Cryptosporidiosis in the V Region Chile. III. Study of malnourished patients, 1985-1987

Frequency of infection by Cryptosporidium sp. in 1,039 faecal smears stained by Ziehl-Neelsen, obtained from undernourished patients of a Nutritional Recovery Center and an ambulatory undernourished center from the Fifth Region, Chile, were studied. All underwent a coproparasitological examination by the modified Telemann method. Thirty eight (3.7%) patients infected by the parasite were detected, with an overall frequency of 8.5% among patients of the Nutritional Recovery Center and 1.9% among the ambulatory patients; this difference was statistically significant. The highest percentage of positive results were detected among the younger milk feeding infants. Also the percentage of difference among these and the older milk feeding infants (3.7%) was statistically significant. Association of Cryptosporidium sp. and Giardia lamblia was observed in 6 ambulatory patients (2.3%).     

3D structure of bovine pancreatic ribonuclease A in aqueous solution: An approach to tertiary structure determination from a small basis of1H NMR NOE correlations

Summary A method is proposed to generate initial structures in cases where the distance geometry method may fail, such as when the set of¹H NMR NOE-based distance constraints is small in relation to the size of the protein. The method introduces an initial correlation between the and backbone angles (based on empirical observations) which is relaxed in later stages of the calculation. The obtained initial structures are refined by well-established methods of energy minimization and restrained molecular dynamics. The method is applied to determine the solution structure of Ribonuclease A (124 residues) from a NOE basis consisting of 467 NOE cross-correlations (97 intra-residue, 206 sequential, 23 medium-range and 141 long-range) obtained at 360 MHz. The global shape and backbone overall fold of the eight final refined structures are close to those shown by the crystal structure. A meaningful difference in the positioning of the catalytically important His¹¹⁹ side chain in the solution and crystal structures has been detected.     

The ichthyoplankton of a seasonally closed estuary in temperate Australia. Does an extended period of opening influence species composition?

Fish eggs and larvae were collected monthly between September 1987 and April 1989 from sites throughout the main basin and within the saline regions of the two main tributary rivers of Wilson Inlet, a seasonally closed estuary in south-western Australia. Of the eggs, 43.7% belonged to Engraulis australis (Shaw) and the rest to unidentified teleosts. The larval fish assemblage comprised 17 families represented by 25 species. The Gobiidae contained the highest number of species (four) and contributed approximately 57% of all larvae caught. Pseudogobius olorum (Sauvage) and E. australis were the most abundant species, contributing 43.9 and 27.9% to the total larval catch, respectively. The larvae of species which breed within Wilson Inlet dominated the assemblage, both in terms of number of species (64%) and contribution to total catch (99.9%). The numbers of the eight marine species and one freshwater species represented in the ichthyoplankton were very low. Classification and multi-dimensional scaling ordination showed that the composition of the larval fish fauna at the various sites during a period when the estuary remained open to the sea (December 1988-April 1989) was similar to that of the corresponding sites during the same period in the previous year when the estuary had become closed (December 1987-April 1988). This can be attributed to the spatial distribution, time of occurrence and abundance of estuarine-spawned larvae being similar in the two periods and to the rarity of marine-spawned larvae, even in the spring and summer of 1988/1989 when the estuary was open for the whole time when most marine teleosts spawn in south-western Australia. The low occurrence of marine-spawned larvae in Wilson Inlet reflects the fact that tidal water movement within the basin of the system is so small that it is unable to facilitate the transport and dispersion of larvae. The ichthyoplankton of Wilson Inlet resembles that of other poorly-flushed estuaries in that it is low in species richness and dominated by estuarine-spawned larvae.     

