Relationship of Vascular Endothelial Growth Factor C, a Lymphangiogenesis Modulator, With Edema Formation, Congestion and Outcomes in Acute Heart Failure.
Although vascular endothelial growth factor C (VEGF-C) is a known lymphangiogenesis modulator, its relationship with congestion formation and outcomes in acute heart failure (AHF) is unknown. Serum VEGF-C levels were measured in 237 patients hospitalized for AHF. The population was stratified by VEGF-C levels and linked with clinical signs of congestion and outcomes. The study's population was divided in VEGF-C tertiles: low (median [Q25-Q75]: 33 [15-175]), medium (606 [468-741]) and high (1141 [968-1442] pg/mL). The group with low VEGF-C on admission presented with the highest prevalence of severe lower-extremity edema (low VEGF-C vs medium VEGF-C vs high VEGF-C): 30% vs 13% vs 20%; P = 0.02); the highest percentage of patients with ascites: 22% vs 9% vs 6%; P = 0.006; and the lowest proportion of patients with pulmonary congestion: 22% vs 30% vs 46%; P = 0.004. The 1-year mortality rate was the highest in the low VEGF-C tertile: 35% vs 28% vs 18%, respectively; P = 0.049. The same pattern was observed for the composite endpoint (death and AHF rehospitalization): 45% vs 43% vs 26%; P = 0.029. The risks of death at 1-year follow-up and composite endpoint were significantly lower in the high VEGF-C group. Low VEGF-C was associated with more severe signs of congestion (signs of fluid accumulation) and adverse clinical outcomes.


Modulation of integrin-linked kinase nucleo-cytoplasmic shuttling by ILKAP and CRM1.
Integrin-linked kinase (ILK) plays key roles in a variety of cell functions, including cell proliferation, adhesion and migration. Within the cell, ILK localizes to multiple sites, including the cytoplasm, focal adhesion complexes that mediate cell adhesion to extracellular substrates, as well as cell-cell junctions in epidermal keratinocytes. Central to understanding ILK function is the elucidation of the mechanisms that regulate its subcellular localization. We now demonstrate that ILK is imported into the nucleus through sequences in its N-terminus, via active transport mechanisms that involve nuclear pore complexes. In addition, nuclear ILK can be rapidly exported into the cytoplasm through a CRM1-dependent pathway, and its export is enhanced by the type 2C protein phosphatase ILKAP. Nuclear localization of ILK in epidermal keratinocytes is associated with increased DNA synthesis, which is sensitive to inhibition by ILKAP. Our studies demonstrate the importance for keratinocyte proliferation of ILK regulation through changes in its subcellular localization, and establish ILKAP and CRM1 as pivotal modulators of ILK subcellular distribution and activity in these cells.


Vitreous hemorrhage and Rhegmatogenous retinal detachment that developed after botulinum toxin injection to the extraocular muscle: case report.
The authors report a case of a rare complication that occurred after botulinum toxin injection to the extraocular muscle, which was easily overlooked and successfully corrected by surgery. A 34-year-old female patient visited our clinic for diplopia and ocular motility disorder after removal of an epidermoid tumor of the brain. At her initial visit, her best-corrected visual acuity (BCVA) was 20/20 for both eyes. An alternate cover test showed 45 prism-diopter esotropia and 3 prism-diopter hypertropia in the right eye. Following 6 months of observation, the deviation of the strabismus did not improve, and botulinum toxin was injected into the right medial rectus (RMR). After 6 days, she visited our clinic with decreased visual acuity of her right eye. The BCVA was found to be 20/50 for her right eye. Funduscopic examination presented a retinal tear inferonasal to the optic disc with preretinal hemorrhage. Subretinal fluid nasal to the fovea was seen on optical coherence tomography (OCT). Barrier laser photocoagulation was done around the retinal tear; however, her visual acuity continued to decrease, and vitreous hemorrhage and subretinal fluid at the lesion did not improve. In addition, a newly developed epiretinal membrane was seen on OCT. An alternate cover test presented 30 prism-diopter right esotropia. 19 weeks after RMR botulinum toxin injection, she received pars plana vitrectomy, membranectomy, endolaser barrier photocoagulation, and intravitreal bevacizumab (Avastin®) injection. After 4 months, her visual acuity improved to 20/20, and only 4 prism-diopter of right hypertropia and 3 prism-diopter of exotropia were noted. Vitreous opacity and the epiretinal membrane were completely removed, as confirmed by funduscopic and examination. Sudden loss of vision after injection of botulinum toxin into the extraocular muscle may suggest a serious complication, and a prompt, thorough ophthalmic examination should be performed. If improvements are not observed, rapid surgical intervention is recommended to prevent additional complications.


Interactions between suspension characteristics and physicochemical properties of silver and copper oxide nanoparticles: a case study for optimizing nanoparticle stock suspensions using a central composite design.
The preparation of a stable nanoparticle stock suspension is the first step in nanotoxicological studies, but how different preparation methods influence the physicochemical properties of nanoparticles in a solution, even in Milli-Q water, is often under-appreciated. In this study, a systematic approach using a central composite design (CCD) was employed to investigate the effects of sonication time and suspension concentration on the physicochemical properties (i.e. hydrodynamic diameter, zeta potential and ion dissolution) of silver (Ag) and copper oxide (CuO) nanoparticles (NPs) and to identify optimal conditions for suspension preparation in Milli-Q water; defined as giving the smallest particle sizes, highest suspension stability and lowest ion dissolution. Indeed, all the physicochemical properties of AgNPs and CuONPs varied dramatically depending on how the stock suspensions were prepared and differed profoundly between nanoparticle types, indicating the importance of suspension preparation. Moreover, the physicochemical properties of AgNPs and CuONPs, at least in simple media (Milli-Q water), behaved in predictable ways as a function of sonication time and suspension concentration, confirming the validity of our models. Overall, the approach allows systematic assessment of the influence of various factors on key properties of nanoparticle suspensions, which will facilitate optimization of the preparation of nanoparticle stock suspensions and improve the reproducibility of nanotoxicological results. We recommend that further attention be given to details of stock suspension preparation before conducting nanotoxicological studies as these can have an important influence on the behavior and subsequent toxicity of nanoparticles.


Ultra-hypofractionated versus conventionally fractionated radiotherapy for prostate cancer (HYPO-RT-PC): patient-reported quality-of-life outcomes of a randomised, controlled, non-inferiority, phase 3 trial.
The HYPO-RT-PC trial compared conventionally fractionated radiotherapy with ultra-hypofractionated radiotherapy in patients with localised prostate cancer. Ultra-hypofractionation was non-inferior to conventional fractionation regarding 5-year failure-free survival and toxicity. We aimed to assess whether patient-reported quality of life (QOL) differs between conventional fractionation and ultra-hypofractionation up to 6 years after treatment in the HYPO-RT-PC trial. HYPO-RT-PC is a multicentre, open-label, randomised, controlled, non-inferiority, phase 3 trial done in 12 centres (seven university hospitals and five county hospitals) in Sweden and Denmark. Inclusion criteria were histologically verified intermediate-to-high-risk prostate cancer (defined as T1c-T3a with one or two of the following risk factors: stage T3a; Gleason score ≥7; and prostate-specific antigen 10-20 ng/mL with no evidence of lymph node involvement or distant metastases), age up to 75 years, and WHO performance status 0-2. Participants were randomly assigned (1:1) to conventional fractionation (78·0 Gy in 39 fractions, 5 days per week for 8 weeks) or ultra-hypofractionation (42·7 Gy in seven fractions, 3 days per week for 2·5 weeks) via a minimisation algorithm with stratification by trial centre, T-stage, Gleason score, and prostate-specific antigen. QOL was measured using the validated Prostate Cancer Symptom Scale (PCSS) and European Organization for Research and Treatment of Cancer Quality-of-Life Questionnaire (EORTC QLQ-C30) at baseline, the end of radiotherapy, months 3, 6, 12, and 24 after radiotherapy, every other year thereafter up to 10 years, and at 15 years. The primary endpoint (failure-free survival) has been reported elsewhere. Here we report QOL, a secondary endpoint analysed in the per-protocol population, up to 6 years after radiotherapy. The HYPO-RT-PC trial is registered with the ISRCTN registry, ISRCTN45905321. Between July 1, 2005, and Nov 4, 2015, 1200 patients were enrolled and 1180 were randomly assigned (conventional fractionation n=591, ultra-hypofractionation n=589); 1165 patients (conventional fractionation n=582, ultra-hypofractionation n=583) were included in this QOL analysis. 158 (71%) of 223 patients in the conventional fractionation group and 146 (66%) of 220 in the ultra-hypofractionation group completed questionnaires at 6 years. The median follow-up was 48 months (IQR 25-72). In seven of ten bowel symptoms or problems the proportion of patients with clinically relevant deteriorations at the end of radiotherapy was significantly higher in the ultra-hypofractionation group than in the conventional fractionation group (stool frequency [p<0·0001], rush to toilet [p=0·0013], flatulence [p=0·0013], bowel cramp [p<0·0001], mucus [p=0·0014], blood in stool [p<0·0001], and limitation in daily activity [p=0·0014]). There were no statistically significant differences in the proportions of patients with clinically relevant acute urinary symptoms or problems (total 14 items) and sexual functioning between the two treatment groups at end of radiotherapy. Thereafter, there were no clinically relevant differences in urinary, bowel, or sexual functioning between the groups. At the 6-year follow-up there was no difference in the incidence of clinically relevant deterioration between the groups for overall urinary bother (43 [33%] of 132 for conventional fractionation vs 33 [28%] of 120 for ultra-hypofractionation; mean difference 5·1% [95% CI -4·4 to 14·6]; p=0·38), overall bowel bother (43 [33%] of 129 vs 34 [28%] of 123; 5·7% [-3·8 to 15·2]; p=0·33), overall sexual bother (75 [60%] of 126 vs 59 [50%] of 117; 9·1% [-1·4 to 19·6]; p=0·15), or global health/QOL (56 [42%] of 134 vs 46 [37%] of 125; 5·0% [-5·0 to 15·0]; p=0·41). Although acute toxicity was higher for ultra-hypofractionation than conventional fractionation, this long-term patient-reported QOL analysis shows that ultra-hypofractionation was as well tolerated as conventional fractionation up to 6 years after completion of treatment. These findings support the use of ultra-hypofractionation radiotherapy for intermediate-to-high-risk prostate cancer. The Nordic Cancer Union, the Swedish Cancer Society, and the Swedish Research Council.


