New vaccines require potent adjuvants like AFPL1 and AFCo1.

Neisseria meningitidis B proteoliposome (AFPL1 when used as adjuvant) and its derivative-Cochleate (AFCo1) contain immunopotentiating and immunomodulating properties and delivery system capacities required for a good adjuvant. Additionally, they contain meningococcal protective antigens and permit packaging of other antigens and pathogen-associated molecular patterns (PAMP). Consequently, we hypothesized that they would function as good vaccine adjuvants for their own antigens and also for non-related antigens. AFPL1 is a detergent-extracted outer membrane vesicle of N. meningitidis B transformed into AFCo1 in calcium environment. Both are produced at Finlay Institute under good manufacture practices (GMP) conditions. We show their exceptional characteristics: combining in the same structure, the potentiator activity, polarizing agents and delivery system capacities; presenting multimeric protein copies; containing multiprotein composition and multi and synergistic PAMP components; acting with incorporated or co-administrated antigens; inducing type I IFN-gamma and IL-12 cytokines suggesting the stimulation of human plasmocytoid precursor and conventional dendritic cells, respectively, inducing a preferential Th1 immune response with TCD4(+), TCD8(+), cross-presentation and cytotoxic T-lymphocyte (CTL) in vivo responses; and functioning by parenteral and mucosal routes. AFPL1-AFCo1 protective protein constitutions permit per se their function as a vaccine. In addition to Phase IV Men BC vaccine, AFPL1 has ended the preclinical stage in an allergy vaccine and is concluding the preclinical stage of a nasal meningococcal vaccine. In conclusion, AFPL1 and AFCo1 induced signal 1, 2 and 3 polarizing to a Th1 (including CTL) response when they acted directly as vaccines or were used as adjuvants with incorporated or co-administered antigens by parenteral or mucosal routes. Both are very promising adjuvants.

Cervicitis actinomicótica: reporte de un caso y revisión de la literatura / Antinomycosis cervix: report of a case and review of the literature

La actinomicosis es una enfermedad infecciosa crónica poco frecuente respecto a su forma clínica más usual. Los casos de actinomicosis que son diagnósticados tardiamente conducen a un manejo inadecuado y a procedimientos quirúrgicos innecesarios. Se reporta un caso de paciente femenina de 46 años de edad con historia clínica de hipermenorrea y anemia. El diagnóstico clínico fue el de miomatosis uterina y el diagnóstico histopatológico fue el de cervicitis por Actinomyces spp. Se hace una revisión de la literatura sobre la incidencia de la actinomicosis en diferentes órganos. Se debe considerar en el diagnóstico diferencial en aquellos casos de uso prolongado de dispositivos intrauterinos, para ayudar a mejorar el manejo y el pronóstico de esta enfermedad

Histiocitosis sinusoidal con adenopatia masiva (enfermedad de rosai-dorfman): presentación de un caso / Sinus histiocytosis with massive lymphadenopathy (disease rosai-dorfman): presentation of a case

La Histiocitosis Sinusoidal con adenopatía masiva o enfermedad de Rosai-Dorfman, se caracteriza por adenopatías cervicales prominentes, bilaterales, dolorosas que se asocian a fiebre, leucocitosis y anemia. La biopsia ganglionar muestra al estudio histológico, fibrosis capsular y sinusoides dilatados ocupados por histiocitos, los cuales suelen mostrar emperipolesis, como carcaterística fundamental. Clínicamente tiene curso indolente con desaparición espontánea de las adenomegalias. Se presenta el caso de un perescolar masculino de 5 años con clínica de adenomegalias cervicales bilaterales

Anticuerpos IgA anti-lipopolisacárido inducidos por candidato vacunal oral 638 vivo / Mucosal IgA anti-lipopolysaccharide antibodies induced by 638 oral live attenuated candidate vaccine

