Acceptors stability modulates the efficiency of post-translational protein N-glycosylation.

N-glycosylation is the most common protein modification in the eukaryotic secretory pathway. It involves the attachment a high mannose glycan to Asn residues in the context of Asn-X-Ser/Thr/Cys, a motif known as N-glycosylation sequon. This process is mediated by STT3A and STT3B, the catalytic subunits of the oligosaccharyltransferase complexes. STT3A forms part of complexes associated with the SEC61 translocon and functions co-translationally. Vacant sequons have another opportunity for glycosylation by complexes carrying STT3B. Local sequence information plays an important role in determining N-glycosylation efficiency, but non-local factors can also have a significant impact. For instance, certain proteins associated with human genetic diseases exhibit abnormal N-glycosylation levels despite having wild-type acceptor sites. Here, we investigated the effect of protein stability on this process. To this end, we generated a family of 40 N-glycan acceptors based on superfolder GFP, and we measured their efficiency in HEK293 cells and in two derived cell lines lacking STT3B or STT3A. Sequon occupancy was highly dependent on protein stability, improving as the thermodynamic stability of the acceptor proteins decreases. This effect is mainly due to the activity of the STT3B-based OST complex. These findings can be integrated into a simple kinetic model that distinguishes local information within sequons from global information of the acceptor proteins.

Fragmented QRS complex could predict all-cause mortality in patients with connective tissue disease-associated pulmonary arterial hypertension.

OBJECTIVES: To investigate the prognostic impact and pathophysiological characteristics of fragmented QRS complex (fQRS) on patients with connective tissue disease-associated pulmonary arterial hypertension (CTD-PAH). METHODS: This was a multicentre retrospective study recruiting 141 patients with CTD-PAH diagnosed by right heart catheterization (114 cases in the discovery cohort and 27 cases in the validation cohort). fQRS and ST-T change were detected on conventional 12-lead electrocardiogram (ECG). Patients were followed up every 3 months to update their status and the primary end point was all-cause death. Clinical information and ECG characteristics were compared between survival and death groups and Kaplan-Meier curve was used for survival analysis. RESULTS: There were significant differences in age, gender, 6-min walk distance, NT-proBNP, WHO class, presence of fQRS and presence of ST-T change in inferior leads between survival group and death group. Inferior fQRS and ST-T change were significantly associated with right ventricular (RV) dilatation and reduced RV ejection fraction (RVEF). Kaplan-Meier curve showed that all-cause mortality was higher in CTD-PAH with fQRS (p= 0.003) and inferior ST-T change (p= 0.012). Low- and intermediate-risk CTD-PAH with inferior ST-T change had higher all-cause mortality (p= 0.005). The prognostic value of fQRS and inferior ST-T change was validated in external validation cohort. CONCLUSION: The presence of inferior fQRS and ST-T change could predict poor prognosis in CTD-PAH. CLINICAL TRIAL REGISTRATION NUMBER: NCT05980728, https://clinicaltrials.gov.

ST segment and T wave abnormalities: A narrative review.

INTRODUCTION: The electrocardiogram (ECG) is a valuable tool for interpreting ventricular repolarization. This article aims to broaden the diagnostic scope beyond the conventional ischemia-centric approach, integrating an understanding of pathophisiological influences on ST-T wave changes. METHODS: A review was conducted on the physiological underpinnings of ventricular repolarization and the pathophisiological processes that can change ECG patterns. The research encompassed primary repolarization abnormalities due to uniform variations in ventricular action potential, secondary changes from electrical or mechanical alterations, and non-ischemic conditions influencing ST-T segments. RESULTS: Primary T waves are characterized by symmetrical waves with broad bases and variable QT intervals, indicative of direct myocardial action potential modifications due to ischemia, electrolyte imbalances, and channelopathies. Secondary T waves are asymmetric and often unassociated with significant QT interval changes, suggesting depolarization alterations or changes in cardiac geometry and contractility. CONCLUSION: We advocate for a unified ECG analysis, recognizing primary and secondary ST-T changes, and their clinical implications. Our proposed analytical framework enhances the clinician's ability to discern a wide array of cardiac conditions, extending diagnostic accuracy beyond myocardial ischemia.

