Expanding Family Medicine Scholarship to All Faculty: The Minnesota Model for Harmonizing Clinical Care, Education, and Research Missions.
The Department of Family Medicine and Community Health at the University of Minnesota engaged in a 5-year transformation to expand research and scholarship opportunities to all faculty. A harmonization framework was used to integrate the 3 missions of clinical care, education, and research to ensure that research and scholarship were an ongoing focus of the department. The key elements of our transformation included as follows: (1) a general culture of inquiry, (2) harmonized leadership, (3) training and mentoring, and (4) infrastructure and resources. Components of each of these elements were intentionally instituted simultaneously and iteratively across the 5 years to provide robust and sustainable research and scholarship opportunities for all faculty. Outputs and outcomes of the harmonized transformation indicated that clinical and research faculty publications increased, and the percentage of clinical faculty trained in research and scholarship skills increased across the 5 years. Important lessons learned during the harmonized transformation included the following: (1) key elements of the transformation need to be balanced as an ensemble, (2) cultural and organizational shifts take concerted effort and time, (3) embrace iteration: allow "bumps in the road" to propel the work forward, (4) transformation is financially feasible, (5) career research faculty can mutually benefit from clinical faculty engaging in scholarship, and (6) honor skepticism or disinterest and let people cultivate enthusiasm for research and scholarship rather than being forced.


Project ADAPT: a program to assess depression and provide proactive treatment in rural areas.
We describe and evaluate a project designed to pilot test an evidence-based clinical intervention for assessing and treating depression in older adults in rural primary care clinics. Project ADAPT-Assuring Depression Assessment and Proactive Treatment-utilized existing primary care resources to overcome barriers to sustainability experienced by similar projects. This multifaceted intervention, which was structured after the successful IMPACT (Improving Mood/Promoting Access to Collaborative Treatment) research intervention, used on-site geriatric depression specialists, clinic staff training, team collaboration, and depression practice guidelines to improve depression care for rural elders. IMPACT screening and assessment instruments and treatment protocols were modified for use by less highly trained staff already employed by the rural primary care clinics. Patient and provider depression educational materials and depression screening and monitoring protocols were provided by means of regional training sessions and phone contact. Evaluation data were collected by mail and phone surveys. Although Project ADAPT materials and training were initially developed for providers in rural primary care clinics, most participants came from long-term-care facilities, hospitals, home care, and public health and social service agencies. Forty-four sites sent 56 staff to Project ADAPT regional trainings, but many did not participate after the initial training. Participants who did continue reported that training improved geriatric depression screening and communication with the primary provider. Outcomes suggest that provider, patient, and system issues have to be addressed differently in rural areas to improve geriatric depression treatment in primary care settings.


The personal financial burden associated with idiopathic inflammatory myopathies.
The economic burden of idiopathic inflammatory myopathies (IIMs) within the US is underexplored. We hypothesized that IIMs patients experience considerable personal financial burden due to risks of multi-specialist visits, chronic long-term care, costs associated with disability, medical treatment, and overall high spending costs within the US healthcare system. We surveyed members of Myositis Support and Understanding (MSU) (response rate 4.7 %), and of the 470 survey participants that self-reported with diagnoses of IIMs, we assessed financial burden using two validated measures: (1) Financial Worry Score, and (2) Financial Burden Composite Score (FBCS). We determined factors associated with increased FBCS using logistic and Poisson regression respectively. High financial worry was endorsed by 202 participants (43 %) and the average FBCS ± SD was 1.8 ± 1.9. The odds of financial worry among participants with Medicaid is 3.016 times the odds of financial worry among participants without Medicaid (p = 0.011), and the odds of financial worry among participants with private high-deductible insurance is 3.216 times the odds of financial worry among participants who do not have private high-deductible insurance (p =< 0.001). Given the link between personal financial burden and potential effects on patient outcomes, it is essential for physicians to consider patient financial health when determining management or treatment courses. Identifying specific risk factors that can further exacerbate personal financial burden can help physicians identify vulnerable patients to reduce financial hardship.


Turning around financially distressed hospitals.
Just as a hospital's financial distress seldom results from a single mistake or catastrophe, its recovery involves a complex interplay between leadership, resources, and strategic action. To create a turnaround plan aimed at restoring a hospital to fiscal health, management must appoint a steering committee to oversee the effort; complete a thorough assessment of damage caused by financial problems; reduce operating expenses such as staffing and supply costs; and take steps to improve cash flow.


Chromosomal aberrations in workers exposed to low levels of benzene: association with genetic polymorphisms.
Benzene and its metabolites damage human lymphocytes, resulting in chromosomal aberrations and aneuploidy. Polymorphisms in the genes for benzene-metabolizing enzymes have been implicated in benzene-associated haematotoxicity. In this study, we examined the specificity of benzene-induced aneuploidy and the influence of genetic polymorphisms (GSTM1, GSTT1, GSTP1, NAT2, NQO1 and CYP2E1) on chromosomal aberrations. In total, 82 benzene-exposed workers from a coke oven plant and 76 matched controls were examined. The benzene concentration in the work-place air ranged from 0.014-0.743 p.p.m. (geometric mean 0.557 p.p.m.). Benzene exposure was associated with significant increases in both monosomy and trisomy of chromosomes 8 and 21. Translocations between chromosomes 8 and 21 [t(8:21)] were eight-fold more frequent in the high-level exposure group compared to the control group. Multiple regression analysis indicated that the frequencies of chromosome aberrations were significantly associated with benzene exposure and polymorphisms in the metabolic enzyme genes. A particular subset of genotypes, which included the GSTM1-null and GSTT1-null genotypes, the slow acetylator type of NAT2, a variant of the NQO1 genotype and the CYP2E1 DraI and RsaI genotypes, were either separately, or in combination, associated with increased frequencies of aneuploidy among the benzene-exposed individuals after adjustments for age, alcohol consumption and smoking. These results suggest that polymorphisms in the genes for benzene-metabolizing enzymes influence the susceptibility of individuals to chromosomal aberrations in relation to benzene exposure.


3-hydroxy-3-methylglutaryl coenzyme A reductase of Sulfolobus solfataricus: DNA sequence, phylogeny, expression in Escherichia coli of the hmgA gene, and purification and kinetic characterization of the gene product.
The gene (hmgA) for 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase (EC 184.108.40.206) from the thermophilic archaeon Sulfolobus solfataricus P2 was cloned and sequenced. S. solfataricus HMG-CoA reductase exhibited a high degree of sequence identity (47%) to the HMG-CoA reductase of the halophilic archaeon Haloferax volcanii. Phylogenetic analyses of HMG-CoA reductase protein sequences suggested that the two archaeal genes are distant homologs of eukaryotic genes. The only known bacterial HMG-CoA reductase, a strictly biodegradative enzyme from Pseudomonas mevalonii, is highly diverged from archaeal and eukaryotic HMG-CoA reductases. The S. solfataricus hmgA gene encodes a true biosynthetic HMG-CoA reductase. Expression of hmgA in Escherichia coli generated a protein that both converted HMG-CoA to mevalonate and cross-reacted with antibodies raised against rat liver HMG-CoA reductase. S. solfataricus HMG-CoA reductase was purified in 40% yield to a specific activity of 17.5 microU per mg at 50 degrees C by a sequence of steps that included heat treatment, ion-exchange chromatography, hydrophobic interaction chromatography, and affinity chromatography. The final product was homogeneous, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The substrate was (S)- not (R)-HMG-CoA; the reductant was NADPH not NADH. The Km values for HMG-CoA (17 microM) and NADPH (23 microM) were similar in magnitude to those of other biosynthetic HMG-CoA reductases. Unlike other HMG-CoA reductases, the enzyme was stable at 90 degrees C and was optimally active at pH 5.5 and 85 degrees C.


Expressing fear enhances sensory acquisition.
It has been proposed that facial expression production originates in sensory regulation. Here we demonstrate that facial expressions of fear are configured to enhance sensory acquisition. A statistical model of expression appearance revealed that fear and disgust expressions have opposite shape and surface reflectance features. We hypothesized that this reflects a fundamental antagonism serving to augment versus diminish sensory exposure. In keeping with this hypothesis, when subjects posed expressions of fear, they had a subjectively larger visual field, faster eye movements during target localization and an increase in nasal volume and air velocity during inspiration. The opposite pattern was found for disgust. Fear may therefore work to enhance perception, whereas disgust dampens it. These convergent results provide support for the Darwinian hypothesis that facial expressions are not arbitrary configurations for social communication, but rather, expressions may have originated in altering the sensory interface with the physical world.


Transcription of mouse mammary tumor virus: identification of a candidate mRNA for the long terminal repeat gene product.
We have examined an assortment of preneoplastic and neoplastic mouse mammary tissues for the presence of an mRNA which could encode the putative long terminal repeat gene product of mouse mammary tumor virus. We report here the detection of a novel mouse mammary tumor virus-specific, polyadenylic acid-containing transcript in certain preneoplastic and neoplastic mammary tissue of BALB/c mice. The molecule is 1.6 kilobases in length and contains sequences from the transcriptional leader and the U3 region of the proviral DNA. The upstream terminus of the 3' information lies 75 to 80 nucleotides from the beginning of the long terminal repeat open reading frame, in close proximity to a consensus splice acceptor in the DNA. The transcript was detected in hormonally or chemically induced neoplastic, preneoplastic, and lactating mammary tissue of BALB/c mice, but not in preneoplastic or tumor tissue induced by exogenous viruses in any strain of mice examined. This implies that the RNA we observed is transcribed from an endogenous provirus template.


Identification of proteoglycans as the APRIL-specific binding partners.
B cell activating factor of the tumor necrosis factor (TNF) family (BAFF) and a proliferation-inducing ligand (APRIL) are closely related ligands within the TNF superfamily that play important roles in B lymphocyte biology. Both ligands share two receptors--transmembrane activator and calcium signal--modulating cyclophilin ligand interactor (TACI) and B cell maturation antigen (BCMA)--that are predominantly expressed on B cells. In addition, BAFF specifically binds BAFF receptor, whereas the nature of a postulated APRIL-specific receptor remains elusive. We show that the TNF homology domain of APRIL binds BCMA and TACI, whereas a basic amino acid sequence (QKQKKQ) close to the NH2 terminus of the mature protein is required for binding to the APRIL-specific "receptor." This interactor was identified as negatively charged sulfated glycosaminoglycan side chains of proteoglycans. Although T cell lines bound little APRIL, the ectopic expression of glycosaminoglycan-rich syndecans or glypicans conferred on these cells a high binding capacity that was completely dependent on APRIL's basic sequence. Moreover, syndecan-1-positive plasma cells and proteoglycan-rich nonhematopoietic cells displayed high specific, heparin-sensitive binding to APRIL. Inhibition of BAFF and APRIL, but not BAFF alone, prevented the survival and/or the migration of newly formed plasma cells to the bone marrow. In addition, costimulation of B cell proliferation by APRIL was only effective upon APRIL oligomerization. Therefore, we propose a model whereby APRIL binding to the extracellular matrix or to proteoglycan-positive cells induces APRIL oligomerization, which is the prerequisite for the triggering of TACI- and/or BCMA-mediated activation, migration, or survival signals.