La cepa de Cólera atenuada 638 ha inducido una buena respuesta en modelos animales y en un estudio piloto humano ha probado ser segura e inmunogénica. Sin embargo, no ha sido evaluada la IgA específica en mucosas ni tampoco se ha comparado la respuesta inducida por la cepa 638 con aquélla inducida por la conocida cepa reactogénica JBK70. Por ello, fueron evaluadas las células secretoras de anticuerpos (ASC) anti-lipopolisacárido (LPS) sanguíneas y los anticuerpos antiLPS en saliva como indicadores de respuestas mucosas de voluntarios inoculados con las cepas 638 o JBK70. Con vistas a determinar la producción local o no de la IgA específica, la cinética de los anticuerpos IgA anti-LPS séricos y salivares fueron comparados. La respuesta vibriocida sérica fue también medida. Tres grupos con 638 (109, 108 y 107 unidades form adoras de colonias, CFU), uno con JBK70 (109 CFU) y otro con placebo fueron enrolados. La respuesta sérica de ASC IgA+ fue mayor que la de ASC IgG+.La IgA anti-LPS en saliva tuvo valores máximos a los 9 días y decayó hasta valores negativos en el día 14 después de la inoculación. La IgA anti-LPS sérica permanece elevada entre los 7 y 28 días después de la inoculación lo que sugiere que la IgA en saliva es localmente y transitoriamente producida. La respuesta vibriocida sérica fue incrementada después de la inoculación. Respuestas similares fueron obtenidas con las cepas 638 y JBK70 (AU)

Immune response induction and new effector mechanisms possibly involved in protection conferred by the Cuban anti-meningococcal BC vaccine.

This report explores the participation of some afferent mechanisms in the immune response induced by the Cuban anti-meningococcal vaccine VA-MENGOC-BC. The induction of delayed-type hypersensitivity in nursing babies and lymphocyte proliferation after immunization is demonstrated. The presence of gamma interferon IFN-gamma and interleukin-2 (IL-2) mRNAs but absence of IL-4, IL-5, and IL-10 mRNAs were observed in peripheral blood mononuclear cells from immunized subjects after in vitro challenge with outer membrane vesicles. In addition, some effector functions were also explored. The presence of opsonic activity was demonstrated in sera from vaccinees. The role of neutrophils as essential effector cells was shown. In conclusion, we have shown that, at least in the Cuban adult population, VA-MENGOC-BC induces mechanisms with a T-helper 1 pattern in the afferent and effector branches of the immune response.

Síndrome nefrótico congénito y síndrome de down: reporte de un caso y revisión de la literatura / Congenital nephrotic syndrome and down syndrome: report of case and literature revision

El síndrome nefrótico congénito durante el primer año de vida es poco frecuente en nuestro medio, al igual que en otros países de latinoamérica; lo contrario ocurre en otros países europeos donde la prevalencia es mayor: Se ha encontrado asociaciones de esta síndrome con alteraciones embrionarias como el nefroblastoma, anormalidades genitales, seudohermafroditismo, síndrome uñarótula y displasia congénita de la cadera. Se describe el caso clínico de un lactante menor femenina de 6 meses de edad con síndrome nefrótico congénito y síndrome de down; esta asociación no ha sido descrita previamente en la literatura nacional. El estudio histopatológico mostró disminución en el número de glomerulos y la lesión observada en los gromérulos fue una esclerosis mesangial difusa retráctil

Microgravity alters protein phosphorylation changes during initiation of sea urchin sperm motility.

European Space Agency (ESA) studies demonstrated that bull sperm swim with higher velocity in microgravity (microG) than at 1 G. Coupling between protein phosphorylation and sperm motility during activation in microG and at 1 G was examined in the ESA Biorack on two space shuttle missions. Immotile sperm were activated to swim (86-90% motility) at launch +20 h by dilution into artificial seawater (ASW). Parallel ground controls were performed 2 h after the flight experiment. Activation after 0, 30, and 60 s was terminated with electrophoresis sample buffer and samples analyzed for phosphoamino acids by Western blotting. Phosphorylation of a 130-kDa phosphothreonine-containing protein (FP130) occurred three to four times faster in microG than at 1 G. A 32-kDa phosphoserine-containing protein was significantly stimulated at 30 s but returned to 1 G control levels at 60 s. The rate of FP130 phosphorylation in microG was attenuated by D2O, suggesting that changes in water properties participate in altering signal transduction. Changes in FP130 phosphorylation triggered by the egg peptide speract were delayed in microG. These results demonstrate that previously observed effects of microG on sperm motility are coupled to changes in phosphorylation of specific flagellar proteins and that early events of sperm activation and fertilization are altered in microG.

