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1.
J Clin Neuromuscul Dis ; 23(4): 189-200, 2022 Jun 01.
Article in English | MEDLINE | ID: covidwho-1860946

ABSTRACT

ABSTRACT: This update covers a number of treatment topics starting with Fc receptor inhibitors and the Federal Drug Administration approval of efgartigimod. Some uncertainties regarding the use of corticosteroids are addressed, namely the risk of exacerbation with initiation of treatment and how to taper. The presence and potential importance of antibody overshoot following plasmapheresis is noted and the evolving increase in usefulness of acetylcholine receptor antibodies in diagnosing ocular myasthenia. Several recent series and case reports regarding coronavirus 2019 and myasthenia gravis are reviewed. The topics of myasthenia gravis and pregnancy, and another look at thymectomy in MG are provided. Finally, a couple of case reports on Lambert-Eaton myasthenic syndrome concentrate on the ice pack test and an autoantibody association with paraneoplastic cerebellar degeneration and Lambert-Eaton myasthenic syndrome in the same patient.


Subject(s)
Lambert-Eaton Myasthenic Syndrome , Myasthenia Gravis , Autoantibodies , Humans , Lambert-Eaton Myasthenic Syndrome/diagnosis , Myasthenia Gravis/complications , Myasthenia Gravis/diagnosis , Myasthenia Gravis/therapy , Neuromuscular Junction , Receptors, Cholinergic
2.
Neurol Sci ; 43(7): 4081-4083, 2022 Jul.
Article in English | MEDLINE | ID: covidwho-1777738

ABSTRACT

Eculizumab, a humanized monoclonal antibody, is a complement inhibitor indicated for refractory generalized myasthenia gravis (MG). However, there are limited data on the safety of eculizumab for MG during coronavirus disease 2019 (COVID-19) infection. We report a case in which eculizumab was continued for MG after contracting COVID-19, followed by a favorable outcome.


Subject(s)
COVID-19 , Myasthenia Gravis , Antibodies, Monoclonal, Humanized/therapeutic use , Complement Inactivating Agents/therapeutic use , Humans , Myasthenia Gravis/complications , Myasthenia Gravis/drug therapy
3.
Acta Clin Croat ; 60(3): 496-509, 2022 Feb.
Article in English | MEDLINE | ID: covidwho-1727116

ABSTRACT

Coronavirus disease 2019 (COVID-19), caused by the late 2019 outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), causes a respiratory disease which could put myasthenia gravis (MG) patients at a greater risk of developing severe disease course, since infections and some drugs are a well-recognized trigger of symptom exacerbation in MG patients. Out of ten most commonly used past and present drugs used in COVID-19 treatment, two (quinolone derivatives and azithromycin) are known to worsen MG symptoms, whereas another two (tocilizumab and eculizumab) might have positive effect on MG symptoms. Colchicine, remdesivir, lopinavir, ritonavir and favipiravir seem to be safe to use, while data are insufficient for bamlanivimab, although it is also probably safe to use. Considering MG treatment options in patients infected with SARS-CoV-2, acetylcholine esterase inhibitors are generally safe to use with some preliminary studies even demonstrating therapeutic properties in regard to COVID-19. Corticosteroids are in general safe to use, even recommended in specific circumstances, whereas other immunosuppressive medications (mycophenolate mofetil, azathioprine, cyclosporine, methotrexate) are probably safe to use. The only exception is rituximab since the resulting B cell depletion can lead to more severe COVID-19 disease. Concerning plasmapheresis and intravenous immunoglobulins, both can be used in COVID-19 while taking into consideration thromboembolic properties of the former and hemodynamic disturbances of the latter. As current data suggest, all known COVID-19 vaccines are safe to use in MG patients.


