The complex hybrid origins of the root knot nematodes revealed through comparative genomics.
Root knot nematodes (RKN) can infect most of the world's agricultural crop species and are among the most important of all plant pathogens. As yet however we have little understanding of their origins or the genomic basis of their extreme polyphagy. The most damaging pathogens reproduce by obligatory mitotic parthenogenesis and it has been suggested that these species originated from interspecific hybridizations between unknown parental taxa. We have sequenced the genome of the diploid meiotic parthenogen Meloidogyne floridensis, and use a comparative genomic approach to test the hypothesis that this species was involved in the hybrid origin of the tropical mitotic parthenogen Meloidogyne incognita. Phylogenomic analysis of gene families from M. floridensis, M. incognita and an outgroup species Meloidogyne hapla was carried out to trace the evolutionary history of these species' genomes, and we demonstrate that M. floridensis was one of the parental species in the hybrid origins of M. incognita. Analysis of the M. floridensis genome itself revealed many gene loci present in divergent copies, as they are in M. incognita, indicating that it too had a hybrid origin. The triploid M. incognita is shown to be a complex double-hybrid between M. floridensis and a third, unidentified, parent. The agriculturally important RKN have very complex origins involving the mixing of several parental genomes by hybridization and their extreme polyphagy and success in agricultural environments may be related to this hybridization, producing transgressive variation on which natural selection can act. It is now clear that studying RKN variation via individual marker loci may fail due to the species' convoluted origins, and multi-species population genomics is essential to understand the hybrid diversity and adaptive variation of this important species complex. This comparative genomic analysis provides a compelling example of the importance and complexity of hybridization in generating animal species diversity more generally.


Crystal structure and stacking faults in the layered honeycomb, delafossite-type materials Ag3LiIr2O6 and Ag3LiRu2O6.
Powder samples of Ag3LiIr2O6 and Ag3LiRu2O6 were synthesized from α-Li2IrO3 and Li2RuO3 respectively by ion exchange in an AgNO3 melt. The crystal structures of the title compounds were solved from high resolution laboratory X-ray powder diffraction (XRPD) patterns and from pair distribution function (PDF) analysis using synchrotron X-ray powder diffraction data. In both crystal structures edge sharing LiO6/3- and (Ir/Ru)O6/3-octahedra form honeycomb like layers that are stacked in a staggered fashion. Silver cations, situated in-between the layers mediate the interlayer interactions by linear O-Ag-O bonds. Anisotropic peak broadening in the XRPD patterns and diffuse scattering occurring as streaks in the precession electron diffraction (PED) patterns indicate the presence of stacking faults, which could be also visualized by high resolution transmission electron microscopy (HRTEM). Possible alternative stacking sequences were derived from the ideal crystal and incorporated into a microstructure model. By applying a supercell approach that randomly generates and averages stacking sequences based on transition probabilities and combining it with a grid search algorithm, the microstructures, i.e. the degrees of faulting in the structures of the title compounds were refined to the measured XRPD data. In result the crystal structures of Ag3LiIr2O6 and Ag3LiRu2O6 were found to be vastly faulted with almost no coherence of the stacked layers.


Immunophenotypic signatures of benign and dysplastic granulopoiesis by cytomic profiling.
The role of flow cytometry (FCM) in diagnosing myelodysplastic syndromes (MDS) remains controversial, because analysis of myeloid maturation may involve subjective interpretation of sometimes subtle patterns on multiparameter FCM. Using six-parameter marker combinations known to be useful in evaluating the myeloid compartment in MDS, we measured objective immunophenotypic differences between non-neoplastic (n = 25) and dysplastic (n = 17) granulopoiesis using a novel method, called Fisher information nonparametric embedding (FINE), that measures information distances among FCM datasets modeled as individual high-dimensional probability density functions, rather than as sets of two-dimensional histograms. Information-preserving component analysis (IPCA) was used to create information-optimized "rotated" two-dimensional histograms for visualizing myelopoietic immunophenotypes for each individual sample. There was a consistent trend of segregation of higher-grade MDS (RAEB and RCMD) from benign by FINE analysis. This difference was accentuated in cases with morphologic dysgranulopoiesis and in cases with clonal cytogenetic abnormalities. However, lower grades of MDS or cases that lacked morphologic dysgranulopoiesis showed much greater overlap with non-neoplastic cases. Two cases of reactive left shift were consistently embedded within the higher-grade MDS group. IPCA yielded two-dimensional histogram projections for each individual case by relative weighting of measured cellular characteristics, optimized for preserving information distances derived through FINE. Objective analysis by information geometry supports the conclusions of previous studies that there are immunophenotypic differences in the maturation patterns of benign granulopoiesis and high grade MDS, but also reinforces the known pitfalls of overlap between low-grade MDS and benign granulopoiesis and overlap between reactive granulocytic left shifts and dysplastic granulopoiesis.


Sphingosine kinase 1 regulates pro-inflammatory responses triggered by TNFalpha in primary human monocytes.
Monocytes play an important role in inflammation, angiogenesis, and atherosclerosis. During these processes monocytes release pre-formed proinflammatory mediators from granules, and synthesize de novo cytokines and chemokines important in the amplification of the inflammatory response. One of the most prominent triggers of inflammatory responses is the cytokine TNFalpha. However, the intracellular signaling cascades triggered by TNFalpha are not fully understood. In this study we investigated the roles of SPHK on the TNFalpha-triggered responses on human primary monocytes. We show that TNFalpha rapidly triggers S1P generation and activation of SPHK. Moreover, our data shows that SPHK1 is the isoform activated by TNFalpha, and plays an essential role on the TNFalpha-triggered intracellular Ca2+ signals, degranulation, cytokine production, and activation of NFkappaB, thus suggesting a pivotal role for SPHK1 on the proinflammatory responses triggered by TNFalpha.


Twin pregnancy with fetus papyraceous.
A successful pregnancy outcome with vaginal delivery of a term single live fetus in case of twin pregnancy with other twin papyraceous, (vanished in second trimester) is an uncommon finding. Ten percent of all perinatal deaths are related with dizygotic twins. The rate of intrauterine fetal demise in multiple pregnancies is as high as three times of monozygotic. Most frequently used and suggested method is follow-up of the maternal coagulation system by a series of lab test. Sonography is a visual tool to confirm the diagnosis. In case of monofetal death of twin pregnancies, the type of placenta should be analysed and then follow up and treatment modalities of these cases should be choosen.


A survey of Papua New Guinea's health sector financing and expenditure.
As a result of increasing economic constraints in the health sector of Papua New Guinea, the Department of Health embarked upon a major financial policy review program in 1986, which incorporated several studies in key areas. This paper presents the results of one component of the review, a health sector financing and expenditure study, which was undertaken for the 1986 financial year. The study found that: 1) 88% of health sector expenditure was provided by the public sector; 2) over 95% of expenditure was used for recurrent costs and there was poor utilization of overseas aid for capital projects; 3) the balance of expenditure between primary and secondary health was appropriate; 4) there were regional inequities in expenditure; and 5) a disproportionate amount of recurrent expenditure was consumed by personnel costs. The need to increase nongovernment expenditure on health was identified and further investigation of alternative sources of financing the health sector is required.


Dynamic mechanical finite element model of biological cells for studying cellular pattern formation.
Understanding the geometric, topologic, and mechanical properties of cells and their interactions is critical for studying tissue pattern formation and organ development. Computational model and tools for simulating cell pattern formation have broad implications in studying embryogenesis, blood-vessel development, tissue regeneration, and tumor growth. Although a number of cell modeling methods exist, they do not simultaneously account for detailed cellular shapes as well as dynamic changes in cell geometry and topology. Here we describe a dynamic finite element cell model (dFEMC) for studying populations of cells and tissue development. By incorporating details of cell shape, cell growth and shrinkage, cell birth and death, cell division and fusion, our method can model realistically a variety problems of cell pattern formation. We give two examples of applying our method to the study of cell fusion and cell apoptosis. The dFEMC model developed here provides a general computational framework for studying dynamics pattern formation of tissue.


Glucose-lowering effects and low risk of hypoglycemia in patients with maturity-onset diabetes of the young when treated with a GLP-1 receptor agonist: a double-blind, randomized, crossover trial.
The most common form of maturity-onset diabetes of the young (MODY), hepatocyte nuclear factor 1α (HNF1A diabetes: MODY3) is often treated with sulfonylureas that confer a high risk of hypoglycemia. We evaluated treatment with GLP-1 receptor agonists (GLP-1RAs) in patients with HNF1A diabetes. Sixteen patients with HNF1A diabetes (8 women; mean age 39 years [range 23-67 years]; BMI 24.9 ± 0.5 kg/m(2) [mean ± SEM]; fasting plasma glucose [FPG] 9.9 ± 0.9 mmol/L; HbA1c 6.4 ± 0.2% [47 ± 3 mmol/mol]) received 6 weeks of treatment with a GLP-1RA (liraglutide) and placebo (tablets), as well as a sulfonylurea (glimepiride) and placebo (injections), in randomized order, in a double-blind, crossover trial. Glimepiride was up-titrated once weekly in a treat-to-target manner; liraglutide was up-titrated once weekly to 1.8 mg once daily. At baseline and at the end of each treatment period a standardized liquid meal test was performed, including a 30-min light bicycle test. FPG decreased during the treatment periods (-1.6 ± 0.5 mmol/L liraglutide [P = 0.012] and -2.8 ± 0.7 mmol/L glimepiride [P = 0.003]), with no difference between treatments (P = 0.624). Postprandial plasma glucose (PG) responses (total area under the curve) were lower with both glimepiride (2,136 ± 292 min × mmol/L) and liraglutide (2,624 ± 340 min × mmol/L) compared with baseline (3,127 ± 291 min × mmol/L; P < 0.001, glimepiride; P = 0.017, liraglutide), with no difference between treatments (P = 0.121). Eighteen episodes of hypoglycemia (PG ≤3.9 mmol/L) occurred during glimepiride treatment and one during liraglutide treatment. Six weeks of treatment with glimepiride or liraglutide lowered FPG and postprandial glucose excursions in patients with HNF1A diabetes. The glucose-lowering effect was greater with glimepiride at the expense of a higher risk of exclusively mild hypoglycemia.


Hyperbaric oxygen in the treatment of burns.
Since 1972, over 800 burn victims have been treated with hyperbaric oxygen at the Burn Center of Sherman Oaks Community Hospital in Los Angeles. HBO is used only as an adjuvant to standard resuscitation and is not intended to replace current accepted therapy. The results of the first three-year study program show definite changes in burn care delivery. There has been a definite decrease in the mortality as compared with our own predictions and statistics and with those of the N.B.I.E. In addition, a marked improvement in morbidity and a reduction in hospital stay were noted.


