Differential regulation of the Proteus mirabilis urease gene cluster by UreR and H-NS.
Proteus mirabilis, a cause of catheter-associated urinary tract infection, relies on several virulence factors to colonize the urinary tract. Among these, urease contributes to the development of urinary stones resulting from the increase in local pH due to urease-mediated hydrolysis of urea to NH(3) and CO(2). UreR, an AraC-like transcriptional activator, activates transcription of the genes encoding the urease subunits and accessory proteins (ureDABCEFG) in the presence of urea. UreR also initiates transcription of its own gene in a urea-inducible manner by binding to the intergenic region between ureR and ureD. The intergenic region contains poly(A) tracts that appear to be the target of H-NS. It has been shown that Escherichia coli and P. mirabilis H-NS acts to repress transcription of ureR in an E. coli model system. It was hypothesized that H-NS represses urease gene expression in the absence of UreR and urea by binding to the intergenic region. To demonstrate this the P. mirabilis hns gene was cloned and the 15.6 kDa H-NS was overexpressed and purified as a myc-His tail fusion. Using a gel shift assay, purified H-NS-myc-His bound preferentially to a 609 bp DNA fragment containing the entire ureR-ureD intergenic region. H-NS and UreR were able to displace each other from the ureR-ureD intergenic region. Circular permutation analysis revealed that the intergenic region is bent. Moreover, H-NS recognizes this curvature, binds the DNA fragment and induces further bending of the DNA as shown by a circular ligation assay. The effects of H-NS, urea and temperature (25 vs 37 degrees C) on urease expression were shown in E. coli containing an hns knockout and P. mirabilis where expression was increased at 37 degrees C. Increased transcription from p(ureR) was seen in the E. coli hns knockout when temperature was increased from 25 to 37 degrees C. These findings suggest H-NS and UreR differentially regulate urease in a negative and positive manner, respectively.


Testing a Positive Psychological Intervention for Osteoarthritis.
Osteoarthritis is a leading cause of disability for which there is no cure. Psychosocial-oriented treatments are underexplored. We developed and tested an intervention to build positive psychological skills (e.g., gratitude) to reduce osteoarthritis symptom severity, including pain and functioning, and to improve psychosocial well-being in patients with knee or hip osteoarthritis. Two-arm randomized design with six-month follow-up. An academic Veterans Affairs Medical Center. Patients aged 50 years or older with knee or hip osteoarthritis and pain ratings of 4 or higher. Patients (N = 42) were randomized to a six-week program containing positive skill-building activities or neutral control activities tailored to the patient population. Adherence was assessed by telephone each week. We assessed osteoarthritis symptom severity (WOMAC Osteoarthritis Index) and measures of well-being (positive affect, negative affect, and life satisfaction) at baseline and by telephone one, three, and six months after the program ended. We used linear mixed models to examine changes over time. The majority (64%) of patients completed more than 80% of their weekly activities. Patients in the positive (vs neutral) program reported significantly more improvement over time in osteoarthritis symptom severity (P = 0.02, Cohen's d = 0.86), negative affect (P = 0.03, Cohen's d = 0.50), and life satisfaction (P = 0.02, Cohen's d = 0.36). The study successfully engaged patients with knee or hip osteoarthritis in a six-week intervention to build positive psychological skills. Improving osteoarthritis symptom severity and measures of psychosocial well-being, the intervention shows promise as a tool for chronic pain management.


The expression of TTF1, CDX2 and ISL1 in 74 poorly differentiated neuroendocrine carcinomas.
The expression profile of immunohistochemical markers of origin in poorly differentiated neuroendocrine carcinoma (PDNEC) is not well studied. Seventy-four PDNECs from gastroenteropancreatic (GEP) organs and the lung, including 48 large cell NEC (LCNEC) and 26 small cell carcinomas (SmCC), were subject to immunohistochemical staining for CDX2, TTF1 and ISL1. The staining intensity (1 to 3) and percentage of positive tumor cells [0 (negative), 1 (<50%) and 2 (≥50%)] were assessed. The multiplicative index (maximum 6) was calculated and the average total score (aTS) was determined for each primary site and histologic subtype. In the 38 GEP and 36 lung PDNECs, CDX2, TTF1 and ISL1 staining was observed in 71% (aTS 2.8), 16% (aTS 0.4), 63% (aTS 1.9), and 22% (aTS 0.6), 72% (aTS 2.9) and 92% (aTS 3.8), respectively. GEP PDNECs showed a higher aTS for CDX2 and lower aTS for TTF1 and ISL1, compared to that of lung PDNECs (Student's t-test, p < 0.001). SmCC had a higher aTS for TTF1 and ISL1 (p < 0.001) and lower aTS for CDX2 (p < 0.002) than that of LCNEC. CDX2 and TTF1 demonstrate potential utility in suggesting the primary site of PDNEC. In addition, CDX2 may be useful in supporting the diagnosis of LCNEC in cases with overlapping or borderline morphology. Utility of ISL1 as an adjunctive diagnostic marker of SmCC remains to be studied.


The Investigation of Bio-impedance Analysis at a Wrist Phantom with Two Pulsatile Arteries.
Bio-impedance analysis (BIA) has been widely investigated for hemodynamic monitoring. However, previous works rarely modelled two synchronously pulsatile arteries (representing the radial and ulnar arteries) in the wrist/forearm model. This work aims to clarify and quantify the influences of two pulsatile arteries on BIA. First, two blood-filled arteries were structured in a 3D wrist segment using the finite element method (FEM). Afterwards, an easy-to-produce two-arteries artificial wrist was fabricated with two components: gelatine-based surrounding tissue phantom and saline blood phantom. A syringe driver was utilised to constrict the arteries, and the impedance signals were measured using a Multi-frequency Impedance Analyser (MFIA). Both simulation and experimental results demonstrated the non-negligible influences of the ulnar artery on the overall BIA, inducing unwanted resistance changes to the acquired signals from the radial artery. The phantom experiments revealed the summation of the individual resistance changes caused by a single pulsatile artery was approximately equal to the measured resistance change caused by two synchronously pulsatile arteries, confirming the measured impedance signal at the wrist contains the pulsatile information from both arteries. This work is the first simulation and phantom investigation into two synchronously pulsatile arteries under BIA in the distal forearm, providing a better insight and understanding in the morphology of measured impedance signals. Future research can accordingly select either a small spacing 4-spot electrode configuration for a single artery sensing or a band electrode configuration for overall pulsatile arteries sensing. A more accurate estimation of blood volume change and pulse wave analysis (PWA) could help to develop cuffless blood pressure measurement (BPM).


A PCR amplicon-based SARS-CoV-2 replicon for antiviral evaluation.
The development of specific antiviral compounds to SARS-CoV-2 is an urgent task. One of the obstacles for the antiviral development is the requirement of biocontainment because infectious SARS-CoV-2 must be handled in a biosafety level-3 laboratory. Replicon, a non-infectious self-replicative viral RNA, could be a safe and effective tool for antiviral evaluation. Herein, we generated a PCR-based SARS-CoV-2 replicon. Eight fragments covering the entire SARS-CoV-2 genome except S, E, and M genes were amplified with HiBiT-tag sequence by PCR. The amplicons were ligated and in vitro transcribed to RNA. The cells electroporated with the replicon RNA showed more than 3000 times higher luminescence than MOCK control cells at 24 h post-electroporation, indicating robust translation and RNA replication of the replicon. The replication was drastically inhibited by remdesivir, an RNA polymerase inhibitor for SARS-CoV-2. The IC50 of remdesivir in this study was 0.29 μM, generally consistent to the IC50 obtained using infectious SARS-CoV-2 in a previous study (0.77 μM). Taken together, this system could be applied to the safe and effective antiviral evaluation without using infectious SARS-CoV-2. Because this is a PCR-based and transient replicon system, further improvement including the establishment of stable cell line must be achieved.


Early glucose metabolism in children at risk for type 1 diabetes based on islet autoantibodies compared to low-risk control groups.
Anatomic variation or early differences in glucose metabolism have been linked to the development of type 1 diabetes. We aimed to describe early glucose metabolism based on HbA1c, oral glucose tolerance test (OGTT), and random plasma glucose years before the presentation of type 1 diabetes in five risk groups based on autoantibody combinations. For the first time, we were able to include for comparison children with very low risk of progression to type 1 diabetes. The Finnish Diabetes Prediction and Prevention birth cohort study screened newborn infants for HLA susceptibility to type 1 diabetes since 1994. Those carrying a risk genotype were prospectively followed up with islet autoantibody testing. Glucose parameters were obtained starting from the time of seroconversion. By 31 August 2014, 1162 children had developed at least one islet autoantibody and were included in the current study. Type 1 diabetes was diagnosed in 335 children (progressors). In the non-progressor groups, 207 developed multiple (≥2) biochemical islet autoantibodies, 229 a single biochemical autoantibody, 370 ICA only, and 64 transient autoantibodies. Children were divided into five risk groups. Glucose metabolism was evaluated. We observed lower HbA1c values in early follow-up 4.5 to 6.0 years before diagnosis in the progressors when compared to the same time in children with a single biochemical autoantibody or low-risk (ICA only and transient) participants, who did not progress to clinical type 1 diabetes. However, no such differences were observed in OGTTs or random plasma glucose. The variation was minimal in glucose values in the low-risk groups. We report the possibility of early alteration in glucose metabolism in future progressors. This could suggest early defects in multiple glucose-regulating hormones.


Hip fracture in Hong Kong and Britain.
Hip fractures in elderly people are an important public health problem in many Western populations. Little is known of their epidemiology in Oriental populations. A comparative study of hip fracture incidence in Hong Kong and Southampton showed similar incidence rates by age and sex. In the last two decades hip fracture incidence in Hong Kong has increased, particularly among men. This can be partly explained by reduction in the levels of activity among the elderly Chinese population.


The Social Dimension of Adolescent Health: Challenges for Paediatric Research and Practice.
Innumerable studies have shown that socioeconomic inequalities have a serious impact on mortality and morbidity. Disease and premature mortality are thus embodied expressions of the conditions under which we live and work. The increasing discussion of adolescence as an independent stage of life has generated a growth of interest in the social determinants of young people's health. The review outlines that a large part of the prevailing diseases in adolescence are strongly influenced by social, economic and political determinants and analyses which determinants and mechanisms are responsible that social inequalities get under the skin and cause adverse health. Paediatrics offers a central interdisciplinary link between the natural and social sciences to better understand how we biologically incorporate our lived experience and thus create social patterns of health and illness not only in societies but also between societies. Such research could contribute to further sensitise clinical and therapeutic practice on the social determinants of health and to integrate them in daily routine processes, such as anamnesis and therapy.


