The impact of C[O.sub.2] dry baths on subpopulations of NK and NK-T lymphocytes, cytotoxic activity, level of inflammation and pain management in elderly with musculosceletal syndromes--a pilot study.Introduction
The use of carbonated water in the form of carbon dioxide carbon dioxide, chemical compound, CO2, a colorless, odorless, tasteless gas that is about one and one-half times as dense as air under ordinary conditions of temperature and pressure. baths is based on traditional therapeutic methods and is currently limited in the area of "Medical Spa treatments" or "Balneo-therapeutics". C[O.sub.2] treatment can be applied with two different ways; with carbon dioxide enriched water and with dry carbon dioxide bath (DCDB DCDB Domain Control DataBase
DCDB Digital Cities Data Base
DCDB Domain Control Data Base
DCDB Detector Construction Data Base ). It has to be mentioned that there are essential differences between the two C[O.sub.2] bath methods, which differentiates them completely. In carbon dioxide bath (CDB CDB Common Database
CDB Caribbean Development Bank
CDB Convention sur la Diversité Biologique (Convention on Biological Diversity)
CDB China Development Bank (Beijing, China)
CDB Capital Development Board ), C[O.sub.2] gets in contact with the human body via its dilution in water and in this way it is possible for an individual to inhale the gas. In addition, the immersed body is affected by water buoyancy, hydrostatic pressure and water temperature. In DCDB, the body comes in contact only with C[O.sub.2]. Most studies focus more on the effect of CDB and less of DCDB. DCDB is thought to be effective mainly in the treatment of peripheral vascular diseases, however, the mechanism(s) underlying this traditional therapy remain(s) poorly defined.
In a recent study of 2009, Gapon and Ignatov reported that the therapy with the use of DCDB after acute myocardial infarction acute myocardial infarction (·kyōōtˑ mī·ō·karˑ·dē· , decreases systolic Systolic
The phase of blood circulation in which the heart's pumping chambers (ventricles) are actively pumping blood. The ventricles are squeezing (contracting) forcefully, and the pressure against the walls of the arteries is at its highest. and diastolic Diastolic
The phase of blood circulation in which the heart's pumping chambers (ventricles) are being filled with blood. During this phase, the ventricles are at their most relaxed, and the pressure against the walls of the arteries is at its lowest. arterial pressure and generally reduces cardiovascular complications. (1) Seven years earlier, Abramovich had observed that in elderly patients with hypertension dry carbon dioxide baths rehabilitate skin thermoreactivity and improve microcirculation microcirculation /mi·cro·cir·cu·la·tion/ (-sir?ku-la´shun) the flow of blood through the fine vessels (arterioles, capillaries, and venules).microcirculato´ry
n. . (2) Decrease of arterial pressure is mentioned in other studies also. (3,4) The application of DCDB in patients with psoriatic arthritis Psoriatic Arthritis Definition
Psoriatic arthritis is a form of arthritic joint disease associated with the chronic skin scaling and fingernail changes seen in psoriasis. was directly associated with the positive results in immunological markers. (5) Grigor'eva et al report a 70% clinical improvement with the combined use of dry-air carbon dioxide baths and applications of peloids at low temperatures in the rehabilitation of 106 patients with psoriatic arthritis. (6)
The aim of this study is to investigate the efficiency of treatment with anhydrous an·hy·drous
Without water, especially water of crystallization.
adj without water.
containing no water. C[O.sub.2] for hypothetical immune boosting actions (according to traditional data) in elderly patients suffering from painful musculoskeletal musculoskeletal /mus·cu·lo·skel·e·tal/ (-skel´e-t'l) pertaining to or comprising the skeleton and muscles.
Relating to or involving the muscles and the skeleton. syndromes. Secondary aims of this research are to record in this category of patients biochemical parameters of inflammation, possible haemodynamic responses, pain perception and mood changes.
For the assessment of hemodynamic responses, we examined the arterial pressure, heart rate and body temperature before and after the intervention in every session. To assess immunological responses we measured white blood cells White blood cells
A group of several cell types that occur in the bloodstream and are essential for a properly functioning immune system.
