Reduced Cerebrovascular Oxygenation in Individuals with Post-Acute COVID-19 Syndrome (PACS) ("long COVID").

There is evidence that hypoxia occurs in the brain of some individuals who contracted the COVID-19 disease. Furthermore, it has been widely reported that about 13% of individuals who contracted the COVID-19 disease report persistent symptoms after the acute infection stage (>2 months post-acute infection). This is termed post-acute COVID-19 syndrome (PACS) or ("long COVID"). In this study, we aimed to determine if hypoxia measured non-invasively with frequency domain near-infrared spectroscopy (fdNIRS) occurs in asymptomatic and symptomatic individuals with post-acute COVID-19 disease. We show that 26% of our symptomatic group, measured on average 9.6 months post-acute COVID-19 disease, were hypoxic and 12% of the asymptomatic group, measured on average 2.5 months post-acute infection, were hypoxic. Our study indicates that fdNIRS measure of hypoxia in the brain may be a useful tool to identify individuals that are likely to respond to treatments targeted at reducing inflammation and improving oxygenation.

Non-invasive Detection of Persistent Cortical Hypoxia in Multiple Sclerosis Using Frequency Domain Near-Infrared Spectroscopy (fdNIRS).

There may be a relationship between hypoxia and inflammation, which is important in the outcomes of a wide array of human diseases. Multiple sclerosis (MS) is one such disease. There is evidence that hypoxia may influence inflammation in MS. We showed previously that about 40% of participants with MS had hypoxia in the cortical grey matter using frequency-domain near-infrared spectroscopy (fdNIRS). In this study, we aimed to determine if hypoxia in MS persists chronically (for a year or more) by measuring at baseline and ≥12 months later. We found that hypoxia persists for at least a year in 80% of participants with MS. As more individuals remained hypoxic than returned to normoxia, the development of hypoxia may relate to disease progression.

Near-Infrared Spectroscopy Reveals Brain Hypoxia and Cerebrovascular Dysregulation in Primary Biliary Cholangitis.

BACKGROUND AND AIMS: Primary biliary cholangitis (PBC) is an autoimmune cholestatic liver disease linked to symptoms including fatigue and altered mood/cognition, indicating that chronic liver inflammation associated with PBC can impact brain function. We employed near-infrared spectroscopy (NIRS), a noninvasive neuroimaging technique, to determine whether patients with PBC exhibit reduced cerebral oxygen saturation (StO2 ) and altered patterns of microvascular cerebral blood perfusion and whether these alterations were associated with clinical phenotype. This observational case-control study was conducted at a tertiary hospital clinic (University of Calgary Liver Unit). APPROACH AND RESULTS: Thirteen female patients with noncirrhotic PBC, seven female patients with cirrhotic PBC, and 11 healthy female controls were recruited by physician referral and word of mouth, respectively. NIRS was used to measure cerebral hemoglobin and oxygen saturation. A wavelet phase coherence method was used to estimate the coherent frequency coupling of temporal changes in cerebral hemodynamics. The PBC group demonstrated significantly reduced cerebral StO2 (P = 0.01, d = 0.84), indicating cerebral hypoxia, significantly increased cerebral deoxygenated hemoglobin concentration (P < 0.01, d = 0.86), and significantly reduced hemodynamic coherence in the low-frequency band (0.08-0.15 Hz) for oxygenated hemoglobin concentration (P = 0.02, d = 0.99) and total hemoglobin (tHb) concentration (P = 0.02, d = 0.50), indicating alterations in cerebrovascular activity. Complete biochemical response to ursodeoxycholic acid (UDCA) therapy in early patients with PBC was associated with increased cerebral tHb concentration and decreased hemodynamic coherence. CONCLUSIONS: Using NIRS, patients with PBC were found to have hypoxia, increased cerebral hemoglobin concentration, and altered cerebrovascular activity, which were reversed in part in UDCA responders. In addition, symptoms and quality-of-life measures did not correlate with brain hypoxia or cerebrovascular dysregulation in patients with PBC.

