Three-dimensional microscale hanging drop arrays with geometric control for drug screening and live tissue imaging.

Existing three-dimensional (3D) culture techniques are limited by trade-offs between throughput, capacity for high-resolution imaging in living state, and geometric control. Here, we introduce a modular microscale hanging drop culture where simple design elements allow high replicates for drug screening, direct on-chip real-time or high-resolution confocal microscopy, and geometric control in 3D. Thousands of spheroids can be formed on our microchip in a single step and without any selective pressure from specific matrices. Microchip cultures from human LN229 glioblastoma and patient-derived mouse xenograft cells retained genomic alterations of originating tumors based on mate pair sequencing. We measured response to drugs over time with real-time microscopy on-chip. Last, by engineering droplets to form predetermined geometric shapes, we were able to manipulate the geometry of cultured cell masses. These outcomes can enable broad applications in advancing personalized medicine for cancer and drug discovery, tissue engineering, and stem cell research.

Mindfulness-based stress reduction alters brain activity for breast cancer survivors with chronic neuropathic pain: preliminary evidence from resting-state fMRI.

PURPOSE: Breast cancer continues to be the most commonly diagnosed cancer among Canadian women, with as many as 25-60% of women suffering from chronic neuropathic pain (CNP) as a pervasive consequence of treatment. While pharmacological interventions have shown limited efficacy for the management of CNP to date, psychological interventions, such as mindfulness-based stress reduction (MBSR), may be a promising alterative for improving pain-related problems. The purpose of this study was to use brain imaging methods to investigate this potential. METHODS: Resting-state fMRI was used in female breast cancer survivors with CNP before and after an 8-week MBSR course (n = 13) and compared with a waitlist control group (n = 10). RESULTS: Focusing on the default mode network, the most significant results show greater posterior cingulate connectivity with medial prefrontal regions post-MBSR intervention. Moreover, this change in connectivity correlated with reduced pain severity for the MBSR group. CONCLUSIONS: These results provide empirical evidence of a change in the brain following MBSR intervention associated with changes in the subjective experience of pain. IMPLICATIONS FOR CANCER SURVIVORS: This study gives hope for a non-invasive method of easing the struggle of CNP in women following breast cancer treatment.

Preindustrial 14CH4 indicates greater anthropogenic fossil CH4 emissions.

Atmospheric methane (CH4) is a potent greenhouse gas, and its mole fraction has more than doubled since the preindustrial era1. Fossil fuel extraction and use are among the largest anthropogenic sources of CH4 emissions, but the precise magnitude of these contributions is a subject of debate2,3. Carbon-14 in CH4 (14CH4) can be used to distinguish between fossil (14C-free) CH4 emissions and contemporaneous biogenic sources; however, poorly constrained direct 14CH4 emissions from nuclear reactors have complicated this approach since the middle of the 20th century4,5. Moreover, the partitioning of total fossil CH4 emissions (presently 172 to 195 teragrams CH4 per year)2,3 between anthropogenic and natural geological sources (such as seeps and mud volcanoes) is under debate; emission inventories suggest that the latter account for about 40 to 60 teragrams CH4 per year6,7. Geological emissions were less than 15.4 teragrams CH4 per year at the end of the Pleistocene, about 11,600 years ago8, but that period is an imperfect analogue for present-day emissions owing to the large terrestrial ice sheet cover, lower sea level and extensive permafrost. Here we use preindustrial-era ice core 14CH4 measurements to show that natural geological CH4 emissions to the atmosphere were about 1.6 teragrams CH4 per year, with a maximum of 5.4 teragrams CH4 per year (95 per cent confidence limit)-an order of magnitude lower than the currently used estimates. This result indicates that anthropogenic fossil CH4 emissions are underestimated by about 38 to 58 teragrams CH4 per year, or about 25 to 40 per cent of recent estimates. Our record highlights the human impact on the atmosphere and climate, provides a firm target for inventories of the global CH4 budget, and will help to inform strategies for targeted emission reductions9,10.

Mechanical and release behaviour of theophylline from matrix tablets containing psyllium powder in combination with grewia polysaccharides.

