Exclusive neuronal expression of SUCLA2 in the human brain.
SUCLA2 encodes the ATP-forming β subunit (A-SUCL-β) of succinyl-CoA ligase, an enzyme of the citric acid cycle. Mutations in SUCLA2 lead to a mitochondrial disorder manifesting as encephalomyopathy with dystonia, deafness and lesions in the basal ganglia. Despite the distinct brain pathology associated with SUCLA2 mutations, the precise localization of SUCLA2 protein has never been investigated. Here, we show that immunoreactivity of A-SUCL-β in surgical human cortical tissue samples was present exclusively in neurons, identified by their morphology and visualized by double labeling with a fluorescent Nissl dye. A-SUCL-β immunoreactivity co-localized >99 % with that of the d subunit of the mitochondrial F0-F1 ATP synthase. Specificity of the anti-A-SUCL-β antiserum was verified by the absence of labeling in fibroblasts from a patient with a complete deletion of SUCLA2. A-SUCL-β immunoreactivity was absent in glial cells, identified by antibodies directed against the glial markers GFAP and S100. Furthermore, in situ hybridization histochemistry demonstrated that SUCLA2 mRNA was present in Nissl-labeled neurons but not glial cells labeled with S100. Immunoreactivity of the GTP-forming β subunit (G-SUCL-β) encoded by SUCLG2, or in situ hybridization histochemistry for SUCLG2 mRNA could not be demonstrated in either neurons or astrocytes. Western blotting of post mortem brain samples revealed minor G-SUCL-β immunoreactivity that was, however, not upregulated in samples obtained from diabetic versus non-diabetic patients, as has been described for murine brain. Our work establishes that SUCLA2 is expressed exclusively in neurons in the human cerebral cortex.


Infiltration of the synovial membrane with macrophage subsets and polymorphonuclear cells reflects global disease activity in spondyloarthropathy.
Considering the relation between synovial inflammation and global disease activity in rheumatoid arthritis (RA) and the distinct but heterogeneous histology of spondyloarthropathy (SpA) synovitis, the present study analyzed whether histopathological features of synovium reflect specific phenotypes and/or global disease activity in SpA. Synovial biopsies obtained from 99 SpA and 86 RA patients with active knee synovitis were analyzed for 15 histological and immunohistochemical markers. Correlations with swollen joint count, serum C-reactive protein concentrations, and erythrocyte sedimentation rate were analyzed using classical and multiparameter statistics. SpA synovitis was characterized by higher vascularity and infiltration with CD163+ macrophages and polymorphonuclear leukocytes (PMNs) and by lower values for lining-layer hyperplasia, lymphoid aggregates, CD1a+ cells, intracellular citrullinated proteins, and MHC-HC gp39 complexes than RA synovitis. Unsupervised clustering of the SpA samples based on synovial features identified two separate clusters that both contained different SpA subtypes but were significantly differentiated by concentration of C-reactive protein and erythrocyte sedimentation rate. Global disease activity in SpA correlated significantly with lining-layer hyperplasia as well as with inflammatory infiltration with macrophages, especially the CD163+ subset, and with PMNs. Accordingly, supervised clustering using these synovial parameters identified a cluster of 20 SpA patients with significantly higher disease activity, and this finding was confirmed in an independent SpA cohort. However, multiparameter models based on synovial histopathology were relatively poor predictors of disease activity in individual patients. In conclusion, these data indicate that inflammatory infiltration of the synovium with CD163+ macrophages and PMNs as well as lining-layer hyperplasia reflect global disease activity in SpA, independently of the SpA subtype. These data support a prominent role for innate immune cells in SpA synovitis and warrant further evaluation of synovial histopathology as a surrogate marker in early-phase therapeutic trials in SpA.


Cerebral vascular adaptation to pregnancy and its role in the neurological complications of eclampsia.
The cerebral circulation has a central role in mediating the neurological complications of eclampsia, yet our understanding of how pregnancy and preeclampsia affect this circulation is severely limited. Here, we show that pregnancy causes outward remodeling of penetrating arterioles and increased capillary density in the brain due to activation of peroxisome proliferator-activated receptor-γ (PPARγ), a transcription factor involved in cerebrovascular remodeling and highly activated in pregnancy. Pregnancy-induced PPARγ activation also significantly affected cerebral hemodynamics, decreasing vascular resistance and increasing cerebral blood flow by ∼40% in response to acute hypertension that caused breakthrough of autoregulation. These structural and hemodynamic changes in the brain during pregnancy were associated with substantially increased blood-brain barrier permeability, an effect that could promote passage of damaging proteins into the brain and cause the neurological complications of eclampsia, including seizure.


General adherence to guideline recommendations on initial diagnosis of bladder cancer in the United States and influencing factors.
Because international guidelines recommend best practices regarding staging of incident bladder cancer, we determined the adherence to such recommendations in the United States, performing a large retrospective database analysis. Patients with the diagnosis of urothelial cancer were identified in the Surveillance, Epidemiology, and End Results (SEER)-Medicare database between 1992 and 2007. Staging procedures were identified and analyzed. As reference for published recommendations, we used the American Urological Association (AUA), European Association of Urology (EAU), and National Comprehensive Cancer Network (NCCN) guidelines. Based on these sources, recommended initial staging of bladder cancer was analyzed. Of all 56,130 patients, 6148 (10.9%) had a cytologic examination, 29,677 (52.9%) had a standard urinalysis, 2882 (5.1%) underwent intravenous pyelography (IVP), 6950 (12.4%) underwent retrograde pyelography (RPG), and 8145 (14.5%) had computed tomography/magnetic resonance imaging (CT/MRI). There was a significant trend over the years to a higher use of cytologic analysis, standard urinalysis, and CT/MRI. We observed a significant trend toward a lower rate of IVP and a stable use of RPG. The limitation of our study is that claims data are designed for payment processing, not quality measurement. Despite published recommendations on the initial diagnosis of bladder cancer, our data show that less than half of the included patients received all the elements thought to be required for an initial diagnosis of bladder cancer as recommended by guidelines. Greater adherence to recommendations may ensure optimal treatment strategies. Appropriate treatment is critical to patient outcomes, because evidence-based therapeutic management can be practiced only if an accurate assessment of the disease takes place at the time of initial diagnosis.


Sustained effects of prior red light on pupil diameter and vigilance during subsequent darkness.
Environmental light can exert potent effects on physiology and behaviour, including pupil size, vigilance and sleep. Previous work showed that these non-image forming effects can last long beyond discontinuation of short-wavelength light exposure. The possible functional effects after switching off long-wavelength light, however, have been insufficiently characterized. In a series of controlled experiments in healthy adult volunteers, we evaluated the effects of five minutes of intense red light on physiology and performance during subsequent darkness. As compared to prior darkness, prior red light induced a subsequent sustained pupil dilation. Prior red light also increased subsequent heart rate and heart rate variability when subjects were asked to perform a sustained vigilance task during the dark exposure. While these changes suggest an increase in the mental effort required for the task, it could not prevent a post-red slowing of response speed. The suggestion that exposure to intense red light affects vigilance during subsequent darkness, was confirmed in a controlled polysomnographic study that indeed showed a post-red facilitation of sleep onset. Our findings suggest the possibility of using red light as a nightcap.


Preliminary development of humanistic care indicators for residents in nursing homes: a Delphi technique.
The overwhelming majority of residents among nursing homes are the elderly in Taiwan. Previous studies have shown the dissatisfaction with care from the viewpoints of nursing home residents. For improving the care quality of nursing homes, the study aims to develop humanistic care indicators (HCIs). The Delphi technique was used to develop the HCIs through the consensus of 23 experts. Through three rounds of questionnaires, the expert panel reached a consensus. Forty-four HCIs for nursing home were identified and grouped into eight elements: friendly environment, holistic care, empathy, individualization, autonomy, decision-making participation, appropriate use of tools, and serious assessment of customer opinion. This study compiled related literature and conducted a Delphi survey to transform humanistic care from an abstract concept into concrete indicators for evaluation. These findings could serve as a guideline for the care providers in nursing homes. Further studies are needed to test the practicability of HCIs and evaluate the outcomes of applying HCIs in nursing homes.


Effects of hydrogen-rich saline on rats with acute carbon monoxide poisoning.
Studies have shown that inhalation of hydrogen gas, which acts as an antioxidant, can protect the brain against free radicals in rats with ischemia-reperfusion. The neuronal damage caused by acute carbon monoxide (CO) poisoning is partly free radical mediated. We hypothesize that hydrogen may prevent neurological damage from CO poisoning. This study is designed to test whether hydrogen (H(2))-rich saline will have a protective effect on rats with acute CO poisoning. Male Sprague-Dawley rats were subjected to CO poisoning. H(2)-rich saline was administered by peritoneal injection (6 mL/kg/24 h). We used the Morris water maze and the open field test to determine cognitive function. After cognitive function studies, rats were decapitated and the levels of trace elements copper (Cu), zinc (Zn), and iron (Fe) in serum and brain were assessed by flame atomic absorption spectrometry. Necrosis, apoptosis, and autophagy of neurons were assessed by H-E staining and immunohistochemical staining in another group of rats. H(2)-rich saline treatment improved the cognitive deficits and reduced the degree of necrosis, apoptosis, and cell autophagy in rats. Additionally, H(2)-rich saline decreased the content of Fe in serum and brain in these rats, and increased the content of serum Cu related to free radical metabolism. H(2)-rich saline may effectively protect the brain from injury after acute CO poisoning. The mechanism of this protection may be related to lessening oxidative damage by affecting trace elements in vivo.


Phosphatidylcholine's functions beyond that of a membrane brick.
Since its discovery in the 19th century, phosphatidylcholine (PC) has been regarded primarily as a structural lipid. However, more recent evidence, much of it in the last five years, strongly suggests that PC has other roles. Here, we explore some of that new evidence and consider the possibility that the ultimate role of phosphatidylcholine may not be predictable.


VGLUT2-dependent sensory neurons in the TRPV1 population regulate pain and itch.
The natural response to itch sensation is to scratch, which relieves the itch through an unknown mechanism. Interaction between pain and itch has been frequently demonstrated, and the selectivity hypothesis of itch, based on data from electrophysiological and behavioral experiments, postulates the existence of primary pain afferents capable of repressing itch. Here, we demonstrate that deletion of vesicular glutamate transporter (VGLUT) 2 in a subpopulation of neurons partly overlapping with the vanilloid receptor (TRPV1) primary afferents resulted in a dramatic increase in itch behavior accompanied by a reduced responsiveness to thermal pain. The increased itch behavior was reduced by administration of antihistaminergic drugs and by genetic deletion of the gastrin-releasing peptide receptor, demonstrating a dependence on VGLUT2 to maintain normal levels of both histaminergic and nonhistaminergic itch. This study establishes that VGLUT2 is a major player in TRPV1 thermal nociception and also serves to regulate a normal itch response.


