Glomerular anionic charge in congenital nephrotic syndrome of the Finnish type

Summary Decrease of the anionic charge of the glomerular basement membrane and especially the reduced amount of heparan sulphate proteoglycan in the lamina rara externa has been suggested to be the basic pathogenetic defect in congenital nephrotic syndrome. In the present study the anionic charge of glomeruli was examined in the congenital nephrotic syndrome of the Finnish type and in controls using cationic stains (polyethyleneimine, Ruthenium Red) in electron microscopy. Chondroitinase and heparinase treatments were used to characterize further the anionic elements detected. Scanning electron microscopy (SEM) was used in addition to transmission electron microscopy (TEM) to examine the tridimensional structure and secondary changes of podocytes in this syndrome. The number (mean ± SD) of polyethyleneimine granules per 1 μm length of lamina rara externa of the glomerular basement membrane was 24.9 ± 4.5 in control and 2.32 ± 4.3 in congenital nephrotic syndrome subjects. The Ruthenium Red staining pattern was closely similar in syndrome and control kidneys. The granules evident after staining with either cationic stain were seen after chondroitinase but not after heparinase treatment in control as well as in syndrome patient kidney samples. No denuded areas of basement membrane in 42 glomeruli from four syndrome patients were found in SEM. In conclusion, the amount of anionic sites in the lamina rara externa as detected by either cationic stain was comparable to controls. These results do not support the hypothesis of decreased anionic sites in the lamina rara externa of the glomerular basement membrane in congenital nephrotic syndrome of the Finnish type.     

The role of bacteria in the nutrient exchange between sediment and water in a flow-through system

The contribution of bacteria to phosphorus (P) and nitrogen (N ) release from, or retention in, sediment was studied in a flow-through system. Live and formaldehyde-killed sediment communities were incubated in 25-liter bottles with a continuous flow of P- or P + N-enriched water. Sediment bacteria in the killed communities were inhibited by adding formaldehyde (final concentration 0.04% v/v) to the sediment before the start of the experiment. Bacterial activity in the live sediments measured with [³H]thymidine and [¹⁴C]leucine incorporation techniques did not change essentially during the experiment period (7–8 days). Chemical mechanisms were found to be of principal importance in PO₄-P retention in the sediment. In the live samples, the net retention of PO₄-P was lower than in the killed samples, which was likely due to the reduced O₂ conditions in the sediment as a consequence of bacterial mineralization. In total P exchange, however, bacteria increased the retention rate by recycling dissolved organic P in the sediment. In the live communities the retention of N was very efficient, and all the introduced NH₄ -N and NO₃-N was immobilized by sediment bacteria. Nitrogen enrichment, however, did not alter the P exchange rates. The gradual emergence of bacterial activity (and grazing) in the killed communities, subsequent to the dilution of formaldehyde concentration, enhanced the release of PO₄-P and NH₄-N from sediment.     

Pharmacological activity of the Samoan ethnopharmacopoeia

The Samoan ethnopharmacopoeia was surveyed for pharmacological activity using broad in vitro and in vivo screens. Residues of 74 different plant species were tested for activity in a Hippocratic screen and in a guinea pig ileum test. Over 86% of the plant species exhibited pharmacological activity. This high percentage of active species strongly supports the belief that ethnobotanical analyses of indigenous floras are more likely than random screens to efficiently identify plants likely to yield new drugs. However, collaboration of ethnobotanists and pharmacognosists is necessary to adequately move promising plants from the realm of indigenous knowledge systems to the laboratories of Western pharmacology.     

The immunological status of breast cancer patients during treatment with a new antiestrogen,toremifene

Summary The immune status of breast cancer patients was followed during antiestrogen treatment for at least 1 year or until progression of the disease. Twelve postmenopausal women with advanced estrogen-receptor-positive breast cancer were treated with a novel antiestrogen, toremifene. Immune functions were determined before the start of the treatment and at 3, 6, and 12 months. For NK cell cytotoxicity testing there were 74 healthy controls and for T cell subset measurements 28 healthy controls. No statistically significant changes in the T cell subsets or NK cell cytotoxicity were observed during treatment. However, throughout toremifene treatment patients had fewer CD4 cells (T helper lymphocytes) than did the controls. Cancer patients had higher pretreatment B cell values than the controls,P = 0.01, but during the first months of toremifene treatment B cell values decreased and remained within the normal range thereafter. A positive effect on mitogen-stimulation tests with phytohemagglutinin (PHA) and concanavalin A (ConA) was observed during the first months of treatment (P = 0.01 for PHA and 0.03 for log [ConA] and a stabilization at the higher level thereafter. These results indicate that toremifene has a stimulatory effect on cell-mediated immunity in breast cancer patients.     