Negative regulation of erythropoiesis by caspase-mediated cleavage of GATA-1.
The production of red blood cells follows the sequential formation of proerythroblasts and basophilic, polychromatophilic and orthochromatic erythroblasts, and is promoted by the hormone erythropoietin (Epo) in response to tissue hypoxia. However, little is known about the negative regulation of this process. Death receptors are a family of surface molecules that trigger caspase activation and apoptosis in a variety of cell types. Here we show that immature erythroid cells express several death receptors whose ligands are produced by mature erythroblasts. Exposure of erythroid progenitors to mature erythroblasts or death-receptor ligands resulted in caspase-mediated degradation of the transcription factor GATA-1, which is associated with impaired erythroblast development. Expression of a caspase-resistant GATA-1 mutant, but not of the wild-type gene, completely restored erythroid expansion and differentiation following the triggering of death receptors, indicating that there is regulatory feedback between mature and immature erythroblasts through caspase-mediated cleavage of GATA-1. Similarly, erythropoiesis blockade following Epo deprivation was largely prevented by the expression of caspase-inhibitory proteins or caspase-resistant GATA-1 in erythroid progenitors. Caspase-mediated cleavage of GATA-1 may therefore represent an important negative control mechanism in erythropoiesis.


Intrapulpal temperature during direct fabrication of provisional restorations.
Measurements were made of the heat transferred to the pulp by different types of resins, different volumes of resin, and different types of matrices commonly used in the direct fabrication of extracoronal provisional restorations. A mandibular model was fabricated using extracted teeth that were prepared for extracoronal restorations. Provisional restorations were fabricated for the prepared teeth using conventional direct techniques, and the intrapulpal temperature rise was recorded. The temperature rise ranged from a low of 0.42 degrees C to a high of 7.21 degrees C. Of the three resins tested, the poly(methyl methacrylate) material induced the greatest temperature increase. The vacuum-formed matrix allowed a greater temperature increase than either of the impression material matrices.


Socio-economic benefits from protected areas in southeastern Australia.
International case studies of protected area performance increasingly report that conservation and socio-economic outcomes are interdependent. Effective conservation requires support and cooperation from local governments and communities, which in turn requires that protected areas contribute to the economic well-being of the communities in which they are sited. Despite increasing recognition of their importance, robust studies that document the socio-economic impacts of protected areas are rare, especially in the developed world context. We proposed 3 potential pathways through which protected areas might benefit local communities in the developed world: the improved local housing value, local business stimulus, and increased local funding pathways. We examined these pathways by undertaking a statistical longitudinal analysis of 110 regional and rural communities covering an area of approximately 600,000 km(2) in southeastern Australia. We compared trends in 10 socio-economic indicators describing employment, income, housing, business development and local government revenue from 2000 to 2010. New protected areas acquisitions led to an increased number of new dwelling approvals and associated developer contributions, increased local business numbers, and increased local government revenue from user-pays services and grants. Longer-term effects of established protected areas included increased local council revenue from a variety of sources. Our findings provide support for each of our 3 proposed benefit pathways and contribute new insights into the cycling of benefits from protected areas through the economy over time. The business and legislative models in our study are typical of those operating in many other developed countries; thus, the benefit pathways reported in our study are likely to be generalizable. By identifying and communicating socio-economic benefits from terrestrial protected areas in a developed world context, our findings represent an important step in securing local support and ongoing high-level protection for key components of the world's biodiversity.


Cohesin-dockerin code in cellulosomal dual binding modes and its allosteric regulation by proline isomerization.
Cellulose is the most abundant organic molecule on Earth and represents a renewable and practically everlasting feedstock for the production of biofuels and chemicals. Self-assembled owing to the high-affinity cohesin-dockerin interaction, cellulosomes are huge multi-enzyme complexes with unmatched efficiency in the degradation of recalcitrant lignocellulosic substrates. The recruitment of diverse dockerin-borne enzymes into a multicohesin protein scaffold dictates the three-dimensional layout of the complex, and interestingly two alternative binding modes have been proposed. Using single-molecule fluorescence resonance energy transfer and molecular simulations on a range of cohesin-dockerin pairs, we directly detect varying distributions between these binding modes that follow a built-in cohesin-dockerin code. Surprisingly, we uncover a prolyl isomerase-modulated allosteric control mechanism, mediated by the isomerization state of a single proline residue, which regulates the distribution and kinetics of binding modes. Overall, our data provide a novel mechanistic understanding of the structural plasticity and dynamics of cellulosomes.


Methylene Blue for Bronchopleural Fistula Localization.
A bronchopleural fistula (BPF) is a communication between the pleural space and the bronchial tree. BPFs are challenging to diagnose and are associated with a high morbidity and mortality. Sequential balloon occlusion is commonly used for localization of a BPF. We describe our experience with 4 cases of successful localization of the BPF by instillation of methylene blue into the pleural space through a pigtail catheter, with simultaneous bronchoscopic visualization of dye in the tracheobronchial tree. Two patients were treated with endobronchial valves and 3 had a surgical thoracic muscle flap placed.


Exploring the views of nurse prescribing among Macmillan nurses.
Nurse prescribing is a key government initiative which aims to enhance patient care. The aim of this study was to investigate the prescribing practice of specialist nurses working in cancer and palliative care and to explore the benefits of and barriers to uptake of nurse prescribing training. A national postal survey of 2252 Macmillan nurses was undertaken using a structured questionnaire with open-ended questions. Data were analyzed using thematic content analysis for 1575 respondents (70% response rate). Only 13% (203) had undergone prescribing training and of these 105 provided responses to the open questions concerning training and of the 87% (1372) who had not undergone the training, 423 provided details on barriers to nurse prescribing training. The data presented in this paper draw from this data. The findings indicate that those who prescribed gave the prospect of improving care as the main reason for undertaking nurse prescribing training. The main reasons why these specialist nurses did not undertake training were: resource issues particularly with respect to backfill while training, lack of medical support and mentorship, concerns about the relevance of prescribing as a nursing role and prioritizing other courses. If nurse prescribing is to be more widely available for cancer and palliative care patients it is important in both primary and secondary care to address the resource and support issues.


[Mortality at an internal medicine department of a tertiary hospital].
The clinical and epidemiological variables as well as conduct parameters, attributable to death cases of the 1,052 patients cared for during one year in an Internal Medicine Department are analyzed. The mortality rate was 11.6%. We may point out in our results the elevated mean age (75.5 +/- 11.2 years) and the high incidence of repeated admissions (46%) among death cases. The mean and median hospital stay values were lower in the patients who died than in those that survived. The more frequent basic causes of death were stroke (27%), chronic obstructive pulmonary disease (15%) and neoplasias (14%). When analyzing the different conditions within one patient, cardiovascular diseases were most often found amongst those who died (73%). Gastrointestinal and infectious diseases were on the contrary associated to a low mortality rate.


MR angiography-planned prostatic artery embolization for benign prostatic hyperplasia: single-center retrospective study in 56 patients.
We aimed to evaluate the advantages of magnetic resonance angiography (MRA)-planned prostatic artery embolization (PAE) for benign prostatic hyperplasia (BPH). In this retrospective study, MRAs of 56 patients (mean age, 67.23±7.73 years; age range, 47-82 years) who underwent PAE between 2017 and 2018 were evaluated. For inclusion, full information about procedure time and radiation values must have been available. To identify prostatic artery (PA) origin, three-dimensional MRA reconstruction with maximum intensity projection was conducted in every patient. In total, 33 patients completed clinical and imaging follow-up and were included in clinical evaluation. There were 131 PAs with a second PA in 19 pelvic sides. PA origin was correctly identified via MRA in 108 of 131 PAs (82.44%). In patients in which MRA allowed a PA analysis, a significant reduction of the fluoroscopy time (-27.0%, p = 0.028) and of the dose area product (-38.0%, p = 0.003) was detected versus those with no PA analysis prior to PAE. Intervention time was reduced by 13.2%, (p = 0.25). Mean fluoroscopy time was 30.1 min, mean dose area product 27,749 µGy•m2, and mean entrance dose 1553 mGy. Technical success was achieved in all 56 patients (100.0%); all patients were embolized on both pelvic sides. The evaluated data documented a significant reduction in IPSS (p < 0.001; mean 9.67 points). MRA prior to PAE allowed the identification of PA in 82.44% of the cases. MRA-planned PAE is an effective treatment for patients with BPH.


Ozone exposure upregulates the expression of host susceptibility protein TMPRSS2 to SARS-CoV-2.
SARS-CoV-2, a novel coronavirus and an etiologic agent for the current global health emergency, causes acute infection of the respiratory tract leading to severe disease and significant mortality. Ever since the start of SARS-CoV-2, also known as the COVID-19 pandemic, countless uncertainties have been revolving around the pathogenesis and epidemiology of the SARS-CoV-2 infection. While air pollution has been shown to be strongly correlated to increased SARS-CoV-2 morbidity and mortality, whether environmental pollutants such as ground-level ozone affects the susceptibility of individuals to SARS-CoV-2 is not yet established. To investigate the impact of ozone inhalation on the expression levels of signatures associated with host susceptibility to SARS-CoV-2, we analyzed lung tissues collected from mice that were sub-chronically exposed to air or 0.8 ppm ozone for three weeks (4 h/night, 5 nights/week), and analyzed the expression of signatures associated with host susceptibility to SARS-CoV-2. SARS-CoV-2 entry into the host cells is dependent on the binding of the virus to the host cellular receptor, angiotensin-converting enzyme (ACE2), and its subsequent proteolytic priming by the host-derived protease, transmembrane protease serine 2 (TMPRSS2). The Ace2 transcripts were significantly elevated in the parenchyma, but not in the extrapulmonary airways and alveolar macrophages, from ozone-exposed mice. The TMPRSS2 protein and Tmprss2 transcripts were significantly elevated in the extrapulmonary airways, parenchyma, and alveolar macrophages from ozone-exposed mice. A significant proportion of additional known SARS-CoV-2 host susceptibility genes were upregulated in alveolar macrophages and parenchyma from ozone-exposed mice. Our data indicate that the unhealthy levels of ozone in the environment may predispose individuals to severe SARS-CoV-2 infection. Given the severity of this pandemic and the challenges associated with direct testing of host-environment interactions in clinical settings, we believe that this ozone exposure-based study informs the scientific community of the potentially detrimental effects of the ambient ozone levels in determining the host susceptibility to SARS-CoV-2.