A Next-Generation Sequencing-Based Molecular Approach to Characterize a Tick Vector in Lyme Disease.
Next-generation sequencing approaches have revolutionized genomic medicine and enabled rapid diagnosis for several diseases. These approaches are widely used for pathogen detection in several infectious diseases. Lyme disease is a tick-borne infectious disease, which affects multiple organs. The causative organism is a spirochete, Borrelia burgdorferi, which is transmitted by ticks. Lyme disease can be treated easily if detected early, but its diagnosis is often delayed or is incorrect leading to a chronic debilitating condition. Current confirmatory diagnostic tests for Lyme disease rely on detection of antigens derived from B. burgdorferi, which are prone to both false positives and false negatives. Instead of focusing only on the human host for the diagnosis of Lyme disease, one could also attempt to identify the vector (tick) and the causative organism carried by the tick. Since all ticks do not transmit Lyme disease, it can be informative to accurately identify the tick from the site of bite, which is often observed by the patient and discarded. However, identifying ticks based on morphology alone requires a trained operator and can still be incorrect. Thus, we decided to take a molecular approach by sequencing DNA and RNA from a tick collected from an individual bitten by the tick. Using next-generation sequencing, we confirmed the identity of the tick as a dog tick, Dermacentor variabilis, and did not identify any pathogenic bacterial sequences, including Borrelia species. Despite the limited availability of nucleotide sequences for many types of ticks, our approach correctly identified the tick species. This proof-of-principle study demonstrates the potential of next-generation sequencing in the diagnosis of tick-borne infections, which can also be extended to other zoonotic diseases.


A Phase I Trial of Combined Ridaforolimus and MK-2206 in Patients with Advanced Malignancies.
The PI3K/Akt/mTOR signaling pathway is aberrantly activated in many cancers. Combining ridaforolimus, an mTOR inhibitor, with MK-2206, an Akt inhibitor, may more completely block the PI3K pathway and inhibit tumor growth. This phase I study assessed dose-limiting toxicities (DLT) and maximum tolerated dose (MTD) for the combination of oral ridaforolimus plus oral MK-2206 in patients with advanced solid tumors. Efficacy was evaluated in patients with biomarker-identified estrogen receptor-positive breast cancer (low RAS gene signature and high Ki67 index) or castration-resistant prostate cancer (PTEN deficiency) with PI3K pathway addiction. Thirty-five patients were enrolled: 11 patients in part A (three breast cancer) and 24 biomarker-eligible patients in part B (16 breast cancer, eight prostate cancer). One patient with breast cancer from part A was also found to be biomarker-eligible when tested after she had clinical response. The MTD was 10 mg/d ridaforolimus 5 d/wk + 90 mg/wk MK-2206; 1 of 17 patients experienced DLT (grade 3 rash) at this dose. The most common adverse events at MTD were rash (44.4%), stomatitis (38.9%), diarrhea (27.8%), and decreased appetite (27.8%). By investigator assessment, 2 of 16 (12.5%) evaluable patients with breast cancer had partial response; by central assessment, 2 of 14 (14.3%) evaluable patients had complete response. Two patients had durable stable disease (SD) for 416 and 285 days, respectively. No patients with prostate cancer responded; one patient had SD for ≥ 6 months. Combination ridaforolimus and MK-2206 showed promising activity and good tolerability in heavily pretreated patients with hormone-positive and -negative breast cancer exhibiting PI3K pathway dependence.


In vivo genome editing improves motor function and extends survival in a mouse model of ALS.
Amyotrophic lateral sclerosis (ALS) is a fatal and incurable neurodegenerative disease characterized by the progressive loss of motor neurons in the spinal cord and brain. In particular, autosomal dominant mutations in the superoxide dismutase 1 (SOD1) gene are responsible for ~20% of all familial ALS cases. The clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated (Cas9) genome editing system holds the potential to treat autosomal dominant disorders by facilitating the introduction of frameshift-induced mutations that can disable mutant gene function. We demonstrate that CRISPR-Cas9 can be harnessed to disrupt mutant SOD1 expression in the G93A-SOD1 mouse model of ALS following in vivo delivery using an adeno-associated virus vector. Genome editing reduced mutant SOD1 protein by >2.5-fold in the lumbar and thoracic spinal cord, resulting in improved motor function and reduced muscle atrophy. Crucially, ALS mice treated by CRISPR-mediated genome editing had ~50% more motor neurons at end stage and displayed a ~37% delay in disease onset and a ~25% increase in survival compared to control animals. Thus, this study illustrates the potential for CRISPR-Cas9 to treat SOD1-linked forms of ALS and other central nervous system disorders caused by autosomal dominant mutations.


Localization of cholinergic and purinergic receptors on outer hair cells isolated from the guinea-pig cochlea.
Acetylcholine (ACh) and adenosine 5'-triphosphate (ATP) are shown to act in opposing fashion on guinea-pig cochlear outer hair cells (OHCS) via receptors localized within different fluid compartments of the organ of Corti. The cholinergic (efferent) receptors localized at the basal (perilymphatic) region of these cells activated a rapidly desensitizing hyperpolarizing K+ current. In contrast, purinergic (ATP) receptors were localized at the apical (endolymphatic) surface of OHCS and activated a depolarizing nonselective cation current which exhibited inward rectification and lacked desensitization. Localization of the receptors was determined by using whole-cell patch-clamp, by recording onset latencies and response amplitudes to pulses of either ACh or ATP pressure-applied at selected sites along the length of isolated OHCS. Under voltage-clamp at -60 mV, the largest ACh-induced (outward) currents were recorded when ACh was directed at the basal region of the cells. Conversely, the maximum (inward) ATP currents were obtained when ATP was directed toward the apical surface of these cells. Onset latencies increased rapidly from a minimum of approximately 10 ms for either ACh or ATP as the drug pipette was moved away from these optimal sites. The ATP response was antagonized by amiloride in a dose-dependent manner with a KD of approximately 400 microM. The localization of P2-type purinoceptors to the endolymphatic surface of OHCS suggests that ATP mediates a humoral modulation of the mechano-electrical transduction process.


Substrate recognition of holocytochrome c synthase: N-terminal region and CXXCH motif of mitochondrial cytochrome c.
Holocytochrome c synthase (HCCS) attaches heme covalently to mitochondrial respiratory cytochromes c. Little is known about the reaction of heme attachment to apocytochromes c by HCCS, although recently it has been established that the CXXCH motif and the N-terminus of the apocytochrome polypeptide are important protein-protein recognition motifs. Here, we explore further the important features of the N-terminal sequence and investigate what variations in the CXXCH residues are productively recognised by HCCS in its substrate.


The Emerging Role of Chromatin Remodeling Factors in Female Pubertal Development.
To attain sexual competence, all mammalian species go through puberty, a maturational period during which body growth and development of secondary sexual characteristics occur. Puberty begins when the diurnal pulsatile gonadotropin-releasing hormone (GnRH) release from the hypothalamus increases for a prolonged period of time, driving the adenohypophysis to increase the pulsatile release of luteinizing hormone with diurnal periodicity. Increased pubertal GnRH secretion does not appear to be driven by inherent changes in GnRH neuronal activity; rather, it is induced by changes in transsynaptic and glial inputs to GnRH neurons. We now know that these changes involve a reduction in inhibitory transsynaptic inputs combined with increased transsynaptic and glial excitatory inputs to the GnRH neuronal network. Although the pubertal process is known to have a strong genetic component, during the last several years, epigenetics has been implicated as a significant regulatory mechanism through which GnRH release is first repressed before puberty and is involved later on during the increase in GnRH secretion that brings about the pubertal process. According to this concept, a central target of epigenetic regulation is the transcriptional machinery of neurons implicated in stimulating GnRH release. Here, we will briefly review the hormonal changes associated with the advent of female puberty and the role that excitatory transsynaptic inputs have in this process. In addition, we will examine the 3 major groups of epigenetic modifying enzymes expressed in the neuroendocrine hypothalamus, which was recently shown to be involved in pubertal development and progression.


Bioethical Issues in Biostatistical Consulting Study: Additional Findings and Concerns.
To determine the frequency and severity of 17 requests for inappropriate analysis and reporting of data that biostatisticians received from researchers in statistical consultations, as indirectly experienced (i.e., either heard about or observed being asked of other biostatisticians). A randomly drawn sample of 522 members of the American Statistical Association who self-identified as consulting biostatisticians were selected to participate in an online survey with the Bioethical Issues in Biostatistical Consulting Questionnaire, specifically developed for this study. Of the 522 consulting biostatisticians contacted, 390 (74.7%) completed the survey. The top 4 most frequently reported indirectly experienced inappropriate requests rated as "high severity" by at least 50% of the respondents were as follows: 1) proposing a study with a flawed design, including insufficient power; 2) setting aside values when the outcome turns on a few outliers; 3) reporting results of data analysis from only subsets of the data; and 4) overstating the statistical findings well beyond what the data support so that readers are misled. This article is a follow-up report to our recently published article on 18 directly experienced inappropriate and/or unethical requests by biomedical researchers of their consulting biostatisticians. These additional survey findings from the Bioethical Issues in Biostatistical Consulting Questionnaire on 17 indirectly experienced inappropriate requests show the following: 1) the widespread nature of inappropriate, if not, frankly unethical requests made by biomedical researchers of their consulting biostatisticians and 2) the urgent need to develop and offer educational programs for biomedical researchers in training to correct these behaviors-be they intentional or accidental. This report speaks to the urgency for developing training programs for new and existing researchers to reduce the frequency of these inappropriate bioethical requests during biostatistical consultations. The findings from this U.S. national survey of biostatisticians on inappropriate requests for data analyses by biomedical researchers strongly suggest a need for remedial actions, including 1) new educational modules for in-development and currently employed biomedical researchers, 2) improved institutional environments by research universities and companies regarding job and publication pressures, and 3) inclusion of a collaborating biostatistician as a working research team member from the early planning stages of all biomedical studies.


