Bone mineral metabolism and different indices of skeletal health of Ladakhi women living at high altitude.

Objectives: High altitude possesses a great challenge for human survival owing to low oxygen tension and has been reported to cause bone deterioration among sojourns of high altitude. The bone health of Ladakhi women is investigated for the first time in this study. Methods: Dual energy X-ray absorptiometry of Ladakhi women and sea level women was done at the radius and calcaneus using EXA-3000 (Osteosys, Korea), followed by colorimetric and Enzyme Linked Immunosorbent Assay analysis of parameters regulating bone health. Results: There was no statistically significant difference between bone mineral density of Ladakhi women and sea level women at radius (P = 0.287) or calcaneus (P = 0.839). Almost similar cases of osteopenia were reported at both sites measured in the study among both groups. Two post-menopausal Ladakhi women however, had osteoporosis at the radius while 4 had osteoporosis at calcaneus. Significant increase in calcium levels with a decrease in intact parathyroid hormone and an increase in calcitonin levels were observed in Ladakhi women as compared to sea level women. Though there was no significant difference in 25-hydroxy vitamin D levels of both groups, a higher percentage of 25-hydroxy vitamin D deficiency (77% vs 23%) was observed in Ladakhi women as compared to sea level women. Estradiol levels were similar in both groups. Conclusions: The present study suggest that there is no significant relationship between high altitude living and bone mineral density among Ladakhi women.

Anti-Mullerian Hormone and Macrophage Migration Inhibitory Factor Determine the Reproductive Health of Ladakhi Women Residing at 3,500 m.

Shaw, Snigdha, Himashree Gidugu, Gopinath Bhaumik, Maramreddy Prasanna Kumar Reddy, Usha Panjwani, and Dishari Ghosh. Anti-Mullerian hormone and macrophage migration inhibitory factor determine the reproductive health of Ladakhi women residing at 3,500 m. High Alt Med Biol. 22:317-326, 2021. Background: Reproductive health of Ladakhi high-altitude (HA) native females was investigated for the first time in this study. Available literature suggest that, female reproductive cycle and hormonal profile varies in different HA populations due to heterogeneity. Although these studies illustrate some progress on the role of HA hypoxia, it still leaves scope for evaluation of the remaining mechanisms involved in the maintenance of reproductive health in this contemporary population. Materials and Methods: Menstrual details, phasic variations in circulatory steroid hormones, and gonadotropins along with oxytocin in sea level (SL) and HA (∼3,500 m) native females of India were assessed. Moreover, ovarian reserve marker anti-Mullerian hormone (AMH) and proinflammatory cytokine macrophage migration inhibitory factor (MIF) were measured. Results: A difference in Ladakhi women was registered compared to SL, regarding luteinizing hormone (LH) (2.6 mIU/ml vs. 4.4 mIU/ml, p < 0.05) and progesterone (P) (4.1 ng/ml vs. 9.4 ng/ml, p < 0.05) levels in their luteal phase. Reduced LH might contribute to poor development of the ovarian corpus luteum, subsequently diminish P level. Decreased AMH level in three age groups: 21-30 years (1.4 ng/ml vs. 3.2 ng/ml, p < 0.01), 31-40 years (0.6 ng/ml vs. 2.1 ng/ml, p < 0.01), and >40 years (0.4 ng/ml vs. 1.7 ng/ml, p < 0.01) of Ladakhi women were recorded than their SL counterpart. Elevated oxytocin (83.5 ng/ml vs. 76.3 ng/ml, p < 0.05) and MIF levels (70.2 ng/ml vs. 49.7 ng/ml, p < 0.01) along with low P and AMH levels delineated the reason for recorded early menopause (43.9 years), shorter reproductive span (∼29 years), and history of miscarriage in HA dwellers compared to SL. Conclusion: Therefore, the findings insinuated that the response of the reproductive system to hypoxia in Ladakhi women differs from SL women, and the adaptive response in these women might be in favor of their reproductive health.

Alterations of estrous cycle, 3ß hydroxysteroid dehydrogenase activity and progesterone synthesis in female rats after exposure to hypobaric hypoxia.