High-power sampled-grating-based master oscillator power amplifier system with 23.5 nm wavelength tuning around 970 nm.
Tunable high-power diode lasers are key components in various established and emerging applications. In this work, we present a compact hybrid master oscillator power amplifier (MOPA) laser system. The system utilizes a tunable GaAs-based sampled-grating (SG) distributed Bragg reflector (DBR) laser as the master oscillator (MO), which emits around a wavelength of 970 nm in a single longitudinal mode with a spectral width below 20 pm. The SG-DBR laser consists of two SGs, each of which can be thermally tuned with microheaters. By tuning one of the two SGs, a discrete wavelength tuning of 21.1 nm can be obtained with a Vernier mode spacing of about 2.3 nm. By tuning both SGs, 23.5 nm of quasi-continuous tuning is obtained, with a mode spacing of about 115 pm. The coupling of the beam emitted by the MO into a tapered power amplifier provides an amplified output power in the watt range having a nearly diffraction-limited beam with a propagation factor of M1/e22=1.6. The combination of high power and wide wavelength tuning in a compact system makes this light source ideal for, among other things, nonlinear frequency conversion.


[The regulation system of brain aromatase activity; effects of androgens on hypothalamic aromatase in the male rat].
Recently, some studies have found the greatest aromatase activity in brain areas associated with sexual differention and sexual behavior, namely the hypothalamic and limbic structures. We studied the regulation of aromatase activity in the hypothalamic area of male rats, using a sensitive in vitro assay which measures the amount of 3H2O formed by tissue homogenates during the conversion of [1 beta-3H] androstenedione to estrogen. After castration, hypothalamic aromatase activity was significantly decreased (P less than 0.01), and seminal vesicle (SV) and prostate (PR) weights were also significantly decreased (P less than 0.01). Castrated male rats were given testosterone (T), 5 alpha-dihydrotestosterone (DHT), 5 alpha-androstane-3 alpha, 17 beta-diol (A3 alpha), 5 alpha-androstane-3 beta and 17 beta-diol(A3 beta) in various doses (200-1000 micrograms/day) for 10 days, and were given 600 micrograms/day T, DHT, A3 alpha and A3 beta for various durations (1-10 days). We found that T, DHT and A3 alpha but not A3 beta reversed the effects of castration on the hypothalamic aromatase activity. The order of this reversible effect of androgens was as follows: T greater than or equal to DHT greater than A3 alpha. T, DHT, A3 alpha and A3 beta increased SV and PR weights, and the order of this effect was as follows: DHT greater than T greater than A3 alpha much greater than A3 beta. We administered the antiandrogen (flutamide) to intact male rats (8 mg/day for 6 days). Flutamide decreased hypothalamic aromatase activity at the same level as that of castrated rats. Likewise, administration of both flutamide and T to castrated rats blocked the T-induced increase in hypothalamic aromatase activity and accessory sexual organ weight. From these results, we suggest that T, DHT and A3 alpha regulated hypothalamic aromatase activity, that T was the most effective of the androgens, and that was different from peripheral androgen target organs.


Psychosocial correlates of safe sex self-efficacy among in-school adolescent girls in Lagos, Nigeria.
Adolescent girls in Nigeria are at heightened risk for HIV and other sexually transmitted infections. However, there are limited studies on psychosocial factors that are associated with safe sex intentions among this population. Self-efficacy has been established as an important correlate of behavioral intentions and the actual behavior. The objective of this research was to examine how key psychosocial factors such as social support, parental monitoring, and future orientation influence perceived safe sex self-efficacy among in-school adolescent girls in Nigeria. Furthermore, we assessed the associations between these psychosocial factors and HIV-related knowledge and safe sex self-efficacy. A self-administered questionnaire was distributed to 426 adolescent girls attending public and private school systems in Lagos, Nigeria. Multiple linear regression was used to evaluate the influence of psychosocial and demographic factors on safe sex self-efficacy. Further, stratified analysis was conducted to compare the estimates between participants attending public schools (n = 272) and those attending private schools (n = 154). Results from the study show that future orientation (β = 0.17; p < 0.05), participants age (β = 0.14; p < 0.05), and HIV knowledge accuracy (β = 0.17; p < 0.05) were associated with safe sex self-efficacy. Future orientation remained statistically significant in the sub-group analysis among participants attending public (β = 0.13; p < 0.05) and private schools (β = 0.24; p < 0.05). Among participants attending public schools, HIV accuracy (β = 0.2; p < 0.05) remained a significant correlate of safe sex self-efficacy while this association dissipated among private school attendees. These findings point to the importance of including future orientation strategies in interventions developed for in-school adolescent girls in Nigeria. School-based interventions that increase positive future orientation outcomes may be beneficial to improve safe sex intentions among adolescent girls in Nigeria.


Characterization of two recombination-complementation groups of Uukuniemi virus temperature-sensitive mutants.
With the aim of isolating temperature-sensitive (ts) mutants defective in virus maturation or glycoprotein transport, Uukuniemi virus, a bunyavirus, was mutagenized with N-methyl-N'-nitro-N-nitrosoguanidine. Out of 13 initial clones unable to grow at 39 degrees C (non-permissive temperature), five mutants which grew to titres above 10(7) p.f.u./ml at 32 degrees C (permissive temperature) were selected for further studies. The mutants fell into two coinciding recombination-complementation groups. Three group I mutants ( ts7 , 8 and 12) and two group II mutants ( ts6 and 11) synthesized all three RNA segments and were able to form the corresponding nucleoproteins at 39 degrees C. Thus, members of these two recombination groups had a RNA-positive phenotype. All five mutants showed immunofluorescence when cells were stained at 39 degrees C using a double-staining technique employing monoclonal antibodies against the glycoproteins G1 or G2, and polyclonal antibodies against the nucleoprotein, N. We have previously shown that in cells infected with wild-type virus both the G1/G2 and the N proteins accumulate in the Golgi complex, the site of virus maturation. In cells infected with ts12 , accumulation of G1 and G2, but not N protein, was observed in the Golgi complex at 39 degrees C. The N protein was found evenly scattered in the cytoplasm, suggesting lack of interaction between the G1/G2 and N proteins. With ts6 and 11, G1 and G2 appeared to accumulate and aggregate in the endoplasmic reticulum (ER) at 39 degrees C. The location of the N protein coincided with that of the aggregated glycoproteins, suggesting that the N protein interacted with G1/G2 already in the ER. Thus, these mutants may prove valuable tools in studying the mechanism of Uukuniemi virus maturation.


The ductus arteriosus: a refined approach!
Although ongoing patency of the ductus arteriosus is common in small extremely preterm infants, consensus is lacking regarding its clinical significance and treatment strategies. Literature regarding likelihood of spontaneous closure, impact on neonatal morbidity and long-term outcomes, and adverse effects of intervention has led to uncertainty as to the best course of action. Enhancing the determination of hemodynamic significance and refining patient selection for therapeutic intervention will streamline the decision-making process. Targeted neonatal echocardiography performed by the clinician has gained popularity worldwide, and preliminary data show that it has the potential to optimize patient outcomes. We review the arguments for and against medical and surgical therapy, explore how targeted neonatal echocardiography used in conjunction with biomarkers may refine the treatment approach, and consider future directions in the field.


Effects of Housing First on Employment and Income of Homeless Individuals: Results of a Randomized Trial.
Housing First is emerging as an evidence-based practice for housing and supporting people who are homeless and have a mental illness. The objective of this study was to determine whether Housing First increases the odds of obtaining competitive employment in this population and affects income, including income from informal and illegal sources. A total of 2,148 people with a mental illness were recruited from five Canadian cities while they were homeless, classified as having moderate or high needs, and randomly assigned to Housing First or usual care. Housing First participants with high needs received assertive community treatment (ACT), and those with moderate needs received intensive case management (ICM). Every three months, participants were interviewed about employment and earnings in the previous months (median follow-up=745 days). Regression models were estimated via generalized estimating equations. ICM recipients had lower odds of obtaining employment compared with the control group with moderate needs. The odds of obtaining employment among ICM recipients increased but their employment rate never exceeded that of the control group. For ACT recipients, the odds of obtaining employment were not significantly different from those of the control group. Among Housing First participants, persons employed at baseline, men, and younger participants had greater odds of employment compared with control participants. Housing First did not appear to significantly increase income. This was the first large-scale randomized controlled study of Housing First's effects on employment. Further research is needed to determine how Housing First may be enhanced to increase odds of obtaining employment.


CT perfusion of renal cell carcinoma: impact of volume coverage on quantitative analysis.
To assess the feasibility, image quality, and radiation dose of computed tomography (CT) renal perfusion imaging in the adaptive 4-dimensional (4D)-spiral mode in patients with renal cell carcinoma (RCC), and to compare quantitative measurements between 2-dimensional regions-of-interest (2D-ROI) and 3-dimensional volumes-of-interest (3D-VOI). Twenty-one patients (13 male; age, 67.4 ± 9.5 years) with 24 histologically proven RCCs underwent CT perfusion imaging (100 kV, 100 mAs/rotation, scan range 10 cm, examination time 40.17 seconds) in a 4D-spiral mode with dual-source 128-slice CT. The ability to suspend respiration during CT perfusion imaging was visually monitored. Two independent readers assessed motion artifacts of CT perfusion imaging data sets on a 4-point scale before and after automated motion correction. Qualitative (enhancement pattern) and quantitative perfusion analysis (blood flow [BF], blood volume [BV], flow extraction product [KTrans]) were performed in the tumor and in healthy ipsi- and contralateral renal cortex applying the maximum-slope and a modified Patlak approach for quantitative analysis in 2D-ROI and 3D-VOI, the latter including the entire RCCs. Of the 21 patients, 8 (38%) could suspend respiration throughout the perfusion scan. Of 21 RCCs, 18 (86%) were completely included in the scan range. Motion artifacts were significantly reduced after automated motion correction (P < 0.001). All 24 RCCs could be included in the qualitative perfusion analysis, and 22 of 24 (92%) were eligible for quantitative perfusion analysis. Enhancement was homogenous in 4 (17%), peripheral in 4 (17%), and heterogeneous in 16 (66%) tumors (good interobserver agreement, κ=0.74). A high correlation was found between the 2 readers regarding quantitative perfusion parameters (r=0.93-0.94, P < 0.01). Quantitative measurements in 3D-VOIs revealed significantly lower BV, BF, and K in RCCs than in normal renal cortex (P < 0.001). In solid tumor periphery, BV was similar to the renal cortex (P=0.299), while BF and K were significantly lower (P < 0.01 and <0.001) in tumor tissue. Comparison of tumor measurements in 3D-VOIs with those obtained from 2D-ROIs revealed considerable differences in perfusion parameters beyond the 95% confidence limits in 46% to 68% of the tumors. KTrans was significantly higher in the contralateral than in healthy ipsilateral renal cortex (P < 0.01). Estimated effective radiation dose of the CT perfusion protocol was 16.3 mSv. CT perfusion imaging using an adaptive 4D-spiral mode is feasible and enables, after use of automated motion correction, the reliable analysis of renal perfusion in patients with RCCs. Considerable tumor heterogeneity was found, with differences in perfusion parameters between 2D-ROI and 3D-VOI analysis, reinforcing the use of volumetric techniques for perfusion imaging and analysis. Differences between ipsi- and contralateral healthy renal cortex KTrans suggest a compensatory increase in glomerular filtration rate in the healthy contralateral kidney.


