Increased expression of metallothionein is associated with irinotecan resistance in gastric cancer.
To gain insight into clinically relevant mechanisms of irinotecan resistance, we undertook oligonucleotide microarray analyses on paired malignant effusion samples obtained from eight gastric cancer patients treated with weekly irinotecan. Pretreatment and posttreatment (48 h) effusion samples were obtained for each patient, and the change in expression profile was compared between clinical responders and nonresponders. When differences in the expression of genes were examined using SAM (Significance Analysis of Microarrays) software, five isoforms of the metallothionein family were identified to have significantly higher signal log ratios in five nonresponders, compared with three responders. Compared with control cells, metallothionein 1X (MT1X)-transfected AGS cells showed a 1.4-fold higher irinotecan IC(50) by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and tended to form more colonies. These findings collectively suggest that irinotecan-induced up-regulation of metallothionein might be associated with irinotecan resistance in patients with gastric cancer, although it remains to be confirmed in a larger data set.


The upper respiratory tract microbiota of healthy adults is affected by Streptococcus pneumoniae carriage, smoking habits, and contact with children.
The microbiota of the upper respiratory tract is increasingly recognized as a gatekeeper of respiratory health. Despite this, the microbiota of healthy adults remains understudied. To address this gap, we investigated the composition of the nasopharyngeal and oropharyngeal microbiota of healthy adults, focusing on the effect of Streptococcus pneumoniae carriage, smoking habits, and contact with children. Differential abundance analysis indicated that the microbiota of the oropharynx was significantly different from that of the nasopharynx (P < 0.001) and highly discriminated by a balance between the classes Negativicutes and Bacilli (AUC of 0.979). Moreover, the oropharynx was associated with a more homogeneous microbiota across individuals, with just two vs. five clusters identified in the nasopharynx. We observed a shift in the nasopharyngeal microbiota of carriers vs. noncarriers with an increased relative abundance of Streptococcus, which summed up to 30% vs. 10% in noncarriers and was not mirrored in the oropharynx. The oropharyngeal microbiota of smokers had a lower diversity than the microbiota of nonsmokers, while no differences were observed in the nasopharyngeal microbiota. In particular, the microbiota of smokers, compared with nonsmokers, was enriched (on average 16-fold) in potential pathogenic taxa involved in periodontal diseases of the genera Bacillus and Burkholderia previously identified in metagenomic studies of cigarettes. The microbiota of adults with contact with children resembled the microbiota of children. Specifically, the nasopharyngeal microbiota of these adults had, on average, an eightfold increase in relative abundance in Streptococcus sp., Moraxella catarrhalis, and Haemophilus influenzae, pathobionts known to colonize the children's upper respiratory tract, and a fourfold decrease in Staphylococcus aureus and Staphylococcus lugdunensis. Our study showed that, in adults, the presence of S. pneumoniae in the nasopharynx is associated with a shift in the microbiota and dominance of the Streptococcus genus. Furthermore, we observed that smoking habits are associated with an increase in bacterial genera commonly linked to periodontal diseases. Interestingly, our research also revealed that adults who have regular contact with children have a microbiota enriched in pathobionts frequently carried by children. These findings collectively contribute to a deeper understanding of how various factors influence the upper respiratory tract microbiota in adults. Video Abstract.


Clinical significance of serum miR-21 in breast cancer compared with CA153 and CEA.
MicroRNA-21 (miR-21) has been shown to be a key regulator of carcinogenesis. There were few reports about the comparison of serum miR-21 with conventional tumor markers. This study aimed to explore the diagnostic value of circulating miR-21 as a tumor marker in breast cancer (BC) and compare it with CA153 and carcinoembryonic antigen (CEA). Circulating miR-16 and miR-21 were amplified and quantitatively detected by real-time PCR in 89 BC patients and 55 healthy controls. The levels of CA153 and CEA were measured through electrochemiluminescence assays. Then the sensitivity in diagnosis of BC was compared among miR-21, CA153 and CEA. The level of serum miR-21 was significantly higher in BC patients than controls (P<0.001). The sensitivity and specificity of miR-21 were 87.6% and 87.3%, respectively, whereas the sensitivities of CEA and CA153 were only 22.47% and 15.73%. Compared with CEA and CA153, serum miR-21 has a higher sensitivity in diagnosis of BC. Although not correlated with the status of ER, PR and clinical stages, serum miR-21 may be a potential diagnostic indicator for BC, especially for the early stage.


The fate of threatened coastal dune habitats in Italy under climate change scenarios.
Coastal dunes worldwide harbor threatened habitats characterized by high diversity in terms of plant communities. In Italy, recent assessments have highlighted the insufficient state of conservation of these habitats as defined by the EU Habitats Directive. The effects of predicted climate change could have dramatic consequences for coastal environments in the near future. An assessment of the efficacy of protection measures under climate change is thus a priority. Here, we have developed environmental envelope models for the most widespread dune habitats in Italy, following two complementary approaches: an "indirect" plant-species-based one and a simple "direct" one. We analyzed how habitats distribution will be altered under the effects of two climate change scenarios and evaluated if the current Italian network of protected areas will be effective in the future after distribution shifts. While modeling dune habitats with the "direct" approach was unsatisfactory, "indirect" models had a good predictive performance, highlighting the importance of using species' responses to climate change for modeling these habitats. The results showed that habitats closer to the sea may even increase their geographical distribution in the near future. The transition dune habitat is projected to remain stable, although mobile and fixed dune habitats are projected to lose most of their actual geographical distribution, the latter being more sensitive to climate change effects. Gap analysis highlighted that the habitats' distribution is currently adequately covered by protected areas, achieving the conservation target. However, according to predictions, protection level for mobile and fixed dune habitats is predicted to drop drastically under the climate change scenarios which we examined. Our results provide useful insights for setting management priorities and better addressing conservation efforts to preserve these threatened habitats in future.


A rare case of clear-cell carcinoid of appendix.
The carcinoids are the most frequent type of tumors arising from the appendix. In the majority of cases, these tumors are asymptomatic and usually are discovered after appendectomy. Definitive diagnosis relies on pathological examination of the resected appendix, size of the tumor being critical for the further management. Clear-cell change in neuroendocrine tumors (NETs) has rarely been described in the appendix. We choose to present a clear-cell carcinoid subtype of appendiceal NET to raise awareness on this potentially curable and rare condition that can be overlooked. We highlight the importance of the pathological exam and the morphological and immunohistochemical behavior of the tumor in confirming the diagnosis and aiding in the treatment decision making. Also, important entities should be considered in the process of differential diagnosis such as goblet-cell carcinoid or renal-cell÷ovarian carcinoma.


Herd immunity: recent uses in vaccine assessment.
Human communities defend themselves against specific infectious agents in a way that extends beyond the simple sum of the immune status of its individuals. By analogy with individual immunity to specific agents, the community level of immunity may vary from complete susceptibility to full protection. Herd immunity has been used to name this community property, which is the result of evolution through natural selection, leading to relationships between two species, typical of prey-predator systems. Varying uses of the term herd immunity led to the use of other expressions, such as herd protection, herd effect and community immunity. Knowledge derived from observational studies and models on herd immunity has supported decisions on the choice of vaccines and vaccination strategies for the benefit of populations. This knowledge is most likely to be extended in the future, with far-reaching effects.


Human cytomegalovirus UL38 protein blocks apoptosis.
Apoptosis is an innate cellular defense response to viral infection. The slow-replicating human cytomegalovirus (HCMV) blocks premature death of host cells prior to completion of the infection cycle. In this study, we report that the HCMV UL38 gene encodes a cell death inhibitory protein. A mutant virus lacking the pUL38 coding sequence, ADdlUL38, grew poorly in human fibroblasts, failed to accumulate viral DNA to wild-type levels, and induced excessive death of infected cells. Cells expressing pUL38 were resistant to cell death upon infection and effectively supported the growth of ADdlUL38. Cells infected with the pUL38-deficient virus showed morphological changes characteristic of apoptosis, including cell shrinkage, membrane blebbing, vesicle release, and chromatin condensation and fragmentation. The proteolytic cleavage of two key enzymes involved in apoptosis, namely, caspase 3 and poly(ADP-ribose) polymerase, was activated upon ADdlUL38 infection, and the cleavage was blocked in cells expressing pUL38. The pan-caspase inhibitor Z-VAD-FMK largely restored the growth of ADdlUL38 in normal fibroblasts, indicating that the defective growth of the mutant virus mainly resulted from premature death of host cells. Furthermore, cells expressing pUL38 were resistant to cell death induced by a mutant adenovirus lacking the antiapoptotic E1B-19K protein or by thapsigargin, which disrupts calcium homeostasis in the endoplasmic reticulum. Taken together, these results indicate that the HCMV protein pUL38 suppresses apoptosis, blocking premature death of host cells to facilitate efficient virus replication.


Effect of alcohol coadministration on the plasma and brain concentrations of cocaine in rats.
We investigated the effect of intravenous alcohol coadministration on the pharmacokinetics of cocaine in awake, freely moving rats using the microdialysis technique. Alcohol coadministration resulted in faster rate of cocaine absorption after intraperitoneal administration leading to higher cocaine plasma concentration. The higher plasma cocaine concentration resulted in a proportional increase in the cocaine brain extracellular fluid concentration. However, cocaine brain extracellular fluid/plasma distribution ratio, determined from the ratio of the corresponding cocaine area under the concentration-time curves, was not affected by alcohol coadministration. Cocaethylene was detected only after administration of cocaine + alcohol. The brain extracellular fluid/plasma distribution ratio of cocaethylene was similar to that of cocaine. The higher cocaine concentrations in plasma and brain extracellular fluid, in addition to the formation of the pharmacologically active metabolite cocaethylene are, at least partially, responsible for the increased cocaine effects produced after administration of this drug combination.


Premutilin Synthase: Ring Rearrangement by a Class II Diterpene Cyclase.
Biosynthesis of the complex diterpenoid antibiotic pleuromutilin relies on a bifunctional (di)terpene synthase, and here site-directed mutagenesis was used to knockout either of the two active sites. This enabled characterization of the novel ring contracted intermediate produced by the initiating class II diterpene cyclase active site. Quantum chemical calculations further indicate the importance of reactant configuration for this intriguing ring rearrangement.


