High Titers Research Articles

Newcastle disease vaccination program in broilers using a Apathogenic heat-resistant vaccine

The Newcastle disease (ND) is among the most common and deadly poultry diseases worldwide. Thermostable Newcastle disease virus (NDV) vaccines have been used widely to protect village chickens against Newcastle disease, due to their decreased dependence on cold chain for transport and storage. The Newcastle Disease V4 Heat-Resistant (NDV4HR) vaccine is an Apathogenic heat-resistant live vaccine that can induce immunity in chickens. A total of 60 one-day-old Arain local hybrid broilers divided into three groups of 20. Group A had the usual vaccination program in broiler flocks (7 Days old: B1 type by eye drop+ ND Killed vaccine by IM; 18 and 28 days old: Lasota strain orally), group B did not receive any vaccine, and Group C received orally the NDV4HR vaccine six times from the first day to the 35th day of rearing. All groups were nasally challenged with acute Newcastle virus (genotype VIId) on the 35th rearing day. Regarding HI serum antibody titer of the birds after the challenge, group B had the highest (7.12 ± 1.05), and Group C (6.37 ± 1.31) had a higher titer than Group A (6 ± 1.22). After the challenge with the Newcastle virus, the necropsy findings and clinical symptoms of the disease were almost similar in groups A and C. Group B showed the most signs with higher casualties than other groups. Regarding weight gain, Group C had the most weight at the end of the study (2204 ± 106). On the second day after the challenge, groups A and B had virus shedding through the trachea and cloaca; however, group B shed the virus only through the trachea. Furthermore, on the 7th day after the challenge, group A shed the virus through the cloaca, and group B through the trachea and the cloaca. At the same time, there was no virus shedding in group C. This study showed that V4HR vaccine provokes an antibody response that protects the birds against virulent virus challenge.

Pretransplant immunologic risk assessment in high baseline ABO antibody titers and donor-specific anti-human leukocyte antigen antibodies in ABO incompatible kidney transplantation

Pretransplant immunologic risk assessment in high baseline ABO antibody titers and donor-specific anti-human leukocyte antigen antibodies in ABO incompatible kidney transplantation

Investigation of Anti-SARS-CoV-2-specific IgG Levels in Breast Milk after Vaccination or COVID-19 Infection.

Newborns are vulnerable to all types of infections due to their developing immune system. To compensate for their immune immaturity, newborns rely on the passive transfer of antibodies through the placenta and own mother's breast milk (BM). In the present study, we investigated whether vaccination against SARS-CoV-2 leads to the presence of antibodies in BM. Furthermore, we compared the levels of SARS-CoV-2-specific anti-spike (anti-S) IgG antibodies in the BM of mothers who were vaccinated against Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) or had coronavirus disease 2019 (COVID-19) infection naturally or were vaccinated after natural infection. This was a prospective cohort study conducted in the Ondokuz Mayis University Faculty of Medicine. Forty-six mothers who had at least two doses of the BNT162b2 messenger RNA-based vaccine and/or had a history of symptomatic COVID-19 infection were included in the study. BM samples were analyzed by the Abbott Architect SARS-CoV-2 IgG II Quant kit following the manufacturer's instructions. Forty-six mothers with an average age of 29.7 ± 5.7 years participated the study: 18 (39.1%) had COVID-19 infection + BTN162b2 vaccine, 17 (37.0%) had BTN162b2 vaccine, and 11 (23.9%) had natural infection. The highest SARS-CoV-2-specific anti-S IgG antibody titers in BM were found in mothers who were vaccinated following the infection (anti-SARS-CoV-2 IgG: 32.48 ± 57.1 arbitrary units AU/mL). However, no significant difference in anti-SARS-CoV-2 antibody levels was observed between the three groups (p = 0.641). No antibody was detected in 15.2% of BM samples. Both vaccination and natural COVID-19 infection during pregnancy leads to the passive transfer of specific anti-SARS-CoV-2 IgG antibodies to BM. These results are important to overcome vaccine hesitancy and increase vaccination levels in expectant mothers. · We investigated the levels of SARS-CoV-2 antibodies in BM after natural infection and vaccination.. · Anti-SARS-CoV-2 IgG antibodies were detected in 39 (84.8%) BM samples.. · The highest titers in BM were found in mothers who were vaccinated following natural infection..

