Transfection improvements of fish cell lines by using deacylated polyethylenimine of selected molecular weights.

A new tool for DNA transfer to fish cell lines such as epithelioma papulosum cyprini (EPC) and rainbow trout gonad (RTG2), has been optimized by testing commercially available polyethylenimine (PEI) polymers as transfectant reagents. Deacylated 25 kDa PEI polymers were selected amongst the most active and then low toxicity deacylated PEIs fractions around 15 kDa were obtained by gel filtration chromatography to increase 3-4-fold their initial in vitro transfection efficiency. The EPC and plasmids coding for reporter genes were first used to optimize variable values for best expression by transfection with deacylated low toxicity PEI while both EPC/RTG2 and a plasmid coding for the glycoprotein G gene of the fish pathogen, viral haemorrhagic septicemia virus (VHSV) were then used to demonstrate some of their practical applications. Due to its relatively low price, defined chemical composition and availability, low toxicity deacylated PEI might be used for numerous applications for all those studying fish cell immunology in vitro as well as in vivo.

[Plasmatic concentrations of fibronectin as marker of clinical course among septic patients]. / Concentraciones plasmáticas de fibronectina como marcador de evolución en pacientes sépticos.

INTRODUCTION: It is difficult to follow up the patients with sepsis because of the underlying pathology or the presence of complications. The attending physician needs to know as soon as possible the response to the antibiotic therapy. Is therefore necessary to find real time biological markers that will help to understand the clinical situation of the infected patient. It has been demonstrated that fibronectin (Fn) is an early marker of sepsis, so it seemed plausible its use in the clinical monitoring of the septic patient. MATERIAL: We have used plasmatic concentrations of Fn in the follow up of sepic patients admitted to the Hospital de Segovia from February to August, 1995. There were two control groups: one of healthy volunteers and another of varied pathology. RESULTS: We have applied the multiple comparison rule of Bonferroni to demonstrate that plasma Fn is a negative acute phase reactant, with almost undetectable levels while the patient is in critical condition. An effective treatment prompts plasma Fn to rise significantly within two days. DISCUSSION: Plasma Fn concentration by itself or in an score system could help in the follow up of the septic patients.

Report of six cases of human infection by Serratia plymuthica.

Serratia plymuthica is an uncommon cause of human infection. Only one case of chronic osteomyelitis and two cases of sepsis secondary to central venous catheter infection have been documented. We report the isolation of S. plymuthica from six patients. The organism was recovered from blood cultures in three cases in which the patients had lymphoblastic leukemia, lymphoma, or stroke. Two isolates were recovered from exudates (following knee and abdominal surgery). In the last case, the organism was isolated from the peritoneal fluid of a patient with cholecystitis. The infection was considered nosocomial in five cases and community acquired in the other.

[Brucellosis after treatment: the diagnosis of recurrences]. / La brucelosis después del tratamiento: diagnóstico de las recidivas.

BACKGROUND: Given the frequency of relapse following treatment of brucellosis as well as the persistence of symptoms the authors attempted to discover a recommendable position for surveillance of patients once treatment has been completed. METHODS: A series of 105 patients treated for brucellosis was prospectively analyzed, 11 of whom had relapse. Post treatment symptoms, serology titers and the results of the cultures between the cured patients and those with relapse were compared. RESULTS: Hemocultures (sensitivity 0.73) and "specific" symptoms (prolonged fever, spondylitis, arthritis, orchitis and others) were the most useful data for diagnosis of relapse. The presence of vague symptoms following treatment was of slight value specially in the first months. Elevation of serology titers following treatment offered little aid in diagnosis (likelihood ratio 1.7). The persistence of high titers of serology several months following completion of treatment offered high likelihood ratios which may aid in therapeutic decision making. CONCLUSIONS: It is recommended that no tests be made on asymptomatic patients following treatment of brucellosis and to treat those patients with "specific" symptoms. Hemocultures are recommended for patients with vague symptoms and if several months have passed serology should be carried out from which the probability of relapse may be calculated.

[Diagnosis of brucellosis in an endemic area. Evaluation of routine diagnostic tests]. / El diagnóstico de la brucelosis en un área endémica. Valoración de las pruebas diagnósticas habituales.

BACKGROUND: The use of classic methods of diagnosis of brucellosis was analyzed, particularly serologic methods whose use in endemic areas and risk groups has been questioned in the literature. METHODS: Prospective analysis of these methods was performed in a group of 171 patients suspected as having brucellosis proceeding from an endemic area, with progressions of risk and frequent antecedents of brucellosis. The results obtained were compared in 119 patients in whom brucellosis was confirmed (80 with positive cultures and 39 with clinic-serologic diagnosis) and in 52 in whom the diagnosis was excluded. RESULTS: The hemocultures provided a sensitivity of 70% with a mean delay in growth of 13.6 days. The rise of Bengal test showed sensitivity of 95% and specificity of 75%. The most adequate cut-off point for seroagglutination was of 1/160 and for the Coombs test 1/320 with sensitivities of 93 and 92% and specificities of 97 and 100%, respectively. CONCLUSIONS: The routine serologic tests offer good results for the diagnosis of brucellosis in endemic areas upon use of adequate cut-off points and permitting therapeutic decisions to be taken prior to knowledge of the results of the cultures. The rose of Bengal test is valid for initial selection of this type of population.

Sacral osteomyelitis due to Nocardia asteroides. A case report.

Nocardial osteomyelitis is increasing in both immunocompetent and immunosuppressed patients. We report a case of a Nocardia asteroides infection of the sacrum in a 37-year-old man who was successfully treated surgically.