This study was aimed at investigating the effect of grewia polysaccharides on the mechanical and release properties of tablet matrices containing binary mixtures of the polysaccharide with psyllium. Two grades of grewia polysaccharides (GG and GDS) were extracted and binary mixtures of the polysaccharides with psyllium were formulated into tablet matrices containing theophylline as the model drug. The true, bulk and tapped densities, Carr's compressibility index of the powders and binary composites were determined before tablet compression. Tablet properties (hardness, porosity, and drug release from the matrices) were investigated. The dissolution test was carried out in 0.1 M HCl (pH 1.2) and phosphate buffer (pH 6.8). The results show that GG and GDS produced tablets with good mechanical strength (108.33 N and 95.70 N, respectively) while psyllium produced softer tablets (7.13 N). The combination of psyllium and grewia polysaccharides in the matrices resulted in a significant increase in the mechanical strength of the matrices when compared to matrices containing psyllium alone as the matrix former. The results also showed that GG and GDS reduced the dissolution rate and effectively eliminated the burst release of theophylline from the psyllium matrices at both pHs. The matrices of GG or GDS and the binary mixtures conform to non-Fickian anomalous diffusion with n > 0.45. When overcoming the burst release of drug from matrices such as psyllium, grewia polysaccharides may provide an effective reduction and a more sustained drug release from such matrices.

Starch-free grewia gum matrices: Compaction, swelling, erosion and drug release behaviour.

Polysaccharides are suitable for application as hydrophilic matrices because of their ability to hydrate and swell upon contact with fluids, forming a gel layer which controls drug release. When extracted from plants, polysaccharides often contain significant quantities of starch that impacts upon their functional properties. This study aimed to evaluate differences in swelling, erosion and drug release from matrix tablets prepared from grewia gum (GG) and starch-free grewia gum (GDS) extracted from the stems of Grewia mollis. HPMC was used as a control polymer with theophylline as a model drug. Swelling, erosion, and in-vitro release were performed in deionized water, pH 1.2 and pH 6.8 media. The Vergnaud and Krosmeyer-Peppas model were used for swelling and drug release kinetics, respectively. However, linear regression technique was used to determine the erosion rate. GDS compacts were significantly harder than the native GG and HPMC compacts. GDS matrices exhibited the fastest erosion and drug release in deionised water and phosphate buffer compared with the GG and HPMC. At pH 1.2, GDS exhibited greater swelling than erosion, and drug release was similar to GG and HPMC. This highlights the potential of GDS as a matrix for controlled release similar to HPMC and GG at pH 1.2 but with a more rapid release at pH 6.8. GDS may have wider application in reinforcing compacts with relatively low mechanical strength.

Technical note: Comparison of radial immunodiffusion and ELISA for quantification of bovine immunoglobulin G in colostrum and plasma.

Historically, radial immunodiffusion (RID) has been the only method that directly measures IgG; however, recent studies have reported IgG concentrations in colostrum, milk, and plasma as measured using an ELISA. To our knowledge no comparison between RID and ELISA methods has been made for bovine colostrum or plasma. The objective of this study was to compare IgG concentrations measured by both methods in samples of bovine colostrum before and after heat treatment and bovine plasma. Concentration of IgG was quantified using a commercially available RID kit and a modified ELISA. Samples of bovine colostrum and plasma were collected from individual animals and colostrum was tested before and after heat treatment at 60°C for 30 min. All samples were tested using both methods. Pearson correlation coefficients were determined for RID and ELISA values from unheated colostrum, heat-treated colostrum, and plasma samples. Mixed models were used to determine the effect of assay on IgG measurement in colostrum and plasma and effect of heat treatment on IgG concentration in colostrum. A weak correlation was found between ELISA and RID results in plasma and unheated colostrum. Concentration of IgG was significantly lower in all sample types when measured by ELISA compared to RID. Thus, direct comparison of ELISA and RID results is not recommended. Colostrum IgG concentration significantly decreased after heat treatment as measured by ELISA, but means were not different when measured by RID. Correlation plots between colostrum values measured before and after heat treatment indicated changes in the colostrum protein matrix due to heat affected RID and ELISA assays differently. This investigation compared RID and ELISA results, but no conclusions could be drawn as to the accuracy of either assay.