Recurrent missense variants in clonal hematopoiesis-related genes present in the general population.
Clonal hematopoiesis (CH) consists in an abnormal expansion of a hematopoietic stem cell bearing an advantageous somatic variant. A survey of known recurrent somatic missense variants in DNMT3A, SF3B1, SRSF2, and TP53, some of the most prominent genes underlying CH of indeterminate potential (CHIP), in gnomAD noncancer database shows the presence of 73 variants. Many of them reach frequencies higher than 0.01% in various populations and, in many cases, are enriched in specific populations. Consistent with a potential involvement in CHIP, we found that the age distribution of the carriers is shifted towards old ages. Moreover, the variant allele frequencies are on average lower than 50%, expected for germline heterozygous variants. The pervasive presence of some of such variants in blood DNA from elder individuals is compatible with CHIP of somatic origin. On practical grounds, CHIP can lead to misclassification of somatic variants in cancer-predisposition genes as inherited, which bear consequences for the affected individuals and their families.


Deletion of Kncn Does Not Affect Kinocilium and Stereocilia Bundle Morphogenesis and Mechanotransduction in Cochlear Hair Cells.
Auditory hair cells possess stunning cilia structure that composes of a bundle of stereocilia for mechano-electrical transduction and a single kinocilium for guiding the polarity of hair bundle towards maturation. However, the molecules underlying kinocilium function have not yet been fully understood. Hence, the proteins involved in hair bundle development and function are of a large interest. From a fine microarray analysis, we found that kinocilin (Kncn) was enriched in hair cell specific expression profile. Consistently, it has been reported that KNCN was a protein mainly located in the kinocilium of hair cells in the inner ear. However, the hypothesis that KNCN is a kinocilium protein has not been validated in mice with Kncn gene perturbed. In this study, we generated Kncn knockout mouse lines by CRISPR/Cas9 technique and further examined the morphology and function of cochlear hair cells. Our results showed that there was no obvious hearing loss in the knockout mice, determined by audiometry. Histological study demonstrated that the inner ear and hair cell structure were intact. Especially, there was no deficit of mechanotransduction (MET) in cochlear outer hair cells (OHCs). In summary, our work suggests that KNCN is not essential for kinocilium-oriented hair bundle function in cochlear hair cells.


Dimethyl Fumarate Inhibits the Nuclear Factor κB Pathway in Breast Cancer Cells by Covalent Modification of p65 Protein.
In breast tumors, activation of the nuclear factor κB (NFκB) pathway promotes survival, migration, invasion, angiogenesis, stem cell-like properties, and resistance to therapy--all phenotypes of aggressive disease where therapy options remain limited. Adding an anti-inflammatory/anti-NFκB agent to breast cancer treatment would be beneficial, but no such drug is approved as either a monotherapy or adjuvant therapy. To address this need, we examined whether dimethyl fumarate (DMF), an anti-inflammatory drug already in clinical use for multiple sclerosis, can inhibit the NFκB pathway. We found that DMF effectively blocks NFκB activity in multiple breast cancer cell lines and abrogates NFκB-dependent mammosphere formation, indicating that DMF has anti-cancer stem cell properties. In addition, DMF inhibits cell proliferation and significantly impairs xenograft tumor growth. Mechanistically, DMF prevents p65 nuclear translocation and attenuates its DNA binding activity but has no effect on upstream proteins in the NFκB pathway. Dimethyl succinate, the inactive analog of DMF that lacks the electrophilic double bond of fumarate, is unable to inhibit NFκB activity. Also, the cell-permeable thiol N-acetyl l-cysteine, reverses DMF inhibition of the NFκB pathway, supporting the notion that the electrophile, DMF, acts via covalent modification. To determine whether DMF interacts directly with p65, we synthesized and used a novel chemical probe of DMF by incorporating an alkyne functionality and found that DMF covalently modifies p65, with cysteine 38 being essential for the activity of DMF. These results establish DMF as an NFκB inhibitor with anti-tumor activity that may add therapeutic value in the treatment of aggressive breast cancers.


Prescription and Other Medication Use in Pregnancy.
To characterize prescription and other medication use in a geographically and ethnically diverse cohort of women in their first pregnancy. In a prospective, longitudinal cohort study of nulliparous women followed through pregnancy from the first trimester, medication use was chronicled longitudinally throughout pregnancy. Structured questions and aids were used to capture all medications taken as well as reasons they were taken. Total counts of all medications taken including number in each category and class were captured. Additionally, reasons the medications were taken were recorded. Trends in medications taken across pregnancy and in the first trimester were determined. Of the 9,546 study participants, 9,272 (97.1%) women took at least one medication during pregnancy with 9,139 (95.7%) taking a medication in the first trimester. Polypharmacy, defined as taking at least five medications, occurred in 2,915 (30.5%) women. Excluding vitamins, supplements, and vaccines, 73.4% of women took a medication during pregnancy with 55.1% taking one in the first trimester. The categories of drugs taken in pregnancy and in the first trimester include the following: gastrointestinal or antiemetic agents (34.3%, 19.5%), antibiotics (25.5%, 12.6%), and analgesics (23.7%, 15.6%, which includes 3.6%; 1.4% taking an opioid pain medication). In this geographically and ethnically diverse cohort of nulliparous pregnant women, medication use was nearly universal and polypharmacy was common. ClinicalTrials.gov, NCT01322529.


A new bismuth iron oxyphosphate,Bi6(Bi0.32Fe0.68)(PO4)4O4.
Iron was inserted into the known crystal structure of the bismuth phosphate oxide Bi(6.67)(PO(4))(4)O(4) to ascertain its location in the vacancies associated with the bismuth ion located at the origin of the unit cell. Single-crystal X-ray diffraction refinements converged to a model of composition Bi(6)(Bi(0.32)Fe(0.68))(PO(4))(4)O(4) (hexabismuth iron tetraphosphate tetraoxide), in which Bi and Fe are displaced from the origin giving rise to a random distribution over the 2i sites instead of 1a, the origin of space group P1. The isotropic displacement parameter for Bi/Fe has a reasonable value in this model. This structure establishes for the first time that Fe substitutes in the Bi-deficient site in this series of materials and that Fe and Bi are disordered around the center of symmetry. These results enhance understanding of the crystal chemistry of these main group phosphates that are of interest in ion transport.


The antidiuretic effect of chronic hydrochlorothiazide treatment in rats with diabetes insipidus: water and electrolyte balance.
1. The antidiuretic effect of hydrochlorothiazide in diabetes insipidus was investigated in rats with the hereditary hypothalamic form of the disease (Brattleboro rats). 2. Administration of hydrochlorothiazide in the food resulted in a marked fall in urine volume and a corresponding rise in osmolality. These effects persisted throughout the period of treatment (6-7 days). 3. Body weight and extracellular volume were significantly reduced in the thiazide-treated rats. 4. Hydrochlorothiazide caused an increase in urinary sodium excretion only on the first day of treatment. The resulting small negative sodium balance (in comparison with untreated rats) remained statistically significant for 2 days only. Thiazide-treated rats gradually developed a potassium deficit which was statistically significant from the fourth day of treatment. 5. Total exchangeable sodium, measured after 7 days of thiazide treatment, was not significantly different from that of untreated rats. However, plasma sodium was reduced in thiazide-treated animals, whereas erythrocyte sodium concentration was elevated. 6. It is concluded that the antidiuresis resulting from chronic hydrochlorothiazide administration is associated with a reduction in extracellular volume, but not with a significant overall sodium deficit. Hydrochlorothiazide appears to cause a redistribution of the body's sodium such that the amount of sodium in the extracellular fluid compartment is reduced.


Functional and transcriptional characterization of human embryonic stem cell-derived endothelial cells for treatment of myocardial infarction.
Differentiation of human embryonic stem cells into endothelial cells (hESC-ECs) has the potential to provide an unlimited source of cells for novel transplantation therapies of ischemic diseases by supporting angiogenesis and vasculogenesis. However, the endothelial differentiation efficiency of the conventional embryoid body (EB) method is low while the 2-dimensional method of co-culturing with mouse embryonic fibroblasts (MEFs) require animal product, both of which can limit the future clinical application of hESC-ECs. Moreover, to fully understand the beneficial effects of stem cell therapy, investigators must be able to track the functional biology and physiology of transplanted cells in living subjects over time. In this study, we developed an extracellular matrix (ECM) culture system for increasing endothelial differentiation and free from contaminating animal cells. We investigated the transcriptional changes that occur during endothelial differentiation of hESCs using whole genome microarray, and compared to human umbilical vein endothelial cells (HUVECs). We also showed functional vascular formation by hESC-ECs in a mouse dorsal window model. Moreover, our study is the first so far to transplant hESC-ECs in a myocardial infarction model and monitor cell fate using molecular imaging methods. Taken together, we report a more efficient method for derivation of hESC-ECs that express appropriate patterns of endothelial genes, form functional vessels in vivo, and improve cardiac function. These studies suggest that hESC-ECs may provide a novel therapy for ischemic heart disease in the future.


Yeast meiotic mutants proficient for the induction of ectopic recombination.
A screen was designed to identify Saccharomyces cerevisiae mutants that were defective in meiosis yet proficient for meiotic ectopic recombination in the return-to-growth protocol. Seven mutants alleles were isolated; two are important for chromosome synapsis (RED1, MEK1) and five function independently of recombination (SPO14, GSG1, SPOT8/MUM2, 3, 4). Similar to the spoT8-1 mutant, mum2 deletion strains do not undergo premeiotic DNA synthesis, arrest prior to the first meiotic division and fail to sporulate. Surprisingly, although DNA replication does not occur, mum2 mutants are induced for high levels of ectopic recombination. gsg1 diploids are reduced in their ability to complete premeiotic DNA synthesis and the meiotic divisions, and a small percentage of cells produce spores. mum3 mutants sporulate poorly and the spores produced are inviable. Finally, mum4-1 mutants produce inviable spores. The meiotic/sporulation defects of gsg1, mum2, and mum3 are not relieved by spo11 or spo13 mutations, indicating that the mutant defects are not dependent on the initiation of recombination or completion of both meiotic divisions. In contrast, the spore inviability of the mum4-1 mutant is rescued by the spo13 mutation. The mum4-1 spo13 mutant undergoes a single, predominantly equational division, suggesting that MUM4 functions at or prior to the first meiotic division. Although recombination is variably affected in the gsg1 and mum mutants, we hypothesize that these mutants define genes important for aspects of meiosis not directly related to recombination.


