Alpha1-adrenoceptor subtypes involved in vasoconstrictor responses to exogenous and neurally released noradrenaline in rat femoral resistance arteries.
1. The alpha(1)-adrenoceptor subtypes involved in responses to exogenous and neurally released noradrenaline in rat femoral resistance arteries were characterised using a small vessel myograph, with antagonists prazosin (nonsubtype selective), 5-methyl-urapidil (alpha(1A)-selective), BMY 7378 (alpha(1D)-selective) and the alkylating agent chloroethylclonidine (preferential for alpha(1B)-). 2. Prazosin and 5-methyl-urapidil produced rightward shifts of the exogenous noradrenaline concentration - response curve (CRC) with pA(2) values of 9.2 and 9.1 respectively, in agreement with the presence of alpha(1A)-adrenoceptors. BMY 7378 (1 microm) shifted the noradrenaline CRC with an apparent pK(B) of 6.7, in agreement with the presence of alpha(1A)-, but not alpha(1D)-, adrenoceptors. Chloroethylclonidine at 1 microm had no effect and at 10 microm produced only a small reduction (c. 20%) in the maximum response to noradrenaline, indicating little, if any, contribution from alpha(1B)-adrenoceptors. 3. Responses of the rat femoral resistance arteries to electrical field stimulation (EFS) at 5-30 Hz for 10 s and 0.05 ms pulse width were principally due to alpha(1)-adrenoceptor stimulation. Prazosin and 5-methyl-urapidil inhibited EFS-mediated responses with pIC(50)s of 9.3 and 8.2, respectively, consistent with the alpha(1A)-adrenoceptor being the predominant subtype. Responses to EFS at 10-30 Hz were relatively insensitive to BMY 7378 (pIC(50), 6.5-6.7), while responses to 5 Hz were inhibited with a significantly higher pIC(50) of 8.02, suggesting the contribution of alpha(1D)-adrenoceptors. Chloroethylclonidine had no effect on responses to EFS, ruling out the contribution of an alpha(1B)-subtype. In the presence of cocaine, the predominant subtype involved in responses to EFS was the alpha(1A)-adrenoceptor, with a contribution from alpha(1D)-adrenoceptors at low frequency, as seen in the absence of cocaine. However, there was also a significant increase in the sensitivity to BMY 7378 at higher frequencies, suggesting that a further small alpha(1D)-adrenoceptor component may be uncovered in the presence of cocaine. 5. The present study has shown a predominant role of the alpha(1A)-adrenoceptor in contractions due to exogenous noradrenaline and to neurally released noradrenaline in rat femoral resistance arteries. alpha(1D)-Adrenoceptors are not involved in responses to exogenous noradrenaline but appear to be activated by neurally released noradrenaline at a low frequency of stimulation and at higher frequencies in the presence of neuronal-uptake blockade.


Expanded bodipy dyes: anion sensing using a bodipy analog with an additional difluoroboron bridge.
Oxalyl-tethered pyrroles can be doubly bridged with two difluoroboron chelating units to yield bright orange dyes. Interestingly, in polar organic solvents, the addition of fluoride and cyanide result in reversible detachment of the otherwise stable difluoroboron bridges, resulting in sharp changes in color. Thus, this novel compound behaves as a highly selective chromogenic sensor for fluoride and cyanide ions.


Analyzing the far-right political action on Twitter: the Chilean constituent process.
The concept of "politics of the end" assumes the catastrophe of living in a world that produces new forms of accumulation and allows symbolic and semiotic capital to create value. Currently, various far-right movements worldwide seem to appropriate this concept, employing radical communication strategies as a repertoire to contest the public agenda. These strategies include the massive creation of bots on social networks to spread hate speech and coordinate ideological manifestations. This article seeks to verify the use of these strategies by the Chilean far-right on Twitter. For the above, a social network analysis approach is proposed during the current socio-political crisis in Chile, which began with the massive protests of October 2019 and led to an unprecedented constituent process. For nine months, we studied five opinion leaders on Twitter from the Chilean far-right, who together have more than 600 thousand followers and almost 130 thousand followings. Through descriptive, quantitative, and qualitative techniques, an explicit political action "from the resistance" is revealed in the activity of the network, which includes hundreds of new users and coordinated bots to disseminate identifiable discourses with strongly ideological ideas. This coordination also presents identifiable differences in how opinion leaders interact and communicate with their network environment.


Long-Term Results from the Pivotal Multicenter Trial of Ultrasound-Guided Percutaneous Arteriovenous Fistula Creation for Hemodialysis Access.
To report the 5-year results from the Pivotal Multicenter Trial of Ultrasound-Guided Percutaneous Arteriovenous Fistula (pAVF) Creation for Hemodialysis Access. The retrospective review of 107 intent-to-treat (ITT) patients from the pivotal trial provided a long-term follow-up population (LTP) of 85 patients with a median follow-up duration of 50 months (range, 12-60 months). Data evaluated in the LTP group were fistula maturation and usage, secondary procedures, and complications. The Kaplan-Meier analysis of primary patency, assisted primary patency, cumulative patency, and functional patency (time from 2-needle cannulation to abandonment) were performed for the ITT population. In the LTP, 99% (84 of 85) of fistulae were mature, with 99% (78 of 79) of patients requiring hemodialysis using their pAVF. Sustained fistula use (2-needle cannulation at the prescribed rate, 2 of 3 sessions) was achieved in 92% (78 of 85) of patients, with 7 patients not using their pAVF because they were not on dialysis (n = 4), were on peritoneal dialysis (n = 2), and refused to use fistula (n = 1). Fistula maintenance was required in 31.8% (27 of 85) of patients and included fistula dysfunction (21.2%), thrombosis (5.9%), cannulation injury (12.9%), and arm swelling (4.7%). The number of procedures performed per patient per year to maintain function and patency was 0.32 (91 of 288) for years 2-5. The cumulative patency rates were 89.5%, 88.4%, 88.4%, 85.6%, and 82.0% for years 1, 2, 3, 4, and 5, respectively. The functional patency was 91.8% at the end of the study. There were no major complications related to pAVF during the long-term follow-up. Percutaneous fistulae have provided clinically effective and durable access for hemodialysis with low complications. The continued use and evaluation of pAVF are warranted.


Isolation and characterization of a growth factor (embryonin) from bovine fetuin which resembles alpha 2-macroglobulin.
A serum-free hormone-supplemented medium which was previously formulated for the growth of mouse embryonal carcinoma (EC) cells and rat mammary epithelial (REM) cells required the presence of crude bovine fetuin as a medium supplement for maintaining cell growth. This requirement could not be replaced by purified fetuin preparations. The major growth-promoting activity (embryonin) in the crude fetuin preparation has been purified to near homogeneity by carboxymethylcellulose chromatography and high performance gel filtration chromatography. Purified embryonin, 2 to 4 micrograms/ml, is able to stimulate the growth of mouse EC cells in a serum-free hormone-supplemented medium to a level that is achieved with 0.5 to 1 mg/ml of the crude fetuin preparation. The biological activity resides in a high molecular weight glycoprotein (Mr = 270,000). Three polypeptide chains are observed following reduction, a major polypeptide (Mr = 185,000) and two minor chains (Mr = 116,000 to 120,000 and 68,000). Embryonin differs from pure fetuin in molecular weight, isoelectric point, amino acid composition, and immunological reactivity. However, embryonin is similar to human alpha 2-macroglobulin (alpha 2M) in these physicochemical and immunological properties. Resemblance to alpha 2M is also indicated by the observation that alpha 2M used over the same protein concentration range as embryonin produces a comparable stimulation of EC cell growth in the absence of serum. In addition, embryonin and alpha 2M produce a 2- to 10-fold differential stimulation of collagen production in cultures of normal rat kidney and RME cells, a response which may be linked to the growth-promoting activity of these two proteins.


Density functional study of S(N) 2 substitution reactions for CH(3) Cl + CX(1) X(2•-) (X(1) X(2) = HH, HF, HCl, HBr, HI, FF, ClCl, BrBr, and II).
A systematic investigation on the S(N) 2 displacement reactions of nine carbene radical anions toward the substrate CH(3) Cl has been theoretically carried out using the popular density functional theory functional BHandHLYP level with different basis sets 6-31+G (d, p)/relativistic effective core potential (RECP), 6-311++G (d, p)/RECP, and aug-cc-pVTZ/RECP. The studied models are CX(1) X(2•-) + CH(3) Cl → X(2) X(1) CH(3) C(•) + Cl(-) , with CX(1) X(2•-) = CH(2) (•-) , CHF(•-) , CHCl(•-) , CHBr(•-) , CHI(•-) , CF(2) (•-) , CCl(2) (•-) , CBr(2) (•-) , and CI(2) (•-) . The main results are proposed as follows: (a) Based on natural bond orbital (NBO), proton affinity (PA), and ionization energy (IE) analysis, reactant CH(2) (•-) should be a strongest base among the anion-containing species (CX(1) X(2•-) ) and so more favorable nucleophile. (b) Regardless of frontside attacking pathway or backside one, the S(N) 2 reaction starts at an identical precomplex whose formation with no barrier. (c) The back-S(N) 2 pathway is much more preferred than the front-S(N) 2 one in terms of the energy gaps [ΔE cent≠(front)-ΔE cent≠(back)], steric demand, NBO population analysis. Thus, the back-S(N) 2 reaction was discussed in detail. On the one hand, based on the energy barriers (ΔE cent≠ and ΔE ovr≠) analysis, we have strongly affirmed that the stabilization of back attacking transition states (b-TSs) presents increase in the order: b-TS-CI(2) < b-TS-CBr(2) < b-TS-CCl(2) < b-TS-CHI < b-TS-CHBr < b-TS-CHCl < b-TS-CF(2) < b-TS-CHF < b-TS-CH(2) . On the other hand, depended on discussions of the correlations of ΔE ovr≠ with influence factors (PA, IE, bond order, and ΔE def≠), we have explored how and to what extent they affect the reactions. Moreover, we have predicted that the less size of substitution (α-atom) required for the gas-phase reaction with α-nucleophile is related to the α-effect and estimated that the reaction with the stronger PA nucleophile, holding the lighter substituted atom, corresponds to the greater exothermicity given out from reactants to products.


Carotenoid Isotopolog Profiling in 13C-Labeled Leaf Extracts by LC-MS and LC-FTICR-MS.
Mass spectrometry (MS)-based metabolite analysis combined with stable isotope labeling offers a powerful tool to study dynamic regulation of metabolic pathways and metabolite fluxes in biological systems. Here we describe a method to analyze the composition of carotenoid isotopologs in 13C-labeled leaf extracts by using liquid chromatography (LC)-MS and LC-Fourier transform ion cyclotron resonance (FTICR)-MS. High mass resolution of the latter enables unambiguous assignment of observed mass to a unique chemical formula. Based on peak intensity the relative abundance and the degree of 13C labeling are calculated for individual carotenoid isotopologs.