Venous thromboembolism in patients receiving perioperative chemotherapy for esophagogastric cancer.

The association between venous thromboembolism and chemotherapy for esophagogastric cancer is well known in patients treated with palliative intent. Whether this risk extends to the neoadjuvant and perioperative setting is unclear. A retrospective interrogation of databases of patients receiving perioperative chemotherapy for potentially curative intent at the Leicester (2006-2011) and Nottingham (2004-2011) esophagogastric cancer centers was performed. Thromboembolic events were diagnosed in 48 of 384 patients (12.5%), 21 (5.5%) at presentation, 12 (3%) during neoadjuvant chemotherapy, and 15 (3.9%) in the postoperative period. There were no deaths from thromboembolic disease. By site these comprised catheter-related axillary vein thrombosis in 7 patients, deep venous thrombosis in 12 patients, and pulmonary embolism in 29 patients. Twenty-five of the 29 pulmonary emboli were incidental findings on staging computed tomography imaging. Combination chemotherapy with epirubicin, cisplatin, and capecitabine appeared to carry the greatest risk for the development of thromboembolism. Seven of the 12 patients (58%) who developed thromboembolism during neoadjuvant chemotherapy did not proceed to surgery because of deterioration in performance status. Preoperative thromboembolic disease resulted in a significant increase in the interval between chemotherapy and surgery, but did not influence either length of hospital stay or survival. Venous thromboembolism will develop in 12.5% of patients treated with potentially curative intent. This adverse event can occur at any time during the patient journey. In contrast to the commonly held view, this did not translate into a poorer prognosis.

Valproic acid enhances microglial phagocytosis of amyloid-beta(1-42).

Alzheimer's disease (AD) is a prevalent neurodegenerative disorder manifested by memory loss, confusion and changes in mood. A principal pathology of this debilitating disorder is extracellular deposits of amyloid-beta (Abeta) protein. The "amyloid hypothesis" postulates that a build-up of Abeta protein is responsible for neuronal loss and the ensuing symptoms of AD. One possible mechanism of Abeta clearance, and hence AD therapy, is phagocytosis of Abeta protein by microglial cells. Microglia are the brain's resident immune cells and phagocytosis is one of their innate functions. We are interested in identifying molecules that augment microglial-mediated phagocytosis of Abeta protein. We used the rodent BV-2 microglial cell line which readily phagocytose fluorescent latex beads and synthetic Abeta(1-42) peptide. BV-2 cells treated with the neuroactive drug valproic acid (VPA) showed greatly enhanced phagocytic activity for both latex beads and Abeta. VPA also reduced microglial viability by inducing apoptosis, as previously reported. The relevance of these in vitro results to the treatment of AD is unclear but further investigation into the effects of VPA on the clearance of Abeta through enhanced microglial phagocytosis is warranted.

Response of cytoplasmic pH to anoxia in plant tissues with altered activities of fermentation enzymes: application of methyl phosphonate as an NMR pH probe.

BACKGROUND AND AIMS: Acidification of the cytoplasm is a commonly observed response to oxygen deprivation in plant tissues that are intolerant of anoxia. The response was monitored in plant tissues with altered levels of lactate dehydrogenase (LDH) and pyruvate decarboxylase (PDC) with the aim of assessing the contribution of the targeted enzymes to cytoplasmic pH (pH(cyt)) regulation. METHODS: The pH(cyt) was measured by in vivo (31)P nuclear magnetic resonance (NMR) spectroscopy using methyl phosphonate (MeP) as a pH probe. The potential toxicity of MeP was investigated by analysing its effect on the metabolism of radiolabelled glucose. KEY RESULTS: MeP accumulated to detectable levels in the cytoplasm and vacuole of plant tissues exposed to millimolar concentrations of MeP, and the pH-dependent (31)P NMR signals provided a convenient method for measuring pH(cyt) values in tissues with poorly defined signals from the cytoplasmic inorganic phosphate pool. Pretreatment of potato (Solanum tuberosum) tuber slices with 5 mm MeP for 24 h did not affect the metabolism of [U-(14)C]glucose or the pattern of (14)CO(2) release from specifically labelled [(14)C]-substrates. Time-courses of pH(cyt) measured before, during and after an anoxic episode in potato tuber tissues with reduced activities of LDH, or in tobacco (Nicotiana tabacum) leaves with increased activities of PDC, were indistinguishable from their respective controls. CONCLUSIONS: MeP can be used as a low toxicity (31)P NMR probe for measuring intracellular pH values in plant tissues with altered levels of fermentation enzymes. The measurements on transgenic tobacco leaves suggest that the changes in pH(cyt) during an anoxic episode are not dominated by fermentation processes; while the pH changes in the potato tuber tissue with reduced LDH activity show that the affected isozymes do not influence the anoxic pH response.

