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Effect of eccentric exercise on muscle oxidative metabolism in humans.

PURPOSE: The purpose of this study was to evaluate the effects of eccentric exercise on muscle oxidative function. METHODS: Thirteen subjects performed high-intensity eccentric cycling for 30 min. Muscle oxidative function in vastus lateralis was evaluated by measurements of respiration in permeabilized muscle fibers (skinned fibers) and from the kinetics of oxyhemoglobin (oxyHb) saturation measured with near infrared spectroscopy (NIRS). RESULTS: After eccentric cycling, all subjects reported extensive delayed onset muscle soreness (DOMS), but plasma markers of muscle damage (creatine kinase and beta-glucuronidase activity) were not significantly altered. The half time of oxyHb desaturation after circulatory occlusion (128 +/- 11 s, mean +/- SE) and oxyHb resaturation after restoration of blood flow (13.8 +/- 0.7 s) were not significantly changed after eccentric cycling (N = 7). Respiration in skinned muscle fibers measured in the absence of ADP and in the presence of a submaximal (0.1 mM) or maximal ADP concentration (1 mM) was not significantly changed after eccentric cycling (N = 6). The sensitivity of respiration to ADP was not significantly changed after eccentric cycling. CONCLUSIONS: Muscle oxidative function (maximal respiration and respiratory control by ADP) was not compromised after high-intensity eccentric cycle exercise. Furthermore, NIRS indicates that after eccentric cycling muscle oxygen utilization and local oxygen transport at rest are unchanged. It is concluded that eccentric cycling, although causing DOMS, does not negatively affect skeletal muscle oxidative function.

Adult↗

The transcription factor Sox10 is a key regulator of peripheral glial development.

The molecular mechanisms that determine glial cell fate in the vertebrate nervous system have not been elucidated. Peripheral glial cells differentiate from pluripotent neural crest cells. We show here that the transcription factor Sox10 is a key regulator in differentiation of peripheral glial cells. In mice that carry a spontaneous or a targeted mutation of Sox10, neuronal cells form in dorsal root ganglia, but Schwann cells or satellite cells are not generated. At later developmental stages, this lack of peripheral glial cells results in a severe degeneration of sensory and motor neurons. Moreover, we show that Sox10 controls expression of ErbB3 in neural crest cells. ErbB3 encodes a Neuregulin receptor, and down-regulation of ErbB3 accounts for many changes in development of neural crest cells observed in Sox10 mutant mice. Sox10 also has functions not mediated by ErbB3, for instance in the melanocyte lineage. Phenotypes observed in heterozygous mice that carry a targeted Sox10 null allele reproduce those observed in heterozygous Sox10(Dom) mice. Haploinsufficiency of Sox10 can thus cause pigmentation and megacolon defects, which are also observed in Sox10(Dom)/+ mice and in patients with Waardenburg-Hirschsprung disease caused by heterozygous SOX10 mutations.

Animals↗

Significance of nodal metastatic tumor characteristics in nodal metastasis and prognosis of patients with invasive ductal carcinoma of the breast.

There is no study evaluating the significance of nodal metastatic tumor characteristics in tumor progression of invasive ductal carcinomas (IDCs) of the breast. The purpose of this study was to investigate whether nodal metastatic tumor characteristics play an important role in the tumor progression of IDCs. The subjects of this study were 205 IDC patients with nodal metastases. Significant associations with increased numbers of nodal metastases, and patient outcomes were evaluated by multivariate analyses, in comparison with well-known histological parameters. The numbers of lymph nodes with extra-nodal invasion and with extranodal blood vessel tumor emboli, the distance of extra-nodal blood vessel tumor emboli from the nodes, and the nodal tumor dimensions significantly increased the number of nodal metastases in the multivariate analysis (P<0.001). Cox multivariate analyses showed that the parameters which significantly increased hazard rates (HRs) of disease-free survival (DFS), distant organ metastasis (DOM) and overall survival were 6 or more mitotic figures of nodal metastatic tumors (P<0.05). Six or more lymph nodes with extra-nodal invasion, and an extra-nodal blood vessel tumor emboli dimension of >0.6 mm significantly increased the HRs of DFS and DOM in multivariate analyses (P<0.05). The present study demonstrated the important roles of nodal metastatic tumors in the tumor progression of IDCs.