Management of Cutaneous Immune-Related Adverse Events in Patients With Cancer Treated With Immune Checkpoint Inhibitors: A Systematic Review.
There exists a paucity of literature that summarizes the effective management of cutaneous immune-related adverse events (cirAEs) in patients with cancer who are receiving immune checkpoint inhibitors (ICIs). Most published articles are small case series from a single institution. To our knowledge, the spectrum of possible treatments has not been systematically reviewed to highlight the breadth of options when caring for patients with cirAEs. To further characterize the development of subtypes of cirAEs in patients with cancer treated with ICIs and provide recommendations on optimal treatment regimens based on the current literature. A search was performed in PubMed, Embase European, Web of Science, and Google Scholar on June 26, 2020, according to Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) reporting guidelines, limited to the years 2010 to 2020. Articles that met predetermined inclusion criteria (published between January 1, 2010, and June 1, 2020; written in the English language; and original articles, brief reports, case reports, and research letters that reported primarily on cirAE management) were selected, and data were abstracted. Articles that met the scope of the review were also added from reference lists. When possible, the results of studies that addressed a similar question were combined quantitatively. In total, 138 studies (87 from the aforementioned literature search and 51 additional studies pulled from the reference lists of included articles) were included that reported on 879 cirAEs. The subtypes of cirAEs included maculopapular, pruritus, lichenoid, immunobullous, psoriasiform, granulomatous, erythema multiforme or Stevens Johnson Syndrome, drug rash with eosinophilia and systemic symptoms, connective tissue disease, hair, oral, and miscellaneous. Treatments for cirAEs included a combination of topical corticosteroids, systemic corticosteroids, steroid-sparing agents, and discontinuation or cessation of immunotherapy. This systematic review found that treatment with ICIs was associated with many types of skin toxic effects, each with unique treatment options beyond current published guidelines. Further research into key differences between subtypes is critical to improve the care provided to patients with cancer.


The influence of brain cytosol on RNA synthesis and RNA products of isolated mouse brain nuclei.
The incubation of isolated nuclei obtained from 10-day-old mouse brain in the presence of brain cell cytosol resulted in an increase in the synthesis of RNA. Under conditions of saturating concentrations of nucleoside triphosphates, the influence of cytosol could not be duplicated by the addition of cyclic nucleotides. The stimulatory activity of cytosol on brain nuclear RNA synthesis could not be attributed to either alterations in the permeability of the nuclear envelope or an increased uptake of radioactively-labeled precursors. Sucrose gradient analysis demonstrated that the RNA products synthesized by nuclei isolated from 10-day-old and adult mouse brain were of a relatively low molecular weight. However, the addition of cytosol resulted in a significant increase in the size of the RNA transcripts. In contrast to the observations with 10-day-old and adult brain nuclei, the RNA from 2-day-old mouse brain nuclei was larger in size and relatively unaffected by the presence of cytosol. Although cytosol caused an increase in the amounts of poly[A]-RNA in nuclei of 2-day-old and adult animals, no comparable effect could be measured in nuclei from 10-day-old brain tissue.


Ultrastructural alterations in human blood leukocytes induced by porcine circovirus type 1 infection.
Swine infectious pathogens, especially viruses, represent a potential public health risk associated with the use of pig tissues for xenotransplantation in humans. We hypothesized that porcine circovirus type I (PCV-1) may infect human mononuclear cells, resulting in ultrastructural alterations of the target cells. Transmission electron microscopy was used for evaluating ultrastructural alterations of human cells exposed to a PCV-infected PK15 cell line. A polymerase chain reaction (PCR) assay and fluorescence in situ hybridization (FISH) were developed for detecting PCV-1 in human mononuclear cells. Morphological alterations of the human T cells exposed to PCV PK15 showed ''boomerang-shaped'' intracytoplasmic inclusions. Nucleocapsids appeared free, close to the nucleus, or contained into cytoplasmic vacuoles. Virions were observed near the surface of the human cells. A considerable number of mature virions and immature forms could be observed in the human cells that had a completely intact nuclear membrane with no alteration in the disposition of chromatin. PCV-1 particles were identified budding into typical Golgi saccules and vacuoles. Virions sized up to 23 nm in diameter, and appeared in the nucleus and in the periphery of the cellular core. PCV-1 infection was detected on CD4+, CD8+, CD14+, CD19+, and CD56+ human cells by PCR assay and FISH. These results suggest that PCV has the capability of infecting human leukocytes in vitro, and should be considered a potential risk of viral transmission during xenotransplantation.


Relationship between vertical ground reaction force and muscle strength while climbing stairs after total hip arthroplasty.
Little is known about the relationship between vertical ground reaction force (vGRF) and muscle strength while climbing stairs after total hip arthroplasty (THA). We investigated the relationship between muscle strength and vGRF during stair climbing in 21 women with THA. Differences in the time required to climb stairs and side-to-side differences in muscle strength and vGRF during climbing were examined at 6 and 12 months. The relationship between climbing time and vGRF variables and that between vGRF and muscle strength were analyzed using Spearman's rank correlation coefficient at 6 and 12 months. First peak force (Fz1) was defined as the peak vGRF at heel strike normalized by body weight. Climbing time was related to the time to Fz1 at 6 and 12 months post-THA (r = 0.50-0.64). At 6 months, time to Fz1 was related to the strength of muscles involved in hip extension in the operated limb (r = -0.46). At 12 months, time to Fz1 was related to the strength of muscles responsible for knee extension in the operated limb (r = -0.45). At 6 months, climbing stairs depended on muscle strength in the hip extensors. Conversely, at 12 months, the functional recovery of knee extensors was important for transferring weight upward while climbing stairs. Therefore, to improve the climbing function of patients with THA, it is important to not only focus on the recovery of muscle strength in the hip extensors but also improve muscle strength in the knee extensors.


Biomarkers of Type IV Collagen Turnover Reflect Disease Activity in Patients with Early-Stage Non-Alcoholic Fatty Liver (NAFL).
Identification of progressive liver disease necessitates the finding of novel non-invasive methods to identify and monitor patients in need of early intervention. Investigating patients with early-liver injury may help identify unique biomarkers. Early-liver injury is characterized by remodeling of the hepatocyte basement membrane (BM) of the extracellular matrix. Thus, we quantified biomarkers targeting two distinct neo-epitopes of the major BM collagen, type IV collagen (PRO-C4 and C4M), in patients spanning the non-alcoholic fatty liver disease (NAFLD) spectrum. We evaluated PRO-C4 and C4M in a cross-sectional study with 97 patients with NAFLD confirmed on histology. Serological levels of PRO-C4 and C4M were quantified using validated competitive enzyme-linked immunosorbent assays (ELISA). Using the fatty liver inhibition of progression (FLIP) algorithm, we stratified patients into two groups: non-alcoholic fatty liver (NAFL) and non-alcoholic steatohepatitis (NASH). Biomarker levels were investigated in the two groups in patients stratified by the NAFLD activity score (NAS). In both groups, biomarker measurements were analyzed in relation to histological scorings of steatosis, inflammation, ballooning, and fibrosis. Patients had a body mass index (BMI) of 30.9 ± 5.6 kg/m2, age of 53 ± 13 years and a NAS range of 1-8. Upon stratification by FLIP, the NASH patients had higher platelets, ALT, and AST levels than the NAFL group. Both PRO-C4 (p = 0.0125) and C4M (p = 0.003) increased with increasing NAS solely within the NAFL group; however, a large variability was present in the NASH group. Furthermore, both markers were significantly associated with lobular inflammation (p = 0.020 and p = 0.048) and steatosis (p = 0.004 and p = 0.015) in patients with NAFL. This study found that type IV collagen turnover increased with the increase in NAS in patients with NAFL; however, this was not the case in patients with NASH. These findings support the assessments of the BM turnover using biomarkers in patients with early-disease development. These biomarkers may be used to track specific processes involved in the early pathobiology of NAFL.


Validation of a method for the detection of virulent Yersinia enterocolitica and their distribution in slaughter pigs from conventional and alternative housing systems.
Various methods have been described in the literature for the detection of virulent Yersinia enterocolitica in pigs. The risk factors for pig herd contamination have yet to be determined. The objective of this study was to validate a sensitive method for the detection of Y. enterocolitica and to describe the distribution of the bacteria in pigs at slaughter from conventional and alternative ("organic") housing systems. First, samples were collected from tonsils, caecum with caecal contents, and the caecal lymph nodes of 60 slaughter pigs. These samples were used to compare the sensitivity of six different laboratory culture methods either in common use or described in the literature with that of a polymerase chain reaction with two primer pairs (multiplex PCR). Then, only PCR was used to examine tonsils, caecum and caecal lymph nodes from two groups of slaughter pigs: 210 from six conventional fattening farms and 200 from three with alternative housing. The results of the multiplex PCR were positive in 28 cases. All culture methods proved inferior to PCR in sensitivity. In the second part of the study, PCR detected 36 (18%) positive pigs from alternative housing and 60 (29%) from conventional housing (p<0.05). The highest rate of Y. enterocolitica contamination was found in tonsils (11% alternative, 22% conventional; p<0.05), followed by caecum (5%, 11%) and lymph nodes (2%, 7%). The housing system appears to be one important factor in the prevalence of this common pathogen in pig herds, as we found important differences between the two systems studied here. In the conventional system, the main risk factors appeared to be sourcing pigs from different pig suppliers, use of commercial feed and transportation to slaughter.


Erectile function after kidney transplantation: a meta-analysis.
Erectile dysfunction (ED) is common in patients with end-stage renal disease (ESRD). Whether kidney transplantation can improve erectile function in patients with ESRD is still controversial. We conducted a meta-analysis on the relationship between kidney transplantation and erectile function. A literature search was conducted on PubMed, Embase, Cochrane Library, and Web of Science until May 31, 2019. Primary outcomes were ED prevalence and each domain score of the International Index of Erectile Function (IIEF) questionnaire. We used age-matched dialysis patients or patients before kidney transplantation as a control group and compared them to kidney transplant recipients. A total of 9 articles were finally enrolled in the study. Compared with the control group, the kidney transplantation group had a lower prevalence of ED (OR 0.49, 95% CI: 0.28-0.86) and higher domain scores for erectile function (SMD 0.53, 95% CI: 0.12-0.94) and sexual desire (SMD 1.19, 95% CI: 0.11-2.27). While there were no significant variations in domain scores for orgasmic function (SMD 0.27, 95% CI: -0.10-0.63), intercourse satisfaction (SMD 0.26, 95% CI: -0.10-0.61), and overall satisfaction (SMD 0.17, 95% CI: -0.21-0.56). Patients in the kidney transplantation group had higher serum testosterone (SMD 1.20, 95% CI: 0.86-1.54) and lower prolactin (SMD -1.46, 95% CI: -2.22 to -0.69) and luteinizing hormone (SMD -0.97, 95% CI: -1.39 to -0.55). Kidney transplantation may be associated with improved erectile function in patients with ESRD. This may be attributable to the correction of endocrine hormone disorders in patients after kidney transplantation.


