of progressively motile human spermatozoa were compared.Fifty tracks were reconstructed at 30 Hz from video recordingsand analysed using traditional manual methods and by four combinationsof computer algo rithms. The best algorithm set was identified(‘Video-mot.mdpt’) and the values for the curvilinear,average path and linear velocities (V_CVL, V_AVE and V_LIN respectively),the amplitude of lateral displacement of the sperm head aboutthe axis of progression (A_H) and the number of times the spermhead crossed the average path (the ‘beat/cross frequency’,BXF) obtained using it were compared to those obtained by manualanalysis. There was a considerable time saving when the computer-assistedmethod was used and the values it gave for the various movementcharacteristics showed good correspondence with those obtainedmanually. In addition, repeated data entry and analysis wasfound to be highly reproducible. When the tracks were re-plottedat 6 Hz (as used by the multiple-exposure photomicrography methodfor sperm motility analysis) insufficient information remainedin the tracks for reliable determination of anything other thanV_LIN We conclude that the Videomot.mdpt program provides reliablevalues for the movement characteristics of progressively motilehuman spermatozoa, and believe it will be of great value inthe validation of commercial systems providing automated spermmovement analysis and in laboratories which do not have accessto such costly equipment.     

Dual effects of hemicholinium-3 at central cholinergic synapses.

1. Hemicholinium-3 (HC-3) has at least two presynaptic effects at cholinergic synapses between motor acon collaterals and spinal Renshaw cells, neither of which influenced non-cholinergic excitatory synapses on dorsal horn interneurones. 2. One, which occurred independently of repetitive activity of the synapses and which resulted in increased latency of synaptic excitation, presumably involved changes in the properties of presynaptic fibres and terminals which influenced transmitter release. 3. The other, which depended on continuous repetitive synaptic activity, was consistent with depletion by HC-3 of presynaptic stores of acetylcholine (ACh).     

A contribution to sonograph image quality estimation using point spread function

In medical sonography, sonograph image quality is an essential aspect for the safety of both patient and doctor. Its evaluation therefore requires an accurate and objective method for measurement. In this regard, a number of methods are in current use. Most of these are based on tissue mimicking phantom imaging. In contrast, we have used another principle based on Point Spread Function (PSF) analysis which is a product of the measuring system we have developed. In this case, the measured sonograph scans a small metallic ball target that moves in a water bath on a specified trajectory. The Region Of Interest (ROI) of the sonogram containing the ball target picture is digitised and the amplitude of the pixels analysed. The result is the PSF from which we calculate the lateral resolution (LR). For this purpose, we use our own original software. Using this method, we have to date been able to plot LR characteristics over the scanning plane. The method allows us to differentiate separate scanning lines and even multiple focal areas for dynamic focussing systems. It can detect malfunctions in dynamic focussing, size of aperture, time gain compensation function and/or transducer element failure. The procedure itself is not as easy or as fast to use as tissue mimicking phantoms or 3D signal to noise ratio evaluation, but it provides accurate and objective numeric parameters corresponding to the quality of image at any specified point over the whole scanning area. It is also a very powerful tool when used in combination with the other methods mentioned above.     

A critical evaluation of current views regarding eye movement desensitization and reprocessing (EMDR): clarifying points of confusion.

EMDR is an active psychological treatment for PTSD that has received widely divergent reactions from the scientific and professional community. This article examines points of confusion in the published literature on EMDR, including the theoretical, empirical, and historical issues around EMDR and placebo effects, exposure procedures, the eye movement component, treatment fidelity issues, and outcome studies. It also examines historical information relevant to the scientific process and charges of "pseudoscience" regarding EMDR. We conclude that the confusion in the literature is due to (a) the lack of an empirically validated model capable of convincingly explaining the effects of the EMDR method, (b) inaccurate and selective reporting of research, (c) some poorly designed empirical studies, (d) inadequate treatment fidelity in some outcome research, and (e) multiple biased or inaccurate reviews by a relatively small group of authors. Reading the original research articles frequently helps to reduce the confusion arising from the research review literature.     

Targeted disruption of fibronectin-integrin interactions in human gingival fibroblasts by the RI protease of Porphyromonas gingivalis W50

