Monthly Archives: June 2021

Acclimation, Altitude Science, Genetics, Health, High Altitude, Medicine, Neonatal

Maternal Intermittent Hypoxia and the Effect on Adult Respiratory Control and the Gut Microbiome in Male Offspring

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On Friday, June 4th, I had the pleasure of attending the, online, Fifth Annual Center for Physiological Genomics of Low Oxygen (CPLGO) Summit. There were many great presentations that I had the opportunity to watch, including the presentation of Dr. Christine Ebert-Santos’ study looking at nighttime pulse oximetry in participants living at high altitude for longer than one year. The presentation this post will discuss is about research conducted at the University of Florida by Dr. Tracy Baker, PhD. This presentation was particularly of interest because it looks at hypoxia in relationship to Obstructive Sleep Apnea (OSA). OSA occurs when patency of the upper airway is compromised and air is inhibited from passing, leading to hypopnea and obstructive apneas1. Hypopnea are episodes of greater than 30% decrease in air flow that lasts ≥ 10 seconds with continued respiratory effort1. Obstructive apnea is a total stop in airflow that lasts ≥ 10 seconds with chest and abdominal efforts to continue breathing1. Patients with OSA have a higher apnea-hypopnea index (AHI) at altitude than at sea level, meaning that their time with decreased oxygenation while sleeping is increased at higher altitude2. Additionally, patients living at altitude with mild or moderate sleep apnea may have a false negative sleep apnea result when having a sleep study performed at sea level, which means that patients who have OSA at altitude will not show signs of sleep apnea at sea level, therefore missing the diagnosis on sleep study2.

Currently, it is understood that the effects of hypoxia secondary to sleep apnea takes a toll on the body over time. Patients often experience snoring and daytime sleepiness in addition to other symptoms or changes to the body that may not be as easily recognizable, such as living in an increased inflammatory state1,3. Further, it is well known that an adverse maternal environment during pregnancy can lead to long term fetal complications3. Combining these two concepts, Dr. Baker wanted to further investigate the adverse effects of hypoxia due to maternal sleep apnea to get a better understanding of the subsequent consequences this deprived oxygen state has on mothers and their offspring. The hypothesis of this study was: Intermittent hypoxia during pregnancy has detrimental and long-lasting consequences on offspring neural function.

To test this hypothesis, Dr. Baker and her team exposed pregnant rats to intermittent hypoxia during days 10-21 of gestation. OSA was modeled from hypoxia episodes, but not the sleep fragmentation that accompanies the disease. It is understood that both components of sleep apnea, hypoxia and sleep fragmentation, would have their own influence on the offspring. The rats were put into a chamber and delivered 15 episodes of hypoxia per hour, as people can experience up to 10-20 episodes of apnea per hour during pregnancy, in increments of 90 seconds with 90 seconds of normoxia in between. During hypoxic episodes, oxygen levels were brought down to 10% and oxygen saturation was reliably reduced to 85% with adequate re-saturation during episodes of normoxia. Control rats were exposed to normoxia, 21% oxygen “on and off”, to control for and take into consideration confounding factors such as air flow within the chamber. While in labor, the rats were then removed from the chamber to give birth in a normal environment. The baby rats, or pups, were never exposed to hypoxia after they were born. Lastly, the pups were followed into adulthood to monitor for long term effects.3

Next, data from the gestational induced hypoxia (GIH) offspring and the control rat offspring (GNX) was compared. The GIH offspring showed no evidence for obesity and no difference in the volume of fat pads from shortly after birth to 12 weeks, as they had the same trajectory as the GNX offspring. There was also no difference between gestational length, number of pups, pup retrieval time, or pup survival between the two groups. To evaluate effects of gestational hypoxia on breathing, the adult offspring were placed in a plethysmography chamber. A plethysmography chamber measures changes in volume in the body to assess how much air is in the lungs when breathing. The rats were given one-hour to acclimate in the chamber and ventilation was then measured over the following three hours. Of the two groups, male GIH offspring had a significantly increased number of spontaneous apneas per hour compared to male and female GNX offspring and female GIH offspring, who had no change in the number of apneic episodes. Apneic episodes are defined as a pause in breathing that lasts longer than the duration of two breaths. Spontaneous apneic episodes are episodes of apnea with no apparent trigger on plethysmography signal. Approximately 60% of GIH male offspring had spontaneous apnea out of the 95% confidence interval, suggesting gestational intermittent hypoxia altered the phenotype in the male offspring. Again, this was not a congruent finding in GIH females.3