Preclinical evaluation of RQ3013, a broad-spectrum mRNA vaccine against SARS-CoV-2 variants

The global emergence of SARS-CoV-2 variants has led to increasing breakthrough infections in vaccinated populations, calling for an urgent need to develop more effective and broad-spectrum vaccines to combat COVID-19. Here we report the preclinical development of RQ3013, an mRNA vaccine candidate intended to bring broad protection against SARS-CoV-2 variants of concern (VOCs). RQ3013, which contains pseudouridine-modified mRNAs formulated in lipid nanoparticles, encodes the spike (S) protein harboring a combination of mutations responsible for immune evasion of VOCs. Here we characterized the expressed S immunogen and evaluated the immunogenicity, efficacy, and safety of RQ3013 in various animal models. RQ3013 elicited robust immune responses in mice, hamsters, and nonhuman primates (NHP). It can induce high titers of antibodies with broad cross-neutralizing ability against the wild-type, B.1.1.7, B.1.351, B.1.617.2, and the newly emerging Omicron variants. In mice and NHP, two doses of RQ3013 protected the upper and lower respiratory tract against infection by SARS-CoV-2 and its variants. Furthermore, our safety assessment of RQ3013 in NHP showed no observable adverse effects. These results provide strong support for the evaluation of RQ3013 in clinical trials and suggest that it may be a promising candidate for broad protection against COVID-19 and its variants.

Albumin from sera of rheumatoid arthritis patients share multiple biochemical, biophysical and immunological properties with in vitro generated glyco-nitro-oxidized-albumin

The purpose of the present study is to explore the effects of endogenous stressors on structure and function of rheumatoid arthritis (RA) patients’ albumin. In contrast to glycated-albumin or nitro-oxidized-albumin, high titre antibodies against glyco-nitro-oxidized-albumin were found in the sera of RA patients. Also, compared to the other two modified forms of albumin, glyco-nitro-oxidized-albumin showed highest percent inhibition. Albumin isolated from RA patients’ sera displayed hyperchromicity and quenching of tyrosine and tryptophan fluorescence. Fluorescence spectroscopy studies also revealed the presence of dityrosine and advanced glycation end products in RA patient’s albumin. RA patients’ albumin showed weaker binding with 1-anilinonaphthalene-8-sulfonic acid dye. Secondary structure alterations were demonstrated by circular dichroism and Fourier transform infrared spectroscopy. Biochemical investigations revealed substantial decline in the availability of free side chains of amino acid residues; increased carbonyls and decreased sulfhydryls in RA patients’ albumin. The functional impairment in RA patients’ albumin was revealed by their low binding with bilirubin and cobalt. Liquid chromatography mass spectrometry analysis revealed the presence of Nε-(carboxymethyl) lysine and 3-nitrotyrosine in RA patients’ albumin. The amyloidogenic aggregation of RA patients’ albumin was confirmed by Congo red absorption and thioflavin-T fluorescence assays. The morphology of the aggregates was visualized under scanning and transmission electron microscope. From the above findings, we inferred that endogenous stress in RA patients have modified albumin and produce structural/functional abnormalities. Also, the presence of anti-glyco-nitro-oxidized-albumin antibodies along with other clinical features may be used as biomarker for the diagnosis and assessment of treatment responses in RA patients. Communicated by Ramaswamy H. Sarma

Atypical Ebola Virus Disease in a Rhesus Macaque.

Ebola virus (EBOV)-Makona infected more than 30 000 people from 2013 to 2016 in West Africa, among them many health care workers including foreign nationals. Most of the infected foreign nationals were evacuated and treated in their respective home countries, resulting in detailed reports of the acute disease following EBOV infection as well as descriptions of symptoms now known as post-Ebola syndrome, which occurred months after the infection. Symptoms associated with this syndrome include uveitis and neurological manifestations. In 1 of our EBOV-Makona nonhuman primate (NHP) studies, 1 NHP was euthanized on day 28 after infection having completely recovered from the acute disease. During convalescence, this NHP developed neurological signs and acute respiratory distress requiring euthanasia. The organ tropism had changed with high virus titers in lungs, brain, eye, and reproductive organs but no virus in the typical target organs for acute EBOV infection. This in part reflects sequelae described for EBOV survivors albeit developing quicker after recovery from acute disease.

