Use of cervicovaginal fluid for the identification of biomarkers for pathologies of the female genital tract

Cervicovaginal fluid has an important function in the homeostasis and immunity of the lower female genital tract. Analysis of the cervicovaginal fluid proteome may therefore yield important information about the pathogenesis of numerous gynecological pathologies. Additionally, cervicovaginal fluid has great potential as a source of biomarkers for these conditions.     

Angiotensin II and angiotensin-(1-7) decrease sFlt1 release in normal but not preeclamptic chorionic villi: an <Emphasis Type="Italic">in vitro</Emphasis> study

Background  

During preeclampsia, placental angiogenesis is impaired. Factors released from the placenta including vascular endothelial growth factor (VEGF), placental growth factor (PLGF), soluble VEGF receptor 1 (sFlt1), and soluble endoglin (sEng) are regulatory molecules of placental development and function. While the renin angiotensin system has been shown to regulate angiogenic factors in other research fields, these mechanisms have not been extensively studied during pregnancy.     

Investigation of the effect of Interleukin-1 receptor antagonist (IL-1ra) on markers of inflammation in non-ST elevation acute coronary syndromes (The MRC-ILA-HEART Study)

Background

Point of care testing (PoCT) may be a useful adjunct in the management of chronic conditions in general practice (GP). The provision of pathology test results at the time of the consultation could lead to enhanced clinical management, better health outcomes, greater convenience and satisfaction for patients and general practitioners (GPs), and savings in costs and time. It could also result in inappropriate testing, increased consultations and poor health outcomes resulting from inaccurate results. Currently there are very few randomised controlled trials (RCTs) in GP that have investigated these aspects of PoCT.

Design/Methods

The Point of Care Testing in General Practice Trial (PoCT Trial) was an Australian Government funded multi-centre, cluster randomised controlled trial to determine the safety, clinical effectiveness, cost effectiveness and satisfaction of PoCT in a GP setting. The PoCT Trial covered an 18 month period with the intervention consisting of the use of PoCT for seven tests used in the management of patients with diabetes, hyperlipidaemia and patients on anticoagulant therapy. The primary outcome measure was the proportion of patients within target range, a measure of therapeutic control. In addition, the PoCT Trial investigated the safety of PoCT, impact of PoCT on patient compliance to medication, stakeholder satisfaction, cost effectiveness of PoCT versus laboratory testing, and influence of geographic location.

Discussion

The paper provides an overview of the Trial Design, the rationale for the research methodology chosen and how the Trial was implemented in a GP environment. The evaluation protocol and data collection processes took into account the large number of patients, the broad range of practice types distributed over a large geographic area, and the inclusion of pathology test results from multiple pathology laboratories. The evaluation protocol developed reflects the complexity of the Trial setting, the Trial Design and the approach taken within the funding provided. The PoCT Trial is regarded as a pragmatic RCT, evaluating the effectiveness of implementing PoCT in GP and every effort was made to ensure that, in these circumstances, internal and external validity was maintained.

Trial Registration

12612605000272695     

An improved formulation of the zirconyl hematoxylin stain for acidic mucins

Zirconyl hematoxylin stains acidic mucins darkly and specifically using a solution of 100 mg hematoxylin, 5 ml ethanol, 5 ml 0.5% sodium iodate, 400 mg zirconyl chloride octahydrate, and 30 ml 25% aqueous glycerol. The stain is especially advantageous for studying goblet cells and Paget cells.     

Selective, high-resolution fluorescence imaging of mitochondrial Ca2+ concentration.

We have developed a digital image processing technique based on highpass filtering of microfluorimetric images for selective transmission of fine image details corresponding to mitochondria. This technique enabled the detection of the mitochondrial calcium signals with high selectivity, simultaneously with the cytosolic calcium signal. The validity of this technique was supported in primary cultures of rat brain capillary endothelial cells loaded with X-rhod-1 by the results that (i) inhibition of the mitochondrial Ca2+ uptake by discharging the mitochondrial membrane potential selectively abolished the transient of the highpass filtered signal evoked by ATP, and (ii) CGP-37157, a selective blocker of the mitochondrial Na+/Ca2+ exchanger, increased the peak amplitude of highpass filtered (mitochondrial) Ca2+ transients and caused a sustained plateau. The highpass filtering technique enabled the analysis of the mitochondrial Ca2+ transients in high temporal resolution. We found a uniform and monophasic rise of [Ca2+] in the mitochondrial population of the cell, following the cytosolic [Ca2+] with a delay at onset and peak. The introduced highpass filtering technique is a powerful tool in the high spatial and temporal resolution analysis of mitochondrial calcium transients, and it could be especially important in specimens where genetically targeted probes fail.     

NaCl-preferring NZB/B1NJ mice and NaCl-avoiding CBA/J mice have similar amiloride inhibition of chorda tympani responses to NaCl

Integrated chorda tympani nerve responses to NaCl were studied in two mouse strains, an NaCl-preferring NZB/B1NJ and an NaCl-avoiding CBA/J. The NaCl responses of both strains had similar magnitude and were suppressed by amiloride to a similar extent. This suggests that peripheral gustatory responsiveness to NaCl is not the only mechanism underlying mouse strain variation in NaCl acceptance.      

