TAD evolutionary and functional characterization reveals diversity in mammalian TAD boundary properties and function.
Topological associating domains (TADs) are self-interacting genomic units crucial for shaping gene regulation patterns. Despite their importance, the extent of their evolutionary conservation and its functional implications remain largely unknown. In this study, we generate Hi-C and ChIP-seq data and compare TAD organization across four primate and four rodent species and characterize the genetic and epigenetic properties of TAD boundaries in correspondence to their evolutionary conservation. We find 14% of all human TAD boundaries to be shared among all eight species (ultraconserved), while 15% are human-specific. Ultraconserved TAD boundaries have stronger insulation strength, CTCF binding, and enrichment of older retrotransposons compared to species-specific boundaries. CRISPR-Cas9 knockouts of an ultraconserved boundary in a mouse model lead to tissue-specific gene expression changes and morphological phenotypes. Deletion of a human-specific boundary near the autism-related AUTS2 gene results in the upregulation of this gene in neurons. Overall, our study provides pertinent TAD boundary evolutionary conservation annotations and showcases the functional importance of TAD evolution.


What's New in Pediatric Orthopaedic Trauma: The Upper Extremity.
Upper extremity fractures are the most common fractures in children. Many high-quality studies have been reported regarding operative and nonoperative treatment of different upper extremity fractures in children. This review will summarize the literature on 4 major upper extremity fractures in children over the past 5 years. PubMed and Embase databases were queried for publications in the English language on supracondylar humerus (SCH) fractures, lateral humeral condyle fractures, medial epicondyle fractures, and clavicle fractures from January 1, 2013 until November 1, 2018. Papers believed to yield significant findings to our profession were included in this review. A total of 1150 studies were related to the search terms, and after cursory assessment, the authors elected to fully review 30 papers for this publication: 12 related to SCH fractures, 10 related to lateral condyle humerus fractures, 3 related to medial epicondyle humerus fractures, and 5 involving clavicle fractures. The level of evidence for these studies was either level III or IV. SCH fractures are increasingly being treated at trauma centers or pediatric hospitals in the United States. The rate of open reduction in this fracture type is decreasing overall, but the flexion type SCH fractures (especially in the setting of ulnar nerve injury) continue to be at increased risk of requiring open reduction. There has been a paradigm shift in the treatment of lateral condyle humerus fractures, wherein authors have demonstrated successful management with closed reduction and percutaneous pinning when an adjunct arthrogram is performed demonstrating articular congruity. More studies are needed to find the optimal treatment for displaced medial epicondyle and clavicle fractures in adolescents, as results to date do not necessarily mirror those seen in the adult population. Level IV.


Nd3+,Ho3+-Codoped apatite-related NaLa9(GeO4)6O2 phosphors for the near- and middle-infrared region.
The apatite-like NaLa9(GeO4)6O2:Nd3+,Ho3+ phosphor is prepared using the solid-state method. Rietveld refinement of high-resolution time-of-flight neutron powder diffraction measurements indicate that this compound crystallizes in the hexagonal system with space group P63/m, Z = 1 and unit cell parameters a = 9.88903(6) Å, c = 7.25602(5) Å, V = 614.521(7) Å3 at room temperature. The 4f sites are statistically occupied by La, Nd and Na, while 6h sites are occupied by La and Nd. Luminescence in the near- and middle-IR range caused by the transitions in neodymium and holmium ions is excited under 808 nm laser diode radiation. The highest emission intensity in NaLa9-x-yNdxHoy(GeO4)6O2 is attained at trace amounts of holmium, and it decreases sharply when y increases to 0.01. The IR phosphors have a good thermal stability and exhibit a very weak upconversion emission in the red spectral range upon 808 nm excitation. A scheme of excitation and emission pathways involving ground/excited state absorption, energy transfer, cross-relaxation, nonradiative multiphonon relaxation processes in Nd3+ and Ho3+ ions has been proposed. The data analysis indicates that Nd3+ ions serve as sensitizers for Ho3+ ions in these compounds, stimulating intense 2.1 μm and 2.7 μm emissions. These apatite-related germanate phosphors are promising materials for near- and middle-infrared solid-state lighting applications.


Phase I and II trial on infusional 5-fluorouracil and gefitinib in combination with preoperative radiotherapy in rectal cancer: 10-years median follow-up.
The aim of this study is to evaluate the long term survival of the addition of gefitinib to chemoradiotherapy (CRT) in locally advanced rectal cancer (LARC). This previously published multicentre, open-label, phase I-II study, enrolled patients (pts) with LARC to receive CRT with concurrent 5-fluorouracil continuous intravenous infusion and a dose escalation of orally administered gefitinib, followed 6-8 weeks later by surgery. An intra-operative radiotherapy boost of 10 Gy was planned. Adjuvant chemotherapy was administrated in ypN1-2 pts. After a median f/u of >10 years, we analyzed Local Control (LC), Metastasis Free Survival (MFS), Disease Free Survival (DFS), Disease Specific Survival (DSS) and Overall Survival (OS). Predictive endpoints of clinical outcomes were tested by univariate and multivariate analysis. Variables analyzed included: age, gefitinib dose and interruptions, adjuvant CT, surgery type, ypT, ypN, and TRG grade. We have also analyzed late toxicity according to CTCAEv4. Of the 41 initially enrolled pts, 39 were evaluable (27M, 12F). With a median f/u of 133 months, LC, MFS, DFS, OS and DSS at 5 years were 84%; 71%; 64%; 87% and 92%, respectively. The OS and DSS at 10 years were 61,5% and 76%, respectively. Grade 3-4 late toxicity occurred in 38% of pts: sexual (28,2%) and gastrointestinal toxicities (10,2%). Long term outcomes and late toxicity were similar to previously reported series. The addition of gefitinib did not improve outcomes in LARC. Gefitinib is not recommended for rectal cancer patients who received 5-FU based preoperative CRT. Further studies may identify if gefitinib is beneficial in selected group of patients.


Treatment of plasma cell myeloma with cytotoxic agents.
Because cross-resistance between alkylating agents has not been observed, we attempt in a prospective trial to determine the advantages, if any, in administering three alkylating agents sequentially, alternately, or concurrently. A patient with myeloma, showing progressive shortening of M-protein doubling time from 98 to 15 days, developed an acute terminal phase, characterized by fever and pancytopenia. A similar acute terminal phase was observed in 17 of 50 deaths from myeloma. Since alkylating agents are only effective in controlling the chronic phase of myeloma, future improvements will require the discovery of agents that delay, prevent, or are effective in the treatment of the acute phase. Forty-five patients with kappa- and 36 with lambda-light-chain disease showed no differences in frequency of amyloidosis, renal failure, response to treatment, or survival after treatment with alkylating agents.


Subthalamic, not striatal, activity correlates with basal ganglia downstream activity in normal and parkinsonian monkeys.
The striatum and the subthalamic nucleus (STN) constitute the input stage of the basal ganglia (BG) network and together innervate BG downstream structures using GABA and glutamate, respectively. Comparison of the neuronal activity in BG input and downstream structures reveals that subthalamic, not striatal, activity fluctuations correlate with modulations in the increase/decrease discharge balance of BG downstream neurons during temporal discounting classical condition task. After induction of parkinsonism with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), abnormal low beta (8-15 Hz) spiking and local field potential (LFP) oscillations resonate across the BG network. Nevertheless, LFP beta oscillations entrain spiking activity of STN, striatal cholinergic interneurons and BG downstream structures, but do not entrain spiking activity of striatal projection neurons. Our results highlight the pivotal role of STN divergent projections in BG physiology and pathophysiology and may explain why STN is such an effective site for invasive treatment of advanced Parkinson's disease and other BG-related disorders.


Misincorporation during DNA synthesis, analyzed by gel electrophoresis.
A method has been developed for simultaneous comparison of the propensity of a DNA polymerase to misincorporate at different points on a natural template-primer. In this method elongation of a [5'-32P] primer, annealed to a bacteriophage template strand, is carried out in the presence of only three dNTPs (highly purified by HPLC). Under these conditions the rate of primer elongation (monitored by gel electrophoresis/autoradiography) is limited by the rate of misincorporation at template positions complementary to the missing dNTP. Variations in the rate of elongation (revealed by autoradiographic banding patterns) reflect variations in the propensity for misincorporation at different positions along the template. The effect on primer elongation produced by addition of a chemically modified dNTP to 'minus' reactions reveals the mispairing potential of the modified nucleotide during DNA synthesis. By use of this electrophoretic assay of misincorporation we have demonstrated that the fidelity of E. coli DNA polymerase I varies greatly at different positions along a natural template, and that BrdUTP and IodUTP can be incorporated in place of dCTP during chain elongation catalyzed by this enzyme.


New sensitive method for the measurement of lysozyme and lactoferrin to explore mucosal innate immunity. Part II: time-resolved immunofluorometric assay used in HIV patients with oral candidiasis.
The aim of this study was to explore lysozyme and lactoferrin concentrations in human immunodeficiency virus (HIV)-infected patients with oropharyngeal candidiasis (OPC). These proteins were measured by time-resolved immunofluorometric assay, validated in Part I of this study, in paired serum and salivary secretions of 30 patients. Eleven HIV-positive patients without OPC, eight HIV-positive patients with OPC and eleven HIV-negative healthy subjects were included in the study. The relative coefficient of excretion of salivary albumin was used to establish protein origin. In serum, the low lactoferrin concentrations in HIV-infected patients with and without OPC (0.610 mg/l (p < 0.05) and 0.896 mg/l (p < 0.01) vs. 1.439 mg/l in healthy subjects) were probably due to a decrease in nonspecific immunity, particularly the polymorphonuclear cells. In HIV-infected patients with OPC, the high salivary lysozyme and lactoferrin concentrations (170.94 mg/l and 66.48 mg/l vs. 23.35 mg/l and 10.20 mg/l in healthy subjects, respectively) and their mean relative coefficient of excretion of above 1 indicated a high local production of lysozyme and lactoferrin in saliva. The development of OPC in HIV-infected patients could be a consequence of inefficient lysozyme and lactoferrin concentrations and of decreased cooperation between innate and adaptative immune systems.


