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We identified different lipemic and metabolic responses after the ingestion of a standardized meal by healthy adults and related them to atherosclerotic markers. Samples from 60 normolipidemic adults were collected before and after a liquid meal (40 g fat/m² body surface) at 0, 2, 4, 6, and 8 h for measurements of lipids, free fatty acids (FFA), insulin, cholesteryl ester transfer protein (CETP), autoantibodies to epitopes of oxidized LDL (oxLDL Ab), lipolytic activities, and apolipoprotein E polymorphism. Mean carotid intima-media thickness (cIMT) was determined by Doppler ultrasound. The volunteers were classified into early (N = 39) and late (N = 31) triacylglycerol (TAG) responders to the test meal. Late responders showed lower HDL cholesterol concentration at fasting and in the TAG peak, lower insulin and higher FFA concentrations compared to early responders. Multivariate regression analyses showed that mean cIMT was associated with gender (male) and age in early responders and by cholesterol levels at the 6th hour in late responders. oxLDL Ab were explained by lipoprotein lipase and negatively by hepatic lipase and oxLDL Ab (fasting period) by CETP (negative) and FFA (positive). This study is the first to identify a postalimentary insulin resistance state, combined with a reduced CETP response exclusively among late responders, and the identification of the regulators of postalimentary atherogenicity. Further research is required to determine the metabolic mechanisms described in the different postalimentary phenotypes observed in this study, as well as in different pathological states, as currently investigated in our laboratory.

Abnormal surface expression of HLA-DR by leukocytes is associated with a poor prognosis in critical care patients. Critical care patients often receive total parenteral nutrition with lipid emulsion (LE). In this study we evaluated the influence of fish oil LE (FO) on human monocyte/macrophage (Mφ) expression of surface HLA-DR under distinct activation states. Mononuclear leukocytes from the peripheral blood of healthy volunteers (n=18) were cultured for 24 hours without LE (control) or with 3 different concentrations (0.1, 0.25, and 0.5%) of the follow LE: a) pure FO b) FO in association (1:1-v/v) with LE composed of 50% medium-chain trygliceride and 50% soybean oil (MCTSO), and c) pure MCTSO. The leukocytes were also submitted to different cell activation states, as determinate by addition time: no INF-γ addition, 18 hours before, or at the time of LE addition. HLA-DR expression on Mφ surface was evaluated by flow cytometry using specific monoclonal antibodies. In relation to controls (for 0.1%, 0.25%, and 0.5%: 100) FO decreased the expression of HLA-DR when added alone [in simultaneously-activated Mφ, for 0.1%: 70 (59 ± 73); for 0.25%: 51 (48 ± 56); and for 0.5%: 52.5 (50 ± 58)] or in association with MCTSO [in simultaneously-activated Mφ, for 0.1%: 50.5 (47 ± 61); for 25%: 49 (45 ± 52); and for 0.5%: 51 (44 ± 54) and in previously-activated Mf, for 1.0%: 63 (44 ± 88); for 0.25%: 70 (41 ± 88); and for 0.5%: 59.5 (39 ± 79)] in culture medium (Friedman p < 0.05). In relation to controls (for 0.1%, 0.25%, and 0.5%: 100), FO did not influence the expression of these molecules on non-activated Mφ [for 0.1%: 87.5 (75±93); for 0.25%: 111 (98 ± 118); and for 0.5%: 101.5 (84 ± 113)]. Results show that parenteral FO modulates the expression of HLA-DR on human Mφ surface accordingly to leukocyte activation state. Further clinical studies evaluating the ideal moment of fish oil LE infusion to modulate leukocyte functions may contribute to a better understanding of its immune modulatory properties.La expresión anómala del HLA-DR sobre la superficie de los leucocitos se asocia con un pronóstico sombrío en pacientes críticos. A menudo, estos pacientes reciben nutrición parenteral total con una emulsión lipídica (EL). En este estudio, evaluamos la influencia de una EL de aceite de pescado (AP) en la expresión del HLA-DR en la superficie de monocitos/macrófagos humanos (MΦ) bajo diferentes estados de activación. Se cultivaron leucocitos mononucleares de sangre periférica de voluntarios sanos (n = 18) durante 24 horas sin la EL (control) o con 3 concentraciones distintas (0,1, 0,25 y 0,5%) de la siguiente EL: a) AP puro b) AP en asociación con EL (1:1 v/v) compuesta de 50% de triglicéridos de cadena media y 50% de aceite de soja (TCMAS), y c) TCMAS puro. También sometimos a los leucocitos a diferentes estados de activación celular, determinado por INF-γ tiempo de adición: no INF-γ adición 18 horas antes, o en el momento de añadir la EL. La expresión del HLA-DR sobre la superficie de los MΦ se evaluó mediante citometría de flujo utilizando anticuerpos monoclonales específicos. En relación con los controles (para 0,1%, 0,25% y 0,5%: 100) el AP disminuyó la expresión de HLA-DR cuando se añadía solo [en MΦ activados simultáneamente, para 0,1%: 70 (59 ± 73); para 0,25%: 51 (48 ± 56); y para 0,5%: 52.5 (50 ± 58)] o en asociación con TCMAS [en MΦ activados simultáneamente para 0,1%: 50,5 (47 ± 61); para 0,25%:(45 ± 52); y para 0,5%: 51 (44 ± 54) y en MΦ previamente activados para 0,1%: 63 (44 ± 88); para 0,25%: 70 (41 ± 88); y para 0,5%: 59,5 (39 ± 79)] en el medio de cultivo (Friedman p < 0,05). Con respecto a los controles, (para 0,1%, 0,25% y 0,5%: 100), el AP no influyó en la expresión de estas moléculas en MΦ no activados [para 0,1%: 87,5 (75 ± 93); para 0,25%: 111 (98 ± 118); y para 0,5%: 101,5 (84 ± 113)]. Los resultados muestran que el AP parenteral modula la expresión del HLA-DR sobre la superficie de los MΦ humanos en función del estado de activación leucocitaria. Estudios clínicos adicionales que evalúen el momento idóneo de añadir la infusión de EL con aceite de pescado para modular las funciones leucocitarias podrían contribuir a entender mejor sus propiedades inmunomoduladoras.