Adult↗

Energy evaluation of extruded compound foods for dogs by near-infrared spectroscopy.

Near-infrared reflectance spectroscopy (NIRS) was used to predict the chemical composition, apparent digestibility and digestible nutrients and energy content of commercial extruded compound foods for dogs. Fifty-six foods of known chemical composition and in vivo apparent digestibility were analysed overall and 51 foods were used to predict gross energy digestibility and digestible energy content. Modified partial least square calibration models were developed for organic matter (OM), crude protein (CP), ether extract (EE), crude fibre (CF), nitrogen free extracts (NFE) and gross energy (GE) content, the apparent digestibility (OMD, CPD, EED, NFED and GED) and the digestible nutrient and energy content (DOM, DCP, DEE, DNFE and DE) of foods. The calibration equations obtained were evaluated by the standard error and the determination coefficient of cross-validation. The cross-validation coefficients of determination (R) were 0.61, 0.99, 0.91, 0.96, 0.94 and 0.92 for OM, CP, EE, CF, NFE and GE, the corresponding standard error of cross-validation (SECV) being 5.80, 3.51, 13.35, 3.64 and 16.95 g/kg dry matter (DM) and 0.29 MJ/kg DM respectively. The prediction of apparent digestibility was slightly less accurate, but NIRS prediction of digestible nutrient (g/kg DM) and DE (MJ/kg DM) gave satisfactory results, with high R (0.93, 0.97, 0.93, 0.83 and 0.93 for DOM, DCP, DEE, DNFE and DE respectively) and relatively low SECV (11.55, 6.85, 12.14 and 22.98 g/kg DM and 0.47 MJ/kg DM). It is concluded that the precision of NIRS in predicting the energy value of compound extruded foods for dogs is similar or better than by proximate analysis, as well as being faster and more accurate.

Animal Feed↗

Algae-bacteria interactions and their effects on aggregation and organic matter flux in the sea.

Aggregation of algae, mainly of diatoms, is an important process in marine pelagic systems, often terminating phytoplankton blooms and leading to the sinking of particulate organic matter in the form of marine snow. This process has been studied extensively, but the specific role of heterotrophic bacteria has largely been neglected, mainly because field studies and most experimental work were performed under non-axenic conditions. We tested the hypothesis that algae-bacteria interactions are instrumental in aggregate dynamics and organic matter flux. A series of aggregation experiments has been carried out in rolling tanks with two marine diatoms typical of temperate regions (Skeletonema costatum and Thalassiosira rotula) in an axenic treatment and one inoculated with marine bacteria. Exponentially growing S. costatum and T. rotula exhibited distinctly different aggregation behavior. This was reflected by their strikingly different release of dissolved organic matter (DOM), transparent exopolymer particles (TEP) and protein-containing particles (CSP), as well as their bacterial biodegradability and recalcitrance. Cells of S. costatum aggregated only little and their bacterial colonization remained low. Dissolved organic matter, TEP and CSP released by this alga were largely consumed by free-living bacteria. In contrast, T. rotula aggregated rapidly and DOM, TEP and CSP released resisted bacterial consumption. Experiments conducted with T. rotula cultures in the stationary growth phase, however, showed rapid bacterial colonization and decomposition of algal cells. Our study highlights the importance of heterotrophic bacteria to control the development and aggregation of phytoplankton in marine systems.

Bacteria↗

Characterization of the 5-HT receptor subtypes involved in the motor behaviours produced by intrathecal administration of 5-HT agonists in rats.