Integrating a multimode design into a national random-digit-dialed telephone survey.
The Behavioral Risk Factor Surveillance System (BRFSS) was originally conducted by using a landline telephone survey mode of data collection. To meet challenges of random-digit-dial (RDD) surveys and to ensure data quality and validity, BRFSS is integrating multiple modes of data collection to enhance validity. The survey of adults who use only cellular telephones is now conducted in parallel with ongoing, monthly landline telephone BRFSS data collection, and a mail follow-up survey is being implemented to increase response rates and to assess nonresponse bias. A pilot study in which respondents' physical measurements are taken is being conducted to assess the feasibility of collecting these data for a subsample of adults in 2 states. Physical measures would allow for the adjustment of key self-reported risk factor and health condition estimates and improve the accuracy and usefulness of BRFSS data. This article provides an overview of these new modes of data collection.


Surface electromyography analysis in long-term recordings: application to head rest comfort in cars.
Analysis of long-term surface electromyographic (SEMG) signals has many applications in ergonomics when related to muscle fatigue. The present work proposes a set of processing methods reporting SEMG modifications during long-term driving tests in various situations (with or without head rest). A segmentation/classification algorithm allows signal splitting into homogeneous parts (postural activity and EMG bursts) and an efficient artefact suppression. Postural activity modifications are evaluated from time-varying amplitude probability density function (TAPDF) parameters. EMG burst analysis is achieved taking into account the relationships of these bursts with accelerometric events. This segmentation/classification procedure improves repeatability but does not significantly modify the overall results obtained before segmentation, as far as the analysis of head rest influence is concerned.


Transplantation of embryonic stem cells improves nerve repair and functional recovery after severe sciatic nerve axotomy in rats.
Extensive research has focused on transplantation of pluripotent stem cells for the treatment of central nervous system disorders, the therapeutic potential of stem cell therapy for injured peripheral nerves is largely unknown. We used a rat sciatic nerve transection model to test the ability of implanted embryonic stem (ES) cell-derived neural progenitor cells (ES-NPCs) in promoting repair of a severely injured peripheral nerve. Mouse ES cells were neurally induced in vitro; enhanced expression and/or secretion of growth factors were detected in differentiating ES cells. One hour after removal of a 1-cm segment of the left sciatic nerve, ES-NPCs were implanted into the gap between the nerve stumps with the surrounding epineurium as a natural conduit. The transplantation resulted in substantial axonal regrowth and nerve repair, which were not seen in culture medium controls. One to 3 months after axotomy, co-immunostaining with the mouse neural cell membrane specific antibody M2/M6 and the Schwann cell marker S100 suggested that transplanted ES-NPCs had survived and differentiated into myelinating cells. Regenerated axons were myelinated and showed a uniform connection between proximal and distal stumps. Nerve stumps had near normal diameter with longitudinally oriented, densely packed Schwann cell-like phenotype. Fluoro-Gold retrogradely labeled neurons were found in the spinal cord (T12-13) and DRG (L4-L6), suggesting reconnection of axons across the transection. Electrophysiological recordings showed functional activity recovered across the injury gap. These data suggest that transplanted neurally induced ES cells differentiate into myelin-forming cells and provide a potential therapy for severely injured peripheral nerves.


The autonomous growth of human papillomavirus type 16-immortalized keratinocytes is related to the endothelin-1 autocrine loop.
Some human papillomaviruses (HPVs) such as HPV type 16 (HPV16) and HPV18 are involved in cervical carcinoma, and they can immortalize and transform keratinocytes. Endothelin-1 (ET-1) is produced in keratinocytes and has been shown to act through ETA receptors as an autocrine growth factor for keratinocytes. This study examines whether HPV16 alters the ET-1-mediated autocrine loop in human keratinocytes, providing a selective growth advantage for transformed cells. ET-1 is released in similar amounts from normal and HPV-transfected keratinocytes. All HPV-transfected cell lines express high-affinity ETA receptors. A two-fold increase in ET-1 binding sites is present in HPV16-immortalized keratinocytes, and this effect seems to be linked to the overexpression of mRNA for this receptor rather than to differences in the surface/internalized ratio of the receptors. ET-1 induces significant increases in [3H]thymidine incorporation and cell proliferation. Furthermore, HPV-transfected keratinocytes can proliferate in the absence of any growth factor added to the growth medium, and the ETA receptor antagonist BQ123 prevents this proliferation. These data suggest a new mechanism in the growth control of HPV-transformed cells mediated by the upregulation of ET-1 autocrine loop.


Naturally acquired attaching and effacing Escherichia coli in sheep.
In a series of experiments involving the inoculation of sheep with Escherichia coli O157:H7, and subsequent detailed histopathological examination of the intestinal mucosa, attaching-effacing (AE) lesions formed by elements of the natural flora were observed in 18% of animals. These incidental AE lesions typically were small and sparse, and were not associated with clinical disease. It was possible to identify further some of the lesional bacteria, revealing that E. coli O115 had formed lesions in one of the seven affected animals, and similarly E. coli O26 had formed some of the lesions in another. As AE strains, source flocks, housing and feed sources were diverse, a common source of lesion-forming bacteria appears to be unlikely. It is postulated that subclinical AE lesions are a mechanism of persistence of AE bacteria in sheep.


Novel mutations in congenital factor XII deficiency.
Several mutations in factor XII have been reported in patients with factor XII deficiency. Here, we described three mutations in the F12 gene (c. 6635G more than A (p. G259E), c. 6658G more than C (p. R267G) and c. 8489G more than A (p. E521K)) of five patients with congenital FXII deficiency. Among these, two were heterozygous mutations. All five patients had prolonged activated partial thromboplastin time, as well as markedly decreased FXII activity and antigen levels. In vitro studies in transiently transfected HEK 293T cells demonstrated that these mutations significantly lowered the FXII levels in the culture media, but had no impact on transcription. Further protein degradation inhibition experiments with various inhibitors suggested that the three mutants were degraded intracellularly through the proteasome pathway in the pre-Golgi compartment. Moreover, G259E and R267G mutations exhibited dominant negative effects, consistent with the phenotypes observed in the heterozygous carriers. Such dominant negative effects were not due to the dimerization of FXII. Our findings suggest that the three mutations in the F12 gene are the causing reasons for the cross-reactive material-negative FXII deficiencies.


Biochemical pharmacology and DNA-drug interactions by Cl-958, a new antitumor intercalator derived from a series of substituted 2H-[1]benzothiopyrano[4,3,2-cd]indazoles.
Cl-958, PD 121373, and PD 114595 belong to a new class of DNA complexers, substituted 2H-[1]benzothiopyrano[4,3,2-cd] indazoles, and are being further developed as antitumor drugs based on their curative properties against murine solid tumor models. The biochemical effects of these drugs on L1210 leukemia cells and their interaction with DNA were studied and compared to clinically used intercalators. The benzothiopyranoindazoles bound to DNA with a relatively high affinity, having intrinsic association constants of between 3 and 4 x 10(5) M-1. Based on viscosity measurements, the mode of DNA binding appears to be through intercalation. Unwinding angles were calculated to be approximately 18 degrees. The benzothiopyranoindazoles were potent inhibitors of nucleic acid synthesis, reducing both DNA and RNA synthesis to the same extent at similar concentrations. Like other known intercalators, these compounds produced DNA single- and double-strand breaks in a time- and concentration-dependent manner in L1210 cells. Between one and two DNA strand breaks were formed per protein-strand crosslink. Repair of these DNA lesions after the drug was removed from the cells was either very slow or did not occur at all for at least 2 hr. Finally, since the high incidence of cardiotoxicity associated with the administration of anthracyclines has been related to the formation of reactive oxygen species, the ability of the benzothiopyranoindazoles to augment superoxide dismutase-sensitive oxygen consumption was observed in a rat liver microsomal system. These compounds produced less than 5% of the activity in this assay that doxorubicin produced.


Extracellular Matrix Proteomics: The mdx-4cv Mouse Diaphragm as a Surrogate for Studying Myofibrosis in Dystrophinopathy.
The progressive degeneration of the skeletal musculature in Duchenne muscular dystrophy is accompanied by reactive myofibrosis, fat substitution, and chronic inflammation. Fibrotic changes and reduced tissue elasticity correlate with the loss in motor function in this X-chromosomal disorder. Thus, although dystrophinopathies are due to primary abnormalities in the DMD gene causing the almost-complete absence of the cytoskeletal Dp427-M isoform of dystrophin in voluntary muscles, the excessive accumulation of extracellular matrix proteins presents a key histopathological hallmark of muscular dystrophy. Animal model research has been instrumental in the characterization of dystrophic muscles and has contributed to a better understanding of the complex pathogenesis of dystrophinopathies, the discovery of new disease biomarkers, and the testing of novel therapeutic strategies. In this article, we review how mass-spectrometry-based proteomics can be used to study changes in key components of the endomysium, perimysium, and epimysium, such as collagens, proteoglycans, matricellular proteins, and adhesion receptors. The mdx-4cv mouse diaphragm displays severe myofibrosis, making it an ideal model system for large-scale surveys of systematic alterations in the matrisome of dystrophic fibers. Novel biomarkers of myofibrosis can now be tested for their appropriateness in the preclinical and clinical setting as diagnostic, pharmacodynamic, prognostic, and/or therapeutic monitoring indicators.