The underlying mechanism regulating hypoxia induced alteration in female steroid hormones is first time explored in this study. To understand the mechanistic approach, female Sprague- Dawley rats were exposed to acute and chronic hypobaric hypoxia (282 mm-Hg, ~7620 m, 6 hours, 3 and 7 days). Estrous cycle, body weight, plasma progesterone and estradiol levels, morphology, histology and two key steroidogenic enzymes: 3ß hydroxysteroid dehydrogenase (HSD) and 17ß HSD activity of ovary and adrenal gland were studied. A persistent diestrous phase and a significant decrease in body weight were found in chronic hypoxia groups. Histological study suggested degenerative changes in ovarian corpus luteum of 7 days chronic hypobaric hypoxia (7CHH) group and a declined percentage of adrenocortical cells in 3 days chronic hypobaric hypoxia (3CHH) and 7CHH groups. Plasma estradiol level was unaltered, but progesterone level was decreased significantly in all hypoxic groups. Ovarian 3ß HSD activity was decreased significantly with increasing days of hypoxic treatment along with a significantly low adrenal 3ß HSD activity in 7CHH. In conclusion, hypobaric hypoxia causes a state of low circulatory progesterone level in females likely due to the degenerative changes in the female ovarian and adrenal tissues together with low steroidogenic 3ß HSD enzyme activity.

Impact of high altitude on key determinants of female reproductive health: a review.

Imperishable research work was done on females visiting high-altitude (HA) areas for recreational activities or job purposes as well as on female HA natives. Hypoxia at HA is an unavoidable condition that affects the determinants of female reproductive functions like, the age of menarche and menopause, whole reproductive span, hormone synthesis, and fertility. This review will emphasize whether HA hypoxia is a threat to women: residents or visitors by analyzing these proximate determinants. Delayed menarcheal and advanced menopausal age was found to shorten the reproductive span in some HA populations, whereas in some cases, menstrual cycle was also reported to be irregular. In addition, the completed fertility rate (CFR) was increased when people migrated to lower altitude. Altered stress hormones and reproductive hormones were observed in sea-level females exposed to HA. Oxidative stress (OS) at HA was also reviewed to explain the probable reasons for the observed changes in these determinants because disturbed redox homeostasis may be a connecting link, affecting the reproductive functions. In conclusion, HA hypoxia plays a crucial role on various determinants of female reproductive health and this review will be helpful for more precise study along with the probable underlying mechanisms responsible for the changes in female reproductive functions at HA.

Post-translational modifications of eNOS augment nitric oxide availability and facilitates hypoxia adaptation in Ladakhi women.

The lower inhaled oxygen per volume at high altitude poses an intimidating challenge for humans to survive and reproduce. Indigenous populations of the Himalayas reportedly exhibit higher microcirculatory blood flow accompanied by higher orders of magnitude of nitric oxide (NO) products in lung, plasma and red blood cells as a vascular adaptation strategy for hypobaric hypoxia. The precise mechanism of such observed higher NO metabolites for hypoxia adaptation remains elusive. Studying high altitude native Ladakhi women, we observed significant higher eNOS mRNA and protein in blood/plasma as compared to lowland women. We also observed higher level of plasma l-citrulline and NOx (nitrates and nitrites) with concomitant lower levels of arginase mRNA and protein further suggesting higher eNOS activity and NO bioavailability. Interestingly, middle aged postmenopausal Ladakhi women exhibited significantly higher level of eNOS activity, NOx and cGMP as compared to age matched lowland women. Preferential phosphorylation of eNOS on stimulatory Ser1177 and Ser615 as well as dephosphorylation of inhibitory Thr495 site contributed to higher NO availability in Ladakhi women irrespective of age. We also observed higher levels of eNOS activating humoral factors like bradykinin and estrogen in both young and middle-aged Ladakhi women. These results suggest that an altered phosphorylation status, together with an enhanced expression of eNOS and potential humoral endothelial activators, are involved in enhanced activation of the eNOS-NO-cGMP pathway in Ladakhi women irrespective of age, reinforcing the hypothesis that NO metabolites play a major role in Himalayan pattern of hypoxia adaptation.

Elevated pulmonary artery pressure and brain natriuretic peptide in high altitude pulmonary edema susceptible non-mountaineers.

