Effects of inhaled versus intravenous vasodilators in experimental pulmonary hypertension.
Inhaled nitric oxide (NO) causes selective pulmonary vasodilation and improves gas exchange in acute lung failure. In experimental pulmonary hypertension, we compared the influence of the aerosolized vasodilatory prostaglandins (PG) PGI2 and PGE1 on vascular tone and gas exchange to that of infused prostanoids (PGI2, PGE1) and inhaled NO. An increase of pulmonary artery pressure (Ppa) from 8 to approximately 34 mmHg was provoked by continuous infusion of U-46619 (thromboxane A2 (TxA2) analogue) in blood-free perfused rabbit lungs. This was accompanied by formation of moderate lung oedema and severe ventilation-perfusion (V'/Q') mismatch, with predominance of shunt flow (>50%, assessed by the multiple inert gas elimination technique). When standardized to reduce the Pps by approximately 10 mmHg, inhaled NO (200 ppm), aerosolized PGI2 (4 ng x kg(-1) x min(-1)) and nebulized PGE1 (8 ng x kg(-1) x min(-1)) all reduced both pre- and postcapillary vascular resistance, but did not affect formation of lung oedema. All inhalative agents improved the V'/Q' mismatch and reduced shunt flow, the rank order of this capacity being NO > PGI2 > PGE1. In contrast, lowering of Ppa by intravascular administration of PGI2 and PGE1 did not improve gas exchange. "Supratherapeutic" doses of inhaled vasodilators in control lungs (400 ppm NO, 30 ng x kg(-1) x min(-1) of PGI2 or PGE1) did not provoke vascular leakage or affect the physiological V'/Q' matching. We conclude that aerosolization of prostaglandins I2 and E1 is as effective as inhalation of nitric oxide in relieving pulmonary hypertension. When administered via this route instead of being infused intravascularly, the prostanoids are capable of improving ventilation-perfusion matching, suggesting selective vasodilation in well-ventilated lung areas.


A Unique cis-Encoded Small Noncoding RNA Is Regulating Legionella pneumophila Hfq Expression in a Life Cycle-Dependent Manner.
Legionella pneumophila is an environmental bacterium that parasitizes protozoa, but it may also infect humans, thereby causing a severe pneumonia called Legionnaires' disease. To cycle between the environment and a eukaryotic host, L. pneumophila is regulating the expression of virulence factors in a life cycle-dependent manner: replicating bacteria do not express virulence factors, whereas transmissive bacteria are highly motile and infective. Here we show that Hfq is an important regulator in this network. Hfq is highly expressed in transmissive bacteria but is expressed at very low levels in replicating bacteria. A L. pneumophila hfq deletion mutant exhibits reduced abilities to infect and multiply in Acanthamoeba castellanii at environmental temperatures. The life cycle-dependent regulation of Hfq expression depends on a unique cis-encoded small RNA named Anti-hfq that is transcribed antisense of the hfq transcript and overlaps its 5' untranslated region. The Anti-hfq sRNA is highly expressed only in replicating L. pneumophila where it regulates hfq expression through binding to the complementary regions of the hfq transcripts. This results in reduced Hfq protein levels in exponentially growing cells. Both the small noncoding RNA (sRNA) and hfq mRNA are bound and stabilized by the Hfq protein, likely leading to the cleavage of the RNA duplex by the endoribonuclease RNase III. In contrast, after the switch to transmissive bacteria, the sRNA is not expressed, allowing now an efficient expression of the hfq gene and consequently Hfq. Our results place Hfq and its newly identified sRNA anti-hfq in the center of the regulatory network governing L. pneumophila differentiation from nonvirulent to virulent bacteria. The abilities of L. pneumophila to replicate intracellularly and to cause disease depend on its capacity to adapt to different extra- and intracellular environmental conditions. Therefore, a timely and fine-tuned expression of virulence factors and adaptation traits is crucial. Yet, the regulatory circuits governing the life cycle of L. pneumophila from replicating to virulent bacteria are only partly uncovered. Here we show that the life cycle-dependent regulation of the RNA chaperone Hfq relies on a small regulatory RNA encoded antisense to the hfq-encoding gene through a base pairing mechanism. Furthermore, Hfq regulates its own expression in an autoregulatory loop. The discovery of this RNA regulatory mechanism in L. pneumophila is an important step forward in the understanding of how the switch from inoffensive, replicating to highly virulent, transmissive L. pneumophila is regulated.


Real-time slit scanning microscopy in the meridional plane.
The standard microscope architecture around which confocal microscopes are built imposes fundamental restrictions on the speed with which images (three-dimensional data sets) can be obtained. Commercially available slit scanning confocal microscopes are able to produce optically sectioned images at frame rates well in excess of 100 Hz. However only the focal (x-y) plane can be imaged at this speed. To image a volume specimen it is necessary to physically change the distance between the objective lens and the specimen. This refocusing process is often necessarily slow and represents a bottleneck to the speed of image acquisition. We describe the construction of a slit scanning confocal microscope based on what we know to be a novel microscope architecture, which permits images of other planes and, particular, the meridional (x-z) plane to be acquired in real time.


Expanding the boundaries of local similarity analysis.
Pairwise comparison of time series data for both local and time-lagged relationships is a computationally challenging problem relevant to many fields of inquiry. The Local Similarity Analysis (LSA) statistic identifies the existence of local and lagged relationships, but determining significance through a p-value has been algorithmically cumbersome due to an intensive permutation test, shuffling rows and columns and repeatedly calculating the statistic. Furthermore, this p-value is calculated with the assumption of normality -- a statistical luxury dissociated from most real world datasets. To improve the performance of LSA on big datasets, an asymptotic upper bound on the p-value calculation was derived without the assumption of normality. This change in the bound calculation markedly improved computational speed from O(pm²n) to O(m²n), where p is the number of permutations in a permutation test, m is the number of time series, and n is the length of each time series. The bounding process is implemented as a computationally efficient software package, FASTLSA, written in C and optimized for threading on multi-core computers, improving its practical computation time. We computationally compare our approach to previous implementations of LSA, demonstrate broad applicability by analyzing time series data from public health, microbial ecology, and social media, and visualize resulting networks using the Cytoscape software. The FASTLSA software package expands the boundaries of LSA allowing analysis on datasets with millions of co-varying time series. Mapping metadata onto force-directed graphs derived from FASTLSA allows investigators to view correlated cliques and explore previously unrecognized network relationships. The software is freely available for download at: http://www.cmde.science.ubc.ca/hallam/fastLSA/.


Arterial stiffness and pharmacological interventions--the TRanscend arterial stiffNess Substudy (TRANS study).
The degree of arterial stiffness is correlated with the risk of cardiovascular diseases and it is a powerful predictor for morbidity and mortality. Studies have shown that arterial stiffness reduction is associated with an improvement in survival. Reduction of arterial stiffness by pharmacological drugs varies according to the drugs and doses used and duration of treatment. This effect on the arteries differs among the various classes of drugs and among individual drugs in the same class. Quantification of the stiffness and other properties of the arterial wall can be used to monitor the responses to therapy in individuals with hypertension and other cardiovascular diseases. These measures can then be used as surrogate markers for the risk of clinical events. Inhibition of the renin-angiotensin system (RAS) is associated with an important decrease in cardiovascular risk. Findings from clinical trials support the hypothesis that the protective effects of RAS inhibition are partly independent from blood pressure reduction and related to several mechanisms including vascular protective effects. The aim of the TRanscend Arterial stiffNess Substudy (TRANS) is to assess the effect of an angiotensin II receptor blocker (ARB), telmisartan, on the arterial stiffness in a subgroup of patients from the Telmisartan Randomized Assessment Study in aCE iNtolerant subjects with cardiovascular Disease (TRANSCEND) trial. The TRANSCEND trial is an international, multicenter, randomized double blind placebo controlled trial oftelmisartan that enrolled patients at high risk for cardiovascular events. Some clinical baseline data of the TRANS substudy are reported. When completed, the results of the TRANS substudy will show whether the beneficial effects of treatment with telmisartan on cardiovascular outcome may be related to an improvement in arterial stiffness.


Dedifferentiated liposarcoma of the spermatic cord: a series of 42 cases.
Dedifferentiated liposarcoma (DDL) of the spermatic cord is rare, with limited data regarding morphology and prognosis. We analyzed the morphologic spectrum of DDL of the spermatic cord and its clinical significance in 42 patients from 1990 to 2014. The median age of the patients was 70.5 years (range: 43 to 90 y). The median tumor size was 10.9 cm (range: 3 to 30 cm). Six cases were low-grade DDL, 28 were high-grade DDL, and in 8 cases both coexisted. Five cases had osteosarcoma, and 2 had leiomyosarcoma differentiation. Low-grade DDL had a fibrosarcoma-like (n=11), myxofibrosarcoma-like (n=2), and fibrosarcoma and inflammatory-like (n=1) pattern. High-grade DDL had a fibrosarcoma-like (n=22), undifferentiated pleomorphic sarcoma-like (n=7), myxofibrosarcoma and undifferentiated pleomorphic sarcoma-like (n=4), fibrosarcoma and undifferentiated pleomorphic sarcoma-like (n=2), and myxofibrosarcoma-like (n=1) pattern. Resection margins were positive in 22, negative in 13, and unknown in 7 cases. MDM2 was positive in 24/24 cases. Two patients received neoadjuvant radiotherapy (1 with chemotherapy). Patients received postoperative radiation (n=13), reexcision (n=5) with radiation (n=3), chemotherapy (n=2), chemotherapy and radiation (n=1), no therapy (n=11), and unknown (n=6). Follow-up information was available in 31 patients (mean: 54.2 mo; median: 38.5 mo; range: 2 to 180 mo). Seven patients developed local recurrence. Two patients had systemic metastases and succumbed to disease. Recurrence developed at an average of 40.9 months (median: 24 mo; range: 12 to 100 mo) after resection. Average follow-up in patients without recurrence was 45.2 months (median: 25 mo; range: 2 to 180 mo) (P=0.87). Margin status, size, and grade did not correlate with recurrence (P>0.05). Thus, DDL of the spermatic cord includes a spectrum of morphologic patterns. Complete excision is difficult at first attempt. Local recurrence is common, and relatively few patients develop systemic disease.