In vivo safety and biodistribution profile of Klotho-enhanced human urine-derived stem cells for clinical application.
Urine-derived stem cells (UDSCs) can be easily isolated from urine and possess excellent stem cell characteristics, making them a promising source for cell therapeutics. Due to their kidney origin specificity, UDSCs are considered a superior therapeutic alternative for kidney diseases compared to other stem cells. To enhance the therapeutic potential of UDSCs, we developed a culture method that effectively boosts the expression of Klotho, a kidney-protective therapeutic factor. We also optimized the Good Manufacturing Practice (GMP) system to ensure stable and large-scale production of clinical-grade UDSCs from patient urine. In this study, we evaluated the in vivo safety and distribution of Klotho-enhanced UDSCs after intravenous administration in accordance with Good Laboratory Practice (GLP) regulations. Mortality and general symptoms were continuously monitored throughout the entire examination period. We evaluated the potential toxicity of UDSCs according to the administration dosage and frequency using clinical pathological and histopathological analyses. We quantitatively assessed the in vivo distribution and retention period of UDSCs in major organs after single and repeated administration using human Alu-based qPCR analysis. We also conducted long-term monitoring for 26 weeks to assess the potential tumorigenicity. Klotho-enhanced UDSCs exhibited excellent homing potential, and recovered Klotho expression in injured renal tissue. Toxicologically harmful effects were not observed in all mice after a single administration of UDSCs. It was also verified that repeated administration of UDSCs did not induce significant toxicological or immunological adverse effects in all mice. Single and repeated administrated UDSCs persisted in the blood and major organs for approximately 3 days and cleared in most organs, except the lungs, within 2 weeks. UDSCs that remained in the lungs were cleared out in approximately 4-5 weeks. There were no significant differences according to the variation of sex and administration frequency. The tumors were found in the intravenous administration group but they were confirmed to be non-human origin. Based on these results, it was clarified that UDSCs have no tumorigenic potential. Our results demonstrate that Klotho-enhanced UDSCs can be manufactured as cell therapeutics through an optimized GMP procedure, and they can be safely administered without causing toxicity and tumorigenicity.


Severe aortic regurgitation resulting from a downward displacement of anterior aortic annulus and fibrous strands in the bicuspid aortic valve.
Aortic regurgitation is occasionally caused by fibrous strands and more rarely by downward displacement of the aortic annulus. The present report describes an 18-year-old man with aortic regurgitation resulting from an anterior-posterior type of bicuspid aortic valve with fibrous strands and downward displacement of the anterior aortic annulus. A pair of fibrous strands at the anterior cusp of the bicuspid valve lifted the free margin of the cusp, and the anterior cusp originated from the intraventricular septum. We considered that the aortic regurgitation was due to poor coaptation of the cusps because of these two conditions. After resection of the cusps and the strands, the aortic valve was replaced at the intra-annular position.


A joint approach to treating dental phobia: a re-evaluation of a collaboration between community dental services and specialist psychotherapy services ten years on.
To audit the records of a group of patients who had previously benefited from cognitive behavioural therapy (CBT) for dental phobia. To ascertain if they had returned to the use of intravenous (IV) sedation to facilitate dental treatment. Ten years ago these patients were routinely requiring IV sedation to facilitate dental treatment due to severe dental phobia. Sixty patients entered the original pilot project. Of those, 30 were offered CBT and 21 attended. Twenty of those patients (95.2%) were subsequently able to have dental treatment without IV sedation. In this follow-up study the electronic records of 19 of the 20 patients who had originally been successful with CBT were re-audited. Our purpose was to see if there was any record of subsequent IV sedation administration in the intervening ten years. Of the 19 successful CBT patients available to follow-up, 100% had not received IV sedation since the study ten years ago. This may suggest the initial benefit of CBT has endured over the ten-year period. This study indicates that the use of CBT for patients with dental phobia proves beneficial not only in the initial treatment but that the benefits may endure over time. This results in a significant reduction in health risks to the patient from repeated IV sedation. It may also translate into significant financial savings for dental care providers. Our evidence for CBT as treatment for dental phobia suggests dental services should be implementing this approach now rather than pursuing further research.


Weakly activated core neuroinflammation pathways were identified as a central signaling mechanism contributing to the chronic neurodegeneration in Alzheimer's disease.
Neuroinflammation signaling has been identified as an important hallmark of Alzheimer's disease (AD) in addition to amyloid β plaques (Aβ) and neurofibrillary tangles (NFTs). However, the molecular mechanisms and biological processes of neuroinflammation remain unclear and have not well delineated using transcriptomics data available. Our objectives are to uncover the core neuroinflammation signaling pathways in AD using integrative network analysis on the transcriptomics data. From a novel perspective, i.e., investigating weakly activated molecular signals (rather than the strongly activated molecular signals), we developed integrative and systems biology network analysis to uncover potential core neuroinflammation signaling targets and pathways in AD using the two large-scale transcriptomics datasets, i.e., Mayo Clinic (77 controls and 81 AD samples) and ROSMAP (97 controls and 260 AD samples). Our analysis identified interesting core neuroinflammation signaling pathways, which are not systematically reported in the previous studies of AD. Specifically, we identified 7 categories of signaling pathways implicated on AD and related to virus infection: immune response, x-core signaling, apoptosis, lipid dysfunctional, biosynthesis and metabolism, and mineral absorption signaling pathways. More interestingly, most of the genes in the virus infection, immune response, and x-core signaling pathways are associated with inflammation molecular functions. The x-core signaling pathways were defined as a group of 9 signaling proteins: MAPK, Rap1, NF-kappa B, HIF-1, PI3K-Akt, Wnt, TGF-beta, Hippo, and TNF, which indicated the core neuroinflammation signaling pathways responding to the low-level and weakly activated inflammation and hypoxia and leading to the chronic neurodegeneration. It is interesting to investigate the detailed signaling cascades of these weakly activated neuroinflammation signaling pathways causing neurodegeneration in a chronic process, and consequently uncover novel therapeutic targets for effective AD treatment and prevention. The potential core neuroinflammation and associated signaling targets and pathways were identified using integrative network analysis on two large-scale transcriptomics datasets of AD.


Epithelially Restricted Interferon Epsilon Protects Against Colitis.
Type I interferon (T1IFN) signalling is crucial for maintaining intestinal homeostasis. We previously found that the novel T1IFN, IFNε, is highly expressed by epithelial cells of the female reproductive tract, where it protects against pathogens. Its function has not been studied in the intestine. We hypothesize that IFNε is important in maintaining intestinal homeostasis. We characterized IFNε expression in mouse and human intestine by immunostaining and studied its function in the dextran sulfate sodium (DSS) colitis model using both genetic knockouts and neutralizing antibody. We demonstrate that IFNε is expressed in human and mouse intestinal epithelium, and expression is lost in inflammation. Furthermore, we show that IFNε limits intestinal inflammation in mouse models. Regulatory T cell (Treg) frequencies were paradoxically decreased in DSS-treated IFNε-/- mice, suggesting a role for IFNε in maintaining the intestinal Treg compartment. Colitis was ameliorated by transfer of wild-type Tregs into IFNε-/- mice. This demonstrates that IFNε supports intestinal Treg function. Overall, we have shown IFNε expression in intestinal epithelium and its critical role in gut homeostasis. Given its known role in the female reproductive tract, we now show IFNε has a protective role across multiple mucosal surfaces.


Marrow stimulation procedures for high-grade cartilage lesions during surgical repair of medial meniscus root tear yielded suboptimal outcomes, whilst small lesions showed surgical eligibility.
To investigate the surgical outcomes of arthroscopic pull-out repair for medial meniscus root tear (MMRT) combined with the marrow stimulation procedures (MSP) for accompanying high-grade cartilage lesions. Patients who underwent arthroscopic pull-out repair for MMRT between 2010 and 2019 were retrospectively reviewed. Patients who had at least 3 years of follow-up were included and classified into two groups according to whether MSP (microfracture or microdrilling) were performed on cartilage lesions in the medial tibiofemoral joint (group 1, patients with International Cartilage Repair Society [ICRS] grade 0-3a lesions and did not undergo MSP; group 2, patients with ICRS grade 3b-3d lesions and underwent MSP). Comparative analyses, including non-inferiority trials, were conducted between groups for subjective and objective outcomes. In addition, group 2 was further divided into two subgroups according to cartilage lesion size and compared with group 1 (group S, ≤ 2.0 cm2; group L, > 2.0 cm2). A total of 94 patients were included (group 1, 68 patients; group 2, 26 patients). There were no significant differences in clinical scores at postoperative 3 years and final follow-up between groups 1 and 2, but group 2 failed to satisfy the non-inferiority criteria compared to group 1 overall. In objective outcomes, group 2 did not meet the non-inferiority criteria for the rate of osteoarthritis progression compared to group 1, and it also showed a significantly higher proportion of high-grade osteoarthritis at final follow-up (P = 0.044) and a higher degree of osteoarthritis progression than group 1 (P = 0.03 for pre- to postoperative 3 years, and P = 0.006 for pre- to final follow-up). In additional evaluations comparing the subgroups of group 2 and group 1, group S showed relatively favourable results compared to group L in objective outcomes at final follow-up. Patients who underwent arthroscopic pull-out repair for MMRT combined with MSP for accompanying high-grade cartilage lesions showed suboptimal outcomes compared to those with no or low-grade lesions at mid-term follow-up. High-grade cartilage lesions ≤ 2.0 cm2 may be candidates for the surgical repair of MMRT if MSP are performed, but those with larger lesions may require alternative treatment strategies. III.