Intra-articular injection of synovial mesenchymal stem cells improves cartilage repair in a mouse injury model.

Controversy remains whether articular cartilage has an endogenous stem/progenitor cell population, since its poor healing capacity after injury can lead to diseases such as osteoarthritis. In the joint environment there are mesenchymal stem/progenitor cells (MSCs) in the synovial membrane and synovial fluid that can differentiate into cartilage, but it is still under debate if these cells contribute to cartilage repair in vivo. In this study, we isolated a Sca-1 positive, chondrogenesis capable population of mouse synovial MSCs from C57BL6 and MRL/MpJ "super-healer" strains. Intra-articular injection of Sca-1 + GFP + synovial cells from C57BL6 or MRL/MpJ into C57BL6 mice following cartilage injury led to increased cartilage repair by 4 weeks after injury. GFP expression was detected in the injury site at 2 weeks, but not 4 weeks after injury. These results suggest that synovial stem/progenitor cells, regardless of strain background, have beneficial effects when injected into an injured joint. MSCs derived from MRL/MpJ mice did not promote an increased repair capacity compared to MSCs derived from non-healing C57BL6 controls; however, MRL/MpJ MSCs were observed within the defect area at the time points examined, while C57BL6 MSCs were not.

Evaluating endogenous repair of focal cartilage defects in C57BL/6 and MRL/MpJ mice using 9.4T magnetic resonance imaging: A pilot study.

The use of magnetic resonance imaging (MRI) for evaluating joint injuries is often considered superior to radiography due to the capacity of MRI for visualizing both soft and hard tissues. While longitudinal studies regarding cartilage repair have been undertaken on patients and in larger animal models, a method has yet to be developed for mouse cartilage to be repeatedly and non-invasively evaluated over time. The aim of this pilot study was to investigate if morphological changes following a focal cartilage injury in mice could be measured by 9.4T magnetic resonance imaging. Focal cartilage defects were induced in the left knee of 4-6weeks old C57BL/6 and MRL/MpJ mice. At endpoints 0, 2, and 4weeks post-injury, legs were dissected out and imaged ex vivo. The defect could be detected by MRI immediately after injury, appearing as a hyperintense focal point and with size similar to that of the surgical tool used. Defects were visible in both strains up to 4weeks post-injury, although signal intensity decreased over time. One C57BL/6 in particular, displayed extensive fibrosis in the patellar tendon at 4weeks as assessed by histology, while the MR images of the same animal displayed a clear, structural distinction between the patella and the new tissue growth. Overall, our results suggest that MRI could be used for longitudinal studies in murine cartilage injury models to evaluate certain characteristics of repair not detectable through histology.

A transmissive laser speckle imaging technique for measuring deep tissue blood flow: an example application in finger joints.

BACKGROUND AND OBJECTIVE: Laser speckle perfusion imaging (LSPI) is a minimally invasive optical measure of relative changes in blood flow, providing real-time, high resolution, two-dimensional maps of vascular structure. Standard LSI imaging uses a light-reflective geometry that limits the measurement to a thin surface layer of 0.2-1 mm. The objective of this study was to test a new LSI instrument geometry with the laser source opposed to the image capture plane (light transmissive). Captured light then travels the entire tissue thickness (10-15 mm), sampling much deeper regions of interest than conventional optical imaging techniques. STUDY DESIGN: Reflective-light (conventional) and transmissive-light LSI modes were used to measure finger joint blood flow during a timed tourniquet occlusion of the brachial artery in volunteer participants. RESULTS: There was greatly increased visibility of vessels underlying the skin in the light-transmissive mode LSI mode. Established LSI algorithms were shown to still work in the light-transmissive mode, despite decorrelation due to finite laser coherence length and the light passing through a tissue thickness of 10-15 mm. CONCLUSION: Transmissive LSI can be used to measure blood flow deep (10-15 mm) into tissues. This could be useful for non-invasive measurements of finger joint synovial blood flow in diagnosing and treating peripheral vascular disorders, such as rheumatoid arthritis.

Measuring oxygenation in vivo with MRS/MRI--from gas exchange to the cell.

