Oxidation products of DNA, lipid and protein among the individuals progressing towards metabolic syndrome

Oxidative stress (OS) is an early event and at the same time also a consequence in the pathology of MetS. We investigated if oxidation markers of DNA, lipid and protein increased with an increase in the risk parameters of MetS. Participants (male:70, female:90 ≥ 20 yrs) were categorized based on the number of risk factors they had as 3 Risk, 2 Risk, 1 Risk and 0 Risk for MetS and were evaluated for various oxidation markers. Protein carbonyl and advanced oxidation protein product (protein oxidation marker) differed significantly between the four study group while malondialdehyde and hydroxynonenal (lipid peroxidation marker) did not. “8-OH dG” (DNA oxidation marker) differed significantly (P< 0.05) while total antioxidant capacity did not demonstrate significant difference in its values across the group (P> 0.05). Pairwise comparison for statistically significant markers(Protein oxidation markers and 8-OH dG), demonstrated that only 8-OH dG differed significantly between 0 Risk- 3 Risk (P< 0.012) but not between 0 Risk -2 Risk and 0 Risk-1 Risk. Oxidative stress markers of DNA, lipid and protein do not increase with an increase in the risk parameters of MetS. However, it is indeed high in MetS with 3 and more risk parameters. Presence of 2 or 1 Risk also increases OS compared to 0 Risk. There is oxidative stress damage in MetS to lipid and protein but DNA damage was of significant consequence

Neurovascular Coupling Remains Intact During Incremental Ascent to High Altitude (4240 m) in Acclimatized Healthy Volunteers

Neurovascular coupling (NVC) is the temporal link between neuronal metabolic activity and regional cerebral blood flow (CBF), supporting adequate delivery of nutrients. Exposure to high altitude (HA) imposes several stressors, including hypoxia and hypocapnia, which modulate cerebrovascular tone in an antagonistic fashion. Whether these contrasting stressors and subsequent adaptations affect NVC during incremental ascent to HA is unclear. The aim of this study was to assess whether incremental ascent to HA influences the NVC response. Given that CBF is sensitive to changes in arterial blood gasses, in particular PaCO2, we hypothesized that the vasoconstrictive effect of hypocapnia during ascent would decrease the NVC response. 10 healthy study participants (21.7 ± 1.3 years, 23.57 ± 2.00 kg/m2, mean ± SD) were recruited as part of a research expedition to HA in the Nepal Himalaya. Resting posterior cerebral artery velocity (PCAv), arterial blood gasses (PaO2, SaO2, PaCO2, [HCO3-], base excess and arterial blood pH) and NVC response of the PCA were measured at four pre-determined locations: Calgary/Kathmandu (1045/1400 m, control), Namche (3440 m), Deboche (3820 m) and Pheriche (4240 m). PCAv was measured using transcranial Doppler ultrasound. Arterial blood draws were taken from the radial artery and analyzed using a portable blood gas/electrolyte analyzer. NVC was determined in response to visual stimulation (VS; Strobe light; 6 Hz; 30 s on/off × 3 trials). The NVC response was averaged across three VS trials at each location. PaO2, SaO2, and PaCO2 were each significantly decreased at 3440, 3820, and 4240 m. No significant differences were found for pH at HA (P > 0.05) due to significant reductions in [HCO3-] (P < 0.043). As expected, incremental ascent to HA induced a state of hypoxic hypocapnia, whereas normal arterial pH was maintained due to renal compensation. NVC was quantified as the delta (Δ) PCAv from baseline for mean PCAv, peak PCAv and total area under the curve (ΔPCAv tAUC) during VS. No significant differences were found for Δmean, Δpeak or ΔPCAv tAUC between locations (P > 0.05). NVC remains remarkably intact during incremental ascent to HA in healthy acclimatized individuals. Despite the array of superimposed stressors associated with ascent to HA, CBF and NVC regulation may be preserved coincident with arterial pH maintenance during acclimatization

Cardiorespiratory hysteresis during incremental high altitude ascent-descent quantifies the magnitude of ventilatory acclimatization

