Role of mast cells in experimental anti-glomerular basement membrane glomerulonephritis.
Recently, divergent reports on the role of mast cells (MC) in different glomerular diseases have brought our attention to their role in an accelerated model of anti-glomerular basement membrane (GBM) glomerulonephritis (GN). Genetically MC-deficient Kit(W)/Kit(W-v) mice, MC-reconstituted Kit(W)/Kit(W-v) mice and Kit+/+ control mice were subjected to anti-GBM GN. Kit(+/+) mice developed moderate proteinuria and glomerular damage following the induction of anti-GBM nephritis. In contrast, proteinuria and glomerular damage were dramatically increased in MC-deficient Kit(W)/Kit(W-v) mice. MC-reconstituted Kit(W)/Kit(W-v) mice showed proteinuria and glomerular damage comparable to Kit+/+ mice. A significant increase in infiltrating T cells and macrophages was detected in MC-deficient Kit(W)/Kit(W-v) mice as compared to Kit+/+ control mice and MC-reconstituted Kit(W)/Kit(W-v) mice. Accordingly, we observed an increase of TGF-beta1 mRNA in kidneys from Kit(W)/Kit(W-v) mice. Interestingly, we did not detect MC in the kidney using either Giemsa staining or RT-real-time PCR, but MC were found in the regional lymph nodes. Finally, mortality of Kit(W)/Kit(W-v) mice was significantly increased after the induction of anti-GBM GN due to uremia. Our report provides the first direct evidence that MC are protective in anti-GBM GN, possibly by modulating the influx of effector T cells and macrophages to inflammatory sites in the kidney.


Drug-like biimidazole derivatives dually target c-MYC/BCL-2 G-quadruplexes and inhibit acute myeloid leukemia.
Chemotherapy is the main approach for treating acute myeloid leukemia (AML). However, this therapy can cause severe side effects as well as drug resistance, hence calling for new therapeutic strategies. As c-MYC and BCL-2 are often overexpressed in AML, and synergism between c-MYC and BCL-2 promotes tumorigenesis, therefore, dual targeting of c-MYC/BCL-2 promoter G-quadruplexes (G4s) and then inhibiting the targeted gene expression would be a potential strategy in ALM treatment. In this work, in the search of dual ligands, we performed a screening assay with an in-house, imidazole-based compound library. Consequently, two drug-like biimidazole derivatives were identified as selective c-MYC/BCL-2 G4 binders, of which, BIM-2 was selected as the candidate for inhibiting AML cell growth. Then, BIM-2 was demonstrated to downregulate both c-MYC and BCL-2 expression, and thereby cause cell cycle arrest at G0/G1 phase and apoptosis in AML cells. Furthermore, the possible end-stacking binding modes between BIM-2 and c-MYC/BCL-2 G4s were revealed by NMR and molecular docking studies. Accordingly, this study provides a new class of drug-like dual-selective c-MYC/BCL-2 G4 ligands for the potential treatment of AML.


Repetitive transcranial magnetic stimulation can improve the fixation of eyes rather than the fixation preference in children with autism spectrum disorder.
Transcranial magnetic stimulation (TMS) has been introduced into the intervention of autism spectrum disorders (ASD) as a possible new therapeutic option for modifying pathological neuroplasticity. However, the stimulating protocols of rTMS for ASD have not been approved unanimously, which affects the clinical popularization and application of rTMS. In addition, there is little research on the improvement of social processing of autistic children by rTMS. We explored the clinical efficacy of rTMS and improvement of face processing with the protocol of left high-frequency and right low-frequency on bilateral dorsolateral prefrontal cortex (DLPFC), with a sample of 45 ASD participants aged 2-18. Our results showed that both the score on the Childhood Autism Rating Scale (CARS) and the fixations on the eyes of the human faces improved by two-session rTMS intervention, except for the percentage of eyes fixation. The mediation analysis indicated the item of "Adaptation to Change" of CARS mediated dominantly the improvement of eye-gaze behavior of ASD participants by rTMS. Our study revealed the mechanism of rTMS in improving the eye-gaze behavior of the autism population, deepened the understanding of the function of rTMS in treating autistic social disorders, and provided a reference for combined treatment for ASD.


