Flow cytometric assessment of the reactivity of a panel of monoclonal antibodies (mAb) against two populations of human dendritic cells (DC)

BACKGROUND: The identification of antigens on human DC has been a very difficult and elusive task because of the lack of appropriate reagents. Therefore, we evaluated by flow cytometry a panel of mAb that recognize antigens on human DC, aiming to determine the kinetics of DC antigen expression at 7, 14, 21 and 28 days in (i) Dermal DC like cells (Mo-DC) and (ii) Langerhans cell like DC (Mo-LC). In addition we aimed to identify markers for DC subpopulations. RESULTS: It was found at day 7, that mAb BG6, HP-F1, BU10, RFD-1, CMRF-44 recognized <20% of Mo-DC. In contrast, 7H5, ZM3.8, CDlb/c, 55K-2, MMR1.16, MMR190.BB3 and L25 reacted with >50% of Mo-DC. Moreover, 7H5, ZM3.8, CMRF-56, CDlb/c, 55K-2, MMR1.16, MMR190.BB3 and L25 showed increased MFI reactivity against Mo-DC. mAb BG6, BU10 and CMRF-44 recognized <20% Mo-LC while RFD-1 reacted with 21% of Mo-LC. In contrast, HP-F1 showed 87% of Mo-LC positive. Also, 7H5, ZM3.8, RFD-7, MR15-2, CDlb/c, 55K-2, MMR1.16, MMR190.BB3 and L25 reacted with >50% of Mo-LC. The increase in % of positive cells was paralleled by MFI increases. At day 14, fourteen mAb recognized >50% of the Mo-DC, while five recognized 20-50% of Mo-DC. BG6 reacted with 7% of the Mo-DC. Nineteen mAb recognized >48% of Mo-LC while BG6 had negative reactivity. At day 21 and 28, all mAb reacted with >20% of Mo-DC and yielded a significant MFI with Mo-DC. Also nineteen mAb yielded significant MFI with Mo-LC while RFD-7 did not. CONCLUSIONS: The immunophenotyping assays demonstrated differences between the two DC populations as well as variations in the reactivity of the mAb at diverse time points, suggesting the existence of subpopulations within the Mo-DC and Mo-LC

Review of emerging japanese encephalitis virus: new aspects and concepts about entry into the brain and inter-cellular spreading

Japanese encephalitis virus (JEV) is an emerging flavivirus of the Asia-Pacific region. More than two billion people live in endemic or epidemic areas and are at risk of infection. Recently, the first autochthonous human case was recorded in Africa, and infected birds have been found in Europe. JEV may spread even further to other continents. The first section of this review covers established and new information about the epidemiology of JEV. The subsequent sections focus on the impact of JEV on humans, including the natural course and immunity. Furthermore, new concepts are discussed about JEVs entry into the brain. Finally, interactions of JEV and host cells are covered, as well as how JEV may spread in the body through latently infected immune cells and cell-to-cell transmission of virions or via other infectious material, including JEV genomic RNA

A newly discovered muscle: The tensor of the vastus intermedius

The quadriceps femoris is traditionally described as a muscle group composed of the rectus femoris and the three vasti. However, clinical experience and investigations of anatomical specimens are not consistent with the textbook description. We have found a second tensor-like muscle between the vastus lateralis (VL) and the vastus intermedius (VI), hereafter named the tensor VI (TVI). The aim of this study was to clarify whether this intervening muscle was a variation of the VL or the VI, or a separate head of the extensor apparatus. Twenty-six cadaveric lower limbs were investigated. The architecture of the quadriceps femoris was examined with special attention to innervation and vascularization patterns. All muscle components were traced from origin to insertion and their affiliations were determined. A TVI was found in all dissections. It was supplied by independent muscular and vascular branches of the femoral nerve and lateral circumflex femoral artery. Further distally, the TVI combined with an aponeurosis merging separately into the quadriceps tendon and inserting on the medial aspect of the patella. Four morphological types of TVI were distinguished: Independent-type (11/26), VI-type (6/26), VL-type (5/26), and Common-type (4/26). This study demonstrated that the quadriceps femoris is architecturally different from previous descriptions: there is an additional muscle belly between the VI and VL, which cannot be clearly assigned to the former or the latter. Distal exposure shows that this muscle belly becomes its own aponeurosis, which continues distally as part of the quadriceps tendon

The posterior ridge of the greater tuberosity of the humerus: a suitable landmark for the posterior approach to the shoulder joint?