Transanal endoscopic microsurgical submucosal dissection (TEM-ESD) for rectal adenomas: a retrospective cohort study of 145 consecutive cases.
Transanal endoscopic microsurgery (TEM) and endoscopic submucosal dissection (ESD) are currently the two most popular methods for resecting large rectal adenomas en bloc. However, damage to the mesorectum in the case of TEM, plus the technical challenges and long procedure times of flexible ESD, are major disadvantages of these procedures. Transanal endoscopic microsurgical submucosal dissection (TEM-ESD) is a new technique, combining the ergonomic features of TEM with the minimally invasive approach of ESD. The aim of our study was to assess the feasibility and safety of TEM-ESD for resection of large rectal adenomas. We retrospectively analyzed all TEM-ESD procedures performed in Karlsruhe Municipal Hospital between 2012 and 2019, isolated all cases of adenomas, and analyzed the perioperative and follow-up data of the patients. We identified 145 cases matching our criteria. The median size of the lesions was 4.2 cm, and the median operating time was 45 min. The en bloc resection rate was 100%, and the complete en bloc resection rate was 78.6%. The overall morbidity rate was 6.9%. In a median follow-up period of 24 months, there was a local recurrence in 4.8% of the cases. TEM-ESD is a safe and feasible therapeutic option for resecting large rectal adenomas, offering high en bloc resection and low recurrence rates combined with short operating time and low morbidity. TRIAL REGISTRATION NUMBER (CLINICALTRIALS.GOV): NCT04870931.


Adherence to rehabilitation in patients with low back pain.
This study investigated adherence to low back pain rehabilitation in the clinical setting. Adherence was assessed in 105 (71 male and 34 female) patients attending private physiotherapy clinics for rehabilitation of low back pain. Three aspects of adherence were measured over the 4-week study period: attendance at clinic-based rehabilitation sessions, adherence to a home exercise component of rehabilitation, and adherence to activities and advice during clinic-based rehabilitation. Rehabilitation outcome was measured using the Patients' Measure of Perceived Rehabilitation (McDonald & Hardy 1990) and the Physiotherapists' Measure of Perceived Rehabilitation (McDonald & Hardy 1990). It was found that patients attended 87.7% of their scheduled physiotherapy rehabilitation appointments and reported completing 71.6% of their prescribed home exercises. In relation to adherence to clinic-based rehabilitation activities (as measured by the Sport Injury Rehabilitation Adherence Scale, Brewer et al. 2000), patients scored an average of 11.6/15. Although no gender differences were found, compensable patients adhered significantly less to clinic-based rehabilitation activities than did their non-compensable counterparts. Further, it was found that higher levels of adherence to clinic-based activities significantly predicted both the patients' and physiotherapists' perception of degree of rehabilitation at the end of the 4-week rehabilitation period. These findings are discussed in relation to rehabilitation strategies for physiotherapists.


Attributing social and physical meaning to ambiguous visual displays in individuals with higher-functioning autism spectrum disorders.
The weak central coherence (WCC) account of autism characterizes the learning style of individuals with this condition as favoring localized and fragmented (to the detriment of global and integrative) processing of information. This pattern of learning is thought to lead to deficits in aspects of perception (e.g., face processing), cognition, and communication (e.g., focus on disjointed details rather than "gist" or context), ultimately leading to social impairments. This study was carried out to examine whether WCC applies to social and to non-social aspects of learning alike, or, alternatively, some areas of learning (e.g., physical reasoning) are spared in autism. classic social animation as quantified in the Social Attribution Task (SAT) () and a novel animation involving physical reasoning (the Physical Attribution Task; PAT) were used to test the domain specificity of the WCC hypothesis. A pilot study involving a reference group of typically developing young adults and a group of individuals with higher-functioning autism spectrum disorders (ASDs) revealed gender differences in the reference group in regards to performance on the PAT (males outperformed females). In a follow-up case-control comparison involving only males where the ASD group was matched on age and IQ to a typically developing (TD) group of children, adolescents, and adults, the ASD group showed lower SAT scores and comparable PAT scores relative to the TD group. The interaction of diagnostic group by task was highly significant, with little overlap between the groups in the distributions of SAT minus PAT scores. These results indicated preserved integrative skills in the area of physical attribution in the ASD group, thus failing to support the WCC account as a domain-independent (or more general) model of learning in autism, while highlighting the centrality of the social deficits in the characterization of learning style in the autism spectrum disorders.


Anatomic and functional leg-length inequality: a review and recommendation for clinical decision-making. Part I, anatomic leg-length inequality: prevalence, magnitude, effects and clinical significance.
Leg-length inequality is most often divided into two groups: anatomic and functional. Part I of this review analyses data collected on anatomic leg-length inequality relative to prevalence, magnitude, effects and clinical significance. Part II examines the functional "short leg" including anatomic-functional relationships, and provides an outline for clinical decision-making. Online database--Medline, CINAHL and MANTIS--and library searches for the time frame of 1970-2005 were done using the term "leg-length inequality". Using data on leg-length inequality obtained by accurate and reliable x-ray methods, the prevalence of anatomic inequality was found to be 90%, the mean magnitude of anatomic inequality was 5.2 mm (SD 4.1). The evidence suggests that, for most people, anatomic leg-length inequality does not appear to be clinically significant until the magnitude reaches approximately 20 mm (approximately 3/4"). Anatomic leg-length inequality is near universal, but the average magnitude is small and not likely to be clinically significant.


Proteomics Analysis of Lipid Metabolism and Inflammatory Response in the Liver of Rabbits fed on a High Cholesterol Diet.
In this study, we aimed to analyze the proteomics of the liver in rabbits on a high cholesterol diet (HCD). We randomly divided New Zealand white rabbits into the normal diet group and the HCD group. We established the atherosclerosis model and measured plasma cholesterol and triglycerides. The model was successfully established using ultrasound examination and histopathological staining of the intima of aorta and liver of the two groups of rabbits. The differential proteins in the rabbit liver were analyzed using Tandem Mass Tags proteomic analysis technology. Finally, we used western blot to verify the reliability of proteomics. The results showed that compared with the control group, the serum lipid levels of rats in the HCD group was significantly increased, and the pathological sections showed the formation of atherosclerotic plaques in the aorta, inflammation, and adipose lesions in the liver. Proteomic analysis of the liver revealed 149 differences in HCD-expressed protein, which is mainly involved in inflammation and regulation of lipid and sugar metabolism. In addition, we verified differentially expressed liver proteins in the HCD group using western blot. We found that HCD caused lipid accumulation, abnormal glucose metabolism, and inflammatory response in the liver.


Rapid diagnosis of invasive pneumococcal disease in pediatric population.
The aim of this study was to evaluate the Binax NOW immunochromatographic pneumococcal antigen test for the identification of Streptococcus pneumoniae in pleural and cerebrospinal fluids from children with suspected invasive pneumococcal disease. The results were compared with those obtained by PCR. Binax NOW was applied to these samples as recommended by the manufacturer for urine and cerebrospinal samples. Detection of pneumococcal DNA was performed by real-time PCR assay targeting the autolysin gene (lytA). Of the 199 samples analyzed, 131 were positive by both Binax NOW and lytA PCR, and 36 samples were negative by both techniques. Using the real-time PCR as a comparative method to the Binax for the detection of S. pneumoniae, the sensitivity and specificity of Binax NOW was 88% and 72.5%, respectively. Of the 145 positive samples analyzed by Binax NOW, 119 showed intense coloring of the sample line and 26 showed weak intensity. Conventional culture is the most common method in clinical settings, but Binax NOW is an easier and faster test for identifying S. pneumoniae in pleural and cerebrospinal fluids from children with suspected invasive pneumococcal disease.


A novel approach to measuring response and remission in schizophrenia in clinical trials.
Pharmaceutical companies conduct clinical trials to show the efficacy and safety of new medications for the treatment of schizophrenia. After the new medications are marketed, clinicians treating patients with schizophrenia discover that a considerable number of patients do not respond to these new medications. The goals of the review are to examine the methodology and design of recent antipsychotic clinical trials, identify common flaws, and propose guidelines to fix the flaws and improve the quality of future clinical trials of antipsychotic medications. A review of recent antipsychotic clinical trials was conducted using a PubMed search. Ten recent trials published in the past four years were reviewed and their methods analyzed and critiqued. The authors identified six major methodological flaws that may explain the suboptimal response in many patients after a drug is approved. Most of the flaws are related to eligibility criteria, the misuse of the Positive and Negative Syndromes Scale (PANSS) and the lack of consensus on how to define remission, response and exacerbation in schizophrenia. Proposed guidelines for a more rigorous use of the PANSS are presented and recommendations are proposed for using uniform criteria for remission, response and exacerbation in schizophrenia. The authors recommend using standardized diagnostic interviews to screen patients for eligibility criteria and using the PANSS according to the author's recommendations and the proposed guidelines. Uniform criteria to define remission, response and exacerbation are recommended for clinical trials examining the efficacy and safety of antipsychotic drugs in schizophrenia.


Differential effects of activating D1 and D2 receptors on electrophysiology of neostriatal neurons in a rat model of Parkinson's disease induced by paraquat and maneb.
Neostriatum plays an important role in the pathophysiology of Parkinson's disease (PD). However, the changes of sensitivity of dopamine receptors of neostriatal neurons in PD have been less addressed in vivo. In the present study, systemic exposure to paraquat and maneb induced Parkinsonian symptoms and neuronal loss of substantia nigra pars compacta. Using single-unit recording methods, three types of neostriatal neurons were recorded including medium spiny-like neurons, large aspiny-like neurons and fast-spiking interneurons. In the exposed rats, increased firing activity of neostriatal neurons was revealed when compared to control rats. Following D1 receptor agonist, SKF38393 and D2 receptor agonist, LY171555 iontophoretically administrated respectively, effects of increase and decrease in firing activity were both observed in neostriatal neurons. However, stronger inhibitory effects of activating D1 receptors and weaker excitatory effects of activating D2 receptors were found in the exposed rats as compared to controls. It indicated that differential changes of sensitivity of D1 and D2 receptors in Parkinson's disease were related to the modulation of the imbalance between D1-receptor-dependent striatonigral direct pathway and D2-receptor-dependent striatopallidal indirect pathway. Our results illustrate the electrophysiological changes of in vivo neostriatal neurons in Parkinson's disease, thereby providing insight into the regulatory mechanisms of dopamine-mediated physiology.