What are the long-term consequences of youth exposure to firearm injury, and how do we prevent them? A scoping review.
The long-term consequences of exposure to firearm injury-including suicide, assault, and mass shootings-on children's mental and physical health is unknown. Using PRISMA-ScR guidelines, we conducted a scoping review of four databases (PubMed, Scopus, PsychINFO, and CJ abstract) between January 1, 1985 and April 2, 2018 for articles describing long-term outcomes of child or adolescent firearm injury exposure (n = 3582). Among included studies (n = 31), most used retrospective cohorts or cross-sectional studies to describe the correlation between firearm injury and post-traumatic stress. A disproportionate number of studies examined the effect of mass shootings, although few of these studies were conducted in the United States and none described the impact of social media. Despite methodologic limitations, youth firearm injury exposure is clearly linked to high rates of post-traumatic stress symptoms and high rates of future injury. Evidence is lacking on best practices for prevention of mental health and behavioral sequelae among youth exposed to firearm injury. Future research should use rigorous methods to identify prevalence, correlates, and intervention strategies for these at-risk youth.


Recent Advances in Understanding the Molecular Pathophysiology of Angiotensin II Receptors: Lessons From Cell-Selective Receptor Deletion in Mice.
The renin-angiotensin system (RAS) is an essential hormonal system involved in water and sodium reabsorption, renal blood flow regulation, and arterial constriction. Systemic stimulation of the RAS with infusion of the main peptide angiotensin II (Ang II) in animals as well as pathological elevation of renin (ie, renovascular hypertension) to increase circulatory Ang II in humans ultimately lead to hypertension and end organ damage. In addition to hypertension, accumulating evidence supports that the Ang II type 1 receptor exerts a critical role in cardiovascular and kidney diseases independent of blood pressure elevation. In the past 2 decades, the identification of an increased number of peptides and receptors has facilitated the concept that the RAS has detrimental and beneficial effects on the cardiovascular system depending on which RAS components are activated. For example, angiotensin 1-7 and Ang II type 2 receptors act as a counter-regulatory system against the classical RAS by mediating vasodilation. Although the RAS as an endocrine system for regulation of blood pressure is well established, there remain many unanswered questions and controversial findings regarding blood pressure regulation and pathophysiological regulation of cardiovascular diseases at the tissue level. This review article includes the latest knowledge gleaned from cell type-selective gene deleted mice regarding cell type-specific roles of Ang II receptors and their significance in health and diseases are discussed. In particular, we focus on the roles of these receptors expressed in vascular, cardiac, and kidney epithelial cells.


Oceans on titan?
If global oceans of methane exist on Titan, the atmosphere above them must be within 2 percent of saturation. The two Voyager radio occultation soundings, made at low latitudes, probably occurred over land, since they imply a relative humidity less, similar 70 percent near the surface. Oceans might exist at other low-latitude locations if the zonal wind velocities in the lowest 3 kilometers are </= 4 centimeters per second.


The Instructive Role of the Vasculature in Stem Cell Niches.
An important hallmark of many adult stem cell niches is their proximity to the vasculature in vivo, a feature common to neural stem cells (NSCs), mesenchymal stem cells (MSCs) from bone marrow, adipose, and other tissues, hematopoietic stem cells (HSCs), and many tumor stem cells. This review summarizes key studies supporting the vasculature's instructive role in adult stem cell niches, and the putative underlying molecular mechanisms by which blood vessels in these niches exert control over progenitor cell fates. The importance of the perivascular niche for pathology, notably tumor metastasis and dormancy, is also highlighted. Finally, the implications of the perivascular regulation of stem and progenitor cells on biomaterial design and the impact on future research directions are discussed.


Willingness to pay as a measure of the benefits of mental health care.
BACKGROUND: Decision-makers would benefit from being able to plan and evaluate mental health care interventions or programmes on the basis of costs and consequences that are measured in the same unit of measurement (money being the most convenient). Monetized quantification of the consequences of alternative interventions could be subsequently incorporated into cost-benefit allocation decisions. AIM: This paper provides an overview of the policy and research context within which willingness-to-pay survey techniques are located, together with a review of the main approaches used to date. We also highlight key issues in the application of these techniques and indicate areas of mental health research and policy that could benefit from their introduction. METHOD: Willingness-to-pay survey techniques are reviewed, and issues concerning their validity and application in the context of cost-benefit analyses of mental health policies are discussed. DISCUSSION: Different survey methods are available for generating willingness-to-pay data, the most common being the contingent valuation approach. An assessment of the validity of data generated by these alternative techniques is vital in order to ensure that they are consistent with the notion of economic preferences and values. IMPLICATIONS: The generation of valid and meaningful data on the monetized value of mental health outcomes would provide decision-makers with an improved evidence-based framework for resource allocation.


Cation-induced aggregation of membrane vesicles isolated from vascular smooth muscle.
Cations stimulated aortic muscle membrane aggregation with increasing potency according to their effective charge, e.g., K+ less than Mg2+ less than La3+, and the stimulation is reciprocally related to the apparent affinity for these cations. Divalent metal ion-induced membrane aggregation showed a dependence on the ionic radius, being optimal for Cd2+. Polyvalent cation-induced membrane aggregation was reversibly suppressed by high ionic strength as well as by metal ion chelators, irreversibly inhibited by the cross-linking agent glutaraldehyde, and enhanced by increasing concentrations of ethanol and increased temperature of the medium. When the pH is lowered below 6.0, membrane aggregation progressively increased with a concomitant decrease in cation-induced aggregation. The patterns of aggregation of microsomal membranes and further purified plasma membranes were almost identical whereas the aggregation of the heterogeneous mitochondrial membrane-enriched fraction was distinctly different in the initial rate of aggregation, its pH dependence, and metal ion concentration dependence. Our results indicate that cation-induced membrane aggregation can also be used to isolate a plasma membrane-enriched fraction from vascular smooth muscle.


Antimicrobial activity of sesquiterpene lactones isolated from traditional medicinal plant, Costus speciosus (Koen ex.Retz.) Sm.
Costus speciosus (Koen ex.Retz.) Sm (Costaceae) is an Indian ornamental plant which has long been used medicinally in traditional systems of medicine. The plant has been found to possess diverse pharmacological activities. Rhizomes are used to treat pneumonia, rheumatism, dropsy, urinary diseases, jaundice, skin diseases and leaves are used to treat mental disorders. Antibacterial and antifungal activities were tested using Disc diffusion method and Minimum Inhibitory Concentration (MIC). Column chromatography was used to isolate compounds from hexane extract. X-ray crystallography technique and GC-MS analysis were used to identify the compounds Antibacterial and antifungal activities were observed in hexane, chloroform, ethyl acetate and methanol extracts. Hexane extract of C.speciosus showed good activity against tested fungi also. Two sesquiterpenoid compounds were isolated (costunolide and eremanthin) from the hexane extract. Both the compounds did not inhibit the growth of tested bacteria. But, both the compounds inhibited the tested fungi. The compound costunolide showed significant antifungal activity. The MIC values of costunolide were; 62.5 μg/ml against Trichophyton mentagrophytes, 62. μg/ml against T. simii, 31.25 μg/ml against T. rubrum 296, 62.5 μg/ml against T. rubrum 57, 125 μg/ml against Epidermophyton floccosum, 250 μg/ml against Scopulariopsis sp, 250 μg/ml against Aspergillus niger, 125 μg/ml against Curvulari lunata, 250 μg/ml against Magnaporthe grisea. Hexane extract showed promising antibacterial and antifungal activity. The isolated compound costunolide showed good antifungal activity.


Microtubules, chromosome movement, and reorientation after chromosomes are detached from the spindle by micromanipulation.
The relationship between chromosome movement and microtubules was explored by combining micromanipulation of living grasshopper spermatocytes with electron microscopy. We detached chromosomes from the spindle and placed them far out in the cytoplasm. Soon, the chromosomes began to move back toward the spindle and the cells were fixed at a chosen moment. The microtubules seen in three-dimensional reconstructions were correlated with the chromosome movement just prior to fixation. Before movement began, detached chromosomes had no kinetochore microtubules or a single one at most. Renewed movement was always accompanied by the reappearance of kinetochore microtubules; a single kinetochore microtubule appeared to suffice. Chromosome movements and kinetochore microtubule arrangements were unusual after reattachment, but their relationship was not: poleward forces, parallel to the kinetochore microtubule axis (as in normal anaphase), would explain the movement, however odd. The initial arrangement of kinetochore microtubules would have led to aberrant chromosome distribution if it persisted, but instead, reorientation to the appropriate arrangement always followed. Observations on living cells permitted us to place in sequence the kinetochore microtubule arrangements seen in fixed cells, revealing the microtubule transformations during reorientation. From the sequence of events we conclude that chromosome movement can cause reorientation to begin and that in the changes which follow, an unstable attachment of kinetochore microtubules to the spindle plays a major role.