1. The motor behavioural effects of intrathecal injections of 5-hydroxytryptamine (5-HT) and a variety of 5-HT receptor agonists were examined in adult Wistar rats to establish; (a) which 5-HT receptor subtype/s elicit each behaviour and (b) whether these receptors are located within the spinal cord. 2. Intrathecal injection of 5-methoxy-N,N'-dimethyltryptamine (5-MeODMT), (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride (DOI) or 2,5-dimethoxy-alpha,4-dimethylbenzene ethamine hydrochloride (DOM) produced dose-related back muscle contractions (BMC) and wet dog shakes (WDS) which were both markedly attenuated by intraperitoneal pretreatment with either ritanserin (1 mg kg-1), ketanserin (0.16 mg kg-1) or mianserin (0.6 mg kg-1) indicating the involvement of 5-HT2 receptors in both these motor behaviours. Both fluoxetine (1-20 mg kg-1, i.p.) and high doses of 5-HT (50 micrograms) following fluoxetine (5 mg kg-1, i.p.) also elicited BMC, further confirming the involvement of 5-HT in this behaviour. 3. Intrathecal 5-carboxamidotryptamine (5-CT) evoked a marked wet-dog shake response without producing any BMC. Intrathecal pretreatment with 8-hydroxy-2-(di-n-propylamino)tetraline (8-OH-DPAT) enhanced, while in contrast 2-methyl-5-HT pretreatment attenuated, 5-HT agonist-induced BMC without affecting WDS. These data suggest that the spinal 5-HT2 receptors mediating BMC are positively modulated by 5-HT1A but negatively influenced by 5-HT3 receptor activation and may be of a different subtype to the supra-spinal 5-HT2 receptors which elicit WDS. 4. A contrast, reciprocal forepaw treading, lateral head weaving, flat body posture and Straub-tail were evoked by 5-MeODMT, 8-OH-DPAT or 5-CT but not by DOI or DOM indicating that these behaviours were not produced by 5-HT2 receptor activation alone. Ritanserin (1 mg kg- 1, i.p.) or ketanserin (0.16mgkg-1, i.p.) pretreatment reduced the reciprocal forepaw treading induced by high intrathecal doses of either 5-MeODMT (25.pg) or 5-CT (50,ug) suggesting that this behaviour may be facilitated by 5-HT2 receptor activation. 5. Intrathecal injection of 5-HT (0.05-50pg, after systemic fluoxetine, 5mg kg 1, i.p.), or 1-(3-chlorophenyl) piperazine (mCPP) produced dose-related forepaw-licking and grooming, neither of which were attenuated by ketanserin (0.16 mgkg-1, i.p.) pretreatment suggesting these behaviours may be mediated by 5-HT1c receptors. In contrast, 2-methyl-5-HT (50 and 100pg) produced sideward tail-flicks, not evoked by any other 5-HT agonist and could therefore be mediated by spinal 5-HT3 receptor activation. 6. These data provide behavioural evidence for the existence of spinal 5-HT2 receptors which produce a novel motor behaviour, BMC. Ligand binding studies and dose-response studies with a range of selective 5-HT antagonists are required to establish whether BMC and WDS are mediated by different subtypes of 5-HT2 receptors.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

Characterization of the discriminative stimulus properties induced by 5-HT1 and 5-HT2 agonists in rats.