Maintenance of arterial blood gases is achieved through sophisticated regulation of ventilation, mediated by central and peripheral chemoreflexes. Respiratory chemoreflexes are important during exposure to high altitude due to the competing influence of hypoxia and hypoxic hyperventilation‐mediated hypocapnia on steady‐state ventilatory drive. Inter‐individual variability exists in ventilatory acclimatization to high altitude, potentially affecting the development of acute mountain sickness (AMS). We aimed to quantify ventilatory acclimatization to high altitude by comparing differential ascent and descent values (i.e. hysteresis) in steady‐state cardiorespiratory variables. We hypothesized that (a) the hysteresis area formed by cardiorespiratory variables during ascent and descent would quantify the magnitude of ventilatory acclimatization, and (b) larger hysteresis areas would be associated with lower AMS symptom scores during ascent. In 25 healthy, Diamox‐free trekkers ascending to and descending from 5160 m, cardiorespiratory hysteresis was measured in the pressure of end‐tidal (PET)CO2, peripheral oxygen saturation (SpO2), minute ventilation (V̇E), chemoreceptor stimulus index (SI; PETCO2/SpO2) and the calculated steady‐state chemoreflex drive (SS‐CD; V̇E/SI) using portable devices (capnograph, peripheral pulse oximeter and respirometer, respectively). AMS symptoms were assessed daily using the Lake Louise Questionnaire. We found that (a) ascent‐descent hysteresis was present in all cardiorespiratory variables, (b) SS‐CD is a valid metric for tracking ventilatory acclimatization to high altitude and (c) highest AMS scores during ascent were significantly, moderately and inversely‐correlated to SS‐CD hysteresis magnitude (rs = ‐0.408, P = 0.043). We propose that ascent‐descent hysteresis is a novel and feasible way to quantify ventilatory acclimatization in trekkers during high altitude exposure

Biallelic Mutations in MRPS34 Lead to Instability of the Small Mitoribosomal Subunit and Leigh Syndrome

The synthesis of all 13 mitochondrial DNA (mtDNA)-encoded protein subunits of the human oxidative phosphorylation (OXPHOS) system is carried out by mitochondrial ribosomes (mitoribosomes). Defects in the stability of mitoribosomal proteins or mitoribosome assembly impair mitochondrial protein translation, causing combined OXPHOS enzyme deficiency and clinical disease. Here we report four autosomal-recessive pathogenic mutations in the gene encoding the small mitoribosomal subunit protein, MRPS34, in six subjects from four unrelated families with Leigh syndrome and combined OXPHOS defects. Whole-exome sequencing was used to independently identify all variants. Two splice-site mutations were identified, including homozygous c.321+1G>T in a subject of Italian ancestry and homozygous c.322−10G>A in affected sibling pairs from two unrelated families of Puerto Rican descent. In addition, compound heterozygous MRPS34 mutations were identified in a proband of French ancestry; a missense (c.37G>A [p.Glu13Lys]) and a nonsense (c.94C>T [p.Gln32∗]) variant. We demonstrated that these mutations reduce MRPS34 protein levels and the synthesis of OXPHOS subunits encoded by mtDNA. Examination of the mitoribosome profile and quantitative proteomics showed that the mitochondrial translation defect was caused by destabilization of the small mitoribosomal subunit and impaired monosome assembly. Lentiviral-mediated expression of wild-type MRPS34 rescued the defect in mitochondrial translation observed in skin fibroblasts from affected subjects, confirming the pathogenicity of MRPS34 mutations. Our data establish that MRPS34 is required for normal function of the mitoribosome in humans and furthermore demonstrate the power of quantitative proteomic analysis to identify signatures of defects in specific cellular pathways in fibroblasts from subjects with inherited disease

Oxidation products of DNA, lipid and protein among the individuals progressing towards metabolic syndrome

155-161Oxidative stress (OS) is an early event and at the same time also a consequence in the pathology of MetS. We investigated if oxidation markers of DNA, lipid and protein increased with an increase in the risk parameters of MetS. Participants (male:70, female:90 ≥ 20 yrs) were categorized based on the number of risk factors they had as 3 Risk, 2 Risk, 1 Risk and 0 Risk for MetS and were evaluated for various oxidation markers. Protein carbonyl and advanced oxidation protein product (protein oxidation marker) differed significantly between the four study group while malondialdehyde and hydroxynonenal (lipid peroxidation marker) did not. “8-OH dG” (DNA oxidation marker) differed significantly (PP> 0.05). Pairwise comparison for statistically significant markers(Protein oxidation markers and 8-OH dG), demonstrated that only 8-OH dG differed significantly between 0 Risk- 3 Risk (P< 0.012) but not between 0 Risk -2 Risk and 0 Risk-1 Risk. Oxidative stress markers of DNA, lipid and protein do not increase with an increase in the risk parameters of MetS. However, it is indeed high in MetS with 3 and more risk parameters. Presence of 2 or 1 Risk also increases OS compared to 0 Risk. There is oxidative stress damage in MetS to lipid and protein but DNA damage was of significant consequence. </span

Heterozygous Pathogenic Variant in DACT1 Causes an Autosomal-Dominant Syndrome with Features Overlapping Townes-Brocks Syndrome