Dihydroartemisinin (DHA) treatment causes an arrest of cell division and apoptosis in rat embryonic erythroblasts in whole embryo culture.
Within 24 hr after oral administration of the antimalarial artesunate to rats on Day 10 or 11 postcoitum (pc), there is depletion of embryonic erythroblasts (EEbs), leading to embryo malformation and death. The proximate agent is dihydroartemisinin (DHA), the primary metabolite. We investigated the causes of EEb depletion by evaluating effects of DHA on EEbs in whole embryo culture (WEC). Rat embryos cultured starting on Day 9 pc were treated with 1 or 7 μM DHA for 24 hr starting after 19 hr of culture (∼Day 10 pc) and for 2 to 12 hr starting after 43 hr of culture (∼Day 11 pc). DHA effects indicating the depletion of EEbs were paling of the visceral yolk sac and reductions in visible blood cells, H&E-stained normal (Type II or III) EEbs, and dividing (BrdU-stained) EEbs. DHA-induced abnormal cell division was indicated by increases in symmetric and asymmetric binuclear cells. DHA-induced apoptosis was indicated by increases in TUNEL- and Caspase-3-positive cells and EEbs with fragmented nuclei. In addition, although the overall number of EEbs was decreasing, DHA caused increases in the numbers of circulating early-stage (Type I or earlier) EEbs that could not be accounted for by cell division, suggesting the release of new, less sensitive erythroblasts from the yolk sac. In summary, treatment of Day 10 or 11 pc rat embryos with DHA in WEC resulted in defective and arrested cell division in EEbs followed by apoptosis, suggesting a mechanism for their depletion after artesunate treatment in vivo.


The future in advanced prostate cancer: take your partners or is the last dance for me?
Recent therapeutic initiatives have improved quality of life and survival for patients with advanced prostate cancer. This review focuses predominantly on prostate cancer that has become refractory to standard androgen ablation treatment. Planned trials will answer further questions on the optimal use and sequencing of currently available hormonal agents, cytotoxic therapies, and radiolabeled nucleotides. Future therapeutic advances are likely to come in 2 areas: targeted therapies and response prediction. Molecular targeted agents will be most useful in combination with each other or with established systemic therapies. The selection of combinations will require the application of paradigms targeting key biochemical pathways and specific microenvironments in prostate cancer. Response prediction for individual patients may be assisted by either pretreatment or sequential molecular profiling, or sequential imaging, or biochemical studies that predicate outcome prior to or soon after treatment has been initiated. To bring these advances to the metastatic prostate cancer patient, a series of well-designed clinical trials is needed that integrates the lessons learned through laboratory, translational, and clinical studies in recent years.


Employing tripodal carboxylate ligand to construct Co(II) coordination networks modulated by N-donor ligands: syntheses, structures and magnetic properties.
Hydrothermal reaction of a tripodal bridging ligand, 5-(4-carboxyphenoxy)isophthalic acid (H3cpia) with cobalt salts modulated by N-donor neutral ligands leads to the formation of six novel coordination networks formulated as {[Co(1.5)(cpia)(o-bix)](H2O)(1.5)}n (1), {[Co2(cpia)(μ-OH)(m-bix)]H2O}n (2), {[Co(1.5)(cpia)(m-bix)]}n (3), {[Co(1.5)(cpia)(p-bix)(0.5)(H2O)]H2O}n (4), {[Co(2.5)(cpia)(Hcpia)(4,4'-bpy)(2.5)](H2O)3}n (5), and {[Co3(cpia)2(bpp)2]H2O}n (6). Compound 1 exhibits a two-dimensional, (3,8)-connected layered architecture composed of trinuclear cobalt clusters. Compound 2 possesses a three-dimensional dense framework with (3,8)-connected tfz-d topology built from butterfly-shaped tetranuclear Co4(μ3-OH)2(6+) clusters. Similar to compound 1, trinuclear Co clusters act as secondary building units to construct the final 2D layered structure modulated by m-bix and bpp ligands in compounds 3 and 6. In compound 4, trinuclear Co clusters connected by cpia(3-) anions give rise to two-dimensional layers, which are further pillared by p-bix ligands to the three-dimensional framework. Compound 5 features a 2D, (3,4,6)-connected molecular network assembled from alternate binuclear and mononuclear Co building blocks. The magnetic investigation indicates that strong antiferromagnetic interactions between cobalt ions are dominant in compounds 2 and 6.


Using discrete choice experiment to elicit doctors' preferences for the report card design of diabetes care in Taiwan - a pilot study.
Little is known about doctors' preferences regarding public report card design. Taiwan just announced the first diabetes report card on April 2008. The aim of this study was to investigate the Diabetes Mellitus (DM) providers' preferences towards four report card attributes: update frequency, risk adjustment, content information and display format. A discrete choice questionnaire was mailed to hospital and primary care doctors in the northern part of Taiwan, with 221 study targets. The response rate was 29%. Using random effect logistic regression, doctors' preference attribute rankings were risk adjustment for patients (44.7%), content information (25.2%), display format (18.3%) and update frequency (11.8%). One-year update frequency, risk adjustment, detailed scores of technical quality and interpersonal quality and bar chart display were the most important items noted in our survey. This is the first study to investigate provider's preferences for a diabetes report card. It enables the policy maker to clearly see the implication of trade-offs between different choices when designing a report card that doctors will like. Our findings suggest that doctors do not favour the 'less is more' principle, or the higher frequency of updates that patients may prefer. Rather, our findings suggest that risk adjustment and more information content are the most important factors for doctors. Future studies should use discrete choice experiment on different aspects of report card design, such as vulnerable patients or health care administrations.


Ultra fast trypsin digestion of proteins by high intensity focused ultrasound.
Proteolytic digestion of proteins in seconds under an ultrasonic field provided by high-intensity focused ultrasound (HIFU) has been achieved. Successful in-solution and in-gel tryptic digestion of proteins in 60 s or less was demonstrated by either MALDI-TOF mass spectrometry or liquid chromatography-electrospray ion trap mass spectrometry (RP-HPLC-ESI-IT-MS/MS). The efficiency of this new procedure for protein digestion compared favorably with those attained using conventional overnight incubation methods. The performance of the method was also demonstrated by the specific identification of three proteins in a whole proteome in less than 1 h. The method greatly reduces the time needed for protein digestion, is of easy implementation, environmental friendly, and economic. Adaptation of this method to on-line procedures and robotic platforms could have promising applications in the proteomics field.


Accuracy and Tuning of Flow Parsing for Visual Perception of Object Motion During Self-Motion.
How do we perceive object motion during self-motion using visual information alone? Previous studies have reported that the visual system can use optic flow to identify and globally subtract the retinal motion component resulting from self-motion to recover scene-relative object motion, a process called flow parsing. In this article, we developed a retinal motion nulling method to directly measure and quantify the magnitude of flow parsing (i.e., flow parsing gain) in various scenarios to examine the accuracy and tuning of flow parsing for the visual perception of object motion during self-motion. We found that flow parsing gains were below unity for all displays in all experiments; and that increasing self-motion and object motion speed did not alter flow parsing gain. We conclude that visual information alone is not sufficient for the accurate perception of scene-relative motion during self-motion. Although flow parsing performs global subtraction, its accuracy also depends on local motion information in the retinal vicinity of the moving object. Furthermore, the flow parsing gain was constant across common self-motion or object motion speeds. These results can be used to inform and validate computational models of flow parsing.


Managed care, ethics, and academic health centers: maximizing potential, minimizing drawbacks.
As academic health centers (AHCs) enter into various forms of managed care, they face a number of managed care's well-known ethical issues: utilization controls and loss of clinical autonomy; incentives and conflicts of interest; strained collegial relations; and questionable origin and validity of practice guidelines. Also, special issues arise regarding teaching relationships, faculty structure, and standards of care. Although the ethical hazards of current forms of managed care are quite pervasive and arise largely through short-sighted cost controls, emerging forms of integration and payer-level capitation permit considerably greater flexibility to provide care that is coherent, comprehensive, even creative. AHCs, as integrated multispecialty group practices, may be in a surprisingly good position to explore the best possibilities of global capitation while minimizing the ethical drawbacks of current forms of managed care.


The role of human rights and personal dignity in the rehabilitation of chronic psychiatric patients. A rural therapeutic community in Yanbian, Jilin.
In 1983 the branch hospital of the Yanbian Community Psychiatric Hospital, a rural residential centre for 120 chronic psychiatric patients who have no means of financial support, adopted a new treatment philosophy that emphasised psychosocial rehabilitation and made the protection of patients' human rights and respect for their personal dignity the central organising principles for the hospital's treatment programme. From being a boarding facility for chronic psychiatric patients, the hospital became a thriving community. Comparison of the status of the 81 continuously resident schizophrenic patients before and after the policy change showed that (a) the proportion who actively participated in agricultural labour increased from 10% in 1982 to 38% in 1990; (b) the proportion who worked at non-agricultural jobs increased from 7% in 1982 to 22% in 1990; and (c) their mean yearly income increased from 1.67 Rmb in 1982 to 246.70 Rmb in 1990. Moreover, the number of successful suicides among all the patients in the hospital dropped from 13 in 1975-1982 to 1 in 1983-1990. We conclude that the success of psychiatric rehabilitation programmes depends on the extent to which they address the core issues of personal dignity and basic human rights.


Spontaneous resolution of a cystic hygroma in a fetus with Turner syndrome.
The prenatal detection of a cystic hygroma (CH) in a fetus with a 45,X karyotype is described. The cystic hygroma underwent spontaneous resolution and a healthy baby with Turner syndrome was subsequently born. The implications for genetic counselling are discussed.


Effects of intermittent administration of pamidronate on the mechanical properties of canine cortical and trabecular bone.
The mechanical properties of cortical and trabecular bones from beagles treated with the bisphosphonate pamidronate (administered intermittently 1 week every month for 3 months, at a dosage of 0.45 mumol/kg/day) were assessed. The mechanical properties of cortical bone were measured by four-point bending tests on femoral quadrants, in order to measure their elastic modulus and ultimate stress. The structural properties of whole tibias were measured in torsion to determine the torsional stiffness and failure torque. The elastic modulus and maximum compressive stress of the trabecular bone samples were measured by compression tests of trabecular cores. Intermittent treatment with pamidronate did not change the pattern of mechanical properties that occurs naturally around the femur or the torsional stiffness and failure torque of the tibias. By contrast, pamidronate did significantly increase the modulus of elasticity (by 19%) and maximum compressive stress (by 33%) of vertically aligned cylindrical trabecular specimens taken from the vertebrae of the beagles.


