Clinical predictors of donor antibody titre and correlation with recipient antibody response in a COVID-19 convalescent plasma clinical trial.

BACKGROUND: Convalescent plasma therapy for COVID-19 relies on transfer of anti-viral antibody from donors to recipients via plasma transfusion. The relationship between clinical characteristics and antibody response to COVID-19 is not well defined. We investigated predictors of convalescent antibody production and quantified recipient antibody response in a convalescent plasma therapy clinical trial. METHODS: Multivariable analysis of clinical and serological parameters in 103 confirmed COVID-19 convalescent plasma donors 28 days or more following symptom resolution was performed. Mixed-effects regression models with piecewise linear trends were used to characterize serial antibody responses in 10 convalescent plasma recipients with severe COVID-19. RESULTS: Donor antibody titres ranged from 0 to 1 : 3892 (anti-receptor binding domain (RBD)) and 0 to 1 : 3289 (anti-spike). Higher anti-RBD and anti-spike titres were associated with increased age, hospitalization for COVID-19, fever and absence of myalgia (all P < 0.05). Fatigue was significantly associated with anti-RBD (P = 0.03). In pairwise comparison amongst ABO blood types, AB donors had higher anti-RBD and anti-spike than O donors (P < 0.05). No toxicity was associated with plasma transfusion. Non-ECMO recipient anti-RBD antibody titre increased on average 31% per day during the first three days post-transfusion (P = 0.01) and anti-spike antibody titre by 40.3% (P = 0.02). CONCLUSION: Advanced age, fever, absence of myalgia, fatigue, blood type and hospitalization were associated with higher convalescent antibody titre to COVID-19. Despite variability in donor titre, 80% of convalescent plasma recipients showed significant increase in antibody levels post-transfusion. A more complete understanding of the dose-response effect of plasma transfusion amongst COVID-19-infected patients is needed.

Hemagglutinin stalk-based universal vaccine constructs protect against group 2 influenza A viruses.

Current influenza virus vaccines contain H1N1 (phylogenetic group 1 hemagglutinin), H3N2 (phylogenetic group 2 hemagglutinin), and influenza B virus components. These vaccines induce good protection against closely matched strains by predominantly eliciting antibodies against the membrane distal globular head domain of their respective viral hemagglutinins. This domain, however, undergoes rapid antigenic drift, allowing the virus to escape neutralizing antibody responses. The membrane proximal stalk domain of the hemagglutinin is much more conserved compared to the head domain. In recent years, a growing collection of antibodies that neutralize a broad range of influenza virus strains and subtypes by binding to this domain has been isolated. Here, we demonstrate that a vaccination strategy based on the stalk domain of the H3 hemagglutinin (group 2) induces in mice broadly neutralizing anti-stalk antibodies that are highly cross-reactive to heterologous H3, H10, H14, H15, and H7 (derived from the novel Chinese H7N9 virus) hemagglutinins. Furthermore, we demonstrate that these antibodies confer broad protection against influenza viruses expressing various group 2 hemagglutinins, including an H7 subtype. Through passive transfer experiments, we show that the protection is mediated mainly by neutralizing antibodies against the stalk domain. Our data suggest that, in mice, a vaccine strategy based on the hemagglutinin stalk domain can protect against viruses expressing divergent group 2 hemagglutinins.

Receptor revision of immunoglobulin heavy chain variable region genes in normal human B lymphocytes.

Contrary to the general precepts of the clonal selection theory, several recent studies have provided evidence for the secondary rearrangement of immunoglobulin (Ig) genes in peripheral lymphoid tissues. These analyses typically used transgenic mouse models and have only detected secondary recombination of Ig light chain genes. Although Ig heavy chain variable region (V(H)) genes encode a substantial element of antibody combining site specificity, there is scant evidence for V(H) gene rearrangement in the periphery, leaving the physiological importance of peripheral recombination questionable. The extensive somatic mutations and clonality of the IgD(+)Strictly-IgM(-)CD38(+) human tonsillar B cell subpopulation have now allowed detection of the first clear examples of receptor revision of human V(H) genes. The revised VDJ genes contain "hybrid" V(H) gene segments consisting of portions from two separate germline V(H) genes, a phenomenon previously only detected due to the pressures of a transgenic system.

