The evidence that the DNC (SLC25A19) is not the mitochondrial deoxyribonucleotide carrier

We review the evidence that the function of the SLC25A19 gene product, previously identified as the mitochondrial deoxyribonucleotide carrier (DNC), is actually the transport of thiamine pyrophosphate. This evidence comes from enzyme kinetics, homologous yeast protein alignments, gene knockout studies, and clinical samples from Amish Microcephaly patients. This diverse body of evidence consistently points to the conclusion that SLC25A19 is not the true mitochondrial DNC gene. The identification of the correct mitochondrial DNC is important for research on the genetic diseases of mitochondrial DNA maintenance and the toxicity experienced by many HIV patients undergoing antiretroviral therapy involving nucleoside analogs.     

Duck Productivity in Restored Species-Rich Native and Species-Poor Non-Native Plantings

Conservation efforts to increase duck production have led the United States Fish and Wildlife Service to restore grasslands with multi-species (3-5) mixtures of introduced cool season vegetation often termed dense nesting cover (DNC). The effectiveness of DNC to increase duck production has been variable, and maintenance of the cover type is expensive. In an effort to decrease the financial and ecological costs (increased carbon emissions from plowing and reseeding) of maintaining DNC and provide a long-term, resilient cover that will support a diversity of grassland fauna, restoration of multi-species (16-32) plantings of native plants has been explored. We investigated the vegetation characteristics, nesting density and nest survival between the 2 aforementioned cover types in the Prairie Pothole Region of North Dakota, USA from 2010–2011 to see if restored-native plantings provide similar benefits to nesting hens as DNC. We searched 14 fields (7 DNC, 271 ha; and 7 restored native, 230 ha) locating 3384 nests (1215 in restored-native vegetation and 2169 in DNC) in 2010-2011. Nest survival was similar between cover types in 2010, while DNC had greater survival than native plantings in 2011. Densities of nests adjusted for detection probability were not different between cover types in either year. We found no structural difference in vegetation between cover types in 2010; however, a difference was detected during the late sampling period in 2011 with DNC having deeper litter and taller vegetation. Our results indicate restored-native plantings are able to support similar nesting density as DNC; however, nest survival is more stable between years in DNC. It appears the annual variation in security between cover types goes undetected by hens as hens selected cover types at similar levels both years.     

A dielectric-sensitive fluorescent DNA probe for monitoring polarities on the interior of a DNA-binding protein

We report a novel fluorescent nucleobase, (DNC)U, for the mapping of the local dielectric properties around DNA. Fluorescence spectra of the nucleoside d(DNC)U showed a significant shift depending on the solvent polarity. This property was used to determine the dielectric constants of the interior of DNA-binding proteins, such as DNA polymerases. We were able to demonstrate that the DNA-binding region of the Klenow fragment is a lower polarity site, as shown by the blue shift of the fluorescence peak originating from (DNC)U.     

Morphometric analysis of AgNORs in nonkeratinizing carcinoma and adjacent normal epithelia in the nasopharynx

OBJECTIVE: To evaluate the proliferative activity of different types of nonkeratinizing carcinoma and adjacent normal epithelia in the nasopharynx by the quantitative assessment of argyrophilic nucleolar organizer region (AgNOR) proteins. STUDY DESIGN: Silver staining of nucleolar organizer regions (NORs) was applied to 70 paraffin sections of nonkeratinizing carcinoma in nasopharyngeal biopsies. Fifty-four of the 70 cases had differentiated nonkeratinizing carcinoma (DNC), and the remaining 16 had undifferentiated carcinoma (UC). Nineteen of these 70 samples proved to contain, besides carcinoma, normal epithelia (NE), which was used as a control. The epithelial cells and cancer cells were analyzed for their AgNOR features by image cytometric analysis. RESULTS: As compared with normal epithelia, significant differences were found in mean nuclear area, AgNOR count, mean AgNOR area, AgNOR area ratio and AgNOR area/count ratio between NE and DNC (P < .05) and in mean nuclear area, mean AgNOR area and AgNOR area/count ratio between NE and UC (P < .001). Further, the differences in mean nuclear area, mean AgNOR area and AgNOR area/count ratio were statistically significant between DNC and UC. CONCLUSION: The evaluation of AgNORs is a useful histologic assessment of rapidity of cell proliferation in malignant and benign lesions and demonstrated that UC had more rapidly proliferative activity than DNC in this study.     

