Are nanometric films of liquid undercooled interfacial water bio-relevant?

It is known that life processes below the melting point temperature can actively evolve and establish in micrometer-sized (and larger) veins and structures in ice and permafrost soil, filled with unfrozen water. Thermodynamic arguments and experimental results indicate the existence of much smaller nanometer sized thin films of undercooled liquid interfacial (ULI) water on surfaces of micrometer sized and larger mineral particles and microbes in icy environments far below the melting point temperature. This liquid interfacial water can be described in terms of a freezing point depression, which is due to the interfacial pressure of van der Waals forces. The physics behind the possibly also life supporting capability of nanometric films of undercooled liquid interfacial water, which also can "mantle" the surfaces of the much larger and micrometer-sized microbes, is discussed. As described, biological processes do not necessarily have to proceed in the "bulk"" of the thin interfacial water, as in "vinical" water and in the micrometer sized veins e.g., but they can be supported or are even made possible already by covering thin mantles of liquid interfacial water. These can provide liquid water for metabolic processes and act as carrier for the necessary transport of nutrients and waste. ULI water supports two different and possibly biologically relevant transport processes: 2D molecular diffusion in the interfacial film, and flow-like due to regelation. ULI-water, which is "lost" by transport into microbes, e.g., will be refilled from the neighbouring ice. In this way, the nanometric liquid environment of microbes in ULI-water is comparable to that of microbes in bulk water. Another probably also biologically relevant property of ULI is, depending on the hydrophobic or hydrophilic character of the surfaces, that it is of lower density (LDL) or higher density (HDL) than bulk water. Furthermore, capillary effects and ions in ULI-water solutions can support, enhance, and stabilize the formation of layers of interfacial water. A more detailed future investigation of the possible support of life processes by nanometric ULI water in ice is a challenge to current cryomicrobiology. Related results of Rivkina et al. [22] indeed indicate that life processes can remain active at water contents corresponding to about or less than two monolayers of ULI water.

Tissue-specific transcriptional activity of a pancreatic islet cell-specific enhancer sequence/Pax6-binding site determined in normal adult tissues in vivo using transgenic mice.

A pancreatic islet cell-specific enhancer sequence (PISCES) shared by the rat insulin-I, glucagon, and somatostatin genes binds the paired domain-containing transcription factor Pax6 and confers strong transcriptional activity in pancreatic islet cell lines. It was found recently that Pax6 plays a major role in islet development. In the present study, transgenic mice were used to investigate PISCES-mediated transcription in normal adult islets in vivo. In several independent mouse lines expressing a PISCES-luciferase reporter transgene, the PISCES motif directed gene expression in the adult eye, cerebellum, and discrete brain areas, consistent with the tissue distribution of Pax6. These tissues contain two Pax6 isoforms caused by alternative splicing, only one of which was found to bind the PISCES motif in electrophoretic mobility shift assays. No reporter gene expression was detected in adult pancreatic islets or in any other peripheral organ tested. RT-PCR analysis confirmed that Pax6 mRNA is present in adult islets. These results demonstrate that the PISCES motif is sufficient to direct highly tissue-specific gene expression in whole animals. The lack of PISCES-mediated transcription in adult islets indicates that the Pax6 protein(s) expressed in adult pancreatic islets function differently from the ones in the eye and cerebellum.

Transcriptional activity of domain A of the rat glucagon G3 element conferred by an islet-specific nuclear protein that also binds to similar pancreatic islet cell-specific enhancer sequences (PISCES).

A pancreatic islet cell-specific enhancer element in the rat glucagon gene, Glu-G3, contains two domains, one of which, domain A, has been shown to be necessary for Glu-G3 activity. In the present study, the functions of the isolated domain A of Glu-G3 were investigated by using transient reporter fusion gene expression and DNA binding assays. A single copy of domain A was transcriptionally inactive in glucagon-producing islet cell lines, whereas it did confer activity when combined with domain B, suggesting that Glu-G3 may be a bipartite element. Multiple copies of domain A did function independently as transcriptional enhancer in phenotypically distinct islet cell lines but not in several nonislet cell lines. Sequences (PISCES, pancreatic islet cell-specific enhancer sequences), similar to that of domain A of Glu-G3 and present in cell-specific control elements of the rat insulin I (Ins-E1) and rat somatostatin genes (SMS-UE), are shown to be required for transcriptional activity of these elements. In addition, a protein was detected in islet cell lines that bound to the PISCES motifs within Glu-G3, Ins-E1, and SMS-UE. These results support the view that cell-specific control elements of the glucagon, insulin, and somatostatin genes share a functional regulatory sequence, PISCES, and provide direct evidence for the existence of an islet-specific, PISCES-binding transcription factor or closely related proteins being involved in the coordinate expression of islet hormone genes.