Optimal conditions for rearing the tadpole shrimp, Triops longicaudatus (Notostraca:Triopsidae), a biological control agent against mosquitoes.
Tadpole shrimp (TPS) were reared at 15, 20, 25, and 30 degrees C in the laboratory. Size of TPS was temperature and age dependent with more rapid development at warmer temperatures. Survivorship was inversely related to rearing temperature. Mean age at death was 24.2, 19.2, 14.3, and 11.8 days at 15, 20, 25, and 30 degrees C, respectively. Rearing temperature (excluding 15 degrees C) did not affect lifetime fecundity, but larger TPS laid more eggs than smaller ones. Tadpole shrimp began ovipositing earlier at higher temperatures, and at a smaller size than their counterparts in lower temperatures. Mean age at reproductive maturity was 18.8, 13.1, and 10.2 days and mean carapace length was 10.8, 11.0, and 10.3 at 20, 25, and 30 degrees C, respectively. Embryogenesis required a minimum of 3 days for completion. Hatching rates during the first hydration decreased with increasing egg batch number produced by individuals, ranging from a mean of 74% for the first batch to 31% for the 5th batch. Cumulative hatching rates of eggs after 2 hydrations were consistent across temperatures and egg batches (79 +/- 2%).


Substantial cardiac parasympathetic activity exists during heavy dynamic exercise in dogs.
We investigated the extent of functional parasympathetic and sympathetic activity to the heart at rest and during mild to heavy dynamic exercise in conscious dogs. The animals were chronically instrumented to monitor mean arterial pressure (MAP), heart rate (HR), and terminal aortic blood flow (TAQ) and trained to run on a motor-driven treadmill. MAP, HR, and TAQ were monitored at rest and during steady-state dynamic exercise ranging from mild [3.2 kilometers per hour (kph), 0% grade] to heavy exercise (8 kph, 15% grade). Experiments were performed before and after blocking the effects of either the parasympathetic nerves (atropine 0.2 mg/kg i.v.) or sympathetic nerves (atenolol 2.0 mg/kg i.v.) to the heart. In addition, blood samples were taken at rest and at steady state during exercise, and plasma levels of vasopressin and renin activity were assessed. At rest and during all levels of exercise, muscarinic cholinergic receptor blockade caused a marked increase in HR over control (saline treated) levels with little effect on MAP or TAQ. beta-Adrenergic receptor blockade had no significant effect on HR at rest and during mild exercise. At moderate to heavy workloads, beta-receptor blockade significantly reduced MAP, HR, and TAQ and increased plasma vasopressin levels. We conclude that, even during heavy dynamic exercise, significant functional parasympathetic tone to the heart exists. Thus, over a wide range of exercise workloads, HR is under the tonic control of both sympathetic and parasympathetic nerves.


Appropriate patient selection in the management of common bile duct stones: when not to do ERCP.
Background. Magnetic resonance cholangiopancreatography (MRCP) is noninvasive and accurate for diagnosing intra common bile duct stones (ICSs). However, given limited access, routine utilisation for investigating all patients with gallstone disease is neither practical nor cost-effective. Conversely, many individuals proceed directly to endoscopic retrograde cholangiopancreatography (ERCP), an invasive test with appreciable complications. Aim. Identify factors associated with ICS in order to improve risk-stratification for MRCP/ERCP. Methods. All patients having undergone cholecystectomy between November 2007 and October 2008 were reviewed. High-risk features for ICS were predefined, and their true presence confirmed by ERCP or intraoperative cholangiogram. Multivariate logistic regression was performed on candidate risk features. Results. Of 231 patients, 10.4% had ICS. Defining a high-risk group with "both" biochemical and ultrasound risk factors predicted ICS with 92% specificity and also bore strong association (OR 8.88). However, isolated hyperbilirubinaemia, ultrasound impression of CBD stones, and clinical risk factors did not (OR 1.10, 0.97, and 1.26). Normal liver biochemistry and normal ultrasound had a NPV of 99.5% for ICS. Conclusions. Ultrasound impression of CBD calculi without ductal dilatation is not predictive of ICS. Patients with normal liver biochemistry and normal CBD diameter on ultrasound are unlikely to have ICS and should not proceed to ERCP.


PLURIPOTENT STEM CELL FUNCTION OF THE MOUSE MARROW "LYMPHOCYTE".
Bone marrow from normal and polycythemic mice was filtered through glasswool columns to remove cells other than "lymphocytes." For a given number of nucleated cells, filtered marrow was more efficient than the original marrow in repopulating the spleen of an isogenic recipient previously exposed to lethal irradiation. The proliferative capacity of both the filtered and unfiltered marrow suspensions appeared to be a constant function of the number of small and medium "lymphocytes" present and not of any other cell type.


Small-cell lung cancer, paraneoplastic cerebellar degeneration and the Lambert-Eaton myasthenic syndrome.
Several cancers, especially lung, ovarian and breast, can cause paraneoplastic cerebellar degeneration. The presence of different antineuronal antibodies associated with different cancers and paraneoplastic cerebellar degeneration suggests that several immunological mechanisms may result in the same neurological disorder. In patients with small-cell lung cancer, paraneoplastic cerebellar degeneration may occur with or without Hu antineuronal antibodies (HuAb), indicating that patients with the same tumour can develop paraneoplastic cerebellar degeneration by different immunological mechanisms. Furthermore, paraneoplastic cerebellar degeneration sometimes occurs in association with the Lambert-Eaton myasthenic syndrome. In order to try to understand the clinical implication of antineuronal antibodies in patients with small-cell lung cancer, we examined the serum of 57 patients with presenting symptoms of paraneoplastic cerebellar degeneration for the presence of HuAb and P/Q- and N-type voltage-gated calcium channel antibodies. Patients with paraneoplastic cerebellar degeneration who were HuAb positive were compared with HuAb negative patients with respect to neurological symptoms, course of the neurological disorder, response to treatment, tumour prognosis, pathological findings, and cause of death. The tumour outcome and serological findings of these patients were also compared with those of 109 small-cell lung cancer patients without paraneoplastic syndromes of the CNS. Titres of HuAb were classified as 'high' (immunoblot titre > 1:10,000) or 'low' (< 1:10,000), the latter similar to the antibody titres detected in some small-cell lung cancer patients without paraneoplastic symptoms. Twenty-five patients with paraneoplastic cerebellar degeneration (44%) had high titres of HuAb, four (7%) had low titres of HuAb, and 28 (49%) were HuAb negative; for clinical comparisons with the patients with high titres of HuAb, the four patients with low antibody titres were included in the HuAb negative cohort. None of the 109 small-cell lung cancer patients without paraneoplastic symptoms had high titres of HuAb. The presence of high titres of HuAb defined a subset of patients who differed from the HuAb negative paraneoplastic cerebellar degeneration cohort, HuAb positive patients were more likely to be female (P < 0.01), to have multifocal neurological disease (brainstem encephalopathy and sensory neuropathy being common extracerebellar manifestations) (P < 0.002), and be severely disabled (P < 0.005). A total of nine patients (16%) from both paraneoplastic cerebellar degeneration groups developed electrophysiologically confirmed Lambert-Eaton myasthenic syndrome. Seven of these nine patients had serum available for P/Q-type voltage-gated calcium channel antibody testing and all seven were positive. In addition, 20% of HuAb negative paraneoplastic cerebellar degeneration patients without clinically identified Lambert-Eaton myasthenic syndrome had P/Q-type voltage-gated calcium channel antibodies, while only 2% of small-cell lung cancer patients without paraneoplastic symptoms had these antibodies. Treatment of the tumour and/or immunomodulation did not alter the course of paraneoplastic cerebellar degeneration, but improved Lambert-Eaton myasthenic syndrome symptoms. At the time of death, in 60% of HuAb positive and 20% of HuAb negative paraneoplastic cerebellar degeneration patients, the tumour was either not evident or localized to the chest (P < 0.007); neurological disease was the cause of death of 65% HuAb positive paraneoplastic cerebellar degeneration and 10% HuAb negative paraneoplastic cerebellar degeneration patients (P < 0.001). (ABSTRACT TRUNCATED)


Determination of Wolbachia genome size by pulsed-field gel electrophoresis.
Genome sizes of six different Wolbachia strains from insect and nematode hosts have been determined by pulsed-field gel electrophoresis of purified DNA both before and after digestion with rare-cutting restriction endonucleases. Enzymes SmaI, ApaI, AscI, and FseI cleaved the studied Wolbachia strains at a small number of sites and were used for the determination of the genome sizes of wMelPop, wMel, and wMelCS (each 1.36 Mb), wRi (1.66 Mb), wBma (1.1 Mb), and wDim (0.95 Mb). The Wolbachia genomes studied were all much smaller than the genomes of free-living bacteria such as Escherichia coli (4.7 Mb), as is typical for obligate intracellular bacteria. There was considerable genome size variability among Wolbachia strains, especially between the more parasitic A group Wolbachia infections of insects and the mutualistic C and D group infections of nematodes. The studies described here found no evidence for extrachromosomal plasmid DNA in any of the strains examined. They also indicated that the Wolbachia genome is circular.


Genetic alterations in gastric adenomas of intestinal and foveolar phenotypes.
Gastric adenomas are unusual neoplasms that can constitute one of the direct precursors to gastric adenocarcinoma. Most gastric adenomas are comprised of polypoid projections of dysplastic epithelium with at least focal intestinal-type differentiation (containing goblet cells and/or Paneth cells), whereas adenomas comprised entirely of dysplastic foveolar-type epithelium are rare. It has been shown that nearly all intestinal-type adenomas arise in association with background intestinal metaplasia and gastric atrophy, approximately 40% harbor high-grade dysplasia, and nearly one fourth progress to adenocarcinoma. In contrast, foveolar-type adenomas tend to occur in otherwise normal, nonatrophic gastric mucosa and rarely harbor high-grade dysplasia or carcinoma. Potential differences in the genetic alterations between intestinal-type and foveolar-type gastric adenomas have not been systematically studied. We investigated 11 intestinal-type and 7 foveolar-type gastric adenomas (all from patients without familial adenomatous polyposis) for alterations in APC (using 5q allelic loss assays and direct DNA sequencing of the mutation cluster region), beta-catenin (using direct DNA sequencing of the phosphorylation region in exon 3), K-ras (using direct DNA sequencing of codons 12 and 13), and microsatellite instability (MSI; using fluorescent-based PCR amplification of a standard panel of 5 microsatellite markers). Overall, 10 of 11 (91%) intestinal-type adenomas harbored at least one detectable genetic alteration, whereas only 3 of 7 (43%) of foveolar-type adenomas did (P =.047). However, no statistically significant differences in any particular genetic alteration were found. Among intestinal-type adenomas, APC alterations were present in seven (64%), high-level MSI in three (27%), and K-ras mutations in two (18%). Among foveolar-type adenomas, APC alterations were present in three (43%) and a K-ras mutation in one of six amplifiable polyps (17%). Neither APC nor MSI correlated with the size of the adenoma, but K-ras mutations were found only in lesions of > or = 1 cm. beta-catenin mutations were not present in any gastric adenoma, irrespective of the presence or absence of APC alterations. These results suggest that the types and frequencies of genetic alterations occurring in gastric and colorectal adenomas are similar. Although intestinal-type and foveolar-type gastric adenomas display divergent biologic behavior, the specific genetic events accounting for these differences in morphology and biologic behavior are unclear.


