Adjuvant requirement for successful immunization with recombinant derivatives of Plasmodium vivax merozoite surface protein-1 delivered via the intranasal route

Recently, we generated two bacterial recombinant proteins expressing 89 amino acids of the C-terminal domain of the Plasmodium vivax merozoite surface protein-1 and the hexa-histidine tag (His6MSP1(19)). One of these recombinant proteins contained also the amino acid sequence of the universal pan allelic T-cell epitope (His6MSP1(19)-PADRE). In the present study, we evaluated the immunogenic properties of these antigens when administered via the intra-nasal route in the presence of distinct adjuvant formulations. We found that C57BL/6 mice immunized with either recombinant proteins in the presence of the adjuvants cholera toxin (CT) or the Escherichia coli heat labile toxin (LT) developed high and long lasting titers of specific serum antibodies. The induced immune responses reached maximum levels after three immunizing doses with a prevailing IgG1 subclass response. In contrast, mice immunized by intranasal route with His6MSP1(19)-PADRE in the presence of the synthetic oligonucleotides adjuvant CpG ODN 1826 developed lower antibody titers but when combined to CT, CpG addition resulted in enhanced IgG responses characterized by lower IgG1 levels. Considering the limitations of antigens formulations that can be used in humans, mucosal adjuvants can be a reliable alternative for the development of new strategies of immunization using recombinant proteins of P. vivax.

In vitro compatibility of remifentanil hydrochloride and sufentanil citrate with selected drugs

: Objectives Physicochemical incompatibilities between intravenous drugs are a recurrent problem in intensive care units. The present study was aimed at investigating the physical compatibility of remifentanil and sufentanil with other drugs (insulin, midazolam, propofol, potassium chloride, magnesium sulfate, furosemide, heparin, monobasic potassium phosphate) that are frequently administered together intravenously. In addition, the physicochemical compatibility of three common associations of drugs was evaluated in glass tube tests and during dynamic simulated Y site administrations (remifentanil-insulin-midazolam; remifentanil-insulin-propofol; sufentanil-insulin-midazolam). Methods Physical compatibility was verified by visual inspection of the various mixtures (two, three or four drugs) in glass tubes and by pH determination of the mixtures collected during simulated Y site administrations. Solutions were considered as compatible in the absence of any visual change in the solution and of any significant variation in pH value. In addition, chemical stability was checked during in vitro dynamic simulations. The solutions were prepared in 50 ml syringes, placed on syringe pumps and connected to a Swan-Ganz catheter; the liquid collected at the tip was assayed by high performance liquid chromatography. Results In the visual examinations, only the associations of remifentanil and furosemide were incompatible. The three assayed associations were compatible in the tested proportion range over 24 h. Conclusions Remifentanil was physically compatible with the tested drugs, except for furosemide (Lasix; Sanofi-Aventis, 250 mg/25 ml) and physicochemically compatible with insulin and midazolam and insulin and propofol. Sufentanil was physically compatible with all tested drugs and physicochemically compatible with insulin and midazolam

Intranasal administration of the synthetic polypeptide from the C-terminus of the circumsporozoite protein of Plasmodium berghei with the modified heat-labile toxin of Escherichia coli (LTK63) induces a complete protection against malaria challenge.

Needle-free procedures are very attractive ways to deliver vaccines because they diminish the risk of contamination and may reduce local reactions, pain or pain fear especially in young children with a consequence of increasing the vaccination coverage for the whole population. For this purpose, the possible development of a mucosal malaria vaccine was investigated. Intranasal immunization was performed in BALB/c mice using a well-studied Plasmodium berghei model antigen derived from the circumsporozoite protein with the modified heat-labile toxin of Escherichia coli (LTK63), which is devoid of any enzymatic activity compared to the wild type form. Here, we show that intranasal administration of the two compounds activates the T and B cell immune response locally and systemically. In addition, a total protection of mice is obtained upon a challenge with live sporozoites.

Effects of intranasal administration of synthetic (4-28)human atrial natriuretic peptide to normal volunteers.

