Gonadotropin stimulation of ovarian fractalkine expression and fractalkine augmentation of progesterone biosynthesis by luteinizing granulosa cells.
Recent studies indicated that ovarian functions are regulated by diverse paracrine factors induced by the preovulatory increases in circulating LH. Based on DNA microarray analyses and real-time RT-PCR, we found a major increase in the transcript levels of a chemokine fractalkine after human chorionic gonadotropin (hCG) treatment during the preovulatory period in gonadotropin-primed immature mice and rats. Although CX3CR1, the seven-transmembrane receptor for fractalkine, was also found in murine ovaries, its transcripts displayed minimal changes. Using tandem RT-PCR and immunohistochemistry, fractalkine transcripts and proteins were localized in cumulus, mural granulosa, and theca cells as well as the oocytes, whereas CX3CR1 was found in the same cells except the oocyte. Real-time RT-PCR further indicated the hCG induction of fractalkine transcripts in different ovarian compartments, with the highest increases found in granulosa cells. In cultured granulosa cells, treatment with fractalkine augmented hCG stimulation of progesterone but not estradiol and cAMP biosynthesis with concomitant increases in transcript levels for key steroidogenic enzymes (steroidogenic acute regulatory protein, CYP11A, and 3beta-hydroxysteroid dehydrogenase). In cultured preovulatory follicles, treatment with fractalkine also augmented progesterone production stimulated by hCG. Furthermore, treatment with fractalkine augmented the phosphorylation of P38 MAPK in cultured granulosa cells. The present data demonstrated that increases in preovulatory LH/hCG induce the expression of fractalkine to augment the luteinization of preovulatory granulosa cells and suggest the fractalkine/CX3CR1 signaling system plays a potential paracrine/autocrine role in preovulatory follicles.


Tumor-associated macrophages and macrophage-related immune checkpoint expression in sarcomas.
Early trials for immune checkpoint inhibitors in sarcomas have delivered mixed results, and efforts to improve outcomes now look to combinatorial strategies with novel immunotherapeutics, including some that target macrophages. To enhance our understanding of the sarcoma immune landscape, we quantified and characterized tumor-associated macrophage infiltration and expression of the targetable macrophage-related immune checkpoint CD47/SIRPα across sarcoma types. We surveyed immunohistochemical expression of CD68, CD163, CD47, and SIRPα in tissue microarrays of 1242 sarcoma specimens (spanning 24 types). Non-translocation sarcomas, particularly undifferentiated pleomorphic sarcoma and dedifferentiated liposarcoma, had significantly higher counts of both CD68+ and CD163+ macrophages than translocation-associated sarcomas. Across nearly all sarcoma types, macrophages outnumbered tumor-infiltrating lymphocytes and CD163+ (M2-like) macrophages outnumbered CD68+ (M1-like) macrophages. These findings were supported by data from The Cancer Genome Atlas, which showed a correlation between increasing macrophage contributions to immune infiltration and several measures of DNA damage. CD47 expression was bimodal, with most cases showing either 0% or >90% tumor cell staining, and the highest CD47 scores were observed in chordoma, angiosarcoma, and pleomorphic liposarcoma. SIRPα scores correlated well with CD47 expression. Given the predominance of macrophage infiltrates over tumor-infiltrating lymphocytes, the bias toward M2-like (immunosuppressive) macrophage polarization, and the generally high scores for CD47 and SIRPα, macrophage-focused immunomodulatory agents, such as CD47 or IDO-1 inhibitors, may be particularly worthwhile to pursue in sarcoma patients, alone or in combination with lymphocyte-focused agents.


[Tobacco prevention in apprentices--evaluation of a general practice project "not smoking is cool!"].
In recent years the incidence of adolescent smokers has increased steeply. Taking this into account, current anti-smoker campaigns should be reconsidered. Great importance must be attached to the apprentice group, since adolescents in professional training have frequently not been reached sufficiently by the preventive efforts. This study presents a tobacco prevention campaign among adolescents which was conducted in a large Swiss industrial company in 1998. A total of 430 apprentices took part in the prevention campaign which intentionally targeted adolescent behaviour. Of these, 40 apprentices were trained as mediators for tobacco prevention in advance. On the prevention day, the mediators communicated their know-how and experiences to their working groups. To evaluate the campaign, the apprentices were surveyed via standardised questionnaires on their smoking habits before the prevention campaign began. One month after the prevention day they were surveyed again on their current smoking habits and how the campaign had affected them. The study showed that apprentices smoke heavily despite high awareness of the negative consequences. Some 30% of the trainees admitted smoking regularly. Another 11% said they were occasional tobacco consumers. No significant gender difference was noticeable. Especially striking was that the adolescents often justified their own smoking as habit and dependence on tobacco. The prevention campaign was well received by the adolescents. It set about half of the smokers thinking over their smoking habits. No change in smoking habits, however, could be demonstrated one month after the campaign. Prevention among adolescents beyond the mandatory school period is a special challenge, since one cannot take advantage of the school's network. Within the scope of the campaign described here, it was possible to reach 430 apprentices of a major company. The study shows a great need for on-target smoking prevention in this sector. In the future, studies should examine whether tobacco prevention campaigns for apprentices should already contain secondary prevention elements from the outset. They should also assess if the effects could be further improved by introducing structural and other ongoing health-promotion measures.


Influence of electronic triage decision-support on hospital admission, left without being seen and time to physician initial assessment in the emergency department.
To explore the impact of the implementation of eCTAS, a real-time electronic decision-support tool, on hospital admission, rate of left without being seen, and time from triage to physician initial assessment. We conducted a cohort study using population-based administrative data from all Ontario emergency departments (EDs) that had implemented eCTAS for 9 months. We compared 6 months post-eCTAS data to the same 6 months the previous year (pre-eCTAS). We included triage encounters of adult (≥ 18 years) patients if they had one of 16 pre-specified, high-volume presenting complaints. Multivariable logistic regression and quantile regression models informed the effect of eCTAS on outcomes. We included data from 354,176 triage encounters from 31 EDs. There was a change in the distribution of triage scores post-eCTAS, with fewer patients classified as CTAS 2 and CTAS 3, and more patients classified as CTAS 1 and CTAS 4. Overall, hospital admission decreased post-eCTAS (adjusted OR: 0.98; 95% CI: 0.97 to 1.00), with fewer CTAS 2 and more CTAS 3 and CTAS 4 patients admitted post-eCTAS. The rate of left without being seen increased (2.8% vs. 3.0%; adjusted OR: 1.07; 95% CI: 1.03 to 1.11) post-eCTAS, while time to physician initial assessment proved similar pre and post-eCTAS. eCTAS implementation had little impact on admission, rate of left without being seen and time to physician initial assessment. eCTAS appears to reclassify patients from higher to lower acuity scores, resulting in higher admission rates for CTAS 3 and CTAS 4 patients. It remains unknown if this reclassification is appropriate.


Melatonin influences pancreatic cancerogenesis.
Pancreatic cancer has fatal prognosis because of the absence of early symptoms, late diagnosis and the resistance to radio- and chemotherapy. Melatonin, an indoleamine discovered in the pineal gland, has also been detected in the gastrointestinal system and its specific receptors have been identified in the pancreas. Some evidence indicates that melatonin could modulate the process of pancreatic oncogenesis: 1) Melatonin, as direct scavenger of radical oxygen and nitrogen species (ROS and RNS) and activator of antioxidant enzymes effectively protects the pancreatic tissue against oxidative stress and inflammatory damage. 2) In pancreatic carcinoma cell line (PANC-1) melatonin used at high doses affects the Bax/Bcl protein balance, and stimulates the expressions of caspase-9 and caspase-3, thus activating the mitochondrial pathway of apoptosis. On the contrary, low concentrations of melatonin turn on the production of anti-apoptotic heat shock proteins: HSP27, HSP70, and HSP90, which prevents the activation of caspase-3. 3) Melatonin reduces angiogenesis and decreases proliferation of endothelial cells through inhibition of vascular endothelial factor (VEGF). 4) Melatonin strengthens the immune defense of the organism via activation of peripheral effector T cells and suppression of T regulatory cells. 5) In animal studies melatonin has been found to increase the efficacy of oncostatic drugs, to reduce the side effects of chemotherapy and to decrease morbidity. These observations suggest that melatonin at high doses could be potentially taken into consideration as the supportive treatment in the therapy of pancreatic cancer, although the effect of melatonin on apoptosis requires further study.


Antiviral Activity of Carrageenans and Processing Implications.
Carrageenan and carrageenan oligosaccharides are red seaweed sulfated carbohydrates with well-known antiviral properties, mainly through the blocking of the viral attachment stage. They also exhibit other interesting biological properties and can be used to prepare different drug delivery systems for controlled administration. The most active forms are λ-, ι-, and κ-carrageenans, the degree and sulfation position being determined in their properties. They can be obtained from sustainable worldwide available resources and the influence of manufacturing on composition, structure, and antiviral properties should be considered. This review presents a survey of the antiviral properties of carrageenan in relation to the processing conditions, particularly those assisted by intensification technologies during the extraction stage, and discusses the possibility of further chemical modifications.


A Novel Approach to Clustering Genome Sequences Using Inter-nucleotide Covariance.
Classification of DNA sequences is an important issue in the bioinformatics study, yet most existing methods for phylogenetic analysis including Multiple Sequence Alignment (MSA) are time-consuming and computationally expensive. The alignment-free methods are popular nowadays, whereas the manual intervention in those methods usually decreases the accuracy. Also, the interactions among nucleotides are neglected in most methods. Here we propose a new Accumulated Natural Vector (ANV) method which represents each DNA sequence by a point in ℝ18. By calculating the Accumulated Indicator Functions of nucleotides, we can further find an Accumulated Natural Vector for each sequence. This new Accumulated Natural Vector not only can capture the distribution of each nucleotide, but also provide the covariance among nucleotides. Thus global comparison of DNA sequences or genomes can be done easily in ℝ18. The tests of ANV of datasets of different sizes and types have proved the accuracy and time-efficiency of the new proposed ANV method.