Ophthalmic findings associated with Australian tick paralysis (holocyclotoxicity) in hospitalized domestic dogs and cats.

OBJECTIVE: To describe ophthalmic findings in hospitalized canine and feline patients with tick paralysis (TP) and investigate possible predisposing factors. ANIMALS STUDIED: Forty-seven dogs and 28 cats hospitalized with TP assessed with an ophthalmic examination performed by an ABVO resident. METHODS: Dogs and cats were hospitalized with TP from October 2021 to January 2022 and had an ophthalmic examination performed by an ABVO resident. Patient signalment data, information regarding tick number and location, hospitalization duration, medications used, and patient paralysis grades were recorded. Statistical analysis was performed to correlate findings. RESULTS: Corneal ulcers developed in up to 34.8% of dogs and up to 42.9% of cats hospitalized with TP. An absent palpebral reflex ipsilaterally increased the odds of a concurrent corneal ulcer being present by 14.7× in dogs and 20.1× in cats (p < .0001). Palpebral reflexes were absent in 38.3% of dogs and 35.7% of cats hospitalized with TP and were correlated with more severe gait paralysis (p = .01) and respiratory paralysis (p = .005) in dogs, and respiratory paralysis in cats (p = .041). STT-1 findings <10 mm/min were present in 27.7% of dogs and 57.1% of cats examined and were associated with increasing gait paralysis (p = .017) and respiratory paralysis (p = .007) in dogs, and increasing gait paralysis in cats (p = .017). CONCLUSIONS: Simple corneal ulcers, loss of a complete palpebral reflex, and reduced STT-1 scores frequently occurred in dogs and cats hospitalized for TP. The frequency of these findings increased as the degree of patient paralysis increased.

Short-Term vs. Long-Term: A Critical Review of Indoor Radon Measurement Techniques.

Radon is a known carcinogen, and the accurate assessment of indoor levels is essential for effective mitigation strategies. While long-term testing provides the most reliable data, short-term testing (STT) offers a quicker and more cost-effective alternative. This review evaluated the accuracy of STT in predicting annual radon averages and compared testing strategies in Europe (where long-term measurements are common) and the United States (where STT is prevalent). Twenty (20) studies were systematically identified through searches in scientific databases and the grey literature, focusing on STT accuracy and radon management. This review revealed several factors that influence the accuracy of STT. Most studies recommended a minimum four-day test for initial screening, but accuracy varied with radon levels. For low levels (<75 Bq/m3), a one-week STT achieved high confidence (>95%) in predicting annual averages. However, accuracy decreased for moderate levels (approximately 50% success rate), necessitating confirmation with longer testing periods (3 months). High radon levels made STT unsuitable due to significant fluctuations. Seasonality also played a role, with winter months providing a more representative picture of annual radon averages. STT was found to be a useful method for screening low-risk areas with low radon concentrations. However, its limitations were evident in moderate- and high-level scenarios. While a minimum of four days was recommended, longer testing periods (3 months or more) were crucial for achieving reliable results, particularly in areas with potential for elevated radon exposure. This review suggests the need for further research to explore the possibility of harmonizing radon testing protocols between Europe and the United States.

Biomechanical evaluation of the wrist after scaphotrapeziotrapezoid arthrodesis.

BACKGROUND: The main objective of the present study was to present the biomechanical properties of the wrist in patients who underwent scaphotrapeziotrapezoid arthrodesis when compared to a healthy control hand. METHODS: The study group consisted of 29 consecutive patients who underwent a scaphotrapeziotrapezoid wrist arthrodesis at least 1 year before the research onset. Both hands of all patients were examined in 4 main categories. RESULTS: Average score obtained in the VAS, before the surgery, without motion of the wrist was 5.21 (SD = 3.04), whereas with wrist motion, it was 8.10 (SD = 1.37). Nineteen (65.52%) patients exhibited weakened wrist muscle strength. After the surgery, only 5 (17.24%) patients declared pain in the wrist. Furthermore, the results obtained in the VAS statistically significant differed from the ones before (p < 0.05). Twenty-eight (96.55%) patients were able to resume their profession. Twenty-seven (93.10%) patients stated that they would opt for the operation again. The peak torque during the analysis of extension of the wrist in the isometric protocol was found to be 8.1 Nm (SD = 2.9), 7.9 Nm (SD = 2.3), and 7.9 Nm (SD = 2.5) in the operated hands and 10.9 Nm (SD = 3.2), 9.6 Nm (SD = 2.9), and 9.1 Nm (SD = 3.8) in non-operated hand for 30° extension, no-flexion, and 30° flexion positions, respectively (p < 0.05). CONCLUSION: The current study is the first to present the biomechanical parameters of flexor and extensor muscles of the wrist and fingers in patients after the said procedure. Biomechanical assessments with additional isometric, isotonic, and isokinetic tests provide an opportunity to objectify treatment outcomes and guide appropriate rehabilitation by monitoring its effects. LEVEL OF EVIDENCE: III.

