Decrease in tumor cell contamination and progenitor cell yield in leukapheresis products after consecutive cycles of chemotherapy for breast cancer treatment.
In this retrospective study, we assessed the impact of each of three consecutive cycles of conventional-dose chemotherapy on CD34+ cells, colony-forming units granulocyte-macrophage (CFU-GM), and contaminating breast cancer cells collected in the leukapheresis products of patients with metastatic breast cancer. The patients subsequently underwent high-dose chemotherapy followed by autologous blood progenitor cell transplantation. We analyzed 172 leukapheresis products from 17 patients and have correlated the long-term clinical outcome with tumor cell contamination. The induction chemotherapy regimen consisted of three cycles of cyclophosphamide 750 mg/m2 i.v., epirubicin 100 mg/m2, and 5-fluorouracil (5-FU) 750 mg/m2 i.v., followed by 5 microg/kg body weight of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) daily until leukapheresis was completed. An average of 10 leukapheresis products (three to four collections after each cycle of chemotherapy) were obtained from each patient. Numbers of CD34+ cells, CFU-GM, and mononuclear cells (MNCs) in the leukapheresis products were determined at the time of collection. Aliquots from the same products were frozen and breast cancer cells were detected by immunocytochemistry with a cocktail of anti-cytokeratin antibodies (AE-1, AE-3, CAM 5.2, Keratin 8+18+19) using a standardized immunoalkaline phosphatase method. A minimum of 10(6) cells were examined by light microscopy and by at least two blinded observers. Cells were considered positive when immunostaining was detected in the cytoplasm and on the cell membrane, and cellular morphology was consistent with a malignant phenotype. Of the 172 samples analyzed, 13 of 57 (23%) leukapheresis products collected after cycle I were positive for tumor cells; 3 of 60 (5%) after cycle II; and 4 of 55 (7%) after cycle III. The likelihood of contamination by breast cancer cells after cycle I was significantly higher than after subsequent cycles of chemotherapy (p = 0.0052). Simultaneously, there was a significant decrease in quantity of CD34+ cells and CFU-GM (p < 0.0001 for both comparisons). Our study indicated that leukapheresis products collected after the second or third cycles of induction chemotherapy carry a significantly lower likelihood of tumor cell contamination, albeit the quantity of CD34+ cells or CFU-GM collected was also significantly reduced.


Polarization-insensitive fiber-coupled superconducting-nanowire single photon detector using a high-index dielectric capping layer.
Superconducting-nanowire single photon detectors (SNSPDs) are able to reach near-unity detection efficiency in the infrared spectral range. However, due to the intrinsic asymmetry of nanowires, SNSPDs are usually very sensitive to the polarization of the incident radiation, their responsivity being maximum for light polarized parallel to the nanowire length (transverse-electric (TE) polarization). Here, we report on the reduction of the polarization sensitivity obtained by capping NbN-based SNSPDs with a high-index SiNx dielectric layer, which reduces the permittivity mismatch between the NbN wire and the surrounding area. Experimentally, a polarization sensitivity below 0.1 is obtained both at 1.31 and 1.55 µm, in excellent agreement with simulations.


Sinus floor augmentation at the time of maxillary molar extraction: technique and report of preliminary results.
A technique is described for accomplishing both localized sinus augmentation and guided bone regeneration at the time of maxillary molar extraction. One hundred nine sites were treated in 92 patients. Of these, 102 procedures (94.0%) were successful and 7 (6.0%) were partially successful. Success was defined as the ability to ideally position an implant at least 10 mm in length and 4.8 mm in width without perforating the floor of the sinus or generating an implant fenestration or dehiscence. Partially successful procedures required an additional osteotome sinus lift at the time of implant placement.


Clinical and pathologic features of chronic adriamycin toxicosis in rabbits.
Chronic adriamycin (ADR) intoxication was produced in 50 weanling rabbits by weekly injections of ADR (2.4 mg/kg of body weight) for up to 17 weeks. All ADR-treated rabbits developed prominent alopecia, edema, moderate anemia, and severe emaciation. The frequency and severity of cardiomyopathy were not strongly dose-related, but renal lesions were. The frequency and severity of cardiomyopathic lesions were highest in left ventricular free wall and ventricular septum, intermediate in atria, and lowest in right ventricular free wall. Cardiac lesions increased in frequency and severity basally in left ventricular free wall, ventricular septum, and atria, but not in right ventricular free wall. Nephropathy lesions were selective for inner cortex and constitute a common and important side effect of chronic ADR intoxication in rabbits. Other lesions observed at necropsy were hepatic necrosis, skeletal muscle degeneration, osteodystrophy-associated fractures, peripancreatic fat necrosis, and testicular degeneration and atrophy.


Viral gene expression and provirus load of Orf-A defective FIV in lymphoid tissues and lymphocyte subpopulations of neonatal cats during acute and chronic infections.
Neonatal cats were infected with a wild type (JSY3) or orf-A defective (JSY3DeltaORF-A) feline immunodeficiency virus (FIV) to determine the provirus load and level of viral gene expression at the acute versus chronic stages of infection. FIV DNA in the thymus, lymph node, peripheral blood mononuclear cells (PBMCs) and lymphocyte subpopulations at week 8 post-infection was lower in animals infected with JSY3DeltaORF-A as compared to that of JSY3. At week 16 we observed no significant difference in provirus load between the two groups except for B cells where it was higher in the JSY3 infection. In B cells proviral burden was found to be the same in animals infected with JSY3 for both time points. In the chronic stage, therefore, proviral burden dominates in B cells for JSY3, whereas the level of JSY3DeltaORF-A was lower with comparable values for all lymphocytes at both weeks 8 and 16. Gene expression profiles as measured by real time PCR for gag and rev transcripts revealed decreased levels of JSY3DeltaORF-A mRNAs as compared to that of JSY3. The JSY3 chronic phase infection showed viral gene expression to be higher in B cells relative to CD4+ and CD8+ cells. The presence of viral RNA in CD8 and B cells during the chronic infection implicates active virus replication. Hematological profiles revealed that there was a decline in the number of B cells in JSY3DeltaORF-A-infected cats during the chronic stage of infection while no significant change was observed in animals infected with the wild type virus. Comparative analysis of cell numbers to provirus load and levels of viral transcripts in CD4+ and CD8+, however, did not correlate cell numbers to the levels of viral DNA and gene expression. It remains to be determined whether the relatively high virus burden in B cells as compared to CD4+ and CD8+ cells reflects a role for Orf-A in a shift to B cell virus load during the chronic stage of FIV infection.


Human cases of West Nile Virus infection in north-eastern Italy, 15 June to 15 November 2010.
In 2010, for the third consecutive year, human cases of West Nile virus (WNV) infection, including three confirmed cases of neuroinvasive disease and three confirmed cases of West Nile fever, were identified in north-eastern Italy. While in 2008 and 2009 all human cases of WNV disease were recorded in the south of the Veneto region, cases of WNV disease in 2010 additionally occurred in two relatively small northern areas of Veneto, located outside those with WNV circulation in the previous years. WNV IgG antibody prevalence in blood donors resident in Veneto was estimated as ranging from 3.2 per 1,000 in areas not affected by cases of WNV disease to 33.3 per 1,000 in a highly affected area of the Rovigo province. No further autochthonous human cases of WNV disease were notified in Italy in 2010. The recurrence of human cases of WNV infection for the third consecutive year strongly suggests WNV has become endemic in north-eastern Italy.


Identification of IL-7-producing cells in primary and secondary lymphoid organs using IL-7-GFP knock-in mice.
IL-7 is a cytokine crucial for development and maintenance of lymphocytes and other hematopoietic cells. However, how IL-7-expressing cells are distributed in lymphoid organs is not well known. To address this question, we established and analyzed IL-7-GFP knock-in mice. Thymic epithelial cells (TECs) expressed high GFP levels in the cortex and medulla, as detected with an anti-GFP Ab. Thymic mesenchymal cells also expressed GFP. Flow cytometry analysis suggested that cortical TECs expressed higher GFP levels than did medullary TECs. In bone marrow, immunohistochemistry indicated high levels of GFP in many VCAM-1(+) mesenchymal stromal cells and in some VCAM-1(-) cells. Additionally, half of the VCAM-1(+)CD31(-) stromal cells and some platelet-derived growth factor receptor α(+) stromal cells were GFP(+), as detected by flow cytometry. Moreover, we detected GFP expression in fibroblastic reticular cells in the T cell zone and cortical ridge of lymph nodes. Remarkably, lymphatic endothelial cells (LECs) expressed GFP at high levels within the lymph node medulla, skin epidermis, and intestinal tissues. Additionally, we detected abundant IL-7 transcripts in isolated LECs, suggesting that LECs produce IL-7, a heretofore unknown finding. Furthermore, GFP is expressed in a subpopulation of intestinal epithelial cells, and that expression was markedly upregulated in a dextran sulfate sodium-induced acute colitis model. Overall, IL-7-GFP knock-in mice serve as a unique and powerful tool to examine the identity and distribution of IL-7-expressing cells in vivo.


Linking knowledge with action when engagement is out of reach: three contextual features of effective public health communication.
Scholars and practitioners often promote direct engagement between policymakers, health workers and researchers as a strategy for overcoming barriers to utilizing scientific knowledge in health policy. However, in many settings public health officials rarely have opportunities to interact with researchers, which is a problem further exacerbated by the coronavirus disease 2019 pandemic. One prominent theory argues that policy actors will trust and utilize research findings when they perceive them to be salient, credible and legitimate. We draw on this theory to examine the conditions facilitating greater uptake of new knowledge among health officials when engagement is out of reach and they are instead exposed to new ideas through written mass communication. Using data from a survey experiment with about 260 health workers and administrators in Honduras, we find that messages from a technocratic sender based on statistical evidence improved perceptions of salience, credibility and legitimacy (SCL). Additionally, perceptions of SCL are three contextual features that operate as joint mediators between knowledge and action, and several individual characteristics also influence whether officials trust research findings enough to apply them when formulating and implementing health policies. This research can help inform the design of context-sensitive knowledge translation and exchange strategies to advance the goals of evidence-based public health, particularly in settings where direct engagement is difficult to achieve.


Dynamics and diversity of bacteria associated with the disease vectors Aedes aegypti and Aedes albopictus.
Aedes aegypti and Aedes albopictus develop in the same aquatic sites where they encounter microorganisms that influence their life history and capacity to transmit human arboviruses. Some bacteria such as Wolbachia are currently being considered for the control of Dengue, Chikungunya and Zika. Yet little is known about the dynamics and diversity of Aedes-associated bacteria, including larval habitat features that shape their tempo-spatial distribution. We applied large-scale 16S rRNA amplicon sequencing to 960 adults and larvae of both Ae. aegypti and Ae. albopictus mosquitoes from 59 sampling sites widely distributed across nine provinces of Panama. We find both species share a limited, yet highly variable core microbiota, reflecting high stochasticity within their oviposition habitats. Despite sharing a large proportion of microbiota, Ae. aegypti harbours higher bacterial diversity than Ae. albopictus, primarily due to rarer bacterial groups at the larval stage. We find significant differences between the bacterial communities of larvae and adult mosquitoes, and among samples from metal and ceramic containers. However, we find little support for geography, water temperature and pH as predictors of bacterial associates. We report a low incidence of natural Wolbachia infection for both Aedes and its geographical distribution. This baseline information provides a foundation for studies on the functions and interactions of Aedes-associated bacteria with consequences for bio-control within Panama.