Cell surface integrins mediate interactions between cells and their extracellular matrix and are frequently exploited by a range of bacterial pathogens to facilitate adherence and/or invasion. In this study we examined the effects of Porphyromonas gingivalis proteases on human gingival fibroblast (HGF) integrins and their fibronectin matrix. Culture supernatant from the virulent strain W50 caused considerably greater loss of the beta1 integrin subunit from HGF in vitro than did that of the beige-pigmented strain W50/BE1. Prior treatment of the W50 culture supernatant with the protease inhibitor Nalpha-p-tosyl-L-lysine chloromethyl ketone (TLCK) blocked its effects on cultured cells, indicating that this process is proteolytically mediated. Purified arginine-specific proteases from P. gingivalis W50 were able to mimic the effects of the whole-culture supernatant on loss of beta1 integrin expression. However purified RI, an alpha/beta heterodimer in which the catalytic chain is associated with an adhesin chain, was 12 times more active than RIA, the catalytic monomer, in causing loss of the alpha5beta1 integrin (fibronectin receptor) from HGF. No effect was observed on the alphaVbeta3 integrin (vitronectin receptor). The sites of action of RI and RIA were investigated in cells exposed to proteases pretreated with TLCK to inactivate the catalytic component. Use of both monoclonal antibody 1A1, which recognizes only the adhesin chain of RI, and a rabbit antibody against P. gingivalis whole cells indicated localization of RI on the fibroblasts in a clear, linear pattern typical of that seen with fibronectin and alpha5beta1 integrin. Exact colocalization of RI with fibronectin and its alpha5beta1 receptor was confirmed by double labeling and multiple-exposure photomicroscopy. In contrast, RIA bound to fibroblasts in a weak, patchy manner, showing only fine linear or granular staining. It is concluded that the adhesin component of RI targets the P. gingivalis arginine-protease to sites of fibronectin deposition on HGF, contributing to the rapid loss of both fibronectin and its main alpha5beta1 integrin receptor. Given the importance of integrin-ligand interactions in fibroblast function, their targeted disruption by RI may represent a novel mechanism of damage in periodontal disease.     

Extensive transmission of Mycobacterium tuberculosis from a child

BACKGROUND AND METHODS: Young children rarely transmit tuberculosis. In July 1998, infectious tuberculosis was identified in a nine-year-old boy in North Dakota who was screened because extrapulmonary tuberculosis had been diagnosed in his female guardian. The child, who had come from the Republic of the Marshall Islands in 1996, had bilateral cavitary tuberculosis. Because he was the only known possible source for his female guardian's tuberculosis, an investigation of the child's contacts was undertaken. We identified family, school, day-care, and other social contacts and notified these people of their exposure. We asked the contacts to complete a questionnaire and performed tuberculin skin tests. RESULTS: Of the 276 contacts of the child whom we tested, 56 (20 percent) had a positive tuberculin skin test (induration of at least 10 mm), including 3 of the child's 4 household members, 16 of his 24 classroom contacts, 10 of 32 school-bus riders, and 9 of 61 day-care contacts. A total of 118 persons received preventive therapy, including 56 young children who were prescribed preventive therapy until skin tests performed at least 12 weeks after exposure were negative. The one additional case identified was in the twin brother of the nine-year-old patient. The twin was not considered infectious on the basis of a sputum smear that was negative on microscopical examination. CONCLUSIONS: This investigation showed that a young child can transmit Mycobacterium tuberculosis to a large number of contacts. Children with tuberculosis, especially cavitary or laryngeal tuberculosis, should be considered potentially infectious, and screening of their contacts for infection with M. tuberculosis or active tuberculosis may be required.     

Excitatory amino acids mediate responses elicited in vitro by stimulation of cortical afferents to reticularis thalami neurons of the rat

The effects of the excitatory amino acids on the nucleus reticularis thalami were examined by intracellular recordings from rat thalamic slices. Non-N-methyl-D-aspartate receptor agonists and glutamate induced a membrane depolarization and a reduction in input resistance, while N-methyl-D-aspartate and aspartate induced a prolonged discharge, which in some neurons took the form of a burst firing associated with an apparent increase in membrane input resistance. Both the N-methyl-D-aspartate and the aspartate effects were blocked by D-2-amino-5-phosphonovalerate, while the effects of glutamate, kainate and quisqualate were not. The excitatory postsynaptic potential evoked by corticothalamic fiber stimulation shows two components: an early, short-lasting, 2-amino-5-phosphonovalerate-insensitive portion, and a late, 2-amino-5-phosphonovalerate-sensitive decay phase. It is suggested that glutamate acts in nucleus reticularis thalami cells preferentially on the non-N-methyl-D-aspartate receptors, while aspartate shows an N-methyl-D-aspartate-like effect. The two excitatory amino acids glutamate and aspartate play a determinant role in the modulation of thalamic activity driven by corticothalamic projection.     

Complete Nucleotide Sequence Analysis of a Western Pacific Dengue-1 Virus Strain

We have determined the complete nucleotide sequence and the deduced amino acid polypeptide sequence of the genome of a dengue-1 (DEN-1) virus strain isolated from a patient on Nauru in the Western Pacific in 1974 (West Pac 74). The complete genome is 10,735 nucleotides in length and contains a single long open reading frame of 10,176 nucleotides encoding a polyprotein of 3392 amino acids. When compared to DEN-1 Singapore S275/90, the nucleotide and amino acid sequence homology are 94% and 97.8%, respectively. This revised version was published online in July 2006 with corrections to the Cover Date.