RESUMO
BACKGROUND: Immunosuppressive therapy is the standard management of adults with aplastic anaemia. Antithymocyte globulin is used as first-line treatment of patients not eligible for bone marrow transplantation. This being a rare disease, available evidence in India is scarce. This study aimed to present experience in treating adult aplastic anaemia patients by immunosuppressive therapy using antithymocyte globulin-equine (Thymogam) in two tertiary care centres of northeast India. METHODS: This case series was conducted at the Health city hospital, Guwahati, and Excel Care Hospital, Guwahati from 2018 to 2020. Eighteen adult aplastic anaemia patients who were treated by immunosuppressive therapy with antithymocyte globulin-equine (Thymogam) and followed up for two years were included. Treatment response and relapse are described. RESULTS: All the 18 patients, (14 severe, four very severe) were uniformly treated with immunosuppressive therapy (Thymogam 40 mg/kg/d for four days with oral Cyclosporine from Day-1). Cyclosporin A was used as a concomitant drug in 94.44 % of the patients. At two years of follow up, 66.7 % showed a response and the mortality rate was 11.1 %. CONCLUSION: The results of this case series substantiate the effectiveness of immunosuppressive therapy with a low-cost preparation of horse antithymocyte globulin (Thymogam) along with cyclosporin A in the management of aplastic anaemia patients not suitable for bone marrow transplantation.
RESUMO
Amblyomin-X is a Kunitz-type FXa inhibitor identified through the transcriptome analysis of the salivary gland from Amblyomma sculptum tick. This protein consists of two domains of equivalent size, triggers apoptosis in different tumor cell lines, and promotes regression of tumor growth, and reduction of metastasis. To study the structural properties and functional roles of the N-terminal (N-ter) and C-terminal (C-ter) domains of Amblyomin-X, we synthesized them by solid-phase peptide synthesis, solved the X-Ray crystallographic structure of the N-ter domain, confirming its Kunitz-type signature, and studied their biological properties. We show here that the C-ter domain is responsible for the uptake of Amblyomin-X by tumor cells and highlight the ability of this domain to deliver intracellular cargo by the strong enhancement of the intracellular detection of molecules with low cellular-uptake efficiency (p15) after their coupling with the C-ter domain. In contrast, the N-ter Kunitz domain of Amblyomin-X is not capable of crossing through the cell membrane but is associated with tumor cell cytotoxicity when it is microinjected into the cells or fused to TAT cell-penetrating peptide. Additionally, we identify the minimum length C-terminal domain named F2C able to enter in the SK-MEL-28 cells and induces dynein chains gene expression modulation, a molecular motor that plays a role in the uptake and intracellular trafficking of Amblyomin-X.
RESUMO
We have investigated Amblyomin-X-treated horse melanomas to better understand its mode of action through transcriptome analysis and the in vivo model. Amblyomin-X is a Kunitz-type homologous protein that selectively leads to the death of tumor cells via ER stress and apoptosis, currently under investigation as a new drug candidate for cancer treatment. Melanomas are immunogenic tumors, and a better understanding of the immune responses is warranted. Equine melanomas are spontaneous and not so aggressive as human melanomas are, as this study shows that the in vivo treatment of encapsulated horse melanoma tumors led to a significant reduction in the tumor size or even the complete disappearance of the tumor mass through intratumoral injections of Amblyomin-X. Transcriptome analysis identified ER- and mitochondria-stress, modulation of the innate immune system, apoptosis, and possibly immunogenic cell death activation. Interactome analysis showed that Amblyomin-X potentially interacts with key elements found in transcriptomics. Taken together, Amblyomin-X modulated the tumor immune microenvironment in different ways, at least contributing to induce tumor cell death.
Assuntos
Antineoplásicos/uso terapêutico , Proteínas de Artrópodes/uso terapêutico , Doenças dos Cavalos/tratamento farmacológico , Melanoma/veterinária , Proteínas e Peptídeos Salivares/uso terapêutico , Animais , Morte Celular/efeitos dos fármacos , Descoberta de Drogas , Cavalos , Masculino , Melanoma/tratamento farmacológico , Microambiente Tumoral/efeitos dos fármacosRESUMO
In a tumor microenvironment, endothelial cell migration and angiogenesis allow cancer to spread to other organs causing metastasis. Indeed, a number of molecules that are involved in cytoskeleton re-organization and intracellular signaling have been investigated for their effects on tumor cell growth and metastasis. Alongside that, Amblyomin-X, a recombinant Kunitz-type protein, has been shown to reduce metastasis and tumor growth in in vivo experiments. In the present report, we provide a mechanistic insight to these antitumor effects, this is, Amblyomin-X modulates Rho-GTPases and uPAR signaling, and reduces the release of MMPs, leading to disruption of the actin cytoskeleton and decreased cell migration of tumor cell lines. Altogether, our data support a role for Amblyomin-X as a novel potential antitumor drug. ABBREVIATIONS: Amb-X: Amblyomin-X; ECGF: endotelial cell growth factor; ECM: extracellular matrix; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; HUVEC: human umbilical vein endothelial cell; LRP1: low-density lipoprotein receptor-related protein; MMP: matrix metalloproteinase; HPI-4: hedgehog pathway inhibitor 4; PAI-1: plasminogen activator inhibitor 1; PMA: phorbol 12-myristate-13-acetate; TFPI: tissue factor pathway inhibitor; uPA: urokinase plasminogen activator; uPAR: uPA receptor.
