Far-infrared ray for treating chronic lower extremity lymphedema with dermatolymphangioadenitis: a postoperative complication of gynecological tumor resection.
Lower extremity lymphedema is regarded as a relatively common postoperative complication and is often accompanied with dermatolymphangioadenitis (DLA). This study combines clinical assessment and laboratory investigation to explore therapeutic effects of far-infrared radiation (FIR) therapy for chronic lower extremity lymphedema accompanied with DLA, occurring after gynecological tumor resection. Patients who met inclusion and exclusion criteria would be enrolled. They received regular sessions using the FIR therapy machine over the 4-week treatment course. Clinical and laboratory outcome measures were carried out before and after treatment. Clinical outcome measures included DLA seizure frequency (episodes/year), patients' subjective feedback for lymphedema-related symptoms and quality of life (QOL). Laboratory outcome measures included bacterial cultures and concentrations of inflammatory cytokines: IL-1β, IL-2, IL-4, IL-10, IL-12, IL-18, TNF-α, TNF-β, caspase-1 and INF-γ, detected in serum and local lymphedema tissue fluid samples using protein microarray and ELISA. Between 2012 and 2016, a total of 120 female patients were screened for study enrollment. Sixty-four recruited patients underwent clinical evaluation both before FIR radiation therapy and 1 year after a single course of FIR radiation therapy. Eleven patients (17.2%), randomly chosen from the study group, underwent additional laboratory analysis of blood and local lymphedema tissue fluid samples. The frequency of DLA decreased following treatment (p < 0.001). Fifty patients (78%) did not experience a single episode of DLA recurrence in the year subsequent to treatment. The efficiency rate calculated using DLA frequencies was greater than 50% for 63 (98%) patients following treatment. Patients reported a subjective decrease in lymphedema-related symptoms (p < 0.05). Patients' QOL scores were higher after treatment (p < 0.001). Laboratory analysis showed an elevation in serum concentration of IL-1β after FIR therapy (p < 0.05) and reduced local tissue fluid concentrations of inflammatory cytokines IL-2, IL-10 and IL-18 (p < 0.05). Bacterial culture results before and after treatment were both negative. FIR radiation therapy provides an effective treatment modality for patients with chronic lymphedema accompanied with DLA that develops secondarily to treatment of gynecological malignancies, whose therapeutic effects may be due to reduced immune dysfunction within local lymphedema tissues.


Separation of americium from curium by oxidation and ion exchange.
Nuclear energy has the potential to be a clean alternative to fossil fuels, but in order for it to play a major role in the US, many questions about the back end of the fuel cycle must be addressed. One of these questions is the difficult separation of americium from curium. Here, we report the oxidation of Am in two systems, perchloric acid and nitric acid and the affect of changing the acid has on the oxidation. K(d) values were observed and a direct separation factor was calculated and was seen to be as high as 20 for four metal(IV) pillared phosphate phosphonate inorganic organic hybrid ion exchange materials. These ion exchangers are characterized by very low selectivity for cations with low charge but extremely high uptake of ions of high charge.


Modeling of biomolecular machines in non-equilibrium steady states.
Numerical computations have become a pillar of all modern quantitative sciences. Any computation involves modeling-even if often this step is not made explicit-and any model has to neglect details while still being physically accurate. Equilibrium statistical mechanics guides both the development of models and numerical methods for dynamics obeying detailed balance. For systems driven away from thermal equilibrium, such a universal theoretical framework is missing. For a restricted class of driven systems governed by Markov dynamics and local detailed balance, stochastic thermodynamics has evolved to fill this gap and to provide fundamental constraints and guiding principles. The next step is to advance stochastic thermodynamics from simple model systems to complex systems with tens of thousands or even millions of degrees of freedom. Biomolecules operating in the presence of chemical gradients and mechanical forces are a prime example for this challenge. In this Perspective, we give an introduction to isothermal stochastic thermodynamics geared toward the systematic multiscale modeling of the conformational dynamics of biomolecular and synthetic machines, and we outline some of the open challenges.


TelecomNet: Tag-Based Weakly-Supervised Modally Cooperative Hashing Network for Image Retrieval.
We are concerned with using user-tagged images to learn proper hashing functions for image retrieval. The benefits are two-fold: (1) we could obtain abundant training data for deep hashing models; (2) tagging data possesses richer semantic information which could help better characterize similarity relationships between images. However, tagging data suffers from noises, vagueness and incompleteness. Different from previous unsupervised or supervised hashing learning, we propose a novel weakly-supervised deep hashing framework which consists of two stages: weakly-supervised pre-training and supervised fine-tuning. The second stage is as usual. In the first stage, we propose two formulations Tag-basEd weakLy-supErvised Modally COoperative hashing Network (TelecomNet) and Generalized TelecomNet (GTelecomNet). Rather than performing supervision on tags, TelecomNet first learns an observed semantic embedding vector for each image from attached tags and then uses it to guide hashing learning. GTelecomNet introduces a novel semantic network to exploit more precise semantic information. By carefully designing the optimization problem, they can well leverage tagging information and image content for hashing learning. The framework is general and does not depend on specific deep hashing methods. Empirical results on real world datasets show that they significantly increase the performance of state-of-the-art deep hashing methods.


[Comparison of the effects of midazolam and thiopental on the neuromuscular response of vecuronium, atracurium, and pancuronium].
To assess the possible interactions of midazolam and thiopental with the muscular relaxants vecuronium (0.08 mg/kg), atracurium (0.5 mg/kg), and pancuronium (0.1 mg/kg), a comparative analysis was undertaken in two groups of 18 and 32 patients treated respectively with midazolam (0.3 mg/kg) and thiopental (5 mg/kg). The beginning of the effect, maximal blockade, duration of the clinical response, and the spontaneous recovery index were measured on electromyographic recordings of action potentials evoked by train of four supramaximal stimuli delivered every 20 sec on the ulnar nerve. Conditions for intubation were assessed 2 minutes after administration of muscular relaxant. There were no significant differences in neuromuscular parameters in either of the two groups of patients treated with midazolam or with thiopental independently of the relaxant drug administered.


Amplified Striatal Responses to Near-Miss Outcomes in Pathological Gamblers.
Near-misses in gambling games are losing events that come close to a win. Near-misses were previously shown to recruit reward-related brain regions including the ventral striatum, and to invigorate gambling behavior, supposedly by fostering an illusion of control. Given that pathological gamblers are particularly vulnerable to such cognitive illusions, their persistent gambling behavior might result from an amplified striatal sensitivity to near-misses. In addition, animal studies have shown that behavioral responses to near-miss-like events are sensitive to dopamine, but this dopaminergic influence has not been tested in humans. To investigate these hypotheses, we recruited 22 pathological gamblers and 22 healthy controls who played a slot machine task delivering wins, near-misses and full-misses, inside an fMRI scanner. Each participant played the task twice, once under placebo and once under a dopamine D2 receptor antagonist (sulpiride 400 mg), in a double-blind, counter-balanced design. Participants were asked about their motivation to continue gambling throughout the task. Across all participants, near-misses elicited higher motivation to continue gambling and increased striatal responses compared with full-misses. Crucially, pathological gamblers showed amplified striatal responses to near-misses compared with controls. These group differences were not observed following win outcomes. In contrast to our hypothesis, sulpiride did not induce any reliable modulation of brain responses to near-misses. Together, our results demonstrate that pathological gamblers have amplified brain responses to near-misses, which likely contribute to their persistent gambling behavior. However, there is no evidence that these responses are influenced by dopamine. These results have implications for treatment and gambling regulation.


Melatonin elicits protein kinase C-mediated calcium response in immortalized GT1-7 GnRH neurons.
Melatonin is suggested to have effects on hypothalamic-pituitary-gonadal (HPG) axis. The pulsatile pattern of GnRH release, which results in the intermittent release of gonadotropic hormones from the pituitary, has a critical importance for reproductive function but the factors responsible from this release pattern are not known. Calcium is a second messenger involved in hormone release. Therefore, investigation of the effects of melatonin on intracellular free calcium levels ([Ca(2+)](i)) would provide critical information on hormone release in immortalized GnRH neurons. The pattern of melatonin-induced intracellular calcium signaling was investigated by fluorescence calcium imaging using the immortalized GnRH-secreting GT1-7 hypothalamic neurons. Melatonin caused a significant increase in [Ca(2+)](i,) which was greatly blocked by luzindole, a melatonin antagonist, or attenuated by pre-treatment with protein kinase C inhibitor. This study suggests that melatonin seems to have a direct effect on GnRH neurons.


Rare-Earth-Free Magnets: Enhancing Magnetic Anisotropy and Spin Exchange Toward High-TC Hf2MIr5B2 (M = Mn, Fe).
Designing new rare-earth-free (REF) permanent magnetic materials (PMM) to replace the high performing but critically restrained rare-earth-based PMM remains a great challenge to the scientific community. Here, we report on the rational design of new REF PMM, Hf2MIr5B2 (M = Fe, Mn) via a theory-experiment combined approach. Density functional theory (DFT) predicted strong interchain M-M spin-exchange coupling and large magnetocrystalline anisotropy energies (EMAE) for the new compounds, suggesting potential intrinsic PMM properties. Subsequent experimental bulk syntheses and magnetic characterizations established the highest ordering temperature (TC ∼ 900 K) for Hf2FeIr5B2 and the highest intrinsic coercivity (HC) value for Hf2MnIr5B2 (HC = 62.1 kA/m) reported to date for Ti3Co5B2-type compounds. Importantly, at room temperature both phases show significant coercivities due to intrinsic factors only, hinting at their huge potential to create REF PMM by improving extrinsic factors such as controlling the microstructure and the domain orientation.


Expression and functional activity of four myocardin isoforms.
Myocardin (MYOCD) is an essential component of a molecular switch for the expression of contractile genes in smooth muscle and cardiac muscle cells. The Myocd gene comprises at least fifteen exons, including two alternately spliced exons designated 2a and 10a. We investigated tissue-specific Myocd expression in mouse, rat and human tissues to determine the conservation in expression of each Myocd splice variant and to ascertain whether any functional differences exist among MYOCD isoforms. Conventional and quantitative RT-PCR revealed the dominant expression of Myocd exon 2a (Myocd_v3) in smooth muscle cell (SMC)-rich tissues (aorta and bladder) with little expression in heart across all species studied. Each species of heart showed primarily a longer version of Myocd (Myocd_v1) without exon 2a. While exclusion of exon 2a was common in all cardiac muscle samples, exon skipping of Myocd exon 10a was a rare event in both cardiac muscle and SMC tissues. In general, all four MYOCD isoforms showed comparable stimulation of SMC promoters. On the other hand, Myocd_v1 and Myocd_v2 were more active than Myocd_v3 and Myocd_v4 in stimulating cardiac muscle promoters and Myocd_v1's activity was augmented in the presence of the cardiac transcription factor, MEF2C. Importantly, whereas all four MYOCD isoforms similarly induced expression of endogenous SMC genes in a prostate tumor cell line (LNCaP), none could induce the endogenous expression of specific cardiac markers. These results are the first to show relative expression and activities of the major myocardin isoforms across disparate species. We propose a new myocardin nomenclature reflecting the dominant splice variants expressed in cardiac muscle (Myocd_v1 and v2) versus SMC-rich tissues (Myocd_v3 and v4).


