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Neurology(R) Neuroimmunology &... Jul 2024Patients with ongoing seizures are usually not allowed to drive. The prognosis for seizure freedom is favorable in patients with autoimmune encephalitis (AIE) with...
BACKGROUND AND OBJECTIVES
Patients with ongoing seizures are usually not allowed to drive. The prognosis for seizure freedom is favorable in patients with autoimmune encephalitis (AIE) with antibodies against NMDA receptor (NMDAR), leucine-rich glioma-inactivated 1 (LGI1), contactin-associated protein-like 2 (CASPR2), and the gamma-aminobutyric-acid B receptor (GABAR). We hypothesized that after a seizure-free period of 3 months, patients with AIE have a seizure recurrence risk of <20% during the subsequent 12 months. This would render them eligible for noncommercial driving according to driving regulations in several countries.
METHODS
This retrospective multicenter cohort study analyzed follow-up data from patients aged 15 years or older with seizures resulting from NMDAR-, LGI1-, CASPR2-, or GABAR-AIE, who had been seizure-free for ≥3 months. We used Kaplan-Meier (KM) estimates for the seizure recurrence risk at 12 months for each antibody group and tested for the effects of potential covariates with regression models.
RESULTS
We included 383 patients with NMDAR-, 440 with LGI1-, 114 with CASPR2-, and 44 with GABAR-AIE from 14 international centers. After being seizure-free for 3 months after an initial seizure period, we calculated the probability of remaining seizure-free for another 12 months (KM estimate) as 0.89 (95% confidence interval [CI] 0.85-0.92) for NMDAR, 0.84 (CI 0.80-0.88) for LGI1, 0.82 (CI 0.75-0.90) for CASPR2, and 0.76 (CI 0.62-0.93) for GABAR.
DISCUSSION
Taking a <20% recurrence risk within 12 months as sufficient, patients with NMDAR-AIE and LGI1-AIE could be considered eligible for noncommercial driving after having been seizure-free for 3 months.
Topics: Humans; Female; Male; Adult; Intracellular Signaling Peptides and Proteins; Autoantibodies; Middle Aged; Encephalitis; Retrospective Studies; Receptors, GABA-B; Recurrence; Nerve Tissue Proteins; Young Adult; Membrane Proteins; Receptors, N-Methyl-D-Aspartate; Seizures; Hashimoto Disease; Aged; Adolescent; Follow-Up Studies; Proteins; Cohort Studies
PubMed: 38838283
DOI: 10.1212/NXI.0000000000200225 -
BMC Research Notes Jun 2024Fourth-generation HIV Ag/Ab Combo assay is used for HIV screening of blood for transfusion in developing countries, however, the sensitivity of the assay is questionable...
OBJECTIVE
Fourth-generation HIV Ag/Ab Combo assay is used for HIV screening of blood for transfusion in developing countries, however, the sensitivity of the assay is questionable during the acute phase of HIV infection. Thus, the study aimed to determine the effect of combining centrifugation with HIV-1 virion lysis on the sensitivity of the fourth-generation HIV Ag/Ab combo assay.
RESULTS
When the 50 HIV-1 antibody-negative samples were run on the fourth-generation HIV Ag/Ab combo assay, 8 (16%) were positive following centrifugation, 13 (26%) were positive following lysis while 25 (50%) were positive after combining centrifugation with HIV-1 virion lysis.
Topics: HIV-1; Humans; Centrifugation; HIV Infections; HIV Antibodies; Sensitivity and Specificity; Virion; HIV Antigens
PubMed: 38835056
DOI: 10.1186/s13104-024-06810-y -
International Journal of Nanomedicine 2024Prostate cancer (PC) is the second most common cancer and the fifth most frequent cause of cancer death among men. Prostate-specific membrane antigen (PSMA) expression...