Changing the face of London's healthcare.
Europe's newest hospital, and the UK's largest ever PPP-funded and operated healthcare facility--the 650 pounds sterling million Royal London in Whitechapel--opened its doors on 1 March--following years of hard work and planning which has seen doctors and nurses involved throughout, working under the guidance of a 30-strong Barts and the London NHS Trust New Hospitals Programme team to create an optimal healing environment. HEJ editor, Jonathan Baillie, discussed with New Hospitals Programme director, Matthew Tulley, the challenges of overseeing the design, planning, and construction, of the 'new' Royal London as part of a wider 1 billion pounds sterling PFI project to transform and modernise facilities at both the Barts and the Royal London sites, and of smoothly moving over 3,000 staff and patients from old to new accommodation.


YAP regulates PD-L1 expression in human NSCLC cells.
Programmed death-ligand 1 (PD-L1) is a membrane protein on tumor cells that binds to the PD-1 receptor expressed on immune cells, leading to the immune escape of tumor cells. Yes-associated protein (YAP) is a main effector of the Hippo/YAP signaling pathway, which plays important roles in cancer development. Here we show that YAP regulates PD-L1 expression in human non-small cell lung cancer (NSCLC) cells. First, we investigated YAP and PD-L1 expression at the protein level in 142 NSCLC samples and 15 normal lung samples. In tumor tissue, immunohistochemistry showed positive staining for YAP and PD-L1, which correlated significantly (n = 142, r = 0.514, P < 0.001). Second, in cell lines that express high levels of PD-L1 (H460, SKLU-1, and H1299), the ratio of p-YAP/YAP was lower and GTIIC reporter activity of the Hippo pathway was higher than those in three cell lines expressing low levels of PD-L1 (A549, H2030, and PC9) (P < 0.05). Third, in the same three cell lines, inhibition of YAP by two small interfering RNAs (siRNAs) decreased the mRNA and protein level of PD-L1 (P < 0.05). Fourth, forced overexpression of the YAP gene rescued the PD-L1 mRNA and protein level after siRNA knockdown targeting 3'UTR of the endogenous YAP gene. Finally, chromatin immunoprecipitation (ChIP) assays using a YAP-specific monoclonal antibody resulted in the precipitation of PD-L1 enhancer region encompassing two putative TEAD binding sites. Our results indicate that YAP regulates the transcription of PD-L1 in NSCLC.


[Stem-cell engraftment as delayed therapy to repair organophosphate-induced brain damage].
Amongst organophosphate compounds, both pesticides and warfare neurotoxics are probably the most representative. These compounds are irreversible acetylcholinesterase inhibitors. Usual clinical signs observed after acute poisoning are mainly respiratory distress, convulsions and seizures. Following acute poisoning, an emergency treatment must be provided as soon as possible (maximum delay of 1 hour post-poisoning), to prevent irreversible brain damage and patient death. At the present time, there is no efficient delayed treatment which could be provided if this 1 hour latency is overpassed. However, neurogenesis by stem cell engraftment, eventually complemented by gene therapy strategy, could be a potential therapeutic approach to repair organophosphate-induced brain damage. Main stem cell engraftement strategies successfully used for brain damage of various origins are reviewed in this Article.


Prevalence, psychological impact, and risk factors of erectile dysfunction in patients with Peyronie's disease: a retrospective analysis of 309 cases.
Peyronie's disease (PD) is a chronic inflammatory disease involving the tunica albuginea of the penis. Erectile dysfunction (ED) is a possible invalidating symptom of PD. The aim of this study was to evaluate the prevalence, psychological impact, and risk factors of ED in patients with PD. The study was conducted by carrying out a retrospective analysis of the clinical records of 309 patients with PD who visited our andrology clinic. All patients underwent the following tests: body mass index, common blood tests and hormone assays, questionnaire for erectile function assessment, dynamic penile color Doppler ultrasonography, imaging of the penis at maximum erection with photographic poses according to Kelâmi, psychosexual impact evaluation with PD Questionnaire (symptom bother score), evaluation of depression symptoms with the Patient Health Questionnaire-9, and evaluation of the intensity of penile pain with the pain intensity numeric rating scale. ED was observed in 37.5% of the cases. We divided the cases into two groups: group A (PD + ED), 116 cases, and group B (PD without ED), 193 cases. After multivariate analysis, we concluded that the following comorbidities are independent risk factors for ED: dyslipidemia, obesity, chronic prostatitis, benign prostatic hyperplasia, and autoimmune diseases. A depressive disorder was observed in 62.4%, and it was more frequent in patients with PD + ED (91.37% versus 45.07% group B). Sexual bother was greater in group A compared with group B (9.7 versus 7.6). Intensities of depressive symptoms and sexual bother were significantly higher compared with cases with no curvature when the bend angle was ≥30°. Our study confirms that an integrated psychological support with medical treatment is needed in patients with PD.


Expression of miR-625 and Fas in cervical vertebral cartilage endplate.
The aim of the present study was to assess miR-625 and Fas expression in normal and degenerative cervical cartilage endplate (CEP) tissues. Following biof-informatics analysis, the Fas gene was predicted to be one of the targets of miR-625. Quantitative PCR (qPCR) and western blotting were used to detect miR-625 and Fas expression in normal and degenerative CEP. A luciferase reporter assay was used to identify whether miR-625 could directly target the 3' untranslated region (3'-UTR) of Fas. Lentiviral overexpression and/or inhibition vectors of miR-625 (pre-miR-625)/antigomiR-625 were constructed to determine whether overexpression or inhibition of miR-625 could affect Fas and B-cell lymphoma 2 (Bcl-2) expression in cartilaginous endplate cells (CECs) and tissues. qPCR analysis demonstrated that miR-625 expression in degenerative CEP was significantly lower than in normal CEP tissue, while the production of Fas in degenerated CEP was significantly higher. Results from western blotting also showed a significant increase in Fas expression in degenerative CEP. miR-625 can bind directly to the 3'-UTR of the Fas gene. However, this inhibition was attenuated by a target mutation in the miR-625-binding site of the 3'-UTR of Fas mRNA. In addition, following transfection of CECs with pre-miR-625 and antigomiR-625, expression of Fas significantly decreased and increased, respectively, and Bcl-2 expression was upregulated and downregulated, respectively. Upregulation of miR-625 can inhibit Fas expression and further affect Bcl-2 expression in CEP degeneration, suggesting that miR-625-mediated inhibition of the Fas gene is important in cervical degeneration.


Spiritual development of nursing students: developing competence to provide spiritual care to patients at the end of life.
Nurses spend more time with their patients than do other health care workers. Therefore, the spiritual needs of patients must be recognized as a domain of nursing care. Holism cannot exist without consideration of the spiritual aspects that create individuality and give meaning to people's lives. The purpose of this article is to provide nursing faculty with tools that may be used to develop spiritually knowledgeable nursing students who can overcome barriers to providing spiritual care to end-of-life patients. Our students were required to complete care maps to ensure they are prepared for patient care at the end of life. In this article, we present tools that faculty and students may use to complete the spiritual concept in care mapping. The literature on spirituality is reviewed, use of care mapping in nursing curricula is described, and our teaching approach to develop nursing students who are skilled at providing spiritual care is explained. Three case studies and care maps created by former students are also presented to demonstrate examples of spiritual competence.


Gillies Lecture: ocular motility in a time of paradigm shift.
Abstract Recent progress in understanding of the structure and function of extraocular muscles, and our ability to image them clinically, allows prediction of revolutionary progress in diagnosis and treatment of strabismus in the coming decades. This perspective memorializes a lecture given in honour of Dr William Gillies, who has for decades been the paternal leader of strabismology in southern Australia.


A perspective on diuretic resistance in chronic congestive heart failure.
Chronic congestive heart failure (CHF) is a complex disorder characterized by inability of the heart to keep up the demands on it, followed by the progressive pump failure and fluid accumulation. Although the loop diuretics are widely used in heart failure (HF) patients, both pharmacodynamic and pharmacokinetic alterations are thought to be responsible for diuretic resistance in these patients. Strategies to overcome diuretic resistance include sodium intake restriction, changes in diuretic dose and route of administration and sequential nephron diuretic therapy. In this review, we discuss the definition, prevalence, mechanism of development and management strategies of diuretic resistance in HF patients.


DNA-SeAl: Sensitivity Levels to Optimize the Performance of Privacy-Preserving DNA Alignment.
The advent of next-generation sequencing (NGS) machines made DNA sequencing cheaper, but also put pressure on the genomic life-cycle, which includes aligning millions of short DNA sequences, called reads, to a reference genome. On the performance side, efficient algorithms have been developed, and parallelized on public clouds. On the privacy side, since genomic data are utterly sensitive, several cryptographic mechanisms have been proposed to align reads more securely than the former, but with a lower performance. This paper presents DNA-SeAl a novel contribution to improving the privacy × performance product in current genomic workflows. First, building on recent works that argue that genomic data needs to be treated according to a threat-risk analysis, we introduce a multi-level sensitivity classification of genomic variations designed to prevent the amplification of possible privacy attacks. We show that the usage of sensitivity levels reduces future re-identification risks, and that their partitioning helps prevent linkage attacks. Second, after extending this classification to reads, we show how to align and store reads using different security levels. To do so, DNA-SeAl extends a recent reads filter to classify unaligned reads into sensitivity levels, and adapts existing alignment algorithms to the reads sensitivity. We show that using DNA-SeAl allows high performance gains whilst enforcing high privacy levels in hybrid cloud environments.


Properties of antibodies to thymine glycol, a product of the radiolysis of DNA.
Anti-thymine glycol antibodies were elicited by immunizing rabbits with thymine glycol monophosphate (TMP-glycol) conjugated by carbodiimide to BSA. The antibodies produced are specific for thymine glycol as measured by immunoprecipitation of TMP-glycol-RSA conjugates and hapten inhibition of reactivity with OsO4-treated DNA in an enzyme immunoassay. Using the enzyme immunoassay, the antibody is capable of detecting femtomole and picomole levels of thymine glycol in direct and competitive assays, respectively. This immunochemical assay is potentially suitable for measuring the production and repair of thymine glycol damage in cellular DNA.


Fiber-reinforced fixed dental prosthesis to replace missing anterior teeth: case reports.
The direct fiber-reinforced fixed dental prosthesis (FRFDP) is a minimally invasive approach to replacing missing anterior teeth, offering good survival rates and sound mechanical properties. This report describes the placement and 12-month follow-up results of FRFDPs used to replace missing anterior teeth in 2 patients. In the first case, the extraction of the maxillary right central incisor was related to trauma. In the second case, severe periodontal disease necessitated the extraction of the mandibular left central incisor. Based on the biological and financial considerations of the patients and the need to obtain clinical resolution in a short time, placement of an FRFDP was proposed in each case. Each patient was informed of the benefits and limitations of an FRFDP and accepted the treatment plan. In each case, an FRFDP was placed immediately after the surgical extraction of the incisor, using the extracted tooth as a pontic. The results observed at the 12-month follow-up examinations of the patients suggest that the FRFDP is a viable, esthetic, and functional option to replace missing anterior teeth.