Exaggerated pulmonary pressor response to hypoxia is a pathgonomic feature observed in high altitude pulmonary edema (HAPE) susceptible mountaineers. It was investigated whether measurement of basal pulmonary artery pressure (Ppa) and brain natriuretic peptide (BNP) could improve identification of HAPE susceptible subjects in a non-mountaineer population. We studied BNP levels, baseline hemodynamics and the response to hypoxia (FIo2 = 0.12 for 30 min duration at sea level) in 11 HAPE resistant (no past history of HAPE, Control) and 11 HAPE susceptible (past history of HAPE, HAPE-S) subjects. Baseline Ppa (19.31 ± 3.63 vs 15.68 ± 2.79 mm Hg, p < 0.05) and plasma BNP levels (52.39 ± 32.9 vs 15.05 ± 9.6 pg/ml, p < 0.05) were high and stroke volume was less (p < 0.05) in HAPE-S subjects compared to control. Acute hypoxia produced an exaggerated increase in heart rate (p < 0.05), mean arterial pressure (p < 0.05) and Ppa (28.2 ± 5.8 vs 19.33 ± 3.74 mm Hg, p < 0.05) and fall in peripheral oxygen saturation (p < 0.05) in HAPE-S compared to control. Receiver operating characteristic (ROC) curves showed that Ppa response to acute hypoxia was the best variable to identify HAPE susceptibility (AUC 0.92) but BNP levels provided comparable information (AUC 0.85). BNP levels are easy to determine and may represent an important marker for the determination of HAPE susceptibility.

Alterations in different indices of skeletal health after prolonged residency at high altitude.

AIMS: The aim of this study was to evaluate the effect of prolonged residency at high altitude (HA) on different indices of bone health in sea level (SL) residents staying at an altitude of 3450 m for 4 months to 1 year. The assessment of bone health parameters included multisite quantitative bone speed of sound (SOS), and markers of bone metabolism such as serum alkaline phosphatase (ALP), bone specific alkaline phosphatase (BAP), urinary deoxypyridinoline (DPD), C-terminal propeptide of type I collagen (CICP), N-telopeptide of type I collagen (NTX), and hormonal regulators such as 25-hydroxy vitamin D3 (25Vit D), intact parathyroid hormone (i-PTH), and cortisol. RESULTS: The body weight in all the age groups was significantly lower at HA as compared to SL values. Prolonged residency at HA led to a significant decline in bone strength in terms of SOS, both at radius and phalanx. There was a significant increase in circulating Ca and ALP levels. Serum i-PTH and 25VitD levels decreased significantly. Significant decreases were also observed in CICP and BAP, bone formation markers, and serum NTX, DPD/Cr ratio, markers of bone resorption. CONCLUSION: These observations suggest that prolonged residency under hypoxic environment is associated with a decline in both bone formation and bone resorption markers, reflecting a lower bone turnover at HA.

Determination of bone mass using multisite quantitative ultrasound and biochemical markers of bone turnover during residency at extreme altitude: a longitudinal study.

A group of 221 male healthy volunteers of Indian Army were the subjects of the study. The baseline parameters of skeletal health were measured during their residency at an altitude of 3542 m. These subjects were then taken to an extreme altitude (EA, 5400-6700 m) where they stayed for about 4 months. The study parameters were repeated following their de-induction (DI) to 3542 m. On random selection, a subgroup was constituted from the above mentioned volunteers for detailed investigations on various bone turnover markers. Results of this study indicate a loss of body weight after DI from EA. The bone impairment was detected at the proximal phalanx, which is known to undergo early morpho-structural changes associated with bone resorption. The intact parathyroid hormone (i-PTH) levels showed a significant increase, while alkaline phosphatase (ALP) and bone specific alkaline phosphatase (BAP) activities declined significantly after DI from EA. This elevation in i-PTH might be required for maintenance of blood Ca level. 25 (OH) Vitamin D3 (25VitD) and calcitonin (CT) also showed a significant decline, which may suggest a negative impact on bone formation during sojourn at EA. The causes of deterioration of skeletal health at EA although are poorly understood but may be due to acute hypoxemia arising from extreme hypobaric hypoxia prevalent at extreme altitude.

Individual variation in response to simulated hypoxic stress of rats.