The effects of intranasal administration of increasing doses of synthetic human natriuretic peptide (4-28 hANP) were studied in six healthy volunteers. The peptide was administered as a nasal spray at doses of 50, 100, 200, and 500 micrograms in ascending order at 48-h intervals. Vehicle was administered by the same route randomly between any two of the doses. Intranasal hANP administration had no effect on either blood pressure, heart rate (HR), or hematocrit. Diuresis did not change consistently, whereas natriuresis tended to rise with vehicle as well as with hANP administration. This was attributed to the infusion of isotonic saline during the experiment. There was no significant increase in plasma ANP levels after intranasal administration of any of the different doses. Thus, no evidence that the atrial natriuretic peptide tested (4-28 hANP) can cross the nasal mucosal barrier was found.

Intragastric and Intranasal Administration of Lactobacillus paracasei NCC2461 Modulates Allergic Airway Inflammation in Mice.

Introduction. Preclinical and clinical evidences for a role of oral probiotics in the management of allergic diseases are emerging. Aim. We aimed at testing the immunomodulatory effects of intranasal versus intragastric administration of Lactobacillus paracasei NCC2461 in a mouse model of allergic airway inflammation and the specificity of different probiotics by comparing L. paracasei NCC2461 to Lactobacillus plantarum NCC1107. Methods. L. paracasei NCC2461 or L. plantarum NCC1107 strains were administered either intragastrically (NCC2461) or intranasally (NCC2461 or NCC1107) to OVA-sensitized mice challenged with OVA aerosols. Inflammatory cell recruitment into BALF, eotaxin and IL-5 production in the lungs were measured. Results. Intranasal L. paracasei NCC2461 efficiently protected sensitized mice upon exposure to OVA aerosols in a dose-dependent manner as compared to control mice. Inflammatory cell number, eotaxin and IL-5 were significantly reduced in BALF. Intranasal supplementation of L. paracasei NCC2461 was more potent than intragastric application in limiting the allergic response and possibly linked to an increase in T regulatory cells in the lungs. Finally, intranasal L. plantarum NCC1107 reduced total and eosinophilic lung inflammation, but increased neutrophilia and macrophages infiltration. Conclusion. A concerted selection of intervention schedule, doses, and administration routes (intranasal versus intragastric) may markedly contribute to modulate airway inflammation in a probiotic strain-specific manner.

Severe pneumonia due to Parachlamydia acanthamoebae following intranasal inoculation: a mice model.

Parachlamydia acanthamoebae is an obligate intracellular bacterium naturally infecting free-living amoebae. The role of this bacterium as an agent of pneumonia is suggested by sero-epidemiological studies and molecular surveys. Furthermore, P. acanthamoebae may escape macrophages microbicidal effectors. Recently, we demonstrated that intratracheal inoculation of P. acanthamoebae induced pneumonia in 100% of infected mice. However, the intratracheal route of infection is not the natural way of infection and we therefore developed an intranasal murine model. Mice inoculated with P. acanthamoebae by intranasal inoculation lost 18% of their weight up to 8 days post-inoculation. All mice presented histological signs of pneumonia at day 2, 4, 7, and 10 post-inoculation, whereas no control mice harboured signs of pneumonia. A 5-fold increase in bacterial load was observed from day 0 to day 4 post-inoculation. Lungs of inoculated mice were positive by Parachlamydia-specific immunohistochemistry 4 days post-inoculation, and P. acanthamoebae were localized within macrophages. Thus, we demonstrated that P. acanthamoebae induce a severe pneumonia in mice. This animal model (i) further supports the role of P. acanthamoebae as an agent of pneumonia, confirming the third Koch postulate, and (ii) identified alveolar macrophages as one of the initial cells where P. acanthamoebae is localized following infection.

Efeitos anestésicos da administração intranasal ou intramuscular de cetamina S+ e midazolam em pomba-rola (Streptotelia sp.)