Disruption of certain aleurone grains by various homogenization agents

Although aleurone grains of certain oilseeds (e. g. peanuts)are exceedingly sensitive to aqueous solutions, it has beenreported that aleurone grains maintain their integrity whenhomogenized in an aqueous solution containing 15% Carbowax 6000(PEG-6000). Presumably this substance acts as an osmotic agent.We have been unable to confirm this result using microscopicand chemical studies. However, we found that PEG-6000 acts asa protein precipitant; therefore, it is suggested that the pelletobtained upon centrifugation of the homogenate is mostly proteinprecipitate rather than intact aleurone grains. ¹Fulbright-Hayes Research Scholar 1972–1973. Present address:Dept. of Cell Biology, Univ. of Jyvaskyla, Vapaudenkatu 4, SF-40100,Jyvaskyla 10, Finland. (Received May 13, 1974; )     

Preferential synthesis of heptaacyl lipopolysaccharide by the ssc permeability mutant of Salmonella typhimurium.

In Salmonella typhimurium, a chromosomal gene termed ssc has been shown to cause an antibiotic-supersensitive phenotype. We studied the effect of the ssc gene on the chemical composition of the lipopolysaccharide component, using a thermosensitive ssc1 mutant (SH7622) that grows poorly at 42 degrees C. Analysis of the lipopolysaccharide by various techniques including fast-atom-bombardment mass spectrometry of lipid A, and determination of the type of linkage of fatty acids, revealed a profound temperature-dependent effect associated with the ssc1 mutation. At the non-permissive temperature, SH7622 contained hexadecanoic acid in the majority of lipid A molecules, resulting in the exclusive presence of heptaacyl lipopolysaccharide. This effect was largely reversed by the introduction of the cloned wild-type ssc gene to SH7622 and much reduced by growth of SH7622 at 37 degrees C.     

The lowering of hepatic fatty acid uptake improves liver function and insulin sensitivity without affecting hepatic fat content in humans

Lipolysis may regulate liver free fatty acid (FFA) uptake and triglyceride accumulation; both are potential causes of insulin resistance and liver damage. We evaluated whether 1) systemic FFA release is the major determinant of liver FFA uptake in fasting humans in vivo and 2) the beneficial metabolic effects of FFA lowering can be explained by a reduction in liver triglyceride content. Sixteen healthy subjects were subdivided in two groups of similar characteristics to undergo positron emission tomography with [(11)C]acetate and [(11)C]palmitate to quantify liver FFA metabolism (n = 8), or magnetic resonance spectroscopy (MRS) to measure hepatic fat content (n = 8), before and after the acute lowering of circulating FFAs by using the antilipolytic agent acipimox. MRS was again repeated after a 1-wk treatment period. Acipimox suppressed FFA levels while stimulating hepatic fractional extraction of FFAs (P < 0.05). As a result, fasting liver FFA uptake was decreased by 79% (P = 0.0002) in tight association with lipolysis (r = 0.996, P < 0.0001). The 1-wk treatment induced a significant improvement in systemic (+30%) and liver (+70%) insulin sensitivity (P < 0.05) and decreased circulating triglycerides (-20%, P = 0.06) and liver enzymes (ALT -20%, P = 0.03). No change in liver fat content was observed after either acute or sustained FFA suppression. We conclude that acute and sustained inhibitions of lipolysis and liver FFA uptake fail to deplete liver fat in healthy human subjects. Liver FFA uptake was decreased in proportion to FFA delivery. As a consequence, liver and systemic insulin sensitivity were improved, together with liver function, independently of changes in hepatic triglyceride accumulation.     

Gap-Directed Translesion DNA Synthesis of an Abasic Site on Circular DNA Templates by a Human Replication Complex

DNA polymerase ε (pol ε) is believed to be the leading strand replicase in eukaryotes whereas pols λ and β are thought to be mainly involved in re-synthesis steps of DNA repair. DNA elongation by the human pol ε is halted by an abasic site (apurinic/apyrimidinic (AP) site). We have previously reported that human pols λ, β and η can perform translesion synthesis (TLS) of an AP site in the presence of pol ε. In the case of pol λ and β, this TLS requires the presence of a gap downstream from the product synthetized by the ε replicase. However, since these studies were conducted exclusively with a linear DNA template, we decided to test whether the structure of the template could influence the capacity of the pols ε, λ, β and η to perform TLS of an AP site. Therefore, we have investigated the replication of damaged “minicircle” DNA templates. In addition, replication of circular DNA requires, beyond DNA pols, the processivity clamp PCNA, the clamp loader replication factor C (RFC), and the accessory proteins replication protein A (RPA). Finally we have compared the capacity of unmodified versus monoubiquitinated PCNA in sustaining TLS by pols λ and η on a circular template. Our results indicate that in vitro gap-directed TLS synthesis by pols λ and β in the presence of pol ε, RPA and PCNA is unaffected by the structure of the DNA template. Moreover, monoubiquitination of PCNA does not affect TLS by pol λ while it appears to slightly stimulate TLS by pol η.