Cold Agglutinin Disease: Challenges in the Serological Workup and Management

Abstract Cold agglutinin disease (CAD) is a rare autoimmune hemolytic anemia associated with autoantibodies that react mainly in the cold. CAD can interfere with immunohematology tests. We report two cases of CAD presented initially as blood grouping discrepancy leading us to further workup. Case I was a 61-year-old female with symptoms of weakness, bone pain, and difficulty breathing with laboratory findings of high cold agglutinin titers of 1:1024. Bone marrow aspirate revealed a mixture of >30% plasma cell and plasma blast. Bone marrow biopsy revealed an interstitial, focal, and nodular pattern of involvement by plasma cells, leading to the diagnosis of plasma cell dyscrasias. Case II analyzed was benign CAD with cold agglutinin titers of 1:64, with no other abnormality. Both patients were managed successfully. We report serological and clinical findings of two CAD cases with similar serological findings but different clinical outcomes.

PIVKA‑II is associated with liver function, bone metabolism, and muscle function in patients with liver disease.

Protein induced by vitamin K (VK) absence-II (PIVKA-II) is a sensitive marker for diagnosing hepatoma but is occasionally detected in patients without hepatoma Here, the clinical significance of serum PIVKA-II levels in patients who were not administered warfarin and did not have hepatoma or liver disease were evaluated. As VK is related to muscle and bone metabolism, PIVKA-II and clinical factors related to bone and muscle were compared. A total of 441 patients with various liver diseases were evaluated. Of these, 236 patients were female. Clinical factors and anthropometric measurements were obtained for each participant during outpatient visits. Among the clinical factors, type I procollagen N-propeptide (P1NP), a low titer of undercarboxylated osteocalcin (ucOC), and 25(OH) vitamin D (VD) were used as bone metabolic markers, and SARC-F and grip strength were used as muscle-related markers. Serum PIVKA-II levels above the upper limit were associated with Child B/C (Child-Pugh score), high titers of total P1NP, and low titers of ucOC in females, and alcohol-related liver disease and low VD in males. The titer of PIVKA-II were associated with immunoglobulin (Ig) A and prothrombin time (PT)-international normalized ratio (INR) in females, and fibrosis-4-4, IgG, total bilirubin, PT-INR, and SARC-F in males. Elevated PIVKA-II levels were associated with abnormal bone physiology in females, weak muscles in males, and severe liver disease in both sexes. Assessing PIVKA-II may assist in evaluating the clinical and bone-muscle metabolic stages in liver disease. Nutrition and supplementation with fat-soluble vitamins, including VK and VD may thus serve as a potential method to alleviate or prevent bone-muscle pathophysiology in patients with liver disease.

Rare Association of Takayasu Arteritis and Antiphospholipid Syndrome with Severe Thrombocytopenia

The association of Takayasu arteritis (TA) with antiphospholipid syndrome (APS) has rarely been described in the literature. This paper reports the first documented case of TA and APS in a 24-year-old woman in Bangladesh. This patient had claudication pain in upper and lower limbs for four years and Raynaud’s phenomenon in right hand for eight months. During the course of her illness, she suddenly developed deep vein thrombosis in left superficial femoral, popliteal and posterior tibial veins. Conventional angiography revealed total occlusion of right subclavian artery and 60-70% stenosis of right common iliac artery. CT angiography of right upper limb also supported these findings along with increased wall thickening in the 2nd part of right subclavian artery and moderate narrowing of the 1st part of right axillary artery. She had persistently positive anticardiolipin antibodies in high titers, positive lupus anticoagulant (LA), prolonged activated partial thromboplastin time (APTT) and severe thrombocytopenia. We started high dose prednisolone (1mg/kg daily). Her platelet count increased rapidly. Based on review of 10 case reports, we considered the rare association of TA and APS. Both conditions should be determined promptly for the sake of early institution of the appropriate therapy. Bangabandhu Sheikh Mujib Med. Coll. J. 2023;2(1): 59-63

Does the Presence or a High Titer of Yellow Fever Virus Antibodies Interfere with Pregnancy Outcomes in Women with Zika Virus Infection?