Comparison of Intraoperative Computed Tomography Scan with Postoperative Magnetic Resonance Imaging for Determining Deep Brain Stimulation Electrode Coordinates.
Deep brain stimulation (DBS) is an effective therapy for a variety of refractory movement disorders. Accurate lead placement in the target nucleus is critical to ensure therapeutic effects and to minimize side effects, and intraoperative computed tomography (iCT) scan has been used to target and confirm lead position. The objective of this study is to compare the accuracy of determining the x, y, and z coordinates of final lead placement using iCT scan relative to postoperative magnetic resonance imaging (MRI). We conducted a retrospective study on 83 patients who underwent insertion of 145 DBS leads from 2015 to 2017 at a single institution. iCT scan was merged with the preoperative MRI to determine lead coordinates on both magnetic resonance and computed tomography images independently, and the absolute differences between the x, y, and z coordinates between the 2 scans along with the Euclidean vectors were calculated. The mean absolute differences ± standard error of the mean between iCT scan and postoperative MRI coordinates were as follows: x = 0.01 ± 0.09 mm (P = 0.89), y = 1.67 ± 0.14 mm (P < 0.001), and z = 2.75 ± 0.15 mm (P < 0.001). The average Euclidean vector difference was 3.21 ± 0.15 mm (P < 0.001). Significant differences exist between iCT scan and postoperative MRI DBS y and z lead coordinates, but not with x coordinates. Based on this series, iCT scan is more accurate when confirming x coordinates, and less accurate for confirming y and z coordinates during DBS operations.


Ligand effects on membrane lipids associated with sodium, potassium-activated adenosine triphosphatase: comparative spin probe studies with rat brain and heart enzyme preparations.
Physical properties of membrane lipids associated with rat or dog brain and heart Na+, K+-ATPase preparations were compared using an electron spin resonance probe, 5-doxyl stearate. The degree of acyl chain order of the membrane lipids was greater for brain enzyme than for heart enzyme preparations; membrane lipids in the rat heart enzyme preparations were the most disordered. In the absence of added ligands, membrane lipids did not appear to undergo a detectable temperature-dependent rearrangement or structural transition. An apparent transition was observed in the simultaneous presence of Na+, Mg2+, and ATP. These ligands increased lipid order at temperatures above the structural transition, but not below it. In the presence of the above ligands, K+ caused a marked decrease in the transition temperature in the rat brain enzyme preparations, but only a modest decrease in rat heart enzyme preparations. Arrhenius plots of rat brain and heart Na+, K+-ATPase activity revealed a break point roughly corresponding to respective membrane lipid transition temperatures observed in the presence of Na+, K+, Mg2+, and ATP. A low concentration of ouabain (1 microM) failed to affect either the lipid transition temperature estimated by the spin probe or the value of lipid order of the rat brain enzyme preparations observed in the presence of Na+, Mg2+, and ATP, but markedly reduced the effect of K+ to lower the transition temperature observed in the presence of the above ligands. A high concentration (100 microM) of ouabain which was needed to completely inhibit rat heart enzyme eliminated the thermally induced structural rearrangement observed in the presence of Na+, Mg2+, and ATP, apparently through a nonspecific lipid perturbation. These results indicate that differences in the physical properties of the membrane lipids per se are unlikely to account for the low affinity of rat heart Na+, K+-ATPase for ouabain and also suggest that the use of high concentrations of ouabain required to completely inhibit Na+, K+-ATPase activity may cause nonspecific changes in addition to inhibition of Na+, K+-ATPase or the sodium pump.


Modulation of repair of O6-methylguanine in parenchymal and nonparenchymal liver cells of rats treated with dimethylnitrosamine.
Previous studies have shown that chronic treatment of rats with dimethylnitrosamine (DMN) (2 mg/kg for 3 weeks) results in increased repair of O6-methylguanine (O6-mGua) in liver DNA. The experiments reported here try to determine if this increased repair is confined to one or more cell populations of the liver. Liver cells [parenchymal (PC) and nonparenchymal (NPC)] were separated by elutriation centrifugation at various times after the last administration of DMN. The in vivo alkylation studies show that at any time after a dose of [14C]DMN (2 mg/kg) the level of O6-mGua in PC cells of DMN pretreated rats was much lower than in the same cell population from control rats receiving only a single dose of DMN. In contrast, the pretreatment schedule resulted in no change in the levels of this DNA adduct in NPC cells. These results were confirmed by the determination of the levels of O6-methyldeoxyguanosine by radioimmunoassay in DNA from PC or NPC cells of rats similarly either pretreated for 3 weeks with DMN or receiving a single dose of DMN (2 mg/kg). The in vitro measurements of O6-mGua DNA alkyltransferase activity, using alkylated DNA as substrate, also show a higher activity of this repair enzyme in PC cells. The DMN pretreatment resulted in a 25-fold difference in O6-mGua DNA alkyltransferase activity between the two cell populations of the liver.


Hormonal induction of vascular tissue in tobacco pith in vitro.
The direct addition of indole-3-acetic acid into sterile-cultured stem pith sections of Nicotiana tabacum through inserted glass pipettes has induced cell division and differentiation where such activity would not normally occur. This evidence lends support to the theory that indole-3-acetic acid is the hormone that limits xylogenesis.


Utilization, attitudes, and experiences of Vietnam Era veterans with Veterans Administration health facilities: the American Legion experience.
A random sample of American Legion members in six states who had served in the Armed Forces during the Vietnam Era was conducted through a mailed questionnaire, in order to determine patterns of usage of Veterans Administration health facilities, as well as attitudes toward the VA and experiences at these facilities. Of the 6810 male respondents, 42.0% had served in Southeast Asia. These subjects were categorized according to their level of combat in South Vietnam. Thirty-six percent of those who had served in Southeast Asia had used VA health facilities, compared to only 18% of men who served elsewhere. Among Southeast Asia veterans, combat level was an important predictor of extent of usage of VA facilities for problems of both physical and mental health. Combat level was also associated with lack of basic and major medical insurance. While men with lower incomes tended to make greater use of VA mental health facilities, nearly one-fourth of mental health users had family incomes above $30,000. Despite their greater usage of the VA, men with higher combat levels expressed lower feelings of security about this agency, and rated its staff less helpful and of lower quality than did men who experienced lower levels of combat. On the other hand, higher combat veterans thought themselves better informed about VA services. Men who had gone to the VA for mental health assistance reported a disturbingly low frequency of having been asked basic questions that relate to possible diagnosis of post-traumatic stress disorder (PTSD), questions related to combat, which may be one of its etiologic factors, or other questions relating to their military history. Because combat level in Vietnam veterans is a major determinant of both attitudes toward and utilization of VA health facilities, the VA as well as other health agencies which deal with Vietnam veterans should be especially sensitive to this factor, and should take it into consideration when evaluating veterans' physical and mental health.


Integration host factor interacts with the DNA replication enhancer of filamentous phage f1.
We present data which show that the Escherichia coli integration host factor (IHF) is an activator of phage f1 DNA replication. Phage f1 poorly infects bacterial strains lacking IHF because IHF is required for efficient expression of F-pili, the receptor for f1 phage. However, when F- strains are transfected with f1 DNA the phage replicates in IHF mutants (himA, himD, or himA himD) at a rate of only 3% of that in wild-type bacteria. A plasmid dependent on the f1 replicon fails to transform IHF mutants. By gel retardation analysis, we show that IHF specifically binds to the origin of replication. DNase I "footprinting" experiments demonstrate that IHF binds to multiple sites within the replication enhancer sequence, a cis-acting, A + T-rich sequence that potentiates f1 DNA replication. Moreover, the effect of IHF mutation on f1 growth is suppressed by initiator protein (f1 gene II) mutations that restore efficient replication from origins that lack a functional replication enhancer sequence. This genetic evidence supports the conclusion that the replication enhancer sequence is the site of action of IHF.


Quercetin Prevents Oxidative Stress-Induced Injury of Periodontal Ligament Cells and Alveolar Bone Loss in Periodontitis.
Emerging evidence has indicated that oxidative stress (OS) contributes to periodontitis. Periodontal ligament cells (PDLCs) are important for the regeneration of periodontal tissue. Quercetin, which is extracted from fruits and vegetables, has strong antioxidant capabilities. However, whether and how quercetin affects oxidative damage in PDLCs during periodontitis remains unknown. The aim of this study was to assess the effects of quercetin on oxidative damage in PDLCs and alveolar bone loss in periodontitis and underlying mechanisms. The tissue block culture method was used to extract human PDLCs (hPDLCs). First, a cell counting kit 8 (CCK-8) assay was used to identify the optimal concentrations of hydrogen peroxide (H2O2) and quercetin. Subsequently, a 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) probe, RT-qPCR, Western blotting and other methods were used to explore the effects of quercetin on OS in hPDLCs and the underlying mechanism. Finally, quercetin was administered to mice with periodontitis through gavage, and the effect of quercetin on the level of OS and alveolar bone resorption in these mice was observed by immunofluorescence, microcomputed tomography (micro-CT), hematoxylin and eosin staining (H&E) staining and so on. Quercetin at 5 μM strongly activated NF-E2-related factor 2 (NRF2) signaling, alleviated oxidative damage and enhanced the antioxidant capacity of hPDLCs. In addition, quercetin reduced cellular senescence and protected the osteogenic ability of hPDLCs. Finally, quercetin activated NRF2 signaling in the periodontal ligaments, reduced the OS level of mice with periodontitis, and slowed the absorption of alveolar bone in vivo. Quercetin can increase the antioxidant capacity of PDLCs and reduce OS damage by activating the NRF2 signaling pathway, which alleviates alveolar bone loss in periodontitis.


The political and social barriers for contraception in pest birds: a case study of Ovocontrol (nicarbazin).
In a joint program, Innolytics and the U.S. Department of Agriculture/Animal and Plant Health Inspection Service National Wildlife Research Center collaborated in the development of nicarbazin as an avian contraceptive, initially for resident Canada geese and subsequently for feral pigeons. Unfortunately, the introduction of the original goose product in 2005 was a commercial failure. Political and social barriers effectively thwarted attempts to establish the new technology with any meaningful market success. Although the market adoption of the pigeon contraceptive has been less difficult, the product still encounters significant social and political obstacles and opposition. Given the focus on instant results and gratification, the introduction of contraceptive technology for birds has been challenging and broad market acceptance remains elusive. Nevertheless, especially for short-lived and rapidly reproducing species, customers continue to replace outdated or ineffective techniques with the safer and more effective contraceptive tool.