A heterozygous nonsense variant was identified in dapper, antagonist of beta-catenin, 1 (DACT1) via whole-exome sequencing in family members with imperforate anus, structural renal abnormalities, genitourinary anomalies, and/or ear anomalies. The DACT1 c.1256G>A;p.Trp419* variant segregated appropriately in the family consistent with an autosomal dominant mode of inheritance. DACT1 is a member of the Wnt-signaling pathway, and mice homozygous for null alleles display multiple congenital anomalies including absent anus with blind-ending colon and genitourinary malformations. To investigate the DACT1 c.1256G>A variant, HEK293 cells were transfected with mutant DACT1 cDNA plasmid, and immunoblotting revealed stability of the DACT1 p.Trp419* protein. Overexpression of DACT1 c.1256G>A mRNA in Xenopus embryos revealed a specific gastrointestinal phenotype of enlargement of the proctodeum. Together, these findings suggest that the DACT1 c.1256G>A nonsense variant is causative of a specific genetic syndrome with features overlapping Townes-Brocks syndrom

Renal Acid-Base Compensation Demonstrates Plasticity During Incremental Ascent to High Altitude

Ascent to high altitude, and the associated hypoxic ventilatory response, imposes an acid-base challenge, namely chronic hypocapnia and respiratory alkalosis. The kidneys act to compensate for this respiratory alkalosis via bicarbonate (HCO3-) excretion in urine to induce a compensatory metabolic acidosis. The time course and extent of plasticity of this important renal response during incremental ascent to altitude is unclear. We developed a practical index of renal reactivity (RR), indexing the relative change in arterial HCO3- concentration ([HCO3-]a; i.e., response) against the relative change in arterial partial pressure of CO2 (PaCO2; i.e., stimulus) during ascent (i.e., RR=Δ[HCO3-]a/ΔPaCO2). We sought to assess if RR increased over time and with incremental ascent to altitude, and if RR was correlated with relative changes in arterial pH (ΔpHa) throughout ascent. During ascent to 5160m over 10 days in the Nepal Himalaya, arterial blood was drawn from the radial artery for measurement of acid-base variables (Abbott iSTAT portable blood gas/electrolyte analyzer; CG4+ and CHEM8+ cartridges) in lowlanders at 1045/1400m (baseline) and at four different altitudes following one-night sleep: 3440m, 3820m, 4370m and 5160m. At 3820m (day five) and higher, RR significantly increased and plateaued in comparison to 3440m (day three; P&lt;0.04), suggesting plasticity in renal acid-base compensation. At all four altitudes, we observed a strong correlation (range: r=-0.71 to -0.98; P&lt;0.001) between RR and relative ΔpHa from baseline, suggesting that the RR index accurately quantified renal acid-base responsiveness throughout ascent. In conclusion, renal acid-base compensation mechanisms demonstrate plasticity during incremental ascent to high altitude, which was detected using a novel RR index. The extent of plasticity and plateau in renal responsiveness may predict severity of altitude illness or acclimatization at higher or more prolonged stays at altitude. Support or Funding Information: This work was supported by (a) Alberta Government Student Temporary Employment Program, (b) Alberta Innovates Health Solutions Summer Studentship, and (c) Natural Sciences and Engineering Research Council of Canada Discovery grant. *Indicates presente

Galectin-3 Is Associated with Cardiac Fibrosis and an Increased Risk of Sudden Death

Background: Myocardial fibrosis is a common postmortem finding among individuals with Sudden Cardiac Death (SCD). Numerous in vivo and in vitro studies have shown that increased galectin-3 (gal3) expression into the myocardium is associated with higher incidence of fibrosis. Although elevated gal3 expression is linked with myocardial fibrosis, its role in predicting the risk of SCD is unknown. Methods: We reviewed the clinical datasets and post-mortem examination of 221 subjects who had died suddenly. We examined myocardial pathology including the extent of cardiac hypertrophy, fibrosis, and the degree of coronary atherosclerosis in these subjects. In a select group of SCD subjects, we studied myocardial gal3 and periostin expression using immunohistochemistry. To further examine if a higher level of circulating gal3 can be detected preceding sudden death, we measured serum gal3 in a porcine model of subtotal coronary artery ligation which shows an increased tendency to develop lethal cardiac arrhythmias, including ventricular tachycardia or fibrillation. Results: Of the total 1314 human subjects screened, 12.7% had SCD. Comparison of age-matched SCD with non-SCD subjects showed that SCD groups had excessive myocardial fibrosis involving both the left ventricular free wall and interventricular septum. In pigs with subtotal coronary artery ligation and SCD, we detected significantly elevated circulating gal3 levels approximately 10 days preceding the SCD event. Immunohistochemistry showed increased myocardial gal3 and periostin expression in pigs that died suddenly, compared to the controls. Conclusion: Our study shows that increased gal3 is associated with a higher risk of myocardial fibrosis and the risk of SCD. This supports the importance of larger translational studies to target gal3 to prevent cardiac fibrosis and attenuate the risk of SCD