A standardized relative resource cost model for medical care: application to cancer control programs.
Medicare data represent 75% of aged and permanently disabled Medicare beneficiaries enrolled in the fee-for-service (FFS) indemnity option, but the data omit 25% of beneficiaries enrolled in Medicare Advantage health maintenance organizations (HMOs). Little research has examined how longitudinal patterns of utilization differ between HMOs and FFS. The Burden of Cancer Study developed and implemented an algorithm to assign standardized relative costs to HMO and Medicare FFS data consistently across time and place. Medicare uses 15 payment systems to reimburse FFS providers for covered services. The standardized relative resource cost algorithm (SRRCA) adapts these various payment systems to utilization data. We describe the rationale for modifications to the Medicare payment systems and discuss the implications of these modifications. We applied the SRRCA to data from four HMO sites and the linked Surveillance, Epidemiology, and End Results-Medicare data. Some modifications to Medicare payment systems were required, because data elements needed to categorize utilization were missing from both data sources. For example, data were not available to create episodes for home health services received, so we assigned costs per visit based on visit type (nurse, therapist, and aide). For inpatient utilization, we modified Medicare's payment algorithm by changing it from a flat payment per diagnosis-related group to daily rates for diagnosis-related groups to differentiate shorter versus longer stays. The SRRCA can be used in multiple managed care plans and across multiple FFS delivery systems within the United States to create consistent relative cost data for economic analyses. Prior to international use of the SRRCA, data need to be standardized.


Preliminary Findings of a Dedicated Ocular Myasthenia Gravis Rating Scale: The OMGRate.
There is a pressing need for a robust rating scale for ocular myasthenia gravis (OMG). Rating scales for myasthenia gravis (MG) research have a predominant focus on generalised disease. We present results of the first dedicated rating scale for OMG: the ocular myasthenia gravis rating scale (OMGRate). The OMGRate was developed through an international collaboration between neuromuscular and neuro-ophthalmology experts in OMG. It comprises two components: a physician- examination (OMGRate-e) and a patient questionnaire (OMGRate-q).. The OMGRate was prospectively validated in patients attending a neuro-ophthalmology clinic from April 2017 to October 2018. External validity and reliability of OMGRate were evaluated using validated MG rating scales: the Myasthenia Gravis Composite (MGC), the Myasthenia Gravis Quality of Life (MG-QOL), and the ocular component from the Myasthenia Gravis Impairment Index questionnaire (MGII). Two hundred and eleven assessments were completed in 104 patients (67 males, mean age 55 y, range 18-86 y). There was very good external validity of the OMGRate: good correlation between OMGRate-e and MGC (r = 0.64, 95% confidence intervals [CI] 0.54-0.74, p < .0001); excellent correlation between OMGRate-q and MGII (r = 0.85, 95% CI 0.78-0.91, p < .0001) and good correlation between OMGRate and MG-QOL (r = 0.68, 95% CI 0.60-0.77, p < .0001). A higher correlation of OMGRate and MG-QOL compared with MGC and MG-QOL (r = 0.47, 95% CI 0.34-0.59, p < .0001) suggests that OMGRate is better able to capture significant QOL information in patients with OMG. It had excellent reliability with an intraclass correlation coefficient of 0.83 (95% CI 0.67-0.92). Feedback from examiners and patients indicated that the OMGRate was easy to use. In conclusion, OMGRate is an easy-to-use, valid and reliable rating scale for monitoring the severity of OMG.


Effect of Renal Function on Dosing of Non-Vitamin K Antagonist Direct Oral Anticoagulants Among Patients With Nonvalvular Atrial Fibrillation.
Non-vitamin K antagonist direct oral anticoagulants (DOACs) are fixed-dose regimens indicated for stroke prevention in nonvalvular atrial fibrillation (NVAF) patients. Dose adjustment is necessary among patients with renal insufficiency to optimize efficacy and safety. To assess DOAC dosing appropriateness and its effect on clinical outcomes in NVAF patients. Adult NVAF patients with ≥1 DOAC pharmacy claim (January 1, 2013, to December 31, 2014), continuous enrollment for ≥12 months post-index DOAC claim, and documented creatinine clearance within 3 months preindex date in the Optum/Humedica SmartFile database were eligible. DOAC dosage was classified as inappropriate or appropriate by level of renal function, age, and body weight per US prescription information. Cox proportional models were used to assess the risks of bleeding and stroke associated with inappropriate DOAC dosage. Of the 388 eligible patients, 69 (17.8%) were inappropriately dosed, and rivaroxaban had the highest inappropriate dosing rate. Most inappropriately dosed patients were underdosed. Inappropriately dosed patients were more likely to be older, female, and have a body weight of ≤60 kg; they also had higher mean CHA2DS2-VASc and Charlson comorbidity index scores (all P < 0.05). Overtreated patients had a higher risk of bleeding (hazard ratio [HR] = 5.4; P = 0.006) than undertreated patients (HR = 3.1; P = 0.025) relative to appropriately dosed patients. However, no significant difference in stroke risk was observed, most likely because very few stroke events were observed in the study. Inappropriate dosing occurred among patients with normal and insufficient renal function. The consideration of clinical factors beyond renal function is necessary to reduce bleeding risk associated with DOAC therapy.


Experiences from pregnancy and childbirth related to female genital mutilation among Eritrean immigrant women in Sweden.
to explore Eritrean immigrant women's experiences of female genital mutilation (FGM) during pregnancy, childbirth and the postpartum period. qualitative study using an ethnographic approach. Data were collected via tape-recorded interviews. interviews in the Eritrean women's homes located in and around Uppsala, Sweden. 15 voluntary Eritrean immigrant women. Semi-structured interview and open-ended questions were used. The interviews were tape-recorded, transcribed verbatim and then analysed. six themes of experiences of FGM among Eritrean women during pregnancy and childbirth were identified. They are (1) fear and anxiety; (2) extreme pain and long-term complications; (3) health-care professionals' knowledge of circumcision and health-care system; (4) support from family, relatives and friends; (5) de-infibulation; and (6) decision against female circumcision of daughters. the Eritrean women had experiences of FGM and had suffered from its complications during pregnancy, childbirth and the postpartum period. Midwives and obstetricians should have competence in managing women with FGM, and they need increased understanding of cultural epistemology in order to be able to provide quality care to these women. At antenatal centres, circumcised women should be advised to de-infibulate before pregnancy. Special courses about anatomical differences should be offered to these women and their husbands. It is also important to inform them about Swedish law, which prohibits all forms of FGM.


Microbial Electrolytic Carbon Capture for Carbon Negative and Energy Positive Wastewater Treatment.
Energy and carbon neutral wastewater management is a major goal for environmental sustainability, but current progress has only reduced emission rather than using wastewater for active CO2 capture and utilization. We present here a new microbial electrolytic carbon capture (MECC) approach to potentially transform wastewater treatment to a carbon negative and energy positive process. Wastewater was used as an electrolyte for microbially assisted electrolytic production of H2 and OH(-) at the cathode and protons at the anode. The acidity dissolved silicate and liberated metal ions that balanced OH(-), producing metal hydroxide, which transformed CO2 in situ into (bi)carbonate. Results using both artificial and industrial wastewater show 80-93% of the CO2 was recovered from both CO2 derived from organic oxidation and additional CO2 injected into the headspace, making the process carbon-negative. High rates and yields of H2 were produced with 91-95% recovery efficiency, resulting in a net energy gain of 57-62 kJ/mol-CO2 captured. The pH remained stable without buffer addition and no toxic chlorine-containing compounds were detected. The produced (bi)carbonate alkalinity is valuable for wastewater treatment and long-term carbon storage in the ocean. Preliminary evaluation shows promising economic and environmental benefits for different industries.


1H, 13C, and 15N backbone chemical shift assignments of the apo and the ADP-ribose bound forms of the macrodomain of SARS-CoV-2 non-structural protein 3b.
The SARS-CoV-2 genome encodes for approximately 30 proteins. Within the international project COVID19-NMR, we distribute the spectroscopic analysis of the viral proteins and RNA. Here, we report NMR chemical shift assignments for the protein Nsp3b, a domain of Nsp3. The 217-kDa large Nsp3 protein contains multiple structurally independent, yet functionally related domains including the viral papain-like protease and Nsp3b, a macrodomain (MD). In general, the MDs of SARS-CoV and MERS-CoV were suggested to play a key role in viral replication by modulating the immune response of the host. The MDs are structurally conserved. They most likely remove ADP-ribose, a common posttranslational modification, from protein side chains. This de-ADP ribosylating function has potentially evolved to protect the virus from the anti-viral ADP-ribosylation catalyzed by poly-ADP-ribose polymerases (PARPs), which in turn are triggered by pathogen-associated sensing of the host immune system. This renders the SARS-CoV-2 Nsp3b a highly relevant drug target in the viral replication process. We here report the near-complete NMR backbone resonance assignment (1H, 13C, 15N) of the putative Nsp3b MD in its apo form and in complex with ADP-ribose. Furthermore, we derive the secondary structure of Nsp3b in solution. In addition, 15N-relaxation data suggest an ordered, rigid core of the MD structure. These data will provide a basis for NMR investigations targeted at obtaining small-molecule inhibitors interfering with the catalytic activity of Nsp3b.


Current evaluation of cardiac stab wounds.
Patients with penetrating cardiac injuries may be stable or only mildly shocked, especially if the laceration has sealed off and the patient has been aggressively resuscitated. Clinical signs, chest roentgenograms, pericardiocentesis, and subxiphoid window are not always helpful in establishing the diagnosis. We reflect on the current evaluation based on 128 patients. There were four groups of patients, ranging from lifeless (group I) to stable (group IV). Patients in groups I and II were prepared immediately for operation. Those in groups III and IV were often investigated further (chest roentgenogram and cardiac ultrasound). Mortality was 8%. Significant findings were a precordial stab, central venous pressure of more than 15 cm of water, one or more clinical signs of tamponade, and initial shock. Cardiac ultrasound was performed in 5 patients in group II (15%), 14 patients in group III (48%), and 37 patients in group IV (86%). There were no false positives, and 6 false negatives (11%). Thirty-one patients (24%) had clotted lacerations. There were no negative sternotomies. Efficient fluid resuscitation and rapid confirmation of diagnosis with cardiac ultrasound should decrease mortality. Stable patients with a precordial wound should undergo cardiac ultrasound or echocardiogram. Diagnosis may be reliably confirmed in these patients whose clinical signs often fluctuate (or rapidly deteriorate).