[Surgical treatment of pancreatic cancer-What is new?]
The incidence of pancreatic ductal adenocarcinoma is continuously increasing and will become the second leading cause of cancer-related death in Europe and the USA by 2030. With a 5-year overall survival rate of less than 10% the prognosis remains poor. So far surgical tumor resection remains the only curative treatment option, which is now partially supported by multimodal neoadjuvant and adjuvant therapy concepts. Due to the aggressive tumor biology patients with advanced pancreatic cancer in particular can profit from these multimodal therapy concepts. Additionally, in recent years surgical treatment was optimized, the criteria for tumor resectablity were defined and minimally invasive surgery was widely introduced. This review article summarizes the newest developments and the new German S3 guidelines concerning surgery of pancreatic cancer.


Epigenetics of sex determination and gonadogenesis.
Epigenetics is commonly defined as the study of heritable changes in gene function that cannot be explained by changes in DNA sequence. The three major epigenetic mechanisms for gene expression regulation include DNA methylation, histone modifications, and non-coding RNAs. Epigenetic mechanisms provide organisms with the ability to integrate genomic and environmental information to modify the activity of their genes for generating a particular phenotype. During development, cells differentiate, acquire, and maintain identity through changes in gene expression. This is crucial for sex determination and differentiation, which are among the most important developmental processes for the proper functioning and perpetuation of species. This review summarizes studies showing how epigenetic regulatory mechanisms contribute to sex determination and reproductive organ formation in plants, invertebrates, and vertebrates. Further progress will be made by integrating several approaches, including genomics and Next Generation Sequencing to create epigenetic maps related to different aspects of sex determination and gonadogenesis. Epigenetics will also contribute to understand the etiology of several disorders of sexual development. It also might play a significant role in the control of reproduction in animal farm production and will aid in recognizing the environmental versus genetic influences on sex determination of sensitive species in a global change scenario.


Increasing length of an estradiol and progesterone timed artificial insemination protocol decreases pregnancy losses in lactating dairy cows.
Our hypothesis was that increasing the length of an estradiol and progesterone (P4) timed artificial insemination (TAI) protocol would improve pregnancy per artificial insemination (P/AI). Lactating Holstein cows (n=759) yielding 31 ± 0.30 kg of milk/d with a detectable corpus luteum (CL) at d -11 were randomly assigned to receive TAI (d 0) following 1 of 2 treatments: (8d) d -10 = controlled internal drug release (CIDR) and 2.0mg of estradiol benzoate, d -3 = PGF2α(25mg of dinoprost tromethamine), d -2 = CIDR removal and 1.0mg of estradiol cypionate, d 0 = TAI; or (9 d) d -11 = CIDR and estradiol benzoate, d -4 = PGF2α, d -2 CIDR removal and estradiol cypionate, d 0 TAI. Cows were considered to have their estrous cycle synchronized in response to the protocol by the absence of a CL at artificial insemination (d 0) and presence of a CL on d 7. Pregnancy diagnoses were performed on d 32 and 60. The ovulatory follicle diameter at TAI (d 0) did not differ between treatments (14.7 ± 0.39 vs. 15.0 ± 0.40 mm for 8 and 9 d, respectively). The 9 d cows tended to have greater P4 concentrations on d 7 in synchronized cows (3.14 ± 0.18 ng/mL) than the 8d cows (3.05 ± 0.18 ng/mL). Although the P/AI at d 32 [45 (175/385) vs. 43.9% (166/374) for 8d and 9 d, respectively] and 60 [38.1 (150/385) vs. 40.4% (154/374) for 8d and 9 d, respectively] was not different, the 9 d cows had lower pregnancy losses [7.6% (12/166)] than 8d cows [14.7% (25/175)]. The cows in the 9 d program were more likely to be detected in estrus [72.0% (269/374)] compared with 8d cows [62% (240/385)]. Expression of estrus improved synchronization [97.4 (489/501) vs. 81% (202/248)], P4 concentrations at d 7 (3.22 ± 0.16 vs. 2.77 ± 0.17 ng/mL), P/AI at d 32 [51.2 (252/489) vs. 39.4% (81/202)], P/AI at d 60 [46.3 (230/489) vs. 31.1% (66/202)], and decreased pregnancy loss [9.3 (22/252) vs. 19.8% (15/81)] compared with cows that did not show estrus, respectively. Cows not detected in estrus with small (<11 mm) or large follicles (>17 mm) had greater pregnancy loss; however, in cows detected in estrus, no effect of follicle diameter on pregnancy loss was observed. In conclusion, increasing the length of the protocol for TAI increased the percentage of cows detected in estrus and decreased pregnancy loss.


[A case of gastric carcinoid tumor with multiple liver metastasis].
Carcinoid tumors originate from the enterochromaffin cells in the gastrointestinal mucosa. Gastric carcinoids are rare and usually well-differentiated tumors with variable prognosis. Three types of gastric carcinoids have been recognized. Type I is associated with type A chronic atrophic gastritis, achlorhydria and pernicious anemia. Type II is associated with Zollinger-Ellison syndrome and multiple endocrine neoplasia type I. Type III appears as a large malignant neoplasm and has no association with atrophic gastritis or hypergastrinemia. We experienced a rare case of gastric carcinoid tumor type III with multiple liver metastasis. Thus, we report here this case with a review of the literature.


Suitability of aromatic plants for phytoremediation of heavy metal contaminated areas: a review.
This review briefly elucidates the research undertaken and benefits of using aromatic plants for remediation of heavy metal polluted sites. A sustainable approach to mitigate heavy metal contamination of environment is need of the hour. Phytoremediation has emerged to be one of the most preferable choices for combating the metal pollution problem. Aromatic plants can be used for remediation of contaminated sites as they are non-food crops thus minimizing the risk of food chain contamination. Most promising aromatic plants for phytoremediation of heavy metal contaminated sites have been identified from families - Poaceae, Lamiaceae, Asteraceae, and Geraniaceae. They act as potential phytostabilisers, hyper accumulators, bio-monitors, and facultative metallophytes. Being high value economic crops, monetary benefits can be obtained by growing them in tainted areas instead of food crops. It has been observed that heavy metal stress enhances the essential oil percentage of certain aromatic crops. Research conducted on some major aromatic plants in this context has been highlighted in the present review which suggests that aromatic plants hold a great potential for phytoremediation. It has been reported that essential oil from aromatic crops is not contaminated by heavy metals significantly. Thus, aromatic plants are emerging as an ideal candidate for phytoremediation. Highlights • Aromatic plants hold a great potential for phytoremediation of heavy metal contaminated sites. • Being high value economic crops, monetary benefits can be obtained by growing them in contaminated areas instead of food crops. • Research done on some major aromatic plants in this context has been highlighted in the present review.


Structural insights into the coordination and extraction of Pb(II) by disulfonamide ligands derived from o-phenylenediamine.
The o-phenylenediamine-derived disulfonamide ligands 1 and 2 complex and efficiently extract Pb(II) from water into 1,2-dichloroethane via ion-exchange, in combination with 2,2'-bipyridine (97.5% and 95.0%, respectively, for 1:1 ligand-to-Pb ratios). The corresponding Pb(II)-sulfonamido binary complexes of ligands 1 and 2 (3 and 4, respectively), and ternary complexes with 2,2'-bipyridine (5 and 6, respectively), were isolated and characterized. (1)H NMR spectra of the organic phases after extraction show the formation of ternary Pb-sulfonamido-bipy complexes. X-ray characterization of 3, 4, and the ternary complex 5 consistently demonstrates four primary coordination sites and a stereochemically active lone pair on Pb. The X-ray structure of 3 shows a pseudo trigonal bipyramidal configuration on Pb, with the lone pair occupying one of the equatorial sites, and the formation of an unusual "hemidirected" coordination polymer via axial S=O-Pb coordination. The same axial S=O-Pb coordination pattern with two DMSO molecules is observed in the structure of 4.[2(CH(3))(2)SO)], thus rationalizing the high solubility of the binary complexes in strongly coordinating solvents. In contrast, the X-ray structure of the ternary complex 5 reveals a distorted four-coordinate configuration with only weak S=O-Pb coordination leading to dimer formation, thus explaining its higher solubility in weakly coordinating solvents. FT-IR spectroscopy confirms the X-ray data, since the ligand nu(S)(=)(O) stretching frequencies shift to lower values in the binary Pb(II)-sulfonamido complexes and are again altered upon formation of the ternary Pb(II)-sulfonamido-bipy complexes, as would be expected for 2,2'-bipy complexation and hindered S=O-Pb coordination.


Primary Buttonhole Mastopexy and Nipple-Sparing Mastectomy: A Preliminary Report.
Patients undergoing nipple-sparing mastectomy and immediate-implant based reconstruction occasionally require a mastopexy based on their breast size and degree of ptosis. Previous reports have shown the feasibility of mastopexy-nipple-sparing mastectomy in selected patients to raise the nipple up to 5 cm. Major mastopexy with nipple transposition more than 6 cm in conjunction with nipple-sparing mastectomy for therapeutic indications has not been described. The authors review their experience with primary buttonhole mastopexy performed in conjunction with nipple-sparing mastectomy. Between 2008 and 2014, 16 patients (32 breasts) underwent bilateral primary mastopexy and nipple-sparing mastectomy with immediate staged implant-based reconstruction. The Passot buttonhole technique was used for the mastopexy in all patients, raising the nipple from 7 to 12 cm. Tumor-related data, risk factors, breast size, degree of ptosis, expander size, fill volume, selection criteria, and complications are discussed. The average follow-up period was 33 months (range, 14 to 80 months). There were no tumor recurrences, and all patients completed their reconstruction. Two patients required removal of the expander and delayed reconstruction because of infection and implant exposure due to nipple-areola loss. The reasons for nipple-areola loss and technical modifications to enhance skin viability by retaining a thin layer of subareolar breast tissue for removal during the second-stage implant exchange are discussed. Primary mastopexy using the buttonhole technique performed together with nipple-sparing mastectomy is a safe procedure with predictable results in patients with very large or ptotic breasts requiring lifts greater than 6 cm. The success of the combined procedure depends on preserving a thin layer of subareolar breast tissue and removing it at the time of implant exchange.