Co-implanting orthotopic tissue creates stroma microenvironment enhancing growth and angiogenesis of multiple tumors.
Tumor models are needed to study cancer. Noninvasive imaging of tumors under native conditions in vivo is critical but challenging. Intravital microscopy (IVM) of subcutaneous tumors provides dynamic, continuous, long-term imaging at high resolution. Although popular, subcutaneous tumor models are often criticized for being ectopic and lacking orthotopic tissue microenvironments critical for proper development. Similar IVM of orthotopic and especially spontaneous tumors is seldom possible. Here, we generate and characterize tumor models in mice for breast, lung, prostate and ovarian cancer by co-engrafting tumor spheroids with orthotopic tissue in dorsal skin window chambers for IVM. We use tumor cells and tissue, both genetically engineered to express distinct fluorescent proteins, in order to distinguish neoplastic cells from engrafted tissue. IVM of this new, two-colored model reveals classic tumor morphology with red tumor cell nests surrounded by green stromal elements. The co-implanted tissue forms the supportive stroma and vasculature of these tumors. Tumor growth and angiogenesis are more robust when tumor cells are co-implanted with orthotopic tissue versus other tissues, or in the skin alone. The orthotopic tissue promotes tumor cell mitosis over apoptosis. With time, tumor cells can adapt to new environments and ultimately even grow better in the non-orthotopic tissue over the original orthotopic tissue. These models offer a significant advance by recreating an orthotopic microenvironment in an ectopic location that is still easy to image by IVM. These "ectopic-orthotopic" models provide an exceptional way to study tumor and stroma cells in cancer, and directly show the critical importance of microenvironment in the development of multiple tumors.


LED therapy plus idebenone treatment targeting calcium and mitochondrial signaling pathways in dystrophic muscle cells.
Intracellular calcium dysregulation, oxidative stress, and mitochondrial dysfunction are some of the main pathway contributors towards disease progression in Duchenne muscular dystrophy (DMD). This study is aimed at investigating the effects of light emitting diode therapy (LEDT) and idebenone antioxidant treatment, applied alone or together in dystrophic primary muscle cells from mdx mice, the experimental model of DMD. Mdx primary muscle cells were submitted to LEDT and idebenone treatment and evaluated for cytotoxic effects and calcium and mitochondrial signaling pathways. LEDT and idebenone treatment showed no cytotoxic effects on the dystrophic muscle cells. Regarding the calcium pathways, after LEDT and idebenone treatment, a significant reduction in intracellular calcium content, calpain-1, calsequestrin, and sarcolipin levels, was observed. In addition, a significant reduction in oxidative stress level markers, such as H2O2, and 4-HNE levels, was observed. Regarding mitochondrial signaling pathways, a significant increase in oxidative capacity (by OCR and OXPHOS levels) was observed. In addition, the PGC-1α, SIRT-1, and PPARδ levels were significantly higher in the LEDT plus idebenone treated-dystrophic muscle cells. Together, the findings suggest that LEDT and idebenone treatment, alone or in conjunction, can modulate the calcium and mitochondrial signaling pathways, such as SLN, SERCA 1, and PGC-1α, contributing towards the improvement of the dystrophic phenotype in mdx muscle cells. In addition, data from the LEDT plus idebenone treatment showed slightly better results than those of each separate treatment in terms of SLN, OXPHOS, and SIRT-1.


Categorical scaling of duration bisection in pigeons (Columba livia), mice (Mus musculus), and humans (Homo sapiens).
A fundamental assumption underlying research in translational neuroscience is that animal models represent many of the same neurocognitive mechanisms and decision processes used by humans. Clear demonstrations of such correspondences will be crucial to the discovery of the neurobiological underpinnings of higher-level cognition. One domain likely to support fruitful comparisons is interval timing, because humans and other animals appear to share basic similarities in their ability to discriminate the durations of events in the seconds-to-minutes range. Here, we report that in a duration-bisection procedure using a series of anchor durations ranging from 2 through 5 s, pigeon, mouse, and human subjects classified a given signal duration as subjectively shorter than an adjacent, physically shorter signal duration when the two durations lay on opposite sides of a putative category boundary. These bisection reversals provide strong evidence for continuity of temporal cognition across a wide range of vertebrate species.


TRPV1 ablation aggravates inflammatory responses and organ damage during endotoxic shock.
To test the hypothesis that ablation of transient receptor potential vanilloid type 1 (TRPV1) channels leads to exacerbated inflammatory responses and organ damage during endotoxic shock, lipopolysaccharide (LPS; 5 million endotoxin units/kg of body weight) was injected intraperitoneally (i.p.) into wild-type (WT) and TRPV1-null mutant (TRPV1(-/-)) mice. Mean arterial pressure and heart rate, determined by radiotelemetry, were severely depressed after LPS injection into WT and TRPV1(-/-) mice, with no distinction between the two strains. At 24 h after LPS injection, renal glomerular hypercellularity and hepatocellular injury were observed in both strains, accompanying further elevated serum levels of creatinine and alanine aminotransferase in TRPV1(-/-) mice compared to those in WT mice. At 6 or 24 h after LPS injection, neutrophil recruitment into kidneys and livers, serum cytokine (tumor necrosis factor alpha [TNF-α], interleukin 1β [IL-1β], IL-6) and renal chemokine (KC, macrophage inflammatory protein 2 [MIP-2]) levels, and renal VCAM-1 and ICAM-1 expression were greater in TRPV1(-/-) mice than WT mice. In addition, increased plasma calcitonin gene-related peptide (CGRP) levels observed in WT mice 6 h after LPS injection were absent in TRPV1(-/-) mice. Thus, TRPV1 ablation aggravates inflammatory responses, including neutrophil infiltration, proinflammatory cytokine production, and adhesion molecule expression, leading to intensified organ damage during endotoxic shock in the absence of worsened circulatory failure. The data indicate that TRPV1 activation may attenuate endotoxin-induced organ damage, possibly via its anti-inflammatory action rather than alteration of hemodynamics.


Association of Fatigue and Outcomes in Advanced Cancer: An Analysis of Four SWOG Treatment Trials.
Patient-reported outcomes may be associated with cancer outcomes. We evaluated clinically significant fatigue (CSF), overall survival, adverse events (AEs), and quality of life (QOL) during cancer treatment. We compared outcomes in four phase II or III chemotherapy trials, two advanced non-small-cell lung cancer and two advanced hormone-refractory prostate cancer, with or without baseline CSF. CSF was defined as a rating of two or greater on the Functional Assessment of Cancer Therapy fatigue question or a European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-Core 30 fatigue symptom score of 50% or greater. Survival was compared according to CSF using Kaplan-Meier estimates and Cox regression models. Differences in AE rates by CSF were assessed via chi-squared tests, and QOL changes from baseline to 3 months via linear regression. Of 1,994 participants, 1,907 (median age 69 years, range: 32-91) had complete baseline QOL survey data, with 52% reporting CSF at baseline. For the two hormone-refractory prostate cancer studies, baseline CSF was associated with higher mortality rates, with adjusted hazard ratios of (95% CI, P value) 1.32 (1.13 to 1.55, P < .001) and 1.31 (1.02 to 1.67, P = .03) and with increased incidence of grade 3-5 constitutional (16.5% v 9.4%, P = .002; 13.9% v 6.3%, P = .002) and neurologic (11.7% v 6.1%, P = .006; 9.0% v 3.9%, P = .01) AEs, respectively. Baseline CSF was associated with a higher mortality rate in one non-small-cell lung cancer study: hazard ratio 1.44 and 1.04 to 2.00, P = .03. Oncology trial participants with baseline CSF had poorer survival and experienced more AEs than participants without CSF. This indicates fatigue as an important baseline prognostic factor in oncology treatment trials.


Prevalence and typing of human papilloma virus (HPV) among female sex workers and outpatient women in southern Thailand.
Thai sex workers (SW) have high rates of sexually transmitted infections (STIs); however, detection and treatment is often complicated by accessibility to sensitive and accurate diagnostic tests. Self-sampling of women combined with molecular amplification techniques could help in accurate diagnosis, treatment and follow-up of such women. Detection of persistent high-risk human papilloma virus (HPV) in such populations could also be beneficial in identifying women who may need more frequent follow-up for cervical cytology screening. The current study aimed to examine the prevalence of HPV in this population and compare this with the lower-risk outpatient women (OPW) in Thailand. Four hundred and thirty OPW and 524 Thai SW were sampled by a self-administered tampon collection. Cells were extracted from tampons and, subsequently, women underwent routine vaginal examination for detection of other STIs. Detection of HPV was performed by polymerase chain reaction (PCR) using the L1 consensus primers, followed by L1 consensus probe using an in-house PCR-enzyme-linked immunosorbent assay (ELISA). All positive samples were typed using PCR-ELISA and type-specific oligonucleotide probes. Overall, HPV was detected in 20/430 (4.6%) and 120/524 (22.9%) in OPW and SW respectively. Over 98% of samples produced a beta-globin signal, indicating adequately collected samples. Human papilloma virus typing probes detected HPV16 or 18 in 14% and 26% of the positive samples from OPW and SW respectively. HPV31, 33, 35 or 39 were detected in 19% and 12% and HPV45, 51 or 52 in 9.5% and 4% of positive OPW and SW patients respectively. Low risk HPV6 or 11 were detected in 1% and 4.9% of OPW and SW respectively. There was a significantly higher HPV prevalence in Thai SW than OPW, with the majority (21%) of positive samples containing the oncogenic HPV16 or 18 DNA. The results indicate that PCR could serve as a rapid and easy method for identification of women who require more frequent screening for cervical cancer.


Human gut microbiota does not ferment erythritol.
Erythritol, a naturally occurring polyol, is gaining attention as a bulk sweetener for human nutrition. Industrially, it is produced from glucose by fermentation. From various studies it is known to be non-cariogenic. Moreover, it is rapidly absorbed in the small intestine and quantitatively excreted in the urine. Only about 10 % enters the colon. Earlier in vitro experiments showed that erythritol remained unfermented for a fermentation period of 12 h. In order to investigate whether fresh human intestinal microbiota is able to adapt its enzyme activities to erythritol, a 24 h lasting fermentation was carried out under well-standardised in vitro conditions. For comparison maltitol, lactulose and blank (faecal inoculum only) were incubated as well. Fermentation patterns were established by following total gas production, hydrogen accumulation, changes in pH value, SCFA production and substrate degradation. Taking all fermentation parameters into account, erythritol turned out to be completely resistant to bacterial attack within 24 h, thus excluding an adaptation within that period. Since under in vivo conditions more easily fermentable substrates enter the colon continuously, it seems very unlikely that erythritol will be fermented in vivo.