Burden of infection on growth failure.
The high prevalence of infections among children living in poor areas of developing countries impairs linear growth in these populations. Acute, invasive infections, which provoke a systemic response (e.g., dysentery and pneumonia), and chronic infections, which affect the host over a sustained period (e.g., gut helminth infections), have a substantial effect on linear growth. Such infections can diminish linear growth by affecting nutritional status. This occurs because infections may decrease food intake, impair nutrient absorption, cause direct nutrient losses, increase metabolic requirements or catabolic losses of nutrients and, possibly, impair transport of nutrients to target tissues. In addition, induction of the acute phase response and production of proinflammatory cytokines may directly affect the process of bone remodeling that is required for long bone growth. Infection of cells directly involved in bone remodeling (osteoclasts or osteoblasts) by specific viruses may also directly affect linear growth. Many interventions are possible to diminish the effect of infection on growth. Prevention of disease through sanitation, vector control, promotion of breast-feeding and vaccination is crucial. Appropriate treatment of infections (e.g., antibiotics for pneumonia) as well as supportive nutritional therapy (again including breast-feeding) during and after recovery, is also important. Targeted therapeutic interventions to decrease the prevalence of gut helminth infections may also be appropriate in areas in which such infections are widespread. Such interventions are of public health benefit not only because they reduce the incidence or severity of infections, but also because they decrease the long-term detrimental effect of malnutrition on populations.


Abnormalities in CD57+ cytotoxic T cells and Vδ1+ γδT cells in subclinical celiac disease in childhood are affected by cytomegalovirus. The Generation R Study.
Celiac disease (CD) is a digestive and autoimmune disorder driven by an immune response to modified gluten peptides. Affected intestines show infiltrates of various T-cell and NK-cell subsets. It is currently unclear if individuals with subclinical CD have systemic abnormalities in immune cells. We here studied whether subclinical CD is associated with changes in blood CD57-expressing and Vδ1-expressing lymphocytes in children, and whether cytomegalovirus (CMV) infection modifies this association. Included were 1068 children from the Generation R Study. Serum Immunoglobulin G (IgG) levels against CMV were measured by ELISA; Tissue transglutaminase type 2 antibody (TG2A) levels with fluorescence enzyme immunoassay (FEIA). Duodenal biopsies, additional Human Leukocyte Antigen (HLA) DQ 2.2, 2.5 and 8 and endomysial antibody (EMA) typing were performed in TG2A positive children. Subclinical CD cases (n=12) had 1.8 fold (95% CI 1.06; 3.1) fewer Vδ1+ T cells which was predominantly observed in CMV seronegative children (p-interaction 0.02), and 2.7 fold (95% CI 1.25; 5.99) more CD57+ T cells than HLA DQ2/-DQ8 positive controls (n=339). Hence, children with subclinical CD have alterations in specific blood T cell subsets that are linked to viral pathology. The observed interaction effect between subclinical CD and CMV may contribute to the understanding of disease pathogenesis.


Knee function after anterior cruciate ligament injury in elite collegiate athletes.
Anterior cruciate ligament injuries are common in athletes, but there are few studies of long-term outcomes. Long-term knee function of anterior cruciate ligament-injured athletes is inferior to that of their uninjured teammates. Retrospective cohort study. Thirty-three Division I-A athletes who had sustained an anterior cruciate ligament injury during their college career completed a series of questionnaires that assessed knee function and quality of life 2 to 14 years after injury. Their responses were compared with those of a matched cohort of their uninjured teammates. There were no differences in the mean Tegner scores, modified Lysholm scores, or in the scores of the SF-36 between groups. Sixteen anterior cruciate ligament-injured athletes scored A or B in the subjective portion of the International Knee Documentation Committee score and 17 scored C or D, whereas 24 control subjects scored A or B and 9 scored C or D, a statistically significant difference between groups. Five injured and 14 control athletes had participated at a professional or national team level after college. Quality of life of elite collegiate athletes who sustained an anterior cruciate ligament injury was not significantly different from that of their uninjured teammates, but knee function differed between groups.


Influenza vaccination coverage among persons with asthma--United States, 2005-06 influenza season.
During 2006, approximately 6.8 million (9.3%) U.S. children and 16.1 million (7.3%) U.S. adults were reported to have asthma. Since 1964, the Advisory Committee on Immunization Practices (ACIP) has recommended influenza vaccination of all persons with asthma because of the higher risk for medical complications from influenza for those persons. Influenza vaccination coverage of persons with asthma varies by age group and remains below Healthy People 2010 targets of 60% coverage of persons aged 18--64 years with high-risk conditions (14-29 c) and 90% of all persons aged > or =65 years (14-29 a). Influenza vaccination rates of children and older adults with asthma have not been well studied. Using 2006 National Health Interview Survey (NHIS) data, this report provides the first examination of influenza vaccination rates and related factors across a national sample of persons with asthma aged > or =2 years. The results indicated that 36.2% received influenza vaccination during the 2005--06 influenza season. Vaccination rates remained below target levels among all subgroups examined, including those reporting the greatest number of health-care visits in the past 12 months. The results of this study indicate that influenza vaccination coverage of all persons with asthma can be improved by increasing access to health care and using opportunities for vaccination during health-care visits.


Stable 5,6-epoxyeicosatrienoic acid analog relaxes coronary arteries through potassium channel activation.
5,6-epoxyeicosatrienoic acid (5,6-EET) is a cytochrome P450 epoxygenase metabolite of arachidonic acid that causes vasorelaxation. However, investigations of its role in biological systems have been limited by its chemical instability. We developed a stable agonist of 5,6-EET, 5-(pentadeca-3(Z),6(Z),9(Z)-trienyloxy)pentanoic acid (PTPA), in which the 5,6-epoxide was replaced with a 5-ether. PTPA obviates chemical and enzymatic hydrolysis. In bovine coronary artery rings precontracted with U46619, PTPA (1 nmol/L to 10 micromol/L) induced concentration-dependent relaxations, with maximal relaxation of 86+/-5% and EC50 of 1 micromol/L. The relaxations were inhibited by the cyclooxygenase inhibitor indomethacin (10 micromol/L; max relaxation 43+/-9%); the ATP-sensitive K+ channel inhibitor glybenclamide (10 micromol/L; max relaxation 49+/-6%); and the large conductance calcium-activated K+ channel inhibitor iberiotoxin (100 nmol/L; max relaxation 38+/-6%) and abolished by the combination of iberiotoxin with indomethacin or glybenclamide or increasing extracellular K+ to 20 mmol/L. Whole-cell outward K+ current was increased nearly 6-fold by PTPA (10 micromol/L), which was also blocked by iberiotoxin. Additionally, we synthesized 5-(pentadeca-6(Z),9(Z)-dienyloxy)pentanoic acid and 5-(pentadeca-3(Z),9(Z)-dienyloxy)pentanoic acid (PDPA), PTPA analogs that lack the 8,9 or 11,12 double bonds of arachidonic acid and therefore are not substrates for cyclooxygenase. The PDPAs caused concentration-dependent relaxations (max relaxations 46+/-13% and 52+/-7%, respectively; EC50 1micromol/L), which were not altered by glybenclamide but blocked by iberiotoxin. These studies suggested that PTPA induces relaxation through 2 mechanisms: (1) cyclooxygenase-dependent metabolism to 5-ether-containing prostaglandins that activate ATP-sensitive K+ channels and (2) activation of smooth muscle large conductance calcium-activated K+ channels. PDPAs only activate large conductance calcium-activated K+ channels.


Apical segmental resection osteotomy with dual axial rotation corrective technique for severe focal kyphosis of the thoracolumbar spine.
in this paper, the authors' goal was to evaluate the feasibility, safety, and efficacy of apical segment resection osteotomy with dual axial rotation correction for severe focal kyphosis by examining outcomes. between May 2004 and December 2006, the authors treated 23 patients with severe focal kyphosis (average Cobb angle 86.9°, range 50°-130°) using apical segmental resection osteotomy with dual axial rotation correction and instrumented anterior column reconstruction and fusion. Radiographic assessment of sagittal plane balance and kyphotic Cobb angle (including a scoliosis Cobb angle in 9 cases) was performed in each patient before and immediately after surgery and at the last follow-up (minimum 2 years). The Frankel grading system for neurological function and Oswestry Disability Index for quality of life were evaluated before surgery and at the last follow-up. The patient satisfaction index was also used for clinical evaluation at the last follow-up. the mean surgical time was 6.7 hours. The average blood loss was 2960 ml. All patients underwent follow-up for 2 or more years after surgery. The fusion rate was 95.65%. The average kyphotic angle improved from 86.9° preoperatively to 25.6° immediately postoperatively, with an average correction rate of 72.17%. At the last follow-up, the average kyphotic angle was 27.4°, making the final correction rate 69.87%. The sagittal plane balance was significantly improved at the last follow-up. Preoperatively, 15 patients had neurological deficits, and the Frankel grade was E in 8 cases, D in 8 cases, C in 6 cases, and B in 1 case. At the last follow-up, 15 cases were Grade E, 5 were Grade D, and 3 were Grade C. The average improvement in the Oswestry Disability Index score was 43.30%. The patient satisfaction index result showed a total satisfaction rate of 91.30%. Complications included 1 case of late neurological deficit due to shifting of an expandable artificial vertebra, 5 cases of nerve root injury, 3 cases of dural tear, and 1 case of transient lower-extremity weakness due to insufficient blood supply to the spinal cord during surgery. apical segmental resection osteotomy with dual axial rotation correction and instrumented fusion is an effective and safe way to treat severe focal kyphosis of the thoracolumbar spine.