Cloning and characterization of guinea pig CXCR1.
IL-8/CXCL8 plays a critical role in the trafficking and activation of neutrophils via its receptors, CXCR1 and CXCR2, in humans. CXCR1 is highly selective for IL-8, whereas CXCR2 is activated by all CXC chemokines with an ELR motif. In mice and rats, neither IL-8 nor CXCR1 is present, making it difficult to evaluate the in vivo roles of the IL-8/CXCR1 interactions. We previously demonstrated the presence of IL-8 in the guinea pig (gp), suggesting that its specific receptor CXCR1 is also present in this species. Here, we obtained two gp genomic DNA clones, clones 8 and 10, coding for the potential orthologues of CXCR1 and CXCR2, respectively. Transcripts for these genes were expressed in neutrophils, but not in macrophages. Functionally, both gp and human (h) IL-8 induced cell migration and ERK phosphorylation in HEK 293 cells expressing either receptor, whereas hGRO activated only cells expressing the clone 10 protein, confirming that clone 8 indeed coded for gpCXCR1. 125I-labeled hIL-8 bound to gpCXCR1 and addition of unlabeled hIL-8 completely abolished the binding; however, unlabeled gpIL-8 failed to compete against 125I-labeled hIL-8, strongly suggesting that the avidity of hIL-8 to gpCXCR1 is higher than that of gpIL-8. Identification and characterization of CXCR1 in the guinea pig will allow us to use this small animal model to evaluate the role of the IL-8/CXCR1 interactions and to examine the efficacy of CXCR1 antagonists in vivo.


Stage-dependent distribution of calreticulin in oocytes of the frog, Rana rugosa.
We studied immunohistologically the distribution of a Ca2(+)-binding protein, calreticulin (CLT), at different stages of growing oocytes in the frog, Rana rugosa. Northern blot analysis showed that the CLT gene expression in gonads of metamorphosing tadpoles was low, but was extremely strong in the ovary, but not in the testis, of 2-month post-metamorphosis frogs, followed by decline to a lower level in adult frogs. On the contrary, the beta-actin gene expression did not increase in the same ovary. As for the distribution of CLT protein, a weak immunostaining was observed in indifferent gonads of tadpoles at stage I. The CLT distribution in oocytes from stage A to F was stage-dependent. In addition, Western blot analysis revealed that the CLT level was low in gonads of tadpoles at stage I, but increased at stage XVI. It still increased in the ovary of frogs 2 months after metamorphosis, and then dropped to a lower level in adult frogs. The results indicate that CLT gene expression occurs in the early stage of growing oocytes, and that CLT is synthesized actively in oocytes in the ovary of frogs after metamorphosis. Based on these findings, the role of CLT is discussed.


The CENP-T C-terminus is exclusively proximal to H3.1 and not to H3.2 or H3.3.
The kinetochore proteins assemble onto centromeric chromatin and regulate DNA segregation during cell division. The inner kinetochore proteins bind centromeres while most outer kinetochore proteins assemble at centromeres during mitosis, connecting the complex to microtubules. The centromere-kinetochore complex contains specific nucleosomes and nucleosomal particles. CENP-A replaces canonical H3 in centromeric nucleosomes, defining centromeric chromatin. Next to CENP-A, the CCAN multi-protein complex settles which contains CENP-T/W/S/X. These four proteins are described to form a nucleosomal particle at centromeres. We had found the CENP-T C-terminus and the CENP-S termini next to histone H3.1 but not to CENP-A, suggesting that the Constitutive Centromere-Associated Network (CCAN) bridges a CENP-A- and a H3-containing nucleosome. Here, we show by in vivo FRET that this proximity between CENP-T and H3 is specific for H3.1 but neither for the H3.1 mutants H3.1(C96A) and H3.1(C110A) nor for H3.2 or H3.3. We also found CENP-M next to H3.1 but not to these H3.1 mutants. Consistently, we detected CENP-M next to CENP-S. These data elucidate the local molecular neighborhood of CCAN proteins next to a H3.1-containing centromeric nucleosome. They also indicate an exclusive position of H3.1 clearly distinct from H3.2, thus documenting a local, and potentially also functional, difference between H3.1 and H3.2.


Maxillary involvement in central craniofacial fractures with associated head injuries.
From January 1983 through December 1990, we treated 703 patients with central craniofacial fractures. Among them 177 had maxillary fractures, with or without frontonasoethmoidal involvement. Of the 177, 151 patients were diagnosed in the acute stage. Using the Glasgow Coma Scale, these 151 patients were divided into three groups: group I (GCS score 13-15), 100 patients; group II (GCS score 9-12), 33 patients; group III (GCS score 3-8), 18 patients. The patterns of facial fractures were analyzed and surgically corrected after the following average postadmission lags: group I, 7.1 days; group II, 11.5 days; group III, 13.3 days. The average hospital stay for each group was: group I, 15.6 days; group II, 19.5 days; and group III, 27.2 days. The mean follow-up period was 12 months. The physiologic, functional and esthetic outcomes were assessed. Regardless of initial head injury severity, the outcomes in each group showed almost no statistically significant differences, and unnecessary extended hospital stays may be avoided by early surgical management.


Causality in scale space as an approach to change detection.
Kernel density estimation and kernel regression are useful ways to visualize and assess the structure of data. Using these techniques we define a temporal scale space as the vector space spanned by bandwidth and a temporal variable. In this space significance regions that reflect a significant derivative in the kernel smooth similar to those of SiZer (Significant Zero-crossings of derivatives) are indicated. Significance regions are established by hypothesis tests for significant gradient at every point in scale space. Causality is imposed onto the space by restricting to kernels with left-bounded or finite support and shifting kernels forward. We show that these adjustments to the methodology enable early detection of changes in time series constituting live surveillance systems of either count data or unevenly sampled measurements. Warning delays are comparable to standard techniques though comparison shows that other techniques may be better suited for single-scale problems. Our method reliably detects change points even with little to no knowledge about the relevant scale of the problem. Hence the technique will be applicable for a large variety of sources without tailoring. Furthermore this technique enables us to obtain a retrospective reliable interval estimate of the time of a change point rather than a point estimate. We apply the technique to disease outbreak detection based on laboratory confirmed cases for pertussis and influenza as well as blood glucose concentration obtained from patients with diabetes type 1.


The Antioxidant Potential of Azadirachta indica Ameliorates Cardioprotection Following Diabetic Mellitus-Induced Microangiopathy.
Cardiac complications associated with diabetes mellitus have become major cause of concern. Antidiabetic drugs, with varied mode of action, are although available, apprehensions exist for their limited action or side effects upon prolonged use. Efforts are therefore inclined toward finding other alternatives. The present study was, thus, undertaken to evaluate the cardioprotective effect of Azadirachta indica (AI) on microangiopathic changes in rat model of diabetes. Diabetes was induced in male rats by single intraperitoneal injection of streptozotocin (60 mg/kg body weight). Seven days after glucose levels are stabilized, aqueous leaf extract of AI (ALE) (600 mg/kg(1) body weight) was administered orally to diabetic animals every day for 7 days. High blood glucose characterizing diabetes in these animals was found to show increased lipid peroxidation (LPO), altered antioxidant biomarkers together with microangiopathic alterations. The treatment of diabetic rats with ALE reduced the levels of blood glucose, LPO, and restored the activities of antioxidant enzyme. Light and transmission electron microscopic analysis revealed reduced necrotic areas and inflammation in tissue architecture of ALE treated heart in comparison to untreated diabetic group. AI provides cardioprotection by ameliorating oxidative stress in rat model of diabetic mellitus. The streptozotocin (STZ) treatment (60 mg/kg body weight) to animals induced diabetic changes such as elevated blood glucose levels, decreased body weight, altered lipid profiles together with development of proxidant state evidenced by elevated levels of lipid peroxidation (LPO), depletion in reduced glutathione (GSH) levels and altered antioxidant enzymes with consequent microangiopathic alterations in heart tissue evinced by localization of necrotic and inflamed areas in heart tissueThe treatment of animals with Azadirachta indica leaf extract (ALE) (600 mg/kg body weight) post-STZ treatment significantly reversed the adverse effects witnessed by normalized blood glucose levels, improvement in reduced body weight and stabilized lipid profilesFurther, ALE treatment also significantly reduced the LPO indices, improvement in GSH content and restoration of antioxidant enzyme activities, suggesting antioxidatant potential of ALEThe microangiopathic changes in the heart tissue consequent to induction of diabetes and oxidative stress by STZ as reiterated through light microscopy and transmission electron microscopy were found to be reversed by ALE treatment. These observations pointed toward cardiopreventive effects of ALE following microangiopathic changes as seen following induction of diabetes mellitus. Abbreviations used: AI: Azadirachta indica, ALE: Azadirachta indica Leaves Extract. STZ: Streptozotocin, LPO Lipid per oxidation, GSH: Glutathione, GSSG: Glutathione disulphide, SOD: Superoxide dismutase, GP: Glutathione peroxidase, GR: Glutathione reductase.


Ingenol mebutate: A novel topical drug for actinic keratosis.
The global incidence of non-melanoma skin cancer is rising. Significant morbidity leading to unacceptable cosmetic outcomes and/or functional impairment is a major concern. Search for non-surgical, non-invasive and tissue-sparing treatment modalities has led to development of new therapeutic agents. Actinic keratoses (AK) are one part of a continuous spectrum of benign sun damage to squamous cell carcinoma (SCC). Although it is not possible to predict which AK might progress to SCC, the presence of AK is a biomarker of risk for patients and must be treated to avoid possible morbidity and mortality. Ingenol mebutate is a novel topical drug from the latex sap of a plant-Euphorbia peplus that acts by chemoablative and immunostimulatory properties. Clinical studies have proven it to be safe and efficacious, leading to FDA approval of this chemotherapeutic agent for field therapy of AK in 2012. Current topical agents for field therapy of AK must be applied for weeks, whereas ingenol needs to be applied for three days. Ingenol offers a new therapeutic option that is convenient, safe, effective, acceptable and well-tolerated.


