Cystic fibrosis gene mutations and linked RFLPs in the Slovenian population.

The authors used polymerase chain reaction to analyse 56 Slovenian cystic fibrosis (CF) chromosomes for the presence of delta F508 and eight other most frequent mutations located in exons 7,11 and 20 (R347P, R334W, G551D, R553X, S549RA, S549RT, S549I and S1255X) of the CF gene. We also determined the frequency of haplotypes associated with CF for six linked RFLP markers (MetD/TaqI, MetH/TaqI, XV-2c/TaqI, KM-19/PstI, MP6d9/MspI and J3.11/MspI) in 27 Slovenian CF families. delta F508 mutation was present in 55.4 percent of the CF chromosomes. No case of the other mutations were detected in the sample of tested CF chromosomes. A very high degree of association (0.88) has been found between DNA marker MetH and CF (as measured by the Yule's association coefficient) in our population. Using the RFLP markers XV-2c and KM-19, we found that 85% of delta F508 mutated chromosomes have a single 1 2 (B) haplotype, and that this haplotype is present on only 15.4 percent of CF chromosomes without this deletion.     

Effects of various environmental conditions on growth and reproduction of the sea hareAplysia oculifera (Adams and Reeve, 1850)

We studied the influences of food type, food quantity, water currents, starvation and light on growth and reproduction of the sea hareaplysia oculifera (Adams and Reeve, 1850) under laboratory conditions. Out of five species of algae served as food,Enteromorpha intestinalis promoted the fastest growth ofA. oculifera, Ulva spp. slower growth,Cladophora sp. allowed maintenance spp. slower growth,Cladophora sp. allowed maintenance of steady body mass, and the brown algaeColpomenia sp. andPadina pavonia were rejected by the sea hares. When sea hares were exposed to four levels of water currents, growth rates decreased as water currents increased. Sea hares fed on 50% ration grew slower than those fed on 100% ration (ad libitum). During 10 days of starvation sea hares lost weight, but when subsequently fed 100% ration they recovered and grew at a rate similar to those fed continuously with 100% ration. Under shade and under natural sunlight sea hares grew at the same rates. Whenever growth rates decreased, sea hares began to spawn at a smaller body size.A. oculifera demonstrated physiological plasticity that adapted them to varied and unpredictable environmental conditions. At different conditions of food availability they applied different tactics of resource allocation between growth and reproduction.     

Isolation and sequence analysis of the genomic DNA fragment encoding an aspartic proteinase inhibitor homologue from potato (Solanum tuberosum L.)

A genomic DNA clone encoding an aspartic proteinase inhibitor of potato was isolated from a lambda EMBL3 phage library using the aspartic proteinase inhibitor cDNA as a hybridization probe. The gene has all characteristic sequences normally found in eucaryotic genes. Typical CAAT and TATA box sequences were found in the 5-upstream region. In this part are also two putative regulatory AGGA box sequences located. In the genomic sequence there are no intron sequences interrupting the coding region. An open reading frame of the gene encodes a precursor protein of 217 amino acids which shows high percent identity with the aspartic proteinase inhibitor cDNA.     

Multi-proxy analysis of waterlogged preserved Late Neolithic canine excrements

Vegetation History and Archaeobotany - Multi-proxy analysis of the coprolites which were found during excavations at two Late Neolithic (fourth millennium bc) pile-dwelling sites (Črnelnik and...     

Secreted Protein Acidic and Rich in Cysteine (SPARC) Exacerbates Colonic Inflammatory Symptoms in Dextran Sodium Sulphate-Induced Murine Colitis

Background

Secreted Protein Acidic and Rich in Cysteine (SPARC) is expressed during tissue repair and regulates cellular proliferation, migration and cytokine expression. The aim was to determine if SPARC modifies intestinal inflammation.

Methods

Wild-type (WT) and SPARC-null (KO) mice received 3% dextran sodium sulphate (DSS) for 7 days. Inflammation was assessed endoscopically, clinically and histologically. IL-1β, IL-4, IL-5, IL-6, IL-10, IL-13, IL-17A, IL-12/IL23p40, TNF-α, IFN-γ, RANTES, MCP-1, MIP-1α, MIP-1β, MIG and TGF-β1 levels were measured by ELISA and cytometric bead array. Inflammatory cells were characterised by CD68, Ly6G, F4/80 and CD11b immunofluorescence staining and regulatory T cells from spleen and mesenteric lymph nodes were assessed by flow cytometry.

Results

KO mice had less weight loss and diarrhoea with less endoscopic and histological inflammation than WT animals. By day 35, all (n = 13) KO animals completely resolved the inflammation compared to 7 of 14 WT mice (p<0.01). Compared to WTs, KO animals at day 7 had less IL1β (p = 0.025) and MIG (p = 0.031) with higher TGFβ1 (p = 0.017) expression and a greater percentage of FoxP3+ regulatory T cells in the spleen and draining lymph nodes of KO animals (p<0.01). KO mice also had fewer CD68+ and F4/80+ macrophages, Ly6G+ neutrophils and CD11b+ cells infiltrating the inflamed colon.

Conclusions

Compared to WT, SPARC KO mice had less inflammation with fewer inflammatory cells and more regulatory T cells. Together, with increased TGF-β1 levels, this could aid in the more rapid resolution of inflammation and restoration of the intestinal mucosa suggesting that the presence of SPARC increases intestinal inflammation.     

Actual temperature during and thermal response after whole-body cryotherapy in cryo-cabin

Whole-body cryotherapy (WBC) involves exposing minimally dressed participants to very cold air (injecting liquid nitrogen with temperature −195 °C), either in a specially designed chamber (cryo-chamber) or cabin (cryo-cabin), for a short period of time. The aim of this study was to examine the actual temperature of the air in the cryo-cabin at different locations throughout the cabin by using human subjects and a manikin. Additionally, we monitored skin temperature before and for 60 min after the cryo-cabin session. Twelve subjects completed one 3 min cryo-cabin session. Temperature next to the skin was assessed during the session, while the skin temperature was monitored before, 3 min after and every 10 min for 60 min after completing the session. There was a statistically significant interaction (time×position) for temperature among the different body parts during the WBC, and for skin temperature among different body parts after the cryo-cabin session. Statistically significant time effects during and following cryo-cabin session were present for all body parts. We showed that actual temperature in the cryo-cabin is substantially different from the one reported by the manufacturer. Thermal response after cryo-cabin session is similar to response observed after cryo-chamber cold exposure reported in previously published studies. This could be of great practical value as cryo-cabins are less expensive and easier to use compared to cryo-chambers.