INTRODUCTION
Prostate cancer (PC) is the second most common cancer and the fifth most frequent cause of cancer death among men. Prostate-specific membrane antigen (PSMA) expression is associated with aggressive PC, with expression in over 90% of patients with metastatic disease. Those characteristics have led to its use for PC diagnosis and therapies with radiopharmaceuticals, antibody-drug conjugates, and nanoparticles. Despite these advancements, none of the current therapeutics are curative and show some degree of toxicity. Here we present the synthesis and preclinical evaluation of a multimodal, PSMA-targeted dendrimer-drug conjugate (PT-DDC), synthesized using poly(amidoamine) (PAMAM) dendrimers. PT-DDC was designed to enable imaging of drug delivery, providing valuable insights to understand and enhance therapeutic response.
METHODS
The PT-DDC was synthesized through consecutive conjugation of generation-4 PAMAM dendrimers with maytansinoid-1 (DM1) a highly potent antimitotic agent, Cy5 infrared dye for optical imaging, 2,2',2"-(1,4,7-triazacyclononane-1,4,7-triyl)triacetic acid (NOTA) chelator for radiolabeling with copper-64 and positron emission tomography tomography/computed tomography (PET/CT), lysine-urea-glutamate (KEU) PSMA-targeting moiety and the remaining terminal primary amines were capped with butane-1,2-diol. Non-targeted control dendrimer-drug conjugate (Ctrl-DDC) was formulated without conjugation of KEU. PT-DDC and Ctrl-DDC were characterized using high-performance liquid chromatography, matrix assisted laser desorption ionization mass spectrometry and dynamic light scattering. In vitro and in vivo evaluation of PT-DDC and Ctrl-DDC were carried out in isogenic human prostate cancer PSMA PC3 PIP and PSMA PC3 flu cell lines, and in mice bearing the corresponding xenografts.
RESULTS
PT-DDC was stable in 1×PBS and human blood plasma and required glutathione for DM1 release. Optical, PET/CT and biodistribution studies confirmed the in vivo PSMA-specificity of PT-DDC. PT-DDC demonstrated dose-dependent accumulation and cytotoxicity in PSMA PC3 PIP cells, and also showed growth inhibition of the corresponding tumors. PT-DDC did not accumulate in PSMA PC3 flu tumors and did not inhibit their growth. Ctrl-DDC did not show PSMA specificity.
CONCLUSION
In this study, we synthesized a multimodal theranostic agent capable of delivering DM1 and a radionuclide to PSMA tumors. This approach holds promise for enhancing image-guided treatment of aggressive, metastatic subtypes of prostate cancer.
Topics: Dendrimers; Male; Humans; Glutamate Carboxypeptidase II; Prostatic Neoplasms; Antigens, Surface; Cell Line, Tumor; Animals; Mice; Positron Emission Tomography Computed Tomography; Drug Delivery Systems
PubMed: 38832336
DOI: 10.2147/IJN.S454128 -
Archives of Pathology & Laboratory... Jun 2024The identification of autoantibodies associated with autoimmune liver disease (ALD) is crucial for diagnosis and management. Various laboratory methods have been...
CONTEXT.—
The identification of autoantibodies associated with autoimmune liver disease (ALD) is crucial for diagnosis and management. Various laboratory methods have been introduced to detect autoantibody profiles. However, the variable performance of these assays may create challenges for clinicians and patients.
OBJECTIVE.—
To investigate the concordance rates and diagnostic performance of 2 commercially available assays, line immunoassay (LIA) and digital liquid chip method (DLCM), in patients with ALD.
DESIGN.—
A total of 291 serum samples were collected, consisting of 180 sera from patients with ALD and 111 sera from controls. The samples were detected through LIA and DLCM. The agreement and diagnostic performance of each assay were analyzed.
RESULTS.—
There was substantial to almost perfect agreement among prevalent autoantibodies (anti-mitochondrial antibody M2, antibodies against gp210, Sp100, and Ro52). Nevertheless, the Cohen κ coefficient of some uncommon autoantibodies (anti-LKM-1, anti-LC-1, and anti-SLA/LP) between the 2 methods was not ideal. LIA showed slightly better sensitivity, accuracy, and negative predictive value, while DLCM exhibited slightly higher specificity and positive predictive value.