Stressful life events and transitions in problematic alcohol use involvement among US adults.
We investigated the impact of stressful life events (SLEs) for males and females on transitions in problematic alcohol involvement, both progression and recovery, over a 3-year interval. Participants of both Wave 1 (2001-2002) and Wave 2 (2004-2005) of the National Epidemiologic Survey on Alcohol and Related Conditions (NESARC) were stratified by sex (14,233 males and 19,550 females). Latent transition analysis estimated the impact of experiencing ≥3 SLE in the year preceding the Wave 1 interview on the probability of transitioning between three empirically-derived stages of alcohol involvement (patterns of alcohol use disorder [AUD] symptoms), across waves. Propensity score methods adjusted for confounding. For males, three or more SLEs were associated with progression from the moderate to the severe problem stage (odds ratio [OR] = 2.23, 95% confidence interval [CI] = 1.17, 4.26). Among those in the severe problem stage, SLEs negatively impacted recovery regardless of sex. Employment/Financial SLEs were associated with a higher odds of transition from the moderate to the no problem stage (OR = 1.60, 95% CI = 1.03, 2.46) and lower odds of transitions from the severe to the moderate problem stage (OR = 0.40, 95% CI = 0.16, 0.99) among males, and from the severe to the no problem stage (OR = 0.26, 95% CI = 0.07, 0.88) among females. Stressful life events appear to affect transitions in alcohol involvement over time among those who already have alcohol problems, rather than impacting a transition among those without AUD problems.


A cryptic targeting signal induces isoform-specific localization of p46Shc to mitochondria.
The human Src homology and collagen (Shc) gene encodes three protein isoforms of 46, 52, and 66 kDa that belong to a family of molecular adapters involved in several signal transduction pathways. Recently, the 66-kDa isoform has been shown to play a central role in controlling reactive oxygen species metabolism and life span in mammals. Despite the large amount of information available on the biology and biochemistry of Shc proteins, very little is known regarding the regulation of their subcellular localization. Here we demonstrate the specific and selective localization of p46Shc to the mitochondrial matrix. Through deletion mapping experiments, we show that targeting of p46Shc to mitochondria is mediated by its first 32 amino acids, which behave as a bona fide mitochondrial targeting sequence. We further demonstrate that the N-terminal location of the signal peptide is critical for its function. This accounts for the observation that p52Shc and p66Shc, containing the same sequence but more internally located, display a remarkably different subcellular localization. These findings indicate that p46Shc may exert a non-redundant biological function in signal transduction pathways involving mitochondria.


Relations among training volume, body weight, and profile of mood states for elite judoka during a competitive period.
The present purpose was to evaluate changes in 10 elite judokas' mood state (5 men, 5 women) during the 7-wk. competitive period of the season. Quantitative monitoring of the training volume and body-weight monitoring were recorded, and an adapted Profile of Mood States was administered weekly, including competition days. Analysis showed that following a significant decrease in the specific training load volume, there were no significant changes in judokas' ratings on the Profile of Mood States; however, there was a decrease of 3 or 4% of the initial body weight during the precompetition weeks, which significantly affected mood state during competition, indicated by an increase in rated Tension and Vigor and a decrease in rated Fatigue.


Anti-tumour effects of a specific anti-ADAM17 antibody in an ovarian cancer model in vivo.
ADAM 17 (TNF-α converting enzyme, TACE) is a potential target for cancer therapy, but the small molecule inhibitors reported to date are not specific to this ADAM family member. This membrane-bound metalloproteinase is responsible for ectodomain shedding of pathologically significant substrates including TNF-α and EGFR ligands. The aim of this study was to evaluate the pharmacokinetics, pharmacodynamics and anti-tumour efficacy of the first specific inhibitor, an anti-human ADAM17 IgG antibody, clone D1(A12). We used intraperitoneal xenografts of the human ovarian cancer cell line IGROV1-Luc in Balb/c nude mice, chosen because it was previously reported that growth of these xenografts is inhibited by knock-down of TNF-α. In vitro, 200 nM D1(A12) inhibited shedding of ADAM17 substrates TNF-α, TNFR1-α, TGF-α, amphiregulin (AREG), HB-EGF and IL-6Rα, from IGROV1-Luc cells, (4.7 nM IC(50) for TNF-α shedding). In IGROV1-Luc xenografts in vivo, D1(A12) IgG showed pharmacokinetic properties suitable for efficacy studies, with a single i.p. dose of 10 mg/kg D1(A12) sufficient to maintain IgG plasma and ascites fluid concentrations above 100 nM for more than 7 days. The plasma half life was 8.6 days. Next, an efficacy study was performed, dosing D1(A12) or anti-human TNF-α antibody infliximab at 10 mg/kg q7d, quantifying IGROV1-Luc tumour burden by bioluminescence. D1(A12) IgG showed a significant reduction in tumour growth (p = 0.005), 56% of vehicle control. Surprisingly, D1(A12) did not reduce the concentration of circulating human TNF-α, suggesting that another enzyme may compensate for inhibition of ADAM17 in vivo (but not in vitro). However, D1(A12) did show clear pharmacodynamic effects in the mice, with significant inhibition of shedding from tumour of ADAM17 substrates TNFR1-α, AREG, and TGF-α (4-15-fold reductions, p<0.0001 for all three). Thus, D1(A12) has anti-ADAM17 activity in vivo, inhibits shedding of EGFR ligands and has potential for use in EGF ligand-dependent tumours.


Posttreatment occlusal variability among angle Class I nonextraction patients.
To test the hypothesis that variability among posttreatment Angle Class I, nonextraction patients is not related to patient and treatment factors. One hundred thirty-eight subjects were randomly selected from the archives of the Department of Orthodontics, St Louis University. Patient factors evaluated included age, sex, mandibular plane angle, and ANB angle; treatment factors included active treatment time and supervising orthodontist. Objective Grading System (OGS) scores for six of the criteria (excluding interproximal contacts and root angulations) and the anterior Bolton ratio were measured on the posttreatment study casts. The partial mean overall OGS score was 24.9 +/- 8.0. Occlusal contact was the most important component contributing to the overall score, followed by alignment. Variation in the OGS scores was explained by sex, pretreatment mandibular plane and ANB angles, the posttreatment anterior Bolton ratio, and treatment duration. The partial overall OGS scores increased by approximately one point for every 4 degrees increase in the mandibular plane angle and nearly one point for every 3 additional months of treatment. Approximately 16% and 15% of the variation in alignment and buccolingual inclination, respectively, was due to the treating orthodontist. Posttreatment occlusal variability among Class I nonextraction patients can be partially explained by patient- and treatment-related factors.


Serum 90K/Mac-2 binding protein (Mac-2BP) as a response predictor to peginterferon and ribavirin combined treatment in HCV chronic patients.
90K/Mac-2BP glycoprotein is involved in the immune defense against a variety of neoplasms and viral infections, modulating the activity of several effectors such as natural killer cells. Quite interestingly, 90K/Mac-2BP is associated to a poor response to interferon (IFN) alpha in hepatitis C virus (HCV) infected patients. Here, in 70 consecutive HCV chronic patients, we have evaluated 90K basal levels as a response predictor to combined therapy with Peginterferon and Ribavirin. We have found higher 90K levels in genotype 1/4 than in genotype 2/3 (p = 0.006) and in 62.5% of non-responders than in 20% of responders (p < 0.001). Genotype 1/4, higher 90K and gamma glutamyl transferase (gammaGT) levels resulted independently associated to a status of refractoriness to therapy. Consequently, evaluation of 90K serum levels seems to be a promising useful marker of response to combined therapy in HCV disease.


Clinical efficacy and prognostic indicators for lower limb pedalling exercise early after stroke: study protocol for a pilot randomised controlled trial.
It is known that repetitive, skilled, functional movement is beneficial in driving functional reorganisation of the brain early after stroke. This study will investigate a) whether pedalling an upright, static exercise cycle, to provide such beneficial activity, will enhance recovery and b) which stroke survivors might be able to participate in pedalling. Participants (n = 24) will be up to 30 days since stroke onset, with unilateral weakness and unable to walk without assistance. This study will use a modified exercise bicycle fitted with a UniCam crank. All participants will give informed consent, then undergo baseline measurements, and then attempt to pedal. Those able to pedal will be entered into a single-centre, observer-blinded randomised controlled trial (RCT). All participants will receive routine rehabilitation. The experimental group will, in addition, pedal daily for up to ten minutes, for up to ten working days.Prognostic indicators, measured at baseline, will be: site of stroke lesion, trunk control, ability to ambulate, and severity of lower limb paresis.The primary outcome for the RCT is ability to voluntarily contract paretic lower limb muscle, measured by the Motricity Index. Secondary outcomes include ability to ambulate and timing of onset and offset of activity in antagonist muscle groups during pedalling, measured by EMG. This protocol is for a trial of a novel therapy intervention. Findings will establish whether there is sufficient evidence of benefit to justify proceeding with further research into clinical efficacy of upright pedalling exercise early after stroke. Information on potential prognostic indicators will suggest which stroke survivors could benefit from the intervention. ISRCTN: ISRCTN45392701.


Hepatitis B virus X protein stimulates viral genome replication via a DDB1-dependent pathway distinct from that leading to cell death.
The hepatitis B virus (HBV) X protein (HBx) is essential for virus infection and has been implicated in the development of liver cancer associated with chronic infection. HBx can interact with a number of cellular proteins, and in cell culture, it exhibits pleiotropic activities, among which is its ability to interfere with cell viability and stimulate HBV replication. Previous work has demonstrated that HBx affects cell viability by a mechanism that requires its binding to DDB1, a highly conserved protein implicated in DNA repair and cell cycle regulation. We now show that an interaction with DDB1 is also needed for HBx to stimulate HBV genome replication. Thus, HBx point mutants defective for DDB1 binding fail to complement the low level of replication of an HBx-deficient HBV genome when provided in trans, and one such mutant regains activity when directly fused to DDB1. Furthermore, DDB1 depletion by RNA interference specifically compromises replication of wild-type HBV, indicating that HBx produced from the viral genome also functions in a DDB1-dependent fashion. We also show that HBx in association with DDB1 acts in the nucleus and stimulates HBV replication mainly by enhancing viral mRNA levels, regardless of whether the protein is expressed from the HBV genome itself or supplied in trans. Interestingly, whereas HBx induces cell death in both HepG2 and Huh-7 hepatoma cell lines, it enhances HBV replication only in HepG2 cells, suggesting that the two activities involve distinct DDB1-dependent pathways.