Cardiovascular properties of LF 2.0254, a new potent vasoselective calcium channel blocker with a slow onset of action.
The effects of LF 2.0254, a new 1,4-dihydropyridine derivative, were studied in rabbit aorta stimulated by various contractile agents and in isolated guinea pig atria. LF 2.0254 inhibited in a time-dependent fashion K(+)- and CA2(+)-induced contractions of rabbit aorta with respective IC50 of 2.7 nM and 1.7 nM. An action of LF 2.0254 at the voltage operated calcium channel was consistent with the finding that LF 2.0254 antagonized 45Ca2(+)-uptake induced by depolarisation of smooth muscle, and since contractile responses evoked by (-) norepinephrine and the calcium ionophore A23187 were insensitive to the drug. In isolated paced left atria and spontaneously beating right atria of guinea pig, LF 2.0254 added for 60 min at 1 microM only slightly decreased the contractile force and beating rate. In anesthetized open-thorax dogs, LF 2.0254 (1 to 100 micrograms/kg, iv) dose-dependently lowered systemic blood pressure and increased cardiac output with a slower onset of action than nifedipine. LF 2.0254 and nifedipine decreased total peripheral, coronary, femoral and vertebral resistance. In marked contrast to nifedipine, LF 2.0254 induced only a slight decrease in left ventricular dP/dt. In conscious hypertensive dogs LF 2.0254 (0.1 and 0.3 mg/kg per os) decreased blood pressure and concomitantly increased heart rate and plasma renin activity. It is concluded that LF 2.0254 differ markedly from nifedipine in its more gradual onset of action and greater selectivity for the vasculature with respect to the myocardium.


Ontogeny of an oscillation underlying the circadian rhythm of plasma corticosterone in rats.
In previous reports (J. Physiol. 325, 493, 507, 521, 1982) we have suggested a prenatal onset of the circadian oscillation in rats. This was based mainly on the findings obtained in pups which were reared by an alternating nursing. In the present study we have applied this technique to experiments which were designed to determine the stage of gestation in the original mother that decided the phase angle of pup's hormone rhythm which was to develop later in the postnatal life. In this experiment we observed, though preliminarily, that reversal of the light-dark cycle of the pregnant mother at day 10 of pregnancy resulted in an effective reversal of pup's hormone rhythm, whereas the reversal at day 17 of pregnancy failed to affect it. From these results it is surmised that the original mother primarily determined the phase angle of the circadian oscillation in the fetus prior to the 17th day of gestation.


Stereotaxic microinjection of HSV-1 selectively decreases striatal dopamine concentrations in mice.
BALB/c mice were stereotaxically injected in the striatum with either the MP or MacIntyre strain of herpes simplex type 1. Three days later, at a time when the animals were free from overt signs of infection, they were killed by cervical dislocation and the striatum was rapidly removed. Concentrations of dopamine, norepinephrine, serotonin and their metabolites were subsequently determined by means of high-performance liquid chromatography with electrochemical detection. With both strains of virus, dopamine levels were reduced and the ratio of homovanillic acid to dopamine was elevated in MP-inoculated mice. Norepinephrine, serotonin and its metabolites were unaffected. Immunoperoxidase staining in separate, identically treated animals indicated that the infection was confined to the striatum and necrosis was minimal at this point in time. These results demonstrate that dopamine metabolism can be affected by herpes simplex in the absence of immunocytochemical evidence of infection of the cell bodies of dopaminergic neurons or cellular death.


Super Enhancer-Associated Circular RNA-CircKrt4 Regulates Hypoxic Pulmonary Artery Endothelial Cell Dysfunction in Mice.
Circular RNAs (circRNAs) have been implicated in pulmonary hypertension progression through largely unknown mechanisms. Pulmonary artery endothelial cell (PAEC) dysfunction is a hallmark in the pathogenesis of pulmonary hypertension. However, the specific role of circular RNAs in PAEC injury caused by hypoxia remains unclear. In this study, using the Western blotting, RNA pull down, Dual-luciferase reporter assay, immunohistochemistry, and immunofluorescence, we identified a novel circular RNA derived from alternative splicing of the keratin 4 gene (circKrt4). CircKrt4 was upregulated in lung tissues and plasma and specifically in PAECs under hypoxic conditions. In the nucleus, circKrt4 induces endothelial-to-mesenchymal transition by interacting with the Pura (transcriptional activator protein Pur-alpha) to promote N-cadherin gene activation. In the cytoplasm, increased circKrt4 leads to mitochondrial dysfunction by inhibiting cytoplasmic-mitochondrial shuttling of mitochondrial-bound Glpk (glycerol kinase). Intriguingly, circKrt4 was identified as a super enhancer-associated circular RNA that is transcriptionally activated by a transcription factor, CEBPA (CCAAT enhancer binding protein alpha). Furthermore, RBM25 (RNA-binding-motif protein 25) was found to regulate circKrt4 cyclization by increase the back-splicing of Krt4 gene. These findings demonstrate that a super enhancer-associated circular RNA-circKrt4 modulates PAEC injury to promote pulmonary hypertension by targeting Pura and Glpk.


Effects of physical activity and sleep duration on fertility: A systematic review and meta-analysis based on prospective cohort studies.
Subfertility is a common problem for couples in modern society. Many studies have confirmed that lifestyle factors can affect fertility although there are conflicting conclusions relating to the effects of physical activity and sleep duration on fertility. In this study, we aimed to summarize and analyze the available evidence. PubMed, Web of Science, Cochrane, and Embase databases (as of October 14, 2022) were systematically searched for eligible prospective cohort studies. Data were extracted and effect values were combined. We also performed methodological quality and bias risk assessments for all the included studies. A total of 10 eligible articles were included in our analysis; seven investigated the relationship between physical activity and fertility, and three investigated the effect of sleep duration on fertility. Compared with the lowest level of physical activity, high intensity physical activity (the highest levels of physical activity) was negatively correlated with fertility [odds ratio (OR) = 0.84; 95% confidence interval (CI): 0.70, 1.00, I 2 = 64%]. However, we did not find an association between moderate intensity physical activity and fertility (OR = 1.09; 95% CI: 0.98, 1.22, I 2 = 60%). We observed an inverse association between limited sleep duration (≤ 7 h) and fertility (OR = 0.92; 95% CI: 0.84, 1.00, I 2 = 0%) compared with 8 h of sleep. The relationship between long sleep duration (≥9 h) and fertility was not statistically significant (OR = 0.85; 95% CI: 0.60, 1.21, I 2 = 83%). According to the Newcastle-Ottawa Scale score, the overall quality of the research articles included was ranked as medium to high (6-9). Through GRADE system, the quality of evidence for the impact of high intensity physical activity and limited sleep duration on fertility was moderate, while the quality of evidence for the impact of moderate intensity physical activity and long sleep duration on fertility was low. The current evidence shows that high intensity physical activity and limited sleep time are negatively related to fertility. But there was great heterogeneity among studies, and the quality of research evidence was low to median. Thus, further high-quality research is needed to confirm this conclusion. CRD42022298137.


Impact of Lean and Fat Mass on Disease Activity and Function in Spondyloarthritis.
Changes in body mass composition, such as reduced lean and fat mass, can occur in a patient with rheumatic diseases. However, data about body composition in spondyloarthritis (SpA) showed conflicting results. The aim of the present study was to assess by DXA the distribution of lean and fat mass in SpA patients compared to healthy controls and to investigate the association between body mass composition and disease characteristics. We conducted a cross-sectional case-control study including 50 SpA patients and 50 controls. Sociodemographic data, as well as disease characteristics, were assessed. Body composition measurements and biochemical and inflammatory serum markers were evaluated. Radiographic data (Bath Ankylosing Radiologic Index and the modified Stroke Ankylosing Spondylitis Spine Score) was also recorded. No statistically significant correlation was found between the two groups regarding bone mineral density and body mass distribution. However, a lower weight was observed in the study group compared to the control group (p = 0.043). Male gender and younger age were associated with a higher lean mass (p = 0.05). C-reactive protein (CRP) level was positively correlated with lean mass (r = 0.38, p = 0.023). Similarly, BASFI was higher in patients with lower fat mass (r = -0.42, p = 0.024). A longer duration of NSAIDs intake was associated with a lower lean mass and a higher fat mass. However, no correlations were found between body mass composition and BASRI mSASSS scores as well as bone mineral density and calcium intake. Our findings suggest that younger age, male gender, and axial phenotype were associated with higher lean mass.


The gap between development and manufacturing in gene therapy: Strategic options for overcoming traps.
Gene therapy has been one of the most promising therapeutic approaches in recent years. This study analyzed a research and development (R&D) system for adeno-associated virus (AAV)-based gene therapies, and confirmed that there was a gap between the development and manufacturing capabilities. Although a start-up company that has no academic or manufacturing facilities can begin the clinical development process, it cannot successfully continue development activities without forming alliances and capital investment or, at a certain stage, without appropriate manufacturing and marketing strategies. We reviewed a series of case studies to categorize the acquisition patterns of pharmaceutical companies that are engaged in AAV gene therapy. These results provide insights into the R&D structures for AAV gene therapies from a technological management perspective.


Visualization of microvascular blood flow in mouse kidney and spleen by quantum dot injection with "in vivo cryotechnique".
The "in vivo cryotechnique" (IVCT) is a powerful tool to instantly capture blood flow, and all plasma components are well kept in tissue samples. In this study, we injected glutathione (GSH)-coated quantum dots (QDs), which emit a 650-nm-fluorescent signal with an ultraviolet excitation, into anesthetized mouse left ventricles, and IVCT was performed for kidneys, spleens and livers at 2, 5, 10, 15, 30s or 24h after the QD injection. The frozen tissues were processed to freeze-substitution fixation (FS). Then, some specimens were embedded in paraffin wax for tissue sectioning, and some were cut with a razor blade and directly mounted on glass slides. They were observed in fluorescence or confocal laser scanning microscope (CLSM). In the renal cortex, QD distribution was detected mostly in glomerular blood capillaries at 2second, and extended to peritubular blood capillaries at 5s. Distribution of horseradish peroxidase (HRP) in renal cortex at 30s after the injection was compared by the simultaneous injection with QDs. HRP was detected by a diaminobenzidin reaction in interstitium in addition to blood vessels, whereas QDs were localized only inside blood vessels. Three-dimensional reconstruction with CLSM demonstrated the capillary networks in the whole renal glomerulus. In the spleens, QDs were detected in splenic cords entering from sheathed capillaries at 10s, and extended to deeper splenic cords and also into splenic sinuses at 15s. Thus, strict time-dependent visualization of blood flow in tissue sections became possible within seconds by the new technical combination of IVCT and injection of QDs into animal organs.