Somatic hypermutation introduces insertions and deletions into immunoglobulin V genes.

During a germinal center reaction, random mutations are introduced into immunoglobulin V genes to increase the affinity of antibody molecules and to further diversify the B cell repertoire. Antigen-directed selection of B cell clones that generate high affinity surface Ig results in the affinity maturation of the antibody response. The mutations of Ig genes are typically basepair substitutions, although DNA insertions and deletions have been reported to occur at a low frequency. In this study, we describe five insertion and four deletion events in otherwise somatically mutated VH gene cDNA molecules. Two of these insertions and all four deletions were obtained through the sequencing of 395 cDNA clones (approximately 110,000 nucleotides) from CD38+IgD- germinal center, and CD38-IgD- memory B cell populations from a single human tonsil. No germline genes that could have encoded these six cDNA clones were found after an extensive characterization of the genomic VH4 repertoire of the tonsil donor. These six insertions or deletions and three additional insertion events isolated from other sources occurred as triplets or multiples thereof, leaving the transcripts in frame. Additionally, 8 of 9 of these events occurred in the CDR1 or CDR2, following a pattern consistent with selection, and making it unlikely that these events were artifacts of the experimental system. The lack of similar instances in unmutated IgD+CD38- follicular mantle cDNA clones statistically associates these events to the somatic hypermutation process (P = 0.014). Close scrutiny of the 9 insertion/deletion events reported here, and of 25 additional insertions or deletions collected from the literature, suggest that secondary structural elements in the DNA sequences capable of producing loop intermediates may be a prerequisite in most instances. Furthermore, these events most frequently involve sequence motifs resembling known intrinsic hotspots of somatic hypermutation. These insertion/deletion events are consistent with models of somatic hypermutation involving an unstable polymerase enzyme complex lacking proofreading capabilities, and suggest a downregulation or alteration of DNA repair at the V locus during the hypermutation process.

Aspects of B-cell non-Hodgkin's lymphoma development: a transition from immune-reactivity to malignancy.

The development of B-cell lymphomas is an intricate interplay among various pathogenic factors, leading to a multi-step process, encompassing various stages of B-cell maturation. Besides genetic abnormalities, a variety of environmental and microbial factors, as well as disproportional immune-regulatory processes lead to the malignant transformation. Yet, little is known about the exact chain of events, which lead from the physiological polyclonal B-cell activation as a response to exogenous antigens through oligoclonality to a monoclonal, uncontrolled, malignant B-cell proliferation. The aim of the present review was to summarize the potential harmful steps in the development of B-cell lymphomas, according to conventional and novel theories, and to depict therapeutic regimens presently in use as well as to envision future drug developments, beneficial in the battle against this lymphoid malignancy.

Extrarenal Wilms' tumor: staging, treatment, and prognosis.

Extrarenal Wilms' tumors are rare and have mainly been the subject of isolated case reports. This retrospective evaluation of 34 patients suggests a clinical course very similar to that of renal Wilms' tumor. While clinical presentation varies according to the extrarenal localization, investigations to determine the size of the primary tumor (T), regional lymph node involvement (N), and the occurrence of distant metastasis (M) are very similar, ie, chest x-ray, abdominal ultrasound (US), and computed axial tomographic (CAT) scan of the abdomen and chest. Stage grouping according to the pathologic TNM classification showed stage I in 30%, stage II in 10%, stage IIIa in 34%, stage IIIb in 23%, and stage IVa in 3%; four patients could not be staged. Evaluation of therapy and survival indicate the need for postoperative chemotherapy (CT) to all patients, while the same data suggest that the drugs used for renal Wilms' tumor are equally effective for extrarenal Wilms' tumor. Radiotherapy (RT) probably should be reserved for those patients with unresectable gross residual disease and for metastatic disease. The radiation dose used in the reviewed cases varied from 2,000 to 5,000 cGy. However, recent experience suggests that high doses are not justified in renal Wilms' tumor. The estimated overall 2-year survival of the 34 patients is 82% (95% confidence interval, 63% to 92%).