Assuntos
Aprotinina/farmacologia , Proteínas de Artrópodes/farmacologia , Movimento Celular/efeitos dos fármacos , Proteínas Recombinantes/farmacologia , Proteínas e Peptídeos Salivares/farmacologia , Adesão Celular/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/metabolismo , Humanos , Metaloproteinases da Matriz/metabolismo , Receptores de Ativador de Plasminogênio Tipo Uroquinase/metabolismo , Proteínas rho de Ligação ao GTP/metabolismoRESUMO
We have investigated Amblyomin-X-treated horse melanomas to better understand its mode of action through transcriptome analysis and the in vivo model. Amblyomin-X is a Kunitz-type homologous protein that selectively leads to the death of tumor cells via ER stress and apoptosis, currently under investigation as a new drug candidate for cancer treatment. Melanomas are immunogenic tumors, and a better understanding of the immune responses is warranted. Equine melanomas are spontaneous and not so aggressive as human melanomas are, as this study shows that the in vivo treatment of encapsulated horse melanoma tumors led to a significant reduction in the tumor size or even the complete disappearance of the tumor mass through intratumoral injections of Amblyomin-X. Transcriptome analysis identified ER- and mitochondria-stress, modulation of the innate immune system, apoptosis, and possibly immunogenic cell death activation. Interactome analysis showed that Amblyomin-X potentially interacts with key elements found in transcriptomics. Taken together, Amblyomin-X modulated the tumor immune microenvironment in different ways, at least contributing to induce tumor cell death.
RESUMO
Restoration, preservation and sustainability of agricultural resources including soil demands biologically and economically viable farming practices. For boosting physico-chemical properties of soil from 2012 to 2015, three legumes including cowpea, cluster bean and soybean were intercropped with sorghum in 40-60, 75-25 and 100-100 seed blending ratios, while pure stand of sorghum was kept for comparison. Correlation studies were also conducted to determine linear or inverse relationship between physiological and agronomic attributes with green forage yield of sorghum. Results revealed that intercropping systems were not effective in significantly improving the physical properties of soil, however soybean and sorghum mixed seeded crop in 100-100 seed ratio resulted in the highest total and available nitrogen especially during last two years of the study and it was followed by mixed seeded crop of cluster bean and sorghum (100-100 seed ratio), while pure stand of sorghum appeared to be the most exhaustive farming system. Correlation analysis depicted a linear relationship between all agronomic attributes (seedling count, plant population at harvest, plant height, stem diameter, number of leaves and leaf area per plant, fresh and dry weights per plant) and physiological growth parameters (leaf area index and leaf area duration, crop growth rate and net assimilation rate) with forage yield of sorghum. Thus, legumes inclusion was found to be effective in restoring soil fertility, while long term legumes inclusion by following rotation may be suggested.
A restauração, a preservação e a sustentabilidade dos recursos agrícolas, incluindo o solo, exigem práticas agrícolas biologicamente e economicamente viáveis. Para aumentar as propriedades físico-químicas do solo de 2012 a 2015, três leguminosas, incluindo feijão-fradinho, guar e soja, foram consorciadas com sorgo nas proporções de mistura de 40-60, 75-25 e 100-100, enquanto que o sorgo puro foi mantido por comparação. Estudos de correlação também foram conduzidos para determinar a relação linear ou inversa entre atributos fisiológicos e agronômicos com a produtividade de forragem verde do sorgo. Os resultados revelaram que os sistemas consorciados não foram eficazes em afetar significativamente as propriedades físicas do solo, no entanto, soja e sorgo misturados na proporção de sementes 100-100 resultou no maior nitrogênio total e nitrogênio disponível, especialmente durante os últimos dois anos do estudo e foi seguido do plantio de sementes mistas de guar e sorgo (100-100 rácio de semente), enquanto a monocultura de sorgo parecia ser o sistema de agricultura mais exaustivo. A análise de regressão mostrou uma correlação linear entre todos os atributos agronômicos (contagem de plântulas, população de plantas na colheita, altura de planta, diâmetro de caule, número de folhas e área foliar por planta, peso fresco e seco por planta) e parâmetros de crescimento fisiológico (índice de área foliar e duração da área foliar, taxa de crescimento da cultura e taxa de assimilação líquida) com o rendimento de forragem do sorgo. Assim, a inclusão de leguminosas foi eficaz no aumento da fertilidade do solo, enquanto a inclusão de leguminosas a longo prazo por rotação de diferentes leguminosas é sugerida.