Aerodynamics and mathematics in National Socialist Germany and Fascist Italy: a comparison of research institutes.
The article is concerned with the mathematical sciences in National Socialist Germany and Fascist Italy, with special attention to research important to the war effort. It focuses on three institutional developments: the expansion of the Kaiser Wilhelm Institute for Fluid Dynamics in Göttingen, the foundation of the Reich Institute for Mathematics in Oberwolfach (Black Forest), and the work of the Istituto Nazionale per le Applicazioni del Calcolo in Rome. All three developments are embedded in the general political background, thus providing a basis for comparative conclusions about the conditions of the mathematical sciences and military-related research in Germany and Italy. It turns out that in both countries, the increasing demand for mathematical knowledge in modern warfare led to the establishment of "extra-university" national institutions specifically devoted to mathematical research.


Randomized, double-blind, placebo-controlled, international trial of the oral IIb/IIIa antagonist lotrafiban in coronary and cerebrovascular disease.
This is the primary report of the large-scale evaluation of lotrafiban, an orally administered IIb/IIIa receptor antagonist, a unique trial with respect to the platelet antagonist, protocol design, and inclusion of cerebrovascular disease in a significant proportion of patients. Patients with vascular disease were randomized to lotrafiban 30 or 50 mg BID on the basis of age and predicted creatinine clearance or placebo in addition to aspirin at a dose ranging from 75 to 325 mg/d at the discretion of the physician-investigator. Follow-up was for up to 2 years. The primary end point was the composite of all-cause mortality, myocardial infarction, stroke, recurrent ischemia requiring hospitalization, and urgent revascularization. Of 9190 patients enrolled from 23 countries and 690 hospitals, 41% had cerebrovascular disease at the time of entry, and 59% had coronary artery disease. Death occurred in 2.3% of placebo-assigned patients and 3.0% of lotrafiban-group patients (hazard ratio 1.33, 95% CI 1.03 to 1.72, P=0.026), and the cause of excess death was vascular related. There was no significant difference in the primary end point (17.5% compared with 16.4%, respectively; hazard ratio 0.94, 95% CI 0.85 to 1.03, P=0.19). Serious bleeding was more frequent in the lotrafiban group (8.0% compared with 2.8%; P<0.001). Serious bleeding was more common among patients who received higher doses of aspirin (>162 mg/d), with or without lotrafiban. Lotrafiban, an orally administered platelet glycoprotein IIb/IIIa blocker, induced a 33% increase in death rate, which was vascular in origin and not affected by the type of atherosclerotic involvement at entry to the trial. Although the dose of aspirin was not randomly assigned, the finding of increased bleeding with doses >162 mg/d is noteworthy.


Myocardial perfusion and metabolism after transcoronary ablation of septum hypertrophy (TASH) in hypertrophic obstructive cardiomyopathy.
Outflow obstruction constitutes a major problem in hypertrophic obstructive cardiomyopathy (HOCM) and may be treated by transcoronary injection of ethanol into septal arteries (transcoronary ablation of septum hypertrophy, TASH). We report on myocardial perfusion and glucose metabolism in a 50 year old man in whom TASH led to a reduction of septal thickness (9 vs. 26 mm), a sustained elimination of the outflow tract obstruction (resting gradient 0 vs. 60 mmHg; post-ES-gradient 10 vs. 145 mmHg) and a substantial clinical improvement (NYHA stage II vs. II-III) without impairment of global ventricular function (left ventricular ejection fraction 0.62 vs. 0.64). After TASH, perfusion and glucose metabolism were assessed by positron emission tomography (PET) using F-18-fluorodeoxyglucose (F-18-FDG) and Tc-99m-MIBI single photon emission tomography (SPECT). TASH results in matched reduction of perfusion and glucose consumption in a circumscribed area fed by the septal branch used for ethanol injection. No remote effects were observed. TASH leads to a sharply delineated septal reduction of perfusion and metabolism with consecutive reduction of septal thickness, a sustained elimination of the outflow tract obstruction, and a substantial clinical improvement without impairment of global ventricular function.


Modeling nikkomycin Z dosing and pharmacology in murine pulmonary coccidioidomycosis preparatory to phase 2 clinical trials.
Nikkomycin Z (NikZ) is a chitin synthase inhibitor with activity against Coccidioides species that is being developed as a first-in-class orphan product for treatment of coccidioidomycosis. It has previously been shown to reduce lethal respiratory infections in mice to undetectable levels when treatment is begun 48 hours after infection. The studies described here focus on bracketing NikZ doses for phase 2 and 3 clinical trials, using an established mouse respiratory infection as a model and starting treatment 120 hours after infection. A dose of 80 mg/kg/day, divided into 2 doses, nearly eradicated infection, and larger doses did not improve fungal clearance. Increasing the duration of treatment from 1 week to 3 weeks resulted in a greater percentage of culture-negative mice. Comparative data show that plasma levels of NikZ that nearly eradicate Coccidioides in mice are achievable in patients and provide a plausibly effective dose range for initial phase 2 clinical studies.


Sensory denervation of the kidney attenuates renovascular hypertension in the rat.
To determine the role of the renal afferent nerves in the pathogenesis of one-kidney, one-clip renovascular hypertension, the renal afferent nerves were selectively lesioned by dorsal rhizotomy, a procedure that eliminates renal sensory input to the spinal cord but does not directly damage the sympathomotor innervation of the kidney. One week after denervation, the proximal left renal artery was clipped in denervated and sham control rats. Blood pressure of the sham group rose progressively over the next 5 wk, to 185 mmHg (systolic). In contrast, blood pressure of the denervated rats leveled off in the borderline hypertensive range, a level significantly lower than that of the sham group but significantly higher than that of non-clipped rats. In two further experiments the specificity of this effect was demonstrated. Lesion of the dorsal root nerves on the side of nephrectomy did not significantly lower blood pressure of non-clipped rats, and contralateral dorsal rhizotomy did not lower the blood pressure of clipped rats. These results demonstrate that the renal afferent nerves significantly contribute to one-kidney, one-clip renovascular hypertension in the rat.


Integrated Liver and Plasma Proteomics in Obese Mice Reveals Complex Metabolic Regulation.
Obesity leads to the development of nonalcoholic fatty liver disease (NAFLD) and associated alterations to the plasma proteome. To elucidate the underlying changes associated with obesity, we performed liquid chromatography-tandem mass spectrometry in the liver and plasma of obese leptin-deficient ob/ob mice and integrated these data with publicly available transcriptomic and proteomic datasets of obesity and metabolic diseases in preclinical and clinical cohorts. We quantified 7173 and 555 proteins in the liver and plasma proteomes, respectively. The abundance of proteins related to fatty acid metabolism were increased, alongside peroxisomal proliferation in ob/ob liver. Putatively secreted proteins and the secretory machinery were also dysregulated in the liver, which was mirrored by a substantial alteration of the plasma proteome. Greater than 50% of the plasma proteins were differentially regulated, including NAFLD biomarkers, lipoproteins, the 20S proteasome, and the complement and coagulation cascades of the immune system. Integration of the liver and plasma proteomes identified proteins that were concomitantly regulated in the liver and plasma in obesity, suggesting that the systemic abundance of these plasma proteins is regulated by secretion from the liver. Many of these proteins are systemically regulated during type 2 diabetes and/or NAFLD in humans, indicating the clinical importance of liver-plasma cross talk and the relevance of our investigations in ob/ob mice. Together, these analyses yield a comprehensive insight into obesity and provide an extensive resource for obesity research in a prevailing model organism.


Intrauterine negative-pressure therapy (IU-NPT) to treat peritonitis after caesarean section.
We describe the first application of intrauterine negative-pressure therapy (IU-NPT) for an early rupture of a uterine suture after a third caesarean section with consecutive peritonitis and sepsis. Because all four quadrants were affected by peritonitis, a laparotomy was performed on the 15th day after caesarean section. Abdominal negative-pressure wound therapy (A-NPWT) of the open abdomen was initiated. During the planned relaparotomy, a suture defect of the anterior uterine wall was identified and sutured. In the second relaparotomy, the suture appeared once more insufficient. For subsequent IU-NPT, we used an open-pore film drainage (OFD) consisting of a drainage tube wrapped in the double-layered film. The OFD was inserted into the uterine cavity via the uterine defect and IU-NPT was established together with A-NPT. With the next relaparotomy, local inflammation and peritonitis had been resolved completely. IU-NPT was continued transvaginally, the uterine defect was sutured, and the abdomen was closed. Vaginal IU-NPT was also discontinued after another eight days. By using IU-NPT, local infection control of the septic focus was achieved. The infectious uterine secretions were completely evacuated and no longer discharged into the abdominal cavity. As a result of the applied suction, the uterine cavity collapsed around the inlaid OFD. The total duration of IU-NPT was 11 days. The uterine defect was completely closed, and a hysterectomy was avoided. The patient was discharged four days after the end of IU-NPT. IU-NPT follows the same principles as those described for endoscopic negative-pressure wound therapy of the gastrointestinal tract.


Vocal plasticity in a reptile.
Sophisticated vocal communication systems of birds and mammals, including human speech, are characterized by a high degree of plasticity in which signals are individually adjusted in response to changes in the environment. Here, we present, to our knowledge, the first evidence for vocal plasticity in a reptile. Like birds and mammals, tokay geckos (Gekko gecko) increased the duration of brief call notes in the presence of broadcast noise compared to quiet conditions, a behaviour that facilitates signal detection by receivers. By contrast, they did not adjust the amplitudes of their call syllables in noise (the Lombard effect), which is in line with the hypothesis that the Lombard effect has evolved independently in birds and mammals. However, the geckos used a different strategy to increase signal-to-noise ratios: instead of increasing the amplitude of a given call type when exposed to noise, the subjects produced more high-amplitude syllable types from their repertoire. Our findings demonstrate that reptile vocalizations are much more flexible than previously thought, including elaborate vocal plasticity that is also important for the complex signalling systems of birds and mammals. We suggest that signal detection constraints are one of the major forces driving the evolution of animal communication systems across different taxa.


Impact of lifetime traumatic experiences on suicidality and likelihood of conversion in a cohort of individuals at clinical high-risk for psychosis.
Recent research suggests that trauma history (TH) is a strong socio-environmental risk factor for the development of psychosis. While reported rates of childhood trauma are higher among individuals at clinical high-risk (CHR) for psychosis than in the general population, little research has explored the effects of trauma upon the severity of attenuated positive symptoms. We aimed to explore the specific relationships between TH and baseline symptom severity; likelihood of conversion to full-blown psychosis; suicidal ideation (SI); and suicidal behavior (SB) in a cohort of 200 help-seeking CHR individuals. Participants were evaluated every three months for up to two years using the Structured Interview for Psychosis-Risk Syndromes (SIPS). More trauma history was reported by females and Hispanic/Latino participants, while age and race did not significantly distinguish those with and without TH. Individuals with TH reported higher rates of SI and SB than those without. While TH was positively associated with several SIPS subscales, including Unusual Thought Content, Perceptual Abnormalities/Hallucinations, Bizarre Thinking, Sleep Disturbances, and Dysphoric Mood, and negatively associated with Expressed Emotion, results indicated that TH was not significantly related to conversion to psychosis. Moreover, baseline SI was unrelated to conversion and baseline DSM diagnosis, with the exception of Post-Traumatic Stress Disorder (PTSD). These results suggest that traumatic experiences may significantly impact the severity of attenuated positive symptoms and suicidality in the CHR state, providing new windows for further research and potential intervention.