Influence of copper on early development: prenatal and postnatal considerations.
Copper (Cu) is an essential nutrient whose requirement is increased during pregnancy and lactation. These represent times of critical growth and development, and the fetus and neonate are particularly vulnerable to deficiencies of this nutrient. Genetic mutations that predispose the offspring to inadequate stores of Cu can be life threatening as is observed in children with Menkes disease. During the last decade, severe Cu deficiency, once thought to be a rare condition, has been reported in the literature at an increasing frequency. Secondary Cu deficiencies can be induced by a variety of ways such as excessive zinc or iron intake, certain drugs, and bariatric surgery. Premature and low birth weight infants can be born with low Cu stores. A number of mechanisms can contribute to the teratogenicity of Cu including decreased activity of select cuproenzymes, increased oxidative stress, decreased nitric oxide availability, altered iron metabolism, abnormal extracellular matrix protein crosslinking, decreased angiogenesis and altered cell signaling among others. The brain, heart, and vessels as well as tissues such as lung, skin and hair, and systems including the skeletal, immune, and blood systems, are negatively affected by suboptimal Cu during development. Additionally, persistent structural, biochemical, and functional adverse effects in the offspring are noted even when Cu supplementation is initiated after birth, supporting the concept that adequate Cu nutriture during pregnancy and lactation is critical for normal development. Although Cu-containing IUDs are an effective method for increasing intrauterine Cu concentrations and for reducing the risk of pregnancy, high amounts of dietary Cu are not thought to represent a direct developmental risk.


Primary health care and donor dependency: a case study of nongovernment assistance in Burkina Faso.
Primary health care assistance has become prominent in the rural development programs of many nongovernment organizations throughout sub-Saharan Africa. By emphasizing education and the promotion of new participatory health systems, most such programs aim to enhance the conditions of women as principal community care givers. Yet village-level health assistance in Africa is not without shortcomings. This is exemplified in a case study of two nongovernment programs in Burkina Faso's Namentenga Province. Although both programs have contributed to maternal health and infant survival, they have also induced new ties of donor dependency. This appears to present a conundrum for the sponsoring agencies which espouse self-reliance as a development assistance goal. In fact, however, where the intervention of nongovernment organizations helps to improve rural health, new dimensions of dependency may prove to be a positive first stage in the mobilization of women and the development of locally managed health systems. For this to be so, much is contingent on the capacity of these organizations to integrate local participation in their own planning and management processes, and to augment the professional status of indigenous health workers.


Determination of chlorpyrifos-methyl, lambda-cyhalothrin and tebuconazole residues in Sultana seedless grapes sprayed with pesticides under farmer's conditions.
The aim of this study was to investigate pesticide residues on Sultana Seedless Grapes harvested at different times and consequently evaluate the risk. Analyses were performed with the Quick-Easy-Cheap-Efficient-Rugged-Safe (QuEChERS)-liquid chromatography/tandem mass spectrometry (LC-MS/MS) procedure. Pesticide-free grapes were spiked at 3 levels. Chlorpyrifos-methyl, lambda-cyhalothrin and tebuconazole detection limits were 20, 10 and 1 µg kg-1, respectively. These values were below maximum residue levels (MRL) of 1000, 80 and 500 µg kg-1, respectively. The overall recovery of the method was 108.60%. Present values were within acceptable recovery (60-140%) and repeatability (≤20%) ranges set by the Directorate-General for Health and Food Safety (SANTE). The vineyards were sprayed 4 times. Grapes were harvested in the 1st, 3rd, 5th, 7th and 14th days from the last spray. The residues in the samples taken from 5 vine stock groups were determined. MRL-exceeding chlorpyrifos-methyl residue of 1140.09 µg kg-1 was detected in the 1st day. The MRL-exceeding lambda-cyhalothrin (381.15 and 307.39 µg kg-1) and tebuconazole residues (650.58 and 570.85 µg kg-1) were detected in the 1st and 3rd days, respectively. The residues of 7th and 14th day samples were significantly different from the others. Excessive pesticide concentrations did not pose any health risks on consumers.


In-office, in-home, and telehealth cognitive processing therapy for posttraumatic stress disorder in veterans: a randomized clinical trial.
Trauma-focused psychotherapies for combat-related posttraumatic stress disorder (PTSD) in military veterans are efficacious, but there are many barriers to receiving treatment. The objective of this study was to determine if cognitive processing therapy (CPT) for PTSD among active duty military personnel and veterans would result in increased acceptability, fewer dropouts, and better outcomes when delivered In-Home or by Telehealth as compared to In-Office treatment. The trial used an equipoise-stratified randomization design in which participants (N = 120) could decline none or any 1 arm of the study and were then randomized equally to 1 of the remaining arms. Therapists delivered CPT in 12 sessions lasting 60-min each. Self-reported PTSD symptoms on the PTSD Checklist for DSM-5 (PCL-5) served as the primary outcome. Over half of the participants (57%) declined 1 treatment arm. Telehealth was the most acceptable and least often refused delivery format (17%), followed by In-Office (29%), and In-Home (54%); these differences were significant (p = 0.0008). Significant reductions in PTSD symptoms occurred with all treatment formats (p < .0001). Improvement on the PCL-5 was about twice as large in the In-Home (d = 2.1) and Telehealth (d = 2.0) formats than In-Office (d = 1.3); those differences were statistically large and significant (d = 0.8, 0.7 and p = 0.009, 0.014, respectively). There were no significant differences between In-Home and Telehealth outcomes (p = 0.77, d = -.08). Dropout from treatment was numerically lowest when therapy was delivered In-Home (25%) compared to Telehealth (34%) and In-Office (43%), but these differences were not statistically significant. CPT delivered by telehealth is an efficient and effective treatment modality for PTSD, especially considering in-person restrictions resulting from COVID-19. ClinicalTrials.gov ID NCT02290847 (Registered 13/08/2014; First Posted Date 14/11/2014).


Long-chain n-3 PUFA in vegetarian women: a metabolic perspective.
Vegetarian diets have been associated with health benefits, but paradoxically are low in EPA and DHA which are important for development, particularly of the central nervous system, and for health. Humans have limited capacity for synthesis of EPA and DHA from α-linolenic acid, although this is greater in women than men. Oily fish and, to a lesser extent, dairy foods and meat are the primary sources of EPA and DHA in the diet. Exclusion of these foods from the diet by vegetarians is associated consistently with lower EPA and DHA status in vegetarian women compared with omnivores. The purpose of the present review was to assess the impact of low EPA and DHA status in vegetarian pregnancies on the development and health of children. EPA and DHA status was lower in breast milk and in infants of vegetarian mothers than those born to omnivore mothers, which suggests that in the absence of pre-formed dietary EPA and DHA, synthesis from α-linolenic acid is an important process in determining maternal EPA and DHA status in pregnancy. However, there have been no studies that have investigated the effect of low maternal DHA status in vegetarians on cognitive function in children. It is important to address this gap in knowledge in order to be confident that vegetarian and vegan diets during pregnancy are safe in the context of child development.


Electron microscopy studies of alpha 2-macroglobulin subunit association after limited reduction with dithiothreitol.
The subunits of human alpha 2-macroglobulin (alpha 2M) were dissociated by treatment with reductant (0.5 mM dithiothreitol) under mild conditions. Intact tetramers, half-molecules (subunit dimers), and monomers were identified by chromatography on Superose-6. These products were not in rapidly reversible equilibrium since purified half-molecules were completely stable for up to 18 h. Monomers slowly associated to form some half-molecules in the same time period. Negatively stained preparations of monomers and half-molecules demonstrated significant heterogeneity by electron microscopy. This heterogeneity probably reflected structural differences as well as variation in projection and staining. After reaction with methylamine or proteinase, half-molecules and purified monomers reassociated. The principal product was an intact tetramer displaying an "H-like" image which was visually indistinguishable from the structure of intact tetrameric alpha 2M after conformational change. Incompletely reassociated alpha 2M species were also identified after performing chromatography to increase the fraction of these products. Images resembling one-half of the intact tetrameric alpha 2M structure (epsilon-image) or three-quarters of the intact tetramer (chair-image) were observed. These studies demonstrate that reassociation of reductant-dissociated alpha 2M subunits occurs by the formation of appropriate interactions, so that the resulting tetramers are equivalent to nontreated alpha 2M-trypsin. Unlike native alpha 2M, the structure of conformationally transformed alpha 2M is relatively insensitive to the loss of interchain disulfide bonds. We propose that reassembly of alpha 2M subunits occurs by the binding of two half-molecules or by the addition of individual subunits to half-molecules or trimers.


Recombinant human bone morphogenetic protein-2 in open tibial fractures. A subgroup analysis of data combined from two prospective randomized studies.
The use of recombinant human bone morphogenetic protein-2 (rhBMP-2) to improve the healing of open tibial shaft fractures has been the focus of two prospective clinical studies. The objective of the current study was to perform a subgroup analysis of the combined data from these studies. Two prospective, randomized clinical studies were conducted. A total of 510 patients with open tibial fractures were randomized to receive the control treatment (intramedullary nail fixation and routine soft-tissue management) or the control treatment and an absorbable collagen sponge impregnated with one of two concentrations of rhBMP-2. The rhBMP-2 implant was placed over the fracture at the time of definitive wound closure. For the purpose of this analysis, only the control treatment and the Food and Drug Administration-approved concentration of rhBMP-2 (1.50 mg/mL) were compared. Patients who anticipated receiving planned bone-grafting as part of a staged treatment were excluded from enrollment. Fifty-nine trauma centers in twelve countries participated, and patients were followed for twelve months postoperatively. Two subgroups were analyzed: (1) the 131 patients with a Gustilo-Anderson type-IIIA or IIIB open tibial fracture and (2) the 113 patients treated with reamed intramedullary nailing. The first subgroup demonstrated significant improvements in the rhBMP-2 group, with fewer bone-grafting procedures (p = 0.0005), fewer patients requiring invasive secondary interventions (p = 0.0065), and a lower rate of infection (p = 0.0234), compared with the control group. The second subgroup analysis of fractures treated with reamed intramedullary nailing demonstrated no significant difference between the control and the rhBMP-2 groups. The addition of rhBMP-2 to the treatment of type-III open tibial fractures can significantly reduce the frequency of bone-grafting procedures and other secondary interventions. This analysis establishes the clinical efficacy of rhBMP-2 combined with an absorbable collagen sponge implant for the treatment of these severe fractures.


