Bone Marrow Langerhans Cell Histiocytosis in Association with Kasabach-Merritt Syndrome: The Difficulty of a Differential Diagnosis

Langerhans cell histiocytosis is a rare haematological disorder with variable clinical findings and a high mortality rate. On the other hand, Kasabach-Merritt syndrome is of rare onset at adult age, requiring the simultaneous presentation of vascular lesion, thrombocytopenia, and consumptive coagulopathy. We present the first reported case of both diseases in a single patient and highlight the difficulties of diagnostic. A 69-year-old woman with immune thrombocytopenic purpura underwent surgery for the removal of giant skin haemangiomas. During post-operative care, intravascular disseminated coagulopathy developed. After weeks of corticosteroids and immunosuppressive therapy with no clinical improvement, pulmonary tuberculosis was diagnosed and appropriate treatment initiated. Despite all the efforts, the patient's clinical condition kept worsening and she eventually died. An autopsy revealed bone marrow Langerhans cell histiocytosis. In this case, the patient's autoimmune background together with tuberculosis and intravascular disseminated coagulopathy masked the presentation and made the diagnosis of a rapidly progressive fatal disease very difficult.info:eu-repo/semantics/publishedVersio

Advancing downstream purification of cell and gene therapy medicinal products

The advent of advanced therapies in the biopharmaceutical industry has moved the spotlight into complex products such as viral vectors or stem cells, holding great promise in a myriad of clinical targets. Currently, the challenge for a widespread application of these new biopharmaceuticals is the development of cost-effective bioprocesses while maintaining product\u27s bioactivity and quality attributes. This presentation will focus on the latest advancements on downstream purification of cell and gene therapy medicinal products, supported by process innovation and the flexibility of old, but robust technologies such as tangential flow filtration (TFF) and chromatography. Improvement of purification yields of virus based biopharmaceuticals can be achieved through the rational development of alternative strategies, combining different modes of operation, such as flow through purification or multi-column chromatography, together with the recent developments of chromatographic media and fundamental understanding of the adsorption phenomena as reported for the case of gene therapy medicinal products. Critical quality attributes of cell based medicinal products cannot be compromised by the processing route chosen. The use of the already established TFF technology has the potential to improve the purity of cell based products, with the evaluation of critical process parameters of cell concentration and washing being of paramount importance. The purity of such products can also be incremented with the use of negative mode expanded bed adsorption chromatography with a new multimodal prototype matrix based on core–shell bead technology as demonstrated for the case of human mesenchymal stem cells. In summary, the advancement of the purification of complex biopharmaceutical entities, such as the ones here reported, can be described as an incremental process, but still with space for inn

p518, a small floR plasmid from a South American isolate of Actinobacillus pleuropneumoniae

A small (3.9 kb) plasmid (p518), conferring resistance to florfenicol (MIC >8 μg/mL) and chloramphenicol (MIC >8 μg/mL) was isolated from an Actinobacillus pleuropneumoniae clinical isolate from Southeastern Brazil. To date, this is the smallest florfenicol resistance plasmid isolated from a member of the Pasteurellaceae. The complete nucleotide of this plasmid revealed a unique gene arrangement compared to previously reported florfenicol resistance plasmids found in other members of the Pasteurellaceae. In addition to the floR gene and a lysR gene, common to various florfenicol resistance plasmids, p518 also encodes strA and a partial strB sequence. An origin of replication (oriV) similar to that in the broad host range plasmid, pLS88, was identified in p518, and transformation into Escherichia coli MFDpir confirmed the ability to replicate in other species. Mobilisation genes appear to have been lost, with only a partial mobC sequence remaining, and attempts to transfer p518 from a conjugal donor strain (E. coli MFDpir) were not successful, suggesting this plasmid is not mobilisable. Similarly, attempts to transfer p518 into a competent A. pleuropneumoniae strain, MIDG2331, by natural transformation were also not successful. These results suggest that p518 may be only transferred by vertical descent

The lymphocyte-to-monocyte ratio: An added value for death prediction in heart failure