Signaling by a Conserved Quorum Sensing Pathway Contributes to Growth Ex Vivo and Oropharyngeal Colonization of Human Pathogen Group A Streptococcus.
Bacterial virulence factor production is a highly coordinated process. The temporal pattern of bacterial gene expression varies in different host anatomic sites to overcome niche-specific challenges. The human pathogen group A streptococcus (GAS) produces a potent secreted protease, SpeB, that is crucial for pathogenesis. Recently, we discovered that a quorum sensing pathway comprised of a leaderless short peptide, SpeB-inducing peptide (SIP), and a cytosolic global regulator, RopB, controls speB expression in concert with bacterial population density. The SIP signaling pathway is active in vivo and contributes significantly to GAS invasive infections. In the current study, we investigated the role of the SIP signaling pathway in GAS-host interactions during oropharyngeal colonization. The SIP signaling pathway is functional during growth ex vivo in human saliva. SIP-mediated speB expression plays a crucial role in GAS colonization of the mouse oropharynx. GAS employs a distinct pattern of SpeB production during growth ex vivo in saliva that includes a transient burst of speB expression during early stages of growth coupled with sustained levels of secreted SpeB protein. SpeB production aids GAS survival by degrading LL37, an abundant human antimicrobial peptide. We found that SIP signaling occurs during growth in human blood ex vivo. Moreover, the SIP signaling pathway is critical for GAS survival in blood. SIP-dependent speB regulation is functional in strains of diverse emm types, indicating that SIP signaling is a conserved virulence regulatory mechanism. Our discoveries have implications for future translational studies.


[Functional coupling of creatine phosphokinase and adenylate kinase with adenine nucleotide translocase and its role in regulation of heart mitochondrial respiration].
The respiration of rabbit heart mitochondria in the presence of ATP is stimulated by ADP, AMP, creatine and glucose plus hexokinase. The values of V for mitochondrial phosphorylating respiration in the presence of corresponding stimulators are equal to 491 +/- 34, 460 +/- 12, 480 +/- 45 and 463 +/- 72 natoms O2 X min-1 X mg-1 of protein, 37 degrees C. The half-maximal stimulation of respiration is observed at 35 microM AMP, 60 microM ADP and 10 mM creatine in the absence of creatine phosphate. In the presence of creatine phosphate the maximal stimulation of heart mitochondrial respiration is achieved under a combined action of creatine and AMP. The inhibition type of mitochondrial respiration by palmitoyl-CoA depends on the nature of stimulators used. Thus, with ADP or glucose plus hexokinase the inhibition is competitive, while with AMP and creatine an uncompetitive and non-competitive inhibition was observed, respectively. The experimental results are indicative of functional coupling of heart mitochondrial adenylate kinase and creatine phosphokinase with ATP-ADP-translocase. It is assumed that creatine and AMP act as physiological regulators of heart mitochondrial respiration ("feed-back" signals from cytoplasm to mitochondria).


Proteomic interrogation of venom delivery in marine cone snails: novel insights into the role of the venom bulb.
Cone snails of the genus Conus are predatory marine gastropods mainly found in the shallow waters of the tropics and warm temperate seas. To prey on other marine organisms including fish, cone snails have evolved complex venoms synthesized and delivered by a highly sophisticated venom apparatus. Upon prey discovery, the venom is perfused through a harpoon-like radula tooth and rapidly injected into the prey to cause paralysis. While the venom components of cone snails have been intensively characterized, the mechanism of venom translocation and loading prior to and during injection remains elusive. The involvement of the venom bulb, a muscular dilation of the venom gland has been suggested, however evidence is sparse. Here, we use a combination of proteomics, molecular biology, and morphological examination to elucidate the potential role of the venom bulb in venom translocation and delivery. Analysis of the venom bulb proteome clearly demonstrated a function of this organ in muscular movement and, more interestingly, in burst muscle contraction. Morphological examination revealed high structural similarities to the mantle muscle of squids, animals known for their rapid escape response. We sequenced and further characterized arginine kinase, a key protein of rapid muscular movement in invertebrates and show high concentrations of this enzyme in the bulb when compared to the venom gland and the foot muscle. Proteins characteristic for venom biosynthesis were low in abundance. On the basis of our findings, we suggest that the bulb of cone snails is a highly specialized organ of venom translocation. Delivery of venom is driven by burst contractions of the bulb rapidly forcing the venom through the radula tooth into the prey.


Proinflammatory mediator-induced reversal of CD4+,CD25+ regulatory T cell-mediated suppression in rheumatoid arthritis.
We previously demonstrated that CD4+,CD25+ regulatory T (Treg) cells are present in increased numbers in the synovial fluid (SF) of rheumatoid arthritis (RA) patients and display enhanced suppressive activity as compared with their peripheral blood (PB) counterparts. Despite the presence of these immunoregulatory cells in RA, chronic inflammation persists. The purpose of the present study was to investigate whether particular proinflammatory mediators that are associated with RA could abrogate CD4+,CD25+ Treg-mediated suppression. Monocyte phenotype was determined by flow cytometry and cytokine levels by enzyme-linked immunosorbent assay. Magnetically sorted CD4+,CD25- and CD4+,CD25+ T cells derived from the PB and SF obtained from RA patients were stimulated alone or in coculture with anti-CD3 monoclonal antibody (mAb) and autologous antigen-presenting cells, in the absence or presence of anti-CD28 mAb or the proinflammatory cytokines interleukin-6 (IL-6), tumor necrosis factor alpha (TNFalpha), or IL-7. Monocytes from the SF of RA patients displayed increased expression of HLA class II molecules, CD80, CD86, and CD40 as compared with PB-derived monocytes, indicating their activated status. Mimicking this increased costimulatory potential, addition of anti-CD28 mAb to cocultures of CD4+,CD25- and CD4+,CD25+ T cells resulted in reduced CD4+,CD25+ Treg-mediated suppression in both PB and SF. Furthermore, IL-7 and, to a limited extent, TNFalpha, both of which are produced by activated monocytes and were detected in SF, abrogated the CD4+,CD25+ Treg-mediated suppression. In contrast, IL-6 did not influence Treg-mediated suppression. Our findings suggest that the interaction of CD4+,CD25+ Treg cells with activated monocytes in the joint might lead to diminished suppressive activity of CD4+,CD25+ Treg cells in vivo, thus contributing to the chronic inflammation in RA.


Ventricular arrhythmias late after myocardial infarction are related to hypomagnesemia and magnesium loss: preliminary trial of corrective therapy.
It has been well established that in acute myocardial infarction (MI) many patients display low serum magnesium (Mg). This is associated with complex ventricular arrhythmias. The question arises whether predischarge arrhythmias occurring late after MI might also be related to Mg imbalance. In 118 patients subjected to heart rhythm 24 h Holter monitoring in the second or third week after MI, we investigated (1) the relationship between serum Mg, urinary Mg loss, and ventricular arrhythmias, and (2) the effect of Mg supplementation on heart rhythm disturbances. In patients with undisturbed rhythm or monomorphic ventricular ectopic beats (VEB) (Lown 0-2; n = 84), mean serum Mg level (mg% +/- SD) was 1.83 +/- 0.21, whereas in patients with multifocal VEB, pairs, or nonsustained ventricular tachycardia (VT) (Lown 3-4; n = 34) serum Mg was decreased to 1.68 +/- 0.27 (p < 0.01). Serum Mg normal range in our laboratory is 1.7-2.6 mg%. The lowest serum Mg reaching 1.55 +/- 0.27 was found in nonsustained VT (Lown 4 b) subgroup (n = 14). Urinary Mg loss measured in 81 patients was more pronounced in those with Lown 3-4 arrhythmias (n = 26) than with Lown 0-2 (n = 55). The daily values were 73 +/- 22 and 54.4 +/- 26 mg, respectively (p < 0.001). Thirteen patients with complex arrhythmias and low serum Mg received Mg supplementation (MgSO4, 8 g in 500 ml 5% glucose intravenously during 24 h). This resulted in restoration of almost undisturbed rhythm in 10 subjects.(ABSTRACT TRUNCATED AT 250 WORDS)


[The efficacy and safety of cefixime and amoxicillin/clavulanate in the treatment of asymptomatic bacteriuria in pregnant women: a randomized, prospective, multicenter study].
The study was aimed to the evaluation of efficacy and safety of cefixime and amoxicillin/clavulanate in the treatment of asymptomatic bacteriuria in pregnant women. A prospective, multicenter, randomized study that included 112 pregnant women with asymptomatic bacteriuria was performed. 58 women were randomized in group 1 (cefixime [suprax solutab] 400 mg 1 time a day, 7 days), 54 women were included in group 2 (amoxicillin/clavulanate [amoksiklav] 625 mg 3 times a day, 7 days). The average age of the patients in group 1 was 25.2 +/- 6.6; in group 2--26.6 +/- 5.8 years. Physical examination, evaluation of complaints, collection of data on adverse reactions, and bacteriological analysis of urine were performed after enrollment in the study at visit 2 (day 10 +/- 1) and 3 (day 35 +/- 2). Comparable effectiveness of cefixime and amoxicillin/clavulanate in the treatment of asymptomatic bacteriuria in pregnant women was found. Eradication of the pathogen and sustained bacteriological response were observed in 94.8 and 92.7% of women treated with cefixime, and in 98.2 and 92.5% of women treated with amoxicillin/clavulanate, respectively (P > 0.05). At the same time, the use of amoxicillin/clavulanate compared with cefixime significantly higher was followed by the development of adverse reactions (13% and 1.7; respectively; P = 0.02). Seven-day courses of cefixime at a dose 400 mg 1 time a day and amoxicillin/clavulanate at a dose of 625 mg 3 times a day are high-effective treatment regimens for asymptomatic bacteriuria in pregnant women in Russia. The use of amoxicillin/clavulanate is significantly more often accompanied by the development of adverse reactions compared with cefixime.


Differentiation of population of peripheral blood lymphocytes into cells bearing sheep erythrocyte receptors in vitro by human thymic extract.
A small population of human marrow cells has been shown to be differentiated in vitro by thymic extract into cells bearing T-lymphocyte (thymus-derived lymphocyte) characteristics. By a similar method, the differentiation of human peripheral blood lymphocytes has been studied. A discontinuous gradient of bovine serum albumin was used to isolate lymphocytes into four layers and cells from layers I and III demonstrated the greatest potential for differentiation by human thymic extract. Appearance of T-lymphocyte characteristics was recognized by the spontaneous E-rosette technique with sheep erythrocytes. Ability of human marrow cells to be differentiated under the influence of human thymic extract was abolished by specific inhibitors of nucleic acid synthesis, however, had no inhibitory effect on the maturation of peripheral blood lymphocytes during a 2 hr incubation with human thymic extract but puromycin, an inhibitor of protein synthesis, abolished this differentiative step in cells of layer I. It is suggested from these studies that many of the cells in peripheral blood that are differentiable by thymic extract are at a stage of maturation more advanced than those in human marrow that are also differentiable by thymic extract.