Synthesis of message and genome RNAs in vitro by Sendai virus-infected cell nucleocapsids.
Purified Sendai virus nucleocapsids isolated from infected cells were used to programme a transcription system in vitro to study virus-specific RNA synthesis. The RNA products were analysed for size by centrifugation before and after denaturation with formamide or glyoxal. The polarity of the products [message (+) or genome (-) strands] was analysed by RNA-RNA hybridization. The non-denatured RNA products sedimented in three groups: 7S to 22S single-stranded RNA transcripts and two partially ribonuclease-resistant complexes. One complex, representing 12% of the total product, sedimented at 26S to 36S. After denaturing the 26S to 36S complex to single-stranded molecules, about half of the RNAs sedimented at 25S to 54S and about half at 6S to 24S. The second complex, representing about 13% of the total RNA product, sedimented at 42S to 52S. After denaturing, about 10% of the single-stranded RNAs sedimented at 38S to 52S and about 90% sedimented at 6S to 19S. In hybridization studies, single-stranded RNAs that sedimented at less than 19S were predominantly of message sense (+ strand), whereas RNAs that sedimented at 25S to 54S were a mixture of genome and anti-genome type. These results show that transcription and replication activities in vitro were associated with Sendai virus nucleocapsids obtained from infected cells and that some of the reaction products approached genome size.


Speech and language therapy interventions for children with primary speech and language delay or disorder.
It is thought that approximately 6% of children have speech and language difficulties of which the majority will not have any other significant developmental difficulties. Whilst most children's difficulties resolve, children whose difficulties persist into primary school may have long-term problems concerning literacy, socialisation, behaviour and school attainment. To examine the effectiveness of speech and language interventions for children with primary speech and language delay/disorder. The following databases were searched: The Cochrane Controlled Trials Register (Cochrane Library, CENTRAL: 2002/3), CINAHL (1982 - July 2002), EMBASE (1980 - Sept Week 4 2002), ERIC (1965 - 2002), MEDLINE (1966 - Sept Week 3 2002), PsycINFO (1872 - 2002/10 Week 2), The National Research Register (2002/3). In addition to this references were taken from reviews of the literature and reference lists from articles. The review considered randomised controlled trials of speech and language therapy interventions for children or adolescents with primary speech and language delay/disorder. Titles and abstracts were identified and assessed for relevance, before the full text version was obtained of all potentially relevant articles. The data were categorised depending on the nature of the control group and considered in terms of the effects of intervention on expressive and receptive phonology, syntax and vocabulary. The outcomes used in the analysis were dependent on the focus of the study with only the primary effects of therapy being considered in this review. The results of twenty-five studies were used in the meta-analysis. The results suggest that speech and language therapy is effective for children with phonological (SMD=0.44, 95%CI: 0.01,0.86) or vocabulary difficulties (SMD=0.89, 95%CI: 0.21,1.56), but that there is less evidence that interventions are effective for children with receptive difficulties (SMD=-0.04, 95%CI: -0.64,0.56). Mixed findings were found concerning the effectiveness of expressive syntax interventions (n=233; SMD=1.02, 95%CI: 0.04-2.01). No significant differences were shown between clinician administered intervention and intervention implemented by trained parents, and studies did not show a difference between the effects of group and individual interventions (SMD=0.01, 95%CI: -0.26,1.17). The use of normal language peers in therapy was shown to have a positive effect on therapy outcome (SMD=2.29, 95%CI: 1.11,3.48). The review shows that overall there is a positive effect of speech and language therapy interventions for children with expressive phonological and expressive vocabulary difficulties. The evidence for expressive syntax difficulties is more mixed, and there is a need for further research to investigate intervention for receptive language difficulties. There is a large degree of heterogeneity in the results, and the sources of this need to be investigated.


Prevalence of outpatient cancer treatment in the United States: estimates from the Medical Panel Expenditures Survey (MEPS).
Little is known regarding the prevalence of outpatient cancer treatment in the U.S. We analyzed nationally-representative data from the 2000-2004 Medical Expenditure Panel Survey to estimate the number of U.S. cancer patients receiving outpatient chemotherapy and/or radiation therapy annually. Each year, over 1.1 million individuals are estimated to receive chemotherapy or radiation therapy for cancer. Cancer patients younger than 65 receiving treatment who were uninsured were less likely to receive chemotherapy or combined chemotherapy/radiation therapy than were those with public or private insurance. These estimates may be useful for understanding the burden of cancer care and development of programs for cancer survivors.


Two metals are better than one in the gold catalyzed oxidative heteroarylation of alkenes.
We present a detailed study of the mechanism for oxidative heteroarylation, based on DFT calculations and experimental observations. We propose binuclear Au(II)-Au(II) complexes to be key intermediates in the mechanism for gold catalyzed oxidative heteroarylation. The reaction is thought to proceed via a gold redox cycle involving initial oxidation of Au(I) to binuclear Au(II)-Au(II) complexes by Selectfluor, followed by heteroauration and reductive elimination. While it is tempting to invoke a transmetalation/reductive elimination mechanism similar to that proposed for other transition metal complexes, experimental and DFT studies suggest that the key C-C bond forming reaction occurs via a bimolecular reductive elimination process (devoid of transmetalation). In addition, the stereochemistry of the elimination step was determined experimentally to proceed with complete retention. Ligand and halide effects played an important role in the development and optimization of the catalyst; our data provides an explanation for the ligand effects observed experimentally, useful for future catalyst development. Cyclic voltammetry data is presented that supports redox synergy of the Au···Au aurophilic interaction. The monometallic reductive elimination from mononuclear Au(III) complexes is also studied from which we can predict a ~15 kcal/mol advantage for bimetallic reductive elimination.


Arthroscopic findings of coexisting lesions with greater tuberosity fractures.
Proximal humerus fractures comprise approximately 5% of all fractures, with isolated greater tuberosity fractures accounting for approximately 20% of proximal humerus fractures. Although performing shoulder arthroscopy in situations including a fracture is technically demanding, it allows surgeons the opportunity to identify and treat other coexisting lesions that could have otherwise been missed. The incidence of these pathologies in combination with greater tuberosity fractures has not been established. This study aimed to identify the various types of pathologies that may coexist with greater tuberosity fractures but not be detected before fixation. Displaced 2-part greater tuberosity fractures were treated arthroscopically in the authors' department. All patients initially underwent diagnostic arthroscopy during which other coexisting pathologies were detected and assessed, including rotator cuff tears, labral tears (Bankart or superior labral anterior posterior lesions), or long head of the biceps pathologies. Twenty-four patients underwent arthroscopic (n=10) or arthroscopic-assisted (n=14) greater tuberosity reduction and fixation. Thirteen (54.2%) fragments were fully displaced. Four (16.7%) patients had fracture dislocation of the glenohumeral joint. The concomitant soft-tissue pathologies were identified and treated arthroscopically in 22 (92%) patients. Arthroscopic evaluation before greater tuberosity fracture fixation revealed a high percentage of concomitant soft tissue pathologies. These pathologies may be overlooked otherwise, but they are easily detected arthroscopically, enabling their treatment during the same procedure.


Excluded facility financial status and options for payment system modification.
Psychiatric, rehabilitation, long-term care, and children's facilities have remained under the reimbursement system established under the Tax Equity and Fiscal Responsibility Act (TEFRA) of 1982 (Public Law 97-248). The number of TEFRA facilities and discharges has been increasing while their average profit rates have been steadily declining. Modifying TEFRA would require either rebasing the target amount or adjusting cost sharing for facilities exceeding their cost target. Based on our simulations of alternative payment systems, we recommend rebasing facilities' target amounts using a 50/50 blend of own costs and national average costs. Cost sharing above the target amount could be increased to include more government sharing of losses.


The emergence of gender differences in physical aggression in the context of conflict between young peers.
It is well known that a gender difference in physical aggression emerges by the preschool years. We tested the hypothesis that the gender difference is partly due to changing tactics in peer interaction. Observations of girls' and boys' social initiatives and reactions to opportunities for conflict were made, using the Peer Interaction Coding System (PICS) in four independent samples of children between 9 and 36 months of age, which were aggregated to form a summary data set (N= 323), divided into two age bands (below or above 24 months of age). Linear mixed-model analyses revealed significant age by gender interactions in the use of bodily force in response to peers' initiatives and in the tendency to use bodily force at later stages of conflicts with peers. The gender difference in use of force was not explained by differences in the use of verbal tactics. These cross-sectional findings suggest that girls are initially more likely than boys to use reactive aggression, but then desist, whereas boys increase their use of force to defend their territory and possessions. The difference between older and younger girls likely reflects girls' abilities to regulate their behaviour in response to social challenges and the fact that girls are explicitly socialized to yield to peers' demands.


Associations between inflammatory and immune response genes and adverse respiratory outcomes following exposure to outdoor air pollution: a HuGE systematic review.
Variants of inflammatory and immune response genes have been associated with adverse respiratory outcomes following exposure to air pollution. However, the genes involved and their associations are not well characterized, and there has been no systematic review. Thus, we conducted a review following the guidelines of the Human Genome Epidemiology Network. Six observational studies and 2 intervention studies with 14,903 participants were included (2001-2010). Six studies showed at least 1 significant gene-pollutant interaction. Meta-analysis was not possible due to variations in genes, pollutants, exposure estimates, and reported outcomes. The most commonly studied genes were tumor necrosis factor α (TNFA) (n = 6) and toll-like receptor 4 (TLR4) (n = 3). TNFA -308G>A modified the action of ozone and nitrogen dioxide on lung function, asthma risk, and symptoms; however, the direction of association varied between studies. The TLR4 single-nucleotide polymorphisms rs1927911, rs10759931, and rs6478317 modified the association of particulate matter and nitrogen dioxide with asthma. The transforming growth factor β1 (TGFB1) polymorphism -509C>T also modified the association of pollutants with asthma. This review indicates that genes controlling innate immune recognition of foreign material (TLR4) and the subsequent inflammatory response (TGFB1, TLR4) modify the associations of exposure to air pollution with respiratory function. The associations observed have biological plausibility; however, larger studies with improved reporting are needed to confirm these findings.