Fibroma cardíaco / Cardiac fibroma

Se reporta un caso de Fibroma cardíaco en un lactante masculino menor, de siete meses de edad. El tumor estaba localizado en la cara diafragmática, bien circunscrita y firme. Hispatológicamente el tumor está constituido por tejido conectivo fibroso y colágeno entremezclado sin actividad mitótica. La coloración de tricómico de Gomori demostró su origen fibroblástico

Identification of phosphoproteins coupled to initiation of motility in live epididymal mouse sperm.

A method for collecting live immotile cauda epididymal mouse sperm that initiate motility by dilution into an activation buffer is described. Sperm in collection buffer showed low percent motility (MOT) and population progression (PRG) that increased 10-fold and 9-fold, respectively, during the first 2 min after dilution into activation buffer. Western phosphoserine (pS), phosphothreonine (pT), and phosphotyrosine (pY) analysis revealed a 120 kDa protein that markedly increased in pT content during initiation of motility and may be related to FP130, the motility-coupled axonemal protein of sea urchin sperm. A prominent 82 kDa protein that was pS and pT-phosphorylated in immotile and motile sperm is likely the fibrous sheath component AKAP82 that is phosphorylated during spermatogenesis. Analysis of live human sperm also identified a prominent 120 kDa pT protein. Thus it appears that phosphorylation of FP130 and related 120 kDa proteins in mouse, and perhaps human sperm, represent common targets during motility initiation in sperm.

Identification of flagellar proteins that initiate the activation of sperm motility in vivo.

Protein phosphorylation appears to be a necessary step in the intracellular signaling pathway that initiates the activation of sperm motility. Activation of live immotile sea urchin sperm produced rapid, time-dependent increased phosphorylation on proteins of 32, 45, 130, and 500 kDa. Fractionation of immotile and motile sperm indicated that these motility-related phosphoproteins are associated with flagella. These proteins showed greater phosphorylation in the flagellar fraction from motile sperm, suggesting that subcellular boundaries are in place to keep protein kinases and their substrates spatially separated. Solubility properties suggest that these proteins are the heavy chain and smaller subunits of sea urchin sperm dynein which are phosphorylated in vivo to initiate activation of motility. This also suggests that phosphorylation of only these few proteins, out of the nearly 100 phosphorylations known to occur in the basic axoneme, appears to be associated with the early signaling pathways of motility activation in intact sperm.

A method for preparation, storage, and activation of large populations of immotile sea urchin sperm.

Reversible protein phosphorylation is associated with initiation and modulation of sperm flagellar motility. Many studies aimed at examining the signal transduction mechanisms underlying the expression of motility have relied on detergent-permeabilized sperm reactivated with exogenous 32P-ATP. However, the reactivation conditions allow variable levels of motility to be expressed and phosphorylation of many proteins that appear to be unrelated to sperm motility. Thus, identification of the few relevant proteins is difficult. We have developed a method to collect and keep sperm immotile until reactivated for analysis to normal motility levels. Artificial sea water (ASW) buffered with 5 mM 2-[N-morpholino]ethanesulfonic acid at pH 6.0 and containing 50 mM KCl allows collection and storage of immotile sea urchin sperm for up to 96 h at 4-5 degrees C. Motility under these conditions is essentially zero, but sperm is rapidly reactivated to normal motility by diluting with ASW to standard pH (8.0) and KCl concentration (10 mM).

Regulation of human sperm motility and hyperactivation components by calcium, calmodulin, and protein phosphatases.