Long non-coding RNA HOTAIR promotes carcinogenesis and invasion of gastric adenocarcinoma.
Gastric cancer is one of the major causes of cancer death worldwide; however, the mechanism of carcinogenesis is complex and poorly understood. Long non-coding RNA HOTAIR (HOX transcript antisense RNA) recently emerged as a promoter of metastasis in various cancers including gastric cancer. Here we investigated the impact of HOTAIR on apoptosis, cell proliferation and cell cycle to dissect the carcinogenesis of gastric cancer. We examined the mechanism of invasion and metastasis and analyzed the clinical significance of HOTAIR. Downregulation of HOTAIR was confirmed by two different siRNAs. The expression of HOTAIR was significantly elevated in various gastric cancer cell lines and tissues compared to normal control. si-HOTAIR significantly reduced viability in MKN 28, MKN 74, and KATO III cells but not in AGS cells. si-HOTAIR induced apoptosis in KATO III cells. Lymphovascular invasion and lymph node metastasis were more common in the high level of HOTAIR group. si-HOTAIR significantly decreased invasiveness and migration. si-HOTAIR led to differential expression of epithelial to mesenchymal transition markers. We found that HOTAIR was involved in inhibition of apoptosis and promoted invasiveness, supporting a role for HOTAIR in carcinogenesis and progression of gastric cancer.


Plastid dsRNA transgenes trigger phased small RNA-based gene silencing of nuclear-encoded genes.
Plastid transformation technology has been widely used to express traits of potential commercial importance, though the technology has been limited to traits that function while sequestered in the organelle. Prior research indicates that plastid contents can escape from the organelle, suggesting a possible mechanism for engineering plastid transgenes to function in other cellular locations. To test this hypothesis, we created tobacco (Nicotiana tabacum cv. Petit Havana) plastid transformants that express a fragment of the nuclear-encoded Phytoene desaturase (PDS) gene capable of catalyzing post-transcriptional gene silencing if RNA escapes into the cytoplasm. We found multiple lines of direct evidence that plastid-encoded PDS transgenes affect nuclear PDS gene silencing: knockdown of the nuclear-encoded PDS mRNA and/or its apparent translational inhibition, biogenesis of 21-nucleotide (nt) phased small interfering RNAs (phasiRNAs), and pigment-deficient plants. Furthermore, plastid-expressed dsRNA with no cognate nuclear-encoded pairing partner also produced abundant 21-nt phasiRNAs in the cytoplasm, demonstrating that a nuclear-encoded template is not required for siRNA biogenesis. Our results indicate that RNA escape from plastids to the cytoplasm occurs generally, with functional consequences that include entry into the gene silencing pathway. Furthermore, we uncover a method to produce plastid-encoded traits with functions outside of the organelle and open additional fields of study in plastid development, compartmentalization, and small RNA biogenesis.


Parental influences on adolescent decision making and contraceptive use.
This article reviews the literature in the area of parental and maternal influences on contraceptive decision making, thus determining future directions of research and potential interventions. A review of published literature using PubMed, CINAHL, Infotrak, Science and Technology, and Medline spanning 1980-2007 was used to gather information. Results from 35 research studies and 15 scholarly articles were included. The search terms adolescent decision making, maternal influences on adolescent contraceptive decision making, adolescent contraceptive decision making, paternal influences on adolescent decision making, and contraception were used to gather data. The literature reveals there is an association between parental communication, parenting style, and adolescent sexual activity and contraception use. Maternal communication has been shown to delay sexual intercourse and increase contraceptive use. Maternal communication has rich potential as an intervention to impact positive adolescent sexual decision making and contraception use. Gaps in the research identified were the lack of studies on the influences on parenting style and the father/adolescent influences on sexual initiation and contraceptive decision making.


The longevity gene INDY (I'm Not Dead Yet) in metabolic control: Potential as pharmacological target.
The regulation of metabolic processes by the Indy (I'm Not Dead Yet) (SLC13A5/NaCT) gene was revealed through studies in Drosophila melanogaster and Caenorhabditis elegans. Reducing the expression of Indy in these species extended their life span by a mechanism resembling caloric restriction, without reducing food intake. In D. melanogaster, mutating the Indy gene reduced body fat content, insulin-like proteins and reactive oxygen species production. Subsequent studies indicated that Indy encodes a citrate transporter located on the cell plasma membrane. The transporter is highly expressed in the mammalian liver. We generated a mammalian knock out model deleting the mammalian homolog mIndy (SLC13A5). The knock out animals were protected from HFD induced obesity, fatty liver and insulin resistance. Moreover, we have shown that inducible and liver selective knock down of mIndy protects against the development of fatty liver and insulin resistance and that obese humans with type 2 diabetes and non-alcoholic fatty liver disease have increased levels of mIndy. Therefore, the transporter mINDY (NaCT) has been proposed to be an 'ideal target for the treatment of metabolic disease'. A small molecule inhibitor of the mINDY transporter has been generated, normalizing glucose levels and reducing fatty liver in a model of diet induced obese mice. Taken together, studies from lower organisms, mammals and humans suggest that mINDY (NaCT) is an attractive target for the treatment of metabolic disease.


Chemical defense and antifouling activity of three Mediterranean sponges of the genus Ircinia.
The defense roles and the antifouling activity of the organic extracts and the major metabolites of the sponges Ircinia oros, I. variabilis and I. spinosula were investigated. The antifeedant activity was tested in experimental aquaria on the generalist predator fish Thalassoma pavo as well as in coastal ecosystems rich in fishes. Some of the major metabolites exhibited high levels of antifeedant activity. The antifouling activity was tested in laboratory assays, against representatives of the major groups of fouling organisms (marine bacteria, marine fungi, diatoms, macroalgae and mussels). All extracts showed promising levels of activity. As was expected, no single extract was active in all tests and some fractions that were effective against one organism showed little or no activity against the others. The high but variable level of antifouling activity in combination with the absence of toxicity (tested on the development of oyster and sea urchin larvae) shows the potential of these metabolites to become ingredients in environmentally friendly antifouling preparations.


Reliability and minimal detectable change of the mini-BESTest in adults with spinal cord injury in a rehabilitation setting.
Background: The mini-Balance Evaluation Systems Test (mini-BESTest) is a valid tool for assessing standing balance in people with spinal cord injury (SCI). Its reliability has not yet been investigated with this population. Objective: To assess the test-retest and inter-rater reliability of the mini-BESTest in adults with SCI in a rehabilitation setting. Methods: Twenty-three participants admitted in a rehabilitation center following an SCI (mean age = 52.2 years, SD = 14.5; 13/23 tetraplegia; 14/23 traumatic injury) and able to stand 30 seconds without help were recruited. They were evaluated twice with the mini-BESTest to establish the test-retest reliability (interval of 1 to 2 days). One of the two sessions was video-recorded to establish the inter-rater reliability (3 physiotherapists). Intraclass correlation coefficients (ICC2,1), weighted kappa (Kw) and Kendall's W were used to determine reliability of total score and individual items. Minimal detectable changes (MDC) were computed. Results. The mini-BESTest total scores showed excellent test-retest (ICC = 0.94) and inter-rater (ICC = 0.96) reliability. Reliability of 50% of the individual items was acceptable to excellent (Κw and W = 0.35-1.00). The MDC of the mini-BESTest total score was 4 points. Conclusion: The mini-BESTest is a reliable tool to assess standing balance in adults with an SCI. A minimal change of 4 points on the total scale is needed to be confident that the change is not a measurement error between two sessions or two raters.


Thrombolytic therapy in peripheral arterial occlusive disease: mechanisms of action and drugs available.
Catheter-directed thrombolytic therapy has become an important part of the treatment of patients with acute arterial and graft occlusion. The underlying pharmacologic principle is the activation of plasminogen, bound to fibrin within the thrombus. Guide-wire passage reliably predicts success of catheter-directed thrombolysis. The underlying disease process leading to thrombosis should be accurately identified and promptly corrected to reduce the probability of recurrent occlusion. Streptokinase (SK), urokinase (UK) and recombinant tissue plasminogen activator (rt-PA) are the three agents used to treat peripheral arterial occlusive disease. The evolution from SK to UK and rt-PA and improvements in techniques and delivery systems have led to improved success rates and lower complication rates. Patient selection, basic technical considerations and overall results are discussed here. The currently available thrombolytic agents, as well as those being developed, are reviewed to provide background information for current and future applications.


Laetrile treatment for cancer.
Laetrile is the name for a semi-synthetic compound which is chemically related to amygdalin, a cyanogenic glycoside from the kernels of apricots and various other species of the genus Prunus. Laetrile and amygdalin are promoted under various names for the treatment of cancer although there is no evidence for its efficacy. Due to possible cyanide poisoning, laetrile can be dangerous. To assess the alleged anti-cancer effect and possible adverse effects of laetrile and amygdalin. We searched the following databases: CENTRAL (2014, Issue 9); MEDLINE (1951-2014); EMBASE (1980-2014); AMED; Scirus; CINAHL (all from 1982-2015); CAMbase (from 1998-2015); the MetaRegister; the National Research Register; and our own files. We examined reference lists of included studies and review articles and we contacted experts in the field for knowledge of additional studies. We did not impose any restrictions of timer or language. Randomized controlled trials (RCTs) and quasi-RCTs. We searched eight databases and two registers for studies testing laetrile or amygdalin for the treatment of cancer. Two review authors screened and assessed articles for inclusion criteria. We located over 200 references, 63 were evaluated in the original review, 6 in the 2011 and none in this update. However, we did not identify any studies that met our inclusion criteria. The claims that laetrile or amygdalin have beneficial effects for cancer patients are not currently supported by sound clinical data. There is a considerable risk of serious adverse effects from cyanide poisoning after laetrile or amygdalin, especially after oral ingestion. The risk-benefit balance of laetrile or amygdalin as a treatment for cancer is therefore unambiguously negative.


Identification of an NTF2-related factor that binds Ran-GTP and regulates nuclear protein export.
Active transport of macromolecules between the nucleus and cytoplasm requires signals for import and export and their recognition by shuttling receptors. Each class of macromolecule is thought to have a distinct receptor that mediates the transport reaction. Assembly and disassembly reactions of receptor-substrate complexes are coordinated by Ran, a GTP-binding protein whose nucleotide state is regulated catalytically by effector proteins. Ran function is modulated in a noncatalytic fashion by NTF2, a protein that mediates nuclear import of Ran-GDP. Here we characterize a novel component of the Ran system that is 26% identical to NTF2, which based on its function we refer to as NTF2-related export protein 1 (NXT1). In contrast to NTF2, NXT1 preferentially binds Ran-GTP, and it colocalizes with the nuclear pore complex (NPC) in mammalian cells. These properties, together with the fact that NXT1 shuttles between the nucleus and the cytoplasm, suggest an active role in nuclear transport. Indeed, NXT1 stimulates nuclear protein export of the NES-containing protein PKI in vitro. The export function of NXT1 is blocked by the addition of leptomycin B, a compound that selectively inhibits the NES receptor Crm1. Thus, NXT1 regulates the Crm1-dependent export pathway through its direct interaction with Ran-GTP.