Annexin A11 is targeted by IgG4 and IgG1 autoantibodies in IgG4-related disease.
Immunoglobulin G4-related disease (IgG4-RD) is a multiorgan immune-mediated disease that predominantly affects the biliary tract (IgG4-associated cholangitis, IAC) and pancreas (autoimmune pancreatitis, AIP). We recently identified highly expanded IgG4+ B-cell receptor clones in blood and affected tissues of patients with IAC/AIP suggestive of specific (auto)antigenic stimuli involved in initiating and/or maintaining the inflammatory response. This study aimed to identify (auto)antigen(s) that are responsible for the clonal expansion of IgG4+ B cells in IgG4-RD. We screened sera of patients with IAC/AIP (n=50), in comparison to control sera of patients with primary sclerosing cholangitis (PSC) and pancreatobiliary malignancies (n=47), for reactivity against human H69 cholangiocyte lysates on immunoblot. Subsequently, target antigens were immunoprecipitated and analysed by mass spectrometry. Prominent reactivity against a 56 kDa protein was detected in human H69 cholangiocyte lysates exposed to sera of nine patients with IAC/AIP. Affinity purification and mass spectrometry analysis identified annexin A11, a calcium-dependent phospholipid-binding protein. Annexin A11-specific IgG4 and IgG1 antibodies were only detected in serum of patients with IgG4-RD of the biliary tract/pancreas/salivary glands and not in disease mimickers with PSC and pancreatobiliary malignancies. Epitope analysis showed that two annexin A11 epitopes targeted by IgG1 and IgG4 autoantibodies were shared between patients with IAC/AIP and IgG4 antibodies blocked binding of IgG1 antibodies to the shared annexin A11 epitopes. Our data suggest that IgG1-mediated pro-inflammatory autoreactivity against annexin A11 in patients with IgG4-RD may be attenuated by formation of annexin A11-specific IgG4 antibodies supporting an anti-inflammatory role of IgG4 in IgG4-RD.


Comprehensive Analysis of Expression Regulation for RNA m6A Regulators With Clinical Significance in Human Cancers.
N6-methyladenosine (m6A), the most abundant chemical modification on eukaryotic messenger RNA (mRNA), is modulated by three class of regulators namely "writers," "erasers," and "readers." Increasing studies have shown that aberrant expression of m6A regulators plays broad roles in tumorigenesis and progression. However, it is largely unknown regarding the expression regulation for RNA m6A regulators in human cancers. Here we characterized the expression profiles of RNA m6A regulators in 13 cancer types with The Cancer Genome Atlas (TCGA) data. We showed that METTL14, FTO, and ALKBH5 were down-regulated in most cancers, whereas YTHDF1 and IGF2BP3 were up-regulated in 12 cancer types except for thyroid carcinoma (THCA). Survival analysis further revealed that low expression of several m6A regulators displayed longer overall survival times. Then, we analyzed microRNA (miRNA)-regulated and DNA methylation-regulated expression changes of m6A regulators in pan-cancer. In total, we identified 158 miRNAs and 58 DNA methylation probes (DMPs) involved in expression regulation for RNA m6A regulators. Furthermore, we assessed the survival significance of those regulatory pairs. Among them, 10 miRNAs and 7 DMPs may promote cancer initiation and progression; conversely, 3 miRNA/mRNA pairs in kidney renal clear cell carcinoma (KIRC) may exert tumor-suppressor function. These findings are indicative of their potential prognostic values. Finally, we validated two of those miRNA/mRNA pairs (hsa-miR-1307-3p/METTL14 and hsa-miR-204-5p/IGF2BP3) that could serve a critical role for potential clinical application in KIRC patients. Our findings highlighted the importance of upstream regulation (miRNA and DNA methylation) governing m6A regulators' expression in pan-cancer. As a result, we identified several informative regulatory pairs for prognostic stratification. Thus, our study provides new insights into molecular mechanisms of m6A modification in human cancers.


The carcinogenicity of dietary acrylamide intake: a comparative discussion of epidemiological and experimental animal research.
Since 2002, it is known that the probable human carcinogen acrylamide is present in commonly consumed carbohydrate-rich foods, such as French fries and potato chips. In this review, the authors discuss the body of evidence on acrylamide carcinogenicity from both epidemiological and rodent studies, including variability, strengths and weaknesses, how both types of evidence relate, and possible reasons for discrepancies. In both rats and humans, increased incidences of various cancer types were observed. In rats, increased incidences of mammary gland, thyroid tumors and scrotal mesothelioma were observed in both studies that were performed. In humans, increased risks of ovarian and endometrial cancers, renal cell cancer, estrogen (and progesterone) receptor-positive breast cancer, and oral cavity cancer (the latter in non-smoking women) were observed. Some cancer types were found in both rats and humans, e.g., endometrial cancer (observed in one of the two rat studies), but there are also some inconsistencies. Interestingly, in humans, some indications for inverse associations were observed for lung and bladder cancers in women, and prostate and oro- and hypopharynx cancers in men. These latter observations indicate that genotoxicity may not be the only mechanism by which acrylamide causes cancer. The estimated risks based on the epidemiological studies for the sites for which a positive association was observed were considerably higher than those based on extrapolations from the rat studies. The observed pattern of increased risks in the rat and epidemiological studies and the decreased risks in the epidemiological studies suggests that acrylamide might influence hormonal systems, for which rodents may not be good models.


Potential therapeutic effects of vitamin e and C on placental oxidative stress induced by nicotine: an in vitro evidence.
There have been a few studies that examined the oxidative stress effects of nicotine during pregnancy and lactation. The adverse effect of prenatal smoking exposure on human fetal development and growth has been a major public health issue. Active or passive smoking during pregnancy can result in a wide variety of adverse outcomes, including intrauterine growth retardation (IUGR), prematurity, stillbirth, and the sudden infant death syndrome. Smoking in pregnancy has also been associated with an increased risk of attention deficit and learning problems in childhood. Some studies argued that as a principal component of tobacco smoke, nicotine alone is responsible for the majority of negative reproductive outcomes. Nicotine and its major metabolite cotinine can cross the placental barrier. The level of nicotine in fetal tissues was found to be equal to or greater than the plasma nicotine level in the mothers. The oxidative stress induce by nicotine has been increasingly postulated as a major contributor to endothelial dysfunction. A large body of research has investigated the potential role of antioxidant nutrients in the prevention of endothelial dysfunction in women. Therefore, the present study was undertaken to assess the potential benefit of antioxidant supplementation on markers of placental oxidative stress in an in vitro model of endothelial dysfunction induced by nicotine, since it was previously found that nicotine is able to trigger the placental secretion of stress molecules. In this regard, we evaluated the effects of vitamin C, vitamin E and N-acetylcysteine (NAC), alone or in combination, in placental villi culture after exposure to nicotine. The effect of antioxidant nutrients on trophoblast cells proliferation and vitality was also evaluated. The results obtained suggest that in a patho-physiological condition, such as endothelial dysfunction induced by nicotine, the deleterious effect of reactive oxygen species may be counteracted by an antioxidant therapy, and there is the need to investigate the optimum dosing and timing of antioxidants administration, since an inappropriate antioxidant treatment in pregnant women may have deleterious consequences, reducing placental cells proliferation until to cell death.


Phytophotodermatitis from Heracleum mantegazzianum.
Five individuals developed striate bullous dermatitis two days after exposure on a clear day to Heracleum mantegazzianum, Umbelliferae (Parsnip) family. Photopatch testing confirmed phototoxicity from the plant. This and 28 other Heracleum species yield linear furanocoumarins.


Urinary metabolic biomarkers of diet quality in European children are associated with metabolic health.
Urinary metabolic profiling is a promising powerful tool to reflect dietary intake and can help understand metabolic alterations in response to diet quality. Here, we used 1H NMR spectroscopy in a multicountry study in European children (1147 children from 6 different cohorts) and identified a common panel of 4 urinary metabolites (hippurate, N-methylnicotinic acid, urea, and sucrose) that was predictive of Mediterranean diet adherence (KIDMED) and ultra-processed food consumption and also had higher capacity in discriminating children's diet quality than that of established sociodemographic determinants. Further, we showed that the identified metabolite panel also reflected the associations of these diet quality indicators with C-peptide, a stable and accurate marker of insulin resistance and future risk of metabolic disease. This methodology enables objective assessment of dietary patterns in European child populations, complementary to traditional questionary methods, and can be used in future studies to evaluate diet quality. Moreover, this knowledge can provide mechanistic evidence of common biological pathways that characterize healthy and unhealthy dietary patterns, and diet-related molecular alterations that could associate to metabolic disease.


BPAG1n4 is essential for retrograde axonal transport in sensory neurons.
Disruption of the BPAG1 (bullous pemphigoid antigen 1) gene results in progressive deterioration in motor function and devastating sensory neurodegeneration in the null mice. We have previously demonstrated that BPAG1n1 and BPAG1n3 play important roles in organizing cytoskeletal networks in vivo. Here, we characterize functions of a novel BPAG1 neuronal isoform, BPAG1n4. Results obtained from yeast two-hybrid screening, blot overlay binding assays, and coimmunoprecipitations demonstrate that BPAG1n4 interacts directly with dynactin p150Glued through its unique ezrin/radixin/moesin domain. Studies using double immunofluorescent microscopy and ultrastructural analysis reveal physiological colocalization of BPAG1n4 with dynactin/dynein. Disruption of the interaction between BPAG1n4 and dynactin results in severe defects in retrograde axonal transport. We conclude that BPAG1n4 plays an essential role in retrograde axonal transport in sensory neurons. These findings might advance our understanding of pathogenesis of axonal degeneration and neuronal death.


Understanding the characteristics of friendship quality, activity participation, and emotional well-being in Taiwanese adolescents with autism spectrum disorder.
Autism spectrum disorder (ASD) is a lifelong developmental disability characterized by deficits in social communication and social interaction across multiple contexts. Existing literature on social relationships and well-being among adolescents with ASD in Asian countries is scant. This study compared the perceptions of adolescents with ASD with those of their neurotypical peers toward their friendship quality, activity participation, and emotional well-being, and examined the relationships between friendship quality, activity participation, and emotional well-being. The study participants-101 adolescents with ASD and 101 neurotypical peers, aged 10-19 years-completed the following self-administered questionnaires: the Friendship Quality Questionnaire, the Child and Adolescent Scale of Participation, the Beck Anxiety Inventory, and the short-form UCLA loneliness scale. Adolescents with ASD reported lower friendship quality, lower school participation, and higher levels of anxiety and loneliness compared to their neurotypical peers. Loneliness correlated negatively with friendship quality and school participation and positively with anxiety. Adolescents with ASD experienced increased levels of anxiety when low friendship quality was accompanied by greater loneliness. These findings reveal that friendship quality, school participation, and loneliness have a considerable effect on the emotional well-being of adolescents with ASD, thus indicating the need for therapeutic interventions that address interpersonal relationships and emotional well-being.


