Revista Română de Medicină de Laborator Vol. 24, Nr.

3, Septembrie, 2016 327

Original research article

DOI: 10.1515/rrlm-2016-0008

Biochemical and functional modifications in biathlon
athletes at medium altitude training
Modificările biochimice și funcționale ale atleților biatloniști după
antrenament la altitudine medie

Dana Bădău1, Anca Bacârea2,*, Ramona Natalia Ungur3, Adela Bădău3, Alina
Mirela Martoma4
Department of Human Movement Sciences, University of Medicine and Pharmacy Tîrgu Mures, Romania;
Department of Pathophysiology, Univeristy of Medicine and Pharmacy Tîrgu Mureș, Romania;
Department of Physical Education, University of Medicine and Pharmacy Tîrgu Mures, Romania;
Department of Physical Education and Special Motricity, Transilvania University of Brasov, Romania

Objective: The aim of our research was to identify physiological and biochemical changes induced by training at
medium altitude.
Methods: Ten biathlon athletes underwent 28-day training camp at medium altitude in order to improve their
aerobic effort, following the living high-base train high-interval train low (Hi-Hi-Lo) protocol. There were investi-
gated three categories of functional and biochemical parameters, targeting the hematological changes (RBC, HCT,
HGB), the oxidative (lipoperoxid, free malondialdehyde and total malondialdehyde) and antioxidative balance (the
hydrogen donor capacity, ceruloplasmin and uric acid) and the capacity of effort (the maximum aerobic power, the
cardiovascular economy in effort, the maximum O2 consumption).
Results: All the biochemical and functional evaluated parameters showed significant increases between the
pre-training testing and post-training testing (5.13 ± 0.11 vs. 6.50 ± 0.09, p < 0.0001 for RBC; 44.80 ± 1.22
vs. 51.31 ± 2.31, p < 0.0001 for HCT; 15.06 ± 0.33 vs. 17.14 ± 0.25, p < 0.0001 for HGB; 1.32 ± 0.04 vs.1.62
± 0.01, p < 0.0001 for LPx; 1.61 ± 0.01 vs. 1.73 ± 0.01, p < 0.0001 for free MDA; 2.98 ± 0.08 vs. 3.37 ± 0.03,
p  < 0.0001 for total MDA; 45.92 ± 0.13 vs. 57.98 ± 0.12, p < 0.0001 for HD; 25.95 ± 0.13 vs. 31.04 ± 0.06,
p < 0.0001 for Crp; 3.47 ± 0.03 vs.7.69 ± 0.02, p < 0.0001 for UA; 63.91 ± 1.00 vs. 81.53 ± 1.97, p < 0.0001 for
MAP; 33.13 ± 0.57 vs. 57.41 ± 0.63, p < 0.0001 for CVEE; 4190 ± 50.45 vs. 5945 ± 46.48, p < 0.0001 for VO2max).
Conclusions: Aerobic effort capacity of biathlon athletes has increased in the post-training period, using Hi-Hi-Lo
Keywords: acclimatization, medium altitude, hematological and functional explorations, biathlon, Hi-Hi-Lo

* Corresponding author: Anca Bacârea, Department of Pathophysiology, Univeristy of Medicine and Pharmacy
Tîrgu Mureș, Str. Gh. Marinescu, Nr. 5, Tîrgu Mureș, Romania. e-mail:

01 vs. effort intensity mmHg.0001 for HGB. in case chemical changes in the body’s systems.08 vs. 1. concentration [5]. explorări hematologice și funcționale.95 ± 0.69 ± 0. These adaptations in- crease.0001 for free MDA.04 ± 0.63.03. 5945 ± 46.1. 2. balanța oxidativă (li- poperoxid. HGB). urmând protocolul (Hi-Hi-Lo). These procedures have in altitude implies a proportional increase in the been adopted by biathletes and cross-country intensity of hyperventilation.91 ± 1. p < 0. de- gets into hyperventilation to ensure the optimum termining the improvement of the effort capaci- O2 need for body acclimatization. 31.13 ± 0. 4190 ± 50. 63.92 ± 0. capillary density and tissue myoglobin values. ven- m altitude. 17. în scopul de a îmbunătăți efortul aerob.7.62 ± 0.328 Revista Română de Medicină de Laborator Vol. p < 0. 2016 Rezumat Obiectiv: Scopul cercetării noastre a fost de a identificarea modificărilor fiziologice și biochimice induse de antre- namentul la altitudine medie.0001 for MAP. both the oxygen transportation and are essential.0001 for Crp. 1. the athletes’ bodies adapt by Body acclimatization to alpine climate with increasing the RBC rate formation and by the medium altitude induces physiological and bio- rapid mobilization of the blood reserves.0001 for LPx. 1. ceruloplasmina și acidul uric) și capacitatea de efort (puterea maximă aerobă. 57. It has been estab- for survival [1-3]. 44.0001 for HCT.25. p < 0. 51. Septembrie. p < 0. Cuvinte cheie: aclimatizare. economia cardiovasculară în efort. Au au fost investigați trei categorii de para- metri funcționali și biochimici care vizează modificările hematologice (RBC. protocolul Hi-Hi-Lo.61 ± 0. 3.0001 for HD.0001 for RBC. p < 0. skiers [4].41 ± 0.31 ± 2. 15. la altitudine medie. 57.0001 for total MDA.47 ± 0.0001 for UA. . Nr. consumul maxim de O2).06 ± 0. At 1000 m altitude the human body the oxygen consumption capacity increase. p < 0.11 vs. Published: 30th August 2016. p < 0. 3. p < 0.73 ± 0.000.13 vs.02.50 ± 0. Introduction After a period of 2-4 weeks training and ex- posure at altitude. among they descend to a lower altitude where the pO2 which the circulatory and respiratory systems is normal. Material și metodă: Zece sportivi de biatlon au participat la antrenament timp de 28 zile.01.00 vs. The human body can compensate for the These adaptations are modulated by many lack of O2 up to an altitude of 5000 m.80 ± 1.000/mm3 at 500 clude the increase in HGB concentration. 25. 81. malondialdehida liberă și malondialdehida totală) și antioxidativă (capacitatea de donor de hidrogen. folosind protocolul Hi-Hi-Lo. Rezultate: Toți parametrii biochimici și funcționali evaluați au prezentat creșteri semnificative între testarea pre-antrenament și testarea post-antrenament (5. reaching values of 8. Accepted: 24th August 2016. p < 0. p  < 0.14 ± 0.33 vs.06. 24.48. where factors. lished that exposure to high altitude is an en- The body has mechanisms to prevent tissue vironmental stressor that elicits a response that hypoxia by changing blood parameters.13 ± 0. p < 0.45 vs. the increase ty on short or long term. 45.57 vs.37 ± 0. 3. time of exposure.98 ± 0. values that represent the minimum limit and individual conditions. Received: 11th October 2015.31.0001 for CVEE. Concluzii: Capacitatea de efort aerob a sportivilor de biatlon a crescut în perioada post-antrenament.98 ± 0. contributes to many adjustments and adapta- at high altitude the hemoglobin concentration tions that influence the effort capacity and the (HGB) and the red blood cell count (RBC) in- endurance performance.03 vs. 6.22 vs. including the degree of hypoxia relat- the oxygen partial pressure (pO2) reaches 25-35 ed to altitude. biatlon.0001 for VO2max).32 ± 0. resulting in the doubling of normal tilation. HCT.13 vs. 33.09. p < 0.12. thus.53 ± vs. altitudine medie.