With an aim to categorize the animals exposed to simulated hypobaric hypoxia and to evaluate the hormonal profile responsible for individual variation in response to hypoxic stress, degree of tolerance to hypobaric hypoxia was measured by exposing the animals to a simulated altitude of 10,668 m at 32 degrees C and animals were categorized as low and high tolerant groups based on their gasping time. The hormonal profiles of these groups were evaluated just after exposure to the test. The results showed a distinct individual difference in response to hypoxic tolerance test. There was a significant increase in plasma norepinephrine concentration in high tolerant group than low tolerant rats. After hypoxic tolerance test, total circulating corticosterone (CORT) level also increased but this was not significant in high tolerant rats as compared to low tolerant rats. Corticosteroid binding globulin (CBG) concentration differ significantly between high and low tolerant groups of rats resulting in significant changes in circulating free corticosterone that in turn may be responsible for individual differences in hypoxic gasping time. Significant differences were also observed in prolactin and testosterone levels of both the groups. The results established the method of differentiating the animals according their response to hypoxic tolerance test. These data indicate that multiple components rather than only plasma glucocorticoid of the stress response are providing a basis for individual differences in physiological responses to hypoxic stress.

Bioavailability of iodine and hardness (magnesium and calcium salt) in drinking water in the etiology of endemic goitre in Sundarban delta of West Bengal (India).

Endemic goitre has been reported from the ecologically diverse Sundarban delta of West Bengal (India). To study the etiological factors for the persistence of endemic goitre, bioavailability of iodine and hardness of water used for drinking in the region were evaluated because these common environmental factors are inversely and directly related with goitre prevalence in several geographical regions. For the present study from 19 Community Development Blocks of Sundarban delta, 19 areas were selected at random. From each area at least 8 drinking water samples were collected and analyzed for iodine and the hardness (calcium and magnesium salt content). Iodine content in the drinking water samples was found in the range from 21 to 119 mg/L and total hardness of drinking water was found to range from 50 to 480 ppm. Presence of magnesium salt was found higher than the calcium salts in most of the samples. These findings suggest that the entire delta region is environmentally iodine sufficient but water is relatively hard and thus possibility of hardness of water for the persistence of endemic goitre may not be ruled out.

Effect of radish (Raphanus sativus Linn.) on thyroid status under conditions of varying iodine intake in rats.

Cruciferous plants viz. cabbage, cauliflower, turnip, radish, mustard etc. that contain goitrogenic/antithyroid substances, constitute a portion of regular human diet. The effect of chronic feeding of fresh and cooked radish, R. sativus under varying state of iodine intake on morphological and functional status of thyroid in albino rats was evaluated by thyroid gland morphology and histology, thyroid peroxidase activity, serum triiodothyronine, thyroxine and thyrotropin levels. The consumption pattern of iodine and goitrogens of cyanogenic origin was evaluated by measuring urinary iodine and thiocyanate levels respectively. After chronic radish feeding, increased weight of thyroid gland, decreased thyroid peroxidase activity, reduced thyroid hormone profiles and elevated level of thyrotropin were observed resembling a relative state of hypoactive thyroid gland in comparison to control even after supplementation of adequate iodine.

Goitre prevalence and the state of iodine nutrition in the sundarban delta of north 24-parganas in West Benegal.

The main objective of this study was to assess the iodine nutritional status among school children (6-12 yrs) of Sundarban delta in North 24-Parganas district of West Bengal in eastern India. A total of 2050 children were clinically examined for goiter and 240 urine samples were analyzed for iodine and thiocyanate respectively; iodine content in 48 water samples and 210 salt samples were also measured. Results indicate that the studied region is clinically severely goiter endemic having goiter prevalence 33.1 % (grade 1: 30.4 %; grade 2: 2.7%),median urinary iodine level 200 mug/l indicating no biochemical iodine deficiency, 65.2 % salt samples contain recommended level of iodine and the iodine content in drinking water is sufficient while mean urinary thiocyanate level was 0.708+/-0.38 mg/dl. In spite of the consumption of adequate iodine, the existing goiter prevalence may be for the consumption of dietary goitrogens/antithyroid substances that possibly come through food and water.

Evaluation of possible goitrogenic and anti-thyroidal effect of nitrate, a potential environmental pollutant.