One-Pot Synthesis of W2C/WS2 Hybrid Nanostructures for Improved Hydrogen Evolution Reactions and Supercapacitors.
Tungsten sulfide (WS2) and tungsten carbide (W2C) are materialized as the auspicious candidates for various electrochemical applications, owing to their plentiful active edge sites and better conductivity. In this work, the integration of W2C and WS2 was performed by using a simple chemical reaction to form W2C/WS2 hybrid as a proficient electrode for hydrogen evolution and supercapacitors. For the first time, a W2C/WS2 hybrid was engaged as a supercapacitor electrode and explored an incredible specific capacitance of ~1018 F g-1 at 1 A g-1 with the outstanding robustness. Furthermore, the constructed symmetric supercapacitor using W2C/WS2 possessed an energy density of 45.5 Wh kg-1 at 0.5 kW kg-1 power density. For hydrogen evolution, the W2C/WS2 hybrid produced the low overpotentials of 133 and 105 mV at 10 mA cm-2 with the small Tafel slopes of 70 and 84 mV dec-1 in acidic and alkaline media, respectively, proving their outstanding interfaced electrocatalytic characteristics. The engineered W2C/WS2-based electrode offered the high-performance for electrochemical energy applications.


Ring-testing and field-validation of a terrestrial model ecosystem (TME)--an instrument for testing potentially harmful substances: effects of carbendazim on earthworms.
The effects of the fungicide carbendazim (applied in the formulation Derosal) on earthworms (Lumbricidae) was determined in Terrestrial Model Ecosystem (TME) tests and field-validation studies. TMEs consisted of intact soil columns (diameter 17.5 cm; length 40 cm) taken from a grassland or, in one case, from an arable site. The TMEs were taken from the same site where the respective field-validation study was performed. The tests were performed in Amsterdam (The Netherlands), Bangor (Wales, UK), Coimbra (Portugal) and Flörsheim (Germany). The sites selected had an earthworm coenosis representative of the different land use types and regions. In addition, the differences between the coenosis found in the TMEs and the respective field sites were in general low. A high variability was found between the replicate samples, which reduces the probability of determining significant differences by the statistical evaluation of the data. Similar effects of the chemical treatment were observed on abundance as well as on biomass. Effects were most pronounced 16 weeks after application of the test chemical. The observed effects on earthworm abundance and biomass did not differ between the TME tests and the respective field-validation studies. Effects on earthworm diversity were difficult to assess since the number of individuals per species was low in general. However, the genus Lumbricus and in particular L. terrestris and L. rubellus seemed to be more affected by the chemical treatment than others. NOEC and EC50-values derived from the TME pre-test, the TME ring-test and the field-validation study indicate that the TMEs of the different partners delivered comparable results although different soils were used. Due to the high variability NOECs could often not be determined. The EC50-values for the effect of carbendazim on earthworm abundance ranged between 2.04 and 48.8 kg a.i./ha (2.71-65.2 mg/kg soil) and on earthworm biomass from 1.02 to 34.6 kg a.i./ha (1.36-46.0 mg/kg soil). These results indicate that the abundance and biomass of earthworms are suitable endpoints in ecotoxicological studies with TMEs.