Subject(s)
COVID-19 , Myasthenia Gravis , Antibodies, Monoclonal, Humanized , Antibodies, Neutralizing , COVID-19/complications , COVID-19/drug therapy , COVID-19 Vaccines , Humans , Myasthenia Gravis/complications , Myasthenia Gravis/therapy , SARS-CoV-2
4.
Clin Neurol Neurosurg ; 213: 107140, 2022 02.
Article in English | MEDLINE | ID: covidwho-1654200

ABSTRACT

OBJECTIVE: Recent studies suggest that the clinical course and outcomes of patients with coronavirus disease 2019 (COVID-19) and myasthenia gravis (MG) are highly variable. We performed a systematic review of the relevant literature with a key aim to assess the outcomes of invasive ventilation, mortality, and hospital length of stay (HLoS) for patients presenting with MG and COVID-19. METHODS: We searched the PubMed, Scopus, Web of Science, and MedRxiv databases for original articles that reported patients with MG and COVID-19. We included all clinical studies that reported MG in patients with confirmed COVID-19 cases via RT-PCR tests. We collected data on patient background characteristics, symptoms, time between MG and COVID-19 diagnosis, MG and COVID-19 treatments, HLoS, and mortality at last available follow-up. We reported summary statistics as counts and percentages or mean±SD. When necessary, inverse variance weighting was used to aggregate patient-level data and summary statistics. RESULTS: Nineteen studies with 152 patients (mean age 54.4 ± 12.7 years; 79/152 [52.0%] female) were included. Hypertension (62/141, 44.0%) and diabetes (30/141, 21.3%) were the most common comorbidities. The mean time between the diagnosis of MG and COVID-19 was7.0 ± 6.3 years. Diagnosis of COVID-19 was confirmed in all patients via RT-PCR tests. Fever (40/59, 67.8%) and ptosis (9/55, 16.4%) were the most frequent COVID-19 and MG symptoms, respectively. Azithromycin and ceftriaxone were the most common COVID-19 treatments, while prednisone and intravenous immunoglobulin were the most common MG treatments. Invasive ventilation treatment was required for 25/59 (42.4%) of patients. The mean HLoS was 18.2 ± 9.9 days. The mortality rate was 18/152 (11.8%). CONCLUSION: This report provides an overview of the characteristics, treatment, and outcomes of MG in COVID-19 patients. Although COVID-19 may exaggerate the neurological symptoms and worsens the outcome in MG patients, we did not find enough evidence to support this notion. Further studies with larger numbers of patients with MG and COVID-19 are needed to better assess the clinical outcomes in these patients.


Subject(s)
COVID-19/complications , COVID-19/therapy , Myasthenia Gravis/complications , Myasthenia Gravis/therapy , Adolescent , Adult , COVID-19/mortality , Child , Female , Hospitalization , Humans , Male , Middle Aged , Myasthenia Gravis/mortality , Respiration, Artificial , Survival Rate , Young Adult
5.
Neurol Sci ; 43(4): 2271-2276, 2022 Apr.
Article in English | MEDLINE | ID: covidwho-1636977

ABSTRACT

BACKGROUND: Myasthenia gravis (MG) is an autoimmune disorder of the neuromuscular junction that can be exacerbated by many viral infections, including COVID-19. The management of MG exacerbations is challenging in this scenario. We report 8 cases of MG exacerbation or myasthenic crisis associated with COVID-19 and discuss prognosis and treatment based on a literature review. RESULTS: Most patients were female (7/8), with an average age of 47.1 years. Treatment was immunoglobulin (IVIG) in 3 patients, plasma exchange (PLEX) in 2 patients, and adjustment of baseline drugs in 3. In-hospital mortality was 25% and 37.5% in 2-month follow-up. DISCUSSION: This is the largest case series of MG exacerbation or myasthenic crisis due to COVID-19 to this date. Mortality was considerably higher than in myasthenic crisis of other etiologies. Previous treatment for MG or acute exacerbation treatment did not seem to interfere with prognosis, although sample size was too small to draw definitive conclusions. Further studies are needed to understand the safety and effectiveness of interventions in this setting, particularly of PLEX, IVIG, rituximab, and tocilizumab.


Subject(s)
COVID-19 , Myasthenia Gravis , COVID-19/complications , Female , Humans , Middle Aged , Myasthenia Gravis/complications , Myasthenia Gravis/therapy , Plasma Exchange , SARS-CoV-2
6.
Muscle Nerve ; 65(4): 447-452, 2022 04.
Article in English | MEDLINE | ID: covidwho-1626917