Inflammatory markers are associated with quality of life, physical activity, and gait speed but not sarcopenia in aged men (40-79 years).
Age-related chronic low-grade inflammation (inflammaging) is one of the proposed mechanisms behind sarcopenia. However, findings regarding inflammatory markers in sarcopenic older adults are conflicting. This study aimed to determine the association between inflammatory markers, prevalent as well as incident sarcopenia, sarcopenia-defining parameters, quality of life (QoL), and physical activity in middle-aged and older men. Men aged 40-79 years (mean 59.66 ± 11.00y) were recruited from population registers in eight European centres for participation in the European Male Aging study (EMAS). Subjects were assessed at baseline (2003-2005) and again after a median follow-up of 4.29 years. In 2577 participants, associations between baseline inflammatory markers [high-sensitive C-reactive protein (hs-CRP), white blood cell count (WBC), albumin] and baseline physical activity (PASE) and QoL (SF-36) were analysed. In the Leuven and Manchester cohort (n = 447), data were available on muscle mass (whole-body dual X-ray absorptiometry) and strength. In this subgroup, cross-sectional associations between baseline inflammatory markers and sarcopenia-defining parameters (handgrip strength, chair stand test, appendicular lean mass, and gait speed) and prevalent sarcopenia were examined. In a further subgroup (n = 277), associations with knee extensor strength were explored. Longitudinally, predictive value of baseline inflammation on functional decline, physical activity, QoL, and incident sarcopenia was examined. Subgroup analyses were performed in subgroups with chronic inflammation and stratified by age. Linear and logistic regressions were used, adjusted for age, body mass index, centre, and smoking. At baseline, hs-CRP and WBC were negatively associated with PASE score (hs-CRP: β = -7.920, P < 0.001; and WBC: β = -4.552, P < 0.001) and the physical component score of SF-36 (hs-CRP: β = -1.025, P < 0.001; and WBC: β = -0.364, P < 0.001). Baseline WBC levels were negatively associated with gait speed (β = -0.013; P = 0.025), quadriceps isometric 90° (β = -5.983; P = 0.035) and isokinetic 60°/s peak torque/body weight (β = -5.532; P = 0.027). The prevalence of sarcopenia at baseline was 18.1% (n = 81). Of those without sarcopenia at baseline, 64 (18.6%) satisfied criteria for sarcopenia at follow-up. There were no significant associations between baseline inflammatory markers and either prevalent or incident sarcopenia, or change in level of sarcopenia-defining parameters between baseline and follow-up. In middle-aged and older men, hs-CRP and WBC were negatively associated with QoL and PASE scores, while WBC was negatively associated with gait speed and knee strength. Associations with hs-CRP remained significant in all ages, whereas WBC levels were only associated with PASE, gait speed and knee strength in older adults (60-79 years). Baseline inflammatory markers (hs-CRP, WBC and albumin) did not predict functional decline, decline in physical activity, decreased QoL or incident sarcopenia.


Optimized conditions for simultaneous determination of opiates, cocaine and benzoylecgonine in hair samples by GC--MS.
The present paper describes a qualitative and quantitative method for the simultaneous detection of opiates, cocaine and benzoylecgonine from human hair samples. Every step of the analytical procedure was studied to find the optimized conditions. Nine different incubation systems were examined. The influence of different pH values of samples on the isolation of analytes from the incubation media by Bond Elut cartridges and the stability of the compounds of interest in the different incubation media and conditions were investigated. The extracting power of different incubation media was studied as well. The phosphate buffer 0.1 N at pH 5 was chosen as the extraction medium in an optimized procedure for simultaneous determination of opiates, cocaine and benzoylecgonine in hair samples. The method developed was validated. Recoveries were 90% for morphine (M), 81% for 6-monoacetylmorphine (6-AM), 90% for codeine (CD), 86% for cocaine (C) and 90% for benzoylecgonine (BE). Relative standard deviation for inter-day precision was better than 12%. The limits of detection resulted as 0.05 ng/mg for M and C, as 0.08 for 6-AM and as 0.2 ng/mg for BE. Forty hair samples collected from drug abusers admitted to centers for detoxification treatment were analyzed obtaining 23 positive results for opiates and/or cocaine. Twelve hair specimens longer than 10 cm were analyzed following a sectional approach. In the six positive cases, it was interesting to find that the 6-AM/M ratio generally decreased for each sample from the proximal segment to the distal segments. Moreover, the 6-AM/M ratio was generally lower than 1 in the intermediate and distal segments.


[Hemodynamic mechanisms of the hypotensive effect of cobalt in anesthetized rats].
Hemodynamic mechanisms of Co(2+)-evoked hypotension were studied in pentobarbital--anesthetised male Wistar rats. Bolus i.v. administration of cobalt sulphate (30, 90 and 270 mg/kg of Co) evoked dose-dependent hypotension followed by tachycardia and increase in left ventricular contractility index (dP/dt/P). Administration of beta-adrenoblocker propranolol (2 mg/kg) attenuated Co evoked (270 ug/kg) tachycardia and unmasked its cardiodepressive effects (dP/dt/P decreased by 11% and end-diastolic pressure increased by 55%). Cardiodepressive Co2+ effects were prevented by administration of verapamil (0.4 mg/kg). In the second group hemodynamic mechanisms of Co-evoked hypotension were studied with radioactive microspheres. Co2+ infusion (55 mg/kg) lowered blood pressure (in average by -11%) by decreasing total peripheral resistance (-25%). Cardiotoxic effects were obtained with a ++larger dose 80 mg/kg/min. It is concluded that Co decreased blood pressure by vasodilatator action and its cardiotoxic effects attenuated by sympathetic counterregulation and may be prevented by verapamil.


Anti-inflammatory effect of different PRGF formulations on cutaneous surface.
Cutaneous autoimmune and inflammatory diseases are a major burden of global disease and many lack effective treatments that can derive in different dermatoses like atopic dermatitis. Despite the increase prevalence and the high health-care costs worldwide, the heterogeniety and multifactoriality of these diseases mean that effective treatment options are scarce. Plasma rich in growth factors (PRGF) technology could be an alternative approach that may help in the management of this cutaneous condition. The aim of this study was to assess the effect of two different PRGF formulations (just activated and autologous topical serum (ATS)) for the management of skin inflammation. Additionally, ATS was assessed over two patients suffering from radiotherapy induced dermatitis. Human organotypic skin explant cultures (hOSECs) were used as human skin models. To induce atopic dermatitis-like conditions, skin explants were treated with both interleukin-4 (IL-4) and interleukin-13 (IL-13). PRGF and ATS were intradermally and topically applied, respectively. Metabolic activity, reactive oxigen species (ROS), necrosis and inflammatory cytokine production were determined. Both PRGF formulations increased tissue viability and significantly reduced the excessive free radical accumulation and the cutaneous cytokine production such as TNF-α and IL-1β. Case reports showed a positive response after ATS treatment in terms of skin quality improvement, local erythema decrease and burning and itching amelioration. The oedema, swelling and desquamation caused by radiation induced dermatitis was also reduced and the patients referred ceased pruritus and pain. This preliminary study suggests that PRGF might aid in the management of inflammatory skin conditions.


Physical Activity Levels of Chilean Children in a National School Intervention Programme. A Quasi-Experimental Study.
Background. Recess is a great opportunity to interrupt sedentary behaviour and increase moderate-to-vigorous physical activity (MVPA) in schoolchildren. This quasi-experimental study aimed to compare the levels of physical activity (PA) during the school day of children in a school intervention programme vs. those in a control group, and to determine compliance with MVPA recommendations. Methods. A sample of 154 schoolchildren (6-12 years old) was obtained from several schools (70 with the intervention and 84 controls). This programme was structured with a duration of 90 min/session and performed three times/week. PA levels were recorded with triaxial accelerometers during the school day: during recess, during a PA session or physical education session (PE), and during lunchtime. No pre-intervention evaluation was performed. Results. The MVPA of the control group was higher than that of the intervention group during the first recess (p < 0.001). None of the groups complied with the recommendations for steps during the PA or PE sessions. During the PA session, sedentary time was lower and MVPA was higher, in the intervention group than in the control group. Fifty percent of the children from the intervention group complied with the MVPA recommendations, vs. 22.7% of those in the control group. Conclusions. The schoolchildren in the intervention group performed more MVPA than those in the control group. Future interventions could include other periods, such as recess and lunchtime, which are opportunities for improving the MVPA levels of schoolchildren.


Melatonin and brain inflammaging.
Melatonin is known to possess several properties of value for healthy aging, as a direct and indirect antioxidant, protectant and modulator of mitochondrial function, antiexcitotoxic agent, enhancer of circadian amplitudes, immune modulator and neuroprotectant. It is levels tend to decrease in the course of senescence and are more strongly reduced in several neurodegenerative disorders, especially Alzheimer's disease, and in diseases related to insulin resistance such as diabetes type 2. Although the role of melatonin in aging and age-related diseases has been repeatedly discussed, the newly emerged concept of inflammaging, that is, the contribution of low-grade inflammation to senescence progression has not yet been the focus of melatonin research. This review addresses the multiple protective actions of melatonin and its kynuramine metabolites that are relevant to the attenuation of inflammatory responses and progression of inflammaging in the brain, i.e. avoidance of excitotoxicity, reduction of free radical formation by support of mitochondrial electron flux, prevention of NADPH oxidase activation and suppression of inducible nitric oxide synthase, as well as downregulation of proinflammatory cytokines. The experimental evidence is primarily discussed on the basis of aging and senescence-accelerated animals, actions in the immune system, and the relationship between melatonin and sirtuins, having properties of aging suppressors. Sirtuins act either as accessory components or downstream factors of circadian oscillators, which are also under control by melatonin. Inflammaging is assumed to strongly contribute to neurodegeneration of the circadian master clock observed in advanced senescence and, even more, in Alzheimer's disease, a change that affects countless physiological functions.