Rac1 GTPase acts downstream of αPS1βPS integrin to control collective migration and lumen size in the Drosophila salivary gland.
During collective migration of the Drosophila embryonic salivary gland, the distal gland cells mediate integrin-based contacts with surrounding tissues while proximal gland cells change shape and rearrange. Here, we show that αPS1βPS integrin controls salivary gland migration through Rac1 GTPase which downregulates E-cadherin in proximal and distal gland cells, and promotes extension of actin-rich basal membrane protrusions in the distal cells. In embryos mutant for multiple edematous wings (mew), which encodes the αPS1 subunit of the αPS1βPS integrin heterodimer, or rac1 and rac2 GTPases, salivary gland cells failed to migrate, to downregulate E-cadherin and to extend basal membrane protrusions. Selective inhibition of Rac1 in just the proximal or distal gland cells demonstrate that proximal gland cells play an active role in the collective migration of the whole gland and that continued migration of the distal cells depends on the proximal cells. Loss of rac1rac2 also affected gland lumen length and width whereas, loss of mew affected lumen length only. Activation of rac1 in mew mutant embryos significantly rescued the gland migration, lumen length and basal membrane protrusion defects and partially rescued the E-cadherin defects. Independent of mew, Rac regulates cell shape change and rearrangement in the proximal gland, which is important for migration and lumen width. Our studies shed novel insight into a Rac1-mediated link between integrin and cadherin adhesion proteins in vivo, control of lumen length and width and how activities of proximal and distal gland cells are coordinated to result in the collective migration of the entire salivary gland.


Acute angle-closure glaucoma due to iris transfixation of a subluxated posterior chamber intraocular lens-capsular bag complex.
An 80-year-old woman with pseudoexfoliation syndrome developed subluxation of a posterior chamber intraocular lens (IOL)-capsular bag complex due to zonular rupture. She underwent a limited 25-gauge pars plana vitrectomy and iris transfixation of the subluxated IOL-capsule complex. The postoperative course was uneventful until the patient developed acute angle-closure glaucoma 3 weeks postoperatively. Ultrasound biomicroscopy confirmed pupillary block by the IOL-capsule complex. The patient was treated with intraocular pressure-lowering medications and a vitreous tap before a laser peripheral iridotomy could be performed successfully. The vision returned to normal, and the angle closure resolved. We recommend that a prophylactic surgical iridectomy be performed in patients with iris transfixation of a posterior chamber IOL-capsular bag complex.


Intermediates and enzymes between alpha-ketoarginine and gamma-guanidinobutyrate in the L-arginine catabolic pathway of Pseudomonas putida.
In Pseudomonas putida P2 grown on L-arginine as the sole source of carbon and nitrogen, catabolism of L-arginine forms of alpha-ketoarginine, gamma-guanidinobutyrate, and gamma-aminobutyrate. A previously undetected intermediate, gamma-guanidinobutyraldehyde, is identified as the product of alpha-ketoarginine decarboxylase. An 86-fold purification of this enzyme is described. Activity is thiamine pyrophosphate-dependent and cofactor reassociation is facilitated by divalent cations. The order of effectiveness is Mn-2+ greater than Mg-2+, Co-2+ greater than Ca-2+ greater than Ni-2+ greater than Zn-2+. An inducible enzyme that catalyzes conversion of gamma-guanidinobutyraldehyde to gamma-guanidinobutyrate has been studied in cell-free extracts. NAD-+, but no other cofactors, is required. By differential nutritional growth experiments, 4 regulatory units for the L-arginine pathway are proposed and inducers of 2 units are identified.


