Study protocol for evaluating the current status and needs assessment of health-related characteristics among students at Albert-Ludwigs-University Freiburg.

Today, university students face study conditions that increase the risk of sedentary behavior, unhealthy eating, and the likelihood of stress, anxiety, and depression. Although the situation has likely worsened in recent months due to the COVID-19 pandemic, even in the pre-Corona era, several investigations have demonstrated that the students' health behaviors could increase the incidence of metabolic risk factors and non-communicable diseases, however, further and detailed information is needed to develop tailored counter-strategies. Therefore, in the present survey, the current health status of students at the Albert-Ludwigs-Universität Freiburg (ALU) will be recorded by various questionnaires. In addition, barriers that could potentially influence their health behavior will be identified, and information on the use and awareness of existing health services will be obtained in order to derive concrete needs for further health promotion activities. The study is designed as a monocentric and prospective study at the ALU; the survey of students' situation and needs assessments will be conducted in the areas of nutrition, exercise, relaxation and stress reduction, self-management, psychosocial health and socio-demographic correlations via an anonymous and self-administered online questionnaire. Subsequently in two-year intervals, the survey will be repeated. Data will be collected over a period of 4 years. One goal of this survey is to gain more insight into the health situation of university students; another goal is to use the knowledge gained to integrate holistic health measures into the university landscape in a structured and sustainable manner. These health measures will be implemented by using the newly developed student health care management at the ALU (www.sgm.uni-freiburg.de). Every two years, after evaluation of the surveys, the effect of the health measures can be checked and adjusted. Trial registration: ETK: 20-1082; DRKS-ID: DRKS00024088.

Effects of specific collagen peptide supplementation combined with resistance training on Achilles tendon properties.

The purpose of this study was to investigate the effect of specific collagen peptides (SCP) combined with resistance training (RT) on changes in tendinous and muscular properties. In a randomized, placebo-controlled study, 40 healthy male volunteers (age: 26.3 ± 4.0 years) completed a 14 weeks high-load resistance training program. One group received a daily dosage of 5g SCP while the other group received 5g of a placebo (PLA) supplement. Changes in Achilles tendon cross-sectional area (CSA), tendon stiffness, muscular strength, and thickness of the plantar flexors were measured. The SCP supplementation led to a significantly (p = 0.002) greater increase in tendon CSA (+11.0%) compared with the PLA group (+4.7%). Moreover, the statistical analysis revealed a significantly (p = 0.014) greater increase in muscle thickness in the SCP group (+7.3%) compared with the PLA group (+2.7%). Finally, tendon stiffness and muscle strength increased in both groups, with no statistical difference between the groups. In conclusion, the current study shows that the supplementation of specific collagen peptides combined with RT is associated with a greater hypertrophy in tendinous and muscular structures than RT alone in young physically active men. These effects might play a role in reducing tendon stress (i.e., deposition of collagen in load-bearing structures) during daily activities.

Specific Bioactive Collagen Peptides in Osteopenia and Osteoporosis: Long-Term Observation in Postmenopausal Women.

BACKGROUND: The effects of specific collagen peptides on bone mineral density (BMD) in subjects with osteoporosis or osteopenia have already been investigated in 131 postmenopausal women in a randomized controlled trial. The purpose of this follow-up observation was to determine the longer-term effects of the same specific bioactive collagen peptides after a total intervention time of 4 years. METHODS: In this open-label follow-up observation, 31 postmenopausal women with reduced BMD (initial T-score lower than-1 of either the femoral neck or the lumbar spine) completed the follow-up. BMD was measured via dual energy X-ray absorptiometry. Absolute changes in BMD and T-scores in the spine and femoral neck were assessed. The number of fractures was also recorded. All participants received specific bioactive collagen peptides. RESULTS: Supplementation with bioactive collagen peptides during follow-up led to a clinically relevant increase in BMD in the spine. These findings were consistent with the results for the femoral neck. CONCLUSIONS: Long-term supplementation with specific bioactive collagen peptides appears to be effective in counteracting losses in BMD. Moreover, significant increases in BMD could contribute to improved bone stability.

The Influence of Specific Bioactive Collagen Peptides on Body Composition and Muscle Strength in Middle-Aged, Untrained Men: A Randomized Controlled Trial.

