Pitfalls in molecular diagnosis in a family with severe factor VII (FVII) deficiency—misdiagnosis by direct sequence analysis using a PCR product

Molecular diagnostic tests are becoming a routine analysis in many laboratories. These modern analyses are widely used in clinical medicine, forensic, genetic and prenatal diagnosis and also in preimplantation genetic diagnosis. The accuracy of analysis is highly dependent on the success achieved in minimising genotyping errors. The pitfalls in molecular diagnostic tests can be due to a simple technique such as the polymerase chain reaction (PCR) used universally. This technique is routinely used for its apparent accuracy, but it is also a well-known source of errors. We report an error introduced during PCR reaction that leads to a wrong sequence result and consequently to a ‘false’ molecular result in a next prenatal diagnosis in a family with severe factor VII (FVII) deficiency. This error was verified using an unsuitable primer design in a rich repetitive sequence of the FVII gene that leads to a false annealing and then to a wrong molecular diagnosis. It is essential to link closely molecular data with clinical and phenotype analysis in order to avoid false-negative or false-positive results, which is of great importance to diagnosis and molecular prevention. Copyright © 2003 John Wiley & Sons, Ltd.     

Melatonin ameliorates chronic copper-induced lung injury

Environmental Science and Pollution Research - Copper (Cu) is an important trace element required for several biological processes. The use of copper is increasing gradually in several...     

On the crucial importance of a small bird: The ecosystem services of the little auk (<Emphasis Type="Italic">Alle alle</Emphasis>) population in Northwest Greenland in a long-term perspective

The little auk is the most numerous seabird in the North Atlantic and its most important breeding area is the eastern shores of the North Water polynya. Here, a population of an estimated 33 million pairs breeds in huge colonies and significantly shapes the ecosystem. Archaeological remains in the colonies document that the little auk has been harvested over millennia. Anthropological research discloses how the little auk has a role both as social engineer and as a significant resource for the Inughuit today. The hunting can be practiced without costly equipment, and has no gender and age discrimination in contrast to the dominant hunt for marine mammals. Little auks are ecological engineers in the sense that they transport vast amounts of nutrients from sea to land, where the nutrients are deposited as guano. Here, the fertilized vegetation provides important foraging opportunities for hares, geese, fox, reindeer, and the introduced muskox. We estimate that the relative muskox density is ten times higher within 1 km of little auk fertilized vegetation hotspots.     

Copper distribution in surface and subsurface soil horizons

The horizons of four natural soils were treated with Cu²⁺ in an acid medium to study the retention capacity of Cu. The possible mineralogical changes arising because of the treatment were also studied. The soil properties and characteristics with the greatest influence on the metal retention and its distribution among the different soil fractions were determined. Crystalline phases of each horizon were determined by X-ray diffraction (XDR). The morphology, structural distribution and particle chemical composition of soil samples were investigated using field emission scanning electron microscopy. Cu distribution in the different geochemical phases of the soil was studied using a sequential extraction. The treatment led to an increase in the amorphous phases and the formation of new crystalline phases, such as rouaite (Cu₂(NO₃)(OH)₃) and nitratine (NaNO₃). Cu was also found superficially sorbed on amorphous hydroxy compounds of Fe that interact with albite, muscovite and gibbsite, and also on spherical and curved particles of aluminium clays. The largest amount of Cu retained was in an exchangeable form, and the smallest amount associated with the crystalline Fe oxides and residual fraction. In the surface horizons, the predominant Cu retention process is complexation in organomineral associations, while in the subsurface horizons it is adsorption.     

Comparative hepatic and renal toxicity of cadmium in male and female rats

Rats (male and female) were exposed to 0.5 mg/kg and 1 mg/kg cadmium as cadmium chloride for 3 days and subsequently sacrificed for cadmium concentration and other biochemical variables indicative of hepatic and renal damage. The absorption of cadmium was supported by biochemical changes, which were significantly higher in females than in males. This could be due to higher rate of intestinal absorption of cadmium in females than males. Male and female rats both showed relatively higher cadmium concentration in kidneys than in liver. Female rats also showed the similar trend in tissue metal levels as compared to male rats. However, hepatic and renal histopathological observations showed that female rats suffered from severe hepatic injury like hydropic degeneration of hepatocytes, granulation, bile duct proliferation etc. In comparison to female rats, male rats did not show much remarkable changes. Renal damage was more prominent in female than male in the form of renal tubular damage; most of the tubular nuclei were pyknotic, congestion of the boundary of cortex and medulla etc. The results suggested that females were comparatively more vulnerable to the toxic effects of cadmium than males.     

Uptake and release of cesium-137 by five plant species as influenced by soil amendments in field experiments

Phytoextraction field experiments were conducted on soil contaminated with 0.39 to 8.7 Bq/g of 137Cs to determine the capacity of five plant species to accumulate 137Cs and the effects of three soil treatments on uptake. The plants tested were redroot pigweed (Amaranthus retroflexus L. var. aureus); a mixture of redroot pigweed and spreading pigweed (A. graecizans L.); purple amaranth (A. cruteus L.) x Powell's amaranth (A. powellii S. Watson), referred to here as the amaranth hybrid; Indian mustard [Brassica juncea (L.) Czern.]; and cabbage (Brassica oleracea L. var. capitata). For control plants, the concentration ratios (CR) of 137Cs were greatest for redroot pigweed and the amaranth hybrid, with average CR values of 1.0 +/- 0.24 and 0.95 +/- 0.14, respectively. The lowest value was for Indian mustard at 0.36 +/- 0.10. The soil treatments included (i) application of NH4NO3 solution to the soil after plants had matured, (ii) addition of composted manure to increase organic matter content of the soil, (iii) combination of the manure and ammonium solution treatments, and (iv) controls. The ammonium solution gave little overall increase in accumulation of 137Cs. The use of composted manure also had little influence, but the combination of the composted manure with application of ammonium solutions had a distinctly negative effect on plant uptake of 137Cs. On average the fraction of 137Cs taken up from the soil was reduced by 57.4 +/- 1.2% compared with controls. This was the result of release of competing ions, primarily Ca, from the manure and was observed across all five plant species tested. The application of ammonium solution took place in the last two weeks before harvest. The reduction of plant 137Cs content, by addition of the ammonium solution, as it interacted with the manure, indicates that substantial quantities 137Cs can be released from the shoots of plants as a result of sudden changes in soil solution chemistry.     

Zearalenone screening of human breast milk from the Naples area

The mycotoxin zearalenone exhibits estrogenic and anabolic properties in several animal species, humans included. Food contamination by zearalenone is caused either by direct contamination of grains, fruits, and their based-products or by “carry-over” of mycotoxins in animal tissues, milk, and eggs after intake of contaminated feedstuff. Now, a survey on zearalenone contamination in breast milk of healthy primiparous women living in the Naples countryside was conducted. From 47 healthy primiparous women, breast milk samples were collected within the first six weeks after delivery as well as clinical data of mother–newborn pairs. Breast milk analyses were performed with both competitive indirect enzyme-linked immunosorbent assay and high-performance liquid chromatography with fluorescence detection. At 36.9 ± 2.6 days after vaginal delivery, the mean zearalenone contents of the breast milk samples were 1.13 ± 0.34 µg/L. The zearalenone levels correlated with both mother weights before pregnancy (r = ?0.506; P < 0.001) and at delivery (r = ?0.351; P < 0.05). The present results indicate that breast milk may be contaminated with zearalenone.