Draft Genome Sequence of Acinetobacter baumannii IITR88, a Bacterium Degrading Indoles and Other Aromatic Compounds.
Here, we report the 4.16-Mb draft genome sequence of an indole-degrading bacterium, Acinetobacter baumannii IITR88, isolated from the Bhagirathi river in India. A total of 4,069 coding regions (CDSs), 3 rRNAs, and 52 tRNAs were predicted. Genes for the degradation of indoles, phenylacetaldehyde, anthranilate, and several other aromatic compounds were present.


Human glucagon and vasoactive intestinal polypeptide (VIP) stimulate free fatty acid release from human adipose tissue in vitro.
Glucagon, vasoactive intestinal polypeptide and secretin are strong stimulators of lipolysis in adipose tissue from laboratory animals. Yet, in human adipose tissue these data could not be confirmed under comparable experimental conditions. Using pH stat titration, an advanced in vitro test system for evaluating lipolysis, it was possible to demonstrate lipolytic activity for glucagon down to a concentration of 10(-8) mol/l. This is comparable to the minimal effective doses in rat adipose tissue and corresponds to the effect of equimolar concentrations of noradrenaline in man. Secretin with an amino acid sequence very similar to glucagon was not lipolytically active, while VIP stimulated free fatty acid release in a concentration of 10(-6) mol/l. Since the minimal effective dose of glucagon is only 30 times greater than the plasma levels a physiological significance of these finding may be suggested. The lipolytic activity of VIP seems to be only of pharmacological interest.


The chaperonin 60 protein SlCpn60α1 modulates photosynthesis and photorespiration in tomato.
Photosynthesis, an indispensable biological process of plants, produces organic substances for plant growth, during which photorespiration occurs to oxidize carbohydrates to achieve homeostasis. Although the molecular mechanism underlying photosynthesis and photorespiration has been widely explored, the crosstalk between the two processes remains largely unknown. In this study, we isolated and characterized a T-DNA insertion mutant of tomato (Solanum lycopersicum) named yellow leaf (yl) with yellowish leaves, retarded growth, and chloroplast collapse that hampered both photosynthesis and photorespiration. Genetic and expression analyses demonstrated that the phenotype of yl was caused by a loss-of-function mutation resulting from a single-copy T-DNA insertion in chaperonin 60α1 (SlCPN60α1). SlCPN60α1 showed high expression levels in leaves and was located in both chloroplasts and mitochondria. Silencing of SlCPN60α1using virus-induced gene silencing and RNA interference mimicked the phenotype of yl. Results of two-dimensional electrophoresis and yeast two-hybrid assays suggest that SlCPN60α1 potentially interacts with proteins that are involved in chlorophyll synthesis, photosynthetic electron transport, and the Calvin cycle, and further affect photosynthesis. Moreover, SlCPN60α1 directly interacted with serine hydroxymethyltransferase (SlSHMT1) in mitochondria, thereby regulating photorespiration in tomato. This study outlines the importance of SlCPN60α1 for both photosynthesis and photorespiration, and provides molecular insights towards plant genetic improvement.


Parameter identification in dynamical models of anaerobic waste water treatment.
Biochemical reactions can often be formulated mathematically as ordinary differential equations. In the process of modeling, the main questions that arise are concerned with structural identifiability, parameter estimation and practical identifiability. To clarify these questions and the methods how to solve them, we analyze two different second order models for anaerobic waste water treatment processes using two data sets obtained from different experimental setups. In both experiments only biogas production rate was measured which complicates the analysis considerably. We show that proving structural identifiability of the mathematical models with currently used methods fails. Therefore, we introduce a new, general method based on the asymptotic behavior of the maximum likelihood estimator to show local structural identifiability. For parameter estimation we use the multiple shooting approach which is described. Additionally we show that the Hessian matrix approach to compute confidence intervals fails in our examples while a method based on Monte Carlo Simulation works well.


The stability of NPM1 oligomers regulated by acidic disordered regions controls the quality of liquid droplets.
The nucleolus is a membrane-less nuclear body that typically forms through the process of liquid-liquid phase separation (LLPS) involving its components. NPM1 drives LLPS within the nucleolus and its oligomer formation and inter-oligomer interactions play a cooperative role in inducing LLPS. However, the molecular mechanism underlaying the regulation of liquid droplet quality formed by NPM1 remains poorly understood. In this study, we demonstrate that the N-terminal and central acidic residues within the intrinsically disordered regions (IDR) of NPM1 contribute to attenuating oligomer stability, although differences in the oligomer stability were observed only under stringent conditions. Furthermore, the impact of the IDRs is augmented by an increase in net negative charges resulting from phosphorylation within the IDRs. Significantly, we observed an increase in fluidity of liquid droplets formed by NPM1 with decreased oligomer stability. These results indicate that the difference in oligomer stability only observed biochemically under stringent conditions has a significant impact on liquid droplet quality formed by NPM1. Our findings provide new mechanistic insights into the regulation of nucleolar dynamics during the cell cycle.


MiR-19a modulates hypoxia-mediated cell proliferation and migration via repressing PTEN in human pulmonary arterial smooth muscle.
The dysfunction of human pulmonary arterial smooth muscle cells (HPASMCs) has been suggested to participate in the pathophysiology of pulmonary arterial hypertension (PAH). This study determined miR-19a expression in hypoxia-induced HPASMCs and explored the mechanistic actions of miR-19a in hypoxia-induced HPASMC proliferation and migration. QRT-PCR and western blot assays respectively determined the mRNA and protein expression of miR-19a, phosphatase and tensin homolog (PTEN) and hypoxia-inducible factor-1 alpha (HIF-1α). In vitro functional assays determined HPASMC proliferation and migration, respectively. Luciferase reporter assay determined interaction between miR-19a and PTEN. The knockdown effects of miR-19a on PAH were confirmed in in vivo mice model. Hypoxia treatment time-dependently up-regulated miR-19a expression and enhanced cell proliferation in HPASMCs. MiR-19a overexpression increased cell proliferation and migration of HPASMCs, while repression of miR-19a reduced cell proliferative and migratory potentials of hypoxia-treated HPASMCs. Bioinformatics analysis and luciferase reporter assay showed that PTEN 3' untranslated region was targeted by miR-19a, and miR-19a repressed the mRNA and protein expression of PTEN in HPASMCs. Further rescue studies revealed that miR-19a regulated proliferative and migratory potentials of hypoxia-treated HPASMCs via suppressing PTEN expression. In addition, HIF-1α was identified as one of the mediators for the hypoxia-induced aberrant expression levels of miR-19a and PTEN. MiR-19a overexpression enhanced PI3K/AKT signaling, which was attenuated by enforced expression of PTEN in HPASMCs. More importantly, knockdown of miR-19 attenuated the chronic hypoxia-induced PAH in in vivo mice model. This study presented a novel mechanistic action of miR-19a-mediated cell proliferation and migration of HPASMCs.


Differential responses of the antennal proteome of male and female migratory locusts to infection by a fungal pathogen.
The narrow host range entomopathogenic fungus, Metarhizium acridum, is an environmentally friendly acridid specific pathogen used for locust control. The locust is capable of responding within hours of infection, however, little is known concerning how the locust detects the pathogen. Here, we have identified 3213 proteins in the infected antennal proteome of the migratory locust, Locusta migratoria. iTRAQ comparative analyses of antennal proteomes identified 194 differentially abundant proteins (DAPs) between uninfected and infected males, 218 DAPs between uninfected and infected females, and 240 DAPs between infected males and infected females. In relation to olfaction, a total of 29 chemosensory proteins (CSPs), 9 odorant binding proteins (OBPs), 31 odorant receptors (ORs), and 8 ionotropic receptors (IRs) were differentially abundant after M. acridum infection, with a subset of 12 proteins found in both infected male and female antennae not present in uninfected individuals. The time course of the gene expression profiles of olfaction related DAPs were investigated by quantitative real-time PCR (qRT-PCR). Our data indicate significant changes in the antennal proteomes of male and female locusts in response to a microbial pathogen, highlighting the potential participation of olfactory processes in pathogen detection and response. BIOLOGICAL SIGNIFICANCE: The ability of an organism to detect microbial pathogens is essential for mounting a response to mitigate the spread of the infection. Using iTRAQ-based proteomic analyses changes in the protein repertoire of the antennae of male and female locusts in response to infection by a host-specific pathogen were determined. These data show proteomic alterations that are also sex-specific, identifying members of olfactory pathways that are modified in response to infection. Our data identify antennal and related olfactory proteins that are candidates for mediating host detection of pathogens, and that may contribute to subsequent behavioral and/or immune responses of the host to the infection challenge.


Scleral fixation for a phakic anterior chamber disc intraocular lens.
Angle-supported phakic anterior chamber intraocular lenses (IOLs) require a high degree of anterior vaulting, but this can cause an inadequate distance between the IOL and the cornea, especially during rubbing or squeezing of the eye. A vaultless implant was designed by anchoring 3 supple hepatics through the anterior chamber into the sclera. Thus, a suspended rather than a wedged implant was achieved. The lens was implanted in 3 eyes of 2 patients in 1990. Good tolerance of the IOL was maintained over 7 years of follow-up. A suspended anterior chamber phakic IOL maintains an adequate position with lack of bowing and shows promising results.


Chromosomes of metastatic retinoblastoma.
Chromosomal abnormalities were detected in the bone marrow of a girl with disseminated retinoblastoma. One of these abnormalities, i(6p), is characteristic of primary retinoblastoma and its presence confirmed the diagnosis of bone marrow infiltration with retinoblastoma. This approach could be extended to study other tumours of childhood.


Does fasting plasma glucose values 5.1-5.6 mmol/l in the first trimester of gestation a matter?
The aim of the study was to investigate the effect of treatment on pregnancy outcomes among women who had fasting plasma glucose (FPG) 5.1-5.6 mmol/l in the first trimester of pregnancy. We performed a secondary-analysis of a randomized community non-inferiority trial of gestational diabetes mellitus (GDM) screening. All pregnant women with FPG values range 5.1-5.6 mmol/l in the first trimester of gestation were included in the present study (n=3297) and classified to either the (i) intervention group who received treatment for GDM along with usual prenatal care (n=1,198), (ii) control group who received usual-prenatal-care (n=2,099). Macrosomia/large for gestational age (LGA) and primary cesarean-section (C-S) were considered as primary-outcomes. A modified-Poisson-regression for binary outcome data with a log link function and robust error variance was used to RR (95%CI) for the associations between GDM status and incidence of pregnancy outcomes. The mean maternal age and BMI of pregnant women in both study groups were similar. There were no statistically significant differences in the adjusted risks of adverse pregnancy outcomes, including macrosomia, primary C-S, preterm birth, hyperbilirubinemia, preeclampsia, NICU-admission, birth trauma, and LBW both groups. It is found that treating women with first-trimester FPG values of 5.1-5.6 mmol/l could not improve adverse pregnancy outcomes including macrosomia, Primary C-S, Preterm birth, hypoglycemia, hypocalcemia, preeclampsia, NICU admission, Birth trauma and LBW. Therefore, extrapolating the FPG cut-off point of the second trimester to the first -which has been proposed by the IADPSG, might therefore not be appropriate. https://www.irct.ir/trial/518, identifier IRCT138707081281N1.


