Oxidative stress biomarkers response to high intensity interval training and relation to performance in competitive swimmers.

AIM: Aim of the study is to investigate the modulations of oxidative stress biomarkers and some antioxidants induced by high intensity interval training bout and its relation to swimming performance. METHODS: Ten swimmers performed a set of 8 maximal swims along 100 m by style of their specialty, with 10 minute for a rest. The concentration of blood lactate ([Lac]) was determined after each swim. The lactate tolerance index (LTI) was determined by the ratio between [Lac] and the respective times of execution of the 8 swims. The time to complete first 100 m swim at maximum effort (P100) and the international point score (IPS) reached in a specific competition were considered performance parameters. Venous blood was collected before and after the anaerobic training effort. RESULTS: Mean blood lactate concentration in the eight swims was 10.9 ± 1.2 mM. Significant increases were observed for TBARS (pre: 4.1±0.7 ?mol/L; post: 4.9±1.1. ?mol/L), CK (pre: 206.4±170.7 U/L; post: 244.4±176.9. U/L), GSH (pre: 0.52±0.06; post: 0.62±0.05. mM), and ascorbic acid (pre: 0.06±0.02; post: 0.11±0.03. mg/dL) after the anaerobic training bout compared to the values obtained before it. In addition, significant correlations (P < 0.05) were detected between LTI and P100 (r = -0.87) and IPS (r = 0.64) and between variation of ascorbic acid and P100 (r = -0.60). CONCLUSION: Anaerobic training bout proposed induces oxidative stress and cell muscle damage markers as well as modulates some antioxidants of competitive swimmers. The modulation of ascorbic acid seems to play an important role in the performance of these athletes.

Thrombin reduces cerebral arterial contractions caused by cerebrospinal fluid from patients with subarachnoid hemorrhage.

BACKGROUND AND PURPOSE: We observed that the second application of cerebrospinal fluid (CSF) from subarachnoid hemorrhage (SAH) patients onto cerebral arterial segments in vitro produces a greater contraction than does the initial application. It was hypothesized that the difference between the first and second applications of SAH CSF was due to the activity of thrombin. METHODS: Canine vertebrobasilar artery was removed under general anesthesia, cut into rings, and suspended in tissue culture baths so as to measure isometric tension. CSF was taken from patients 1 to 3 days after SAH via ventricular drains. CSF was administered in 10(-5) to 10(-1) dilutions. The thrombin antagonist hirudin (5 U) was administered before CSF in some experiments. The arterial tension response to pure oxyhemoglobin (10(-4) to 3.2 g/dL) and thrombin (10(-4) to 3.2 U/mL), administered alone or in combination, was also examined. RESULTS: Hirudin increased arterial tension generated on the initial application of SAH CSF but had no effect on the tension generated by the second application of the SAH CSF, suggesting that thrombin limits the tension generated by vasoconstrictive agents in the CSF. Thrombin and pure oxyhemoglobin administered together produced less tension than that generated in response to oxyhemoglobin administered alone; no additive response was observed by coadministering the 2 vasoconstrictive agents. CONCLUSIONS: In the presence of oxyhemoglobin, thrombin acts to reduce cerebral arterial tension. This interaction between thrombin and hemoglobin may account for the observation that the second application of CSF from SAH patients onto cerebral arterial segments in vitro produces a greater contraction than does the initial application.

A polysomnographic study of sleep disturbance in community elderly with self-reported environmental chemical odor intolerance.

Subjective sleep complaints and food intolerances, especially to milk products, are frequent symptoms of individuals who also report intolerance for low-level odors of various environmental chemicals. The purpose of the present study was to evaluate the objective nature of nocturnal sleep patterns during different diets, using polysomnography in community older adults with self-reported illness from chemical odors. Those high in chemical odor intolerance (n = 15) exhibited significantly lower sleep efficiency (p = .005) and lower rapid-eye-movement (REM) sleep percent (p = .04), with a trend toward longer latency to REM sleep (p = .07), than did those low in chemical intolerance (n = 15), especially on dairy-containing as compared with nondairy (soy) diets. The arousal pattern of the chemical odor intolerant group differed from the polysomnographic features of major depression, classical organophosphate toxicity, and subjective insomnia without objective findings. The findings suggest that community elderly with moderate chemical odor intolerance and minimal sleep complaints exhibit objectively poorer sleep than do their normal peers. Individual differences in underlying brain function may help generate these observations. The data support the need for similar studies in clinical populations with chemical odor intolerance, such as multiple chemical sensitivity patients and perhaps certain veterans with "Persian Gulf Syndrome."

Suppression of CTL responses in vitro by large granular T cells.