A cytosolic reductase pathway is required for efficient N-glycosylation of an STT3B-dependent acceptor site.

N-linked glycosylation of proteins entering the secretory pathway is an essential modification required for protein stability and function. Previously, it has been shown that there is a temporal relationship between protein folding and glycosylation, which influences the occupancy of specific glycosylation sites. Here, we used an in vitro translation system that reproduces the initial stages of secretory protein translocation, folding and glycosylation under defined redox conditions. We found that the efficiency of glycosylation of hemopexin was dependent upon a robust NADPH-dependent cytosolic reductive pathway, which could be mimicked by the addition of a membrane-impermeable reducing agent. We identified a hypoglycosylated acceptor site that is adjacent to a cysteine involved in a short-range disulfide. We show that efficient glycosylation at this site is influenced by the cytosolic reductive pathway acting on both STT3A- and STT3B-dependent glycosylation. Our results provide further insight into the important role of the endoplasmic reticulum redox conditions in glycosylation site occupancy and demonstrate a link between redox conditions in the cytosol and glycosylation efficiency.

Do facial soft tissue thicknesses change after surgeries correcting dental malocclusions? An intra- and inter-patient statistical analysis on soft-tissue thicknesses in BSSO + LFI surgeries.

OBJECTIVES: The aim of this study was to analyse changes in facial soft tissue thickness (FSTT) after corrective surgeries for dental malocclusion. The correlation between body mass index (BMI) and sex of patients and their FSTT before undergoing surgery was analysed. MATERIALS AND METHODS: Cone beam computed tomography of seventeen patients that underwent Le Fort I osteotomy in combination with bilateral sagittal split osteotomy were collected. Hard and soft tissue landmarks were selected basing on the interventions. FSTT were computed, and measurements from pre- to post-operative were compared. The relationship between FSTT, sex, and BMI was investigated. RESULTS: Considering the comparison between pre- and post-operative measurements, any significant difference emerged (p > .05). The Pearson's correlation coefficient computed between BMI and the FSTT (pre-operative) showed a correlation in normal-weight patients; the region-specific analysis highlighted a stronger correlation for specific landmarks. Higher median values emerged for women than for men; the subset-based analysis showed that women presented higher values in the malar region, while men presented higher values in the nasal region. CONCLUSIONS: The considered surgeries did not affect the FSTT of the patients; differences related to BMI and sex were found. A collection of FSTT mean values was provided for twenty landmarks of pre- and post-operative of female and male subjects. CLINICAL RELEVANCE: This exploratory analysis gave insights on the behaviour of STT after maxillofacial surgeries that can be applied in the development of predictive methodologies for soft tissue displacements and to study modifications in the facial aspect of the patients.

Tear film breakup time and Schirmer tear test in normal dogs: Effects of age, sex, reproductive status, skull type, and nasolacrimal duct patency.