Identifying potential sources of variability between vegetation carbon storage estimates for urban areas.
Although urbanisation is a major cause of land-use change worldwide, towns and cities remain relatively understudied ecosystems. Research into urban ecosystem service provision is still an emerging field, yet evidence is accumulating rapidly to suggest that the biological carbon stores in cities are more substantial than previously assumed. However, as more vegetation carbon densities are derived, substantial variability between these estimates is becoming apparent. Here, we review procedural differences evident in the literature, which may be drivers of variation in carbon storage assessments. Additionally, we quantify the impact that some of these different approaches may have when extrapolating carbon figures derived from surveys up to a city-wide scale. To understand how/why carbon stocks vary within and between cities, researchers need to use more uniform methods to estimate stores and relate this quantitatively to standardised 'urbanisation' metrics, in order to facilitate comparisons.


Successful delivery in a female with thrombotic thrombocytopenic purpura.
A 28-year-old female, who suffered from thrombotic thrombocytopenic purpura (TTP) in the 14th week of her first pregnancy, recovered after a plasma exchange followed by an induced abortion. From six months after the abortion, she no longer required plasma infusions every 3-4 weeks to prevent a relapse of TTP manifested as thrombocytopenia, and complete remission continued until her next pregnancy. In her second pregnancy, she had an immediate relapse of TTP and responded to plasma infusion until the 24th week. However, the TTP gradually became resistant to plasma infusion, and developed into toxemia with edema, hypertension and proteinuria in the 27th week. Although the TTP was alleviated by the infusion of large amounts of plasma, the placenta failed as the result of numerous white infarcts. She delivered a 948 g live baby by cesarean section in the 33rd week. The baby had transient thrombocytopenia but did not suffer from TTP. The mother required plasma infusions every 3-4 weeks for about five months, and she has continued in remission.


Development and testing of a field diagnostic assay for peste des petits ruminants virus.
We have developed an immunochromatographic test for the diagnosis of peste des petits ruminants (PPR) under field conditions. The diagnostic assay has been tested in the laboratory and also under field conditions in Ivory Coast, Pakistan, Ethiopia and Uganda. The test is carried out on a superficial swab sample (ocular or nasal) and showed a sensitivity of 84% relative to PCR. The specificity was 95% over all nasal and ocular samples. The test detected as little as 10(3) TCID50 (50% tissue culture infectious doses) of cell culture-grown virus, and detected virus isolates representing all four known genetic lineages of peste des petits ruminants virus. Virus could be detected in swabs from animals as early as 4 days post-infection, at a time when clinical signs were minimal. Feedback from field trials was uniformly positive, suggesting that this diagnostic tool may be useful for current efforts to control the spread of PPR.


Factors Associated With the Outcome of a First-Line Intervention for Patients With Hip or Knee Osteoarthritis or Both: Data From the BOA Register.
This study explored the association of patients' demographics, health status, symptom severity, previous osteoarthritis (OA) care, and psychological status with the change in pain severity following a first-line intervention including education and exercise for OA provided nationwide in Swedish primary care. This register-based cohort study included 23,309 people with knee or hip OA from the Better Management of Patients with OA register. Linear regression models were used to assess the association of independent variables with the change in pain from baseline to 3 and 12 months. All the analyses were stratified based on the affected joint (hip vs knee). In people with hip and people with knee OA, high levels of baseline pain were associated with decreased pain at both follow-ups (3 months: knee B = -.67; hip B = -.64; 12 months: knee B = -.70; hip B = -.66), whereas being older, overweight, or female had a weak or no association. Finally, at both follow-ups, bilateral OA was associated with increased pain only in people with knee OA, whereas comorbidities and the willingness to undergo surgery were associated with increased pain regardless of the affected joint. Baseline pain showed the strongest association among the analyzed variables, whereas sex, age, and body mass index appear to be weakly associated with the pain change after a first-line intervention. Comorbidities and willingness to undergo surgery showed a potentially important association and may have a negative impact on the pain change following a first-line intervention. In people with hip or knee OA, age, sex, body mass index, and previous surgery are only weakly associated with the change in pain after a first-line intervention supporting the evidence recommending exercise and education as a foundation for all OA therapy. Having comorbidities and being willing to undergo surgery is associated with a worse outcome from a first-line intervention, including exercise and education. Individualized treatments addressing the disease perception and the specific comorbidity profile may improve the outcomes.


Spinal Cord Injury Patients Exhibit Changes in Motor-Related Activity and Topographic Distribution.
Spinal Cord Injury (SCI) is a common disease that usually limits the patient's independence by affecting their motor function. SCI patients usually present neuroplasticity, which allows brain signals transmission through spread pathways. Some innovative rehabilitation therapies, such as functional electrical stimulation (FES) or Brain-computer interfaces (BCIs) jointly with motor neuroprostheses, provide hope for functional restoration. BCIs require the analysis of event-related EEG potentials (ERPs). Movement-related cortical potentials (MRCPs) and event-related desynchroni-zation and synchronization (ERD/ERS) are the most commonly studied ERPs during motor activity. ERPs of healthy subjects may vary from SCI patients. Thus, this study aimed to compare ERPs between healthy subjects and SCI patients during upper-limb movements (forearm supination and pronation, and hand open). Differences between controls and SCI patients were shown in terms of ERPs' amplitude as well as in topographic maps. Changes in amplitude were more substantial in ERD potentials than in MRCPs, while topographic maps showed better localization of all features in healthy patients. The level of SCI injury determines the patients' mobility. A comparison between complete, partial and no motor function subjects showed lower values of feature's amplitudes in the latter group.Clinical Relevance- This demonstrates the existence of significant statistical differences between healthy and SCI subjects, and might be helpful when performing SCI rehabilitation techniques such as designing BCI and neuroprostheses, or analyzing and understanding the brain plasticity process.


Vitamin D3-implications for brain development.
There is growing evidence that 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) is active in the brain but until recently there was a lack of evidence about its role during brain development. Guided by certain features of the epidemiology of schizophrenia, our group has explored the role of 1,25(OH)(2)D(3) in brain development using whole animal models and in vitro culture studies. The expression of the vitamin D receptor (VDR) in the embryonic rat brain rises steadily between embryonic day 15-23, and 1,25(OH)(2)D(3) induces the expression of nerve growth factor and stimulates neurite outgrowth in embryonic hippocampal explant cultures. In the neonatal rat, low prenatal vitamin D(3) in utero leads to increased brain size, altered brain shape, enlarged ventricles, reduced expression of nerve growth factors, reduced expression of the low affinity p75 receptor and increased cellular proliferation. In summary, there is growing evidence that low prenatal levels of 1,25(OH)(2)D(3) can influence critical components of orderly brain development. It remains to be seen if these processes are of clinical relevance in humans, but in light of the high rates of hypovitaminosis D in pregnant women and neonates, this area warrants further scrutiny.


Delayed cardiac tamponade after blunt chest trauma: case report.
A case of cardiac tamponade without evidence of hemopericardium within 2 weeks after blunt chest trauma is reported. The cause was a pericardial effusion that may have resulted from the post-cardiac injury syndrome. Pericardiotomy drained 600 mL of straw-colored clear fluid. A high index of suspicion for cardiac tamponade must be maintained in such patients for weeks after injury.


Cloning and characterization of glutamate receptors in Californian sea lions (Zalophus californianus).
Domoic acid produced by marine algae has been shown to cause acute and chronic neurologic sequelae in Californian sea lions following acute or low-dose exposure. Histological findings in affected animals included a degenerative cardiomyopathy that was hypothesized to be caused by over-excitation of the glutamate receptors (GluRs) speculated to be present in the sea lion heart. Thus tissues from five sea lions without lesions associated with domoic acid toxicity and one animal with domoic acid-induced chronic neurologic sequelae and degenerative cardiomyopathy were examined for the presence of GluRs. Immunohistochemistry localized mGluR 2/3, mGluR 5, GluR 2/3 and NMDAR 1 in structures of the conducting system and blood vessels. NMDAR 1 and GluR 2/3 were the most widespread as immunoreactivity was observed within sea lion conducting system structures. PCR analysis, cloning and subsequent sequencing of the seal lion GluRs showed only 80% homology to those from rats, but more than 95% homologous to those from dogs. The cellular distribution and expression of subtypes of GluRs in the sea lion hearts suggests that exposure to domoic acid may induce cardiac damage and functional disturbances.


Do genetic make-up and growth manipulation affect tomato fruit size by cell number, or cell size and DNA endoreduplication?
This work investigated the link between genetic and developmental controls of fruit size and composition. On two isogenic lines (CF12-C and CF14-L), differing by fruit weight and sugar content quantitative trait loci (QTLs) identified previously, basal and tip fruits were characterized at anthesis and at maturity through their growth, dry matter and sugar content, number and size of cells and nuclei DNA content. The influence of competition was assessed by removing either basal or tip ovaries at anthesis. On an intact inflorescence, CF12-C fruits grew less than CF14-L fruits, with 1.67 fewer cell layers and similar cell size, suggesting that genes controlling cell division may be responsible for this fruit size variation. Truss thinning masked the QTL effect on fruit size, mainly by reducing the difference in cell number between the two lines and by promoting cell expansion in tip fruits, so that fruit growth was similar at both positions and for both lines. Thus, in these lines, cell number exerts a control on final fruit size only when there is competition among fruits. Different responses of basal and tip fruits after flower removal suggested that this treatment induced changes in hormonal relationships within the truss. No fixed relationship between DNA endoreduplication and cell size was found, as while cell size and dry matter and sugar contents differed with tomato lines, fruit position and truss size, endoreduplication patterns were the same. CF12-C fruits had a higher dry matter (+0.3% of fresh weight) and carbohydrates (+8% of dry matter) content than CF14-L fruits. The percentage dry matter was independent of truss size but decreased slightly from basal to tip fruits.