Nitrate is a wide spread contaminant of ground and surface water. The source of nitrate in the ground water may be from run off or seepage from fertilized soil, municipal or industrial waste water, land fills, septic system, urban drainage or decaying plants. Human and animal systems are affected severely on nitrate exposure. The study was to investigate the effect of dietary nitrate exposure on the thyroid status along with the state of iodine nutrition. Rats were fed diet containing 3% potassium nitrate (KNO3) for 4 weeks and then thyroid status was evaluated by thyroid gland weight, urinary iodine excretion pattern, thyroid peroxidase (TPO) activity, serum levels of total thyroxine (T4), triiodothyronine (T3) and thyroid stimulating hormone (TSH) concentrations. In nitrate treated animals, the weight of thyroid gland was increased significantly (P<0.001) while thyroid peroxidase activity (P<0.01), serum T4 (P<0.01) and serum T3 levels (P<0.001) were reduced; but serum TSH level was increased (P<0.001) along with slightly elevated iodine excretion level (P<0.001) in comparison to control animals. The overall results indicated the development of a relative state of functional hypothyroidism with enlarged thyroid after nitrate exposure. This study can explain a part for the persistence of residual goitre in the post-salt iodization phase.

Iodine nutritional status & prevalence of goitre in Sundarban delta of South 24-Parganas, West Bengal.

BACKGROUND & OBJECTIVE: In post salt iodization phase endemic goitre and associated iodine deficiency disorders (IDD) were found prevalent in a randomly selected rural area of Sundarban delta and its adjoining areas of West Bengal. The present investigation was thus undertaken to study the total goitre rate, urinary iodine and thiocyanate excretion pattern of the school going children, iodine content in edible salt and drinking water in the Sundarban delta of South 24-Parganas in West Bengal. METHODS: A total of 4656 school children (6-12 yr) were clinically examined for goitre from 13 different areas in the delta region. Urinary iodine and thiocyanate levels were measured in 520 (40 from each area) samples collected randomly to evaluate the iodine nutritional status and consumption pattern of dietary goitrogen. Simultaneously iodine content was determined in 104 (8 from each area) drinking water samples and 455 (35 from each area) edible salt samples collected from the areas. RESULTS: Children of all the areas were affected by endemic goitre. The prevalence rates were in the ranges from 25-61 per cent; overall goitre prevalence was 38.2 per cent (grade 1--34.0%; grade 2--4.2%). Median urinary iodine level in the studied areas was 225 microg/l (range 115-525 microg/l) indicating no biochemical iodine deficiency in the region. Mean urinary thiocyanate levels were in the range from 0.326-1.004 mg/dl. Iodine content in drinking water samples were in the ranges from 22-119 microg/l, and 55.6 per cent edible salt samples had iodine level above the recommended 15 ppm at the consumption point. INTERPRETATION & CONCLUSION: The severity of endemic goitre was high in the studied population though the iodine nutritional status was found satisfactory in the region indicating no biochemical iodine deficiency. The people of the region consumed iodine through iodized salt but about 44 per cent of the salt samples at household level contained inadequate iodine, however their iodine intake was compensated through iodine in water and food. They also consumed dietary goitrogen. Environmental factors other than iodine deficiency may have possible role for the persistence of endemic goitre in the region. More investigations are thus necessary to arrive at certain definite cause of high goitre rates in this population.

Effect of bamboo shoot, Bambusa arundinacea (Retz.) Willd. on thyroid status under conditions of varying iodine intake in rats.

Young shoots or sprouts of common bamboos are used as food in third world countries. Evidences suggest the presence of cyanogenic glucoside like anti-thyroidal substance in bamboo shoots (BS) but effect of prolonged BS consumption on thyroid status under conditions of varying iodine nutriture remains unexplored. The study was undertaken to evaluate goitrogenic content, in vitro anti thyroid peroxidase (TPO) activity and in vivo anti thyroid potential of BS with and without extra iodide. Fresh BS contains high cyanogenic glucoside (551 mg/kg), followed by thiocyanate (24mg/kg) and glucosinolate (9.57mg/kg). In vitro inhibition in TPO activity was found with raw, raw boiled and cooked extracts. Inhibition constant (IC50) and PTU equivalence of fresh BS were 27.5+/-0.77 microg and 3.27 respectively. Extra iodide in the incubation media reduced TPO inhibition induced by BS but could not cancel it. Thyroid weight, TPO activity and total serum thyroid hormone levels of BS fed animals for 45 and 90 days respectively were determined and compared with controls. Significant increase in thyroid weight as well as higher excretion of thiocyanate and iodine along with marked decrease in thyroid peroxidase activity, T4 and T3 levels were observed in BS fed group. Chronic BS consumption gradually developed a state of hypothyroidism. Extra iodide had reduced the anti-thyroidal effect of BS to an extent but could not cancel it because of excessive cyanogenic glucoside, glucosinolate and thiocyanate present in it.