A new in vitro study demonstrates that electronic cigarettes promote cellular carcinogenic characteristics.
Electronic cigarettes have been used as a new form of cigarettes, especially among young people, but the impact of these effects on mouth cancer is unknown. Therefore, there is a need for studies to evaluate their impact on health and oral mucosa. In this way, this study evaluates the risk of electronic cigarette liquid on in vitro cells of a panel: normal oral epithelium cell lines (NOE and HMK), oral squamous cell carcinoma human cell lines (CAL27 and HSC3), and a mouse oral cancer cell line (AT84). It was demonstrated that electronic cigarettes promote proliferation and anchorage-independent growth and induces morphological changes associated with enhanced motility and invasive phenotypes. Also, it was observed that the enhanced invasive migratory activity associated with the loss of epithelial markers, such as E-cadherin, and the acquisition of mesenchymal markers, strongly suggest that epithelial cells are undergoing to an aggressive phenotype within the framework of epithelial-mesenchymal transition. In conclusion, the liquid can promote carcinogenesis, in addition to promoting an aggressive phenotype in pre-existing lesions.


Developmental expression and tissue distribution of Phex protein: effect of the Hyp mutation and relationship to bone markers.
Mutations in PHEX, a phosphate-regulating gene with homology to endopeptidases on the X chromosome, are responsible for X-linked hypophosphatemia (XLH). The murine Hyp homologue has the phenotypic features of XLH and harbors a large deletion in the 3' region of the Phex gene. We characterized the developmental expression and tissue distribution of Phex protein, using a monoclonal antibody against human PHEX, examined the effect of the Hyp mutation on Phex expression, and compared neprilysin (NEP), osteocalcin, and parathyroid hormone/parathyroid hormone-related protein (PTH/PTHrP) receptor gene expression in bone of normal and Hyp mice. Phex encodes a 100- to 105-kDa glycoprotein, which is present in bones and teeth of normal mice but not Hyp animals. These results were confirmed by in situ hybridization (ISH) and ribonuclease protection assay. Phex protein expression in femur and calvaria decreases with age, suggesting a correlation between Phex expression and bone formation. Immunohistochemical studies detected Phex protein in osteoblasts, osteocytes, and odontoblasts, but not in osteoblast precursors. In contrast to Phex, the abundance of NEP messenger RNA (mRNA) and protein is not significantly altered in Hyp bone. Similarly, osteocalcin and PTH/PTHrP receptor gene expression are not compromised in bone of Hyp mice. Our results are consistent with the hypothesis that loss of Phex function affects the mineralizing activity of osteoblasts rather than their differentiation.


[Specific features of materials for initial pulpitis treatment].
The aim of the current study was to assess antibacterial and cytotoxic properties of Biodentine (Septodont), Rootdent (TehnoDent) and adhesive Futurabond НР (Voco). Two lines of experiments were carried out using cements water solutions and firm tablet-like samples (made by means of special pattern). Citotoxic activity was tested on NCTC L929 mice line fibroblasts culture. All the examined materials showed antibacterial activity against E. coli, S. aureus, C. albiсans, St. faecalis, mostly evident in Futurabond and the poorest in Biodentine samples. As for cytotoxic properties, Biodentine proved not to suppress metabolic activity stimulating odontotropic impact. The results confirm the analyzed materials to be a useful tool for deep caries lesions and initial pulpitis treatment.


Neutral endopeptidase regulates C-type natriuretic peptide metabolism but does not potentiate its bioactivity in vivo.
C-type natriuretic peptide (CNP) is a newly described 22-amino acid peptide of endothelial and renal cell origin with selective cardiovascular actions. Recent in vitro studies have reported that CNP is the most susceptible of all natriuretic peptides to enzymatic degradation by neutral endopeptidase 24.11 (NEP). The present study was undertaken to define the role of NEP in total and regional CNP metabolism and the modulatory actions of NEP inhibition on the biological actions of CNP. CNP (10 ng x kg(-1) x min(-1)) followed by candoxatrilat (240 microg x kg(-1) bolus and 8 microg x kg(-1) x min(-1)), a potent and selective NEP inhibitor, was administered intravenously to a group of anesthetized mongrel dogs (group 1) to permit calculation of total metabolic clearance rate (MCR); results were compared with those in a group receiving vehicle infusion followed by candoxatrilat (group 2; both groups, n=7). NEP inhibition increased circulating CNP achieved by exogenous infusion and reduced total MCR in group 1. The regional CNP MCRs increased after CNP administration. While the pulmonary MCR did not change during concomitant candoxatrilat infusion, renal MCR was suppressed. Hemodynamic changes were not different between groups. A mild natriuretic and diuretic effect in association with an increase in circulating and urinary ANP levels was not different between groups. Urinary CNP excretion did not change with CNP infusion but markedly increased after NEP inhibition. We conclude that (1) circulating CNP achieved by exogenous CNP infusion is regulated by NEP in vivo, (2) regional MCRs are heterogeneous with NEP inhibition, (3) NEP inhibition does not potentiate acute cardiovascular actions of CNP, and (4) a mild natriuretic and diuretic effect observed with CNP and NEP inhibition may be ANP dependent.


Development and validation of a sensitive LC-MS-MS method to quantify psilocin in authentic oral fluid samples.
Psilocin is an active substance and a dephosphorylated product of psilocybin formed after the ingestion of mushrooms. The low stability caused by the quick oxidation of this analyte requires sensitive methods for its determination in biological matrices. In this work, we described the development, optimization and validation of a method for the quantification of psilocin in authentic oral fluid samples by liquid chromatography-tandem mass spectrometry. Liquid-liquid extraction was performed using 100 µL of oral fluid samples collected with a Quantisal™ device and t-butyl methyl ether as the extraction solvent. The method showed acceptable performance, with limits of detection and quantification of 0.05 ng/mL, and the calibration model was achieved between 0.05 and 10 ng/mL. Bias and imprecision results were below -14.2% and 10.7%, respectively. Ionization suppression/enhancement was lower than -30.5%, and recovery was >54.5%. Dilution integrity bias was <14.4%. No endogenous and exogenous interferences were observed upon analyzing oral fluid from 10 different sources and 56 pharmaceuticals and drugs of abuse, respectively. No carryover was observed at 10 ng/mL. Psilocin was stable in oral fluid at -20°C, 4°C and 24°C up to 24, 72 and 24 h, respectively, with variations <17.7%. The analyte was not stable after three freeze/thaw cycles, with variations between -73% and -60%. This suggests the instability of psilocin in oral fluid samples, which requires timely analysis, as soon as possible after the collection. The analyte remained stable in processed samples in an autosampler (at 10°C) for up to 18 h. The method was successfully applied for the quantification of five authentic samples collected from volunteers attending parties and electronic music festivals. Psilocin concentrations ranged from 0.08 to 36.4 ng/mL. This is the first work to report psilocin concentrations in authentic oral fluid samples.


Introductory tail-flick of the Jacky dragon visual display: signal efficacy depends upon duration.
Many animal signals have introductory components that alert receivers. Examples from the acoustic and visual domains show that this effect is often achieved with high intensity, a simple structure and a short duration. Quantitative analyses of the Jacky dragon Amphibolurus muricatus visual display reveal a different design: the introductory tail-flick has a lower velocity than subsequent components of the signal, but a longer duration. Here, using a series of video playback experiments with a digitally animated tail, we identify the properties responsible for signal efficacy. We began by validating the use of the computer-generated tail, comparing the responses to digital video footage of a lizard tail-flick with those to a precisely matched 3-D animation (Experiment 1). We then examined the effects of variation in stimulus speed, acceleration, duration and period by expanding and compressing the time scale of the sequence (Experiment 2). The results identified several variables that might mediate recognition. Two follow-up studies assessed the importance of tail-flick amplitude (Experiment 3), movement speed and signal duration (Experiment 4). Lizard responses to this array of stimuli reveal that duration is the most important characteristic of the tail-flick, and that intermittent signalling has the same effect as continuous movement. We suggest that signal design may reflect a trade-off between efficacy and cost.


Carotid stenting and "extarterectomy" in the management of head and neck cancer involving the internal carotid artery: technical case report.
Head and neck cancer that invades the internal carotid artery (ICA) represents a significant management challenge. We describe a novel technique that allows for aggressive tumor removal without disrupting blood flow through the affected ICA. A 62-year-old man was referred to our institution for management of a neck malignancy involving the ICA. Cerebral angiography suggested that there was good collateral flow from the opposite hemisphere, but the patient reported visual loss in the ipsilateral eye during balloon test occlusion of the ICA. A self-expanding stent was deployed in the ICA; it spanned the entire length of the artery involved by tumor. One month later, the patient underwent tumor resection. During surgery, a long ICA arteriotomy was performed directly down to the mesh of the stent. A neoendothelium had formed within the stent, which prevented arterial bleeding. The carotid wall was dissected from the stent without difficulty and removed en bloc with the surrounding tumor. The exposed stent was wrapped circumferentially with a synthetic patch material. The patient tolerated the procedure well, and postoperative angiography demonstrated normal filling of the ICA. We describe a novel approach to a patient with head and neck cancer involving the cervical ICA. Preliminary stenting, which allows time for endothelialization before surgery, may permit aggressive tumor resection without interrupting flow through the ICA. This technique obviates the need for complicated carotid reconstruction procedures and avoids the risk of delayed ischemia from carotid sacrifice.


The in vitro effect of different PRP concentrations on osteoblasts and fibroblasts.
The aim of this study was to assess the biological rationale for the use of platelet-rich plasma (PRP) by evaluating the effect of different concentrations of PRP on osteoblasts (OB) and fibroblasts (FB) function in vitro. PRP was obtained from volunteer donors using standard protocols. Primary human cultures of oral FBs and OBs were exposed to both activated and non-activated plasma as well as various concentrations of PRP (2.5 x, 3.5 x and max (4.2-5.5 x)). Cell proliferation was evaluated after 24 and 72 h using an MTT proliferation assay. Production of osteocalcin (OCN), osteoprotegerin (OPG) and transforming growth factor beta1 (TGF-beta1) was evaluated in OB after 24 and 72 h. Statistical analysis was performed using one-way ANOVA. PRP-stimulated cell proliferation in both OBs and FBs. The effect of different PRP concentrations on cell proliferation was most notable at 72 h. The maximum effect was achieved with a concentration of 2.5 x, with higher concentrations resulting in a reduction of cell proliferation. Upregulation of OCN levels and downregulation of OPG levels were noted with increasing PRP concentrations at both 24 and 72 h. TGF-beta1 levels were stimulated by increasing concentrations of PRP, with the increased levels being maintained at 72 h. PRP preparations exert a dose-specific effect on oral FBs and OBs. Optimal results were observed at a platelet concentration of 2.5 x, which was approximately half of the maximal concentrate that could be obtained. Increased concentrations resulted in a reduction in proliferation and a suboptimal effect on OB function. Hence, different PRP concentrations may have an impact on the results that can be obtained in vivo.