Oxygen plays a major role as a substrate in metabolic processes in numerous signaling pathways, in redox metabolism, and in free radical metabolism. To study the role of oxygen in normal and pathophysiological states, methods that can be used noninvasively are required. This review examines the potential of nuclear magnetic resonance techniques to study tissue oxygenation. It is written from a systems perspective, looking at detection methods with respect to the path that oxygen takes in the mammalian system-from the lungs, through the vascular system, into the interstitial space, and finally into the cell. Methods discussed range from those that are quantifiable, such as the assessment of spin lattice relaxation time in fluorocarbon solutions, to those that are more correlative, such as assessment of lactate and high energy phosphates. Since the methods vary in their site of application, sensitivity, and specificity to the quantification of oxygen, this review provides examples of how each method has been applied. This may facilitate the reader's understanding of how to optimally apply different methods to study specific biomedical problems.

The effects of ketamine-xylazine anesthesia on cerebral blood flow and oxygenation observed using nuclear magnetic resonance perfusion imaging and electron paramagnetic resonance oximetry.

Ketamine-xylazine is a commonly used anesthetic for laboratory rats. Previous results showed that rats anesthetized with ketamine-xylazine can have a much lower cerebral partial pressure of oxygen (P(t)O(2)), compared to unanesthetized and isoflurane anesthetized rats. The underlying mechanisms for the P(t)O(2) reduction need to be elucidated. In this study, we measured regional cerebral blood flow (CBF) using nuclear magnetic resonance (NMR) perfusion imaging and cortical P(t)O(2) using electron paramagnetic resonance (EPR) oximetry in the forebrain of rats under isoflurane, ketamine, ketamine-xylazine and isoflurane-xylazine anesthesia. The results show that in ventilated rats ketamine at a dose of 50 mg/kg does not induce significant changes in CBF, compared to isoflurane. Ketamine-xylazine in combination causes 25-65% reductions in forebrain CBF in a region-dependent manner. Adding xylazine to isoflurane anesthesia results in similar regional reductions in CBF. EPR oximetry measurements show ketamine increases cortical P(t)O(2) while xylazine decreases cortical P(t)O(2). The xylazine induced reduction in CBF could explain the reduced brain oxygenation observed in ketamine-xylazine anesthetized rats.

Noninvasive assessment of cerebral oxygenation during acclimation to hypobaric hypoxia.

Factors regulating cerebral tissue PO2 (PtO2) are complex. With the increased use of clinical PtO2 monitors, it has become important to elucidate these mechanisms. The authors are investigating a new methodology (electron paramagnetic resonance oximetry) for use in monitoring cerebral PtO2 in awake animals over time courses of weeks. The authors used this to study cerebral PtO2 in rats during chronic acclimation to hypoxia predicting that such acclimation would cause an increase in PtO2 because of increases that occur in capillary density and oxygen carrying capacity. The average PtO2 between 7 and 21 days was increased by 228% over controls.

Critical oxygen tension in rat brain: a combined (31)P-NMR and EPR oximetry study.

The relationship between cerebral interstitial oxygen tension (Pt(O(2))) and cellular energetics was investigated in mechanically ventilated, anesthetized rats during progressive acute hypoxia to determine whether there is a "critical" brain Pt(O(2)) for maintaining steady-state aerobic metabolism. Cerebral Pt(O(2)), measured by electron paramagnetic resonance oximetry, decreased proportionately to inspired oxygen fraction. (31)P-nuclear magnetic resonance measurements revealed no changes in P(i), phosphocreatine (PCr)/P(i) ratio, or intracellular pH when arterial blood oxygen tension (Pa(O(2))) was reduced from 145.1 +/- 11.7 to 56.5 +/- 4.4 mmHg (means +/- SE). Intracellular acidosis, a sharp rise in P(i), and a decline in the PCr/P(i) ratio developed when Pa(O(2)) was reduced further to 40.7 +/- 2.3 mmHg. The corresponding Pt(O(2)) values were 15.1 +/- 1.8, 8.8 +/- 0.4, and 6.8 +/- 0.3 mmHg. We conclude that over a range of decreasing oxygen tensions, cerebral oxidative metabolism is not sensitive to oxygen concentration. Oxygen becomes a regulatory substrate, however, when Pt(O(2)) is decreased to a critical level.