New techniques in female pudendal somatosensory evoked potential testing

OBJECTIVE: The primary nerves innervating the female genitalia are the dorsal nerve of the clitoris (DNC) and the perineal nerve, which innervate the clitoris and the external genitalia/distal vagina, respectively. We describe two novel electrodiagnostic techniques for evaluating the integrity of these female genital somatosensory pathways. SUBJECTS AND METHODS: Seventy-seven healthy women (mean age 29.3 years) were enrolled for this study. We performed DNC somatosensory evoked potentials (SEPs), stimulating through self-adhesive disk electrodes on either side of the clitoris. Perineal nerve SEPs were evoked through a vaginal probe. Cortical responses were measured through cup electrodes affixed to the scalp at Cpz and Fpz. Stimulus parameters were duration 0.1 ms, frequency 4.1 Hz, filters 5-5,000 Hz, at three times sensory threshold. RESULTS: DNC and perineal nerve SEPs from both the right and left sides were reproducible and easily discerned. The mean P1 latencies were: right DNC 39.4 ms (SD 2.8 ms), left DNC 39.3 ms (SD 3.3 ms), right perineal nerve 37.8 ms (SD 3.4 ms), left perineal nerve 37.6 ms (SD 3.1 ms). We recorded SEP responses from 90 to 92% of subjects for the DNC, and 69% for the perineal nerve. CONCLUSIONS: We are able to evoke somatosensory potentials from the four primary somatic nerves that mediate female genital cutaneous sensation. In healthy subjects, the DNC responses are robust and maintain laterality. The perineal nerve responses are less consistently obtained, but when recorded, are easily discerned. These preliminary data provide a foundation from which to study female genital innervation, particularly as it applies to sexual function.     

Waterfowl Use of Dense Nesting Cover in the Canadian Parklands

ABSTRACT Dense nesting cover (DNC) has been a conspicuous component of habitat management for upland-nesting ducks for >30 years, but its benefits for nesting ducks have been contentious. During 1994–1999 we monitored 3,058 dabbling duck (Anas spp.) nests in 84 DNC fields located throughout the Canadian Parklands to examine sources of among-field variation in nest density and nesting success. Nest density averaged 1.51 (SE=0.15) nests/ha and overall nesting success was 20.4%, but there was pronounced annual variation in both estimates. Nesting success increased with increasing field size (range = 6–111 ha), but nest density remained constant. Nest density increased with percent wetland habitat within DNC fields and declined with percent perennial cover in the surrounding 2.4 × 2.4-km landscape, but these variables were not important for predicting nesting success. Nest abundance and nesting success roughly doubled in fields seeded with alfalfa (Medicago sativa) or sweet clovers (Melilotus spp.), but there was no benefit from using native as opposed to tame grasses. We recommend that waterfowl managers in the Canadian Parklands establish DNC with alfalfa in large fields in landscapes with abundant wetlands but minimal competing cover.     

Stable expression of a recombinant sodium-dependent, pyrimidine-selective nucleoside transporter (CNT1) in a transport-deficient mouse leukemia cell line.

Previous studies of nucleoside transport in mammalian cells have identified two types of activities: the equilibrative nucleoside transporters and concentrative, Na+-nucleoside cotransporters. Characterization of the concentrative nucleoside transporters has been hampered by the presence in most cells and tissues of multiple transporters with overlapping permeant specificities. With the recent cloning of cDNAs encoding rat and human members of the concentrative nucleoside transporter (CNT) family, it is now possible to study the concentrative transporters in isolation by use of functional expression systems. We report here the isolation of a nucleoside transport-deficient subline of L1210 mouse leukemia (L1210/DNC3) that is a suitable recipient for stable expression of cloned nucleoside transporter cDNAs. We have used L1210/DNC3 as the recipient in gene transfer studies to develop a stable cell line (L1210/DU5) that produces the recombinant concentrative nucleoside transporter with selectivity for pyrimidine nucleosides (CNT1) that was initially identified in rat intestine (Q.Q. Huang, S.Y. Yao, M.W. Ritzel, A.R.P. Paterson, C.E. Cass, and J.D. Young. 1994. J. Biol. Chem. 269: 17,757-17,760). L1210/DU5 was used to examine the permeant selectivity of recombinant rat CNT1 by comparing a series of nucleoside analogs with respect to (i) inhibition of inward fluxes of [3H]thymidine, (ii) initial rates of transport of 3H-analog, and (iii) cytotoxicity to L1210/DU5 versus the parental transport-deficient cell line. By all three criteria, recombinant CNT1 transported 5-fluoro-2'-deoxyuridine and 5-fluorouridine well and cytosine arabinoside poorly. Although some purine nucleosides (2'-deoxyadenosinedeoxyadeno-2'-deoxyadenosine, 7-deazaadenosine) were potent inhibitors of CNT1, they were poor permeants when uptake was measured directly by analysis of isotopic fluxes or indirectly by comparison of cytotoxicity ratios. We conclude that comparison of analog cytotoxicity to L1210/DU5 versus L1210/DNC3 is a reliable indirect predictor of transportability, suggesting that cytotoxicity assays with a panel of such cell lines, each with a different recombinant nucleoside transporter, would be a valuable tool in the development of antiviral and antitumor nucleoside analogs.     