Tri- and hexaferrocenyl-substituted subphthalocyanines in the quest for the optimum electron donor-acceptor distances.
Three and six ferrocenyl subunits have been attached to the periphery of subphthalocyanines (SubPcs). Unlike axially coordinated ferrocenes, peripherally-bonded ferrocenes have an impact on the electronic features of SubPcs, which show a 44 to 70 nm red-shift of their Q-bands. The unusually deep and narrow ferrocenyl-SubPc is able to host C60, giving rise to atypical SubPc•C60 cocrystallates, through a combination of concave-convex and convex-convex π-π interactions.


Protective effects of rosmarinic acid against hydrogen peroxide‑induced cellular senescence and the inflammatory response in normal human dermal fibroblasts.
Hydrogen peroxide (H2O2) is a reactive oxygen species (ROS) that induces numerous cellular events, including cellular senescence and inflammatory responses. Therefore, the aim of this study was to investigate the protective effect of Rosmarinic acid (RA) in H2O2‑induced oxidative stress in normal human dermal fibroblasts (NHDFs). Cytotoxicity assays were performed using a water‑soluble tetrazolium salt, and senescence‑associated β‑galactosidase activity was determined to investigate the proportion of senescent cells. Antioxidant capacities were evaluated via H2O2‑scavenging activity, reverse transcription‑quantitative polymerase chain reaction, NRF2 luciferase reporter gene activity and intracellular ROS scavenging assays. Cytokine‑coded gene expression analysis and nuclear factor‑κB luciferase activity were determined to verify the anti‑inflammatory effect of RA. As a result, the present study demonstrated that rosmarinic acid inhibited H2O2‑induced oxidative stress and inflammatory responses in normal human dermal fibroblasts. Initially, the doses of RA that exerted minimal cytotoxic effects in NHDFs were determined using a cytotoxicity assay. Subsequently, pretreatment with the appropriate doses of RA significantly reversed the H2O2‑induced decrease in NHDF cell viability and decreased cellular senescence of NHDFs. In addition, RA inhibited H2O2‑induced ROS production in NHDFs, as determined by a ROS scavenging assay. The protective effects of RA were mediated by the inhibition of nuclear factor erythroid‑derived 2‑like 2, a transcription factor that functions as a key regulator of redox sensitivity. Furthermore, RA suppressed H2O2‑induced inflammation in NHDFs and significantly rescued H2O2‑induced downregulation of sirtuin 1. RA also inhibited nuclear factor (NF)‑κB transcriptional activity and the expression of NF‑κB target genes, including tumor necrosis factor‑α and interleukin‑6, in H2O2‑exposed NHDFs. Taken together, these data indicate that RA inhibits H2O2‑induced cellular damage in NHDFs.


The diagnostic significance of enzyme linked immuno-sorbent assay for herpes simplex, varicella zoster and cytomegalovirus retinitis.
To evaluate the diagnostic usefulness of enzyme linked immuno-sorbent assay (ELISA) in single serum samples to associate herpes simplex virus (HSV), varicella zoster virus (VZV) or cytomegalovirus (CMV) with viral retinitis as against polymerase chain reaction (PCR) on intraocular specimens. It was also designed to study the seroprevalence in normal healthy individuals, and the genomic prevalence of HSV, VZV and CMV in patients without an active viral inflammatory process. PCR for the detection of HSV, VZV and CMV genomes was done on 33 and 90 intraocular fluids from viral retinal patients and non-viral controls respectively. ELISA was done on 30 and 100 serum samples from viral retinitis patients and normal healthy controls respectively. PCR did not detect HSV, VZV and CMV genomes except one, in which VZV-DNA was detected. ELISA showed prevalence rates of 28%, 83% and 90% for antibodies against HSV, VZV and CMV respectively in the normal population. In the 30 viral retinitis patients, PCR detected HSV-DNA in 2 (6.7%), VZV-DNA in 7 (23.3%) and CMV-DNA in 6 (20.0%) patients, while ELISA detected antibodies against HSV, VZV and CMV in 13 (43.3%), 24 (80.0%) and 23 (76.7%) patients respectively. ELISA was of value in indirect diagnosis only in 6 (20.0%) as compared to 15 (50.0%) of 30 patients by PCR, this difference was statistically significant (McNemar test, P value = 0.005). Serology by ELISA is no longer a useful diagnostic tool to associate HSV, VZV and CMV viruses with viral retinitis.


Human amniotic fluid stem cells seeded in fibroin scaffold produce in vivo mineralized matrix.
This study investigated the potential of amniotic fluid stem cells (AFSCs) to synthesize mineralized extracellular matrix (ECM) within different porous scaffolds of collagen, poly-D,L-lactic acid (PDLLA), and silk fibroin. The AFSCs were initially differentiated by using an osteogenic medium in two-dimensional culture, and expression of specific bone proteins and the physiologic mineral production by the AFSCs were analyzed. In particular, during differentiation process, AFSCs expressed proteins like Runt-related transcription factor 2 (Runx2), Osterix, Osteopontin, and Osteocalcin with a sequential expression, analogous to those occurring during osteoblast differentiation, and produced extracellular calcium stores. AFSCs were then cultured on three-dimensional (3D) scaffolds and evaluated for their ability to differentiate into osteoblastic cells in vivo. Stem cells were cultured in vitro for 1 week in collagen, fibroin, and PDLLA scaffolds. The effect of predifferentiation of the stem cells in scaffolds on the subsequent bone formation in vivo was determined in a rat subcutaneous model. With the addition of a third dimension, osteogenic differentiation and mineralized ECM production by AFSCs were significantly higher. This study demonstrated the strong potential of AFSCs to produce 3D mineralized bioengineered constructs in vivo and suggests that fibroin may be an effective scaffold material for functional repair of critical size bone defects.


Decreased reaction time variability is associated with greater cardiovascular responses to acute stress.
Cardiovascular (CV) responses to mental stress are prospectively associated with poor CV outcomes. The association between CV responses to mental stress and reaction times (RTs) in aging individuals may be important but warrants further investigation. The present study assessed RTs to examine associations with CV responses to mental stress in healthy, older individuals using robust regression techniques. Participants were 262 men and women (mean age = 63.3 ± 5.5 years) from the Whitehall II cohort who completed a RT task (Stroop) and underwent acute mental stress (mirror tracing) to elicit CV responses. Blood pressure, heart rate, and heart rate variability were measured at baseline, during acute stress, and through a 75-min recovery. RT measures were generated from an ex-Gaussian distribution that yielded three predictors: mu-RT, sigma-RT, and tau-RT, the mean, standard deviation, and mean of the exponential component of the normal distribution, respectively. Decreased intraindividual RT variability was marginally associated with greater systolic (B = -.009, SE = .005, p = .09) and diastolic (B = -.004, SE = .002, p = .08) blood pressure reactivity. Decreased intraindividual RT variability was associated with impaired systolic blood pressure recovery (B = -.007, SE = .003, p = .03) and impaired vagal tone (B = -.0047, SE = .0024, p = .045). Study findings offer tentative support for an association between RTs and CV responses. Despite small effect sizes and associations not consistent across predictors, these data may point to a link between intrinsic neuronal plasticity and CV responses.


Efficacy and safety of new oral anticoagulants for extended treatment of venous thromboembolism: systematic review and meta-analyses of randomized controlled trials.
Currently available anticoagulants have limitations for long term treatment of venous thromboembolism (VTE). A meta-analysis was performed to evaluate the efficacy and safety of new oral anticoagulants (NOACs) for extended treatment of VTE. PubMed, Cochrane Library, EMBASE, Web of Science and CINAHL databases were searched from January 01, 2001 through February 28, 2013. Randomized controlled trials (RCTs) comparing NOACs (apixaban, rivaroxaban and dabigatran) with placebo or warfarin for extended treatment of VTE were selected. Primary efficacy outcome was recurrent VTE or VTE related death, and primary safety outcome was major bleeding. We used random-effects models. Four RCTs included 7,877 participants. NOACs significantly lowered the risk of recurrent VTE or VTE-related death compared to placebo/warfarin (odds ratio [OR] 0.25, 95 % confidence interval [CI] 0.07 to 0.86; number needed to treat [NNT] = 30). All-cause mortality was significantly lower in NOACs group compared to placebo (OR 0.38, 95 % CI 0.18 to 0.80). Risk of major bleeding was not different with NOACs compared to placebo/warfarin (OR 0.88, 95 % CI 0.27 to 2.91). However, NOACs caused significantly higher rate of major or clinically relevant bleeding compared to placebo (OR 2.69, 95 % CI 1.25 to 5.77; number needed to harm [NNH] = 39). All three NOACs (apixaban, rivaroxaban and dabigatran) individually significantly reduced recurrent VTE or VTE-related death compared to placebo. Major or clinically relevant bleeding was higher with dabigatran and rivaroxaban but not with apixaban. NOACs are effective for the extended treatment of venous thromboembolism and may reduce the risk of all-cause mortality. Dabigatran and rivaroxaban may cause more major or clinically relevant bleeding.


Separation and characterization of respirable amphibole fibers from Libby, Montana.
The vermiculite mine in Libby, Montana, was in operation for over 70 yr and was contaminated with asbestos-like amphibole fibers. The mining, processing, and shipping of this vermiculite led to significant fiber inhalation exposure throughout the community, and residents of Libby have developed numerous pulmonary diseases such as lung cancer and mesothelioma. The present study describes the separation of Libby 6-mix into respirable and nonrespirable size fractions by means of aqueous elutriation. The elutriator, designed to separate fibers with aerodynamic diameters smaller than 2.5 microm (respirable) from larger fibers, used an upward flow rate of 3.4 x 10(- 4) cm s(-1). The resultant respirable fraction constituted only 13% of the raw Libby 6-mix mass, and less than 2% of the fibers in the elutriated fraction had aerodynamic diameters exceeding 2.5 microm. Surface area of the elutriated fibers was 5.3 m(- 2) g(-1), compared to 0.53 m(-2) g(-1) for the raw fibers. There were no detectable differences in chemical composition between the larger and smaller fibers. Such harvesting of respirable fractions will allow toxicological studies to be conducted within a controlled laboratory setting, utilizing fiber sizes that may more accurately simulate historical exposure of Libby residents' lungs. Importantly, this work describes a method that allows the use of material enriched in more uniform respirable material than raw Libby 6-mix, making comparisons with other known fiber preparations more valid on a mass basis.