The effect of different serotonin (5-HT) agonists and antagonists on the discriminative stimulus properties (cue) induced by 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OHDPAT), 1-(m-trifluoromethylphenyl)piperazine (TFMPP) and d-LSD (d-lysergic acid diethylamide) has been investigated. The 8-OHDPAT cue was mimicked by the 5-HT1A agonists ipsapirone, buspirone, gepirone and partially by 5-methoxy-N,N-dimethyltryptamine and d-LSD. 5-HT1B (TFMPP and RU 24969) and 5-HT2 agonists (DOM, DOI and quipazine) were ineffective and induced disruption of responding. The 8-OHDPAT cue was antagonized by spiroxatrine and partially by (-)-alprenolol, whereas selective antagonists of 5-HT2 (ketanserin and ritanserin), 5-HT3 (ICS 205-930), alpha 1-adrenergic (prazosin) and beta-adrenergic receptors (ICI 118.551) were ineffective. The TFMPP cue was mimicked by RU 24969 and partially by quipazine. Other compounds were ineffective. Only (-)-alprenolol antagonized the effect of TFMPP. The d-LSD cue was mimicked by DOM, DOI, quipazine, 5-methoxy-N,N-dimethyltryptamine and partially by ipsapirone, TFMPP and RU 24969. The 3 latter compounds and 5-HT1A agonists induced disruption of responding. The d-LSD cue was antagonized by ketanserin and ritanserin, but not by the other antagonists mentioned above. The specific inhibitor of 5-HT uptake citalopram was not able to substitute for any of the 3 agonists. It is concluded that the drug discrimination technique can be used to identify selective agonists and antagonists of 5-HT receptor subtypes. Compounds with mixed effects on 5-HT receptor subtypes can also be identified. These show additional effects on reaction time and often disrupt responding at higher dosages.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

The effects of preexercise stretching on muscular soreness, tenderness and force loss following heavy eccentric exercise.

The present study sought to investigate the effects of preexercise stretching on delayed onset muscle soreness (DOMS), i.e. soreness, tenderness and loss of muscle force, that usually occurs after strenuous or unaccustomed eccentric exercise. Ten female volunteers performed 10 sets of 10 maximal isokinetic eccentric contractions for knee flexion with both legs after a 5-min ergometer cycling warm-up. Prior to the exercise for one leg, randomly chosen, 4 x 20 s of static stretching for the hamstring muscle group was implemented. Rated soreness, tenderness on algometer pressure and loss of maximal eccentric contractile force was evaluated preexercise and 24, 48 and 96 h postexercise. The exercise bout produced severe DOMS, with parameters peaking and troughing at 48 h postexercise. However, no significant differences were found, regarding any of the parameters, when comparing stretched and nonstretched legs. The present study thus suggests that preexercise static stretching has no preventive effect on the muscular soreness, tenderness and force loss that follows heavy eccentric exercise.

Adult↗

Muscular mechanical hyperalgesia revealed by behavioural pain test and c-Fos expression in the spinal dorsal horn after eccentric contraction in rats.

Delayed onset muscle soreness (DOMS) is quite common, but the mechanism for this phenomenon is still not understood; even the existence of muscle tenderness (mechanical hyperalgesia) has not been demonstrated in experimental models. We developed an animal model of DOMS by inducing eccentric contraction (lengthening contraction, ECC) to the extensor digitorum longus muscle (EDL), and investigated the existence of mechanical hyperalgesia in the EDL by means of behavioural pain tests (Randall-Selitto test and von Frey hair test, applied to/through the skin on the EDL muscle) and c-Fos expression in the spinal dorsal horn. We found that the mechanical withdrawal threshold measured with the Randall-Selitto apparatus decreased significantly between 1 and 3 days after ECC, while that measured by von Frey hairs did not. The group that underwent stretching of the muscle only (SHAM group) showed no change in mechanical pain threshold in either test. These results demonstrated that the pain threshold of deep tissues (possibly of the muscle) decreased after ECC. c-Fos immunoreactivity in the dorsal horn (examined 2 days after ECC/SHAM exercise) was not changed by either ECC or compression (1568 mN) to the EDL muscle by itself, but it was significantly increased by applying compression to the EDL muscle 2 days after ECC. This increase was observed in the superficial dorsal horn of the L4 segment of the ipsilateral side, and was clearly suppressed by morphine treatment (10 mg kg(-1), i.p.). These results demonstrated the existence of mechanical hyperalgesia in the muscle subjected to ECC. This model could be used for future study of the neural mechanism of muscle soreness.

Animals↗

Stem cell self-renewal controlled by chromatin remodeling factors.