Abnormal surface expression of HLA-DR by leukocytes is associated with a poor prognosis in critical care patients. Critical care patients often receive total parenteral nutrition with lipid emulsion (LE). In this study we evaluated the influence of fish oil LE (FO) on human monocyte/macrophage (Mphi) expression of surface HLA-DR under distinct activation states. Mononuclear leukocytes from the peripheral blood of healthy volunteers (n = 18) were cultured for 24 hours without LE (control) or with 3 different concentrations (0.1, 0.25, and 0.5%) of the follow LE: a) pure FO b) FO in association (1:1 - v/v) with LE composed of 50% mediumchain trygliceride and 50% soybean oil (MCTSO), and c) pure MCTSO. The leukocytes were also submitted to different cell activation states, as determinate by INF-γ addition time: no INF-γ addition, 18 hours before, or at the time of LE addition. HLA-DR expression on Mphi surface was evaluated by flow cytometry using specific monoclonal antibodies. In relation to controls (for 0.1%, 0.25%, and 0.5%: 100) FO decreased the expression of HLA-DR when added alone [in simultaneously-activated Mphi, for 0.1%: 70 (59 ± 73); for 0.25%: 51 (48 ± 56); and for 0.5%: 52.5 (50 ± 58)] or in association with MCTSO [in simultaneously-activated Mphi, for 0.1%: 50.5 (47 ± 61); for 25%: 49 (45 ± 52); and for 0.5%: 51 (44 ± 54) and in previously-activated Mphi, for 1.0%: 63 (44 ± 88); for 0.25%: 70 (41 ± 88); and for 0.5%: 59.5 (39 ± 79)] in culture medium (Friedman p < 0.05). In relation to controls (for 0.1%, 0.25%, and 0.5%: 100), FO did not influence the expression of these molecules on non-activated Mphi [for 0.1%: 87.5 (75 ± 93); for 0.25%: 111 (98 ± 118); and for 0.5%: 101.5 (84 ± 113)]. Results show that parenteral FO modulates the expression of HLA-DR on human Mphi surface accordingly to leukocyte activation state. Further clinical studies evaluating the ideal moment of fish oil LE infusion to modulate leukocyte functions may contribute to a better understanding of its immune modulatory properties.La expresión anormal del HLA-DR en la superficie de los leucocitos se asocia con un pronóstico peor en los enfermos críticos. Estos enfermos a menudo reciben nutrición parenteral total con una emulsión lipídica (EL.) En este estudio evaluamos la influencia de la EL de aceite de pescado (AP) sobre la expresión del HLA-DR de superficie por los monocitos /macrófagos humanos (Mfi) en distintos estados de activación. Se cultivaron leucocitos mononucleares de sangre periférica de voluntarios sanos (n = 18) durante 24 horas sin EL (control) o con tres concentraciones diferentes (0,1, 0,25 y 0,5%) de la siguiente EL: a) AP puro b) AP en asociación (1:1 en v/v) con la EL compuesta de un 50% de triglicéridos de cadena media y 50% de aceite de soja (TCMAS), y c) TCMAS puro. Se sometió a los leucocitos a tres estados de activación diferentes, como venía determinado por el tiempo de adición de INF-γ: sin añadir INF-γ, 18 horas antes o en el momento de añadir la EL. La expresión de HLA-DR en la superficie de los Mfi se evaluó mediante citometría de flujo empleando anticuerpos monoclonales específicos. En relación con los controles (para 0,1%, 0,25% y 0,5%: 100) el AP disminuyó la expresión de HLA-DR cuando se añadía solo {en Mfi activados de forma simultánea, para 0,1%: 70 (59 ± 73); para 0,25%: 51 (48 ± 56) y para 0,5%: 52,5 (50 ± 58)} o en asociación con TCMAS [en Mfi activados de forma simultánea, para 0,1%: 50,5 (47 ± 61); para 25%: 49 (45 ± 52); y para 0,5%: 51 (44 ± 54) y en Mfiactivados previamente, para 1,0%: 63 (44 ± 88); para 0,25%: 70 (41 ± 88); y para 0,5%: 59,5 (39 ± 79)] en medio de cultivo (Friedman p < 0,05.) En relación con los controles (para 0,1%, 0,25% y 0,5%: 100), el AP no influyó en la expresión de estas moléculas en los Mfino activados [para 0,1%: 87,5 (75 ± Ó93); para 0,25%: 111 (98 ± 118); y para 0,5%: 101,5 (84 ± 113)}. Los resultados muestran que el AP parenteral modula la expresión del HLA-DR sobre la superficie de los Mfihumanos en función del estado de activación de los leucocitos. Estudios clínicos adicionales que evalúen el momento ideal de la infusión de la EL con aceite de pescado para modular las funciones leucocitarias podrían contribuir a un mejor conocimiento de sus propiedades inmunomoduladoras.