Dynamics of metabolic activities and gene expression in the Roseobacter clade bacterium Phaeobacter sp. strain MED193 during growth with thiosulfate.
Metagenomic analyses of surface seawater reveal that genes for sulfur oxidation are widespread in bacterioplankton communities. However, little is known about the metabolic processes used to exploit the energy potentially gained from inorganic sulfur oxidation in oxic seawater. We therefore studied the sox gene system containing Roseobacter clade isolate Phaeobacter sp. strain MED193 in acetate minimal medium with and without thiosulfate. The addition of thiosulfate enhanced the bacterial growth yields up to 40% in this strain. Concomitantly, soxB and soxY gene expression increased about 8-fold with thiosulfate and remained 11-fold higher than that in controls through stationary phase. At stationary phase, thiosulfate stimulated protein synthesis and anaplerotic CO2 fixation rates up to 5- and 35-fold, respectively. Several genes involved in anaplerotic CO2 fixation (i.e., pyruvate carboxylase, propionyl coenzyme A [CoA], and crotonyl-CoA carboxylase) were highly expressed during active growth, coinciding with high CO2 fixation rates. The high expression of key genes in the ethylmalonyl-CoA pathway suggests that this is an important pathway for the utilization of two-carbon compounds in Phaeobacter sp. MED193. Overall, our findings imply that Roseobacter clade bacteria carrying sox genes can use their lithotrophic potential to gain additional energy from sulfur oxidation for both increasing their growth capacity and improving their long-term survival.


A novel mutant variant of the CYP2D6 gene (CYP2D6*17) common in a black African population: association with diminished debrisoquine hydroxylase activity.
1. The debrisoquine hydroxylase (CYP2D6) is polymorphically distributed. Not only are there differences in the proportions of extensive metabolisers to poor metabolisers in various ethnic groups, but there are also pronounced variations in the metabolic capacity among those classified as extensive metabolisers. 2. The mean debrisoquine metabolic ratio of Caucasian extensive metabolisers is lower than that for a number of African populations. In the present study, we have searched for novel CYP2D6 mutations to explain the diminished enzyme activity in African populations. 3. Three Zimbabwean Shona subjects with EM phenotypes (metabolic ratios for debrisoquine of 0.4, 1.5 and 10.5 respectively) were selected and the open reading frame of the CYP2D6 gene of each was sequenced. 4. The subject with metabolic ratio of 10.5 was found to be homozygous for an allele with a nucleotide exchange in exon 2, 1111C-->T causing a 107Thr-->Ile amino acid exchange in a conserved region of the enzyme. In addition, he was homozygous for the 2938C-->T and 4268G-->C mutations causing 296Arg-->Ser and 486Ser-->Thr amino acid substitution found in the CYP2D6*2 allele. 5. Seventy-six Zimbabwean Shona subjects were subsequently genotyped for the 1111C-->T mutation and for the intron 1 gene conversion present in the CYP2D6*2 gene. The 1111C-->T mutation was found at an allele frequency of 34% and was only present in alleles carrying the gene conversion in intron 1 indicative for the CYP2D6*2 gene. 6. This allele (CYP2D6*17), containing the 1111C-->T, 2938C-->T and 4268G-->C mutations, was found to be strongly associated with lower capacity for debrisoquine hydroxylation. We therefore postulate that the CYP2D6*17 allele might contribute to the molecular basis of the previously established diminished debrisoquine hydroxylase activity in African Bantu populations.


Single Circuit in V1 Capable of Switching Contexts During Movement Using an Inhibitory Population as a Switch.
As animals adapt to their environments, their brains are tasked with processing stimuli in different sensory contexts. Whether these computations are context dependent or independent, they are all implemented in the same neural tissue. A crucial question is what neural architectures can respond flexibly to a range of stimulus conditions and switch between them. This is a particular case of flexible architecture that permits multiple related computations within a single circuit. Here, we address this question in the specific case of the visual system circuitry, focusing on context integration, defined as the integration of feedforward and surround information across visual space. We show that a biologically inspired microcircuit with multiple inhibitory cell types can switch between visual processing of the static context and the moving context. In our model, the VIP population acts as the switch and modulates the visual circuit through a disinhibitory motif. Moreover, the VIP population is efficient, requiring only a relatively small number of neurons to switch contexts. This circuit eliminates noise in videos by using appropriate lateral connections for contextual spatiotemporal surround modulation, having superior denoising performance compared to circuits where only one context is learned. Our findings shed light on a minimally complex architecture that is capable of switching between two naturalistic contexts using few switching units.


Making Sense of Memorable Messages About Infertility: Examining Message Valence by Theme and Sender.
Fertility problems, often called infertility, have been defined as the inability to conceive or maintain pregnancy throughout one year of trying (World Health Organization, 2020). Because fertility problems can present unique medical, emotional, relational, and identity challenges, they are often difficult to talk about, and even well-intentioned messages can be perceived negatively. This study uses Communicated Sense-Making (CSM; Kellas & Kranstuber Horstman, 2015), particularly its mechanism of memorable messages, to explore what types of support-related messages people experiencing infertility find memorable. Results from semi-structured interviews (N = 54) indicate five supra-themes of memorable messages: (a) communicating solidarity; (b) attempting to minimize participants' stress; (c) communicating investment or interest in the patient's experience; (d) sharing expertise; and (e) absolving the patient of responsibility; we identify several sub-themes within each. We also explore patterns between message types, senders, and message valence: message themes were perceived as either positive, negative, or neutral based on the combination of sender and perceived intention. Theoretical and practical implications are discussed.


[Vapor condensate of exhaled air in evaluating the impaired metabolism of the bronchopulmanory system in nonspecific lung diseases].
The degree of metabolic rehabilitation of the bronchopulmonary system was evaluated in non-specific pulmonary diseases, like pneumonia or chronic obstructive bronchitis, by using the data of biochemical testing of the exhaled-air vapor condensate. Nine parameters were investigated, i.e. enzymes alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase, alkaline phosphatase (AP), gamma-glutamate amino-transpeptidase (GGT) as well as parameters of protein metabolism--common protein, seromucoid (SC), C-reactive protein and urea. AST, ALT, AP, GGT, SC and urea were acknowledged as the most informative parameters. The results are indicative of that the recovery of metabolic processes in the bronchopulmonary system was not completed.


The shirker's dilemma and the prospect of cooperation in large groups.
Cooperation usually becomes harder to sustain as groups become larger because incentives to shirk increase with the number of potential contributors to collective action. But is this always the case? Here we study a binary-action cooperative dilemma where a public good is provided as long as not more than a given number of players shirk from a costly cooperative task. We find that at the stable polymorphic equilibrium, which exists when the cost of cooperation is low enough, the probability of cooperating increases with group size and reaches a limit of one when the group size tends to infinity. Nevertheless, increasing the group size may increase or decrease the probability that the public good is provided at such an equilibrium, depending on the cost value. We also prove that the expected payoff to individuals at the stable polymorphic equilibrium (i.e., their fitness) decreases with group size. For low enough costs of cooperation, both the probability of provision of the public good and the expected payoff converge to positive values in the limit of large group sizes. However, we also find that the basin of attraction of the stable polymorphic equilibrium is a decreasing function of group size and shrinks to zero in the limit of very large groups. Overall, we demonstrate non-trivial comparative statics with respect to group size in an otherwise simple collective action problem.


Comparison of Modified Above-Knee and Conventional Surgery with the Stripping of the Great Saphenous Vein of Varicose Veins of the Lower Extremities: A Retrospective Study.
To compare the clinical effects of modified above-knee and conventional surgery with the stripping of the great saphenous vein of varicose veins of the lower extremities. Clinical data of patients with a varicose vein of the lower extremity from May 2016 to May 2018 were collected. A retrospective study was conducted on the patients receiving modified above-knee and conventional surgery with the great saphenous vein stripping. The baseline characteristics and long-term follow-up data were compared between the groups. There were no significant differences in baseline characteristics between the two groups (P > 0.05). The surgeries were successfully performed by the same group of surgeons under local anesthesia and neuraxial anesthesia. The hospital stay, operation time, intraoperative blood loss, total length, and number of incisions in the above-knee group were comparable to those in the conventional surgery group (P > 0.05). The incidence of saphenous nerve injury and subcutaneous hematoma in the above-knee group was lower than that in the conventional surgery group (P < 0.05). There were no significant differences in recurrent varicose vein incidences (P > 0.05). After surgery, the venous clinical severity score (VCSS) and chronic venous insufficiency questionnaire (CIVIQ-14) scores of both groups were higher than those before operation (P < 0.05). There was no significant difference in VCSS score or CIVIQ-14 scores between the two groups postoperation (P > 0.05). At 24 months after surgery, the above-knee group (71.8%) and conventional surgery group (73.2%) resulted in changes of at least two CEAP-C clinical classes lower than baseline, respectively. The modified above-knee technique can ensure clinical outcomes, reduce intraoperative blood loss and complication incidences, and shorten the operative time. This gives evidence that the modified above-knee technique is worthy of clinical application.


Cyclic Ketones as Future Fuels: Reactivity with OH Radicals.
For a sustainable energy future, research directions should orient toward exploring new fuels suitable for future advanced combustion engines to achieve better engine efficiency and significantly less harmful emissions. Cyclic ketones, among bio-derived fuels, are of significant interest to the combustion community for several reasons. As they possess high resistance to autoignition characteristics, they can potentially be attractive for fuel blending applications to increase engine efficiency and also to mitigate harmful emissions. Despite their importance, very few studies are rendered in understanding of the chemical kinetic behavior of cyclic ketones under engine-relevant conditions. In this work, we have conducted an experimental investigation for the reaction kinetics of OH radicals with cyclopentanone and cyclohexanone for the first time over a wide range of experimental conditions ( T = 900-1330 K and p ≈ 1.2 bar) in a shock tube. Reaction kinetics was followed by monitoring UV laser absorption of OH radicals near 306.7 nm. Our measured rate coefficients, with an overall uncertainty (2σ) of ±20%, can be expressed in Arrhenius form as (in units of cm3 molecule-1 s-1): k1(CPO+OH)=1.20×10-10exp(-2115KT) (902-1297 K); k2(CHO+OH)=2.11×10-10exp(-2268KT) (935-1331 K). Combining our measured data with the single low-temperature literature data, the following three-parameter Arrhenius expressions (in units of cm3 molecule-1 s-1) are obtained over a wider temperature range: k1(CPO + OH) = 1.07×10-13(T300K)3.20exp(1005.7KT) (298-1297 K); k2(CHO+OH)=3.12×10-13(T300K)2.78exp(897.5KT) (298-1331 K). Discrepancies between the theoretical and current experimental results are observed. Earlier theoretical works are found to overpredict our measured rate coefficients. Interestingly, these cyclic ketones exhibit similar reactivity behavior to that of their linear ketone counterparts over the experimental conditions of this work.