Inositol 1,3,4,5-tetrakisphosphate stimulates calcium release from bovine adrenal microsomes by a mechanism independent of the inositol 1,4,5-trisphosphate receptor.
In bovine adrenal microsomes, Ins(1,4,5)P3 binds to a specific high-affinity receptor site (Kd = 11 nM) with low affinity for two other InsP3 isomers, Ins(1,3,4)P3 and Ins(2,4,5)P3. In the same subcellular fractions Ins(1,4,5)P3 was also the most potent stimulus of Ca2+ release of all the inositol phosphates tested. Of the many inositol phosphates recently identified in angiotensin-II-stimulated adrenal glomerulosa and other cells, Ins(1,3,4,5)P4 has been implicated as an additional second messenger that may act in conjunction with Ins(1,4,5)P3 to elicit Ca2+ mobilization. In the present study, an independent action of Ins(1,3,4,5)P4 was observed in bovine adrenal microsomes. Heparin, a sulphated polysaccharide which binds to Ins(1,4,5)P3 receptors in several tissues, inhibited both the binding of radiolabelled Ins(1,4,5)P3 and its Ca2(+)-releasing activity in adrenal microsomes. In contrast, heparin did not inhibit the mobilization of Ca2+ by Ins(1,3,4,5)P4, even at doses that abolished the Ins(1,4,5)P3 response. Such differential inhibition of the Ins(1,4,5)P3- and Ins(1,3,4,5)P4-induced Ca2+ responses by heparin indicates that Ins(1,3,4,5)P4 stimulates the release of Ca2+ from a discrete intracellular store, and exerts this action via a specific receptor site that is distinct from the Ins(1,4,5)P3 receptor.


Ultrastructure of multiple sclerosis.
The electron microscopic features of 11 stereotaxic brain biopsies that demonstrated inflammatory primary demyelination consistent both morphologically and clinically with multiple sclerosis are addressed. Degeneration of inner oligodendroglial loops and uniform widening of inner myelin lamellae antedated complete destruction of myelin sheaths. Perivascular lymphocytes, macrophages, and plasma cells were in intimate contact with myelin sheaths. Astrocytes proliferated even away from demyelinated areas. In areas of chronic, established demyelination, oligodendrocyte numbers were greatly decreased, and fields of completely demyelinated axons were seen among astrocytic processes. Axonal injury, evidenced by the formation of axonal swellings, was apparent in maximally affected areas. At the edge of acute lesions with demyelinated axons, oligodendrocytes were preserved morphologically. Thinly myelinated axons indicative of central nervous system-type remyelination by oligodendrocytes were observed primarily at the edges of plaques. An unusual inclusion observed in presumed macrophages was "polelike" bodies 0.04- to 0.7-microns thick. Linearly arrayed, their presumably proteinaceous crystalline substance was moderately electron-dense. Many were membrane-bound and appeared to arise from the endoplasmic reticulum. We conclude that disturbance of the myelinating function of oligodendrocytes may be a critical event in the pathogenesis of multiple sclerosis.


Ex vivo confocal microscopy performs real-time assessment of renal biopsy in non-neoplastic diseases.
Ex vivo confocal microscopy is a technique for tissue examination, which generates images of fresh samples with an optical resolution comparable to those obtained by conventional pathology. The objective of this study was to evaluate the feasibility of using ex vivo confocal microscopy in fusion mode (reflectance and fluorescence) and the H&E-like digital staining that is obtained for the analysis of non-neoplastic kidney biopsies. Twenty-four renal samples acquired from autopsies were scanned in a 4th generation ex vivo confocal microscopy device. The imaging process was completed in an average of three minutes. Confocal images correlated very well to the corresponding conventional histological sections, both in normal tissue and in chronic lesions (glomerulosclerosis, fibrosis and tubular atrophy). The ex vivo confocal microscopy protocol did not add artifacts to the sample for the ulterior study with light microscopy, nor to the histochemical or immunohistochemical studies. The ease and speed of grayscale and fluorescence image acquisition, together with the quality of the H&E-like digitally stained images obtained with this approach, suggest that this technique shows promise for use in clinical nephrology and renal transplantation.


Development of the Implantable Cardioverter Defibrillator Body Image Concerns Questionnaire.
The implantable cardioverter defibrillator (ICD) is the first-line treatment for the prevention of sudden cardiac death. Although most ICD recipients adapt well to living with the device, some struggle with ICD-related body image concerns (BICs). Because of lack of standardized questionnaires to assess BICs, we do not know the prevalence. The aim of this study was to develop a questionnaire that assesses self-reported ICD-related BICs. Using a sequential, qualitative multimethod approach, we developed the ICD-BIC questionnaire (ICD-BICQ) based on (1) themes from a scoping review on BICs in ICD recipients, (2) cognitive interviews with ICD recipients to evaluate the validity of items and to ascertain whether items were missing, and (3) input from healthcare professionals. A conceptual framework was created containing 4 general dimensions: (1) assessment, (2) behavior, (3) body perception, and (4) emotions. Implantable cardioverter defibrillator recipients (n = 8, 2 women and 6 men; age, 40-78 years), 2 specialists in questionnaire development, and healthcare professionals specialized in ICD treatment evaluated the questionnaire. After no new items or new dimensions emerged from the cognitive interviews, the final version 7 of the ICD-BICQ consisted of 39 items tapping into (1) assessment (n = 2), (2) behavior (n = 13), (3) body perception (n = 16), and emotions (n = 8). Items were assessed on a 5-point Likert-type scale. Using patient and healthcare professionals' involvement, we developed the 39-item ICD-BICQ to measure the prevalence of BICs in ICD recipients. The factor structure, construct validity, psychometric properties, and a clinically relevant cutoff for the ICD-BICQ will be evaluated in a quantitative study of ICD recipients.


ANTIBODY RESPONSE TO IMMUNIZATION BY DIFFERENT ROUTES.
Rabbits were injected with human serum albumin once intravenously or intradermally with this antigen plus complete Freund adjuvant. The hemagglutinating titers of the antiserum and of the heavy and light antibody fractions were followed for several months. Circulation of heavy antibody in the intravenously injected group disappeared in 1 month, but in the intradermally injected group, it rose to a maximum titer, then declined, and then showed a second rise in titer. Light antibody showed increasing and then sustained titers over most of the time period. Heavy antibody failed to precipitate with antigen, even when it showed high hemagglutinating titers. Light antibody, in contrast, gave precipitation even when tested at much lower hemagglutination titers


Intragenic recombination and a chimeric outer membrane protein in the relapsing fever agent Borrelia hermsii.
The spirochete Borrelia hermsii, a relapsing fever agent, evades the host's immune response through multiphasic antigenic variation. Antigen switching results from sequential expression of genes for serotype-specific outer membrane proteins known as variable major proteins (Vmp's); of the 25 serotypes that have been identified for the HS1 strain, serotypes 7 and 21 have been studied in greatest detail. In the present study, an atypical variant was predominant in the relapse from a serotype 21 infection in mice; relapse cells were bound by monoclonal antibodies specific for Vmp21 as well as antibodies specific for Vmp7. In Western blots (immunoblots), the variant had a single Vmp that was reactive with monoclonal antibodies representing both serotypes. The gene encoding this Vmp, vmp7/21, was cloned and characterized by restriction mapping and sequence analysis to determine the likely recombination event. Whereas the 5' end of vmp7/21 was identical to that of vmp21, its 3' end and flanking sequences were identical to the 3' end of vmp7. Unlike other vmp genes examined thus far, the vmp7/21 gene existed only in an expressed form; a silent, storage form of the gene was not detected. We conclude that the vmp7/21 gene was created by an intragenic recombination between the formerly expressed vmp21 gene and a silent vmp7 gene. This finding suggests that the lack of cross-reactivity between variants, which is usually observed, results from immunoselection against variants possessing chimeric Vmp's rather than from a switching mechanism that excludes partial gene replacements.


Post-mortem clinical pharmacology.
Clinical pharmacology assumes that deductions can be made about the concentrations of drugs from a knowledge of the pharmacokinetic parameters in an individual; and that the effects are related to the measured concentration. Post-mortem changes render the assumptions of clinical pharmacology largely invalid, and make the interpretation of concentrations measured in post-mortem samples difficult or impossible. Qualitative tests can show the presence of substances that were not present in life, and can fail to detect substances that led to death. Quantitative analysis is subject to error in itself, and because post-mortem concentrations vary in largely unpredictable ways with the site and time of sampling, as a result of the phenomenon of post-mortem redistribution. Consequently, compilations of 'lethal concentrations' are misleading. There is a lack of adequate studies of the true relationship between fatal events and the concentrations that can be measured subsequently, but without such studies, clinical pharmacologists and others should be wary of interpreting post-mortem measurements.


[Investigation of growth prognosis in asthmatic children].
The objective of the present study was the ascertainment of correct data related to growth prognosis in asthmatic children. For this purpose, 210 young asthmatics including 153 males and 57 females ranging in age from 1.7 to 18.9 years were evaluated. Data were specified for the parameters: chronological age (CA); bone age (KA); height age (LA); height (KH); severity of illness; target adult height (GZG) and predicted adult height (PEG). Among this group, 5.9 % of the boys and 3.5 % of the girls, were found to be of small stature (KH < - 2 s). This corresponds to about 2.5 and 1.5 times the average in a normal distribution. Children with the highest severity of illness showed the strongest negative deviation for the KH-SDS values (boys - 0.59 plus minus 1.1, girls - 0.97 plus minus 0.8). This proved to be statistically significant for females. Development of CA, KA and LA continued unremarkably until the age of 4 years. From age 5, partially a significant growth retardation of KA and LA could be observed in both sexes. To age 16, KA values and, after that LA values, were more strongly affected. Comparison of the GZG and PEG average values within the degree of severity groups made it clear that the PEG values for both sexes, independent of the severity of illness, were without exception, smaller than the GZG. The differences of both parameters proved to be statistically significant for the group with the most severe symptoms of the boys (180.8 plus minus 6.5 cm to 175.8 plus minus 6.2 cm, p < 0.01) and for the entire groups of both sexes (boys 180.2 plus minus 5.3 cm to 178.6 plus minus 7.3 cm, p < 0.05, girls 167.7 plus minus 4.4 cm to 165.8 plus minus 6.8 cm, p < 0.05). However, a separate comparison of GZG and PEG average values, respectively, among the severity groups showed no significant differences. The results argue for a growth-inhibiting influence from bronchial asthma. The cause has to be severity of illness but direct effects of the atopy on skeletal development is also taken in consideration. Confirmation of these findings requires investigation of a larger group of asthmatics and clarification of the pathophysiological processes in the growing skeletons of atopic children.


The major determinant of attenuation in mice of the Candid1 vaccine for Argentine hemorrhagic fever is located in the G2 glycoprotein transmembrane domain.
Candid1, a live-attenuated Junin virus vaccine strain, was developed during the early 1980s to control Argentine hemorrhagic fever, a severe and frequently fatal human disease. Six amino acid substitutions were found to be unique to this vaccine strain, and their role in virulence attenuation in mice was analyzed using a series of recombinant viruses. Our results indicate that Candid1 is attenuated in mice through a single amino acid substitution in the transmembrane domain of the G2 glycoprotein. This work provides insight into the molecular mechanisms of attenuation of the only arenavirus vaccine currently available.