Reproductive biology and larval morphology of the marine plotosid Cnidoglanis macrocephalus (Teleostei) in a seasonally closed Australian estuary

Monthly trends shown by gonadosomatic indices, the prevalence of the different gonadal stages, and the size distribution of the oocytes, indicate that the large marine and commercially important plotosid Cnidoglanis macrocephalus spawns in Wilson Inlet between October and January. The conclusion that spawning occurs within this seasonally closed estuary was confirmed by the presence of males in large nests and by the capture of newly-hatched, yolk sac larvae from one of those nests. The fact that C. macrocephalus, which is also widely distributed in coastal marine waters throughout much of southern Australia, can spawn within Wilson Inlet would be of particular value to this species in those periods when closure of the estuary would preclude a seawards spawning migration. Sexual maturity is size dependent, with spawning rarely occurring before fish have reached a total length of 425 mm. Sexual maturity was attained by a few fish at the end of their second year, by several at the end of their third year and by most, if not all fish, at the end of their fourth year. Comparisons with data for the more northern and permanently open Swan Estuary indicate that C. macrocephalus also spawns within that system and that the spawning time of this species is related to water temperature. The adult male guards the larvae under its pelvic fins in burrows. The larvae increased in total length from 29 mm just after hatching to 43 mm in the 17–18 days after capture, during which time their yolk sac was resorbed. Details are given of the morphology, morphometrics, meristics and pigmentation of larval C. macrocephalus. In comparison with the larvae of three other plotosid genera, the larva of C. macrocephalus is far larger in size and more developed at hatching and takes a shorter time to transform into a juvenile.     

Optimizing Human Synovial Fluid Preparation for Two-Dimensional Gel Electrophoresis

Background

Prenatal screening for Down Syndrome (DS) would benefit from an increased number of biomarkers to improve sensitivity and specificity. Improving sensitivity and specificity would decrease the need for potentially risky invasive diagnostic procedures.

Results

We have performed an in depth two-dimensional difference gel electrophoresis (2D DIGE) study to identify potential biomarkers. We have used maternal plasma samples obtained from first and second trimesters from mothers carrying DS affected fetuses compared with mothers carrying normal fetuses. Plasma samples were albumin/IgG depleted and expanded pH ranges of pH 4.5 - 5.5, pH 5.3 - 6.5 and pH 6 - 9 were used for two-dimensional gel electrophoresis (2DE). We found no differentially expressed proteins in the first trimester between the two groups. Significant up-regulation of ceruloplasmin, inter-alpha-trypsin inhibitor heavy chain H4, complement proteins C1s subcomponent, C4-A, C5, and C9 and kininogen 1 were detected in the second trimester in maternal plasma samples where a DS affected fetus was being carried. However, ceruloplasmin could not be confirmed as being consistently up-regulated in DS affected pregnancies by Western blotting.

Conclusions

Despite the in depth 2DE approach used in this study the results underline the deficiencies of gel-based proteomics for detection of plasma biomarkers. Gel-free approaches may be more productive to increase the number of plasma biomarkers for DS for non-invasive prenatal screening and diagnosis.     

Plasminogen activator and collagenase production by cultured capillary endothelial cells

Cultured bovine capillary endothelial (BCE) cells produce low levels of collagenolytic activity and significant amounts of the serine protease plasminogen activator (PA). When grown in the presence of nanomolar quantities of the tumor promoter 12-O-tetradecanoyl phorbol-13-acetate (TPA), BCE cells produced 5-15 times more collagenolytic activity and 2-10 times more PA than untreated cells. The enhanced production of these enzymes was dependent on the dose of TPA used, with maximal response at 10(-7) to 10(-8) M. Phorbol didecanoate (PDD), an analog of TPA which is an active tumor promoter, also increased protease production. 4-O-methyl-TPA and 4α-PDD, two analogs of TPA which are inactive as tumor promoters, had no effect on protease production. Increased PA and collagenase activities were detected within 7.5 and 19 h, respectively, after the addition of TPA. The TPA-stimulated BCE cells synthesized a urokinase-type PA and a typical vertebrate collagenase. BCE cells were compared with bovine aortic endothelial (BAE) cells and bovine embryonic skin (BES) fibroblasts with respect to their production of protease in response to TPA. Under normal growth conditions, low levels of collagenolyic activity were detected in the culture fluids from BCE, BAE, and BES cells. BCE cells produced 5-13 times the basal levels of collagenolytic activity in response to TPA, whereas BAE cells and BES fibroblasts showed a minimal response to TPA. Both BCE and BAE cells exhibited relatively high basal levels of PA, the production of which was stimulated approximately threefold by the addition of TPA. The observation that BCE cells and not BAE cells produced high levels of both PA and collagenase activities in response to TPA demonstrates a significant difference between these two types of endothelial cells and suggests that the enhanced detectable activities are a property unique to bovine capillary and microvessel and endothelial cells.