Facile preparation of multi-functionalized hybrid monoliths via two-step photo-initiated reactions for two-dimensional liquid chromatography-mass spectrometry.
A facile approach was developed to prepare hybrid monoliths with different functions via two-step photo-initiated reactions. Firstly, acrylopropyl polyhedral oligomertic silsesquioxane (acryl-POSS) and propargyl acrylate (PA) were used as precursors to synthesize alkynyl-functionalized hybrid monoliths via photo-initiated free radical polymerization. Secondly, the hybrid monoliths were modified with 1-octadecanethiol (ODT) and sodium 3-mercapto-1-propanesulfonate (SMPS) via photo-initiated thiol-yne click reaction to prepare reversed-phase (RP) and strong cation-exchange (SCX) hybrid monoliths, respectively. The results of chromatographic characterization indicated that the column efficiencies for alkylbenzenes on ODT-modified hybrid monolith reached 84,000-87,700 plates per meter at the velocity of 0.58mm/s, and also revealed a retention-independent efficient performance of small molecules in isocratic elution. The SMPS-modified hybrid monolith exhibited both hydrophobicity and ion-exchange mechanisms, and the dynamic binding capacity was calculated to be 1.4×10-4μmol/cm. Human Hela cells tryptic digest was well separated on ODT-modified hybrid monolith in one-dimensional RPLC-MS/MS, and 2786 unique peptides and 685 proteins were identified. Furthermore, the SMPS-modified monolith coupled with ODT-modified monolith was used for two-dimensional separation of human Hela cells tryptic digest in SCX-RPLC-MS/MS, and the results showed that 9744 unique peptides and 2749 proteins were identified. Compared to those identified in one-dimensional RP system, the total numbers of unique peptides and proteins identified in SCX-RP system increased by 249.7% and 301.3%, respectively.


Object extraction via deep learning-based marker-free tracking framework of surgical instruments for laparoscope-holder robots.
The surgical instrument tracking framework, especially the marker-free surgical instrument tracking framework, is the key to visual servoing which is applied to achieve active control for laparoscope-holder robots. This paper presented a marker-free surgical instrument tracking framework based on object extraction via deep learning (DL). The surgical instrument joint was defined as the tracking point. Using DL, a segmentation model was trained to extract the end-effector and shaft portions of the surgical instrument in real time. The extracted object was transformed into a distance image by Euclidean Distance Transformation. Next, the points with the maximal pixel value in the two portions were defined as their central points, respectively, and the intersection point of the line connecting the two central points and the plane connecting the two portions was determined as the tracking point. Finally, the object could be fast extracted using the masking method, and the tracking point was fast located frame-by-frame in a laparoscopic video to achieve tracking of surgical instrument. The proposed object extraction via a DL-based marker-free tracking framework was compared with a marker-free tracking-by-detection framework via DL. Using seven in vivo laparoscopic videos for experiments, the mean tracking success rate was 100%. The mean tracking accuracy was (3.9 ± 2.4, 4.0 ± 2.5) pixels measured in u and v coordinates of a frame, and the mean tracking speed was 15 fps. Compared to the reported mean tracking accuracy of a marker-free tracking-by-detection framework via DL, the mean tracking accuracy of our proposed tracking framework was improved by 37% and 23%, respectively. Accurate and fast tracking of marker-free surgical instruments could be achieved in in vivo laparoscopic videos by using our proposed object extraction via DL-based marker-free tracking framework. This work provided important guiding significance for the application of laparoscope-holder robots in laparoscopic surgeries.


Rapid analysis of six phthalate esters in wine by ultrasound-vortex-assisted dispersive liquid-liquid micro-extraction coupled with gas chromatography-flame ionization detector or gas chromatography-ion trap mass spectrometry.
An Ultrasound-Vortex-Assisted Dispersive Liquid-Liquid Micro-Extraction (USVADLLME) procedure coupled with Gas Chromatography-Flame Ionization Detector (GC-FID) or Gas Chromatography-Ion Trap Mass Spectrometry (GC-IT/MS) is proposed for rapid analysis of six phthalate esters in hydroalcoholic beverages (alcohol by volume, alc vol(-1), ≤40%). Under optimal conditions, the enrichment factor of the six analytes ranges from 220- to 300-fold and the recovery from 85% to 100.5%. The limit of detection (LOD) and limit of quantification (LOQ) are ≥0.022 μg L(-1) and ≥0.075 μg L(-1), respectively. Intra-day and inter-day precisions expressed as relative standard deviation (RSD), are ≤8.2% and ≤7.0%, respectively. The whole proposed methodology has demonstrated to be simple, reproducible and sensible for the determination of trace phthalate esters in red and white wine samples.