CONCLUSIONS.—
LIA and DLCM demonstrated comparable performance for the detection of common ALD-related autoantibodies. LIA seemed to be more sensitive, while DLCM displayed more specificity. However, standardization of ALD autoantibody detection still faces challenges between these diverse detection systems. Comprehensive interlaboratory validation is essential to mitigate potential misunderstanding and confusion among patients and clinicians.
PubMed: 38830630
DOI: 10.5858/arpa.2024-0057-OA -
BioRxiv : the Preprint Server For... May 2024Targeted radionuclide therapy is based on injections of cancer-specific molecules conjugated with radioactive nuclides. Despite the specificity of this treatment, it is...
Targeted radionuclide therapy is based on injections of cancer-specific molecules conjugated with radioactive nuclides. Despite the specificity of this treatment, it is not devoid of side-effects limiting its use and is especially harmful for rapidly proliferating organs well perfused by blood, like bone marrow. Optimization of radioconjugates administration accounting for toxicity constraints can increase treatment efficacy. Based on our experiments on disseminated multiple myeloma mouse model treated by Ac-DOTA-daratumumab, we developed a mathematical model which investigation highlighted the following principles for optimization of targeted radionuclide therapy. 1) Nuclide to antibody ratio importance. The density of radioconjugates on cancer cells determines the density of radiation energy deposited in them. Low labeling ratio as well as accumulation of unlabeled antibodies and antibodies attached to decay products in the bloodstream can mitigate cancer radiation damage due to excessive occupation of specific receptors by antibodies devoid of radioactive nuclides. 2) Cancer binding capacity-based dosing. The rate of binding of drug to cancer cells depends on the total number of their specific receptors, which therefore can be estimated from the pharmacokinetic curve of diagnostic radioconjugates. Injection of doses significantly exceeding cancer binding capacity should be avoided since radioconjugates remaining in the bloodstream have negligible efficacy to toxicity ratio. 3) Particle range-guided multi-dosing. The use of short-range particle emitters and high-affinity antibodies allows for robust treatment optimization via initial saturation of cancer binding capacity, enabling redistribution of further injected radioconjugates and deposited dose towards still viable cells that continue expressing specific receptors.
PubMed: 38826403
DOI: 10.1101/2024.05.22.595377 -
Veterinary Clinical Pathology Jun 2024Regenerating island-derived proteins (REG) are upregulated in people with sepsis, pancreatitis, and gastrointestinal diseases. One member of the REG family, namely...
BACKGROUND
Regenerating island-derived proteins (REG) are upregulated in people with sepsis, pancreatitis, and gastrointestinal diseases. One member of the REG family, namely REG3E, was recently identified in pancreatic tissue and plasma of dogs, with high expression in pancreatitis and sepsis.
OBJECTIVES
We aimed to develop and validate an ELISA to measure REG3E concentrations in canine blood.
METHODS
An indirect sandwich ELISA was developed using recombinant canine REG3E protein and polyclonal anti-canine REG3E antibodies raised in guinea pigs and rabbits. Antibody specificity was assessed using western blot and mass spectrometric analysis of protein purified from canine plasma. Assay validation included evaluation of dilutional linearity, parallelism, spiking recovery, repeatability and reproducibility, stability, interferences, and comparison of serum and heparinized plasma.
RESULTS
Antibodies bound specifically to REG3E with no evidence of cross-reactivity with other proteins. The limit of detection of the ELISA was 15 ng/mL, and the lower limit of quantification was 30 ng/mL. The assay demonstrated good to excellent linearity, dilutional and mixing parallelism, and recovery, with mean observed-to-expected ratios of 104%, 107%, 102%, and 92%, respectively, and no evidence of a hook effect. Coefficients of variation were ≤8.5% for repeatability and ≤14.3% for reproducibility at three different levels. Measurements of REG3E in plasma were not significantly influenced by different storage conditions, freeze-thawing cycles, hemolysis, lipemia, or icterus. There was no significant difference between REG3E concentrations in heparinized plasma and serum samples.
CONCLUSIONS
The canine REG3E ELISA has acceptable precision, accuracy, linearity, and reproducibility for the measurement of REG3E in canine plasma and serum.