Administration of Zinc Chelators Improves Survival of Mice Infected with Aspergillus fumigatus both in Monotherapy and in Combination with Caspofungin.
Aspergillus fumigatus can infect immunocompromised patients, leading to high mortality rates due to the lack of reliable treatment options. This pathogen requires uptake of zinc from host tissues in order to successfully grow and cause virulence. Reducing the availability of that micronutrient could help treat A. fumigatus infections. In this study, we examined the in vitro effects of seven chelators using a bioluminescent strain of A. fumigatus 1,10-Phenanthroline and N,N,N',N'-tetrakis(2-pyridylmethyl)ethane-1,2-diamine (TPEN) proved to be the chelators most effective at inhibiting fungal growth. Intraperitoneal administration of either phenanthroline or TPEN resulted in a significant improvement in survival and decrease of weight loss and fungal burden for immunosuppressed mice intranasally infected with A. fumigatus In vitro both chelators had an indifferent effect when employed in combination with caspofungin. The use of TPEN in combination with caspofungin also significantly increased survival compared to that when using these drugs individually. Our results suggest that zinc chelation may be a valid strategy for dealing with A. fumigatus infections and that both phenanthroline and TPEN could potentially be used either independently or in combination with caspofungin, indicating that their use in combination with other antifungal treatments might also be applicable.


Tofacitinib in kidney transplantation.
This review will discuss the mechanism of action and important kidney transplant clinical trial data for the small molecule Janus kinase (JAK) 3 inhibitor tofacitinib , formerly known as CP-690,550 and tasocitinib. Successful kidney transplantation requires adequate immunosuppression. Current maintenance immunosuppressive protocols which rely on calcineurin inhibitors have long-term nephrotoxicity and negative impact on cardiometabolic risk factors. JAKs are cytoplasmic tyrosine kinases that participate in the signaling of a broad range of cell surface receptors, particularly members of the cytokine receptor common gamma (cγ) chain family. JAK3 inhibition has immunosuppressive effects and treatment with tofacitinib in clinical trials has demonstrated efficacy in autoimmune disorders such as psoriasis and rheumatoid arthritis. Nonhuman primate models of renal transplantation demonstrated prolonged graft survival with tofacitinib compared to control. Renal transplant clinical trials in humans have demonstrated tofacitinib to be noninferior to cyclosporine in terms of rejection rates and graft survival. There was also a lower rate of new onset diabetes after transplant. However, there was a trend toward more infections, including cytomegalovirus and BK virus nephritis. Tofacitinib may be a promising alternative to calcineurin inhibitors. The optimal therapeutic window is still being determined.


Evaluation of the diagnostic accuracy of two point-of-care tests for COVID-19 when used in symptomatic patients in community settings in the UK primary care COVID diagnostic accuracy platform trial (RAPTOR-C19).
Point-of-care lateral flow device antigen testing has been used extensively to identify individuals with active SARS-CoV-2 infection in the community. This study aimed to evaluate the diagnostic accuracy of two point-of-care tests (POCTs) for SARS-CoV-2 in routine community care. Adults and children with symptoms consistent with suspected current COVID-19 infection were prospectively recruited from 19 UK general practices and two COVID-19 testing centres between October 2020 and October 2021. Participants were tested by trained healthcare workers using at least one of two index POCTs (Roche-branded SD Biosensor Standard™ Q SARS-CoV-2 Rapid Antigen Test and/or BD Veritor™ System for Rapid Detection of SARS-CoV-2). The reference standard was laboratory triplex reverse transcription quantitative PCR (RT-PCR) using a combined nasal/oropharyngeal swab. Diagnostic accuracy parameters were estimated, with 95% confidence intervals (CIs), overall, in relation to RT-PCR cycle threshold and in pre-specified subgroups. Of 663 participants included in the primary analysis, 39.2% (260/663, 95% CI 35.5% to 43.0%) had a positive RT-PCR result. The SD Biosensor POCT had sensitivity 84.0% (178/212, 78.3% to 88.6%) and specificity 98.5% (328/333, 96.5% to 99.5%), and the BD Veritor POCT had sensitivity 76.5% (127/166, 69.3% to 82.7%) and specificity 98.8% (249/252, 96.6% to 99.8%) compared with RT-PCR. Sensitivity of both devices dropped substantially at cycle thresholds ≥30 and in participants more than 7 days after onset of symptoms. Both POCTs assessed exceed the Medicines and Healthcare products Regulatory Agency target product profile's minimum acceptable specificity of 95%. Confidence intervals for both tests include the minimum acceptable sensitivity of 80%. In symptomatic patients, negative results on these two POCTs do not preclude the possibility of infection. Tests should not be expected to reliably detect disease more than a week after symptom onset, when viral load may be reduced. ISRCTN142269.


The interface with naturopathy in rural primary health care: a survey of referral practices of general practitioners in rural and regional New South Wales, Australia.
Naturopathy forms an increasingly significant part of the Australian healthcare setting, with high utilisation of naturopaths by the Australian public and a large therapeutic footprint in rural and regional Australia. However, despite these circumstances, there has been little exploration of the interface between naturopathy providers and conventional primary health care practitioners in rural and regional Australia. The study reported here examined the referral practices and factors that underlie referral to naturopaths amongst a sample of rural and regional Australian general practitioners (GPs). A 27-item questionnaire was sent to all 1,486 GPs currently practising in rural and regional Divisions of General Practice in New South Wales, Australia. A total of 585 GPs responded to the questionnaire, with 49 questionnaires returned as "no longer at this address" (response rate: 40.7%). One-quarter of GPs (25.8%) referred to a naturopath at least a few times per year while nearly half (48.8%) stated that they would not refer to a naturopath under any circumstances. GPs were more likely to refer to a naturopath if the GP: was not initially from a rural area (OR=1.78; 95% CI: 0.95, 3.33); believes in the efficacy of naturopathy (OR=5.62; 95% CI: 2.42, 11.36); has seen positive results from naturopathy previously (OR=2.61; 95% CI: 1.35, 5.05); perceives a lack of other treatment options for their patient (OR=5.25; 95% CI: 2.42; 11.36); uses peer-reviewed literature as their major source of CAM information (OR=3.03; 95% CI: 1.65, 5.55); uses CAM practitioners as a major source of CAM information (OR=6.09; 95% CI: 2.91, 12.72); and does not have an existing relationship with any CAM provider (OR=3.03; 95% CI: 1.53, 6.25). There is little interaction (both via referrals as well as the development of professional relationships) between the naturopathic and GP communities in rural and regional Australia, with significant levels of both support and opposition for naturopathic referral amongst GPs. The significant presence and high utilisation of naturopathy in rural primary health care, which appears to work in parallel to conventional medical care rather than in conjunction with it, should serve as an impetus for increased research into naturopathy practice, policy and regulation in rural and regional health.


Relationship between dysfunctional breathing patterns and ability to achieve target heart rate variability with features of "coherence" during biofeedback.
Heart rate variability (HRV) biofeedback is a self-regulation strategy used to improve conditions including asthma, stress, hypertension, and chronic obstructive pulmonary disease. Respiratory muscle function affects hemodynamic influences on respiratory sinus arrhythmia (RSA), and HRV and HRV-biofeedback protocols often include slow abdominal breathing to achieve physiologically optimal patterns of HRV with power spectral distribution concentrated around the 0.1-Hz frequency and large amplitude. It is likely that optimal balanced breathing patterns and ability to entrain heart rhythms to breathing reflect physiological efficiency and resilience and that individuals with dysfunctional breathing patterns may have difficulty voluntarily modulating HRV and RSA. The relationship between breathing movement patterns and HRV, however, has not been investigated. This study examines how individuals' habitual breathing patterns correspond with their ability to optimize HRV and RSA. Breathing pattern was assessed using the Manual Assessment of Respiratory Motion (MARM) and the Hi Lo manual palpation techniques in 83 people with possible dysfunctional breathing before they attempted HRV biofeedback. Mean respiratory rate was also assessed. Subsequently, participants applied a brief 5-minute biofeedback protocol, involving breathing and positive emotional focus, to achieve HRV patterns proposed to reflect physiological "coherence" and entrainment of heart rhythm oscillations to other oscillating body systems. Thoracic-dominant breathing was associated with decreased coherence of HRV (r = -.463, P = .0001). Individuals with paradoxical breathing had the lowest HRV coherence (t(8) = 10.7, P = .001), and the negative relationship between coherence of HRV and extent of thoracic breathing was strongest in this group (r = -.768, P = .03). Dysfunctional breathing patterns are associated with decreased ability to achieve HRV patterns that reflect cardiorespiratory efficiency and autonomic nervous system balance. This suggests that dysfunctional breathing patterns are not only biomechanically inefficient but also reflect decreased physiological resilience. Breathing assessment using simple manual techniques such as the MARM and Hi Lo may be useful in HRV biofeedback to identify if poor responders require more emphasis on correction of dysfunctional breathing.


The impact of routine assessment of past or current mental health on help-seeking in the perinatal period.
Clinical practice guidelines now recommend that women be asked about their past or current mental health as a routine component of maternity care. However, the value of this line of enquiry in increasing engagement with support services, as required, remains controversial. The current study aimed to examine whether assessment of past or current mental health, received with or without referral for additional support, is associated with help-seeking during pregnancy and the postpartum. A subsample of women drawn from the Australian Longitudinal Study on Women's Health (young cohort) who reported experiencing significant emotional distress during pregnancy (N=398) or in the 12 months following birth (N=380) participated in the study. Multivariate analysis showed that women who were not asked about their emotional health were less likely to seek any formal help during both pregnancy (adjOR=0.09, 95%CI: 0.04-0.24) and the postpartum (adjOR=0.07, 95%CI: 0.02-0.13), as were women who were asked about these issues but who were not referred for additional support (antenatal: adjOR=0.26, 95%CI: 0.15-0.45; postnatal: adjOR=0.14, 95%CI: 0.07-0.27). However, considerable levels of consultation with general practitioners, midwives and child health nurses, even in the absence of referral, were evident. This study demonstrates that enquiry by a health professional about women's past or current mental health is associated with help-seeking throughout the perinatal period. The clinical and resource implications of these findings for the primary health care sector should be considered prior to the implementation of future routine perinatal depression screening or psychosocial assessment programmes.


Mega base map of the epidermal growth factor (EGF) receptor gene flanking regions and structure of the amplification units in EGF receptor-hyperproducing squamous carcinoma cells.
We have established a mega base scale physical map of the 5'- and 3'-flanking regions of the epidermal growth factor (EGF) receptor gene using CpG-recognition rare-cutting restriction enzymes and pulsed-field gel electrophoresis. In this map, a methylation-free CpG island (HTF island) is located within an 8-kilobase pair (kb) EcoRI fragment which includes exon 1 of the EGF receptor gene. From this HTF island, a 390-kb NotI fragment was identified as the longest 5'-flanking region and a 540-kb MluI fragment as the longest 3'-flanking region. Utilizing this map information, we have analyzed the structure of the flanking regions of amplified EGF receptor genes which are found in various squamous carcinoma cells. Among seven cell lines tested, four cell lines carrying EGF receptor genes in amounts more than 20 times that of normal cells showed amplification together with large 5'- and 3'-flanking regions. The amplified 5'-flanking regions were rearranged in different forms but were distinct in each cell line. The amplified 3'-flanking regions were at least 540 kb in size and common to all the cell lines, except that A431 had rearrangement points within 540 kb downstream of the HTF island. Thus, the size of amplification units appears to be large and different in each cell line.