"Double eyes" sign of congenital bilateral dacryocystoceles.
Dacryocystocele is caused by nasolacrimal duct obstruction and results in cystic dilatation of the proximal part of the nasolacrimal duct, which is located inferomedial to the orbit, leading to fluid accumulation. It is important to consider that persistent congenital bilateral dacryocystoceles may cause neonatal nasal obstruction resulting in respiratory difficulty, and large dacryocystoceles may require surgical drainage. Ultrasonography demonstrates that congenital bilateral dacryocystoceles and normal eyeballs prenatally resemble two pairs of cystic "lesions" of different sizes. We herein present a case of prenatally diagnosed isolated congenital bilateral dacryocystoceles and propose the new name of "double eyes" sign for this rare condition to create an impact on medical students and residents.


Studies on structural and immunogenic polypeptides of hydropericardium syndrome virus by SDS-PAGE and western blotting.
The polypeptide pattern of a local isolate of a virus causing hydropericardium syndrome was analyzed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. A total of 12 polypeptides ranging in molecular weight between 13.8 and 110.0 kDa were observed. Western blot analysis of structural polypeptides revealed seven immunogenic polypeptides ranging in molecular weight between 15.8 and 110.0 kDa.


Identification of Trypanosoma theileri as a contaminant in primary cultures of bovine retina.
T. theileri has been isolated from primary cultures of bovine retina and subcultered successfully for 2 passages in sub-confluent cultures. When cultures reached confluency no trypomastigotes or epimastigotes could be detected and attempts to recover trypanosomes from these cultures were unsuccessful. The presence of intracellular forms could not formally be excluded.


Gene expression profiling of cultured cells from brainstem of newborn spontaneously hypertensive and Wistar Kyoto rats.
The spontaneously hypertensive rat (SHR) is a good model to study several diseases such as the attention-deficit hyperactivity disorder, cardiopulmonary impairment, nephropathy, as well as hypertension, which is a multifactor disease that possibly involves alterations in gene expression in hypertensive relative to normotensive subjects. In this study, we used high-density oligoarrays to compare gene expression profiles in cultured neurons and glia from brainstem of newborn normotensive Wistar Kyoto (WKY) and SHR rats. We found 376 genes differentially expressed between SHR and WKY brainstem cells that preferentially map to 17 metabolic/signaling pathways. Some of the pathways and regulated genes identified herein are obviously related to cardiovascular regulation; in addition there are several genes differentially expressed in SHR not yet associated to hypertension, which may be attributed to other differences between SHR and WKY strains. This constitute a rich resource for the identification and characterization of novel genes associated to phenotypic differences observed in SHR relative to WKY, including hypertension. In conclusion, this study describes for the first time the gene profiling pattern of brainstem cells from SHR and WKY rats, which opens up new possibilities and strategies of investigation and possible therapeutics to hypertension, as well as for the understanding of the brain contribution to phenotypic differences between SHR and WKY rats.


Effect of Neurodegenerative Mutations in the NEFL Gene on Thermal Denaturation of the Neurofilament Light Chain Protein.
Effects of E90K, N98S, and A149V mutations in the light chain of neurofilaments (NFL) on the structure and thermal denaturation of the NFL molecule were investigated. By using circular dichroism spectroscopy, it was shown that these mutations did not lead to the changes in α-helical structure of NFL, but they caused noticeable effects on the stability of the molecule. We also identified calorimetric domains in the NFL structure by using differential scanning calorimetry. It was shown that the E90K replacement leads to the disappearance of the low-temperature thermal transition (domain 1). The mutations cause changes in the enthalpy of NFL domains melting, as well as lead to the significant changes in the melting temperatures (Tm) of some calorimetric domains. Thus, despite the fact that all these mutations are associated with the development of Charcot-Marie-Tooth neuropathy, and two of them are even located very close to each other in the coil 1A, they affect differently structure and stability of the NFL molecule.