Zika virus (ZIKV) and yellow fever virus (YFV) originated in Africa and expanded to the Americas, where both are co-circulated. It is hypothesized that in areas of high circulation and vaccination coverage against YFV, children of pregnant women have a lower risk of microcephaly. We evaluated the presence and titers of antibodies and outcomes in women who had ZIKV infection during pregnancy. Pregnancy outcomes were classified as severe, moderate, and without any important outcome. An outcome was defined as severe if miscarriage, stillbirth, or microcephaly occurred, and moderate if low birth weight and/or preterm delivery occurred. If none of these events were identified, the pregnancy was defined as having no adverse effects. A sample of 172 pregnant women with an acute ZIKV infection confirmed during pregnancy were collected throughout 2016. About 89% (150 of 169) of them presented immunity against YFV, including 100% (09 of 09) of those who had severe outcomes, 84% (16 of 19) of those who had moderate outcomes, and 89% (125 of 141) of those who had non-outcomes. There was no difference between groups regarding the presence of anti-YFV antibodies (p = 0.65) and YFV titers (p = 0.6). We were unable to demonstrate a protective association between the presence or titers of YFV antibodies and protection against serious adverse outcomes from exposure to ZIKV in utero.

Assessment of the State of Antidiphtheria Immunity in Different Age Groups of the Population of the Russian Federation Based on Seromonitoring Data for 2015–2021

Relevance. Diphtheria is one of the dangerous diseases that threaten human life. Immunization is the only means of creating a favorable epidemiological situation. Ensuring the immunity of the population to this infection prevents the occurrence of the rise and spread of diphtheria among the population. One of the ways to assess the state of specific immunity to diphtheria is annual serological monitoring. Aims. Assessment of the state of antidiphtheria immunity in different age groups of the population of the Russian Federation based on seromonitoring data for 2015–2021. Materials and methods. The data of serological monitoring of diphtheria and the state of vaccination of the population by indicator groups from 85 subjects of the Russian Federation in 2015–2021 were analyzed. In just 7 years, in terms of seromonitoring in Russia, 439,874 people were examined for the presence of antibodies to diphtheria, including 73,559 children in the age group of 3–4 years, 74,424 adolescents aged 15–17 years, and 291,891 adults aged 18 and older. During the same period, 12,154 controlled blood sera from 20 subjects of the Russian Federation were examined. In the age group of 3–4 years – 1834 children, 15–17 years – 1366 teenagers, 18–29 years – 1711 adults, 30–39 years – 1724 adults, 40–49 years – 1918 adults, 50–59 years – 1798 adults, 60 years and older – 1803 adults. The presence of antibodies to diphtheria was determined in the course of passive hemagglutination. The protective titers of antibodies were taken to be dilution of sera with a titer of 1:20 or more, the intensity of anti-diphtheria immunity was assessed as follows: antibody titers in dilution of sera 1:20–1:40 – low titers, 1:80–1:160 – medium and 1:320 or more – high titers of antibodies of JSC «Biomed» im. I. I. Mechnikov. Results and discussion. In Russia, improving the surveillance system for diphtheria continues to be an urgent task of the private epidemiology of this infection. The data obtained during the study allowed us to characterize the population immunity of the population of the Russian Federation, as a whole across the country, as sufficient to prevent the occurrence and spread of diphtheria. The main factor contributing to this was the long-term high coverage of vaccinations against diphtheria of persons in the decreed age groups (more than 95% of those vaccinated from the number of persons to be vaccinated in each age group). When comparing the level of immunization of the population depending on age, it was found that the percentage of vaccinated persons in the age group of people 50 years and older was statistically significantly lower than in the group of people younger than 50 years. The data of serological studies in Russia as a whole showed a high proportion of people with protective titers of antibodies to diphtheria in each age group – 3–4 years – 96.7 ± 0.1%, 15–17 years – 98.3 ± 0.1%, 18–29 years – 96,0 ± 0,4% – 97,5 ± 0,3%, 30–39 – 95,9 ± 0,4% – 97,5 ± 0.3%, 40–49 years old – 95,7 ± 0,4% – 97,7 ± 0,3%. The largest proportion of seronegative individuals is detected among groups aged 50–59 years and 60 years and older. The analysis of the share of seronegative persons in certain subjects of the Russian Federation revealed that the number of territories with a share of seronegative persons of more than 10% in the age groups of 50–59 years and 60 years and older was significantly higher than in other age groups. Conclusion. Thus, when planning companies for «сatch-up» immunization, it is advisable to provide mandatory revaccination of seronegative persons over 50 years of age. Timely revaccination according to the National Vaccination Calendar, tracking the proportion of persons susceptible to diphtheria in all indicator groups, followed by revaccination of seronegative persons in older ages will allow maintaining a high level of population immunity (more than 95%) and ensuring a consistently low incidence of diphtheria in the country for many years.