Analysis of the resonant frequency of the octagonal split resonant rings with metal wires.
The resonant frequency determined by octagonal split resonant rings (SRRs) is presented and analyzed. The simulated results show that the resonant frequency improves when the opening size of the SRRs becomes larger, and that enhancement of the coupling degree can improve the resonant frequency and widen the band. Four samples are designed to implement the experimental measurement. The results show that the bandwidth of the resonant frequency becomes wider when the thickness of the substrate becomes thinner, and the resonant frequency is shifted by changing the cell size. It is demonstrated that the experimental results show good agreement with the theoretical analysis.


Impact of Cancer and Cardiovascular Disease on In-Hospital Outcomes of COVID-19 Patients: Results From the American Heart Association COVID-19 Cardiovascular Disease Registry.
Background: While pre-existing cardiovascular disease (CVD) appears to be associated with poor outcomes in patients with Coronavirus Disease 2019 (COVID-19), data on patients with CVD and concomitant cancer is limited. Evaluate the effect of underlying CVD and CVD risk factors with cancer history on in-hospital mortality in those with COVID-19. Methods: Data from symptomatic adults hospitalized with COVID-19 at 86 hospitals in the US enrolled in the American Heart Association's COVID-19 CVD Registry was analyzed. The primary exposure was cancer history. The primary outcome was in-hospital death. Multivariable logistic regression models were adjusted for demographics, CVD risk factors, and CVD. Interaction between history of cancer with concomitant CVD and CVD risk factors were tested. Results: Among 8222 patients, 892 (10.8%) had a history of cancer and 1501 (18.3%) died. Cancer history had significant interaction with CVD risk factors of age, body mass index (BMI), and smoking history, but not underlying CVD itself. History of cancer was significantly associated with increased in-hospital death (among average age and BMI patients, adjusted odds ratio [aOR]=3.60, 95% confidence interval [CI]: 2.07-6.24; p<0.0001 in those with a smoking history and aOR=1.33, 95%CI: 1.01 - 1.76; p=0.04 in non-smokers). Among the cancer subgroup, prior use of chemotherapy within 2 weeks of admission was associated with in-hospital death (aOR=1.72, 95%CI: 1.05-2.80; p=0.03). Underlying CVD demonstrated a numerical but statistically nonsignificant trend toward increased mortality (aOR=1.18, 95% CI: 0.99 - 1.41; p=0.07). Conclusion: Among hospitalized COVID-19 patients, cancer history was a predictor of in-hospital mortality. Notably, among cancer patients, recent use of chemotherapy, but not underlying CVD itself, was associated with worse survival. These findings have important implications in cancer therapy considerations and vaccine distribution in cancer patients with and without underlying CVD and CVD risk factors.


Is the combination of 5-fluorouracil and folinic acid effective in advanced gastric carcinoma? Results of a pilot study and review of the literature.
The main purpose of this trial was to investigate the activity of a less toxic regimen in patients with advanced gastric carcinoma regarding response rate and survival. We report on a pilot study of 20 patients treated with the combination of 5-fluorouracil and folinic acid. 5-fluorouracil was administered at a daily dose of 450 mg/m2 followed by folinic acid 200 mg/m2 for four consecutive days every four weeks. The overall response rate was 15% with one complete and two partial responders. The median duration of response was 11, 10 and 10 months respectively. The median survival of all patients was 9 months. Toxicity was mild and consisted primarily of diarrhea. Stomatitis and myelosuppression occurred rarely. In conclusion, the combination of 5-fluorouracil and folinic acid has moderate activity in gastric cancer, while median survival appears similar to the other more intensive regimens. The combination is very well tolerated with minimal toxicity. A detailed review of the literature regarding the effectiveness of the above regimens on response rate, overall survival and toxicity is also presented.


An Interprofessional Collaborative Practice model for preparation of clinical educators.
Work-integrated learning is essential to health professional education, but faces increasing academic and industry resource pressures. The aim of this pilot "Professional Practice Project" was to develop and implement an innovative education intervention for clinical educators across several health disciplines. The project used interprofessional collaboration as its underlying philosophy, and a participatory action research methodology in four cycles: Cycle 1: Formation of an interprofessional project executive and working party from academic staff. Data collection of student insights into work integrated learning. Cycle 2: Formation of an interprofessional reference group to inform curriculum development for a series of clinical education workshops. Cycle 3: Delivery of workshops; 174 clinical educators, supervisors and preceptors attended two workshops: "Introduction to experiential learning" and " utilizing available resources for learning". Cycle 4: Seminar discussion of the Professional Practice Project at a national health-education conference. This pilot project demonstrated the advantages of using collaborative synergies to allow innovation around clinical education, free from the constraints of traditional discipline-specific education models. The planning, delivery and evaluation of clinical education workshops describe the benefits of interprofessional collaboration through enhanced creative thinking, sharing of clinical education models and a broadening of experience for both learners and facilitators.


A global quality assurance system for personalized radiation therapy treatment planning for the prostate (or other sites).
The quality of radiotherapy treatment plans varies across institutions and depends on the experience of the planner. For the purpose of intra- and inter-institutional homogenization of treatment plan quality, we present an algorithm that learns the organs-at-risk (OARs) sparing patterns from a database of high quality plans. Thereafter, the algorithm predicts the dose that similar organs will receive in future radiotherapy plans prior to treatment planning on the basis of the anatomies of the organs. The predicted dose provides the basis for the individualized specification of planning objectives, and for the objective assessment of the quality of radiotherapy plans. One hundred and twenty eight (128) Volumetric Modulated Arc Therapy (VMAT) plans were selected from a database of prostate cancer plans. The plans were divided into two groups, namely a training set that is made up of 95 plans and a validation set that consists of 33 plans. A multivariate analysis technique was used to determine the relationships between the positions of voxels and their dose. This information was used to predict the likely sparing of the OARs of the plans of the validation set. The predicted doses were visually and quantitatively compared to the reference data using dose volume histograms, the 3D dose distribution, and a novel evaluation metric that is based on the dose different test. A voxel of the bladder on the average receives a higher dose than a voxel of the rectum in optimized radiotherapy plans for the treatment of prostate cancer in our institution if both voxels are at the same distance to the PTV. Based on our evaluation metric, the predicted and reference dose to the bladder agree to within 5% of the prescribed dose to the PTV in 18 out of 33 cases, while the predicted and reference doses to the rectum agree to within 5% in 28 out of the 33 plans of the validation set. We have described a method to predict the likely dose that OARs will receive before treatment planning. This prospective knowledge could be used to implement a global quality assurance system for personalized radiation therapy treatment planning.


Clinical Survival of Indirect, Anterior Surface-Retained Fiber-Reinforced Composite Fixed Dental Prosthesis: Up to 3-Year Follow-up.
This study evaluated the performance of indirect, anterior, surface-retained, fiber-reinforced-composite restorations (ISFRCR). Between September-2011 and September-2012, 17 patients (13 females, 4 males, mean age: 40.5) received 17 indirect ISFRCRs (maxilla=4; mandible=13). All restorations were made on a plaster model using unidirectional E-glass fibers (Interlig, Angelus) in combination with a veneering resin composite (Gradia, GC) and cemented adhesively (Choice 2, Bisco). No cavity preparations were made on the abutmentteeth. Before cementation, enamel surfaces were cleaned with pumice and etched with 38% H3PO4 for 30 s, rinsed 30 s. Then, adhesive resin was applied accordingly. Patients were followed every 3 months until May 2014. The evaluation protocol involved technical (chipping, debonding or fracture of tooth/restoration) and biological failures (caries). Survival (Kaplan Meier) and annual failure rates were calculated. Mean observation time was 34.6 months. Altogether, 5 failures were observed [survival rate: 70.5%] of which 3 were debondings and 2 delaminations of the veneering composite (chipping). All defective restorations were repaired or recemented, except one, which was remade.


Soy isoflavones and risk of cancer recurrence in a cohort of breast cancer survivors: the Life After Cancer Epidemiology study.
Soy isoflavones, structurally similar to endogenous estrogens, may affect breast cancer through both hormonally mediated and non-hormonally related mechanisms. Although the effects of soy are not well understood, some breast cancer survivors increase their soy intake post-diagnosis in attempt to improve their prognosis. Therefore, we examined the role of soy isoflavone intake and the risk of breast cancer recurrence by hormone receptor status, menopausal status, and tamoxifen therapy. A cohort of 1,954 female breast cancer survivors, diagnosed during 1997-2000, was prospectively followed for 6.31 years and 282 breast cancer recurrences were ascertained. Isoflavone intake was assessed by mailing modified Block and supplemental soy food frequency questionnaires to participants, on average 23 months post-diagnosis. Risk of breast cancer recurrence, measured by hazard ratios (HR) and 95% confidence intervals (CI), was estimated using multivariable delayed entry Cox proportional hazards models. Suggestive trends for a reduced risk of cancer recurrence were observed with increasing quintiles of daidzein and glycetin intake compared to no intake among postmenopausal women (P for trend: P = 0.08 for daidzein, P = 0.06 for glycetin) and among tamoxifen users (P = 0.10 for daidzein, P = 0.05 for glycetin). Among postmenopausal women treated with tamoxifen, there was an approximately 60% reduction in breast cancer recurrence comparing the highest to the lowest daidzein intakes (>1,453 vs. <7.7 microg/day; HR, 0.48; 95% CI, 0.21-0.79, P = 0.008). Soy isoflavones consumed at levels comparable to those in Asian populations may reduce the risk of cancer recurrence in women receiving tamoxifen therapy and moreover, appears not to interfere with tamoxifen efficacy. Further confirmation is required in other large prospective studies before recommendations regarding soy intake can be issued to breast cancer survivors.