An in vitro study of the antimicrobial activity of some endodontic medicaments against Enteroccus faecalis biofilms.
The in vitro antimicrobial activity of a series of endodontic medicaments and their bases against biofilms of Enterococcus faecalis was investigated. The medicaments tested were Pulpdent paste, Ledermix paste, a 50:50 Ledermix and Pulpdent mixture, and a replica of Ledermix paste. Bases included methyl cellulose with water, polyethylene glycol (PEG), and PEG with zinc oxide, calcium chloride and the other components (inactives) that make up the Ledermix paste base. Biofilms grown on cellulose nitrate membrane filters were exposed to the medicaments for up to 5 days. The number of surviving colony forming units (CFU) was determined at days 1, 3 and 5. The results were expressed as a bacterial survival index (BSI) when compared to the unexposed control. Pulpdent produced the greatest reduction of BSI, followed by the 50:50 mixture of Pulpdent and Ledermix pastes. Ledermix paste, its replica and the individual bases showed no significant reductions in the BSI for E. faecalis. Within the limitations of this laboratory study, calcium hydroxide containing preparations had greater potential for reducing the survival of E. faecalis in a biofilm environment.


Visit-to-visit blood pressure variability is a strong predictor of cardiovascular events in hemodialysis: insights from FOSIDIAL.
Optimal blood pressure (BP) targets are still controversial in end-stage renal disease. Recent data have highlighted shortcomings of the usual BP hypothesis in other patient populations and emphasized the importance of visit-to-visit variability of BP in predicting cardiovascular events. The Fosinopril in Dialysis Study failed to demonstrate the efficacy of 2-year angiotensin-converting enzyme inhibition with fosinopril versus placebo in 397 hemodialysis patients with left ventricular hypertrophy but provided an opportunity to assess the influence of BP variability on cardiovascular events. The primary end point was the occurrence of a composite of cardiovascular death, nonfatal myocardial infarction, unstable angina, stroke, revascularization, hospitalization for heart failure, and resuscitated cardiac arrest. The variations in BP throughout the 17 visits were assessed by within-patient overall variability of systolic, diastolic, and pulse pressures between adjacent readings, by within-patient overall variability of systolic/diastolic/pulse pressures, and the residual of the linear fit. Compared with our previous predictive model of cardiovascular events occurrence based on stroke, peripheral arterial disease, coronary artery disease, diabetes mellitus, left ventricular mass, and age (which exhibited similar coefficients herein), the percentage of explained variance improved by 30.1% (R(2)=0.141-0.183) when adding the coefficient of variation of within-patient overall variability of systolic BP. Usual BP parameters were neither cardiovascular events predictors nor correlated to BP variability. Visit-to-visit BP variability was extremely high in hemodialysis patients compared with other populations and a major determinant of cardiovascular events. Such assessments should be prioritized for testing prevention strategies in end-stage renal disease.


Endothelial TLR4 Expression Mediates Vaso-Occlusive Crisis in Sickle Cell Disease.
Heme, released from red blood cells in sickle cell disease (SCD), interacts with toll-like receptor 4 (TLR4) to activate NF-κB leading to the production of cytokines and adhesion molecules which promote inflammation, pain, and vaso-occlusion. In SCD, TLR4 inhibition has been shown to modulate heme-induced microvascular stasis and lung injury. We sought to delineate the role of endothelial verses hematopoietic TLR4 in SCD by developing a TLR4 null transgenic sickle mouse. We bred a global Tlr4-/- deficiency state into Townes-AA mice expressing normal human adult hemoglobin A and Townes-SS mice expressing sickle hemoglobin S. SS-Tlr4-/- had similar complete blood counts and serum chemistries as SS-Tlr4+/+ mice. However, SS-Tlr4-/- mice developed significantly less microvascular stasis in dorsal skin fold chambers than SS-Tlr4+/+ mice in response to challenges with heme, lipopolysaccharide (LPS), and hypoxia/reoxygenation (H/R). To define a potential mechanism for decreased microvascular stasis in SS-Tlr4-/- mice, we measured pro-inflammatory NF-κB and adhesion molecules in livers post-heme challenge. Compared to heme-challenged SS-Tlr4+/+ livers, SS-Tlr4-/- livers had lower adhesion molecule and cytokine mRNAs, NF-κB phospho-p65, and adhesion molecule protein expression. Furthermore, lung P-selectin and von Willebrand factor immunostaining was reduced. Next, to establish if endothelial or hematopoietic cell TLR4 signaling is critical to vaso-occlusive physiology, we created chimeric mice by transplanting SS-Tlr4-/- or SS-Tlr4+/+ bone marrow into AA-Tlr4-/- or AA-Tlr4+/+ recipients. Hemin-stimulated microvascular stasis was significantly decreased when the recipient was AA-Tlr4-/- . These data demonstrate that endothelial, but not hematopoietic, TLR4 expression is necessary to initiate vaso-occlusive physiology in SS mice.


A multidisciplinary approach to designing and evaluating Electronic Medical Record portal messages that support patient self-care.
We describe a project intended to improve the use of Electronic Medical Record (EMR) patient portal information by older adults with diverse numeracy and literacy abilities, so that portals can better support patient-centered care. Patient portals are intended to bridge patients and providers by ensuring patients have continuous access to their health information and services. However, they are underutilized, especially by older adults with low health literacy, because they often function more as information repositories than as tools to engage patients. We outline an interdisciplinary approach to designing and evaluating portal-based messages that convey clinical test results so as to support patient-centered care. We first describe a theory-based framework for designing effective messages for patients. This involves analyzing shortcomings of the standard portal message format (presenting numerical test results with little context to guide comprehension) and developing verbally, graphically, video- and computer agent-based formats that enhance context. The framework encompasses theories from cognitive and behavioral science (health literacy, fuzzy trace memory, behavior change) as well as computational/engineering approaches (e.g., image and speech processing models). We then describe an approach to evaluating whether the formats improve comprehension of and responses to the messages about test results, focusing on our methods. The approach combines quantitative (e.g., response accuracy, Likert scale responses) and qualitative (interview) measures, as well as experimental and individual difference methods in order to investigate which formats are more effective, and whether some formats benefit some types of patients more than others. We also report the results of two pilot studies conducted as part of developing the message formats.


SMG1 acts as a novel potential tumor suppressor with epigenetic inactivation in acute myeloid leukemia.
Suppressor with morphogenetic effect on genitalia family member (SMG1) belongs to a family of phosphoinositide 3-kinase-related kinases and is the main kinase involved in nonsense-mediated mRNA decay. Recently, SMG1 was suggested as a novel potential tumor suppressor gene, particularly in hypoxic tumors. To investigate the function of SMG1 in acute myeloid leukemia (AML), we performed methylation-specific polymerase chain reaction and found that SMG1 was hypermethylated in the promoter region. SMG1 hypermethylation was found in 66% (33/50) of AML samples compared with none (0/14) of the normal controls. SMG1 mRNA was down-regulated in AML patients with hypermethylation status whereas it was readily expressed in patients without methylation. Moreover, treatment of AML cells with demethylating agent 5-aza-2'-deoxycytidine (decitabine) inhibited AML cell growth and induced apoptosis by reversing SMG1 methylation status and restoring SMG1 expression. On the other hand, knockdown of SMG1 by RNA interference inhibited apoptosis. We also found that mTOR expression level was negatively correlated to SMG1 expression in AML patients which indicated that SMG1 and mTOR maybe act antagonistically to regulate AML cell growth. In conclusion, our results indicate that SMG1 acts as a potential tumor suppressor with epigenetic regulation in AML.


Contribution of Pulmonary CYP-mediated Bioactivation of Naphthalene to Airway Epithelial Injury in the Lung.
Previous studies have established that cytochrome P450 enzymes (CYPs) in both liver and lung are capable of bioactivating naphthalene (NA), an omnipresent air pollutant and possible human carcinogen, in vitro and in vivo. The aim of this study was to examine the specific contribution of pulmonary CYPs in airway epithelial cells to NA-induced airway toxicity. We used a lung-Cpr-null mouse model, which undergoes doxycycline-induced, Cre-mediated deletion of the Cpr (a redox partner of all microsomal CYPs) gene specifically in airway epithelial cells. In 2-month-old lung-Cpr-null mice, Cpr deletion occurred in 75%-82% of epithelial cells of conducting airways. The extent of NA-induced acute lung toxicity (as indicated by total protein concentration and lactate dehydrogenase activity in bronchoalveolar lavage fluid collected at 24-h after initiation of a 4-h, nose-only, 10-ppm NA inhalation exposure) was substantially lower (by 37%-39%) in lung-Cpr-null mice, compared with control littermates. Moreover, the extent of cellular proliferation (as indicated by 5-bromo-2'-deoxyuridine incorporation) was noticeably lower in both proximal and distal airways (by 59% and 65%, respectively) of NA-treated lung-Cpr-null mice, compared with control littermates, at 2-day post-NA inhalation exposure. A similar genotype-related difference in the extent of postexposure cell proliferation was also observed in mice exposed to NA via intraperitoneal injection at 200 mg/kg. These results directly validate the hypothesis that microsomal CYP enzymes in airway epithelial cells play a large role in causing injury to airway epithelia following exposure to NA via either inhalation or intraperitoneal route.


Ribonucleotide reductase as a therapeutic target for drug repurposing as anthelmintics.
Visceral cestodiases, like echinococcoses and cysticercoses, are zoonoses of worldwide distribution and are responsible for public health problems in many countries, especially in underdeveloped regions. Current treatments have low efficiency and there are few drugs currently in use for chemotherapy, making the development of new anthelmintics an urgent matter. The nucleotide salvage pathways are the only ones available for nucleotide synthesis in cestodes and other parasitic helminths, and, here, we used in silico approaches to assess the potential of the enzymes in these pathways as targets for drug repurposing as anthelminthics. First, a genomic survey allowed to identify a repertoire of 28 enzymes of the purine and pyrimidine salvage pathways from the cestode Echinococcus granulosus sensu stricto. Regarding purines, the parasite relies on salvaging free bases rather than salvaging nucleosides. Pyrimidines, on the other hand, can be salvaged from both bases and nucleosides. Druggability of the parasite enzymes was assessed, as well as the availability of commercial inhibitors for them. Druggable enzymes were then ranked according to their potential for drug repurposing and the 17 most promising enzymes were selected for evolutionary analyses. The constructed phylogenetic trees allowed to assess the degree of conservation among ortholog enzymes from parasitic helminths and their mammalian hosts. Positive selection is absent in all assessed flatworm enzymes. A potential target enzyme for drug repurposing, ribonucleotide reductase (RNR), was selected for further assessment. RNR 3D-modelling showed structural similarities between the E. granulosus and the human orthologs suggesting that inhibitors of the human RNR should be effective against the E. granulosus enzyme. In line with that, E. granulosus protoscolices treated in vitro with the inhibitor hydroxyurea had their viability and DNA synthesis reduced. These results are consistent with nucleotide synthesis inhibition and confirm the potential of a nucleotide salvage inhibitors for repurposing as an anthelmintic.


