Program Protocol for a Population That Works in Conditions of Hypobaric Hypoxia

This study is an investigation that seeks to evaluate the effect of a physical exercise and nutritional protocol on the determinants of health (nutritional status and body composition, metabolic-inflammatory parameters, glucose profile, gut microbiome, physical condition, autonomic nervous system, and sleep patterns) of workers who perform their workday at an astronomical observatory under hypobaric hypoxia conditions between 2,950 and 5,050 meters above sea level. The objective of this intervention is to standardize both physical and nutritional recommendations for this type of worker under intermittent hypobaric hypoxia conditions.

The procedure will begin by incorporating personnel who meet the inclusion criteria: (being an employee contracted by the ALMA Observatory with more than one year of seniority, working in AOS or OSF sectors of the observatory, and not suffering from non-communicable diseases such as type 2 diabetes or hypertension). On the other hand, the exclusion criteria for this research are: people who use pacemakers, those who are temporarily stationed at the facilities, and pregnant women. Once the "n" of 11 subjects for each research group has been obtained, all participants will undergo a nutritional and body composition assessment by a registered dietitian. This assessment will utilize a multifrequency bioimpedance analysis (BIA) device, model INBODY 270S. Participants will be asked to come to the designated laboratory after fasting for 4 hours and removing as much clothing as possible, including shoes, socks, and any metal jewelry. The analysis will consist of weighing the participant in kilograms and measuring their height (SECA 700 scale). Subsequently, waist and hip circumferences will be measured using a SECA 201 ergonomic measuring tape. To complete the nutritional assessment, participants will be asked to step onto the BIA device barefoot, where they will place the hand electrodes and follow the evaluation instructions provided by the device. Subsequently, a Polar H10 elastic heart rate monitor will be fitted to the chest area and connected to the Polar Beat app for continuous heart rate monitoring. Along with this heart rate monitor, each participant will also have an Actigraph WG3XBT elastic heart rate monitor fitted to their hip area to measure the intensity of their movements. To determine the physical condition and estimate the VO2max of the subjects under study, all will undergo a Modified Queens College box test, in a 20 cm high box for a period of 3 minutes. Finally, a Freestyle Libre 2 Plus continuous glucose monitoring patch will be fitted to the left triceps. These individually fitted devices will continue to collect data for 14 consecutive days. Once the entire installation phase is complete, each participant will be given two sealed, sterilized plastic containers. In these containers, they must provide the research team with two small fecal samples (one on the day of arrival at the observatory and another on the last day of their stay). These samples will be processed in the laboratory set up for this purpose at the observatory using a QIAamp PowerFecal Pro DNA Kit. A DNA sample will be extracted following the kit's instructions, then frozen and transported to the University facilities. There, the samples will be processed using a DNA sequencing system. They will be stored in the buffer included with the aforementioned extraction kit and subsequently frozen in a special -20°C refrigerator at the GEMA laboratory for later analysis. The samples will then be analyzed using an Invitrogen Qubit fluorometer to quantify the concentration of extracted DNA. Subsequently, the samples will be analyzed using capillary electrophoresis on a Bioptic Fragment Analyzer™ (QSep 1). Following this, the libraries will be prepared according to the sequencer to be used, and sequencing will then begin on an Illumina MiSeq and/or Nanopore MinION platform (depending on the number of DNA samples extracted and the amount of data to be sequenced).

In addition, a trained paramedic will collect 5 ml of venous blood, which will be placed in polypropylene tubes with anticoagulant and centrifuged at 3000 rpm. The plasma and blood will then be sent refrigerated to the CEIMA laboratory at Arturo Prat University for analysis and quantification of inflammatory factors using the SOD Colorimeter Activity Kit and the GSH-PX Activity Kit, following the respective manufacturers' instructions.

Once the initial analyses are complete, the study subjects will be divided into two groups:

G1 (Intervention group with resistance training and nutritional supplementation) This group will undergo a resistance training program guided by a physical activity professional for three days during their work shift, along with a daily intake of a probiotic and prebiotic supplement for seven days. To this end, the maximum number of repetitions (1RM) that the individual can perform for each of the four muscle groups will be calculated at the beginning. Therefore, 3 sets of 10 repetitions each will be performed for the following muscle groups: 1. biceps, 2. quadriceps, 3. pectorals, and 4. gastrocnemius, at 50% of the previously calculated maximum repetition.

In addition, this group's diet will be supplemented with the administration of two "Karún Life" brand probiotic tablets for the seven days of the work shift.

G2 (Overload Physical Exercise Intervention Group) This group will undergo the same overload training program as the previous group under the same conditions mentioned above.