Implementation of a combined bioinformatics and experimental approach to address lncRNA mechanism of action: The example of NRIR.
In this study, we demonstrate the benefit of applying combined strategies to analyze lncRNA action based on bioinformatics and experimental information. This strategy was developed to identify the molecular function of negative regulator of interferon response (NRIR), a type I interferon-stimulated gene (ISG), that we have previously demonstrated to be involved in the upregulation of a subset of ISGs in LPS-stimulated human monocytes. In this study, we provide experimental evidence that NRIR is localized in cellular nuclei, enriched on the chromatin fraction, and upregulates ISGs acting at the transcriptional level. In silico analysis of secondary structures identified distinct NRIR structural domains, comprising putative DNA- and protein-binding regions. In parallel, the presence of a putative DNA-binding domain in NRIR and the five putative NRIR-binding sites in the promoter of NRIR-target genes support the function of NRIR as a transcriptional regulator of its target genes. By use of integrated experimental/bioinformatics approaches, comprising database and literature mining together with in silico analysis of putative NRIR-binding proteins, we identified a list of eight transcription factors (TFs) shared by the majority of NRIR-target genes and simultaneously able to bind TF binding sites enriched in the NRIR-target gene promoters. Among these TFs, the predicted NRIR:STAT interactions were experimentally validated by RIP assay.


Prenatal diagnosis of cloacal anomalies.
To evaluate prenatal diagnosis in facilitating prenatal counselling and planning optimal perinatal care for persistent cloaca, a complex malformation with variable presentation and a difficult reconstructive challenge for the paediatric urologist and surgeon. The prenatal records of six patients with a suspected prenatal diagnosis of cloacal anomaly, subsequently confirmed on delivery, were reviewed. All had serial prenatal ultrasonograms. Fetal medicine and paediatric surgical specialists were present for the scans and counselled the parents jointly. The diagnosis was made at 19-33 weeks of gestation; all fetuses had a cystic structure arising from the pelvis and bilateral hydronephrosis, with a poorly visualized fetal bladder in most. Other prenatal features included transient fetal ascites, oligohydramnios, ambiguous genitalia and growth retardation. Female karyotype was confirmed in all cases by amniocentesis. Diagnosis was aided in two patients by fetoscopy and in another by magnetic resonance imaging. Cloacal anomalies can be diagnosed prenatally and should be considered in any female fetus presenting with bilateral hydronephrosis, a poorly visualized bladder and a cystic lesion arising from the pelvis. Prenatal diagnosis allows time for parental counselling and planning of the delivery at a centre equipped with neonatal intensive-care and paediatric surgical facilities.


Nerve injury affects the capillary supply in rat slow and fast muscles differently.
The goal of this study was to determine the acute effects of permanent denervation on the length density of the capillary network in rat slow soleus (SOL) and fast extensor digitorum longus (EDL) muscles and the effect of short-lasting reinnervation in slow muscle only. Denervation was performed by cutting the sciatic nerve. Both muscles were excised 2 weeks later. Reinnervation was studied 4 weeks after nerve crush in SOL muscle only. Capillaries and muscle fibres were visualised by triple immunofluorescent staining with antibodies against CD31 and laminin and with fluorescein-labelled Griffonia (Bandeira) simplicifolia lectin. A recently developed stereological approach allowing the estimation of the length of capillaries adjacent to each individual fibre (Lcap/Lfib) was employed. Three-dimensional virtual test grids were applied to stacks of optical images captured with a confocal microscope and their intersections with capillaries and muscle fibres were counted. Interrelationships among capillaries and muscle fibres were demonstrated with maximum intensity projection of the acquired stacks of optical images. The course of capillaries in EDL seemed to be parallel to the fibre axes, whereas in SOL, their preferential direction deviated from the fibre axes and formed more cross-connections among neighbouring capillaries. Lcap/Lfib was clearly reduced in denervated SOL but remained unchanged in EDL, although the muscle fibres significantly atrophied in both muscle types. When soleus muscle was reinnervated, capillary length per unit fibre length was completely restored. The physiological background for the different responses of the capillary network in slow and fast muscle is discussed.