Antimutagenic activity of green tea and black tea extracts studied in a dynamic in vitro gastrointestinal model.
An in vitro gastrointestinal model, which simulates the conditions in the human digestive tract, was used to determine potential antimutagenic activity of extracts of black tea and green tea. In this paper, results are presented on the availability for absorption of potential antimutagenic compounds present in tea and on the influence of the food matrix on this activity. Between 60 and 180min after the tea was introduced into the model, antimutagenic activity was recovered from the jejunal compartment by means of dialysis: the dialysate appeared to inhibit the mutagenicity of the food mutagen MeIQx in the direct plate assay with Salmonella typhimurium (Ames test). The maximum inhibition was measured at 2h after the start of the experiment and was comparable for black tea and green tea extract. To determine the influence of food matrices on the antimutagenic activity of tea, the model was loaded with black tea together with milk or a homogenized standard breakfast. The maximum inhibition observed with black tea was reduced by 22, 42 and 78% in the presence of whole milk, semi-skimmed milk, and skimmed milk, respectively. Whole milk and skimmed milk abolished the antimutagenic activity of green tea by more than 90%; for semi-skimmed milk the inhibition was more than 60%. When a homogenized breakfast was added into the model together with the black tea extract, the antimutagenic activity was completely eliminated. When tea and MeIQx were added together into the digestion model, MeIQx mutagenicity was efficiently inhibited, with green tea showing a slightly stronger antimutagenic activity than black tea. In this case, the addition of milk had only a small inhibiting effect on the antimutagenicity. Antioxidant capacity and the concentration of catechins were also measured in the jejunal dialysates. The reduction in antimutagenic activity corresponded with reduction in antioxidant capacity and with a decrease of concentration of three catechins, viz. catechin, epigallocatechin gallate and epigallocatechin. The in vitro gastrointestinal model appears to be a useful tool to study the antimutagenicity of food components.


The Effect of Hydrogen on Martensite Transformations and the State of Hydrogen Atoms in Binary TiNi-Based Alloy with Different Grain Sizes.
The analysis presented here shows that in B2-phase of Ti49.1Ni50.9 (at%) alloy, hydrogenation with further aging at room temperature decreases the temperatures of martensite transformations and then causes their suppression, due to hydrogen diffusion from the surface layer of specimens deep into its bulk. When hydrogen is charged, it first suppresses the transformations B2↔B19' and R↔B19' in the surface layer, and when its distribution over the volume becomes uniform, such transformations are suppressed throughout the material. The kinetics of hydrogen redistribution is determined by the hydrogen diffusion coefficient DH, which depends on the grain size. In nanocrystalline Ti49.1Ni50.9 (at%) specimens, DH is three times greater than its value in coarse-grained ones, which is likely due to the larger free volume and larger contribution of hydrogen diffusion along grain boundaries in the nanocrystalline material. According to thermal desorption spectroscopy, two states of hydrogen atoms with low and high activation energies of desorption exist in freshly hydrogenated Ti49.1Ni50.9 (at%) alloy irrespective of the grain size. On aging at room temperature, the low-energy states disappear entirely. Estimates by the Kissinger method are presented for the binding energy of hydrogen in the two states, and the nature of these states in binary hydrogenated TiNi-based alloys is discussed.


Towards High-Quality Visualization of Superfluid Vortices.
Superfluidity is a special state of matter exhibiting macroscopic quantum phenomena and acting like a fluid with zero viscosity. In such a state, superfluid vortices exist as phase singularities of the model equation with unique distributions. This paper presents novel techniques to aid the visual understanding of superfluid vortices based on the state-of-the-art non-linear Klein-Gordon equation, which evolves a complex scalar field, giving rise to special vortex lattice/ring structures with dynamic vortex formation, reconnection, and Kelvin waves, etc. By formulating a numerical model with theoretical physicists in superfluid research, we obtain high-quality superfluid flow data sets without noise-like waves, suitable for vortex visualization. By further exploring superfluid vortex properties, we develop a new vortex identification and visualization method: a novel mechanism with velocity circulation to overcome phase singularity and an orthogonal-plane strategy to avoid ambiguity. Hence, our visualizations can help reveal various superfluid vortex structures and enable domain experts for related visual analysis, such as the steady vortex lattice/ring structures, dynamic vortex string interactions with reconnections and energy radiations, where the famous Kelvin waves and decaying vortex tangle were clearly observed. These visualizations have assisted physicists to verify the superfluid model, and further explore its dynamic behavior more intuitively.


Mortality in early inflammatory polyarthritis: cardiovascular mortality is increased in seropositive patients.
To determine the degree and causes of any excess mortality observed during the early years of inflammatory polyarthritis (IP). Between 1990 and 1994, a total of 1,236 patients were registered with the Norfolk Arthritis Register, a primary care-based inception cohort. All patients were tracked on the National Health Service Central Register for notification of death. The vital status of each patient was determined as of December 31, 1999. Causes of death were coded according to the International Classification of Diseases, Ninth Revision. Expected death rates were calculated using annual death rates for the Norfolk population. Standardized mortality ratios (SMRs) were calculated for all IP patients and for the subgroups of patients who did and did not satisfy the American College of Rheumatology (ACR) 1987 criteria for rheumatoid arthritis (RA) at baseline, as well as for the subgroups who were and were not rheumatoid factor (RF) positive at baseline. By December 31, 1999, 160 patients (13%; 79 women and 81 men) had died. The median duration of followup in the entire cohort was 6.9 years. Mortality rates were not significantly increased in the entire group of patients with IP or in the subgroup who met the ACR 1987 criteria for RA at baseline. In contrast, RF-positive patients had an increased rate of death from all causes (SMR in men 1.51, in women 1.41). Cardiovascular disease was the most common cause of death. The majority of the excess mortality in the RF-positive patients could be attributed to cardiovascular causes (SMR in men 1.34, in women 2.02). Excess mortality in the early years of IP is confined to patients who are seropositive for RF. While excess cardiovascular mortality has been described in patients with established RA, this is the first report of premature death from heart disease in the early years of IP.


Lubrication of circumcision site for prevention of meatal stenosis in children younger than 2 years old.
Circumcision is one of the most common surgical operations throughout the world, and meatal stenosis is one its late complications. We evaluated the topical use of a lubricant jelly after circumcision in boys in order to reduce the risk of meatal stenosis. A randomized control trial was performed, in which 2 groups of boys younger the 2 years old underwent circumcision according to the sleeve method. The parents in the study group were instructed to use petroleum jelly on the circumcision site after each diaper change for 6 months. In the control group, no topical medication was used. The children were followed up regularly and evaluated for meatal stenosis, bleeding, infection, and recovery time. A total of 197 boys in each group completed the study. None of the children in the study group but 13 (6.6%) in the control group developed meatal stenosis (P < .001). Infection of the circumcision site was seen in 3 (1.5%) and 23 (11.7%) children of the lubricant and control groups, respectively (P < .001), and bleeding was seen in 6 (3.0%) and 37 (18.8%), respectively (P < .001). The mean time of recovery in the lubricant group was 3.8 +/- 1.2 days, while it was 6.9 +/- 4.2 days in the control group (P = .03). Based on the findings of this study, it seems logical to use a lubricant jelly for reducing postcircumcision meatal stenosis and other complications.


Endothelin-receptor antagonists in arterial hypertension: further indications?
Endothelin-1 exerts vasoactive, pro-inflammatory, hypertrophic, and profibrotic properties on the heart, kidney, and blood vessels. Hence, endothelin-receptor antagonists hold the potential to reduce blood pressure and to prevent complications of hypertension, atherosclerosis, and diabetes through blood pressure-independent effects on cardiovascular growth, inflammation, and fibrosis. These potentially important effects of endothelin antagonism may contribute to its therapeutic potential in hypertension and other cardiovascular disorders, including chronic renal failure and diabetes. First clinical trial evidence demonstrates a moderate reduction in blood pressure in studies of patients with mild-to-moderate essential hypertension and patients with resistant hypertension. Future large-scale randomized clinical trials will provide more insight into whether the blood-pressure reduction and promising pleiotropic effects observed with several members of this novel class of drugs, which are already established therapy in pulmonary hypertension, will translate into clinical benefit in patients with arterial hypertension.


Patients with FLT3-mutant AML needed to enroll on FLT3-targeted therapeutic clinical trials.
We sought to identify the total number of therapeutic trials targeting FLT3-mutant acute myeloid leukemia (AML) to estimate the number of patients needed to satisfy recruitment when compared with the incidence of this mutation in the US AML population. A systematic review of all therapeutic clinical trials focusing on adult FLT3-mutated AML was conducted from 2000 to 2017. An updated search was performed using ClinicalTrials.gov for trials added between October 2017 and December 2018. Analysis was performed for ClinicalTrials.gov search results from 2000 to 2017 to provide descriptive estimates of discrepancies between anticipated clinical trial enrollment using consistently cited rates of adult participation of 1%, 3%, and 5%, as well as 10% participation identified by the American Society of Clinical Oncology in 2008. Twenty-five pharmaceutical or biological agents aimed at treating FLT3-mutant AML were identified. Pharmaceutical vs cooperative group/nonprofit support was 2.3:1, with 30 different pharmaceutical collaborators and 13 cooperative group/nonprofit collaborators. The number of patients needed to satisfy study enrollment begins to surpass the upper bound of estimated participation in 2010, noticeably surpassing projected participation rates between 2015 and 2016. The number of patients needed to satisfy study enrollment surpasses 3% and 5% rates of historical participation for US-only trials in 2017. We estimate that 15% of all US patients with FLT3-mutant AML would have to enroll in US and internationally accruing trials to satisfy requirements in 2017, or approximately 3 times the upper level of historical participation rates in the United States. The current clinical trial agenda in this space requires high percentage enrollment for sustainability.


The effect of suprapubic catheterization versus transurethral catheterization after abdominal surgery on urinary tract infection: a randomized controlled trial.
Transurethral catheterization is generally associated with a higher incidence of urinary tract infections than suprapubic catheterization; however, suprapubic catheterization is associated with other disadvantages such as higher costs and a more difficult technique, and at the moment there is no consensus about the use of both catheter systems. Therefore, a prospective randomized study was performed to investigate the effects of suprapubic catheterization and transurethral catheterization in patients undergoing surgery on the incidence of urinary tract infections and patient satisfaction. Patients who underwent an elective laparotomy were randomized and received a suprapubic or transurethral catheter. The primary end point was urinary tract infection. Other parameters of urinary tract infection, as well as duration of catheterization, hospital stay, and number of recatheterizations and of relaparotomies were monitored. Treatment 'per protocol' was also analyzed after exclusion of patients receiving another catheter than randomized for. Patients were asked for their satisfaction with the catheters and complaints during and after catheterization. 165 patients were eligible, of whom 19 patients had to be excluded. 75 patients were allocated to receive the suprapubic catheter and 71 the transurethral catheter. There was no difference in the incidence of a urinary tract infection between the suprapubic group (n = 9/75; 12%) and the transurethral group (n = 8/71; 11%). Most patients (6/9) who developed a urinary tract infection in the suprapubic group, however, underwent recatheterization because of postoperative complications/sepsis and relaparotomy. The incidence of urinary tract infections in patients who received a suprapubic catheter and not a transurethral catheter was 3/59 (5%). The patients did not differ with respect to satisfaction and complaints. Being a men, recatheterization and duration of catheterization are risk factors. The incidence of a urinary tract infection between a suprapubic catheter and a transurethral catheter in patients undergoing major surgery was not different. A potential advantage of the suprapubic catheter (reduction of urinary tract infections) is probably partly negated, because transurethral catheters were used if recatheterization was indicated during the postoperative stay or due to complications.