Guiding principles for peptide nanotechnology through directed discovery.
Life's diverse molecular functions are largely based on only a small number of highly conserved building blocks - the twenty canonical amino acids. These building blocks are chemically simple, but when they are organized in three-dimensional structures of tremendous complexity, new properties emerge. This review explores recent efforts in the directed discovery of functional nanoscale systems and materials based on these same amino acids, but that are not guided by copying or editing biological systems. The review summarises insights obtained using three complementary approaches of searching the sequence space to explore sequence-structure relationships for assembly, reactivity and complexation, namely: (i) strategic editing of short peptide sequences; (ii) computational approaches to predicting and comparing assembly behaviours; (iii) dynamic peptide libraries that explore the free energy landscape. These approaches give rise to guiding principles on controlling order/disorder, complexation and reactivity by peptide sequence design.


Inhibitory effect of tumor suppressor p33(ING1b) and its synergy with p53 gene in hepatocellular carcinoma.
To investigate the inhibitory effect of tumor suppressor p33(ING1b) and its synergy with p53 gene in hepatocellular carcinoma (HCC). Recombinant sense and antisense p33(ING1b) plasmids were transfected into hepatoma cell line HepG2 with lipofectamine. Apoptosis, G0/G1 arrest, cell growth rate and cloning efficiency in soft agar of HepG2 were analyzed after transfection. In three hepatoma cell lines with different endogenous p53 gene expressions, the synergistic effect of p33(ING1b) with p53 was analyzed by flow cytometry and luciferase assay was performed to detect the activation of p53 downstream gene p21(WAF1/CIP1). In addition, the expression and mutation rates of p33(ING1b) in HCC tissues were measured by immunohistochemistry and polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). Overexpression of p33(ING1b) inhibited cell growth of HepG2, induced more apoptosis and protected cells from growth in soft agar. Combined transfer of p33(ING1b) and p53 gene promoted hepatoma cell apoptosis, G0/G1 arrest and elevated expression of p21(WAF1/CIP1). Immunostaining results showed co-localized P33(ING1b) with P53 protein in HCC tissues and there was a significant relation between protein expression rates of these two genes (P<0.01). Among 28 HCC samples, p33(ING1b) presented a low gene mutation rate (7.1%). p33(ING1b) collaborates with p53 in cell growth inhibition, cell cycle arrest and apoptosis in HCC. Loss or inactivation of p33(ING1b) normal function may be an important mechanism for the development of HCC retaining wild-type p53.


Sound rhythms are encoded by postinhibitory rebound spiking in the superior paraolivary nucleus.
The superior paraolivary nucleus (SPON) is a prominent structure in the auditory brainstem. In contrast to the principal superior olivary nuclei with identified roles in processing binaural sound localization cues, the role of the SPON in hearing is not well understood. A combined in vitro and in vivo approach was used to investigate the cellular properties of SPON neurons in the mouse. Patch-clamp recordings in brain slices revealed that brief and well timed postinhibitory rebound spiking, generated by the interaction of two subthreshold-activated ion currents, is a hallmark of SPON neurons. The I(h) current determines the timing of the rebound, whereas the T-type Ca(2+) current boosts the rebound to spike threshold. This precisely timed rebound spiking provides a physiological explanation for the sensitivity of SPON neurons to sinusoidally amplitude-modulated (SAM) tones in vivo, where peaks in the sound envelope drive inhibitory inputs and SPON neurons fire action potentials during the waveform troughs. Consistent with this notion, SPON neurons display intrinsic tuning to frequency-modulated sinusoidal currents (1-15Hz) in vitro and discharge with strong synchrony to SAMs with modulation frequencies between 1 and 20 Hz in vivo. The results of this study suggest that the SPON is particularly well suited to encode rhythmic sound patterns. Such temporal periodicity information is likely important for detection of communication cues, such as the acoustic envelopes of animal vocalizations and speech signals.


A Complication Profile of Total Hip and Knee Arthroplasty in Liver Transplantation Patients: A Meta-Analysis.
There is an increasing demand for total joint arthroplasty in liver transplantation patients. However, significant heterogeneity in existing studies creates difficulty to draw conclusions on the risk profile of arthroplasty in this population. A systematic review of the literature dated from 1980 to 2020 describing the complication rates of liver transplantation patients receiving either total hip or knee arthroplasty was conducted. Multiple outcomes were extracted and a meta-analysis was performed. Four cohorts were created for analysis purposes: liver transplant patients undergoing THA and TKA (1), THA only (2), TKA only (3), and controls (4). A total of 13 studies were included in this meta-analysis, accounting for 3024 liver transplantation patients. The rate of infection (odds ratio [OR] = 2.14, OR = 1.61, OR = 2.52), myocardial infarction (OR = 1.65, OR = 1.75, OR = 1.57), respiratory failure (OR = 2.19, OR = 2.50, OR = 1.96), acute kidney injury (OR = 5.71, OR = 5.40, OR = 4.35), sepsis (OR = 3.72, OR = 3.30, OR = 4.02), and blood transfusions (OR = 2.09, OR = 3.65, OR = 1.74) were all significantly higher in the 3 cohorts compared to the controls. Revision/reoperation rates were significantly higher in cohorts 1 and 3 (OR = 1.52 and OR = 1.62, respectively). Patient-reported outcomes saw improvements in Harris Hip Score, objective Knee Society Score, and functional Knee Society Score postoperatively (average improvement = 32.4, 37.2, and 15.3, respectively). Liver transplantation patients functionally benefit from total hip and knee arthroplasty, but at the cost of increased risk of infection, revision/reoperation, and medically related complications compared to controls. Mortality may also be a short-term risk.


Hybrids formed by fusion of X- or gamma-irradiated and non-irradiated mouse cells.
Hybrid cells were obtained by fusion of irradiated and non-irradiated mouse cells of two different lines; they differed from the parent lines and from the hybrid cells of non-irradiated parents in their morphological, growth and karyological properties. The frequency of their occurrence was lower than in hybrids from non-irradiated cells, and unlike the irradiated cells of the parent line, these hybrid cells were capable of permanent proliferation in vitro. Chromosomes of the irradiated parent line were preferentially eliminated from the karyotype of the hybrids.


Arch development with trans-force lingual appliances.
Trans-Force lingual appliances are designed to correct arch form in patients with contracted dental arches. Interceptive treatment with this new series of pre-activated lingual appliances offers new possibilities for arch development, in combination with fixed appliances. Palatal and lingual appliances insert in horizontal lingual sheaths in molar bands. No activation is required after the appliance is fitted, and this principle is extended to a series of appliances for sagittal and transverse arch development. Both sagittal and transverse appliances have additional components to achieve 3-way expansion where this is indicated. The invisible lingual appliances may be used in correction of all classes of malocclusion at any stage of development, from mixed dentition through permanent dentition, and this approach has wide indications in adult treatment.


Inhibition of Heme Oxygenase Antioxidant Activity Exacerbates Hepatic Steatosis and Fibrosis In Vitro.
The progression of non-alcoholic fatty liver disease (NAFLD) and the development of hepatic fibrosis is caused by changes in redox balance, leading to an increase of reactive oxygen species (ROS) levels. NAFLD patients are at risk of progressing to non-alcoholic steatohepatitis (NASH), associated to cardiovascular diseases (CVD), coronary heart disease and stroke. Heme Oxygenase-1 (HO-1) is a potent endogenous antioxidant gene that plays a key role in decreasing oxidative stress. The present work was directed to determine whether use of an inhibitor of HO-1 activity affects lipid metabolism and fibrosis process in hepatic cells. Oil Red assay and mRNA analysis were used to evaluate the triglycerides content and the lipid metabolism pathway in HepG2 cells. ROS measurement, RT-PCR and Soluble collagen assay were used to assess the intracellular oxidant, the fibrosis pathway and the soluble collagen in LX2 cells. The activity of HO-1 was inhibited using Tin Mesoporphyrin IX (SnMP). Our study demonstrates that a non-functional HO system results in an increased lipid storage and collagen release in hepatocytes. Consequently, an increase of HO-1 levels may provide a therapeutic approach to address the metabolic alterations associated with NAFLD and its progression to NASH.


A Novel Cell Membrane-Associated RNA Extraction Method and Its Application in the Discovery of Breast Cancer Markers.
Cell membrane-associated RNA (mem-RNA) has been demonstrated to be cell-specific and disease-related and are considered as potential biomarkers for disease diagnostics, drug delivery, and cell screening. However, there is still a lack of methods specifically designed to extract mem-RNA from cells, limiting the discovery and applications of mem-RNA. In this study, we propose the first all-in-one solution for high-purity mem-RNA isolation based on two types of magnetic nanoparticles, named MREMB (Membrane-associated RNA Extraction based on Magnetic Beads), which achieved ten times enrichment of cell membrane components and over 90% recovery rate of RNA extraction. To demonstrate MREMB's potential in clinical research, we extracted and sequenced mem-RNA of typical breast cancer MCF-7, MDA-MB-231, and SKBR-3 cell lines and non-neoplastic breast epithelial cell MCF-10A. Compared to total RNA, sequencing results revealed that membrane/secreted protein-encoding mRNAs and long noncoding RNAs (lncRNAs) were enriched in the mem-RNA, some of which were significantly overexpressed in the three cancer cell lines, including extracellular matrix-related genes COL5A1 and lncRNA TALAM1. The results indicated that MREMB could enrich membrane/secreted protein-coding RNA and amplify the expression differences of related RNAs between cancer and non-neoplastic cells, promising for cancer biomarker discovery.


Candidate genes for insulin resistance.
Insulin resistance confers increased susceptibility to NIDDM, atherosclerotic cardiovascular disease, ovarian hyperandrogenism, and possibly hypertension. Insulin resistance is largely inherited, in rare cases as a monogenic disorder or more commonly as a complex trait. The search for insulin resistance genes relies mainly on two complementary approaches: 1) positional cloning using random DNA markers present throughout the genome; and 2) the analysis of specific candidate genes. This report briefly summarizes the candidate gene approach to insulin resistance. Progress related to the analysis of genes encoding molecules that participate in insulin action is reviewed. In addition, the spectrum of potential genetic defects that might contribute to insulin resistance, both at the level of the target cell and secondarily (e.g., obesity genes), is discussed.