ABSTRACT

INTRODUCTION/AIMS: Coronavirus disease-2019 (COVID-19) may have a more severe course in patients with myasthenia gravis (MG). We aimed to assess severity of the infection and factors contributing to its severity in a group of MG patients, most of whom were not hospitalized. METHODS: One hundred forty outpatients with MG followed between March 2020 and April 2021 were included in our study. Patients were asked to respond to a brief questionnaire in person, by telemedicine, or through electronic messages. RESULTS: Nineteen patients tested positive for COVID-19 by polymerase chain reaction. Two were asymptomatic. Of the 17 symptomatic patients, 11 had mild symptoms. They either had no treatment or received antivirals, antibiotics, and anticoagulants. Their myasthenia was well-controlled before infection and was unaffected by COVID-19. Three patients with moderate COVID-19 required hospitalization, but not intensive care, and had full recovery. Three other patients, the oldest in the cohort, had severe disease: One patient with a postsurgery myasthenic exacerbation before the infection needed intensive care without intubation, but recovered completely; two morbidly obese patients with comorbidities required intubation and died. Corticosteroids were increased in four of the six moderate/severely affected patients. Immunosuppressive (IS) agents were generally continued. Hydroxychloroquine (HCQ) for COVID-19 was used in one patient. DISCUSSION: Most patients had mild COVID-19 and all but two patients recovered. The design of the study made it possible to capture mild cases. Having well-controlled MG before infection and absence of comorbidities likely affected the course of the infection favorably. IS did not influence the progression.


Subject(s)
COVID-19 , Myasthenia Gravis , Obesity, Morbid , Ambulatory Care Facilities , Humans , Myasthenia Gravis/complications , Myasthenia Gravis/epidemiology , Myasthenia Gravis/therapy , SARS-CoV-2 , Treatment Outcome
7.
Neurol India ; 69(6): 1772-1776, 2021.
Article in English | MEDLINE | ID: covidwho-1606774

ABSTRACT

This report describes a patient with thymomatous myasthenia gravis (MG) with aplastic anemia in pharmacological remission and COVID-19 who developed respiratory failure in the course of the disease and reviews the published literature on this topic. Analysis of the clinical characteristics of the eight patients with MG including our patient suggests two possible mechanisms for respiratory failure: myasthenic crisis (MC) or pulmonary complications of COVID-19. Patients with MC were young women in high-grade MGFA Class whereas patients with respiratory failure due to pulmonary complications of COVID-19 were elderly men in pharmacological remission or MGFA Class I. These observations suggest that COVID-19, like other infections, may precipitate MC in patients with severe grade MG before COVID-19. The only differentiating feature between the two types of failure was severity myasthenic weakness. This clinical distinction has management implications. These observations need to be validated in a larger sample.


Subject(s)
COVID-19 , Myasthenia Gravis , Respiratory Insufficiency , Aged , Female , Humans , Male , Muscle Weakness , Myasthenia Gravis/complications , Respiratory Insufficiency/etiology , SARS-CoV-2
8.
Eur J Neurol ; 28(10): 3418-3425, 2021 10.
Article in English | MEDLINE | ID: covidwho-1605552

ABSTRACT

BACKGROUND AND PURPOSE: Myasthenia gravis (MG) patients could be a vulnerable group in the pandemic era of coronavirus 2019 (COVID-19) mainly due to respiratory muscle weakness, older age and long-term immunosuppressive treatment. We aimed to define factors predicting the severity of COVID-19 in MG patients and risk of MG exacerbation during COVID-19. METHODS: We evaluated clinical features and outcomes after COVID-19 in 93 MG patients. RESULTS: Thirty-five patients (38%) had severe pneumonia and we recorded 10 deaths (11%) due to COVID-19. Higher forced vital capacity (FVC) values tested before COVID-19 were shown to be protective against severe infection (95% CI 0.934-0.98) as well as good control of MG measured by the quantified myasthenia gravis score (95% CI 1.047-1.232). Long-term chronic corticosteroid treatment worsened the course of COVID-19 in MG patients (95% CI 1.784-111.43) and this impact was positively associated with dosage (p = 0.005). Treatment using azathioprine (95% CI 0.448-2.935), mycophenolate mofetil (95% CI 0.91-12.515) and ciclosporin (95% CI 0.029-2.212) did not influence the course of COVID-19. MG patients treated with rituximab had a high risk of death caused by COVID-19 (95% CI 3.216-383.971). Exacerbation of MG during infection was relatively rare (15%) and was not caused by remdesivir, convalescent plasma or favipiravir (95% CI 0.885-10.87). CONCLUSIONS: As the most important predictors of severe COVID-19 in MG patients we identified unsatisfied condition of MG with lower FVC, previous long-term corticosteroid treatment especially in higher doses, older age, the presence of cancer, and recent rituximab treatment.