The epidemiology of infectious diseases of livestock.
From the time of the first modern studies of infectious diseases, by Koch, Pasteur, Theiler and their colleagues, it has been clear that laboratory investigation must be complemented by epidemiologic investigation. The measurement of all aspects of the natural history of a disease in naturally affected populations is necessary if we are to rationally design control regimens. Building upon a historic perspective, this paper presents a view of the present status of epidemiology as it pertains to animal disease control, and presents a view of the merits of expanding the use of this science in future animal disease control programs, internationally, in developed and developing countries. The basis for this view lies in adaptation of principles employed in human infectious disease epidemiology, and principles which guide the organization of international disease control agencies.


The responsiveness of quality of life utilities to change in depression: a comparison of instruments (SF-6D, EQ-5D, and DFD).
Utilities are often a main outcome parameter in economic evaluations. Because depression has a large influence on quality of life, it is expected that utilities are responsive to changes in depression. To evaluate the change in utility derived from different instruments in depression, including the Short Form 6D (SF-6D), the Euroqol based on the UK (EQ-5D(UK)), the Euroqol based on the Dutch tariff (EQ-5D(NL)), and utilities derived from Beck Depression Inventory Second Edition (BDI-II) using the Depression-Free-Day method. This study evaluated the responsiveness, the minimally important difference, and the agreement in utility change derived from the different instruments. The SF-6D, EQ-5D(UK), and EQ-5D(NL) were responsive. The minimally important difference values are in line with previous studies, about 0.3. The Depression-Free-Day method nearly always resulted in positive utility changes, even for subgroups that had no change or deterioration in health status or depression. There was poor agreement between utility changes of the SF-6D, EQ-5D (either EQ-5D(UK) or EQ-5D(NL)), and DFDu. The SF-6D, EQ-5D(UK), and EQ-5D(NL) seem responsive and thus adequate for estimating utility in depression treatment. We do not recommend the use of the Depression-Fee-Day method. The low agreement between utility changes indicates that outcomes of the different instruments are incomparable.


Complete genome determination and analysis of Acholeplasma oculi strain 19L, highlighting the loss of basic genetic features in the Acholeplasmataceae.
Acholeplasma oculi belongs to the Acholeplasmataceae family, comprising the genera Acholeplasma and 'Candidatus Phytoplasma'. Acholeplasmas are ubiquitous saprophytic bacteria. Several isolates are derived from plants or animals, whereas phytoplasmas are characterised as intracellular parasitic pathogens of plant phloem and depend on insect vectors for their spread. The complete genome sequences for eight strains of this family have been resolved so far, all of which were determined depending on clone-based sequencing. The A. oculi strain 19L chromosome was sequenced using two independent approaches. The first approach comprised sequencing by synthesis (Illumina) in combination with Sanger sequencing, while single molecule real time sequencing (PacBio) was used in the second. The genome was determined to be 1,587,120 bp in size. Sequencing by synthesis resulted in six large genome fragments, while the single molecule real time sequencing approach yielded one circular chromosome sequence. High-quality sequences were obtained by both strategies differing in six positions, which are interpreted as reliable variations present in the culture population. Our genome analysis revealed 1,471 protein-coding genes and highlighted the absence of the F1FO-type Na+ ATPase system and GroEL/ES chaperone. Comparison of the four available Acholeplasma sequences revealed a core-genome encoding 703 proteins and a pan-genome of 2,867 proteins. The application of two state-of-the-art sequencing technologies highlights the potential of single molecule real time sequencing for complete genome determination. Comparative genome analyses revealed that the process of losing particular basic genetic features during genome reduction occurs in both genera, as indicated for several phytoplasma strains and at least A. oculi. The loss of the F1FO-type Na+ ATPase system may separate Acholeplasmataceae from other Mollicutes, while the loss of those genes encoding the chaperone GroEL/ES is not a rare exception in this bacterial class.


Structural insights into the loss of penicillinase and the gain of ceftazidimase activities by OXA-145 β-lactamase in Pseudomonas aeruginosa.
We previously described extended-spectrum oxacillinase OXA-145 from Pseudomonas aeruginosa, which differs from narrow-spectrum OXA-35 by loss of Leu-155. The deletion results in loss of benzylpenicillin hydrolysis and acquisition of activity against ceftazidime. We report the crystal structure of OXA-145 and provide the basis of its switch in substrate specificity. OXA-145 variants were generated by site-directed mutagenesis and purified to homogeneity. The crystal structure of OXA-145 was determined and molecular dynamics simulations were performed. Kinetic parameters were investigated in the absence and in the presence of sodium hydrogen carbonate (NaHCO3) for representative substrates. The structure of OXA-145 was obtained at a resolution of 2.3 Å and its superposition with that of OXA-10 showed that Trp-154 was shifted by 1.8 Å away from the catalytic Lys-70, which was not N-carboxylated. Addition of NaHCO3 significantly increased the catalytic efficiency against penicillins, but not against ceftazidime. The active-site cavity of OXA-145 was larger than that of OXA-10, which may favour the accommodation of large molecules such as ceftazidime. Molecular dynamics simulations of OXA-145 in complex with ceftazidime revealed two highly coordinated water molecules on the α- or β-face of the acyl ester bond, between Ser-67 and ceftazidime, which could be involved in the catalytic process. Deletion of Leu-155 resulted in inefficient positioning of Trp-154, leading to a non-carboxylated Lys-70 and thus to loss of hydrolysis of the penicillins. Ceftazidime hydrolysis could be attributed to enlargement of the active site and to a catalytic mechanism independent of the carboxylated Lys-70.


Conformational constraints in nuclear DNA.
We have investigated DNA superstructure in a wide range of nuclei of higher cells by gently lysing cells to release structures that resemble nuclei but are depleted of nuclear proteins. The sedimentation properties of these structures, which we call nucleoids, have been examined in sucrose gradients containing the intercalating agent, ethidium. The sedimentation rate of nucleoids derived from the growing cells of mammals, birds, amphibians and insects varies in the manner characteristic of circular and superhelical molecules of DNA. These characteristic changes in sedimentation rate are abolished by irradiating the nucleoids with low doses of gamma-rays, a procedure known to introduce single-strand scissions into DNA. We have also investigated by similar means DNA superstructure in nucleoids derived from a variety of different chick cells. Nucleoids derived from adult hen erythrocytes differ from the other nucleoids studied in that their sedimentation rate does not vary in the manner characteristic of supercoiled DNA.


Anti-tumor and anti-metastatic actions of wogonin isolated from Scutellaria baicalensis roots through anti-lymphangiogenesis.
Tumor growth and metastasis are associated with angiogenesis and lymphangiogenesis through the production of vascular endothelial growth factor (VEGF) or VEGF-C in tumors, and the phosphorylation of VEGF receptor (VEGFR)-2 or VEGFR-3 in vascular endothelial cells or lymphatic endothelial cells (LECs). Tumor-associated macrophages (TAMs) play an important role in tumor lymphangiogenesis, and consequently stimulate metastasis through the lymphatic system to lymph nodes. We examined the effects of wogonin isolated from Scutellaria baicalensis roots on tumor growth and metastasis using a highly metastatic model in osteosarcoma LM8-bearing mice. Wogonin (25 and 50 mg/kg, twice daily) reduced tumor growth and metastasis to the lung, liver and kidney, angiogenesis (CD31-positive cells), lymphangiogenesis (LYVE-1-positive cells), and TAM (F4/80-positive cell) numbers in the tumors of LM8-bearing mice. Wogonin (10-100 μM) also inhibited increases in IL-1β production and cyclooxygenase (COX)-2 expression induced by lipopolysaccharide in THP-1 macrophages. Wogonin had no effect on VEGF-C production in LM8 cells, or VEGFR-3 expression in human lymphatic endothelial cells (HLECs), however, it inhibited VEGF-C-induced VEGFR-3 phosphorylation in HLECs. The anti-tumor and anti-metastatic actions of wogonin may be associated with the inhibition of VEGF-C-induced lymphangiogenesis through a reduction in VEGF-C-induced VEGFR-3 phosphorylation by the inhibition of COX-2 expression and IL-1β production in TAMs.


[New clinical entity of myasthenia gravis with autoimmune targets of heart and skeletal muscles].
Since previous case reports have shown inflammatory myopathies in a few patients with myasthenia gravis (MG), heart and skeletal muscles are speculated to be autoimmune targets in MG. We screened to investigate the clinical, histological and immunological features of MG patients, who also developed myocarditis and/or myositis at 5 Japanese hospitals. Of 924 MG patients, eight (0.9%) had inflammatory myopathies. The onset age of MG was 55.3 ± 10.3 years. All patients showed severe symptoms with bulbar involvement accompanied by myasthenic crisis in 5 and invasive thymoma in 4. Myocarditis was found in 3 patients and myositis in 6. Myocarditis, developing 13-211 months after the MG onset, was characterized by heart failure and arrhythmias. Myositis, developing before or at the same time as MG, affected limb and paraspinal muscles. Histological findings of skeletal muscles showed CD8+lymphocyte infiltration. Seven patients had one of these anti-striational autoantibodies (those to titin, ryanodine receptor, muscular voltage-gated potassium channel, Kv1.4). Immunomodulatory therapy was required in all patients and was effective for both MG and inflammatory myopathies, except for one autopsy case. Heart and skeletal muscles are autoimmune targets in some MG patients. This autoimmunity has a broad clinical spectrum with anti-striational autoantibodies.


Beyond Intracellular Accumulation of Polyhydroxyalkanoates: Chiral Hydroxyalkanoic Acids and Polymer Secretion.
Polyhydroxyalkanoates (PHAs) are ubiquitous prokaryotic storage compounds of carbon and energy, acting as sinks for reducing power during periods of surplus of carbon source relative to other nutrients. With close to 150 different hydroxyalkanoate monomers identified, the structure and properties of these polyesters can be adjusted to serve applications ranging from food packaging to biomedical uses. Despite its versatility and the intensive research in the area over the last three decades, the market share of PHAs is still low. While considerable rich literature has accumulated concerning biochemical, physiological, and genetic aspects of PHAs intracellular accumulation, the costs of substrates and processing costs, including the extraction of the polymer accumulated in intracellular granules, still hampers a more widespread use of this family of polymers. This review presents a comprehensive survey and critical analysis of the process engineering and metabolic engineering strategies reported in literature aimed at the production of chiral (R)-hydroxycarboxylic acids (RHAs), either from the accumulated polymer or by bypassing the accumulation of PHAs using metabolically engineered bacteria, and the strategies developed to recover the accumulated polymer without using conventional downstream separations processes. Each of these topics, that have received less attention compared to PHAs accumulation, could potentially improve the economy of PHAs production and use. (R)-hydroxycarboxylic acids can be used as chiral precursors, thanks to its easily modifiable functional groups, and can be either produced de-novo or be obtained from recycled PHA products. On the other hand, efficient mechanisms of PHAs release from bacterial cells, including controlled cell lysis and PHA excretion, could reduce downstream costs and simplify the polymer recovery process.