Prevalence and Patterns of Multimorbidity among Human Immunodeficiency Virus Positive People in Odisha, India: An Exploratory Study.
Improved and efficient Anti-Retroviral Therapy (ART) has turned HIV positive into a chronic disorder. The increase in life expectancy and quality of life comes with an increased risk of multimorbidity from the traditional risk factors. To explore the prevalence of multimorbidity among People Living with HIV (PLHIV). An exploratory cross-sectional study was carried out in an ART center of Odisha, India, from November 2013 to December 2013. A systematic random sampling was done to include 197 adult HIV positive patients receiving ART at least for one year or more. A structured questionnaire was used to collect data on socio-demographic profile, ART status, multimorbidity and opportunistic infections. The prevalence of multimorbidity was 47.7% (95% CI 41% to 55%). Among them, the most common chronic conditions were pulmonary tuberculosis (28.4%), gastro-intestinal conditions (7.1%), neural disorders (5.1%) and hypertension (2.5%). In total, 48.7% had opportunistic infections; with 36% bacterial, 25% fungal and 21% viral infections. The prevalence of multimorbidity was high among HIV positive patients. Further research should explore the epidemiology of multimorbidity among HIV positive patients to formulate proper clinical practice guidelines to enhance long-term outcomes of PLHIV in the current treatment era.


[Validation of a questionnaire for the diagnosis of headache in an outpatient clinic at a university hospital].
Headache is a common complaint with a high prevalence in ambulatory settings. The physical and neurological examinations are frequently normal. The use of questionnaires as a screening methods for patients with primary headache could facilitate the diagnosis in non-specialized medical centers. In the present study, we used a questionnaire, based on the IHS criteria and modified by the authors, applied to 204 patients from the outpatient clinic of the Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo. Half of the patients were submitted to a clinical interview. We compared the results of the questionnaire with the results of the medical interview (gold standard). Most of the headaches we studied were primary headaches (89.6%). The questionnaire revealed a sensitivity of 90.2% and specificity of 57.9% for migraine detection with a chance corrected agreement (kappa) of 0.47 and a positive predictive (PPV) value of 65.7% and a negative predictive value (NPV) de 86.8%. The sensitivity for tension-type headache detection was 60.8% and the specificity 87.1% with a kappa value of 0.49 and a PPV of 77.8% and a NPV of 75.9%. We conclude that this questionnaire can be used as a screening method for diagnosing headache and that it can be applied by non-medical personnel. This questionnaire could also be used for population studies.


Prevalence and severity of menopause symptoms and associated factors across menopause status in Korean women.
The present study investigated the prevalence and severity of menopause symptoms experienced by Korean women aged 44 to 56 years and their associated factors. A cross-sectional study was performed on 2,201 women aged 44 to 56 years in health checkup centers between November 2012 and March 2013. The 29-item Menopause-Specific Quality of Life Questionnaire was used to assess vasomotor, psychosocial, physical, and sexual symptoms related to menopause. The guidelines for the classification of reproductive aging stages proposed at the Stages of Reproductive Aging Workshop were used. Multivariable linear regression analyses were performed to identify factors associated with severity of menopause symptoms. Among participants, 42.6% were premenopausal, 36.7% were perimenopausal, and 20.7% were postmenopausal. Although physical symptoms were the most severe menopause symptoms experienced by premenopausal and perimenopausal women, postmenopausal women reported sexual symptoms as the most bothersome. The mean scores for each domain increased from the premenopausal period through the postmenopausal period (P for trend < 0.001). The regression model revealed that age (for vasomotor and sexual symptoms) and obesity (for vasomotor and physical symptoms) were significantly associated with severity of menopause symptoms (P < 0.05). Physically active women had fewer severe physical symptoms related to menopause than inactive women. Postmenopausal women experience the most severe symptoms. Obesity and physical activity are the main modifiable factors associated with symptom severity. Further studies are needed to examine the effects of physical activity promotion and weight control interventions on preventing menopause symptoms in Korean women.


Magnetic field effects in nematic and smectic liquid crystals probed by time resolved observation of orientation relaxation of the spin probe.
The kinetics of reorientation of the liquid crystal HOPOOB in nematic and smectic phases in the magnetic field was studied by the spin probe technique. The time evolution of the EPR spectrum shape in the course of reorientation in the nematic phase indicates that the mechanism of alignment comprises redistribution between domains of different orientations rather than the turn of the orientation distribution function. A microscopic model of domain structure rearrangement is provided to qualitatively describe the experimental data. In the smectic C phase, partial reorientation is observed upon the action of the magnetic field. A possible mechanism of this process is the reorientation of tilt direction within the smectic layers. It is shown that this process is constrained, and the value of the corresponding elastic constant is estimated.


An MYC-Driven Vicious Circuit Is a Targetable Achilles' Heel in Lymphoma.
In this issue of Blood Cancer Discovery, Nakanishi et al. uncover a critical role for the elevated activity of the translation initiation factor eIF5A in the malignant growth of MYC-driven lymphoma. eIF5A is posttranslationally modified by hypusination through MYC oncoprotein-mediated hyperactivation of the polyamine-hypusine circuit, which may represent a promising therapeutic target because an enzyme of this circuit that is required for hypusinating eIF5A proved to be essential for lymphoma development. See related article by Nakanishi et al., p. 294 (4).


The clinical characteristics and correlates of lithium toxicity in a tertiary referral centre.
Lithium is a medication indicated for the treatment of bipolar disorder and treatment-resistant depression, with a narrow therapeutic index. Overdose, either acute or chronic can result in neurological symptoms, requiring dialysis and admission to intensive care in some cases. Lithium toxicity is avoidable with careful monitoring. However, we have noted several recent cases of lithium toxicity in our local service and thus sought to investigate this issue in a more systematic manner. We aimed to quantify the incidence of lithium toxicity in our local population over a single year and identify the patients most at risk. We also aimed to generate clinical recommendations on the prevention of lithium toxicity to improve patient safety. We identified the incidence of lithium toxicity in our local population, by searching the hospital pathology database for patients with serum lithium levels greater than 1.0 mmol/L. We examined the available clinical notes for these patients. We identified 74 serum lithium readings above 1.0 mmol/L measured in 44 individual patients. The highest recorded level was 3.2 mmol/L. Of these, 11 patients were aged 65 years or older. Hospital admission was required in 14 cases. There were missing data of note: 29.5% had no renal function/eGFR measurement at time of toxicity and 52.3% without a baseline eGFR. Lithium toxicity is common in our population. Given the narrow therapeutic index, this demonstrates the need for careful monitoring and prescribing, especially patients aged 65 and over.


Modelling polycystic liver disease progression using age-adjusted liver volumes and targeted mutational analysis.
Polycystic liver disease (PLD) manifests as numerous fluid-filled cysts scattered throughout the liver parenchyma. PLD most commonly develops in females, either as an extra-renal manifestation of autosomal-dominant polycystic kidney disease (ADPKD) or as isolated autosomal-dominant polycystic liver disease (ADPLD). Despite known genetic causes, clinical variability challenges patient counselling and timely risk prediction is hampered by a lack of genotype-phenotype correlations and prognostic imaging classifications. We performed targeted next-generation sequencing and multiplex ligation-dependent probe amplification to identify the underlying genetic defect in a cohort of 80 deeply characterized patients with PLD. Identified genotypes were correlated with total liver and kidney volume (assessed by CT or MRI), organ function, co-morbidities, and clinical endpoints. Monoallelic diagnostic variants were identified in 60 (75%) patients, 38 (48%) of which pertained to ADPKD-gene variants (PKD1, PKD2, GANAB) and 22 (27%) to ADPLD-gene variants (PRKCSH, SEC63). Disease severity defined by age at waitlisting for liver transplantation and first PLD-related hospitalization was significantly more pronounced in mutation carriers compared to patients without genetic diagnoses. While current imaging classifications proved unable to differentiate between severe and moderate courses, grouping by estimated age-adjusted total liver volume progression yielded significant risk discrimination. This study underlines the predictive value of providing a molecular diagnosis for patients with PLD. In addition, we propose a novel risk-classification model based on age- and height-adjusted total liver volume that could improve individual prognostication and personalized clinical management. Polycystic liver disease (PLD) is a highly variable condition that can be asymptomatic or severe. However, it is currently difficult to predict clinical outcomes such as hospitalization, symptom burden, and need for transplantation in individual patients. In the current study, we aimed to investigate the clinical value of genetic confirmation and an age-adjusted total liver volume classification for individual disease prediction. While genetic confirmation generally pointed to more severe disease, estimated age-adjusted increases in liver volume could be useful for predicting clinical outcomes.


A twin study of early cannabis use and subsequent use and abuse/dependence of other illicit drugs.
Cannabis use is strongly associated with the use and abuse/dependence of other illicit drugs. Gateway and common liabilities models have been employed to explain this relationship. We sought to examine this association using a combination of the discordant twin design and modeling methods. We assess the relationship between early cannabis use and the subsequent use and abuse/ dependence of other illicit drugs in a population-based sample of male and female twin pairs using four analyses: (i) analysis of the association between early cannabis use and other illicit drug use and abuse/dependence in the entire sample of twins, (ii) assessment of the influence of early cannabis use in twin 1 on twin 2's use or abuse/dependence of other illicit drugs, (iii) use of twin pairs discordant for early cannabis use in a discordant twin design and (iv) a model-fitting procedure. We found: (i) a strong association between early cannabis use and use and abuse/dependence of other illicit drugs in the sample, (ii) twin 1's early cannabis use is significantly associated with the twin 2's other illicit drug use, (iii) the role of correlated genetic factors with some evidence for a causal influence, and (iv) the correlated liabilities model fits the data well. Early cannabis use is strongly associated with other illicit drug use and abuse/dependence. The relationship arises largely due to correlated genetic and environmental influences with persisting evidence for some causal influences.