Subject(s)
COVID-19 , Coronavirus Infections , Myasthenia Gravis , Aged , COVID-19/therapy , Humans , Immunization, Passive , Myasthenia Gravis/complications , Myasthenia Gravis/drug therapy , Myasthenia Gravis/epidemiology , SARS-CoV-2
12.
J Neuromuscul Dis ; 7(3): 361-364, 2020.
Article in English | MEDLINE | ID: covidwho-1372059

ABSTRACT

This is a brief report of a patient who has refractory Myasthenia Gravis, on multiple long-term immunosuppressive therapies and contracted COVID-19 during this 2020 pandemic. She was quarantined for total of 14 days and recovered successfully without any complications (no myasthenia exacerbation or crisis, no COVID-19 related complications), with no changes to her immunosuppressive therapy. Treatment of MG patients with COVID-19 needs to be tailored to individual patient.


Subject(s)
Betacoronavirus , Coronavirus Infections/complications , Immunosuppressive Agents/therapeutic use , Myasthenia Gravis/drug therapy , Pneumonia, Viral/complications , Adult , COVID-19 , Chronic Disease , Female , Humans , Myasthenia Gravis/complications , Pandemics , Precision Medicine/methods , Risk Factors , SARS-CoV-2 , Treatment Outcome
13.
BMJ Case Rep ; 14(8)2021 Aug 11.
Article in English | MEDLINE | ID: covidwho-1354559

ABSTRACT

We report on a unique case of a 7-year-old girl with new onset ocular myasthenia gravis shortly after recovery from multisystem inflammatory syndrome in children (MIS-C) temporally associated with SARS-CoV-2 infection. The diagnosis of myasthenia gravis was based on suggestive symptoms of fatigable bilateral orbital ptosis, diplopia, positive ocular cold compression test and serum acetylcholine receptor antibody positivity, as well as a favourable treatment response to pyridostigmine. The addition of corticosteroids and methotrexate resulted in complete resolution of the ocular signs.


Subject(s)
COVID-19 , Myasthenia Gravis , Child , Female , Humans , Myasthenia Gravis/complications , Myasthenia Gravis/diagnosis , Myasthenia Gravis/drug therapy , Receptors, Cholinergic , SARS-CoV-2 , Systemic Inflammatory Response Syndrome
14.
BMJ Case Rep ; 14(7)2021 Jul 21.
Article in English | MEDLINE | ID: covidwho-1320440

ABSTRACT

Several case reports of COVID-19 in patients with myasthenia gravis (MG) have been documented. However, new-onset autoimmune MG following COVID-19 has been reported very rarely. We report one such case here. A 65-year-old man presented to us with dysphagia 6 weeks following mild COVID-19. He was evaluated and diagnosed as antiacetylcholine receptor antibody (AchR) positive, non-thymomatous, generalised MG. He subsequently developed myasthenic crisis and improved after treatment with intravenous immunoglobulin, prednisolone and pyridostigmine. Systematic literature review showed eight more similar cases. Analysis of all cases including the one reported here showed these features: mean age 55.8 years, male gender (5), time interval between COVID-19 and MG (5-56 days), generalised (5), bulbar and/or ocular symptoms (4), anti-AchR antibodies (7) and antimuscle-specific kinase antibodies (2). All have improved with immunotherapy. Although, many hypothesis are proposed to explain causal relationship between the two, it could as well be sheer coincidence.