It has been shown that specific collagen peptides combined with resistance training (RT) improves body composition and muscle strength in elderly sarcopenic men. The main purpose of this RCT study was to investigate the efficacy of the identical specific collagen peptides combined with RT on body composition and muscle strength in middle-aged, untrained men. Furthermore, in the exploratory part of the study, these results were compared with another group that had received whey protein in addition to the RT. Ninety-seven men completed this study and participated in a 12-week RT program. They ingested 15 g of specific collagen peptides (n = 30; CP-G), placebo (n = 31; P-G), or whey protein (n = 36; WP-G) daily. Changes in fat free mass and fat mass were determined by dual-energy X-ray absorptiometry (DXA), and isometric leg strength was measured. All participants had significantly (p < 0.01) improved levels in fat free mass (ΔCP-G = 3.42 ± 2.54 kg; ΔP-G = 1.83 ± 2.09 kg; ΔWP-G = 2.27 ± 2.56 kg), fat mass (ΔCP-G = -5.28 ± 3.19 kg; ΔP-G = -3.39 ± 3.13 kg; ΔWP-G = -4.08 ± 2.80 kg) and leg strength (ΔCP-G = 163 ± 189 N; ΔP-G = 100 ± 154 N; ΔWP-G = 120 ± 233 N). The main analysis revealed a statistically significantly higher increase in fat free mass (p = 0.010) and decrease in fat mass (p = 0.023) in the CP-G compared with the P-G. The exploratory analysis showed no statistically significant differences between WP-G and CP-G or P-G, regarding changes of fat free mass and fat mass. In conclusion, specific collagen peptide supplementation combined with RT was associated with a significantly greater increase in fat free mass and a decrease in fat mass compared with placebo. RT combined with whey protein also had a positive impact on body composition, but the respective effects were more pronounced following the specific collagen peptide administration.

The Influence of Specific Bioactive Collagen Peptides on Knee Joint Discomfort in Young Physically Active Adults: A Randomized Controlled Trial.

First evidence indicates that the supplementation of specific collagen peptides is associated with a significant reduction in activity-related joint pain in young adults. The purpose of the current investigation was to confirm the efficacy of the same collagen peptides in a comparable study population. In total, 180 active men and women aged between 18 and 30 years with exercise-related knee pain but no diagnosed joint disease completed the trial over a period of 12 weeks. Participants were randomly assigned to the group receiving 5 g of specific collagen peptides (CP-G) or to the placebo group (P-G). For the primary outcome, changes in pain during or after exercise from pre- to post-intervention were assessed by the participants using the Visual Analog Scale (VAS). These changes were additionally evaluated by the examining physician by means of anamnesis and physical examination of the affected knee joint. As secondary outcomes, pain under resting conditions and after 20 squats were compared between the study groups. In addition, the mobility of the knee joint and the use of alternative therapies (e.g., ointments or physiotherapy) were recorded. The supplementation of specific collagen peptides derived from type I collagen with a mean molecular weight of 3 kDa led to a significantly (p = 0.024) higher reduction of exercise-induced knee pain (-21.9 ± 18.3 mm) compared with the placebo group (-15.6 ± 18.5 mm). These findings were consistent with the physician's evaluation (-23.0 ± 19.2 mm vs. -14.6 ± 17.9 mm, p = 0.003). The decrease in pain under resting conditions and after squats did not significantly differ between the groups, as only a small number of participants suffered from pain under these conditions. Due to the clinically unremarkable baseline values, the mobility of the knee joint did not change significantly after the intervention. In conclusion, the current investigation confirmed that the oral intake of bioactive collagen peptides used in the current investigation led to a statistically significant reduction of activity-related joint pain in young active adults suffering from knee joint discomfort.

Effects of Blood Flow Restriction Training with Protein Supplementation on Muscle Mass And Strength in Older Men.