CP2 binding to the promoter is essential for the enhanced transcription of globin genes in erythroid cells.
We have previously reported that the reduced level of CP2 suppresses the mouse alpha- and beta-globin gene expression and hemoglobin synthesis during terminal differentiation of mouse erythroleukemia (MEL) cells in vitro [Chae et al. (1999)]. As an extension of this study, we demonstrated that human alpha-, epsilon-, and gamma- globin genes were also suppressed by the reduced expression of CP2 in K562 cells. To address how much CP2 contributes in the regulation of globin gene expression, we measured transcriptional activities of the wild type alpha-globin promoter and its various factor-binding sites mutants in erythroid and nonerythroid cells. Interestingly, CP2 site dependent transcriptional activation occurred in an erythroid-cell specific manner, even though CP2 is ubiquitously expressed. In addition, CP2 site mutation within the alpha-promoter severely suppressed promoter activity in differentiated, but not in undifferentiated MEL cells, suggesting that the CP2 binding site is needed for the enhanced transcription of globin genes during erythroid differentiation. When the human beta-globin locus control region was linked to the alpha-promoter, suppression was more severe in the CP2 site mutant in differentiated MEL cells. Overall data indicate that CP2 is a major factor in the regulation of globin expression in human and mouse erythroid cells, and CP2 binding to the globin gene promoter is essential for the enhanced transcription of globin genes in erythroid differentiation.


Integrating Immune Therapies for the Treatment of Multiple Myeloma.
Patients with relapsed or refractory multiple myeloma (RRMM) that is refractory to a proteasome inhibitor, an immunomodulatory drug (IMiD), and an anti-CD38 antibody (triple-class refractory MM) have poor outcomes. Recently, 2 classes of T-cell engaging therapies-CAR T-cell therapy and bispecific T-cell engaging antibodies (BsAbs)-have resulted in unprecedented response rates and survival outcomes in these heavily pretreated patients. The most common targets are BCMA and GPRC5D, with other targets in development. The main classes of adverse effects include cytokine release syndrome, neurotoxicity, infections, and cytopenias, as well as adverse effects unique to specific products. As of September 2023, 2 BCMA-targeting CAR-T cell products, 2 BCMA-targeting BsAbs, and 1 GPRC5D-targeting BsAb, are FDA-approved for standard-of-care use in patients with RRMM who received at least 4 prior lines of therapy, including prior treatment with a proteasome inhibitor, an IMiD, and an anti-CD38 antibody. Earlier-line use is under investigation and has shown promising results. Several other investigational CAR-T constructs and bispecific antibodies are in clinical development. As these therapies become more widely used, including in earlier-line setting, efforts to understand optimal sequencing and mitigate toxicities remain critical.


The effect of ultrasound on bone dimensional changes following extraction: a pilot study.
Because of bone resorption following tooth extraction, preservation of adequate bony dimension is performed often for subsequent treatment with dental implants. This study evaluated a novel, non-invasive treatment using ultrasound to accelerate healing following extraction to minimize alveolar bone loss. The objective of this study was to evaluate the effect of ultrasound on the dimensional healing changes of alveolar bone following tooth extraction using cone-beam volumetric tomography (CBVT). This randomized, split-mouth trial involved 12 subjects requiring extraction of two contralateral erupted permanent teeth. Baseline CBVT scans were captured 7 to 10 days following extraction, after which time ultrasound therapy commenced in test sites. Ultrasound therapy was delivered for 20 minutes using a piezoelectric transducer for 10 sessions over the subsequent 4 weeks. Follow-up radiographic scans were obtained at 4 weeks and 3 months postextraction. Analyses of variance and covariance were performed to assess dimensional changes over the 3-month healing period. Analysis of dimensional changes in all measures of vertical height and horizontal width demonstrated no statistically significant differences between the ultrasound and control groups from baseline to 3 months postextraction. Evaluation of correlations between dimensional changes demonstrated a moderately strong correlation (r = 0.67; P = 0.023) in the ultrasound group between the change in buccal vertical height and the baseline crestal ridge width. Analysis of the change in buccal vertical height relative to baseline crestal width demonstrated a statistically significant benefit to ultrasound compared to control (P = 0.016). This benefit was more pronounced in wider sockets compared to narrow sockets. In this pilot study, there was no significant benefit to ultrasound in absolute bony dimensional changes following tooth extraction. There was a significant interaction between the treatment rendered (ultrasound versus control) and the change in buccal ridge height relative to baseline ridge width at the crest and 3 mm apical to the crest. This benefit was apparent in wide sockets compared to narrow sockets; however, the clinical importance of these relative dimensional changes in the ultrasound group are difficult to determine given the inclusion of all tooth types in a pilot study with a small sample size.


Tumor necrosis factor-alpha signals to the IFN-gamma receptor complex to increase Stat1alpha activation.
We describe a novel mechanism of signaling interaction through which tumor necrosis factor-alpha (TNF-alpha) treatment augments interferon-gamma (IFN-gamma)-induced Stat1alpha DNA-binding complexes and transcriptional activation of a Stat-binding element. In TNF-alpha-treated cells, IFN-gamma-induced phosphorylation of Jak2 kinase is increased, Jak2 kinase activity is enhanced, and genetic studies indicate that TNF-alpha requires Jak2 kinase activity to enhance Stat1alpha tyrosine phosphorylation. Increased Jak2 and Stat1alpha phosphorylation are observed within minutes of coexposure to TNF-alpha/IFN-gamma, suggesting a direct signaling interaction. IFN-gamma receptor chain 1 (IFNGR-1) tyrosine phosphorylation is markedly enhanced in cells treated with TNF-alpha/IFN-gamma without alteration in receptor levels. Thus, there exists a direct signaling interaction between TNF-alpha and IFN-gamma, independent of cooperating enhancer elements, that may be relevant for cytokine action during immune-mediated host defense and inflammatory processes.


Health care providers' experience reporting child abuse in the primary care setting. Pediatric Practice Research Group.
To describe (1) primary care providers' experiences identifying and reporting suspected child abuse to child protective services (CPS) and (2) variables affecting providers' reporting behavior. Health care providers (76 physicians, 8 nurse practitioners, and 1 physician assistant) in a regional practice-based network completed written surveys that collected information about the demographic characteristics of each provider and practice; the provider's career experience with child abuse; and the provider's previous year's experience identifying and reporting suspected child abuse, including experience with CPS. All providers (N = 85) in 17 participating practices completed the survey. In the preceding 1 year, 48 respondents (56%) indicated that they had treated a child they suspected was abused, for an estimated total of 152 abused children. Seven (8%) of 85 providers did not report a total of 7 children with suspected abuse (5% of all suspected cases). A majority of providers (63%; n = 29) believed that children who were reported had not benefited from CPS intervention, and 21 (49%) indicated that their experience with CPS made them less willing to report future cases of suspected abuse. Providers who had some formal education in child abuse after residency were 10 times more likely to report all abuse than were providers who had none. Primary care providers report most, but not all, cases of suspected child abuse that they identify. Past negative experience with CPS and perceived lack of benefit to the child were common reasons given by providers for not reporting. Education increases the probability that providers will report suspected abuse.


Underestimation of pelvic osteolysis: the value of the iliac oblique radiograph.
Periacetabular osteolysis is recognized as a complication of cementless total hip arthroplasty. Routine follow-up radiographs are recommended by most joint reconstructive surgeons to detect osteolytic lesions that may remain asymptomatic. These studies usually consist of an anteroposterior (AP) pelvic and lateral radiograph of the hip. Three male cadaver pelves were used to evaluate whether a routine AP pelvic radiograph was adequate to show certain osteolytic lesions. This study involved creation of progressively enlarging defects simulating an osteolytic lesion of the posterior wall. The extent of involvement of the lesion as shown by an AP pelvic radiograph and iliac oblique views was then compared. By the time the posterior column lesion could be estimated to the same extent on both the AP pelvic and iliac oblique radiographs, the lesion had become uncontained in the posterior medial direction. The evaluation of pelvic osteolysis with biplanar imaging provides a 3-dimensional understanding of the lesion. This study shows the inadequacy of a single AP pelvic radiograph in showing and localizing periprosthetic osteolytic lesions in the posterior column.


[Behavior of the cardiovascular system in outer space].
One of the important factors in outer space is the absence of gravity (OG). During longterm missions, this factor is responsible for the larger number of anatomical and physiological changes that astronauts experience. The cardiovascular system undergoes these changes with severe intensity, which is part of an adaptation process to the new environmental conditions. The modifications observed in both the anatomy of the cardiovascular system and its hemodynamics occur in two phases. The first phase begins when the astronauts enter into Earth orbit or in interplanetary trajectory and extends until the second or fourth day of the mission. It is characterized by an important shifting of fluids from the lower extremities to the cephalic regions which produces an increase of the venous return and the preload, the heart rate is increased, the blood volume in the thorax is also increased, the cardiac chambers become dilated, and by reflex action, the antidiuretic hormone diminishes, diuresis increases and leads to a virtual state of dehydration. Clinically, the first stage is manifested by headache, dizziness, space disorientation, nausea, anorexia, projectile vomiting, sweating and pallor. This constalation of data is known as "The Space Adaptation Syndrome". The second phase begins at the end of the first phase and finishes toward the fortieth or fiftieth day of the mission.(ABSTRACT TRUNCATED AT 250 WORDS)


Membrane raft organization is more sensitive to disruption by (n-3) PUFA than nonraft organization in EL4 and B cells.
Model membrane and cellular detergent extraction studies show (n-3) PUFA predominately incorporate into nonrafts; thus, we hypothesized (n-3) PUFA could disrupt nonraft organization. The first objective of this study was to determine whether (n-3) PUFA disrupted nonrafts of EL4 cells, an extension of our previous work in which we discovered an (n-3) PUFA diminished raft clustering. EPA or DHA treatment of EL4 cells increased plasma membrane accumulation of the nonraft probe 1,1'-dilinoleyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate by ~50-70% relative to a BSA control. Förster resonance energy transfer imaging showed EPA and DHA also disrupted EL4 nanometer scale nonraft organization by increasing the distance between nonraft molecules by ~25% compared with BSA. However, changes in nonrafts were due to an increase in cell size; under conditions where EPA or DHA did not increase cell size, nonraft organization was unaffected. We next translated findings on EL4 cells by testing if (n-3) PUFA administered to mice disrupted nonrafts and rafts. Imaging of B cells isolated from mice fed low- or high-fat (HF) (n-3) PUFA diets showed no change in nonraft organization compared with a control diet (CD). However, confocal microscopy revealed the HF (n-3) PUFA diet disrupted lipid raft clustering and size by ~40% relative to CD. Taken together, our data from 2 different model systems suggest (n-3) PUFA have limited effects on nonrafts. The ex vivo data, which confirm previous studies with EL4 cells, provide evidence that (n-3) PUFA consumed through the diet disrupt B cell lipid raft clustering.