Liver mTOR controls IGF-I bioavailability by regulation of protein kinase CK2 and IGFBP-1 phosphorylation in fetal growth restriction.
Fetal growth restriction (FGR) increases the risk for perinatal complications and predisposes the infant to diabetes and cardiovascular disease later in life. No treatment for FGR is available, and the underlying pathophysiology remains poorly understood. Increased IGFBP-1 phosphorylation has been implicated as an important mechanism by which fetal growth is reduced. However, to what extent circulating IGFBP-1 is phosphorylated in FGR is unknown, and the molecular mechanisms linking FGR to IGFBP-1 phosphorylation have not been established. We used umbilical cord plasma of appropriate for gestational age (AGA) and growth-restricted human fetuses and determined IGFBP-1 and IGF-I concentrations (ELISA) and site-specific IGFBP-1 phosphorylation (Western blotting using IGFBP-1 phospho-site specific antibodies). In addition, we used a baboon model of FGR produced by 30% maternal nutrient restriction and determined mammalian target of rapamycin (mTOR)C1 activity, CK2 expression/activity, IGFBP-1 expression and phosphorylation, and IGF-I levels in baboon fetal liver by Western blot, enzymatic assay, and ELISA. HepG2 cells and primary fetal baboon hepatocytes were used to explore mechanistic links between mTORC1 signaling and IGFBP-1 phosphorylation. IGFBP-1 was hyperphosphorylated at Ser101, Ser119, and Ser169 in umbilical plasma of human FGR fetuses. IGFBP-1 was also hyperphosphorylated at Ser101, Ser119, and Ser169 in the liver of growth-restricted baboon fetus. mTOR signaling was markedly inhibited, whereas expression and activity of CK2 was increased in growth-restricted baboon fetal liver in vivo. Using HepG2 cells and primary fetal baboon hepatocytes, we established a mechanistic link between mTOR inhibition, CK2 activation, IGFBP-1 hyperphosphorylation, and decreased IGF-I-induced IGF-I receptor autophosphorylation. We provide clear evidence for IGFBP-1 hyperphosphorylation in FGR and identified an mTOR and CK2-mediated mechanism for regulation of IGF-I bioavailability. Our findings are consistent with the model that inhibition of mTOR in the fetal liver, resulting in increased CK2 activity and IGFBP-1 hyperphosphorylation, constitutes a novel mechanistic link between nutrient deprivation and restricted fetal growth.


Assessing contamination and compliance in the prostate component of the Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening Trial.
Recently, the Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening Trial published 7-year complete prostate cancer mortality results, which showed no benefit of screening with prostate specific antigen (PSA) and digital rectal examination (DRE). An issue of concern was the substantial level of 'contamination', or use of PSA and DRE in control arm men. To provide a detailed description of contamination in PLCO. Surveys inquiring about the most recent PSA and DRE use were given to a sample of control arm men throughout the screening phase of PLCO (years 0-5). A probability model was utilized to translate survey results into actual frequency counts of tests. To assess the impact of contamination, Surveillance, Epidemiology, and End (SEER) incidence rates from the pre-screening era (1985-1987) as well as contemporaneous rates, were applied to PLCO person-years of observation. Results Of 38,350 control arm men, 2427 were surveyed. Pre-trial screening and college education were statistically significantly associated with increased contamination rates. The estimated mean number of screening PSAs (DREs) in the control arm was 2.7 (1.1); this compares to 5.0 (3.5) in the screened arm. 1984 and 2538 prostate cancers were observed in the control and screened arms, respectively, during the screening phase. In the absence of screening, 960 and 949 would have been expected; with contemporaneous incidence rates, 1630 and 1611 were expected. Due to the limitations of the surveys, in terms of both reach and scope, the exact level of PSA and DRE use in control arm men cannot be known. Use of prostate screening by control arm men was substantial, but also substantially less than in screened arm men. Detailed quantitative analyses of screening use across arms are critical for understanding current and future findings from the prostate component of PLCO.


Comparison of core-genome MLST, coreSNP and PFGE methods for Klebsiella pneumoniae cluster analysis.
In this work we compared the most frequently used Klebsiella pneumoniae typing methods: PFGE, cgMLST and coreSNP. We evaluated the discriminatory power of the three methods to confirm or exclude nosocomial transmission on K. pneumoniae strains isolated from January to December 2017, in the framework of the routine surveillance for multidrug-resistant organisms at the San Raffaele Hospital, in Milan. We compared the results of the different methods to the results of epidemiological investigation. Our results showed that cgMLST and coreSNP are more discriminant than PFGE, and that both approaches are suitable for transmission analyses. cgMLST appeared to be inferior to coreSNP in the K. pneumoniae CG258 phylogenetic reconstruction. Indeed, we found that the phylogenetic reconstruction based on cgMLST genes wrongly clustered ST258 clade1 and clade2 strains, conversely properly assigned by coreSNP approach. In conclusion, this study provides evidences supporting the reliability of both cgMLST and coreSNP for hospital surveillance programs and highlights the limits of cgMLST scheme genes for phylogenetic reconstructions.


Legacy of a Butterfly's Parental Microbiome in Offspring Performance.
An insect's phenotype can be influenced by the experiences of the parental generation. However, the effects of the parental symbiotic microbiome and host plant use on the offspring are unclear. We addressed this gap of knowledge by studying Pieris brassicae, a multivoltine butterfly species feeding on different brassicaceous plants across generations. We investigated how disturbance of the parental bacterial community by antibiotic treatment affects F1 larval traits. We tested the effects depending on whether F1 larvae are feeding on the same plant species as their parents or on a different one. The parental treatment alone had no impact on the biomass of F1 larvae feeding on the parental plant species. However, the parental treatment had a detrimental effect on F1 larval biomass when F1 larvae had a different host plant than their parents. This effect was linked to higher larval prophenoloxidase activity and greater downregulation of the major allergen gene (MA), a glucosinolate detoxification gene of P. brassicae Bacterial abundance in untreated adult parents was high, while it was very low in F1 larvae from either parental type, and thus unlikely to directly influence larval traits. Our results suggest that transgenerational effects of the parental microbiome on the offspring's phenotype become evident when the offspring is exposed to a transgenerational host plant shift.IMPORTANCE Resident bacterial communities are almost absent in larvae of butterflies and thus are unlikely to affect their host. In contrast, adult butterflies contain conspicuous amounts of bacteria. While the host plant and immune state of adult parental butterflies are known to affect offspring traits, it has been unclear whether also the parental microbiome imposes direct effects on the offspring. Here, we show that disturbance of the bacterial community in parental butterflies by an antibiotic treatment has a detrimental effect on those offspring larvae feeding on a different host plant than their parents. Hence, the study indicates that disturbance of an insect's parental microbiome by an antibiotic treatment shapes how the offspring individuals can adjust themselves to a novel host plant.


Preparation and characterization of eudragit retard nanosuspensions for the ocular delivery of cloricromene.
The purpose of this study was to improve the stability of cloricromene (AD6) in ophthalmic formulations and its drug availability at the ocular level. To this end, AD6-loaded polymeric nanoparticle suspensions were made using inert polymer resins (Eudragit RS100 and RL100). We modified the quasi-emulsion solvent diffusion technique by varying some formulation parameters (the drug-to-polymer ratio, the total drug and polymer amount, and the stirring speed). The chemical stability of AD6 in the nanosuspensions was assessed by preparing some formulations using (unbuffered) isotonic saline or a pH 7 phosphate buffer solution as the dispersing medium. The formulations were stored at 4 degrees C, and the rate of degradation of AD6 was followed by high performance liquid chromatography (HPLC). The obtained nanosuspensions showed mean sizes and a positive surface charge (zeta-potential) that make them suitable for an ophthalmic application; these properties were maintained upon storage at 4 degrees C for several months. In vitro dissolution tests confirmed a modified release of the drug from the polymer matrixes. Nanosuspensions prepared with saline solution and no or lower amounts of surfactant (Tween 80) showed an enhanced stability of the ester drug for several months, with respect to an AD6 aqueous solution. Based on the technological results, AD6-loaded Eudragit Retard nanoparticle suspensions appear to offer promise as a means to improving the shelf life and bioavailability of this drug after ophthalmic application.


Estimated burden of fungal infections in Kenya.
Kenya is a developing country with a high rate of tuberculosis (TB) and a moderate HIV infection burden. No estimate of the burden of fungal diseases in Kenya is published. We used specific populations at risk and fungal infection frequencies from the literature to estimate national incidence or prevalence of serious fungal infections. Used sources were: 2010 WHO TB statistics, Kenya Acquired Immunodeficiency Syndrome (AIDS) Epidemic Update 2012, Kenya Facts and figures 2012, Kenya Demographic and Health Survey 2008-2009. Of Kenya's population of ~40 million, 43% are under 15 years old and approximately 594,660 Kenyan women get >4 episodes Candida vulvovaginitis annually (2,988/100,000). The HIV/AIDS population at risk of opportunistic infections (OI) is 480,000 and the OI estimates include 306,000 patients with oral thrush (768/100,000), 114,000 with oesophageal candidiasis (286/100,000), 11,900 with cryptococcal meningitis (29/100,000) and 17,000 patients with Pneumocystis pneumonia (42/100,000). Chronic pulmonary aspergillosis following TB has a prevalence of 10,848 cases (32/100,000). The adult asthma prevalence is 3.1% and assuming 2.5% have allergic bronchopulmonary aspergillosis then 17,696 (44/100,000) are affected. Invasive aspergillosis, candidaemia and Candida peritonitis are probably uncommon. Tinea capitis infects 9.6% of children in Kenya, while fungal keratitis and otomycoses are difficult to estimate. At any one time, about 7% of the Kenyan population suffers from a significant fungal infection, with recurrent vaginitis and tinea capitis accounting for 82% of the infections. These estimates require further epidemiological studies for validation.


The natural history and treatment outcome of blast phase BCR-ABL- myeloproliferative neoplasms.
We analyzed the outcomes of 74 patients diagnosed with BCR-ABL(-) myeloproliferative neoplasms in blast phase receiving induction chemotherapy (55%), low-intensity therapy (16%), stem cell transplantation (SCT; 3%), or supportive care (26%). Median survival from the date of blastic transformation was 5 months. Patients receiving supportive therapy had a median survival of 6 weeks. Complete remission with or without blood recovery was achieved in 46% of patients receiving induction chemotherapy, but remissions were not durable with a median progression-free survival of only 5 months. Eight patients received SCT either as first therapy or after responding to antileukemia therapy. These patients had a markedly superior survival, with 73% alive at a median follow-up of 31 months. JAK2V617F kinetics were assessed in 16 patients: 0 of 4 negative patients became positive at transformation, and among 12 positive patients, 1 had an increase in JAK2V617F% at transformation, 7 had a substantial decrease, and 4 had stable levels. Myeloproliferative neoplasm blast phase is associated with a dismal prognosis. Responses to chemotherapy can be achieved but are not durable. Long-term survivors had all received SCT either as first therapy or in first remission.


