High level of divergence of male-reproductive-tract proteins, between Drosophila melanogaster and its sibling species, D. simulans

We compared male-reproductive-tract polypeptides of Drosophila melanogaster and D. simulans by using two-dimensional gel electrophoresis. Approximately 64% of male-reproductive-tract polypeptides were identical between two randomly chosen isofemale lines from these two species, compared with 83% identity for third-instar imaginal wing-disc polypeptides. Qualitatively similar differences were found between reproductive tracts and imaginal discs when D. sechellia was compared with D. melanogaster and with D. simulans. When genic polymorphism was taken into account, approximately 10% of male- reproductive-tract polypeptides were apparently fixed for different alleles between D. melanogaster and D. simulans; this proportion is the same as that found for soluble enzymes by one-dimensional gel electrophoresis. Strikingly, approximately 20% of male-reproductive- tract polypeptides of either D. melanogaster or D. simulans had no detectable homologue in the other species. We propose that proteins of the Drosophila male reproductive tract may have diverged more extensively between species than have other types of proteins and that much of this divergence may involve large changes in levels of polypeptide expression.      

Microelectrode studies of the active Na transport pathway of frog skin

When the outer surface of short-circuited frog skin was penetrated with microelectrodes, stable negative potentials that averaged near -100 mV were recorded consistently, confirming the results of Nagel (W. Nagel. 1975. Abstracts of the 5th International Biophysics Congress, Copenhagen. P-147.). The appearance of these stable potentials, V(O), concurrent with the observations that (a) a high resistance outer barrier R(O) accounting for approximately 75 percent or more of the transcellular resistance of control skins had been penetrated and that (b) 10(-5) M amiloride and reduced [Na] outside caused the values of V(O) to increase towards means value near -130 mV while the values of percent R(O) increased to more than 90 percent. It was of relationships were the same as the values of E(1) observed in studies of the current-voltage relationships were the same as the values of E’(1) defined as the values of voltage at the inner barrier when the V(O) of the outer barrier was reduced to zero by voltage clamping of the skins. Accordingly, these data are interpreted to mean that the values of E(1), approximately 130 mV, represent the E(Na) of the sodium pump at the inner barrier. 2,4-DNP was observed to decrease the values of transepithelial voltage less than E(1) the V(O) was negative. These data can be interpreted with a simple electrical equivalent circuit of the active sodium transport pathway of the frog skin that includes the idea that the outer membrane behaves as an electrical rectifier for ion transport.     

The biogeographic history of Phoenicurus redstarts reveals an allopatric mode of speciation and an out-of-Himalayas colonization pattern

Montane areas host high levels of diversity and endemism, and these features are tied to habitat stratification along an elevational gradient. As such, montane areas are often thought of as model systems in which sympatric speciation can occur. To test this idea, we selected Phoenicurus redstarts, an avian genus with an extensive distribution across Eurasia, as well as Northwest Africa; nine of the 14 species in the genus have distributions which include the Himalayas. We used sequences of the mtDNA ND2 and cytochrome-b genes and intron 9 of the Z chromosome specific ACO1 gene to reconstruct a phylogeny of the genus. The resulting trees were used to reconstruct a biogeographic history of Phoenicurus, and to date diversification events. We also analysed the relationship between node age and sympatry to determine the geographic mode of speciation in the genus. Our data suggest a very late Miocene, Himalayan origin for Phoenicurus. Diversification and colonization of other parts of Eurasia, as well as Northwest Africa, continued through the Pleistocene, with a rapid pulse of speciation in the late Pliocene. Allopatric speciation was the dominant mode of speciation in Phoenicurus, despite extensive distributional overlaps in the Himalayas where ecological conditions are amenable to speciation in sympatry. Our results, along with several other studies, suggest an emerging pattern where the Himalayas served as a source area for montane specialist avian lineages that subsequently colonized other Palaearctic regions.     