An additional factor to be considered in this scenario is respiratory plasticity, which is the body’s ability to help animals adapt to life changing circumstances, such as hypoxia and sleep apnea. A body’s ability to have respiratory plasticity is suggestive of a healthy neural system because breathing is an automatic and rhythmic function of the brain stem. Ultimately, the respiratory system you are born with is not the one you will die with. Recurrent central apnea can promote respiratory plasticity. Dr. Baker’s team further investigated whether the GIH rats had an altered adaptive response to conditions that alter the body’s natural response to breathing, which in this case is recurrent central apnea. Her team mechanically ventilated adult offspring that were vagalized, paralyzed, and urethane anesthetized to study neural control of breathing independent from the process of ventilation, and data was recorded via phrenic neurograms. A neural apnea was caused by lowering PaCO2 levels lower than the level for breathing and was then stopped by raising PaCO2 back to baseline levels. Recurrent neural apneas triggered plasticity mechanisms to make it harder to elicit the next apnea. Data showed adult male GIH offspring have impaired responses to recurrent reductions in respiratory neural activity and did not express plasticity following a triggered central apnea episode. Like prior results, female GIH offspring did not have this same neural plasticity impairment as the males, showing no elevations in spontaneous apnea and intact compensatory plasticity triggered by central apneas.3

Further, adult offspring were assessed for increased inflammation. To no surprise, GIH males had increased basal neuroinflammation. Although both male and female GIH offspring had increased inflammatory markers, the females were able to suppress the inflammation by an unknown mechanism that the male GIH offspring could not. Adult offspring of GIH and GNX groups were exposed to bacterial lipopolysaccharide (LPS), which confirmed that the GIH males mounted a greater inflammatory response compared to the other offspring, suggesting these males have an altered inflammation response. In the central nervous system (CNS), microglia are innate immune cells that can produce inflammatory cytokines and comprise 5-10% of CNS cells. Dr. Baker’s team pharmacologically depleted these cells in the adult offspring by administering the drug PLX3397 for seven days. This resulted in a stark reduction of microglia by 86%. The GIH male offspring with depleted microglia were able to regain compensatory plasticity triggered by recurrent central apneas. Three days after stopping PX3397, the microglia came back and expanded. When the microglia repopulated, there was restoration of the impaired plasticity phenotype in GIH males.3

To get a better understanding of what could be driving the persistent microglial inflammation in the GIH males, the gut-brain-axis was assessed. In human literature, it is suggested that sleep apnea is associated with gut dysbiosis. Investigating this link, feces was collected from the rats which showed diversity in the bacterial species present in GIH males compared to GIH females and both sexes of the GNX group. Dr. Mangalam looked at the GI bacteria shift and determined the gut microbiomes were comprised of two main phyla of bacteria, Bacteroidetes and Firmicutes. GIH males had increased Bacteroidetes and decreased Firmicutes compared to the other offspring. Initially unsure of this significance, Dr. Mangalam deduced that the decreased bacteria in the GIH male microbiome produce a short string fatty acid called butyrate. Once produced, this fatty acid stimulates the release of neuropeptides and serotonin, which are up taken by the portal vein. From there, butyrate enters the blood circulation and crosses the blood brain barrier (BBB), stimulating active receptors on the vagus nerve. Butyrate supports plasticity in the brain and reduces inflammation.3

This leads to the final question: can the neural plasticity deficit be rescued by decreasing neuroinflammation by supplementing male GIH offspring with butyrate? GIH male rats were supplemented with eight doses of 2mg/kg of Tributyrin over 22 days, which is converted into butyrate3. Upon creating central apnea in the GIH males treated with Tributyrin, it was found that their respiratory plasticity was fully rescued3. So, what does this mean?