Mentioned in: Abscess Incision & Drainage, Bone Marrow Transplantation, Complement Deficiencies , lymphocytes, natural killer cells natural killer cells,
n.pl lymphocytes that are part of innate immunity that kill foreign substances and abnormal tissues. Decreased number or activi-ty has been linked to a number of diseases, including AIDS, cancer, chronic fatigue syndrome, (NK), natural killer T cells (NK-T), and cytotoxicity, at the same times. For investigating the pain perception and mood we measured the endogenous opioid peptide opioid peptide Endogenous opiate Any natural polypeptide neurotransmitters involved in perception of pain, response to stress, regulation of appetite, sleep, memory, learning , [beta]-endorphin. And for the markers of inflammation we measured tachykinins, substance P and neurokinin A.
Evidence based data on the correlation and interaction between measured variables
Though the impact of substance P and neurokinin A on the immune system immune system
Cells, cell products, organs, and structures of the body involved in the detection and destruction of foreign invaders, such as bacteria, viruses, and cancer cells. Immunity is based on the system's ability to launch a defense against such invaders. remains unclear, many studies indicate that, there may also be exerted profound influence on inflammatory responses, by affecting multiple aspects of immune cell function. (7) This fact establishes a receptor-mediated mechanism by which tachykinin peptides can affect both innate and specific immune responses. (8) Substance P was shown to induce and mediate inflammation, angiogenesis angiogenesis /an·gio·gen·e·sis/ (-jen´e-sis) vasculogenesis; development of blood vessels either in the embryo or in the form of neovascularization or revascularization.
n. , infections, intestinal mucosal immunity and stress and also activate several immune cells, such as CD4+ and CD8+ T lymphocytes, mast cells Mast cells
A type of immune system cell that is found in the lining of the nasal passages and eyelids, displays a type of antibody called immunoglobulin type E (IgE) on its cell surface, and participates in the allergic response by releasing histamine from , NK cells and macrophages Macrophages
White blood cells whose job is to destroy invading microorganisms. Listeria monocytogenes avoids being killed and can multiply within the macrophage. . (9) The functionally active neurokinin-1 receptors can be expressed by human natural killer cells. Substance P might therefore be a novel link between neural structures and innate immunity. (10)
Material and methods
In this research, 37 patients suffering from chronic musculoskeletal syndromes participated. They were randomly divided into two groups. In group A, 27 people participated with an average age of 64,3 [+ or -] 5,67 y (12 men with an average age of 67,7 [+ or -] 5,69 y, 15 women with an average age of 62,9 [+ or -] 5,05 y), and in group B (placebo), 10 people participated with an average age of 65,1 [+ or -] 5,33y (5 men with an average age of 66,4[+ or -]5,23y, 5 women with an average age of 64,4 [+ or -] 5,96y). The participants did not discontinue any pharmaceutical prescription nor did they change it at any point during the experiment. Their diet and routine habits were not altered in any way. During the intervention, one individual was disqualified because he deviated from the prescribed administration of drugs.
Eligibility criteria: age range from 60 to 70 years, Musculoskeletal problem, hypertension, reported generalized weakness, and bad mood (not related to bipolar disorder).
The selected individuals had at least one health problem, from each category listed below:
1. Cardiovascular disease: hypertension, chronic heart failure, coronary heart disease coronary heart disease: see coronary artery disease.
coronary heart disease
or ischemic heart disease
Progressive reduction of blood supply to the heart muscle due to narrowing or blocking of a coronary artery (see atherosclerosis). , chronic peripheral vascular disease of the lower extremities.
2. Painful musculoskeletal syndromes: chronic low back pain, lumbar disc herniation herniation /her·ni·a·tion/ (her?ne-a´shun) abnormal protrusion of an organ or other body structure through a defect or natural opening in a covering, membrane, muscle, or bone. , cervical disc herniation, neck syndrome, osteoarthritis osteoarthritis
or osteoarthrosis or degenerative joint disease
Most common joint disorder, afflicting over 80% of those who reach age 70. It does not involve excessive inflammation and may have no symptoms, especially at first. , shoulder periarthritis, tendonitis tendonitis /ten·do·ni·tis/ (ten?do-ni´tis) tendinitis.
Variant of tendinitis. .
