Antimicrobial evaluation of silver nanoparticles using extracts of Crescentia cujete L.

New natural reducing agents with a lower negative impact on the environment and with a high antimicrobial potential are required for the process of obtaining silver nanoparticles through the chemical reduction method. The use of plant extracts can be a fast track in the formation of nanoparticles. In this case, organic compounds such as terpenes, flavonoids, enzymes, proteins, and cofactors present in plants act as reducing agents for nanomaterials. This research evaluated the antimicrobial property of silver nanoparticles from extracts of Crescentia cujete L. The presence of quercetin (flavonoid) was determined by high-performance liquid chromatography (HPLC); the production of silver nanoparticles (AgNPs) was established by green synthesis; the size and morphology of the nanomaterials were evaluated by scanning electron microscope (SEM). The antimicrobial capacity was studied by two analysis methods: modified culture medium and surface seeding. The presence of quercetin (26.55 mg L-1) in the crude extract of Crescentia cujete L., identified by HPLC, was evidenced. Nanoparticle formation was spherical, with an average size of 250 ± 3 and 460 ± 6 nm. Microbiological cultures with treatment showed 94% microbial inhibition. It was concluded that the Crescentia cujete L., leaves shoed an acceptable concentration of quercetin to be used as a useful adjuvant to enhance the reduction of NPs synthesis. The nanoparticles produced by green synthesis proved to have a positive effect to combat pathogenic microorganisms.

ANÁLISIS DEL ESTADO ACTUAL DE LA INGENIERÍA CLÍNICA EN LAS INSTITUCIONES HOSPITALARIAS DE CALI / ANALYSIS OF THE CURRENT STATE OF CLINICAL ENGINEERING IN HOSPITALS IN CALI / A ANÁLISE DO ESTADO ATUAL DA ENGENHARIA CLÍNICA EM HOSPITAIS DE CALI

En este trabajo se presentan los resultados obtenidos tras analizar la información recolectada en once instituciones prestadoras de servicios de salud (IPS) en la ciudad de Cali y municipios aledaños, sobre tres elementos clave para la buena práctica de la Ingeniería Clínica: adquisición de tecnología, gestión de mantenimiento y formación del personal. Se realizó una comparación entre las prácticas actuales de IC en las IPS encuestadas y las prácticas propuestas en la literatura existente. Se propone además una serie de aspectos a tomar en cuenta con miras a mejorar el desempeño de los departamentos de IC tanto en la ciudad como en el país.

This paper summarizes the results obtained after analyzing the information gathered in eleven institutions providing health services (IPS, in Colombia) in the city of Cali and surrounding municipalities, three key elements for good clinical engineering practice are presented: Acquisition technology, management, maintenance and staff training. A comparison between current practices in the surveyed IC at IPS and practices proposed in the literature was conducted. It also proposes a number of aspects to consider in order to improve performance of both IC departments in the city and in the country.

Este artigo apresenta os resultados obtidos depois de analisar as informações coletadas em onze instituições prestadoras de serviços de saúde (IPS), na cidade de Cali e municípios vizinhos, sobre três elementos-chave para a boa prática de engenharia clínica: aquisição de tecnologia, gestão de manutenção e treinamento de pessoal. Uma comparação foi feita entre as práticas atuais nas IPS e práticas propostos na literatura atual. Ele também propõe uma série de aspectos a considerar, a fim de melhorar o desempenho dos departamentos IC tanto na cidade quanto no país.

The SH2-containing inositol-5'-phosphatase enhances LFA-1-mediated cell adhesion and defines two signaling pathways for LFA-1 activation.

The inside-out signaling involved in the activation of LFA-1-mediated cell adhesion is still poorly understood. Here we examined the role of the SH2-containing inositol phosphatase (SHIP), a major negative regulator of intracellular signaling, in this process. Wild-type SHIP and a phosphatase-deficient mutant SHIP were overexpressed in the murine myeloid cell line, DA-ER, and the effects on LFA-1-mediated cell adhesion to ICAM-1 (CD54) were tested. Overexpression of wild-type SHIP significantly enhanced cell adhesion to immobilized ICAM-1, and PMA, IL-3, or erythropoietin further augmented this adhesion. In contrast, phosphatase dead SHIP had no enhancing effects. Furthermore, PMA-induced activation of LFA-1 on DA-ER cells overexpressing wild-type SHIP was dependent on protein kinase C but independent of mitogen-activated protein kinase activation, whereas cytokine-induced activation was independent of protein kinase C and mitogen-activated protein kinase activation but required phosphatidylinositol-3 kinase activation. These results suggest that SHIP may regulate two distinct inside-out signaling pathways and that the phosphatase activity of SHIP is essential for both of them.

Dominant-negative effect of the lymphocyte function-associated antigen-1 beta (CD18) cytoplasmic domain on leukocyte adhesion to ICAM-1 and fibronectin.