Prospective Evaluation of High Titer Autoantibodies and Fetal Home Monitoring in the Detection of Atrioventricular Block Among Anti-SSA/Ro Pregnancies.

This prospective study of pregnant patients, Surveillance To Prevent AV Block Likely to Occur Quickly (STOP BLOQ), addresses the impact of anti-SSA/Ro titers and utility of ambulatory monitoring in the detection of fetal second-degree atrioventricular block (AVB). Women with anti-SSA/Ro autoantibodies by commercial testing were stratified into high and low anti-52-kD and/or 60-kD SSA/Ro titers applying at-risk thresholds defined by previous evaluation of AVB pregnancies. The high-titer group performed fetal heart rate and rhythm monitoring (FHRM) thrice daily and weekly/biweekly echocardiography from 17-26 weeks. Abnormal FHRM prompted urgent echocardiography to identify AVB. Anti-52-kD and/or 60-kD SSA/Ro met thresholds for monitoring in 261 of 413 participants (63%); for those, AVB frequency was 3.8%. No cases occurred with low titers. The incidence of AVB increased with higher levels, reaching 7.7% for those in the top quartile for anti-60-kD SSA/Ro, which increased to 27.3% in those with a previous child who had AVB. Based on levels from 15 participants with paired samples from both an AVB and a non-AVB pregnancy, healthy pregnancies were not explained by decreased titers. FHRM was considered abnormal in 45 of 30,920 recordings, 10 confirmed AVB by urgent echocardiogram, 7 being second-degree AVB, all <12 hours from normal FHRM and within another 0.75 to 4 hours to echocardiogram. The one participant with second/third-degree and two participants with third-degree AVB were diagnosed by urgent echocardiogram >17 to 72 hours from an FHRM. Surveillance echocardiograms detected no AVB when the preceding interval FHRM recordings were normal. High-titer antibodies are associated with an increased incidence of AVB. Anti-SSA/Ro titers remain stable over time and do not explain the discordant recurrence rates, suggesting that other factors are required. Fetal heart rate and rhythm (FHRM) with results confirmed by a pediatric cardiologist reliably detects conduction abnormalities, which may reduce the need for serial echocardiograms.

Depressive symptoms and anti-N-methyl-D-aspartate-receptor GluN1 antibody seropositivity in the PROSpective cohort with incident stroke

BackgroundAnti-NMDA-receptor GluN1 antibodies (NMDAR1-abs) are present in an autoimmune encephalitis with severe neuropsychiatric symptoms. We aimed to estimate the impact of serum NMDAR1-abs on depressive symptoms years after first-ever ischemic stroke (IS). MethodsData were used from the PROSpective Cohort with Incident Stroke-Berlin (PROSCIS-B; NCT01363856). Serum NMDAR1-abs (IgM/IgA/IgG) were measured within 7 days after IS using cell-based assays. We defined seropositivity as titers ≥1:10, thereof low titers as ≤1:100 and high titers as >1:100. We used the Center for Epidemiological Studies–Depression (CES-D) scale to measure depressive symptoms at year one, two and three following IS. We calculated crude and confounder adjusted weighted generalized linear models to quantify the impact of NMDAR1-abs on CES-D assessed at three annual time-points. ResultsNMDAR1-abs were measured in 583 PROSCIS-B IS patients (mean age = 67 [SD = 13]; 42%female; median NIHSS = 2 [IQR = 1–4]) of whom 76 (13%; IgM: n = 49/IgA: n = 43/IgG: n = 2) were seropositive, 55 (9%) with low and 21 (4%) with high titers. CES-D regarded over all follow-up time-points was higher in seropositive patients (βcrude = 2.56 [95%CI = −0.34 to 5.45]; βadjusted = 2.26 [95%CI = −0.68 to 5.20]) and effects were highest in patients with high titer (low titers: βcrude = 1.42 [95%CI = −1.79 to 4.62], βadjusted = 0.53 [95%CI = −2.47 to 3.54]; high titers: βcrude = 5.85 [95%CI = 0.20 to 11.50]; βadjusted = 7.20 [95%CI = 0.98 to 13.43]). ConclusionPatients with serum NMDAR1-abs (predominantly IgM&IgA) suffer more severe depressive symptoms after mild-to-moderate IS compared to NMDAR1-abs seronegative patients.