Background & aim: To compare the effect of fish oilbased (FO) lipid emulsions (LE) for parenteral administration with standard LE and a new FO containing LE composed of four different oils on the antigen presentation and inflammatory variables. Methods: Phytohemagglutinin (PHA) activated human mononuclear leukocytes were cultured with different LE - Control: without LE; SO: soybean oil; SO/FO: soybean and FO (4:1); MCT/SO: medium chain triglycerides and SO (1:1); MCT/SO/FO: MCT/SO and FO (4:1) and SMOF: a new LE containing FO. Cytokine production was evaluated by ELISA, the expression of antigen-presenting and co-stimulatory surface molecules were analyzed by flow cytometry and lymphocyte proliferation was assessed by H³-Thymidine incorporation, after tetanus toxoid-induced activation. Results: All LE decreased the HLA-DR and increased CD28 and CD152 expression on monocytes/macrophages and lymphocytes surface (p < 0.05). SO/FO and MCT/SO/FO decreased lymphocyte proliferation (pAntecedentes & objetivo: Comparar el efecto de las emulsiones lipídicas (EL) basadas en aceite de pescado (AP) para la administración parenteral con las EL estándar y una nueva EL que contiene AP compuesta por cuatro aceites distintos sobre la presentación antigénica y las variables inflamatorias. Métodos: se cultivaron leucocitos mononucleares activados con fitohemaglutinina (PHA) con diferentes EL - Control: sin EL; AS: aceite de soja; AS/AP: soja y AP (4:1); TCM/AS: triglicéridos de cadena media y AS (1:1); TCM/AS/AP: TCM/AS y AP (4:1) y SMOF: una nueva EL que contiene AP. Se evaluó la producción de citocinas mediante ELISA, se analizó la expresión de moléculas de superficie de presentación de antígeno y co-estimuladoras mediante citometría de flujo y se evaluó la proliferación linfocitaria mediante la incorporación de timidina-H³ tras la activación inducida por el toxoide tetánico. Resultados: Todas las EL disminuyeron la expresión de HLA-DR y aumentaron la expresión de CD28 y CD152 sobre superficie de monocitos/macrófagos y linfocitos (p < 0,05). La AS/AP y la TCM/AS/AP disminuyeron la proliferación linfocitaria (p < 0,05). Todas las EL disminuyeron la producción de IL-2, pero su efecto se incrementó con las emulsiones TCM/AS/AP y SMOF (p < 0,05). La TCM/AS/AP disminuyó la IL-6 y aumentó la IL-10, mientras que el AS tuvo el efecto opuesto (p < 0,05). Conclusión: La EL AP inhibió la proliferación linfocitaria y tuvo un efecto antiinflamatorio. Estos efectos parecen estar potenciados cuando el AP se mezcla con TCM/AS. La SMOF tuvo un efecto neutro sobre la proliferación linfocitaria y la producción de IL-6 e IL-10.

In this work, we investigated the impact of testosterone deficiency and cholesteryl ester transfer protein (CETP) expression on lipoprotein metabolism and diet-induced atherosclerosis. CETP transgenic mice and nontransgenic (nTg) littermates were studied 4 weeks after bilateral orchidectomy or sham operation. Castrated mice had an increase in the LDL fraction (+36% for CETP and +79% for nTg mice), whereas the HDL fraction was reduced (−30% for CETP and −11% for nTg mice). Castrated mice presented 1.7-fold higher titers of anti-oxidized LDL (Ox-LDL) antibodies than sham-operated controls. Plasma levels of CETP, lipoprotein lipase, and hepatic lipase were not changed by castration. Kinetic studies showed no differences in VLDL secretion rate, VLDL-LDL conversion rate, or number of LDL and HDL receptors. Competition experiments showed lower affinity of LDL from castrated mice for tissue receptors. Diet-induced atherosclerosis studies showed that testosterone deficiency increased by 100%, and CETP expression reduced by 44%, the size of aortic lesion area in castrated mice. In summary, testosterone deficiency increased plasma levels of apolipoprotein B-containing lipoproteins (apoB-LPs) and anti-OxLDL antibodies, decreased LDL receptor affinity, and doubled the size of diet-induced atherosclerotic lesions. The expression of CETP led to a milder increase of apoB-LPs and reduced atherosclerotic lesion size in testosterone-deficient mice.

The introduction of highly active antiretroviral therapy (HAART) for patients infected with HIV has significantly prolonged the life expectancy and to some extent has restored a functional immune response. However, the premature introduction of HAART has led to a significant and alarming increase in cardiovascular complications, including myocardial infarction and the appearance of abnormal distribution of body fat seen as lipodystrophy. One key element in the development of ischemic coronary artery disease is the presence of circulating and tissue-fixed modified low density lipoprotein (mLDL) that contributes to the initiation and progression of arterial lesions and to the formation of foam cells. Even though not completely elucidated, the most likely mechanism involves mLDL in the inflammatory response and the induction of a specific immune response against mLDL. Circulating antibodies against mLDL can serve as an indirect marker of the presence of circulating and vessel-fixed mLDL. In the present study, we measured antibodies to mLDL and correlated them with immune status (i.e., number of CD4+ T cells) in 59 HIV patients and with the clinical manifestation of lipodystrophy in 10 patients. We observed a significant reduction in anti-mLDL antibody levels related both to lipodystrophy and to an immunocompromised state in HIV patients. We speculate that these antibodies may explain in part the rapid development of ischemic coronary artery disease in some patients.