A retrospective comparison of the influence of surgical clipping and endovascular embolization on recovery of oculomotor nerve palsy in patients with posterior communicating artery aneurysms.
Oculomotor nerve palsy (ONP) is a common symptom of posterior communicating artery aneurysms (PcomAAs). Surgical clipping and endovascular embolization are used to treat PcomAAs with ONP. To analyze the impact of these 2 techniques on recovery of ONP caused by PcomAAs. The clinical data for 176 patients with intracranial PcomAAs with ONP admitted to the Department of Neurosurgery, Sichuan Provincial People's Hospital, between June 2008 and May 2013 who undergone surgical clipping or endovascular embolization were studied retrospectively. The 2 treatment groups were compared with respect to age, sex, aneurysm size, levels of hypertension and hyperlipidemia, preadmission ONP duration, subarachnoid hemorrhage (SAH), complete ONP, postoperative recovery time from ONP symptoms, and degree of recovery. The follow-up duration was a minimum of 12 months. Multivariate Cox regression was used for analysis. A total of 132 patients were treated by surgical clipping, and 44 were treated by endovascular embolization. Significant differences were found in postoperative recovery time (83.87 ± 34.70 days for clipping and 137.45 ± 44.94 days for embolization, P < .001) and recovery rates (130 [98.5%] for clipping and 30 [68.2%] for embolization, P < .001). The period between ONP onset and admission was associated with recovery. Postoperative complications included significant cerebral vasospasms (6 in the clipping group and 2 in the embolization group) and hydrocephalus (16 in the clipping group and 9 in the embolization group). Simultaneous elimination of 2 injury mechanisms, compression and pulsation, when treating the oculomotor nerve by surgical clipping may be more advantageous than endovascular embolization to treat ONP caused by PcomAA.


Videoscope-Assisted Minimally Invasive Surgery (VMIS) for Bone Regeneration around Teeth and Implants: A Literature Review and Technique Update.
Background—The literature related to minimally invasive periodontal surgery is reviewed. This includes the original minimally invasive surgery (MIS) procedure for bone regeneration, the modification of MIS for the minimally invasive surgery technique (MIST) and modified MIST (M-MIST) procedures, and the introduction of the videoscope for oral surgical procedures and the ability to perform videoscope-assisted minimally invasive surgery (VMIS). The evolution from MIS through MIST to the current VMIS is reviewed. The results from studies of each of these methods are reported. Conclusion—The use of small incisions that produce minimal trauma and preserve most of the blood supply to the periodontal and peri-implant tissues results in improved regenerative outcomes, minimal to absent negative esthetic outcomes, and little or no patient discomfort. Minimally invasive procedures are a reliable method to regenerate periodontal tissues.


Meiotic spindle normality predicts live birth in patients with recurrent in vitro fertilization failure.
To investigate the impact of meiotic spindle normality on live birth rates in women with recurrent IVF failure. A retrospective comparative study. A large private sector IVF service in Sydney, Australia. Five hundred five intracytoplasmic sperm injection (ICSI)/ET cycles of patients who fulfilled the criteria for recurrent IVF failure, three or more previous fresh or frozen ETs with no ongoing pregnancy. Polarized light microscopy was used at the time of ICSI. Clinical pregnancy rates (PR) and live birth rates (LBR) were compared depending on the morphology of the meiotic spindle of the oocyte(s) from which the embryo(s) were transferred. Women receiving embryos where at least one was derived from a normally spindled oocyte had significantly increased clinical PR and LBR when compared with those who had only embryos derived from abnormally spindled oocytes (clinical PR: 31% vs. 7%; odds ratio [OR], 6.45; 95% confidence interval [CI] 3.65-11.40) and (LBR: 24% vs. 4%; OR, 7.24; 95% CI 3.62-14.49). Comparison between the abnormal spindle groups showed significantly higher clinical PR and LBR from the group of patients receiving embryos where at least one was derived from an oocyte with no visible spindle compared with the group receiving embryos from dysmorphic spindles only (clinical PR: 9% vs. 6%; OR, 0.58; 95% CI 0.22-1.57 and LBR: 8% vs. 1%; OR, 0.16; 95% CI 0.03-0.77). Normally spindled oocytes, as determined by polarized light microscopy, are associated with significantly higher clinical PRs in patients with recurrent IVF failure.


Tools for the Microbiome: Nano and Beyond.
The microbiome presents great opportunities for understanding and improving the world around us and elucidating the interactions that compose it. The microbiome also poses tremendous challenges for mapping and manipulating the entangled networks of interactions among myriad diverse organisms. Here, we describe the opportunities, technical needs, and potential approaches to address these challenges, based on recent and upcoming advances in measurement and control at the nanoscale and beyond. These technical needs will provide the basis for advancing the largely descriptive studies of the microbiome to the theoretical and mechanistic understandings that will underpin the discipline of microbiome engineering. We anticipate that the new tools and methods developed will also be more broadly useful in environmental monitoring, medicine, forensics, and other areas.


Costs of Joint Replacement in Osteoarthritis: A Study Using the National Joint Registry and Clinical Practice Research Datalink Data Sets.
To estimate the costs of primary hip and knee replacement in individuals with osteoarthritis up to 2 years postsurgery, compare costs before and after the surgery, and identify predictors of hospital costs. Patients age ≥18 years with primary planned hip or knee replacements and osteoarthritis in England between 2008 and 2016 were identified from the National Joint Registry and linked with Hospital Episode Statistics data containing inpatient episodes. Primary care data linked with hospital outpatient records were also used to identify patients age ≥18 years with primary hip or knee replacements between 2008 and 2016. All health care resource use was valued using 2016/2017 costs, and nonparametric censoring methods were used to estimate total 1-year and 2-year costs. We identified 854,866 individuals undergoing hip or knee replacement. The mean censor-adjusted 1-year hospitalization costs for hip and knee replacement were £7,827 (95% confidence interval [95% CI] 7,813, 7,842) and £7,805 (95% CI 7,790, 7,818), respectively. Complications and revisions were associated with up to a 3-fold increase in 1-year hospitalization costs. The censor-adjusted 2-year costs were £9,258 (95% CI 9,233, 9,280) and £9,452 (95% CI 9,430, 9,475) for hip and knee replacement, respectively. Adding primary and outpatient care, the mean total hip and knee replacement 2-year costs were £11,987 and £12,578, respectively. There are significant costs following joint replacement. Revisions and complications accounted for considerable costs and there is a significant incentive to identify best approaches to reduce these.


Organ transplantation in Greece: the need for mediation.
Organ transplants are not regarded as an exclusively medical process, because they involve financial, religious, philosophic, and bioethical parameters. It becomes clear that if they are to achieve their purpose, which we believe extends well beyond the medical dimension, the creation of a comprehensive framework of communication between the involved parties is of paramount importance. The aim of this paper is to present an outline and a number of considerations regarding the communicational, bioethical, and legal issues that arise from a rather dramatic state of affairs in Greece today: In 2012 the rate of organ transplants stood at only 7 per 1 million of the population. The outdated legal framework and the lack of trust on the part of patients and the public have led to a highly inefficient system that is lagging behind in many respects. The proposition made in this paper is that there is a need for a new system of communication between doctors, patients, relatives of patients, and hospitals: bioethical mediation. This is a system that has played a vital role and has produced astounding results in other countries. There is also every indication that the introduction of such a system is crucial for Greece, especially as the symptoms of the acute financial crisis are become fully visible and tangible. Mediation aims to identify solutions that are oriented toward the interests and wishes of patients, are acknowledged and accepted by all parties involved, and are in tune with the values and the principles of medical practice.


Management of a pet dog after exposure to a human patient with Ebola virus disease.
In October 2014, a health-care worker who had been part of the treatment team for the first laboratory-confirmed case of Ebola virus disease imported to the United States developed symptoms of Ebola virus disease. A presumptive positive reverse transcription PCR assay result for Ebola virus RNA in a blood sample from the worker was confirmed by the CDC, making this the first documented occurrence of domestic transmission of Ebola virus in the United States. The Texas Department of State Health Services commissioner issued a control order requiring disinfection and decontamination of the health-care worker's residence. This process was delayed until the patient's pet dog (which, having been exposed to a human with Ebola virus disease, potentially posed a public health risk) was removed from the residence. This report describes the movement, quarantine, care, testing, and release of the pet dog, highlighting the interdisciplinary, one-health approach and extensive collaboration and communication across local, county, state, and federal agencies involved in the response.


The conformation of the pore region of the M2 proton channel depends on lipid bilayer environment.
The M2 protein from influenza A virus is a 97-amino-acid protein with a single transmembrane helix that forms proton-selective channels essential to virus function. The hydrophobic transmembrane domain of the M2 protein (M2TM) contains a sequence motif that mediates the formation of functional tetramers in membrane environments. A variety of structural models have previously been proposed which differ in the degree of helix tilt, with proposed tilts ranging from approximately 15 degrees to 38 degrees . An important issue for understanding the structure of M2TM is the role of peptide-lipid interactions in the stabilization of the lipid bilayer bound tetramer. Here, we labeled the N terminus of M2TM with a nitroxide and studied the tetramer reconstituted into lipid bilayers of different thicknesses using EPR spectroscopy. Analyses of spectral changes provide evidence that the lipid bilayer does influence the conformation. The structural plasticity displayed by M2TM in response to membrane composition may be indicative of functional requirements for conformational change. The various structural models for M2TM proposed to date--each defined by a different set of criteria and in a different environment--might provide snapshots of the distinct conformational states sampled by the protein.