Localization of ribosomal cistrons at the ultrastructural level by in situ hybridization technique

In situ hybridization with 3H 28 S ribosomal RNA at the ultrastructural level shows labelling exclusively over the nucleolus and specifically over the dense nucleolar component (DNC). These findings clearly reveal that the ribosomal cistrons are located in the DNC of the nucleolus.     

New techniques in female pudendal somatosensory evoked potential testing

Objective—The primary nerves innervating the female genitalia are the dorsal nerve of the clitoris (DNC) and the perineal nerve, which innervate the clitoris and the external genitalia/distal vagina, respectively. We describe two novel electrodiagnostic techniques for evaluating the integrity of these female genital somatosensory pathways.

Subjects and methods—Seventy-seven healthy women (mean age 29.3 years) were enrolled for this study. We performed DNC somatosensory evoked potentials (SEPs), stimulating through self-adhesive disk electrodes on either side of the clitoris. Perineal nerve SEPs were evoked through a vaginal probe. Cortical responses were measured through cup electrodes affixed to the scalp at Cpz and Fpz. Stimulus parameters were duration 0.1?ms, frequency 4.1?Hz, filters 5–5,000?Hz, at three times sensory threshold.

Results—DNC and perineal nerve SEPs from both the right and left sides were reproducible and easily discerned. The mean P1 latencies were: right DNC 39.4?ms (SD 2.8?ms), left DNC 39.3?ms (SD 3.3?ms), right perineal nerve 37.8?ms (SD 3.4?ms), left perineal nerve 37.6?ms (SD 3.1?ms). We recorded SEP responses from 90 to 92% of subjects for the DNC, and 69% for the perineal nerve.

Conclusions—We are able to evoke somatosensory potentials from the four primary somatic nerves that mediate female genital cutaneous sensation. In healthy subjects, the DNC responses are robust and maintain laterality. The perineal nerve responses are less consistently obtained, but when recorded, are easily discerned. These preliminary data provide a foundation from which to study female genital innervation, particularly as it applies to sexual function.     

Equilibrium water contents of cellulose films determined via solvent exchange and quartz crystal microbalance with dissipation monitoring

Model cellulose surfaces have attracted increasing attention for studying interactions with cell wall matrix polymers and as substrates for enzymatic degradation studies. Quartz crystal microbalance with dissipation monitoring (QCM-D) solvent exchange studies showed that the water content of regenerated cellulose (RC) films was proportional to the film thickness (d) and was consistent with about five water molecules per anhydroglucose unit. Sulfated nanocrystalline cellulose (SNC) and desulfated nanocrystalline cellulose (DNC) films had comparable water contents and contained about five times more water than RC films. A cellulase mixture served as a probe for studies of substrate accessibility and degradation. Cellulase adsorption onto RC films was independent of d, whereas degradation times increased with d. However, adsorption onto SNC and DNC films increased with d, whereas cellulase degradation times for DNC films were independent of studied d. Enhanced access to guest molecules for SNC and DNC films revealed they are more porous than RC films.     

THE TOXICITY OF SOME INSECTICIDES TO FIFTH-INSTAR NYMPHS OF THE DESERT LOCUST

The toxicities of the insecticides γ-BHC, diazinon, dieldrin, DNC, heptachlor and parathion to fifth-instar hoppers of the desert locust were determined. Parathion and dieldrin were the most toxic, followed in descending order by heptachlor, γ-BHC, diazinon and DNC. The resistance of the hoppers to γ-BHC and DNC changed appreciably with age within the instar.
The effects of some factors which might influence the resistance of the insects were examined and are discussed.     