Rapid fluorescent visualization of multiple NAD(P)H oxidoreductases in homogenate,permeabilized cells,and tissue slices

Intracellular NAD(P)H oxidoreductases are a class of diverse enzymes that are the key players in a number of vital processes. The method we present and validate here is based on the ability of many NAD(P)H oxidoreductases to reduce the superoxide probe lucigenin, which is structurally similar to flavins, to its highly fluorescent water-insoluble derivative dimethylbiacridene. Two modifications of the method are proposed: (i) an express method for tissue homogenate and permeabilized cells in suspensions and (ii) a standard procedure for cells in culture and acute thin tissue slices. The method allows one to assess, visualize, and localize, using fluorescent markers of cellular compartments, multiple NADH and NADPH oxidoreductase activities. The application of selective inhibitors (e.g., VAS2870, a NOX2 inhibitor; plumbagin, a NOX4 inhibitor) allows one to distinguish and compare specific NAD(P)H oxidoreductase activities in cells and tissues and to attribute them to known enzymes. The method is simple, rapid, and flexible. It can be easily adapted to a variety of tasks. It will be useful for investigations of the role of various NAD(P)H oxidoreductases in a number of physiological and pathophysiological processes.     

Fluorescence characteristics and photophysical parameters of light-harvesting chlorophyll <Emphasis Type="Italic">a/b</Emphasis> complex aggregates

Fluorescence characteristics and molecular photophysical parameters of light-harvesting chlorophyll a/b complexes isolated from pea were studied in relation to their aggregation state. The aggregate size was varied by changing the Triton X-100 concentration from 0 to 0.23 mM at a chlorophyll concentration of 2.45 μg/ml. Molecular photophysical parameters were determined with laser fluorimetry. Dispersion of large aggregates into smaller ones drastically decreased the intensity of low-temperature (77 K) fluorescence at 700 nm, reduced the singlet-singlet annihilation rate by more than two orders of magnitude, and prolonged the fluorescence lifetime from 0.16 to 3.2 ns.     

Experience on clinical application of positron emission tomography with a new radiopharmaceutical preparation sodium 11C-butyrate in differential diagnosis of pathological volume in brain formation

The study was aimed to investigate the possibilities of the application of a new radiopharmaceutical--"Sodium butyrate, [1-(11)C]" (11C-Butyrate) for differential diagnosis of pathological volume brain formations. Positron emission tomography (PET) ("Ecat Exact 47", Siemens) of the brain was performed in 24 patients (14 male & 10 female, aged 19-56). The scanning was carried out as static scan 4-5 min after intravenous administration of 250-350 Mbq 11C-Butyrate and proceeded within 15-20 min. In addition PET with 18F-FDG was performed in all patients. The data had been verified histologically and morphologically. In 15 out of 24 patients the brain malignant tumor has been discovered, in 4 patients--benign tumors, in 5--postoperative cysts. Results of the studies showed that the ratio tumor/normal tissue in case of 11C-Butyrate and 18F-FDG application proved to be comparable for all studied hystological types of tumors. Malignant tumors in these cases had been visualized as a hot sports-fokci of increased uptake 11C-Butyrate (ratio > 1). The obtained data testify to the possibility of 11C-Butyrate application for differential diagnosis of the pathological volumic brain formation.     

Clinical experience with positron emission tomography with 2-fluorine, 18F-2-deoxy-d-glucose used for the diagnosis of malignant breast neoplasms

The aim of the study was to evaluate sensitivity and specificity of positron emission tomography (PET) and 18F-fluorodesoxyglucose (18F-FDG) in breast cancer diagnosis and to assess tumor dissemination. Sixty two patients were examined: 10 without mammary disease, 10 with fibrous cystic mastopathy, and 42 with breast cancer, which was hystologically verified. PET scanning was recorded in the "Whole body" mode 70-90 min later administration of 370-420 MBq 18F-FDG. It was shown that PET has a high diagnostic accuracy in breast cancer detection. There were no False-positive cases in our investigations. PET scanning in the "Whole body" mode was allowed to assess dissemination of tumor process with high accuracy. Metastatic involvement of local lymph nodes was detected in 82.8% cases and bone metastases--86% in cases.     