Glucocorticoids enhance Th17/Th1 imbalance and signal transducer and activator of transcription 3 expression in systemic lupus erythematosus patients.
To investigate the relative amounts of Th17 and Th1 cells present in SLE patients and the possible effects of treatments or disease features on these populations. Peripheral blood mononuclear cells were collected from 75 SLE patients and 19 healthy controls and the proportion of Th17 and Th1 populations were assessed by flow cytometry measuring the amount of IL-17 and IFN-γ-producing cells. Gene expression of signal transducers and activators of transcription 3 (STAT3), STAT4, IL-6R and IL-12R were determined in 30 patients and 8 healthy individuals by real-time RT-PCR. Data were related to clinical and immunological parameters and to the treatment followed during the past 3 months. Th17 cells and the Th17/Th1 ratio were significantly increased in SLE patients treated with glucocorticoids compared with healthy individuals, untreated patients or those under other treatments. No association was detected with clinical parameters, but patients with anti-ENA antibodies also displayed increased Th17 responses. Disease activity (SLEDAI) is associated with the Th17/Th1 index only in glucocorticoid-treated patients. In line with these results, gene expression of STAT3 and IL-6R was up-regulated in patients taking these drugs. Accordingly, we found a positive correlation between the Th17/Th1 ratio and STAT3 levels. The present work provides the first evidence that aberrant Th17/Th1 balance in SLE is linked to the use of glucocorticoids and suggests that the up-regulatory effect of these drugs on the Th17 population could be associated with their ability to increase STAT3 and IL-6R expression. Additionally, anti-ENA positivity could represent a potential biomarker for Th17 bias.


Long-term survival of porcelain laminate veneers using two preparation designs: a retrospective study.
This study evaluated the long-term survival of anterior porcelain laminate veneers placed with and without incisal porcelain coverage. Two prosthodontists in a private dental practice placed 110 labial feldspathic porcelain veneers in 50 patients; 46 veneers were provided with incisal porcelain coverage, and 64 were not. The veneers were evaluated retrospectively from case records for up to 7 years (mean 4 years). At 5, 6, and 7 years, the cumulative survival estimates were 95.8% for veneers with incisal porcelain coverage and 85.5% for those without incisal coverage. The difference was not statistically significant. Six of the nine failures occurred from porcelain fracture in the veneers without incisal coverage. Although there was a trend for better long-term survival of the veneers with incisal porcelain coverage, this finding was not statistically significant.


Neuroprotective effect of cocoa flavonoids on in vitro oxidative stress.
Cocoa is a rich source of flavonoids that, among other functions, can act as antioxidants. In living systems, the production of reactive oxygen species (ROS) activate an array of intracellular cascades, including mitogen-activated protein kinases (MAPK), that are closely associated with cell death or survival pathways. To ascertain the role of a cocoa extract and its main flavonoid, (-)-epicatechin, in an in vitro model of oxidative stress induced in a neuronal cell line. We analyzed ROS production by fluorometry (dichlorofluorescein assay), and activation of MAPK pathways including extracellular signal-regulated kinases 1/2 (ERK 1/2), c-Jun N-terminal kinase (JNK), and p-38, by Western blot analysis. Cells incubated with cocoa extract or (-)-epicatechin, reduced ROS production in a dose-dependent manner, reaching 35% inhibition. pJNK and p38, involved in apoptosis, were down-modulated by cocoa extract and (-)-epicatechin with p38 inhibition reaching up to 70%. Our results show that cocoa extract and (-)-epicatechin may exert a neuroprotective action by reducing ROS production and modulating MAPK activation.


Neuroprotective Effect and Mechanism of Thiazolidinedione on Dopaminergic Neurons In Vivo and In Vitro in Parkinson's Disease.
The aim of the present study was to gain insight into the neuroprotection effects and mechanism of thiazolidinedione pioglitazone in both in vitro and in vivo MPP+/MPTP induced PD models. In vivo experimental results showed that oral treatment of pioglitazone resulted in significant improvements in behavior symptoms damaged by MPTP and increase in the survival of TH positive neurons in the pioglitazone intervention groups. In addition, oral treatment of pioglitazone increased the expression of peroxisome proliferator-activated receptor-γ coactivator of 1α (PGC-1α) and increased the number of mitochondria, along with an observed improvement in mitochondrial ultrastructure. From in vitro studies, 2,4-thiazolidinedione resulted in increased levels of molecules regulated function of mitochondria, including PGC-1α, nuclear respiratory factor 1 (NRF1), NRF2, and mitochondria fusion 2 (Mfn2), and inhibited mitochondria fission 1 (Fis1). We show that protein levels of Bcl-2 and ERK were reduced in the MPP+-treated group compared with the control group. This effect was observed to be reversed upon treatment with 2,4-thiazolidinedione, as Bcl-2 and ERK expression levels were increased. We also observed that levels of the apoptotic protein Bax showed opposite changes compared to Bcl-2 and ERK levels. The results from this study confirm that pioglitazone/2,4-thiazolidinedione is able to activate PGC-1α and prevent damage of dopaminergic neurons and restore mitochondria ultrastructure through the regulation of mitochondria function.


[Relationship between overnight urinary sodium to potassium ratio and the risk of cardiovascular disease].
Objective: To explore the relationship between overnight urinary sodium to potassium ratio and the risk of cardiovascular disease (CVD). Methods: A subsample of 10 percent of the participants (35-59 years old) from the People's Republic of China-United States Collaborative Study of Cardiovascular and Cardiopulmonary Epidemiology (prospective survey) were used. Three consecutive overnight urine samples were collected in the autumn of 1983-1984 and the spring in 1985-1986, respectively. Urinary sodium and potassium were detected and calculated for 8 hours excretion. The occurrences of cardiovascular events were recorded in 2 years interval from 1987-1988 until December 31, 2005. Participants were divided into first ratio group, second ratio group, and third ratio group based on the tertiles of sodium to potassium ratio. Cox proportional hazard regression model was used to determine the relationship between sodium to potassium ratio and risk of CVD. In addition, participants were divided into 2 subgroups by the median of overnight urinary sodium and potassium, and then combined each other for 4 subgroups including low sodium-low potassium group, low sodium-high potassium group, high sodium-low potassium group, and high sodium-high potassium group, to explore the relationship between different sodium-potassium combinations and the risk of CVD. Results: A total of 954 participants were included in the final analysis, of whom 459 (48.1%) were males. There were 318 cases in the first, second and third ratio group, respectively. There were 347 cases in low sodium-low potassium group and high sodium-high potassium group, and 130 cases in low sodium-high potassium group and high sodium-low potassium group. After a median follow-up of 18.6 (18.3, 19.3) years, cardiovascular events occurred in 81 participants, including 64 stroke and 20 coronary heart disease events. Multivariate analysis showed that comparing with the first ratio group, the hazard ratios (HR) in the second and the third ratio groups were 2.04 (95%CI 1.06-3.95, P=0.034) and 2.07 (95%CI 1.07-4.03, P=0.032), respectively. The CVD risk in low sodium-low potassium group was 24% higher than the low sodium-high potassium group (reference), but this result did not reach statistical significant level (P=0.685). The risks in high sodium-high potassium group (HR=3.32, 95%CI 1.26-8.76,P=0.015) and high sodium-low potassium (HR=3.04, 95%CI 1.05-8.83, P=0.041) group were both significantly increased. Conclusions: Overnight urinary sodium to potassium ratio is positively correlated with the risk of cardiovascular events. High urinary sodium plays a more important role for the increased risk of cardiovascular events than low potassium.


Simultaneous measurements of cardiac output by thermodilution, esophageal Doppler, and electrical impedance in anesthetized patients.
Simultaneous intraoperative measurements of cardiac output were obtained in nine patients with transesophageal Doppler, transthoracic impedance, and pulmonary artery thermodilution techniques to evaluate the utility of the noninvasive methods. Pairs of noninvasive and thermodilution measurements were obtained 25 times with transesophageal Doppler and 58 times with transthoracic impedance. Correlation of the noninvasive measurements with thermodilution was poor, with r = 0.43 for transthoracic impedance and r = .68 for transesophageal Doppler. The average difference between the noninvasive and the thermodilution values was -0.4 +/- 1.4 L/min (mean +/- SD) and -0.1 +/- 1.6 L/min for impedance and Doppler, respectively. Changes in cardiac output at sequential time points as measured by thermodilution were predicted with 95% confidence only when a change of >4 L/min was observed by transesophageal Doppler or >8 L/min was observed by transthoracic impedance. Therefore, it is concluded that neither noninvasive technique reliably estimated cardiac output as determined by thermodilution, and neither tracked trends.


Role of Kallistatin Treatment in Aging and Cancer by Modulating miR-34a and miR-21 Expression.
Kallistatin is an endogenous protein that regulates differential signaling pathways and a wide spectrum of biological activities via its two structural elements: an active site and a heparin-binding domain. Kallistatin via its heparin-binding site inhibits vascular inflammation and oxidative stress by antagonizing TNF-α-induced NADPH oxidase activity, NF-κB activation, and inflammatory gene expression in endothelial cells. Moreover, kallistatin via its active site inhibits microRNA-34a (miR-34a) synthesis and stimulates eNOS and SIRT1 expression in endothelial progenitor cells, whereas its heparin-binding site is crucial for blocking TNF-α-induced miR-21 expression and oxidative stress, thus reducing cellular senescence. By downregulating miR-34a and miR-21 expression, kallistatin treatment attenuates oxidative damage and aortic senescence in streptozotocin-induced diabetic mice and extends Caenorhabditis elegans lifespan under stress conditions. Likewise, kallistatin through the heparin-binding site inhibits TGF-β-induced miR-21 synthesis and oxidative stress in endothelial cells, resulting in inhibition of endothelial-mesenchymal transition, a process contributing to fibrosis and cancer. Furthermore, kallistatin's active site is essential for stimulating miR-34a and p53 expression and inhibiting the miR-21-Akt-Bcl-2 signaling pathway, thus inducing apoptosis in breast cancer cells. These findings reveal novel mechanisms of kallistatin in protection against senescence, aging, and cancer development by modulating miR-34a and miR-21 levels and inhibiting oxidative stress.