Nursing Student Coaches for Emergency Department Super Utilizers.
The Super Utilizer Project is a collaboration between an integrated health care organization and a baccalaureate school of nursing. The goal is to help the super utilizers access appropriate health care resources through the use of a student coach model. The student coaches provide clients with disease prevention and health promotion education and assistance specific to the clients' unique health care needs. The tenets of the theoretical frameworks of servant leadership, caring theory, and transtheoretical model of behavior change are applied throughout this project. First-year outcomes for the project were encouraging, with the total number of visits for 23 super utilizers to the emergency department decreasing from 710 annually to a projected 510 annually, and second-semester visits for 20 clients decreased from 634 to a projected 462. The students experience complex client care and collaboration with an interdisciplinary team reducing nonemergent emergency department super utilizer visits. [J Nurs Educ. 2017;56(1):27-30.].


Place visualization: conventional or unconventional practice?
This paper explores the traditional notion that two discrete forms of medicine exist. One being bio-medically led, the other being holistic in origin. A qualitative study of place visualization in nursing is used to illustrate how health care may not always fit neatly into one of these paradigms. It is suggested that, through the use of this practice, an organically occurring therapeutic intervention emerges that includes core elements of each category. Based on the findings of this small scale inductive study, it is suggested that greater attention be afforded to areas of practice shared by both health care approaches.


The Genetic Basis of Thought Disorder and Language and Communication Disturbances in Schizophrenia.
Thought disorder as well as language and communication disturbances are associated with schizophrenia and are over-represented in clinically unaffected relatives of schizophrenics. All three kinds of dysfunction involve some element of deviant verbalizations, most notably, semantic anomalies. Of particular importance, thought disorder characterized primarily by deviant verbalizations has a higher recurrence in relatives of schizophrenic patients than schizophrenia itself. These findings suggest that deviant verbalizations may be more penetrant expressions of schizophrenia susceptibility genes than schizophrenia. This paper reviews the evidence documenting the presence of thought, language and communication disorders in schizophrenic patients and in their first-degree relatives. This familial aggregation potentially implicates genetic factors in the etiology of thought disorder, language anomalies, and communication disturbances in schizophrenia families. We also present two examples of ways in which thought, language and communication disorders can enrich genetic studies, including those involving schizophrenia.


RNA editing of androgen receptor gene transcripts in prostate cancer cells.
Reactivation of the androgen receptor (AR) signaling pathway represents a critical step in the growth and survival of androgen-independent (AI) prostate cancer (CaP). In this study we show the DU145 and PC3 AI human CaP cell lines respond to androgens and require AR expression for optimal proliferation in vitro. Interestingly, AR gene transcripts in DU145 and PC3 cells harbored a large number of single base pair nucleotide transitions that resulted in missense mutations in selected AR codons. The most notable lesion detected in AR gene transcripts included the oncogenic codon 877T-->A gain-of-function mutation. Surprisingly, AR gene transcript nucleotide transitions were not genome-encoded substitutions, but instead the mutations co-localized to putative A-to-I, U-to-C, C-to-U, and G-to-A RNA editing sites, suggesting the lesions were mediated through RNA editing mechanisms. Higher levels of mRNA encoding the A-to-I RNA editing enzymes ADAR1 and ADARB1 were observed in DU145 and PC3 cells relative to the androgen-responsive LNCaP and 22Rv1 human CaP cell lines, which correlated with higher levels of AR gene transcript A-to-I editing detected in DU145 and PC3 cells. Our results suggest that AR gene transcripts are targeted by different RNA editing enzymes in DU145 and PC3 cells. Thus RNA editing of AR gene transcripts may contribute to the etiology of hormone-refractory phenotypes in advanced stage AI CaP.


Very Long-term Outcome of Bilateral Internal Thoracic Artery in Diabetic Patients: A Systematic Review and Reconstructed Time-To-Event Meta-analysis.
The benefits of single (SITA) and bilateral internal thoracic arteries (BITA) in diabetics undergoing coronary bypass grafting (CABG) are conflicting. We undertook a study-level meta-analysis to compare early and long-term outcomes of both CABG configurations. PubMed, CENTRAL, and EMBASE were searched for studies comparing BITA versus SITA for isolated CABG surgery in diabetics. Randomized trials or observational studies were considered eligible for the analysis. Kaplan-Meier curves of long-term survival were reconstructed and compared with Cox linear regression; incidence rate ratios (IRR) with 95% confidence intervals (CI) for long-term survival were calculated. Landmark analysis and time-varying hazard ratio (HR) were analyzed. Odds ratios (OR) were extracted for early mortality, postoperative stroke, deep sternal wound infection (DSWI), and myocardial infarction (MI). A random effects meta-analysis was performed. Sensitivity analyses included leave-one-out-analyses and meta-regression. Thirteen studies (7332 patients) were included. Overall, at 20-year follow-up, BITA was associated with higher survival (HR = 0.77; 95% CI, 0.71-0.84; P < 0.0001). Time-varying HR and landmark analysis reported BITA was associated with a higher rate of 10-year survival (HR = 0.75, 95% CI 0.68-0.82, P < 0.0001), while from 10 to 20-year follow-up no difference was revealed (HR = 0.99, 95% CI 0.82-1.19, P = 0.93). There was no increase in early mortality, postoperative MI, stroke, or DSWI between the groups. At meta-regression, the higher the age, the higher the long-term overall survival in patients with BITA. In diabetics, the BITA approach is associated with improved 10-year survival with no increase in early mortality, MI, stroke, or DSWI. In the 10-20-year timeframe, BITA and SITA showed comparable survival.


Movement of the velum during speech and singing in classically trained singers.
The present study addresses two questions: (a) Is the action and/or posture of the velopharyngeal valve conducive to allow significant resonance during Western tradition classical singing? (b) How do the actions of the velopharyngeal valve observed in this style of singing compare with normal speech? A photodetector system was used to observe the area function of the velopharyngeal port during speech and classical style singing. Identical speech samples were produced by each subject in a normal speaking voice and then in the low, medium, and high singing ranges. Results indicate that in these four singers the velopharyngeal port was closed significantly longer in singing than in speaking samples. The amount of time the velopharyngeal port was opened was greatest in speech and diminished as the singer ascended in pitch. In the high voice condition, little or no opening of the velopharyngeal port was measured.


Severity-of-Illness Scores and Discharge Disposition in Patients Admitted to Long-Term Acute Care Hospitals.
After an intensive care unit (ICU) admission, nearly 20% of survivors of chronic critical illness require admission to a long-term acute care hospital (LTACH) for continued subspecialty care. The effect of the burden of medical comorbidities on discharge disposition after LTACH admission remains unclear. A retrospective cohort study was performed involving patients with chronic critical illness who were discharged from the medical ICU and admitted to an LTACH between 2016 and 2018. The patients' Acute Physiology and Chronic Health Evaluation II (APACHE II), Sequential Organ Failure Assessment (SOFA), Nutrition Risk in the Critically Ill (NUTRIC), and Charlson Comorbidity Index (CCI) scores at the time of LTACH admission were calculated from electronic medical records. The mean scores on each instrument were compared by discharge disposition. A total of 156 patients were admitted to the LTACH from the medical ICU between 2016 and 2018. They had a mean (SD) age of 61.5 (13.3) years, a mean (SD) body mass index of 28.1 (8.3), a median (IQR) ICU stay of 16.3 (1-108) days, and a median (IQR) LTACH stay of 38.2 (1-227) days. Patients who were discharged home had lower mean (SD) APACHE II (14.6 [5.0] vs 18.2 [5.4], P = .01), SOFA (3.3 [2.1] vs 4.6 [2.1], P = .03), NUTRIC (3.3 [1.4] vs 4.6 [1.4], P = .001), and CCI (4.3 [2.5] vs 6.1 [2.8], P = .02) scores on admission to the LTACH than those who were not discharged home. Severity-of-illness scores on admission to an LTACH can be used to predict patients' likelihood of being discharged home.


A role for the fusogen eff-1 in epidermal stem cell number robustness in Caenorhabditis elegans.
Developmental patterning in Caenorhabditis elegans is known to proceed in a highly stereotypical manner, which raises the question of how developmental robustness is achieved despite the inevitable stochastic noise. We focus here on a population of epidermal cells, the seam cells, which show stem cell-like behaviour and divide symmetrically and asymmetrically over post-embryonic development to generate epidermal and neuronal tissues. We have conducted a mutagenesis screen to identify mutants that introduce phenotypic variability in the normally invariant seam cell population. We report here that a null mutation in the fusogen eff-1 increases seam cell number variability. Using time-lapse microscopy and single molecule fluorescence hybridisation, we find that seam cell division and differentiation patterns are mostly unperturbed in eff-1 mutants, indicating that cell fusion is uncoupled from the cell differentiation programme. Nevertheless, seam cell losses due to the inappropriate differentiation of both daughter cells following division, as well as seam cell gains through symmetric divisions towards the seam cell fate were observed at low frequency. We show that these stochastic errors likely arise through accumulation of defects interrupting the continuity of the seam and changing seam cell shape, highlighting the role of tissue homeostasis in suppressing phenotypic variability during development.


The associations between autistic and communication traits in parents and developmental outcomes in children at familial risk of autism at 6 and 24 months of age.
Several studies have explored relationships between parent broader autism phenotype and offspring communication, and have reported that autistic-like traits in parents are related to offspring communication difficulties and autism severity. However, past research has focused on studying such associations in childhood and we know very little about them in infancy. With accumulating evidence that interventions administered during infancy may be most effective in reducing ASD symptoms, it is imperative to examine whether relationships between parent autistic-like traits and child communication appear even earlier during this critical period of life. This longitudinal study collected data from infant siblings of autistic children (N = 32) and infants with no family history of autism (N = 45) to explore how autistic-like traits in parents related to child developmental outcomes during infancy. Parental communication difficulties and autistic-like traits were found to be associated with a range of child behaviours in the first two years of life, including social-emotional difficulties at 6 and 24 months, lower communication and emerging cognition at 24 months, and increased autistic behaviours at 24 months. Based on the results, it appears that some of the difficulties seen in parents are relayed to children genetically. These findings contribute to ASD research concerning early communication development in children and heritability of ASD traits and may have important implications in monitoring child development. Furthermore, since the current study found a significant association between autistic traits in parents and child social-emotional behaviour as early as 6 months of age, it provides evidence of the value of assessing interventions that target infancy.