Electrically Contacted Bienzyme‐Functionalized Mesoporous Carbon Nanoparticle Electrodes: Applications for the Development of Dual Amperometric Biosensors and Multifuel‐Driven Biofuel Cells

The capping of electron relay units in mesoporous carbon nanoparticles (MPC NPs) by crosslinking of different enzymes on MPC NPs matrices leads to integrated electrically contacted bienzyme electrodes acting as dual biosensors or as functional bienzyme anodes and cathodes for biofuel cells. The capping of ferrocene methanol and methylene blue in MPC NPs by the crosslinking of glucose oxidase (GOx) and horseradish peroxidase (HRP) yields a functional sensing electrode for both glucose and H₂O₂, which also acts as a bienzyme cascaded system for the indirect detection of glucose. A MPC NP matrix, loaded with ferrocene methanol and capped by GOx/lactate oxidase (LOx), is implemented for the oxidation and detection of both glucose and lactate. Similarly, MPC NPs, loaded with 2,2′‐azino‐bis(3‐ethylbenzo­thiazoline‐6‐sulphonic acid), are capped with bilirubin oxidase (BOD) and catalase (Cat), to yield a bienzyme O₂ reduction cathode. A biofuel cell that uses the bienzyme GOx/LOx anode and the BOD/Cat cathode, glucose and/or lactate as fuels, and O₂ and/or H₂O₂ as oxidizers is assembled, revealing a power efficiency of ≈90 μW cm^?2 in the presence of the two fuels. The study demonstrates that multienzyme MPC NP electrodes may improve the performance of biofuel cells by oxidizing mixtures of fuels in biomass.     

Direct intracardiac injection of umbilical cord-derived stromal cells and umbilical cord blood-derived endothelial cells for the treatment of ischemic cardiomyopathy

The development of new therapeutic strategies is necessary to reduce the worldwide social and economic impact of cardiovascular disease, which produces high rates of morbidity and mortality. A therapeutic option that has emerged in the last decade is cell therapy. The aim of this study was to compare the effect of transplanting human umbilical cord-derived stromal cells (UCSCs), human umbilical cord blood-derived endothelial cells (UCBECs) or a combination of these two cell types for the treatment of ischemic cardiomyopathy (IC) in a Wistar rat model. IC was induced by left coronary artery ligation, and baseline echocardiography was performed seven days later. Animals with a left ventricular ejection fraction (LVEF) of ≤40% were selected for the study. On the ninth day after IC was induced, the animals were randomized into the following experimental groups: UCSCs, UCBECs, UCSCs plus UCBECs, or vehicle (control). Thirty days after treatment, an echocardiographic analysis was performed, followed by euthanasia. The animals in all of the cell therapy groups, regardless of the cell type transplanted, had less collagen deposition in their heart tissue and demonstrated a significant improvement in myocardial function after IC. Furthermore, there was a trend of increasing numbers of blood vessels in the infarcted area. The median value of LVEF increased by 7.19% to 11.77%, whereas the control group decreased by 0.24%. These results suggest that UCSCs and UCBECs are promising cells for cellular cardiomyoplasty and can be an effective therapy for improving cardiac function following IC.     

Mitochondrial DNA suggests independent evolutionary history and population decline of the Menzbir’s pipit (Anthus [gustavi] menzbieri)

In this study we use mtDNA ND2 gene (1041 bp) to evaluate the relationship between Menzbir’s (Anthus [gustavi] menzbieri) and Pechora (A. [g.] gustavi) pipits. Menzbir’s pipit is listed in the regional Red Data Book as a distinct, rare species with a small range. We obtained 18 Pechora pipit samples from two localities and 8 Menzbir’s pipit samples from a single locality. Sequences of the two taxa appear reciprocally monophyletic and are separated by 6 substitutions (0.6% divergence). Differences between the taxa explained 62.4% of the variation in our dataset. Differences among individuals within localities explained 34.8%, whereas differences between the two Pechora pipit localities explained only 2.8%. Mismatch distributions suggest that unlike the Pechora pipit localities, which either have experienced recent population growth or sustain a stable population size, the Menzbir’s pipit population may be declining. Our results suggest distinct taxonomic and conservation status for the Menzbir’s pipit.