Maternal-fetal transmission of Toxoplasma gondii in interferon-gamma deficient pregnant mice.
Toxoplasma gondii infection is generally asymptomatic in immunocompetent persons but can be life-threatening in immunocompromised persons and for fetuses in the case of maternal-fetal transmission. The effect of interferon (IFN)-gamma, which plays a crucial role in the protective immunity against T. gondii infection, on maternal-fetal transmission of T. gondii was analyzed by quantitative competitive polymerase chain reaction targeting T. gondii-specific SAG1 gene. T. gondii loads were obvious in uterus and placenta of wild type (WT) C57BL/6 (B6, susceptible strain) but not BALB/c (resistant strain) pregnant mice. Higher levels of T. gondii were detected in uterus and placenta of IFN-gamma knock-out (GKO) B6 and BALB/c than in those of WT mice. Furthermore, T. gondii was detected in fetus of GKO B6 but not GKO BALB/c, WT B6, or WT BALB/c mice. Thus, not only IFN-gamma but also genetic susceptibility to T. gondii infection was important for the protective immunity of maternal-fetal transmission of T. gondii to fetus via placenta. T. gondii-infected WT mice displayed a low delivery rate with high IFN-gamma production, whereas infected GKO mice did not. Additionally, mean body weight of neonates from T. gondii-infected GKO BALB/c pregnant mice was significantly lower than that of unaborted neonates from WT BALB/c pregnant mice, suggesting the effects of T. gondii infection on intrauterine growth retardation of fetus in pregnant GKO mice.


The influence of pool volume and summer desiccation on the production of the resting and dispersal stage in a Daphnia metapopulation.
Dispersal is a key process in metapopulations, as migrants genetically connect populations and enable the colonization of empty habitat patches. Sub-populations may differ in their numerical contribution of migrants within a metapopulation. This has strong implications on evolutionary and ecological dynamics and has led to two different hypotheses about the Daphnia metapopulation studied here: the assessment by some authors is that sub-populations contribute equally to the production of migrants, while others have postulated long-lived core populations in large "mainland" habitat patches as the dominant source of migrants. We have studied the resting and dispersal stage (ephippium) in a natural Daphnia metapopulation and in mesocosm experiments, and tested for effects of habitat size and summer desiccation. We found that a 1,000-fold increase in rock pool volume resulted on average in only in a 2.8-fold increase in ephippium production. Mesocosm experiments confirmed these results: a 1,000-fold increase of the mesocosms' volume resulted in a 7.2-fold increase in ephippium production. Additionally, we showed that ephippium production did not depend on the initial population size. Thus, populations in small pools may contribute only marginal fewer potential migrants in the whole metapopulation than populations in large pools. In a second mesocosm experiment we found that summer desiccation, which is a typical occurrence in small pools, is not detrimental for the populations. Daphnia hatched out of ephippia that were produced earlier within the same season and built up viable populations again. The substantial production of ephippia by populations in small pools suggests that these populations might be important for both the dynamics and global stability of metapopulations.


Static Suspension of the Paralyzed Face Utilizing the Midfacial Corridor: Anatomic Evaluation and Surgical Technique.
Fascia lata and tendon grafts are frequently utilized to support the paralyzed midface and to extend muscular reach in McLaughin style, orthodromic temporalis transfers. The grafts are frequently placed in a deep subcutaneous positioning that can lead to the development of a, bowstring deformity in the cheek. This paper describes insertion of tendon grafts into the midfacial corridor collectively formed by the buccal, submasseteric and superficial temporal spaces. Over a seven-year period, all patients that underwent insertion of facia lata and tendon grafts in the midfacial corridor were included. Demographic information, perioperative variables and clinical outcomes were collected and analyzed. A total of 22 patients were included with a mean age of 64.3 years (33-86). There were multiple etiologies for the facial weakness including acoustic neuroma (9.1%), Bell's palsy (13.6%), facial nerve schwannoma (9.1%), temporal bone fracture (4.6%) and malignancy (22.7%). Midfacial corridor grafts were utilized in combination with nerve transfers (V-VII and XII-VII) in nine patients, McLaughin style temporalis transfers in 12 and as a standalone procedure in one individual. During the study period, no patients exhibited a tethering, or concave deformity in the midface. Additionally, no impingement, difficulties with mastication, parotitis or hematoma were encountered. One patient developed a postoperative infection, that was successfully managed. Placement of tendon or fascia grafts for static support or tunneling of an orthodromic temporalis transfer through the midfacial corridor can be performed rapidly while providing midfacial support and avoiding the creation of visible cutaneous deformities.


[Length of the spinal dural sac, sex differences and correlation with the length of the spinal cord and vertebral column in adults].
In 94 corpses (59 male and 35 female) of mature persons the length of the spinal dura mater sac has been studied. The average length of the sac is 621 +/- 3 mm. In men its average length is 636 +/- 4 mm, it makes 40 mm more in length than that in women (596 +/- 4 mm). The length of various parts in the dura mater sac is not the same: the cervical part makes 23% of the whole length, the thoracic--47%, the lumbar--23%, the sacral--7%. In men the cervical part of the sac in average is 6 mm longer than the lumbar part, and in women--quite the reverse, it is 7 mm shorter than the lumbar part. The sacral part of the sac in women is 3 mm longer that that in men. The sex differences noted are statistically significant. It is stated that the length of the spinal cord, its dura mater and the vertebral column are related as 1:1.5:1.7, the length of their cervical parts--as 1:1.5:1.4. the thoracic--as 1:1.3:1.3, the lumbar--as 1:2.4:3, the sacrococcygeal--as 1:1.4:4.9, respectively. During ontogenesis the greatest increase in the dura mater sac takes place in the cervical part as compared to the spinal cord and the vertebral column; in the thoracic part the intensity of their growth is equal: in the lumbar and in the sacrococcygeal part the increase of the vertebral column is the greatest.


Auscultation and echocardiographic findings in Bull Terriers with and without polycystic kidney disease.
To investigate a possible association between Bull Terrier polycystic kidney disease (BTPKD) and cardiac disease, to determine the prevalence of mitral valve disease (MVD) and left ventricular outflow tract obstruction (LVOTO) in the Australian Bull Terrier population, and to compare auscultation and echocardiography in detection of cardiac disease in Bull Terriers. Ninety-nine Bull Terriers, ranging in age from 8 weeks to 13 years and 11 months were auscultated and examined using renal ultrasonography; 86 were also examined using echocardiography. The prevalence and severity of heart defects in dogs with BTPKD was compared with that in dogs without BTPKD. Nineteen of these 99 dogs were diagnosed with BTPKD. Forty-two percent of Bull Terriers with BTPKD and 28% of those without BTPKD had murmurs characteristic of mitral regurgitation or LVOTO. How recently an animal was descended from an ancestor with BTPKD was associated with presence (P = 0.008) and loudness of a murmur (P = 0.009). Overall, echocardiography detected MVD in 39% of Bull Terriers, with increased prevalence in older animals (P = 0.003). Mitral stenosis was found in eight cases. Fifty-three percent of dogs in this study had evidence of LVOTO, with obstruction consisting of a complex of lesions including dynamic or fixed subvalvular LVOTO, significantly narrowed left ventricular outflow tract or valvular aortic stenosis. Dogs with BTPKD, or those descended from dogs with BTPKD, were more likely to have MVD (P = 0.006), and while LVOTO was not more common in these dogs, if they did have LVOTO, they were more likely to have severe obstruction than dogs with no ancestors with BTPKD (analysed in three ways P = 0.028 to 0.001). In this study, 46% of Bull Terriers without a murmur or arrhythmia had cardiac disease detected on echocardiographic examination. Cardiac disease, especially MVD and LVOTO, was common in Bull Terriers in this study, and those with BTPKD had an increased risk of cardiac abnormalities. Auscultation did not detect a significant number of Bull Terriers with cardiac disease.


Dysregulation of local stem/progenitor cells as a common cellular mechanism for heterotopic ossification.
Heterotopic ossification (HO), the abnormal formation of true marrow-containing bone within extraskeletal soft tissues, is a serious bony disorder that may be either acquired or hereditary. We utilized an animal model of the genetic disorder fibrodysplasia ossificans progressiva to examine the cellular mechanisms underlying HO. We found that HO in these animals was triggered by soft tissue injuries and that the effects were mediated by macrophages. Spreading of HO beyond the initial injury site was mediated by an abnormal adaptive immune system. These observations suggest that dysregulation of local stem/progenitor cells could be a common cellular mechanism for typical HO irrespective of the signal initiating the bone formation.


Syk-independent tyrosine phosphorylation and association of the protein tyrosine phosphatases SHP-1 and SHP-2 with the high affinity IgE receptor.
Aggregation of the high affinity IgE receptor (Fc epsilonRI), a member of the immune receptor family, results in the activation of protein tyrosine kinases and downstream signaling pathways. The two cytoplasmic Src homology 2 domain-containing protein tyrosine phosphatases, SHP-1 (also called SH-PTP1, PTP1C or HCP) and SHP-2 (also known as SH-PTP2, PTP1D, PTP2C, or Syp), are expressed in the RBL-2H3 rat mast cell line. Here we report that aggregation of Fc epsilonRI induced the tyrosine phosphorylation of both SHP-1 and SHP-2. This phosphorylation was independent of the presence of the protein tyrosine kinase Syk. Both SHP-1 and SHP-2 associated with Fc epsilonRI. Whereas SHP-1 was constitutively associated with the receptor, SHP-2 coprecipitated with Fc epsilonRI only after receptor aggregation. Fusion proteins containing either the full-length or the Src homology 2 domains of SHP-2 directly bound to the tyrosine-phosphorylated beta, but not the gamma, subunit of Fc epsilonRI. In the reciprocal experiments, synthetic phosphorylated peptides based on the immunoreceptor tyrosine-based activation motif of the beta, but not the gamma, subunit precipitated SHP-2. In contrast, neither fusion proteins nor synthetic peptides detected interaction between SHP-1 and Fc epsilonRI. In vitro, both SHP-1 and SHP-2 dephosphorylated tyrosine-phosphorylated beta and gamma subunits of Fc epsilonRI. Therefore, SHP-1 and SHP-2 associate with Fc epsilonRI by different mechanisms and can regulate the extent of the tyrosine phosphorylation of the receptor subunits. Thus, unlike other immune cells in which inhibitory molecules are recruited by accessory proteins, Fc epsilonRI bind molecules that both activate and inhibit signal transduction.