In vivo and in vitro function of the Escherichia coli periplasmic cysteine oxidoreductase DsbG.
We have characterized in vivo and in vitro the recently identified DsbG from Escherichia coli. In addition to sharing sequence homology with the thiol disulfide exchange protein DsbC, DsbG likewise was shown to form a stable periplasmic dimer, and it displays an equilibrium constant with glutathione comparable with DsbA and DsbC. DsbG was found to be expressed at approximately 25% the level of DsbC. In contrast to earlier results (Andersen, C. L., Matthey-Dupraz, A., Missiakas, D., and Raina, S. (1997) Mol. Microbiol. 26, 121-132), we showed that dsbG is not essential for growth and that dsbG null mutants display no defect in folding of multiple disulfide-containing heterologous proteins. Overexpression of DsbG, however, was able to restore the ability of dsbC mutants to express heterologous multidisulfide proteins, namely bovine pancreatic trypsin inhibitor, a protein with three disulfides, and to a lesser extent, mouse urokinase (12 disulfides). As in DsbC, the putative active site thiols in DsbG are completely reduced in vivo in a dsbD-dependent fashion, as would be expected if DsbG is acting as a disulfide isomerase or reductase. However, the latter is not likely because DsbG could not catalyze insulin reduction in vitro. Overall, our results indicate that DsbG functions primarily as a periplasmic disulfide isomerase with a narrower substrate specificity than DsbC.


Effect of Alcohol Withdrawl on Glutathione S-transferase, Total Antioxidant Capacity and Amylase in Blood and Saliva of Alcohol-Dependent Males.
Alcohol biomarkers help in the early detection of alcoholism and its complications. There is a paucity of studies in India on the salivary markers of systemic diseases in general and on salivary alcohol biomarkers in particular. The present study was aimed at assessing the effect of alcohol withdrawal on the antioxidants and amylase in blood and saliva, and at finding the correlation between the blood and the salivary parameters in alcoholics. Sixty alcohol-dependent males who were in the age group of 30 - 70 years, who were admitted to the Deaddiction Centre for alcohol withdrawal treatment for one month, were the subjects of this study; age-matched healthy individuals were the controls. In the blood and saliva samples, the activities of Glutathione S-Transferase (GST) and amylase and the Total Antioxidant Capacity (TAC) were assayed. The alcohol-dependent subjects showed significantly lower GST and amylase activities and the TAC in blood and saliva as compared to those in the controls (P<0.001). The alcohol withdrawal caused a significant increase in the GST and amylase activities and the TAC to near-control values. In the alcohol-dependent subjects, there was a significant correlation between the values in blood and saliva with respect to GST and TAC. Alcoholism causes an impaired antioxidant capacity and a decreased secretion of amylase, which is ameliorated due to the alcohol withdrawal regimen . The strong correlation between blood and saliva with respect to the antioxidants suggests the potential future use of saliva as a laboratory tool in clinical medicine.


Cost sensitive hierarchical document classification to triage PubMed abstracts for manual curation.
The Immune Epitope Database (IEDB) project manually curates information from published journal articles that describe immune epitopes derived from a wide variety of organisms and associated with different diseases. In the past, abstracts of scientific articles were retrieved by broad keyword queries of PubMed, and were classified as relevant (curatable) or irrelevant (not curatable) to the scope of the database by a Naïve Bayes classifier. The curatable abstracts were subsequently manually classified into categories corresponding to different disease domains. Over the past four years, we have examined how to further improve this approach in order to enhance classification performance and to reduce the need for manual intervention. Utilizing 89,884 abstracts classified by a domain expert as curatable or uncuratable, we found that a SVM classifier outperformed the previously used Naïve Bayes classifier for curatability predictions with an AUC of 0.899 and 0.854, respectively. Next, using a non-hierarchical and a hierarchical application of SVM classifiers trained on 22,833 curatable abstracts manually classified into three levels of disease specific categories we demonstrated that a hierarchical application of SVM classifiers outperformed non-hierarchical SVM classifiers for categorization. Finally, to optimize the hierarchical SVM classifiers' error profile for the curation process, cost sensitivity functions were developed to avoid serious misclassifications. We tested our design on a benchmark dataset of 1,388 references and achieved an overall category prediction accuracy of 94.4%, 93.9%, and 82.1% at the three levels of categorization, respectively. A hierarchical application of SVM algorithms with cost sensitive output weighting enabled high quality reference classification with few serious misclassifications. This enabled us to significantly reduce the manual component of abstract categorization. Our findings are relevant to other databases that are developing their own document classifier schema and the datasets we make available provide large scale real-life benchmark sets for method developers.


Discovery of lost diversity of paternal horse lineages using ancient DNA.
Modern domestic horses display abundant genetic diversity within female-inherited mitochondrial DNA, but practically no sequence diversity on the male-inherited Y chromosome. Several hypotheses have been proposed to explain this discrepancy, but can only be tested through knowledge of the diversity in both the ancestral (pre-domestication) maternal and paternal lineages. As wild horses are practically extinct, ancient DNA studies offer the only means to assess this ancestral diversity. Here we show considerable ancestral diversity in ancient male horses by sequencing 4 kb of Y chromosomal DNA from eight ancient wild horses and one 2,800-year-old domesticated horse. Both ancient and modern domestic horses form a separate branch from the ancient wild horses, with the Przewalski horse at its base. Our methodology establishes the feasibility of re-sequencing long ancient nuclear DNA fragments and demonstrates the power of ancient Y chromosome DNA sequence data to provide insights into the evolutionary history of populations.


[A case of parathyroid hormone-related peptide producing gallbladder carcinoma presenting humoral hypercalcemia of malignancy].
Humoral hypercalcemia of malignancy (HHM) in neoplastic syndrome has been most commonly reported in squamous cell carcinoma. Gallbladder carcinoma with HHM is uncommon. In this report, we describe a male case of gallbladder carcinoma with marked hypercalcemia and a high level of serum parathyroid hormone-related peptide (PTHrP). An immunohistochemical examination using PTHrP was also positive.


2-Methyloxolane as a Bio-Based Solvent for Green Extraction of Aromas from Hops (Humulus lupulus L.).
The potential of using the bio-based solvent 2-methyloxolane, also known as 2-methyltetrahydrofuran or 2-MeTHF, as an alternative to petroleum solvents such as hexane, was investigated for the extraction of volatile compounds from hop cones (Humulus lupulus L.). Lab scale extractions were coupled with in silico prediction of solutes solubility to assess the technical potential of this bio-based solvent. The predictive approach was performed using the simulation software COSMO-RS (conductor like screening model for real solvants) and showed that the 2-methyloxolane is as good as or better than hexane to solubilize the majority of aromas from hop cones. The experimental results indicated that the highest aroma yield was obtained with 2-methyloxolane with 20.2% while n-hexane was only able to extract 17.9%. The characterization of aromas extracted by the two solvents showed a similar composition, where lupulone was the main component followed by humulone. No selectivity of the solvents was observed for any of the major analytes. Finally, a sensory analysis was performed on the extracts, showing that both concretes using 2-methyloxolane and hexane have similar olfactory profiles. The results indicate that 2-methyloxolane could be a promising bio-based extraction solvent for hexane substitution.


Extraaxial ependymoma of the posterior fossa.
We report an unusual case of an extraaxial ependymoma of the posterior fossa in an adult. MR imaging showed a heterogeneously enhancing extraaxial mass with a cystic component. Ependymoma should be included in the differential diagnosis of uncommon extraaxial masses of the posterior fossa.


Self-Reported Accommodation Needs for Patients with Disabilities in Primary Care.
People with disabilities experience barriers to engaging with health care due to inaccessible social and physical environments at primary care clinics. Despite legal mandates, identification and provision of necessary accommodations for this population at primary care clinics are poor. The objective of this cross-sectional study was to assess patient-reported disability status and accommodation needs among patients at a primary care clinic. An electronic health record-based Disability and Accommodations Questionnaire assessing disability status, types, and accommodation needs was developed by subject matter experts at Michigan Medicine and the University of Michigan Council for Disability Concerns. The questionnaire underwent multiple rounds of reviews and revisions before its use in clinical settings. A paper-based questionnaire was administered to all patients presenting for a wellness-based visit at an academic health system primary care clinic in southeast Michigan. Data were collected between March 2022 and August 2022. Approximately 13% of the 541 patients self-reported a disability, with 54.2% indicating at least one needed accommodation. The most commonly reported disabilities were mental health and hearing-related disabilities, by 4.8% and 4.6% of patients, respectively. The most frequently requested accommodations were communication- or language-based (for example, presence of an American Sign Language interpreter, assistive listening devices), cognitive-based (for example, inclusion of a support person with care decisions), and mobility-based (for example, assistance with transfers). The Disability and Accommodations Questionnaire helped identify the presence of a disability, its types, and any requested accommodations requested at a primary care health center.


Culicoides midge bites modulate the host response and impact on bluetongue virus infection in sheep.
Many haematophagous insects produce factors that help their blood meal and coincidently favor pathogen transmission. However nothing is known about the ability of Culicoides midges to interfere with the infectivity of the viruses they transmit. Among these, Bluetongue Virus (BTV) induces a hemorrhagic fever- type disease and its recent emergence in Europe had a major economical impact. We observed that needle inoculation of BTV8 in the site of uninfected C. nubeculosus feeding reduced viraemia and clinical disease intensity compared to plain needle inoculation. The sheep that developed the highest local inflammatory reaction had the lowest viral load, suggesting that the inflammatory response to midge bites may participate in the individual sensitivity to BTV viraemia development. Conversely compared to needle inoculation, inoculation of BTV8 by infected C. nubeculosus bites promoted viraemia and clinical symptom expression, in association with delayed IFN- induced gene expression and retarded neutralizing antibody responses. The effects of uninfected and infected midge bites on BTV viraemia and on the host response indicate that BTV transmission by infected midges is the most reliable experimental method to study the physio-pathological events relevant to a natural infection and to pertinent vaccine evaluation in the target species. It also leads the way to identify the promoting viral infectivity factors of infected Culicoides in order to possibly develop new control strategies against BTV and other Culicoides transmitted viruses.