OBJECTIVE: The present study aimed to determine the effects of age, sex, reproductive status, skull type, and nasolacrimal duct (NLD) patency on tear production and tear film breakup time (TBUT) in normal dogs. ANIMALS STUDIED: The ophthalmic data of 82 healthy adult dogs were evaluated in this study. PROCEDURES: Age, sex, breed, and reproductive status were recorded. Schirmer tear test (STT) and TBUT were assessed in all dogs, and interferometry was available for the selected dogs. The Jones test was used to evaluate NLD patency. The cephalic index (CI) was calculated for each dog (skull width/skull length ×100). RESULTS: Mean (SD) values for the STT results for the right (OD) and left (OS) eyes were 20.6 (2.7) and 20.2 (2.7) mm/min, respectively. Mean (SD) TBUT values for OD and OS were 6.5 (2.5) and 6.1 (2.3) mm/min in all dogs, respectively. Sex and reproductive status had no significant effect on STT and TBUT (P[OU] > 0.05). Skull type significantly affected TBUT in both eyes (P(OD)  = 0.01, P(OS)  = 0.003), but had no effect on STT (P[OU] > 0.3). Age had no correlation with STT and TBUT in either eye (P[OU] > 0.05). STT and TBUT had no correlation in either eye (P[OU] > 0.2). NLD patency had no significant effect on STT or TBUT (P[OU] > 0.1). CONCLUSIONS: The results of this study showed lower TBUT values in brachycephalic breeds than in non-brachycephalic breeds. A compensatory increase in STT values was observed in dogs with low TBUT values.

Quantitative Visualization of Thermally Enhanced Perpendicular Shape Anisotropy STT-MRAM Nanopillars.

Perpendicular shape anisotropy (PSA) offers a practical solution to downscale spin-transfer torque magnetoresistive random-access memory (STT-MRAM) beyond the sub-20 nm technology node while retaining thermal stability. However, our understanding of the thermomagnetic behavior of PSA-STT-MRAM is often indirect, relying on magnetoresistance measurements and micromagnetic modeling. Here, the magnetism of a NiFe PSA-STT-MRAM nanopillar is investigated using off-axis electron holography, providing spatially resolved magnetic information as a function of temperature. Magnetic induction maps reveal the micromagnetic configuration of the NiFe storage layer (∼60 nm high, ≤20 nm diameter), confirming the PSA induced by its 3:1 aspect ratio. In situ heating demonstrates that the PSA of the storage layer is maintained up to at least 250 °C, and direct quantitative measurements reveal a moderate decrease of magnetic induction. Hence, this study shows explicitly that PSA provides significant stability in STT-MRAM applications that require reliable performance over a range of operating temperatures.

Overexpression of TaSTT3b-2B improves resistance to sharp eyespot and increases grain weight in wheat.

STAUROSPORINE AND TEMPERATURE SENSITIVE3 (STT3) is a catalytic subunit of oligosaccharyltransferase, which is important for asparagine-linked glycosylation. Sharp eyespot, caused by the necrotrophic fungal pathogen Rhizoctonia cerealis, is a devastating disease of bread wheat. However, the molecular mechanisms underlying wheat defense against R. cerealis are still largely unclear. In this study, we identified TaSTT3a and TaSTT3b, two STT3 subunit genes from wheat and reported their functional roles in wheat defense against R. cerealis and increasing grain weight. The transcript abundance of TaSTT3b-2B was associated with the degree of wheat resistance to R. cerealis and induced by both R. cerealis and exogenous jasmonic acid (JA). Overexpression of TaSTT3b-2B significantly enhanced resistance to R. cerealis, grain weight, and JA content in transgenic wheat subjected to R. cerealis stress, while silencing of TaSTT3b-2B compromised resistance of wheat to R. cerealis. Transcriptomic analysis showed that TaSTT3b-2B affected the expression of a series of defense-related genes and JA biosynthesis-related genes, as well as genes coding starch synthase and sucrose synthase. Application of exogenous JA elevated expression levels of the abovementioned defense- and grain weight-related genes, and rescuing the resistance of TaSTT3b-2B-silenced wheat to R. cerealis, while pretreatment with sodium diethyldithiocarbamate, an inhibitor of JA synthesis, attenuated the TaSTT3b-2B-mediated resistance to R. cerealis, suggesting that TaSTT3b-2B played critical roles in regulating R. cerealis resistance and grain weight via JA biosynthesis. Altogether, this study reveals new functional roles of TaSTT3b-2B in regulating plant innate immunity and grain weight, and illustrates its potential application value for wheat molecular breeding.

Elevation of spermine remodels immunosuppressive microenvironment through driving the modification of PD-L1 in hepatocellular carcinoma.

