Partitioning of whey proteins, bovine serum albumin and porcine insulin in aqueous two-phase systems

Partitioning of the proteins from cheese whey, bovine serum albumin and porcine insulin were analysed using aqueous two-phase systems (ATPS) prepared with PEG-phosphate, PEG-citrate and PEG-maltodextrin (MD). Proteins were quantified through one of the following methods: FPLC, Bradford and spectrophotometry at 280 nm. Results showed that whey proteins partitioned unevenly on the phases of the systems used, with alpha-lactoalbumin (alpha-La) concentrated in the upper phase and beta-lactoglobulin (beta-Lg) in the lower. Albumin in PEG-MD systems concentrated in the MD-rich lower phase. Porcine insulin showed great affinity with the PEG-rich phase, its partition coefficient was always over 10 and increases with PEG molecular mass.     

Indoor mite allergens in patients with respiratory allergy living in Porto, Portugal

We investigated the levels of mite allergens (Der p 1, Der f 1, Der 2, and Lep d 1) in dust samples from the homes of 59 patients with asthma, 36 sensitized to house-dust mites (HDM) and 23 to grass pollen (controls), living in Porto, northern Portugal. The relationship between exposure and sensitization to HDM and the influence of housing conditions on mite-allergen levels were also evaluated. Der p 1 (median 9.2 μg/g) and Der 2 (4.6 μg/g) were the main allergens, while Der f 1 and Lep d 1 levels were always <1 μg/g dust and undetectable in 11% and 47% of samples, respectively. All HDM-sensitized asthmatics were exposed to Der p 1 levels >2 μg/g and their homes contained significantly higher levels of Der p 1 (median 12.5 vs 6.4 μg/g; P=0.008) and Der 2 (6.2 vs 3.0 μg/g; P=0.004) when compared to the control group. A significant correlation was observed between the exposure to Der p 1 and the wheal area at skin testing with the Dermatophagoides pteronyssinus (Dp) extract (P=0.01) as well as with serum specific IgE levels to Dp (P=0.03). Patients with higher levels of serum specific IgE (≥17.5 HRU/ml) were also more frequently exposed to Der p 1 levels ≥10 (μg/g (P=0.002). Old homes, presence of carpets, and signs of dampness were conditions associated with significantly higher levels of mite allergens. In conclusion, we found high levels of Der p 1 and Der 2 particularly in the homes of HDM-sensitized patients and we confirm the relationship between exposure and sensitization to HDM, assessed by both in vivo and in vitro methods. In addition to a favorable outdoor climate, we found in our region housing conditions propitious to mite growth, suggesting that specific geographic characteristics must also be taken into account for the correct planning of mite-avoidance measures.     

Plasmid profiles of "Campylobacter upsaliensis" isolated from blood cultures and stools of paediatric patients.

Seventy-three clinical isolates of "Campylobacter upsaliensis" were screened for the presence of plasmids. Plasmid bands were found in 68 (93%) isolates, from which 14 plasmid types were identified. Type 5 was found only in blood-culture isolates, whereas types 7-14 were found only in faecal isolates. Plasmid-free isolates and the other plasmid profiles were present in both faecal and blood isolates. The reproducibility of these profiles was largely dependent on the method of plasmid isolation. The chloroform-phenol lysis method was the most efficient and reliable method of preparing plasmid DNA for plasmid profile analysis. The success of this method may be largely attributable to two factors: (1) the efficacy of cell lysis was independent of either an alkaline agent or of heat; (2) the inactivation of nucleases by the utilisation of chloroform-phenol to lyse the cells. Furthermore, this method of plasmid DNA preparation is ideally suited for use on clinical isolates, especially when rapid plasmid profile analysis may be required.     

Creatine supplementation and riluzole treatment provide similar beneficial effects in copper,zinc superoxide dismutase (G93A) transgenic mice

This study investigated the effects of riluzole (Ril), creatine (Cr) and a combination of these treatments on the onset and progression of clinical signs and neuropathology in an animal model of familial amyotrophic lateral sclerosis, the G93A transgenic mouse (n=13-17 per group). The onset of clinical signs was delayed (P<0.05) by about 12 days in all treatment groups compared with control; however, no differences occurred between treatments. All animals were killed at 199 days of age. At the end of the experimental period the severity of clinical signs was less (P<0.05) with all treatments compared with control. Again no differences between treatments were observed. The treatments had no effect on the number of neurons in ventral horns of the lumbar region of the spinal cord. Transgenic mice ingesting Cr displayed elevated (P<0.05) total Cr levels in cerebral hemispheres (5%) and spinal cord (8%), but not skeletal muscles. These data demonstrate that treatment with Ril and Cr were both effective in delaying disease onset and clinical disability. To the age of killing, no additional benefit was conferred by co-administration of Ril and Cr.     