Association between anti-cardiac autoantibodies in acute myocarditis and downstream clinical outcomes

Abstract Introduction Myocarditis is an inflammatory heart muscle disease that can result in cardiac dysfunction and arrhythmias. Its aetiology includes infectious, toxic or autoimmune causes. Myocarditis is responsible for sudden cardiac death in young adults (1) and can result in heart failure due to dilated cardiomyopathy in 20% of patients (2). The pathogenesis of myocarditis may involve immune system dysregulation leading to a cascade of inflammation and myocardial damage (3). For those patients that rapidly progress to end-stage dilated cardiomyopathy, an important dilemma is whether there is any prospect of heart function recovery. Therefore, there is an unmet need for a better understanding of key causal pathways and for developing a biological signature of disease progression. Hypothesis The progression of acute myocarditis depends on a persistent adaptive immune response against the heart, leading to ongoing inflammation, myocardial fibrosis and ultimately tissue destruction and remodelling. Aims We sought to explore anti-cardiac IgG serum levels, their temporal profile throughout follow up and their association with clinical and imaging data, in patients presenting with acute myocarditis. Methods We prospectively recruited 80 patients with a clinical diagnosis of acute myocarditis. We measured their total anti-cardiac IgG levels at admisssion, 3- and 12-months follow-up using an enzyme-linked immunosorbent assay (ELISA). We explored the association between the anti-cardiac IgG levels, cardiac MRI parameters (CMR) and clinical data. Results We detect an adaptive immune response against the heart in a subgroup of patients with acute myocarditis. 20% of patients develop high systemic titres of anti-cardiac IgG (OD threshold &amp;gt;0.65), within the first 7 days of presentation (Figure 1a). These patients continue to have persistent and significantly higher anti-cardiac IgG levels at 12 months (Figure 1b). Importantly, patients that mount a strong initial adaptive immune response against the heart, are more likely to show evidence of persistent oedema on CMR at 12 months(Figure 1c). Clinical event rates in our cohort have remained low at five-year follow-up. A small signal exists to indicate potentially higher rates of persistent symptoms (70% vs 47%), hospitalisations for heart failure (12% vs 8%) and new heart failure diagnosis (18% vs 11%), in patients that mount a strong adaptive immune response compared to those that do not. Conclusion A subset of patients with acute myocarditis develops a strong and persistent adaptive immune response against the heart. This may be associated with more myocardial oedema on cardiac MRI, persistent symptoms and higher rates of heart failure diagnoses and hospitalisations. Further work Plasma proteomics (targeted and un-targeted discovery) with validation in myocardial tissue. Analysis of CMR data with a focus on fibrosis assessment. ELISA for IgG subtypes and total IgG against myosin and troponin I.

Chronic active Epstein-Barr virus infection mimicking tuberculosis - A diagnostic enigma.

Chronic active EBV infection is a rare disorder prone for misdiagnosis. They present with a wide range of symptoms from indolent to aggressive clinical course. Clinico-pathological correlation with confirmation by ancillary techniques is inevitable to diagnose this disease. We present a case of a 29-year-old male with fever, weight loss, and lymphadenopathy for 6 months. Lymph node biopsy showed occasional granuloma with preserved architecture. Suspected to have tuberculosis, he received antitubercular treatment (ATT) with no response for 3 months. Subsequently, additional workup showed many EBV-positive cells in sinusoids with high serum EBV titer, confirming the difficult diagnosis of CAEBV. The present case highlights the difficulty in the diagnosis of this entity and also emphasizes the necessity to recognize this disorder in countries endemic for tuberculosis, as it is no longer bound by ethnicity and geographical boundaries.