Is GDF5 gene promoter polymorphism +104T/C associated with osteoarthritis in the Eastern of Turkey population?
Osteoarthritis (OA) is the most common form of arthritis. Genetic factors have been shown to play important roles in the etiology of OA. The gene growth differentiation factor 5 (GDF5) has been implicated in skeletal development and joint morphogenesis in human and mice. A functional single nucleotide polymorphism (SNP) +104T/C in the 5'-UTR of GDF5 (rs143383) was reported to be associated with osteoarthritis susceptibility in Han Chinese and Japanese populations. Our objective was to assess whether this SNP was also associated with OA in the Eastern Turkey population.A total of 172 cases including 95 patients with idiopathic OA and 77 control cases were recruited into the study. DNA samples were extracted from peripheral blood lymphocytes of all cases by using salting out method. The +104T/C polymorphism was genotyped by PCR-RFLP method. In terms of genotype comparison there wasn't any correlation between patient and control groups. Frequency of C allele was found to be higher in-patient group than control group and statistical analysis showed a poor correlation in allele frequencies of the +104T/C SNP of GDF5 gene between cases and controls (p<0.05). Significant correlation between GDF5 and OA has been reported in Asian population, especially T alleles were found in higher frequencies and related to OA. Our study did not confirm this association and also in term of T allele. Interestingly, we found higher frequency of C allele in patient group than control group and our results are compatible with the study carried out in Greek population.


Association of serum YKL-40 levels with urinary albumin excretion rate in young Japanese patients with type 1 diabetes mellitus.
YKL-40 is a marker of inflammation and endothelial dysfunction, both of which play important roles in the progression of diabetic complications. However, little information has been obtained about serum YKL-40 levels in type 1 diabetic patients. We evaluated YKL-40 levels and its association with diabetic micro- and macroandgiopathy in 131 young Japanese type 1 diabetic patients without advanced diabetic complications (aged 24.7±5.9 years) and 97 age- and gender-matched healthy controls. YKL-40 levels were significantly elevated in type 1 diabetic patients than in healthy controls (median (range) 46.4 (20.3-136.7) and 52.3 (21.4-274.1) ng/mL, respectively, p = 0.003). There was a significant positive association between YKL-40 levels and urinary albumin creatinine ratio (UACR) (r = 0.226, p = 0.013). Furthermore, a multivariate regression analysis demonstrated that YKL-40 levels were a determinant of UACR independently of conventional risk factors. In addition, YKL-40 levels were significantly higher in participants with diabetic retinopathy compared to those without it (median (range) 55.5 (23.3-274.1) and 50.3 (21.4-237.4) ng/mL, respectively, p = 0.039). Serum YKL-40 levels were elevated in type 1 diabetic patients and associated with increasing level of albuminuria. YKL-40 could be a predictor to assess the risk of diabetic microangiopathy in the early stage in type 1 diabetic patients.


Combination of Two Targeted Medications (Bevacizumab Plus Cetuximab) Improve the Therapeutic Response of Pancreatic Carcinoma.
The objective of this study is to evaluate the efficacy and safety profiles of the targeted medications, bevacizumab and cetuximab, in combination with cytostatic drugs in patients with locally advanced or metastatic pancreatic cancer. In this retrospective phase 2 study, a total of 59 patients with pancreatic cancer were recruited and received conventional (gemcitabine, cisplatin, and fluorouracil) or targeted regimen (conventional plus bevacizumab and cetuximab for the first cycle) in 2-week intervals for four cycles. The primary end-point for this study was the overall response rate. Secondary end-points were progression-free survival and the safety profiles of the combined therapy. The median time-to-progression and overall survival were 3 and 7 months, respectively, in the conventional treatment group as well as 11 and 13 months, respectively, in the targeted medications treatment group. The most common adverse events in both treatment groups were nausea and vomiting. Moderate (Grade 2) nausea and vomiting were more common in the conventional group than the targeted group but severe (Grade 3) nausea and vomiting were more common in the targeted group. Bevacizumab and cetuximab in combination with gemcitabine, cisplatin, and fluorouracil may help lengthen overall survival up to six months for patients with pancreatic cancer.


Emergency Department (ED), ED Observation, Day Hospital, and Hospital Admissions for Adults with Sickle Cell Disease.
Use of alternative venues to manage uncomplicated vaso-occlusive crisis (VOC), such as a day hospital (DH) or ED observation unit, for patients with sickle cell anemia, may significantly reduce admission rates, which may subsequently reduce 30-day readmission rates. In the context of a two-institution quality improvement project to implement best practices for management of patients with sickle cell disease (SCD) VOC, we prospectively compared acute care encounters for utilization of 1) emergency department (ED); 2) ED observation unit; 3) DH, and 4) hospital admission, of two different patient cohorts with SCD presenting to our two study sites. Using a representative sample of patients from each institution, we also tabulated SCD patient visits or admissions to outside hospitals within 20 miles of the patients' home institutions. Over 30 months 427 patients (297 at Site 1 and 130 at Site 2) initiated 4,740 institutional visits, totaling 6,627 different acute care encounters, including combinations of encounters. The range of encounters varied from a low of 0 (203 of 500 patients [40.6%] at Site 1; 65 of 195 patients [33.3%] at Site 2), and a high of 152 (5/month) acute care encounters for one patient at Site 2. Patients at Site 2 were more likely to be admitted to the hospital during the study period (88.4% vs. 74.4%, p=0.0011) and have an ED visit (96.9% vs. 85.5%, p=0.0002). DH was used more frequently at Site 1 (1.207 encounters for 297 patients at Site 1, vs. 199 encounters for 130 patients at Site 2), and ED observation was used at Site 1 only. Thirty-five percent of patients visited hospitals outside their home academic center. In this 30-month assessment of two sickle cell cohorts, healthcare utilization varied dramatically between individual patients. One cohort had more hospital admissions and ED encounters, while the other cohort had more day hospital encounters and used a sickle cell disease observation VOC protocol. One-third of patients sampled visited hospitals for acute care outside of their care providers' institutions.


Intensity and polarization of the radiation emerging from a plane-parallel atmosphere containing monodispersed aerosols.
A method for evaluating the intensity, degree of polarization, direction of polarization, and ellipticity of the scattered radiation emerging from a plane-parallel atmosphere containing large spherical particles is described. In this method, all the elements of the normalized phase matrix are represented by fourier series whose maximum required number of terms depend upon the zenith angles of the directions of the incident and scattered radiation. Some results are presented for an atmospheric model containing water spheres with size parameter 10.0 to show that this method can be used to evaluate reliably all the characteristics of the emergent radiation in a reasonable amount of computer time.


True posterior communicating artery aneurysms: are they more prone to rupture? A biomorphometric analysis.
Posterior communicating artery (PCoA) aneurysms can occur at the junction with the internal carotid artery, posterior cerebral artery (PCA), or the proximal PCoA itself. Hemodynamic stressors contribute to aneurysm formation and may be associated with parent vessel size and aneurysm location. This study evaluates the correlation of various biomorphometric characteristics in 2 of the aforementioned types of PCoA aneurysms. Patients with PCoA aneurysms were analyzed using CT angiography. Source images and reconstructions were used to determine which aneurysms originated purely from the PCoA and those that originated from the internal carotid artery/PCoA junction. Morphometric analysis was performed on the aneurysm, the precommunicating segment of the PCA (P(1)), the ambient segment of the PCA (P(2)), and both PCoA arteries and were correlated to clinical presentation. Parametric and nonparametric analyses were performed to test for significance. A total of 77 PCoA aneurysms were analyzed, and 10 were found to be true PCoA aneurysms (13.0%). The ipsilateral PCoA/P(1) ratio (1.77 +/- 0.44 vs 0.82 +/- 0.46, p = 0.0001) and ipsilateral P(2)/P(1) ratio (1.73 +/- 0.40 vs 1.22 +/- 0.41, p = 0.0003) were significantly larger in true PCoA aneurysms. Interestingly, aneurysm size was statistically larger in the junctional aneurysms (0.14 +/- 0.1 vs 0.072 +/- 0.04 cm(3), p = 0.03). The prevalence of ruptured aneurysms was similar in both groups (approximately 80%, p value not significant). These data suggest that true PCoA aneurysms have a larger PCoA relative to the ipsilateral P(1) segment. To the authors' knowledge, this represents the first such biomorphometric comparison of these different types of PCoA aneurysms. Although statistically smaller in size, true PCoA aneurysms also have a similar prevalence of presenting as a ruptured aneurysm, suggesting that they might be more prone to rupture than a junctional aneurysms of similar size. Further analysis will be required to determine the biophysical factors affecting rupture rates.


Independent component analysis for biomedical signals.
Independent component analysis (ICA) is increasing in popularity in the field of biomedical signal processing. It is generally used when it is required to separate measured multi-channel biomedical signals into their constituent underlying components. The use of ICA has been facilitated in part by the free availability of toolboxes that implement popular flavours of the techniques. Fundamentally ICA in biomedicine involves the extraction and separation of statistically independent sources underlying multiple measurements of biomedical signals. Technical advances in algorithmic developments implementing ICA are reviewed along with new directions in the field. These advances are specifically summarized with applications to biomedical signals in mind. The basic assumptions that are made when applying ICA are discussed, along with their implications when applied particularly to biomedical signals. ICA as a specific embodiment of blind source separation (BSS) is also discussed, and as a consequence the criterion used for establishing independence between sources is reviewed and this leads to the introduction of ICA/BSS techniques based on time, frequency and joint time-frequency decomposition of the data. Finally, advanced implementations of ICA are illustrated as applied to neurophysiologic signals in the form of electro-magnetic brain signals data.


The marine herring myxozoan Ceratomyxa auerbachi (Myxozoa: Ceratomyxidae) uses Chone infundibuliformis (Annelida: Polychaeta: Sabellidae) as invertebrate host.
Sequencing of SSU rDNA showed that actinospores of the tetractinomyxon type, which develop in Chone infundibuliformis Krøyer (Annelida, Polychaeta, Sabellidae) from the northern Øresund, Denmark, are identical with Ceratomyxa auerbachi Kabata, 1962 (Myxozoa, Ceratomyxidae). This myxosporean was found in the gallbladder of the Atlantic herring Clupea harengus L. from the northern Øresund, Denmark, and from the Bergen area, western Norway. The pansporocysts and actinospores of C. auerbachi are described. This is the third elucidated two-host life cycle of a marine myxozoan, and the first involving a marine ceratomyxid.