Lessons Learned From a Program to Reduce Diabetes Risk Among Low-Income Hispanic Women in a Community Health Clinic.
The Diabetes Prevention Program (DPP) and Look AHEAD studies demonstrated that modest weight loss and increased physical activity can significantly reduce the incidence of diabetes among overweight individuals with prediabetes. However, these studies involved costly interventions, all of which are beyond the reach of most real-world settings serving high-risk, low-income populations. Our project, De Por Vida, implemented a diabetes risk-reduction intervention for Hispanic women in a Federally Qualified Health Center and assessed the program's efficacy. This report describes the methodology used to develop and implement De Por Vida, the cultural adaptations made, the community-academic partnership formed to carry out this program, and the barriers and challenges encountered through the implementation process. Our goal was to translate the DPP and Look AHEAD programs into an intervention to prevent diabetes and reduce diabetes complications among high-risk Hispanic women at a federally qualified health center in Hillsboro, Oregon, where more than half of clinic patients are Spanish-speaking, and nearly all live in poverty. This randomized clinical trial targeted overweight Spanish-speaking women at risk for, or diagnosed with, type 2 diabetes. We developed a 12-month behavioral diabetes risk-reduction intervention that was responsive to the cultural practices of the Hispanic population and that could be implemented in low-income clinical settings. Study planning and implementation involved close collaboration among the clinic leadership, a research team from the Kaiser Permanente Center for Health Research, and Arizona State University. Creating a fully informed partnership between research and clinical institutions is the first step in successful cooperative research projects. The adoption of a bidirectional, rather than a top-down, approach to communication between researchers and health-care providers, and between clinic management and the clinic frontline staff, gave the research study team crucial information about barriers, constraints, and needs that clinic staff experienced in implementing the program. This allowed clinic management and front-line clinic staff to play an active role in study implementation, identifying problem areas, and collaborating in finding practical solutions. www.clinicaltrials.gov, NCT03113916.


Diverse nitrogen sources in seminal fluid act in synergy to induce filamentous growth of Candida albicans.
The pathogenic fungus Candida albicans is the leading cause of vulvovaginal candidiasis (VVC). VVC represents a major quality-of-life issue for women during their reproductive years, a stage of life where the vaginal epithelium is subject to periodic hormonally induced changes associated with menstruation and concomitant exposure to serum as well as potential intermittent contact with seminal fluid. Seminal fluid potently triggers Candida albicans to switch from yeastlike to filamentous modes of growth, a developmental response tightly linked to virulence. Conversely, vaginal fluid inhibits filamentation. Here, we used artificial formulations of seminal and vaginal fluids that faithfully mimic genuine fluids to assess the contribution of individual components within these fluids to filamentation. The high levels of albumin, amino acids, and N-acetylglucosamine in seminal fluid act synergistically as potent inducers of filamentous growth, even at atmospheric levels of CO2 and reduced temperatures (30 °C). Using a simplified in vitro model that mimics the natural introduction of seminal fluid into the vulvovaginal environment, a pulse of artificial seminal fluid (ASF) was found to exert an enduring potential to overcome the inhibitory efficacy of artificial vaginal fluid (AVF) on filamentation. These findings suggest that a transient but substantial change in the nutrient levels within the vulvovaginal environment during unprotected coitus can induce resident C. albicans cells to engage developmental programs associated with virulent growth.


Hsa_circ_0003998 promotes cell proliferation and invasion by targeting miR-326 in non-small cell lung cancer.
Circular RNAs represent a new class of noncoding RNAs involved in the development of cancer. However, little is known about their role in non-small cell lung cancer (NSCLC). We examined hsa_circ_0003998 levels in 60 NSCLC tissues by quantitative real-time polymerase chain reaction and analyzed the clinicopathologic significance of hsa_circ_0003998 expression. The effect of small interfering RNA-mediated hsa_circ_0003998 knockdown on proliferation and invasion was analyzed in A549 and H1299 cells in vitro. Moreover, the target genes of hsa_circ_0003998 were further explored by bioinformatic analysis, dual luciferase reporter assays, and rescue experiments. Hsa_circ_0003998 upregulation was associated with larger tumor size and lymph node metastasis and also correlated with shorter overall survival of NSCLC patients. Functional experiments showed knockdown of hsa_circ_0003998 restrained cell proliferation and invasion in NSCLC cells. In particular, hsa_circ_0003998 upregulated the expression of miR-326 target gene Notch1 through sponging miR-326. Furthermore, the tumor-inhibiting effect of hsa_circ_0003998 silencing was blocked by miR-326 inhibitor. hsa_circ_0003998/miR-326/Notch1 pathway regulates the progression of NSCLC.


Long-term therapy of essential tremor with propranolol.
In a double-blind crossover study 12 patients with essential tremor were treated with propranolol and a placebo; 8 improved with propranolol and 3 with the placebo; the degree of improvement with propranolol was greater. In a similar study with diazepam 5 of 12 improved with diazepam and 4 of 12 with the placebo; the degree of improvement was less than that achieved with propranolol. Response in 21 patients to treatment with propranolol for 2 to 4 years was excellent in 4, good in 4 and fair in 10; the condition of 1 was unchanged and that of 2, worse. Excellent response was maintained for as long as 4 years, but response tended to deteriorate with time if initially it was less than excellent. Response decreased with increasing age. No patient 60 years of age or older had an excellent response, and the four with an excellent response were under age 55, three being under age 35; all four had had their tremor less than 12 years. Patients with essential tremor should be given a 3-month trial of propranolol at 120 mg/d; if no significant response is seen the dose should be decreased, then the drug discontinued.


Antibody epitope profiling of the KSHV LANA protein using VirScan.
The humoral antibody response against Kaposi sarcoma-associated herpesvirus (KSHV) in infected individuals has been characterized demonstrating the latency-associated nuclear antigen (LANA) as the most antigenic KSHV protein. Despite the antigenicity of the protein, specific LANA epitopes have not been systematically characterized. Here, we utilized a bacteriophage T7 library, which displays 56-amino acid KSHV LANA peptides with 28-amino acid overlap (VirScan), to define those epitopes in LANA targeted by antibodies from a cohort of 62 sub-Saharan African Kaposi sarcoma (KS) patients and 22 KSHV-infected asymptomatic controls. Intra- and inter-patient breadth and magnitude of the anti-LANA responses were quantified at the peptide and amino acid levels. From these data, we derived a detailed epitope annotation of the entire LANA protein, with a high-resolution focus on the N- and C-termini. Overall, the central repeat region was highly antigenic, but the responses to this region could not be confidently mapped due to its high variability. The highly conserved N-terminus was targeted with low breadth and magnitude. In a minority of individuals, antibodies specific to the nuclear localization sequence and a portion of the proline-rich regions of the N-terminus were evident. In contrast, the first half of the conserved C-terminal domain was consistently targeted with high magnitude. Unfortunately, this region was not included in LANA partial C-terminal crystal structures, however, it was predicted to adopt predominantly random-coil structure. Coupled with functional and secondary structure domain predictions, VirScan revealed fine resolution epitope mapping of the N- and C-terminal domains of LANA that is consistent with previous antigenicity studies and may prove useful to correlate KSHV humoral immunity with pathogenesis.


Re-expansion, re-oxygenation, and rethinking.
In a 1902 American Journal of the Medical Sciences case report, Riesman described "albuminous expectoration" following thoracentesis, a phenomenon that is now recognized as re-expansion pulmonary edema (RPE). Both cellular and biochemical mechanisms that produce lung injury in RPE have been described recently. Pathophysiologically, this unilateral edematous lung injury resembles the adult respiratory distress syndrome (ARDS) because both are characterized by intra-alveolar-activated neutrophils and markedly increased lung capillary permeability. Biochemical mechanisms that operate in RPE are analogous to those in diverse re-oxygenation (reperfusion) injuries that have been described recently in the heart, kidney, brain, and intestine. Re-oxygenated lung tissue appears to produce excess superoxide and other cytotoxic oxygen metabolites, although lung xanthine oxidase, the commonly recognized source of these oxidants, is exceedingly low. Riesman's critical analyses of the re-expansion edema fluid in his case provided an impetus for others to hypothesize that increased permeability pulmonary edema in RPE represented re-oxygenation injury of the lung microvasculature.


A comparison of fospropofol to midazolam for moderate sedation during outpatient dental procedures.
Moderate intravenous (IV) sedation combined with local anesthesia is common for outpatient oral surgery procedures. An ideal sedative agent must be safe and well tolerated by patients and practitioners. This study evaluated fospropofol, a relatively new sedative/hypnotic, in comparison to midazolam, a commonly used benzodiazepine, for IV moderate sedation during oral and maxillofacial surgery. Sixty patients were randomly assigned to either the fospropofol or the midazolam group. Each participant received 1 μg/kg of fentanyl prior to administration of the selected sedative. Those in the fospropofol group received an initial dose of 6.5 mg/kg, with 1.6 mg/kg supplemental doses as needed. Those in the midazolam group received initial doses of 0.05 mg/kg, followed by 0.02 mg/kg supplemental doses. The quality of sedation in each patient was evaluated with regard to (a) onset of sedation, maintenance, and recovery profile; (b) patient and surgeon satisfaction; and (c) hemodynamic stability and adverse effects. The fospropofol group demonstrated shorter physical recovery times than midazolam patients, taking a mean of 11.6 minutes versus 18.4 minutes for physical recovery (P = .007). Cognitive recovery comparison did not find any difference with a mean of 7.5 minutes versus 8.8 minutes between the 2 drug groups (P = .123). The fospropofol group had a higher rate of local anesthetic injection recall (90.5 vs 44.4%, P = .004). Other parameters of recall were comparable. Two adverse effects demonstrated significance, with more patients in the midazolam group experiencing tachycardia (48.2 vs 9.4%, P = .001), and more patients in the fospropofol group experiencing perineal discomfort (40.6 vs 0, P < .001). No significant difference was found in any other measures of sedation safety, maintenance, or satisfaction. Fospropofol, when administered intravenously by a dentist anesthesiologist at the indicated dose in this study, appears to be a safe, well-tolerated alternative to midazolam for intravenous moderate sedation during minor oral surgery procedures.


Oxygen uptake, heartrate and blood lactate responses to the Chito-Ryu Seisan kata in skilled karate practitioners.
To evaluate the efficacy of Seisan kata as an aerobic power training mode, four male (28.5 +/- 4.2 y) Chito-Ryu karate black belt practitioners did kata continuously for 10 min. In separate sessions the kata (formal, organized movement sequences) were done at rates of 1 (PACE) and 2 (FAST) kata "cycles" per minute. Heartrate (HR) and VO2 were monitored continuously during the sessions. VO2 during the PACE and FAST sessions averaged 73 +/- 3 and 94 +/- 2% of leg cycling VO2peak, respectively. The corresponding HRs were 93 +/- 6 and 101 +/- 3% of HRmax (leg cycle test). PACE and FAST post-exercise blood lactates were 12 +/- 4 and 22 +/- 6%, respectively, of the maximal leg cycle test values. These data indicate that karate kata can be used as an effective and specific means for training aerobic power in karate practitioners.