Online programs improve relationship functioning for distressed low-income couples: Results from a nationwide randomized controlled trial.
Although low-income couples experience greater relationship challenges, they have limited access to effective relationship interventions. Furthermore, most previous efforts to improve low-income couples' relationships have yielded very small effects (Hawkins & Erickson, 2015). In an effort to overcome these limitations, this study investigated the effectiveness of 2 web-based interventions for low-income couples. In total, 742 low-income couples (N = 1,484 individuals; mean [M] age = 33; 55% White, non-Hispanic; 52% married; median [Mdn] annual household income = $27,000) were recruited nationally and randomized to the OurRelationship program, the ePREP program, or a waitlist control group. Couples were repeatedly assessed for 6 months using self-report measures of relationship satisfaction, communication conflict, intimate partner violence, emotional support, and breakup potential. Relationship status was assessed at 6-month follow-up. Compared to the control group, intervention couples experienced significantly greater improvements in all 5 domains of relationship functioning (Mdn |d| = 0.46) by the end of the program; these effects were maintained in the 4 months after treatment. However, neither program significantly reduced the frequency of breakups by the 6-month follow-up. Differences between couples in the two interventions were minimal (Mdn Cohen's |d| = 0.11); however, couples in the OurRelationship program experienced significantly greater decreases in conflict (d = 0.24). The results indicate that brief, web-based interventions can serve a central role in delivering effective services to low-income couples. Additionally, the general equivalence of the two interventions indicates that both communication-focused and problem-focused interventions can be successful in improving the relationship functioning of low-income couples. (PsycINFO Database Record (c) 2020 APA, all rights reserved).


Association between prepregnancy obesity and metabolic risk in Chilean premenopausal women 10 y postpartum.
One of every four pregnant women in Chile is obese. Gestational obesity is associated with maternal metabolic complications in pregnancy (e.g., gestational diabetes, preeclampsia), but to our knowledge, there is little evidence on relationships with future metabolic risk. The aim of this study was to evaluate the association between prepregnancy obesity (prepregnancy body mass index ≥30 kg/m2) or excessive gestational weight gain (GWG; according to the 2009 recommendations from the Institute of Medicine), and maternal metabolic complications 10 y postpartum in premenopausal Chilean women. A prospective study was conducted. In 2006, 1067 Chilean mothers of children born in 2002-participants of the GOCS (Growth and Obesity Cohort Study)-were recruited. Mothers completed a questionnaire concerning sociodemographic, anthropometric, and pregnancy characteristics. Of the sample, 402 women were randomly selected to participate in a study related to the determinants of breast cancer risk in 2012. At follow-up, anthropometry, blood pressure, and fasting labs were measured. Complete data was available for 366 women. Thirty-two percent of mothers had prepregnancy overweight/obesity and 39.1% had excessive GWG. In adjusted models, prepregnancy obesity was positively associated with increased insulin resistance (odds ratio [OR], 18; 95% confidence interval [CI], 5.2-62.7), metabolic syndrome (OR, 3.3; 95% CI, 1.3-8.3), and hyperglycemia (OR, 3; 95% CI, 1.1-8.6). Prepregnancy overweight/obesity was associated with increased risk for insulin resistance, metabolic syndrome, abdominal obesity, low high-density lipoprotein cholesterol, and hypertriglyceridemia (P < 0.05). Excessive GWG was not associated with metabolic risk in the main model but was found to be positively associated in models with correction of weight by possible recall bias. Gestational obesity was associated with maternal metabolic alterations 10 y postpartum. Prevention strategies for chronic diseases should consider prepregnancy obesity as a modifiable risk factor for future metabolic health.


[Activity of the antioxidant enzymes of erythrocytes in herpetic keratitis].
Free-radical oxidations get activated, the activity of catalase slows down and the activity of superoxiddismutase increases in peripheral blood erythrocytes of patients with herpetic keratitis, which is indicative of a changing antioxidant protection in late stages of herpetic infection. The results reveal new chains in the evolution of ophthalmic herpes and confirm the feasibility of adding antioxidants to the complex therapy of patients with herpetic keratitis.


Dynamics of bacteriophage T4 presynaptic filament assembly from extrinsic fluorescence measurements of Gp32-single-stranded DNA interactions.
In the bacteriophage T4 homologous recombination system, presynaptic filament assembly on single-stranded (ssDNA) DNA requires UvsX recombinase, UvsY mediator, and Gp32 ssDNA-binding proteins. Gp32 exerts both positive and negative effects on filament assembly: positive by denaturing ssDNA secondary structure, and negative by competing with UvsX for ssDNA binding sites. UvsY is believed to help UvsX displace Gp32 from the ssDNA. To test this model we developed a real-time fluorescence assay for Gp32-ssDNA interactions during presynapsis, based on changes in the fluorescence of a 6-iodoacetamidofluorescein-Gp32 conjugate. Results demonstrate that the formation of UvsX presynaptic filaments progressively disrupts Gp32-ssDNA interactions. Under stringent salt conditions the disruption of Gp32-ssDNA by UvsX is both ATP- and UvsY-dependent. The displacement of Gp32 from ssDNA during presynapsis requires ATP binding, but not ATP hydrolysis, by UvsX protein. Likewise, UvsY-mediated presynapsis strongly requires UvsY-ssDNA interactions, and is optimal at a 1:1 stoichiometry of UvsY to UvsX and/or ssDNA binding sites. Presynaptic filaments formed in the presence of UvsY undergo assembly/collapse that is tightly coupled to the ATP hydrolytic cycle and to stringent competition for ssDNA binding sites between Gp32 and various nucleotide-liganded forms of UvsX. The data directly support the Gp32 displacement model of UvsY-mediated presynaptic filament assembly, and demonstrate that the transient induction of high affinity UvsX-ssDNA interactions by ATP are essential, although not sufficient, for Gp32 displacement. The underlying dynamics of protein-ssDNA interactions within presynaptic filaments suggests that rearrangements of UvsX, UvsY, and Gp32 proteins on ssDNA may be coupled to central processes in T4 recombination metabolism.


Kidney Ischemia/Reperfusion Injury Induces Changes in the Drug Transporter Expression at the Blood-Brain Barrier in vivo and in vitro.
Ischemia/reperfusion injury is a major cause of acute kidney injury (AKI). AKI is characterized by a sudden decrease in kidney function, systemic inflammation, oxidative stress, and dysregulation of the sodium, potassium, and water channels. While AKI leads to uremic encephalopathy, epidemiological studies have shown that AKI is associated with a subsequent risk for developing stroke and dementia. To get more insights into kidney-brain crosstalk, we have created an in vitro co-culture model based on human kidney cells of the proximal tubule (HK-2) and brain microvascular endothelial cells (BMEC). The HK-2 cell line was grown to confluence on 6-well plates and exposed to oxygen/glucose deprivation (OGD) for 4 h. Control HK-2 cells were grown under normal conditions. The BMEC cell line cerebED was grown to confluence on transwells with 0.4 μm pores. The transwell filters seeded and grown to confluence with cereEND were inserted into the plates with HK-2 cells with or without OGD treatment. In addition, cerebEND were left untreated or treated with uremic toxins, indole-3-acetic acid (IAA) and indoxyl sulfate (IS). The protein and mRNA expression of selected BBB-typical influx transporters, efflux transporters, cellular receptors, and tight junction proteins was measured in BMECs. To validate this in vitro model of kidney-brain interaction, we isolated brain capillaries from mice exposed to bilateral renal ischemia (30 min)/reperfusion injury (24 h) and measured mRNA and protein expression as described above. Both in vitro and in vivo systems showed similar changes in the expression of drug transporters, cellular receptors, and tight junction proteins. Efflux pumps, in particular Abcb1b, Abcc1, and Abcg2, have shown increased expression in our model. Thus, our in vitro co-culture system can be used to study the cellular mechanism of kidney and brain crosstalk in renal ischemia/reperfusion injury.


Characterizing VOCs emissions of coal chemical enterprise in China: a case study in five coal chemical enterprises.
Coal chemical-induced climate change has become a global concern. However, the dearth of comprehensive case studies and fundamental data has obstructed the accurate quantification of volatile organic compounds (VOCs) emissions. This has failed to equip coal chemical industries with the necessary guidelines to implement effective emission reduction strategies. In response to this, the present study meticulously examined and contrasted the VOCs emissions from five distinct coal chemical enterprises in China. This was achieved through the application of life cycle assessment (LCA), a tool used to discern the primary factors influencing VOCs emissions and to identify potential avenues for VOCs emissions reduction. The analysis revealed that BT exhibited the highest emission intensity (5.58E-04 tons/ton), followed by ED (4.89E-04 tons/ton), YL (4.23E-04 tons/ton), XJ (2.94E-04 tons/ton), and SM (1.74E-04 tons/ton). Among these enterprises, coal-to-olefins enterprises predominantly discharged VOCs via sewage treatment (average 69.12%), while coal-to-methanol enterprises primarily emitted VOCs during circulating water cooling (40.02%). In coal-to-oil enterprises, storage and blending emerged as the principal source of VOCs emissions (56.83%). As a result, this study advocates that coal chemical enterprises concentrate on curbing VOCs emissions from highly concentrated wastewater, regulating the concentration of purgeable organic carbon in circulating water cooling systems, and instituting effective treatment methods for methanol storage tank emissions. These findings proffer invaluable insights for devising VOCs control measures in regions affected by intensive coal chemical production.


The microbiome modulating activity of bile acids.
Bile acids are potent antibacterial compounds and play an important role in shaping the microbial ecology of the gut. Here, we combined flow cytometry, growth rate measurements (OD600), and NMR- and mass spectrometry-based metabolomics to systematically profile the impact of bile acids on the microbiome using in vitro and in vivo models. This study confirmed that (1) unconjugated bile acids possess more potent antibacterial activity than conjugated bile acids; (2) Gram-positive bacteria are more sensitive to bile acids than Gram-negative bacteria; (3) some probiotic bacteria such as Lactobacillus and Bifidobacterium and 7α-dehydroxylating bacteria such as Clostridium scindens show bile acid resistance that is associated with activation of glycolysis. Moreover, we demonstrated that (4) as one of most hydrophobic bile acids, lithocholic acid (LCA) shows reduced toxicity to bacteria in the cecal microbiome in both in vivo and in vitro models; (5) bile acids directly and rapidly affect bacterial global metabolism including membrane damage, disrupted amino acid, nucleotide, and carbohydrate metabolism; and (6) in vivo, short-term exposure to bile acids significantly affected host metabolism via alterations of the bacterial community structure. This study systematically profiled interactions between bile acids and gut bacteria providing validation of previous observation and new insights into the interaction of bile acids with the microbiome and mechanisms related to bile acid tolerance.