Cleaning-induced arsenic mobilization and chromium oxidation from CCA-wood deck: Potential risk to children.

Concern about children's exposure to arsenic (As) from wood treated with chromated-copper-arsenate (CCA) led to its withdrawal from residential use in 2004. However, due to its effectiveness, millions of American homes still have CCA-wood decks on which children play. This study evaluated the effects of three deck-cleaning methods on formation of dislodgeable As and hexavalent chromium (CrVI) on CCA-wood surfaces and in leachate. Initial wipes from CCA-wood wetted with water showed 3-4 times more dislodgeable As than on dry wood. After cleaning with a bleach solution, 9.8-40.3µg/100cm(2) of CrVI was found on the wood surface, with up to 170µg/L CrVI in the leachate. Depending on the cleaning method, 699-2473mg of As would be released into the environment from cleaning a 18.6-m(2)-deck. Estimated As doses in children aged 1-6 after 1h of playing on a wet CCA-wood deck were 0.25-0.41µg/kg. This is the first study to identify increased dislodgeable As on wet CCA-wood and to evaluate dislodgeable CrVI after bleach application. Our data suggest that As and CrVI in 25-year old CCA-wood still show exposure risks for children and potential for soil contamination.

Plasma cholinesterase activity as an environmental impact biomarker in juvenile green turtles (Chelonia mydas) / Atividade da colinesterase plasmática como biomarcador de impacto ambiental em tartarugas verdes juvenis (Chelonia mydas)

The objective of this study was to evaluate the enzymatic activity of plasma cholinesterase in Chelonia mydas marine turtles belonging to two populations, according to their capture sites, under the absence and probable influence of anthropic effects. A total of 74 animals were used and later divided into two groups, based on the capture site. Blood samples were collected from all captured animals, which were then released into the sea at the site of capture. A descriptive statistical analysis of the plasma cholinesterase activity values and an analysis comparing these values based on the capture site were performed. Samples of heparinized plasma from animals captured at the two different sites were analyzed. Plasma cholinesterase activity ranged from 121 to 248U/L, with a mean and standard deviation of 186.1±30.68U/L. When comparing plasma cholinesterase activity values in individuals based on the capture site, a significant difference was observed. Establishing reference values for different sea turtle populations is necessary to interpret future sampling results and to allow sea turtles to be used as sentinels of ecosystem health. Future studies are needed to evaluate other populations and the activity of plasma cholinesterase in juvenile marine turtles, in relation to environmental contamination.(AU)

O objetivo desse estudo foi avaliar a atividade enzimática da colinesterase plasmática em tartarugas marinhas da espécie Chelonia mydas em duas populações de acordo com o local de captura, sob ausência e provável influência de efeito antrópico. Foi utilizado um total de 74 animais e posteriormente divididos em dois grupos de acordo com o local de captura. Foram coletadas amostras de sangue de todos os animais capturados e em seguida liberados ao mar no mesmo local. Foi realizada uma análise estatística descritiva dos valores da atividade plasmática de colinesterase do total de animais e análise comparando os valores de acordo com o local de captura. Foram analisadas amostras de plasma heparinizado de animais capturados em dois locais distintos. Os valores da atividade plasmática de colinesterase variaram de 121 a 248U/L, com média e desvio padrão de 186.1±30.7U/L. Quando comparados os valores de atividade plasmática da colinesterase nos indivíduos de acordo com o local de captura, foi observada diferença significativa. O estabelecimento de valores de referência para diferentes populações de tartarugas marinhas são necessários para interpretar os futuros resultados amostrais e permitir que as tartarugas marinhas sejam usadas como sentinelas da saúde do ecossistema. Estudos futuros são necessários para avaliar outras populações e a atividade da colinesterase plasmática de tartarugas marinhas juvenis em relação à contaminação ambiental.(AU)

Intestinal transport of zinc and folic acid: a mutual inhibitory effect.