The purpose of this study was to evaluate the posterior ridge of the greater tuberosity, a palpable prominence during surgery, as a landmark for the posterior approach to the glenohumeral joint.Methods: Twenty-five human cadaveric shoulders were dissected. In 5 cases, a full-thickness rotator cuff tear was present. The posterior surgical anatomy was defined, and the distance from the ridge to the interval between the infraspinatus (IS) and teres minor (TM) muscle, the distance from the ridge to the inferior border of the glenoid (IBG), and the distance between the IS-TM interval and the IBG were determined.Results: In all specimens, a prominent ridge on the posterior greater tuberosity lateral to the articular margin could be identified. The IS-TM interval was located, on average, 3 mm proximal to this ridge. The IS-TM interval corresponded to a point 5 mm proximal to the IBG. In all shoulders, the ridge was located, on average, 8 mm proximal to the IBG. The plane of the IS-TM interval showed a vertically oblique direction.Conclusion: The posterior ridge of the greater tuberosity is a suitable landmark to locate the internervous plane between the IS and TM and should not be crossed distally. Unlike other landmarks, the ridge moves with the humeral head, making it is less dependent on the patient's size, sex, and arm position and the quality of the rotator cuff. The ridge is always located proximal to the insertion of the TM and IBG

Professional Certifications of a University Master’s Degree and Employability: Impact on Students’ Perception

[Abstract]: Professional certifications represent for many university degrees, especially postgraduate degrees, a recognition of their academic quality and the future employability of the graduates. This article contributes to the analysis of the impact of external accreditations on students’ perception of employability and satisfaction. We offer a case of study, a Master of Science (MSc) in Banking and Finance that became the first academic degree in Spain to obtain the two professional accreditations required for employees in financial institutions since 2019. A survey to a sample of students who graduated two academic years before and two years after the MSc was recognised is used to measure students' motivations for enrolment and satisfaction. The results provide significant evidence that professional accreditation became a key motivation for students to enrol the master, is associated with a more diverse geographical origin of students, and students highlight the higher quality and better coordination of the teaching staff.R&D+i Program Oriented to Society’s Challenges ; PID2019-106273RBI0

Human dendritic cells process and present Listeria antigens for in vitro priming of autologous CD4+ T lymphocytes

The role of human dendritic cells (DC) in the immune response toward intracellularly growing Listeria was analyzed under in vitro conditions using several morphological and functional methods. DC incubated with Listeria innocua and L. monocytogenes, respectively, readily phagocytosed the bacteria. Listeria did not impair viability and immunogenic potential of human DC. Listerial antigens were found to be processed within the lysosomal compartment of DC and colocalized with major histocompatibility complex (MHC) class II molecules, as shown by fluorescence and transmission electron microscopy. DC challenged with apathogenic L. innocua were highly effective in priming autologous naïve T cells (mainly CD4+) in vitro. The T cells strongly proliferated in the presence of DC incubated with L. innocua, which could be significantly inhibited by anti-MHC II mAb. L. innocua-primed T cells were also successfully stimulated by DC harboring the pathogenic L. monocytogenes, either the wild-type strain EGD or the p60 reduced mutant strain RIII. From our results, we conclude that human DC infected with nonpathogenic intracellular bacteria are able to efficiently prime naïve T cells, which are then suitable for recognition of antigens derived from related virulent bacterial species. This in vitro human model provides an interesting tool for basic research in infectious immunology and possibly for a new immunotherap

The interaction between the vastus medialis and vastus intermedius and its influence on the extensor apparatus of the knee joint