Survey of Schistosomiasis in Saint Lucia: Evidence for Interruption of Transmission.
Saint Lucia at one time had levels of schistosomiasis prevalence and morbidity as high as many countries in Africa. However, as a result of control efforts and economic development, including more widespread access to sanitation and safe water, schistosomiasis on the island has practically disappeared. To evaluate the current status of schistosomiasis in Saint Lucia, we conducted a nationally representative school-based survey of 8-11-year-old children for prevalence of Schistosoma mansoni infections using circulating antigen and specific antibody detection methods. We also conducted a questionnaire about available water sources, sanitation, and contact with fresh water. The total population of 8-11-year-old children on Saint Lucia was 8,985; of these, 1,487 (16.5%) provided urine for antigen testing, 1,455 (16.2%) provided fingerstick blood for antibody testing, and 1,536 (17.1%) answered the questionnaire. Although a few children were initially low positives by antigen or antibody detection methods, none could be confirmed positive by follow-up testing. Most children reported access to clean water and sanitary facilities in or near their homes and 48% of the children reported contact with fresh water. Together, these data suggest that schistosomiasis transmission has been interrupted on Saint Lucia. Additional surveys of adults, snails, and a repeat survey among school-age children will be necessary to verify these findings. However, in the same way that research on Saint Lucia generated the data leading to use of mass drug administration for schistosomiasis control, the island may also provide the information needed for guidelines to verify interruption of schistosomiasis transmission.


The Sydney West Knowledge Portal: Evaluating the Growth of a Knowledge Portal to Support Translational Research.
The Sydney West Translational Cancer Research Centre is an organization funded to build capacity for translational research in cancer. Translational research is essential for ensuring the integration of best available evidence into practice and for improving patient outcomes. However, there is a low level of awareness regarding what it is and how to conduct it optimally. One solution to addressing this gap is the design and deployment of web-based knowledge portals to disseminate new knowledge and engage with and connect dispersed networks of researchers. A knowledge portal is an web-based platform for increasing knowledge dissemination and management in a specialized area. To measure the design and growth of an web-based knowledge portal for increasing individual awareness of translational research and to build organizational capacity for the delivery of translational research projects in cancer. An adaptive methodology was used to capture the design and growth of an web-based knowledge portal in cancer. This involved stakeholder consultations to inform initial design of the portal. Once the portal was live, site analytics were reviewed to evaluate member usage of the portal and to measure growth in membership. Knowledge portal membership grew consistently for the first 18 months after deployment, before leveling out. Analysis of site metrics revealed members were most likely to visit portal pages with community-generated content, particularly pages with a focus on translational research. This was closely followed by pages that disseminated educational material about translational research. Preliminary data from this study suggest that knowledge portals may be beneficial tools for translating new evidence and fostering an environment of communication and collaboration.


The Role of the IL-20 Subfamily in Glaucoma.
Glaucoma is a common disease that leads to loss of peripheral vision and, if left untreated, ultimately to blindness. While the exact cause(s) of glaucoma is still unknown, two leading risk factors are age and elevated intraocular pressure. Several studies suggest a possible link between glaucoma and inflammation in humans and animal models. In particular, our lab recently identified a T104M mutation in IL-20 receptor-B (IL-20RB) in primary open angle glaucoma patients from a large pedigree. Several of the interleukin- (IL-) 20 family of cytokines and receptors are expressed in ocular tissues including the trabecular meshwork, optic nerve head, and retinal ganglion cells. The DBA/2J mouse develops high intraocular pressures with age and has characteristic optic nerve defects that make it a useful glaucoma model. IL-24 expression is significantly upregulated in the retina of these mice, while IL-20RA expression in the optic nerve is downregulated following pressure-induced damage. The identification of a mutation in the IL-20RB gene in a glaucoma pedigree and changes in expression levels of IL-20 family members in the DBA/2J mouse suggest that disruption of normal IL-20 signaling in the eye may contribute to degenerative processes associated with glaucoma.


Hydroxychloroquine and azithromycin as a treatment of COVID-19: results of an open-label non-randomized clinical trial.
Chloroquine and hydroxychloroquine have been found to be efficient on SARS-CoV-2, and reported to be efficient in Chinese COV-19 patients. We evaluate the effect of hydroxychloroquine on respiratory viral loads. French Confirmed COVID-19 patients were included in a single arm protocol from early March to March 16th, to receive 600mg of hydroxychloroquine daily and their viral load in nasopharyngeal swabs was tested daily in a hospital setting. Depending on their clinical presentation, azithromycin was added to the treatment. Untreated patients from another center and cases refusing the protocol were included as negative controls. Presence and absence of virus at Day6-post inclusion was considered the end point. Six patients were asymptomatic, 22 had upper respiratory tract infection symptoms and eight had lower respiratory tract infection symptoms. Twenty cases were treated in this study and showed a significant reduction of the viral carriage at D6-post inclusion compared to controls, and much lower average carrying duration than reported in the litterature for untreated patients. Azithromycin added to hydroxychloroquine was significantly more efficient for virus elimination. Despite its small sample size, our survey shows that hydroxychloroquine treatment is significantly associated with viral load reduction/disappearance in COVID-19 patients and its effect is reinforced by azithromycin.


The structure of the cyanobactin domain of unknown function from PatG in the patellamide gene cluster.
Patellamides are members of the cyanobactin family of ribosomally synthesized and post-translationally modified cyclic peptide natural products, many of which, including some patellamides, are biologically active. A detailed mechanistic understanding of the biosynthetic pathway would enable the construction of a biotechnological `toolkit' to make novel analogues of patellamides that are not found in nature. All but two of the protein domains involved in patellamide biosynthesis have been characterized. The two domains of unknown function (DUFs) are homologous to each other and are found at the C-termini of the multi-domain proteins PatA and PatG. The domain sequence is found in all cyanobactin-biosynthetic pathways characterized to date, implying a functional role in cyanobactin biosynthesis. Here, the crystal structure of the PatG DUF domain is reported and its binding interactions with plausible substrates are investigated.


Accelerated re-entrainment to advanced light cycles in BALB/cJ mice.
Circadian rhythms in mammals are coordinated by the suprachiasmatic nuclei (SCN) of the hypothalamus, which are most potently synchronized to environmental light-dark cycles. Large advances in the light-dark cycle typically yield gradual advances in activity rhythms on the order of 1-2h per day until re-entrainment is complete due to limitations on the circadian system which are not yet understood. In humans, this delay until re-entrainment is accomplished is experienced as jetlag, with accompanying symptoms of malaise, decreased cognitive performance, sleep problems and gastrointestinal distress. In these experiments, locomotor rhythms of BALB/cJ mice monitored by running wheels were shown to re-entrain to large 6- or 8-hour shifts of the light-dark cycle within 1-2 days, as opposed to the 5-7 days required for C57BL/6J mice. A single-day 6-hour advance of the LD cycle followed by release to constant darkness yielded similar phase shifts, demonstrating that exaggerated re-entrainment is not explained by masking of activity by the light-dark cycle. Responses in BALB/cJ mice were similar when monitored instead by motion detectors, indicating that wheel-running exercise does not influence the magnitude of responses. Neither brief (15 min) light exposure late during subjective nighttime nor 6-hour delays of the light-dark cycle produced exaggerated locomotor phase shifts, indicating that BALB/cJ mice do not merely experience enhanced sensitivity to light. Fos protein was expressed in cells of the SCN following acute light exposure at ZT10 of their previous light-dark cycle, a normally non-responsive time in the circadian cycle, but only in BALB/cJ (and not C57BL/6J) mice that had been subjected two days earlier to a single-day 6-hour advance of the light-dark cycle, indicating that their SCN had been advanced by that treatment. BALB/cJ mice may thus serve as a useful comparative model for studying molecular and physiological processes that limit responsiveness of circadian clocks to photic input.


Cancer, physical activity, and exercise.
This review examines the relationship between physical activity and cancer along the cancer continuum, and serves as a synthesis of systematic and meta-analytic reviews conducted to date. There exists a large body of epidemiologic evidence that conclude those who participate in higher levels of physical activity have a reduced likelihood of developing a variety of cancers compared to those who engage in lower levels of physical activity. Despite this observational evidence, the causal pathway underlying the association between participation in physical activity and cancer risk reduction remains unclear. Physical activity is also a useful adjunct to improve the deleterious sequelae experienced during cancer treatment. These deleterious sequelae may include fatigue, muscular weakness, deteriorated functional capacity, and many others. The benefits of physical activity during cancer treatment are similar to those experienced after treatment. Despite the growing volume of literature examining physical activity and cancer across the cancer continuum, a number of research gaps exist. There is little evidence on the safety of physical activity among all cancer survivors, as most trials have selectively recruited participants. The specific dose of exercise needed to optimize primary cancer prevention or symptom control during and after cancer treatment remains to be elucidated.


Calpain hydrolysis of alpha- and beta2-adaptins decreases clathrin-dependent endocytosis and may promote neurodegeneration.
Clathrin-dependent endocytosis is mediated by a tightly regulated network of molecular interactions that provides essential protein-protein and protein-lipid binding activities. Here we report the hydrolysis of the alpha- and beta2-subunits of the tetrameric adaptor protein complex 2 by calpain. Calcium-dependent alpha- and beta2-adaptin hydrolysis was observed in several rat tissues, including brain and primary neuronal cultures. Neuronal alpha- and beta2-adaptin cleavage was inducible by glutamate stimulation and was accompanied by the decreased endocytosis of transferrin. Heterologous expression of truncated forms of the beta2-adaptin subunit significantly decreased the membrane recruitment of clathrin and inhibited clathrin-mediated receptor endocytosis. Moreover, the presence of truncated beta2-adaptin sensitized neurons to glutamate receptor-mediated excitotoxicity. Proteolysis of alpha- and beta2-adaptins, as well as the accessory clathrin adaptors epsin 1, adaptor protein 180, and the clathrin assembly lymphoid myeloid leukemia protein, was detected in brain tissues after experimentally induced ischemia and in cases of human Alzheimer disease. The present study further clarifies the central role of calpain in regulating clathrin-dependent endocytosis and provides evidence for a novel mechanism through which calpain activation may promote neurodegeneration: the sensitization of cells to glutamate-mediated excitotoxicity via the decreased internalization of surface receptors.