Subject(s)
COVID-19 , Myasthenia Gravis , Aged , Humans , Male , Middle Aged , Myasthenia Gravis/complications , Myasthenia Gravis/diagnosis , Myasthenia Gravis/drug therapy , Pyridostigmine Bromide/therapeutic use , Receptors, Cholinergic , SARS-CoV-2
15.
Eur J Neurol ; 28(10): 3418-3425, 2021 10.
Article in English | MEDLINE | ID: covidwho-1255377

ABSTRACT

BACKGROUND AND PURPOSE: Myasthenia gravis (MG) patients could be a vulnerable group in the pandemic era of coronavirus 2019 (COVID-19) mainly due to respiratory muscle weakness, older age and long-term immunosuppressive treatment. We aimed to define factors predicting the severity of COVID-19 in MG patients and risk of MG exacerbation during COVID-19. METHODS: We evaluated clinical features and outcomes after COVID-19 in 93 MG patients. RESULTS: Thirty-five patients (38%) had severe pneumonia and we recorded 10 deaths (11%) due to COVID-19. Higher forced vital capacity (FVC) values tested before COVID-19 were shown to be protective against severe infection (95% CI 0.934-0.98) as well as good control of MG measured by the quantified myasthenia gravis score (95% CI 1.047-1.232). Long-term chronic corticosteroid treatment worsened the course of COVID-19 in MG patients (95% CI 1.784-111.43) and this impact was positively associated with dosage (p = 0.005). Treatment using azathioprine (95% CI 0.448-2.935), mycophenolate mofetil (95% CI 0.91-12.515) and ciclosporin (95% CI 0.029-2.212) did not influence the course of COVID-19. MG patients treated with rituximab had a high risk of death caused by COVID-19 (95% CI 3.216-383.971). Exacerbation of MG during infection was relatively rare (15%) and was not caused by remdesivir, convalescent plasma or favipiravir (95% CI 0.885-10.87). CONCLUSIONS: As the most important predictors of severe COVID-19 in MG patients we identified unsatisfied condition of MG with lower FVC, previous long-term corticosteroid treatment especially in higher doses, older age, the presence of cancer, and recent rituximab treatment.


Subject(s)
COVID-19 , Coronavirus Infections , Myasthenia Gravis , Aged , COVID-19/therapy , Humans , Immunization, Passive , Myasthenia Gravis/complications , Myasthenia Gravis/drug therapy , Myasthenia Gravis/epidemiology , SARS-CoV-2
17.
Medicine (Baltimore) ; 100(18): e25701, 2021 May 07.
Article in English | MEDLINE | ID: covidwho-1216694

ABSTRACT

RATIONALE: Myasthenia gravis (MG) patients are at increased risk of COVID-19 infection and its complications due to chronic immunosuppression. COVID-19 infection can also increase the risk of myasthenia exacerbation. PATIENT CONCERNS: The patient presented with respiratory distress, fever and chills and was diagnosed with COVID-19 pneumonia. His past medical history includes seropositive generalized MG diagnosed in 2019, hypertension, atrial fibrillation and congestive heart failure with reduced ejection failure. DIAGNOSES: Refractory seropositive generalized MG having COVID-19 pneumonia and respiratory failure (needing mechanical ventilation) with sepsis. INTERVENTION: Use of intravenous remdesivir and dexamethasone and patient's myasthenic exacerbation (due to COVID-19 and its complications) was successfully treated with plasmapheresis. OUTCOMES: Patient was successfully weaned off ventilator to trach collar and was discharged to inpatient rehabiliation. He was followed up 1 month post hospital discharge and was on trach collar. LESSONS: This case report illustrates early use of the combination therapy might be beneficial in refractory myasthenia gravis cases even with chronic immunosuppression and severe COVID-19 infection.


Subject(s)
Adenosine Monophosphate/analogs & derivatives , Alanine/analogs & derivatives , Antiviral Agents/therapeutic use , COVID-19/drug therapy , Dexamethasone/therapeutic use , Glucocorticoids/therapeutic use , Myasthenia Gravis/complications , Pneumonia, Viral/drug therapy , Adenosine Monophosphate/therapeutic use , Aged , Alanine/therapeutic use , Drug Therapy, Combination , Humans , Male , Plasmapheresis , SARS-CoV-2
18.
Acta Neurol Scand ; 144(3): 334-340, 2021 Sep.
Article in English | MEDLINE | ID: covidwho-1207377