An in situ method for estimating cell survival in a solid tumor.
The response of the murine Meth-A fibrosarcoma (a methylcholanthrene-induced tumor) to single and fractionated doses of X-irradiation, actinomycin D chemotherapy, and/or concomitant local tumor hyperthermia was assayed with the use of an in situ method for estimating cell kill within a solid tumor. The cell survival assay was based on a standard curve plotting number of inoculated viable cells (10(1) to 10(7)) with and without radiation (10 kilorads)-inactivated homologous tumor cells (heavily irradiated) versus the time required for i.m. tumors to grow to 1.0 cu cm. The time for post-treatment tumors to grow to 1.0 cu cm was cross-referenced to the standard curve, and the number of surviving cells contributing to tumor regrowth was estimated. The resulting surviving fraction curves closely resemble those obtained with in vitro systems. The advantages and limitations of this technique are discussed as a method for evaluating treatment effectiveness.


Use of posttraumatic stress disorder to support an insanity defense.
The authors examine the allegation that the diagnosis of posttraumatic stress disorder (PTSD) is frequently abused in the legal system as the basis for a defense of not guilty by reason of insanity. Data for the investigation were drawn from a study of insanity pleas gathered from court records in 49 counties in eight states. Data on the 28 insanity plea defendants for whom PTSD was diagnosed before or immediately after trial were compared with data on 8,135 defendants whose insanity pleas were based on other diagnoses. Insanity pleas by defendants with diagnoses of PTSD constituted only 0.3% of the cases. There were few significant differences between the two groups on demographic variables, psychiatric histories, previous involvement in crime, or current charges. The defendants with PTSD were more likely to have been married, less likely to have been arrested as juveniles, and less likely to have been detained after trial. Contrary to previously expressed concerns, PTSD was infrequently associated with an insanity defense in the cases in this study. In the cases in which pleas based on PTSD were used, they were no more likely to succeed than pleas based on any other diagnosis. Defendants with PTSD-related insanity defenses differed little from other insanity defendants, contradicting the stereotype of the person who is driven by PTSD to commit crimes. The data do not support fears of widespread misuse of the diagnosis of PTSD in connection with the insanity defense.


Footwear and Elevated Heel Influence on Barbell Back Squat: A Review.
The back squat is one of the most effective exercises in strengthening the muscles of the lower extremity. Understanding the impact of footwear has on the biomechanics is imperative for maximizing the exercise training potential, preventing injury, and rehabilitating from injury. This review focuses on how different types of footwear affect the full-body kinematics, joint loads, muscle activity, and ground reaction forces in athletes of varying experience performing the weighted back squat. The literature search was conducted using three databases, and fourteen full-text articles were ultimately included in the review. The majority of these studies demonstrated that the choice of footwear directly impacts kinematics and kinetics. Weightlifting shoes were shown to decrease trunk lean and generate more plantarflexion relative to running shoes and barefoot lifting. Elevating the heel through the use of external squat wedges is popular method during rehabilitation and was shown to provide similar effects to weightlifting shoes. Additional research with a broader array of populations, particularly novice and female weightlifters, should be conducted to generalize the research results to nonathlete populations. Further work is also needed to characterize the specific effects of sole stiffness and heel elevation height on squatting mechanics.


Refining the management of relapsed or refractory follicular lymphoma.
In patients with follicular lymphoma, the prolonged clinical course consisting of multiple relapses is a fundamental challenge that requires clinicians to consider how to best balance treatment efficacy while minimizing toxicity and preserving quality of life. The treatment approaches and decisions regarding therapy are largely driven by the unique clinical features evident in each patient. The traditional treatment approaches for relapsed follicular lymphoma include chemoimmunotherapy regimens, targeted agents, radioimmunotherapy, and, occasionally, immunotherapy alone. The primary targeted agents used in the relapsed or refractory follicular lymphoma setting are the phosphatidylinositol 3-kinase (PI3K) inhibitors idelalisib, copanlisib, and duvelisib. PI3K inhibitors can have a significant toxicity profile. Radioimmunotherapy remains an underutilized option. The newest agent that has gained regulatory approval in the treatment of follicular lymphoma is tazemetostat, a methyltransferase inhibitor that inhibits and reduces the activity of EZH2. In June 2020, tazemetostat was approved by the US Food and Drug Administration (FDA) for the treatment of adult patients with relapsed or refractory follicular lymphoma who have tumors that are positive for an EZH2 mutation (as detected by an FDA-approved test) and who have received at least 2 prior systemic therapies, or patients who have no satisfactory alternative treatment options. Data from a phase 2 study demonstrated that tazemetostat can produce clinically meaningful and durable responses, with a favorable safety profile, in heavily pretreated patients with or without an EZH2 mutation.


The Collaborative Chronic Care Model for Mental Health Conditions: From Evidence Synthesis to Policy Impact to Scale-up and Spread.
Extensive evidence indicates that Collaborative Chronic Care Models (CCMs) improve outcome in chronic medical conditions and depression treated in primary care. Beginning with an evidence synthesis which indicated that CCMs are also effective for multiple mental health conditions, we describe a multistage process that translated this knowledge into evidence-based health system change in the US Department of Veterans Affairs (VA). In 2010, recognizing that there had been numerous CCM trials for a wide variety of mental health conditions, we conducted an evidence synthesis compiling randomized controlled trials of CCMs for any mental health condition. The systematic review demonstrated CCM effectiveness across mental health conditions and treatment venues. Cumulative meta-analysis and meta-regression further informed our approach to subsequent CCM implementation. In 2015, based on the evidence synthesis, VA Office of Mental Health and Suicide Prevention (OMHSP) adopted the CCM as the model for their outpatient mental health teams. In 2015-2018 we partnered with OMHSP to conduct a 9-site stepped wedge implementation trial, guided by insights from the evidence synthesis. In 2017 OMHSP launched an effort to scale-up and spread the CCM to additional VA medical centers. Seventeen facilitators were trained and 28 facilities engaged in facilitation. Evidence synthesis provided leverage for evidence-based policy change. This formed the foundation for a health care leadership/researcher partnership, which conducted an implementation trial and subsequent scale-up and spread effort to enhance adoption of the CCM, as informed by the evidence synthesis.


Targeted Temperature Management for 48 vs 24 Hours and Neurologic Outcome After Out-of-Hospital Cardiac Arrest: A Randomized Clinical Trial.
International resuscitation guidelines recommend targeted temperature management (TTM) at 33°C to 36°C in unconscious patients with out-of-hospital cardiac arrest for at least 24 hours, but the optimal duration of TTM is uncertain. To determine whether TTM at 33°C for 48 hours results in better neurologic outcomes compared with currently recommended, standard, 24-hour TTM. This was an international, investigator-initiated, blinded-outcome-assessor, parallel, pragmatic, multicenter, randomized clinical superiority trial in 10 intensive care units (ICUs) at 10 university hospitals in 6 European countries. Three hundred fifty-five adult, unconscious patients with out-of-hospital cardiac arrest were enrolled from February 16, 2013, to June 1, 2016, with final follow-up on December 27, 2016. Patients were randomized to TTM (33 ± 1°C) for 48 hours (n = 176) or 24 hours (n = 179), followed by gradual rewarming of 0.5°C per hour until reaching 37°C. The primary outcome was 6-month neurologic outcome, with a Cerebral Performance Categories (CPC) score of 1 or 2 used to define favorable outcome. Secondary outcomes included 6-month mortality, including time to death, the occurrence of adverse events, and intensive care unit resource use. In 355 patients who were randomized (mean age, 60 years; 295 [83%] men), 351 (99%) completed the trial. Of these patients, 69% (120/175) in the 48-hour group had a favorable outcome at 6 months compared with 64% (112/176) in the 24-hour group (difference, 4.9%; 95% CI, -5% to 14.8%; relative risk [RR], 1.08; 95% CI, 0.93-1.25; P = .33). Six-month mortality was 27% (48/175) in the 48-hour group and 34% (60/177) in the 24-hour group (difference, -6.5%; 95% CI, -16.1% to 3.1%; RR, 0.81; 95% CI, 0.59-1.11; P = .19). There was no significant difference in the time to mortality between the 48-hour group and the 24-hour group (hazard ratio, 0.79; 95% CI, 0.54-1.15; P = .22). Adverse events were more common in the 48-hour group (97%) than in the 24-hour group (91%) (difference, 5.6%; 95% CI, 0.6%-10.6%; RR, 1.06; 95% CI, 1.01-1.12; P = .04). The median length of intensive care unit stay (151 vs 117 hours; P < .001), but not hospital stay (11 vs 12 days; P = .50), was longer in the 48-hour group than in the 24-hour group. In unconscious survivors from out-of-hospital cardiac arrest admitted to the ICU, targeted temperature management at 33°C for 48 hours did not significantly improve 6-month neurologic outcome compared with targeted temperature management at 33°C for 24 hours. However, the study may have had limited power to detect clinically important differences, and further research may be warranted. clinicaltrials.gov Identifier: NCT01689077.


Rethinking benefit sharing in collaborative human genetic research from an Afrocommunitarian perspective.
This article draws on reflections about humanness, friendliness and partiality, in the writings of Afro-communitarians to develop principles for thinking critically about why benefit sharing, what may count as benefits within the context of human research in Africa and the limits of the obligation of benefit sharing. Suppose the thinking about humanness, friendliness, and partiality in Afro-communitarianism were the foundation of human genetic research in Africa, then, individuals who have contributed to research or borne its burden would benefit from its rewards. This is even more important if participants have pressing needs that researchers and/or research institutions can help ease. A failure to aid sample contributors and data providers in need when researchers and research institutions can-as well as an indifference to the serious needs of contributors-are failures to exhibit friendliness in the relevant ways. Finally, though providing benefits to contributors can be an important way of showing humanity to them, nonetheless, this obligation is not absolute and may be limited by the stronger obligation of shared experience-to advance science. Studies are still required to inquire how well these norms will work in practice and inform regulatory and legal frameworks.