The self-renewing ability of a stem cell is controlled by its specialized micro-environment or niche, whereas epigenetic regulation of gene expression by chromatin remodeling factors underlies cell fate determination. Here we report that the adenosine triphosphate-dependent chromatin remodeling factors ISWI and DOM control germline stem cell and somatic stem cell self-renewal in the Drosophila ovary, respectively. The iswi mutant germline stem cells are lost rapidly because of defects in responding to bone morphogenetic protein niche signals and in repressing differentiation, whereas the dom mutant somatic stem cells are lost because of defective self-renewal. This work demonstrates that different stem cell types can use different chromatin remodeling factors to control cell self-renewal.

Adenosine Triphosphatases↗

Rapid Recovery of Marine Bacterioplankton Activity after Inhibition by UV Radiation in Coastal Waters.

Laboratory and in situ experiments were performed in order to evaluate the role of UV radiation on bacterial activity. Particular attention was given to the determination of the role of UV-A and photosynthetic active radiation (PAR) and different nutrient conditions on the recovery of bacterial activity. Laboratory experiments with nearly natural radiation intensities indicated a 20 to 40% reduction from the initial level of bacterial activity after UV-B exposure for 2 to 4 h. Bacterial activity in freshly collected seawater showed a more pronounced inhibition and faster recovery than bacterial activity in aged, nutrient-depleted seawater. The results of in situ experiments with filtered water (0.8-(mu)m-pore-size filter) and natural surface solar radiation levels agreed with those of the laboratory experiments and revealed that UV-A and PAR are important for the recovery of bacterial activity and result in levels of bacterial activity that are higher than those prior to exposure to full solar radiation. Bacterioplankton exposed to full solar radiation for 3 h and subsequently incubated at different depths within the upper mixed water column showed an increase in bacterial activity with increased depth; the highest bacterial activity was detected at depths of 5.5 to 10.5 m, where the short-wavelength UV-B was already largely attenuated, but enough long wavelength UV-A and short PAR were available to allow recovery. This elevated bacterial activity following exposure to UV-B was attributed to the photolysis of dissolved organic matter (DOM) exposed to near-surface radiation and to the rapid recovery of bacteria from UV stress once they were mixed into deeper layers of the upper mixed water column, where they efficiently utilize the photolytically cleaved DOM. It is concluded that studies on the role of UV on the carbon and energy flux through the upper layer of the ocean should take into account the highly dynamic radiation conditions.

Journal Article↗

Isolation and characterization of mutant strains of Bordetella bronchiseptica lacking dermonecrotic toxin-producing ability.

Mutant strains of Bordetella bronchiseptica, named B-42, B-76, B-84, and B-119, were obtained after serial passages of a parent strain, L3, on Bordet-Gengou agar plates containing 20% horse blood and 200 micrograms of nalidixic acid per ml (BGN-20 agar plates) at 42 degrees C. Mutant strains completely lacked dermonecrotic toxin-producing ability, and lethal activity of the strains for mice was apparently reduced compared with that of strain L3. Mutant strains were able to grow at 42 degrees C, and the strains were nalidixic acid resistant. The mutant strains showed domed (Dom+) colony morphology with smooth texture (Scs+) and no production of zone of hemolysis (Hly-), but the agglutinability of these strains to antiserum prepared with Dom+ Scs+ Hly+ organisms of strain L3 was the same as that of strain L3. When strain B-42 was inoculated intramuscularly or intranasally into guinea pigs, all the animals survived without manifesting clinical signs and produced a high-level of serum agglutination antibodies against strain L3. These inoculated animals were protected against intranasal challenge with strain L3. These properties of mutant strains are hereditarily stable after 50 subcultures on BGN-20 agar plates or 20 passages in mice. These data suggest that the mutant strains lacking dermonecrotic toxin-producing ability can be used as a live attenuated vaccine against swine atrophic rhinitis.

Animals↗

The effects of massage on delayed onset muscle soreness.