Use of glucosamine and chondroitin in relation to mortality.
Glucosamine and chondroitin are products commonly used by older adults in the US and Europe. There is limited evidence that they have anti-inflammatory properties, which could provide risk reduction of several diseases. However, data on their long-term health effects is lacking. To evaluate whether use of glucosamine and chondroitin are associated with cause-specific and total mortality. Participants (n = 77,510) were members of a cohort study of Washington State (US) residents aged 50-76 years who entered the cohort in 2000-2002 by completing a baseline questionnaire that included questions on glucosamine and chondroitin use. Participants were followed for mortality through 2008 (n = 5,362 deaths). Hazard ratios (HR) for death adjusted for multiple covariates were estimated using Cox models. Current (baseline) glucosamine and chondroitin use were associated with a decreased risk of total mortality compared to never use. The adjusted HR associated with current use of glucosamine (with or without chondroitin) was 0.82 (95 % CI 0.75-0.90) and 0.86 (95 % CI 0.78-0.96) for chondroitin (included in two-thirds of glucosamine supplements). Current use of glucosamine was associated with a significant decreased risk of death from cancer (HR 0.87 95 % CI 0.76-0.98) and with a large risk reduction for death from respiratory diseases (HR 0.59 95 % CI 0.41-0.83). Use of glucosamine with or without chondroitin was associated with reduced total mortality and with reductions of several broad causes of death. Although bias cannot be ruled out, these results suggest that glucosamine may provide some mortality benefit.


β-caryophyllene reduces atherogenic index and coronary risk index in hypercholesterolemic rats: The involvement of cardiac oxidative damage.
Hypercholesterolemia and cardiac oxidative damage have been implicated in the pathophysiology of atherosclerosis. In earlier studies, treatment using natural phytocannabinoid β-caryophyllene caused a hypolipemic effect, as well as ameliorated hepatic oxidative damage. In this regard, the present study aimed to investigate whether β-caryophyllene treatment was able to protect the cardiac tissue against hypercholesterolemic atherosclerosis, and to investigate the involvement of antioxidant mechanisms in this effect using a model of hypercholesterolemia induced by Triton WR-1339. The hypercholesterolemic animals exhibited a significant increase in the cholesterol (TC), triglycerides (TG) and free fatty acids (FFA) contents in cardiac tissue, as well as showed significant increase on atherogenic index (AI) and coronary risk index (CRI). Moreover, cardiac levels of reactive oxygen species (ROS) and thiobarbituric acid reactive substances (TBARS) were increased in hypercholesterolemic animals, while the glutathione reduced (GSH) levels and glutathione peroxidase (GPx) activity were decreased. β-caryophyllene treatment reduced the cardiac lipid content, as well as decreased the AI and CRI. Moreover, the treatment was able to improve the antioxidant/oxidant status in heart tissue of hypercholesterolemic animals. In summary, β-caryophyllene possesses antioxidant properties, preventing lipidic oxidative damage and ameliorating the GPx activity, an important enzyme linked to the prevention of atherosclerosis. Thus, this compound can be used as an attempt to prevent or reduce atherosclerosis in hypercholesterolemic rats.


Effects of centrally acting ACE inhibitors on the rate of cognitive decline in dementia.
There is growing evidence that antihypertensive agents, particularly centrally acting ACE inhibitors (CACE-Is), which cross the blood-brain barrier, are associated with a reduced rate of cognitive decline. Given this, we compared the rates of cognitive decline in clinic patients with dementia receiving CACE-Is (CACE-I) with those not currently treated with CACE-Is (NoCACE-I), and with those who started CACE-Is, during their first 6 months of treatment (NewCACE-I). Observational case-control study. 2 university hospital memory clinics. 817 patients diagnosed with Alzheimer's disease, vascular or mixed dementia. Of these, 361 with valid cognitive scores were included for analysis, 85 CACE-I and 276 NoCACE-I. Patients were included if the baseline and end-point (standardised at 6 months apart) Standardised Mini-Mental State Examination (SMMSE) or Quick Mild Cognitive Impairment (Qmci) scores were available. Patients with comorbid depression or other dementia subtypes were excluded. The average 6-month rates of change in scores were compared between CACE-I, NoCACE-I and NewCACE-I patients. When the rate of decline was compared between groups, there was a significant difference in the median, 6-month rate of decline in Qmci scores between CACE-I (1.8 points) and NoCACE-I (2.1 points) patients (p=0.049), with similar, non-significant changes in SMMSE. Median SMMSE scores improved by 1.2 points in the first 6 months of CACE treatment (NewCACE-I), compared to a 0.8 point decline for the CACE-I (p=0.003) group and a 1 point decline for the NoCACE-I (p=0.001) group over the same period. Multivariate analysis, controlling for baseline characteristics, showed significant differences in the rates of decline, in SMMSE, between the three groups, p=0.002. Cognitive scores may improve in the first 6 months after CACE-I treatment and use of CACE-Is is associated with a reduced rate of cognitive decline in patients with dementia.


Genotyping of Burkholderia mallei from an outbreak of glanders in Bahrain suggests multiple introduction events.
Glanders, caused by the gram-negative bacterium Burkholderia mallei, is a highly infectious zoonotic disease of solipeds causing severe disease in animals and men. Although eradicated from many Western countries, it recently emerged in Asia, the Middle-East, Africa, and South America. Due to its rareness, little is known about outbreak dynamics of the disease and its epidemiology. We investigated a recent outbreak of glanders in Bahrain by applying high resolution genotyping (multiple locus variable number of tandem repeats, MLVA) and comparative whole genome sequencing to B. mallei isolated from infected horses and a camel. These results were compared to samples obtained from an outbreak in the United Arab Emirates in 2004, and further placed into a broader phylogeographic context based on previously published B. mallei data. The samples from the outbreak in Bahrain separated into two distinct clusters, suggesting a complex epidemiological background and evidence for the involvement of multiple B. mallei strains. Additionally, the samples from Bahrain were more closely related to B. mallei isolated from horses in the United Arab Emirates in 2004 than other B. mallei which is suggestive of repeated importation to the region from similar geographic sources. High-resolution genotyping and comparative whole genome analysis revealed the same phylogenetic patterns among our samples. The close relationship of the Dubai/UAE B. mallei populations to each other may be indicative of a similar geographic origin that has yet to be identified for the infecting strains. The recent emergence of glanders in combination with worldwide horse trading might pose a new risk for human infections.


Per2 participates in AKT-mediated drug resistance in A549/DDP lung adenocarcinoma cells.
Period2 (Per2) is a key mammalian circadian clock protein, and additionally has a tumor suppressive function. The present study aimed to investigate its role in drug resistance in A549/cisplatin (DDP) lung adenocarcinoma cells. Per2 knockdown and overexpression in A549/DDP cells were used to compare cell proliferation (by MTT assay), apoptosis (active-caspase 3 western blot) and clone forming assay. The activation of AKT/mechanistic target of rapamycin (mTOR) was investigated by a western blot assay. The Per2 expression level was decreased in A549/DDP cells compared with A549 cells. Per2 knockdown by short hairpin RNA protects A549/DDP cells from apoptosis, and promotes proliferation and migration. Per2 knockdown results in increased activation of the phosphoinositide 3-kinase (PI3K)/AKT/mTOR signaling pathway. Overexpression of Per2 in A549/DDP cells may reduce the activity of the PI3K/AKT/mTOR signaling pathway, and promote apoptosis of A549 cells. The results of the present study suggest that Per2 participates in AKT-mediated drug resistance in A549/DDP lung adenocarcinoma cells.


Resistance to activated protein C as a risk factor of stroke in a thalassemic patient.
It is well known that thalassemic patients exhibit an increased frequency of thrombotic events. Most individuals with resistance to activated protein C (APCR) are the result of a point mutation replacing Arg 506 with Gln in the factor V aminoacidic sequence (factor V Leiden). Recently APCR has been shown to account for up to 50% of cases of thrombophilia. In this report, we describe a 10 year old thalassemic intermedia patient heterozygous for Factor V R506Q who developed a stroke following transfusion. Coagulation laboratory values were all within the normal range and there was no evidence of a lupus anticoagulant. Computerized brain tomography showed an ischemic area in the left temporo-parietal region. At follow-up, plasma from the patient demonstrated APCR and molecular diagnostic testing revealed heterozygosity for factor V R506Q. We suggest that the heterozygosity for factor V Leiden could increase the thrombotic risk in thalassemia intermedia. We believe it may be beneficial to screen all intermedia thalassemic patients for APCR especially before starting a transfusional regimen.


Different neural manifestations of two slow frequency bands in resting functional magnetic resonance imaging: a systemic survey at regional, interregional, and network levels.
Temporal and spectral perspectives are two fundamental facets in deciphering fluctuating signals. In resting state, the dynamics of blood oxygen level-dependent (BOLD) signals recorded by functional magnetic resonance imaging (fMRI) have been proven to be strikingly informative (0.01-0.1 Hz). The distinction between slow-4 (0.027-0.073 Hz) and slow-5 (0.01-0.027 Hz) has been described, but the pertinent data have never been systematically investigated. This study used fMRI to measure spontaneous brain activity and to explore the different spectral characteristics of slow-4 and slow-5 at regional, interregional, and network levels, respectively assessed by regional homogeneity (ReHo) and mean amplitude of low-frequency fluctuation (mALFF), functional connectivity (FC) patterns, and graph theory. Results of paired t-tests supported/replicated recent research dividing low-frequency BOLD fluctuation into slow-4 and slow-5 for ReHo and mALFF. Interregional analyses showed that for brain regions reaching statistical significance, FC strengths at slow-4 were always weaker than those at slow-5. Community detection algorithm was applied to FC data and unveiled two modules sensitive to frequency effects: one comprised sensorimotor structure, and the other encompassed limbic/paralimbic system. Graph theoretical analysis verified that slow-4 and slow-5 differed in local segregation measures. Although the manifestation of frequency differences seemed complicated, the associated brain regions can be grossly categorized into limbic/paralimbic, midline, and sensorimotor systems. Our results suggest that future resting fMRI research addressing the three above systems either from neuropsychiatric or psychological perspectives may consider using spectrum-specific analytical strategies.


Structural basis for α-bungarotoxin insensitivity of neuronal nicotinic acetylcholine receptors.
The ten types of nicotinic acetylcholine receptor α-subunits show substantial sequence homology, yet some types confer high affinity for α-bungarotoxin, whereas others confer negligible affinity. Combining sequence alignments with structural data reveals three residues unique to α-toxin-refractory α-subunits that coalesce within the 3D structure of the α4β2 receptor and are predicted to fit between loops I and II of α-bungarotoxin. Mutating any one of these residues, Lys189, Ile196 or Lys153, to the α-toxin-permissive counterpart fails to confer α-bungarotoxin binding. However, mutating both Lys189 and Ile196 affords α-bungarotoxin binding with an apparent dissociation constant of 104 nM, while combining mutation of Lys153 reduces the dissociation constant to 22 nM. Analogous residue substitutions also confer high affinity α-bungarotoxin binding upon α-toxin-refractory α2 and α3 subunits. α4β2 receptors engineered to bind α-bungarotoxin exhibit slow rates of α-toxin association and dissociation, and competition by cholinergic ligands typical of muscle nicotinic receptors. Receptors engineered to bind α-bungarotoxin co-sediment with muscle nicotinic receptors on sucrose gradients, and mirror single channel signatures of their α-toxin-refractory counterparts. Thus the inability of α-bungarotoxin to bind to neuronal nicotinic receptors arises from three unique and interdependent residues that coalesce within the receptor's 3D structure.