Topics: Animals; Dogs; Enzyme-Linked Immunosorbent Assay; Reproducibility of Results; Rabbits; Pancreatitis-Associated Proteins; Recombinant Proteins
PubMed: 38825585
DOI: 10.1111/vcp.13352 -
Transfusion and Apheresis Science :... May 2024Limited evidence exists on the distribution of ABO RhD blood groups and prevalence and specificity of red blood cell (RBC) alloantibodies in Aboriginal and Torres Strait...
INTRODUCTION
Limited evidence exists on the distribution of ABO RhD blood groups and prevalence and specificity of red blood cell (RBC) alloantibodies in Aboriginal and Torres Strait Islander peoples of Australia. We investigated RBC alloantibody prevalence and ABO RhD groups in Aboriginal patients undergoing cardiac surgery at a South Australian (SA) tertiary hospital, a major cardiac surgical referral centre for Northern Territory (NT) patients METHODS: Retrospective analysis of all consecutive patients undergoing cardiac surgery at Flinders Medical Centre (FMC) between January 2014 and June 2019. ABO and RhD blood groups, and RBC alloantibody prevalence, specificity, and clinical significance in Aboriginal and non-Aboriginal cardiac patients were determined at time of surgery and on follow up to 2021.
RESULTS
2327 patients were included, 588 (25.3 %) were from NT, and 420 (18.0 %) were Aboriginal. Aboriginal patients had a higher prevalence of ABO group O (59.8 % vs 43.9 %) and RhD positive (99.0 % vs 83.8 %). One-hundred-and-eleven patients had 154 RBC alloantibodies, 57/420 (13.6 %) Aboriginal versus 54/1907 (2.8 %) non-Aboriginal (p < 0.0001). There were higher numbers of IgM alloantibodies in Aboriginal patients (59/77, 76.6 %), with Lewis, P1 and M more common. Sixty patients had antibodies detected at time of surgery, 14 NT patients with previously detected alloantibodies, prior to surgery, presented with a negative antibody screen and 37 had new antibodies detected after cardiac surgery.
CONCLUSION
A high prevalence of IgM alloantibodies was found in Aboriginal compared to non-Aboriginal cardiac surgery patients. The clinical significance of these IgM alloantibodies in Aboriginal peoples requires further investigation.
PubMed: 38823359
DOI: 10.1016/j.transci.2024.103957 -
Applied Microbiology and Biotechnology Jun 2024Getah virus (GETV) is a re-emerging mosquito-borne alphavirus that is highly pathogenic, mainly to pigs and horses. There are no vaccines or treatments available for...
Getah virus (GETV) is a re-emerging mosquito-borne alphavirus that is highly pathogenic, mainly to pigs and horses. There are no vaccines or treatments available for GETV in swine in China. Therefore, the development of a simple, rapid, specific, and sensitive serological assay for GETV antibodies is essential for the prevention and control of GETV. Current antibody monitoring methods are time-consuming, expensive, and dependent on specialized instrumentation, and these features are not conducive to rapid detection in clinical samples. To address these problem, we developed immunochromatographic test strips (ICTS) using eukaryotically expressed soluble recombinant p62-E1 protein of GETV as a labelled antigen, which has good detection sensitivity and no cross-reactivity with other common porcine virus-positive sera. The ICTS is highly compatible with IFA and ELISA and can be stored for 1 month at 37 °C and for at least 3 months at room temperature. Hence, p62-E1-based ICTS is a rapid, accurate, and convenient method for rapid on-site detection of GETV antibodies. KEY POINTS: • We established a rapid antibody detection method that can monitor GETV infection • We developed colloidal gold test strips with high sensitivity and specificity • The development of colloidal gold test strips will aid in the field serologic detection of GETV.
Topics: Animals; Gold Colloid; Antibodies, Viral; Alphavirus; Swine; Sensitivity and Specificity; Chromatography, Affinity; Alphavirus Infections; Swine Diseases; Reagent Strips; China; Enzyme-Linked Immunosorbent Assay
PubMed: 38822832
DOI: 10.1007/s00253-024-13168-5 -
Biomedicine & Pharmacotherapy =... Jul 2024Considering the limited efficacy of current therapies in lung, colorectal, and pancreatic cancers, innovative combination treatments with diverse mechanisms of action...