Evidence for the circulation and inter-hemispheric movement of the H14 subtype influenza A virus.
Three H14 influenza A virus (IAV) isolates recovered in 2010 during routine virus surveillance along the Mississippi Migratory Bird Flyway in Wisconsin, U.S.A. raised questions about the natural history of these rare viruses. These were the first H14 IAV isolates recovered in the Western Hemisphere and the only H14 IAV isolates recovered since the original four isolates in 1982 in Asia. Full length genomic sequencing of the 2010 H14 isolates demonstrated the hemagglutinin (HA) gene from the 1982 and 2010 H14 isolates showed 89.6% nucleotide and 95.6% amino acid similarity and phylogenetic analysis of these viruses placed them with strong support within the H14 subtype lineage. The level of genomic divergence observed between the 1982 and 2010 viruses provides evidence that the H14 HA segment was circulating undetected in hosts and was not maintained in environmental stasis. Further, the evolutionary relationship observed between 1982 H14 and the closely related H4 subtype HA segments were similar to contemporary comparisons suggesting limited adaptive divergence between these sister subtypes. The nonstructural (NS) segment of one 2010 isolate was placed in a NS clade isolated infrequently over the last several decades that includes the NS segment from a previously reported 1982 H14 isolate indicating the existence of an unidentified pool of genomic diversity. An additional neuraminidase reassortment event indicated a recent inter-hemispheric gene flow from Asia into the center of North America. These results demonstrate temporal and spatial gaps in the understanding of IAV natural history. Additionally, the reassortment history of these viruses raises concern for the inter-continental spread of IAVs and the efficacy of current IAV surveillance efforts in detecting genomic diversity of viruses circulating in wild birds.


The effect of vanadate on human kidney potassium dependent phosphatase.
This study examined the effects of vanadate on the potassium dependent phosphatase activity present in purified human kidney microsomal (Na+ + K+)-adenosine triphosphatase. Vanadate anion inhibited the K+-dependent phosphatase at a K1 of 35 nM. This inhibition was noncompetitive with the substrate, p-nitrophenylphosphate. The inhibition by vanadate at 1 mM K+ was only 45% of the inhibition that was observed at 10 mM K+. Neither preincubation of the enzyme with vanadate, nor changing the pH of the assay from 8.2 to 7.2 had any effect on the K1 for vanadate. The inclusion of 2.5 mM isoproterenol, to complex the yanadate, reversed the inhibition, as did diluting the enzymatic reaction. Vanadate also inhibited the overall (Na+ + K+)-ATPase reaction at a K1 of 1.91 microM. This inhibition was also reversible upon inclusion of isoproterenol in the assay. Increasing the level of magnesium from 6 mM to 30 mM lowered the K1 of vanadate to 0.25 microM. The possible role of vanadate as a physiological mediator of (Na+ + k+)-atpase activity is discussed.


Protein enhancement of hammerhead ribozyme catalysis.
When the recognition sequence of a ribozyme is extended beyond a certain length, turnover is slowed and specificity is decreased. Here, it is shown that a protein can help a ribozyme overcome these general limitations on ribozyme activity. Cleavage of an RNA oligonucleotide by a hammerhead ribozyme is enhanced 10- to 20-fold upon addition of a protein derived from the p7 nucleocapsid (NC) protein of human immunodeficiency virus-type 1. The NC protein also enhances the ability of the ribozyme to discriminate between cleavage of RNA oligonucleotides with differing sequences. These catalytic improvements can be attributed to the strand exchange activity of this RNA binding protein. It is conceivable that endogenous or added proteins may provide analogous increases in ribozyme activity and specificity in vivo.


Oridonin induces apoptosis in human oral cancer cells via phosphorylation of histone H2AX.
Oridonin, a natural diterpenoid purified from Rabdosia rubescens, has displayed beneficial biological activities, including anti-proliferation and anti-angiogenesis effects, in various types of cancers. However, the anti-cancer potential of oridonin and its mechanism in oral cancer have never previously been studied. In this study, we assessed the role of oridonin as an inducer of apoptosis in HSC-3 and HSC-4 human oral cancer cells. Our results showed that oridonin reduces the viability of human oral cancer cells and significantly increases the expression of γH2AX, a well-known marker of DNA damage. 4',6-Diamidino-2-phenylindole (DAPI) staining and western blotting showed that oridonin causes nuclear condensation and fragmentation, and induces cleavage of poly(ADP-ribose) polymerase (PARP). Moreover, oridonin-induced γH2AX accumulation was partially abrogated by Z-VAD, a pan-caspase inhibitor. Taken together, our results suggest that oridonin can effectively induce apoptosis by augmenting the expression of γH2AX in response to DNA damage and might be a promising anti-cancer drug candidate for the treatment of oral cancer.


Pathway of care among psychiatric patients attending a mental health institution in central India.
Only a limited proportion of patients with psychiatric disorders attend the healthcare facilities, and that too when the condition becomes severe. Treatment from unqualified medical practitioners and faith healers is a common practice, and is attributable to the delay in proper treatment. A cross-sectional study was conducted to understand the pathway of care adopted by psychiatric patients and its relationship with the socio-demographic determinants in the study population. The subjects were selected from urban specialty psychiatric hospitals and interviewed using a pre-tested, semi-structured interview schedule. The data was analyzed using SPSS v10.0 software. The Chi square test, T test, and Kruskall Wallis Test were used, as needed. A total of 295 patients (203 males) were included in this study. The majority of the patients (45%) were suffering from Bipolar affective disorders (45%), followed by schizophrenia (36%). The majority, 203 (68%), were from the rural area, with 94 patients being illiterate. The mean distance traveled for treatment was 249 km. The majority of these (69%) had first contacted faith healers and a qualified psychiatrist was the first contacted person for only 9.2% of the patients. A large proportion of psychiatric patients do not attend any health facility due to a lack of awareness about treatment services, the distance, and due to the fear of the stigma associated with treatment. The psychiatric patients first seek the help of various sources prior to attending a psychiatric health facility. The pathway adopted by these patients need to be kept in mind at the time of preparation of the mental health program.


Factors associated with sexual abstinence among adolescents in four sub-Saharan African countries.
Drawing on nationally representative data collected from Burkinabé, Ghanaian, Malawian, and Ugandan adolescents, this study examines differences among four groups of never married, 15-19 year olds: primary abstainers (sexually inexperienced), secondary abstainers (last sex more than 12 months prior to the survey), recent abstainers (sexually active in last year but not in the last 3 months), and sexually active (had sexual intercourse in the last 3 months). The percentage of primary abstinent adolescents ranged from 42% (Malawian males) to 85% (Ghanaian males). In general, a greater proportion of females than males were primary abstainers. Primary abstainers were younger than sexually experienced adolescents. Current involvement in a romantic relationship was a significant predictor of sexual status with primary abstainers being the least likely to be romantically involved. Overall, findings suggest that adolescents' gender, prior sexual experiences and contextual circumstances, such as romantic partnerships, should be considered when designing abstinence promotion programs.


Tunable antimicrobial polypropylene surfaces: simultaneous attachment of penicillin (Gram +) and gentamicin (Gram -).
Surface reactions were performed on polypropylene (PP) surfaces to retard the simultaneous growth of Staphylococcus aureus (S. aureus) and Pseudomonas putida (P. putida) bacteria. Microwave plasma reactions in the presence of maleic anhydride (MA) resulted in the formation of acid groups on the surface of PP. Such surfaces were further modified by conducting two parallel reactions: (1) poly(ethylene glycol) (PEG) was attached to COOH groups of the PP surface, followed by penicillin V (PEN) reactions to target S. aureus destruction and (2) diglycidyl PEG was attached, followed by gentamicin (GEN) reactions, to create antimicrobial surfaces targeted at P. putida . Simultaneous gram "+" and gram "-" resistance was obtained by varying the PEN/GEN ratios on such modified PP surfaces, thus providing the controllable degree of gram "+" and gram "-" antimicrobial strength. While spectroscopic analyses revealed chemical attachments of PEN and GEN, the effectiveness against proliferation of S. aureus (Gram +) and P. putida (Gram -) bacteria was determined using liquid culture tests. These studies show for the first time the formation of tunable antimicrobial polypropylene surfaces with controllable strength.


Large-scale influenza vaccination promotion on a mobile app platform: A randomized controlled trial.
While health-care providers have used incentives in an attempt to motivate patients to obtain vaccinations, their effect on vaccination rates has not been systematically evaluated on a large scale. In this study, we examined whether mobile applications may improve population vaccination rates through enhanced communication and incentives education. Our study is the first randomized controlled trial assessing the effect of large-scale messaging combined with individualized incentives on influenza-vaccination rates. In this trial, we delivered messages regarding influenza vaccinations to 50,286 adults, aged 18 through 65, then compared the subsequent vaccination rate, the effectiveness of the message content and the timing. Multiple rounds of messaging occurred over a seven-week period during the 2016 flu season, after which vaccination rates were observed for one week. Participants were randomly assigned to one of three messaging approaches: conspicuous (highlighting the amount of rewards to be received for obtaining a flu shot); generic (promoting vaccinations with no mention of rewards); or no-message. Evidence of vaccination obtainment was indicated by medical and pharmacy claims, augmented by patients self-reporting through the mobile wellness app during the study period. Of the people assigned to receive messaging, 23.2% obtained influenza vaccination, compared to 22.0% of people who obtained vaccination in the no-messaging control arm. This difference was statistically significant (p < 0.01). The research revealed that messaging effectiveness decreased after each successive batch sent, suggesting that most participants responsive to messaging would become activated immediately after receiving one alert. Interestingly, in this large-scale study, there were no significant differences between conspicuous incentives and generic messaging, suggesting an important area for future research. Trial Registration: clinicaltrials.gov identifier: NCT02908893.


Genetic investigation of methylenetetrahydrofolate reductase (MTHFR) and catechol-O-methyl transferase (COMT) in multiple sclerosis.
Multiple sclerosis (MS) is a chronic neurological disease characterized by central nervous system (CNS) inflammation and demyelination. The C677T substitution variant in the methylenetetrahydrofolate reductase (MTHFR) gene has been associated with increased levels of circulating homocysteine and is a mild risk factor for vascular disease. Higher blood levels of homocysteine have also been reported in MS. Thus, the C677T mutation of the MTHFR gene may influence MS susceptibility. Noradrenaline, a neurotransmitter believed to play an immunosupressive role in neuroinflammatory disorders, is catabolized by catechol-O-methyl transferase (COMT). The COMT G158A substitution results in a three- to four-fold decreased activity of the COMT enzyme, which may influence CNS synaptic catecholamine breakdown and could also play a role in MS inflammation. We tested DNA from Australian MS patients and unaffected control subjects, matched for gender, age and ethnicity. Specifically, we genotyped the MTHFR C677T and the COMT G158A mutations. Genotype distributions showed that the homozygous mutant MTHFR genotype (T/T) and the COMT (H/H) genotype were slightly over-represented in the MS group (16% versus 11% and 24% versus 19%, respectively), but both variations failed to reach statistical significance (P=0.15 and P=0.32, respectively). Hence, results from the present study do not support a major role for either functional gene mutation in MS susceptibility.