A Modern View of the Interstitial Space in Health and Disease.
Increases in the volume of the interstitial space are readily recognized clinically as interstitial edema formation in the loose connective tissue of skin, mucosa, and lung. However, the contents and the hydrostatic pressure of this interstitial fluid can be very difficult to determine even in experimental settings. These difficulties have long obscured what we are beginning to appreciate is a dynamic milieu that is subject to both intrinsic and extrinsic regulation. This review examines current concepts regarding regulation of interstitial volume, pressure, and flow and utilizes that background to address three major topics of interest that impact IV fluid administration. The first of these started with the discovery that excess dietary salt can be stored non-osmotically in the interstitial space with minimal impact on vascular volume and pressures. This led to the hypothesis that, along with the kidney, the interstitial space plays an active role in the long-term regulation of blood pressure. Second, it now appears that hypovolemic shock leads to systemic inflammatory response syndrome principally through the entry of digestive enzymes into the intestinal interstitial space and the subsequent progression of enzymes and inflammatory agents through the mesenteric lymphatic system to the general circulation. Lastly, current evidence strongly supports the non-intuitive view that the primary factor leading to inflammatory edema formation is a decrease in interstitial hydrostatic pressure that dramatically increases microvascular filtration.


Results of Anatomical Arthroscopic Repair of Anterior Talofibular Ligament in Chronic Lateral Ankle Instability Patients.
To assess the clinical and functional outcomes of all-inside arthroscopic anatomical repair of anterior talofibular ligament (ATFL) for management of chronic lateral ankle instability (CLAI) in a considerable number of patients during medium-term follow-up. A retrospective analytic study was performed on 100 patients with CLAI who presented between August 2015 and July 2020 (average age: 32.9 years; range: 16-54 years). All-inside arthroscopic ATFL direct repair was performed in all patients through 2 portals only with fixation using 2 knotless anchors. Associated intraarticular lesions were treated in the same procedure. Outcomes were assessed with pre- and postoperative visual analog scale (VAS), the ankle-hindfoot score of the American Orthopaedic Foot & Ankle Society (AOFAS), and the Karlsson Ankle Functional Score (KAFS). All patients were followed for 24-48 months. At the final follow-up, ankle pain had improved significantly. Both the ankle anterior drawer test and the ankle varus stress tests were negative. There was no loss of ankle range of motion compared with preoperative measures, and all patients returned to normal gait. The mean VAS score decreased to 0.39 ± 0.63, the AOFAS score increased to 95.17 ± 4.7, and the KAFS score increased to 95 ± 4.07. All the follow-up indexes significantly improved compared to those before surgery. At minimum 24-month follow-up, the all-inside arthroscopic ATFL repair used to treat CLAI was found to restore ankle stability and yield good clinical outcomes with a relatively low complication rate. Level III, retrospective cohort study.


QT interval and antidepressant use: a cross sectional study of electronic health records.
To quantify the impact of citalopram and other selective serotonin reuptake inhibitors on corrected QT interval (QTc), a marker of risk for ventricular arrhythmia, in a large and diverse clinical population. A cross sectional study using electrocardiographic, prescribing, and clinical data from electronic health records to explore the relation between antidepressant dose and QTc. Methadone, an opioid known to prolong QT, was included to demonstrate assay sensitivity. A large New England healthcare system comprising two academic medical centres and outpatient clinics. 38,397 adult patients with an electrocardiogram recorded after prescription of antidepressant or methadone between February 1990 and August 2011. Relation between antidepressant dose and QTc interval in linear regression, adjusting for potential clinical and demographic confounding variables. For a subset of patients, change in QTc after drug dose was also examined. Dose-response association with QTc prolongation was identified for citalopram (adjusted beta 0.10 (SE 0.04), P<0.01), escitalopram (adjusted beta 0.58 (0.15), P<0.001), and amitriptyline (adjusted beta 0.11 (0.03), P<0.001), but not for other antidepressants examined. An association with QTc shortening was identified for bupropion (adjusted beta 0.02 (0.01) P<0.05). Within-subject paired observations supported the QTc prolonging effect of citalopram (10 mg to 20 mg, mean QTc increase 7.8 (SE 3.6) ms, adjusted P<0.05; and 20 mg to 40 mg, mean QTc increase 10.3 (4.0) ms, adjusted P<0.01). This study confirmed a modest prolongation of QT interval with citalopram, and identified additional antidepressants with similar observed risk. Pharmacovigilance studies using electronic health record data may be a useful method of identifying potential risk associated with treatments.


A novel 3D integrated platform for the high-resolution study of cell migration plasticity.
Understanding the mechanisms of interstitial cancer migration is of great scientific and medical interest. Creating 3D platforms, conducive to optical microscopy and mimicking the physical parameters (in plane and out of plane) involved in interstitial migration, is a major step forward in this direction. Here, a novel approach is used to directly print free-form, 3D micropores on basal scaffolds containing microgratings optimized for contact guidance. The platforms so formed are validated by monitoring cancer cell migration and micropore penetration with high-resolution optical microscopy. The shapes, sizes and deformability of the micropores are controllable, paving the way to decipher their role in interstitial migration.


Effect of roaming transition states upon product branching in the thermal decomposition of CH3NO2.
The kinetics for the thermal unimolecular decomposition of CH3NO2 and its structural isomer CH3ONO have been investigated by statistical theory calculations based on the potential energy surface calculated at the UCCSD(T)/CBS and CASPT3(8, 8)/6-311+G(3df,2p) levels. Our results show that for the decomposition of CH3NO2 at pressures less than 2 Torr, isomerization to CH3ONO via the recently located roaming transition state is dominant in the entire temperature range studied, 400-3000 K. However, at higher pressures, the formation of the commonly assumed products, CH3 + NO2, becomes competitive and at pressures higher than 200 Torr the production of CH3 + NO2 is exclusive. The predicted rate constants for 760 Torr and the high-pressure limit with Ar as diluent in the temperature range 500-3000 K, producing solely CH3 + NO2, can be expressed respectively by kd(760)(CH3NO2) = 2.94 × 10(55)T(-12.6) exp(-35500/T) s(-1) and kd(∞)(CH3NO2) = 5.88 × 10(24)T(-2.35) exp(-31400/T) s(-1). In the low pressure limit, the decomposition reaction takes place exclusively via the roaming TS producing internally excited CH3ONO, giving rise to both CH3O + NO and CH2O + HNO with the second-order rate constant kd(0)(CH3NO2) = 1.17 × 10(31)T(-10.94) exp(-32400/T) cm(3) molecule(-1) s(-1). For CH3ONO decomposition, a new roaming transition state connecting to the CH2O + HNO products has been located, lying 6.8 kcal/mol below the well-known four-member ring tight transition state and 0.7 kcal/mol below CH3O + NO. The rate constants predicted by similar calculations give rise to the following expressions for the thermal decomposition of CH3ONO in He: kd(760)(CH3ONO) = 8.75 × 10(41)T(-8.97) exp(-22600/T) s(-1) and kd(∞)(CH3ONO) = 1.58 × 10(23)T(-2.18) exp(-21100/T) s(-1) in the temperature range 300-3000 K. These results are in very good agreement with available experimental data obtained under practical pressure conditions. The much different branching ratios for the formation of CH3O + NO and CH2O + HNO in the decomposition of both CH3NO2 and CH3ONO are also given in this work.


Ca2+, cAMP, and changes in myosin phosphorylation during contraction of smooth muscle.
Phosphorylation of myosin increases rapidly upon stimulation of an arterial smooth muscle. However, peak values are not maintained and phosphorylation declines, while active stress increases monotonically to a sustained steady state. The aim of this study was to determine the reason(s) for the transient change in myosin phosphorylation. Four hypotheses were considered: 1) reduced substrate, i.e., ATP depletion, 2) altered access of either the myosin kinase or phosphatase to the cross bridge, 3) reduced myosin kinase activity secondary to its phosphorylation by adenosine 3',5'-cyclic monophosphate-dependent protein kinase, and 4) reduced myoplasmic [Ca2+] during the contraction. Our results suggest that the most likely explanation is that there are two Ca2+-dependent regulatory processes: 1) myosin phosphorylation and 2) a second, unidentified site allowing stress maintenance with reduced cross-bridge cycling rates. A higher cell Ca2+ concentration appears to be necessary to activate myosin kinase and produce myosin phosphorylation than is needed for force maintenance. We suggest that agonist-induced Ca2+ transients, coupled with the differential Ca2+ sensitivity of the two regulatory systems, may explain the observed transient in myosin phosphorylation during a maintained contraction in smooth muscle.


Mechanical distraction for the treatment of posttraumatic stiffness of the elbow in children and adolescents. Surgical technique.
Elbow contracture is a recognized sequela of elbow injuries in children and adolescents, but previous studies of operative treatment with formal capsular release have demonstrated unpredictable outcomes and unfavorable results. Over a period of five years, fourteen children and adolescents with a mean age of fourteen years who had posttraumatic stiffness of the elbow were managed according to a prospective protocol. Eleven patients had undergone a mean of three previous operative procedures before the index operation. After intraoperative distraction with an external fixator, there was a relaxation phase for six days followed by mobilization of the elbow joint under distraction in the fixator for a mean of seven weeks. Intraoperative range of motion under distraction reached a mean of 100 degrees . Open arthrolysis was not performed, but in four children impinging heterotopic bone was removed through a limited approach. Decompression of the ulnar nerve was performed in seven patients. The mean preoperative arc of total elbow motion was 37 degrees . The mean pronation was 46 degrees , and the mean supination was 56 degrees . After a mean duration of follow-up of thirty-four months, all patients but two had achieved an arc of motion of 100 degrees . The mean arc of flexion-extension was 108 degrees (range, 75 degrees to 130 degrees ). The mean range of pronation was 73 degrees (range, 20 degrees to 90 degrees ), and the mean range of supination was 75 degrees (range, 10 degrees to 90 degrees ). There were no pin-track infections or deep infections, and all elbows were stable. At the time of follow-up, three patients had radiographic evidence of humeroulnar degeneration. Closed distraction of the elbow joint with use of a monolateral external fixation frame with motion capacity yields more favorable results than other previously reported options for the treatment of posttraumatic elbow contractures in children and adolescents.