The generation of large granular T cells (LGC) that display cytotoxicity similar to activated natural killer (NK) cells was described previously by the authors. To determine whether LGC can regulate the generation of allospecific CTL (C57BL/6, anti-DBA/2), we used primed responding T cells (Thy-1.2) and (Thy-1.1) to perform co-cultures. The cytolytic activities and the phenotype of the harvested T cells were examined after a co-culture period (3-6 days). We observed that the survival of fresh responding T cells was impaired in the co-cultures. Activated (4 days) responding T cells can survive in similar coculture conditions. This led to the conclusion that LGC can inactivate the CTL generation in fresh T-cell populations and that fresh and activated responding cells have different susceptibilities to LGC interactions.

A peptide derived from melanocytic protein gp100 and presented by HLA-B35 is recognized by autologous cytolytic T lymphocytes on melanoma cells.

A panel of autologous cytolytic T lymphocyte (CTL) clones have been isolated from blood lymphocytes of a melanoma patient after in vitro stimulation with autologous tumor cells. We previously reported the molecular definition of three distinct antigens recognized by some of these CTL clones. We describe here, the identification of a fourth antigenic peptide expressed by this melanoma line and recognized by a CTL clone restricted by HLA-B*3503. The antigenic peptide, which is nine-amino acid long, has the sequence LPHSSSHWL and is derived from melanocyte differentiation antigen gp100. As HLA-B35 is one of the most frequent HLA-B alleles, being present in 20% of the Caucasian individuals, this peptide may be a good target for peptide-based immunotherapy of melanoma.

[In vitro comparison of 2 expressions of cellular immunity induced by a cutaneous allograft in the mouse]. / Comparaison in vitro de deux expressions de l'immunité cellulaire induite par une allogreffe cutanée chez la souris

A short-term test (5 h) has been compared to a long-term test (48 h) for evaluation of the cellular cytotoxic response which is induced in recipients of allografts. Spleen cells from mice recipients of skin allografts were tested. We have shown that the cellular cytotoxicity expressed in a short-term test is itself expressed during a short period whereas the cytotoxicity expressed in a long-term test is still detectable for months after a first skin graft. Moreover, the lytic activity of the spleen cells is much higher when it is expressed in a long-term test. The fact that we could get, in vitro, in two days, a low primary cellular cytotoxic response against alloantigens, makes likely that during the long-term test, the anamnestic response which can develop is especially responsible for the cellular cytotoxic response.

Suppression of cytolytic T-lymphocyte responses through inactivation of non-T accessory cells.

The suppressive properties of nonspecific cytotoxic T lymphocyte (CTL) populations, derived from murine fetal calf serum (FCS)-precultured cells expanded in interleukin 2 (operationally defined as FCS-CM-expanded cells), were investigated on CTL responses generated by syngeneic alloreactive lymphoid cells. Our results suggest that the addition of FCS-CM-expanded cell populations can inhibit the CTL response by elimination of the bone marrow-derived macrophage (BM M phi) population used as non-T accessory cells. Indeed, in the culture conditions used, removal of IL-2 by the FCS-CM-expanded cells as well as a direct inactivating effect on the CTL precursors (CTL-P) could be excluded as a reason for inhibition. On the other hand, we were able to show that the BM M phi population was very sensitive to the cytolytic activity exhibited by the inhibiting cells in a 3 h 51Cr-release assay and that the suppressor effect observed could be partially circumvented by a second addition of BM M phi on the second day after the initiation of the culture.

Ultrastructural study of the relationship between immune cytotoxic lymphoid cells and target cells in vitro.

Ultrastructural details of the relationship in vitro between sensitized T lymphoid cells and YAC target cells are described. Within a few hours very close connections develop between these two cells. Narrow junctions with parallel plasma membranes and local evaginations of the lymphoid cell into recesses of the target cell are specific aspects. Very fine threadlike material is found between the two plasma membranes at the narrowest zones. The lymphoid cells concerned have an intermediate structure between ordinary small lymphocytes and blast cells. They do not seem to be modified in the process while YAC target cells are severly damaged. Polysomes are dispersed; mitochondria and rough endoplasmic reticulum are affected locally or generally. Later, nucleoli and chromatin become dense and pycnosis occurs. The significance of these results is discussed.

FCS-activated blast T cells inhibit the in vitro response of memory CTL precursors to alloantigens by removal of factor(s) from MLC supernatant.

The inhibitory effect of spleen cells, precultured in the presence of FCS, was assayed on the memory cytolytic T lymphocytes (CTL) response to alloantigens. For this, we have used in vitro conditions in which both particulate alloantigen and MLC SN are required to allow the generation of CTL. It was shown that the CTL response was totally inhibited in the presence of 5 to 7 days precultured spleen cells. This inhibitory effect was partly due to removal, by those precultured cells, of relevant factor(s) contained in the MLC SN. After velocity sedimentation at unit gravity, it was shown that the T cells able to inhibit the cytolytic response and to remove MLC SN factor(s) are found in the fractions containing the large proliferating cells. It was further demonstrated that in the presence of inhibiting cells, a significant CTL response may be obtained after addition of concentrated MLC SN. However, in this way, this inhibitory effect was not totally circumvented, which suggests that the memory CTL response is also impaired by other mechanisms.