The uptake of lipids and lipoprotein particles by macrophages undergoes phagocytic activation and the formation of foam cells are key events in atherosclerosis. In this study we determined how intact high density lipoproteins (HDL) and apolipoproteins-HDL (removal of the lipid component from HDL, i.e., apoHDL) influence the phagocytosis of zymosan by mouse peritoneal macrophages. Zymosan particles preincubated together with lipoproteins or alone (control) were incubated with the macrophages. Phagocytosis activity was reported as the percent of macrophages that internalized three or more zymosan particles. HDL co-incubated with zymosan did not influence the over-all uptake of zymosan particles compared to apoHDL, which greatly enhanced the ability of the particle to be phagocytized (P

The serologic assay is an important tool in the diagnosis of leishmaniasis. One of the most commonly used tests is enzyme-linked immunosorbent assay (ELISA). Since total Leishmania promastigotes are used as antigen in the routine assay, false-positive reactions are frequent due to cross-reaction with sera from other diseases, mainly Chagas' disease. Therefore, an antigen that determines less cross-reactivity has been pursued for the serodiagnosis of leishmaniasis. In the present study we analyzed the use of recombinant Leishmania infantum heat shock protein (Hsp) 83 in ELISA for the serodiagnosis of cutaneous (N = 12) and mucocutaneous leishmaniasis (N = 14) and we observed the presence of anti-L. infantum Hsp 83 antibodies in all samples as well as anti-Leishmania total antigen antibodies. When cross-reactivity was tested, chronic Chagas' disease patients (N = 10) did not show any reactivity. Therefore, we consider this L. infantum Hsp 83 to be a good antigen for routine use for serodiagnosis of tegumentary leishmaniasis.

In order to study the role of natural killer (NK) cells during the early period of Leishmania infection, BALB/c mice were selectively and permanently depleted of NK cells by injection with 90Sr and subsequently infected with Leishmania (Leishmania) amazonensis (HSJD-1 strain). 90Sr is known to selectively deplete NK cells, leaving an intact T- and B-cell compartment and preserving the ability to produce both interferon alpha and IL-2. This method of depletion has advantages when compared with depletion using anti-NK cell monoclonal antibodies because the effect is permanent and neither activates complement nor provokes massive cell death. In the present study, after one month of treatment with 90Sr, the depletion of NK cells was shown by a more than ten-fold reduction in the cytotoxic activity of these cells: 2 x 106 spleen cells from NK-depleted animals were required to reach the same specific lysis of target cells effected by 0.15 x 106 spleen cells from normal control animals. The histopathology of the skin lesion at 7 days after Leishmania infection showed more parasites in the NK cell-depleted group. This observation further strengthens a direct role of NK cells during the early period of Leishmania infection.

:Objective and Design: Oxidized low-density lipoproteins (oxLDL) and protein fractions obtained by size exclusion chromatography of oxLDL were tested for vascular permeability effects on topical application to the hamster cheek pouch.Materials: The hamster cheek pouch was prepared for intravital microscopy observations of macromolecular leakage at post capillary venules (=leaks) with FITC-dextran as tracer.Treatment: OxLDL (0.1 mg/ml), PAF (platelet activation factor, 50–100 nM) and protein fractions of oxLDL (10 μg/ml) were applied topically to hamster cheek pouches.Results: Application of oxLDL and PAF resulted in reversible increases in the number of leaks. The PAF-antagonist WEB 2170, L-NAME and a β2-adrenoceptor agonist inhibited (P<0.01) almost completely the macromolecular leakage induced with oxLDL or PAF. Protein fractions were found to be more effective than unfractionated oxLDL in inducing plasma leakage as calculated on mg/ml-basis.Conclusion: Hamster oxLDL is a potent inducer of macromolecular leakage increase in the hamster cheek pouch microcirculation. The principal effect is mediated by PAF-like structures produced by the oxidation of the LDL-particle but oxLDL also contains low molecular weight proteins that could contribute to the overall vascular permeability increasing effect of ox LDL. [ABSTRACT FROM AUTHOR]

U937 is a monocytic cell-line originally derived from a histiocytic lymphoma. In serum-free medium the growth of U937 cells was stimulated by addition of low density lipoprotein (LDL). Methylation of LDL impaired its ability to be taken up in U937 cells as well as the capacity to stimulate the growth of these cells. Pretreatment of U937 cells with a monoclonal antibody against the LDL receptor was also found to completely block the growth-promoting effect of LDL. Exposure of U937 cells to liposomes with a lipid composition similar to that of LDL did not stimulate the growth rate. These findings demonstrate that growth of U937 cells under serum-free conditions is related to the amount of LDL ingested by the cells and that this uptake is mediated by binding of LDL to the LDL receptor. To determine if LDL-induced growth of U937 cells can be used to identify LDL with decreased binding to the LDL receptor, U937 cells were incubated with LDL isolated from a patient with familial defective apolipoprotein B-100 and from subjects with various lipoprotein phenotypes. LDL containing defective apolipoprotein B-100 was found to be less than half as effective as LDL from normolipidemic controls in stimulating growth of U937 cells. LDL isolated from patients with hyperlipoproteinemia type IIa and IV did not differ from normal LDL in their ability to promote growth of U937 cells. The present results suggest that LDL-induced growth of U937 cells may be used as an assay to identify defective receptor binding of LDL.