Simply, this means gestational intermittent hypoxia has sex-specific, long-lasting effects on adult offspring physiology. This is shown by: 1) gut dysbiosis in male offspring, 2) increased central apneas during sleep with impaired respiratory plasticity, 3) enhanced basal inflammation of microglia in male offspring with increased inflammatory response upon provocation, and 4) microglial depletion or butyrate supplementation repaired deficits in respiratory plasticity.3

The research conducted by Dr. Baker’s team opens additional research opportunities regarding effects of hypoxia on vulnerable populations, such as pregnant mothers and their offspring. The findings from this study can be retested and built upon as research continues to be done. Although this research was not conducted at altitude, it is still interesting and pertinent to the altitude community, as hypoxia and OSA are common problems at altitude. This study contributes important knowledge to the science and medical community; however, more research will need to be done to confirm and fully understand the adverse effects of hypoxia during pregnancy. Further, more information is needed to understand how effects of gestational hypoxia can be applied to populations experiencing hypoxia secondary to living at altitude in a low oxygen environment.

References:

  1. Guilleminault C, Zupancic M. Sleep Disorders Medicine. Third Edition. Philadelphia, PA. Saunders. 2017. pp: 319-339.
  2. Patz D, Spoon M, Corbin R, et al. The effect of altitude descent on obstructive sleep apnea. CHEST. 2006; 130(6): 1744-1750. Doi: https://doi.org/10.1378/chest.130.6.1744
  3. Baker T. CoBAD: Maternal Intermittent Hypoxia and the Effect on Adult Respiratory Control and the Gut Microbiome in Male Offspring. Oral presentation at: the Fifth Annual Center for Physiological Genomics of Low Oxygen (CPLGO) Summit; June 4th, 2021; online.

Amanda Smith is a second year PA student at Drexel University in Philadelphia, Pennsylvania. Amanda was raised in the “sweetest place on Earth”, Hershey, Pennsylvania. She obtained her B.S. in Health Science with a double minor in Creative Writing and Community Health at Hofstra University on Long Island. Between obtaining her undergraduate and graduate degrees, Amanda worked as an Emergency Department scribe, pediatric nurse aide, and as a lead research coordinator in Neurosurgery/Neuro-Oncology at the Penn State Hershey Neuroscience Institute. Amanda loves to travel and was able to incorporate her love for traveling and medicine by traveling across the country for clinical rotations, rotating at sites in New York, California, Pennsylvania, and Colorado, with her next destination in Alaska!

Acclimation, Altitude Science, Health, High Altitude, Mental Health, Mountaineering

Anxiety at Altitude

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As I arrived in Denver (5280′), and ultimately Frisco, CO (9000′), the first physical symptom I noticed from the high-altitude environment was dyspnea on exertion. On flat ground I didn’t feel any different than at home in New Jersey, but as soon as I began to climb stairs or hike the beautiful trails in the area, I quickly became winded. I had already read about the common symptoms of high-altitude acclimation and knew this was normal and was on the lookout for headache, nausea, or dizziness. I noticed my resting heart rate was elevated and told myself this was also normal because the low-oxygen environment required my heart to work harder to keep my pulse oxygen levels up. I already owned a pulse oximeter that I had bought during my time working with COVID patients in the Emergency Room on a previous rotation. I checked that within my first week and was initially disappointed that it was averaging around 91%, but soon found out this was also normal, especially since I was still acclimating. My Apple watch trended data over time on my heart rate and I noticed a tremendous difference in my resting HR compared to home.

I landed on May 2nd, 2021 and I think the graphs above make that quite evident. My walking HR was even more noticeably elevated. “The initial cardiovascular response to altitude is characterized by an increase in cardiac output with tachycardia … after a few days of acclimatization, cardiac output returns to normal, but heart rate remains increased”1

My persistently elevated heart rate caused me to feel anxious when hiking or doing other physical activity, and that anxiety in turn raised my heart rate even more. I had experienced PVC’s in the past which occurred only a few times a month, nowhere near the threshold for treatment, and had been reassured they were totally benign. On a hike during my first week in Colorado I experienced a few of these “skipped beats” followed by rapid heart rate and had to talk myself down from the anxiety it caused. This is what prompted me to research the effect of altitude on anxiety. “Adrenergic centers in the medulla are activated in acute hypoxia and augment the adrenergic drive to the organs.”2 It seems as though the body’s compensatory mechanisms to physiological changes can be accompanied by unwanted mental health disturbances. This is especially true for people in the early stages of shifting from low altitude to high altitude. During the adjustment period individuals are most susceptible to new-onset anxiety disorders, but even those living long-term at high altitude are at increased risk of psychiatric ailments.4 In fact, living in high-altitude environments has been associated with serious mental health implications not limited to anxiety disorders, including depression and increased suicidality.4 This has been evidenced by statistically significant changes in PHQ-9 Total Score, PHQ-9 suicidal ideation, and GAD-7 Total Score.4