3. Osteoporosis with chronic pain (all women)
4. Diabetes mellitus (noticed on one individual).
The anthropometric an·thro·pom·e·try
The study of human body measurement for use in anthropological classification and comparison.
an characteristics were as follows:
Body weight (kg): The mean weight (Kg) of all patients was 82,6 [+ or -] 13,25, for men 92,8 [+ or -] 6,77, for women 74,5 [+ or -] 11,47 with no significance (p) of mean differences between men and women (sex).
Height (m): mean value 1,65 [+ or -] 6,7, for men 1,74 [+ or -] 1, for women 1,61 [+ or -] 3,2 and the significance (p) of mean differences according to sex was p<0.0001. Body Mass Index (BMI BMI body mass index.
body mass index
Body mass index (BMI)
A measurement that has replaced weight as the preferred determinant of obesity. ): mean value 29,01 [+ or -] 3,63, for men the mean value was 30,07 [+ or -] 2,9, for women mean value was 28,56 [+ or -] 4,01, and there was not significance (p) of mean differences between the two sexes.
C[O.sub.2] dry bath
This method requires a special plastic bag made of hypoallergic material, used to cover each participant's naked body. The plastic bag is sealed with tape on the chest or neck level. Then the individual lies on a bed in a quiet room. Dry C[O.sub.2] is very slowly administered into the bag through a tube, until the bag gets inflated with 180gr of C[O.sub.2]. C[O.sub.2] is heavier than atmospheric air. The time required for this session is 60 minutes. After this, red pigmentation pigmentation, name for the coloring matter found in certain plant and animal cells and for the color produced thereby. Pigmentation occurs in nearly all living organisms. appears on the skin due to the action of C[O.sub.2] on skin capillaries (vasodilatation vasodilatation /vaso·di·la·ta·tion/ (-di?lah-ta´shun) vasodilation.
a state of increased caliber of blood vessels. ). The number of sessions were 17 in total.
Design of the study
Participants who volunteered in this project had no food or water intake during their daily therapy sessions. Their height, weight and index of body mass were measured. They all underwent a clinical examination to determine the general condition of their health. This is indicated with a "good health" report sheet. It should be noted that the participants' weight was measured and recorded in the morning by an accurate balance (Electronil, Soehnle, West Germany). Their height was measured using a seca height measurement device (seca 240 Telescopic Height Measure, Germany). Furthermore, their blood pressure was measured prior to and following each DCDB using an Hg manometer (Baum, Baumanometer, Desk Model, USA).
Blood Samples (collection-distribution)
10ml of whole blood was drawn carefully in the morning from the median cubital vein In human anatomy, the median cubital vein (or median basilic vein) a superficial vein of the upper limb. It connects the basilic and cephalic vein and is often used for venipuncture (taking blood). It lies in the cubital fossa superficial to the bicipital aponeurosis. of each participant. For sampling, BD Vacutainer[R] Blood Collection Tubes were used. The blood was distributed equally (5ml) in tubes containing anticoagulant anticoagulant (ăn'tēkōăg`yələnt), any of several substances that inhibit blood clot formation (see blood clotting). (Heparin) and in others without it. The blood was centrifuged in 2000 rpm for 5 minutes (Biofuge, 17RS, Heraeus, SEPATECH, Gmb, Osterode, Denmark). The serum was collected and stored in -800[degrees]C for 6 weeks.
Methodology of NK cytotoxicity by flow cytometry (FCA FCA
Abbreviation for the Free Carrier )
The basic principle of the quantification of the cytotoxic activity of NK cells with FCA is to discriminate between effector effector /ef·fec·tor/ (e-fek´ter)
1. an agent that mediates a specific effect.