arterial blood at medium altitude exceeds 90% in the sports center of Cheile Gradistei . changes that will determine an tion of the hematological parameters. were volunteers and gave their informed consent. All the selected participants in this study cal performance. suffers a series of functional and biochemical The study comprised three categories of bio- changes in the case of exposure and training at chemical and functional explorations: the explora- medium altitude. 2. tion (HGB g/dl).1. pre-training and letes. lyze the hematological changes in relation with The athletes underwent the pre-training test- the effort or the motor capacity. ing prior to the start of exposure to altitude and In setting the research hypothesis we start.500-5. • For the antioxidative balance: the hydro- ducted in 2014 was achieved after three weeks of gen donor capacity (HD%). recorded. the exception that the post-training testing con. the hema- the oxidative and antioxidative balance parame. tocrit (HCT %) and the hemoglobin concentra- ters and the effort capacity parameters. The present research was conducted on a vided into three zones according to their altitude group of 10 experienced male biathlon athletes. .300 m. all expressed the same training program and protocol. 28 days after the expo- ed from the assumption that the athlete’s body sure to altitude. above the sea level [6]: who underwent 28-day training camp at medium 1.300 m. 2016 329 The high altitude environment may be di. and we tar- center of Rasnov at an altitude of 676 m and geted the following markers: only changes in the functional capacity were • For the oxidative balance: lipoperoxid (LPx).Fundata and tissue oxygenation is not constrained [7]. red blood cells count (RBC x106/µl). 3.500 m. This research is an extension of a previous For the determination of the oxidative and research performed between 15th of July and antioxidative balance we collected venous blood 11th of August 2014 on the same group of ath.over 5. with in nmol/ml. the determi- improvement of aerobic effort capacity. medium altitude . extreme altitude . but they do not fully highlight the The study was approved by the Ethics Commit- correlation between the adaptive changes of the tee of University of Medicine and Pharmacy Tîrgu human body and the improvement of the physi. ceruloplasmin physical training camp. the post-training testing. 3. nation of the oxidative and antioxidative balance The aim of our study is the assessment at me. dium altitude of three categories of parameters: The targeted hematological indicators were: the adaptive changes of the blood parameters. higher altitude . samples. when the training took place at the sports post-acclimatization.2. numerous. The hematological determina- tions were made using the Celldyn 3700 Abott Material and method analyzer. at an altitude of over 1500 m and in Postavarul Studies on adaptation to altitude are relatively Mountain range at an altitude of over 1700 m. Septembrie. Revista Română de Medicină de Laborator Vol. They followed the living high-base train high-in- terval train low (Hi-Hi-Lo) protocol. altitude in order to improve the aerobic effort.500-2. Mureș. Most studies particularly ana. The exposure to hypoxia and training was The saturation with oxygen (SaO2) of the done between 13th of July and 9th of August 2015. (Crp mg/dl) and uric acid (UA mg/dl). and the exploration of the capacity of effort. Nr. post-training. 24. pre-acclimatization. free malondialdehyde (MDAfree) and total Both studies were carried out by including malondialdehyde (MDAtotal).