Effect of hyperventilation on brain tissue oxygenation and cerebrovenous PO2 in rats.

Previous studies have shown that cortical tissue oxygenation is impaired during hyperventilation. However, it is important to quantify the effect of hyperventilation on brain tissue PO(2) and cerebrovenous PO(2) simultaneously especially since cerebral venous oxygenation is often used to assess brain tissue oxygenation. The present study was designed to measure the sagittal sinus PO(2) (PvO(2)), brain tissue PO(2) in the thalamus (PtO(2)), and brain temperature (Bt) simultaneously during acute hyperventilation. Isoflurane-anesthetized rats were hyperventilated for 10 min during which time the arterial carbon dioxide tension (PaCO(2)) dropped from 40.3+4.9 mmHg to 23.5+2.8 mmHg. PtO(2) declined from 26.0+/-4.2 mmHg to 14.8+/-5.2 mmHg (P=0.004) while brain temperature decreased from 36.5+0.3 degrees C to 36.2+0.3 degrees C (P=0.02). However, PvO(2) and arterial blood pressure (BP) did not change during hyperventilation. The maintenance of PvO(2) when perfusion is thought to decline and PtO(2) decreases suggests that there may be a diffusion limitation, possibly due to selective perfusion. Therefore, cerebrovenous PO(2) may not give a good assessment of brain tissue oxygenation especially in conditions of acute hyperventilation, and deeper brain regions other than the cortex also show impaired tissue oxygenation following hyperventilation.

In vitro MR imaging of hyaline cartilage: correlation with scanning electron microscopy.

OBJECTIVE: Our goal was to determine how the three-dimensional structure of hyaline cartilage affects its MR appearance and to correlate this appearance with detailed structural analysis using scanning electron microscopy and freeze-fracture sectioning techniques. MATERIALS AND METHODS: In vitro 7-T spin-echo MR images of hyaline cartilage specimens from four patients undergoing above-knee amputations were obtained parallel and perpendicular to the main magnetic field. Specimens were imaged with low- and high-power scanning electron microscopy after freeze fracturing. The corresponding images from both techniques were analyzed with specific attention to the three-dimensional structure of the cartilage, collagen fibril orientation, and respective changes in the MR appearance. RESULTS: Freeze fracturing of cartilage reveals a curved fracture plane. Expected changes in signal intensity predicted by the magic angle effect correlated with observed changes in signal intensity across the thickness of the sample. Changes in individual collagen fibril orientation did not correspond to MR layering. CONCLUSION: The three-dimensional organization of collagen in cartilage has a strong influence on the MR appearance of cartilage. This influence is caused by the restriction of water mobility and the resulting magic angle effect caused by curvature of the collagen network, possibly because of the influence on proteoglycan orientation.

Quantitative magnetic resonance imaging of the mdx mouse model of Duchenne muscular dystrophy.

The mdx mouse has a genetically homologous disease to Duchenne muscular dystrophy in humans. The disease progression, however, is not accompanied by the same level of fatty infiltration and muscle degeneration as occurs in humans. Thus, the presence of histological/pathological changes in living mdx mice has been difficult to monitor. We quantified proton density and the T2 relaxation time of protons with a resolution of 195 x 195 x 1000 microm using multiecho magnetic resonance (MR) imaging at 7 Tesla. These relaxation data were correlated with water content in both muscle and brain of mdx and controls. No differences were observed in brain. The mdx muscles had increased water content and proton density and decreased T2 relative to controls. These data indicate that there are intrinsic changes in T2 (opposite to that which would be induced by fatty infiltration) and suggest that T2 imaging could be used to monitor progression and treatment in this animal model.

Altered brain metabolism in the C57BL/Wld mouse strain detected by magnetic resonance spectroscopy: association with delayed Wallerian degeneration?