Emulsified and Liquid-Liquid Phase-Separated States of α-Pinene Secondary Organic Aerosol Determined Using Aerosol Optical Tweezers.
We demonstrate the first capture and analysis of secondary organic aerosol (SOA) on a droplet suspended in an aerosol optical tweezers (AOT). We examine three initial chemical systems of aqueous NaCl, aqueous glycerol, and squalane at ∼75% relative humidity. For each system we added α-pinene SOA-generated directly in the AOT chamber-to the trapped droplet. The resulting morphology was always observed to be a core of the original droplet phase surrounded by a shell of the added SOA. We also observed a stable emulsion of SOA particles when added to an aqueous NaCl core phase, in addition to the shell of SOA. The persistence of the emulsified SOA particles suspended in the aqueous core suggests that this metastable state may persist for a significant fraction of the aerosol lifecycle for mixed SOA/aqueous particle systems. We conclude that the α-pinene SOA shell creates no major diffusion limitations for water, glycerol, and squalane core phases under humid conditions. These experimental results support the current prompt-partitioning framework used to describe organic aerosol in most atmospheric chemical transport models and highlight the prominence of core-shell morphologies for SOA on a range of core chemical phases.


Tricornered/NDR kinase signaling mediates PINK1-directed mitochondrial quality control and tissue maintenance.
Eukaryotes employ elaborate mitochondrial quality control (MQC) to maintain the function of the power-generating organelle. Parkinson's disease-associated PINK1 and Parkin actively participate in MQC. However, the signaling events involved are largely unknown. Here we show that mechanistic target of rapamycin 2 (mTORC2) and Tricornered (Trc) kinases act downstream from PINK1 to regulate MQC. Trc is phosphorylated in mTORC2-dependent and mTORC2-independent manners and is specifically localized to mitochondria in response to PINK1, which regulates mTORC2 through mitochondrial complex-I activity. Genetically, mTORC2 and Trc act upstream of Parkin. Thus, multiplex kinase signaling is acting between PINK1 and Parkin to regulate MQC, a process highly conserved in mammals.


Physiological acclimation of elk during population restoration in the Missouri Ozarks, USA.
Conservation translocations-the intentional movement of animals to restore populations-have increased over the past 30 years to halt and reverse species declines and losses. However, there are many challenges translocated animals face that should be considered for restoration programs to be successful. Understanding how long it takes for translocated animals to acclimate to these challenges and their new landscape is a critical component of post-release population management. Physiological measures such as hormone responses are increasingly used to assess animal responses and acclimation to disturbances including translocation. We determined the physiological acclimation period of elk (Cervus canadensis) translocated to the Missouri Ozarks, USA, as part of a restoration effort. From 2011 to 2013, we translocated 108 GPS-radio-collared elk from Kentucky, USA, to Missouri, USA, and collected faecal samples for glucocorticoid metabolite extraction to use as an indicator of physiological acclimation. We modelled the response of population-wide faecal glucocorticoid metabolites (fGCMs) across the initial 9 years of the restoration in response to days following release and additional site-specific covariates. Presence of white-tailed deer (Odocoileus virginianus) hunts and monthly precipitation levels were positively and negatively associated with fGCM levels, respectively. Concurrent with influences from site-specific conditions on the release landscape, fGCM levels declined following release. We identified a breakpoint in fGCM decline at ~42 days following translocation releases suggesting elk acclimated physiologically relatively quickly compared to other species. The fast physiological acclimation by Missouri elk suggests effective use of temporary post-release management efforts. Determining how quickly animals acclimate following translocations allows researchers to tailor post-release management plans to each species' needs, thus maximizing the success of future translocation efforts while minimizing costs.


Global RNA modifications to the MALAT1 triple helix differentially affect thermostability and weaken binding to METTL16.
Therapeutic mRNAs are generated using modified nucleotides, namely N1-methylpseudouridine (m1Ψ) triphosphate, so that the mRNA evades detection by the immune system. RNA modifications, even at a single-nucleotide position, perturb RNA structure, although it is not well understood how structure and function is impacted by globally modified RNAs. Therefore, we examined the MALAT1 triple helix, a highly structured stability element that includes single-, double- and triple-stranded RNA, globally modified with N6-methyladenosine (m6A), pseudouridine (Ψ), or m1Ψ. UV thermal denaturation assays showed that m6A destabilizes both the Hoogsteen and Watson-Crick faces of the RNA by ∼20 °C, Ψ stabilizes the Hoogsteen and Watson-Crick faces of the RNA by ∼12 °C, and m1Ψ has minimal effect on the stability of the Hoogsteen face of the RNA but increases the stability of the Watson-Crick face by ∼9 °C. Native gel-shift assays revealed that binding of the methyltransferase-like protein METTL16 to the MALAT1 triple helix was weakened by at least 8-, 99- and 23-fold, respectively, when RNA is globally modified with m6A, Ψ or m1Ψ. These results demonstrate that a more thermostable RNA structure does not lead to tighter RNA-protein interactions, thereby highlighting the regulatory power of RNA modifications by multiple means.


Degradation kinetics of butylmethoxydibenzoylmethane (avobenzone) in aqueous solution.
The degradation kinetics of avobenzone in aqueous solution was studied at 60 +/- 0.2 degrees C over a pH range of 2.0-10.0. The degradation rates were determined by high performance liquid chromatography. The reaction is found to follow first-order kinetics and the rate constant for the decomposition at 25 degrees C is estimated by extrapolation. The breakdown of avobenzone is shown to be hydroxide ion catalyzed and the Arrhenius plots showed the temperature dependence of avobenzone degradation.


CD1c molecules broadly survey the endocytic system.
The ability of antigen-presenting cells to sample distinct intracellular compartments is crucial for microbe detection. Major histocompatibility complex class I and class II molecules sample the cytosol or the late endocytic compartment, allowing detection of microbial peptide antigens that arise in distinct intracellular compartments. In contrast, CD1a and CD1b molecules mediate the presentation of lipid and glycolipid antigens and differentially sample early recycling endosomes or late endocytic compartments, respectively, that contain distinct sets of lipid antigens. Here, we show that, unlike the other CD1 isoforms or major histocompatibility complex molecules that each sample restricted only intracellular compartments, CD1c is remarkable in that it distributes broadly throughout the endocytic system and is expressed in both recycling endosomes and late endocytic compartments. Further, in contrast to CD1b, which requires an acidic environment to function, antigen presentation by CD1c was able to overcome dependence on vesicular acidification. Because CD1c is expressed on essential antigen-presenting cells, such as epidermal Langerhans cells (in the absence of CD1b), or on B cells (without CD1a or -b), we suggest that CD1c molecules allow a comprehensive survey for lipid antigens throughout the endocytic system even in the absence of other CD1 isoforms.


Policy aspects and nursing care of families with parents who are sexual minorities.
Families in which parents are lesbian, gay, bisexual, or transgender are more diverse than they are similar. This article reviews current literature to identify fundamental issues facing families that include sexual minority parents and their children. The unique nursing needs of families with gay, lesbian, transgender, or bisexual parents are critically examined for direct relevance to family nursing practice. Nurses and other health care professionals can incorporate current knowledge of unique child developmental, parenting, and legal issues into their work with these families. Nursing assessment and policies that consider the unique needs of these families will be addressed.


Medical management of acute ocular hypertension in a western screech owl (Megascops kennicottii).
A wild young adult western screech owl (Megascops kennicottii) of unknown sex was presented for evaluation of an abnormal left eye (OS). Ophthalmic examination OS revealed raised intraocular pressure (37 mm Hg; reference interval 7-16 mm Hg), mydriasis, conjunctival and episcleral hyperemia, shallow anterior chamber due to anterior displacement of the lens and iris, rubeosis iridis, and engorgement of the pecten. The intraocular pressure in the right eye (OD) was 11 mm Hg. Multifocal pale, variably translucent, curvilinear to vermiform opacities were observed in the medial and ventral peripheral regions of the retina OD, consistent with focal retinitis. Mannitol (0.46 g/kg IV) was administered over 10 minutes. Forty minutes later, the intraocular pressure was 27 mm Hg OS and 13 mm Hg OD. Dorzolamide (one drop OS q12h), diclofenac (one drop OU q8-12h), and meloxicam (0.5 mg/kg PO q24h) were administered for 3 days. The intraocular pressure OS was within normal limits 1 day (11 mm Hg), 7 days (13 mm Hg), and 4 weeks (14 mm Hg) after this treatment. Complications arising during hospitalization and rehabilitation included superficial corneal ulceration of both eyes presumed secondary to trauma on being caught and superficial damage to a talon. The owl was released after a period of rehabilitation. Characteristic presenting signs as well as response to therapy suggest aqueous misdirection was the cause of ocular hypertension in this owl. To our knowledge, this is the first report of suspected aqueous misdirection and its medical management in a raptor.


Violence, sexual abuse and health in Greenland.
The purposes of the study were to analyse the lifetime prevalence of violence and sexual abuse among the Inuit in Greenland and to study the associations between health and having been the victim of violence or sexual abuse. Associations were studied with specific attention to possible differences between women and men. Further, response rates were analysed specifically in order to understand consequences of including questions on violence and sexual abuse in the questionnaire survey. The analyses were based on material from a cross-sectional health interview survey conducted during 1993-94 with participation from a random sample of the Inuit population in Greenland (N = 1393). The prevalence of ever having been a victim of violence was 47% among women and 48% among men. Women had more often than men been sexually abused (25% and 6%) (p < 0,001) and had more often been sexually abused in childhood (8% and 3%) (p = 0.001). Having been the victim of violence or sexual abuse was significantly associated with a number of health problems: chronic disease, recent illness, poor self-rated health, and mental health problems. The associations between having been the victim of violence or sexual abuse and health was stronger for women than for men. It is possible to secure a reasonably high response rate in a general health survey that includes questions on violence and sexual abuse.


Dental health knowledge in a group of Black adolescents living in rural South Carolina.
This study investigated levels of dental health knowledge and factors associated with adequate dental health knowledge in a group of black adolescents living in rural areas. Using a self-administered questionnaire, data were collected on a convenience sample of 151 black adolescents aged 10 to 18 yrs living in rural South Carolina. The mean percent correct on the dental health knowledge questions was 55.0%. Using 75% as the cutoff for adequate dental health knowledge, only 7.9% of the respondents achieved this level. Two thirds of the younger adolescents (aged 10-12 yrs) were below the median on dental health knowledge. Respondents selected dental health professionals, family, and school as the three main sources of dental health information. Results from univariate logistic regression analyses indicated that being older, having a regular dentist for routine care, and having learned about dental health from dental health professionals, family, mass media, and friends were significantly associated with adequate dental health knowledge. After adjusting for other explanatory factors, adequate dental health knowledge was associated with being older and having learned about dental health from friends. This group of adolescents seems to have limited dental health knowledge with misconceptions concerning periodontal health and caries prevention. This was especially evident in younger adolescents. Incorporation of peer dental health education in school is worthwhile to investigate as a means to enhance the dental health knowledge of these adolescents.