Recent observations suggest an inverse relationship between folic acid intake and zinc nutriture and indicate an interaction between folic acid and zinc at the intestinal level. To define that interaction, we designed in vivo and in vitro transport studies in which folic acid transport in the presence of zinc, as well as zinc transport in the presence of folic acid was examined. These studies show that zinc transport is significantly decreased when folate is present in the intestinal lumen. Similarly folic acid transport is significantly decreased with the presence of zinc. To determine whether this intestinal inhibition is secondary to zinc and folate-forming complexes, charcoal-binding studies were performed. These studies indicate that zinc and folate from complexes at pH 2.0, but that at pH 6.0, these complexes dissolve. Therefore, our studies suggest that under normal physiological conditions a mutual inhibition between folate and zinc exists at the site of intestinal transport.

Assessment of ovulation by ultrasound and plasma estradiol determinations.

The timing of ovulation was studied daily during the periovular period in 19 women attending an artificial insemination clinic during 24 menstrual cycles; B-mode ultrasound examination and plasma estradiol (E2) determinations were used. The maximum diameters of the ovarian follicles and the peak plasma E2 determinations all occurred within 2 days prior to and including the day of ovulation, as determined by conventional means, in the 12 normal cycles studied. The average values on the day prior to ovulation (day -1) for maximum follicle size and peak E2 determination were 2.5 cm and 1660 pmole/liter, respectively. Two definite ultrasonic patterns were noted after ovulation: The follicle either disappeared or filled with internal echoes. It is concluded from the preliminary study that ultrasound examination and plasma E2 determinations are equally effective in predicting the time of ovulation.

Automated water-path ultrasonic examination of scrotum.

A new method of examining the scrotum by ultrasound with an automated water-path scanner is described. This method allows immobility of the scrotum in a more acceptable and painless way than in contact scanning methods. Panoramic scans of the entire scrotum and precision tomographic plane placement are advantages over conventional techniques. A description of normal scrotal anatomy includes the mediastinum of the testis, an important scrotal landmark. Representative cases of benign and malignant scrotal disease are presented to emphasize the utility of water-path scanning.

Early evaluation of coronary artery bypass grafts: CT or selective angiography.

A prospective study was performed in 27 patients to compare the value of computed tomography and selective angiography in assessing coronary artery graft patency in the early post-operative period. The sensitivity of CT to graft patency was 85% with no falsely patent determinations. Dynamic CT was not found useful in predicting graft stenosis. There were no complications associated with CT studies, and two related to selective angiography. It is concluded that CT is the procedure of choice for graft evaluation in the early post-operative period; but that angiography is mandatory for the assessment of late symptom recurrence. A review is made of the results described in previous series.

The management of advanced thyroid cancer.

Although the prognosis of the majority of patients with thyroid cancer is excellent there are some patients with advanced thyroid cancer in whom the prognosis is poor. Such patients include: patients with locally invasive or recurrent differentiated thyroid cancer and medullary thyroid cancer, and all patients with anaplastic cancer. The management of patients with advanced thyroid cancer is reviewed with emphasis on the role of external beam radiotherapy.

Metalaxyl toxicity, uptake, and distribution in several ornamental plant species.

Phytoremediation depends on the ability of plants to tolerate and assimilate contaminants. This research characterized the interaction between several ornamental plant species and the fungicidal active ingredient, metalaxyl [N-(2,6-dimethylphenyl)-N-(methoxyacetyl)alanine methyl ester]. Species evaluated included sweetflag (Acorus gramineus Sol. ex Aiton), canna (Canna hybrida L. 'Yellow King Humbert'), parrotfeather [Myriophyllum aquaticum (Vell.) Verdc.], and pickerelweed (Pontederia cordata L.). Metalaxyl tolerance levels for each species were determined by exposing plants for 7 d to solutions containing 0, 5, 10, 25, 50, 75, or 100 mg metalaxyl L-1 aqueous nutrient media. Response endpoints included fresh mass production after 7 d exposure and 7 d post-exposure and quantum efficiency using dark-adapted (Fv/Fm) and light-adapted (fluorescence yields) plants. Metalaxyl uptake and distribution within the plant was determined by growing plants in aqueous nutrient media containing 1.18 x 10(6) Bq L-1 [14C]metalaxyl (0.909 mg L-1) for 1, 3, 5, or 7 d. Plant tissues were combusted and analyzed by liquid scintillation counting. Metalaxyl had no effects on the endpoints measured, except for fresh mass production of sweetflag at the 75 and 100 mg L-1 treatment levels. However, leaf necrosis was apparent in most species after 5 d exposure to concentrations greater than 25 mg L-1. Metalaxyl removal from the spiked nutrient media ranged from 15 to 60% during the 7-d exposure period. The majority of metalaxyl removed from the solution was detected within individual plants. In nearly all cases, activity from the radiolabeled pesticide accumulated in the leaves. Uptake of metalaxyl was correlated with water uptake throughout the 7 d. These results suggest that all species examined may be good candidates for incorporation into a phytoremediation scheme for metalaxyl.