Is the 2008 NMC Code ethical?
In 2008 the United Kingdom Nursing and Midwifery Council (NMC) published the latest version of its code of conduct (The code: standards of conduct, performance and ethics for nurses and midwives). The new version marked a significant change of style in the Code compared with previous versions. There has been considerable controversy and the accrual of an extensive body of literature over the years in the UK and Europe criticizing nursing codes of ethics and questioning their ethical standing and their usefulness. In this article we review the current NMC Code. We argue that the NMC has been misguided in labelling the Code as a code of ethics, and suggest that the new document falls short in many respects.


Targeting Cognition and Motivation in Coordinated Specialty Care for Early Psychosis: A Grant Report.
In this grant report, we describe our project to expand measurement-based psychiatric care across 6 early psychosis treatment teams in Minnesota, and to provide a neuroscience-informed cognitive training and motivation enhancement program for individuals with early psychosis. This project is part of the NIMH Early Psychosis Intervention Network (EPINET) initiative which seeks to link data from treatment centers nationally that offer evidence-based specialty care to persons experiencing early psychosis. Systematic analyses of pooled data collected in EPINET will help inform methods for early psychosis care, psychosis risk factors, and pre-emptive interventions. As part of the national EPINET, our hub (Early Psychosis Intervention-Minnesota, EPI-MINN), will: (1) provide measurement-based care in coordinated specialty care programs for early psychosis, (2) determine whether a structured feedback report provides benefit to stakeholders-service users, family members, and primary clinicians, and (3) explore whether deficits in cognition and motivated behavior-two domains that significantly impact functioning and overall quality of life in early psychosis-can be addressed as key treatment goals by implementing a 12-week mobile intervention. Using a regression discontinuity design, participants will be randomized to the cognitive training and motivational enhancement intervention or to treatment as usual. The intervention consists of neuroscience-informed, computerized auditory and social cognitive training exercises, as well as a mobile app where participants interact with each other and with a motivational coach. Participants will complete assessments at 4 time points: baseline and post-intervention (i.e., at 6 months), and again at 12 and 18 months to test the long-term effects of the intervention. All assessments and interventions in this project can be completed entirely remotely.


Significant association of insulin and proinsulin with clustering of cardiovascular risk factors.
To investigate the association between true insulin and proinsulin and clustering of cardiovascular risk factors. Based on the random stratified sampling principles, 1196 Chinese people (533 males and 663 females, aged 35-59 years with an average age of 46.69 years) were recruited. Biotin-avidin based double monoclonal antibody ELISA method was used to detect the true insulin and proinsulin, and a risk factor score was set to evaluate individuals according to the number of risk factors. The median (quartile range) of true insulin and proinsulin was 4.91 mIu/L (3.01-7.09 mIu/L) and 3.49 pmol/L (2.14-5.68 pmol/L) respectively, and the true insulin level of female subjects was significantly higher than that of male subjects (P = 0.000), but the level of proinsulin displayed no significant difference between males and females (P = 0.566). The results of covariate ANOVA after age and sex were controlled showed that subjects with any of the risk factors had a significantly higher true insulin level (P = 0.002 for hypercholesterolemia, P = 0.021 for high low-density lipoprotein cholesterol, P = 0.003 for low high-density lipoprotein cholesterol, and P = 0.000 for other risk factors) and proinsulin level (P = 0.001 for low high-density lipoprotein cholesterol, and P = 0.000 for other risk factors) than those with no risk factors. Furthermore, subjects with higher risk factor scores had a higher true insulin and proinsulin level than those with lower risk factor scores (P = 0.000). The multiple linear regression models showed that true insulin and proinsulin were significantly related to cardiovascular risk factor scores respectively (P = 0.000). True insulin and proinsulin are significantly associated with the clustering of cardiovascular risk factors.


Modern biotechnology-based therapeutic approaches against HIV infection.
The causative agent of acquired immune deficiency syndrome (AIDS) is human immunodeficiency virus (HIV). Since its discovery before 30 years, a number of drugs known as highly active antiretroviral therapy have been developed to suppress the life cycle of the virus at different stages. With the current therapeutic approaches, ending AIDS means providing treatment to 35 million individuals living with HIV for the rest of their lives or until a cure is developed. Additionally, therapy is associated with various other challenges such as potential of drug resistance, toxicity and presence of latent viral reservoir. Therefore, it is imperative to search for treatments and to identify new therapeutic approaches against HIV infection to avoid daily intake of drugs. The aim of the current review was to summarize different therapeutic strategies against HIV infection, including stem cell therapy, RNA interference, CRISPR/Cas9 pathways, antibodies, intrabodies and nanotechnology. Silencing RNA against chemokine receptor 5 and other HIV RNAs have been tested and found to elicit homology-based, post-transcriptional silencing. The CRISPR/Cas9 is a gene editing technology that produces a double-stranded nick in the virus DNA, which is repaired by the host machinery either by non-homology end joining mechanism or via homology recombination leading to insertion, deletion mutation which further leads to frame shift mutation and non-functional products. Intrabodies are intracellular-expressed antibodies that are directed towards the targets inside the cell unlike the naturally expressed antibodies which target outside the cell. Different nanotechnology-based therapeutic approaches are also in progress against HIV. HIV eradication is not feasible without deploying a cure or vaccine alongside the treatment.


Consensus in acute myeloid leukemia in Mexico.
Acute myeloid leukemia (AML) comprises a heterogeneous group of hematopoietic cell neoplasms of myeloid lineage that arise from the clonal expansion of their precursors in the bone marrow, interfering with cell differentiation, leading to a syndrome of bone marrow failure. AML is a consequence of genetic and epigenetic changes (point mutations, gene rearrangements, deletions, amplifications, and arrangements in epigenetic changes that influence gene expression) in hematopoietic precursor cells, which create a clone of abnormal cells that are capable of proliferating but cannot differentiate into mature hematopoietic cells or undergo programmed cell death. The diagnosis requires more than 20% myeloid blasts in the bone marrow and certain cytogenic abnormalities. Treatment will depend on age, comorbidities, and cytogenetic risk among the most frequent.


ACR Appropriateness Criteria® assessment of gravid cervix.
It is well recognized that preterm birth is the leading cause of perinatal mortality and morbidity. There is a significant association between cervix length and preterm birth risk. Most authorities consider a cervical length <3 cm as the lower limit of normal. A cervical length >3 cm has a high negative predictive value for delivery less than 34 weeks. A cervical length of <15 mm is moderately predictive (∼ 70%) of preterm birth within 48 hours. Cervical length is normally distributed and should remain relatively constant until the third trimester. Transabdominal US is the least reliable method of cervical length assessment. The most reliable method of documenting cervical length is transvaginal ultrasound (TVUS). Transperineal US is an alternative for imaging if TVUS is contraindicated, such as with premature rupture of membranes. However, the resolution is decreased compared to TVUS. Short cervix length is the single most important predictive finding for premature delivery. This observation should prompt consultation for high risk obstetrical care and consideration of other management options such as cerclage or activity restriction.The ACR Appropriateness Criteria are evidence-based guidelines for specific clinical conditions that are reviewed biennially by a multidisciplinary expert panel. The guideline development and review include an extensive analysis of current medical literature from peer reviewed journals and the application of a well-established consensus methodology (modified Delphi) to rate the appropriateness of imaging procedures by the panel. In those instances where evidence is lacking or not definitive, expert opinion may be used to recommend imaging.


Purification of combinatorial libraries.
In the early days of combinatorial chemistry, much attention focused on preparation of large libraries for lead discovery. Recently, though, the focus has shifted toward smaller, more focused libraries for lead optimization. These focused libraries generally consist of individual discrete compounds. Biological assay requirements often require compounds of high purity, thus development of automated high throughput purification methods has received new attention in the past several years. This paper covers automated high throughput purification methods that have been applied to libraries of discrete compounds. Literature published through February 1998 is included. Purification methods discussed include extraction methods, scavenger methods, solid phase extraction, and preparative HPLC.


Spontaneous regression of a vein of Galen aneurysmal malformation in a pediatric patient: illustrative case.
Vein of Galen aneurysmal malformations (VGAMs) are rare congenital intracranial vascular lesions that represent 30% of all pediatric vascular anomalies. These lesions are associated with severe manifestations, including congestive heart failure, hydrocephalus, and spontaneous hemorrhage. The mainstay of management is medical stabilization followed by endovascular embolization of the lesion. Although VGAM was first reported in 1937, there are few published cases demonstrating spontaneous regression of the lesion. The authors report the case of a 31-month-old female who presented with an incidentally found VGAM. After initial evaluation, including magnetic resonance imaging and angiography, the patient was lost to follow-up. Upon her return to the clinic at age 12 years, the previously identified VGAM was absent, indicative of involution of the lesion. The patient remained asymptomatic and met appropriate developmental milestones during this interval. This report adds a rare case of the spontaneous resolution of VGAM to the literature. This case may suggest the presence of VGAMs that are asymptomatic, undetected, and regress within the pediatric population. Future studies may benefit from identifying imaging and angiographic findings predictive of spontaneous regression. There may be a role for conservative management in particular cases of asymptomatic and medically stable children with VGAMs.


The P2Y2/Src/p38/COX-2 pathway is involved in the resistance to ursolic acid-induced apoptosis in colorectal and prostate cancer cells.
One of the hallmarks of cancer is resistance to apoptosis. Elucidating the mechanisms of how cancer cells evade or delay apoptosis should lead to novel therapeutic strategies. Previously, we showed that HT-29 colorectal cancer cells undergoing apoptosis overexpressed cyclooxygenase-2 (COX-2), in a p38 dependent pathway, to delay ursolic acid-induced apoptosis. Here, we focused on elucidating the upstream signaling pathways regulating this resistance mechanism. The role of ATP as an extracellular signaling molecule took a long time to be accepted. In recent years, ATP and its analogs, via the activation of specific purinergic receptors, have been implicated in many biological processes including cell proliferation, differentiation and apoptosis. In the present report, we have demonstrated a novel role involving purinergic receptors and particularly the P2Y(2) receptor in resistance to ursolic acid-induced apoptosis in both colorectal HT-29 and prostate DU145 cancer cells. We found that ursolic acid induced an increase in intracellular ATP and P2Y(2) transcript levels. Upon activation, P2Y(2) activated Src which in turn phosphorylated p38 leading to COX-2 overexpression which induced resistance to apoptosis in both HT-29 and DU145 cells. Furthermore, Ca(2+)-independent PLA(2) (iPLA(2)) and Ca(2+)-dependent secretory PLA(2) (sPLA(2)) were responsible for arachidonic acid release, the substrate of COX-2. Our findings document that apoptosis triggering was dependent on protein kinase C (PKC) activation in both cell lines after ursolic acid treatment.