Functional Dynamics of Dorsal and Ventral Reading Networks in Bilinguals.
In today's world, bilingualism is increasingly common. However, it is still unclear how left-lateralized dorsal and ventral reading networks are tuned to reading in proficient second-language learners. Here, we investigated differences in functional regional activation and connectivity as a function of L1 and L2 reading, L2 orthographic depth, and task demands. Thirty-seven late bilinguals with the same L1 and either an opaque or transparent L2 performed perceptual and semantic reading tasks in L1 and L2 during functional magnetic resonance imaging (fMRI) scanning. Results revealed stronger regional recruitment for L2 versus L1 reading and stronger connectivity within the dorsal stream during L1 versus L2 reading. Differences in orthographic depth were associated with a segregated profile of left ventral occipitotemporal (vOT) coactivation with dorsal regions for the transparent L2 group and with ventral regions for the opaque L2 group. Finally, semantic versus perceptual demands modulated left vOT engagement, supporting the interactive account of the contribution of vOT to reading, and were associated with stronger coactivation within the ventral network. Our findings support a division of labor between ventral and dorsal reading networks, elucidating the critical role of the language used to read, L2 orthographic depth, and task demands on the functional dynamics of bilingual reading.


Monitoring hospital wastewaters for their probable genotoxicity.
Hospitals' effluents contain a considerable amount of chemicals. Considering the significant volume of wastewater discharged by hospitals, the presence of these chemicals represents a real threat to the environment and human health. Thus, the aim of this study was to evaluate the in vivo and in vitro genotoxicities of three wastewater effluents collected from Tunisian hospitals. The liver of Swiss albino male mice, previously treated with different doses of the hospital wastewaters, was used as a model to detect DNA fragmentation. Our results showed all the hospital effluents caused significant qualitative and quantitative hazards in hepatic DNA. The wastewater collected from Sfax hospital exhibited the highest genotoxic effect, which may be explained by the presence in this effluent of some toxic micropolluants. There was a significant increase in genotoxicity, proportionally to the concentration of effluent. However, the vitotox assay did not show any significant genotoxicity on Salmonella typhimurium TA104 in the presence or absence of microsomal fraction S9. The ratio gentox/cytox was lower than the threshold 1.5. This study assessed the toxicological risk issued from Tunisian hospital wastewaters, which is potentially very harmful, and it has been pointed out that wastewater treatment requires special attention.


Deregulation of the Genes that Are Involved in Drug Absorption, Distribution, Metabolism, and Excretion in Hepatocellular Carcinoma.
Genes involved in drug absorption, distribution, metabolism, and excretion (ADME) are called ADME genes. Currently, 298 genes that encode phase I and II drug metabolizing enzymes, transporters, and modifiers are designated as ADME genes by the PharmaADME Consortium. ADME genes are highly expressed in the liver and their levels can be influenced by liver diseases such as hepatocellular carcinoma (HCC). In this study, we obtained RNA-sequencing and microRNA (miRNA)-sequencing data from 371 HCC patients via The Cancer Genome Atlas liver hepatocellular carcinoma project and performed ADME gene-targeted differential gene expression analysis and expression correlation analysis. Two hundred thirty-three of the 298 ADME genes (78%) were expressed in HCC. Of these genes, almost one-quarter (58 genes) were significantly downregulated, while only 6% (15) were upregulated in HCC relative to healthy liver. Moreover, one-half (14/28) of the core ADME genes (CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2E1, CYP3A4, NAT1, NAT2, UGT2B7, SLC22A1, SLCO1B1, and SLCO1B3) were downregulated. In addition, about one-half of the core ADME genes were positively correlated with each other and were also positively (AHR, ARNT, HNF4A, PXR, CAR, PPARA, and RXRA) or negatively (PPARD and PPARG) correlated with transcription factors known as ADME modifiers. Finally, we show that most miRNAs known to regulate core ADME genes are upregulated in HCC. Collectively, these data reveal 1) an extensive transcription factor-mediated ADME coexpression network in the liver that efficiently coordinates the metabolism and elimination of endogenous and exogenous compounds; and 2) a widespread deregulation of this network in HCC, most likely due to deregulation of both transcriptional and post-transcriptional (miRNA) pathways.


PGT-HLA programmes for the cure of a sick sibling: clinical strategies for this challenging search.
The ultimate goal of a preimplantation genetic testing and human leukocyte antigen (PGT-HLA) matching programme is the birth of a healthy, HLA-compatible child for the treatment or cure of a sick sibling. Several authors have published successful cases of the births of children HLA-matched to siblings affected by different conditions and diseases. However, there are many reports of failed attempts. Couples seeking an HLA-matched sibling for their affected child look for positive outcomes in the shortest possible time. Nevertheless, there is no published consensus or guidelines with recommendations for these cases. Here, the authors aimed to analyse different approaches for these programmes, highlighting the most promising strategies for the families and fertility units. Furthermore, the authors mention a successful case of a PGT-HLA matching programme after a previous failed attempt following the strategies proposed. Which is the most cost-effective and time-efficient approach in a PGT-HLA matching programme?


Medical comorbidities in patients with serious mental illness: a retrospective study of mental health patients attending an outpatient clinic in Qatar.
The life span of individuals with serious mental illness (SMI) is shorter compared to the general population. This excess mortality is mainly due to physical illness. The aim of the study was to investigate the prevalence rates of different physical illnesses in individuals with SMI and to examine how these are being managed. The study was a cross-sectional retrospective chart review of a cohort of patients with SMI. A comprehensive electronic data extraction tool using SurveyMonkey® was used to collect patient demographics, psychiatric and medical comorbidities, medications and all relevant physical assessments. Data were then first extrapolated into an Excel® spreadsheet and later to SPSS® for data analysis. A descriptive statistical approach was used to analyze the demographic and clinical data. Chi-square test for categorical variables and t-test for continuous variables were used to compare the demographic and clinical characteristics of the cohort. A total of 336 patients with SMI were included for the retrospective chart review. The majority of these patients had a diagnosis of depression (50.3%), followed by schizophrenia (33.0%) and bipolar disorder (19.6%). Diabetes was the most frequent medical comorbidity, diagnosed in 16.1% of SMI patients, followed by hypertension (9.2%) and dyslipidemia (9.8%). Monitoring of comorbidity-associated risk factors and other relevant physical assessment parameters (such as blood pressure, weight, hemoglobin A1c [HbA1c], blood glucose and lipids) were documented in less than 50% of patients, and some parameters, such as smoking status, were not documented at all. Both, the literature and our cohort provide evidence that individuals with SMI are less likely to receive standard levels of care for their medical comorbidities.


Real-world evaluation of compliance and preference in Alzheimer's disease treatment: an observational study in Taiwan.
Among the medications approved for Alzheimer's disease (AD), rivastigmine is the only one available as transdermal patch. The aim of this study was to evaluate compliance and caregivers' preference with oral and transdermal (rivastigmine) monotherapy in patients with mild-to-moderate AD from Taiwan. Real-world Evaluation of Compliance And Preference in Alzheimer's disease treatment (RECAP) in Taiwan was a prospective, noninterventional, observational study with a 24-week (±8 weeks) observational period for each participant. Eligible patients were grouped into one of the two treatment cohorts based on the baseline AD therapy: oral (donepezil, galantamine, rivastigmine, or memantine) or transdermal (rivastigmine patch). The primary end points were caregiver preference and caregiver assessment of patients' compliance to the current medication (oral or transdermal medication) at Week 24 (end of the study). Safety was assessed by recording any adverse events. A total of 301 patients (age: 77.6±7.19 years) were enrolled from nine centers in Taiwan, of whom 138 (45.8%) patients were in the transdermal monotherapy cohort. Caregivers of patients who were exposed to both forms of therapies demonstrated a higher preference for transdermal rivastigmine monotherapy than the oral monotherapy (82.4% [n=61] versus 17.6% [n=13], P<0.0001); for patients treated with only one therapy, the caregivers' preference was significantly in favor of the treatment to which the patient was exposed (both P<0.0001). In both cohorts, patients showed good compliance, with an overall score of 8.65±1.38 on an 11-point scale. Of 301 enrolled patients, 102 (33.9%) reported at least one adverse event during the study (51 patients each in the two cohorts). With the higher caregiver preference and a good patient compliance, the trans-dermal rivastigmine patch is a suitable treatment choice for patients with mild-to-moderate AD, especially for patients intolerant to oral therapies.


Brain Transcriptomics of Wild and Domestic Rabbits Suggests That Changes in Dopamine Signaling and Ciliary Function Contributed to Evolution of Tameness.
Domestication has resulted in immense phenotypic changes in animals despite their relatively short evolutionary history. The European rabbit is one of the most recently domesticated animals, but exhibits distinct morphological, physiological, and behavioral differences from their wild conspecifics. A previous study revealed that sequence variants with striking allele frequency differences between wild and domestic rabbits were enriched in conserved noncoding regions, in the vicinity of genes involved in nervous system development. This suggests that a large proportion of the genetic changes targeted by selection during domestication might affect gene regulation. Here, we generated RNA-sequencing data for four brain regions (amygdala, hypothalamus, hippocampus, and parietal/temporal cortex) sampled at birth and revealed hundreds of differentially expressed genes (DEGs) between wild and domestic rabbits. DEGs in amygdala were significantly enriched for genes associated with dopaminergic function and all 12 DEGs in this category showed higher expression in domestic rabbits. DEGs in hippocampus were enriched for genes associated with ciliary function, all 21 genes in this category showed lower expression in domestic rabbits. These results indicate an important role of dopamine signaling and ciliary function in the evolution of tameness during rabbit domestication. Our study shows that gene expression in specific pathways has been profoundly altered during domestication, but that the majority of genes showing differential expression in this study have not been the direct targets of selection.