2. an organ that produces an effect in response to nerve stimulation. (NK cells) and target (cancer) cell populations. The cell line K562 (cryopreserved) is used as target cells prestained with green fluorescent membrane dye (Beckton-Dickinson, USA). The K562 cell line derived from the blood of a patient suffering from chronic myeloid leukemia in terminal blastic crisis, and represents the most sensitive target cell line for human NK cells. (11) K562 cells lack MHC MHC major histocompatibility complex.
major histocompatibility complex
major histocompatibility complex. classes I and II antigens. After incubation of the effector and the target cells, a red fluorescent DNA DNA: see nucleic acid.
or deoxyribonucleic acid
One of two types of nucleic acid (the other is RNA); a complex organic compound found in all living cells and many viruses. It is the chemical substance of genes. dye (Sigma, USA) is added to label the target cells permeabilized by NK activity. This dye labels only cells with compromised plasma membranes. In this way, a clear separation between four cell populations can be obtained: live target cells, dead target cells, live effector cells, and dead effector cells. Thus, the actual ratio between effector and target cells (E:T) can be confirmed, but only events that appear to be positive after this analysis (dead and live targets cells) will have to be collected for the determination of the NK cytotoxic activity. This method described by Kane et al (11) included four steps: a) isolation of effector cells includes peripheral blood mononuclear mononuclear /mono·nu·cle·ar/ (-noo´kle-er)
1. having but one nucleus.
2. a cell having a single nucleus, especially a monocyte of the blood or tissues.
adj. cells (PBMCs) containing the NK cells, b) thawing of K562 target cells, c) cytotoxicity assay, and d) flow cytometric analysis (Figure 1).
Methodology of estimation of the number of NK and NK-T cells
For the estimation of the populations of NK and NK-T cells the technique of flow cytometry (Becton-Dickinson, USA) was used.
The Elisa technique (Phoenix Pharmaceutical, Inc, Germany) was used to identify the substances [beta]-endorphin (ng/ml), substance P (ng/ml) and neurokinin A (ng/ml) in human serum. An Elisa Reader was used to measure the absorption of the above peptides (Expert Plus UV, Microplate Reader, ASYS Hitech GmbH Eugendorf, Austria).
All data was analyzed using SPSS A statistical package from SPSS, Inc., Chicago (www.spss.com) that runs on PCs, most mainframes and minis and is used extensively in marketing research. It provides over 50 statistical processes, including regression analysis, correlation and analysis of variance. 14.0 for Windows. Appropriate descriptive statistics (mean and standard deviation for all measurable variables) and the parametric method Student's t test (independent-samples t test, 2-tailed) were used for the statistical analysis. The one-sample Kolmogorov-Smirnov test was used to examine whether the data of each of the variables, entering the Student's t-test analysis, were normally distributed. Our data were assessed using the Pearson's Correlation technique. The level of significance was set at 0.05.
In table 1, the measurements of body temperature and cardiac parameters are presented. Men had higher systolic blood pressure Systolic blood pressure
Blood pressure when the heart contracts (beats).
Mentioned in: Hypertension (mmHg) than women before (p=0.010) and after (0.002) the intervention. In women the diastolic arterial pressure was decreased (p=0.004) after the intervention. The pulse rate had a significant decrease (p=0.0001) after the DCDB in all participants and no difference was noted between men and women. It should be noticed that the women's heart rate was higher (p=0.007) than the men's in the measurements before the sessions.
Significant increases of [beta]-endorphin levels were observed in all participants after the sessions. Similar increases were found in the Substance P. Neurokinin A levels were increased in the total sample and in men also (Table 2).
The white cells count was increased only in men. The lymphocytes count (%) was increased only in women, who had higher levels before and after the sessions. The levels (%) of NK cells increased in men (p=0.039). The NK-T cells increased in all participants (total, men and women) and presented significant (p=0.002) differences according to sex before the sessions. Improvement of cytotoxicity resulted after the sessions (Table 3).
[FIGURE 1 OMITTED]
In the present study the measured outcomes derived from the action of dry C[O.sub.2] in the human organism, with actions made via the skin and the open body cavities, anus and urethra urethra (yrē`thrə), canal in most mammals that carries urine from the bladder to the outside of the body; in the male it also serves as a genital duct. . Several researchers showed that water immersion, especially in C[O.sub.2]-enriched water, induces bradycardia bradycardia: see arrhythmia. via vagus nerve stimulation vagus nerve stimulation Psychiatry Electroconvulsive therapy
in which a pacemaker-like device stimulates the vagus nerve. See Electroconvulsive therapy. and the absorption of C[O.sub.2] from the skin surface (12), it normalizes the heart rhythm (13) or acts on the subcutaneous microcirculation by increasing parasympathetic parasympathetic /para·sym·pa·thet·ic/ (-sim?pah-thet´ik) see under system.