[Trial of purification and serological characterization of human and murine transplantation antigens]. / Essai de purification et caractérisation sérologique des antigènes de transplantation humains et murins

Human and murine antigens are purified by exclusion chromatography on Sephadex and preparative polyacrylamide gel electrophoresis and their purification stage checked by analytical methods. The study of antigenic preparations consists of electrofocusing, molecular weight estimation and chemical determinations. A cytotoxicity inhibition test with specific alloantisera reveals the antigenic potency. A microtest using 51Cr labelled tumoral cells has been achieved for the analysis of murine preparations.

Direct in vitro inactivation of murine alloreactive cytotoxic-T-lymphocyte precursors by syngeneic nonspecific cytotoxic-T-cell populations.

Nonspecific cytotoxic T-cell populations, derived from murine fetal calf serum (FCS)-precultured cells expanded in interleukin 2 (IL-2)-containing supernatant (operationally defined as FCS-CM-expanded cells), were investigated for their inactivating properties on syngeneic lymphoid cell populations containing alloreactive cytotoxic-T-lymphocyte precursors (CTL-P). CTL-P were detected and quantitated in a limiting dilution mixed leukocyte microculture (micro-MLC) system supplemented with IL-2. The data show a dramatic decrease in relevant CTL-P frequency in populations of fresh or Day 2 in vitro-alloactivated peripheral blood leukocytes (PBL) after coculturing them for 24 hr with syngeneic mitomycin C-treated populations of FCS-CM-expanded cells. On the contrary, no decrease in CTL-P frequency was observed when Day 7, instead of fresh or Day 2, in vitro-alloactivated PBL were used as responding cells. Throughout these experiments, it was clearly shown that a decrease or an absence of CTL response in the micro-MLC was neither due to a lack of IL-2 nor to a premature destruction of the stimulating cells by the inhibiting FCS-CM-expanded cells still present in the culture. FCS-CM-expanded cells can destroy (in a 3-hr 51Cr-release assay) Day 2 alloreactive PBL populations, and this raises the possibility that the direct inactivation of CTL-P by FCS-CM-expanded cells could result from their cytolytic activities.

Generation of cytolytic T lymphocytes in vitro. XI. Accessory cell requirement in secondary responses to particulate alloantigen.

The non-T accessory cell requirement for cytolytic T lymphocyte (CTL) generation in vitro was studied in a system which makes use of particulate membrane preparations as a source of alloantigen, and spleen cells from alloimmune mice as a source of responding cells. It is shown that removal of nylon-adherent cells from the responding cell population strongly reduced CTL generation, whereas direct removal of Ig+, phagocytic or plastic-adherent cells had no effect. The CTL response of the nylon-nonadherent cell population could be reconstituted by the addition of normal spleen cells, which by themselves do not generate CTL in response to particulate alloantigen. The accessory cell function of normal spleen cells was not affected by depletion of T cells or of phagocytic cells, but was sensitive to gamma-irradiation (1000 rd). The system thus demonstrates the requirement for a nylon-adherent accessory cell population in the secondary CTL response to particulate alloantigens which does not exhibit the typical characteristics of T cells, B cells or macrophages.

[In vitro distinction between primary and secondary cellular response to alloantigens by means of a subcellular antigenic preparation]. / Distinction in vitro des réponses cellulaires primaire et secondaire contre les alloantigènes au moyen d'une préparation antigénique subcellulaire

Living spleen cells and a subcellular product of spleen cells have been compared, in mice, for their ability to elicit, for a long term culture (4-5 days), a cellular cytotoxic response against alloantigens. Contrary to living spleen cells which could induce the same high level of cytotoxic activity in a primary or in a secondary response, the antigenic preparation was only able to mount a very low primary response while it could render highly cytotoxic a lymphoid population proved to be able to mount in vitro an anamnestic response.

In vitro production of granular T cells and mast cells by use of different conditioned media. Ultrastructural and functional analysis.

Normal mouse spleen cells were cultured in different conditioned media (CM). Mixed lymphocyte culture supernatant (MLC SN) was shown to promote the proliferation of cytotoxic, Thy-1+, Lyt-1+, Lyt-2+, asialo-GM-1+, weakly adherent cells with numerous vacuoles and lysosome-like cytoplasmic granules. In contrast, the Con A SN induced the proliferation of non-cytotoxic, Thy-1-, Lyt-1-, Lyt-2-, asialo-GM-1-, non-adherent cells with numerous cytoplasmic granules. The ultrastructural morphology of these cells and the cytochemical characteristics of their granules enable us to identify them as mast cells. The different effects of both CM could be related to their T-cell growth factor (TCGF) content. When the amount of TCGF of these two CM was determined (by assaying growth-stimulating activity for T-cell colonies), it appeared that the MLC SN contained larger amounts of TCGF than the Con A SN used in these experiments.