Pain in experimental autoimmune encephalitis: a comparative study between different mouse models.
Pain can be one of the most severe symptoms associated with multiple sclerosis (MS) and develops with varying levels and time courses. MS-related pain is difficult to treat, since very little is known about the mechanisms underlying its development. Animal models of experimental autoimmune encephalomyelitis (EAE) mimic many aspects of MS and are well-suited to study underlying pathophysiological mechanisms. Yet, to date very little is known about the sensory abnormalities in different EAE models. We therefore aimed to thoroughly characterize pain behavior of the hindpaw in SJL and C57BL/6 mice immunized with PLP139-151 peptide or MOG35-55 peptide respectively. Moreover, we studied the activity of pain-related molecules and plasticity-related genes in the spinal cord and investigated functional changes in the peripheral nerves using electrophysiology. We analyzed thermal and mechanical sensitivity of the hindpaw in both EAE models during the whole disease course. Qualitative and quantitative immunohistochemical analysis of pain-related molecules and plasticity-related genes was performed on spinal cord sections at different timepoints during the disease course. Moreover, we investigated functional changes in the peripheral nerves using electrophysiology. Mice in both EAE models developed thermal hyperalgesia during the chronic phase of the disease. However, whereas SJL mice developed marked mechanical allodynia over the chronic phase of the disease, C57BL/6 mice developed only minor mechanical allodynia over the onset and peak phase of the disease. Interestingly, the magnitude of glial changes in the spinal cord was stronger in SJL mice than in C57BL/6 mice and their time course matched the temporal profile of mechanical hypersensitivity. Diverse EAE models bearing genetic, clinical and histopathological heterogeneity, show different profiles of sensory and pathological changes and thereby enable studying the mechanistic basis and the diversity of changes in pain perception that are associated with distinct types of MS.


Electromagnetic fields act via activation of voltage-gated calcium channels to produce beneficial or adverse effects.
The direct targets of extremely low and microwave frequency range electromagnetic fields (EMFs) in producing non-thermal effects have not been clearly established. However, studies in the literature, reviewed here, provide substantial support for such direct targets. Twenty-three studies have shown that voltage-gated calcium channels (VGCCs) produce these and other EMF effects, such that the L-type or other VGCC blockers block or greatly lower diverse EMF effects. Furthermore, the voltage-gated properties of these channels may provide biophysically plausible mechanisms for EMF biological effects. Downstream responses of such EMF exposures may be mediated through Ca(2+) /calmodulin stimulation of nitric oxide synthesis. Potentially, physiological/therapeutic responses may be largely as a result of nitric oxide-cGMP-protein kinase G pathway stimulation. A well-studied example of such an apparent therapeutic response, EMF stimulation of bone growth, appears to work along this pathway. However, pathophysiological responses to EMFs may be as a result of nitric oxide-peroxynitrite-oxidative stress pathway of action. A single such well-documented example, EMF induction of DNA single-strand breaks in cells, as measured by alkaline comet assays, is reviewed here. Such single-strand breaks are known to be produced through the action of this pathway. Data on the mechanism of EMF induction of such breaks are limited; what data are available support this proposed mechanism. Other Ca(2+) -mediated regulatory changes, independent of nitric oxide, may also have roles. This article reviews, then, a substantially supported set of targets, VGCCs, whose stimulation produces non-thermal EMF responses by humans/higher animals with downstream effects involving Ca(2+) /calmodulin-dependent nitric oxide increases, which may explain therapeutic and pathophysiological effects.


Candida albicans and Candida parapsilosis induce different T-cell responses in human peripheral blood mononuclear cells.
Candida parapsilosis is the third most frequent cause of candidemia. Despite its clinical importance, little is known about the human immunological response to C. parapsilosis. In this study, we compared the cytokine responses evoked by Candida albicans and C. parapsilosis. C. parapsilosis-stimulated human peripheral blood mononuclear cells (PBMCs) produced similar quantities of tumor necrosis factor α and interleukin 6 and slightly lower amounts of interleukin 1β, compared with C. albicans-stimulated cells. PBMCs stimulated with C. parapsilosis displayed a skewed T-helper cell response, producing more interleukin 10 and less interferon γ than cells stimulated with C. albicans. Notably, C. parapsilosis induced much less interleukin 17 and interleukin 22 production as compared to C. albicans. Inhibition of the 3 classical mitogen-activated protein kinases (p38 kinase, ERK, and JNK) revealed kinase-dependent differences in reductions in cytokine production by the 2 Candida species. Decreased cytokine production after inhibition of dectin 1 revealed that this receptor plays a major role in the recognition of both C. albicans and C. parapsilosis. These data improve understanding of the immune response triggered by C. parapsilosis, a first step for the future design of immunotherapeutic strategies for these infections.


Reproductive behavior and fitness components in male Drosophila melaogaster are non-linearly affected by the number of male co-inhabitants early in adult life.
Although multiple lines of evidence suggest that early adult life is very important in shaping the reproductive behavior of males, few studies have looked at the fitness consequences of the variation in reproductive behavior induced by differences in early life experience of males. Using a long term laboratory adapted population of Drosophila melanogaster Meigen (Diptera: Drosophilidae), early life experience, in terms of co-inhabitant numbers, was found to affect male mating behavior and at least one fitness component. However, in contrast to previous studies, a non-linear relationship was found between early life experience and fitness components and a significant effect of co-inhabitant number on copulation duration and sperm defense. Both these traits showed a sharp increase as the co-inhabitant numbers changed from 1 to 16. However, there was a decline in the trait values as the co-inhabitant number increased further. The probable causes for the observed non-linear pattern of responses are discussed.


Is hand-held dynamometry useful for the measurement of quadriceps strength in older people? A comparison with the gold standard Bodex dynamometry.
The lower limb muscle strength is an important determinant of physical function in older people. However, measurement in clinical and epidemiological settings has been limited because of the requirement for large-scale equipment. A protocol using a novel, versatile hand-held dynamometer (HHD) has been developed to measure the quadriceps strength in a supine position. The objective of this study was to assess the validity of this new methodology for measuring the lower limb muscle strength compared to the gold standard Biodex dynamometer. The supine quadriceps strength was measured twice with each of the Biodex and the HHD in 20 men and women, aged 61-81 years, on their non-dominant leg. The agreement between the peak torques obtained by Biodex and HHD was analyzed. The mean peak Biodex and HHD results were 83.4 +/- (SD) 28.0 Nm and 68.9 +/- 19.6 Nm, respectively. The HHD undermeasured the quadriceps strength by an average of 14.5 Nm (95% CI 8.5, 20.6) compared to the Biodex, and this effect was most marked in the strongest participants. Nevertheless, there was a good correlation between the measures (r = 0.91, p < 0.0001). Classification of individuals into tertiles of muscle strength showed good agreement between the two methods (Kappa = 0.69, p < 0.0001). Our findings suggest that the HHD using a supine positioning offers a feasible, inexpensive, and portable test of quadriceps muscle strength for use in healthy older people. It underestimates the absolute quadriceps strength compared to the Biodex particularly in stronger people, but is a useful tool for ranking muscle strength of older people in epidemiological studies. It may also be of value for quick and objective assessment of physical function in the clinical setting.


The Microbial Rosetta Stone database: A common structure for microbial biosecurity threat agents.
Infectious microorganisms are important to multiple communities engaged in biodefense and biosecurity, including the agencies responsible for health, defense, law enforcement, agriculture, and drug and food safety. Many agencies have created lists of high priority infectious microorganisms to prioritize research efforts or to formally control the possession and distribution of specific organisms or toxins. However, the biological classification of infectious microorganisms is often complex and ambiguous, leading to uncertainty and confusion for scientists involved in biosecurity work. To address this problem, we created a database, known as the Microbial Rosetta Stone, which resolves many of these ambiguities and includes links to additional information on the microbes, such as gene sequence data and scientific literature. Here we discuss the efforts to coordinate organism names from pathogen lists from various governmental agencies according to biological relatedness and show the overlap of high-priority organisms from multiple agencies. To our knowledge, this is the first comprehensive coordination of pathogens, synonyms, and correct taxonomic names. The organized tables and visual aids are freely available at http://www.microbialrosettastone.com. This website provides a single location where access to information on a broad range of disease-causing organisms and toxins is available to members of the biosecurity community.


Cyclin D1 provides a link between development and oncogenesis in the retina and breast.
Mice lacking cyclin D1 have been generated by gene targeting in embryonic stem cells. Cyclin D1-deficient animals develop to term but show reduced body size, reduced viability, and symptoms of neurological impairment. Their retinas display a striking reduction in cell number due to proliferative failure during embryonic development. In situ hybridization studies of normal mouse embryos revealed an extremely high level of cyclin D1 in the retina, suggesting a special dependence of this tissue on cyclin D1. In adult mutant females, the breast epithelial compartment fails to undergo the massive proliferative changes associated with pregnancy despite normal levels of ovarian steroid hormones. Thus, steroid-induced proliferation of mammary epithelium during pregnancy may be driven through cyclin D1.


Population biology of antigen presentation by MHC class I molecules.
In principle, the function of major histocompatibility complex (MHC) molecules is simple: to bind a peptide and engage a T cell. In practice, placing this function within the context of the immune response begs questions of population biology; How does the immune response emerge from the interactions among populations of peptides, T cells and MHC molecules? Within a population of vertebrates, how does MHC polymorphism stamp individuality on the response? Does polymorphism confer differential advantages in responding to parasites? How are the pressures on the MHC reflected in turnover of alleles? The role of mutation, recombination, selection, and drift in the generation and maintenance of MHC class 1 polymorphism are considered.


Oxidative stress induced by UVA photoactivation of the tryptophan UVB photoproduct 6-formylindolo[3,2-b]carbazole (FICZ) inhibits nucleotide excision repair in human cells.
Potentially mutagenic DNA lesions induced by UVB (wavelengths 280-320 nm) are important risk factors for solar ultraviolet (UV) radiation-induced skin cancer. The carcinogenicity of the more abundant UVA (320-400 nm) is less well understood but is generally regarded to reflect its interaction with cellular chromophores that act as photosensitisers. The arylhydrocarbon receptor agonist 6-formylindolo[3,2-b] carbazole (FICZ), is a UVB photoproduct of tryptophan and a powerful UVA chromophore. Combined with UVA, FICZ generates reactive oxygen species (ROS) and induces oxidative DNA damage. Here we demonstrate that ROS generated by FICZ/UVA combinations also cause extensive protein damage in HaCaT human keratinocytes. We show that FICZ/UVA-induced oxidation significantly inhibits the removal of potentially mutagenic UVB-induced DNA photolesions by nucleotide excision repair (NER). DNA repair inhibition is due to FICZ/UVA-induced oxidation damage to the NER proteome and DNA excision repair is impaired in extracts prepared from FICZ/UVA-treated cells. NER protects against skin cancer. As a likely UVB photoproduct of intracellular tryptophan, FICZ represents a de facto endogenous UVA photosensitiser in sun-exposed skin. FICZ formation may increase the risk of solar UV-induced skin cancer by promoting photochemical damage to the NER proteome and thereby preventing the removal of UVB-induced DNA lesions.