Of, relating to, or affecting the parasympathetic nervous system. and decreasing sympathetic activity. (14,15) In our research the absence of water and high temperature, proves that the considerable reduction of heart rate is due to the direct action of C[O.sub.2] to the parasympathetic system. This is explained by the increase of neuropeptides neuropeptides (ner·ō·pepˑ·tīdz),
n.pl endogenous protein molecules that influence neural activity by carrying information directly to the cells and tissues. that suppress the actions of sympathetic system.
The synchronous increase of neuropeptides and NK cells agrees with many studies in the area of neuroimmunomodulation research (16,17) and the finding that there is secretion of opioid peptides by leukocytes. (18-22) In addition, endorphins endorphins (ĕndôr`fĭnz), neurotransmitters found in the brain that have pain-relieving properties similar to morphine. There are three major types of endorphins: beta endorpins, found primarily in the pituitary gland; and enkephalins and and substance P increase NK cell cytotoxicity. (23)
The increase of [beta]-endorphin levels explains the pleasant disposal and well-being that was described by all participants. The interactions between leukocyte-derived opioid peptides and peripheral nociceptor nociceptor /no·ci·cep·tor/ (-sep´ter) a receptor for pain caused by injury, physical or chemical, to body tissues.nocicep´tive
A sensory receptor that responds to pain. endings carrying opioid receptors (24,25) may explain the connection of the pathophysiologic common basis of all the examined variables in our study with the pain mechanisms. The presence of hypodermic hypodermic /hy·po·der·mic/ (-der´mik) applied or administered beneath the skin.
1. Of or relating to the layer just beneath the epidermis.
2. receptors of substance P, neurokinin A (26), [beta]-endorphin (26,27) and their stimulation by C[O.sub.2] explains that endogenous opioid analgesia analgesia /an·al·ge·sia/ (an?al-je´ze-ah)
1. absence of sensibility to pain.
2. the relief of pain without loss of consciousness. results from neuroimmune interactions that occur in peripheral tissues. (28)
What should also be highlighted is that the level of sensitivity of C[O.sub.2] receptors that have been recognised in mosquitoes, explain how easily these insects can detect low levels of C[O.sub.2] in humans. (29) Finally, because oxygen and carbon dioxide are very important for the function of the human organism, numerous chemosensitive neurons can either obstruct or facilitate openings for stimulations, depending on the varying conditions. (30) Our study incicates that C[O.sub.2] affects the neuropeptides and their receptors which are located on the skin. These skin receptors are as important as other receptors found in the lungs, brain and other vital organs. These receptors rapidly allow the organism to conceptualize environmental alterations in order to adapt. Frequent or continuous contact of C[O.sub.2] to the skin seems to be perceived by the organism as an unpleasant situation.
In conclusion, C[O.sub.2] via the DCDB technique seems to have immune boosting results in elderly patients with chronic musculoskeletal syndromes. Other results of this treatment is decrease of hypertension, pain management and euphoric sensation. Those results are short--term responses that have been measured after the C[O.sub.2] sessions. Further investigation will be needed to determine the long term effects of C[O.sub.2]. Also, the exact participation of the skin receptors in the pathophysiologic mechanisms of the neuroimmune post session response are to be defined with skin biopsies.
We would like to express our thanks to the "Physiokinisi" Company and especially to Ms. Renata Mihailidou for the support.
Conflict of interest: None declared.