A Japanese Stretching Intervention Can Modify Lumbar Lordosis Curvature.
Eighteen healthy male adults were assigned to either an intervention or control group. Isogai dynamic therapy (IDT) is one of Japanese stretching interventions and has been practiced for over 70 years. However, its scientific quantitative evidence remains unestablished. The objective of this study was to determine whether IDT could modify lumbar curvature in healthy young adults compared with stretching exercises used currently in clinical practice. None of previous studies have provided data that conventional stretching interventions could modify spinal curvatures. However, this study provides the first evidence that a specific form of a Japanese stretching intervention can acutely modify the spinal curvatures. We compared the effects of IDT, a Japanese stretching intervention (n=9 males), with a conventional stretching routine (n=9 males) used widely in clinics to modify pelvic tilt and lumbar lordosis (LL) angle. We measured thoracic kyphosis (TK) and LL angles 3 times during erect standing using the Spinal Mouse before and after each intervention. IDT consisted of: (1) hip joint correction, (2) pelvic tilt correction, (3) lumbar alignment correction, and (4) squat exercise stretch. The control group performed hamstring stretches while (1) standing and (2) sitting. IDT increased LL angle to 25.1 degrees (±5.9) from 21.2 degrees (±6.9) (P=0.047) without changing TK angle (pretest: 36.8 degrees [±6.9]; posttest: 36.1 degrees [±6.5]) (P=0.572). The control group showed no changes in TK (P=0.819) and LL angles (P=0.744). IDT can thus be effective for increasing LL angle, hence anterior pelvic tilt. Such modifications could ameliorate low back pain and improve mobility in old adults with an unfavorable pelvic position.


[Results of treatment of retinal angiomatous proliferation with photodynamic therapy].
To determine the effects of phothodynamic therapy using verteporfin in the treatment of patients with retinal angiomatous proliferation (RAP) and the incidence of this retinal disease in our area. We performed a retrospective study of 11 cases of RAP who were treated with photodynamic therapy using verteporfin (PDT). The incidence of RAP in the group of eyes with minimally classic or occult subfoveal choroidal neovascularization was 8%. The mean follow-up time after treatment was 15 months. The visual acuity improved in 3, remained the same in 4, and decreased in 4. In 4 patients, angiomatous lesions were observed in both eyes. There is no method proven to be effective for the treatment of retinal angiomatous proliferation. Our results suggest that PDT may be useful therapy in patients with RAP as it appeared to reduce the risk of visual loss.


Parenting stress among child welfare involved families: Differences by child placement.
The intersection of parenting stress and maltreatment underscores the importance of understanding the factors associated with parenting stress among child welfare involved families. This study takes advantage of a statewide survey of child welfare involved families to examine parent and child characteristics and concrete resources, in relation to parenting stress. Separate multivariate analyses were conducted by placement status given the difference in day-to-day parenting responsibilities for families receiving in-home supervision compared to those whose children are in out-of-home care. Across both groups, parenting stress was predicted by child mental health, a finding with critical implications for intervention to this vulnerable group of families. Parent mental health also predicted parenting stress for the in-home group and food insecurity predicted parenting stress in the out-of-home group. Findings confirm that stress varies by context and that a multi-dimensional framework, considering both psychosocial and concrete resources, is required to capture contributors to parenting stress.


Natural compounds as next-generation herbicides.
Herbicides with new modes of action (MOAs) are badly needed due to the rapidly evolving resistance to commercial herbicides, but a new MOA has not been introduced in over 20 years. The greatest pest management challenge for organic agriculture is the lack of effective natural product herbicides. The structural diversity and evolved biological activity of natural phytotoxins offer opportunities for the development of both directly used natural compounds and synthetic herbicides with new target sites based on the structures of natural phytotoxins. Natural phytotoxins are also a source for the discovery of new herbicide target sites that can serve as the focus of traditional herbicide discovery efforts. There are many examples of strong natural phytotoxins with MOAs other than those used by commercial herbicides, which indicates that there are molecular targets of herbicides that can be added to the current repertoire of commercial herbicide MOAs.


Negative genetic correlation between traits of the Drosophila head, and interspecific divergence in head shape.
For morphological traits that are negatively correlated, the genetic correlation (rg) between them might strongly influence patterns of morphological divergence and shape. Here, the pattern of divergence between two sibling species of cactophilic Drosophila, D. buzzatii and D. koepferae, is examined for two traits that are known to be negatively correlated in other Drosophila species: face width (FW) and width of both eyes (EW). Head width (HW, the sum of FW and EW, i. e. the total width of the head capsule) was also examined. Genetic and phenotypic correlations were estimated in the laboratory G2 generation of a sample of wild D. buzzatii derived from a population where D. koepferae is not present. Phenotypic correlations were also estimated in D. buzzatii and D. koepferae from another, very different, population where the species are sympatric. Consistent with studies in other Drosophila species, rg was negative and significant for the correlation between FW and EW, and positive (but nonsignificant at a matrix-wide P-value of 0.05) for the correlations of HW with both FW and EW. This well-defined correlation pattern was also consistent with the phenotypic correlations in both D. buzzatii and D. koepferae. No significant difference in these traits was detected between D. buzzatii populations, but head shape has diverged between D. buzzatii and D. koepferae. Specifically, the two negatively correlated traits, FW and EW, have evolved in opposite directions in these two species, with HW showing no significant interspecific difference. The overall picture of this divergence pattern shows a striking concordance with the present evidence of negative correlations between FW and EW, and is consistent with the notion of rg-related constraints on the pattern of interspecific differentiation.


Exploring the reasons for orthopedic implant failure in traumatic fractures of the lower limb.
A damaged orthopedic implant in the body can cause problems for the patient and disrupt the therapeutic process. This study investigates various factors for orthopedic implant failures in patients with traumatic lower limb fractures who referred to a trauma center in Mashhad, Iran. This was a prospective study of 23 patients referred to Kamyab University Hospital with failed orthopedic implants in 2009. We included any patient with an orthopedic device previously implanted secondary to a traumatic lower limb fracture who later presented to Kamyab University Hospital because of a failed implant. For all patients, a thorough history was taken and the necessary investigations that included radiographic studies were performed. We investigated the quality of the failed devices by chemical analysis, metal hardness testing and metallography. The results were statistically analyzed. The mean age of the patients was 33 ± 19 years. There were 19 (82.5%) male and 4 (17.4%) female patients. In 14 (60.9%) cases, there were failed femoral implants and 9 (39.1%) cases had failed implants for tibial fractures. We compared the implants against ASTM standards. According to chemical analysis, all internally produced devices and one of the leading international brands were within the expected standard. However, in 3 cases chemical analysis showed a deviation from the standards. These were manufactured by "miscellaneous" companies. In one (4.3%) case the device failure was iatrogenic, in 14 (60.9%) it was due to faulty implants and in 8 (34.8%) cases, the patients were non-compliant with instructions. Hardness testing was satisfactory in all cases and metallographic studies showed good quality for the leading international brands, satisfactory quality for the internally produced devices and poor quality for the miscellaneous devices. The implants classified as miscellaneous were of poor quality. Thus, we cannot recommend their use in orthopedic surgeries. We recommend using credible, known brands.


Anthropometric, parental, and psychosocial correlates of dietary intake of African-American girls.
This paper identifies the anthropometric, parental, and psychosocial characteristics and meal practices (e.g., breakfast skipping and number of meals and snacks consumed) associated with consumption of total energy, percent energy from fat, fruit, 100% fruit juice, vegetables, sweetened beverages, and water among 8- to 10-year-old African-American girls. This study included 114 8- to 10-year-old African-American girls and a parent or primary caregiver. Girls and a parent or primary caregiver completed several dietary questionnaires. Two 24-hour dietary recalls were conducted with each girl. Height and weight were measured. Separate hierarchical regression analyses were conducted for each dependent dietary variable; potential field center differences were examined. The number of meals and snacks consumed was correlated with energy intake. Lower BMI was related to higher vegetable consumption, and the number of snacks consumed was positively related to sweetened beverage consumption. Greater low-fat food preparation practices reported by parents were related to lower consumption of fat as a percentage of total energy. Dietary behavior differed across geographic areas. Low-fat food preparation practices in the home seemed to be an important influence on the percentage of energy consumed from fat. Greater vegetable consumption was associated with lower BMI. Interventions to prevent excessive weight gain in African-American girls should encourage low-fat food preparation in the home and greater consumption of vegetables.


Delays, interruptions, and losses from prevention of mother-to-child transmission of HIV services during antenatal care in Johannesburg, South Africa: a cohort analysis.
Between 2010-2013, South Africa implemented WHO 'Option A' for prevention of mother to child transmission (PMTCT), where all HIV-infected pregnant women (from 14 weeks gestation) received zidovudine (AZT) as ARV prophylaxis and initiated CD4 testing at their first antenatal care (ANC) visit. After returning for a second visit to collect CD4 results, women with CD4 counts ≤ 350 were referred to the ART clinic and fast-tracked for initiation on lifelong ART while continuing to visit the ANC clinic every four weeks. Women with CD4 counts >350 were dispensed daily AZT prophylaxis at monthly follow up visits (every 4 weeks). The primary objective of this study was to evaluate adherence of HIV-infected pregnant women to recommended PMTCT services at and after their first antenatal care (ANC) visit. We conducted an observational cohort study from August 2012 to February 2013 at two primary health care clinics in Johannesburg, South Africa using routinely collected clinic data from first ANC visit for up to 60 days. Of the 158 patients newly diagnosed with HIV at their first ANC visit, records indicated that 139 women initiated CD4 testing during their first ANC visit. 52 patients (33% of 158) did not return again to the clinic within 60 days. Of the 118 (84% of 139) women with known gestational age > 13 weeks and known Hb ≥ 8 g/dl who should have received a 4-week supply of daily AZT at first ANC visit, 81 women (69% of 118) had a record of AZT being dispensed. Among the 139 women with CD4 results, 72 (52%) were eligible for lifelong ART (CD4 count ≤350); however, only 2 initiated ART within 30 days. Loss to initiation of both single and triple ARV therapy, loss to follow-up, and treatment interruptions were common during ANC care for pregnant women with HIV after their first ANC visit.