Assuntos
Produção Agrícola , Qualidade do Solo , Sorghum , Matéria Orgânica , Nitrogênio , Glycine max , Agricultura , Cyamopsis , Vigna , FabaceaeRESUMO
In a tumor microenvironment, endothelial cell migration and angiogenesis allow cancer to spread to other organs causing metastasis. Indeed, a number of molecules that are involved in cytoskeleton re-organization and intracellular signaling have been investigated for their effects on tumor cell growth and metastasis. Alongside that, Amblyomin-X, a recombinant Kunitz-type protein, has been shown to reduce metastasis and tumor growth in in vivo experiments. In the present report, we provide a mechanistic insight to these antitumor effects, this is, Amblyomin-X modulates Rho-GTPases and uPAR signaling, and reduces the release of MMPs, leading to disruption of the actin cytoskeleton and decreased cell migration of tumor cell lines. Altogether, our data support a role for Amblyomin-X as a novel potential antitumor drug.
RESUMO
Background: Mutations in genes affecting interferon-γ (IFN-γ) immunity have contributed to understand the role of IFN-γ in protection against intracellular pathogens. However, inborn errors in STAT4, which controls interleukin-12 (IL-12) responses, have not yet been reported. Our objective was to determine the genetic defect in a family with a history of paracoccidioidomycosis. Methods: Genetic analysis was performed by whole-exome sequencing and Sanger sequencing. STAT4 phosphorylation (pSTAT4) and translocation to the nucleus, IFN-γ release by patient lymphocytes, and microbicidal activity of patient monocytes/macrophages were assessed. The effect on STAT4 function was evaluated by site-directed mutagenesis using a lymphoblastoid B cell line (B-LCL) and U3A cells. Results: A heterozygous missense mutation, c.1952 A>T (p.E651V) in STAT4 was identified in the index patient and her father. Patient's and father's lymphocytes showed reduced pSTAT4, nuclear translocation, and impaired IFN-γ production. Mutant B-LCL and U3A cells also displayed reduced pSTAT4. Patient's and father's peripheral blood mononuclear cells and macrophages demonstrated impaired fungicidal activity compared with those from healthy controls that improved in the presence of recombinant human IFN-γ, but not rhIL-12. Conclusion: Our data suggest autosomal dominant STAT4 deficiency as a novel inborn error of IL-12-dependent IFN-γ immunity associated with susceptibility to paracoccidioidomycosis.
Assuntos
Predisposição Genética para Doença , Interferon gama/deficiência , Subunidade p35 da Interleucina-12/metabolismo , Mutação de Sentido Incorreto , Paracoccidioidomicose/genética , Fator de Transcrição STAT4/genética , Adulto , Idoso , Linhagem Celular , Saúde da Família , Feminino , Genótipo , Heterozigoto , Humanos , Linfócitos/imunologia , Macrófagos/imunologia , Masculino , Análise de Sequência de DNARESUMO
Thrombin is a multifunctional enzyme with a key role in the coagulation cascade. Its functional modulation can culminate into normal blood coagulation or thrombosis. Thus, the identification of novel potent inhibitors of thrombin are of immense importance. Sculptin is the first specific thrombin inhibitor identified in the transcriptomics analysis of tick's salivary glands. It consists of 168 residues having four similar repeats and evolutionary diverged from hirudin. Sculptin is a competitive, specific and reversible inhibitor of thrombin with a Ki of 18.3 ± 1.9 pM (k on 4.04 ± 0.03 × 107 M-1 s-1 and k off 0.65 ± 0.04 × 10-3 s-1). It is slowly consumed by thrombin eventually losing its activity. Contrary, sculptin is hydrolyzed by factor Xa and each polypeptide fragment is able to inhibit thrombin independently. A single domain of sculptin alone retains ~45% of inhibitory activity, which could bind thrombin in a bivalent fashion. The formation of a small turn/helical-like structure by active site binding residues of sculptin might have made it a more potent thrombin inhibitor. In addition, sculptin prolongs global coagulation parameters. In conclusion, sculptin and its independent domain(s) have strong potential to become novel antithrombotic therapeutics.