ABSTRACT

OBJECTIVE: To describe presenting symptoms, clinical outcomes, and therapeutic management of concurrent Coronavirus disease 2019 (COVID-19) infections in patients with a pre-existing myasthenia gravis (MG). METHODS: We conducted a retrospective study in patients with preexisting MG presenting with concurrent COVID-19 between September 21st and November 4th, 2020 when attending the emergency department or routine neurology consultation at the National Institute Mongi Ben Hamida of Neurology of Tunis, Tunisia. RESULTS: Five patients were identified. The Myasthenia Gravis Foundation of America scores (MGFA) prior to COVID-19 infection were class I in one patient, class II (IIa, IIb) in two patients, and class IIIb in one patient. Four patients had mild to moderate courses of COVID-19 infection. One patient presented a critical infection with acute respiratory disease syndrome (ARDS) requiring mechanical ventilation. Two of them also demonstrated signs of MG exacerbation requiring the use of intravenous immunoglobulin in one case. We maintained immunosuppressant therapy to MG in all our patients. All our patients received Azithromycin (AZM) as a part of specific drug treatment of COVID-19 infection. Outcome was favorable in 4 patients and rapidly fatal evolution was observed in the patient with ADRS. DISCUSSIONS AND CONCLUSION: The results from our study suggest that prior MG activity could partially influence the subsequent clinical outcomes. It emerged also that ongoing long-term immunosuppressive immunotherapy to MG should be maintained during the COVID-19 pandemic and that AZM can be used safely in MG patients and concurrent COVID-19 infection.


Subject(s)
COVID-19/complications , Myasthenia Gravis/complications , Adult , COVID-19/immunology , COVID-19/therapy , Female , Humans , Male , Middle Aged , Myasthenia Gravis/immunology , Myasthenia Gravis/therapy , Retrospective Studies , SARS-CoV-2 , Tunisia
19.
Pharmacotherapy ; 41(6): 546-550, 2021 06.
Article in English | MEDLINE | ID: covidwho-1173838

ABSTRACT

Myasthenia gravis and the associated pharmacologic management options could place patients at higher risk of contracting severe acute respiratory syndrome coronavirus 2 and exhibiting more severe manifestations of the novel coronavirus disease 2019 (COVID-19). Multiple agents have been studied for the management of the COVID-19, including remdesivir. To date, no published reports have evaluated the utilization of the antiviral remdesivir in patients with myasthenia gravis. We describe the first reported clinical course of three patients with myasthenia gravis who safely received remdesivir in combination with dexamethasone for the management of COVID-19.


Subject(s)
Adenosine Monophosphate/analogs & derivatives , Alanine/analogs & derivatives , Antiviral Agents/therapeutic use , COVID-19/complications , COVID-19/drug therapy , Myasthenia Gravis/complications , Myasthenia Gravis/drug therapy , Adenosine Monophosphate/therapeutic use , Adult , Aged , Alanine/therapeutic use , Anti-Inflammatory Agents/therapeutic use , Dexamethasone/therapeutic use , Fatal Outcome , Female , Humans , Male , Middle Aged , Treatment Outcome
20.
Acta Neurol Belg ; 121(4): 1039-1044, 2021 Aug.
Article in English | MEDLINE | ID: covidwho-1163201

ABSTRACT

Coronavirus disease 2019 (COVID-19), caused by the late 2019 outbreak of Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), causes a respiratory disease which could put myasthenia gravis patients at a greater risk of developing severe disease course. This paper presents a single-institution case series of hospitalized myasthenia gravis patients with COVID 19. We identified eight patients previously diagnosed with myasthenia gravis, four of whom presented with clear signs of myasthenia gravis symptom worsening on admission. No form of respiratory support was needed during the complete duration of stay for three patients, oxygen therapy was administered to two patients, while the remaining three patients required mechanical ventilation. Treatment was successful for seven patients, six of whom were discharged without any myasthenia gravis symptoms. One patient died after eleven days of intensive care unit treatment. Although treatment of patients with myasthenia gravis and COVID-19 patients is challenging, case series of myasthenia gravis patients with COVID-19 treated in our institution demonstrates relatively favorable treatment outcome. Our data seem to support the notion that immunosuppressive medication does not seem to result in worse outcomes. Our data also support the notion that intravenous immunoglobulin treatment is safe and should be administered to patients with myasthenia gravis and COVID-19 in case of myasthenia gravis worsening since benefits seem to greatly outweigh the risks.


Subject(s)
COVID-19/complications , Myasthenia Gravis/complications , Croatia , Female , Humans , Immunoglobulins, Intravenous/therapeutic use , Male , Middle Aged , Myasthenia Gravis/drug therapy , SARS-CoV-2
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