The cytoplasmic domains of LFA-1 (CD11a/CD18) are thought to play an important role in the regulation of LFA-1 function. To further elucidate the role of the LFA-1 cytoplasmic domains, we transfected chimeric proteins consisting of the extracellular domain of CD4 fused with the transmembrane and cytoplasmic domains of LFA-1 into T and B cell lines, EL-4 and A20, respectively, and examined their effects on LFA-1-mediated cell adhesion. The CD4/18, but not CD4/11a, chimera profoundly inhibited LFA-1-mediated cell adhesion to ICAM-1, as well as cell spreading following cell adhesion. Unexpectedly, cell adhesion to fibronectin was also inhibited by the CD4/18 chimera. The CD4/18 chimera did not affect the expression of endogenous LFA-1 or the association of CD11a and CD18. Truncation of the carboxyl-terminal 13 amino acid residues of the CD18 cytoplasmic domain of the chimera completely abrogated the inhibitory effect on LFA-1. Among these amino acid residues, the carboxyl-terminal six residues were dispensable for the inhibitory effect in EL-4 cells, whereas it significantly reduced the inhibitory activity of CD4/18 in A20 cells. A larger truncation of the CD18 cytoplasmic domain was needed to fully abrogate the inhibitory effects of CD4/18 on the adhesion to fibronectin. These results show that 1) the CD4/18 chimera has dominant-negative effects on cell adhesion mediated by LFA-1 as well as fibronectin receptors, and 2) amino acid residues of the CD18 cytoplasmic domain involved in the inhibition of LFA-1 seem to be different from those for fibronectin receptors.

Leishmania major: molecular cloning, sequencing, and expression of the heat shock protein 60 gene reveals unique carboxy terminal peptide sequences.

Heat shock proteins (HSP) in the size range of M(r) 60,000 are major targets of the immune response in vivo. The leishmania heat-inducible proteins of M(r) 65-67,000 are expressed at relatively high levels in infected macrophages (Infection and Immunity 1993, 61, 3265-3272) and may be important targets of the host response. To facilitate further studies concerned with these proteins, the HSP60 gene of Leishmania major was cloned, sequenced, and expressed. A lambdaEMBL-3 L. major genomic library was screened with a PCR-generated DNA probe derived from a highly conserved region of the leishmania HSP60 gene. A single clone that hybridized strongly was characterized. Sequence analysis revealed an open reading frame of 1770 bp encoding a putative polypeptide of 589 amino acids with a predicted size of M(r) 64,790 and with the highest degree of amino acid sequence similarity (56%) to HSP60 from Trypanosoma cruzi. Less extensive amino acid sequence similarity (48%) was observed between that leishmania HSP60 and the corresponding human protein. Notably, significant regions of sequence dissimilarity between the leishmania and human proteins were identified principally within the carboxy-terminal regions of the proteins. The entire coding region of the leishmania HSP60 gene was subcloned into the pET-3a vector and expressed in Escherichia coli. Purified recombinant protein was used to examine sera from patients with tegumentary leishmaniasis from Colombia for the presence of antibodies to HSP60. Unlike sera from healthy, uninfected controls, sera from patients reacted strongly with recombinant leishmania HSP60. This recognition had specificity in that these same sera showed little or no reactivity with either recombinant mycobacterial HSP65 or recombinant human HSP60. These findings indicate that patients with tegumentary forms of leishmaniasis have humoral responses to leishmania HSP60. Further studies of this protein will clarify its importance as a target of the immune response and as a potential antigen for serodiagnosis.

Expression of 65- and 67-kilodalton heat-regulated proteins and a 70-kilodalton heat shock cognate protein of Leishmania donovani in macrophages.

Heat shock protein (HSP) expression was examined in murine bone marrow-derived macrophages infected with stationary-phase promastigotes of Leishmania donovani. Immunoblotting performed with a rabbit polyclonal antiserum raised against HSP60 from Heliothis virescens (moth) revealed the de novo appearance of 65- and 67-kDa proteins in leishmania-infected macrophages. A third protein of 60 kDa, which represented murine HSP60, was also detected, and its expression did not change in response to infection. In contrast, expression of the novel 65- and 67-kDa proteins in infected cells was coordinately regulated and, at 24 h of infection, reached maximal levels of 52 to 100% increases above initial levels determined at 3 h. Proteins which had identical electrophoretic mobilities and were similarly regulated in response to heat were also detected in promastigotes. The appearance of these proteins in macrophages was specific to leishmania infection in that neither protein was detected in noninfected cells either in the basal state or following several treatments, including (i) infection with Yersinia pseudotuberculosis, (ii) phagocytosis of Staphylococcus aureus, (iii) NaAsO2 treatment, and (iv) heat shock. Expression of the 65- and 67-kDa heat-regulated Leishmania proteins was also observed to be selective, in that as their concentration was increasing, the abundance of the Leishmania surface protease gp63 in infected cells was noted to decrease. Murine HSP60 but not the Leishmania heat-regulated proteins was also recognized by a distinct rabbit antiserum raised against human HSP60, suggesting the presence of specific determinants within these Leishmania proteins. A monoclonal antibody that recognizes both mammalian HSP70 and HSP70 from plasmodia detected single isoforms of both Leishmania and murine HSP70 in infected cells, and the level of neither protein changed during infection. Moreover, although a murine HSP of 73 kDa was induced in response to both heat shock and NaAsO2 treatment, it was not induced to detectable levels by infection. The rapid and relatively high level of expression of inducible HSP60-related proteins of L. donovani and Leishmania HSP70 in infected macrophages suggests that these proteins are involved in pathogenesis and may be important targets of the immune response.