Diversity of the Ig repertoire is maintained with age in spite of reduced germinal centre cells in human tonsil lymphoid tissue.

Humans and almost all species studied to date exhibit a decreased responsiveness to immunization and increased autoimmunity with age. While this has been observed clinically for decades, only recently has an understanding of the molecular basis for these changes begun to be appreciated. Studies of the B-cell aspects of these changes in ageing mice and the very few reports in ageing humans have not been conclusive. Here we examine the nucleotide sequence of over 1250 VH transcripts from the tonsils of individuals of various ages for changes to the VH4 immunoglobulin repertoire. An exhaustive examination of VH, DH and JH gene segment utilization revealed a remarkable similarity of the repertoires. The extent of somatic hypermutation was fully maintained or even increased by some measures into the eighth decade of life. However, we found by middle age that the representation of naïve and germinal centre B-cell subpopulations changed relative to total B lymphocytes in the tonsil. While the percentage of naïve and germinal centre B-cell subpopulations changes during the second half of life, these findings suggest that even with advancing age, humans remain capable of generating an extremely diverse Ig repertoire while maintaining a similar spectrum of Ig rearrangements once the germinal centre reaction begins.

A genome-scale assessment of peripheral blood B-cell molecular homeostasis in patients with rheumatoid arthritis.

OBJECTIVE: While rheumatoid arthritis (RA) is considered a prototypical autoimmune disease, the specific roles of B-cells in RA pathogenesis is not fully delineated. METHODS: We performed microarray expression profiling of peripheral blood B-cells from RA patients and controls. Data were analysed using differential gene expression analysis and 'gene networking' analysis (characterizing clusters of functionally inter-relelated genes) to identify both regulatory genes and the pathways in which they participate. Results were confirmed by quantitative real-time polymerase chain reaction and by measuring the levels of 10 serum cytokines involved in the pathways identified. RESULTS: Genes regulating and effecting the cell-cycle, proliferation, apoptosis, autoimmunity, cytokine networks, angiogenesis and neuro-immune regulation were differentially expressed in RA B-cells. Moreover, the serum levels of several soluble factors that modulate these pathways, including IL-1beta, IL-5, IL-6, IL-10, IL-12p40, IL-17 and VEGF were significantly increased in this cohort of RA patients. CONCLUSIONS: These results outline aspects of the multifaceted role B-cells play in RA pathogenesis in which immune dysregulation in RA modulates B-cell biology and thereby contributes to the induction and perpetuation of a pathogenic humoral immune response.

Successful birth after previous tubal ectopic pregnancies.

A case, in which tubal conservation in an ectopic pregnancy resulted in the birth of a living infant, is described. It is hoped that tubal conservation in ectopic pregnancies, which was previously thought to be unwise, will be considered in future.

PIP: Tubal conservation following ectopic pregnancy resulted in a successful birth in this case report of a 25-year-old woman who in 1978 underwent a laparotomy which revealed an ectopic pregnancy of the ampulla of the right tube, which had ruptured. The pregnancy was evacuated, and conservative operative repair of the tube was made to prepare for later microsurgery. In 1978, 7 months later and before microsurgery, the patient presented with a 6-week intrauterine pregnancy, the course and outcome of which were normal and uneventful. It is hoped that tubal conservation in ectopic pregnancies, which was previously thought unwise, will be considered in the future.