Sertoli-Leydig cell tumor of the ovary with alpha-fetoprotein production

A case of poorly differentiated Sertoli-Leydig cell tumor with elevated serum alpha-fetoprotein (AFP) levels occurred in a 17-year-old girl. No germ cell component and no heterologous elements were identified, but a retiform pattern was present. Immunohistochemical studies demonstrated immunoreactivity for AFP and testosterone in Leydig's cells only. Secretion of AFP by Sertoli-Leydig cell tumors has rarely been mentioned previously, and its mechanism is difficult to explain. Non-germ cell tumors must be considered in the differential diagnosis of ovarian tumors with elevated serum AFP levels.     

Patterns of exochorion ornaments on eggs of seven South American species of Lutzomyia sand flies (Diptera: Psychodidae)

The patterns of exochorion ornaments on eggs of seven South American Lutzomyia sand fly species were analyzed by scanning electron microscopy (SEM): Lutzomyia (Lutzomyia) cruzi (Mangabeira 1938), Lutzomyia (Micropygomyia) evandroi (Costa Lima and Antunes 1936), L. (Nyssomyia) intermedia (Lutz and Neiva 1912), L. longipalpis (Lutz and Neiva 1912), L. migonei (Franca 1920), L. (Nyssomyia) neivai (Pinto 1926), and L. renei (Martins, Falcao, and Silva 1957). Different patterns were observed, which showed the distinction between some species. Egg ornaments in L. cruzi and L. longipalpis appear as single, parallel, unconnected ridges, whereas eggs of L. migonei appear as single, parallel, connected ridges. Eggs of L. (Nyssomyia) intermedia and L. (N.) neivai present a new variation of the single, unconnected, parallel ridges pattern: small tubercles are present, distributed between the ridges. Eggs of L. renei present an elliptical pattern, with most structures connected by straight ridges. Eggs of L. (M.) evandroi present a polygonal pattern, with alternate rows of small and large hexagons. Our data emphasize the advantages of the SEM approach in the study of the exochorion patterns of Lutzomyia eggs and in the distinction of the sand fly species.     

Immune response in human visceral leishmaniasis: analysis of the correlation between innate immunity cytokine profile and disease outcome

We investigated the cytokine profile of cells of the innate immune response and its association with active (ACT), asymptomatic (AS) and cured (CUR) human visceral leishmaniasis (VL), as well as noninfected (NI) subjects. The frequency of cytokine-producing cells was determined after short-term in vitro incubation of whole peripheral blood samples with soluble Leishmania antigen (SLA). Our data demonstrated a predominant type 2 cytokine profile in NI and ACT. In NI, we observed an increase of IL-4+ neutrophils, IL-10+ eosinophils besides a decrease of tumour necrosis factor (TNF)-alpha+ eosinophils/monocytes. Yet in ACT, we observed an increase of IL-4+ neutrophils and natural killer (NK) cells and IL-10+ monocytes, a reduced frequency of IL-12+ and IFN-gamma+ eosinophils and lower levels of TNF-alpha+ and IL-12+ monocytes. AS presented a mixed profile, characterized by an increase of IFN-gamma+ neutrophils/eosinophils and NK cells, of IL-12+ eosinophils/monocytes, as well as increase of IL-4+ neutrophils and NK cells and IL-10+ eosinophils/monocytes. In contrast, CUR was characterized by a type 1 response with an increase of IFN-gamma+ neutrophils/eosinophils and NK cells, associated with an increase in IL-12+ monocytes. In conclusion, we show a correlation between innate immune cytokine patterns and clinical status of VL, suggesting that these cells, in addition to other factors, may contribute to the cytokine microenvironment in which Leishmania-specific T cells are primed and to disease outcome.