The effect of molluscan glue proteins on gel mechanics.

Several molluscs have been shown to alternate between a non-adhesive trail mucus and a similar gel that forms a strong glue. The major structural difference between the two secretions is the presence of specific proteins in the adhesive mucus. The present study identifies similar proteins from the glue of the slug Arion subfuscus and the land snail Helix aspersa. To investigate the role played by these proteins in adhesion, the proteins were isolated from the adhesive mucus of different molluscs and added to commercial polymer solutions. The effect was observed qualitatively, and quantified using a dynamic rheometer. The isolated proteins triggered gelling or visible stiffening of agar, pectin and polygalacturonic acid. The effect was stronger on more negatively charged polymers. The effect of the proteins was concentration dependent with an optimal concentration of 1-1.5 mg ml(-1), and was weakened when their structure changed. Other proteins and carbohydrates found in the adhesive mucus had no clear mechanical effect on gels. These findings show that the addition of these proteins to large, anionic polymers plays a central role in the formation of a glue from a mucus-like secretion. Such a mechanism may be common among invertebrates, and it may guide biomimetic approaches in the development of glues and gels.

Effects of Apis mellifera propolis on the activities of streptococcal glucosyltransferases in solution and adsorbed onto saliva-coated hydroxyapatite.

Propolis, a resinous hive product collected by Apis mellifera bees, has been used for thousands of years in folk medicine. Ethanolic extracts of propolis (EEP) have been shown to inhibit the activity of a mixture of crude glucosyltransferase (Gtf) enzymes in solution. These enzymes synthesize glucans from sucrose, which are important for the formation of pathogenic dental plaque. In the present study, the effects of propolis from two different regions of Brazil on the activity of separate, purified Gtf enzymes in solution and on the surface of saliva-coated hydroxyapatite (sHA) beads were evaluated. The EEP from Minas Gerais (MG; Southeastern Brazil) and Rio Grande do Sul (RS; Southern Brazil) were tested for their ability to inhibit the enzymes GtfB (synthesis of insoluble glucan), GtfC (insoluble/soluble glucan) and GtfD (soluble glucan). The effects of propolis on Gtf from Streptococcus sanguis (soluble glucan synthesis) was also explored. The EEP from both regions effectively inhibited the activity of all Gtfs in solution (75-95%) and on the surface of sHA beads (45-95%) at concentrations between 0.75 and 3.0 mg of propolis/ml. However, the two samples of propolis showed different levels of inhibition on each of the enzymes tested. In general, EEP RS demonstrated a significantly higher inhibitory activity on GtfB and C activities (both solution and surface assays) than EEP MG at concentrations between 0.047 and 0.187 mg/ml (p<0.05). EEP MG, on the other hand, exhibited a greater inhibitory effect on the activities of surface GtfD (at 0.375, 0.75 and 1.5 mg/ml) and S. sanguis Gtf (at 1.5 and 3.0 mg/ml; p<0.05). These data indicate that EEP is a potent inhibitor of Gtf enzymes in solution and adsorbed on an experimental pellicle; however, its effect on Gtf activity is variable depending on the geographical origin of the propolis samples. There is a need to identify the active compounds of propolis.