Curcumin confers protection to irradiated THP-1 cells while its nanoformulation sensitizes these cells via apoptosis induction

Protection against ionizing radiation (IR) and sensitization of cancer cells to IR are apparently contrasting phenomena. However, curcumin takes on these contrasting roles leading to either protection or enhanced apoptosis in different irradiated cells. Here we studied whether pretreatment with free curcumin or a novel dendrosomal nanoformulation of curcumin (DNC) could exert protective/sensitizing effects on irradiated THP-1 leukemia cells. We employed assays including MTT viability, clonogenic survival, DNA fragmentation, PI/Annexin V flow cytometry, antioxidant system (ROS, TBARS for lipid peroxidation, 8-OHdG and γH2AX for DNA damage, glutathione, CAT and GPx activity, enzymes gene expression), ELISA (NF-κB and Nrf2 binding, TNF-α release), caspase assay, siRNA silencing of caspase-3, and western blotting to illustrate the observed protective role of curcumin in comparison with the opposite sensitizing role of its nanoformulation at a similar 10 μM concentration. The in vivo relevance of this concentration was determined via intraperitoneal administration in mice. Curcumin significantly enhanced the antioxidant defense, while DNC induced apoptosis and reduced viability as well as survival of irradiated THP-1 cells. Nrf2 binding showed an early rise and fall in DNC-treated cells, despite a gradual increase in curcumin-treated cells. We also demonstrated that DNC induced apoptosis in THP-1 cells via caspase-3 activation; whereas in combination with radiation, DNC alternatively employed a caspase-independent apoptosis pathway involving cytochrome c release from mitochondria.     

Three descending interneurons reporting deviation from course in the locust

The DNI, DNM and DNC descending interneurons all have very similar properties and are each at the convergence of visual, ocellar, wind-hair and other mechanoreceptor inputs. The 3 neurons respond almost exclusively to movement of the animal in space about its three axes of rotation. All are spatially and directionally selective. Movements in the preferred sense produce increasingly strong responses with amplitude and absolute position, while movements in the antipreferred sense usually elicit no response at all. Movements in the preferred sense, but towards, rather than away from, the normal flying position start to produce responses only as the animal approaches the normal flight position. The neurons function as feature detectors, responding only to specific sorts of deviation from course. DNI, DNM and DNC differ from one another principally in their directionality. DNI responds optimally to a diving banked turn to the ipsilateral side, DNM to downwards pitch, and the DNC to a diving banked turn to the contralateral side. The DN neurons contribute to the production of steering manoeuvres. They appear to be representatives of a larger class of descending interneurons bringing exteroceptive sensory input to the thoracic locomotory neuropil. The occurrence of this class of units in locusts and other insects is discussed.     

Selective labelling of melittin with a fluorescent dansylcadaverine probe using guinea-pig liver transglutaminase.

Melittin, a C-terminal glutamine peptide, incorporated the fluorescent probe monodansylcadaverine (DNC) when catalysed by guinea-pig liver transglutaminase and Ca2+, as determined by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). A 1:1 adduct DNC-melittin was identified in which a single glutamine residue out of two, i.e. Gln25, acts as acyl donor. Incubation of melittin with transglutaminase in the absence of DNC originated high molecular mass complexes indicative that the peptide lysine residue can act as an acyl acceptor. The DNC-melittin was about 3 times more active in the lysis of red cell membranes than native melittin. Fluorescence study of the labelled melittin in the submicromolar range where it is active on cells showed that while totally exposed to solvent in methanol solution, both Trp and dansyl groups are buried in buffer solution. This strongly suggests that DNC-melittin is self-associated and indeed more active than the native melittin in the same conditions.     

Nanoformulation of curcumin protects HUVEC endothelial cells against ionizing radiation and suppresses their adhesion to monocytes: potential in prevention of radiation-induced atherosclerosis

Objectives

To investigated the potential of a novel dendrosomal nanoformulation of curcumin (DNC) in blocking radiation-induced changes in irradiated human umbilical vein endothelial cells (HUVECs), and their adhesion to human THP-1 monocytoid cells.