the following indirect indicators of the effort ca.48. using a cycloergometer.0001 out for 6 minutes. Results • The maximum O2 consumption (VO2max) expressed in ml and calculated by the tailored The mean age of the study group was 24.31 < 0. 24. • The cardiovascular economy in effort.03 *The parameters are expressed as Mean ± SD.31 the rotation of 40-80/min and the intensity of HGB (g/dl) 15.50 < 0.0001 Crp (mg%) 25. ing Graph Pad Prism 5 software. highlight the qualitative differences between the pacity were calculated: variables: the mean. The Astrand-Ryhming ± 0. and consisted of an indirect determina- tion of the aerobic effort capacity using the As.05 was considered sta- = VO2max/kg.14 < 0. the • The maximum aerobic power.03 < 0.11 ± 0. The following The results for the oxidative and antioxida- statistical indicators were calculated so as to tive balance parameters are shown in Table 2. Post.01 0.26 kg.62 ± 0. rate. workload + 0. On the basis of the measured * The parameters are expressed as Mean ± SD.0001 HD (%) 45.37 ± 0.92 ± 57.98 ± 3.769 x HRss . 2016 The exploration of the effort capacity was Table 1. The oxidative and antioxidative balance parameters Oxidative Pre. Nr.13 6 6.22 ± 2. . Table 2. Post.0001 (nmol/ml) 0.04 ± < 0.330 Revista Română de Medicină de Laborator Vol.09 test consisted of a submaximal effort carried HCT (%) 44. RBC (10 /µl) 5. with ± 1.05.0001 trand-Ryhming method. p value from Brașov.01 < 0.00212 x 1. which is located at an altitude of training training 592 m.299) / (0. ing for the hematological parameter.0001 175 W/kg.47 ± 7.11 ± formula for men [9] VO2max = (0. heart rate during the last 10 seconds of pedaling. Normality of the data was assessed using the pressed in ml/min and calculated using the Shapiro-Wilk test and the significance level was formula CVEE = VO2max/Maximum heart set at α=0. ex.0001 UA (mg/dL) 3. Pre. the standard deviation.04 0.45 years and the mean weight 69. p value balance pa.98 ± < training tive balance training training rameters* parameters* LPx 1. Septembrie. tistically significant.08 0. The red blood cell parameters assessed at the County Medical Center MAI Parameter* Pre.0001 (nmol/ml) 0.33 ± 0.69 ± 0.06 total MDA 2.32 ± 1. tween pre-training testing and post-training test- The statistical analysis was performed us.02 < 0. expressed in minimum and maximum values. p value Antioxida.20 ± 1.0001 (nmol/ml) 0.73 ± 0.01 < 0. 3. A p value < 0.25 test procedure [8].13 0. Post.13 0.61 ± 1.06 17.80 51. maintained constant during the whole ± 0. the Student’s ml/kg and calculated using the formula MAP t-test [10].5) Table 1 indicates a significant difference be- x 100.95 ± 31.12 free MDA 1.

45 5945 ± 46. Septembrie. 0. stimulates the production and Some authors [8. we found that the means were superior in the Acclimatization and training protocols like liv. Comparing the results of our study with the Discussions ones obtained by Martoma on alpine skiers. alkaline reserve. associated with an 18 g/dl and the HCT increases from 40-45% to increase in VO2max. 0. from the sea or LH-TL model has advantages over SL train.48 g/dl for HGB and poxic exposure (IHE) protocol provided natural 3. Nr.53 ± 1. during the first 24-48 Chapman et al. RBC. liv. the functional state presents no relevant tion in the LH-TL according to the new solution: changes.13 ± 0. etc. .training high protocol (Hi-Hi). 11] consider that there is the discharge of erythropoietin (EPO). Ev. which induces a low yield for the while high-intensity interval training should be next seven to ten days.41 ± 0. provement of several hematological parameters tathione. 24. 14. from the third athletes should stay and perform low-intensity day after exposure to altitude. 2016 331 Table 3.97 < 0. the Hi-Hi-Lo protocol [4. it was found that after a train. 12]. After this acclimatization performed at 1250 m.0001 VO2max (ml) 4190 ± 50. In this respect. 15]. The parameters showing the aerobic effort capacity Parameter* Pre-training Post-training p value MAP (ml/kg) 63. glu. mean differences among the tests between the ing high-training low protocol (Hi-Lo). 3.48 < 0. the acclimatization training at an altitude of 2500 m for four weeks. and norepineph- power output and economy [5]. ing period of 21-28 days. favor of the biathlon group [11. rine. respectively. in which aer. The parameters showing the aerobic effort Hematological parameters capacity are presented in Table 3.16x106/µl for RBC. in significant improvement of running time after with a maximization peak between days 14 to 18.57 57. Revista Română de Medicină de Laborator Vol. other studies found a signif- biochemical parameters reach very good values icant increase in VO2max accompanied by im- (HGB.). to levels of over ing to improve performance.50% for HCT. Renal hypoxia. blood cells from the progenitor cells [13]. but after this short period. Thus. living biathlon group and alpine skiing group were: low-training high (Lo-Hi). and intermittent hy. VO2max. Thus. more than 55%.63 < 0. and artificial methods that have been developed Studies have shown that the red cell mass to improve aerobic and anaerobic capacity. slowly increases along with prolonged hypox- idence in the literature suggests that the Hi-Lo emia.0001 * The parameters are expressed as Mean± SD.0001 CVEE (ml/min) 33. level value around 14-15 g/dl. HGB concentration increases. This method was called phase the positive phase follows. Hi-Hi-Lo (live high-base train high-interval train obic efficiency can increase by up to 15% and low). phase installs.00 81. Sport yield is good in the range of (hemoglobin mass and hematocrit) that resulted 14 to 21 days after exposure to medium altitude. [14] proposed one modifica- hours. hematological parameters. EPO is a an “acclimatization memory” that is expressed growth factor that stimulates the production of through an accentuated and gradual attenuation proeritroblasts in the bone marrow and deter- of the adaptive changes to the training sessions mines the acceleration of the development of red at altitude. the ing high.91 ± 1.