Clinical and economic impacts of latanoprost 0.005% in first-line treatment of open-angle glaucoma and ocular hypertension in France.
To assess the cost-effectiveness of treatment strategies that utilize first-line latanoprost compared to those based on initial beta-blocker therapy in patients with open-angle glaucoma (OAG) or ocular hypertension (OH) in France. The study was based on a decision-analytic model that was populated with data from a retrospective chart review. A hypothetical cohort of patients newly diagnosed with OAG and/or OH was assessed over a period of 2 and 3 years. For each treatment strategy 10,000 patients were assumed. First-line latanoprost therapy was significantly more effective than initial treatment with a beta-blocker, providing more days of intraocular pressure (IOP) control primarily due to its longer time until initial treatment failure. Latanoprost's higher acquisition cost was largely offset by reductions in costs associated with surgical procedures. The additional cost for latanoprost was estimated at approximately 41 Euro and 27 Euro over 2 and 3 years, respectively. The incremental cost per day of IOP control when latanoprost was used as first-line strategy compared to the first-line beta-blocker strategy was 0.82 Euro and 0.36 Euro over 2 and 3 years, respectively. These results provide compelling evidence that first-line latanoprost therapy can provide superior clinical outcomes at a small additional cost in actual clinical practice.


A population-based retrospective cohort study comparing care for Western Australians with and without Alzheimer's disease in the last year of life.
Our study investigated health service use for people in the last year of life, comparing those who died with and without Alzheimer's disease (AD) documented on the death certificate. Using a population-based retrospective cohort design, and utilizing the Western Australian Data Linkage System for the period 2000-2002 (2.5 years), the study found that 992 people died of either AD alone or AD with at least one other condition recorded on the death certificate. Of those with documented AD, 90.4% were aged 75 or more years, two-thirds were female (68.8%), more than one-half were widowed (59.3%) and the majority lived in a major city (77.0%). Most deceased people had comorbidities recorded on death certificates (90.0%); the majority having either two (34.5%) or three (28.8%) comorbid conditions. Over two-thirds of people aged over 75 years with AD died in a residential aged care facility (RACF, 67.4%), while the greatest proportion of people without AD died in hospital (52.9%). When a comparison was made in the use of hospital and community-based services for decedents aged over 75 years with and without AD, dissimilarities were evident. Less than one-half of people with documented AD received hospital care in the last year of life (46.3%) compared to over 80% of people without AD (80.5%). Likewise, fewer people in the Alzheimer's group received community care when compared to those without documented AD (10.8% vs. 28.5%). Despite a small group of people with AD (5.4%) who were transferred to an RACF shortly before dying, most people in this group lived and died in an RACF and had their care provided in this setting. Adequate nursing, medical and allied health services, and the provision of appropriate social support, including the use of advance care directives in RACFs, are essential for equitable provision of care to people with AD.


The content of 90Sr in the bone tissue of the population of the Soviet Union. (1959-1971) (The basic laws of its accumulation and distribution).
On the basis of the results of long-term observations, general laws have been identified determining the dynamics of 90Sr concentration in the bone tissue of the population of the USSR in 1959-71, the peculiarities of 90Sr accumulation in different age groups and the connection and dependences between the intake and the contect of the isotope in human organism. It has been found that maximum 90Sr concentration occur in children born in the year when nuclear tests were carried out. The type of distribution of the frequency of occurrence of cases showing different levels of 90Sr content in uniform population groups was determined. Characteristic features of 90Sr distribution in different bones of the skeleton and its changes in the course of time were demonstrated.


Effects of coronary artery occlusion on thoracic spinal neurons receiving viscerosomatic inputs.
Action potentials of single cells in the gray matter of the T2 to T4 spinal segments of chloralose-anesthetized cats were recorded during occlusion of the anterior descending branch of the left coronary artery. Only cells responding both to visceral and somatic afferent inputs were examined. In 53% of the cells the discharge frequency was unaffected by coronary artery occlusion (CAO). The remaining 47% could be classified by four response patterns: 1) in 18% frequency increased immediately following the onset of CAO; 2) in 22%, frequency increased 13.6 +/- 3.3 (SE) s after CAO and the increased frequency usually was associated with changes in the configuration of the electrocardiogram; 3) in 3%, frequency increased at the onset of CAO and then increased more approximately 11 s later; 4) in 4%, inhibition of spontaneous activity occurred during CAO. A noxious pinch applied to the somatic receptive field maximally excited 94% and inhibited 6% of the cells. Since viscerosomatic convergence provides a basis for referred pain, these cells could potentially be involved with transmission of nociceptive information to regions of the brain that integrate pain sensations.


The more you know: Schema-congruency supports associative encoding of novel compound words. Evidence from event-related potentials.
We aimed to investigate the neurocognitive mechanisms of event congruency with prior (schema) knowledge for the learning of novel compound words. Event-related potentials (ERPs) were recorded during an incidental learning task, in which novel noun-noun compounds were presented in a semantically congruent context, enabling schema-supported processing, or in a neutral context. As expected, associative memory performance was better for compounds preceded by a congruent context. Although the N400 was attenuated in the congruent condition, subsequent memory effects (SMEs) in the N400 time interval did not differ across conditions, suggesting that the processes reflected in the N400 cannot account for the memory advantage in the congruent condition. However, a parietal SME was obtained for compounds preceded by a congruent context, only, which we interpret as reflecting the schema-supported formation of a conceptual compound representation. A late frontal SME was obtained in both conditions, presumably reflecting the more general inter-item associative encoding of compound constituents.


Role of lncRNA NR2F1-AS1 and lncRNA H19 Genes in Hepatocellular Carcinoma and Their Effects on Biological Function of Huh-7.
This research was designed to probe into the expression and related mechanism of lncRNA NR2F1-AS1 and H19 in hepatocellular carcinoma (HCC). Forty-two HCC patients who came to our hospital from February 2018 to August 2019 were included into a research group (RG). Meanwhile, 46 healthy controls were regarded as a control group (CG). BEL-7402, Huh-7 human hepatoma cells and HL-7702 human normal liver cells were purchased, and the NR2F1-AS1 and H19 levels in serum and tissues of HCC patients were detected. PcDNA3.1-NR2F1-AS1, si-NR2F1-AS1, NC, pcDNA3.1-H19 and si-H19 were transfected into BEL-7402 and Huh-7 cells. The NR2F1-AS1 and H19 levels in samples were detected via qRT-PCR, and the expression of apoptosis-related proteins in cells was tested through WB. Cell proliferation, invasion, or apoptosis was detected by CCK8, Transwell or flow cytometry, respectively. The NR2F1-AS1 and H19 levels were high in human hepatoma cells, and AUCs of lncRNA NR2F1-AS1 and lncRNA H19 were both >0.8. The lncRNA NR2F1-AS1 and lncRNA H19 were associated with HCC staging. After transfection of pcDNA3.1-NR2F1-AS1, si-NR2F1-AS1, NC, pcDNA3.1-H19, si-H19 BEL-7402 and Huh-7 cells, silencing NR2F1-AS1 and H19 expression can promote apoptosis and inhibit cell growth, while silencing their over-expression can inhibit the EMT process of Huh-7 cells. lncRNA NR2F1-AS1 and lncRNA H19 genes are abnormally expressed in HCC. Furthermore, the two can suppress the EMT process of Huh-7 cells and promote apoptosis effectively.


Headache in traumatic brain injuries from blunt head trauma.
To determine the risk of traumatic brain injuries (TBIs) in children with headaches after minor blunt head trauma, particularly when the headaches occur without other findings suggestive of TBIs (ie, isolated headaches). This was a secondary analysis of a prospective observational study of children 2 to 18 years with minor blunt head trauma (ie, Glasgow Coma Scale scores of 14-15). Clinicians assessed the history and characteristics of headaches at the time of initial evaluation, and documented findings onto case report forms. Our outcome measures were (1) clinically important TBI (ciTBI) and (2) TBI visible on computed tomography (CT). Of 27 495 eligible patients, 12 675 (46.1%) had headaches. Of the 12 567 patients who had complete data, 2462 (19.6%) had isolated headaches. ciTBIs occurred in 0 of 2462 patients (0%; 95% confidence interval [CI]: 0%-0.1%) in the isolated headache group versus 162 of 10 105 patients (1.6%; 95% CI: 1.4%-1.9%) in the nonisolated headache group (risk difference, 1.6%; 95% CI: 1.3%-1.9%). TBIs on CT occurred in 3 of 456 patients (0.7%; 95% CI: 0.1%-1.9%) in the isolated headache group versus 271 of 6089 patients (4.5%; 95% CI: 3.9%-5.0%) in the nonisolated headache group (risk difference, 3.8%; 95% CI: 2.3%-4.5%). We found no significant independent associations between the risk of ciTBI or TBI on CT with either headache severity or location. ciTBIs are rare and TBIs on CT are very uncommon in children with minor blunt head trauma when headaches are their only sign or symptom.


Inducing Energetic Switching Using Klotho Improves Vascular Smooth Muscle Cell Phenotype.
The cardiovascular disease of atherosclerosis is characterised by aged vascular smooth muscle cells and compromised cell survival. Analysis of human and murine plaques highlights markers of DNA damage such as p53, Ataxia telangiectasia mutated (ATM), and defects in mitochondrial oxidative metabolism as significant observations. The antiageing protein Klotho could prolong VSMC survival in the atherosclerotic plaque and delay the consequences of plaque rupture by improving VSMC phenotype to delay heart attacks and stroke. Comparing wild-type VSMCs from an ApoE model of atherosclerosis with a flox'd Pink1 knockout of inducible mitochondrial dysfunction we show WT Pink1 is essential for normal cell viability, while Klotho mediates energetic switching which may preserve cell survival. Wild-type ApoE VSMCs were screened to identify potential drug candidates that could improve longevity without inducing cytotoxicity. The central regulator of cell metabolism AMP Kinase was used as a readout of energy homeostasis. Functional energetic switching between oxidative and glycolytic metabolism was assessed using XF24 technology. Live cell imaging was then used as a functional readout for the WT drug response, compared with Pink1 (phosphatase-and-tensin-homolog (PTEN)-induced kinase-1) knockout cells. Candidate drugs were assessed to induce pACC, pAMPK, and pLKB1 before selecting Klotho for its improved ability to perform energetic switching. Klotho mediated an inverse dose-dependent effect and was able to switch between oxidative and glycolytic metabolism. Klotho mediated improved glycolytic energetics in wild-type cells which were not present in Pink1 knockout cells that model mitochondrial dysfunction. Klotho improved WT cell survival and migration, increasing proliferation and decreasing necrosis independent of effects on apoptosis. Klotho plays an important role in VSMC energetics which requires Pink1 to mediate energetic switching between oxidative and glycolytic metabolism. Klotho improved VSMC phenotype and, if targeted to the plaque early in the disease, could be a useful strategy to delay the effects of plaque ageing and improve VSMC survival.