Risk Factors for Myocardial Infarction and Stroke Among Sepsis Survivors: A Competing Risks Analysis.
Predictors for post-sepsis myocardial infarction (MI) and stroke are yet to be identified due to the competing risk of death. This study included all hospitalized patients with sepsis from National Health Insurance Research Database of Taiwan between 2000 and 2011. The primary outcome was the first occurrence of MI and stroke requiring hospitalization within 180 days following hospital discharge from the index sepsis episode. The association between predictors and post-sepsis MI and stroke were analyzed using cumulative incidence competing risk model that controlled for the competing risk of death. Among 42 316 patients with sepsis, 1012 (2.4%) patients developed MI and stroke within 180 days of hospital discharge. The leading 5 predictors for post-sepsis MI and stroke are prior cerebrovascular diseases (hazard ratio [HR]: 2.02, 95% confidence interval [CI]: 1.74-2.32), intra-abdominal infection (HR: 1.94, 95% CI: 1.71-2.20), previous MI (HR: 1.81, 95% CI: 1.53-2.15), lower respiratory tract infection (HR: 1.62, 95% CI: 1.43-1.85), and septic encephalopathy (HR: 1.61, 95% CI: 1.26-2.06). Baseline comorbidities and sources of infection were associated with an increased risk of post-sepsis MI and stroke. The identified risk factors may help physicians select a group of patients with sepsis who may benefit from preventive measures, antiplatelet treatment, and other preventive measures for post-sepsis MI and stroke.


Highly efficient one-pot assembly of peptides by double chemoselective coupling.
This study describes a methodological advancement in solution-phase peptide synthesis via the development of a convenient and operational protocol to synthesize oligopeptides in a one-pot three-step cascade method, in which two peptide bonds are introduced chemoselectively. Tri- to hexapeptides were obtained in high global yields (80-95%) with virtually no epimerization as determined via HPLC. The methodology described herein represents a faster, easier and milder approach to the synthesis of peptides, and it operates at equimolar amounts. This protocol comprises the formation of secondary and tertiary amides and is compatible with Z, Boc and Fmoc N-protecting groups as well as the use of d/l and non-proteinogenic amino acids.


Relations of blood inflammatory marker levels with cerebral microbleeds.
Cerebral microbleeds (CMB) are observed in the elderly and have been regarded as one of the manifestations of small vessel disease. Although inflammatory processes have attracted much attention not only in large-artery disease, but also in small vessel disease, their involvement in CMB remains to be determined. The purpose of this study is to clarify relations between inflammatory marker levels and CMB. Four hundred thirty-one patients without histories of cerebrovascular diseases were prospectively enrolled. The presence and number of CMB were assessed on gradient-echo magnetic resonance imaging. As common inflammatory markers, serum levels of high-sensitivity C-reactive protein (hsCRP), interleukin-6 (IL-6), and interleukin-18 (IL-18) were evaluated. CMB were found in 65 patients (15%). In 35 patients, at least one CMB was found in deep locations, but 30 patients had strictly lobar CMB. Levels of hsCRP, IL-6, and IL-18 were higher in patients with CMB than in those without. Logistic regression analyses showed that each 1SD increase in each inflammatory marker level was significantly associated with the presence of CMB after adjustment for age and sex, and after additional adjustment for cardiovascular risk factors, silent lacunar infarction, and white matter hyperintensity. The OR (95% CI) of hsCRP, IL-6, and IL-18 was 1.81 (1.35-2.46), 1.73 (1.18-2.61), and 2.41 (1.44-4.52), respectively. Furthermore, the inflammatory marker levels were associated with both deep and lobar CMB. Higher levels of hsCRP, IL-6, and IL-18 are associated with CMB, in both deep and lobar locations, suggesting the involvement of inflammation in CMB.


A comparative study of pediatric open pyeloplasty, laparoscopy-assisted extracorporeal pyeloplasty, and robot-assisted laparoscopic pyeloplasty.
To compare the outcomes of open pyeloplasty (OP), laparoscopy-assisted extracorporeal (LEXP), and robotic-assisted laparoscopic pyeloplasty (RALP) for ureteropelvic junction obstruction in pediatric patients. We retrospectively reviewed the age-matched cohort of 30 children who underwent OP, 30 who underwent LEXP, and 10 who underwent RALP at a single institution, from 1996 to 2014. Pre- and post-operative variables including success rate were compared among surgical groups. The mean age of the patients was 120.2 months, the Society for Fetal Urology grade was 3.6, the anteroposterior diameter was 3.1 cm, and the renal relative function was 44.0%. The distribution of laterality, mean body mass index, and preoperative anteroposterior pelvic diameter on ultrasound did not differ among groups. The mean length of hospital stay was significantly shorter in the RALP group (3.2 days) than in the OP (6.6 days) and LEXP (5.8 days) groups (p<0.001). The duration of analgesics use was shorter in the RALP group (1.1 days) than in the other groups (p<0.001). During the mean follow-up period of 49.0, 20.1, and 16.6 months, the success rate was 96.7%, 89.7%, and 100% in the OP, LEXP, and RALP groups, respectively, although this difference was not statistically different (p = 0.499). In multivariate regression analysis, the presence of crossing vessels was the only factor that decreased the success rate (hazard ratio: 46.09, 95% confidence interval: 2.41-879.6, p = 0.011). Patients who undergo RALP have a reduced hospital stay and lower use of pain medication; however, there is no difference in the success rates for OP, LP, and RALP surgeries. The presence of crossing vessels is a negative prognostic indicator for surgical outcome regardless of the surgical method.


Symmetry in context: salience of mirror symmetry in natural patterns.
Symmetry is a biologically relevant, mathematically involving, and aesthetically compelling visual phenomenon. Mirror symmetry detection is considered particularly rapid and efficient, based on experiments with random noise. Symmetry detection in natural settings, however, is often accomplished against structured backgrounds. To measure salience of symmetry in diverse contexts, we assembled mirror symmetric patterns from 101 natural textures. Temporal thresholds for detecting the symmetry axis ranged from 28 to 568 ms indicating a wide range of salience (1/Threshold). We built a model for estimating symmetry-energy by connecting pairs of mirror-symmetric filters that simulated cortical receptive fields. The model easily identified the axis of symmetry for all patterns. However, symmetry-energy quantified at this axis correlated weakly with salience. To examine context effects on symmetry detection, we used the same model to estimate approximate symmetry resulting from the underlying texture throughout the image. Magnitudes of approximate symmetry at flanking and orthogonal axes showed strong negative correlations with salience, revealing context interference with symmetry detection. A regression model that included the context-based measures explained the salience results, and revealed why perceptual symmetry can differ from mathematical characterizations. Using natural patterns thus produces new insights into symmetry perception and its possible neural circuits.


Therapeutic efficacy of topical epigallocatechin gallate in murine dry eye.
To study the efficacy of topical epigallocatechin gallate (EGCG) for the treatment of dry eye disease (DED). Seven- to 8-week-old female C57BL/6 mice were housed in the controlled environment chamber to induce DED. Topical 0.01% or 0.1% EGCG, or vehicle, was applied to the eyes of DED mice. Corneal fluorescein staining and the number of corneal CD11b+ cells were assessed in the different groups. Expression of interleukin-1β, tumor necrosis factor-α, chemokine ligand 2, and vascular endothelial growth factor (VEGF)-A/C/D was evaluated by real-time polymerase chain reaction in the corneas at day 9. Corneas were stained for lymphatic vessel endothelial hyaluronan receptor (LYVE)-1 to evaluate lymphangiogenesis, and the terminal transferase dUTP nick end labeling (TUNEL) assay was used to evaluate apoptosis of corneal epithelial cells. Treatment with 0.1% EGCG showed a significant decrease in corneal fluorescein staining compared with the vehicle (24.6%, P = 0.001) and untreated controls (41.9%, P < 0.001). A significant decrease in the number of CD11b+ cells was observed in 0.1% EGCG-treated eyes, compared with the vehicle in the peripheral (23.3%, P = 0.001) and central (26.1%, P = 0.009) corneas. Treatment with 0.1% EGCG was associated with a significant decrease in the corneal expression of interleukin-1β (P = 0.029) and chemokine ligand 2 (P = 0.001) compared with the vehicle and in VEGF-A and VEGF-D levels compared with the untreated group (P = 0.007 and P = 0.048, respectively). EGCG 0.01% also showed a decrease in inflammation at the molecular level but no significant changes in the clinical signs of DED. No cellular toxicity to the corneal epithelium was observed with 0.01% or 0.1% EGCG. Topical EGCG treatment is able to reduce the clinical signs and inflammatory changes in DED by suppressing the inflammatory cytokine expression and infiltration of CD11b+ cells in the cornea.


Atlantoaxial Fusion: Sixteen Years of Epidemiology, Indications, and Complications in New York State.
A retrospective state database analysis. The aim of this study was to describe the epidemiology and complications of as well as indications for primary isolated atlantoaxial fusion. Atlantoaxial fusion involves unique indications, techniques, and complications. There is limited epidemiologic literature focused specifically on this procedure. New York's Statewide Planning and Research Cooperative System database, an all-payer hospitalization reporting system, was queried to identify all patients undergoing primary isolated atlantoaxial fusion in the state from 1997 to 2012. Demographic and clinical data were extracted and analyzed. United States Census Bureau figures were used to calculate population-adjusted surgical rates. One thousand five hundred fifty-nine patients underwent isolated primary atlantoaxial fusion during the study period. The overall population-adjusted annual surgical rate did not change significantly over time. By 2012, individuals aged ≥70 years had the highest incidence of surgery [2.37 per 100,000 population; 95% confidence interval (95% CI) 1.68-3.07]. Medicare was the most common payer (44.0% of claims). Approximately 85% of patients had a Charlson/Deyo Comorbidity Index of zero or one. Over time, a significantly lower proportion of atlantoaxial fusions were attributable to rheumatic disease, and a significantly higher proportion were due to fracture. By 2012, management of fractures was the most common indication for C1-C2 fusion (44.1% of cases). Dysphagia or dysphonia occurred after 0.8% of cases, dural tear after 0.3%, infection after 0.5%, and seroma, hematoma, or hemorrhage after 0.5%. In-hospital mortality was 2.7%, of which 76% had fracture as the surgical indication. Isolated atlantoaxial fusions have been performed at a stable, low level over the past 16 years in New York. Although most of these patients are relatively healthy pre-operatively, approximately one in 10 experience an in-hospital complication and nearly 3% die in-hospital. Knowledge of these risks will hopefully spur further efforts to minimize them and allow for more accurate counseling of patients and their families. 4.