BACKGROUND: Spermine is frequently elevated in tumor tissues and body fluids of cancer patients and is critical for cancer cell proliferation, migration and invasion. However, the immune functions of spermine in hepatocellular carcinoma progression remains unknown. In the present study, we aimed to elucidate immunosuppressive role of spermine in hepatocellular carcinoma and to explore the underlying mechanism. METHODS: Whole-blood spermine concentration was measured using HPLC. Human primary HCC tissues were collected to examine the expression of CaSR, p-Akt, ß-catenin, STT3A, PD-L1, and CD8. Mouse model of tumorigenesis and lung metastasis were established to evaluate the effects of spermine on hepatocellular carcinoma. Western blotting, immunofluorescence, real time PCR, digital Ca2+ imaging, and chromatin immunoprecipitation assay were used to investigate the underlying mechanisms by which spermine regulates PD-L1 expression and glycosylation in hepatocellular carcinoma cells. RESULTS: Blood spermine concentration in the HCC patient group was significantly higher than that in the normal population group. Spermine could facilitate tumor progression through inducing PD-L1 expression and decreasing the CD8+ T cell infiltration in HCC. Mechanistically, spermine activates calcium-sensing receptor (CaSR) to trigger Ca2+ entry and thereby promote Akt-dependent ß-catenin stabilization and nuclear translocation. Nuclear ß-catenin induced by spermine then activates transcriptional expression of PD-L1 and N-glycosyltransferase STT3A, while STT3A in turn increases the stability of PD-L1 through inducing PD-L1 protein N-glycosylation in HCC cells. CONCLUSIONS: This study reveals the crucial function of spermine in establishing immune privilege by increasing the expression and N-glycosylation of PD-L1, providing a potential strategy for the treatment of hepatocellular carcinoma. Video Abstract.

Ligaments of the scapho-trapezial-trapezoidal joint: MR anatomy in asymptomatic and symptomatic individuals.

PURPOSE: To evaluate the MRI anatomy of the scapho-trapezial-trapezoidal (STT) ligament complex in asymptomatic and symptomatic individuals. MATERIAL AND METHODS: In this retrospective study, STT ligament complex of 42 (male 69%, median age 37.5 years) asymptomatic (n = 25) and symptomatic (n = 17) (defined as pain described over the STT joint) individuals was examined using a high-resolution 3D proton density-weighted isovoxel sequence (MR arthrogram) with multiplanar reconstructions. Two musculoskeletal radiologists independently assessed visibility, signal intensity (SI), morphology, and thickness of the radiopalmar scapho-trapezial ligament (rpSTL), palmar scapho-capitate capsular ligament (pSCL), palmar STT capsule (pSTTC), and dorsal STT capsule (dSTTC). RESULTS: Interreader agreement ranged from fair to good and intraclass correlations were good. The rpSTL was almost always visible (85.7%/80.1%; reader 1/reader 2). The pSCL and dSTTC were visible in all cases. The pSTTC was visible in only 52.4%/42.9%. Mean thickness of the rpSTL, pSCL, pSTTC, and dSTTC was 1.4 ± 0.5 mm/1.3 ± 0.5 mm, 2.8 ± 0.7 mm/2.7 ± 0.6 mm, 0.5 ± 0.5 mm/0.4 ± 0.4 mm, and 0.5 ± 0.3 mm/0.3 ± 0.3 mm. Both readers rated SI of the rpSTL significantly more often as increased in the symptomatic group (increased SI in asymptomatic group: 20%/15%; symptomatic group: 56%/50%) (p-values < 0.005). For all other ligaments, no significant difference was observed for SI between symptomatic and asymptomatic group (p-values ranging between 0.188 and 0.890). For all other ligaments, no significant differences were observed regarding ligament visibility, morphology, and thickness (p-values ranging between 0.274 and 1.000). CONCLUSION: The anatomy of the STT ligament complex can consistently be visualized on high-resolution 3D MRI. Increased signal intensity of rpSTL is significantly more frequent in patients with radial-sided wrist pain.

Posterior tibial tubercle measured by the sagittal TT-TG distance correlates with increased risk for patellofemoral chondral lesions.