BioMeKe: an ontology-based biomedical knowledge extraction system devoted to transcriptome analysis.
Semantic interoperability between knowledge bases in medicine, and knowledge base in genomics and molecular biology will lead to advances in fundamental research as well as to improved patient care. DNA chips strategy is used for transcriptome analysis in order to identify deregulated genes in physio-pathological conditions. The objective of the BioMedical Knowledge Extraction project (BioMeKe) is to develop a knowledge warehouse in the context of transcriptome analysis during liver diseases. Knowledge sources include ontologies, related terminologies and annotations linked towards public databases (e.g., SWISSPROT). BioMeKe has been developed to have access to information using systematic investigation upon a concept, gene, gene products, pathology, or any target keyword, and is based on the combination of several relevant resources: UMLS, GeneOntology, MeSH supplementary terms, GOA, and HUGO. Current efforts are focusing on exploiting this ontology-based Knowledge Extractor, to enrich the expression data on genes delivered by a liver specific DNA microarray for better assistance of analysis.


Clozapine response trajectories and predictors of non-response in treatment-resistant schizophrenia: a chart review study.
Although clozapine is the main antipsychotic medication for treatment-resistant schizophrenia, 40-70% of patients on clozapine have persistent psychotic symptoms (i.e. ultra-treatment-resistant schizophrenia, UTRS). We aimed to examine clozapine response/non-response patterns in patients with treatment-resistant schizophrenia, as well as determine patient clinico-demographic factors associated with long-term clozapine non-response. Clinico-demographic characteristics of 241 patients on clozapine were collected through a retrospective chart review. The median (interquartile range, IQR) follow-up from illness onset was 25.0 (IQR = 24.0) years. Clozapine response was assessed at median 10.8 (IQR = 14.0) months (Time 1, T1) and 7.2 (IQR = 13.5) years (Time 2, T2) after its initiation. It was evaluated by chart reviewers based on the information provided in clinical notes. Binomial logistic regression was used to determine clinico-demographic factors associated with clozapine non-response at both T1 and T2 (i.e. stable UTRS, S-UTRS) compared to clozapine response at both times (i.e. stable clozapine responders, S-ClozResp). Among clozapine responders (n = 122) at T1, 83.6% remained clozapine responsive and 16.4% became non-responsive at T2. In the UTRS group (n = 119) at T1, 87.4% remained clozapine non-responsive and 12.6% became responsive at T2. Duration of delay in clozapine initiation (OR = 0.94, Wald χ2 = 5.33, p = 0.021) and number of pre-clozapine hospitalizations (OR = 0.95, Wald χ2 = 5.20, p = 0.023) were associated with S-UTRS. Most UTRS patients were non-responsive to clozapine from the start of treatment. Preventing delay in initiating clozapine and relapses could help promote long-term clozapine response in patients with treatment-resistant schizophrenia. Future longitudinal studies are required to explore the neuropathological correlates of relapses and delay in clozapine initiation.


The formation of the oncofetal J28 glycotope involves core-2 beta6-N-acetylglucosaminyltransferase and alpha3/4-fucosyltransferase activities.
The feto-acinar pancreatic protein or FAPP, the oncofetal glycoisoform of bile salt-dependent lipase (BSDL), is characterized by the presence of the J28 glycotope recognized by mAbJ28. This fucosylated epitope is carried out by the O-linked glycans of the C-terminal mucin-like region of BSDL. This glycotope is expressed by human tumoral pancreatic tissues and by human pancreatic tumoral cell lines such as SOJ-6 and BxPC-3 cells. However, it is not expressed by the normal human pancreatic tissues and by MiaPaCa-2 and Panc-1 cells. Due to the presence of many putative sites for O-glycosylation on FAPP and BSDL, the structure of the J28 glycotope cannot be attained by classical physical methods. In the first part of the present study, we have determined which glycosyltransferases were differently expressed in pancreatic tumoral cell lines compared to normal tissues, focusing in part on fucosyltransferases (Fuc-T) and core-2 beta6-N-acetylglucosaminyltransferase (Core2GlcNAc-T). Our data suggested that alpha2-Fuc-T activity was decreased in the four cell lines tested (SOJ-6, BxPC-3, MiaPaCa-2, and Panc-1). The alpha(1-3) and alpha(1-4) fucosylations were decreased in tumor cells that do not express the J28 glycotope whereas alpha4-Fuc-T and Core2GlcNAc-T activities were significantly increased in SOJ-6 cells which best expressed the J28 glycotope. Therefore, we wished to gain information about glycosyltransferases involved in the building of this structure by transfecting the cDNA encoding the mucin-like region of BSDL in CHO-K1 also expressing Core2GlcNAc-T and/or FUT3 and/or FUT7 activities. These CHO-K1 cells have been previously transfected with the cDNA encoding Core2GlcNAc-T and/or FUT3 and/or FUT7. Data indicated that the C-terminal peptide of BSDL (Cter) produced by those cells did not carry out the J28 glycotope unless Core2GlcNAc-T activity is present. Further transfection with FUT3 cDNA, increased the antibody recognition. Nevertheless, transfection with FUT3 or FUT7 alone did not generate the formation of the J28 glycotope on the C-terminal peptide. Furthermore, the Cter peptide produced by CHO-K1 cells expressing Core2GlcNAc-T was more reactive to the mAbJ28 after in vitro fucosylation with the recombinant soluble form of FUT3. These data suggested that the J28 glycotope encompasses structures initiated by Core2GlcNAc-T and further fucosylated by alpha3/4-Fuc-T such as FUT3, likely on GlcNAc residues.


Use of DNA repair enzymes in electrochemical detection of damage to DNA bases in vitro and in cells.
Electrochemical measurements at mercury or solid amalgam electrodes offer a highly sensitive detection of DNA strand breaks. On the other hand, electrochemical detection of damage to DNA bases at any electrode is usually much less sensitive. In this paper, we propose a new voltammetric method for the detection of the DNA base damage based on enzymatic conversion of the damaged DNA bases to single-strand breaks (ssb), single-stranded (ss) DNA regions, or both. Supercoiled DNA exposed to UV light was specifically cleaved by T4 endonuclease V, an enzyme recognizing pyrimidine dimers, the major products of photochemical DNA damage. Apurinic sites (formed in dimethyl sulfate-modified DNA) were determined after treating the DNA with E. coli exonuclease III, an enzyme introducing ssb at the abasic sites and degrading one of the DNA strands. The ssb or ssDNA regions, or both, were detected by adsorptive transfer stripping alternating current voltammetry at the mercury electrode. This technique offers much better sensitivity and selectivity of DNA base damage detection than any other electrochemical method. It is not limited to DNA damage in vitro, but it can detect also DNA base damage induced in living bacterial cells.


Optimal association-saturation procedure for estimating association and dissociation rate parameters in receptor studies. Application to solubilized A1 adenosine receptors.
A method of obtaining estimates of the maximum binding and association and dissociation rate constants for a receptor-ligand interaction is described. This new procedure, the association-saturation method, is based on an exact mathematical equation which defines the model without simplifications. The method proposed is readily applicable to any system consisting of one ligand and one receptor. With only four determinations, each involving one ligand concentration and an association time, it is possible to determine the number of specific binding sites, the dissociation and association rate constants and the equilibrium dissociation constant with accuracy. No dissociation curve is required, and only one point of the association curve for each ligand concentration is necessary. In addition to the higher confidence in the estimates of the parameter values obtained, this method leads to important savings in radiolabelled compounds and experimental time. The results were compared with those obtained with standard association-dissociation curves and saturation isotherms. The optimum number of replicates of each experimental point to obtain reliable estimates of different parameters is discussed on the basis of simulation studies. The performance of the procedure is analysed by means of association-saturation experiments with the agonist [adenine-2,8-3H,ethyl-2-3H]N6-phenylisopropyladenosine and solubilized A1 adenosine receptors from pig brain cortex.


Inhibition of glutathione synthesis increases the toxicity of oxidized low-density lipoprotein to human monocytes and macrophages.
Macrophages are thought to play an important role in the pathogenesis of atherosclerosis by mediating the oxidation of low-density lipoprotein (LDL). However, it is known that these cells show elevated glutathione levels after exposure to oxidized LDL. Here we demonstrate that this increase in the level of intracellular glutathione is due to synthesis de novo stimulated by oxidized LDL. Furthermore, inhibition of glutathione synthesis renders oxidized LDL cytotoxic to both monocytes and macrophages at a concentration well tolerated by untreated cells. The stimulation of cholesterol esterification in macrophages by low, non-toxic, concentrations of oxidized LDL is enhanced under conditions where glutathione synthesis is inhibited. These results suggest that the glutathione status of macrophages in the artery wall could be important in both controlling foam-cell formation and the detoxification of oxidized LDL.


Maintenance of a functional hematopoietic stem cell niche through galactocerebrosidase and other enzymes.
The maintenance of a functional hematopoietic niche is critical for modulating the fate of hematopoietic stem cells (HSCs). Several enzymes were described as essential for guaranteeing niche functionality. This review summarizes the recent findings about the role of galactocerebrosidase and other enzymes involved in the maintenance of a functional HSC niche. The essential role of enzymes actively involved in the maintenance of the bone marrow microenvironment, in bone remodeling, in regulating the sympathetic innervation of the niche, and in the production and relative balance of sphingolipids active in the niche has been recently highlighted. Enzymes involved in bone remodeling modify the cell-to-cell interaction between osteoblasts and HSCs. Heparanase, neutrophil elastase, and alpha-iduronidase affect the bioavailability of key cytokines and ligands within the extracellular matrix of the niche. Moreover, galactosyltransferase and galactocerebrosidase affect the function of the sympathetic nervous system and/or the balance of bioactive sphingolipids, thus influencing the SDF-1/CXCR4 axis and the proliferation of HSCs. Here, we discuss the role of different enzymes directly or indirectly influencing the niche microenvironment, and we provide a comprehensive picture of their cooperative role, together with receptors, soluble factors, and the extracellular matrix, in maintaining a functional hematopoietic niche.