Essential updates 2020/2021: Recent topics in surgery and perioperative therapy for esophageal cancer.
In this review, we focused on four topics, namely, minimally invasive esophagectomy (MIE), robot-assisted minimally invasive esophagectomy (RAMIE), conversion and salvage surgery, and neoadjuvant and adjuvant therapy, based on notable reports published in the years 2020 and 2021. It seems that while the short-term outcomes of minimally invasive Ivor Lewis esophagectomy (MIE-IL) were better than those of open Ivor Lewis esophagectomy (OE-IL), there were no significant differences in the long-term outcomes between MIE-IL and OE-IL. Similarly, the short-term outcomes of minimally invasive McKeown esophagectomy (MIE-MK) were better than those of open McKeown esophagectomy (OE-MK), while there were no significant differences in the long-term outcomes between MIE-MK and OE-MK. Furthermore, the short-term outcomes of robot-assisted minimally invasive Ivor Lewis esophagectomy (RAMIE-IL) were superior to those of completely minimally invasive Ivor Lewis esophagectomy (CMIE-IL). On the other hand, there were advantages and disadvantages in relation to the short-term outcomes of robot-assisted minimally invasive McKeown esophagectomy (RAMIE-MK) as compared with completely minimally invasive McKeown esophagectomy (CMIE-MK). However, there were no significant differences in the long-term outcomes between RAMIE-MK and CMIE-MK. Further research is needed to evaluate of short-term and long-term outcomes of transmediastinal esophagectomy with and without robotic assistance. Both induction chemotherapy and induction chemoradiotherapy appear to be promising to secure a higher rate of conversion surgery. Neoadjuvant chemoimmunotherapy and chemoimmunoradiotherapy have shown promising results and are expected as new powerful therapies.


Topographical representation in rabbit cerebellar flocculus for various afferent inputs from the brainstem investigated by means of retrograde axonal transport of horseradish peroxidase.
A small amount of horseradish peroxidase (HRP) was injected into several small areas of the flocculus and adjacent ventral paraflocculus of albino rabbits. Labeled cells were surveyed through the inferior olive, vestibular nuclear complex, nucleus reticularis tegmenti pontis, pontine nucleus and abducens nucleus. Distribution of labeled neurons in these structures varied depending on the location of injection sites, indicating the existence of a fine topographical organization for afferent inputs in the flocculus and adjacent ventral paraflocculus.


Monitoring yoctomole alkaline phosphatase by capillary electrophoresis with on-capillary catalysis-electrochemical detection.
An electrophoretically mediated microanalysis method for detection of yoctomole (ymol) alkaline phosphatase (ALP) was developed by a combination of on-capillary enzyme-catalyzed reaction and electrochemical detection. In this method, ALP molecules were electrokinetically injected into a capillary of 10 microm i.d. and then electromigrated to the section of the capillary immersed in a warm water bath of 37 degrees C, where ALP reacted for a certain time with disodium phenyl phosphate as the enzyme substrate. ALP could be measured through determining the electroactive product phenol of the enzyme-catalyzed reaction by using electrochemical detection. The phenol concentration was proportional to the mass of ALP. As a catalyst, ALP was not consumed during the reaction, which provided amplification of signal with prolonged the reaction time. In order to enhance the signal-to-noise ratio, the detection end of the capillary was etched to a horn-shape and a single carbon fiber microcylinder electrode of 6 microm in diameter as the working electrode was inserted into the detection end of the capillary. Under these conditions, the mass of ALP as low as 1.2 x 10(-22) mol (72 molecules) or 4.0 x 10(-23) mol (24 molecules) could be detected for the on-capillary reaction time of 15 min or 2h.


Phylogenetic placement of Ceratophyllum submersum based on a complete plastome sequence derived from nanopore long read sequencing data.
Eutrophication poses a mounting concern in today's world. Ceratophyllum submersum L. is one of many plants capable of living in eutrophic conditions, therefore it could play a critical role in addressing the problem of eutrophication. This study aimed to take a first genomic look at C. submersum. Sequencing of gDNA from C. submersum yielded enough reads to assemble a plastome. Subsequent annotation and phylogenetic analysis validated existing information regarding angiosperm relationships and the positioning of Ceratophylalles in a wider phylogenetic context.


N-Substituted 5-amino-1-hydroxymethyl-cyclopentanetriols: A new family of activity promotors for a GM1-gangliosidosis related human lysosomal β-galactosidase mutant.
From 1,2;3,4-di-O-isopropylidene-α-D-galactopyranose, a series of highly functionalized (hydroxymethyl)cyclopentanes was easily available. In line with reports by Reymond and Jäger on similar structures, these amine containing basic carbasugars are potent inhibitors of β-D-galactosidases and, for the first time, could be shown to act as pharmacological chaperones for GM1-gangliosidosis-associated lysosomal acid β-galactosidase mutant R201C, thus representing a new structural type of pharmacological chaperones for this lysosomal storage disease.


[Colloids versus crystalloids in objective-guided fluid therapy, systematic review and meta-analysis. Too early or too late to draw conclusions].
Several clinical trials on Goal directed fluid therapy (GDFT) were carried out, many of those using colloids in order to optimize the preload. After the decision of European Medicines Agency, there is such controversy regarding its use, benefits, and possible contribution to kidney failure. The objective of this systematic review and meta-analysis is to compare the use of last-generation colloids, derived from corn, with crystalloids, in GDFT, to determine complications and mortality associated associated. A bibliographic research was carried out in Medline, Pubmed, Embase and Cochrane Library, corroborating randomized clinical trials in those crystalloids are compared to colloids in GDFT for mayor non-cardiac surgery in adults. One hundred thirty references were found, among those 38 were selected, and 29 analyzed; of these, 6 were included for systematic review and meta-analysis, including 390 patients. It was perceived that the use of colloids it not associated with the increase of complications, but rather with a tendency to a higher mortality (RR [95% IC] 3.87 [1.121, 13, 38]); I(2)=0.0%; p=0.635). Due to this meta-analysis' limitations for small number of randomized clinical trials and patients included, the results should be taken cautiously, and it is proposed to carry out new randomized clinical trials, with enough statistical power, comparing balanced and non-balanced colloids to balanced and non-balanced crystalloids, following the protocols of GDFT, respecting current guidelines and suggestions made by groups of experts.


System constants for the bis(cyanopropylsiloxane)-co-methylsilarylene HP-88 and poly(siloxane) Rtx-440 stationary phases.
The solvation parameter model is used to characterize the retention properties of the bis(cyanopropylsiloxane)-co-methylsilarylene, HP-88, and poly(siloxane), Rtx-440, stationary phases over the temperature range 60-140 degrees C. HP-88 is among the most cohesive, dipolar/polarizable and hydrogen-bond basic of stationary phases for open-tubular column gas chromatography. It has no hydrogen-bond acidity or capacity for electron lone pair interactions. It exhibits similar selectivity to the poly(cyanopropylsiloxane) stationary phase SP-2340. Rtx-440 is a low-polarity, low-cohesion stationary phase with a moderate capacity for dipolar/polarizable and hydrogen-bond base interactions. It has no hydrogen-bond acidity and possesses weak electron lone pair interactions. It has unique selectivity when compared against a system constants database for 28 common stationary phase compositions. Cluster analysis indicated that the poly(cyanopropylphenyldimethylsiloxane) stationary phase containing 6% cyanopropylphenylsiloxane monomer, DB-1301, the poly(dimethyldiphenylsiloxane) stationary phase containing 20% diphenylsiloxane monomer, Rtx-20, the poly(siloxane) stationary phase of unknown composition, DB-624, and DX-1 [a mixture of poly(dimethylsiloxane) and poly(ethylene glycol) 9:1] are the closest selectivity matches in the database. The selectivity of DB-1301 and Rtx-440 are very similar for solutes with weak hydrogen-bond acidity allowing one stationary phase to be substituted for the other with likely success. For strong hydrogen-bond acids, such as phenols, DB-1301 and Rtx-440 exhibit different selectivity.


Indocyanine green fluorescence-navigated sentinel node biopsy showed higher sensitivity than the radioisotope or blue dye method, which may help to reduce false-negative cases in skin cancer.
Although sentinel lymph node (SLN) biopsy using radioisotope (RI) and blue dye (BD) achieved a high detection rate, approximately 5% of melanomas with negative SLNs develop nodal metastasis. We tested a new lymphatic navigation method using indocyanine green fluorescence imaging (ICG-FI) to detect such "occult" SLNs. Thirty-four skin cancer patients received SLN biopsy with the following three methods: RI (99Tc-tin colloid), BD (2% patent blue), and ICG (0.5% indocyanine green). Lymph nodes detected by any of the three methods were counted as SLNs. ICG-FI detected more SLNs in 8 out of the 34 cases (24%). The average numbers of SLNs detected by ICG-FI, RI, and BD were 2.18, 1.76, and 1.73, respectively. Interestingly, ICG-FI not only detected more SLNs in one basin (ICG-FI: 1.64, RI: 1.50, and BD: 1.51 SLNs per basin), but also detected additional SLNs in other basins (ICG-FI: 1.32, RI: 1.18, and BD: 1.15 basins per case). ICG-FI detected SLNs more efficiently than did the conventional methods, and these "occult" SLNs may offer an explanation for some false-negative cases. We recommend using ICG-FI in addition to a conventional method to reduce the risk of overlooking these "occult" SLNs.


Toxocara vitulorum intestinal impaction in male White Fulani calves: a case report from Nigeria.
Post mortem examinations were performed on the carcasses of two male White Fulani calves that reportedly died within a herd of about 45 cattle. Similar deaths were recorded in three other calves of the same age bracket (8-13 weeks) within a 5-day period about two weeks ago but no post mortem examinations were conducted on any of them unlike this present case. Contagious bovine pleuropneumonitis vaccination was reportedly up-to-date and the herd were recently treated with Diminazene aceturate (Berenil®) and Oxytetracycline® LA about two (2) months ago against haemoparasitism. Carcasses were in poor body condition with pale mucous membranes. The small intestines of both carcasses were grossly enlarged and impacted with numerous strands of round worms resulting in ballooned intestines with small ingesta and gas along with near empty large intestines containing few spots of faecal materials and gas. The worms were identified as Toxocara vitulorum while a diagnosis of T. vitulorum intestinal impaction was made. This case report highlights possible fatality that can be occasioned by these often overlooked mere worm burdens in calves within a herd. This calls for proactive measures to guard against this silent farmer's "kill joy" for meaningful and effective cattle production.