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The processes by which many animals actively maintain the temperature of part or all of their body within a specified range in order to stabilize or optimize temperature-sensitive physiological processes. in hypertensive hypertensive /hy·per·ten·sive/ (-ten´siv)
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Elias Kallistratos , Ioannis Toliopoulos , Dimitrios Fragkoraptis , Spyridon Gerou , Eleftherios Fragkoraptis 
 Physiotherapy Department, Alexander Technological Educational Institute, Thessaloniki, Greece,
 Konstantinion Research Center of Molecular Medicine and Biotechnology, Thessaloniki, Greece
 Analysis Research and Diagnostic Biomedical bi·o·med·i·cal
1. Of or relating to biomedicine.
2. Of, relating to, or involving biological, medical, and physical sciences. Labs, Greece
Elias Kallistratos, Ph.D, Associate Professor Physiotherapy Department, Alexander Technological Institute of Thessaloniki P.O BOX 141, GR-57400 Sindos, Thessaloniki, Greece Tel: +30 2310 791548, Mobile: +30 6972405216 E-mail: firstname.lastname@example.org
Table 1. Measurements of Body temperature and cardiac parameters. Parameters Total Total T-test (P) T.b ([degrees]C) 36.5 [+ or -] 0.03 T.a ([degrees]C) 36.5 [+ or -] 0.06 NS BPs.b (mmHg) 133.0 [+ or -] 12.97 BPs.a (mmHg) 131.4 [+ or -] 12.45 NS BPd.b (mmHg) 82.2 [+ or -] 6.50 BPd.a (mmHg) 80.9 [+ or -] 7.40 NS P.b (BPM) 75.1 [+ or -] 7.80 P.a (BPM) 68.9 [+ or -] 6.26 0.000 Parameters Men Men T-test (P) T.b ([degrees]C) 36.5 [+ or -] 0.02 T.a ([degrees]C) 36.5 [+ or -] 0.01 NS BPs.b (mmHg) 139.9 [+ or -] 4.96 BPs.a (mmHg) 139.1 [+ or -] 5.02 NS BPd.b (mmHg) 81.8 [+ or -] 6.11 BPd.a (mmHg) 82.4 [+ or -] 8.30 NS P.b (BPM) 70.8 [+ or -] 5.92 P.a (BPM) 66.8 [+ or -] 6.22 0.000 Parameters Women Women T-test (P) T.b ([degrees]C) 36.5 [+ or -] 0.03 T.a ([degrees]C) 36.5 [+ or -] 0.08 NS BPs.b (mmHg) 127.5 [+ or -] 14.83 BPs.a (mmHg) 125.2 [+ or -] 13.28 NS BPd.b (mmHg) 82.5 [+ or -] 6.99 BPd.a (mmHg) 79.8 [+ or -] 6.67 0.004 P.b (BPM) 78.5 [+ or -] 7.54 P.a (BPM) 70.6 [+ or -] 5.98 0.000 Parameters Men-Women T-test T.b ([degrees]C) 0.014 T.a ([degrees]C) ND BPs.b (mmHg) 0.010 BPs.a (mmHg) 0.002 BPd.b (mmHg) NS BPd.a (mmHg) NS P.b (BPM) 0.007 P.a (BPM) NS T.b. (Body Temperature before), T.a. (Body Temperature after), BPs.b (systolic blood pressure before), BPs.a (systolic blood pressure after), BPd.b (diastolic blood pressure before), BPd.a (diastolic blood pressure after), P.b. (pulse rate before), P.a. (pulse rate after), NS: no significant difference Table 2. The levels of the peptides before (b) and after (a) the protocol Parameters Total T-test (P) Men ng/ml B-endorphin.b 1.1 [+ or -] 0.57 1.3 [+ or -] 0.65 B-endorphin.o 2.8 [+ or -] 