A spectacular new Philippine monitor lizard reveals a hidden biogeographic boundary and a novel flagship species for conservation.
As humans continue to explore the last uncharted regions of the planet, discoveries of previously unknown species of large vertebrates have become infrequent. Here, we report on the discovery of a spectacular new species of giant, secretive, frugivorous, forest monitor lizard (Genus: Varanus) from the forests of the northern Philippines. Using data from morphology and mitochondrial and nuclear DNA sequences, we demonstrate the taxonomic distinctiveness of this new 2 m long species and provide insight into its historical biogeography and systematic affinities. Our molecular phylogenetic analyses indicate that the new species is closely related to Varanus olivaceus (from southern Luzon and nearby islands), but it differs from this and other varanids with respect to characteristics of scalation, colour pattern, body size, anatomy of the reproductive organs and genetic divergence. The new species appears to be restricted to forests of the central and northern Sierra Madre mountain range; it is separated from the range of V. olivaceus by a more than 150 km stretch that includes at least three low-elevation river valley barriers to dispersal. This discovery identifies a seldom-perceived biogeographic boundary and emphasizes the need for continued biodiversity research in the megadiverse conservation hotspot of the Philippines. It is anticipated that the new species will serve as an important flagship species for conservation efforts aimed at preserving the remaining forests of northern Luzon.


Long-term Outcomes of Laparoscopic Versus Open Surgery for Clinical Stage I Gastric Cancer: The LOC-1 Study.
Clinical trials comparing laparoscopic gastrectomy (LG) versus traditional open gastrectomy (OG) have been planned, their surgical outcomes reported but their oncologic outcomes are still pending. Consequently, we have conducted this large-scale historical cohort study to provide relevant information rapidly to guide our current practice. Through a consensus meeting involving surgeons, biostatisticians, and epidemiologists, 30 variables of preoperative information possibly influencing surgeons' choice between LG versus OG and potentially associating with outcomes were identified to enable rigorous estimation of propensity scores. A total of 4235 consecutive patients who underwent gastrectomy for gastric adenocarcinoma were identified and their relevant data were gathered from the participating hospitals. After propensity score matching, 1848 patients (924 each for LG and OG) were selected for comparison of long-term outcomes. In the propensity-matched population, the 5-year overall survival was 96.3% [95% confidence interval (CI) 95.0-97.6] in the OG as compared with 97.1% (95% CI, 95.9-98.3) in LG. The number of all-cause death was 33/924 in the OG and 24/924 in the LG through the entire period, and the hazard ratio (LG/OG) for overall death was 0.75 (95% CI, 0.44-1.27; P = 0.290). The 3-year recurrence-free survival was 97.4% (95% CI, 96.4-98.5) in the OG and 97.7% (95% CI, 96.5-98.8) in the LG. The number of recurrence was 22/924 in the OG and 21/924 in the LG through the entire period, and the hazard ratio was 1.01 (95% CI, 0.55-1.84; P = 0.981). This observational study adjusted for all-known confounding factors seems to provide strong enough evidence to suggest that LG is oncologically comparable to OG for gastric cancer.


Molecular basis of length polymorphism in the human zeta-globin gene complex.
The length polymorphism between the human zeta-globin gene and its pseudogene is caused by an allele-specific variation in the copy number of a tandemly repeating 36-base-pair sequence. This sequence is related to a tandemly repeated 14-base-pair sequence in the 5' flanking region of the human insulin gene, which is known to cause length polymorphism, and to a repetitive sequence in intervening sequence (IVS) 1 of the pseudo-zeta-globin gene. Evidence is presented that the latter is also of variable length, probably because of differences in the copy number of the tandem repeat. The homology between the three length polymorphisms may be an indication of the presence of a more widespread group of related sequences in the human genome, which might be useful for generalized linkage studies.


A search for viable organisms in a lunar sample.
The hypothesis that the moon could harbor viable life forms was not verified on analysis of the first samnples from the Apollo 11 mission. Biological examnination of 50 grainis of the butlk fines confirmn the negative results obtained by the Manned Spacecraft Center quarantine teamyz. No viable life forms, including terrestrial contaminants, were found when the sample was tested in 300 separate environmtenits. Only colored illorganiic artifacts, resembling mnicrobial clonies, appeared aroun1cd some particles. Manned Spacecraft Center, Houston.


Safety of uninterrupted anticoagulation in patients requiring elective coronary angiography with or without percutaneous coronary intervention: a systematic review and metaanalysis.
Patients who are receiving vitamin K antagonist (VKA) therapy pose challenges when they require surgery or invasive procedures because the risk for bleeding during the procedure must be balanced against the risk of an atherothrombotic event if the VKA is interrupted. However, it may be possible to safely perform some procedures, such as coronary angiography with or without percutaneous coronary intervention (PCI), without VKA interruption. We undertook a systematic review and metaanalysis to assess the safety of a periprocedural management strategy of uninterrupted VKA (U-VKA) vs interrupted VKA (I-VKA) with or without bridging with low-molecular-weight heparin in patients undergoing elective coronary angiography with or without PCI. Eight studies were included in the review. Most were of moderate to very low quality. A strategy of U-VKA appears to confer approximately one-half the risk (odds ratio, 0.43; 95% CI, 0.26-0.73) of experiencing an access site bleeding complication within 1 week of the procedure compared with a strategy of I-VKA. The U-VKA strategy was associated with a pooled access site bleeding complication rate of 4.0% (95% CI, 3.0-7.0), and although high heterogeneity precluded pooling of such a rate in the I-VKA group, these rates ranged from 2% to 14%. Although it appears that coronary angiography with or without PCI can be safely performed without interrupting VKA, the low methodologic quality of existing studies precludes any definitive conclusions. Randomized trials assessing different anticoagulation strategies are needed to establish evidence-based practice guidelines in this setting.


Perceived symptoms in the recognition of hypoglycemia.
For people with diabetes, detection of hypoglycemic symptoms is a critical tool for the recognition and treatment of hypoglycemia. This is not a simple process involving only the occurrence of a hypoglycemic-relevant physiological event, such as sweating. We propose a four-step biological and psychological model that leads to the accurate recognition of hypoglycemia through symptoms. The model illustrates both the chronic and transient modifiers that can enhance and interfere with recognition of hypoglycemia. Three common methods are used to investigate the occurrence and utility of hypoglycemic symptoms that relate to this model. This article reviews the advantages and disadvantages of these methods, providing previously unpublished illustrative data. The field study hit/false alarm approach was shown to be the most useful method. The relevance of this model and data to the concept of hypoglycemic awareness/unawareness and blood glucose awareness training is discussed.


Blood-brain barrier: penetration of morphine, codeine, heroin, and methadone after carotid injection.
Labeled morphine, codeine, heroin, or methadone was injected as a bolus into the common carotid artery of the rat, and the rat was decapitated 15 seconds later. The brain uptake of the drug was calculated by measurement of the brain content of the drug as a percentage of a labeled, highly diffusible reference substance simultaneously injected. The uptake of morphine was below measurability; the uptake of codeine was 24 percent; heroin, 68 percent; and methadone, 42 percent. Brain uptakes of morphine and codeine were also studied after intravenous injection and correlated well with uptakes after carotid injection; the uptake of codeine being nearly complete by 30 seconds. These studies indicate that brain uptake of certain of these drugs is very rapid and that uptake of heroin injected intravenously is probably limited by the regional flow of blood in the brain. The possible relation of this rapid penetration of the blood-brain barrier by heroin to its strongly addictive properties is discussed.


Randomized Study of Assessment Effects on Alcohol Use by Persons With HIV in Rural Uganda.
Unhealthy alcohol use is a crucial driver of HIV in sub-Saharan Africa, and interventions are needed. The goal of this study was to assess whether assessment itself (assessment reactivity) causes declines in alcohol use in a research study in persons with HIV in Uganda. Study participants were adult patients of the Immune Suppression Syndrome (ISS) Clinic in Mbarara, Uganda, who were new to HIV care and reported any alcohol consumption in the prior year. Participants were randomized to (a) a study cohort, with structured interviews, breath alcohol analysis tests, and blood draws conducted quarterly, or (b) a minimally assessed arm that engaged in these procedures only once, at 6 months after baseline. The main outcome was unhealthy drinking at 6 months, defined as Alcohol Use Disorders Identification Test-Consumption [AUDIT-C] positive (≥3 for women, ≥4 for men) or phosphatidylethanol (PEth; an alcohol biomarker) level ≥ 50 ng/ml. We also examined this outcome stratified by gender. We examined 175 and 139 persons in the quarterly assessed versus minimally assessed arms, respectively. Overall, 54.8% were male, the median age was 30 (interquartile range: 25-36), and 58.0% initiated anti-retroviral therapy at 6 months. Nearly equal proportions (53.7% and 51.1% in the study quarterly assessed vs. minimally assessed arm, respectively) engaged in unhealthy drinking in the 3 months before the 6-month study visit (p = .64), and we found no evidence of interaction by gender (p = .36). We found no evidence of assessment reactivity in a study that included quarterly study visits. Assessment is not sufficient to act as an intervention itself in this population with high levels of unhealthy drinking. Interventions are needed to decrease alcohol consumption in this population.


Rh-Catalyzed Decarbonylation of Conjugated Ynones via Carbon-Alkyne Bond Activation: Reaction Scope and Mechanistic Exploration via DFT Calculations.
In this full article, detailed development of a catalytic decarbonylation of conjugated monoynones to synthesize disubstituted alkynes is described. The reaction scope and limitation has been thoroughly investigated, and a broad range of functional groups including heterocycles were compatible under the catalytic conditions. Mechanistic exploration via DFT calculations has also been executed. Through the computational study, a proposed catalytic mechanism has been carefully evaluated. These efforts are expected to serve as an important exploratory study for developing catalytic alkyne-transfer reactions via carbon-alkyne bond activation.


A genome-wide association study reveals a quantitative trait locus for age at first calving in delta/notch-like EGF repeat containing on chromosome 2 in Japanese Black cattle.
Age at first calving (AFC) is an important trait for achieving earlier reproductive performance in cattle. To identify quantitative trait loci for AFC in Japanese Black cattle, we conducted a genome-wide association study using 866 animals with extreme AFC values selected from a larger group of 52, 009 animals. We identified single nucleotide polymorphisms (SNPs) on bovine chromosome 2 that were associated with AFC. These SNPs were located within 112.8-kbp intronic region of delta/notch-like EGF repeat containing (DNER) and proved to be in a state of high linkage disequilibrium. The association was replicated in an independent sample of 2963 animals. In the replicated population, the frequency of the reduced AFC allele (Q) was 0.463, and the allele accounts for 8% of the total genetic variance. The effect of allele substitution on AFC was a decrease of 11.54 days. The results suggest that the Q allele could serve as a useful marker in Japanese Black cattle to select animals with superior AFC performance.