Respiratory syncytial virus infection of human alveolar epithelial cells enhances interferon regulatory factor 1 and interleukin-1beta-converting enzyme gene expression but does not cause apoptosis.
The induction kinetics of the transcriptional activities of interferon regulatory factor 1 (IRF-1), interleukin-1beta-converting enzyme (ICE), and CPP32 by respiratory syncytial virus (RSV) infection of human type II alveolar epithelial cells (A549 cells) were analyzed semiquantitatively by reverse transcriptase PCR. The appearance of ICE and CPP32 protein in cell lysate was examined by Western blotting analysis. The induction of apoptosis by RSV infection was examined by the appearance of DNA fragmentation detected by terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling. RSV moderately enhanced IRF-1 mRNA as early as 4 h after infection, and this enhancement lasted several hours. Following induction of the IRF-1 gene, ICE gene expression increased significantly, and an increase of ICE protein was observed in the RSV-infected cell lysate. These increments were observed in cells treated with live RSV but not in cells treated with inactivated RSV or control antigen. However, no infection-specific increase of CPP32 gene expression or the protein was observed. No nucleosomal fragmentation was observed in RSV-infected cells during the whole course of infection, despite the appearance of extensive cytopathic change and cell death. These observations suggest that RSV infection of human alveolar epithelial cells induces the ICE gene and its protein as a result of increased IRF-1 induction but that the increased ICE was insufficient to cause apoptosis in the RSV-infected cells. ICE might not be able to activate CPP32, which is thought to be the more important protease for apoptosis.


Identification, Quantitation, and Sensory Evaluation of Thiols in Bordeaux Red Wine with Characteristic Aging Bouquet.
Great Bordeaux red wines are known for their distinctive aging bouquet. However, the nature of volatile chemicals underpinning this sensory expression is not fully understood. This work investigated the empyreumatic aging bouquet of a collection of premium Bordeaux red wines using silver-ion (Ag+) solid-phase extraction, cryogenic heart-cutting multidimensional gas chromatography mass spectrometry/olfactometry, and comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry. In doing so, a substantial number of "meaty" odors were revealed. Three detected "meaty" notes were tentatively or unequivocally attributed to furan thiols. Among them, 2-methyltetrahydrofuran-3-thiol (1) with a pleasant "meaty" aroma was reported in wine for the first time. Its trans isomer (trans-1a) was resolved from its racemate by chemical modification, which confirmed its presence in wine. The odor detection threshold of trans-1a in the model wine was determined at 55 ng/L. Moreover, an additive effect between 1 and literature-known 2-methyl-3-furanthiol was observed. By a new ultra high-performance liquid chromatography quadrupole Orbitrap high-resolution mass spectrometry method, the concentration of trans-1a, in addition to those of 2-methyl-3-furanthiol and 2-furfuryl thiol, was measured in the wines at ng/L levels.


Access to Genetic Counselors in the Southern United States.
The expansion of genetic and genomic testing across medical specialties and the changing workforce demographics of certified genetic counselors (CGCs) have led to concerns of a workforce shortage. We assessed the number of genetic counselors working in the Southern United States-a rural and medically underserved region-using various online and professional resources. We identified 683 practicing genetic counselors across the Southern U.S. and 160 specializing in prenatal genetics. CGCs were concentrated in urban areas; counties with a CGC had a significantly higher proportion of minority residents and median household income than counties without a CGC. There is an average of 2.97 prenatal CGCs per 5000 high-risk births in the South. Alternative delivery models are needed to increase access to counseling services in the Southern U.S., particularly for low income households and those of high risk pregnancies. Increased provider education and patient educational materials can help facilitate informed decision-making in prenatal settings as genetic technologies gain a stronger foothold and bring value to medical practice.


Tetracycline nanoparticles as antibacterial and gene-silencing agents.
The spread of antibiotic-resistant bacteria and parasites calls for the development of new therapeutic strategies with could potentially reverse this trend. Here, a proposal is presented to exploit a sonochemical method to restore the antibiotic activity of tetracycline (TTCL) against resistant bacteria by converting the antibiotic into a nanoparticulate form. The demonstrated sonochemical method allows nanoscale TTCL assembly to be driven by supramolecular hydrogen bond formation, with no further modification to the antibiotic's chemical structure. It is shown that tetracycline nanoparticles (TTCL NPs) can act as antibacterial agents, both against TTCL sensitive and against resistant bacterial strains. Moreover, the synthesized antibiotic nanoparticles (NPs) can act as effective gene-silencing agents through the use of a TTCL repressor in Trypanosome brucei parasites. It is demonstrated that the NPs are nontoxic to human cells and T. brucei parasites and are able to release their monomer components in an active form in a manner that results in enhanced antimicrobial activity relative to a homogeneous solution of the precursor monomer. As the TTCL NPs are biocompatible and biodegradable, sonochemical formation of TTCL NPs represents a new promising approach for generation of pharmaceutically active nanomaterials.


Scanning electron microscopy of CV-1 and HeLa cells infected with herpes simplex viruses types 1 and 2.
Production of virus particles in CV-1 or HeLa BU-25 cells was investigated after their infection with several strains of herpes simplex virus (HSV) types 1 and 2, especially in relation to the association of virus particles with cells, the ratio of plaque forming units (PFU) to the whole number of virus particles and the morphological characteristics of cytopathic effect (CPE). Growth curves of the viruses differed according to the combination of cells and infecting virus strains. At 20 hr p.i., the number of cell-associated or cell-free viruses ranged from 5 X 10(8) to 5 X 10(9) PFU/35 mm dish or from 5 X 10(2) to 1.5 X 10(4) PFU/cell irrelevant of virus serotype or the morphology of CPE. In the case of CV-1 cells, the ratios of the number of the virus particles to PFU ranged from 100 to 640 and/or 18 to 110, respectively, depending on the CPE of rounding type or of fusion type. In case of CPE of fusion type, a higher rate of infectious particles was observed.


YcaO-mediated ATP-dependent peptidase activity in ribosomal peptide biosynthesis.
YcaO enzymes catalyze ATP-dependent post-translation modifications on peptides, including the installation of (ox/thi)azoline, thioamide and/or amidine moieties. Here we demonstrate that, in the biosynthesis of the bis-methyloxazolic alkaloid muscoride A, the YcaO enzyme MusD carries out both ATP-dependent cyclodehydration and peptide bond cleavage, which is a mechanism unprecedented for such a reaction. YcaO-catalyzed modifications are proposed to occur through a backbone O-phosphorylated intermediate, but this mechanism remains speculative. We report, to our knowedge, the first characterization of an acyl-phosphate species consistent with the proposed mechanism for backbone amide activation. The 3.1-Å-resolution cryogenic electron microscopy structure of MusD along with biochemical analysis allow identification of residues that enable peptide cleavage reaction. Bioinformatics analysis identifies other cyanobactin pathways that may deploy bifunctional YcaO enzymes. Our structural, mutational and mechanistic studies expand the scope of modifications catalyzed by YcaO proteins to include peptide hydrolysis and provide evidence for a unifying mechanism for the catalytically diverse outcomes.


Liquid chromatography-mass spectrometry method for determination of tetramethylpyrazine and its metabolite in dog plasma.
A liquid chromatography-mass spectrometry method is described for the determination of tetramethylpyrazine (TMP) and its active metabolite, 2-hydroxymethyl-3,5,6-trimethylpyrazine (HTMP) in dog plasma. This method involves a plasma clean-up step using protein precipitation procedure followed by LC separation and positive electrospray ionization mass spectrometry detection (ESI-MS). Chromatographic separation of the analytes was achieved on a C18 column using a mobile phase of methanol, water and acetic acid (50:50:0.6, v/v/v) at a flow rate of 1.0 ml/min. Selected ion monitoring (SIM) mode was used for analyte quantitation at m/z 137.2 for TMP, m/z 153.2 for HTMP and m/z 195.2 for caffeine. The linearity was obtained over the concentration ranges of 20-6000 ng/ml for TMP and 20-4000 ng/ml for HTMP and the lower limit of quantitation was 20 ng/ml for both analytes. For each level of QC samples, both inter- and intra-day precisions (R.S.D.) were <or=7.4% for TMP and <or=6.0% for HTMP, and accuracy (R.E.) was +/-6.0% for TMP and <or=3.5% for HTMP. The proposed LC-MS method was successfully applied to the pharmacokinetic studies of a TMP formulation preparation after oral administration to beagle dogs.


Elevated plasma FXII is associated with disease activity in ANCA-associated vasculitis: A Retrospective Cross-Sectional Study in Western China.
This study examined whether plasma FXII levels reflect disease activity in antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV). Plasma FXII levels were detected by ELISA in 127 patients with AAV, and their associations with disease activity and plasma myeloperoxidase (MPO)-ANCA titre were examined. Immunofluorescent co-staining of FXII and neutrophils was performed on the renal tissues of patients with AAV. MPO expression in renal biopsy tissues was determined by immunohistochemical staining. The association between plasma FXII levels and histological activity was assessed in 82 patients who underwent kidney biopsy. Plasma FXII levels were considerably increased in patients with clinically active AAV compared to those in clinical remission and healthy individuals. Plasma FXII levels correlated positively with creatinine (r = 0.377), CRP (r = 0.222), urine red blood cell (r = 0.203), serum MPO-ANCA titer (r = 0.353), white blood cell (r = 0.194), percentage of glomeruli with crescents (P = 0.001), capillary breaks (P = 0.001), interstitial inflammation (P < 0.001) and fibrinoid necrosis (p < 0.001) on kidney biopsy. The plasma FXII optimal cut-off value for evaluating AAV activity was 24.5 μg/mL (sensitivity = 0.81, specificity = 0.82, P = 0.0001), which was superior to that achieved using conventional serologic biomarkers. Co-expression of FXII and neutrophils was higher, with increased MPO expression, in renal tissue with pathologically active AAV than that observed in pathologically inactive tissues. In conclusion, elevated plasma FXII levels reflect AAV clinical and histologic activity, and can serve as markers of active AAV.


Essential debate.
With the election settled, the Obama administration has forged ahead with a draft rule on essential benefits and other provisions under the Affordable Care Act. Key players are still digesting the proposals, but HHS Secretary Kathleen Sebelius, left, says she's confident that the new rules-and an extended deadline related to exchanges-will give states what they need to implement the law.