In this study, we aimed to examine the impact of high endurance training on vascular health parameters and immune-endocrine responses against modified low-density lipoprotein (LDL) particles. This observational, cross-sectional study included high endurance-trained and healthy non-trained subjects. Vascular ultrasound was used to assess vascular health parameters based on carotid intima-media thickness and endothelial function (flow-mediated dilation). Enzyme-linked immunosorbent assays were used to measure interleukin (IL)-8 and IL-10, autoantibody isotypes anti-oxidized LDL (oxLDL) and anti-apolipoprotein B (ApoB-D) peptide. Plasma levels of the corticosterone and 17 α-hydroxyprogesterone hormones were analyzed by mass spectrometry. This study enrolled 96 subjects, of whom 44 were high endurance trained and 52 were healthy non-trained individuals. Smaller carotid intima-media thickness values were observed in the high-endurance trained than in the healthy non-trained males, while no differences were observed between female groups. Flow-mediated dilation measurements did not differ by training or sex. The humoral immune responses to IgG anti-oxLDL and IgM anti-ApoB-D autoantibodies showed an isotype imbalance between the high-endurance trained and the non-trained groups. Immunoendocrine parameters showed inverse correlations between 17 α-hydroxyprogesterone concentrations and carotid intima-media thickness measurements. Direct correlations were found between IL-10 concentrations and flow-mediated dilation measurements. Chronic high-endurance exercise modulates immune-endocrine and vascular health parameters, in a sex-dependent manner., Competing Interests: The authors declare no conflict of interest., (Thieme. All rights reserved.)

Background: There are limited data about the influence of stent composition on immune responses after percutaneous coronary intervention (PCI)., Objective: The aim was to compare the effects of PCI with conventional cobalt-chromium bare metal stent (BMS) and drug-eluting stent (DES) implantation on the modulation of humoral and cellular immune responses., Methods: A randomised, single-centre, open pilot study involving patients with stable coronary artery disease eligible for PCI was performed. Blood samples were collected from the peripheral artery (PA) and the coronary sinus (CS) at baseline and 40 weeks following PCI. IgM and IgG autoantibodies (Abs), anti-oxLDL and anti-ApoB-D, as well as cytokine levels were evaluated by enzyme-linked immunosorbent assay., Results: A total of 30 patients of 60 years mean age were included, 68% of whom were men. At the nine-month follow-up, a modulation in the levels of cytokines and autoantibodies was observed in both stent type groups. However, no difference was observed in the modulation of these markers between stents., Conclusion: The stent type promotes modulations in cellular and humoral immune responses in the long-term, with differences in the magnitude of effects in specific immune responses.

Singlet molecular oxygen ( 1 O 2 ) has been reported in wide arrays of applications ranging from optoelectronic to photooxygenation reactions and therapy in biomedical proposals. It is also considered a major determinant of photodynamic therapy (PDT) efficacy. Since the direct excitation from the triplet ground state ( 3 O 2 ) of oxygen to the singlet excited state 1 O 2 is spin forbidden; therefore, a rational design and development of heterogeneous sensitizers is remarkably important for the efficient production of 1 O 2 . For this purpose, quantum dots (QDs) have emerged as versatile candidates either by acting individually as sensitizers for 1 O 2 generation or by working in conjunction with other inorganic materials or organic sensitizers by providing them a vast platform. Thus, conjoining the photophysical properties of QDs with other materials, e.g., coupling/combining with other inorganic materials, doping with the transition metal ions or lanthanide ions, and conjugation with a molecular sensitizer provide the opportunity to achieve high-efficiency quantum yields of 1 O 2 which is not possible with either component separately. Hence, the current review has been focused on the recent advances made in the semiconductor QDs, perovskite QDs, and transition metal dichalcogenide QD-sensitized 1 O 2 generation in the context of ongoing and previously published research work (over the past eight years, from 2015 to 2023)., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)

Background: Staphylococcus aureus is one of the most frequently major mastitis pathogens that cause clinical and subclinical mastitis worldwide. Current antimicrobial treatments are usually ineffective, and the commercially available vaccines lack proven effectiveness. The immunological response elicited by the recombinant S. aureus -cure-associated proteins phosphoglycerate kinase (PGK), enolase (ENO), and elongation factor-G (EF-G) in combination with the granulocyte-macrophage colony-stimulating factor (GM-CSF) DNA vaccination was studied in this work., Methods: Here, twenty-three C57BL/6 mice were divided into four groups and vaccinated with: G1: none (control); G2: GM-CSF DNA plasmid DNA vaccine; G3: the combination of EF-G+ENO+PGK; and G4: the combinations of EF-G+ENO+PGK proteins plus GM-CSF plasmid DNA vaccine. After 44 days, spleen cells were collected for immunophenotyping and lymphocyte proliferation evaluation by flow cytometry upon S. aureus stimulus., Results: Immunization with the three S. aureus recombinant proteins alone resulted in a higher percentage of IL-17A + cells among CD8 + T central memory cells, as well as the highest intensity of IL-17A production by overall lymphocytes indicating that the contribution of the combined lymphocyte populations is crucial to sustaining a type 3 cell immunity environment., Conclusion: The immunization with three S. aureus- cure-associated recombinant proteins triggered type 3 immunity, which is a highly interesting path to pursue an effective bovine S. aureus mastitis vaccine.