durance athletes’ training. be explained by the maintenance of a high arte. and in terms of antiox- the maximum effort capacity or the VO2max idative balance parameters. This determines the de- antioxidative balance crease of the glucose transport in muscles. (1800-1900 m) showed that supplementation cient ‘altitude’ our results highlight that LHTH in O2 on Hi-Lo models determines a significant is the most time efficient modality for athletes increase of the physical performance and a de- and coaches to consider. Acclimatization to high pect a ~3% increase in HGB mass after only 14 altitude does not only increase the release of the days of LHTH training at 1800 m. glutathione) and urine (urinary malondialdehyde crease in HGB mass in the third week [23]. and 8-hidroxi-deoxi-guanine) [24]. dl for UA. in on altitude to the pre-acclimatization values. showed an in. with direct effect on the motor capacity. The aerobic effort decreases the sensitivity The determinations of the oxidative and of the tissue to insulin. respectively. This lowers O2 from the periphery but also the O2 uptake by the threshold altitude considered sufficient to in. 18 hours a day and train rameters recorded significant increases in condi- at 1000 and 1800 m for 24 days. Septembrie. terms of the oxidative balance parameters. because it provides a crease in the oxidative stress indicators. the tissues [11. 2500. rial desaturation that is produced by the limited Hypoxia is a promoter of the oxidative en- diffusion [21]. 12]. elite athletes. it should be noted that the biathlon group re- hypoxic stimulus is removed [13. which pared to the studies performed on alpine skiers by may help to increase the performance of endur. tions of altitude training in biathlon athletes com- creased HGB mass and red cell volume. 24. the O2 from the blood. 0. Nr. reflect- 24-hour per day exposure to hypoxia [22]. 12]. A ed in the blood (lipid hydroperoxide and reduced meta-analysis highlighted an additional ~2% in. at a moderate altitude tion seems to depend on the total dose of suffi. This can thus contributing to a better adaptation to altitude.04 nmol/ml for LPx. and on athletes in real conditions [11. determin- The results of another study have illustrated ing the increase of the tissue ability to extract that it is possible for endurance athletes to ex.05 nmol/ml for MDAtotal. both initially and finally. Analyzing the results obtained ance athletes [16]. 1.01 nmol/ml for MDAfree. 12]. The oxidative and antioxidative balance pa- who live at 2500 m. Although these differences are not very crease in HGB and HCT starting as soon as the high. The effect of the O2 fractions crease the red blood cell production by ~200m. ance changes in the case of medium altitude .12 mg/dl for Crp and 0. 0. 0. the differences between the mean values. 3. mountain guides. in- creasing the reconversion into free fatty acids Studies on the oxidative and antioxidative bal- and saving the muscle glycogen [25. 1000. durance athletes. zymes expression in the mitochondria. The acute HGB increase does not affect 0. 2016 The Hi-Lo effect on the quantity of HGB and training were simulated both on laboratory rats on the volume of RBC studied on resistance/ en. the differences were: during the effort in acute hypoxia [17].06% for HD. and 4500 m. Martoma [11. but especial- is a decrease of the VO2max at the altitude of ly in the case of antioxidative balance parameters. 26].332 Revista Română de Medicină de Laborator Vol. on For the athletes trained for endurance there all the oxidative balance parameters. in our study with those obtained on the alpine ski- The HGB decrease in subjects acclimatized ers. were: does not affect VO2max during the effort in hy. 18-20]. poxia. corded higher values. inspired upon the oxygenation during the en- Given that the accelerated red blood produc.11 mg/ Several studies show a relatively linear de.