Tensile radial stress in the spinal cord related to arachnoiditis or tethering: a numerical model.
Spinal arachnoiditis comprises fibrous scarring of the subarachnoid space, following spinal trauma or inflammation, and is often associated with syringomyelia. We hypothesised that cord-to-dura attachments could cause transient tensile cord radial stress, as pressure waves propagate. This was tested in a fluid-structure interaction model, simulating three types of cord tethering, with 'arachnoiditis' confined to a short mid-section of the cord. The annular system was excited abdominally with a short transient, and the resulting Young and Lamb waves and reflections were analysed. Radial mid-section tethering was less significant than axial tethering, which gave rise to tensile radial stress locally when the cord was not fixed cranially. Simulated as inextensible string connections to the dura, arachnoiditis caused both localised tensile radial stress and localised low pressure in the cord as the transient passed. The extent of these effects was sensitive to the relative stiffness of the dura and cord. Tensile radial stress may create a syrinx in previously normal cord tissue, and transiently lowered pressure may draw in interstitial fluid, causing the syrinx to enlarge if fluid exit is inhibited. The suggested mechanism could also explain the juxtaposition of syrinxes to regions of arachnoiditis.


hCD46 receptor is not required for measles vaccine Schwarz strain replication in vivo: Type-I IFN is the species barrier in mice.
Measles virus has been successfully attenuated on chicken embryo cells to obtain a highly efficient and safe live attenuated vaccine, administered thus far to billions of children. Measles virus attenuation has long been described to involve a modification of cellular tropism with the use of human CD46 ubiquitous receptor. Nevertheless, the use of this receptor in vivo is not obvious. In this study we use four different mouse models to decipher the respective part of hCD46 receptor and type-I interferon response in measles host restriction. We observed that only type-I interferon restricts viral replication of attenuated MV Schwarz strain in mice, independently of the presence of hCD46 receptor. By comparing measles virus immunogenicity in the different models, we confirmed that there was no impact on the absence of this receptor on the immune response. Therefore, we propose to simplify the mouse model.


Biosynthesis of brush border glycoproteins by human small intestinal mucosa in organ culture.
The incorporation of [14C]glucosamine into brush border glycoproteins by human small intestinal mucosa in organ culture has been investigated. The experiments were based on the observations that (1) isolated brush border membrane fragments from cultured explants showed an unchanged pattern of protein bands and brush border enzyme activities on sodium dodecyl sulfate/polyacrylamide gels after electrophoresis and (2) the rate of overall [14C]glucosamine incorporation measured in the tissue homogenate remained constant up to 48 h. After 24 h of culture, the radioactivity peaks on gels due to incorporation of [14C]glucosamine were found exclusively in the high molecular weight region and corresponded to protein bands identified as maltase-glucoamylase, lactase, sucrase-isomaltase, enterokinase and alkaline phosphatase. Enzymatic activity could not be assigned to the three remaining labelled bands. Most of these glycoproteins were already labelled after 5 h. Newly glycosylated brush border enzymes remained predominantly associated with the brush border membrane of intact cells with little release into the medium up to 24 h.


Combinatorial triple-selective labeling as a tool to assist membrane protein backbone resonance assignment.
Obtaining NMR assignments for slowly tumbling molecules such as detergent-solubilized membrane proteins is often compromised by low sensitivity as well as spectral overlap. Both problems can be addressed by amino-acid specific isotope labeling in conjunction with (15)N-(1)H correlation experiments. In this work an extended combinatorial selective in vitro labeling scheme is proposed that seeks to reduce the number of samples required for assignment. Including three different species of amino acids in each sample, (15)N, 1-(13)C, and fully (13)C/(15)N labeled, permits identification of more amino acid types and sequential pairs than would be possible with previously published combinatorial methods. The new protocol involves recording of up to five 2D triple-resonance experiments to distinguish the various isotopomeric dipeptide species. The pattern of backbone NH cross peaks in this series of spectra adds a new dimension to the combinatorial grid, which otherwise mostly relies on comparison of [(15)N, (1)H]-HSQC and possibly 2D HN(CO) spectra of samples with different labeled amino acid compositions. Application to two α-helical membrane proteins shows that using no more than three samples information can be accumulated such that backbone assignments can be completed solely based on 3D HNCA/HN(CO)CA experiments. Alternatively, in the case of severe signal overlap in certain regions of the standard suite of triple-resonance spectra acquired on uniformly labeled protein, or missing signals due to a lack of efficiency of 3D experiments, the remaining gaps can be filled.


Voltage-independent catecholamine release mediated by the activation of muscarinic receptors in guinea-pig adrenal glands.
1. The differences between the mechanisms of muscarinic and nicotinic receptor-mediated catecholamine secretion with respect to their dependence on voltage changes and extracellular Ca were examined using perfused adrenal glands of the guinea-pig. 2. Acetylcholine (ACh, 10(-6) to 10(-3) M) caused a dose-dependent increase in catecholamine secretion. The ED50 value for ACh was 7 x 10(-5) M. In the presence of atropine (10(-5) M), the dose-response curve for ACh was shifted to the right. Hexamethonium (5 x 10(-4) M) preferentially reduced the responses to higher concentrations of ACh (greater than 10(-5) M). Pilocarpine (5 x 10(-4) M) and nicotine (3 x 10(-5) M) also stimulated catecholamine release. 3. During perfusion with isotonic KCl solution, ACh and pilocarpine, but not nicotine, evoked catecholamine secretion. These responses were abolished by atropine (10(-6) M). Pilocarpine-stimulated catecholamine secretion was enhanced during perfusion with isotonic KCl solution. Under these conditions, hexamethonium (10(-3) M) significantly augmented ACh-evoked catecholamine release. 4. During perfusion with either Ca-free isotonic KCl or Ca-free Locke solution, ACh and pilocarpine caused a partial increase in catecholamine secretion whereas nicotine and high K solution (56 mM) did not. The responses to ACh and pilocarpine were completely inhibited by atropine but not by hexamethonium. 5. When guinea-pig adrenal glands were perfused with isotonic KCl solution containing 2.2 mM Ca which was subsequently removed and replaced with EGTA, ACh-induced catecholamine secretion was similar in magnitude to that observed during perfusion with Locke solution. 6. We conclude that both nicotinic and muscarinic receptors are involved in ACh-induced catecholamine secretion from guinea-pig adrenal chromaffin cells. Activation of muscarinic or nicotinic receptors appears to stimulate catecholamine release through different mechanisms with respect to both voltage-dependence and Ca requirements.


Mutational Signature and Transcriptomic Classification Analyses as the Decisive Diagnostic Tools for a Cancer of Unknown Primary.
Cancer of unknown primary is a group of metastatic tumors in which the standard diagnostic workup fails to identify the site of origin of the tumor. The potential impact of precision oncology on this group of patients is large, because actionable driver mutations and a correct diagnosis could provide treatment options otherwise not available for patients with these fatal cancers. This study investigated if comprehensive genomic analyses could provide information on the origin of the tumor. Here we describe a patient whose tumor was misdiagnosed at least three times. Next-generation sequencing, a patient-derived xenograft mouse model, and bioinformatics were used to identify an actionable mutation, predict resistance development to the targeted therapy, and correctly diagnose the origin of the tumor. Transcriptomic classification was benchmarked using The Cancer Genome Atlas (TCGA). Despite the lack of a known primary tumor site and the absence of diagnostic immunohistochemical markers, the origin of the patient's tumor was established using the novel bioinformatic workflow. This included a mutational signature analysis of the sequenced metastases and comparison of their transcriptomic profiles to a pan-cancer panel of tumors from TCGA. We further discuss the strengths and limitations of the latter approaches in the context of three potentially incorrectly diagnosed TCGA lung tumors. Comprehensive genomic analyses can provide information on the origin of tumors in patients with cancer of unknown primary.


The effect of lymphadenectomy on survival and recurrence in patients with ovarian cancer: a systematic review and meta-analysis.
Our objective was to perform a meta-analysis examining the effectiveness of lymphadenectomy in patients with ovarian cancer. PubMed and CENTRAL databases were searched on 15 November 2015 using the terms 'lymphadenectomy', 'ovarian cancer', 'dissection', 'para-aortic', 'pelvic' and survival. Prospective and retrospective studies comparing the outcomes of surgery with or without lymphadenectomy were included. Outcomes were 5-year overall survival, progression-free survival and recurrence rate. Of the 556 studies identified, 3 randomized controlled trials and 11 retrospective studies were included. Lymphadenectomy was associated with greater 5-year overall survival than no lymphadenectomy (pooled odds ratio = 1.58, 95% confidence interval: 1.41-1.77, p < 0.001). There was no difference in progression-free survival between the groups (pooled overall survival = 1.62, 95% confidence interval: 0.82-3.21, p = 0.168). Lymphadenectomy was associated with greater progression-free survival in randomized clinical trials (pooled overall survival = 1.57, 95% confidence interval: 1.11-2.21, p = 0.010), but not in retrospective studies. Lymphadenectomy was associated with a significantly lower recurrence rate (pooled overall survival = 0.51, 95% confidence interval: 0.30-0.85, p = 0.011). Lymphadenectomy was associated with greater 5-year overall survival in patients with both early and advanced stage cancer, but was associated with greater progression-free survival and lower recurrence rate only in patients with advanced stage cancer. Lymphadenectomy is associated with greater 5-year overall survival in patients with early and advanced stage ovarian cancer, but an effect on progression-free survival and recurrence rate was only found in patients with advanced stage ovarian cancer.


The caregiver reaction assessment: psychometrics, and temporal stability in primary caregivers of Norwegian cancer patients in late palliative phase.
Limited research has been done on the situation of primary caregivers (PCs) to patients staying at home with cancer in the palliative phase. This study uses the Caregiver Reaction Assessment (CRA) to examine PCs caregiver burden and well-being. The sample consisted of 85 PCs examined at baseline, among them 42 were re-examined four months later. We explored the psychometric properties of the CRA at baseline, and introduced a CRA sum score. The CRA sum score was correlated with the Short Form 36 (SF-36) and The Hospital Anxiety and Depression Scale (HADS) scores. The internal consistency of the CRA dimensions varied between alpha 0.57 and 0.85, and the factor structure was in line with earlier studies. The CRA sum score correlated significantly with all mental dimensions on the SF-36 and the HADS. At baseline the PCs showed significantly worse scores except for family support when compared to newly diagnosed cancer patients. The mean scores on the CRA dimensions as well as the total score did not change significantly from baseline to follow-up. We have confirmed the psychometric properties of the CRA, which seems to be an appropriate instrument for assessment the PCs caregiver situation.