Canadian bitumen is engineered for transport, but the type of product produced can affect spill contingency planning.
Canadian bitumen is too viscous to transport by rail and pipeline to markets. One approach to solve this viscosity issue is to dilute the bitumen with a thinning agent to meet transport specifications, but the addition of diluent underutilizes pipeline capacity and increases production cost. A second approach involves the partial refinement of bitumen to produce synthetic crude, which better utilizes pipeline capacity; however, capital and operational costs are high. A third alternative is a new technology that involves wrapping bitumen in a polymer layer to form a solid "puck" termed Canapux, but transportation of this product to coastal ports is limited to rail. Also, greenhouse gas emissions are greater when oil is transported by rail rather than pipeline. In the end, a variety of bitumen products will be transported in Canada, but not all of these products will respond to remediation equally when spilled. In order to ensure effective spill contingency planning, we recommend that engineered bitumen products have physical properties that are resilient to change, within the range of typical response times, after a spill.


Building supply chain capacity for neglected tropical diseases: experience from the Ascend West and Central Africa programme.
The Ascend West and Central Africa programme, funded by the UK Foreign, Commonwealth and Development Office (FCDO) is supporting integrated preventative chemotherapy for up to five neglected tropical diseases (NTDs), including intestinal worms, lymphatic filariasis, river blindness, trachoma and schistosomiasis. The programme is implemented across 13 countries by a consortium of four leading international development partners: Sightsavers, Liverpool School of Tropical Medicine, Schistosomiasis Control Initiative Foundation and Mott Macdonald. This paper presents messages learnt from country assessments that took place prior to the global outbreak of coronavirus disease 2019 (COVID-19). These messages remain relevant post-COVID-19, with greater priority being given to the challenges for national NTD programmes in continuing to deliver mass drug administration (MDA) during the pandemic. Stakeholder coordination from the earliest stages of the pandemic has occurred at two levels: in the first mile with global partners of the NTD Supply Chain Forum and in the last mile with implementing partners in each country. This has been instrumental to manage delayed MDA, including the impact delays have on the shipment of NTD donated drugs and the distribution of stock held in country. The Ascend West and Central Africa programme is supporting countries with the resumption of MDA through a risk assessment and mitigation action (RAMA) process.


Aphids indirectly increase virulence and transmission potential of a monarch butterfly parasite by reducing defensive chemistry of a shared food plant.
Parasites and hosts live in communities consisting of many interacting species, but few studies have examined how communities affect parasite virulence and transmission. We studied a food web consisting of two species of milkweed, two milkweed herbivores (monarch butterfly and oleander aphid) and a monarch butterfly-specific parasite. We found that the presence of aphids increased the virulence and transmission potential of the monarch butterfly's parasite on one milkweed species. These increases were associated with aphid-induced decreases in the defensive chemicals of milkweed plants. Our experiment suggests that aphids can indirectly increase the virulence and transmission potential of monarch butterfly parasites, probably by altering the chemical composition of a shared food plant. These results indicate that species that are far removed from host-parasite interactions can alter such interactions through cascading indirect effects in the food web. As such, indirect effects within ecological communities may drive the dynamics and evolution of parasites.


The plug-unplug sequence: an important step to achieve type II pneumocyte maturation in the fetal lamb model.
The purpose of this study was to test the hypothesis that tracheal obstruction (plugging) in the fetal lamb model leads to a decrease in the absolute number of type II pneumocytes and that reversing the obstruction before birth (unplugging), allows the type II cells to recover while maintaining the beneficial effect on lung growth. Nine time-dated pregnant ewes (term, 145 days), carrying 17 fetuses, were used in this surgical trial. The fetuses were divided into three experimental groups: group A underwent plugging at 93 days gestation, followed by unplugging at 110 days; group B animals had tracheal ligation at 93 days and group C consisted of unoperated controls. All fetuses were delivered by cesarean section at 136 days' gestation. The fetal trachea was obstructed with the tracheoscopically placed detachable balloon described by our group. Unplugging was performed by needle puncture of the balloon under tracheoscopic vision. Outcome measurements consisted of lung-to-body-weight ratio (LWBR), lung morphometry (mean terminal bronchial density [MTBD] and linear intercept [Lm]), and assessment of the number of type II pneumocytes. The latter was determined by in situ hybridization to the mRNA of surfactant protein-C, which is exclusively produced by type II cells. Statistics were calculated using a two-tailed unpaired t test and P less than .05 is considered significant. Seventeen animals are included in the results. All of them had lung samples analyzed for lung morphometry, whereas for type II cells analysis, three animals were studied in each group. Morphometric analyses were consistent with pulmonary hyperplasia for group B, whereas group A lungs showed more histological maturity than group C albeit not as marked as group B. In group A, there was a similar number of type II cells to that observed in group C (53.2 +/- 3.9 v 55.9 +/- 4.0, P = .66). However, for group B animals, the number of type II pneumocytes was markedly decreased compared with controls (4.7 +/- 0.1 v 55.9 +/- 4, P = .0003). The authors conclude that tracheal ligation until birth, although inducing pulmonary hyperplasia, significantly decreases the number of type II pneumocytes in the alveoli. After a temporary 15-day occlusion initiated at 95 days' gestation, there is complete normalization of the density of type II cells. These results bear importance on the duration of PLUG to treat the pulmonary hypoplasia seen in congenital diaphragmatic hernia. Temporary tracheal obstruction now needs to be tested in a hypoplastic lung model.


Variation in the properties of a strain of Staphylococcus aureus isolated over three months from a single hospital.
A strain of Staphylococcus aureus has been isolated from a hospital environment over 3 months. Every isolate was lysed by phage 77, had high-level resistance to streptomycin, and was resistant to about 250 pg per ml of both tetracycline and sulphonamide; a combination of sulphamethoxazole and trimethoprim produced little bacteristatic synergy towards each isolate. All These organisms were thus considered to be "the same"; the variation in other properties was probably due to rapid evolutionary change in vivo. the variation in senxitivity to methicillin and neomycin, and the absence of penicillinase production in some isolates, probably indicated loss of the relevant genes. Several isolates had probably acquired resistance to lincomycin by a one-step mutatuon in vivo. The usefulness of lincomycin and analogues in treating staphylococcal infections seems limited.


Ultrarestrictive Opioid Prescription Protocol for Pain Management After Gynecologic and Abdominal Surgery.
Opioids are routinely prescribed for postoperative home pain management for most patients in the United States, with limited evidence of the amount needed to be dispensed. Opioid-based treatment often adversely affects recovery. Prescribed opioids increase the risk of chronic opioid use, abuse, and diversion and contribute to the current opioid epidemic. To evaluate whether after hospital discharge, postsurgical acute pain can be effectively managed with a markedly reduced number of opioid doses. In this case-control cohort study, an ultrarestrictive opioid prescription protocol (UROPP) was designed and implemented from June 26, 2017, through June 30, 2018, at a single tertiary-care comprehensive cancer center. All patients undergoing gynecologic oncology surgery were included. Patients undergoing ambulatory or minimally invasive surgery (laparoscopic or robotic approach) were not prescribed opioids at discharge unless they required more than 5 doses of oral or intravenous opioids while in the hospital. Patients who underwent a laparotomy were provided a 3-day opioid pain medication supply at discharge. Total number of opioid pain medications prescribed in the 60-day perioperative period, requests for opioid prescription refills, and postoperative pain scores and complications were evaluated. Factors associated with increased postoperative pain, preoperative and postoperative pain scores, inpatient status, prior opioid use, and all opioid prescriptions within the 60-day perioperative window were monitored among the case patients and compared with those from consecutive control patients treated at the center in the 12 months before the UROPP was implemented. Patient demographics and procedure characteristics were not statistically different between the 2 cohorts of women (605 cases: mean [SD] age, 56.3 [14.5] years; 626 controls: mean [SD] age, 55.5 [13.9] years). The mean (SD) number of opioid tablets given at discharge after a laparotomy was 43.6 (17.0) before implementation of the UROPP and 12.1 (8.9) after implementation (P < .001). For patients who underwent laparoscopic or robotic surgery, the mean (SD) number of opioid tablets given at discharge was 38.4 (17.4) before implementation of the UROPP and 1.3 (3.7) after implementation (P < .001). After ambulatory surgery, the mean (SD) number of opioid tablets given at discharge was 13.9 (16.6) before implementation of the UROPP and 0.2 (2.1) after implementation (P < .001). The mean (SD) perioperative oral morphine equivalent dose was reduced to 64.3 (207.2) mg from 339.4 (674.4) mg the year prior for all opioid-naive patients (P < .001). The significant reduction in the number of dispensed opioids was not associated with an increase the number of refill requests (104 patients [16.6%] in the pre-UROPP group vs 100 patients [16.5%] in the post-UROPP group; P = .99), the mean (SD) postoperative visit pain scores (1.1 [2.2] for the post-UROPP group vs 1.4 [2.3] for pre-UROPP group; P = .06), or the number of complications (29 cases [4.8%] in the post-UROPP group vs 42 cases [6.7%] in the pre-UROPP group; P = .15). Implementation of a UROPP was associated with a significant decrease in the overall amount of opioids prescribed to patients after gynecologic and abdominal surgery at the time of discharge for all patients, and for the entire perioperative time for opioid-naive patients without changes in pain scores, complications, or medication refill requests.


[Anxiety-depressive disorders and stroke: possible etiopathogenetic correlations].
Anxiety-depressive disorders (ADD) and stroke are ranked among top ten diseases in the list of the World Health Organization.Comorbid cerebrovascular diseases and ADD worsen the clinical picture, each aggravating the progression and severity of the other. The detailed study of etiopathogenetic correlations between ADD and stroke can assist in the development of the unified pharmacological approach to their treatment.