Ssu72 phosphatase-dependent erasure of phospho-Ser7 marks on the RNA polymerase II C-terminal domain is essential for viability and transcription termination.
The C-terminal domain (CTD) of the largest subunit of RNA polymerase II (Pol II) serves an important role in coordinating stage-specific recruitment and release of cellular machines during transcription. Dynamic placement and removal of phosphorylation marks on different residues of a repeating heptapeptide (YSPTSPS) of the CTD underlies the engagement of relevant cellular machinery. Whereas sequential placement of phosphorylation marks is well explored, genome-wide engagement of phosphatases that remove these CTD marks is poorly understood. In particular, identifying the enzyme that erases phospho-Ser7 (Ser7-P) marks is especially important, because we find that substituting this residue with a glutamate, a phospho-mimic, is lethal. Our observations implicate Ssu72 as a Ser7-P phosphatase. We report that removal of all phospho-CTD marks during transcription termination is mechanistically coupled. An inability to remove these marks prevents Pol II from terminating efficiently and will likely impede subsequent assembly into the pre-initiation complex.


Methods and early recruitment of the Brain Attack Surveillance in Corpus Christi-Post-acute Care (BASIC-PAC) Project.
Compared with non-Hispanic whites (NHWs), Mexican Americans (MAs) have worse stroke outcomes. We report here the methods, background literature, and initial recruitment of the Brain Attack Surveillance in Corpus Christi-Post Acute Care (BASIC-PAC) Project which aims to explore PAC in MAs and NHWs from multiple perspectives: patients, caregivers, and community. Rigorous active and passive stroke surveillance captures all strokes in Nueces County, Texas. Stroke patients are followed for 90 days to determine their care transitions and factors influencing their rehabilitation setting. Informal caregivers of the stroke patients are identified and interviewed at 90 days to determine aspects of their caregiving and caregiver outcomes. Available community resources are compared with stated needs among stroke patient and caregivers to determine unmet needs. Between October, 2019 and October, 2021, among the 629 stroke patients eligible, 413 were MA, 227 were NHW. Of the 629, all of the six follow-up calls were completed by 355 of the MAs (87%) and 191 of the NHWs (87%). During this same time period, we attempted to approach 621 potential caregivers. Of these, 458 (73.8%) potential caregivers participated in interviews to determine caregiver eligibility, and 373 (81.4%) of these participating potential caregivers met the eligibility criteria. BASIC-PAC has strong initial recruitment and is poised to provide valuable data on multiple aspects of PAC and how PAC differs by ethnicity and contributes to worse stroke outcomes in MAs. Based on the study findings, interventions can be developed that will improve stroke health equity.


Ultrasound-assisted ionic liquid dispersive liquid-liquid microextraction coupled with high performance liquid chromatography for sensitive determination of trace celastrol in urine.
Ultrasound-assisted ionic liquid dispersive liquid-liquid microextraction (UA IL-DLLME) coupled with high-performance liquid chromatography (HPLC) has been developed for the determination of celastrol in human urine samples. In the microextraction procedure, ionic liquid (IL) was used as extraction solvent and dispersed into the aqueous sample solution as fine droplets by means of dispersive solvent and ultrasonication which promoted the analyte to migrate into IL phase more easily. Several important parameters affecting the extraction efficiency were studied and optimized, including the type and volume of extraction solvent and dispersive solvent, sample pH, ultrasonication time, cooling time, centrifugation time and salting-out effect. Under the optimized conditions, 110-fold enrichment factor was obtained and the limit of detection (LOD) was 1.6 μg/L at a signal-to-noise ratio of 3. The calibration curve was linear over the range of 10-1000 μg/L for celastrol in human urine sample, with a correlation coefficient of 0.9980. Intra- and inter-assay precision were 0.43% and 2.78%, respectively. The proposed method was successfully applied to the real human urine samples and good spiked recoveries in the range of 93.2-109.3% were obtained.