Staphylococcus aureus mastitis constitutes a serious threat to dairy cows. The reasons why available vaccines are not fully effective remain poorly understood; thus, in the present study, we investigated CD4 + and CD8 + T lymphocyte proliferation in dairy cows vaccinated with a polyvalent mastitis vaccine that had distinct precedent Staphylococcus aureus mastitis. We studied 17 S. aureus-infected dairy cows (11 vaccinated and six unvaccinated) and eight vaccinated healthy dairy cows with no previous S. aureus mastitis infections. Flow cytometry was used to assess lymphocyte proliferation using an anti-Ki67 antibody, and monoclonal antibodies were used to identify T cell subsets. S. aureus-infected cows exhibited reduced overall lymphocyte proliferation, including CD4 + T lymphocyte proliferation, and memory lymphocyte proliferation in response to S. aureus isolate stimulus. Immunization did not influence the expansion of blood lymphocyte populations. Furthermore, CD8 + T cells, memory CD8 + T lymphocytes, and effector memory CD8 + T lymphocytes displayed reduced proliferation 21 days after the third vaccine dose compared with before vaccination at time zero. The present data demonstrates an overall negative regulation of the T-cell response suggesting its detrimental impact leading to the persistence of S. aureus intramammary infections. Furthermore, the lack of vaccination effect on T-cell mediated immunity (e.g., proliferation) may be related to poor vaccine efficacy., Competing Interests: Declaration of interest None., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Group communication enables Internet of Things (IoT) devices to communicate in an efficient and fast manner. In most instances, a group message needs to be encrypted using a cryptographic key that only devices in the group know. In this paper, we address the problem of establishing such a key using a lattice-based one-way function, which can easily be inverted using a suitably designed lattice trapdoor. Using the notion of a bad/good basis, we present a new method of coupling multiple private keys into a single public key, which is then used for encrypting a group message. The protocol has the apparent advantage of having a conjectured resistance against potential quantum-computer-based attacks. All functions-key establishment, session key update, node addition, encryption, and decryption-are effected in constant time, using simple linear-algebra operations, making the protocol suitable for resource-constrained IoT networks. We show how a cryptographic session group key can be constructed on the fly by a user with legitimate credentials, making node-capture-type attacks impractical. The protocol also incorporates a mechanism for node addition and session-key generation in a forward- and backward-secrecy-preserving manner.

Background and Aims: Familial hypercholesterolemia (FH) is characterized by lifelong exposure to high LDL-c concentrations and premature atherosclerotic cardiovascular disease; nevertheless, disease severity can be heterogeneous.We aimed at evaluating if the immune-inflammatory system could modulate atherosclerosis burden in FH., Methods: From a cohort of subjects with confirmed FH (Dutch Lipid Clinic Network and genotype), 92 patients receiving high-intensity lipid-lowering therapy (statin ± ezetimibe) were included. The extension and severity of coronary atherosclerosis was assessed by standardized reporting systems (CAD-RADS) for coronary computed tomography angiography (CCTA) and coronary artery calcium (CAC) scores. Lipids, apolipoproteins, anti -oxLDL and anti-apolipoprotein B-D peptide ( anti -ApoB-D) autoantibodies (IgM and IgG), lymphocytes subtypes, platelet, monocyte and endothelial microparticles (MP), IgM levels (circulating or produced by B1 cells) and cytokines in the supernatant of cultured cells were determined. Multiple linear regression models evaluated associations of these biomarkers with CAC and CAD-RADS scores., Results: In univariate analysis CAC correlated with age, systolic blood pressure, TCD4+ cells, and titers of IgM anti -ApoB-D. In multiple linear regression [ANOVA F = 2.976; p  = 0.024; R 2  = 0.082), CD4+T lymphocytes (B = 35.289; beta = 0.277; p  = 0.010; 95%CI for B 8.727 to 61.851), was independently associated with CAC. CAD-RADS correlated with age, systolic blood pressure, titers of IgM anti -ApoB-D, and endothelial MP in univariate analysis. In multiple linear regression, [ANOVA F = 2.790; p  = 0.032; R 2  = 0.119), only age (B = 0.027; beta = 0.234; p  = 0.049; 95% CI for B 0.000 to 0.053) was independent predictor., Conclusions: In subjects with FH, under high-intensity lipid-lowering therapy, age and CD4+T cells were associated to atherosclerosis burden., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors.)