PURPOSE: To evaluate the variation in tibial tubercle sagittal alignment in patients with and without patellofemoral (PF) cartilage wear. METHODS: This was a single-centre, retrospective review of patients that underwent a cartilage restoration procedure for isolated PF cartilage wear from 2014 to 2020. Patients were matched in a 1:2 ratio for age, sex and BMI to partial meniscectomy patients as controls. The sagittal TT-TG (sTT-TG) distance was measured on preoperative axial T2 magnetic resonance imaging (MRI) and was defined as the distance between a point at the nadir of the trochlear cartilage and the most anterior point of the tibial tubercle. RESULTS: One hundred and forty patients (47 cartilage restoration, 94 meniscectomy) were included. Mean age, BMI, and height for the total cohort were 34.01 ± 8.7, 26.6 ± 6.4, and 173.0 ± 17.7 respectively, with 78 males (55%) and 63 females (45%). There were no significant differences between groups for age, BMI or sex (n.s). The cartilage restoration group (- 2.5 mm ± 5.9) was found to have a significantly more posterior (negative) sTT-TG compared to the meniscectomy group (1.72 mm ± 6.7) (p < 0.001). Interrater reliability was excellent (ICC = 0.931, p < 0.001). Patients with less than - 3.4 mm sTT-TG were 2.74 times more likely to have a cartilage restoration procedure compared to those with greater than - 3.4 mm (OR 2.7, 95% CI 1.3-5.85). Patients with < - 10 mm posterior translation were 13.7× (CI 1.6-111.1) more likely to have a cartilage restoration procedure. CONCLUSION: Patients that underwent isolated cartilage restoration procedures had a significantly more posterior tibial tubercle than partial meniscectomy controls based on the sagittal TT-TG. The more posterior the tubercle, the more likely the patient had a cartilage restoration procedure. Surgeons should consider the sTT-TG measurement in patients presenting with anterior knee pain, particularly patellofemoral lesions. LEVEL OF EVIDENCE: III.

Scaphotrapeziotrapezoid Arthrodesis: Systematic Review.

PURPOSE: Scaphotrapeziotrapezoid (STT) arthrodesis surgery is used for various types of wrist pathologies. The objective of our study was to perform a systematic review of complications and outcomes after STT arthrodesis. METHODS: Several major databases were used to perform a systematic literature review in order to obtain articles reporting complications and outcomes following STT arthrodesis. The primary purpose was to identify rates of nonunion and conversion to total wrist arthrodesis. Secondary outcomes included wrist range of motion, grip strength, and Disabilities of the Arm Shoulder and Hand scores. A multivariable analysis was performed to evaluate factors associated with the primary and secondary outcomes of interest. RESULTS: Out of the 854 records identified in the primary literature search, 30 studies were included in the analysis. A total of 1,429 procedures were performed for 1,404 patients. The pooled nonunion rate was 6.3% (95% CI, 3.5-9.9) and the rate of conversion to total wrist arthrodesis following the index STT was 4.2% (95% CI, 2.2-6.7). The mean pooled wrist flexion was 40.7° (95% CI, 30.8-50.5) and extension was 49.7° (95% CI, 43.5-55.8). At final follow-up, the mean pooled grip strength was 75.9% (95% CI, 69.3-82.5) of the nonsurgical contralateral hand. Compared with all other known indications, Kienbock disease had a statistically significant lower nonunion rate (14.1% vs 3.3%, respectively). Mixed-effects linear regression using patient-level data revealed that increasing age was significantly associated with complications, independent of occupation and diagnosis. CONCLUSIONS: Our study demonstrated a low failure rate and conversion to total wrist arthrodesis after STT arthrodesis and acceptable postoperative wrist range of motion and strength when compared to the contralateral hand. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV.

Electrocardiographic Changes in COVID-19 Patients: A Hospital-based Descriptive Study.