Ser/Thr protein kinase Bβ-NADPH oxidase 2 signaling in thromboinflammation.
Interactions between neutrophils and platelets contribute to the progression of thromboinflammatory disease. However, the regulatory mechanism governing these interactions is poorly understood. The present review focuses on the crucial role of Ser/Thr protein kinase B (AKT)β-NADPH oxidase 2 (NOX2) signaling in regulating neutrophil and platelet activation and their heterotypic interactions under thromboinflammatory conditions. Growing evidence has shown that platelets, leukocytes, and blood coagulation need to be considered to treat thromboinflammatory disease in which inflammation and thrombosis occur concurrently. In addition to plasma proteins and intracellular signaling molecules, extracellular reactive oxygen species (ROS) produced from activated leukocytes could be an important factor in the pathophysiology of thromboinflammatory disease. Recent studies reveal that AKT2-NOX2 signaling has critical roles in Ca mobilization, ROS generation, degranulation, and control of the ligand-binding function of cell surface molecules, thereby promoting heterotypic cell-cell interactions in thromboinflammation. These findings have provided novel insights into attractive therapeutic targets for the prevention and treatment of thromboinflammatory disease. Recent discoveries concerning molecular mechanisms regulating neutrophil-platelet interactions have bridged some gaps in our knowledge of the complicated signaling pathways exacerbating thromboinflammatory conditions.


Micro(glial)-managing executive function: white matter inflammation drives catatonia.
White matter abnormalities are prevalent in neuropsychiatric disorders such as schizophrenia, but it is unclear whether these abnormalities represent a cause or consequence of these disorders. Reduced levels of the myelin protein 2'-3'-cyclic nucleotide 3'-phosphodiesterase (CNP) are associated with the schizophrenic symptom catatonia in both humans and mouse models. In this issue of the JCI, Janova et al. show that reduced CNP levels correlate with catatonia and white matter inflammation in human subjects. Furthermore, they demonstrate that microglial ablation prevents and alleviates catatonic signs in Cnp-/- mice, indicating that microglial-mediated inflammation causes catatonia. Together, this study identifies a cellular mechanism by which subtle myelin abnormalities cause low-grade neuroinflammation and catatonic behavior.


Post-synthetic modulation of the charge distribution in a metal-organic framework for optimal binding of carbon dioxide and sulfur dioxide.
Modulation of pore environment is an effective strategy to optimize guest binding in porous materials. We report the post-synthetic modification of the charge distribution in a charged metal-organic framework, MFM-305-CH3, [Al(OH)(L)]Cl, [(H2L)Cl = 3,5-dicarboxy-1-methylpyridinium chloride] and its effect on guest binding. MFM-305-CH3 shows a distribution of cationic (methylpyridinium) and anionic (chloride) centers and can be modified to release free pyridyl N-centres by thermal demethylation of the 1-methylpyridinium moiety to give the neutral isostructural MFM-305. This leads simultaneously to enhanced adsorption capacities and selectivities (two parameters that often change in opposite directions) for CO2 and SO2 in MFM-305. The host-guest binding has been comprehensively investigated by in situ synchrotron X-ray and neutron powder diffraction, inelastic neutron scattering, synchrotron infrared and 2H NMR spectroscopy and theoretical modelling to reveal the binding domains of CO2 and SO2 in these materials. CO2 and SO2 binding in MFM-305-CH3 is shown to occur via hydrogen bonding to the methyl and aromatic-CH groups, with a long range interaction to chloride for CO2. In MFM-305 the hydroxyl, pyridyl and aromatic C-H groups bind CO2 and SO2 more effectively via hydrogen bonds and dipole interactions. Post-synthetic modification via dealkylation of the as-synthesised metal-organic framework is a powerful route to the synthesis of materials incorporating active polar groups that cannot be prepared directly.


An environmental justice perspective on ecosystem services.
Mainstreaming of ecosystem service approaches has been proposed as one path toward sustainable development. Meanwhile, critics of ecosystem services question if the approach can account for the multiple values of ecosystems to diverse groups of people, or for aspects of inter- and intra-generational justice. In particular, an ecosystem service approach often overlooks power dimensions and capabilities that are core to environmental justice. This article addresses the need for greater guidance on incorporating justice into ecosystem services research and practice. We point to the importance of deep engagement with stakeholders and rights holders to disentangle contextual factors that moderate justice outcomes on ecosystem service attribution and appropriation in socio-political interventions. Such a holistic perspective enables the integration of values and knowledge plurality for enhancing justice in ecosystem services research. This broadened perspective paves a way for transformative ecosystem service assessments, management, and research, which can help inform and design governance structures that nourish human agency to sustainably identify, manage, and enjoy ecosystem services for human wellbeing.


Experimental observation of nonlinear pulse compression in nonuniform Bragg gratings.
We demonstrate a scheme for optical pulse compression by cross-phase modulation that utilizes a nonuniform Bragg grating to work in reflection. Our scheme is similar to the conventional optical pushbroom, which works in transmission. This reflection geometry has the advantage of allowing the compressed signal to be observed easily, as it is spatially separate from the pump. This is to our knowledge the first nonlinear effect to be observed that requires a nonuniform grating.


Activation of the erythropoietin receptor by the Friend spleen focus-forming virus gp55 glycoprotein induces constitutive protein tyrosine phosphorylation.
The erythropoietin receptor (EPO-R) can be activated to signal cell growth by binding either EPO or gp55, the Friend spleen focus-forming virus (SFFV) glycoprotein. EPO binding induces tyrosine kinase activity and rapid tyrosine phosphorylation of several cellular substrates. To test for gp55-induced tyrosine kinase activity, we performed immunoblots on two murine cell lines that stably express EPO-R and gp55. Stimulation of the parental cell line, Ba/F3, with murine interleukin-3 (IL-3) resulted in rapid, dose-dependent tyrosine phosphorylation of a 97-Kd substrate. Stimulation with IL-3 or EPO of the Ba/F3 cells expressing the recombinant EPO-R (Ba/F3-EPO-R) resulted in tyrosine phosphorylation of the same p97 substrate. These latter cells, when transformed to growth factor-independence by the Friend gp55 glycoprotein, exhibited constitutive tyrosine phosphorylation of the 97-Kd substrate. Other growth factor-independent Ba/F3 subclones, transformed with either the oncoprotein, v-abl, or with a constitutively activated EPO-R, also had constitutive phosphorylation of a 97-Kd substrate. In CTLL-2-EPO-R cells, a T-lymphocyte line stably transfected with the EPO-R, the 97-Kd substrate was tyrosine-phosphorylated in response to IL-2 or EPO. The 97-Kd protein was constitutively phosphorylated in CTLL-2-EPO-R-gp55 cells. In conclusion, a 97-Kd protein found in two murine cell lines is tyrosine-phosphorylated in response to multiple growth factors and viral oncoproteins, and appears to be a central phosphoprotein in signal transduction.


Rationale and design of a trial to personalize risk assessment in familial coronary artery disease.
The lifetime risk of coronary artery disease (CAD) is doubled in people with a family history of premature disease, yet this risk is not captured in most 5- or 10-year risk assessment algorithms. Coronary artery calcium scoring (CCS) is a marker of subclinical CAD risk, which has been shown in observational studies to provide prognostic information that is incremental to clinical assessment; is relatively inexpensive; and is performed with a small radiation dose. However, the use of CCS in guiding prevention is not strongly supported by guidelines. Showing definitive evidence of the efficacy and cost-effectiveness of CCS is therefore of importance. The proposed randomized controlled trial of the use of CCS will be targeted to 40- to 70-year-old first-degree relatives of patients with CAD onset <60 years old or second-degree relatives of patients with onset <50 years old. Control patients will undergo standard risk scoring and be blinded to CCS results. In the intervention group, primary prevention in patients undergoing CCS will be informed by this score. At 3 years, effectiveness will be assessed on change in plaque volume at computed tomography coronary angiography, the extent of which has been strongly linked to outcome. The CAUGHT-CAD trial will provide evidence to inform the guidelines regarding the place of CCS in decision making regarding primary prevention of patients with a family history of premature disease.


High-fidelity detection and sorting of nanoscale vesicles in viral disease and cancer.
Biological nanoparticles, including viruses and extracellular vesicles (EVs), are of interest to many fields of medicine as biomarkers and mediators of or treatments for disease. However, exosomes and small viruses fall below the detection limits of conventional flow cytometers due to the overlap of particle-associated scattered light signals with the detection of background instrument noise from diffusely scattered light. To identify, sort, and study distinct subsets of EVs and other nanoparticles, as individual particles, we developed nanoscale Fluorescence Analysis and Cytometric Sorting (nanoFACS) methods to maximise information and material that can be obtained with high speed, high resolution flow cytometers. This nanoFACS method requires analysis of the instrument background noise (herein defined as the "reference noise"). With these methods, we demonstrate detection of tumour cell-derived EVs with specific tumour antigens using both fluorescence and scattered light parameters. We further validated the performance of nanoFACS by sorting two distinct HIV strains to >95% purity and confirmed the viability (infectivity) and molecular specificity (specific cell tropism) of biological nanomaterials sorted with nanoFACS. This nanoFACS method provides a unique way to analyse and sort functional EV- and viral-subsets with preservation of vesicular structure, surface protein specificity and RNA cargo activity.