Transient expression assays in grapevine: a step towards genetic improvement.
In the past few years, the usefulness of transient expression assays has continuously increased for the characterization of unknown gene function and metabolic pathways. In grapevine (Vitis vinifera L.), one of the most economically important fruit crops in the world, recent systematic sequencing projects produced many gene data sets that require detailed analysis. Due to their rapid nature, transient expression assays are well suited for large-scale genetic studies. Although genes and metabolic pathways of any species can be analysed by transient expression in model plants, a need for homologous systems has emerged to avoid the misinterpretation of results due to a foreign genetic background. Over the last 10 years, various protocols have thus been developed to apply this powerful technology to grapevine. Using cell suspension cultures, somatic embryos, leaves or whole plantlets, transient expression assays enabled the study of the function, regulation and subcellular localization of genes involved in specific metabolic pathways such as the biosynthesis of phenylpropanoids. Disease resistance genes that could be used for marker-assisted selection in conventional breeding or for stable transformation of elite cultivars have also been characterized. Additionally, transient expression assays have proved useful for shaping new tools for grapevine genetic improvement: synthetic promoters, silencing constructs, minimal linear cassettes or viral vectors. This review provides an update on the different tools (DNA constructs, reporter genes, vectors) and methods (Agrobacterium-mediated and direct gene transfer methods) available for transient gene expression in grapevine. The most representative results published thus far are then described.


Biomechanical, metabolic and cardiopulmonary responses of masters recreational runners during running at different speeds.
This study tested interactions between age and running speed on biomechanics, metabolic responses and cardiopulmonary responses. Three-hundred participants ran at preferred and standardized speeds. Age group (younger, masters [≥40 years]) by speed (self-selected 8.8 km/h, 11.2 km/h and 13.6 km/h) interactions were tested on main outcomes of sagittal kinematic, temporal spatial, metabolic and cardiopulmonary parameters. At all speeds, angular displacements of the ankle, pelvis and knee were less in masters than younger runners (Hedges g effect size range = 0.30-1.04; all p < 0.05). A significant age group by speed interaction existed for hip angular displacement (Wald χ2 = 10.753; p = 0.013). Masters runners ran at higher relative heart rates (p < 0.05) but at similar rates of oxygen use and energy expenditure. Masters runners used hip-dominant motion and step lengthening as running speed increased, but did not change centre of mass vertical displacement. This may increase mechanical stresses on tissues of the lower extremity in masters runners, especially hamstrings, hip joint and Achilles.


[Establishment and application of an analytical method for PINK1 gene exon copy number].
To establish a method for analyzing the PTEN-induced kinase 1 gene (PINK1) exon copy number and apply it to the analysis of PINK1 gene exon copy number variation (CNV) in patients with autosomal recessive early-onset Parkinsonism (AREP). Real-time PCR was used to analyze the exon copy number in 22 probands with AREP from unrelated Chinese Han families and 30 healthy controls. Copy numbers of exons 1-8 of the PINK1 gene were analyzed, and satisfactory reaction conditions and primers for exons of the PINK1 gene were obtained. No exon CNV in the PINK1 gene was detected in this group. An analytical method for PINK1 gene exon copy number was established. The exon CNV in the PINK1 gene was rare in Chinese patients with AREP.


Color vision testing by Farnsworth lantern and ability to identify approach-path signal colors.
It is presumed that pilots with abnormal color vision who pass the Farnsworth lantern test can distinguish aviation signal lights including the red and white lights of the Precision Approach Path Indicator (PAPI). The investigation of the FedEx Flight 1478 crash at Tallahassee airport in 2002 raises doubts about this presumption. It found a contributory factor was that the pilot had difficulty distinguishing the PAPI signals because of his deficient color vision (DCV) even though he had passed the Farnsworth lantern. This paper reports on the ability of subjects with DCV who pass the Farnsworth lantern to distinguish PAPI signal colors. Under a good visibility and a marginal visibility condition, 52 DCV subjects and 52 with normal color vision (NCV) named the colors of simulated PAPI signal lights observed in the dark. Subjects were tested with the Farnsworth lantern and a comprehensive battery of other color vision tests. The 10 DCV subjects who passed the Farnsworth lantern made significantly more errors naming PAPI signals than the NCV subjects and 80% made more errors than the worst performing NCV subject. Some confused the red and white signals on more than 10% of occasions. Passing the Farnsworth lantern test does not ensure that DCV pilots can distinguish PAPI signal colors. The criterion for passing the lantern test should be made more stringent. In addition the design of PAPI signals can be improved by a better choice of the white color and by providing a redundant non-color cue. abnormal color vision, aviation color vision standard, aviation signal lights, color vision tests, PAPI.


Synthesis of Chiral 1,2-Oxazinanes and Isoxazolidines via Nitroso Aldol Reaction of Distal Dialdehydes.
The first catalytic enantioselective nitroso aldol reaction of distal dialdehydes is reported. The reaction is catalyzed by simple l-proline at room temperature and subsequent reduction delivered biologically potent and synthetically versatile N-O bond containing five- and six-membered heterocycles, 1,2-oxazinanes, and isoxazolidines in high yields and excellent enantioselectivities (up to >99% ee). The method was further exploited to prepare chiral 3-hydroxypiperidines and -pyrrolidines that are otherwise difficult to access.


EVOLUTIONARY IMPLICATIONS OF DNA DIVERGENCE IN THE DROSOPHILA OBSCURA GROUP.
Using DNA-DNA hybridization, we have determined the degree of single-copy DNA (scDNA) divergence among eight species of the Drosophila obscura group. These include Old World and New World species as well as members of two subgroups. Contrary to classical systematics, members of the affinis subgroup are more closely related to American members of the obscura subgroup than are Old World species. The Old World species are not a monophyletic group. The degree of scDNA divergence among species is not necessarily correlated with morphology, chromosomal divergence, or ability to form hybrids. A unique pattern of hybrid formation was found: species separated by a ΔTm of 6.5°C can form hybrids whereas species separated by a ΔTm of 2.5°C cannot. As with other groups of Drosophila, the obscura group has discrete parts of the genome evolving at very different rates. The slow evolving fraction of the nuclear genome is evolving at about the same rate as mitochondrial DNA. The additional scDNA divergence accompanying the step from partial reproductive isolation (between North American pseudoobscura and the isolated Bogotà population) to full isolation is very small. The resolution of the technique was challenged by five closely related taxa with a maximum ΔTm of 2.5°C separating them; the taxa were unambiguously resolved and the "correct" phylogeny recovered. Finally, there is some indication that scDNA in the obscura group may be evolving considerably slower than in the melanogaster subgroup.


Fluoroform-Derived CuCF3 for Trifluoromethylation of Terminal and TMS-Protected Alkynes.
An efficient trifluoromethylation reaction of alkynes using a fluoroform-derived CuCF3 reagent is described. The CF3 source is the inexpensive industrial waste fluoroform (CF3H). The air-stable CuCF3 reagent can be prepared in large quantities and is convenient to use. Synthetically useful trifluoromethylated alkynes containing a wide range of functional groups were successfully synthesized under mild conditions. Both terminal and TMS-protected alkynes gave the products in one step. The beneficial effect of a diamine ligand tetramethylethylenediamine (TMEDA) with the fluoroform-derived CuCF3 reagent was also demonstrated.


Personalized Image Enhancement Using Neural Spline Color Transforms.
In this work we present SpliNet, a novel CNNbased method that estimates a global color transform for the enhancement of raw images. The method is designed to improve the perceived quality of the images by reproducing the ability of an expert in the field of photo editing. The transformation applied to the input image is found by a convolutional neural network specifically trained for this purpose. More precisely, the network takes as input a raw image and produces as output one set of control points for each of the three color channels. Then, the control points are interpolated with natural cubic splines and the resulting functions are globally applied to the values of the input pixels to produce the output image. Experimental results compare favorably against recent methods in the state of the art on the MIT-Adobe FiveK dataset. Furthermore, we also propose an extension of the SpliNet in which a single neural network is used to model the style of multiple reference retouchers by embedding them into a user space. The style of new users can be reproduced without retraining the network, after a quick modeling stage in which they are positioned in the user space on the basis of their preferences on a very small set of retouched images.


Vascular-type disruptive defects in fetuses with homozygous alpha-thalassemia: report of two cases and review of the literature.
The thalassemias are an inherited group of heterogeneous anemias in which one or more of the globin chains in the hemoglobin tetramer are absent. Fetuses with homozygous alpha-thalassemia, which is particularly prevalent in people of Southeast Asian extraction, experience deficient alpha-globin chain synthesis and cannot produce hemoglobin F (the primary fetal hemoglobin after 8 weeks' gestation). Instead, they produce an anomalous hemoglobin, hemoglobin Bart's, with an unusually high affinity for oxygen, leading to profound anemia and tissue hypoxia. Here we report on two fetuses with homozygous alpha-thalassemia who displayed structural defects of a vascular disruptive type. Both fetuses demonstrated limb anomalies, including terminal transverse limb deficiencies, and one fetus was found to have a brain malformation consisting of a neuronal migrational defect. The limb anomalies and suspected brain malformation were detected on prenatal ultrasound prior to confirmation of the diagnosis of alpha-thalassemia in one case; in the other case prenatal records were not available. While microcephaly, hydrocephalus, and retarded brain growth have been rarely reported in association with homozygous alpha-thalassemia, this is the first report of a true brain malformation in an affected fetus. Limb anomalies, on the other hand, appear to be more frequent. Recently, aggressive in utero and postnatal therapies for homozygous alpha-thalassemia have been attempted with some success. Our cases and those from the medical literature suggest that couples need to be counseled about the risks of congenital anomalies of a vascular disruptive type in affected fetuses. Furthermore, data from the literature suggests that in utero therapy may not significantly decrease these risks as such anomalies may be present prior to the institution of therapy. In addition, in hydropic infants with vascular disruptive defects, especially in those of Southeast Asian origin, homozygous alpha-thalassemia should be suspected as a likely etiology.


Identification and Characterization of Botrytis fragariae Isolates on Strawberry in the United States.
Gray mold is a devastating disease on strawberry, and may be caused by several species of Botrytis. The goal of this study was to better understand and characterize the species of Botrytis with reduced sensitivity to the fungicide Polyoxin D, particularly Botrytis fragariae. In total, 78 Botrytis isolates of unknown species that were sensitive (28 isolates; S), moderately sensitive (22 isolates; MS), or reduced sensitive (28 isolates; RS) to Polyoxin-D were collected from commercial strawberry fields of five states in the United States, identified to the species level, and characterized. The majority (75%) of S isolates were Botrytis cinerea and the majority (79%) of RS isolates were the recently described species B. fragariae, indicating an innate ability of B. fragariae to tolerate Polyoxin-D. B. fragariae produced fluffy, white mycelium and was less likely to sporulate on potato dextrose agar than B. cinerea. Isolates from a commercial field recovered from blossoms in early spring were all B. fragariae, those from leaves of the same plants in late spring were a mixture of B. fragariae and B. cinerea, and those from fruit in early summer were all B. cinerea, indicating that B. fragariae may preferentially colonize blossom tissue. A polymerase chain reaction-based assay was developed based on NEP2 sequence variability to distinguish B. fragariae from other Botrytis spp. that have been reported on strawberry, including B. cinerea, B. mali, B. caroliniana, and B. ricini. None of the isolates collected from Canada, California, or North Carolina nurseries were B. fragariae, indicating that the newly described species may not exist or not be widely distributed in planting stock.