Management of Open Pneumothorax in Tactical Combat Casualty Care: TCCC Guidelines Change 13-02.
During the recent United States Central Command (USCENTCOM) and Joint Trauma System (JTS) assessment of prehospital trauma care in Afghanistan, the deployed director of the Joint Theater Trauma System (JTTS), CAPT Donald R. Bennett, questioned why TCCC recommends treating a nonlethal injury (open pneumothorax) with an intervention (a nonvented chest seal) that could produce a lethal condition (tension pneumothorax). New research from the U.S. Army Institute of Surgical Research (USAISR) has found that, in a model of open pneumothorax treated with a chest seal in which increments of air were added to the pleural space to simulate an air leak from an injured lung, use of a vented chest seal prevented the subsequent development of a tension pneumothorax, whereas use of a nonvented chest seal did not. The updated TCCC Guideline for the battlefield management of open pneumothorax is: ?All open and/ or sucking chest wounds should be treated by immediately applying a vented chest seal to cover the defect. If a vente chest seal is not available, use a non-vented chest seal. Monitor the casualty for the potential development of a subsequent tension pneumothorax. If the casualty develops increasing hypoxia, respiratory distress, or hypotension and a tension pneumothorax is suspected, treat by burping or removing the dressing or by needle decompression.? This recommendation was approved by the required two-thirds majority of the Committee on TCCC in June 2013.


Suppression of rat cytomegalovirus replication by antibodies against gamma interferon.
The role of gamma interferon (IFN-gamma) in the resolution of rat cytomegalovirus (RCMV) infection was investigated. In the spleen, IFN-gamma-producing cells reached maximum numbers on day 7 after infection. Prophylactic treatment with high doses of recombinant rat IFN-gamma exerted antiviral activity in fibroblasts and protected immunosuppressed rats against a lethal RCMV challenge. Remarkably, in immunocompetent rats, neutralization of endogenous IFN-gamma activity significantly reduced the numbers of RCMV antigen-expressing cells in the spleen, the predominant site of viral replication. Moreover, protection of radiation-immunosuppressed infected rats by transferred immune T cells was enhanced by coinjection of IFN-gamma neutralizing antibodies. The observations were paralleled by in vitro findings: low concentrations of IFN-gamma enhanced viral replication in both macrophages and fibroblasts. These data suggest that IFN-gamma can play different and even opposite roles in the regulation of RCMV replication in vivo; T lymphocytes may contribute to the progression of RCMV infection by secreting IFN-gamma.


Structural and mechanistic basis for preferential deadenylation of U6 snRNA by Usb1.
Post-transcriptional modification of snRNA is central to spliceosome function. Usb1 is an exoribonuclease that shortens the oligo-uridine tail of U6 snRNA, resulting in a terminal 2',3' cyclic phosphate group in most eukaryotes, including humans. Loss of function mutations in human Usb1 cause the rare disorder poikiloderma with neutropenia (PN), and result in U6 snRNAs with elongated 3' ends that are aberrantly adenylated. Here, we show that human Usb1 removes 3' adenosines with 20-fold greater efficiency than uridines, which explains the presence of adenylated U6 snRNAs in cells lacking Usb1. We determined three high-resolution co-crystal structures of Usb1: wild-type Usb1 bound to the substrate analog adenosine 5'-monophosphate, and an inactive mutant bound to RNAs with a 3' terminal adenosine and uridine. These structures, along with QM/MM MD simulations of the catalytic mechanism, illuminate the molecular basis for preferential deadenylation of U6 snRNA. The extent of Usb1 processing is influenced by the secondary structure of U6 snRNA.


Characteristics of tinnitus in a population of 555 patients: specificities of tinnitus induced by noise trauma.
Tinnitus is often associated with hearing loss of a known etiology. In this study, we compared tinnitus that appeared to be induced by noise trauma with that perceived to start in other circumstances in a population of 555 patients attending the specialist tinnitus clinic at the University Hospital in Montpellier, France. Patients had consulted for persistent tinnitus for 7 years from the onset of their symptoms. Among these tinnitus patients, 17% described their tinnitus as starting after excessive noise exposure. The patients who had a history of noise trauma had a symmetrical hearing loss, and no difference was seen in lateralization of tinnitus perception. This subset of patients was mainly male and on average was 10 years younger than other tinnitus patients. In this population, the hearing loss is significantly less than that measured in the other patients, even allowing for their younger age. Statistical analysis showed a significant correlation between a history of exposure to noise trauma and the presence of a high-pitched "whistling" tinnitus. The presence of whistling tinnitus was significantly correlated with high-frequency hearing loss. The intensity of tinnitus, measured using a visual analog scale, appeared to be stronger than the measured hearing loss would suggest.


Selection of highly specific and sensitive mRNA biomarkers for the identification of blood.
In the present work, we have evaluated eight reportedly blood-specific mRNA markers (HBB, HBA, ALAS2, CD3G, ANK1, PBGD, SPTB, AQP9) in an attempt to determine the most suitable ones for use in forensic applications based on their sensitivities, specificities and performance with casework samples. While varying levels of expression were observed, all markers were relatively sensitive requiring as little as 1ng of RNA input into the reverse transcription (RT) reaction. In singleplex reactions, seven of the eight analyzed blood markers (all except AQP9) demonstrated a high degree of specificity for blood. In multiplex reactions, non-reproducible cross-reactivity was observed for several of the mRNA markers, which was reduced and, in most cases, eliminated when less input total RNA was used. Additionally, some cross-reactivity was observed with tissue and animal samples. Despite differences in the observed sensitivity and specificity of the blood markers examined in this study, a number of the candidates appear to be suitable for inclusion in appropriately validated multiplex mRNA-based body fluid identification systems.


Synthesis and characterisation of first row transition metal complexes of functionalized 1,2,4-benzothiadiazines.
Reaction of the novel ligand 3-(2'-pyridyl)-benzo-1,2,4-thiadiazine (L) with the transition metal chloride salts MCl2·xH2O (M(II) = Mn, Fe, Co, Cu and Zn) in a 2 : 1 mole ratio afforded the mononuclear octahedral (high spin) complexes L2MCl2 (1a-1e respectively) in which L binds in a chelate fashion via N(2) and the pyridyl N atoms. In the case of CuCl2 the intermediate 1 : 1 four-coordinate complex LCuCl2 (2) was also isolated which adopts a polymeric structure with pseudo-square planar molecules linked via long Cu···S contacts (d(Cu···S) = 2.938(1) Å) in the apical position. In the presence of non-interacting ions, 3 : 1 complexes are isolated, exemplified by the reaction of L with Fe(CF3SO3)2 in a 3 : 1 ratio which affords the low spin complex [L3Fe][CF3SO3]2 (3). Reaction of L with VCl3 in a 2 : 1 mole ratio under aerobic conditions afforded the vanadyl complex [L2V(=O)Cl][Cl] (4).


Dynamic plasticity in coupled avian midbrain maps.
Internal mapping of the external environment is carried out using the receptive fields of topographic neurons in the brain, and in a normal barn owl the aural and visual subcortical maps are aligned from early experiences. However, instantaneous misalignment of the aural and visual stimuli has been observed to result in adaptive behavior, manifested by functional and anatomical changes of the auditory processing system. Using methods of information theory and statistical mechanics a model of the adaptive dynamics of the aural receptive field is presented and analyzed. The dynamics is determined by maximizing the mutual information between the neural output and the weighted sensory neural inputs, admixed with noise, subject to biophysical constraints. The reduced costs of neural rewiring, as in the case of young barn owls, reveal two qualitatively different types of receptive field adaptation depending on the magnitude of the audiovisual misalignment. By letting the misalignment increase with time, it is shown that the ability to adapt can be increased even when neural rewiring costs are high, in agreement with recent experimental reports of the increased plasticity of the auditory space map in adult barn owls due to incremental learning. Finally, a critical speed of misalignment is identified, demarcating the crossover from adaptive to nonadaptive behavior.


Maternal addiction, child maladjustment and socio-demographic risks: implications for parenting behaviors.
In this study we examined three parenting dimensions (involvement, autonomy, and limit-setting) and three potential determinants (maternal addiction, low SES and its correlates, and mothers' perceptions of their children's maladjustment) in order to disentangle features of parenting that are uniquely related to maternal addiction from those related to contextual determinants. We also examined conditional effects of low SES and its correlates on parenting. Based on a literature review and predictions arising from an ecological model of parenting, we expected that maternal addiction would be related with problems in parental involvement, but that the other parenting dimensions would be related with mothers' perceptions of children's maladjustment and low SES. Accordingly, we examined variance in each parenting dimensions accounted for by each of the three determinants, respectively. Subjects included 120 (69 opiate-addicted and 51 SES-matched comparison) mothers with children under 16 years of age. Children's maladaptive behavior was assessed with the Behavioral Assessment System for Children, and parental adjustment with the Parent Child Relationship Inventory. Direct effect predictions were confirmed and two conditional effects involving single status and family size were also found. Although many parenting problems have previously been attributed to maternal addiction, only parental involvement is directly related to being an addict; other parenting dimensions may be better explained by contextual factors.


A Randomized Controlled Study of a Novel Botanical Acne Spot Treatment.

OBJECTIVE: The study evaluated the tolerability and efficacy of a new presented treatment for acne. The product is an OTC topical gel consisting of 2% SA, which is also enriched in botanicals that have been shown to have anti-inflammatory properties.

DESIGN: The study was designed as a single-site, randomized, investigator-blinded, split-face 10-day study.

SETTING: Subjects enrolled with a minimum of 2 inflammatory papular acne lesions and 2 non-inflammatory open or closed comedones on both sides of the face in symmetrical locations, to the greatest degree possible. One side of each subject's face was randomly selected to receive the study treatment product.

PARTICIPANTS: 25 subjects, 15 female and 10 males, ages 12 to 43 years, suffering from mild to moderate acne.

Measurements: Study duration was 10 days, with study visits occurring at baseline (day 0), day 1, day 2, day 3, day 7, and day 10. Subjects underwent investigator facial evaluation and lesion assessment by dermatologist at each of the visit days. For the inflammatory lesions, the assessed parameters were erythema, elevation, induration, and overall impression. The assessed non-inflammatory parameters were elevation and overall impression.

Results: The observed difference between the treatment and the control group increased between day 1 and day 2 and reached an average of 15% to 20% with small varieties between the parameters and stayed similar across the remaining visits. Statistically significance (P less than 0.005) was achieved for all inflammatory and non-inflammatory tested parameters.

Conclusion: This study was performed to determine the safety, efficacy, and ease of use of a botanical acne treatment gel in providing a reduction in inflammatory acne lesion erythema, elevation, and induration. Erythema and elevation were the most influential parameters in inflammatory lesion with improvement noted after 2 days of application.