Evaluation of High-Pitch Ungated Pediatric Cardiovascular Computed Tomography for the Assessment of Cardiac Structures in Neonates.
We evaluated a high-pitch, non-electrocardiogram-gated cardiac computed tomographic protocol, designed to image both cardiac and extracardiac structures, including coronary arteries, in a neonatal population (less than 1 year old) that was referred for congenital heart disease assessment and compared it with an optimized standard-pitch protocol in an equivalent cohort. Twenty-nine high-pitch scans were compared with 31 age-matched, sex-matched, and weight-matched standard-pitch, dosimetrically equivalent scans. The visualization and subjective quality of both cardiac and extracardiac structures were scored by consensus between 2 trained blinded observers. Image noise, signal-to-noise and contrast-to-noise ratios, and radiation doses were also compared. The high-pitch protocol better demonstrated the pulmonary veins (P=0.03) and all coronary segments (all P<0.05), except the distal right coronary artery (P=0.10), with no significant difference in the visualization of the remaining cardiac or extracardiac structures. Both contrast-to-noise and signal-to-noise ratios improved due to greater vessel opacity, with significantly fewer streak (P<0.01) and motion (P<0.01) artifacts. Image noise and computed tomographic dose index were comparable across the 2 techniques; however, the high-pitch acquisition resulted in a small, but statistically significant, increase in dose-length product [13.0 mGy.cm (9.0 to 17.3) vs. 11.0 mGy.cm (9.0 to 13.0), P=0.05] due to greater z-overscanning. In neonates, a high-pitch protocol improves coronary artery and pulmonary vein delineation compared with the standard-pitch protocol, allowing a more comprehensive assessment of cardiovascular anatomy while obviating the need for either patient sedation or heart rate control.


Health and income inequalities in Europe: What is the role of circumstances?
Equality of opportunity theories distinguish between inequalities due to individual effort and those due to external circumstances. Recent research has shown that half of the variability in income of World population was determined by country of birth and income distribution. Since health and income are generally strictly related, the aim of this paper is to estimate how much variability in income and health is determined by external circumstances. We use data from the Survey of Health, Ageing and Retirement (SHARE) and the English Longitudinal Survey on Ageing (ELSA), two comparable multidisciplinary surveys that provide micro-level data on health and financial resources among the elderly for a large number of European countries. Our baseline estimation shows that about 20% of the variability in income is explained by current country-specific circumstances, while health outcomes range from 12% using BMI to 19% using self-rated health. By including early-life circumstances, the explained variability increases almost 20 percentage points for income and for self-rated health but less for other health outcomes. Finally, by controlling for endogeneity issues linked with effort, our estimates indicate that circumstances better explain variability in health outcomes. Results are robust to some tests, and the implications of these findings are discussed.


[Ostearticular tuberculosis. Study of 53 cases].
To analyze the characteristics of bone and joint tuberculosis in a Spanish university hospital. Retrospective study (1984-2006) in a university hospital. All patients had microbiologically and/or histologically proved osteoarticular tuberculosis. Patients with clinical and radiological criteriae, positive tuberculin test responding to antituberculous treatment were not excluded despite negative cultures. We attended 53 patients with osteoarticular tuberculosis (35 males/18 females; mean age: 52 years). Bone tuberculosis involved axial skeleton in 37 patients (71%), peripheral distribution in 12 (21%) and both locations in 4 (8%). Mean time to diagnosis was 8 months. Most common involved joints were knee and ankle. An extraarticular involvement was found in 12 patients (22%) and multifocal bone infection in 6 (11%). Risk factors were present in 22 patients (42%), and 15% were immigrants. The diagnosis was established by a positive culture in 40 cases (75%), and 33 (62%) had suggestive histology. Complications of tuberculosis included medullar compression (9.4%), abscess (12%) and fistulae (9.4%). Spine involvement required surgical intervention in 27% and peripheral involvement in 56%. Curation was achieved in 33 patients (62.3%), curation with secuelae in 19 cases (35.9%) and no consolidation of arthrodesis in one case. Bone and joint tuberculosis is still common in our area and should be particularly considered in immigrants. Diagnosis delay is remarkable. Complications are present in one third of patients. Surgical treatment is often required.


Hepatic drug metabolism and lipid peroxidation in thiamine deficient rats.
In vitro metabolism of aminopyrene, ethylmorphine (Type I substrates), N-methylaniline and acetanilide (Type II substrates) in liver microsomal fraction from thiamine deficient male and female rats was studied. No significant change in microsomal protein content was noticed during the period of thiamine deficiency. However, a significant increase in the in vitro oxidation of aminopyrene, ethylmorphine, N-methylaniline and hydroxylation of acetanilide was observed. The NADPH linked and ascorbate induced lipid peroxidation was also increased during thiamine deficiency. The levels of NADPH cytochrome c-reductase, cytochrome b5 and heme were noticeably increased in thiamine deficient animals as compared to normal rats. Phenobarbital treatment induced the activities of all drug enzymes and inhibited the lipid peroxidation in either sex during the period of thiamine deficiency. It appears that thiamine intake is an important determination in drug metabolism and lipid peroxidation.


Pediatric Hypertension.
The prevalence of elevated blood pressure and hypertension in children and adolescents has increased over the past decade. This trend is most likely related to increases in primary hypertension associated with increasing obesity rates in children. Lifestyle as well as genetics play a significant role in the development of primary hypertension. Hypertension in children and adolescents is under-recognized and undertreated. The 2017 Revised Clinical Practice Guideline for Screening and Management of High Blood Pressure in Children and Adolescents aimed to create new normative blood pressure tables using data from healthy weight children, meaning between the 5th and 85th percentile per the Centers for Disease Control and Prevention. Another important objective was to simplify normative data to ease screening and detection of elevated blood pressures. The consequences of chronic hypertension are significant, with its major affect being in poor cardiovascular health outcomes both in childhood and early adulthood. Challenges to detection and adequate treatment should be overcome with continued education and awareness to prevent the long-term effects of uncontrolled hypertension that starts in childhood. [Pediatr Ann. 2018;47(12):e499-e503.].


[Management of chronic pain after inguinal hernioplasty].
Chronic groin pain after herniorrhaphy is a concern, as approximately 10% of patients undergoing inguinal hernia repair have symptoms, which often limit physical ability. The etiopathogenesis is related to periostitis pubis (somatic pain) and more often to nerve injury (neuropathic pain). It is clinically important to distinguish between these two types of pain because treatment can be different. The physician should establish a routine diagnosis and treatment, and most patients will need surgical approach. Prevention of this condition is of great importance and can lead to a lower incidence of the syndrome. Some measures are key, such as how to avoid application of stitches or clips to the pubis periosteum, using the prosthesis carefully and identifying the nerves in the groin. This last measure is certainly the most important in the prevention of chronic pain and involves thorough knowledge of anatomy and the use of refined technique.


Plasma matrix metalloproteinase-9 and left ventricular remodelling after acute myocardial infarction in man: a prospective cohort study.
To describe temporal profiles of plasma matrix metalloproteinases (MMP-2 and MMP-9), and their relationship with echocardiographic (Echo) parameters of left ventricular (LV) function and remodelling, after acute myocardial infarction (AMI) in man. Plasma MMP-2 and MMP-9 were assayed at intervals (0-12, 12-24, 24-48, 48-72, 72-96, and > 96 h) in 91 patients with AMI (ST-elevation/non-ST-elevation 77/24; 73% male; 40% anterior site) and on a single occasion in 172 age- and sex-matched control subjects with stable coronary artery disease. Echo assessment of LV volumes, LV ejection fraction (LVEF), and wall motion index score were assessed before discharge and at follow-up (median 176, range 138-262 days) for patients and on a single occassion in controls. Plasma MMP-2 was similar at all times after AMI, elevated when compared with control (P = 0.005-0.001) and unrelated to LV function or volume during index admission or at follow-up. Maximal MMP-9 was seen at 0-12 h and was elevated when compared with control (P = 0.002) followed by fall to a plateau. Both maximal and plateau MMP-9 concentration correlated with white blood cell (WBC, P = 0.023 to < 0.001) and neutrophil count (P = 0.014 to < 0.001). Maximal MMP-9 had independent predictive value for lower LVEF (P = 0.004) during admission and for greater change in LV end-diastolic volume between admission and follow-up (R = 0.3, P = 0.016). In contrast, higher plateau levels of MMP-9 were associated with relative preservation of LV function (increasing LVEF, P = 0.002; decreasing WMIS, P = 0.009) and less change in end-systolic volume and end-diastolic volumes after discharge (P = 0.001 and 0.024, respectively). Both MMP-9 and MMP-2 are elevated following AMI. The biphasic profile of plasma MMP-9 is related to LV remodelling and function following AMI in man. Higher early levels of MMP-9 associate with the extent of LV remodelling and circulating WBC levels. In contrast, higher plateau levels later after AMI are associated with relative preservation of LV function. Temporal profile, rather than absolute magnitude, of MMP-9 activity appears to be important for LV remodelling after AMI.


Catechin is present as metabolites in human plasma after consumption of red wine.
Flavonoids are components of fruits, vegetables and wines. An abundance of flavonoids in the diet is correlated with reduced heart disease mortality, suggesting that they act as protective nutrients. However, little is known about the absorption and metabolism of flavonoids after normal foods are consumed. This study measured the levels of one abundant flavonoid, (+)-catechin, and its metabolites in plasma after five male and four female volunteers consumed 120 mL of red wine (RW) one day and de-alcoholized red wine (DRW) on a separate day. Each wine sample contained 35 +/- 1 mg catechin (mean +/- SEM). Plasma levels of catechin and its metabolite 3'-O-methylcatechin (3'MC) were determined by gas chromatography-mass spectrometry (GC-MS) of the trimethylsilylated (TMS) derivatives. Glucuronide and sulfate conjugates were determined after enzymatic hydrolysis. Before RW or DRW consumption, plasma levels of catechin, 3'MC and all conjugates were <2 nmol/L. After 1 h, average levels of catechin, 3'MC and all conjugates increased to 91 +/- 14 nmol/L (RW) and 81 +/- 11 nmol/L (DRW). At 1 h, 21 +/- 1% of the metabolites were methylated and <2% of catechin and 3'MC were unconjugated. Catechin was present as both a sulfate conjugate and a conjugate containing both glucuronide and sulfate residues. 3'MC was present primarily as a glucuronide conjugate. At every time point, catechin was present almost exclusively as metabolites, and these levels were independent of ethanol. Therefore, if flavonoids are protective nutrients, the active forms are likely to be metabolites, which are far more abundant in plasma than the forms that exist in foods.