67Gallium citrate scintigraphy to assess metastatic spread in a dog with an oral melanoma.
Gallium scintigraphy was used to evaluate therapeutic response in a 10-year-old, male, Dutch sheepdog, suffering from an oral melanoma. Treatment was performed with a combination of carboplatin and hypofractionated radiation. Nineteen weeks after radiation therapy, the left submandibular lymph node was surgically removed because of metastatic disease. Thirty weeks after radiation therapy, 67Gallium scintigraphy was performed to assess for residual disease and metastasis. Increased uptake in the right submandibular lymph node area was noted and identified as a melanoma metastasis on cytology. Surgical excision was performed. Twenty-one weeks later, the dog was euthanased because of advanced pulmonary metastases. This report of a case of oral melanoma illustrates the advantages of 67Gallium scintigraphy in monitoring for the presence of metastatic disease and effectiveness of therapy.


A Randomized Trial of Respiratory Status during Airway Stenting under General Anesthesia Spontaneous Respiration vs. Controlled Ventilation with Muscle Relaxants.
Airway stenting is a procedure in which a stent is inserted into a stenotic site in the airway. However, the optimal ventilation for airway stenting remains controversial. We have planned a randomized, unblinded controlled study to compare intraoperative respiratory status by dividing patients, who underwent airway stenting, into spontaneous respiration (SP) and controlled ventilation with muscle relaxants (MR) groups. This study started in April 2016. The subjects, patients aged ≥20 years with airway stenosis caused by malignant neoplasms for which airway stenting was scheduled, are randomly allocated to SP and MR groups. Anesthesia management is performed in accordance with the anesthetic methods established in each group to compare parameters of the intraoperative respiratory status. The primary endpoint is the incidence of intraoperative oxygen desaturation events (SpO2 <95). Secondary endpoints are the mean intraoperative P/F ratio, pH, PaCO2, adverse events, and proportion of protocol treatment achievement. Currently, there is no evidence of anesthetic methods affecting airway stenting. Some studies have claimed that muscle relaxants worsen airway stenosis, while others have reported stable anesthetic management of controlled ventilation with muscle relaxants in airway stenting. This study may aid in clarifying anesthetic methods for airway stenting.


Estrogen induces glomerulosclerosis in analbuminemic rats.
Progression of chronic renal disease is usually more rapid in males, both in humans and in experimental animals. Estrogen-replacement studies indicate that this may be related to the beneficial effects of estrogen on the lipoprotein profile. However, in hyperlipidemic analbuminemic rats (NAR), females are more prone to develop renal injury than males, and ovariectomy tends to decrease triglyceride levels and prevent renal disease. Therefore, we studied the effects of estrogen administration on lipoproteins, and the induction of renal injury in uninephrectomized female and male NAR. Ovariectomized and orchidectomized uninephrectomized NAR were treated with estradiol implants for 24 weeks. In an additional group of ovariectomized rats, the implant was removed after 12 weeks. Both in ovariectomized and orchidectomized NAR, estradiol caused severe hypercholesterolemia (9 to 12 mmol/liter) and hypertriglyceridemia (6 to 8 mmol/liter) after six weeks. Subsequently, these rats developed severe proteinuria, reaching 209 +/- 25 and 95 +/- 43 mg/day, respectively, after 24 weeks. At this point there was severe glomerular sclerosis, with a respective score of 107 +/- 21 and 61 +/- 33. In terminal blood samples the most pronounced increase in lipid levels were observed in very low density lipoproteins (VLDL), intermediate density lipoproteins (IDL) and low density lipoproteins (LDL). In contrast, ovariectomized NAR and orchidectomized NAR without exogenous estrogen had much lower lipid levels (cholesterol 5 to 7 mmol/liter and triglycerides 1 to 2 mmol/liter) after six weeks. These rats, or ovariectomized NAR where the estrogen treatment had been withdrawn, had practically no proteinuria (4 +/- 1, 19 +/- 11, and 13 +/- 4 mg/day, respectively) or renal damage (glomerulosclerosis score 1 +/- 0.4, 5 +/- 3 and 3 +/- 1, respectively) after 24 weeks. Thus, in hypertriglyceridemic analbuminemic rats, estrogen-treatment causes further increases in both triglycerides and cholesterol. Most probably these changes contribute to the development of renal injury by estrogen in this model. This effect of estrogen, which has also been observed in the Zucker rat, is unique for the hypertriglyceridemic state and deserves further study.


Unilateral onycholysis in a patient taking erlotinib (Tarceva).
Targeting epidermal growth factor receptor (EGFR)-mediated signalling pathways has become routine practice in the treatment of lung cancer. Erlotinib is an oral EGFR tyrosine kinase inhibitor, licensed for maintenance monotherapy treatment in patients with locally advanced or metastatic non-small cell lung cancer after first-line chemotherapy. The authors present the case of a 51-year-old patient who had an excellent response to erlotinib, but developed unilateral onycholysis as an unusual side effect. The authors discuss erlotinib-induced skin and nail changes and have provided a brief literature review on the available evidence for their management.


Strong correlation between the antifungal effect of amphotericin B and its inhibitory action on germ-tube formation in a Candida albicans URA⁺ strain.
The hypothetical capacity of amphotericin B to suppress the formation of germ-tubes, which is the first step of yeast-to-hypha conversion in Candida albicans, has been investigated in the wild-type strain CEY.1 (CAI.4-URA⁺). Exponential cells exposed to concentrations of amphotericin B below or around the MIC90, exhibited a weak reduction in the percentage of human serum-induced germ-tube formation at 37ºC compared with a non-exposed control. However, the dimorphic transition was drastically suppressed after addition of potentially lethal doses of amphotericin B, which also caused severe cell killing. In contrast, an identical experimental approach carried out with the fungistatic compound 5-fluorocytosine had no significant effect on the level of the germ-tube formation. Together, these results strongly point to a close correlation between the fungicidal action of amphotericin B and its ability to impair morphogenetic conversion in C. albicans.


[Frequency of antiphospholipid antibodies and factor V (G1691A), prothrombin (G20210A) gene polimorphism among women with pregnancy complications].
Thrombophilia these days is a subject of many medical research including obstetric and gynecology where causing feto-maternal complications. In women predispose to venous thromboembolism in high risk situation like pregnancy, puerperium, operation, prolonged bed rest or hormonal treatment. For fetal complication account miscarriages, intrauterine deaths, IUGR, and for maternal account premature placental separation and severe preeclampsia. Diagnostic panel include inherited factors like factor V Leiden and prothrombin mutation, activated protein C resistance and acquired like anticardiolipin antibodies, antibodies against beta2-glicoprotein 1 and lupus anticoagulant. The aim of this paper is estimation ofthrombophilia as a causative factor of pregnancy complications and attempt to establish screening criteria for thrombophilia. Material involved 36 women with pregnancy complications divided into three groups correlating with trimester when pregnancy loss occured. All patients had genetic, hormonal and anatomic tests done on purpose to exclude other possible causes of miscarriages. All women had done both types of tests for inherited and acquired thrombophilia. For factor V and II gene polimorphism we used PCR and RLFP method. Acquired protein C resistance and lupus anticoagulant was tested using chronometric method with the use of time measurement of optical density accompanying coagulation. Anticardiolipin antibodies and antibodies against beta2-glicoprotien 1 were measured in ELISA tests. Our current results present the frequency of at least one thrombophilic factor in 16.6% patients. The highest frequency rate was observed among women with pregnancy loss between 7th and 12th gestational week--19.04%. In this group we also noticed the highest number of miscarriages. In remaining two groups, with pregnancy loss between 12th and 22nd and after 22nd gestational week, one case of thrombophilia occurred in each group.


Seminal cell-free DNA levels measured by PicoGreen fluorochrome are associated with sperm fertility criteria.
Previous investigations suggested that elevated cell-free DNA (cfDNA) can indicate non-healthy states. However, the potential association between cfDNA seminal plasma levels and fertility sperm parameters has not yet been determined. Therefore, the present study evaluated the association between seminal cfDNA levels and sperm fertility criteria to determine the use of seminal cfDNA quantification. An in vivo protocol quantified cfDNA levels of semen samples obtained from 163 male patients using fluorescent PicoGreen dye staining. To confirm if semen cfDNA quantification is realistic, an in vitro complementary test was performed using three or four semen samples. The fresh sperm samples were exposed to paraquat that generates high levels of superoxide anion causing oxidative stress and cell mortality. The results showed significant association between dsDNA levels and several sperm fertility parameters, such as low viability and alterations of motility and morphology. The in vitro analysis confirmed the association between dsDNA levels and sperm viability. Together, these results suggest that dsDNA levels could be an important biomarker to test sperm fertility.


Mitochondrial DNA and primary mitochondrial dysfunction in Parkinson's disease.
In 1979, it was observed that parkinsonism could be induced by a toxin inhibiting mitochondrial respiratory complex I. This initiated the long-standing hypothesis that mitochondrial dysfunction may play a key role in the pathogenesis of Parkinson's disease (PD). This hypothesis evolved, with accumulating evidence pointing to complex I dysfunction, which could be caused by environmental or genetic factors. Attention was focused on the mitochondrial DNA, considering the occurrence of mutations, polymorphic haplogroup-specific variants, and defective mitochondrial DNA maintenance with the accumulation of multiple deletions and a reduction of copy number. Genetically determined diseases of mitochondrial DNA maintenance frequently manifest with parkinsonism, but the age-related accumulation of somatic mitochondrial DNA errors also represents a major driving mechanism for PD. Recently, the discovery of the genetic cause of rare inherited forms of PD highlighted an extremely complex homeostatic control over mitochondria, involving their dynamic fission/fusion cycle, the balancing of mitobiogenesis and mitophagy, and consequently the quality control surveillance that corrects faulty mitochondrial DNA maintenance. Many genes came into play, including the PINK1/parkin axis, but also OPA1, as pieces of the same puzzle, together with mitochondrial DNA damage, complex I deficiency and increased oxidative stress. The search for answers will drive future research to reach the understanding necessary to provide therapeutic options directed not only at limiting the clinical evolution of symptoms but also finally addressing the pathogenic mechanisms of neurodegeneration in PD. © 2017 International Parkinson and Movement Disorder Society.