BACKGROUND: Coronavirus disease-2019 (COVID-19) infection is a multisystem disease not restricted to the lungs. It has a negative impact on the cardiovascular system by causing myocardial damage, vascular inflammation, plaque instability, and myocardial infarction. The presence of myocardial injury is a poor prognostic sign. Electrocardiogram (ECG), a simple bedside diagnostic test with high prognostic value, can be employed to assess early cardiovascular involvement in such patients. Various abnormalities in ECG like ST-T changes, arrhythmia, and conduction defects have been reported in COVID-19. We aimed to find out the ECG abnormalities of COVID-19 patients. METHODS: We performed a cross-sectional, hospital-based descriptive study among 315 COVID-19 in-patients who underwent ECG recording on admission. Patients' clinical profiles were noted from their records, and the ECG abnormalities were studied. RESULTS: Among the abnormal ECGs 255 (81%), rhythm abnormalities were seen in 9 patients (2.9%), rate abnormalities in 115 patients (36.5%), and prolonged PR interval in 2.9%. Short QRS complex was seen in 8.3%. QT interval was prolonged in 8.3% of the patients. Significant changes in the ST and T segments (42.9%) were observed. In logistic regression analysis, ischemic changes in ECG were associated with systemic hypertension and respiratory failure. CONCLUSION: In our study, COVID-19 patients had ischemic changes, rate, rhythm abnormalities, and conduction defects in their ECG. With this ongoing pandemic of COVID-19 and limited health resources, ECG-a simple bedside noninvasive tool is highly beneficial and helps in the early diagnosis and management of cardiac injury. HOW TO CITE THIS ARTICLE: Kaliyaperumal D, Bhargavi K, Ramaraju K, Nair KS, Ramalingam S, Alagesan M. Electrocardiographic Changes in COVID-19 Patients: A Hospital-based Descriptive Study. Indian J Crit Care Med 2022;26(1):43-48.

Mammalian STT3A/B oligosaccharyltransferases segregate N-glycosylation at the translocon from lipid-linked oligosaccharide hydrolysis.

Oligosaccharyltransferases (OSTs) N-glycosylate proteins by transferring oligosaccharides from lipid-linked oligosaccharides (LLOs) to asparaginyl residues of Asn-Xaa-Ser/Thr acceptor sequons. Mammals have OST isoforms with STT3A or STT3B catalytic subunits for cotranslational or posttranslational N-glycosylation, respectively. OSTs also hydrolyze LLOs, forming free oligosaccharides (fOSs). It has been unclear whether hydrolysis is due to one or both OSTs, segregated from N-glycosylation, and/or regulated. Transfer and hydrolysis were assayed in permeabilized HEK293 kidney and Huh7.5.1 liver cells lacking STT3A or STT3B. Transfer by both STT3A-OST and STT3B-OST with synthetic acceptors was robust. LLO hydrolysis by STT3B-OST was readily detected and surprisingly modulated: Without acceptors, STT3B-OST hydrolyzed Glc3Man9GlcNAc2-LLO but not Man9GlcNAc2-LLO, yet it hydrolyzed both LLOs with acceptors present. In contrast, LLO hydrolysis by STT3A-OST was negligible. STT3A-OST however may be regulatory, because it suppressed STT3B-OST-dependent fOSs. TREX1, a negative innate immunity factor that diminishes immunogenic fOSs derived from LLOs, acted through STT3B-OST as well. In summary, only STT3B-OST hydrolyzes LLOs, depending upon LLO quality and acceptor site occupancy. TREX1 and STT3A suppress STT3B-OST-dependent fOSs. Without strict kinetic limitations during posttranslational N-glycosylation, STT3B-OST can thus moonlight for LLO hydrolysis. In contrast, the STT3A-OST/translocon complex preserves LLOs for temporally fastidious cotranslational N-glycosylation.

Comprehensive Interactome Analysis Reveals that STT3B Is Required for N-Glycosylation of Lassa Virus Glycoprotein.