Asymmetrical flagellar rotation in Borrelia burgdorferi nonchemotactic mutants.
The Lyme disease spirochete Borrelia burgdorferi has bundles of periplasmic flagella subpolarly located at each cell end. These bundles rotate in opposite directions during translational motility. When not translating, they rotate in the same direction, and the cells flex. Here, we present evidence that asymmetrical rotation of the bundles during translation does not depend upon the chemotaxis signal transduction system. The histidine kinase CheA is known to be an essential component in the signaling pathway for bacterial chemotaxis. Mutants of cheA in flagellated bacteria continually rotate their flagella in one direction. B. burgdorferi has two copies of cheA designated cheA1 and cheA2. Both genes were found to be expressed in growing cells. We reasoned that if chemotaxis were essential for asymmetrical rotation of the flagellar bundles, and if the flagellar motors at both cell ends were identical, inactivation of the two cheA genes should result in cells that constantly flex. To test this hypothesis, the signaling pathway was completely blocked by constructing the double mutant cheA1kan cheA2ermC. This double mutant was deficient in chemotaxis. Rather than flexing, it failed to reverse, and it continually translated only in one direction. Video microscopy of mutant cells indicated that both bundles actively rotated. The results indicate that asymmetrical rotation of the flagellar bundles of spirochetes does not depend upon the chemotaxis system but rather upon differences between the two flagellar bundles. We propose that certain factors within the spirochete localize at the flagellar motors at one end of the cell to effect this asymmetry.


The HER2 I655V polymorphism and breast cancer risk in Ashkenazim.
BACKGROUND Over-expression of the human epidermal growth factor receptor 2 (Her2) protooncogene is associated with poor prognosis among female patients with breast cancer. A polymorphism in the HER2 gene (I655V) has been associated with an elevated risk of breast cancer in some ethnic groups. METHODS Subjects from a community-based study of 5318 Ashkenazim from the Washington, DC area were selected for analysis of the I655V HER2 germline polymorphism. We estimated age-specific breast cancer risk from HER2 I655V based on the family history data, using the female first-degree relatives of the study participants and a novel extension of the kin cohort method. RESULTS The estimated cumulative risk of breast cancer to age 70 was approximately 30% higher among HER2 I655V carriers than noncarriers (RR = 1.33; 95% confidence interval [CI] = 1.03-1.83). The effect of the allele seems stronger at younger ages (among women younger than 50 years, RR = 2.11; CI = 1.39-3.28) and especially among younger women with a family history of breast cancer (RR = 8.9; CI = 1.9-19.7). Increased risk of breast cancer associated with the I655V allele was also observed among BRCA1/2 mutation carriers, although these results are based on small numbers. CONCLUSION These analyses suggest that the HER2 valine allele might be associated with increased risk of breast cancer, especially in young women and in women with a family history of the disease.


Radiotelemetered cystometry in pigs: validation and comparison of natural filling versus diuresis cystometry.
Cystometry has previously been performed in awake animals using vesical and abdominal catheters, and artificial bladder filling. Conventional urodynamic recordings may be obtained in this manner, albeit under nonphysiological and potentially stressful conditions. Therefore, we developed a technique to perform continuous, ambulatory cystometric monitoring in pigs. A dual pressure radiotelemetry device was surgically implanted in 22 large white pigs. Vesical and abdominal pressures were recorded and validated, subtracted detrusor pressures were derived and natural fill and diuresis cystometry was compared. Continuous recordings were obtained for 1 to 24 hours, and the devices remained in the animals for up to 3 months. There were few complications and incrustation of the intravesical catheter tip occurred but it did not appear to affect recorded pressures. The pressure data were validated by comparison with filling pressures during bladder distention and simultaneous conventional cystometry at the end of the experimental period. Comparison of natural filling and diuresis cystometrograms showed that natural bladder filling results in higher maximum detrusor pressure during voiding (38.1 versus 33.9 cm. H2O, p <0.05), higher detrusor pressure after contractions (42.6 versus 32.2 cm. H2O, p <0.05) and more frequent detection of unstable contractions in pigs with detrusor instability secondary to experimental manipulation of the lower urinary tract (77.8% versus 45.0%, p <0.05). This technique allows continuous cystometric monitoring in less stressed animals under more physiological conditions for relatively long periods and, thus, allows prolonged assessment of bladder function in pigs in response to pathological and pharmacological manipulations. Nonphysiological rates of bladder filling have been shown to result in detrusor inhibition, which emphasizes the importance of ambulatory cystometry when describing bladder function.


Evaluation of the toxicity of 17 essential oils against Choristoneura rosaceana (Lepidoptera: Tortricidae) and Trichoplusia ni (Lepidoptera: Noctuidae).
The obliquebanded leafroller, Choristoneura rosaceana Harris, and the cabbage looper, Trichoplusia ni Hübner, are serious fruit and vegetable pests requiring multiple insecticide applications per year. To reduce non-target exposure to hazardous insecticides and to curb resistance development, alternative controls are required. Accordingly, a selection of 17 essential oils was screened against both lepidopteran pests, and the influence of azinphos-methyl resistance on essential oil toxicity to C. rosaceana was studied. Of the 17 essential oils screened, patchouli oil (Pogostemon cablin Benth.) and thyme oil (Thymus vulgaris L.) were selected for further testing against C. rosaceana, whereas patchouli oil, garlic oil (Allium sativum L.) and lemongrass oil (Cymopogon nardus L.) were selected for further testing against T. ni. LC(50) and LD(50) values confirmed that patchouli oil was the most toxic to C. rosaceana larvae, with LC(50) = 2.8 µL mL(-1) and LD(50) = 8.0 µg insect(-1). Garlic oil was the most toxic oil to T. ni larvae with LC(50) = 3.3 µL mL(-1) and LD(50) = 22.7 µg insect(-1), followed by patchouli oil and lemongrass oil. Azinphos-methyl-resistant leafrollers were 1.5-fold more tolerant to patchouli oil and 2.0-fold more tolerant to thyme oil. Based on these results, patchouli oil and other essential oils have sufficient efficacy to be considered as components of an essential oil-based insecticide that targets these lepidopteran pests.


Use of Simulation to Test Systems and Prepare Staff for a New Hospital Transition.
To describe the development and key outcomes arising from the use of simulation as a method to test systems and prepare staff for a transition to a new hospital. We describe a simulation program developed by key parties with the goal of reducing latent safety threats present at the opening of a new hospital and to train staff in new workflows. Issues identified were collected and reported to leadership. Outcomes included the number of learners reached, issues identified (grouped by theme), and results of a postmove survey of hospital-based staff. Approximately 258 hours of simulation were conducted, impacting 514 participants. We conducted 64 hours of system testing and 196 hours of training during the main orientation process. Approximately 641 unique issues were identified (175 equipment, 136 code alarm, 174 barriers to care, and 156 incorrect signage). In a hospital-wide survey, 38% reported simulation as part of their training (39% of nurses and 23% of physicians). 43% of survey respondents reported multidisciplinary simulations; 55% of simulation attendees felt that the simulation was helpful and eased their transition to the new hospital. Systems testing and education using simulation can play a meaningful role in new facility training. Key lessons included early planning, allocation of resources to the effort, flexibility to adapt to changes, and planned integration with other training activities. A formal a priori plan to address issues identified during the process is necessary.


The actual insulation of multilayer clothing.
The effect of geometric factors on insulation was calculated mathematically for standing humans. It was found that internal radiative heat transfer in an ensemble was significant for insulation, that intrinsic clothing insulation is a useful concept only for indoor climates, and that shape plays a minor role. The literature agrees closely on insulation and clothing surface area figures, and the latter are compatible with model predictions. Finally, it was shown that wind, body motion, the effects of posture, and the fit of garments are predictable. Sitting provides more insulation than standing for light clothing, but the reverse is true for heavy clothing. Insulation is decreased by about 20% by cycling and by about 40% by walking, and a reasonable estimate can be made of the effect of wind and wind and motion together. The effect of air motion on vapor permeability is stronger than the effect on heat transfer.


Percutaneous transluminal angioplasty for intracranial atherosclerotic lesions: evolution of technique and short-term results.
A retrospective analysis of a 9-year experience with balloon angioplasty for intracranial atherosclerotic stenosis was undertaken with the goals of illustrating development of a safe technique for treatment of intracranial atherosclerotic disease and reporting the immediate results in this series of patients. Three distinct periods are defined, based on the technique used. In the early period, in which eight patients were treated, the angioplasty balloon size approximated the vessel size, but was always smaller. Angioplasty was moderately rapid and brief (15-30 seconds). Clinical improvement occurred in seven (87.5%) of eight patients, dissection without consequence occurred in four (50%) of eight, and residual stenosis greater than 50% was found in three (37.5%) of eight. No neurological complications occurred. In the middle period, in which 12 patients were treated, the balloon size approximated the vessel size, but oversizing by up to 0.25 mm was permitted. Angioplasty was extremely rapid and brief. Angiographically visible dissection occurred in nine (75%) of 12 patients, necessitating urokinase infusion in five (41.7%) of 12 and producing abrupt occlusion in one (8.3%) of 12, resulting in death. Occlusion secondary to the recrossing of the lesion occurred in one (8.3%) of 12, resulting in stroke. Good outcome was eventually achieved in 10 (83.3%) of 12. In the current period, in which 50 patients have been treated, the balloon is always undersized and inflation is extremely slow (several minutes). Dissection occurred in seven (14%) of 50 patients, necessitating fibrinolysis in two of 50 (4%, both uneventful) and producing no abrupt occlusion or stroke. Residual stenosis greater than 50% occurred in eight (16%) of 50, with no stenosis greater than 70%. Late restenosis occurred in four (9%) of 44 and successful repeated angioplasty was performed in all four. One guidewire vessel perforation occurred (2%), resulting in the patient's death. Good angiographic and short-term clinical outcome was achieved in the other 49 patients (98%). Extremely slow balloon inflation combined with balloon undersizing results in decreased intimal damage, decreased acute platelet/thrombus deposition, and decreased acute closure. This technique sometimes yields suboptimal angiographic results but achieves the clinical goal safely. Intracranial angioplasty can be safely performed using this technique and modern equipment.


1 alpha, 25-Dihydroxyvitamin D3 nuclear receptors in pituitary.
Specific binding of 1 alpha,25-dihydroxyvitamin D(3) was found in nuclear and cytosol fractions of the bovine pituitary. For nuclear binding. the dissociation constant was 0.1 namomole per liter, and maximum binding was 104 femtomoles per milligram of protein. In competition studies, 25-hydroxyvitamin D(3) was 300 times weaker than 1 alpha,25-dihydroxyvitamin D(3). The existence of high-affinity sites supports a physiologic role for 1 alpha,25-dihydroxyvitamin D(3) in the pituitary.