The development of QDs-based fluorescent bionanoprobe for cellular imaging fundamentally relies upon the precise knowledge of particle-cell interaction, optical properties of QDs inside and outside of the cell, movement of a particle in and out of the cell, and the fate of particle. We reported engineering and physicochemical characterization of water-dispersible Eu 3+ /Mn 2+ co-doped ZnSe@ZnS core/shell QDs and studied their potential as a bionanoprobe for biomedical applications, evaluating their biocompatibility, fluorescence behaviour by CytoViva dual mode fluorescence imaging, time-dependent uptake, endocytosis and exocytosis in RAW 264.7 macrophages. The oxidation state and local atomic structure of the Eu dopant studied by X-ray absorption fine structure (XAFS) analysis manifested that the Eu 3+ ions occupied sites in both ZnSe and ZnS lattices for the core/shell QDs. A novel approach was developed to relieve the excitation constraint of wide bandgap ZnSe by co-incorporation of Eu 3+ /Mn 2+ codopants, enabling the QDs to be excited at a wide UV-visible range. The QDs displayed tunable emission colors by a gradual increase in Eu 3+ concentration at a fixed amount of Mn 2+ , systematically enhancing the Mn 2+ emission intensity via energy transfer from the Eu 3+ to Mn 2+ ion. The ZnSe:Eu 3+ /Mn 2+ @ZnS QDs presented high cell viability above 85% and induced no cell activation. The detailed analyses of QDs-treated cells by dual mode fluorescence CytoViva microscopy confirmed the systematic color-tunable fluorescence and its intensity enhances as a function of incubation time. The QDs were internalized by the cells predominantly via macropinocytosis and other lipid raft-mediated endocytic pathways, retaining an efficient amount for 24 h. The unique color tunability and consistent high intensity emission make these QDs useful for developing a multiplex fluorescent bionanoprobe, activatable in wide-visible region.

Staphylococcus aureus mastitis remains a major challenge for dairy farming. Here, 24 mice were immunized and divided into four groups: G1: control; G2: Granulocyte Macrophage Colony-Stimulating Factor (GM-CSF) DNA vaccine; G3: F0F1 ATP synthase subunit α (SAS), succinyl-diaminopimelate (SDD), and cysteinyl-tRNA synthetase (CTS) recombinant proteins; and G4: SAS+SDD+CTS plus GM-CSF DNA vaccine. The lymphocyte subpopulations, and the intracellular interleukin-17A (IL-17A) and interferon-γ production in the draining lymph node cells were immunophenotyped by flow cytometry. The immunophenotyping and lymphocyte proliferation was determined in spleen cells cultured with and without S. aureus stimulus. Immunization with S. aureus recombinant proteins generated memory cells in draining lymph nodes. Immunization with the three recombinant proteins plus GM-CSF DNA led to an increase in the percentage of IL-17A + cells among overall CD44 + (memory), T CD4 + , CD4 + T CD44 + CD27 - , γδ TCR, γδ TCR + CD44 + CD27 + , and TCRVγ4 + cells. Vaccination with S. aureus recombinant proteins associated with GM-CSF DNA vaccine downregulated T H 2 immunity. Immunization with the three recombinant proteins plus the GM-CSF DNA led to a proliferation of overall memory T, CD4 + , and CD4 + TEM cells upon S. aureus stimulus. This approach fostered type 3 immunity, suggesting the development of a protective immune response against S. aureus .

The increasing proliferation of Internet-of-things (IoT) networks in a given space requires exploring various communication solutions (e.g., cooperative relaying, non-orthogonal multiple access, spectrum sharing) jointly to increase the performance of coexisting IoT systems. However, the design complexity of such a system increases, especially under the constraints of performance targets. In this respect, this paper studies multiple-access enabled relaying by a lower-priority secondary system, which cooperatively relays the incoming information to the primary users and simultaneously transmits its own data. We consider that the direct link between the primary transmitter-receiver pair uses orthogonal multiple access in the first phase. In the second phase, a secondary transmitter adopts a relaying strategy to support the direct link while it uses non-orthogonal multiple access (NOMA) to serve the secondary receiver. As a relaying scheme, we propose a piece-wise and forward (PF) relay protocol, which, depending on the absolute value of the received primary signal, acts similar to decode-and-forward (DF) and amplify-and-forward (AF) schemes in high and low signal-to-noise ratio (SNR), respectively. By doing so, PF achieves the best of these two relaying protocols using the adaptive threshold according to the transmitter-relay channel condition. Under PF-NOMA, first, we find the achievable rate region for primary and secondary receivers, and then we formulate an optimization problem to derive the optimal PF-NOMA time and power fraction that maximize the secondary rate subject to reliability constraints on both the primary and the secondary links. Our simulation results and analysis show that the PF-NOMA outperforms DF-NOMA and AF-NOMA-based relaying techniques in terms of achievable rate regions and rate-guaranteed relay locations.

Backgorund: Traditional and HIV-defined risk factors may be associated with an increase in cardiovascular events. Recent studies have suggested that the humoral immune response to modified LDL may be associated with the process of atherosclerosis., Objectives: To evaluate the presence of anti-oxLDL and apolipoprotein B-derived peptides in the blood, and their association with the endothelial function in HIV-infection., Methods: This study consecutively included subjects matched for age, gender, and demographic data in two groups: (1) HIV-infected and naïve for antiviral therapy and (2) uninfected individuals. Subclinical atherosclerosis was assessed by intimal-media thickness, using ultrasonography of the carotid arteries. Endothelial function was determined by flow-mediated dilatation (FMD) of the brachial artery by ultrasonography. Autoantibodies (IgM, IgG) anti-oxidized low-density lipoprotein (oxLDL), anti-apolipoprotein B-peptide fragments (ApoB-D and 0033G-Cys peptides), and cytokine levels were evaluated by ELISA., Results: This study's results showed no difference in subclinical atherosclerosis between groups; however, HIV-infected subjects showed a lower FMD, when compared to non-infected subjects. Therefore, HIV-infected subjects showed higher levels of inflammatory cytokines, titers of IgG anti-oxLDL, and IgG anti-ApoB-D. In contrast, titers of IgM anti-ApoB-D were lower in HIV-infected individuals and associated with reduced endothelial functions., Conclusions: This study's results show that HIV infection, in naïve subjects, is associated with endothelial dysfunction and a decline of natural antibodies to apo-B antigens.