Hypothesis on microangiopathy of cutaneous capillaries.
Microangiopathy is a more or less pronounced PAS deposit-located exterior to the endothelial cells of the lymphatics and the basal membrane of the capillaries. This lesion, found in various normal and pathological states, has generated numerous pathogenic hypotheses. The presence of microangiopathy in 5 groups of 50 subjects representing five different clinical conditions, subjects over 60 years old or less than 40, diabetics, latent diabetics or patients with severe coronary heart disease, together with microscopic and/or ultrastructural lesions of the connective tissue (fibroblasts, collagen and elastic fibers, ground substance) has enabled us to propose a pathogenic hypothesis applicable to any microangiopathy. The initial change, hereditary or acquired, would be fibroblastic or interstitial. It would be characterized by the accumulation of glycoproteins, proteoglyacans and soluble collagen in the interstitium. Incomplete drainage of these macromolecules would occur around the blood and lymphatic capillaries and manifest itself by a PAS deposit, the hallmark of the microangiopathy.


Modified siRNA effectively silence inducible immunoproteasome subunits in NSO cells.
The pathogenesis of autoimmune and neurodegenerative diseases involves overexpression of inducible subunits of the immunoproteasome. However, the clinical application of inhibitors to inducible subunits of the immunoproteasome has been limited due to systemic toxicity. Here, we designed siRNAs that efficiently silence LMP2, LMP7 and MECL-1 gene expression. Inducible subunits of the immunoproteasome are complex siRNA targets because they have a long half-life; therefore, we introduced 2'-O-methyl modifications into nuclease-sensitive sites. This led to 90-95% silencing efficiency and prolonged silencing, eliminating the need for multiple transfections. Furthermore, we showed that in the absence of transfection reagent, siRNAs with lipophilic residues were able to penetrate cells more effectively and decrease the expression of inducible immunoproteasome subunits by 35% after 5 days. These results show that siRNA targeted to inducible immunoproteasome subunits have great potential for the development of novel therapeutics for autoimmune and neurodegenerative diseases.


Syntheses of the methyl glycosides of the repeating units of chondroitin 4- and 6-sulfate.
3,4,6-Tri-O-acetyl-D-galactal was transformed into methyl 6-O-acetyl-2-azido-4-O-benzyl-2-deoxy-beta-D-galactopyranoside and its 4-O-acetyl-6-O-benzyl analogue, each of which was glycosylated with activated, O-acetylated derivatives of methyl D-glucopyranosyluronate. The resulting beta-(1----3)-linked disaccharide derivatives were each reductively N-acetylated, hydrogenolysed, O-sulfated, and saponified to afford the disodium salts of methyl 2-acetamido-2-deoxy-3-O-(beta-D-glucopyranosyluronic acid)-4-O-sulfo-beta-D-galactopyranoside and the 6-O-sulfo analogue. D-Galactal was also transformed into activated derivatives of 2-azido-3,6-di-O-benzyl-2-deoxy-D-galactopyranose and their 3,4-di-O-benzyl analogues with various substituents at O-4 and O-6. These glycosyl donors were condensed with 6-O-protected derivatives of methyl 2,3-di-O-benzyl-beta-D-glucopyranoside to give the beta-(1----4)-linked disaccharide derivatives, which were selectively deprotected, then oxidised at C-6 of the gluco unit, reductively N-acetylated, selectively deprotected, O-sulfated at C-4 or C-6 of the galacto unit, and hydrogenolysed to give the disodium salts of methyl 4-O-(2-acetamido-2-deoxy-4-O-sulfo-beta-D-galactopyranosyl)-beta-D- glucopyranosiduronic acid and the 6-O-sulfo analogue.


Systematic review of guidance for the collection and use of patient-reported outcomes in real-world evidence generation to support regulation, reimbursement and health policy.
Real-world evidence (RWE) plays an increasingly important role within global regulatory and reimbursement processes. RWE generation can be enhanced by the collection and use of patient-reported outcomes (PROs), which can provide valuable information on the effectiveness, safety, and tolerability of health interventions from the patient perspective. This systematic review aims to examine and summarise the available PRO-specific recommendations and guidance for RWE generation. Medical Literature Analysis and Retrieval System Online, Excerpta Medica Database, and websites of selected organisations were systematically searched to identify relevant publications. 1,249 articles were screened of which 7 papers met the eligibility criteria and were included in the review. The included publications provided PRO-specific recommendations to facilitate the use of PROs for RWE generation and these were extracted and grouped into eight major categories. These included: (1) instrument selection, (2) participation and engagement, (3) burden to health care professionals and patients, (4) stakeholder collaboration, (5) education and training, (6) PRO implementation process, (7) data collection and management, and (8) data analysis and presentation of results. The main limitation of the study was the potential exclusion of relevant publications, due to poor indexing of the databases and websites searched. PROs may provide valuable and crucial patient input in RWE generation. Whilst valuable insights can be gained from guidance for use of PROs in clinical care, there is a lack of international guidance specific to RWE generation in the context of use for regulatory decision-making, reimbursement, and health policy. Clear and appropriate evidence-based guidance is required to maximise the potential benefits of implementing PROs for RWE generation. Unique aspects between PRO guidance for clinical care and other purposes should be differentiated. The needs of various stakeholder groups (including patients, health care professionals, regulators, payers, and industry) should be considered when developing future guidelines.


Feces Metagenomes and Metagenome-Assembled Genome Sequences from Two Separate Dogs (Canis lupus familiaris) with Multiple Diarrheal Episodes.
This study reports two feces metagenomes (D84 and D85) and six metagenome-assembled genomes (MAGs). The assembled MAGs include Pseudomonas sp. strain NID84 and Acinetobacter sp. strain N2D84 from D84 and Enterococcus sp. strain N4D85, Enterococcus sp. strain N5D85, Lactobacillus sp. strain N6D85, and Leuconostoc sp. strain N7D85 from D85. Acinetobacter sp. N2D84 was identified as a human pathogen with a probability of 92%.


Comparison of Drug-Coated Balloons vs Bare-Metal Stents in Patients With Femoropopliteal Arterial Disease.
Endovascular treatment of femoropopliteal artery disease has shifted toward drug-coated balloons (DCB). However, limited data are available regarding the safety and efficacy of DCB vs bare-metal stents (BMS). The purpose of this study was to compare DCB vs BMS outcomes in a propensity-adjusted, pooled analysis of 4 prospective, multicenter trials. Patient-level data were pooled from 4 prospective, multicenter studies: the IN.PACT SFA I/II and IN.PACT SFA Japan randomized controlled DCB trials and the Complete SE and DURABILITY II single-arm BMS studies. Outcomes were compared using inverse probability of treatment weighting (IPTW). Clinical endpoints were 12-month primary patency, freedom from 36-month clinically driven target lesion revascularization, and cumulative 36-month major adverse events (MAE). The primary analysis included 771 patients (288 DCB, 483 BMS). IPTW-adjusted demographic, baseline lesion, and procedural characteristics were matched between groups. The adjusted mean lesion length was 8.1 ± 4.7 cm DCB and 7.9 ± 4.5 cm BMS. The IPTW-adjusted Kaplan-Meier estimates of 12-month primary patency (90.4% DCB, 80.9% BMS, P = 0.007), freedom from 36-month clinically driven target lesion revascularization (85.6% DCB, 73.7% BMS, P = 0.001), and cumulative incidence of 36-month MAE (25.3% DCB, 38.8% BMS, P < 0.001) favored DCB. There were no statistically significant differences observed in all-cause mortality, target limb major amputation, or thrombosis through 36 months. In a patient-level, IPTW-adjusted pooled analysis of prospective, multicenter pivotal studies, DCB demonstrated significantly higher patency, lower revascularization and MAE rates, and no statistically significant differences in mortality, amputation, or thrombosis vs BMS. This analysis supports DCB use vs BMS in moderately complex femoropopliteal lesions amenable to both treatments.


Conditioned social preference and reward value of activating oxytocin-receptor-expressing ventral tegmental area neurons following repeated daily binge ethanol intake.
Individuals with alcohol use disorder (AUD) exhibit a disruption of social behavior and dysregulation of oxytocin signaling in the brain, possibly reflecting decreased activation of oxytocin receptors (OxTRs) in reward pathways in response to social stimuli. We hypothesize that daily binge ethanol intake causes a deficit in social reward and oxytocin signaling in the ventral tegmental area (VTA). After 9 weeks of daily binge ethanol intake (blood ethanol concentration >80 mg%), OxTR-cre mice underwent conditioned place preference for social reward. Separate groups of mice were tested for the effects of binge ethanol on voluntary social interactions, food reward, locomotion, and anxiety-like behaviors. A subset of mice underwent transfection of OxTR-expressing VTA neurons (VTAOxtr ) with a light-sensitive opsin, followed by operant training to respond to light delivered to VTA. Ethanol-naïve male mice increased the time spent on the side previously paired with novel mice while ethanol-treated mice did not. Binge ethanol did not affect conditioned place preference for food reward in males, but this response was weakened in ethanol-treated females. Ethanol treatment also caused a sex-specific impairment of voluntary social interactions with novel mice. There were minimal differences between groups in measures of anxiety and locomotion. Ethanol-naïve mice had significantly greater operant responding for activation of VTAOxtr than sham-transfected mice but ethanol-treated mice did not. There was no difference in the number of VTAOxtr after binge ethanol. Daily binge ethanol causes social reward deficits that cannot be explained by nonspecific effects on other behaviors, at least in males. Only ethanol-naïve mice exhibited positive reinforcement caused by activation of VTAOxtr while daily binge ethanol did not alter the number of VTAOxtr in either males or females. Thus, subtle dysregulation of VTAOxtr function may be related to the social reward deficits caused by daily binge ethanol.


Solution NMR investigations of integral membrane proteins: Challenges and innovations.
Compared to soluble protein counterparts, the understanding of membrane protein stability, solvent interactions, and function are not as well understood. Recent advancements in labeling, expression, and stabilization of membrane proteins have enabled solution nuclear magnetic resonance spectroscopy to investigate membrane protein conformational states, ligand binding, lipid interactions, stability, and folding. This review highlights these advancements and new understandings and provides examples of recent applications.


IFITM3 and type I interferons are important for the control of influenza A virus replication in murine macrophages.
Abortive infection of macrophages serves as a "dead end" for most seasonal influenza A virus (IAV) strains, and it is likely to contribute to effective host defence. Interferon (IFN)-induced transmembrane protein 3 (IFITM3) restricts the early stages of IAV replication in epithelial cells, but IFITM3 restriction of IAV replication in macrophages has not been previously investigated. Herein, macrophages isolated from IFITM3-deficient mice were more susceptible to initial IAV infection, but late-stage viral replication was still controlled through abortive infection. Strikingly, IFNα/β receptor (IFNAR)-deficient macrophages infected with IAV were not only more susceptible to initial infection, but these cells also supported productive viral replication. Significantly, we have established that abortive IAV infection in macrophages is controlled through a type I IFN-dependent mechanism, where late-stage IAV replication can proceed in the absence of type I IFN responses. These findings provide novel mechanistic insight into macrophage-specific processes that potently shut down IAV replication.