Sleep disturbances have often been faulted for these increases in anxiety and depression at high altitude, and although I didn’t have any formal sleep studies done while in Colorado, I felt well-rested and didn’t notice a change in my sleep at altitude.3 One hypothesis that could explain these findings and my personal experience, is that hypoxia has an inverse relationship with serotonin.4 Because oxygen is a requirement for the creation of serotonin, living somewhere with decreased oxygen could lead to deficiency. Serotonin has an expansive role in the human body, playing a role in cognition, sleep, mood, digestion and other crucial aspects of life. Low levels of this neurotransmitter have been implicated as a cause for depression and accordingly many of our best antidepressant medications like SSRI’s and SNRI’s work on these pathways. There is also a “chicken or the egg?” argument to be made. Is the anxiety brought on due to hypoxia which in turn causes somatic symptoms like palpitations, shortness of breath, and presyncope; or do these symptoms caused by hypoxia come first, resulting in anxiety and panic attacks? For example, hyperventilation, a well-known provocative factor of panic attacks, is also a response to altitude changes. Hypoxia leads to hypocapnia, which can ultimately lead to respiratory alkalosis.5 Although there are multiple hypotheses for these mental health changes, there does seem to be an agreement in the literature that they do exist.

Luckily, in my experience, my body adjusted over the span of a few weeks. My HR began to trend down towards my normal resting rate in the 70’s and my anxiety levels also dropped. I started doing more challenging hikes, traveling and enjoying the many natural wonders Colorado has to offer. Just being in amazing places like Rocky Mountain National Park and the San Isabel National Forest had a profound impact on my mood as I soaked in the scenery. I took pictures, breathed fresh mountain air and spotted wildlife, which all served to distract me from my worries. The mood-altering benefits of exercise also likely played a role in my increasing happiness. I grew to love the state and as soon as I felt fully adjusted, it was time to go back to New Jersey. Back to sea-level, outrageous humidity and hotter weather.

References:

  1. Naeije R. Physiological adaptation of the cardiovascular system to high altitude. Prog Cardiovasc Dis. 2010 May-Jun;52(6):456-66. doi: 10.1016/j.pcad.2010.03.004. PMID: 20417339.
  2. Richalet JP. Physiological and Clinical Implications of Adrenergic Pathways at High Altitude. Adv Exp Med Biol. 2016;903:343-56. doi: 10.1007/978-1-4899-7678-9_23. PMID: 27343107.
  3. Bian SZ, Zhang L, Jin J, Zhang JH, Li QN, Yu J, Chen JF, Yu SY, Zhao XH, Qin J, Huang L. The onset of sleep disturbances and their associations with anxiety after acute high-altitude exposure at 3700 m. Transl Psychiatry. 2019 Jul 22;9(1):175. doi: 10.1038/s41398-019-0510-x. PMID: 31332159; PMCID: PMC6646382.
  4. Kious BM, Bakian A, Zhao J, Mickey B, Guille C, Renshaw P, Sen S. Altitude and risk of depression and anxiety: findings from the intern health study. Int Rev Psychiatry. 2019 Nov-Dec;31(7-8):637-645. doi: 10.1080/09540261.2019.1586324. Epub 2019 May 14. PMID: 31084447.
  5. Roth WT, Gomolla A, Meuret AE, Alpers GW, Handke EM, Wilhelm FH. High altitudes, anxiety, and panic attacks: is there a relationship? Depress Anxiety. 2002;16(2):51-8. doi: 10.1002/da.10059. PMID: 12219335.

Joseph Albanese is a second-year physician associate (PA) student attending Drexel University in Philadelphia, Pennsylvania. He grew up in Hillsborough, NJ. He got his BA from The Pennsylvania State University as a double major in Psychology and Film Studies. Prior to PA school he worked as a mental health associate in an inpatient psychiatry setting with actively suicidal and homicidal patients. The acuity of the unit he worked on made him appreciate the benefits of talk-therapy, but also the crucial role of medicine in many cases. This led him to apply to PA school. In his free time Joe loves to travel (favorite places include Japan, Iceland, Glacier National Park, and now Colorado). He also enjoys photography, playing sports, and eating new foods.