Mammalian nuclear transplantation to Germinal Vesicle stage Xenopus oocytes - a method for quantitative transcriptional reprogramming.

Full-grown Xenopus oocytes in first meiotic prophase contain an immensely enlarged nucleus, the Germinal Vesicle (GV), that can be injected with several hundred somatic cell nuclei. When the nuclei of mammalian somatic cells or cultured cell lines are injected into a GV, a wide range of genes that are not transcribed in the donor cells, including pluripotency genes, start to be transcriptionally activated, and synthesize primary transcripts continuously for several days. Because of the large size and abundance of Xenopus laevis oocytes, this experimental system offers an opportunity to understand the mechanisms by which somatic cell nuclei can be reprogrammed to transcribe genes characteristic of oocytes and early embryos. The use of mammalian nuclei ensures that there is no background of endogenous maternal transcripts of the kind that are induced. The induced gene transcription takes place in the absence of cell division or DNA synthesis and does not require protein synthesis. Here we summarize new as well as established results that characterize this experimental system. In particular, we describe optimal conditions for transplanting somatic nuclei to oocytes and for the efficient activation of transcription by transplanted nuclei. We make a quantitative determination of transcript numbers for pluripotency and housekeeping genes, comparing cultured somatic cell nuclei with those of embryonic stem cells. Surprisingly we find that the transcriptional activation of somatic nuclei differs substantially from one donor cell-type to another and in respect of different pluripotency genes. We also determine the efficiency of an injected mRNA translation into protein.

Elements in rice on the Swedish market: part 2. Chromium, copper, iron, manganese, platinum, rubidium, selenium and zinc.

A survey of the levels of some essential and non-essential trace elements in different types of rice available on the Swedish retail market was carried out in 2001-03. The types of rice included long and short grain, brown, white, and parboiled white. The mean levels found were: chromium (Cr) = 0.008 mg kg(-1), copper (Cu) = 1.9 mg kg(-1), iron (Fe) = 4.7 mg kg(-1), manganese (Mn) = 16 mg kg(-1), platinum (Pt) < 0.0003 mg kg(-1), rubidium (Rb) = 3.3 mg kg(-1), selenium (Se) =0.1 mg kg(-1); and zinc (Zn) = 15 mg kg(-1). Inductively coupled plasma-mass spectrometry (ICP-MS) was used for the determination of Pt, Rb, and Se, after acid digestion. All other elements were determined using atomic absorption spectrometry (AAS) after dry ashing. Intake calculations were performed and it was concluded that rice may contribute considerably to the daily requirements of the essential elements Cu, Fe, Mn, Se, and Zn if rice consumption is high. The levels of some elements, e.g. Fe and Mn, were significantly higher in brown compared with white rice.

Elements in rice from the Swedish market: 1. Cadmium, lead and arsenic (total and inorganic).

A survey of the levels of cadmium, lead and arsenic in different types of rice available on the Swedish retail market was carried out in 2001--03. The types of rice included long and short grain, brown, white, and parboiled white rice. The mean levels found were as follows: total As: 0.20 mg kg(-1), inorganic As: 0.11 mg kg(-1); Cd: 0.024 mg kg(-1); and Pb: 0.004 mg kg(-1). ICP-MS was used for the determination of As (total and inorganic) after acid digestion. Lead and cadmium were determined using graphite furnace atomic absorption spectrometry (GFAAS) after dry ashing. In countries where rice is a staple food, it may represent a significant contribution in relation to the provisional tolerable weekly intake for Cd and inorganic As.

The levels of zinc, copper, manganese, selenium, chromium, nickel, cobalt, and aluminium in the meat, liver and kidney of swedish pigs and cattle.

The levels of zinc, copper, manganese, selenium, chromium, nickel, cobalt, and aluminium were determined in samples of meat, liver and kidney from pigs and cattle from Swedish slaughterhouses. The results have been compared with those reported from other investigations carried out in recent years. For zinc, copper, manganese and selenium, the mean levels found in the different tissues were very similar to those reported in previous studies. The mean levels of chromium and nickel were found to be in the range less than 0.010-0.015 mg/kg, regardless of the type of tissue. Cobalt levels ranged from 0.001 to 0.043 mg/kg, the highest levels being in the liver samples. The mean levels of aluminium were between 0.024 and 0.068 mg/kg. Tissues from cattle contained approximately twice as much aluminium as tissues from pigs. Analytical quality assurance was carried out by analysing standard reference materials. A great effort was made to minimize and check contamination. This may partly explain the fact that the levels of chromium, nickel, and aluminium found in the present study are among the lowest reported.