Assuntos
Fibrinolíticos/química , Hirudinas/química , Fragmentos de Peptídeos/química , Peptídeos/química , Trombose/prevenção & controle , Animais , Ligação Competitiva , Coagulação Sanguínea/fisiologia , Domínio Catalítico , Cristalografia por Raios X , Fator Xa/química , Fator Xa/metabolismo , Fibrinolíticos/metabolismo , Expressão Gênica , Hirudinas/genética , Hirudinas/metabolismo , Humanos , Hidrólise , Ixodidae/química , Cinética , Modelos Moleculares , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/metabolismo , Peptídeos/genética , Peptídeos/metabolismo , Filogenia , Ligação Proteica , Conformação Proteica em alfa-Hélice , Conformação Proteica em Folha beta , Domínios e Motivos de Interação entre Proteínas , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Homologia Estrutural de Proteína , Trombose/sangue , Trombose/patologiaRESUMO
Heat stress during reproductive and grain filling phases adversely affects the growth of cereals through reduction in grains number and size. However, exogenous application of antioxidants, plant growth regulators and osmoprotectants may be helpful to minimize these heat induced yield losses in cereals. This two year study was conducted to evaluate the role of exogenous application of ascorbic acid (AsA), salicylic acid (SA) and hydrogen peroxide (H2 O2 ) applied through seed priming or foliar spray on biochemical, physiological, morphological and yield related traits, grain yield and quality of late spring sown hybrid maize. The experiment was conducted in the spring season of 2007 and 2008. We observed that application of AsA, SA and H2 O2 applied through seed priming or foliar spray improved the physiological, biochemical, morphological and yield related traits, grain yield and grain quality of late spring sown maize in both years. In both years, we observed higher superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activity in the plants where AsA, SA and H2 O2 were applied through seed priming or foliar spray than control. Membrane stability index (MSI), relative water contents (RWC), chlorophyll contents, grain yield and grain oil contents were also improved by exogenous application of AsA, SA and H2 O2 in both years. Seed priming of AsA, SA and H2 O2 was equally effective as the foliar application. In conclusion, seed priming with AsA, SA and H2 O2 may be opted to lessen the heat induced yield losses in late sown spring hybrid maize. Heat tolerance induced by ASA, SA and H2 O2 may be attributed to increase in antioxidant activities and MSI which maintained RWC and chlorophyll contents in maize resulting in better grain yield in heat stress conditions.(AU)
O estresse térmico durante as fases reprodutiva e de enchimento de grãos afeta negativamente o crescimento de cereais com redução do número e do tamanho do grão. A aplicação exógena de antioxidantes, reguladores de crescimento vegetal e osmoprotetores, pode ser útil para minimizar essas perdas. O objetivo foi avaliar o efeito da aplicação exógena de ácido ascórbico (ASA), ácido salicílico (SA) e peróxido de hidrogênio (H2 O2 ) aplicada através de sementes ou pulverização foliar na indução de tolerância ao estresse térmico em milho híbrido semeado no final da primavera. A aplicação de AsA, SA e H2 O2 através de sementes ou pulverização foliar melhora a fisiologia, bioquímica, morfologia e traços associados ao rendimento e qualidade de de grãos de milho no cultivo de Primavera tardia em ambos os anos. Foi observada maior superóxido dismutase (SOD), catalase (CAT) e atividade de peroxidase (POD) em plantas onde AsA, SA e H2 O2 foram aplicados via sementes ou pulverização foliar em relação ao controle. O índice de estabilidade de membrana (MSI), conteúdo relativo de água (RWC), conteúdo de clorofila, rendimento de grãos e teor de óleo de grãos também foram melhorados pela aplicação de AsA, exógena SA e H2 O2 em ambos os anos. A aplicação via sementes de AsA, SA e H2 O2 foi igualmente eficaz à aplicação foliar. A embebição de sementes com AsA, SA e H2 O2 pode ser utilizado para diminuir as perdas de rendimento induzida pelo calor em milho híbrido semeado no final da primavera. A indução de tolerância ao calor pela ASA, SA e H2 O2 pode ser atribuído ao aumento de atividades antioxidantes e a MSI que manteve o RWC e o conteúdo de clorofila resultando em melhor rendimento de grãos em condições de estresse de calor.(AU)