OBJECTIVES: The purpose of this study was to investigate the physiological and psychological effects of massage on delayed onset muscle soreness (DOMS). METHODS: Eighteen volunteers were randomly assigned to either a massage or control group. DOMS was induced with six sets of eight maximal eccentric contractions of the right hamstring, which were followed 2 h later by 20 min of massage or sham massage (control). Peak torque and mood were assessed at 2, 6, 24, and 48 h postexercise. Range of motion (ROM) and intensity and unpleasantness of soreness were assessed at 6, 24, and 48 h postexercise. Neutrophil count was assessed at 6 and 24 h postexercise. RESULTS: A two factor ANOVA (treatment v time) with repeated measures on the second factor showed no significant treatment differences for peak torque, ROM, neutrophils, unpleasantness of soreness, and mood (p > 0.05). The intensity of soreness, however, was significantly lower in the massage group relative to the control group at 48 h postexercise (p < 0.05). CONCLUSIONS: Massage administered 2 h after exercise induced muscle injury did not improve hamstring function but did reduce the intensity of soreness 48 h after muscle insult.

Adult↗

Evaluation of six oxygen concentrators.

Examples of six oxygen concentrators (DeVO2, Dom 10, Econo 2, Hudson, Permox, and Roomate) were evaluated over a 9-28 day period to determine (1) the oxygen yield (% O2) over the flow range 1-4 l min-1; (2) 90% oxygen rise time (90% RT) from a cold start when they were operated at 2 l min-1; (3) accuracy and readability of the flow device; (4) static outlet pressure; (5) major components comprising the product gas (Hudson only); and (6) general characteristics. At an outlet flow of 2 l min-1 the mean % O2 generated by all models, except the Permox (which was lower, mean (SD) 90.5% (3.1%), were between 94% and 95% with a range of less than +/- 0.5%. The Dom 10, Econo 2, and Hudson consistently generated higher oxygen concentrations than the other models at flow rates greater than 2 l min-1. The 90% RT was less than 10.5 minutes for all models. Deviations of up to 22% were observed between predicted and measured flow rates in all models except the DeVO2, Hudson, and Permox. It was possible to set the orifice type flow devices fitted to the Permox and Roomate between indicated flow settings, resulting in cessation of flow. Spectral analysis of the output of one device showed that argon and oxygen were concentrated to similar extents, indicating that the maximal attainable oxygen yield for a molecular sieve concentrator is about 96%.

Evaluation Studies as Topic↗

Light concentric exercise has a temporarily analgesic effect on delayed-onset muscle soreness, but no effect on recovery from eccentric exercise.

This study investigated the hypothesis that a bout of light concentric exercise (LCE) would alleviate delayed-onset muscle soreness (DOMS) and enhance recovery from muscle damage. Fourteen subjects performed two bouts of 60 maximal eccentric actions of the elbow flexors (Max-ECC) separated by 2-4 weeks. One arm performed LCE (600 elbow flexion and extension actions with minimal force generation) 1, 2, 3, and 4 d after Max-ECC; the contralateral (control) arm performed only Max-ECC. Changes in maximal isometric and isokinetic strength, range of motion (ROM), upper arm circumference, and muscle soreness and tenderness were assessed before and immediately after LCE bouts. Changes in these measures and plasma creatine kinase (CK) activity for 7 d after Max-ECC were compared between the control and LCE arms using 2-way repeated measures analysis of variance (ANOVA). Significant (p < 0.05) decreases in muscle soreness (approximately 40%) and tenderness (approximately 40%) were evident immediately after LCE, which also resulted in small but significant decreases in strength (approximately 15%) and increases in ROM (approximately 5 degrees ). No significant differences in the changes in the measures following Max-ECC were observed between the arms. These results suggest that LCE has a temporary analgesic effect on DOMS, but no effect on recovery from muscle damage.

Adaptation, Physiological↗

Comparison between old and young men for changes in makers of muscle damage following voluntary eccentric exercise of the elbow flexors.