29-31]. for CVEE were titions are performed at submaximal intensities. 28]. during the LH-TL The comparative analysis of our 2014 re- protocol. 11. Moreover. [11. minimum period of three weeks at altitude will Numerous studies showed the effects of alti. in 2014 observed a signifi- The determination of the aerobic effort ca. With caused by the decrease of O2 availability under the return to low altitude. such as the potential tion time increases according to the increase in factors influencing the aerobic capacity and the altitude and the physiological changes are more endurance performance. by the specificity of the practiced sport and the . re- VO2max in spite of the significantly higher val. since most of the biathlon compe- 8.40 ml/kg at the initial testing and in normoxia. Anaerobic trials are not affected by the help athletes to improve their performance ca- moderate altitude while aerobic trials are [3. allows an optimal acclimatization re- corded results (the same group of biathletes) sponse in sea level performance [32].40 ml/min at the initial testing and 8. Revista Română de Medicină de Laborator Vol. highlights statistically significant lower values The positive changes observed in our partic- than those obtained in the current research. cause physiological changes in the body in order tude on the physical performance of the athletes. The supe- tation through the correlation of some other mor- rior results of the biathlon group are influenced pho-functional and motor parameters. 27. Additionally. cant improvement of the sport performance in pacity parameters groups living at two medium altitudes (2085 and 2454 m) for four weeks. between 2000 and 2500 m. not improve the aerobic capacity estimated by taking place in a few days or on a long-term. The adapta- variables were evaluated. 12].40 ml/kg at the final testing. 3. pacity. ditions affects biathletes’ aerobic capacity and Studies on effort physiology prove that effort energy cost in normoxia. of participants is a limitation of the present in- Comparing the alpine skiers’ groups of Mar- vestigation. in order to allow acclimatization to high study was that the Hi-Hi-Lo 3-week protocol did altitude. 1. endurance sports. ipants led to the conclusion that Hi-Hi-Lo train- the differences between the two researches for ing method could effectively improve endurance MAP were 1. dium altitude. these adjustments will hypoxia. 12] with our biathlon group. EPO levels were cant increases in the post-training testing at me- significantly higher in all groups at 24 and 48 h. toma et al. These results suggest that. but not in groups living The aerobic capacity parameters showed signifi- at 1780 and 2800 m. 24. 11. Adapting can have immediate effects.the altitude tor potential of the athletes. compared with the initial testing. quiring weeks or even months [8. A ues of the hematological parameters [4]. Thus. Nr. The main finding of the intense. Chapman et al. the 1800-2400 m altitude area is the ideal place for erythropoietin and the selected hematological performing sports training stages. 2016 333 Another study investigated if the Hi-Hi-Lo athlete’s selection and training are customized to procedure performed in hypo-baric hypoxia con. but returned to basic levels after 72 h in the 1780 and contributing to the increase of the effort mo- m group. Septembrie. to adapt to a better use of the received O2. ed by the ability to use O2 [11. because the skiers’ performance is limit- 12. the Our future research will focus on the indi- mean of the recorded results of the biathletes is vidual differences in terms of the altitude adap- above the mean of the skiers group.90 ml/min The fact that the study was conducted with at the final testing and for VO2max of 72 ml at a convenience sample of a fairly small number the initial testing and 200 ml at the final testing.

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