A via intranasal é uma boa alternativa por ser indolor e de fácil aplicação em aves. O objetivo deste estudo foi avaliar os efeitos anestésicos da associação de cetamina S+ e midazolam pela via intranasal (IN) em comparação com a via intramuscular (IM) em pombos. Foram utilizados 12 pombos alocados em dois grupos com 15 dias de intervalo, os quais receberam: grupo IM: 20 mg/kg de cetamina S+ associada a 3,5 mg/kg de midazolam pela via intramuscular (musculatura do peito); e grupo IN, mesmo protocolo, porém, pela via intranasal. Os parâmetros avaliados foram: período de latência, tempo de duração em decúbito dorsal, tempo total de anestesia, tempo de recuperação e efeitos adversos. Para a análise estatística, empregou-se o teste de Wilcoxon, com as diferenças consideradas significativas quando P<0,05. O período de latência obtido foi de 30 [30-47,5] e 40 [30-50] segundos para IM e IN, respectivamente. O tempo de duração de decúbito dorsal foi de 59 [53,25-65] e 63 [37-71,25] minutos para IM e IN, respectivamente, sem diferenças significativas entre os grupos. Com relação à duração total de anestesia, foi observada diferença significativa, com 88 [86,25-94,5] e 68 [53,5-93] minutos para os grupos IM e IN, respectivamente. O tempo de recuperação foi mais curto no grupo IN (15 [4,25-19,5]) comparado ao IM (32 [28,25-38,25] minutos). Dois animais de cada grupo apresentaram regurgitação na fase de recuperação. Conclui-se que a administração de cetamina S+ e midazolam pela via intranasal é um método aceitável de administração de fármacos e produz anestesia rápida e eficaz em pombos.

Efeitos sedativos da associação de Cetamina e Midazolam administrados pela via intranasal ou intramuscular em papagaio (Amazona aestiva e Amazona vinacea)

A falta de protocolos de sedação seguros para uso em papagaios na literatura demonstra a necessidade de conhecer os anestésicos que são eficazes nestes animais. Devido a pouca massa muscular desta espécie, notou-se a necessidade de estudar outra via de administração, menos invasiva e dolorosa ao animal, como a via intranasal. O objetivo deste estudo foi avaliar os efeitos sedativos e a viabilidade da administração intranasal, em comparação à via intramuscular, de 15mg/kg de Cetamina e 1mg/kg de Midazolam. Foram utilizados 14 papagaios das espécies Amazona aestiva e Amazona vinacea, de ambos os sexos, adultos, peso médio de 388,5±29,1g. Os animais foram distribuídos aleatoriamente em dois grupos: intramuscular (IM, n=7) e intranasal (IN, n=7). No grupo intramuscular, a administração dos anestésicos foi realizada nos músculos peitorais, utilizando seringas de insulina e no grupo intranasal, com auxílio de uma micropipeta. Avaliou-se o período de latência, tempo de duração, qualidade de sedação, e o tempo de recuperação total. A média para o período de latência no grupo IM foi de 6,13±2,02 minutos e no grupo IN de 4,84±2,37 minutos. Já para o tempo de duração da sedação no grupo IM a média foi de 35,81±29,56 e no grupo IN de 24,52±14,83 minutos. Ambas as vias promoveram sedação adequada, pois a média do escore da qualidade de sedação obtida pelo grupo IM foi 2±1,5 e pelo grupo IN 1,28±1,1. O tempo de recuperação total no grupo IM foi de 27,04±11,69 e no grupo IN de 17,67±11,64 minutos. Apesar do grupo IN ter apresentado os menores tempos de período de latência, duração e de recuperação total e ter obtido melhor escore na qualidade de sedação, não houve diferença estatística significativa entre os grupos. Os resultados obtidos neste estudo indicam que a administração de 15 mg/kg de cetamina e 1mg/kg de midazolam pela via intranasal ou intramuscular em papagaios (Amazona aestiva e Amazona vinacea) produzem sedação adequada para pequenos procedimentos como colocação de anilha, coleta de sangue e radiografias; porém a via intranasal mostrou ser uma alternativa menos invasiva quando comparado à via intramuscular.