Nuclear protein binding and functional activity of a variant insulin gene found in non-insulin-dependent diabetes mellitus.

In a subset of patients with non-insulin-dependent diabetes mellitus an 8-base pair (bp) repeat was found from -322 to -315 in the 5'-flanking region of the insulin gene. This 8-bp repeat is inserted into a sequence that is highly homologous to a sequence motif, called PISCES (pancreatic islet cell-specific enhancer sequences), found within cell-specific enhancer elements of the rat insulin I (Ins-E1, from -332 to -285), rat glucagon (Glu-G3) and rat somatostatin (SMS-UE) genes. The PISCES motif confers pancreatic islet-specific activity and is recognized by an islet-specific transcription factor (PISCES-BP). The consequences on functional activity and on protein binding of the 8-bp repeat sequence in the human insulin promoter was investigated. When fused to a reporter gene and transiently transfected into an insulin-producing islet cell line, the 8-bp repeat decreased basal transcriptional activity of the human insulin promoter (from -366 to +42) whereas the induction of promoter activity by cAMP was unaffected. The isolated rat Ins-E1 element was sufficient to confer basal transcriptional activity to a minimal promoter; the corresponding fragments of the normal and variant human insulin genes (from -329 to -288), however, were not. Using nuclear extracts in an electrophoretic mobility shift assay, it was found that PISCES-BP recognizes rat Ins-E1, but PISCES-BP binding to the corresponding normal and variant human insulin promoter fragments was not detectable and weak, respectively. However, a nuclear protein was found that binds to the variant but not normal human sequence. These data suggest that the 8-bp repeat in the variant human insulin promoter found in patients with non-insulin-dependent diabetes mellitus allows the binding of a nuclear protein that interferes with promoter function.

A formal model of diabetological terminology and its application for data entry.

This paper summarises the developmental activities for an electronic patient record system in diabetology based on GALEN technologies. It focusses on the modelling of primarily terminological medical knowledge of this subspecialty and describes its application for predictive data entry.

[Development and evaluation of a soccer focused complexity test]. / Entwicklung und Evaluierung eines fußballspezifischen Komplextests.

BACKGROUND: Soccer has increasingly become a technical - tactical (speed of activity!) and athletic sport over the last years. Simultaneously the new training and contest conditions result in new challenges to sports science and sports medicine evaluation. Complex, sports specific field tests for soccer exist rarely and only in low quality. OBJECTIVE: Development and evaluation of a standardised complexity test in soccer (FBKT) for assessment of the complex sports specific and league specific physical performance. MATERIALS AND METHODS: Two regional German soccer teams (Verbandsliga [VL], Oberliga [OL]) (n = 27) were assessed with FBKT. The assessment included lactate levels, heart rate as well as time and error frequency (penalty time per error: 5 s) for defined activity series at defined time points (heart rate: rest, after round 1, prior to round 2, E 0, E 2, E 6, E 10, E 14; lactate: rest, E 2, E 6, E 10, E 14). RESULTS: The heart rate (hr (max) = 200 min (-1)) and metabolic rate (lactate (max) = 17.1 mmol/l) were very excessively high and did only partially differ between the groups (heart rate difference at rest p = 0.005, prior to round 2: p = 0.014, E 6: p = 0.042). Furthermore no significant differences occurred in the following parameters: recovery potential (hr (E0 - E14): p = 0.560; lactate (Max-Min): p = 0.448), technical error (p = 0.384), sprint (p = 0.499), slalom dribbling (p = 0.310), time round 1 (p = 0.119), shots on goal (p = 0.585) and crosses (p = 0.676). Significant difference were only found in the parameters time round 2 (p = 0.004), time round 1 and round 2 (p = 0.013), overall time (running)(time + penalty)(time) (p = 0.022) and speed dribbling (p = 0.005). CONCLUSIONS: Soccer specific complex loads generate very high physical demamds. Although complex, the FBKT proved to be a practical assessment tool with high standardising potential. The FBKT gives plentiful information under competition - like conditions such as technical skills, speed, speed endurance or anaerobe capacity.

Interaction of the transcription factor CREB with pancreatic islet cell-specific enhancer elements.