Experimental and Computational Models for Simulating Sound Propagation Within the Lungs.
An acoustic boundary element model is used to simulate sound propagation in the lung parenchyma and surrounding chest wall. It is validated theoretically and numerically and then compared with experimental studies on lung-chest phantom models that simulate the lung pathology of pneumothorax. Studies quantify the effect of the simulated lung pathology on the resulting acoustic field measured at the phantom chest surface. This work is relevant to the development of advanced auscultatory techniques for lung, vascular, and cardiac sounds within the torso that utilize multiple noninvasive sensors to create acoustic images of the sound generation and transmission to identify certain pathologies.


A Huge Ovarian Dermoid Cyst: Successful Laparoscopic Total Excision.
Giant ovarian cysts, ≥15 cm in diameter, are quite rare in women of reproductive age. Here, we present a case of ovarian cyst with unusual presentation treated by laparoscopic surgery. On histology, mass was found to be mature cystic teratoma. The diagnostic and management challenges posed by this huge ovarian cyst were discussed in the light of the literature.


[New method for measurement of size and concentration of lyposomes].
To develop new method of determination of size and concentration of lyposomes based on measurement of opacity in dispersed media. Dispersions of lyposomes from dipalmitoylposphatidylcholine were the object of the study. Opacity spectrum of lyposome suspension was measured with Zenyth 200st spectrophotometer. Mean values of diameter of lyposomes determined by opacity spectrum were compared with the same values measured by electron microscopy (JEM 100-CX, JEOL, Japan) with magnification 58,000 - 100,000. Refraction index was measured with refractometer RPL-3 (Russia). Sizes of lyposomes measured by the new method and by electron microscopy did not differ significantly. Determination of sizes and concentration of lyposomes by the new method did not depend from effect of secondary multiple scattering of light. Obtained data allowed to conclude that developed method could be used in practice. Advantages of the new method are usage of common spectrophotometers and photoelectrocolorimeters for deriving of liposomes suspension opacity curve as well as its high validity, which are confirmed by data obtained with electron microscopy.


Evaluation of Seasonal Changes in Facial Skin With and Without Acne.
The objective of this study was to compare facial skin of adolescent males with (acne) and without acne (non-acne) over the course of 1 year. At study entry, presence of acne was determined by clinical image analysis (acne n=7, non-acne n=10). Monthly evaluations of skin condition were made using standard and fluorescent imaging, fluorescence spectroscopic analysis, sebum analysis, skin high frequency conductivity (moisture content), transepidermal water loss (TEWL), and sampling of skin bacteria (aerobic and anaerobic). Data were evaluated seasonally. Over the course of the study, subjects in the acne and non-acne groups had no significant increase in their clinical acne score. Sebum production was significantly greater in subjects with acne than in those without for each season examined (P<0.019) and was lowest in the winter and highest in the fall. TEWL was higher in those with acne than without acne across all seasons (P=0.001). Skin moisture in both groups was increased during summer and fall compared with winter (P≤0.016 for both seasons). Subjects with acne had a higher recovery of both aerobic and anaerobic bacteria compared with subjects without acne (P≤0.015). Analysis of cheek skin in the nasal area revealed significantly higher fluorescence (500-800 nm) in image-based and spectroscopic analysis from subjects with acne, suggesting the greater presence of the bacterial metabolite porphyrin in those with acne. In these cohorts of adolescent males, significant differences in sebum production, skin barrier function, moisture content, and microbial load (anaerobic and aerobic) were noted between those with and without acne. Evidence for seasonality was observed, with lower lipid production and reduced barrier function during the winter. More studies to quantify differences in skin lipid components and bacterial species among these cohorts are planned.


Recurrence in major depressive disorder: a neurocognitive perspective.
Depressive disorders are amongst the leading causes of disability and mortality worldwide and, as such, it is predicted that by 2010 only cardio-ischaemic disorders will provide a greater burden. In addition to the sizable emotional, individual and social burden, depressive disorders cost an estimated US$83.1 billion per year in the United States alone. In spite of effective treatments, a large proportion of sufferers go on to experience recurrences. With successive recurrences, the likelihood of subsequent episodes increases. Despite this, research to date has tended to focus on first episodes or else has not distinguished between episodes. This editorial review highlights a number of differences between first and recurrent episodes which, in turn, recommend more longitudinal, recurrence-oriented, treatments. We also examine the findings from acute tryptophan depletion studies which, it is speculated, help to understand the differences between successive episodes. The overall aim, however, is to highlight the importance of recurrence in depression and to stimulate debate.


Clinical and hematologic features of 300 patients affected by hereditary spherocytosis grouped according to the type of the membrane protein defect.
Hereditary spherocytosis is a very heterogeneous form of hemolytic anemia. The aim of this study was to relate the type of molecular defect with clinical and hematologic features and response to splenectomy using information from a large database of patients. Data from 300 consecutive patients with hereditary spherocytosis, grouped according to the results of sodium dodecyl sulphate-polyacrylamide gel electrophoresis, were analyzed and the sensitivity of red cell osmotic fragility tests was compared in various subsets of patients. Band 3 and spectrin deficiencies were the most common protein abnormalities (54% and 31%, respectively); 11% of cases were not classified by the electrophoretic analysis. Spectrin deficiency was more frequently diagnosed in childhood and band 3 deficiency in adulthood. Hemoglobin concentration was slightly lower, spherocyte number and hemolysis markers higher in spectrin deficiency than in band 3 deficiency. The sensitivity of the osmotic fragility tests ranged from 48% to 95%, and was independent of the type and amount of the membrane defect. The association of the acidified glycerol lysis test and the NaCl test on incubated blood reached a sensitivity of 99%. Splenectomy corrected the anemia in patients with all subtypes of hereditary spherocytosis although spectrin-deficient patients still showed increased reticulocyte numbers and levels of unconjugated bilirubin. Splenectomy allowed the identification of the membrane defect in all the previously unclassified patients, most of whom had spectrin and/or ankyrin deficiency. The definition of the red cell membrane defect in hereditary spherocytosis has no major clinical implications, but may be useful for a differential diagnosis from other hematologic disorders that mimic this hemolytic anemia.


Hypertension genetic risk score is associated with burden of coronary heart disease among patients referred for coronary angiography.
Recent GWAS studies have identified more than 300 SNPs associated with variation in blood pressure. We investigated whether a genetic risk score constructed from these variants is associated with burden of coronary heart disease. From 2010-2014, 4,809 individuals admitted to coronary angiography in Capital Region of Copenhagen were genotyped. We calculated hypertension GRS comprised of GWAS identified SNPs associated with blood pressure. We performed logistic regression analyses to estimate the risk of hypertension and prevalent CHD. We also assessed the severity of CHD associated with the GRS. The analyses were performed using GRS quartiles. We used the Inter99 cohort to validate our results and to investigate for possible pleiotropy for the GRS with other CHD risk factors. In COGEN, adjusted odds ratios comparing the 2nd, 3rd and 4th cumulative GRS quartiles with the reference were 1.12(95% CI 0.95-1.33), 1.35(95% CI 1.14-1.59) and 1.29(95% CI 1.09-1.53) respectively, for prevalent CHD. The adjusted multinomial logistic regression showed that 3rd and 4th GRS quartiles were associated with increased odds of developing two(OR 1.33, 95% CI 1.04-1.71 and OR 1.36, 95% CI 1.06-1.75, respectively) and three coronary vessel disease(OR 1.77, 95% CI 1.36-2.30 and OR 1.65, 95% CI 1.26-2.15, respectively). Similar results for incident CHD were observed in the Inter99 cohort. The hypertension GRS did not associate with type 2 diabetes, smoking, BMI or hyperlipidemia. Hypertension GRS quartiles were associated with an increased risk of hypertension, prevalent CHD, and burden of coronary vessel disease in a dose-response pattern. We showed no evidence for pleiotropy with other risk factors for CHD.


Habituation of the threatening response in cats and kittens.
Modifications of the threatening response occurring in cats and naive kittens were studied during repeated confrontations by a dog. Most of the kittens when confronted by a dog for the first time displayed a threatening response. There was a wide individual variation in the intensity of the first response. In some kittens initial sensitization occurred. With repetition of the confrontations the response waned. Complete habituation of the response in the kittens was achieved after 4 to 15 daily sessions, in adult cats it took from 2 to 47 sessions. In both kittens and adult cats various components, of the threatening response habituated at a different rate. This caused the pattern of the response to change during repetitive confrontations. This out of step habituation of the components has been described with regard to the cardiovascular components of the response (8). It is concluded that the threatening reaction is subject to habituation and that the components of this response may be suppressed independently from the others, thus modifying the pattern of the response.


Effects of West Nile virus dose and extrinsic incubation temperature on temporal progression of vector competence in Culex pipiens quinquefasciatus.
Culex pipiens quinquefasciatus were fed blood containing either 7.0 +/- 0.1 logs plaque-forming units (pfu)/ml (high dose) or 5.9 +/- 0.1 logs pfu/ml (low dose) of West Nile virus and held at extrinsic incubation temperatures (EIT) of 28 degrees C or 25 degrees C. Approximately 20 mosquitoes per dose were collected after incubation periods (IP) of 4, 6, 8, and 12 days postinfection (dpi). Infection rates were influenced by EIT and virus dose but not by IP. Body titer was significantly higher for mosquitoes fed the high dose and held at 28 degrees C at the later IPs (6, 8, and 12 dpi). However, leg titer was significantly higher for mosquitoes at the later IPs but did not differ between EITs or doses. Because infection rates varied with EIT and dose, there is likely a midgut infection barrier influenced by these factors that is not influenced by IP. Dissemination rates were influenced by all 3 factors consistent with the presence of a midgut escape barrier. Dissemination rate, body titer, and leg titer were dependent on IP, indicating the need to investigate multiple time points in vector competence studies to elucidate critical events in infection and dissemination.


[ADH and cAMP receptor binding in the kidney medulla of mice insensitive to ADH].
Specific binding of ADH by the membrane fraction of the kidney medulla was lower in the normal CBA mice than in mutant mice with nephrogenic diabetes. Gel filtration of the solubilized ADH receptors of mutants revealed the presence of an unidentified factor which caused cooperative binding of ADH. DEAE-chromatography revealed no difference between cytosolic cAMP receptors in normal and mutant animals. Assay of GTP-ase activity of the membrane fraction revealed that ADH increased this parameter in CBA mice but not in mutant animals. Cholera toxin significantly diminished membrane ATP-ase activity whereas membrane preparations from mutant mice developed a reactivity to ADH. GTP binding ability in these preparations was higher than inn intact ones. In CBA mice this ability increased dramatically. HPLC profiles of G-protein complexes with GNP were very different in CBA and mutant mice. Mutation seems to cause changes both in binding and in "cross-talk" link op-complex membrane receptor of ADH.