The role of Ca2+, calmodulin, and protein phosphatases on motility and hyperactivation of noncapacitated, capacitating, and detergent-permeabilized reactivated human sperm was examined. In noncapacitated sperm, W7 inhibited percent motility (%MOT), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), and percent hyperactivation (%HYP) in an extracellular Ca2+ concentration-dependent manner (p < .05). However, in capacitating sperm, inhibition of motility by W7 was independent of external Ca2+. Treatment of reactivated sperm with a synthetic calmodulin inhibitor peptide decreased VCL and ALH in a Ca(2+)-dependent manner (p < .05). Ca2+ exhibited a dramatic influence on motility within a narrow concentration range (0.7 to 1.0 microM) in reactivated sperm. A calmodulin-dependent protein phosphatase (PP2B) was identified by activity assay, immunoblotting, and dephosphorylation of endogenous phosphoproteins. The sperm enzyme, unlike bovine brain PP2B, was inhibited by 1 microM okadaic acid (OA) in the presence of Mn2+, suggesting that the sperm enzyme is unique. In reactivated sperm, inhibition of endogenous PP2B-like activity with anti-PP2B antibodies altered ALH, whereas OA altered both VCL and ALH and also inhibited a subset of Ca(2+)-dependent dephosphorylations of cAMP-dependent phosphoproteins in capacitating sperm. These results suggest (1) an important role for calmodulin and PP2B in Ca(2+)-regulated motility parameters, particularly ALH, and (2) that modulation of human sperm motility, including hyperactivation by cAMP-dependent phosphorylation, requires calmodulin-dependent as well as other protein dephosphorylations.

Characteristics of the Inhibition of Potato (Solanum tuberosum) Invertase by an Endogenous Proteinaceous Inhibitor in Potatoes.

Effect of several parameters on inhibition of potato (Solanum tuberosum) invertase by its endogenous proteinaceous inhibitor was determined using homogeneous preparations of both proteins. The inhibitor and invertase formed an inactive complex with an observed association rate constant at pH 4.70 and 37 degrees C of 8.82 x 10(2) per molar per second and a dissociation rate constant of 3.3 x 10(-3) per minute. The inhibitor appeared to bind to invertase in more than one step. Initial interaction (measured by loss of invertase activity) was rapid, relatively weak, readily reversible (K(i) of 2 x 10(-6) molar) and noncompetitive with substrate at pH 4.70. Initial interaction was probably followed by isomerization to a tighter (K(i) of 6.23 x 10(-8) molar) complex, which dissociated slowly with a half-time of 3.5 hour. Interaction between enzyme and inhibitor appeared to be of ionic character and essentially pH independent between pH 3.5 and 7.4.

Purification and Partial Characterization of Potato (Solanum tuberosum) Invertase and Its Endogenous Proteinaceous Inhibitor.

Invertase plays an important role in the hydrolysis of sucrose in higher plants, especially in the storage organs. In potato (Solanum tuberosum) tubers, and in some other plant tissues, the enzyme seems to be controlled by interaction with an endogenous proteinaceous inhibitor. An acid invertase from potato tubers (variety russet) was purified 1560-fold to electrophoretic homogeneity by consecutive use of concanvalin A-Sepharose 4B affinity chromatography, DEAE-Sephadex A-50-120 chromatography, Sephadex G-150 chromatography, and DEAE-Sephadex A-50-120 chromatography. The enzyme contained 10.9% carbohydrate, had an apparent molecular weight of 60,000 by gel filtration, and was composed of two identical molecular weight subunits (M(r) 30,000). The enzyme had a K(m) for sucrose of 16 millimolar at pH 4.70 and was most stable and had maximum activity around pH 5. The endogenous inhibitor was purified 610-fold to homogeneity by consecutive treatment at pH 1 to 1.5 at 37 degrees C for 1 hour, (NH(4))(2)SO(4) fractionation, Sephadex G-100 chromatography, DEAE-Sephadex G-50-120 chromatography, and hydroxylapatite chromatography. The inhibitor appears to be a single polypeptide (M(r) 17,000) without glyco groups. The purified inhibitor was stable over the pH range of 2 to 7 when incubated at 37 degrees C for 1 hour.