2.26 0.001 3.9 [+ or -] 2.89 Substance P.b 0.7 [+ or -] 0.18 0.7 [+ or -] 0.09 Substance P.a 0.9 [+ or -] 0.33 0.002 0.9 [+ or -] 0.3 Neurokinin A.b 3.1 [+ or -] 0.6 3 [+ or -] 0.48 Neurokinin A.a 3.5 [+ or -] 0.72 0.041 3.50.62 Parameters T-test (P) Women ng/ml B-endorphin.b 1 [+ or -] 0.49 B-endorphin.o 0.024 1.9 [+ or -] 1.03 Substance P.b 0.6 [+ or -] 0.21 Substance P.a 0.005 0.9 [+ or -] 0.36 Neurokinin A.b 3.1 [+ or -] 0.7 Neurokinin A.a 0.023 3.4 [+ or -] 0.81 Parameters T-test (P) T-test ng/ml differences Men-Women B-endorphin.b NS B-endorphin.o 0.000 0.021 Substance P.b NS Substance P.a 0.046 NS Neurokinin A.b NS Neurokinin A.a NS NS NS: no significant difference Table 3. Measurements of immune system markers Parameters Total T-test (P) HCT.b. (%) 41.8 [+ or -] 6.05 HCT.a. 43.2 [+ or -] 5.34 0.000 WBC.6.103/uL 6 [+ or -] 0.9 WBC.a. 5.6 [+ or -] 0.95 0.016 Lymp.b. % 30.7 [+ or -] 7.8 Lymp.a 33.4 [+ or -] 8.99 0.001 NK.b. % 14,1 [+ or -] 4,96 NK.a. 17,4 [+ or -] 5,77 NS NK-T.b. % 11,1 [+ or -] 6,16 NK-T.a. 3,3 [+ or -] 1,23 1 NK-T12.5:1.b. 5,7 [+ or -] 2,18 NK-T12.5:1.a. 20 [+ or -] 12,48 0.000 NK-T25:1.b. 7,3 [+ or -] 1,86 NK-T25:1.a 21,8 [+ or -] 12,01 0.000 NK-T50:1.b. 12,4 [+ or -] 3,89 NK-T50:1.a. 36,4 [+ or -] 13,45 0.000 Parameters Men T-test (P) HCT.b. (%) 44.5 [+ or -] 7.64 HCT.a. 45.9 [+ or -] 6.95 0.002 WBC.6.103/uL 6.1 [+ or -] 0.35 WBC.a. 5.3 [+ or -] 0.51 0.000 Lymp.b. % 25.2 [+ or -] 5.76 Lymp.a 25.4 [+ or -] 5.37 NS NK.b. % 13,2 [+ or -] 2,57 NK.a. 20,2 [+ or -] 5,7 39 NK-T.b. % 7,3 [+ or -] 2,22 NK-T.a. 3,3 [+ or -] 0,47 1 NK-T12.5:1.b. 7,2 [+ or -] 2,2 NK-T12.5:1.a. 24,1 [+ or -] 16,14 24 NK-T25:1.b. 8,4 [+ or -] 1,83 NK-T25:1.a 21,8 [+ or -] 15,8 12 NK-T50:1.b. 13,4 [+ or -] 4,48 NK-T50:1.a. 43,7 [+ or -] 15,15 0.000 Parameters Women T-test (P) HCT.b. (%) 39.6 [+ or -] 3.23 HCT.a. 41.1 [+ or -] 2.03 0.002 WBC.6.103/uL 6 [+ or -] 1.19 WBC.a. 6 [+ or -] 1.12 NS Lymp.b. % 35.2 [+ or -] 6.27 Lymp.a 39.7 [+ or -] 5.46 0.000 NK.b. % 14,7 [+ or -] 6,28 NK.a. 15,2 [+ or -] 5,1 NS NK-T.b. % 14,1 [+ or -] 6,66 NK-T.a. 3,3 [+ or -] 1,64 0.000 NK-T12.5:1.b. 4,6 [+ or -] 1,33 NK-T12.5:1.a. 16,7 [+ or -] 7,69 0.000 NK-T25:1.b. 6,5 [+ or -] 1,41 NK-T25:1.a 21,8 [+ or -] 8,47 0.000 NK-T50:1.b. 11,6 [+ or -] 3,26 NK-T50:1.a. 30,6 [+ or -] 8,61 0.000 Parameters T-test differences Men-Women HCT.b. (%) 0.031 HCT.a. 0.017 WBC.6.103/uL NS WBC.a. NS Lymp.b. % 0.000 Lymp.a 0.000 NK.b. % NS NK.a. NS NK-T.b. % 2 NK-T.a. NS NK-T12.5:1.b. 1 NK-T12.5:1.a. NS NK-T25:1.b. 5 NK-T25:1.a NS NK-T50:1.b. NS NK-T50:1.a. 0,009 b: before, a: after, HCT: Heamatocrit, WBC: White blood cells, Lymp.: lymphocytes, NK NK cells, NK-T: NK-T cells, NK-T 12.5:1, 25:1 & 50:1 cytotoxicity of NK cells in 3 ratios, NS: no significant difference