[Wilson-Konovalov disease in 3 sisters: a radical change in prognosis if timely diagnosed].
Wilson-Konovalov disease is a rare autosomal recessive genetic disorder in which copper accumulates in the liver, brain and other target organs. The paper describes a family case of the abdominal form of the disease in three sisters, the eldest of them died from fulminant liver failure at the age of 18 years. The second sister aged 16 years was diagnosed as having the disease at the stage of decompensated liver cirrhosis; her treatment with D-penicillamine resulted in complete disease remission. The youngest sister was diagnosed with the disease at the preclinical stage, which could expect its good prognosis. However, the patient's refusal of treatment led to death from liver failure. This case demonstrates the importance of timely diagnosis and the possibility of dramatic improvement in prognosis even at the stage of decompensated liver cirrhosis.


Closing the gap in paediatric ventricular assist device therapy with the Berlin Heart EXCOR® 15-ml pump.
The Berlin Heart EXCOR ® (EXCOR) paediatric ventricular assist device is used worldwide for mechanical support of infants and small children with end-stage heart failure. A clinically important gap between the smallest EXCOR blood pump (10 ml) and the next larger size (25 ml) limited the choice of pump size in patients with a body surface area (BSA) between 0.33 and 0.5 m 2 . We present the first clinical experience from the early product surveillance (EPS) of the new EXCOR 15-ml blood pump. After CE and U.S. Food and Drug Administration approval in January 2013, 20 patients with a mean age of 1.6 years (range 0.5-3.5 years) and a mean BSA of 0.45 m 2 (range 0.33-0.59 m 2 ) were enrolled in the EPS. The main diagnosis was idiopathic cardiomyopathy in 13 patients; the majority ( n = 16) of children were in INTERMACS level 1 or 2. Data from high-volume paediatric transplant centres were collected prospectively for a defined follow-up period of 60 days after device implantation. Mean time on the EXCOR 15-ml blood pump was 43 days; the survival rate was 100% at the end of the EPS period. Seven patients underwent a heart transplant from the device; 2 children were weaned; and 11 patients remained on support. Infection of cannula exit sites occurred in 3 patients. Two patients had minor thromboembolic strokes but made a complete neurological recovery. The new EXCOR 15-ml blood pump demonstrated optimal ventricular assist device support of children with a BSA of 0.33-0.5 m 2 .


Assessment of fluoride levels during pregnancy and its association with early adverse pregnancy outcomes.
There is sparse data available on human subjects regarding the affect of excessive fluoride exposures on pregnancy. The aim of this study was to examine the association between elevated urinary fluoride levels during early pregnancy and maternal anemia and adverse fetal outcome. We enrolled 600 pregnant patients with gestational age less than 20 weeks and with a high urinary fluoride levels(>1 mg/L).We also documented the fluoride levels in the tap water and ground water samples collected from the areas where these women resided during pregnancy. These patients were also evaluated for hemoglobin levels and detailed fetal examination by ultrasound. Data was assessed by SSPS version 16.0 software and P < 0.05 was considered to be significant. Urinary fluoride averaged 2.65 mg/L and ranged from 1.0 to 4.3 mg/L while all the water samples revealed fluoride levels greater than WHO prescribed the limit of 1.5 mg/L. A total of 402 patients (67%) were found to be anemic with hemoglobin levels ranging from 6.2 to 11.9 g/dl (9.28 ± 1.29). Eighty one patients (13.5%) had adverse fetal outcomes that comprised abortions, congenital abnormalities, and intrauterine deaths (IUDs). There was a negative correlation between urinary fluoride and hemoglobin levels (P = 0.031, r= -0.59) and females with elevated urinary fluoride levels were found to have a strong association with the pregnancy complications, i.e., anemia, miscarriage, abortion, and still birth (χ2 = 9.23, P < 0.05). Excess fluoride exposures can have deleterious effects on the expecting mother and fetus and is associated with adverse pregnancy outcomes.


Pain and its clinical associations in individuals with COPD: a systematic review.
Pain is emerging as a clinical complication in COPD, but the clinical impact of this comorbidity and the measurement properties of instruments used to assess pain require evaluation. Electronic searches of five databases were performed up to September 2014 for the two phases of this review. To be included in phase 1, studies reported the clinical associations of pain and prevalence in individuals with COPD. To be included in phase 2, studies reported measurement properties of an instrument assessing pain in COPD. Two independent reviewers rated the quality of quantitative and qualitative evidence (phase 1) and the measurement properties using the four-point Consensus-Based Standards for the Selection of Health Status Measurement Instruments (COSMIN) checklist (phase 2). Of the 358 studies identified in the literature, nine met the inclusion criteria for phase 1 and five for phase 2. The mean (SD) quality score (of 16) for the quantitative studies was 13.1 (1.7). The pooled prevalence of pain in moderate to very severe COPD was 66% (95% CI, 44%-85%). Higher pain intensity was associated with increased dyspnea, fatigue, poorer quality of life, and a greater quantity of specific comorbidities. Of the two identified instruments (Brief Pain Inventory and McGill Pain Questionnaire), the measurement properties analyzed were construct validity, internal consistency, and criterion-predictive validity, with variable findings based on "fair" or "poor" quality studies. In people with COPD, pain has negative clinical associations with symptoms and quality-of-life measures. Further research exploring the measurement properties of instruments assessing pain is required.


Transmission through chiral twist defects in anisotropic periodic structures.
A long-lived photonic state is observed in measurements of microwave transmission through a helical stack of anisotropic overhead transparencies with various twist defects in the center of the structure. Once account is taken of absorption and of the angular spread of the source, computer simulations of transmission through a polarized localized state are in agreement with measurements. Unlike for isotropic one-dimensional bandgaps, the intensity of the localized mode is not modulated in space on a wavelength scale.


Genetic changes caused by restocking and hydroelectric dams in demographically bottlenecked brown trout in a transnational subarctic riverine system.
Habitat discontinuity, anthropogenic disturbance, and overharvesting have led to population fragmentation and decline worldwide. Preservation of remaining natural genetic diversity is crucial to avoid continued genetic erosion. Brown trout (Salmo trutta L.) is an ideal model species for studying anthropogenic influences on genetic integrity, as it has experienced significant genetic alterations throughout its natural distribution range due to habitat fragmentation, overexploitation, translocations, and stocking. The Pasvik River is a subarctic riverine system shared between Norway, Russia, and Finland, subdivided by seven hydroelectric power dams that destroyed about 70% of natural spawning and nursing areas. Stocking is applied in certain river parts to support the natural brown trout population. Adjacent river segments with different management strategies (stocked vs. not stocked) facilitated the simultaneous assessment of genetic impacts of dams and stocking based on analyses of 16 short tandem repeat loci. Dams were expected to increase genetic differentiation between and reduce genetic diversity within river sections. Contrastingly, stocking was predicted to promote genetic homogenization and diversity, but also potentially lead to loss of private alleles and to genetic erosion. Our results showed comparatively low heterozygosity and clear genetic differentiation between adjacent sections in nonstocked river parts, indicating that dams prevent migration and contribute to genetic isolation and loss of genetic diversity. Furthermore, genetic differentiation was low and heterozygosity relatively high across stocked sections. However, in stocked river sections, we found signatures of recent bottlenecks and reductions in private alleles, indicating that only a subset of individuals contributes to reproduction, potentially leading to divergence away from the natural genetic state. Taken together, these results indicate that stocking counteracts the negative fragmentation effects of dams, but also that stocking practices should be planned carefully in order to ensure long-term preservation of natural genetic diversity and integrity in brown trout and other species in regulated river systems.


Implementation of the ANISA Protocol in Sylhet, Bangladesh: Challenges and Solutions.
Despite the high rate of deaths in young infants (0-59 days) attributable to infections in resource-poor countries, data on bacterial and viral etiologies of community-acquired infections in this age group are limited. These data are needed to develop appropriate preventive strategies and suitable antibiotic treatment regimens for reducing the number of young infant deaths from infections. The Aetiology of Neonatal Infection in South Asia (ANISA) study is designed to generate these critical data and is being implemented in Bangladesh, India and Pakistan. The Sylhet site in Bangladesh was selected because neonatal mortality is high in this country and particularly in Sylhet District. In this article, we describe the contextual challenges in implementing the ANISA study in Sylhet, as well as the strategies developed by our team to address these challenges. The major challenge in implementing the ANISA protocol in Sylhet is conducting the first postnatal visit within 24 hours of birth. This problem stems from several social, cultural and geographical characteristics of the study population and its demographic profile. In this area, most births take place at home, referral compliance for newborn illness to health facilities is low and the blood culture contamination rate is high. Community mobilization, cellphone-based birth notification by families, delivery of quality services at study hospitals and referral support to families in need were some of the strategies adopted by the Sylhet site team for overcoming these challenges during study implementation. Quality control in specimen collection, transportation and processing also plays a role in ensuring satisfactory performance. Our research team, with support from the ANISA coordination center, has successfully addressed these challenges and is implementing the study protocol while maintaining the high quality benchmark set by the coordination center.


[DNA damages in individual mammalian cells].
The method of alkaline microelectrophoresis of nucleoid DNA of isolated cells built in an agar gel was used to study the harmful effect of radiation on the genome of mouse peripheral blood lymphocytes. The yield of the lymphocyte DNA structure damages in irradiated animals and in the in vitro exposed lymphocytes was a linear function of radiation dose within the dose range from 0.5 to 5 Gy and 0.27 to 1 Gy respectively. A study was made of the dynamics of DNA structure damages in cells NIH 3T3 exposed to 3 Gy radiation, and in mouse lymphocytes exposed to 4 Gy radiation.


[Quantitative characteristics of the vestibular nuclei of the brain in the dolphin and man].
Selective qualitative and quantitative distinctions, most markedly expressed in dolphins as compared with man, were found in the lateral vestibular nucleus of the brain. In man it is the least nucleus (according to the territory) in the whole vestibular complex, while in dolphins it is the largest nucleus (9-16 times as great as in man), which has the largest area but the least density of cells. The above selectivity of changes in the structural organization of the lateral vestibular nucleus is considered as a result of ecological and physiological differences in the organization and coordination of the volume of motor acts and muscle tone when changing the position of the body in space.