Although the vastus medialis (VM) is closely associated with the vastus intermedius (VI), there is a lack of data regarding their functional relationship. The purpose of this study was to investigate the anatomical interaction between the VM and VI with regard to their origins, insertions, innervation and function within the extensor apparatus of the knee joint.Methods: Eighteen human cadaveric lower limbs were investigated using macro-dissection techniques. Six limbs were cut transversely in the middle third of the thigh. The mode of origin, insertion and nerve supply of the extensor apparatus of the knee joint were studied. The architecture of the VM and VI was examined in detail, as was their anatomical interaction and connective tissue linkage to the adjacent anatomical structures.Results: The VM originated medially from a broad hammock-like structure. The attachment site of the VM always spanned over a long distance between: (1) patella, (2) rectus femoris tendon and (3) aponeurosis of the VI, with the insertion into the VI being the largest. VM units were inserted twice—once on the anterior and once on the posterior side of the VI. The VI consists of a complex multi- layered structure. The layers of the medial VI aponeurosis fused with the aponeuroses of the tensor vastus intermedius and vastus lateralis. Together, they form the two- layered intermediate layer of the quadriceps tendon. The VM and medial parts of the VI were innervated by the same medial division of the femoral nerve.Conclusion: The VM consists of multiple muscle units inserting into the entire VI. Together, they build a potential functional muscular complex. Therefore, the VM acts as an indirect extensor of the knee joint regulating and adjusting the length of the extensor apparatus throughout the entire range of motion. It is of clinical importance that, besides the VM, substantial parts of the VI directly contribute to the medial pull on the patella and help to maintain medial tracking of the patella during knee extension. The interaction between the VM and VI, with responsibility for the extension of the knee joint and influence on the patellofemoral function, leads readily to an understanding of common clinical problems found at the knee joint as it attempts to meet contradictory demands for both mobility and stability. Surgery or trauma in the anteromedial aspect of the quadriceps muscle group might alter a delicate interplay between the VM and VI. This would affect the extensor apparatus as a whole

A detailed review on the clinical anatomy of the pectoralis major muscle

The pectoralis major is a muscle of the upper limb girdle. This muscle has a unique morphological architectonic and a high rate of clinical applications. However, there is lack of data regarding the morphological and functional interactions of the pectoralis major with other muscle and fascial compartments. According to the applied knowledge, the “Humero-pectoral” morpho-functional concept has been postulated. The purpose of this review was the dissectible investigation of the muscle anatomy and literature review of surgical applications

Difusividade da água no solo: um método simples de laboratório para sua determinação

Soil water diffusivity (D) is an important hydraulic property that is fundamental to characterize unsaturated watertransport. Its determination is complex, time-consuming and requires expensive instruments. The objectives of this workwere: to propose a simple and low-cost laboratory methodology to determine D function; and to analyze the influence of soilmanagement systems on D and Sorptivity (S). The studied soil was classified as a vertic Paleudol. The first 10 cm of the soilunder three different management systems (T1: Natural grassland, T2: direct drilling, and T3: Polyphitic Pasture) was sampled.The samples were sieved and packed into horizontal columns. The columns were analyzed under horizontal infiltration and Dwas determined by variations of water content as a function of time for fixed positions, obtained from low-cost soil moisturecapacitance sensors. The results showed that the proposed methodology is valid in the studied soils. Soil management systemsignificantly affected D and S. They were greater for T2 compared with T1 and T3 (D varied between 0.00033 and 0.0321cm2.s-1). This means that the soil under T2 can transmit water faster under non-saturated conditions as compared with thesoil under grazing. In conclusion, the proposed methodology allowed to determine D in a simple and low-cost way, and todetermine the influence of these properties on productive conditions.A difusividade da água no solo (D) é uma importante propriedade hidráulica para caracterizar o transporte não saturado de água no solo. Essa determinação é complexa devido ao tempo gasto para isso e ao elevado custo dos equipamentos necessários. Os objetivos desse trabalho foram: propor uma metodologia simples e de baixo custo de laboratório para determinar uma função para D e para analisar a influência de sistemas de manejo do solo em D e na sortividade do solo (S). O solo estudado foi classificado como um Paleudol vertico. As amostras deformadas foram coletadas em três sistemas de manejos (T1: Campo Nativo, T2: Plantio Direto e T3: pastagem). Foram retirados os primeiros 10 centímetros do solo. Essas amostras foram peneiradas e depois foram acomodadas em colunas horizontais. As colunas foram submetidas a infiltração horizontal e D foi determinado pela variação do conteúdo de água em função do tempo, com auxílio de sensores de capacitância para a determinação da umidade do solo. Os resultados indicam que a metodologia proposta é válida para os solos desse estudo. O sistema de manejo do solo indicou diferença significativa em D e S. Foram melhores no tratamento T2 comparado com T1 e T3 (D apresentou variação entre 0,00033 e 0,0321 cm2 s-1). Isso indica que o solo na condição T2 conduz água mais rápida na condição não saturada, que o solo manejado com pastejo. Assim, a metodologia proposta permite determinar D em uma amostra e com baixo custo, além de verificar a influência dessas propriedades em condições produtivas.Facultad de Ciencias Agrarias y Forestale