Relationship between lung function and physical performance in disabled older women.
Understanding the association between lung function and physical performance in disabled older women helps in determining the potential for prevention and treatment strategies to decrease disability. The aim of this study was to determine the association of lung function with objective and self-reported physical performance in community-dwelling disabled older women. The Women's Health and Aging Study I consists of 1002 disabled community-dwelling women aged > or = 65. Of these women, 840 underwent spirometry with determination of forced expiratory volume in one second (FEV1) and forced vital capacity (FVC). Cross-sectional analyses included multivariate linear regression to assess the association between FEV1, FVC, FEV1/FVC, and the time to walk four meters after adjusting for confounders, including age, race, geriatric depression scale score, body mass index, muscle strength, osteoarthritis, smoking status, and cardiovascular disease. Multiple logistic regression was used to assess the association between FEV1, FVC, FEV1/FVC, and self-reported disability in physical performance. FEV1 was independently associated with time to walk 4 meters. For every 100 ml decrease in FEV1, there was a 0.15-second (95% confidence interval: 0.24 to 0.06) increase in time to walk 4 meters. There was an 8% increase in the prevalent odds of self-reported disability in physical performance for every 100 ml decrease in FEV1. FVC was also associated with physical performance measures. In contrast, FEV1/FVC was associated with objective but not subjective physical performance. Decreasing lung function is independently associated with decrements in objective and self-reported physical performance in disabled older women.


Social learning of acoustic anti-predator cues occurs between wild bird species.
In many species, individuals gather information about their environment both through direct experience and through information obtained from others. Social learning, or the acquisition of information from others, can occur both within and between species and may facilitate the rapid spread of antipredator behaviour. Within birds, acoustic signals are frequently used to alert others to the presence of predators, and individuals can quickly learn to associate novel acoustic cues with predation risk. However, few studies have addressed whether such learning occurs only though direct experience or whether it has a social component, nor whether such learning can occur between species. We investigate these questions in two sympatric species of Parids: blue tits (Cyanistes caeruleus) and great tits (Parus major). Using playbacks of unfamiliar bird vocalizations paired with a predator model in a controlled aviary setting, we find that blue tits can learn to associate a novel sound with predation risk via direct experience, and that antipredator response to the sound can be socially transmitted to heterospecific observers, despite lack of first-hand experience. Our results suggest that social learning of acoustic cues can occur between species. Such interspecific social information transmission may help to mediate the formation of mixed-species aggregations.


Frontal cortex, laterality, and memory: encoding versus retrieval.
The cerebral hemispheres differ in their capabilities and response to verbal versus nonverbal visual material. A priori, it might thus be expected that the right hemisphere would be best activated during a mnemonic task with fMRI when using nonverbalizable images, and the left hemisphere with verbal material. However, previous psychological tests had shown a high degree of similarity in measures of memory for these disparate items. It was thus hypothesized that extensive commonality in the areas activated would prevail when this previously tested material was employed with fMRI. Six subjects underwent fMRI with four types of trials in blocks: fixating; passively viewing 12 words and 12 nonverbalizable images; endeavoring to remember (encoding) another set of 12 words and images; endeavoring to recognize (retrieve) previously viewed words or images. Passive viewing produced small islands of activation in left versus right frontal cortex for words and images, respectively. Endeavoring to remember enlarged the areas of activation and produced some bilaterality. Retrieval greatly augmented activation as well as bilaterality, and some 20% of the activated frontal volume was shared by words and images. Thus, on the one hand, the distribution of activation upon retrieval differed substantially for words versus images, but on the other, as predicted, there was considerable commonality. Predominant laterality of activation in some areas shifted between encoding and retrieval (HERA), importantly involving different regions for words versus images. Of course, processes other than memory per se are undoubtedly involved in these distributions of fMRI activation in frontal cortex, yet the nature of the to-be-remembered items is clearly a major factor, in accord with the asymmetric lateralization in their basic representation.


Spirituality factors in the prediction of outcomes of PTSD treatment for U.S. military veterans.
Spirituality is a multifaceted construct that might affect veterans' recovery from posttraumatic stress disorder (PTSD) in adaptive and maladaptive ways. Using a cross-lagged panel design, this study examined longitudinal associations between spirituality and PTSD symptom severity among 532 U.S. veterans in a residential treatment program for combat-related PTSD. Results indicated that spirituality factors at the start of treatment were uniquely predictive of PTSD symptom severity at discharge, when accounting for combat exposure and both synchronous and autoregressive associations between the study variables, βs = .10 to .16. Specifically, veterans who scored higher on adaptive dimensions of spirituality (daily spiritual experiences, forgiveness, spiritual practices, positive religious coping, and organizational religiousness) at intake fared significantly better in this program. In addition, possible spiritual struggles (operationalized as negative religious coping) at baseline were predictive of poorer PTSD outcomes, β = .11. In contrast to these results, PTSD symptomatology at baseline did not predict any of the spirituality variables at posttreatment. In keeping with a spiritually integrative approach to treating combat-related PTSD, these results suggest that understanding the possible spiritual context of veterans' trauma-related concerns might add prognostic value and equip clinicians to alleviate PTSD symptomatology among those veterans who possess spiritual resources or are somehow struggling in this domain.


[Spontaneous subdural hematoma in a 18-day-old male newborn infant with severe hemophilia A].
Most intracranial bleedings in hemophiliacs occur in patients more than 6 months of age. In the neonatal period, this complication is rare and almost always observed in the first week of life. Based on a review of the literature, intracranial hemorrhage is an exceedingly rare occurrence in infants with hemophilia aged 2 weeks-6 months. We report on a male infant with hemophilia A who was referred to our hospital on day 18 because of pallor and jaundice. The neurological examination was normal. A cerebral ultrasound showed a left sided subdural hematoma with a shift of the midline structures to the right. Packed red blood cells (10 ml/kg) and factor VIII replacement (250 IU) were rapidly instituted and a craniotomy with evacuation of the hematoma was performed. The postoperative course was uneventful. On discharge, the neurological examination was considered normal.


[Counseling regarding healthy lifestyles in primary healthcare and the dietary practices of clients].
This cross-sectional study examines a counseling program on healthy lifestyles run by health care professionals to establish the adoption of healthy dietary practices by patients attending a primary health care unit. Participants in the study included 417 clients of the unit, the majority of whom were women (78.9%), with an average age of 39 years, a high incidence of excessive weight, (59.1%), important dietary inadequacies, and with a contrastingly low frequency of receiving counseling (40.8%). Clients receiving counseling displayed more appropriate consumption of candy/gum (p=0.031), soft drinks (p=0.036), salty foods (p=0.037), artificial flavorings (p=0.005) and eggs (p=0.010). Adoption of healthy dietary practices was more common among older individuals and women (p <0.05). Despite the importance of nutritional counseling in dealing with such health problems, this was not prevalent, suggesting the need for greater intervention by health care professionals aimed at preventing and controlling disease and promoting good health.


Evolutionary history of redox metal-binding domains across the tree of life.
Oxidoreductases mediate electron transfer (i.e., redox) reactions across the tree of life and ultimately facilitate the biologically driven fluxes of hydrogen, carbon, nitrogen, oxygen, and sulfur on Earth. The core enzymes responsible for these reactions are ancient, often small in size, and highly diverse in amino acid sequence, and many require specific transition metals in their active sites. Here we reconstruct the evolution of metal-binding domains in extant oxidoreductases using a flexible network approach and permissive profile alignments based on available microbial genome data. Our results suggest there were at least 10 independent origins of redox domain families. However, we also identified multiple ancient connections between Fe2S2- (adrenodoxin-like) and heme- (cytochrome c) binding domains. Our results suggest that these two iron-containing redox families had a single common ancestor that underwent duplication and divergence. The iron-containing protein family constitutes ∼50% of all metal-containing oxidoreductases and potentially catalyzed redox reactions in the Archean oceans. Heme-binding domains seem to be derived via modular evolutionary processes that ultimately form the backbone of redox reactions in both anaerobic and aerobic respiration and photosynthesis. The empirically discovered network allows us to peer into the ancient history of microbial metabolism on our planet.


Phorbol ester increases mitochondrial cholesterol content in NCI H295R cells.
The first step in steroidogenesis is cholesterol mobilization from cytosolic lipid droplets to the initiating rate-limiting enzyme complex located on the inner mitochondrial membrane. Angiotensin II (AngII), the primary agonist of aldosterone secretion from adrenal glomerulosa cells, is known to induce cholesterol mobilization to mitochondria. However, the role of the protein kinase C (PKC) pathway in mediating cholesterol mobilization is unknown. To determine PKC's involvement, human adrenocortical carcinoma cells were incubated with or without PKC-activating phorbol 12-myristate 13-acetate (PMA) and mitochondrial cholesterol content assayed. Like AngII, PMA significantly elevated mitochondrial cholesterol content as well as aldosterone secretion. Thus, PKC may play a role in cholesterol mobilization to mitochondria and hence steroid production. Atrial natriuretic peptide (ANP) inhibited both AngII- and PMA-stimulated mitochondrial cholesterol content. These findings suggest that the ability of ANP to inhibit steroidogenesis induced by multiple agents may be related to its capacity to reduce cholesterol mobilization.


Treg-Centric View of Immunosuppressive Drugs in Transplantation: A Balancing Act.
Regulatory CD4+ Foxp3+ T cells (Tregs) are critical in controlling immunity and tolerance. Thus, preserving Treg numbers and function in transplanted patients is essential for the successful minimization of maintenance immunosuppression. Multiple cellular signals control the development, differentiation, and function of Tregs. Many of these signals are shared with conventional Foxp3- T cells (Tconv) and are targeted by immunosuppressive drugs, negatively affecting both Tregs and Tconv. Because intracellular signals vary in optimal intensity in different T cell subsets, improved specificity in immunosuppressive regimens must occur to benefit long-term transplant outcomes. In this regard, recent advances are gradually uncovering differences in the signals required in Tregs and Tconv biology, opening the door to new potential therapeutic approaches to either enhance or spare Tregs. In this review, we will explain the prominent cell signaling pathways critical for Treg maintenance and function, while reporting the effects of immunosuppressive drugs targeting these signaling pathways in clinical transplantation settings.


[Immunity status of guinea pigs infected with Brucella L forms].
B. abortus L-forms injected subcutaneously into guinea pigs adapt in the lymph nodes of the animals in the absence of reversion to normal cells. Complete and incomplete antibodies belonging to macro- and microglobulins (IgM and IgG) were synthetized. The allergic transformation of the organism is faintly pronounced. After this form of infection guinea pigs become resistant to B. melitensis infection for 6 months (the term of observation).