Blood flow restriction (BFR) training has been shown to induce favorable changes in muscle mass and strength with a considerably low training load (20 - 30% 1RM). However, it has never been evaluated if an additional post-exercise protein supplementation enhances the effects of this training regimen. Thirty healthy older men (60.1 ± 7.6 years) were enrolled in the 8-week intervention and randomly allocated to one of the following groups: low-load BFR training with protein (collagen hydrolysate) supplementation (BFR-CH), low-load BFR training with placebo (BFR-PLA), or a control group without training, but with protein supplementation (CON). Muscle cross-sectional area (CSA), muscle strength, circulating reactive oxygen species and IGF-1 were measured before and after the intervention. Muscle CSA increased in both BFR-CH and BFR-PLA groups by 6.7 ± 3.2 % (p < 0.001) and 5.7 ± 2.7 % (p < 0.001) respectively. No significant changes were observed in the CON group (1.1 ± 1.7 %, p = 0.124). Evaluation of isometric strength (p = 0.247), insulin-like growth factor 1 (p = 0.705) and the production of reactive oxygen species (pt1 = 0.229; pt2 = 0.741) revealed no significant interaction effect but a significant long-term time effect (p < 0.001). Our results demonstrate that BFR training is an effective alternative for increasing muscle CSA in older men. Although there was a trend towards greater muscle mass adaptations in the BFR-CH group, these findings showed no statistical significance. Further research with larger sample sizes is needed to confirm these results.

Specific Collagen Peptides in Combination with Resistance Training Improve Body Composition and Regional Muscle Strength in Premenopausal Women: A Randomized Controlled Trial.

The aim was to investigate the effects of resistance exercise combined with supplementation of specific collagen peptides (SCP) on body composition and muscle strength in premenopausal women. In a double-blind, placebo-controlled, randomized trial 77 premenopausal women completed a 12-week resistance training (3 day/week) and ingested 15 g of SCP or placebo on a daily basis. Changes in body composition were determined by bioelectrical impedance analysis (BIA) and muscular strength by isometric strength testing. The treatment group (TG) significantly increased (p < 0.001) their percentage of fat-free mass. Although the control group (CG) also showed a significant (p < 0.01) gain in fat-free mass from pre- to post-training, the increase in the TG was significantly higher in an RMANOVA analysis (p < 0.05). Regarding the change in percentage body fat, a significant decline was observed in both TG (p < 0.001) and CG (p < 0.01), with a significantly higher reduction in the TG (p < 0.05). Subjects receiving 15 g of collagen peptides daily also showed a significantly higher gain in hand-grip strength compared to those performing resistance training only (p < 0.05). In both groups, the gain in leg strength (TG = p < 0.001; CG = p < 0.01) was significant after 12 weeks with a more pronounced effect in the treatment group. In conclusion, resistance training in combination with supplementation of SCP induced a significantly higher increase in fat-free mass and hand-grip strength than resistance training and placebo supplementation. In addition, there was a significantly higher loss in fat mass and a more pronounced increase in leg strength in the treatment group compared to the control group.

The influence of an anti-inflammatory diet on gingivitis. A randomized controlled trial.

AIM: Aim of this study was to investigate the influence of an anti-inflammatory diet on different parameters in patients with gingivitis. MATERIALS AND METHODS: Thirty patients were randomly allocated to an experimental and a control group stratified by their plaque values. The experimental group had to change to a diet low in processed carbohydrates and animal proteins, and rich in omega-3 fatty acids, vitamin C, vitamin D, antioxidants, plant nitrates and fibres for 4 weeks. The control group did not change their diet. Both groups suspended interdental cleaning. Periodontal parameters were assessed by a blinded dentist. Serological and subgingival plaque samples were taken at baseline and end. RESULTS: While there were no differences regarding the plaque values, the experimental group showed a significant reduction in gingival bleeding (GI Baseline: 1.04 ± 0.21, GI End: 0.61 ± 0.29, p < 0.05), a significant increase in Vitamin D values and a significant weight loss. There were no inter-group differences regarding the inflammatory serological parameters, the serological omega fatty acids, nor the subgingival microbiome composition. CONCLUSION: The evaluated diet could significantly reduce gingivitis in a clinically relevant range, while serological inflammatory parameters and the subgingival microbiome seem to be unaffected in this study duration. (German Clinical Trials Register; DRKS00009888).

Improvement of Functional Ankle Properties Following Supplementation with Specific Collagen Peptides in Athletes with Chronic Ankle Instability.