Lassa virus (LASV) is the causative agent of a fatal hemorrhagic fever in humans. The glycoprotein (GP) of LASV mediates viral entry into host cells, and correct processing and modification of GP by host factors is a prerequisite for virus replication. Here, using an affinity purification-coupled mass spectrometry (AP-MS) strategy, 591 host proteins were identified as interactors of LASV GP. Gene ontology analysis was performed to functionally annotate these proteins, and the oligosaccharyltransferase (OST) complex was highly enriched. Functional studies conducted by using CRISPR-Cas9-mediated knockouts showed that STT3A and STT3B, the two catalytically active isoforms of the OST complex, are essential for the propagation of the recombinant arenavirus rLCMV/LASV glycoprotein precursor, mainly via affecting virus infectivity. Knockout of STT3B, but not STT3A, caused hypoglycosylation of LASV GP, indicating a preferential requirement of LASV for the STT3B-OST isoform. Furthermore, double knockout of magnesium transporter 1 (MAGT1) and tumor suppressor candidate 3 (TUSC3), two specific subunits of STT3B-OST, also caused hypoglycosylation of LASV GP and affected virus propagation. Site-directed mutagenesis analysis revealed that the oxidoreductase CXXC active-site motif of MAGT1 or TUSC3 is essential for the glycosylation of LASV GP. NGI-1, a small-molecule OST inhibitor, can effectively reduce virus infectivity without affecting cell viability. The STT3B-dependent N-glycosylation of GP is conserved among other arenaviruses, including both the Old World and New World groups. Our study provided a systematic view of LASV GP-host interactions and revealed the preferential requirement of STT3B for LASV GP N-glycosylation.IMPORTANCE Glycoproteins play vital roles in the arenavirus life cycle by facilitating virus entry and participating in the virus budding process. N-glycosylation of GPs is responsible for their proper functioning; however, little is known about the host factors on which the virus depends for this process. In this study, a comprehensive LASV GP interactome was characterized, and further study revealed that STT3B-dependent N-glycosylation was preferentially required by arenavirus GPs and critical for virus infectivity. The two specific thioredoxin subunits of STT3B-OST MAGT1 and TUSC3 were found to be essential for the N-glycosylation of viral GP. NGI-1, a small-molecule inhibitor of OST, also showed a robust inhibitory effect on arenavirus. Our study provides new insights into LASV GP-host interactions and extends the potential targets for the development of novel therapeutics against Lassa fever in the future.

Inflammatory conditions promote a switch of oligosaccharyltransferase (OST) catalytic subunit isoform expression.

Oligosaccharyltransferase (OST) complex catalyzes the N-glycosylation of nascent polypeptides in the endoplasmic reticulum. Glycoproteins are critical for normal cell-cell interactions, especially during an immune response. Abnormal glycosylation is an insignia of several inflammatory diseases. However, the mechanisms that regulate the differential N-glycosylation are not fully understood. The OST complex can be assembled with one out of two catalytic subunits, STT3A or STT3B, which have different enzymatic properties. In this work, we investigated the expression of STT3A and STT3B in several mouse models such as a crossbreeding of normal and abortion-prone mice and an intestinal inflammation model. These animals were either exposed or not to acoustic stress (acute or chronic). The expression of the isoforms was analysed by immunohistochemistry and protein immunoblot. Finally, we investigated the gene regulatory elements employing public databases. Results demonstrated that inflammation alters the balance between the expression of both isoforms in the affected tissues. In homoeostatic conditions, STT3A expression predominates over STT3B, especially in epithelial cells. This relation is reversed as a consequence of inflammation. An increase in STT3B activity was associated to the generation of mannose-rich N-glycans. Accordingly, this type of N-glycans were found to decorate diverse inflamed tissues. The STT3A and STT3B genes are differentially regulated, which could account for the differences in the expression levels observed here. Our results support the idea that these isoforms could play different roles in cellular physiology. This study opens the possibility of studying the STT3A/STT3B expression ratio as a biomarker in acute inflammation or chronic diseases.

Ascorbate inducible N259 glycans on prolyl 4-hydroxylase subunit α1 promote hydroxylation and secretion of type I collagen.

Ascorbic acid (vitamin C, VC) increases the secretion of mature collagen by promoting the activity of prolyl 4-hydroxylase subunit α 1 (P4HA1). To explore the mechanism involved, we investigated the role of N-linked glycosylation, which can regulate enzyme activity. P4HA1 has two glycosylation sites, Asn (N) 113 and N259. Our studies show that glycosylation of N259, but not N113, by STT3B and magnesium transporter 1 (MAGT1) is augmented by VC. N259 glycosylation on P4HA1 correlates with enhanced pepsin-resistant collagen 1α2 secretion. Downregulation of Stt3b and Magt1 reduces N259 glycans on P4HA1. In collagen 1α2 purified from Stt3b-silenced fibroblasts, decreased hydroxylation is found at five specific proline residues, while significantly increased hydroxylation is noted at two proline residues. Similarly, in collagen 1α1, reduced proline hydroxylation is detected at eight sites and increased proline hydroxylation is found at four sites. These results suggest that N-linked glycosylation of P4HA1 can direct hydroxylation at specific proline residues and affect collagen maturation.