The role of nitric oxide in oocyte meiotic maturation and ovulation: meiotic abnormalities of endothelial nitric oxide synthase knock-out mouse oocytes.
Evidence supports the involvement of nitric oxide (NO) in ovulation, steroidogenesis, and atresia-related apoptosis. This study was designed to investigate the role of endothelial nitric oxide synthase (eNOS)-derived NO in ovulation, oocyte meiotic maturation, and ovarian steroidogenesis using wild-type (WT) mice and mice in which the gene for eNOS had been deleted (eNOS knock-out). We observed that mature eNOS knock-out females have significantly fewer pups born in each litter and a higher mortality rate of pups than those born to heterozygote or WT females (P < 0.05). To determine the influence of eNOS deficiency on ovarian function, immature WT and eNOS knock-out mice were superovulated by injecting PMSG (5 IU) followed by an injection of hCG (5 IU, i.p.) 48 h later. To determine whether murine oocytes expressed eNOS before (0 and 8 h post-hCG) and after ovulation (16 h post-hCG). we performed immunofluorescent staining. Positive specific staining for eNOS was observed on the surface of ovarian and ovulated oocytes obtained from WT mice, but not on oocytes from eNOS knock-out mice. To determine the role of eNOS-derived NO in ovulation, ovulated oocytes were counted 16 h post-hCG. eNOS knock-out females showed a significant reduction (by 63%; P < 0.0001) in ovulatory efficiency compared with WT females. The reduction in the ovulation rate in eNOS-deficient mice compared with that in WT mice was also associated with a higher concentration of estradiol (P < 0.01) without significant changes in the plasma progesterone level. eNOS deficiency impaired not only ovulation, but also oocyte meiotic maturation. Ovulated oocytes were classified as being in one of the following stages of meiosis: metaphase I, metaphase II, or showing atypical (degenerative) morphology. We observed that fewer oocytes from eNOS knock-out mice had entered metaphase II of meiosis, and a greater percentage remained in metaphase I or were atypical (P < 0.002) relative to those in WT mice. Furthermore, many oocytes that showed either a delay in meiotic maturation or abnormal morphology were undergoing cell death. Our results support a role for NO in the ovulatory process. The ovarian defects observed in the eNOS knock-out mice suggest that eNOS-derived NO is a modulator of oocyte meiotic maturation.


Psychiatric emergencies in a tertiary care hospital.
People with psychiatric problems often visit a general hospital. Many of them require emergency service. B P Koirala Institute of Health Sciences (BPKIHS) is one of the few health institutes in Nepal providing comprehensive 24-hour psychiatric emergency service. This study aims to document the pattern of psychiatric emergencies in a tertiary care hospital of Eastern Nepal. All psychiatric emergencies referred from different departments in 30 random days during a six-month period were enrolled. This is a descriptive study with convenience sampling method where the diagnoses were made based on the'International Classification of Disease and Infirmity' (ICD-10). One hundred twenty-nine cases were referred in the study period. The M:F ratio was 1.3:1. The service users were predominantly young adults. The majority of cases were seen in the emergency department and medical ward. The most common causes for the consultation were behavioral problems (39%), altered consciousness (32%) and somatic complaints (17%). Approximately 83% received the diagnosis of Category F of the ICD-10. Mental and behavioral disorder due to substance use (F10-19) was the most common disorder (30%), followed by mood/affective disorders (23%) and neurotic, stress-related anxiety disorders (16%). Roughly 46% had co-morbid physical illnesses and 8% received only a physical diagnosis. About 20% had attempted suicide using different means, poisoning being the most common. Emergency psychiatric consultation for mental problems is sought by almost all departments of a general hospital. These consultations are most commonly sought for substance use, mood or anxiety disorders.


Human NH2 terminal of pro-opiomelanocortin as a potential marker for pulmonary carcinoma.
Levels of immunoreactive (IR) human NH2 terminal (HNT) of pro-opiomelanocortin were determined serially in 80 patients with various pulmonary carcinomas, 20 patients with various pulmonary diseases without proven carcinoma, and 32 normal control subjects. Sixty +/- 14% (S.E.) (95% confidence interval) of the patients with proven pulmonary carcinoma had elevated values of IR HNT in plasma (greater than 100 pg/ml); the highest frequency of elevated IR HNT was found in patients with small-cell carcinoma [74 +/- 14.5% (95% confidence interval)]. In eight patients with pulmonary tumor who underwent surgical thoracic exploration, the plasma level of IR HNT in the pulmonary vein was 50.6 +/- 21.4% (95% confidence interval; N = 8) higher than that in the pulmonary artery. The levels of IR HNT correlated well with those of plasma adrenocorticotropic hormone (IR ACTH) (r = 0.988; p less than 0.001); however, the level of IR HNT was 3- to 4-fold higher than that of IR ACTH. Gel permeation chromatography of plasma of patients with pulmonary carcinoma revealed a major peak of IR HNT coinciding with the elution volumes of authentic HNT (Mr 12,000) and a minor peak of IR HNT near the void volumes. These results suggest that HNT originating from pulmonary carcinoma is chemically and immunologically similar to that from human pituitary gland and that measurement of plasma IR HNT may indicate the state of secretion and/or metabolism of peptides related to the expression of the pro-opiomelanocortin gene in human pulmonary carcinoma. The HNT immunoassay is a simple procedure which, contrary to that of ACTH, does not require plasma extraction prior to the assay. This suggests the possibility that measurement of IR HNT alone and in conjunction with other biochemical markers could be a useful parameter for diagnosis and follow-up of patients with pulmonary carcinoma.


Surgical and Non-Surgical Interventions in Complete Rotator Cuff Tears.
This systematic review compares the efficacy and safety of surgical and non-surgical interventions for full- thickness rotator cuff tears. A systematic literature search was conducted in five databases. Randomized (RCTs) and non-randomized controlled trials of interventions (non-RCTs) for the surgical or non-surgical treatment of patients with traumatic or atraumatic full-thickness rotator cuff tears were included. The review protocol was published in the PROSPERO registry (CRD42018100343). Ten studies (three RCTs with 332 participants; seven non-RCTs with 650 participants) met the inclusion criteria. One year after treatment, shoulder function, measured with the 100-point Constant score, had improved by 6.7 points (95% confidence interval [2.3; 11.0]) and pain, measured with the 10-cm visual analog scale, by 1.1 cm (0.5; 1.7] in the full-thickness rotator cuff tears treated surgically compared with non-surgical treatment. In one study the difference in favor of surgery persisted after 10 years' follow-up. For other outcomes, such as range of motion, muscle strength, quality of life, and adverse events, the data were sparse and the group differences were similar. The findings of the non-RCTs were comparable with those of the RCTs. With regard to functional improvement and pain reduction, surgical treatment of full-thickness rotator cuff tears was superior to non-surgical treatment in the short and the long term. Whether the differences between the groups are relevant for individual cases is uncertain, as the measured results were distributed below and above the threshold of clinical relevance. The conclusions may not be applicable to rotator cuff tears over 3 cm in size or to young persons.


School-based prenatal services: can similar outcomes be attained in a nonschool setting?
The purpose of this study is to evaluate whether enhancement of hospital-based prenatal care of adolescents results in pregnancy outcomes comparable to those found in adolescents receiving care at school-based clinics. An initial study comparing hospital clinic and school clinic programs administered by the St. Paul Maternal and Infant Care Project (MIC) from 1973-1976 indicated that delivered teens from the high school clinics had earlier and more frequent prenatal visits and fewer low birth weight babies than delivered adolescents who received care at hospital based clinics. After the initial study of 1976, MIC hospital based services for the adolescents were enhanced to include additional educational and support services. A follow up study, (1976-1979) was subsequently conducted, using criteria similar to the previous study, to compare the results of hospital and school-based programs for pregnant teens. The follow up data demonstrated that the School Group initiated care much earlier and had significantly more prenatal visits than the Comparison Group, but the Comparison Group demonstrated a dramatic improvement in both areas when compared to the first study. Rates of obstetrical complications and infant outcomes were more similar for both groups than in the initial study, supporting the premise that while the school provides a superior setting for provision of prenatal services, similar services at nonschool sites can be greatly enhanced and can demonstrate significant improvement in obstetrical outcomes.


Trolox, a water-soluble analogue of α-tocopherol, photoprotects the surface-exposed regions of the photosystem II reaction center in vitro. Is this physiologically relevant?
Can Trolox, a water-soluble analogue of α-tocopherol and a scavenger of singlet oxygen ((1)O(2)), provide photoprotection, under high irradiance, to the isolated photosystem II (PSII) reaction center (RC)? To answer the question, we studied the endogenous production of (1)O(2) in preparations of the five-chlorophyll PSII RC (RC5) containing only one β-carotene molecule. The temporal profile of (1)O(2) emission at 1270 nm photogenerated by RC5 in D(2)O followed the expected biexponential behavior, with a rise time, unaffected by Trolox, of 13 ± 1 μs and decay times of 54 ± 2 μs (without Trolox) and 38 ± 2 μs (in the presence of 25 μM Trolox). The ratio between the total (k(t)) and chemical (k(r)) bimolecular rate constants for the scavenging of (1)O(2) by Trolox in aqueous buffer was calculated to be ~1.3, with a k(t) of (2.4 ± 0.2) × 10(8) M(-1) s(-1) and a k(r) of (1.8 ± 0.2) × 10(8) M(-1) s(-1), indicating that most of the (1)O(2) photosensitized by methylene blue chemically reacts with Trolox in the assay buffer. The photoinduced oxygen consumption in the oxygen electrode, when RC5 and Trolox were mixed, revealed that Trolox was a better (1)O(2) scavenger than histidine and furfuryl alcohol at low concentrations (i.e., <1 mM). After its incorporation into detergent micelles in unbuffered solutions, Trolox was able to photoprotect the surface-exposed regions of the D1-D2 heterodimer, but not the RC5 pigments, which were oxidized, together with the membrane region of the protein matrix of the PSII RC, by (1)O(2). These results are discussed and compared with those of studies dealing with the physiological role of tocopherol molecules as a (1)O(2) scavenger in thylakoid membranes of photosynthetic organisms.