In the C57BL/Wld(s) (Wld) mouse strain, both PNS and CNS axonal disintegration during Wallerian degeneration is dramatically slowed, with isolated axons being able to conduct compound action potentials (CAPs) for several weeks post-transection. The ability to conduct a CAP signifies the presence of an intact plasma membrane, normal ion gradients, and functioning ion channels. In neurons, ion homeostasis is primarily regulated by the Na(+)-K(+)-ATPase, which utilizes approximately 50% of neuronal energy output. To investigate the possibility that the Wld mutation prolongs axonal degeneration by conferring a more favorable energetic status to neurons or alters metabolism, we used 31P and 1H magnetic resonance spectroscopy (MRS) to compare the cerebral and muscle energy metabolism, membrane phospholipid contents, and water-soluble metabolites of Wld and wild-type (C57BL/6J [6J], and BALB/c) mouse strains. We first demonstrate that, with advancing age, transected Wld CNS nerves degenerate faster, paralleling previous findings in the PNS. We found significantly decreased phosphocreatine and phosphomonoester concentrations in the brains of Wld mice at 1- and 2-months of age compared to both 6J and BALB/c mice, but we failed to find differences in the adenylate (ATP, ADP, or AMP) or phospholipid concentrations. In another excitable tissue, skeletal muscle, no differences in energy-containing metabolites were detected. High resolution 1H MRS indicated that at 1 month of age, Wld brains have cytosolic levels of glutamate and phosphocholine that are significantly decreased, relative to total N-acetyl aspartate content. Our results demonstrate that delayed Wallerian degeneration in the C57BL/Wld mouse strain is associated with altered cerebral metabolism, although these changes may be secondary to the mutation.

Regional heterogeneity in the brain's response to hypoxia measured using BOLD MR imaging.

Blood oxygenation level-dependent (BOLD) magnetic resonance (MR) imaging is sensitive, in part, to the amount of paramagnetic deoxyhemoglobin in a voxel. This project was designed to determine whether there would be differences in the BOLD response between the hippocampus and other brain regions to acute hypoxia. R2* was quantified using a multi-echo gradient-echo sequence. The pyramidal CA1 region of the hippocampus showed a reduced response to changes in arterial oxygenation relative to cortex and basal ganglia and white matter. This difference may relate to the relative hypoxia sensitivity of the hippocampus. It also supports the premise that in functional MR imaging, the magnitude of the MR response to a stimulus may vary with the region of the brain.

Classification of biopsy-confirmed brain tumors using single-voxel MR spectroscopy.

BACKGROUND AND PURPOSE: Our purpose was to develop a classification scheme and method of presentation of in vivo single-voxel proton spectroscopic data from astrocytomas that most closely match the classification scheme determined from biopsy specimens. Since in vivo proton spectroscopy is noninvasive, it may be an attractive alternative to intracranial biopsy. METHODS: Single-voxel spectra were acquired using the point-resolved spectroscopic pulse sequence as part of the Probe spectroscopy package on a G.E. 1.5-T Signa scanner. Subjects consisted of 27 patients with biopsy-confirmed brain tumors (13 with glioblastoma multiforme, six with anaplastic astrocytoma, and eight with low-grade astrocytoma). The patients were divided into groups based on the histologic subtype of their tumor for different treatment protocols. RESULTS: Metabolic peak areas were normalized for each metabolite (choline, creatine, N-acetylaspartate, lactate) to the area of the unsuppressed water peak and to the area of the creatine peak. Kruskal-Wallis nonparametric analysis of variance (ANOVA) tests showed statistically significant differences among the tumor groups for all the area ratios. The lactate/water ratio could be used to distinguished all three tumor groups, whereas the choline/water ratio distinguished low-grade astrocytomas from the two high-grade groups. Both the choline and lactate ratios could be used to separate the high-grade from the low-grade tumors. CONCLUSION: Specific relative metabolic peak area ratios acquired from regions of contrast-enhancing brain tumor can be used to classify astrocytomas as to histopathologic grade.

Measurement of arterial, venous, and interstitial pO2 during acute hypoxia in rat brain using a time-resolved luminescence-based oxygen sensor.