Predictive value of tumor recurrence using urinary vascular endothelial factor levels in patients receiving radiation therapy for Glioblastoma Multiforme (GBM).
Glioblastoma Multiforme (GBM) is the most common primary malignant tumor of the central nervous system. Standard of care includes maximal resection followed by chemoradiotherapy. Tumors need adequate perfusion and neovascularization to maintain oxygenation and for removal of wastes. Vascular endothelial growth factor (VEGF) is a well characterized pro-angiogenic factor. We hypothesized that the increases in urinary VEGF levels would occur early in the course of tumor recurrence or progression. We examine the feasibility of collecting and analyzing urinary VEGF levels in a prospective, multi-institutional trial (Radiation Therapy Oncology Group, RTOG, 0611) as well as the role of VEGF as a marker of tumor recurrence. We evaluated VEGF levels in urine specimens collected post-operatively, at the conclusion of radiation therapy (RT) and one month following RT. Urinary VEGF levels were correlated with tumor progression at one year. VEGF levels were measured by enzyme-linked immunosorbant assay in urine specimens and normalized to urinary creatinine levels. Sample size was determined based on a 50% 1-year recurrence rate. With a sensitivity and specificity of 80%, the expected 95% confidence interval was (0.69, 0.91) with 100 patients. A failure was defined as documented disease progression, recurrence or death before one year. 202 patients were enrolled between February-2006 and October-2007. Four patients were ineligible as they did not receive RT. Of the remaining 198 patients, 128 had all three samples collected. In this group, 35 patients (27.3%) did not progress, 89 (69.5%) had progression and 4 (3.1%) died without evidence of progression. Median VEGF levels at baseline were 52.9 pg/mg Cr (range 0.2- 15,034.4); on the last day of RT, 56.6 (range 0-2,377.1); and at one month follow-up, 70.0 (range 0.1-1813.2). In patients without progression at 1-year, both baseline VEGF level and end of RT VEGF level were lower than those of patients who progressed: 40.3 (range 0.2-350.8) vs. 59.7 (range 1.3-15,034.4) and 41.8 (range 0-356.8) vs. 69.7 (range 0-2,377.1), respectively. This did not reach statistical significance. Comparison of the change in VEGF levels between the end of RT and one month following RT, demonstrated no significant difference in the proportions of progressors or non-progressors at 1-year for either the VEGF increased or VEGF decreased groups. Urine can be collected and analyzed in a prospective, multi-institutional trial. In this study of patients with GBM a change in urinary VEGF levels between the last day of RT and the one month following RT did not predict for tumor progression by one year.


[Expression of UCP3 and the sensitivity of mitochondrial permeability transition pore opening to Ca2+ in old rat heart under activation of biosynthesis of coenzyme Q].
The expression of mitochondrial uncoupling protein 3 (UCP3), as well as the sensitivity of mitochondrial permeability transition pore opening (MPTP) to Ca2+ (10(-4) mol/l) in old rat heart under activation in vivo of ubiquinone synthesis--coenzyme Q, (CoQ) via administration of the precursors (4-hydroxybenzoic acid, aminoacid methionine and modulator vitamin E) were studied. It was shown that the expression level of UCP3 decreased by 63% in old rats compared to adult rats and this was accompanied by an increased sensitivity of the MPT to calcium. Under activation of endogenous synthesis of CoQ it was observed almost complete restoration of UCP3 expression in old rat heart and a decrease in the sensitivity of the MPTP opening to Ca2+. In mitochondria from old rat hearts we noted an increased content of the superoxide (O2) and hydroxyl (OH) radicals and of the stable metabolite of active oxygen species hydrogen peroxide (H2O2), as compared to those in adult animals. Following activation of endogenous synthesis of CoQ in old rat heart mitochondria it was observed a decreased content of H2O2, and the tendency for decreasing the levels of the radicals O2 and MOH. The results obtained allowed to conclude that the CoQ-dependent restoration of the UCP3 levels in old rat heart and antioxidant/cardioprotective effects of CoQ related to the MPTP opening inhibition can reduce the oxidative stress and thus prevent the manifestation of mitochondrial dysfunction in aging heart. We suggest that UCP3 is not involved in the increase of the passive H-conductance through the inner mitochondrial membrane in the aging heart, and that CoQ as a factor of respiratory chain could be an important endogenous regulator of the uncoupling proteins, in particular UCP3, in the heart.


Anti-oxidative and anti-genotoxic effects of carnosine on human lymphocyte culture.
The aim of this study was to investigate the effects of carnosine, a biological antioxidant, on the oxidative stress and genotoxicity by a single dose of carbon tetrachloride (CCl(4); 5 mM) in the human lymphocyte culture. We studied the anti-genotoxic effects of carnosine by using sister chromatid exchange (SCE) test system. Also, the anti-oxidative effects of carnosine were evaluated by using superoxide dismutase (SOD), glutathione peroxidase (GPx), total glutathione (GSH) and malondialdehyde (MDA) assay. The SCE frequency was increased when treated with CCl(4). Carnosine at 10 and 20 mM reduced SCE frequency in the human lymphocyte (p < 0.001). In addition, CCl(4) treatment significantly depleted the level of GSH, reduced the activity of SOD and GPx and elevated the level of MDA (p < 0.001). Carnosine treatment led to significant attenuation of CCl(4)-induced oxidative stress by normalization of the activities of SOD and GPx and the level of GSH and MDA (p < 0.05 or 0.001). These results suggest that carnosine could provide anti-oxidative and anti-genotoxic protection for the oxidative and genotoxic agents that cause many diseases including cancer and neurodegenerative disease.


A preliminary investigation of tuberculosis and other diseases in African buffalo (Syncerus caffer) in Queen Elizabeth National Park, Uganda.
A survey to determine the prevalence of bovine tuberculosis caused by Mycobacterium bovis and certain other infectious diseases was conducted on 42 free-ranging African buffaloes, (Syncerus caffer) from May to June 1997 in the Queen Elizabeth National Park, Uganda. Using the gamma interferon test, exposure to M. bovis was detected in 21.6% of the buffaloes. One dead buffalo and an emaciated warthog (Phacochoerus aethiopicus) that was euthanased, were necropsied; both had miliary granulomas from which M. bovis was isolated. None of the buffaloes sampled in Sector A of the park, which has no cattle interface, tested positive for bovine tuberculosis (BTB) exposure. The prevalence and distribution of BTB does not appear to have changed significantly since the 1960s, but this may be due to fluxes in the buffalo population. Serological testing for foot-and-mouth disease (FMD) demonstrated positive exposure of 57.1% of the buffaloes sampled, with types A, O and SAT 1-3, which is the first known report of FMD antibodies to A and O types in free ranging African buffaloes. Foot-and-mouth disease virus types SAT 1 and SAT 3 were isolated from buffalo probang samples. Two percent of the buffaloes had been exposed to brucellosis. None of the buffaloes tested had antibodies to rinderpest, leptospirosis or Q fever.


FOXO3a loss is a frequent early event in high-grade pelvic serous carcinogenesis.
Serous ovarian carcinoma is the most lethal gynecological malignancy in Western countries. The molecular events that underlie the development of the disease have been elusive for many years. The recent identification of the fallopian tube secretory epithelial cells (FTSECs) as the cell-of-origin for most cases of this disease has led to studies aimed at elucidating new candidate therapeutic pathways through profiling of normal FTSECs and serous carcinomas. Here we describe the results of transcriptional profiles that identify the loss of the tumor suppressive transcription factor FOXO3a in a vast majority of high-grade serous ovarian carcinomas. We show that FOXO3a loss is a hallmark of the earliest stages of serous carcinogenesis and occurs both at the DNA, RNA and protein levels. We describe several mechanisms responsible for FOXO3a inactivity, including chromosomal deletion (chromosome 6q21), upregulation of miRNA-182 and destabilization by activated PI3K and MEK. The identification of pathways involved in the pathogenesis of ovarian cancer can advance the management of this disease from being dependant on surgery and cytotoxic chemotherapy alone to the era of targeted therapy. Our data strongly suggest FOXO3a as a possible target for clinical intervention.


Tetraarsenic oxide-induced inhibition of malignant glioma cell invasion in vitro via a decrease in matrix metalloproteinase secretion and protein kinase B phosphorylation.
Local invasiveness of malignant glioma is a major reason for the failure of current treatments including surgery and radiation therapy. Tetraarsenic oxide (As4O6 [TAO]) is a trivalent arsenic compound that has potential anticancer and antiangiogenic effects in selected cancer cell lines at a lower concentration than arsenic trioxide (As2O3 [ATO]), which has been more widely tested in vitro and in vivo. The authors tried to determine the cytotoxic concentration of TAO in malignant glioma cell lines and whether TAO would show anti-invasive effects under conditions independent of cell death or apoptosis. The human phosphatase and tensin homolog (PTEN)-deficient malignant glioma cell lines U87MG, U251MG, and U373MG together with PTEN-functional LN428 were cultured with a range of micromolar concentrations of TAO. The invasiveness of the glioma cell lines was analyzed. The effect of TAO on matrix metalloproteinase (MMP) secretion and membrane type 1 (MT1)-MMP expression was measured using gelatin zymography and Western blot, respectively. Akt, or protein kinase B, activity, which is a downstream effector of PTEN, was assessed with a kinase assay using glycogen synthesis kinase-3β (GSK-3β) as a substrate and Western blotting of phosphorylated Akt. Tetraarsenic oxide inhibited 50% of glioma cell proliferation at 6.3-12.2 μM. Subsequent experiments were performed under the same TAO concentrations and exposure times, avoiding the direct tumoricidal effect of TAO, which was confirmed with apoptosis markers. An invasion assay revealed a dose-dependent decrease in invasiveness under the influence of TAO. Both the gelatinolytic activity of MMP-2 and MT1-MMP expression decreased in a dose-dependent manner in all cell lines, which was in accordance with the invasion assay results. The TAO decreased kinase activity of Akt on GSK-3β assay and inhibited Akt phosphorylation in a dose-dependent manner in all cell lines regardless of their PTEN status. These results showed that TAO effectively inhibits proliferation of glioblastoma cell lines and also exerts an anti-invasive effect via decreased MMP-2 secretion, decreased MT1-MMP expression, and the inhibition of Akt phosphorylation under conditions devoid of cytotoxicity. Further investigations using an in vivo model are needed to evaluate the potential role of TAO as an anti-invasive agent.


Intermittent cyclophosphamide in refractory rheumatoid arthritis.
Three patients with refractory rheumatoid arthritis were treated with oral cyclophosphamide; in two cases this was supplemented with pulse treatment with methylprednisolone. Long term remission was induced in all three patients and was sustained until follow up at least nine months after the methylprednisolone was stopped. Leucopenia occurred but resolved when cyclophosphamide was reduced from daily to intermittent dosing. Intermittent treatment with cyclophosphamide, possibly in conjunction with pulses of methylprednisolone, may induce remission in patients with rheumatoid arthritis refractory to other forms of treatment.