J Drugs Dermatol. 2017;16(6):599-603.
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Magnetic field assisted programming of particle shapes and patterns.
Anisotropic particles have generated an enormous amount of research interest due to their applications for drug delivery, electronic displays and as micromotors. However, up till now, there is no single protocol capable of generating particles of "patchy" composition with a variety of well-defined and predictable shapes. To address this, in this submission we dispersed magnetic nanoparticles (MNPs) in a non-magnetic fluid containing monomer and crosslinker. This solution was added to the surface of Teflon, which was submerged in the solvent 2,2,4-trimethylpentane. Under these conditions a round, stable droplet was formed on the Teflon. Upon exposure to a permanent magnet, the MNPs self-assembled into clusters with a variety shapes and sizes. The shape and size of the clusters depended on the magnetic field strength, which we controlled by systematically varying the distance between the magnet and the droplet. Interestingly, the shape of the liquid droplet was also influenced by the magnetic field. Upon polymerization, the MNP patterns and the droplet shape was preserved. We also show that very complex MNP patterns and particle shapes could be generated by controlling the distance between the drop and both a magnet above and below the droplet. In this case, the resulting patterns depended on whether the magnets were attracting or repelling each other, which was capable of changing the field lines that the MNPs align with. Overall, this approach is capable of generating particles with predictable MNP patterns and particle shapes without the use of any templates or complex synthetic steps. Furthermore, by using a sprayer (or similar approaches, e.g., ink jet printing) this technique can be easily scaled up to produce many complex anisotropic particles in a short amount of time.


Coronary artery surgery: the use of decision analysis.
The choice between coronary by-pass surgery and medical therapy in patients with angiographically documented coronary artery disease was examined. This decision analysis included consideration of patient preferences, severity of disease, prognosis with medical therapy, surgical mortality rate, graft patency rate, the probability that surgery will provide both short-term and long-term pain relief, and the probability that surgery will alter long-term survival. Coronary surgery was found to be the preferred therapy in many patients with disabling angina; it was rarely found to be the preferred therapy in asymptomatic patients, even those with proximal obstruction of the left anterior descending coronary artery. The therapeutic decision was strongly affected by differences in patient attitudes and differences in the past results of the prospective surgeon. This paper presents both a data base and a methodology that allow the physician to apply decision analysis to individual patients with coronary artery disease.


AlphaLISA for the determination of median levels of the free β subunit of human chorionic gonadotropin in the serum of pregnant women.
Measurement of the free β subunit of human chorionic gonadotropin (free β-hCG) in serum is useful for prenatal screening. Concentrations of free β-hCG vary in different races. Conventional assays used for such measurements have limitations. We applied the AlphaLISA to measure levels of free β-hCG in maternal serum during 8-20 weeks of gestation in women from southern China. Two anti-free β-hCG antibodies were used: one was coated on AlphaLISA acceptor beads and the one was biotinylated. The assay also contained donor beads coated with streptavidin. The AlphaLISA assay detection limit was 0.11 ng/mL, and the analytical range was 0.11-200 ng/mL. The intra- and interassay coefficients of variation were 1.32%-2.50% and 3.44%-5.45%, respectively. The correlation with commercial Eu(3+)-labeled free β-HCG-TRFIA assay was good (y = 1.045x + 1.580, r(2) = 0.978). Median levels of free β-hCG in maternal serum at 8-20 weeks gestation were higher in women from southern China compared with those reported in women from other countries. The AlphaLISA for free β-HCG could become the assay of choice for applications in clinical diagnostics. The established median value of free β-HCG is helpful in clinical diagnosis specific for southern Chinese women.


Arthroscopic Capsulolabral Reconstruction for Posterior Shoulder Instability in Patients 18 Years Old or Younger.
The purpose of this study was to determine clinical outcomes for pain, function, instability, and return to activity level and sport in patients 18 years old or younger, treated with arthroscopic capsulolabral reconstruction for posterior instability of the shoulder. We retrospectively reviewed 22 athletes (25 shoulders) with unidirectional recurrent posterior shoulder instability treated with arthroscopic posterior capsulolabral reconstruction from 2002 to 2009. The study group included 19 males and 3 females with a mean age of 17 years. Patients were evaluated at a mean of 63 months postoperatively with American Shoulder and Elbow Surgeons (ASES) composite scores and subset scores for pain, stability, and function, as well as Marx activity scores. Statistical analysis was performed for continuous and categorical variables with significance set at α=0.05. The overall mean postoperative ASES and Marx scores were 74.3 (SD±20) and 14.8 (SD±3.2), respectively. Twenty-three shoulders were stable at the time of final follow-up (92%). Two shoulders had traumatic recurrent episodes of posterior instability. Return to sport at the same level was achieved in 67% of athletes. Overall postoperative ASES scores were significantly higher in male patients (P=0.04), those with traumatic injuries (P=0.03), and in contact athletes (P<0.01). Postoperative Marx scores were significantly higher in male patients (P<0.01). Preoperative and postoperative range of motion were assessed and without significant difference. Arthroscopic capsulolabral reconstruction is an effective treatment for symptomatic unidirectional posterior glenohumeral instability in 18 years old or younger. In distinction to treatment of anterior instability, outcomes in this series were improved in males, contact athletes, and patients with a traumatic etiology of posterior glenohumeral instability. Level IV.


Quantifying the Impact of Seasonal and Short-term Manure Application Decisions on Phosphorus Loss in Surface Runoff.
Agricultural phosphorus (P) management is a research and policy issue due to P loss from fields and water quality degradation. Better information is needed on the risk of P loss from dairy manure applied in winter or when runoff is imminent. We used the SurPhos computer model and 108 site-years of weather and runoff data to assess the impact of these two practices on dissolved P loss. Model results showed that winter manure application can increase P loss by 2.5 to 3.6 times compared with non-winter applications, with the amount increasing as the average runoff from a field increases. Increased P loss is true for manure applied any time from late November through early March, with a maximum P loss from application in late January and early February. Shifting manure application to fields with less runoff can reduce P loss by 3.4 to 7.5 times. Delaying manure application when runoff is imminent can reduce P loss any time of the year, and sometimes quite significantly, but the number of times that application delays will reduce P loss is limited to only 3 to 9% of possible spreading days, and average P loss may be reduced by only 15% for winter-applied manure and 6% for non-winter-applied manure. Overall, long-term strategies of shifting manure applications to low runoff seasons and fields can potentially reduce dissolved P loss in runoff much more compared with near-term, tactical application decisions of avoiding manure application when runoff is imminent.


Early postnatal maternal trait anxiety is associated with the behavioural outcomes of children born preterm <33 weeks.
Maternal ante- and postnatal anxiety have been associated with children's socio-emotional development. Moreover, maternal anxiety has been studied as both a contributing factor and consequence of preterm birth, and children born preterm are more likely to develop behavioural problems compared to term-born controls. This study investigated the association between maternal anxiety measured soon after birth and mental health in 215 ex-preterm children, born at <33 weeks, who participated in the Evaluation of Preterm Imaging Study. Children were followed-up at a median age of 4.6 years (range 4.2-6.6), and received behavioural and cognitive evaluation. Maternal trait anxiety was assessed with the Spielberger State-Trait Anxiety Index at term corrected age. Primary outcome measures were children's Strengths and Difficulties Questionnaire (SDQ) and Social Responsiveness Scale 2 (SRS-2) scores, indicative of generalised psychopathology and autism symptomatology, respectively. IQ was assessed with the Wechsler Preschool and Primary Scales of Intelligence. The final sample, after excluding participants with missing data and multiple pregnancy (n = 75), consisted of 140 children (51.4% male). Results showed that increased maternal trait anxiety at term corrected age was associated with children's higher SDQ scores (β = 0.25, 95% CI 0.09-0.41, p = 0.003, f2 = 0.08) and SRS-2 scores (β = 0.15, 95% CI 0.02-0.28, p = 0.03, f2 = 0.04). Our findings indicate that children born preterm whose mothers are more anxious in the early postnatal period may show poorer mental health outcomes at pre-school age. Further research is needed to investigate preventative measures that can be offered to high-risk premature babies and their families.


HIV treatment produces economic returns through increased work and education, and warrants continued US support.
Federal expenditures are under scrutiny in the United States, and the merits of continuing and expanding the President's Emergency Plan for AIDS Relief (PEPFAR) to support access to antiretroviral therapy have become a topic of debate. A growing body of research on the economic benefits of treatment with antiretroviral therapy has important implications for these discussions. For example, research conducted since the inception of PEPFAR shows that HIV-infected adults who receive antiretroviral therapy often begin or resume productive work, and that children living in households with infected adults who are on treatment are more likely to attend school than those in households with untreated adults. These benefits should be considered when weighing the overall benefits of providing antiretroviral therapy against its costs, particularly in the context of discussions about the future of PEPFAR. A modest case can also be made in favor of having private companies in HIV-affected countries provide antiretroviral therapy to their employees and dependents, thus sharing some of the burden of funding HIV treatment.


Differential IKK/NF-κB Activity Is Mediated by TSC2 through mTORC1 in PTEN-Null Prostate Cancer and Tuberous Sclerosis Complex Tumor Cells.
The serine/threonine protein kinase Akt plays a critical role in regulating proliferation, growth, and survival through phosphorylation of different downstream substrates. The mTOR is a key target for Akt to promote tumorigenesis. It has been reported that Akt activates mTOR through phosphorylation and inhibition of the tuberous sclerosis complex (TSC) protein TSC2. Previously, it was demonstrated that mTOR activates IKK/NF-κB signaling by promoting IκB kinase (IKK) activity downstream of Akt in conditions deficient of PTEN. In this study, the mechanistic role of the tumor-suppressor TSC2 was investigated in the regulation of IKK/NF-κB activity in PTEN-null prostate cancer and in TSC2-mutated tumor cells. The results demonstrate that TSC2 inhibits IKK/NF-κB activity downstream of Akt and upstream of mTORC1 in a PTEN-deficient environment. However, TSC2 promotes IKK/NF-κB activity upstream of Akt and mTORC1 in TSC2 mutated tumor cells. These data indicate that TSC2 negatively or positively regulates IKK/NF-κB activity in a context-dependent manner depending on the genetic background. This study provides fundamental insight for understanding the molecular details by which TSC2/mTOR regulates NF-κB signaling in different tumors.