This is the first publication using a fiber optic "optode" and a luminescence based pO2 detection method for assessing neural tissue oxygenation. The system was used to simultaneously monitor pO2 in tissue (PtO2) and venous blood (PvO2) during normoxia, hyperoxia and hypoxia. PaO2 was varied by changing inspired oxygen (FiO2) from 0.3 to 0.13. Tissue and arterial pO2 were measured in 5 rats while the simultaneous venous measurements were undertaken in 3 animals. The PtO2 was 29 +/- 10 at an arterial pO2 of 116 +/- 10 (mean +/- SE, n = 5). The PvO2 was consistently higher than PtO2 although PvO2 approached PtO2 as PaO2 declined to 50 mmHg and was lower than tissue pO2 during the complete hypoxic period in one animal. These data indicate that brain venous pO2 is not representative of brain tissue pO2 and support published models predicting that in brain PvO2 is higher than PtO2.

Micro-imaging of articular cartilage: T2, proton density, and the magic angle effect.

RATIONALE AND OBJECTIVES: This study was designed to determine the relative influences of proton density versus collagen fiber orientation (through its influence on T2) in defining the layers of articular cartilage as seen in long-repetition-time magnetic resonance (MR) images. The authors mapped the T2 and proton densities of articular cartilage at 0 degree and 55 degrees with respect to the main magnetic field (B0) to determine the influence of T2 and water content on the normal laminar appearance of hyaline cartilage. MATERIALS AND METHODS: Six patellae of white-tailed deer were imaged at 7 T. T2 and proton densities were calculated from echo time versus signal intensity plots obtained with a multiecho, composite pulse sequence. Regions of interest in the radial and transitional zones were compared with the articular facets at 0 degree and 55 degrees relative to B0. Transmission electron microscopy was performed for correlation. RESULTS: At 0 degree, T2 was longer in the transitional than in the radial zone (29 vs 11 msec). AT 55 degrees, T2 increased in both radial and transitional zones, although the difference between the zones decreased (37 vs 29 msec). There was no difference in proton density between the two layers. CONCLUSION: Collagen fiber orientation, through T2 effects, is the dominant influence on the appearance of layers in hyaline cartilage in long-repetition-time MR images; proton density is not a major factor, and the collagen fiber orientation in the transitional zone is not totally random.

In vivo gradient echo microimaging of rodent spinal cord at 7 T.

An optimization scheme was developed for gradient echo imaging using a half-birdcage RF coil at 7 T to obtain maximal contrast between gray and white matter in the spinal cord of rodents. This optimization was combined with microimaging techniques to obtain in vivo pixel sizes of 78 x 78 x 700 microm. These techniques can be implemented in an in vivo study to investigate the myelin structure within the white matter of the rodent spinal cord.

Family structure and depressive symptoms in men preceding and following the birth of a child. The Avon Longitudinal Study of Pregnancy and Childhood Study Team.

OBJECTIVE: The prevalence and etiology of pre- and postpartum depressive symptoms in women in a variety of family forms have been well documented, but relatively little research has been conducted on the adjustment of their male partners. The authors' goals in this study were 1) to estimate rates of depression during the pregnancy and 8 weeks following the birth of a child in a large representative community sample of fathers in different family structures and 2) to explore the role of stressful life events, social and emotional support, the quality of the partner relationship, and socioeconomic circumstances. METHOD: This study describes the relations of family setting and other correlates to men's depressive symptoms during the pregnancies (18 weeks gestation, on average) and 8 weeks after the births of children for 7,018 partners of female participants in the Avon Longitudinal Study of Pregnancy and Childhood. RESULTS: Men living in stepfamilies had-significantly higher levels of depressive symptoms before and after the birth than did men in more traditional families. The effect of stepfamily status on depression was mediated by education, life events, social support, social network, and level of aggression in the partnership. CONCLUSIONS: There are similarities in the patterns and correlates of depression after the birth of a child for men and women. These findings point to the importance of family and partnership ecology in the adjustment of men before and after the birth of a child.