Organization of fusicoccin receptor in higher plant plasma membrane: relationship between affinity and molecular mass

Higher plant plasma membranes carry receptors of different affinity for the phytotoxin fusicoccin. Reception of fusicoccin involves proteins belonging to the highly conserved 14-3-3 family, but the complete structure of the fusicoccin receptor (FCR) is unknown. Using radiation inactivation analysis, we estimated the molecular masses of low-affinity and high-affinity FCR at 63 +/- 7 and 130 +/- 15 kD, respectively. The dose dependences of receptor inactivation indicate that microsomal specimens contain "silent" FCRs of 420 +/- 90 kD in amounts commensurate with that of the active FCRs. Both low- and high-affinity FCRs are inactivated by hydrolytic enzymes from the outer surface of the plasma membrane, and impairment of protoplast integrity causes an irreversible transition of the low-affinity binding site into the high-affinity one. A scheme is proposed for the organization of different types of FCR in the plasma membrane, implying that the membrane affinity for fusicoccin reflects the interaction between proteins in the FCR complex.     

INDUCTION OF ERYTHROID MATURATION BY DIMETHYL SULFOXIDE IN FRIEND LEUKEMIC CELLS

Enhancement of the erythroid maturation in Friend virus-induced leukemic cells has been examined in vitro by the treatment with dimethyl sulfoxide (DMSO). Although the cell growth was inhibited in the medium containing 2% DMSO, many cells remained viable for a week. By the 3rd day of the culture, the cells treated with DMSO became more strongly agglutinated by phytohemagglutinin than the cells incubated without DMSO. Mouse erythrocyte membrane-specific antigens were also detectable at the 4th day. At the 8th day of the culture hemoglobin synthesis was apparently demonstrated in the cells treated with DMSO, which could not be seen in the untreated cells. Maturation or differentiation along the erythroid pathway in Friend leukemic cells by DMSO is discussed on these markers.     

Molecular genetics of <Emphasis Type="Italic">Cicindela (Cylindera) terricola</Emphasis> and elevation of <Emphasis Type="Italic">C. lunalonga</Emphasis> to species level,with comments on its conservation status

Cicindela (Cylindera) terricola Say is one of the most widespread and variable species of Nearctic Cicindelidae with six recognized subspecies. Cicindela t. lunalonga Schaupp (1884) is known from few museum specimens collected prior to 1979. The goal of this study was to resolve the uncertain taxonomic status of C. t. lunalonga using mitochondrial DNA analysis of cytochrome b and cytochrome oxidase subunit I and determine its conservation status. In phylogenetic reconstruction using distance and parsimony methods, all members of the terrricola group were recovered as monophyletic and embedded within outgroup species of the subgenus Cylindera, while C. lunalonga was recovered as sister to all other members of the C. terricola clade. Cicindela lunalonga exhibited an exceptionally high (mean of 6.36%) pairwise sequence divergence for both genes against all C. terricola surveyed. For the cytochrome oxidase subunit I alone the pairwise divergence was 3.9–4.8% (4.58% avg.). The lowest divergences were between C. lunalonga and C. terricola subspecies of the American southwest (C. t. cinctipennis and C. t. kaibabensis), rather than with the closest geographic neighbors (C. t. imperfecta). We conclude that based on strict monophyly and pairwise sequence divergence, C. lunalonga is a distinct species. Our study of museum specimens and extensive field surveys suggest this species has been extirpated from all sites in the San Joaquin Valley and perhaps all but one of the historic sites throughout its range. Thus, it should be considered for Federal listing as an endangered species.     

Population genetics of three at-risk tiger beetles Habroscelimorpha dorsalis dorsalis,H. d. media,and Ellipsoptera puritana

Many tiger beetles (Family Cicindelidae) are critically imperiled due to their dependence on small patches of suitable habitat that are frequently threatened by natural and anthropogenic disturbances. In the eastern United States, conservation of three tiger beetles - Habroscelimorpha dorsalis dorsalis, H. dorsalis media, and Ellipsoptera puritana - has been inhibited by the absence of population genetic information that is needed for effective recovery planning and potential reintroductions. Using microsatellite panels, we performed population genetic analyses and compared patterns in diversity and differentiation within and between taxa. Nearly all collections of the three taxa had less observed heterozygosity than expected under Hardy-Weinberg Equilibrium, and there was a strong latitudinal gradient in genetic diversity in H. d. dorsalis distributed along the eastern and western shores of the Chesapeake Bay. We also found clear spatial patterns of genetic differentiation which reflected strong isolation-by-distance within all three taxa and between collections of H. d. dorsalis and H. d. media. However, there was evidence of admixture in current (mouth of the Chesapeake Bay) and former (coastal New Jersey) contact zones of H. d. dorsalis and H. d. media. Taken together, our study suggests that relatively few adult tiger beetles may maintain many populations, and that gene flow among nearby habitat patches is common in all three taxa – a characteristic that may help tiger beetles persist in dynamic coastal environments. Results of our analyses can be used to support conservation and management by identifying the spatial scale of metapopulation connectivity and locating populations at the greatest risk of extirpation.