Results

Co⁶⁰ gamma rays reduced viability, raised the expression of adhesion molecules, ICAM-1, VCAM-1 and E-selectin (mRNA and protein), augmented the adhesion of THP-1 cells to HUVECs, activated NF-κB binding, increased the release of pro-inflammatory cytokines (IL-6, IL-8 and MCP-1) and induced oxidative damage (reduced glutathione declined, while 8-OHdG and TBARS increased). 5 µM DNC significantly inhibited these radiation-induced changes, activated the Nrf-2 pathway, and effectively suppressed THP-1 adhesion to HUVECs, implicating p38 MAPK signaling.

Conclusion

DNC treatment is a potential preventive method against inflammation and vascular damage from ionizing radiation.

     

A caspase-8-independent component in TRAIL/Apo-2L-induced cell death in human rhabdomyosarcoma cells

Tumor necrosis factor related apoptosis inducing ligand (TRAIL) belongs to the Tumor necrosis factor (TNF) family of death-inducing ligands, and signaling downstream of TRAIL ligation to its receptor(s) remains to be fully elucidated. Components of the death-inducing signaling complex (DISC) and TRAIL signaling downstream of receptor activation were examined in TRAIL - sensitive and -resistant models of human rhabdomyosarcoma (RMS). TRAIL ligation induced DISC formation in TRAIL-sensitive (RD, Rh18, Rh30) and TRAIL-resistant RMS (Rh28, Rh36, Rh41), with recruitment of FADD and procaspase-8. In RD cells, overexpression of dominant-negative FADD (DNFADD) completely abolished TRAIL-induced cell death in contrast to dominant-negative caspase- 8 (DNC8), which only partially inhibited TRAIL-induced apoptosis, growth inhibition, or loss in clonogenic survival. DNC8 did not inhibit the cleavage of Bid or the activation of Bax. Overexpression of Bcl-2 or Bcl-xL inhibited TRAIL-induced apoptosis, growth inhibition, and loss in clonogenic survival. Bcl-2 and Bcl-xL, but not DNC8, inhibited TRAIL-induced Bax activation. Bcl-xL did not inhibit the early activation of caspase-8 (<4 h) but inhibited cleavage of Bid, suggesting that Bid is cleaved downstream of the mitochondria, independent of caspase-8. Exogenous addition of sphingosine also induced activation of Bax via a caspase-8-and Bid-independent mechanism. Further, inhibition of sphingosine kinase completely protected cells from TRAIL-induced apoptosis. Data demonstrate that in RMS cells, the TRAIL signaling pathway circumvents caspase-8 activation of Bid upstream of the mitochondria and that TRAIL acts at the level of the mitochondria via a mechanism that may involve components of the sphingomyelin cycle.     

Modelling geographic patterns of population density of the white‐tailed deer in central Mexico by implementing ecological niche theory

Conservation and management of species require basic knowledge on their geographic distribution and abundance. Here, we propose a novel approach, based on the theory of the ecological niche, to model the spatial patterns of the white‐tailed deer Odocoileus virginianus population density in two regions of central Mexico (Balsas Basin and Tehuacán‐Cuicatlán Valley). We used an ecological niche model to generate binary geographic distribution maps of the white‐tailed deer in each region based on occurrence data and a set of environmental variables. Then, the centroid of the distributions was calculated in ecological space (niche centroid) and the multidimensional Euclidian ecological distance of each pixel to the niche centroid was estimated. Finally, for each region the distance to the niche centroid (DNC) was regressed against 14 independent occurrence points in each site containing white‐tailed deer density information to determine the function describing the DNC‐density relationship, which was used to generate maps describing the distribution of white‐tailed deer density. Our results indicated an inverse DNC‐density relationship in both regions (Balsas Basin: r² = 0.90 and Tehuacán‐Cuicatlán: r² = 0.76) that was validated via bootstrapping resulting in a predicting capacity of near 62% for Balsas Basin and 65% for Tehuacán‐Cuicatlán Valley. Our results suggest that the distance to the niche centroid method is a robust, science‐based correlative approach that resulted useful to predict the population density of the white‐tailed deer in a spatially explicit fashion. The proposed approach is suitable for predicting the distribution of density for white‐tailed deer for which occurrence data with accompanying density information exists, but relative abundance can also be estimated when no abundance data are available.