The cAMP-responsive element (CRE)-binding transcription factor CREB confers basal as well as cAMP- and calcium-induced transcription. Activation of CREB occurs by phosphorylation on serine-119 stimulating its transactivating potency. However, the regulation of CREB-DNA binding by posttranslational modification is not established. In this study, using binding and functional assays, the interaction of CREB with pancreatic islet cell-specific enhancer elements of the rat somatostatin (SMS-UE), glucagon (Glu-G3) and insulin I genes (Ins-E1) was investigated, which share a functional regulatory sequence, PISCES, with islet-specific activity. CREB bound to the SMS-UE. Bacterially expressed recombinant CREB bound equally well to the SMS-UE and to the somatostatin CRE. However, cellular CRE-binding proteins with CREB-like immunoreactivity recognized the SMS-UE markedly less well than the somatostatin CRE suggesting the existence of a posttranslational modification of CREB that alters its binding specificity.

Glucagon gene G3 enhancer: evidence that activity depends on combination of an islet-specific factor and a winged helix protein.

The peptide hormone glucagon is expressed in A cells of the pancreatic islets due to an interaction between multiple regulatory elements within the 5'-flanking region of its gene directing glucagon gene transcription. An A-cell-specific enhancer-like element in the rat glucagon gene, G3, contains two domains, both of which are necessary for G3 activity. Domain A of the G3 element comprises a sequence motif, PISCES, that is also found in control elements of the rat insulin I and somatostatin genes exhibiting cell-specific transcriptional activities distinct from G3. In this study, the nuclear proteins binding to domain B of G3 were characterized. In electrophoretic mobility shift assays using nuclear extracts from a glucagon-producing islet cell line, it was observed that the binding specificity of G3-domain-B-binding proteins is related to that of winged helix proteins supporting the hypothesis that the proteins binding to domain B of G3 may belong to the winged helix protein family of transcription factors. The overexpression of a dominant-negative winged helix protein mutant (derived from HNF-3) virtually abolished the transcriptional activity of G3 in a glucagon-expressing islet cell line. These results suggest that the unique A-cell-specific basal transcriptional activity of the glucagon G3 element depends on a combination of at least two proteins, the islet specific PISCES-binding protein and a more widely expressed winged helix protein.

Female genital schistosomiasis of the lower genital tract: prevalence and disease-associated morbidity in northern Tanzania.

Female genital schistosomiasis (FGS) is a neglected disease manifestation of schistosomiasis. A cross-sectional study was carried out to assess in a schistosomiasis-endemic area the proportion of women affected by FGS of the lower reproductive tract and to compare the frequency of symptoms and signs possibly associated with FGS between women with proven FGS (n=134), endemic referents (n=225, women living in an endemic site), and referents (n=75, women living in a nonendemic site). Urinary schistosomiasis was diagnosed in 36% (239/657) and FGS in 37% (134/359) of the women. Cervical lesions occurred in 75% of the FGS cases, in 48% of endemic referents, and in 36% of nonendemic referents. The high prevalence of FGS in all age groups and the high levels of pathologic cervical alterations such as swollen and disrupted epithelium support the hypothesis that FGS might be a risk factor for the transmission of human immunodeficiency virus.

Distinct properties of the cAMP-responsive element of the rat insulin I gene.

The cAMP response element (CRE)-binding transcription factor CREB can mediate induction of gene transcription in response to calcium as well as to cAMP. Since the rat insulin I gene 5'-flanking region contains a CRE with an octamer-like motif (TGACGTCC), CREB binding and cAMP/calcium responsiveness of the insulin CRE were investigated. In an electrophoretic mobility shift assay and in Southwestern blot experiments, bacterially expressed recombinant CREB bound to the insulin CRE as it did to the rat glucagon and rat somatostatin gene CREs. However, in nuclear extracts of the pancreatic islet cell line HIT, protein complexes binding to the insulin CRE did not contain proteins with CREB-like immunoreactivity, although these bound to the glucagon and somatostatin CREs. When reporter fusion genes were transfected into HIT cells, the isolated insulin CRE increased basal activity and mediated transcriptional activation by cAMP. However, cAMP stimulation of transcription through the insulin CRE was weak when compared with the response through the glucagon and somatostatin CREs. Furthermore, the insulin CRE did not confer responsiveness to membrane depolarization and calcium influx, in contrast to the glucagon and somatostatin CREs. These results demonstrate that the functional properties of the rat insulin I gene CRE are different from those of the rat glucagon and somatostatin CREs which may be explained by a distinct pattern of nuclear protein binding and suggest the existence of post-translational mechanisms that decrease the binding of cellular CREB to the insulin CRE.