Probable Anti-Glutamate Decarboxylase 65 (Gad65) Antibody-Associated Cerebellar Ataxia. Clinical Case Report and Literature Review

Introduction. As more information about cerebellar ataxia induced by anti- GAD65 antibodies is accumulated, cerebellar dysfunction is increasingly associated with autoimmune causes. The prevalence of other neurological syndromes associated with anti- GAD65 antibodies has been estimated, but the occurrence of cerebellar ataxia of the same cause has not yet been reported. The clinical presentation and management of anti-GAD65 antibodies induced cerebellar ataxia are currently known only from single cases and small series reports.&#x0D; Case report. We present a clinical case of a 52-year-old woman who was admitted to the hospital due to dizziness, impaired coordination, occasional choking, and slurred speech. Diagnostic procedures were performed, in which anti-GAD65 antibodies and atrophic changes in the upper parts of the cerebellum were detected on brain MRI, which led to a possible diagnosis of anti-GAD65 antibody-associated cerebellar ataxia.&#x0D; Discussion and literature review. Patients tend to be women in their 60s with clinical symptoms such as gait and posture ataxia. Most patients present with nystagmus, dysarthria, and limb ataxia. To make the diagnosis, it is crucial to detect high titers of anti-GAD65 antibodies, do intrathecal anti-GAD65 antibody synthesis, and perform a brain MRI, which may reveal atrophy of the cerebellum as the disease progresses. Corticosteroids are one of the recommended treatment methods, which were effective in our case. Maintenance therapy is essential to prevent relapse of the disease.

High titer (>100 g/L) ethanol production from pretreated corn stover hydrolysate by modified yeast strains

Developing a reliable lignocellulose pretreatment method to extract mixed sugars and engineering efficient strains capable of utilizing xylose are crucial for advancing cellulosic ethanol production. In this study, chemical and characterization analyses revealed that alkali cooking can significantly remove lignin from lignocellulose crops. The highest amount of mixed sugar was obtained from corn stover hydrolysates with a 15 % solid loading. Our genetically engineered yeast strain ΔsnR4, derived from a well-staged WXY70, demonstrated excellent performance in low 10 % solids loading corn stover hydrolysate, producing a high ethanol yield of 0.485 g/g total sugars. When a combined NaOH-ball milling pretreatment strategy was applied at high solids loading, ΔsnR4 exhibited the maximum ethanol titer of 110.9 g/L within 36 h, achieving an ethanol yield of 92.9 % theoretical maximum. Therefore, ΔsnR4 is highly compatible with high solid loading NaOH-ball milling pretreatment, making it a potential candidate for industrial cellulosic ethanol.

Abstract 13671: A Case of Diffuse Alveolar Hemorrhage While on Systemic Anticoagulation for Impella

A 57 year old male with PMH of non-ischemic cardiomyopathy (EF 30%), atrial fibrillation, and VT s/p ICD was admitted to the cardiac intensive care unit for cardiogenic shock. His condition deteriorated necessitating Impella 5.5 implantation as bridge to cardiac transplant. Systemic heparin was started per Impella protocol with a lower anti-Xa goal of 0.2 - 0.3 given post-op bleeding at the R axillary Impella site. Five days post Impella placement he developed bloody secretions from the endotracheal tube requiring multiple blood transfusions and discontinuation of systemic heparin, while maintaining heparin purge. Labs were notable for anti-Xa 0.22, hemoglobin 9.7, and platelets 221. CT Chest demonstrated parenchymal opacities concerning for diffuse alveolar hemorrhage (DAH) which was confirmed via bronchoalveolar lavage; he had no known history of lung disease. High-dose corticosteroids were administered as a high ANA titer (1:128) and low C3 suggested capillaritis. However, decreasing ANA (1: 320) and a negative autoimmune workup decreased the likelihood of a rheumatologic cause for DAH and steroids were discontinued. DAH secondary to acquired von Willebrand syndrome (aVWS) was considered, but factor VIII and vWF activity levels were normal. The bloody secretions improved with inhaled thromboxane and systemic heparin was restarted. Orthotopic heart and kidney transplant was performed with subsequent Impella removal 104 days after Impella placement. Discussion: We present the first documented case of DAH with isolated Impella 5.5 support. Previously reported cases of DAH post-Impella were observed in patients with concurrent ECMO or RVAD. As autoimmune and coagulopathic causes of DAH were excluded, bland DAH secondary to systemic anticoagulation is most likely in this case. This raises a question about how to balance the risk of bleeding with the risk of device thrombosis, stroke and embolic events and when to switch from heparin to sodium bicarbonate purge.