Analgesic effects and arthritic changes following tramadol administration in a rat hip osteoarthritis model.
We investigated the analgesic effects of tramadol and the arthritic changes following tramadol administration in the rat hip osteoarthritis (OA) model using mono-iodoacetate (MIA). The right hip joints of male Sprague-Dawley rats (n = 5 rats/group) in the Sham group were injected with 25 μl of sterile saline and 1% of fluorogold (FG) retrograde neurotracer. In the MIA + Vehicle and MIA + Tramadol groups, FG and 25 μl of sterile saline with 0.5 mg of MIA were injected into the right hip joint. The MIA + Vehicle and MIA + Tramadol groups were administered daily for 4 weeks, either sterile saline (10 mg/kg, intraperitoneal [i.p.]) or tramadol (10 mg/kg, i.p.). We assessed hyperalgesia every week after MIA administration. Histopathological changes and immunoreactive neurons for calcitonin gene-related peptide (CGRP) in dorsal root ganglia (DRG) were evaluated after 4 weeks of treatment. MIA injection into the hip joint led to mechanical hyperalgesia (p < 0.01), which was significantly reduced by tramadol administration (p < 0.01). Furthermore, daily i.p injection of tramadol significantly suppressed CGRP expression in DRG (p < 0.0001). MIA + Vehicle and MIA + Tramadol groups showed significant cartilage reduction and degeneration compared to the Sham group (p < 0.0001). Interestingly, OA changes significantly progressed in the MIA + Tramadol group compared to the MIA + Vehicle group (p < 0.0001).


Return of shoulder girdle function after anterior versus posterior adolescent idiopathic scoliosis surgery.
Multicenter, prospective, cohort study. To compare minimum 2-year postoperative shoulder function after 3 different surgical approaches for the treatment of primary thoracic curves in patients with adolescent idiopathic scoliosis (AIS). Thoracic spinal instrumentation and fusion can be performed via posterior (PSF), open anterior (OASF), or thoracoscopic anterior (TASF) techniques. Although the morbidity of these 3 surgical approaches is beginning to be understood, no reports have been published comparing 2-year postoperative shoulder strength and range of motion. AIS patients who underwent selective fusion of primary thoracic curves were studied. Right-sided shoulder strength (flexion, abduction) and range of motion (flexion, extension, abduction) measurements were collected prospectively at selected intervals. An arbitrary threshold (80% of preoperative value) was defined as postoperative return of normal shoulder function. Univariate analysis of variance (P < 0.05) was used to compare differences in shoulder function for the 3 approaches at each postoperative time-point. Ninety-two patients with minimum 2-year postoperative shoulder function data were included in this study (24 PSF, 32 OASF, and 36 TASF). On average, patients who had an OASF failed to reach the 80% threshold for right shoulder forward flexion and abduction strength until 1-year and 6-months after surgery, respectively; whereas patients that had a TASF or PSF returned to normal shoulder strength by the 3-month follow-up visit. With regards to ROM, patients in all 3 groups surpassed the 80% preoperative threshold for right shoulder active forward flexion and extension by the 6-week visit, with no clinically significant differences between the groups. However, for active abduction range of motion, patients that had an OASF required 3-months to regain 80% of their preoperative motion, compared to 6-weeks for patients in both the TASF and PSF groups. Approach-related differences in shoulder morbidity do exist in the treatment of primary thoracic curves in AIS. Specifically, OASF imparts a significantly greater magnitude and duration of postoperative shoulder dysfunction than do the TASF or PSF approaches. Nonetheless, these negative effects are transient as shoulder function in those patients treated via an open thoracotomy normalized by the 1-year postoperative time-point at the latest. Accordingly, viewed in isolation, shoulder morbidity should not deter surgeons from using an open anterior approach in the surgical treatment of AIS.


Management of Multiple Sclerosis Symptoms and Comorbidities.
This article discusses the prevalence, identification, and management of multiple sclerosis (MS)-related symptoms and associated comorbidities, including complications that can present at all stages of the disease course. The impact of comorbidities on the outcome of MS is increasingly recognized. This presents an opportunity to impact the course and outcome of MS by identifying and treating associated comorbidities that may be more amenable to treatment than the underlying inflammatory and neurodegenerative disease. The identification of MS-related symptoms and comorbidities is facilitated by brief screening tools, ideally completed by the patient and automatically entered into the patient record, with therapeutic suggestions for the provider. The development of free, open-source screening tools that can be integrated with electronic health records provides opportunities to identify and treat MS-related symptoms and comorbidities at an early stage. Identification and management of MS-related symptoms and comorbidities can lead to improved outcomes, improved quality of life, and reduced disease activity. The use of brief patient-reported screening tools at or before the point of care can facilitate identification of symptoms and comorbidities that may be amenable to intervention.


Factors predictive of subsequent injury in a longitudinal cohort study.
The aims are to: (1) Determine the cumulative incidence of self-reported subsequent injury (of any anatomical site or injury type) occurring between 3 months and 12 months after a sentinel injury among participants in the Prospective Outcomes of Injury Study in New Zealand and (2) Examine the preinjury and injury-related predictors of subsequent injury. Prospective Outcomes of Injury Study participants (n=2282) were interviewed 3 months, 5 months and 12 months after a sentinel injury event. Data were collected about a range of preinjury and injury-related factors at the 3-month interview and about subsequent injury at the 5-month and 12-month interviews. Poisson regression modelling was used to determine the preinjury and injury-related predictors of subsequent injury. Between the 3-month and 12 month interviews 28% of the participants reported at least one subsequent injury. Subsequent injury was 34% more likely among participants with a prior injury affecting them at the time of the sentinel injury compared with participants without a prior injury affecting them, and more likely among non-workers (31% more likely) and trade/manual workers (32% more likely) compared with professionals. Participants whose sentinel injury was due to assault were 43% more likely to report a subsequent injury compared with those whose sentinel injury was accidental. A subsequent injury was 23% less likely if the sentinel injury was a lower extremity fracture compared with other injuries, and 21% less likely if the sentinel injury event involved hospitalisation. Among general injury populations it may be possible to identify people at increased risk for subsequent injury.


[Medical borderlands: engineering the body with plastic surgery and hormonal therapies in Brazil].
This paper explores medical borderlands where health and enhancement practices are entangled. It draws on fieldwork carried out in the context of two distinct research projects in Brazil on plastic surgery and sex hormone therapies. These two therapies have significant clinical overlap. Both are made available in private and public healthcare in ways that reveal the class dynamics underlying Brazilian medicine. They also have an important experimental dimension rooted in Brazil's regulatory context and societal expectations placed on medicine as a means for managing women's reproductive and sexual health. Off-label and experimental medical use of these treatments is linked to experimental social use: how women adopt them to respond to the pressures, anxieties and aspirations of work and intimate life. The paper argues that these experimental techniques are becoming morally authorized as routine management of women's health, integrated into mainstream Ob-Gyn healthcare, and subtly blurred with practices of cuidar-se (self-care) seen in Brazil as essential for modern femininity.


A genetic variant in the catechol-O-methyl transferase (COMT) gene is related to age-dependent differences in the therapeutic effect of calcium-channel blockers.
Hypertension is the leading risk factor for cardiovascular disease and one of the major health concerns worldwide. Genetic factors impact both the risk for hypertension and the therapeutic effect of antihypertensive drugs. Sex- and age-specific variances in the prevalence of hypertension are partly induced by estrogen. We investigated 6 single nucleotide polymorphisms in genes encoding enzymes involved in estrogen metabolism in relation to sex- and age-specific differences in the systolic and diastolic blood pressure (SBP and DBP) outcome under the treatment of diuretics, calcium-channel blockers (CCBs), angiotensin-converting-enzyme inhibitors, and angiotensin-receptor blockers (ARBs).We included 5064 subjects (age: 40-82) from the population-based CoLaus cohort. Participants were genotyped for the catechol-O-methyltransferase gene (COMT) variants rs4680, rs737865, and rs165599; the uridine-diphospho-glucuronosyltransferase 1A gene family (UGT1A) variants rs2070959 and rs887829; and the aromatase gene (CYP19A1) variant rs10046. Binomial and linear regression analyses were performed correcting for age, sex, body mass index, smoking, diabetes, and antihypertensive therapy to test whether the variants in focus are significantly associated with BP.All investigated COMT variants were strongly associated with the effect of diuretics, CCBs, and ARBs on SBP or DBP (P < .05), showing an additive effect when occurring in combination. After Bonferroni correction the polymorphism rs4680 (ValMet) in COMT was significantly associated with lower SBP in participants treated with CCBs (P = .009) with an especially strong impact in elderly individuals (age ≥ 70) alone (Δ = -14.08 mm Hg, P = .0005).These results underline the important role of estrogens and catecholamines in hypertension and the importance of genotype dependent, age-related adjustments of calcium-channel blocker treatment.


Cellular immunity to merozoite surface protein 2 (FC27 and 3D7) in Papua New Guinean children. Temporal variation and relation to clinical and parasitological status.
A prospective study in 207 children aged 0.5-15 years was carried out in a highly endemic area of Papua New Guinea to examine the relationship between cellular responses to Plasmodium falciparum merozoite surface protein 2 (MSP2) and malaria infection and morbidity. In vitro proliferation, IFN-gamma and IL-4 induction were measured against two recombinant proteins of MSP2, FC27 and 3D7 as well as against a form of the 3D7 MSP2 lacking the central repetitive sequences (d3D7). The prevalence of proliferative response was generally low, 6% for FC27, 9% for 3D7 and 11% for d3D7. A higher prevalence of IL-4 response was obtained being 27% for FC27, 34% for 3D7 and 30% for d3D7 while the prevalence of IFN-gamma response was 13%, 15% and 18%, respectively. There was no correlation between age and proliferative responses; in contrast cytokine production increased with age for all three antigens. When proliferation or stimulation of either cytokine was used to assess T-cell activation the frequency of responders increased to 39%, 47% and 46% for FC27, 3D7 and d3D7 respectively. Analysis of the relation of T cell responses to concurrent infection and morbidity showed that lymphoproliferative response only to d3D7 was significantly associated with parasitaemia; while lymphoproliferative responses to all 3 MSP2 antigens were highest in the group of clinical malaria cases. There was no significant correlation between proliferation or cytokine production to MSP2 and concurrent or subsequent malaria morbidity.