Ventilator-associated pneumonia in patients with adult respiratory distress syndrome: CT evaluation.
To determine the diagnostic accuracy of computed tomography (CT) for pneumonia in patients with adult respiratory distress syndrome (ARDS). CT scans were obtained within 1 week of bronchoscopic sampling in 31 patients receiving mechanical ventilation for ARDS for more than 48 hours. Of 11 patients with pneumonia, five developed symptoms less than 11 days after the onset of ARDS (early ARDS). CT scans were rated for pneumonia independently by four radiologists who were unaware of the clinical diagnosis. Diagnostic accuracy was defined by means of the area under the receiver operating characteristic curve, or A2. Diagnostic accuracy for pneumonia was fair (A2 = 0.69 +/- 0.04 [standard error]) owing to 70% true-negative ratings (vs 59% true-positive ratings). The generalizability coefficient was good (0.79). No single CT finding was significantly different for the presence of pneumonia. Nondependent opacities predominated in 10 (91%) of 11 patients with pneumonia and 12 (60%) of 20 without pneumonia. Nondependent opacities predominated in nine (56%) of 16 patients with early ARDS and 13 (87%) of 15 with late ARDS. CT has fair diagnostic accuracy for ventilator-associated pneumonia in patients with ARDS owing primarily to identification of patients without pneumonia. No single CT sign was significantly different for pneumonia, but dependent atelectasis was more common in patients with early ARDS without pneumonia.


Nitric oxide limits coronary vasoconstriction by a shear stress-dependent mechanism.
Increases in shear stress promote coronary vasodilation by stimulating the production of nitric oxide (NO). Whether shear stress-induced NO production also limits vasoconstriction in the coronary microcirculation in vivo is unknown. Accordingly, we measured microvascular diameter and flow velocity in the beating heart along with estimated blood viscosity to calculate shear stress during vasoconstriction with endothelin or vasopressin. Measurements were repeated in the presence of NG-monomethyl-L-arginine (L-NMMA) to inhibit NO production and BQ-788 to block NO-linked endothelin type B receptors. BQ-788 did not augment steady-state constriction to endothelin, suggesting that NO production via activation of this receptor is inconsequential. L-NMMA potentiated constriction to both agonists, particularly in small arteries (inner diameter >120 microm). Shear stresses in small arteries were elevated during constriction and further elevated during constriction after L-NMMA. These observations suggest that NO production limits vasoconstriction in the coronary microcirculation and that the principal stimulus for this governance is elevated shear stress. The degree of shear stress moderation of constriction is heterogeneously distributed, with small arteries displaying a higher degree of shear stress regulation than arterioles. These results provide the strongest evidence to date that shear stress-mediated production of NO exerts a "braking" influence on constriction in the coronary microcirculation.


Combination antiangiogenesis therapy with marimastat, captopril and fragmin in patients with advanced cancer.
Marimastat, low molecular weight heparins and captopril have antiangiogenic activity in vitro and in animal models. We studied the safety and efficacy of the combination of these drugs in patients with advanced cancer. In all, 50 patients were enrolled. Captopril was given orally at a dose of 50 mg bd daily. Fragmin was administered as a daily subcutaneous injection of 200 units kg(-1) for the first 28 days and 5000 units thereafter. Marimastat was given at 10 mg bd orally. Serum, plasma and urinary angiogenic factors were measured at baseline and after 1 month of treatment. Inhibition of release of tumour necrosis factor alpha (TNF-alpha) from peripheral lymphocytes was used as a surrogate pharmacodynamic end point. There was one case of haemorrhagic stroke and one upper gastrointestinal haemorrhage. The commonest toxicity was myalgia. One of 10 patients with renal cancer had a partial response, and three patients had a prolonged period of stable disease. The treatment significantly inhibited phytohaemagglutinin (PHA)-stimulated TNF-alpha release from patient's lymphocytes. The combination of marimastat, fragmin and captopril is well tolerated and has in vivo activity. Inhibition of PHA-stimulated TNF-alpha release from lymphocytes is a surrogate pharmacodynamic marker of metalloprotease inhibition.


Improving adherence to screening follow-up among women with abnormal Pap smears: results from a large clinic-based trial of three intervention strategies.
In a large randomized trial involving over 2,000 women with abnormal cervical cytology (pap smear), three clinic-based interventions were tested as strategies to increase return rates for screening follow-up: 1) a personalized follow-up letter and pamphlet; 2) a slide-tape program on pap smears; and 3) transportation incentives (bus passes/parking permits). The three interventions were evaluated using a 2 x 2 x 2 factorial design. Results of this study confirm a high rate of loss to screening follow-up (i.e., no return visits) among women with abnormal pap smears (29% overall), with substantial variability among the 12 participating clinics (13% to 42/%). For the sample as a whole, both transportation incentives and the combined intervention condition of personalized follow-up and slide-tape program had a significant positive impact on screening follow-up. However, transportation incentives emerged as the dominant intervention condition among patient subgroups that can be characterized as more disadvantaged socioeconomically and at higher risk of developing cervical cancer, including patients receiving care from the county health department (odds ratio (OR) = 1.51; P less than .05); patients without health insurance (OR = 1.77; P less than .01); and patients with more severe pap smear results (OR = 1.71; P less than .05). In contrast, among patient subgroups that can be characterized as relatively more advantaged and at lower risk of developing cervical cancer, only the combined intervention condition of personalized follow-up and slide-tape program was associated with a higher patient return rate. Subgroups reflecting this pattern included patients seen in noncounty clinics (OR = 4.54; P less than .05) and patients with less severe pap smear results (OR = 5.16; P less than .01). The implications of these findings are discussed in terms of designing clinic-based interventions to improve screening follow-up.


Human pregnane X receptor antagonists and agonists define molecular requirements for different binding sites.
The pregnane X receptor (PXR) is an important transcriptional regulator of the expression of xenobiotic metabolism and transporter genes. The receptor is promiscuous, binding many structural classes of molecules that act as agonists at the ligand-binding domain, triggering up-regulation of genes, increasing the metabolism and excretion of therapeutic agents, and causing drug-drug interactions. It has been suggested that human PXR antagonists represent a means to counteract such interactions. Several azoles have been hypothesized to bind the activation function-2 (AF-2) surface on the exterior of PXR when agonists are concurrently bound in the ligand-binding domain. In the present study, we have derived novel computational models for PXR agonists using different series of imidazoles, steroids, and a set of diverse molecules with experimental PXR agonist binding data. We have additionally defined a novel pharmacophore for the steroidal agonist site. All agonist pharmacophores showed that hydrophobic features are predominant. In contrast, a qualitative comparison with the corresponding PXR antagonist pharmacophore models using azoles and biphenyls showed that they are smaller and hydrophobic with increased emphasis on hydrogen bonding features. Azole antagonists were docked into a proposed hydrophobic binding pocket on the outer surface at the AF-2 site and fitted comfortably, making interactions with key amino acids involved in charge clamping. Combining computational and experimental data for different classes of molecules provided strong evidence for agonists and antagonists binding distinct regions on PXR. These observations bear significant implications for future discovery of molecules that are more selective and potent antagonists.


REST is a crucial regulator for acquiring EMT-like and stemness phenotypes in hormone-refractory prostate cancer.
Castration-resistance prostate cancer (CRPC), also known as hormone-refractory prostate cancer (HRPC), requires immediate attention since it is not only resistant to androgen ablation, chemo- and radiotherapy, but also highly metastatic. Increasing evidence suggests that enrichment of neuroendocrine (NE) cells is associated with CRPC. Here, combined RNA-seq and ChIP-seq analysis reveals that REST is involved in epithelial-mesenchymal transition (EMT) and stemness acquisition in NE differentiated prostate cancer (PCa) cells via direct transcriptional repression of Twist1 and CD44. Specifically we show that short-term knockdown of REST induces NE differentiation of LNCaP cells. Long-term REST knockdown enhanced the expression of Twist1 and CD44, cell migration and sphere formation. Overexpression of REST in hormone-refractory CWR22Rv1 PCa cells significantly reduces Twist1 and CD44 expression, cell migration and sphere formation. Collectively, our study uncovers REST in regulating EMT and stemness properties of NE PCa cells and suggests that REST is a potential therapeutic target for CRPC.


Place conditioning: age-related changes in the rewarding and aversive effects of alcohol.
Alcohol abuse levels are very high in adolescents, creating a significant societal issue. It has been shown that people who begin alcohol use as adolescents are more likely to become addicts than people who initiate alcohol use as adults. It is important to note that the development of addiction in humans is more rapid with initiation in adolescence than in adulthood. To determine changes in the reinforcing efficacy of alcohol as a function of adolescent development, we used a place-conditioning paradigm. In this study, we assessed the ability of ethanol to support a conditioned place preference (CPP) or aversion. Animals [postnatal days (PND) 25, 35, 45, and 60] were tested for alcohol-induced conditioning in response to a range of ethanol doses (0.2, 0.5, 1.0, and 2.0 g/kg intraperitoneally) or saline. In general, there was a trend for alcohol to produce an aversion to the ethanol-paired compartment at higher doses. These patterns differed significantly as a function of age. Younger animals (PND 25) exhibited a CPP to a low dose and an aversion at high doses. Late-adolescent (PND 45) animals exhibited a CPP at two moderate doses but a conditioned place aversion at the highest dose. PND 35 and 60 animals did not exhibit a CPP at any examined dose, and PND 60 animals exhibited a progressive aversion with increasing dose. The data show that the developmental processes of adolescence influence general responsiveness to alcohol. Specifically, late-adolescent animals (PND 45) seem to prefer doses of alcohol that are either not reinforcing (0.5 g/kg) or are aversive (1.0 g/kg) at other ages. These processes need to be examined thoroughly to understand the development of addiction in adolescence. This is especially important given that alcohol abuse in adolescence may interfere with the usual pattern of brain development as it relates to alcohol reinforcement.


Short report: prevalence of hepatitis C viral antibody among Brazilian children, adolescents, and street youths.
The prevalence of hepatitis C antibodies (anti-HCV) was investigated in 1,378 central Brazilian children, students, and street youths (homeless adolescents without family links or adolescents working in poorly paid activities). Sera were tested with a second generation enzyme-linked immunosorbent assay, and positive samples were retested by a confirmatory assay (line immunoassay). All children attending day care centers were anti-HCV negative. Only one (0.2%) adolescent was positive in the student group. However, higher positivity was found in street youths; four (1.0%) living at home and three (3.0%) living in the streets and anti-HCV antibodies. Among these, the prevalence of anti-HCV increased significantly with age from 0% in the 9-12-year-old group to 6.9% in the 17-20-year-old group. Risk factors including blood transfusion, tattooing, intravenous drug use, and sexual intercourse with multiple partners were significantly associated with the presence of anti-HCV in street youths. These results indicate that apparently healthy children and adolescents attending day care centers or primary schools in central Brazil have a low exposure to HCV infection, but street youths in the same area are at risk for infection with this virus.