Considering the limited efficacy of current therapies in lung, colorectal, and pancreatic cancers, innovative combination treatments with diverse mechanisms of action are needed to improve patients' outcomes. Chitinase-3 like-1 protein (CHI3L1) emerges as a versatile factor with significant implications in various diseases, particularly cancers, fostering an immunosuppressive tumor microenvironment for cancer progression. Therefore, pre-clinical validation is imperative to fully realize its potential in cancer treatment. We developed phage display-derived fully human monoclonal CHI3L1 neutralizing antibodies (nAbs) and verified the nAbs-antigen binding affinity and specificity in lung, pancreatic and colorectal cancer cell lines. Tumor growth signals, proliferation and migration ability were all reduced by CHI3L1 nAbs in vitro. Orthotopic or subcutaneous tumor mice model and humanized mouse model were established for characterizing the anti-tumor properties of two CHI3L1 nAb leads. Importantly, CHI3L1 nAbs not only inhibited tumor growth but also mitigated fibrosis, angiogenesis, and restored immunostimulatory functions of immune cells in pancreatic, lung, and colorectal tumor mice models. Mechanistically, CHI3L1 nAbs directly suppressed the activation of pancreatic stellate cells and the transformation of macrophages into myofibroblasts, thereby attenuating fibrosis. These findings strongly support the therapeutic potential of CHI3L1 nAbs in overcoming clinical challenges, including the failure of gemcitabine in pancreatic cancer.
Topics: Animals; Chitinase-3-Like Protein 1; Humans; Colorectal Neoplasms; Pancreatic Neoplasms; Neovascularization, Pathologic; Mice; Cell Line, Tumor; Lung Neoplasms; Cell Proliferation; Fibrosis; Antibodies, Monoclonal; Tumor Microenvironment; Xenograft Model Antitumor Assays; Antibodies, Neutralizing; Antineoplastic Agents, Immunological; Angiogenesis
PubMed: 38820971
DOI: 10.1016/j.biopha.2024.116825 -
PloS One 2024Highly variable pandemic coronavirus SARS-CoV-2, which causes the hazardous COVID-19 infection, has been persistent in the human population since late 2019. A prompt...
Highly variable pandemic coronavirus SARS-CoV-2, which causes the hazardous COVID-19 infection, has been persistent in the human population since late 2019. A prompt assessment of individual and herd immunity against the infection can be accomplished by using rapid tests to determine antiviral antibody levels. The microneutralization assay (MN) is one of the most widely used diagnostic methods that has been proposed to assess the qualitative and quantitative characteristics of virus-specific humoral immunity in COVID-19 convalescents or vaccine recipients. However, some aspects of the assay, such as sensitivity and time cost, need improvement. Here, we developed an express test, which may be potentially used in clinical practice for the assessment of serum-caused SARS-CoV-2 inhibition in infected cell cultures. It implies the detection and counting of coronaviral fluorescent-forming units (FFU) and includes two sequentially used developing components: biotinylated mouse monoclonal antibodies against the recombinant N protein of SARS-CoV-2 (B.1) and the recombinant EGFP-streptavidin fusion protein. Due to the universal specificity of the antibodies, our analytical tool is suitable for the detection of various strains of SARS-CoV-2 when determining both the infectious titer of viruses and the titer of serum virus-neutralizing antibodies. The developed two-component test system is characterized by high sensitivity, a reduced number of analytic stages and low assay cost, as well as by flexibility, since it may be modified for detection of other pathogens using the appropriate antibodies.
Topics: SARS-CoV-2; Humans; COVID-19; Animals; Antibodies, Viral; Vero Cells; Chlorocebus aethiops; Mice; Fluorescent Antibody Technique; Antibodies, Monoclonal; Antibodies, Neutralizing
PubMed: 38820303
DOI: 10.1371/journal.pone.0304534