Following an initial ankle sprain it is not unlikely that chronic ankle instability (CAI) will develop. CAI is associated with impaired perceived functional and mechanical properties of the ligaments. Nutritional supplementation with collagen peptides has been shown to improve the functional and mechanical properties of the connective tissue. The purpose of this study was to investigate the effectiveness of specific collagen peptide supplementation (SCP) to improve ankle stability in athletes with CAI. 50 male and female athletes with CAI completed a randomized, double-blinded and placebo-controlled study with a daily oral administration of either 5 g SCP or 5 g placebo (Maltodextrin) over a period of six months. Both, the Cumberland Ankle Instability Tool (CAIT) and the German version of the Foot and Ankle Ability Measure (FAAM-G) were used to measure the subjective perceived function of the ankle. Additionally, the mechanical stability was determined by measuring the ankle stiffness by an ankle arthrometer. Finally, a three-month follow-up was performed. ANOVA analysis indicated that the subjective ankle stability was improved in both the CAIT (p < 0.001) and the FAAM-G (p < 0.001) following SCP supplementation compared with placebo. No significant changes between the groups were detected in the results of the ankle arthrometer. After six month the subjective report of the ankle stability function significantly improved and the three month follow-up revealed a significant decline in the number of ankle joint injuries (p < 0.05). These data support the concept that specific collagen peptide supplementation in athletes with chronic ankle instability results in significant improvements in subjective perceived ankle stability. The reduction in the re-injury rate of ankle sprains in the follow-up period suggests that these findings have clinical relevance.

Specific Collagen Peptides Improve Bone Mineral Density and Bone Markers in Postmenopausal Women-A Randomized Controlled Study.

Introduction: Investigations in rodents as well as in vitro experiments have suggested an anabolic influence of specific collagen peptides (SCP) on bone formation and bone mineral density (BMD). The goal of the study was to investigate the effect of 12-month daily oral administration of 5 g SCP vs. placebo (CG: control group) on BMD in postmenopausal women with primary, age-related reduction in BMD. Methods: 131 women were enrolled in this randomized, placebo-controlled double-blinded investigation. The primary endpoint was the change in BMD of the femoral neck and the spine after 12 months. In addition, plasma levels of bone markers-amino-terminal propeptide of type I collagen (P1NP) and C-telopeptide of type I collagen (CTX 1)-were analysed. Results: A total of 102 women completed the study, but all subjects were included in the intention-to-treat (ITT) analysis (age 64.3 ± 7.2 years; Body Mass Index, BMI 23.6 ± 3.6 kg/m²; T-score spine -2.4 ± 0.6; T-score femoral neck -1.4 ± 0.5). In the SCP group (n = 66), BMD of the spine and of the femoral neck increased significantly compared to the control group (n = 65) (T-score spine: SCP +0.1 ± 0.26; CG -0.03 ± 0.18; ANCOVA p = 0.030; T-score femoral neck: SCP +0.09 ± 0.24; CG -0.01 ± 0.19; ANCOVA p = 0.003). P1NP increased significantly in the SCP group (p = 0.007), whereas CTX 1 increased significantly in the control group (p = 0.011). Conclusions: These data demonstrate that the intake of SCP increased BMD in postmenopausal women with primary, age-related reduction of BMD. In addition, SCP supplementation was associated with a favorable shift in bone markers, indicating increased bone formation and reduced bone degradation.

Oxidative stress or redox signalling - new insights into the effects of a proprietary multifunctional botanical dietary supplement.

Recent interest has focused on maintenance of healthy levels of redox signalling and the related oxidants; these parameters are crucial for providing us with concrete nutritional targets that may help us to better understand and maintain "optimal health". Following the above hypothesis, we performed a pilot double-blind, crossover, placebo-controlled, single dose study to measure the dose-dependent effects of a proprietary plant-based dietary supplement labelled here as S7 (SPECTRA7), related to how it affected the cellular metabolic index (CMI) in healthy human participants (n = 8). We demonstrated using the electron spin resonance/electron paramagnetic resonance spectrometer NOXYSCAN that the administration S7 resulted in statistically significant, long-term, dose-dependent inhibition of mitochondrial and cellular reactive oxygen species generation by as much as 9.2 or 17.7% as well as 12.0 or 14.8% inhibition in extracellular nicotinamide-dinucleotide-phosphate oxidase system-dependent generation of O2•-, and 9.5 or 44.5% inhibition of extracellular H2O2 formation. This was reflected with dose-dependent 13.4 or 17.6% inhibition of tumour necrosis factor alpha induced cellular inflammatory resistance and also 1.7 or 2.3-times increases of bioavailable NO concentration. In this pilot study, we demonstrated the ability of a natural supplement to affect cellular redox signalling, which is considered by many researchers as oxidative stress. The design and activity of this proprietary plant-based material, in combination with the newly developed "CMI" test, demonstrates the potential of using dietary supplements to modulate redox signalling. This opens the door to future research into the use of S7 for modulation of inflammatory markers, for sports endurance or recovery applications.