Abstract 15936: Prospective Evaluation of Anti-Ro Pregnancies Supports the Utility of Ambulatory Fetal Heart Rate Monitoring in Detecting Fetal Atrioventricular Block

Introduction:: The management of anti-Ro/SSA positive pregnancies is challenging because fetal atrioventricular (AV) block is rare and fetal echo seldom detects reversible injury. Hypothesis: We hypothesized that ambulatory fetal heart rate/rhythm monitoring improves detection of fetal AV block when compared with echo. Methods: Anti-Ro antibody subjects from 21 US centers at less than 20 weeks of gestation were risk stratified into high and low anti-Ro60 and 52 titers at the New York University (NYU) core laboratory. Low titer was defined by previous NYU data demonstrating no fetal AV block in subjects with anti-Ro60 and 52 less than 1000 arbitrary units per mL. High titer subjects performed 3x/day fetal heart rate/rhythm monitoring and underwent weekly or bi-weekly surveillance fetal echos at 18-26 weeks of gestation. Low titer subjects underwent site-specific surveillance. Abnormal fetal heart rate/rhythm monitoring prompted urgent diagnostic fetal echo. All infants received electrocardiograms. Results: We enrolled 405 subjects (fig). None of the 150 (37%) low titer subjects developed fetal AV block. Of 255 high titer subjects, 241 completed surveillance. High titer subjects monitored 44,187 times; 37 (0.08% ) monitoring episodes were deemed abnormal. Urgent fetal echos of the 37 diagnosed 9 with fetal AV block (7 with 2° AV block), 14 with atrial ectopy and 14 with normal rhythms. No fetal AV block, endocardial fibroelastosis or cardiac dysfunction was detected in the 871 surveillance fetal echoes of high titer subjects following normal FHRM. No fetus developed AV block after the surveillance period; one baby had transient prolonged PR interval, which resolved spontaneously. Conclusions: This prospective study suggests that ambulatory fetal heart rate/rhythm FHRM is superior to echo surveillance for fetal AV block, and antibody titers are useful in risk stratification of anti-Ro positive pregnancies.

Production and characterization of egg yolk antibodies against the ZIKV NS2B expressed in Nicotiana benthamiana

The emergence of Zika virus (ZIKV) and its associated neonatal and congenital complications pose a threat to global health, particularly in tropical and subtropical regions with co-circulation of related flaviviruses and intense vector proliferation. Diagnosis of ZIKV by RT-PCR is limited to the viraemic phase and is not always accessible in low-income tropical settings, while serological tests often show cross-reactivity with other flaviviruses. Given the similarity of ZIKV symptoms to those of other arboviruses, but the different prognosis and risks, it is important to develop specific and accessible diagnostic tools. Egg yolk antibodies (IgY) were obtained from Leghorn laying hens immunized with recombinant ZIKV NS2B protein produced in agroinfiltrated Nicotiana benthamiana. After three immunizations, total IgY was recovered from the eggs by the 20% ammonium sulfate precipitation method. After characterisation by SDS-PAGE, dot blotting and ELISA, the IgY was adsorbed to dengue virus (DENV) from cell culture supernatants and tested for its ability to specifically detect ZIKV-positive sera samples. High yield and purity were observed on SDS-PAGE for polyclonal IgY, which reacted with NS2B at high titres in ELISA and detected both NS2B and ZIKV in dot blotting. However, a cross-reaction with DENV was observed and the anti-NS2B IgY was unable to discriminate ZIKV from DENV positive sera samples, even after adsorption with DENV. This is probably due to the phylogenetic relationship of the viruses and the shared identity of their proteins.