Difusividade da água no solo: um método simples de laboratório para sua determinação

Soil water diffusivity (D) is an important hydraulic property that is fundamental to characterize unsaturated watertransport. Its determination is complex, time-consuming and requires expensive instruments. The objectives of this workwere: to propose a simple and low-cost laboratory methodology to determine D function; and to analyze the influence of soilmanagement systems on D and Sorptivity (S). The studied soil was classified as a vertic Paleudol. The first 10 cm of the soilunder three different management systems (T1: Natural grassland, T2: direct drilling, and T3: Polyphitic Pasture) was sampled.The samples were sieved and packed into horizontal columns. The columns were analyzed under horizontal infiltration and Dwas determined by variations of water content as a function of time for fixed positions, obtained from low-cost soil moisturecapacitance sensors. The results showed that the proposed methodology is valid in the studied soils. Soil management systemsignificantly affected D and S. They were greater for T2 compared with T1 and T3 (D varied between 0.00033 and 0.0321cm2.s-1). This means that the soil under T2 can transmit water faster under non-saturated conditions as compared with thesoil under grazing. In conclusion, the proposed methodology allowed to determine D in a simple and low-cost way, and todetermine the influence of these properties on productive conditions.A difusividade da água no solo (D) é uma importante propriedade hidráulica para caracterizar o transporte não saturado de água no solo. Essa determinação é complexa devido ao tempo gasto para isso e ao elevado custo dos equipamentos necessários. Os objetivos desse trabalho foram: propor uma metodologia simples e de baixo custo de laboratório para determinar uma função para D e para analisar a influência de sistemas de manejo do solo em D e na sortividade do solo (S). O solo estudado foi classificado como um Paleudol vertico. As amostras deformadas foram coletadas em três sistemas de manejos (T1: Campo Nativo, T2: Plantio Direto e T3: pastagem). Foram retirados os primeiros 10 centímetros do solo. Essas amostras foram peneiradas e depois foram acomodadas em colunas horizontais. As colunas foram submetidas a infiltração horizontal e D foi determinado pela variação do conteúdo de água em função do tempo, com auxílio de sensores de capacitância para a determinação da umidade do solo. Os resultados indicam que a metodologia proposta é válida para os solos desse estudo. O sistema de manejo do solo indicou diferença significativa em D e S. Foram melhores no tratamento T2 comparado com T1 e T3 (D apresentou variação entre 0,00033 e 0,0321 cm2 s-1). Isso indica que o solo na condição T2 conduz água mais rápida na condição não saturada, que o solo manejado com pastejo. Assim, a metodologia proposta permite determinar D em uma amostra e com baixo custo, além de verificar a influência dessas propriedades em condições produtivas.Facultad de Ciencias Agrarias y Forestale