BACKGROUND AND AIM: Leukocytes and their subpopulation have been long implicated in the progression of the syndrome of heart failure (HF), especially heart infiltration cells. Previous reports have suggested that they can predict worse outcome in patients with HF, and can also affect the function of other cells and myocardial extracellular matrix remodeling process. However, the lymphocyte-to-monocyte ratio (LMR) and its possible value as prognostic marker have not been evaluated. METHODS AND RESULTS: A total of 390 patients with acute HF were recruited and followed for 6 months. Their total blood count with leukocyte differential was obtained. Two groups were formed according to the endpoints of HF death and optimal cut-off value of LMR, and were compared. A multivariate Cox-regression model was used to establish the prognostic value with the endpoints of HF and all-cause mortality. Median age of the patients was 78 years and 48.5% of them were men. No major difference was observed between the clinical characteristics of the two groups. Patients who died of HF had significantly higher values of B-type natriuretic peptide and lower values of LMR. Leukocyte and monocyte counts revealed a multivariate-adjusted risk for both endpoints, whereas relative lymphocyte counts had only significant value for all-cause mortality. The multivariate-adjusted hazard ratios for the 6-month HF and all-cause mortality in patients with LMR values < 2.0 were, respectively, 2.28 (95% CI: 1.25-4.15) and 2.39 (95% CI: 1.39-4.10). CONCLUSION: Our results show that, upon discharge from hospital after an episode of acute HF, a lower value of LMR is independently associated with a higher risk of mortality within 6 months

Serum Levels of Vitamin D and Dental Caries in 7-Year-Old Children in Porto Metropolitan Area

Vitamin D deficiency has been associated with significant changes in dental structures. In children, it can induce enamel and dentin defects, which have been identified as risk factors for caries. This study aimed to assess the association between low serum 25-hydroxyvitamin D (25(OH) D) levels ( 0), and advanced caries (d3–6mft/D3–6MF > 0). Serum 25(OH) D was measured using a competitive electrochemiluminescence immunoassay protein-binding assay. Bivariate analysis and multivariate logistic regression were used. Advanced caries in permanent teeth was significantly associated with children’s vitamin D levels <30 ng/mL, gastrointestinal disorders, higher daily intake of cariogenic food, and having had a dental appointment at ≤7 years old. Optimal childhood levels of vitamin D may be considered an additional preventive measure for dental caries in the permanent dentition.Generation XXI was funded by Programa Operacional de Saúde (Regional Department of Ministry of Health). It has support from the Portuguese Foundation for Science and Technology (FCT) and from the Calouste Gulbenkian Foundation. Ana Cristina Santos holds a FCT Investigator contract info:eu-repo/grantAgreement/FCT/Investigador FCT/IF/01060/2015/CP1319/CT0001/PT. The authors received no specific funding for this work

Establishing the design space of a filtration-based operation for the concentration of human pluripotent stem cells

Stem cell manufacturing is often very challenging due to the complexity of the biological system. Quality-by-design (QbD), a risk-based framework based on relating process parameters and product quality, can cope with such complexity in process design [1] aiding to develop robust and reproducible unit operations. This work describes a shortcut approach for the design of tangential flow filtration for the concentration of human induced pluripotent stem cells (hiPSC), supported by design of experiments (DoE) approach. First, critical quality attributes (CQA), corresponding to the characteristics that ensure the final product quality, and critical process parameters (CPP), which directly affect cells’ CQA, are identified. Thereafter, a design space is developed, studying how a range of variability in CPP allows to achieve CQA [2]. Thus, CPP of shear rate, permeate flux and cell load were considered, and initially their impact on hiPSC recovery yield and viability responses was studied. A full factorial design confirmed significant interaction effects between all CPP, affecting both responses. The developed statistical model predicted that high shear rate (3000 s-1), permeate flux (250 LMH) and medium cell load (2 x 106 cell/cm2) would maximize both cell recovery yield and viability, where over 80% of hiPSC would be recovered after a volume reduction factor of 20 with high viability (over 93%). Such conditions were validated experimentally, and by performing a robustness analysis, the success rate of these operating conditions was assessed (65 - 70%). A parametric study was then conducted, identifying that increasing the shear rate (up to 3370 s-1) allowed to achieve the specified requirements for cell recovery yield (\u3e 80%) and viability (\u3e 90%) in 100% of the cases and no impact in hiPSC’s CQA in terms of identity, proliferation capacity and pluripotency was observed. References [1] Lipsitz YY et al, Nat. Biotechnol. 2016, 34, 393–400. [2] Food and Drug Administration, Guidance for Industry: Q8(R2) pharmaceutical development. The authors acknowledge iNOVA4Health Research Unit (LISBOA-01-0145-FEDER-007344), and Fundação para a Ciência e Tecnologia (FCT, Portugal) for funding the project CARDIOSTEM (MITP-TB/ECE/0013/2013), and the grants SFRH/BD/51940/2012 (MIT-Portugal), SFRH/BD/51573/2011