Periodontal diseases: microbiological considerations.
The location of plaque-associated gingivitis at the gingival portion of the tooth plays an essential role in its genesis. However, at times local and other host response modifying factors also have an influence. The pathogeny of periodontitis is more complex. The microorganisms that comprise subgingival plaque are capable of acting directly on periodontal tissues or of modifying the host response, whereas the participation of the plaque per se (normal, decreased, or increased) is as decisive as the action of the bacteria themselves in the emergence of the disease. Different types of periodontitis are associated with specific microorganisms. The most periodontopathogenic are A. actinomycetemcomitans, P. gingivalis, and T. forsythensis. Periodontitis as a whole, represent the source of complications such as root caries, endoperiodontal processes and periodontal abscesses. They are associated with various illnesses such as atherosclerosis, diabetes, and respiratory infections, amongst others, as well as pathological oral halitosis. The different modalities of PCR are particularly important in the microbiological diagnosis of periodontitis, although on the negative side of things, it must be pointed out that in vitro sensitivity studies cannot be performed using this technique. First line antibiotic treatment of periodontitis includes amoxicillin/ clavulanic acid, metronidazole (associated or not with amoxicillin) and clindamycin.


[Health status in the population using contraceptives].
In order to partially understand the health status of women under contraceptive therapy and their possible social hazard of it, a descriptive, prospective and transversal study was undertaken in a 500 individuals, ages 20-35 years old, who currently were regulation their fertility. Data were obtained by means of predetermined questions from subjects attending several clinics of the IMSS. Anyone refusing to participate was discharged questions. The analysis excluded 8/500 due to incomplete responses. In 334/492, hormonal practice was in use; the IUD met 36.6% and the minor groups of 4.4 and 2.8% the method chosen was surgical or natural respectively. The IUD had been used over three years in 60/187, while in 39/145 were on the pill the same period of time; only 23/100 preferred the injectables. No previous medical advice was carried out in order to prescribe the contraceptive and furthermore in 461/492 no laboratory clinical tests were required before. In only 22% of the population the Pap-smear had been taken two years before and in 89/492 was never considered. The contraceptive strategies was induce risks in the future with no solid etiology if a prescription is decided, before the health status is evaluated. It looks like that present report is the first description associating health and risks in a non-selected population under contraceptive methodology.


Drug testing with alternative matrices II. Mechanisms of cocaine and codeine deposition in hair.
A 10-week inpatient study was performed to evaluate cocaine, codeine, and metabolite disposition in biological matrices collected from volunteers. An initial report described drug disposition in plasma, sebum, and stratum corneum collected from five African-American males. This report focuses on drug disposition in hair and sweat collected from the same five subjects. Following a three-week washout period, three doses of cocaine HCl (75 mg/70 kg, subcutaneous) and three doses of codeine SO4 (60 mg/70 kg, oral) were administered on alternating days in week 4 (low-dose week). The same dosing sequence was repeated in week 8 with doubled doses (high-dose week). Hair was collected by shaving the entire scalp once each week. Hair from the anterior vertex was divided into two portions. One portion was washed with isopropanol and phosphate buffer; the other portion was not washed. Hair was enzymatically digested, samples were centrifuged, and the supernatant was collected. Sweat was collected periodically by placing PharmChek sweat patches on the torso. Drugs were extracted from sweat patches with methanol/0.2 M sodium acetate buffer (75:25, v/v). Supernatants from hair digests, hair washes, and sweat patch extracts were processed by solid-phase extraction followed by gas chromatography-mass spectrometry analysis for cocaine, codeine, 6-acetylmorphine, and metabolites. Cocaine and codeine were the primary analytes identified in sweat patches and hair. Drugs were detected in sweat within 8 h after dosing, and drug secretion primarily occurred within 24 h after dosing. No clear relationship was observed between dose and drug concentrations in sweat. Drug incorporation into hair appeared to be dose-dependent. Drugs were detected in hair within 1-3 days after the last drug administration; peak drug concentrations generally occurred in the following 1-2 weeks; thereafter, drug concentrations decreased. Solvent washes removed 50-55% of cocaine and codeine from hair collected 1-3 days after the last drug dose. These data may reflect removal of drug that was deposited by sweat shortly after dosing. Drug removed by washing hair collected 1-3 weeks after the last dose was minimal for cocaine but variable for codeine. Drug in these specimens was likely transferred from blood to germinative hair cells followed by emergence of drug in growing hair. These findings suggest that drug deposition in hair occurs by multiple mechanisms.


Hyperthermia and chronic pancerebellar syndrome after cocaine abuse.
We describe a patient who developed pancerebellar syndrome as a result of hyperthermia that developed after cocaine use. The patient had long-standing schizophrenia and had been taking risperidone for 2 years, without evidence of abnormal movements. A literature review revealed a marked similarity between cocaine and neuroleptics in their ability to cause hyperthermia. Based on our observations and the compatible evidence from the literature, we suggest that cocaine use may cause hyperthermia and result in chronic pancerebellar dysfunction.


Immunocytochemistry suggests that the prevalence of a sub-type of beta-proteins determines the hardness in the epidermis of the hard-shelled turtle.
The corneous layer of the epidermis in hard-shelled turtles largely derives from the accumulation of beta-proteins as indicated by microscopic, in situ hybridization, and immunocytochemical and Western blotting analysis. The expression of mRNAs of one of the most common type of beta-proteins shows higher expression in upper spinosus and pre-corneous keratinocytes of growing scutes. Two beta-proteins of 14-16 kDa, indicated as Tu2 and Tu17 and representing two subtypes of beta-proteins co-accumulate in the thick corneous layer of the epidermis in hard-shelled turtle. The two beta-proteins apparently mix in differentiating and mature corneocytes although Tu2 appears more prevalent than Tu17. The specific role of the different subtypes in the formation of the hard corneous material of the carapace and plastron is not clear. It is hypothesized that the relative amount of beta-proteins belonging to the two subclasses in relation to the alpha-keratin meshwork present in keratinocytes contributes to the formation of a variably resistant and inflexible corneous layer. Tu17 may have a more globular structure than Tu2 and is likely present in denser areas of the corneous layer containing also alpha-keratin. The increase of cysteine-glycine-rich beta-proteins in the matrix located among alpha-keratin filaments may allow the formation of a hard corneous material, probably through increase of cross-bridge formation and hydrophobicity.


Burn injury decreases the antinociceptive effects of opioids.
Burn victim patients are frequently prescribed opioids at doses that are significantly higher than standard analgesic dosing guidelines, and, even despite an escalation in opioid dosing, many continue to experience pain. Thus, the aim of this study was to determine the effect of burn injury on opioid antinociception. Mice were examined for their baseline pain sensitivity thresholds using the von Frey filaments test. Then, they were subjected to burn or sham injury to the dorsal surface of the hindpaw and treated orally with morphine, oxycodone, hydrocodone (20 or 40 mg/kg), or saline twice daily throughout the study. They were retested on days 4, 7, 11, 14, 21, and 28 following the burn injury. The antinociceptive effects of the various drugs were analyzed by computing the daily difference between pain sensitivity threshold scores (in g) before and after treatment. This study showed that burn injury decreases opioid antinociception potency. A marked reduction was observed in the antinociceptive effectiveness of all opioids, and for both doses, in the burn-injured versus the sham animals. These results suggest that burn trauma limits the ability of opioids to be effective in reducing pain.


Formation of prostacyclin-sensitive platelet aggregates in human whole blood in vitro. Part II. The occurrence of the phenomenon in males suffering from acute myocardial infarction.
The de-aggregatory effect of prostacyclin (PGI2) and the rate of spontaneous platelet aggregation (SPA) were studied in vitro in whole blood of 24 males with acute myocardial infarction (MI) and 18 males, patient controls (PC). The de-aggregatory effect of PGI2 and the rate of SPA (measured as a percentage of changes in free platelet number in whole blood) were higher (p less than 0.01) in MI than PC. The de-aggregatory effect of PGI2 in whole blood was higher (p less than 0.05) on the first day of MI than on day 14 following MI. The highest de-aggregatory effect of PGI2 was found in whole blood of patients with MI complicated by ventricular fibrillation. In neither of the groups did the de-aggregatory effect of PGI2 correlate with patients' age, haematocrit, erythrocyte and leucocyte counts, triglycerides, HDL, LDL or total cholesterol levels. In the MI group, de-aggregatory effect of PGI2 was correlated with free platelet concentration (r = -0.59, p less than 0.05), elevation of glutamic oxalacetic transaminase (r = 0.53, p less than 0.05) and creatinine phosphokinase (r = 0.69, p less than 0.001). The de-aggregatory effect of PGI2 in blood of patients with evolving MI did not differ from that in PC. It is concluded that the increased rate of SPA and formation of PGI2-sensitive platelet aggregates in vitro in whole blood of MI patients are secondary to myocardial necrosis.


Sustained hemodynamic and clinical effects of a new cardiotonic agent, WIN 47203, in patients with severe congestive heart failure.
The hemodynamic and clinical effects of WIN 47203, a newly synthesized noncatecholamine, nonglycosidic inotropic agent, were studied in 11 patients with severe chronic congestive heart failure. Intravenous WIN 47203 increased cardiac index from 1.93 +/- 0.36 to 2.87 +/- 0.45 l/min/m2 (p less than 0.001) and reduced pulmonary capillary wedge pressure from 27.0 +/- 8.4 to 16.3 +/- 6.1 mm Hg (p less than 0.001). Mean systemic arterial pressure decreased from 75.2 +/- 6.7 to 72.4 +/- 6.3 mm Hg (p less than 0.01) and systemic vascular resistance from 1591 +/- 397 to 1071 +/- 293 dyn-sec-cm5 (p less than 0.001); heart rate was unchanged. Oral WIN 47203 produced similar hemodynamic improvement. Hemodynamic monitoring of six consecutive doses did not demonstrate evidence for attenuation of effectiveness. Chronic therapy with WIN 47203 produced substantial symptomatic improvement and increased maximal oxygen uptake at 1 week. Patients were further improved after 4 weeks of WIN 47203, and maximal oxygen uptake increased from 9.0 +/- 1.9 to 11.6 +/- 2.5 ml/kg/min (p less than 0.01 vs control). No overt clinical or laboratory manifestations of toxicity were observed. Withdrawal of WIN 47203 in two patients in whom clinical benefit was not sustained resulted in clinical and hemodynamic deterioration, which was reversed by reinstitution of the drug. Therefore, this study demonstrates the acute and sustained cardiotonic efficacy of WIN 47203 in man. If long-term administration remains well tolerated and without side effects, this drug appears to be very promising for treatment of chronic severe congestive heart failure.