Improvement of activity-related knee joint discomfort following supplementation of specific collagen peptides.

The aim of the study was to evaluate the use of specific collagen peptides in reducing pain in athletes with functional knee problems during sport. Athletic subjects (n = 139) with functional knee pain ingested 5 g of bioactive collagen peptides (BCP) or a placebo per day for 12 weeks. The primary outcome of the study was a change in pain intensity during activity, which was evaluated by the participants and the attending physicians using a visual analogue scale (VAS). As secondary endpoints, pain intensity under resting conditions, the range of motion of the knee joint, and the use of additional therapeutic options were assessed. The results revealed a statistically significant improvement in activity-related pain intensity in the verum group compared with placebo. (ΔVASBCP = 19.5 ± 2.4; ΔVASPlacebo = 13.9 ± 2.1; p = 0.046). The results were confirmed by the physician's assessment. (ΔVASBCP = 16.7 ± 1.8; ΔVASPlacebo = 12.2 ± 1.8; p = 0.021). Pain under resting conditions was also improved, but no significance compared with placebo was detected (ΔVASBCP = 10.2 ± 18.4; ΔVASPlacebo = 7.4 ± 15.2; p = 0.209). Due to the high joint mobility at baseline, no significant changes of this parameter could be detected. The use of additional treatment options was significantly reduced after BCP intake. The study demonstrated that the supplementation of specific collagen peptides in young adults with functional knee problems led to a statistically significant improvement of activity-related joint pain.

Collagen peptide supplementation in combination with resistance training improves body composition and increases muscle strength in elderly sarcopenic men: a randomised controlled trial.

Protein supplementation in combination with resistance training may increase muscle mass and muscle strength in elderly subjects. The objective of this study was to assess the influence of post-exercise protein supplementation with collagen peptides v. placebo on muscle mass and muscle function following resistance training in elderly subjects with sarcopenia. A total of fifty-three male subjects (72·2 (sd 4·68) years) with sarcopenia (class I or II) completed this randomised double-blind placebo-controlled study. All the participants underwent a 12-week guided resistance training programme (three sessions per week) and were supplemented with either collagen peptides (treatment group (TG)) (15 g/d) or silica as placebo (placebo group (PG)). Fat-free mass (FFM), fat mass (FM) and bone mass (BM) were measured before and after the intervention using dual-energy X-ray absorptiometry. Isokinetic quadriceps strength (IQS) of the right leg was determined and sensory motor control (SMC) was investigated by a standardised one-leg stabilisation test. Following the training programme, all the subjects showed significantly higher (P<0·01) levels for FFM, BM, IQS and SMC with significantly lower (P<0·01) levels for FM. The effect was significantly more pronounced in subjects receiving collagen peptides: FFM (TG +4·2 (sd 2·31) kg/PG +2·9 (sd 1·84) kg; P<0·05); IQS (TG +16·5 (sd 12·9) Nm/PG +7·3 (sd 13·2) Nm; P<0·05); and FM (TG -5·4 (sd 3·17) kg/PG -3·5 (sd 2·16) kg; P<0·05). Our data demonstrate that compared with placebo, collagen peptide supplementation in combination with resistance training further improved body composition by increasing FFM, muscle strength and the loss in FM.

A soy-based supplement alters energy metabolism but not the exercise-induced stress response.