The evolutionary history of TLR4 polymorphisms in Europe.
Infections exert important evolutionary pressures shaping the human genome, especially on genes involved in host defense. A crucial step for host defense is recognition of pathogens by pattern recognition receptors on innate immune cells, among which Toll-like receptor 4 (TLR4) is one of the best known. Genetic variation in TLR4 (Asp299Gly, Thr399Ile) has been recently described. Haplotype frequencies of these polymorphisms differ among African, Asian and European populations, suggesting evolutionary pressures exerted by local infections. The TLR4 299Gly/399Ile haplotype, characteristic mainly of European populations, has relatively high frequency in the Iberian peninsula. This region is also described as refuge area during the last glacial maximum 20,000 years ago, from which repopulation of Europe took place. We speculate that a genetic bottleneck in the Iberian peninsula could have promoted the increased frequency of this haplotype by genetic drift. This hypothesis is supported by three arguments: (1) the West-East gradient of prevalence in the haplotype among European populations; (2) ancient DNA from Neolithic burials in the Iberian peninsula, dated 6,600-4,500 years before present, confirmed the relatively high frequency of this haplotype in the region, and (3) no functional differences between this haplotype and wild-type TLR4 have been found. In contrast, the disappearance of the 299Gly/399Thr haplotype in Europe is most likely due to negative selection due to sepsis. In conclusion, differences in distribution of TLR4 polymorphisms Asp299Gly and Thr399Ile in European populations are most likely due to a combination of population migration events combined with selection due to sepsis.


Nonmechanical transverse scanning laser Doppler velocimeter using wavelength change.
A transverse scanning laser Doppler velocimeter (LDV) that does not require any moving mechanism in its sensor probe is proposed, and the scanning function is demonstrated theoretically and experimentally. In the proposed scanning LDV, the measurement position is transversely scanned on the basis of a wavelength change induced by a tunable laser and a combination of a grating and a Dove prism. To demonstrate the scanning function in the transverse direction, an experiment was carried out using a setup of the sensor probe consisting of bulk optical components. The experimental results indicate that a transverse scanning function was successfully obtained. The scanning range in the vertical direction is estimated to be 11.3 mm over wavelengths of 1520 to 1570 nm.


Expression of N-methyl-D-aspartate receptor subunits in the bovine ovum: ova as a potential source of autoantigens causing anti-NMDAR encephalitis.
Autoimmune synaptic encephalitis is characterized by the presence of autoantibodies against synaptic constituent receptors and manifests as neurological and psychiatric disorders. Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is such an autoimmune disorder that predominantly affects young women. It is associated with antibodies against the extracellular region of the NR1 subunit of postsynaptic NMDAR. Each NMDAR functions as a heterotetrameric complex that is composed of four subunits, including NR1 and NR2A, NR2B, or NR2C. Importantly, ovarian teratoma is a typical complication of anti-NMDAR encephalitis in female patients and may contain antigenic neural tissue; however, antigenic sites remain unknown in female patients without ovarian teratoma. The purpose of this study was to investigate the expression of NMDARs in the ovum. We detected NR1 and NR2B immunoreactivity in protein fractions extracted from the bovine ovary and ova by SDS-polyacrylamide gel electrophoresis and immunoblotting analysis. Immunoprecipitates digested with trypsin were analyzed by reverse phase liquid chromatography coupled to tandem mass spectrometry. We obtained the following five peptides: SPFGRFK and KNLQDR, which are consistent with partial sequences of human NR1, and GVEDALVSLK, QPTVAGAPK, and NEVMSSK, which correspond to those of NR2A, NR2B and NR2C, respectively. Immunocytochemical analysis revealed that the bovine ovum was stained with the immunoglobulin G purified from the serum of a patient with anti-NMDAR encephalitis. Taken together, we propose that the normal ovum expresses NMDARs that have strong affinity for the disease-specific IgG. The presence of NMDARs in ova may help explain why young females without ovarian teratomas are also affected by anti-NMDAR encephalitis.


Staff workload and adverse events during mechanical ventilation in neonatal intensive care units.
To investigate a possible association between the intensity of staff workload and intermediate adverse events, such as accidental extubation, obstruction of the endotracheal tube, and accidental disconnection of the ventilator circuit, during neonatal mechanical ventilation in high-risk neonatal units. This prospective cohort study analyzed data of 543 newborns from public neonatal intensive care units (NICUs) in the city of São Luís, state of Maranhão, Northeastern Brazil, for 6 months, during which 136 newborns were submitted to mechanical ventilation in 1,108 shifts and were observed a total of 4,554 times. Adverse events occurred 117 times during this period. The associations between workload and adverse events were analyzed by means of generalized estimating equations. The adjustment variables were: birth weight, gender, maternity unit, Clinical Risk Index for Babies score, and care demand, the latter measured by the Northern Neonatal Network Scale. The larger the number of newborns classified by care demand (NCCD) per nurse and nursing technician, the more likely the occurrence of intermediate adverse events linked to mechanical ventilation. A number of NCCD > 22 per nurse (relative risk [RR] = 2.86) and > 4.8 per auxiliary nurse (RR = 3.41) was associated with a higher prevalence of intermediate adverse events. The workload of NICU professionals seems to interfere with the intermediate results of neonatal care and thus should be taken into consideration when evaluating NICU outcomes.


Allozyme analysis of two polymorphic enzymes in a natural population of Lecithochirium musculus.
Genetic variability among 112 individuals of Lecithochirium musculus from Anguilla anguilla from northwestern Spain was examined using allozyme analysis. Starch-gel electrophoresis was used to investigate the genetic variation in two polymorphic enzymes, glucose phosphate isomerase (GPI) and phosphoglucomutase (PGM). The banding patterns obtained for GPI were consistent with a dimeric structure for this enzyme and with single-locus control. The inferred genotype frequencies did not significantly differ as compared with those predicted by the Hardy-Weinberg equation. Two PGM loci were detected (Pgm-1 and Pgm-2). Pgm-1 was polymorphic and showed a striking departure from Hardy-Weinberg predictions (total absence of heterozygotes for alleles in high frequencies), raising the possibility of the existence of two reproductively isolated populations. However, other reasons for this observation are discussed.


A novel immunohistochemical scoring system reveals associations of C-terminal MET, ectodomain shedding, and loss of E-cadherin with poor prognosis in oral squamous cell carcinoma.
Using tissue microarrays, it was shown that membranous C-terminal MET immunoreactivity and ectodomain (ECD) shedding are associated with poor prognosis in oral cancer. Seen the potential diagnostic value, extrapolation of these results to whole-tissue sections was investigated. Because MET orchestrates epithelial-to-mesenchymal transition (EMT), the results were benchmarked to loss of E-cadherin, a readout for EMT known to be associated with poor prognosis. C-terminal MET, N-terminal MET, and E-cadherin immunoreactivities were examined on formalin-fixed paraffin-embedded parallel sections of 203 oral cancers using antibody clones D1C2, A2H2-3, and NCH-38. Interantibody and intra-antibody relations were examined using a novel scoring system, nonparametric distribution, and median tests. Survival analyses were used to examine the prognostic value of the observed immunoreactivities. Assessment of the three clones revealed MET protein status (no, decoy, transmembranous C-terminal positive), ECD shedding, and EMT. For C-terminal MET-positive cancers, D1C2 immunoreactivity is independently associated with poor overall survival (hazard ratio [HR] = 2.40; 95% confidence interval [CI] = 1.25 to 4.61; and P = 0.008) and disease-free survival (HR = 1.83; 95% CI = 1.07-3.14; P = 0.027). For both survival measures, this is also the case for ECD shedding (43.4%, with HR = 2.30; 95% CI = 1.38 to 3.83; and P = 0.001 versus HR = 1.87; 95% CI = 1.19-2.92; P = 0.006) and loss of E-cadherin (55.3%, with HR = 2.21; 95% CI = 1.30 to 3.77; and P = 0.004 versus HR = 1.90; 95% CI = 1.20-3.01; P = 0.007). The developed scoring system accounts for MET protein status, ECD shedding, and EMT and is prognostically informative. These findings may contribute to development of companion diagnostics for MET-based targeted therapy.


The registration of diagnostic versus planning fluorodeoxyglucose positron emission tomography/computed tomography in radiotherapy planning for non-small cell lung cancer.
Radiotherapy for non-small cell lung cancer (NSCLC) increasingly utilises fluorodeoxyglucose positron emission tomography/computed tomography (PET/CT) fusion. However, it is unknown whether a PET/CT scan conducted in the treatment position results in more accurate registration to the radiotherapy planning CT (rCT) than a diagnostic PET/CT scan. The aim of this study was to compare the accuracy of registration of the CT components of the planning PET/CT scan (pCT) and diagnostic PET/CT scan (dCT) scan with the rCT. Ten patients with stage I-III NSCLC underwent an rCT immediately followed by a planning PET/CT scan, both carried out with arms placed above the head and immobilisation in the treatment position. All previously underwent a diagnostic FDG PET/CT, which was carried out with the arms above the head, but without custom immobilisation. dCT and pCT were registered to the rCT using a rigid body mutual information algorithm. Four observers identified 12 anatomical points on each scan and differences in their absolute location were analysed. At the carina, the mean absolute error (MAE) for pCT-rCT compared with dCT-rCT was 4.37 versus 5.73 mm (P=0.028). However, there was no significant difference in the root mean squared error (RMSE) for that point. There were no significant differences in MAE or RMSE for all other anatomical points. The MAE for all points was 4.11 versus 4.15 mm (P=NS) and RMSE was 4.40 versus 4.48 mm for pCT-rCT compared with dCT-rCT (P=NS). There is an average of 4mm of misregistration when registering the CT components of PET/CT scans to the rCT for NSCLC. Using a rigid registration technique, the registration of a diagnostic PET/CT is as good as the registration of a planning PET/CT.


Transplantation for children with acute myeloid leukemia: a comparison of outcomes with reduced intensity and myeloablative regimens.
The safety and efficacy of reduced-intensity conditioning (RIC) regimens for the treatment of pediatric acute myeloid leukemia is unknown. We compared the outcome of allogeneic hematopoietic cell transplantation in children with acute myeloid leukemia using RIC regimens with those receiving myeloablative-conditioning (MAC) regimens. A total of 180 patients were evaluated (39 with RIC and 141 with MAC regimens). Results of univariate and multivariate analysis showed no significant differences in the rates of acute and chronic graft-versus-host disease, leukemia-free, and overall survival between treatment groups. The 5-year probabilities of overall survival with RIC and MAC regimens were 45% and 48%, respectively (P = .99). Moreover, relapse rates were not higher with RIC compared with MAC regimens (39% vs 39%; P = .95), and recipients of MAC regimens were not at higher risk for transplant-related mortality compared with recipients of RIC regimens (16% vs 16%; P = .73). After carefully controlled analyses, we found that in this relatively modest study population, the data supported a role for RIC regimens for acute myeloid leukemia in children undergoing allogeneic hematopoietic cell transplantation. The data also provided justification for designing a carefully controlled randomized clinical trial that examines the efficacy of regimen intensity in this population.