Education can improve the negative perception of a threatened long-lived scavenging bird, the Andean condor

Human-wildlife conflicts currently represent one of the main conservation problems for wildlife species around the world. Vultures have serious conservation concerns, many of which are related to people's adverse perception about them due to the belief that they prey on livestock. Our aim was to assess local perception and the factors influencing people's perception of the largest scavenging bird in South America, the Andean condor. For this, we interviewed 112 people from Valle Fértil, San Juan province, a rural area of central west Argentina. Overall, people in the area mostly have an elementary education, and their most important activity is livestock rearing. The results showed that, in general, most people perceive the Andean condor as an injurious species and, in fact, some people recognize that they still kill condors. We identified two major factors that affect this perception, the education level of villagers and their relationship with livestock ranching. Our study suggests that conservation of condors and other similar scavengers depends on education programs designed to change the negative perception people have about them. Such programs should be particularly focused on ranchers since they are the ones who have the worst perception of these scavengers. We suggest that highlighting the central ecological role of scavengers and recovering their cultural value would be fundamental to reverse their persecution and their negative perception by people.Fil: Cailly Arnulphi, Verónica Beatríz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Juan. Centro de Investigaciones de la Geosfera y Biosfera. Universidad Nacional de San Juan. Facultad de Ciencias Exactas Físicas y Naturales. Centro de Investigaciones de la Geosfera y Biosfera; ArgentinaFil: Lambertucci, Sergio Agustin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; ArgentinaFil: Borghi, Carlos Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Juan. Centro de Investigaciones de la Geosfera y Biosfera. Universidad Nacional de San Juan. Facultad de Ciencias Exactas Físicas y Naturales. Centro de Investigaciones de la Geosfera y Biosfera; Argentin

Dynamics of a Dual SARS-CoV-2 Lineage Co-Infection on a Prolonged Viral Shedding COVID-19 Case: Insights into Clinical Severity and Disease Duration

A few molecularly proven severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) cases of symptomatic reinfection are currently known worldwide, with a resolved first infection followed by a second infection after a 48 to 142-day intervening period. We report a multiple-component study of a clinically severe and prolonged viral shedding coronavirus disease 2019 (COVID-19) case in a 17-year-old Portuguese female. She had two hospitalizations, a total of 19 RT-PCR tests, mostly positive, and criteria for releasing from home isolation at the end of 97 days. The viral genome was sequenced in seven serial samples and in the diagnostic sample from her infected mother. A human genome-wide array (>900 K) was screened on the seven samples, and in vitro culture was conducted on isolates from three late samples. The patient had co-infection by two SARS-CoV-2 lineages, which were affiliated in distinct clades and diverging by six variants. The 20A lineage was absolute at the diagnosis (shared with the patient's mother), but nine days later, the 20B lineage had 3% frequency, and two months later, the 20B lineage had 100% frequency. The 900 K profiles confirmed the identity of the patient in the serial samples, and they allowed us to infer that she had polygenic risk scores for hospitalization and severe respiratory disease within the normal distributions for a Portuguese population cohort. The early-on dynamic co-infection may have contributed to the severity of COVID-19 in this otherwise healthy young patient, and to her prolonged SARS-CoV-2 shedding profile.The authors acknowledge the support of the i3S Scientific Platforms BioSciences Screening and Genomics, members of the national infrastructure PPBI-Portuguese Platform of Bioimaging (PPBI-POCI-01-0145-FEDER-022122), PT-OPENSCREEN, GenomePT project (POCI-01-0145-FEDER-022184)