Predictors of Engagement, Response to Follow Up, and Extent of Alcohol Reduction in Users of a Smartphone App (Drink Less): Secondary Analysis of a Factorial Randomized Controlled Trial.
Digital interventions for alcohol can help achieve reductions in hazardous and harmful alcohol consumption. The Drink Less app was developed using evidence and theory, and a factorial randomized controlled trial (RCT) suggested that 4 of its intervention modules may assist with drinking reduction. However, low engagement is an important barrier to effectiveness, and low response to follow up is a challenge for intervention evaluation. Research is needed to understand what factors influence users' level of engagement, response to follow up, and extent of alcohol reduction. This study aimed to investigate associations between user characteristics, engagement, response to follow up, and extent of alcohol reduction in an app-based intervention, Drink Less. This study involved a secondary data analysis of a factorial RCT of the Drink Less app. Participants (N=672) were aged 18 years or older, lived in the United Kingdom, and had an Alcohol Use Disorders Identification Test score >7 (indicative of excessive drinking). Sociodemographic and drinking characteristics were assessed at baseline. Engagement was assessed in the first month of use (number of sessions, time on app, number of days used, and percentage of available screens viewed). Response to follow up and extent of alcohol reduction (change in past week consumption) were measured after 1 month. Associations were assessed using unadjusted and adjusted linear or logistic regression models. Age (all unstandardized regression coefficients [B] >.02, all P<.001) and post-16 educational qualifications (all B>.18, all P<.03) were positively associated with all engagement outcomes. Age (odds ratio [OR] 1.04, P<.001), educational qualifications (OR 2.11, P<.001), and female gender (OR 1.58, P=.02) were positively associated with response to follow up. Engagement outcomes predicted response to follow up (all OR>1.02, all P<.001) but not the extent of alcohol reduction (all -.14<B<-.06, all P>.07). Baseline drinking characteristics were the only variables associated with the extent of alcohol reduction among those followed up (all B>.49, all P<.001). Users of the alcohol reduction app, Drink Less, who were older and had post-16 educational qualifications engaged more and were more likely to respond at 1-month follow up. Higher baseline alcohol consumption predicted a greater extent of alcohol reduction among those followed up but did not predict engagement or response to follow up. Engagement was associated with response to follow up but was not associated with the extent of alcohol reduction, which suggests that the Drink Less app does not have a dose-response effect. International Standard Randomised Controlled Trial Number ISRCTN40104069; http://www.isrctn.com/ISRCTN40104069 (Archived by WebCite at http://www.webcitation.org/746HqygIV).


A rare case of cerebellar degeneration due to primary Sjogren's syndrome.
Sjogren's syndrome (SS) is an autoimmune pathology which mainly affects salivary and lacrimal glands. Cerebellar degeneration association with SS is very rare, with only a few cases described. The treatment of SS with central nervous system involvement is not consensual. We present a 48-year-old woman with dysarthria, diplopia and ataxia associated with xerostomia. The brain magnetic imaging revealed bilateral cerebellar atrophy. She had antibody positivity for anti-SSA and anti-SSB and minor salivary glands biopsy revealed lymphocytic infiltration. Methylprednisolone, cyclophosphamide and intravenous immunoglobulin yielded no results. Rituximab was initiated with improvement in dysarthria and coordination.


Presence and localization of two vitamin D-dependent calcium binding proteins in kidneys of higher vertebrates.
We looked for the presence of vitamin D-dependent intestinal calcium binding protein (CaBP), relative molecular mass (mol wt) about 10,000, in kidneys of mammals, birds, and reptiles, and compared the localization of this 10,000 mol wt CaBP with the localization of the 28,000 mol wt vitamin D-dependent CaBP. Antiserum directed against rat intestinal CaBP (RICaBP) was used with the unlabeled antibody peroxidase-antiperoxidase technique to localize the 10,000 mol wt CaBP. Kidneys of adult Swiss and DBA/2J mice were positive for the 10,000 mol wt CaBP, whereas kidneys of adult rat, chicken, and two species of lizard were negative. Extracts of adult rat kidney showed no detectable immunoreactivity with the antiserum to RICaBP by radial immunodiffusion assay. However, kidneys of postnatal rats (12 days old) exhibited limited immunocytochemical reactivity and relatively low immunoreactivity (about 1 micrograms/mg protein) for 10,000 mol wt CaBP. In both strains of mice, 10,000 mol wt CaBP was localized predominantly to the distal convoluted tubules, connecting tubules, and collecting tubules of the cortical labyrinth. In 12-day-old rats 10,000 mol wt CaBP was localized to the distal convoluted tubules and cortical ascending thick limbs. Absorption of the antiserum with electrophoretically pure RICaBP eliminated specific reactivity. By dual color staining of mouse kidneys, the population of cells reactive for the 10,000 mol wt CaBP was found to be similar, but not identical, to that population of cells reactive for the 28,000 mol wt CaBP. Absorption of the antiserum to RICaBP with either rat renal CaBP or chicken intestinal CaBP (both 28,000 mol wt CaBPs) had no affect on the positive reactivity of the distal mouse nephron for RICaBP. Conversely, absorption of the antiserum to rat renal CaBP with the 10,000 mol wt RICaBP had no affect on reactivity for the 28,000 mol wt CaBP. Thus, two immunologically distinct CaBPs, mol wt 10,000 and 28,000, are present in the adult mouse nephron; whereas, kidneys of adult rats, chickens, and saurian reptiles apparently contain significant levels of only 28,000 mol wt CaBP. Since the 12-day-old rat nephron contains both the 28,000 mol wt and the 10,000 mol wt CaBP it appears that in the kidney, ontogenetically, and perhaps evolutionarily as well, the 28,000 mol wt CaBP is more highly conserved.


Staff recognition and its importance for surgical service delivery: a qualitative study in Freetown, Sierra Leone.
We examined the views of providers and users of the surgical system in Freetown, Sierra Leone on processes of care, job and service satisfaction and barriers to achieving quality and accessible care, focusing particularly on the main public tertiary hospital in Freetown and two secondary and six primary sites from which patients are referred to it. We conducted interviews with health care providers (N = 66), service users (n = 24) and people with a surgical condition who had chosen not to use the public surgical system (N = 13), plus two focus groups with health providers in primary care (N = 10 and N = 10). The overall purpose of the study was to understand perceptions on processes of and barriers to care from a variety of perspectives, to recommend interventions to improve access and quality of care as part of a larger study. Our research suggests that providers perceive their relationships with patients to be positive, while the majority of patients see the opposite: that many health workers are unapproachable and uncaring, particularly towards poorer patients who are unable or unwilling to pay staff extra in the form of informal payments for their care. Many health care providers note the importance of lack of recognition shown to them by their superiors and the health system in general. We suggest that this lack of recognition underlies poor morale, leading to poor care. Any intervention to improve the system should therefore consider staff-patient relations as a key element in its design and implementation, and ideally be led and supported by frontline healthcare workers.


Housing conditions and respiratory morbidity in Indigenous children in remote communities in Northwestern Ontario, Canada.
Rates of lower respiratory tract infection (LRTI) among First Nations (FN) children living in Canada are elevated. We aimed to quantify indoor environmental quality (IEQ) in the homes of FN children in isolated communities and evaluate any associations with respiratory morbidity. We performed a cross-sectional evaluation of 98 FN children (81 with complete data) aged 3 years or younger, living in 4 FN communities in the Sioux Lookout region of Northern Ontario. We performed medical chart reviews and administered questionnaires. We performed a housing inspection, including quantifying the interior surface area of mould (SAM). We monitored air quality for 5 days in each home and quantified the contaminant loading of settled floor dust, including endotoxin. We analyzed associations between IEQ variables and respiratory conditions using univariable and multivariable analyses. Participants had a mean age of 1.6 years and 21% had been admitted to hospital for respiratory infections before age 2 years. Houses were generally crowded (mean occupancy 6.6 [standard deviation 2.6, range 3-17] people per house). Serious housing concerns were frequent, including a lack of functioning controlled ventilation. The mean SAM in the occupied space was 0.2 m2. In multivariable modelling, there was evidence of an association of LRTI with log endotoxin (p = 0.07) and age (p = 0.02), and for upper respiratory tract infections, with SAM (p = 0.07) and age (p = 0.03). Wheeze with colds was associated with log endotoxin (p = 0.03) and age (p = 0.04). We observed poor housing conditions and an association between endotoxin and wheezing in young FN children living in Northern Ontario.


Limited genomic consequences of hybridization between two African clawed frogs, Xenopus gilli and X. laevis (Anura: Pipidae).
The Cape platanna, Xenopus gilli, an endangered frog, hybridizes with the African clawed frog, X. laevis, in South Africa. Estimates of the extent of gene flow between these species range from pervasive to rare. Efforts have been made in the last 30 years to minimize hybridization between these two species in the west population of X. gilli, but not the east populations. To further explore the impact of hybridization and the efforts to minimize it, we examined molecular variation in one mitochondrial and 13 nuclear genes in genetic samples collected recently (2013) and also over two decades ago (1994). Despite the presence of F 1 hybrids, none of the genomic regions we surveyed had evidence of gene flow between these species, indicating a lack of extensive introgression. Additionally we found no significant effect of sampling time on genetic diversity of populations of each species. Thus, we speculate that F 1 hybrids have low fitness and are not backcrossing with the parental species to an appreciable degree. Within X. gilli, evidence for gene flow was recovered between eastern and western populations, a finding that has implications for conservation management of this species and its threatened habitat.


Adipocyte PU.1 knockout promotes insulin sensitivity in HFD-fed obese mice.
Insulin resistance is a key feature of obesity and type 2 diabetes. PU.1 is a master transcription factor predominantly expressed in macrophages but after HFD feeding PU.1 expression is also significantly increased in adipocytes. We generated adipocyte specific PU.1 knockout mice using adiponectin cre to investigate the role of PU.1 in adipocyte biology, insulin and glucose homeostasis. In HFD-fed obese mice systemic glucose tolerance and insulin sensitivity were improved in PU.1 AKO mice and clamp studies indicated improvements in both adipose and liver insulin sensitivity. At the level of adipose tissue, macrophage infiltration and inflammation was decreased and glucose uptake was increased in PU.1 AKO mice compared with controls. While PU.1 deletion in adipocytes did not affect the gene expression of PPARg itself, we observed increased expression of PPARg target genes in eWAT from HFD fed PU.1 AKO mice compared with controls. Furthermore, we observed decreased phosphorylation at serine 273 in PU.1 AKO mice compared with fl/fl controls, indicating that PPARg is more active when PU.1 expression is reduced in adipocytes. Therefore, in obesity the increased expression of PU.1 in adipocytes modifies the adipocyte PPARg cistrome resulting in impaired glucose tolerance and insulin sensitivity.