Influence of vitamin D on sarcopenia pathophysiology: A longitudinal study in humans and basic research in knockout mice.
Vitamin D is an essential nutrient in musculoskeletal function; however, its relationship to sarcopenia remains ambiguous, and the mechanisms and targets of vitamin D activity have not been elucidated. This study aimed to clarify the role of vitamin D in mature skeletal muscle and its relationship with sarcopenia. This epidemiological study included 1653 community residents who participated in both the fifth and seventh waves of the National Institute for Longevity Sciences, Longitudinal Study of Aging and had complete background data. Participants were classified into two groups: vitamin D-deficient (serum 25-hydroxyvitamin D < 20 ng/mL) and non-deficient (serum 25-hydroxyvitamin D ≥ 20 ng/mL); they underwent propensity-score matching for background factors (age, sex, height, weight, comorbidities, smoker, alcohol intake, energy intake, vitamin D intake, steps, activity, season and sarcopenia). Changes in muscle strength and mass over the 4-year period were compared. For basic analysis, we generated Myf6CreERT2 Vitamin D Receptor (VDR)-floxed (VdrmcKO ) mice with mature muscle fibre-specific vitamin D receptor knockout, injected tamoxifen into 8-week-old mice and analysed various phenotypes at 16 weeks of age. Grip strength reduction was significantly greater in the deficient group (-1.55 ± 2.47 kg) than in the non-deficient group (-1.13 ± 2.47 kg; P = 0.019). Appendicular skeletal muscle mass reduction did not differ significantly between deficient (-0.05 ± 0.79 kg) and non-deficient (-0.01 ± 0.74 kg) groups (P = 0.423). The incidence of new cases of sarcopenia was significantly higher in the deficient group (15 vs. 5 cases; P = 0.039). Skeletal muscle phenotyping of VdrmcKO mice showed no significant differences in muscle weight, myofibre percentage or myofibre cross-sectional area; however, both forelimb and four-limb muscle strength were significantly lower in VdrmcKO mice (males: forelimb, P = 0.048; four-limb, P = 0.029; females: forelimb, P < 0.001; four-limb, P < 0.001). Expression profiling revealed a significant decrease in expression of sarcoendoplasmic reticulum Ca2+ -ATPase (SERCA) 1 (P = 0.019) and SERCA2a (P = 0.049) genes in the VdrmcKO mice. In contrast, expression of non-muscle SERCA2b and myoregulin genes showed no changes. Vitamin D deficiency affects muscle strength and may contribute to the onset of sarcopenia. Vitamin D-VDR signalling has minimal influence on the regulation of muscle mass in mature myofibres but has a significant influence on muscle strength.


The origin of the Tibetan Mastiff and species identification of Canis based on mitochondrial cytochrome c oxidase subunit I (COI) gene and COI barcoding.
DNA barcoding is an effective technique to identify species and analyze phylogenesis and evolution. However, research on and application of DNA barcoding in Canis have not been carried out. In this study, we analyzed two species of Canis, Canis lupus (n = 115) and Canis latrans (n = 4), using the cytochrome c oxidase subunit I (COI) gene (1545 bp) and COI barcoding (648 bp DNA sequence of the COI gene). The results showed that the COI gene, as the moderate variant sequence, applied to the analysis of the phylogenesis of Canis members, and COI barcoding applied to species identification of Canis members. Phylogenetic trees and networks showed that domestic dogs had four maternal origins (A to D) and that the Tibetan Mastiff originated from Clade A; this result supports the theory of an East Asian origin of domestic dogs. Clustering analysis and networking revealed the presence of a closer relative between the Tibetan Mastiff and the Old English sheepdog, Newfoundland, Rottweiler and Saint Bernard, which confirms that many well-known large breed dogs in the world, such as the Old English sheepdog, may have the same blood lineage as that of the Tibetan Mastiff.


Algorithm for advanced canonical coding of planar chemical structures that considers stereochemical and symmetric information.
We describe a rigorous and fast algorithm for advanced canonical coding of planar chemical structures based on the algorithm of Faulon et al. (J. Chem. Inf. Comput. Sci. 2004, 44, 427-436). Our algorithm works well even for highly symmetric structures; moreover, an advantage of our algorithm includes providing a rigorous canonical numbering of atoms with a consideration of stereochemistry and recognizing symmetric moieties. The planar structural line notation with the canonical numbering is also fit for use with stereochemical line notation. These capabilities are usable for general purposes in chemical structural coding and are particularly essential for detecting equivalent atoms in NMR studies. This algorithm was implemented on a 13C NMR chemical shift prediction system CAST/CNMR. Applications of the algorithm to several organic compounds demonstrate the practical efficiency of the rigorous coding.


Who is providing HIV testing services? The profile of lay counsellors providing HIV testing services in Johannesburg, South Africa in the treat-all era.
Lay counsellors are critical in sustaining access to HIV testing services (HTS) and psychosocial support for persons living with HIV (PLHIV). We aimed to describe the professional and psychosocial profiles of lay counsellors in primary healthcare (PHC) clinics in Johannesburg, South Africa under the universal-test-and-treat (UTT) policy context. We conducted a descriptive analysis of a cross-sectional survey among adult (≥ 18 years) lay counsellors from 20 PHC facilities (2-3/ clinic) in Johannesburg, South Africa. Consenting counsellors were interviewed between June 2018 and March 2019. We report on counsellors' demographic profiles, training, work experience, and mental and emotional well-being. Overall, 55 consenting adult (≥ 18 years) lay counsellors (92.7% female, median age 37 years, interquartile range [IQR]: 33-44, and 27.3% HIV diagnosed) were surveyed. Most (85.5%) were Department of Health lay counsellors receiving a volunteer stipend at the time. Overall, 56.4% had been working as counsellors for five years or longer. The majority (87%) had completed the National HIV Testing Services Policy Guidelines-recommended 10-day basic counselling training, but 45.2% had not completed refresher training within the guideline's required 24 months. Reported operational barriers include lack of designated space for counselling (56.4%), inadequate professional supervision and support (40.7%) and insufficient emotional support (over 56.4%), and 60% were overwhelmed by their workload. A total of 18.2% had major depressive symptoms, and the same proportion scored low for psychological well-being. While most (87.3%) reported moderate job satisfaction, 50.9% actively sought alternative employment. Despite lay counsellors' significant role in delivering HIV care in South Africa, there has been minimal investment in their skills development, emotional support, and integration into the formal health workforce. Counsellors' persisting unmet psychosocial, training, and professional needs could impact their efficacy in the UTT era.


The Vanishing Twin Syndrome: Two Cases of Extreme Malformations Associated With Vanished Twins.
Two cases of devastating fetal malformations associated with vanished monochorionic twins were identified upon review of pathology files. A 35-year-old G1P0 woman and 36-year-old G3P1 woman were both diagnosed with an intrauterine twin gestation via transvaginal ultrasound at 10 weeks. The spectrum of fetal anomalies ranged from omphalocele, bilateral upper extremity, and unilateral lower extremity hypoplasia, to craniofacial malformation with diaphragmatic hernia. On histopathologic examination, the placentas demonstrated vascular anastomoses between the surviving co-twin and the "vanished" fetal sac. We propose anastomotic placental vasculature as a contributing factor to the observed fetal malformations. Additionally, genetic or teratogenic factors may have been attributed to the demise of the first twin and the anomalies seen in the other twin. While such instances are rare, they are important to consider when counseling patients regarding outcomes associated with a monochorionic vanished twin.


The social construction of identity in HIV/AIDS home-based care volunteers in rural KwaZulu-Natal, South Africa.
Home-based care volunteer (HBCV) identity and how it is shaped was the main focus of the study. Fifteen HBCVs were interviewed about their work and personal life stories and then interviewed reflectively using a narrative interviewing style. Specific attention was paid to contextual meta-narratives and social field narratives in understanding the women's stories. Findings indicate that social field narratives of the women's stories were dominated by negative aspects of gender, poverty and socio-political factors. These were seen to coincide with the 'feminisation of responsibility' in this context effectively coercing the women into agency which manifested as their home-based care work. Meta-narratives influencing the women's lives were dominated by stories of communal motherhood, aspirations to service-oriented work and religious beliefs and commitment. The question of how it is possible for women who are seemingly constrained by oppressive narratives to voluntarily engage in community participation was answered in the women's personal life stories about being compassionate, hopeful, helpful and ambitious and having initiative. These characteristics collectively pointed to personal agency. Exploring connections between the different aspects of identity and context revealed that the women made sense of their community participation through their personal identities as strong and loving mothers. Connections between volunteer personal identity, agency and volunteer group identity were explored to make sense of the link between HBCV identity and volunteerism. The mother identity, encompassing personal agency (strength or power) and love (the meta-narrative of communal motherly love), was salient in influencing community participation of the group.


[Sigmund Freud and Hans Blüher in up to now unpublished letters].
In the years 1912 and 1913 there occured an epistolary encounter between the founder-father of psychoanalysis and the young Hans Bluher who had been active in the Wandervogel movement, a German youth movement with strong traditionalist and nationalist leanings. Their correspondence centered around the evaluation of male homosexuality, a point on which Freud and Bluher were not in entire agreement. In his introduction Neubauer outlines the intellectual nub of this debate and sketches Bluher's later career and his gradual transformation into a biologistically motivated anti-Semite completely and utterly disowned by Freud.


Filamentous inclusions of unusual composition and architecture in a metastatic tumor showing myoepithelial differentiation.
A hyaline/eosinophilic cytoplasmic inclusion is described in a metastatic tumor occurring in the omentum and bowel of a 32-year-old white woman. The tumor was essentially of round cell type; was positive for cytokeratin, actin, and S-100 protein by immunohistochemistry; and showed desmosomes (maculae adherentes), tonofibrils, modestly developed myofilaments with focal densities, and a basal lamina by electron microscopy. It was therefore interpreted as showing myoepithelial differentiation. Nearly all cells possessed a large, rounded cytoplasmic inclusion that was architecturally unusual, consisting of intermediate filaments intermingled with lattices of fine filaments. The inclusions exhibited an additionally unusual immunohistochemical staining for both cytokeratin and actin. A comparison with cases described in the literature is made.