Impacto en la calidad de la Inducción anestésica con premedicación en pacientes pediatricos

La inducción anestésica en niños es uno de los mayores retos para los anestesiólogos ya que es la fase más estresante del período perioperatorio para el paciente pediátrico. Existen diferentes intervenciones para minimizar la ansiedad perioperatoria y aumentar la cooperación del paciente pediátrico con la inducción anestésica. Entre las intervenciones exitosas la premedicación farmacológica con midazolam ha mostrado grandes beneficios en pacientes pediátricos. Metodología: Se realizó un estudio de casos y controles en pacientes pediátricos llevados a cirugía en la Fundación Cardioinfantil entre 2011-2014. Por medio de muestreo aleatorio por conveniencia se tomaron como casos pacientes con premedicación y controles pacientes sin premedicación. El éxito en la inducción se midió por medio de la escala ICC, usada a nivel mundial. Resultados: El promedio de edad fue 4.9 σ 3.01 años para los casos y 5.02σ3.2 años para controles, presentaron la misma distribución por género, 40.6% femenino, 59.3% masculino. El éxito de la inducción anestésica con midazolam mostró resultados significativos (OR 7.3 IC95% 4.3 – 12.5 p0,000), en hombres (OR 9.44 IC95%4.5 – 19.8 p0,000), en menores de 5 años (OR 10.33 IC95% 5.07 – 21.04 p0,000), en pacientes con antecedentes quirúrgicos (OR 12.2 IC95% 5.28 – 27.8 p0.000) o anestesias previas (OR 7.9 IC95% 4.4 – 14.4 p0,000). Discusión: El midazolam como agente farmacológico usado para premedicación en pacientes pediátricos presenta resultados exitosos contundentes, por lo cual debe usarse en todos los casos.

Early treatment with intranasal neostigmine reduces mortality in a mouse model of Naja naja (Indian Cobra) envenomation

Objective. Most snakebite deaths occur prior to hospital arrival; yet inexpensive, effective, and easy to administer out-of-hospital treatments do not exist. Acetylcholinesterase inhibitors can be therapeutic in neurotoxic envenomations when administered intravenously, but nasally delivered drugs could facilitate prehospital therapy for these patients. We tested the feasibility of this idea in experimentally envenomed mice. Methods. Mice received intraperitoneal injections of Naja naja venom 2.5 to 10 times the estimated LD50 and then received 5

Toxoplasma gondii: Evaluation of an intranasal vaccine using recombinant proteins against brain cyst formation in BALB/c mice

The purpose of this work was to evaluate protective activity against brain cyst formation in BALB/c mice intranasally vaccinated with recombinant proteins from Toxoplasma gondii. The recombinant proteins rROP2, rGRA5 and rGRA7 were used in vaccine preparation. Thirty-three female mice were divided into three groups, these animals received two doses by intranasal route at days 0 and 21 as follows; group 1 (G1, n = 11) received 12.5 mu g of each recombinant protein plus 0.5 mu g of cholera toxin, group 2 (G2, n = 11) received phosphate buffer saline (PBS) plus 0.5 mu g of cholera toxin, and group 3 (G3, n = 11) received PBS only. At challenge day (day 33) three animals from each group were euthanatized for IgA measure from intestine. Mice were infected orally with 50 cysts from the VEG strain at day 33. At challenge day the G1 animals had high immunoglobulin A levels, however, they only showed IgG antibody titers against rROP2 and rGRAT Animals from G1 also exhibited strong resistance to cyst formation compared with the control group (G3, P < 0.05). However, we did not observe differences in protection against brain cyst formation between G1 and G2 (P > 0.1). These results indicate that intranasal immunization in BALB/c mice with recombinant proteins rROP2, rGRA5 and rGRA7 associated with cholera toxin induced partial protection, when compared with G3, against tissue cyst formation after oral infection with tissue cysts from T gondii. (c) 2007 Elsevier B.V. All rights reserved.

Intranasal delivery of zidovudine by PLA and PLA-PEG blend nanoparticles

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Reversed phase HPLC determination of zidovudine in rat plasma and its pharmacokinetics after a single intranasal dose administration

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)