The Routing Protocol for Low power and lossy networks (RPL) has been introduced as the de-facto routing protocol for the Industrial Internet of Things (IIoT). In heavy load scenarios, particular parent nodes are likely prone to congestion, which in turn degrades the network performance, in terms of packet delivery and delay. Moreover, there is no explicit strategy in RPL to prioritize the transmission of different traffic types in heterogeneous 6TiSCH networks, each according to its criticality. In this paper, we address the aforementioned issues by introducing a congestion control and service differentiation strategies to support heterogeneous 6TiSCH networks in IIoT applications. First, we introduce a congestion control mechanism to achieve load balancing under heavy traffic scenarios. The congestion is detected through monitoring and sharing the status of the queue backlog among neighbor nodes. We define a new routing metric that considers the queue occupancy when selecting the new parent node in congestion situations. In addition, we design a multi-queue model to provide prioritized data transmission for critical data over the non-critical ones. Each traffic type is placed in a separate queue and scheduled for transmission based on the assigned queue priority, where critical data are always transmitted first. The performance of the proposed work is evaluated through extensive simulations and compared with existing work to demonstrate its effectiveness. The results show that our proposal achieves improved packet delivery and low queue losses under heavy load scenarios, as well as improved delay performance of critical traffic.

Staphylococcus aureus is one of the pathogens most frequently isolated from cases of mastitis worldwide. To decrease the effect of S. aureus mastitis in dairy farming, alternative strategies for controlling mastitis are needed that depend on a better knowledge of cow-to-cow variations in S. aureus antibody production. The present study sought to explore the diversity of S. aureus antibodies produced by dairy cows with a distinct mastitis history and vaccinated with a polyvalent mastitis vaccine. We obtained protein extracts from S. aureus isolates derived from persistent subclinical mastitis. Proteins were fractionated using 2-dimensional gel electrophoresis and Western blotting. Then, Western blotting membranes were exposed to sera from 24 dairy cows that had been divided into the following groups: vaccinated dairy cows that were infected with S. aureus, further subdivided according to whether they (a) remained infected by S. aureus or (b) recovered from the intramammary infection; unvaccinated dairy cows infected with S. aureus; and vaccinated healthy dairy cows with no history of S. aureus mastitis. Proteins found to be reactive by Western blot were identified by mass spectrometry (MALDI/TOF-TOF). Our most important finding was that F 0 F 1 ATP synthase subunit α, succinyl-diaminopimelate desuccinylase, and cysteinyl-tRNA synthetase were potential candidate proteins for the prevention of S. aureus mastitis. This study strengthens the notion that variations among animals should not be ignored and shows that the heterogeneity of antibody production against anti-staphylococcal antigens in animals may enable the identification of new immunotherapy targets., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Industry is going through a transformation phase, enabling automation and data exchange in manufacturing technologies and processes, and this transformation is called Industry 4.0. Industrial Internet-of-Things (IIoT) applications require real-time processing, near-by storage, ultra-low latency, reliability and high data rate, all of which can be satisfied by fog computing architecture. With smart devices expected to grow exponentially, the need for an optimized fog computing architecture and protocols is crucial. Therein, efficient, intelligent and decentralized solutions are required to ensure real-time connectivity, reliability and green communication. In this paper, we provide a comprehensive review of methods and techniques in fog computing. Our focus is on fog infrastructure and protocols in the context of IIoT applications. This article has two main research areas: In the first half, we discuss the history of industrial revolution, application areas of IIoT followed by key enabling technologies that act as building blocks for industrial transformation. In the second half, we focus on fog computing, providing solutions to critical challenges and as an enabler for IIoT application domains. Finally, open research challenges are discussed to enlighten fog computing aspects in different fields and technologies.

With recent adoption of Wireless Sensor-Actuator Networks (WSANs) in industrial automation, wireless control systems have emerged as a frontier of industrial networks. Hence, it has been shown that existing standards and researches concentrate on the reliability and real-time performance of WSANs. The multipath retransmission scheme with multiple channels is a key approach to guarantee the deterministic wireless communication. However, the efficiency of resource scheduling is seldom considered in applications with diverse data sampling rates. In this paper, we propose an efficient resources scheduling algorithm for multipath retransmission in WSANs. The objective of our algorithm is to improve efficiency and schedulability for the use of slot and channel resources. In detail, the proposed algorithm uses the approaches of CCA (clear channel assessment)-Embedded slot and Multiple sinks with Rate Monotonic scheme (CEM-RM) to decrease the number of collisions. We have simulated and implemented our algorithm in hardware and verified its performance in a real industrial environment. The achieved results show that the proposed algorithm significantly improves the schedulability without trading off reliability and real-time performance.