This study aimed to investigate if old men were more susceptible than young men to muscle damage induced by exercise consisting of repeated-lengthening muscle actions. The responses to a bout of eccentric exercise were compared between 10 young (mean age +/- SEM = 19.4 +/- 0.4 y) and 10 old (70.5 +/- 1.5 y) men. All subjects performed 6 sets of 5 lengthening actions of the left elbow flexors at a range of 90 degrees from an elbow flexed (90 degrees ) to an extended (180 degrees ) position in 5 s using a dumbbell massed at 40% maximal isometric strength (MVC) at an elbow joint angle of 90 degrees . Changes in MVC, range of motion (ROM), upper arm circumference (CIR), muscle soreness (DOMS), plasma creatine kinase activity (CK), and myoglobin (Mb) concentration over 7-10 d following exercise were compared between groups by 2-way repeated measures analysis of variance (ANOVA). Significant differences between groups were evident at baseline for ROM (significantly smaller for the older group) and CIR (significantly larger for the older group), but not for MVC and other measures. Contrary to the hypothesis, the young group showed significantly larger decreases in MVC and ROM and larger increases in circumference, DOMS, CK activity, and Mb con centration than those of the old group. These results suggest that muscle damage is not necessarily greater in old versus young men following voluntary eccentric exercise. It may be that physiological changes that occur with ageing, including a decrease in ROM, reduce damaging stress to muscles during lengthening muscle actions.

Adult↗

Dissolved organic matter utilization and oxygen uptake in algal-bacterial microcosms.

Under closed laboratory conditions, at non-limiting nutrient levels, the biomass of Anabaena variabilis, Anacystis nidulans, Chlorella pyrenoidosa, and Selanastrum capricornutum increased with increasing levels of dissolved organic matter (DOM) as a result of bacterially produced carbon dioxide (CO2) and (or) cofactors. Oxygen (O2) produced as a result of algal photosynthesis was sufficient to supply the najority of O2 required by the bacterial community. The percentage of DOM utilized by bacteria which was subsequently incorporated into algal biomass varied with individual species indicating that the association between individual algal species and the bacterial microbiota varied. Under natural conditions bacteria could provide CO2 and (or) cofactors for algal photosynthesis which in turn supplies O2 for bacterial respiration. This mutualistic association in aquatic environments could result in an increase in planktonic and epiphytic algal biomass if other nutrients are available.

Bacteria↗

DNA fusion vaccines induce epitope-specific cytotoxic CD8(+) T cells against human leukemia-associated minor histocompatibility antigens.

The graft-versus-leukemia effect of allogeneic stem-cell transplantation is believed to be mediated by T-cell recognition of minor histocompatibility antigens on recipient cells. For minor histocompatibility antigens HA-1 and HA-2, normal cell expression is restricted to hemopoietic cells, and boosting the immune response to these antigens may potentiate graft-versus-leukemia effect without accompanying graft-versus-host disease. To increase efficacy, expansion of HA-1- or HA-2-specific CTL before transplantation is desirable. However, primary HA-1- or HA-2-specific CTL expanded in vitro are often of low avidity. An alternative approach is to prime specific CTL responses in vivo by vaccination. Clearly, donor vaccination must be safe and specific. We have developed DNA fusion vaccines able to induce high levels of epitope-specific CTL using linked CD4(+) T-cell help. The vaccines incorporate a domain of tetanus toxin (DOM) fused to a sequence encoding a candidate MHC class I binding peptide. This design generates antitumor CD8(+) T-cell responses and protective immunity in preclinical models. For clinical application, we constructed vaccines encoding HLA-A*0201-restricted peptides from human HA-1 and HA-2, which were fused to DOM, and tested their performance in HLA-A*0201-transgenic mice. Priming induced epitope-specific, IFNgamma-producing CD8(+) T cells with cytotoxic function boosted to high levels with electroporation. Strikingly, these mouse T cells efficiently killed human lymphoblastoid cell lines expressing endogenous HA-1 or HA-2. High avidity is indicated by the independence of cytolysis from CD8/MHC class I interaction. These safe epitope-specific vaccines offer a potential strategy to prime HA-1- or HA-2-specific CTL in transplant donors before adoptive transfer.

Amino Acid Sequence↗