Generational differences in selenium status of women.

In this cross-sectional study of three generations of women, daughters (19-26 yr), mothers (40-58 yr) and maternal grandmothers (67-84 yr) from the same 10 families in central Ohio were studied to determine the effect of life-cycle differences, including estrogen status, on selenium status. Plasma and red blood cell (RBC) selenium and glutathione peroxidase (GPx) activities were determined and typical dietary selenium intakes were calculated from food-frequency questionnaires. Selenium status was lowest in the oldest generation. Plasma selenium of daughters and grandmothers were significantly lower than those of mothers, and plasma GPx and RBC selenium of grandmothers were also lower than those of the mothers. A positive correlation (r = 0.42, p < 0.04) was found between plasma estrogen and plasma selenium concentrations. Selenium intakes of all groups were adequate and no differences in selenium intakes were found among groups. The results of this study indicate that selenium status fluctuates during the female life cycle and is related to estrogen status.

Selenate-supplemented nutritional formula increases plasma selenium in hemodialysis patients.

OBJECTIVE: The purpose of this study was to determine the short-term effect of feeding selenium-supplemented formulas on the selenium status of end-stage renal disease patients on hemodialysis. DESIGN AND SETTING: The prospective, randomized, single-blind study of parallel design was conducted at three hemodialysis clinics. PATIENTS: A total of 79 hemodialysis patients were randomly assigned into one of three groups. INTERVENTION: Liquid nutritional formula supplemented with either selenite (28 microg Se/8 oz, n = 26), selenate (28 microg Se/8 oz, n = 26), or nonfortified (7 microg Se/8 oz, n = 27) was fed to hemodialysis patients as their sole source of nutrition for 14 days. MAIN OUTCOME MEASURE: Plasma and red blood cell (RBC) selenium and glutathione peroxidase (GPX) activities were measured in predialysis blood both before (day 1) and after (day 8) a 7-day baseline period, and after subjects received the formula as the sole source of nutrition (approximately 35 kcal/kg/d) for 14 days (day 22). RESULTS: Selenium intake (Mean +/- SEM, microg/d) was 134 +/- 9, 140 +/- 9, and 35 +/- 2 for patients receiving selenite-, selenate-, or non-supplemented formula, respectively. On day 22, plasma selenium (micromol/L) was greater (P <.032) in the selenate-supplemented group (1.5 +/- 0.1) compared with the nonsupplemented group (1.2 +/- 0.1), but not compared with the selenite-supplemented group (1.4 +/- 0.1). Plasma GPX activity was 44% to 60% that of healthy controls and not different among groups. RBC selenium and GPX activities were within the normal range and were not different among groups. CONCLUSION: The results of this study indicate that a liquid formula supplemented with selenium as selenate is successful at maintaining selenium concentrations within normal range, as well as significantly increasing plasma selenium levels compared with nonsupplementation.

Selective drug delivery to the colon using pectin:chitosan:hydroxypropyl methylcellulose film coated tablets.

A study has been carried out to assess the potential of pectin:chitosan:hydroxypropyl methylcellulose (HPMC) (P:C:H) films for colonic drug delivery. Radiolabelled (99mTc) tablets were coated with a 3:1:1, P:C:H film and administered to human volunteers. The gastro-intestinal transit of the tablets was assessed by gamma scintigraphy. The results showed that in all cases (n=4), the tablets were able to pass through the stomach and small intestine intact. Break up of the tablets commenced once they were in the colon, due to degradation of the coat by colonic bacteria. The study has highlighted the potential of this coating system for colonic drug delivery.

The diverse role of selenium within selenoproteins: a review.