Impact of provider mailings on medication adherence by Medicare Part D members.
The Medicare 5-Star Rating System measures and provides incentive for improving Medicare Part D plans through a quality-based payment program. Adherence to medications for chronic conditions is key to the Star ratings. Our objective was to assess the impact of direct-to-provider letters on improving medication adherence. Members of a large US pharmacy benefits manager (PBM) who did not adhere to prescription of oral diabetes (antidiabetics), cholesterol-reducing (statins), or hypertension (renin angiotensin system [RAS] antagonists) drug therapy were identified from the prescriptions claims data of>600,000 continuously enrolled Medicare members. Nonadherence was defined by the Star ratings definition of proportion of days covered (PDC)<80%. The PBM sent letters to prescribing physicians of nonadherent members, requesting that they discuss adherence barriers and potential solutions with their patients. A historical control cohort was constructed from the PBM satisfying the same eligibility criteria as the intervention cohort. Both binary (≥80%) and continuous PDC measures were assessed as outcomes through multivariate logistic regression and difference-in-difference models, respectively. Final sample sizes were 21,044; 106,829; and 73,560 patients for antidiabetic, statin, and RAS antagonist use, respectively, with approximately equal number of intervention and control subjects in each drug class. Physician mailing was associated with 11%, 16%, and 7% higher odds of being adherent by members in antidiabetic, statin, and RAS antagonist cohorts, respectively (all P<.001). Within limitations of historical controls, physician mailing was associated with improved medication adherence. Physician mailing can be an impactful tool for improving medication adherence. II.


Possible adverse health effects of vitamin C and ascorbic acid.
Consensus from individual studies and several review articles is that consumption of supplemental vitamin C leads to no significant adverse health effects to humans in general. Individuals who have a history of kidney stone formation and those who experience iron overload should exercise caution before using supplemental vitamin C. Occasionally, individuals experience diarrhea or mild nausea. There is also the possibility that vitamin C taken simultaneously with other drugs may contribute to adverse health effects and that its interference in clinical laboratory tests will mask diagnosis of disease. Few controlled clinical trials exist that conclusively demonstrate the adverse health effects that humans may experience with supplemental vitamin C usage, and before definite conclusions can be made of the health hazards to humans, more clinical trials are required.


[Cigarette smoke extract down-regulates expression of histone deacetylase 2 and increases inflammatory cytokines releasing from murine C2C12 skeletal muscle myocytes].
To observe changes of inflammatory mediators, histone deacetylase 2 (HDAC2) and nuclear factor κB (NF-κB) in murine C2C12 skeletal muscle myocytes after exposed to cigarette smoke. Murine C2C12 skeletal muscle myocytes were cultured and treated with cigarette smoke extract (CSE). MTT assay was used to detect the effect of CSE on cell proliferation to determine appropriate concentration of CSE. The C2C12 cells cultured for 6-7 days were planted in six-well plates, and divided into control group, (6.25, 12.50, 25.0) mL/L CSE groups. The cells were cultured for 24 hours. The levels of interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α) in the supernatant were measured by ELISA. The mRNA level of HDAC2 was determined by real-time quantitative PCR. The protein level of HDAC2 was detected by Western blotting. HDAC2/NF-κB compound was determined by the method of co-immunoprecipitation. MTT assay showed that CSE at the concentration of 50 mL/L inhibited proliferation of C2C12 cells. After 24-hour treatment with CSE, IL-8 and TNF-α releasing from C2C12 cells increased and the level of HDAC2 mRNA and protein were reduced, which were CSE dose-dependent. Co-immunoprecipitation confirmed that HDAC2/NF-κB compound existed in the CSE-exposed C2C12 cells. CSE can down-regulate the expression of HDAC2 and increase inflammatory cytokines releasing from C2C12 cells.


Replication of It's Your Game…Keep It Real! in Southeast Texas.
Despite the recent efforts of the Office of Adolescent Health to replicate programs with demonstrated efficacy, there are still few evidence-based HIV, sexually transmitted infection (STI), and teen pregnancy prevention programs that have been replicated in "real-world" settings. To test the effectiveness of It's Your Game…Keep It Real! (IYG), an evidence-based STI and pregnancy prevention program for middle schools, the curriculum was implemented by teachers in urban and suburban middle schools in Southeast Texas from 2012 to 2015. IYG was evaluated using a group-randomized wait-list controlled effectiveness trial design in which 20 middle schools in nine urban and suburban school districts in Southeast Texas were randomized equally, using a multi-attribute randomization protocol, to either the intervention condition (received IYG) (n = 10 schools comprising 1936 eligible seventh graders) or the comparison condition (received usual care) (n = 10 schools comprising 1825 eligible seventh graders). All students were blinded to condition prior to administering the baseline survey. The analytic sample comprised 1543 students (n = 804, intervention; n = 739, comparison) who were followed from baseline (seventh grade) to the 24-month follow-up (ninth grade). Multilevel regression analyses were conducted to assess behavioral and psychosocial outcomes at follow-up. There were no significant differences in initiation of vaginal or oral sex between study conditions at follow-up. However, at 12-month follow-up, compared with students in the comparison condition, students in the intervention condition reported increased knowledge, self-efficacy, and perceived favorable norms related to HIV/STIs, condoms, and/or abstinence; decreased intentions to have sex; and increased intentions to use birth control. Knowledge outcomes were statistically significant at 24-month follow-up. This IYG effectiveness trial did not replicate the behavioral effects of the original IYG efficacy trials. However, it adds to the growing literature on the replication of evidence-based programs, and underscores the need to better understand how variations in implementation, setting, and measurement affect the behavioral impact of such programs.Clinical trial registration clinicaltrials.gov (NCT03533192).


Transient blockade of delta-like Notch ligands prevents allograft rejection mediated by cellular and humoral mechanisms in a mouse model of heart transplantation.
Rejection remains a major clinical challenge limiting allograft survival after solid organ transplantation. Both cellular and humoral immunity contribute to this complication, with increased recognition of Ab-mediated damage during acute and chronic rejection. Using a mouse model of MHC-mismatched heart transplantation, we report markedly protective effects of Notch inhibition, dampening both T cell and Ab-driven rejection. T cell-specific pan-Notch blockade prolonged heart allograft survival and decreased IFN-γ and IL-4 production by alloreactive T cells, especially when combined with depletion of recipient CD8(+) T cells. These effects were associated with decreased infiltration by conventional T cells and an increased proportion of regulatory T cells in the graft. Transient administration of neutralizing Abs specific for delta-like (Dll)1/4 Notch ligands in the peritransplant period led to prolonged acceptance of allogeneic hearts, with superior outcome over Notch inhibition only in T cells. Systemic Dll1/4 inhibition decreased T cell cytokines and graft infiltration, germinal center B cell and plasmablast numbers, as well as production of donor-specific alloantibodies and complement deposition in the transplanted hearts. Dll1 or Dll4 inhibition alone provided partial protection. Thus, pathogenic signals delivered by Dll1/4 Notch ligands early after transplantation promote organ rejection through several complementary mechanisms. Transient interruption of these signals represents an attractive new therapeutic strategy to enhance long-term allograft survival.


NKX3.1 activates cellular response to DNA damage.
The prostate-specific tumor suppressor homeodomain protein NKX3.1 is inactivated by a variety of mechanisms in the earliest phases of prostate carcinogenesis and in premalignant regions of the prostate gland. The mechanisms by which NKX3.1 exercises tumor suppression have not been well elucidated. Here, we show that NKX3.1 affects DNA damage response and cell survival after DNA damage. NKX3.1 expression in PC-3 prostate cancer cells enhances colony formation after DNA damage but has minimal effect on apoptosis. NKX3.1 also diminishes and regulates total cellular accumulation of gammaH2AX. Endogenous NKX3.1 in LNCaP cells localizes to sites of DNA damage where it affects the recruitment of phosphorylated ATM and the phosphorylation of H2AX. Knockdown of NKX3.1 in LNCaP cells attenuates the acute responses of both ATM and H2AX phosphorylation to DNA damage and their subnuclear localization to DNA damage sites. NKX3.1 expression enhances activation of ATM as assayed by autophosphorylation at serine 1981 and activation of ATR as assayed by phosphorylation of CHK1. An inherited mutation of NKX3.1 that predisposes to early prostate cancer and attenuates in vitro DNA binding was devoid of the ability to activate ATM and to colocalize with gammaH2AX at foci of DNA damage. These data show a novel mechanism by which a homeoprotein can affect DNA damage repair and act as a tumor suppressor.


Quality of Life in a Vitiligo Support Group.

BACKGROUND: No study has examined the impact of vitiligo support group membership on vitiligo patient quality of life (QoL).

OBJECTIVE: We sought to examine the QoL impact of vitiligo support groups by comparing QoL and associated patient characteristics between vitiligo patients who are and are not members of a vitiligo support group.

METHODS: Members of a Henry Ford Hospital-sponsored, Southeast Michigan Vitiligo Support Group were compared to non-member vitiligo patients recruited from a previous study cohort.17 Eligible patients were asked to complete the Dermatology Life Quality Index (DLQI) and a study-specific questionnaire designed to collect relevant patient characteristics.

RESULTS: The mean DLQI scores for the support group members and non-members were similar (7.1 ± 5.4 and 6.0 ± 6.5, respectively; P-value 0.2), despite the support group members reporting more severe overall disease and increased disease severity in exposed portions of the body. The African-American: Caucasian ratio and the prevalence of unemployment were both significantly higher among the support group participants. Small sample size may have limited the study's ability to demonstrate the differences between the support group participants and the controls.

CONCLUSIONS: The similar QoL despite an increased prevalence of poorer QoL indicators among the support group participants suggests a protective effect of support group membership.

J Drugs Dermatol. 2017;16(4):344-350.
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Evolution of Plant Architecture, Functional Diversification and Divergent Evolution in the Genus Atractocarpus (Rubiaceae) for New Caledonia.
The diversification of ecological roles and related adaptations in closely related species within a lineage is one of the most important processes linking plant evolution and ecology. Plant architecture offers a robust framework to study these processes as it can highlight how plant structure influences plant diversification and ecological strategies. We investigated a case of gradual evolution of branching architecture in Atractocarpus spp. (Rubiaceae), forming a monophyletic group in New Caledonia that has diversified rapidly, predominantly in rainforest understory habitats. We used a transdisciplinary approach to depict architectural variations and revealed multiple evolutionary transitions from a branched (Stone's architectural model) to a monocaulous habit (Corner's architectural model), which involved the functional reduction of branches into inflorescences. We propose an integrative functional index that assesses branching incidence on functional traits influencing both assimilation and exploration functions. We showed that architectural transitions correlate with ecologically important functional traits. Variation in ecologically important traits among closely relatives, as supported by the architectural analysis, is suggestive of intense competition that favored divergence among locally coexisting species. We propose that Pleistocene climatic fluctuations causing expansion and contraction of rainforest could also have offered ecological opportunities for colonizers in addition to the process of divergent evolution.