Single transmembrane domain insulin-like growth factor-II/mannose-6-phosphate receptor regulates central cholinergic function by activating a G-protein-sensitive, protein kinase C-dependent pathway.
The insulin-like growth factor-II/mannose-6-phosphate (IGF-II/M6P) receptor is a single-pass transmembrane glycoprotein that plays an important role in the intracellular trafficking of lysosomal enzymes and endocytosis-mediated degradation of IGF-II. However, its role in signal transduction after IGF-II binding remains unclear. In the present study, we report that IGF-II/M6P receptor in the rat brain is coupled to a G-protein and that its activation by Leu27IGF-II, an analog that binds rather selectively to the IGF-II/M6P receptor, potentiates endogenous acetylcholine release from the rat hippocampal formation. This effect is mediated by a pertussis toxin (PTX)-sensitive GTP-binding protein and is dependent on protein kinase Calpha (PKCalpha)-induced phosphorylation of downstream substrates, myristoylated alanine-rich C kinase substrate, and growth associated protein-43. Additionally, treatment with Leu27IGF-II causes a reduction in whole-cell currents and depolarization of cholinergic basal forebrain neurons. This effect, which is blocked by an antibody against the IGF-II/M6P receptor, is also sensitive to PTX and is mediated via activation of a PKC-dependent pathway. These results together revealed for the first time that the single transmembrane domain IGF-II/M6P receptor expressed in the brain is G-protein coupled and is involved in the regulation of central cholinergic function via the activation of specific intracellular signaling cascades.


Ex vivo activity of endoperoxide antimalarials, including artemisone and arterolane, against multidrug-resistant Plasmodium falciparum isolates from Cambodia.
Novel synthetic endoperoxides are being evaluated as new components of artemisinin combination therapies (ACTs) to treat artemisinin-resistant Plasmodium falciparum malaria. We conducted blinded ex vivo activity testing of fully synthetic (OZ78 and OZ277) and semisynthetic (artemisone, artemiside, artesunate, and dihydroartemisinin) endoperoxides in the histidine-rich protein 2 enzyme-linked immunosorbent assay against 200 P. falciparum isolates from areas of artemisinin-resistant malaria in western and northern Cambodia in 2009 and 2010. The order of potency and geometric mean (GM) 50% inhibitory concentrations (IC50s) were as follows: artemisone (2.40 nM) > artesunate (8.49 nM) > dihydroartemisinin (11.26 nM) > artemiside (15.28 nM) > OZ277 (31.25 nM) > OZ78 (755.27 nM). Ex vivo activities of test endoperoxides positively correlated with dihydroartemisinin and artesunate. The isolates were over 2-fold less susceptible to dihydroartemisinin than the artemisinin-sensitive P. falciparum W2 clone and showed sensitivity comparable to those with test endoperoxides and artesunate, with isolate/W2 IC50 susceptibility ratios of <2.0. All isolates had P. falciparum chloroquine resistance transporter mutations, with negative correlations in sensitivity to endoperoxides and chloroquine. The activities of endoperoxides (artesunate, dihydroartemisinin, OZ277, and artemisone) significantly correlated with that of the ACT partner drug, mefloquine. Isolates had mutations associated with clinical resistance to mefloquine, with 35% prevalence of P. falciparum multidrug resistance gene 1 (pfmdr1) amplification and 84.5% occurrence of the pfmdr1 Y184F mutation. GM IC50s for mefloquine, lumefantrine, and endoperoxides (artesunate, dihydroartemisinin, OZ277, OZ78, and artemisone) correlated with pfmdr1 copy number. Given that current ACTs are failing potentially from reduced sensitivity to artemisinins and partner drugs, newly identified mutations associated with artemisinin resistance reported in the literature and pfmdr1 mutations should be examined for their combined contributions to emerging ACT resistance.


Molecular Requirements of High-Fidelity Replication-Competent DNA Backbones for Orthogonal Chemical Ligation.
The molecular properties of the phosphodiester backbone that made it the evolutionary choice for the enzymatic replication of genetic information are not well understood. To address this, and to develop new chemical ligation strategies for assembly of biocompatible modified DNA, we have synthesized oligonucleotides containing several structurally and electronically varied artificial linkages. This has yielded a new highly promising ligation method based on amide backbone formation that is chemically orthogonal to CuAAC "click" ligation. A study of kinetics and fidelity of replication through these artificial linkages by primer extension, PCR, and deep sequencing reveals that a subtle interplay between backbone flexibility, steric factors, and ability to hydrogen bond to the polymerase modulates rapid and accurate information decoding. Even minor phosphorothioate modifications can impair the copying process, yet some radical triazole and amide DNA backbones perform surprisingly well, indicating that the phosphate group is not essential. These findings have implications in the field of synthetic biology.


Immunomodulatory effects of (n-3) fatty acids: putative link to inflammation and colon cancer.
Chronic inflammation and colorectal cancer are closely linked. Although the overall mechanisms of inflammation-associated gastrointestinal carcinogenesis are complex, it is clear that antiinflammatory therapy is efficacious against neoplastic progression and malignant conversion. From a dietary perspective, fish oil containing (n-3) polyunsaturated fatty acids (PUFAs) has antiinflammatory properties, but for years the mechanism has remained obscure. Of relevance to the immune system in the intestine, we showed that (n-3) PUFA feeding alters the balance between CD4+ T-helper (Th1 and Th2) subsets by directly suppressing Th1 cell development (i.e., clonal expansion). This is noteworthy because Th1 cells mediate inflammatory diseases and resistance to intracellular pathogens or allergic hypersensitivity, and Th2 cells mediate resistance to extracellular pathogens. Therefore, any changes induced by (n-3) PUFAs in T-cell subset balance and function are important because the outcome is expected to suppress the development of autoimmune diseases and possibly the occurrence of colon cancer. Precisely how the immunomodulatory effects of (n-3) PUFAs influence inflammation-associated colonic tumor development is the subject of an ongoing investigation.


Frequent mutations in the MITF pathway in melanoma.
Microphthalmia-associated transcription factor (MITF) is involved in melanocyte cell development, pigmentation and neoplasia. To determine whether MITF is somatically mutated in melanoma, we compared the sequence of MITF from primary and metastatic lesions to patient-matched normal DNA. In the 50 metastatic melanoma tumor lines analysed, we discovered four samples that had genomic amplifications of MITF and four that had MITF mutations in the regions encoding the transactivation, DNA binding or basic, helix-loop-helix domains. Sequence analysis for SOX10, a transcription factor, which both acts upstream of MITF and synergizes with MITF, identified an additional three samples with frameshift or nonsense mutations. Microphthalmia-associated transcription factor and SOX10 were found to be mutated in a mutually exclusive fashion, possibly suggesting disruption in a common genetic pathway. Taken together we found that over 20% of the metastatic melanoma cases had alterations in the MITF pathway. We show that the MITF pathway is also altered in primary melanomas: 2/26 demonstrated mutations in MITF and 6/55 demonstrated mutations in SOX10. Our findings suggest that altered MITF function during melanomagenesis can be achieved by MITF amplification, MITF single base substitutions or by mutation of its regulator SOX10.


Faith's wager: How religion deters gambling.
The relationship between religion and gambling has only rarely been investigated in sociology and related fields. Prior studies have found that religion, broadly defined, deters gambling, with different religious traditions exhibiting varying degrees of deterrence. Our study, a quantitative analysis of a recent representative sample of U.S. adults, theorizes and tests how three different dimensions of religion affect three distinct forms of gambling. Religious tradition and religious service attendance are found to reduce the likelihood of casino gambling and lottery play; while religious salience is the only dimension that constrains online gambling. We argue that these findings reflect variation in the social visibility, time intensity, and broader legitimacy associated with gambling forms, and that this variation is crucial for understanding the deterring effects of faith.


The 2002 Denali fault earthquake, Alaska: a large magnitude, slip-partitioned event.
The MW (moment magnitude) 7.9 Denali fault earthquake on 3 November 2002 was associated with 340 kilometers of surface rupture and was the largest strike-slip earthquake in North America in almost 150 years. It illuminates earthquake mechanics and hazards of large strike-slip faults. It began with thrusting on the previously unrecognized Susitna Glacier fault, continued with right-slip on the Denali fault, then took a right step and continued with right-slip on the Totschunda fault. There is good correlation between geologically observed and geophysically inferred moment release. The earthquake produced unusually strong distal effects in the rupture propagation direction, including triggered seismicity.


Reactive Oxygen Species Regulate Both Priming and Established Arthritis, but with Different Mechanisms.
Neutrophil cytosolic factor 1 (NCF1) is a key regulatory component of the phagocytic NOX2 complex, which produces reactive oxygen species (ROS). Polymorphism of the Ncf1 gene is associated with increased arthritis severity. In this study, we generated targeted Ncf1 knock-in mice with inducible Ncf1 expression and determined the critical time window during which the NOX2-derived ROS protect the mice from arthritis. Targeted Ncf1 knock-in mice lacked NOX2-derived ROS, and in vivo allelic conversion of Ncf1 by the CreERT2 recombinase led to full protein expression and ROS production within 10 days. Mice in which Ncf1 had been activated before immunization with type II collagen (CII) developed only mild clinical symptoms of collagen-induced arthritis (CIA), whereas the ROS-deficient littermates had severe arthritis. The functional Ncf1 restricted the expansion of IL-17A-producing T cells specific for the immunodominant CII peptide. When the Ncf1 gene was activated after the priming phase, Ncf1-dependent protection from autoimmune arthritis was still observed, together with a reduced number of splenic monocytes but it was not associated with alterations in peptide-specific T cell response. The Ncf1-deficient mice expressed pronounced interferon signature, which could be normalized by conditional expression of Ncf1 and was also present in the Ncf1-mutated mouse during arthritis. Innovation and Conclusion: Ncf1 deficiency has been known to predispose to autoimmunity in both humans and rodents. Our in vivo results point to a regulatory role of NOX2-derived ROS not only during priming but also during the effector phase of CIA, most likely via different mechanisms. Antioxid. Redox Signal. 27, 1473-1490.