Sla1, a Schizosaccharomyces pombe homolog of the human La protein, induces ectopic meiosis when its C terminus is truncated.
Sla1 is a Schizosaccharomyces pombe homolog of the human La protein. La proteins are known to be RNA-binding proteins that bear conserved RNA recognition motifs (La and RRMs), but their biological functions still have not been fully resolved. In this study, we show that the S. pombe La homolog (Sla1) is involved in regulating sexual development. Sla1 truncated in the C terminus (Sla1DeltaC) induced ectopic sporulation in the ras1Delta strain and several other sporulation-deficient mutants. The C terminus contains a nuclear localization signal. While full-length Sla1 localizes in the nucleus, Sla1DeltaC is found throughout the cell, suggesting the cytoplasmic localization of Sla1DeltaC is involved in its sporulation-inducing activity. Further deletion analysis of Sla1 indicated that a small region (35 amino acids) that includes a portion of RRM2 is sufficient to induce sporulation. The La motif (RRM1) is not involved in this activity. Strikingly, Sla1DeltaC induced haploid meiosis in a heterothallic strain, similar to the pat1-114 or mei2-SATA mutation. Sla1DeltaC induced sporulation in a mei3 disruptant but not in a mei2 disruptant, indicating that Sla1DeltaC requires Mei2 to induce haploid meiosis. Deletion of the chromosomal sla1 gene lowered the temperature sensitivity of the pat1-114 mutant. Two-hybrid analysis indicated that Pat1 interacts with Sla1DeltaC but not full-length Sla1. Thus, Sla1DeltaC may block Pat1 activity. This block would remove the inhibition on Mei2, which would then drive the cell into haploid meiosis. Finally, Sla1 was degraded prior to the start of meiosis when we monitored Sla1 in cells in which meiosis was synchronously induced. The ability of truncated Sla1 to induce ectopic meiosis represents a very novel function that has hitherto not been suspected for the La family of proteins.


Differential Functional Roles of ALDH1A1 and ALDH1A3 in Mediating Metastatic Behavior and Therapy Resistance of Human Breast Cancer Cells.
Previous studies indicate that breast cancer cells with high aldehyde dehydrogenase (ALDH) activity and CD44 expression (ALDHhiCD44⁺) contribute to metastasis and therapy resistance, and that ALDH1 correlates with poor outcome in breast cancer patients. The current study hypothesized that ALDH1 functionally contributes to breast cancer metastatic behavior and therapy resistance. Expression of ALDH1A1 or ALDH1A3 was knocked down in MDA-MB-468 and SUM159 human breast cancer cells using siRNA. Resulting impacts on ALDH activity (Aldefluor® assay); metastatic behavior and therapy response in vitro (proliferation/adhesion/migration/colony formation/chemotherapy and radiation) and extravasation/metastasis in vivo (chick choroiallantoic membrane assay) was assessed. Knockdown of ALDH1A3 but not ALDH1A1 in breast cancer cells decreased ALDH activity, and knockdown of ALDH1A1 reduced breast cancer cell metastatic behavior and therapy resistance relative to control (p < 0.05). In contrast, knockdown of ALDH1A3 did not alter proliferation, extravasation, or therapy resistance, but increased adhesion/migration and decreased colony formation/metastasis relative to control (p < 0.05). This is the first study to systematically examine the function of ALDH1 isozymes in individual breast cancer cell behaviors that contribute to metastasis. Our novel results indicate that ALDH1 mediates breast cancer metastatic behavior and therapy resistance, and that different enzyme isoforms within the ALDH1 family differentially impact these cell behaviors.


Endothelial dysfunction in a primate model of cerebral vasospasm.
Although abnormalities in the control of endothelial vasomotility have been reported in both experimental and clinical studies, the mechanism of the endothelial dysfunction that occurs following subarachnoid hemorrhage (SAH) remains unclear. Because of the absence of previous in vivo studies of endothelial function in cerebral vessels in response to SAH or cerebral vasospasm, the authors investigated endothelium-dependent responses in an established primate model of vasospasm after SAH. Endothelial function was assessed by examining vascular responses to intracarotid injections of various drugs known to act via the endothelium. Drugs that have a rapid total body clearance were selected so that their pharmacological effects would be limited to the cerebral circulation after an intracarotid infusion. Seventeen adult male cynomolgus monkeys were used. Cerebrovascular endothelium-dependent responses were examined in control animals and in animals with SAH 7, 14, and 21 days after placement of a subarachnoid clot around the right middle cerebral artery. Cortical cerebral blood flow (CBF) and cerebrovascular resistance (CVR) were recorded continuously during 5-minute intracarotid infusions of 5% dextrose vehicle, acetylcholine, histamine, bradykinin, or Calcimycin. In control animals the intracarotid infusion of acetylcholine produced a significant (7.8 +/- 9.5%) increase in CBF and a 9.3 +/- 8.7% reduction in CVR in comparison with a control infusion of dextrose vehicle. The responses to acetylcholine disappeared in animals 7 days post-SAH, specifically in the subset of animals in which arteriography confirmed the presence of vasospasm. Infusion of Calcimycin produced no significant changes in CBF or CVR in control animals, but resulted in a significant reduction in CBF and increase in CVR in animals 7 days after SAH and in animals with vasospasm. An infusion of histamine or bradykinin had no significant effect on CBF or CVR. An intracarotid infusion of acetylcholine, but not one of histamine, bradykinin, or Calcimycin, produced a measurable physiological response in the normal primate cerebrovasculature. Cerebral vasospasm that occurred after SAH produced a pathophysiological effect similar to the endothelial denudation shown in the in vitro experiments of Furchgott and Zawadzki, in which acetylcholine constricted the vessels via activation of receptors on smooth-muscle cells. Changes in vascular responses to acetylcholine and Calcimycin in animals with vasospasm, compared with control animals, provide evidence that endothelial dysfunction plays a key role in the development and/or sustenance of vasospasm after SAH.


Cognitive failures as predictors of driving errors, lapses, and violations.
Dual-process models from the cognitive literature have proposed a taxonomy of cognitive failures in everyday activities, and this novel approach was applied to understanding driver behaviour. This framework was used to examine whether categories of cognitive failure would explain driving errors, driving lapses, and driving violations in a sample of undergraduates at a large urban university. Two types of cognitive failure were examined, one associated with missing affective information and the other associated with a failure to engage effortful processes to override an automatic response. Alexithymia was used as an indicator of missing affective information, and attention regulation, reactivity, and impulsivity were used as indicators of override failure. Relevant demographic variables included gender and hours typically driven. Override failures were significantly associated with driving behaviour in the correlational analyses. In the regression analyses, attention regulation predicted driving errors, and gender, attention regulation, and impulsivity predicted driving violations. The implications of this work include the potential application to driver training, to users of informatics devices (e.g., GPS, cellular phones, messaging systems), and for individuals diagnosed with attention and/or impulsivity problems.


Management of Heart Failure in the Older Adult: Treatment and Opportunities.
OBJECTIVE: To provide a review of the available evidence regarding pharmacotherapy and areas of pharmacist intervention in transitions of care (TOC) for the geriatric population with heart failure (HF).
 DATA SOURCES: A PubMed search of articles published from 1995 through July 2018 was performed using a combination of the following words: heart failure, geriatric, elderly, (TOC), multidisciplinary, pharmacist.
 STUDY SELECTION/DATA EXTRACTION: Relevant original research, review articles, and guidelines were assessed for the management of elderly patients with HF. References from the above literature were also evaluated. Articles were selected for inclusion based on relevance to the topic, detailed methods, and complete results.
 DATA SYNTHESIS: HF is a common cause of morbidity, mortality, and hospitalizations in the elderly population. While it is important that patients adhere to evidencebased medications for HF, there are additional precautions and monitoring recommendations for this population because of a higher risk of adverse effects. Elderly patients with HF also require additional care during the transition of care process because they are at high risk for readmission during this time because of a variety of factors, including medication changes, barriers to medication use, and lack of communication between health care providers. As part of a multidisciplinary team, pharmacists can help to identify and address issues.
 CONCLUSION: Pharmacists can improve patient care outcomes in patients with HF by providing updated recommendations on pharmacotherapy and being involved in the TOC process.
.


PKCι interacts with Rab14 and modulates epithelial barrier function through regulation of claudin-2 levels.
PKCι is essential for the establishment of epithelial polarity and the normal assembly of tight junctions. We find that PKCι knockdown does not compromise the steady-state distribution of most tight junction proteins but results in increased transepithelial resistance (TER) and decreased paracellular permeability. Analysis of the levels of tight junction components demonstrates that claudin-2 protein levels are decreased. However, other tight junction proteins, such as claudin-1, ZO-1, and occludin, are unchanged. Incubation with an aPKC pseudosubstrate recapitulates the phenotype of PKCι knockdown, including increased TER and decreased levels of claudin-2. In addition, overexpression of PKCι results in increased claudin-2 levels. ELISA and coimmunoprecipitation show that the TGN/endosomal small GTPase Rab14 and PKCι interact directly. Immunolabeling shows that PKCι and Rab14 colocalize in both intracellular puncta and at the plasma membrane and that Rab14 expression is required for normal PKCι distribution in cysts in 3D culture. We showed previously that knockdown of Rab14 results in increased TER and decreased claudin-2. Our results suggest that Rab14 and aPKC interact to regulate trafficking of claudin-2 out of the lysosome-directed pathway.


Attitudes about sibling visitation in the neonatal intensive care unit.
To survey the attitudes and recommendations of staff members before and after the implementation of sibling visitation in a neonatal intensive care unit. Staff survey conducted before (1992) and after (1993) the implementation of sibling visitation. A perinatal tertiary care center. Staff members including physicians, nurses, respiratory therapists, social workers, and unit clerks (n = 139 in 1992; n = 120 in 1993). A 7-point Likert scale survey (1 = strongly disagree: 7 = strongly agree) was designed for the study. In both 1992 and 1993, the staff most strongly agreed that visitation requires special supervision, should have designated times, increases sibling knowledge, enhances sibling attachment to the baby, and increases family satisfaction. Wilcoxon rank sum tests comparing the staff across the 2 years indicated substantial attitudinal changes in favor of sibling visitation, including less perceived interference with nursing care and nursery routines (P < .01) and less concern about the infants' risk of respiratory infection and exposure to chickenpox (P < .05). There was greater attitudinal agreement between disciplines in 1993 than in 1992, suggesting better staff consensus about sibling visitation following its implementation. The recommended minimum age for visitation was 4.67 years and 4.05 years in 1992 and 1993, respectively. Brief visits of 10 to 15 minutes' duration were consistently recommended. Staff rated the sibling visitation program as successful (median = 6) on a scale ranging from 1 (very poor) to 7 (very successful). Staff members have concerns about sibling visitation that include increased risk of infection, organization, and supervision. A sibling visitation program that addresses these concerns can be successfully implemented and supported by staff, thereby fostering family-centered care in the neonatal intensive care unit.