Detection of Abacavir-Induced Structural Alterations in Human Leukocyte Antigen-B*57 : 01 Using Phage Display.
The interaction of human leukocyte antigen (HLA) with specific drugs is associated with delayed-type hypersensitivity reactions, which cause severe cutaneous toxicity. Such interactions induce structural alterations in HLA complexes via several different mechanisms such as the hapten theory, p-i concept, and altered peptide repertoire model, leading to the activation of cytotoxic T cells. To date, comprehensive detection of such structural alterations in preclinical studies has been difficult. Here, we evaluated structural alterations in HLA complexes focusing on the interaction between the HLA-B*57 : 01 allele and abacavir (an anti-human immunodeficiency virus drug), representing a model of abacavir hypersensitivity syndrome induced by changes in the peptide repertoire on the HLA molecule. We employed a phage display method using a commercially available antibody library to screen specific phage antibodies able to recognize HLA-B*57 : 01. The affinity of selected phage antibodies increased because of structural alterations in HLA-B*57 : 01 following exposure to abacavir, indicating that specific phage antibodies can identify drug-mediated structural changes in HLA complexes. We also identified an unreported structural change in HLA-B*57 : 01 using the phage display method, whereby abacavir increased the expression of peptide-deficient HLA-B*57 : 01 on the cell surface. These results suggest that phage display technology is a useful method for detecting structural changes in HLA complexes. This technology represents a potential novel strategy for predicting HLA-associated hypersensitivity reactions by drugs in pre-clinical studies.


Identification of flowering genes in Camellia perpetua by comparative transcriptome analysis.
Camellia perpetua has the excellent characteristic of flowering multiple times throughout the year, which is of great importance to solve the problem of "short flowering period" and "low fresh flower yield" in the yellow Camellia industry at present. Observations of flowering phenology have demonstrated that most floral buds of C. perpetua were formed by the differentiation of axillary buds in the scales at the base of the terminal buds of annual branches. However, the molecular mechanism of flowering in C. perpetua is still unclear. In this study, we conducted a comparative transcriptomic study of the terminal buds and their basal flower buds in March (spring) and September (autumn) using RNA-seq and found that a total of 11,067 genes were significantly differentially expressed in these two periods. We identified 27 genes related to gibberellin acid (GA) synthesis, catabolism, and signal transduction during floral bud differentiation. However, treatment of the terminal buds and axillary buds of C. perpetua on annual branch with GA3 did not induce floral buds at the reproductive growth season (in August) but promoted shoot sprouting. Moreover, 203 flowering genes were identified from the C. perpetua transcriptome library through homology alignment, including flowering integrators LEAFY (LFY) and UNUSUAL FLORAL ORGANS (UFO), as well as MADS-box, SQUAMOSA PROMOTER BINDING PROTEIN-box (SBP-box), and TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTOR (TCP) genes, which were specifically upregulated in floral buds and were likely involved in flowering in C. perpetua. The floral inhibitor CperTFL1b was identified and cloned from C. perpetua, and its expression level was specifically regulated in terminal buds in autumn. Ectopic overexpression of CperTFL1b delayed flowering time and produced abnormal inflorescence and floral organs in Arabidopsis, suggesting that CperTFL1b inhibits flowering. In conclusion, this study deepens our understanding of the molecular mechanism of blooms throughout the year in C. perpetua and provides a helpful reference for cultivating new varieties of yellow Camellia with improved flowering traits.


Biological and Molecular Effects of Trypanosoma cruzi Residence in a LAMP-Deficient Intracellular Environment.
Trypanosoma cruzi invades non-professional phagocytic cells by subverting their membrane repair process, which is dependent on membrane injury and cell signaling, intracellular calcium increase, and lysosome recruitment. Cells lacking lysosome-associated membrane proteins 1 and 2 (LAMP1 and LAMP2) are less permissive to parasite invasion but more prone to parasite intracellular multiplication. Several passages through a different intracellular environment can significantly change T. cruzi's gene expression profile. Here, we evaluated whether one single passage through LAMP-deficient (KO) or wild-type (WT) fibroblasts, thus different intracellular environments, could influence T. cruzi Y strain trypomastigotes' ability to invade L6 myoblasts and WT fibroblasts host cells. Parasites released from LAMP2 KO cells (TcY-L2-/-) showed higher invasion, calcium signaling, and membrane injury rates, for the assays in L6 myoblasts, when compared to those released from WT (TcY-WT) or LAMP1/2 KO cells (TcY-L1/2-/-). On the other hand, TcY-L1/2-/- showed higher invasion, calcium signaling, and cell membrane injury rates, for the assays in WT fibroblasts, compared to TcY-WT and TcY-L1/2-/-. Albeit TcY-WT presented an intermediary invasion and calcium signaling rates, compared to the others, in WT fibroblasts, they induced lower levels of injury, which reinforces that signals mediated by surface membrane protein interactions also have a significant contribution to trigger host cell calcium signals. These results clearly show that parasites released from WT or LAMP KO cells are distinct from each other. Additionally, these parasites' ability to invade the cell may be distinct depending on which cell type they interact with. Since these alterations most likely would reflect differences among parasite surface molecules, we also evaluated their proteome. We identified few protein complexes, membrane, and secreted proteins regulated in our dataset. Among those are some members of MASP, mucins, trans-sialidases, and gp63 proteins family, which are known to play an important role during parasite infection and could correlate to TcY-WT, TcY-L1/2-/-, and TcY-L2-/- biological behavior.


Phase I and Biomarker Study of Plerixafor and Bevacizumab in Recurrent High-Grade Glioma.
Purpose: Although antiangiogenic therapy for high-grade glioma (HGG) is promising, responses are not durable. Correlative clinical studies suggest that the SDF-1α/CXCR4 axis may mediate resistance to VEGFR inhibition. Preclinical data have demonstrated that plerixafor (a reversible CXCR4 inhibitor) could inhibit glioma progression after anti-VEGF pathway inhibition. We conducted a phase I study to determine the safety of plerixafor and bevacizumab in recurrent HGG.Patients and Methods: Part 1 enrolled 23 patients with a 3 × 3 dose escalation design to a maximum planned dose of plerixafor 320 μg/kg subcutaneously on days 1 to 21 and bevacizumab 10 mg/kg intravenously on days 1 and 15 of each 28-day cycle. Cerebrospinal fluid (CSF) and plasma samples were obtained for pharmacokinetic analyses. Plasma and cellular biomarkers were evaluated before and after treatment. Part 2 enrolled 3 patients and was a surgical study to determine plerixafor's penetration in tumor tissue.Results: In Part 1, no dose-limiting toxicities were seen at the maximum planned dose of plerixafor + bevacizumab. Treatment was well tolerated. After plerixafor 320 μg/kg treatment, the average CSF drug concentration was 26.8 ± 19.6 ng/mL. Plerixafor concentration in resected tumor tissue from patients pretreated with plerixafor was 10 to 12 μg/g. Circulating biomarker data indicated that plerixafor + bevacizumab induces rapid and persistent increases in plasma SDF-1α and placental growth factor. Progression-free survival correlated with pretreatment plasma soluble mesenchymal-epithelial transition receptor and sVEGFR1, and overall survival with the change during treatment in CD34+ progenitor/stem cells and CD8 T cells.Conclusions: Plerixafor + bevacizumab was well tolerated in HGG patients. Plerixafor distributed to both the CSF and brain tumor tissue, and treatment was associated with biomarker changes consistent with VEGF and CXCR4 inhibition. Clin Cancer Res; 24(19); 4643-9. ©2018 AACR.


Development of a Corneal and Eye Protection Strategy in Domestic Swine.
Large animal models are essential to research in facial paralysis, face transplant, craniofacial surgery, and ophthalmology. Pigs are a well-studied species with high similarity to human anatomy and physiology for these research areas. However, in contrast to cats and dogs protecting the cornea and eye is difficult in swine due to the inability to use an Elizabethan collar (E-collar) and the complexity of placing and maintaining a temporary tarsorrhaphy for corneal protection due to the strength of the pig levator muscle. This study presents an effective method to provide corneal and eye protection in the domestic swine for at least 50 d. Furthermore, protection of the eye and face is achieved through the innovative use of a modified ophthalmologic face shield. The findings from this study will advance large animal research in these fields, enabling innovation in surgery and tissue engineering in areas of both craniofacial and ophthalmologic research.


Design and Development of a Miniaturized Percutaneously Deployable Wireless Left Ventricular Assist Device: Early Prototypes and Feasibility Testing.
The current left ventricular assist devices (LVADs) are limited by a highly invasive implantation procedure in a severely unstable group of advanced heart failure patients. Additionally, the current transcutaneous power drive line acts as a nidus for infection resulting in significant morbidity and mortality. In an effort to decrease this invasiveness and eliminate drive line complications, we have conceived a wireless miniaturized percutaneous LVAD, capable of being delivered endovascularly with a tether-free operation. The system obviates the need for a transcutaneous fluid purge line required in existing temporary devices by utilizing an incorporated magnetically coupled impeller for a complete seal. The objective of this article was to demonstrate early development and proof-of-concept feasibility testing to serve as the groundwork for future formalized device development. Five early prototypes were designed and constructed to iteratively minimize the pump size and improve fluid dynamic performance. Various magnetic coupling configurations were tested. Using SolidWorks and ANSYS software for modeling and simulation, several geometric parameters were varied. HQ curves were constructed from preliminary in vitro testing to characterize the pump performance. Bench top tests showed no-slip magnetic coupling of the impeller to the driveshaft up to the current limit of the motor. The pump power requirements were tested in vitro and were within the appropriate range for powering via a wireless energy transfer system. Our results demonstrate the proof-of-concept feasibility of a novel endovascular cardiac assist device with the potential to eventually offer patients an untethered, minimally invasive support.


Haematemesis and Melaena: surgical management.
A prospective study of the surgical management of 100 consecutive patients with benign, non-variceal upper gastrointestinal bleeding is presented. The manner of presentation, precipitating factors, investigations and associated medical problems are discussed. Chronic duodenal ulceration was the most common cause of haemorrhage. Vagotomy and drainage with oversewing or excision of the bleeding ulcer was the surgical procedure performed in 71 of the patients in the series. The incidence of recurrent bleeding was 7%; no patient in this category required further operation. The mortality rate was 6%, and there were no deaths recorded in the patients who underwent vagotomy and drainage. The reasons for the relatively low surgical mortality are discussed.