Selenium functions within mammalian systems primarily in the form of selenoproteins. Selenoproteins contain selenium as selenocysteine and perform a variety of physiological roles. Eleven selenoproteins have been identified: cellular or classical glutathione peroxidase; plasma (or extracellular) glutathione peroxidase; phospholipid hydroperoxide glutathione peroxidase; gastrointestinal glutathione peroxidase; selenoprotein P; types 1, 2, and 3 iodothyronine deiodinase; selenoprotein W; thioredoxin reductase; and selenophosphate synthetase. Of these, cellular and plasma glutathione peroxidase are the functional parameters used for the assessment of selenium status. Glutathione peroxidases catalyze the reduction of peroxides that can cause cellular damage. Thioredoxin reductase provides reducing power for several biochemical processes and defends against oxidative stress. Selenoprotein P appears to play a role in oxidant defense. Selenoprotein W may play a role in oxidant defense and be involved with muscle metabolism. Thyroid deiodinases function in the formation and regulation of active thyroid hormone. Selenophosphate synthetase is an enzyme required for the incorporation of selenocysteine into selenoproteins. In addition, a protein in the sperm mitochondrial capsule, which is vital to the integrity of sperm flagella, may be a unique selenoprotein. Recommended intakes, food sources, and status assessment of selenium, as well as selenium's role in health and disease processes, are reviewed.

Cloning, expression, and properties of a nonneuronal secreted acetylcholinesterase from the parasitic nematode Nippostrongylus brasiliensis.

We have isolated a full-length cDNA encoding an acetylcholinesterase secreted by the nematode parasite Nippostrongylus brasiliensis. The predicted protein is truncated in comparison with acetylcholinesterases from other organisms such that the carboxyl terminus aligns closely to the end of the catalytic domain of the vertebrate enzymes. The residues in the catalytic triad are conserved, as are the six cysteines which form the three intramolecular disulfide bonds. Three of the fourteen aromatic residues which line the active site gorge in the Torpedo enzyme are substituted by nonaromatic residues, corresponding to Tyr-70 (Thr), Trp-279 (Asn), and Phe-288 (Met). High level expression was obtained via secretion from Pichia pastoris. The purified enzyme behaved as a monomeric hydrophilic species. Although of invertebrate origin and possessing the above substitutions in the active site gorge residues, the enzyme efficiently hydrolyzed acetylthiocholine and showed minimal activity against butyrylthiocholine. It displayed excess substrate inhibition with acetylthiocholine at concentrations over 2. 5 mM and was highly sensitive to both active site and "peripheral" site inhibitors. Northern blot analysis indicated a progressive increase in mRNA for AChE B in parasites isolated from 6 days postinfection.

Functional magnetic resonance imaging: the basics of blood-oxygen-level dependent (BOLD) imaging.

There are 2 principal techniques of functional MRI (fMRI): the blood-oxygen-level dependent (BOLD) technique, which is the favoured method because no intravenous contrast medium is required, and the dynamic or exogenous technique. The BOLD technique takes advantage of the fact that the change from diamagnetic oxyhemoglobin to paramagnetic deoxyhemoglobin that takes place with brain activation results in decreased signal intensity on MRI. Commercially available scanners can be used to conduct single-slice BOLD fMRI experiments, but echo-planar hardware is needed for multislice wholebrain experiments. Sequence choices in BOLD fMRI include spin-echo and gradient-echo sequences, to which rapid acquisition with relaxation enhancement and echoplanar techniques may be applied. Optimal imaging parameters (echo time, slice thickness, field of view and flip angle) are important in maximizing signal-to-noise ratios. Various statistical techniques and software programs have been developed to interpret the large amounts of data gathered from BOLD fMRI experiments, which presents one of the biggest challenges in performing this technique with clinical MR units. Controversy exists regarding the effects of draining veins on cortical mapping, of inflow of blood into the imaging slice or volume, and of motion artifact. BOLD fMRI has demonstrated good correlation with positron emission tomography, magneto-encephalography and electrocorticographic recordings of motor responses. It has been used to study cortical activity of visual, motor, auditory and speech tasks as well as brain centres for smell, motor imagery, complex motion and memory. As such, it holds promise for the study of brain function, but must be subjected to larger studies comparing it with the gold standard of electrocorticographic mapping.

Binding properties of streptococcal glucosyltransferases for hydroxyapatite, saliva-coated hydroxyapatite, and bacterial surfaces.