Heterozygous frameshift variants in HNRNPA2B1 cause early-onset oculopharyngeal muscular dystrophy.
Missense variants in RNA-binding proteins (RBPs) underlie a spectrum of disease phenotypes, including amyotrophic lateral sclerosis, frontotemporal dementia, and inclusion body myopathy. Here, we present ten independent families with a severe, progressive muscular dystrophy, reminiscent of oculopharyngeal muscular dystrophy (OPMD) but of much earlier onset, caused by heterozygous frameshift variants in the RBP hnRNPA2/B1. All disease-causing frameshift mutations abolish the native stop codon and extend the reading frame, creating novel transcripts that escape nonsense-mediated decay and are translated to produce hnRNPA2/B1 protein with the same neomorphic C-terminal sequence. In contrast to previously reported disease-causing missense variants in HNRNPA2B1, these frameshift variants do not increase the propensity of hnRNPA2 protein to fibrillize. Rather, the frameshift variants have reduced affinity for the nuclear import receptor karyopherin β2, resulting in cytoplasmic accumulation of hnRNPA2 protein in cells and in animal models that recapitulate the human pathology. Thus, we expand the phenotypes associated with HNRNPA2B1 to include an early-onset form of OPMD caused by frameshift variants that alter its nucleocytoplasmic transport dynamics.


Role of lipid peroxidation in post-traumatic spinal cord degeneration: a review.
A large amount of biochemical, physiological, and pharmacological data has been obtained which supports a mechanistic role of oxygen free radical-induced lipid peroxidation (LP) in post-traumatic spinal cord degeneration. Biochemical evidence of early and progressive lipid peroxidative reactions occurring in the injured spinal cord includes: an increase in polyunsaturated fatty acid peroxidation products (e.g., malonyldialdehyde), a decrease in cholesterol and the appearance of cholesterol oxidation products, an increase in cyclic GMP presumably due to free radical activation of guanylate cyclase, a decrease in tissue anti-oxidant levels (e.g., alpha tocopherol, reduced ascorbate), and inhibition of membrane-bound enzymes such as Na+ + K+-ATPase. In vitro CNS tissue studies have provided support for the possibility that LP may contribute to other early post-traumatic events including intracellular calcium accumulation and arachidonic acid release. Moreover, spinal tissue lactic acidosis, which occurs early after injury, can exacerbate LP reactions. The involvement of LP in the development of progressive post-traumatic spinal white matter ischemia has been strongly inferred from pharmacological studies in cats with known inhibitors of LP. For example, the dose-response curves for the ability of the glucocorticoid methylprednisolone (MP) to inhibit post-traumatic LP and to retard ischemia development are identical. This relationship between LP and post-traumatic ischemia is more directly implied from studies showing that pretreatment of cats with high doses of anti-oxidants (e.g., d-alpha tocopherol plus selenium p.o. or 1-ascorbic acid i.v.) can also significantly antagonize the progressive decrease in spinal cord blood flow that follows severe blunt injury. However, a similar efficacy of certain calcium and prostaglandin antagonists suggests an interrelationship between aberrant calcium fluxes, vasoconstrictor/platelet aggregating prostanoids, and LP in the post-traumatic ischemic phenomenon. In addition to a role of LP in ischemia development, the action of intensive d-alpha tocopherol and selenium pretreatment to retard anterograde cat motor nerve fiber degeneration after nerve section suggests that LP may also be a fundamental mechanism of "Wallerian" axonal degeneration after neural injury. Finally, a critical role of LP in the acute pathophysiology of CNS injury in general has been supported by the finding of an excellent correlation, in terms of efficacy and potency, between the action of glucocorticoid and nonglucocorticoid steroids to inhibit neural tissue LP in vitro and to promote early neurological recovery in severely head-injured mice.


Bacteria capture, concentration and detection by alternating current dielectrophoresis and self-assembly of dispersed single-wall carbon nanotubes.
The high polarizability and dielectrophoretic mobility of single-walled carbon nanotubes (SWNT) are utilized to capture and detect low numbers of bacteria and submicron particles in milliliter-sized samples. Concentrated SWNT solutions are mixed with the sample and a high-frequency (>100 kHz) alternating current (AC) field is applied by a microelectrode array to enhance bulk absorption of the particles (bacteria and nanoparticle substitutes) by the SWNTs via dipole-dipole interaction. The same AC field then drives the SWNT-bacteria aggregates to the microelectrode array by positive-AC dielectrophoresis (DEP), with enhanced and reversed bacteria DEP mobility due to the attached SWNTs. Since the field frequency exceeds the inverse RC time of the electrode double layer, the AC field penetrates deeply into the bulk and across the electrode gap. Consequently, the SWNTs and absorbed bacteria assemble rapidly (<5 min) into conducting linear aggregates between the electrodes. Measured AC impedance spectra by the same trapping electrodes and fields show a detection threshold of 10(4) bacteria/mL with this pathogen trapping and concentration technique.


Direct Growth of Carbon Nanotubes on New High-Density 3D Pyramid-Shaped Microelectrode Arrays for Brain-Machine Interfaces.
Silicon micromachined, high-density, pyramid-shaped neural microelectrode arrays (MEAs) have been designed and fabricated for intracortical 3D recording and stimulation. The novel architecture of this MEA has made it unique among the currently available micromachined electrode arrays, as it has provided higher density contacts between the electrodes and targeted neural tissue facilitating recording from different depths of the brain. Our novel masking technique enhances uniform tip-exposure for variable-height electrodes and improves process time and cost significantly. The tips of the electrodes have been coated with platinum (Pt). We have reported for the first time a selective direct growth of carbon nanotubes (CNTs) on the tips of 3D MEAs using the Pt coating as a catalyzer. The average impedance of the CNT-coated electrodes at 1 kHz is 14 kΩ. The CNT coating led to a 5-fold decrease of the impedance and a 600-fold increase in charge transfer compared with the Pt electrode.


Partitioning neuronal variability.
Responses of sensory neurons differ across repeated measurements. This variability is usually treated as stochasticity arising within neurons or neural circuits. However, some portion of the variability arises from fluctuations in excitability due to factors that are not purely sensory, such as arousal, attention and adaptation. To isolate these fluctuations, we developed a model in which spikes are generated by a Poisson process whose rate is the product of a drive that is sensory in origin and a gain summarizing stimulus-independent modulatory influences on excitability. This model provides an accurate account of response distributions of visual neurons in macaque lateral geniculate nucleus and cortical areas V1, V2 and MT, revealing that variability originates in large part from excitability fluctuations that are correlated over time and between neurons, and that increase in strength along the visual pathway. The model provides a parsimonious explanation for observed systematic dependencies of response variability and covariability on firing rate.


Pro108Ser mutation of SARS-CoV-2 3CLpro reduces the enzyme activity and ameliorates the clinical severity of COVID-19.
Recently, an international randomized controlled clinical trial showed that patients with SARS-CoV-2 infection treated orally with the 3-chymotrypsin-like protease (3CLpro) inhibitor PF-07321332 within three days of symptom onset showed an 89% lower risk of COVID-19-related hospital admission/ death from any cause as compared with the patients who received placebo. Lending support to this critically important result of the aforementioned trial, we demonstrated in our study that patients infected with a SARS-Cov-2 sub-lineage (B.1.1.284) carrying the Pro108Ser mutation in 3CLpro tended to have a comparatively milder clinical course (i.e., a smaller proportion of patients required oxygen supplementation during the clinical course) than patients infected with the same sub-lineage of virus not carrying the mutation. Characterization of the mutant 3CLpro revealed that the Kcat/Km of the 3CLpro enzyme containing Ser108 was 58% lower than that of Pro108 3CLpro. Hydrogen/deuterium-exchange mass spectrometry (HDX-MS) revealed that the reduced activity was associated with structural perturbation surrounding the substrate-binding region of the enzyme, which is positioned behind and distant from the 108th amino acid residue. Our findings of the attenuated clinical course of COVID-19 in patients infected with SARS-CoV-2 strains with reduced 3CLpro enzymatic activity greatly endorses the promising result of the aforementioned clinical trial of the 3CLpro inhibitor.


Liquid chromatographic determination of para-toluenesulfonamide in edible fillet tissues from three species of fish.
Chloramine-T (N-sodium-N-chloro-p-toluene-sulfonamide) is a candidate therapeutic drug for treating bacterial gill disease, a predominant disease of a variety of fish species. Research has been initiated to obtain the U.S. Food and Drug Administration's (FDA) approval for the use of chloramine-T on a variety of fish species. An attribute of a therapeutic aquaculture drug that must be characterized before the FDA approves its use is depletion of the drug's marker residue (the drug's parent compound or metabolite of highest concentration in an edible tissue). para-Toluenesulfonamide (p-TSA) is the primary degradation product and marker residue for chloramine-T in rainbow trout. To conduct residue depletion studies for chloramine-T in fish, a robust analytical method sensitive and specific for p-TSA residues in edible fillet tissue from a variety of fish was required. Homogenized fillet tissues from rainbow trout (Oncorhynchus mykiss), walleye (Stizostedion vitreum), and channel catfish (Ictalurus punctatus) were fortified at nominal p-TSA concentrations of 17, 67, 200, 333, and 1000 ng/g. Samples were analyzed by isocratic reversed-phase liquid chromatography (LC) with absorbance detection at 226 nm. Mean recoveries of p-TSA ranged from 77 to 93.17%; relative standard deviations ranged from 1.5 to 14%; method quantitation limits ranged from 13 to 18 ng/g; and method detection limits ranged from 3.8 to 5.2 ng/g. The LC parameters produced p-TSA peaks without coelution of endogenous compounds and excluded chromatographic interference from at least 20 chemicals and drugs of potential use in aquaculture.


Durable immunity to SARS-CoV-2 in both lower and upper airways achieved with a gorilla adenovirus (GRAd) S-2P vaccine in non-human primates.
SARS-CoV-2 continues to pose a global threat, and current vaccines, while effective against severe illness, fall short in preventing transmission. To address this challenge, there's a need for vaccines that induce mucosal immunity and can rapidly control the virus. In this study, we demonstrate that a single immunization with a novel gorilla adenovirus-based vaccine (GRAd) carrying the pre-fusion stabilized Spike protein (S-2P) in non-human primates provided protective immunity for over one year against the BA.5 variant of SARS-CoV-2. A prime-boost regimen using GRAd followed by adjuvanted S-2P (GRAd+S-2P) accelerated viral clearance in both the lower and upper airways. GRAd delivered via aerosol (GRAd(AE)+S-2P) modestly improved protection compared to its matched intramuscular regimen, but showed dramatically superior boosting by mRNA and, importantly, total virus clearance in the upper airway by day 4 post infection. GrAd vaccination regimens elicited robust and durable systemic and mucosal antibody responses to multiple SARS-CoV-2 variants, but only GRAd(AE)+S-2P generated long-lasting T cell responses in the lung. This research underscores the flexibility of the GRAd vaccine platform to provide durable immunity against SARS-CoV-2 in both the lower and upper airways.