OBJECTIVE: To determine the changes in endurance capacity as well as in metabolic, hormonal and inflammatory markers induced by endurance training combined with a soy,protein based supplement. DESIGN: Randomized controlled study consisting of moderate endurance training without (GO) or with (G1) a soy protein based supplement. SUBJECTS: Two groups of 15 subjects (10 males and 5 females in each group): healthy sports students aged 23.6 +/- 1.9 years. MEASUREMENTS: Body composition (body mass (BM), body density (BD) by air displacement) and physical fitness (determined by treadmill ergometry) were measured at baseline and after 6 weeks of the intervention; changes in circulating metabolic and hormonal parameters (glucose, lactate, urea, uric acid, ammonia, cortisol, insulin, IGF-1), and exercise-induced stress and inflammatory markers (CK, LDH, myoglobin, hs-CRP, IL-6, IL-10, blood cell counts) were determined after the intervention period in afield test (11.5 km running on hilly ground). RESULTS: 30 participants completed the 6-week study; 28 students were able to perform the field test. No significant changes in BM and BD were noted after intervention with only slight increases in running performance and maximum aerobic capacity in the total group (2%, p=0.016). Subjects in the G1 group showed significant improvements in running velocity and lower lactate values following the intervention (-12%, p=0,003). In addition, the G1 group showed significantly lower differences in the exercise-induced increase of metabolic parameters (triglycerides, uric acid) and insulin in the post-exercise recovery period. CONCLUSIONS: Our data suggest that moderate endurance training in combination with a soy-based protein supplement improves aerobic energy supply and metabolic function in healthy sports students, even without changes in body composition and without changes in the exercise-induced stress and inflammatory reaction.

Soy protein based supplementation supports metabolic effects of resistance training in previously untrained middle aged males.

OBJECTIVE: To determine changes in body composition, physical performance, metabolic and hormonal parameters induced by lifestyle counselling, resistance training and resistance training with soy protein based supplemention in middle aged males. DESIGN: Randomised controlled study consisting of resistance training without (RT-G) or with (RTS-G) a soy protein based supplement and a control group with lifestyle education only (LE-G). SUBJECTS: Forty healthy middle aged men (50-65 years, BMI 25-29.9 kg/m2). MEASUREMENTS: Changes in body weight (BW) and waist circumference (WC) were measured and body composition (BC), fat mass (FM), lean body mass (LBM) were measured by skin fold anthropometry at baseline and after 12 weeks of intervention. In addition, changes in physical fitness, metabolic and hormonal parameters (lipids, glucose, fructosamines, insulin, insulin-like growth factor-1, Leptin, human growth hormone, dehydroepiandrosterone, testosterone, hs-CRP, Il-6) were evaluated. RESULTS: Thirty-five participants completed the 12 week study. No significant changes in BW were noted although RM and WC dropped and LBM increased after training, particularly in the RTS group (FM 22.6 ± 5.5 kg to 21.2 ± 4.7 kg; LBM 68.5 ± 7.2 kg to 70.1 ± 7.4; p < 0.01). Subjects in the RTS group experienced more pronounced improvements in the strength measurements than the RT group. After the training intervention there were significant changes in hormonal and metabolic parameters as well as in glycemic control, particularly in the RTS group. CONCLUSIONS: Our data suggest that resistance training, particularly in combination with a soy protein based supplement improves body composition and metabolic function in middle aged untrained and moderately overweight males.

Effect of a supplement rich in alkaline minerals on acid-base balance in humans.

BACKGROUND: Western diets are considered acidogenic due to the high dietary acid load and a low intake of base-forming dietary minerals such as potassium, magnesium or calcium. In the present study we investigated the effect of a multimineral supplement (MMS) rich in alkaline minerals on acute and chronic regulation of acid-base balance with the pH of blood, urine and saliva as potential surrogate markers. METHODS: Parameters were measured (i) without MMS intake, (ii) in the three consecutive hours following ingestion (blood and urinary pH) and (iii) during one week with or without MMS intake (self-monitored using pH measurement strips). RESULTS: 25 (15 female; 10 male) subjects (age 44 +/- 14 y; BMI 23.9 +/- 1.9 kg/m2) were enrolled in the investigation. Following acute administration of the MMS in the morning, blood ph (1 and 2 h after ingestion) rose from 7.40 to 7.41; p < 0.05, and also urinary pH 3 h after ingestion (5.94 to 6.57; p < 0.05) increased significantly. Following longer-term supplementation, both the increase in urinary pH in the morning and in the evening occurred within 1 day. Compared to pH values without the MMS, average pH in urine was 11% higher in the morning and 5% higher in the evening. Analyses of food records showed that the increase in urinary pH was not related to dietary change. CONCLUSION: Our results suggest that the ingestion of a multimineral supplement is associated with both a significant increase in blood and urinary pH. The health related consequences of this supplementation remain to be determined.