The binding specificities of Streptococcus glucosyltransferase (Gtf) B, C and D for hydroxyapatite (HA), saliva-coated hydroxyapatite (SHA), and bacterial surfaces were examined. For HA beads the following values were obtained: (K = affinity; N = number of binding sites) GtfB, K = 46 x 10(5) ml/mumol, N = 0.65 x 10(-6) mumol/m2; GtfC, K = 86 x 10(5) ml/mumol, N = 4.42 x 10(-6) mumol/m2.; GtfD, K = 100 x 10(5) ml/mumol, N = 0.83 x 10(-6) mumol/m2. For SHA beads, the following values were obtained: GtfB, K = 14.7 x 10(5) ml/mumol, N = 1.03 x 10(-6) mumol/m2; GtfC, K = 21.3 x 10(5) ml/mumol, N = 3.66 x 10(-6) mumol/m2; GtfD, K = 1.73 x 10(5) ml/mumol, N = 8.88 x 10(-6) mumol/m2. The binding of GtfB to SHA beads was reduced in the presence of parotid saliva, but the binding of GtfC and D was unaffected. The binding of GtfB to SHA in the presence of parotid saliva supplemented with GtfC and D was reduced when compared with its binding to SHA in the presence of parotid saliva alone. In contrast, te binding of GtfC and SHA was unaffected when parotid saliva was supplemented with the other Gtf enzymes. GtfB bound to several bacterial strains (Strep, mutans GS-5, Actinomyces viscosus OMZ105E and Lactobacillus casei 4646) in an active form, while GtfC and D did not bind to bacterial surfaces. It is concluded that of the three Gtf enzymes, GtfC has the highest affinity for HA and SHA surfaces and can adsorb on the the SHA surface in the presence of the other two enzymes. GtfD also binds to SHA in the presence of the other enzymes but has a very low affinity for the surface. GtfB does not bind to SHA in the presence of the other Gtf enzymes but binds avidly to bacterial surfaces in an active form. Therefore, GtfC most probably binds to apatitic surfaces, while GtfB binds to bacterial surfaces.

Treatment of lymphoid cell malignancy with In-114m labelled autologous lymphocytes.

This paper is a preliminary report of a clinical trial for the treatment of patients with refractory chronic lymphocytic leukaemia, using autologous In-114m-labelled lymphocytes. Fourteen patients have been treated so far with doses ranging from 69 to 211 MBq. All patients had progressive low grade NHL, resistant to chemotherapy and conventional radiotherapy. Following the intravenous administration of radiolabelled autologous lymphocytes 53% (range 33-92%) of the activity accumulated in the spleen, 35% (21-64%) in the liver and 5% in the bone marrow. The initial response in all patients was a rapid decrease in lymphocyte count in peripheral blood. 10 of the 14 (72%) patients showed a response to the treatment. In 2 patients, there was a complete response which lasted 24 and 36 months respectively, 8 patients showed a partial response of 2 to 17 months duration. None of the patients experienced any subjective toxicity although myelosuppression was seen in all patients. This is a novel concept for the administration of therapeutic radiation in a selective way for the treatment of lymphoid cell malignancy and has produced significant antitumour effect in patients with highly resistant disease. The trial is ongoing and a full report will be published on its completion.

Psychological impact of injuries in athletes.

Although research on the psychological impact of injury is in its infancy, this article reviews relevant literature focusing on post-injury emotional response, self-esteem, and the effect of mood disturbance on rehabilitation from sport injury. Injury is often accompanied by depression, tension, anger and low self-esteem, particularly in competitive, seriously injured athletes. Mood disturbance seems to relate to the athlete's perceived progress in rehabilitation and has been shown to negatively relate to attendance at rehabilitation sessions. This article also describes how the Emotional Responses of Athletes to Injury Questionnaire (ERAIQ) serves as a guide for the initial interview of an injured athlete. Interventions such as positive self-talk, relaxation, goal setting and healing imagery, all used by a faster healing group of athletes, and although not well researched, seem appropriate to assist athletes in coping with injury. Modelling interventions during injury rehabilitation have also been shown to have a positive effect on rehabilitation and should be used. These relationships are described in more depth and in the context of a theoretical model. Directions for future research are suggested.