Ultrasound findings of subcutaneous soft tissue metastasis of renal cell carcinoma: A case report.
In renal cell carcinoma (RCC), metastasis to subcutaneous soft tissues is rare, with only a small number of cases reported to date, especially those based on ultrasound findings. This case report presents the ultrasound findings of an 81-year-old Chinese man who developed RCC presenting as subcutaneous soft-tissue metastasis 15 years after curative nephrectomy. We suggest that ultrasound-guided puncture biopsy should be considered when such suspicious lesions are found.


Making choices for childbirth: development and testing of a decision-aid for women who have experienced previous caesarean.
This paper presents the development and pilot testing of an evidence-based decision-aid for pregnant women who have experienced previous caesarean section and who are considering options for birth in a subsequent pregnancy. The Ottawa Decision Support Framework (DSF) has been utilised and modified in the development of a tailored booklet entitled "Birth Choices: What is best for you...vaginal or caesarean birth?" Development included a review by women who had experienced previous caesarean birth as well as obstetric, midwifery and educational experts. The booklet was also reviewed for acceptability by 21 pregnant women who were facing this birth decision. Pre- and post-booklet preferences for birth were elicited and compared to actual birth outcomes. This decision-aid shows promise in facilitating informed decision-making in pregnancy. It is currently undergoing an evaluation in a multi-centre randomised controlled trial.


Photo-CIDNP experiments with an optimized presaturation pulse train, gated continuous illumination, and a background-nulling pulse grid.
Methods to record chemically induced dynamic nuclear polarization (CIDNP) spectra that are virtually free from background magnetization and avoid the sensitivity loss and subtraction artifacts of difference spectroscopy have been developed. Presaturation by a string of composite pi/2 pulses, each followed by a defocussing field gradient, is analyzed, and guidelines for the optimization of pulse phases and gradient strengths are derived. Subsequent gated illumination during a grid of pi pulses with a prescribed timing causes the background magnetization to vanish at those moments of a pulse sequence when CIDNP magnetization is to be sampled or transferred. By shifting the illumination intervals within such a grid, the sign of the polarizations can be inverted without influencing the development of the background magnetization, allowing a further strong suppression of residual background by a phase cycle. Experimental examples for the application of these methods to more complex CIDNP experiments (1D-CIDNP-COSY, 1D-CIDNP-TOCSY, CIDNP-induced heteronuclear Overhauser effects, water suppression in protein CIDNP) are given.


Proteinases released by the parasitic larval stages of Ascaris suum, and their inhibition by antibody.
The tissue-invasive infective and lung-stage larvae of the nematode Ascaris suum were found to release proteinases during culture in vitro. This activity contained multiple proteolytic enzyme activities, as defined by pH optima, substrate specificities, and inhibitor profiles. Chymotryptic, tryptic collagenolytic and elastolytic activities were produced by both developmental stages, with major pH optima at pH 6 and 9, and there were indications of unusual interactions between the enzymes. The set of proteinases released was found to be specific to each stage of the parasite, although these included some activities which were indistinguishable between the products of the two. The in vitro-released materials of the tissue-parasitic stages of Ascaris are already known to be potently antigenic. Here, we found that this antigenicity was reflected by inhibition of the proteinases of both stages by serum antibody from infected animals. This inactivation of major secreted enzymes of this parasite could presumably contribute to impairment of survival and migratory potential in sensitised hosts.


Left ventricular remodeling and its correlates among adolescents with perinatally acquired HIV in South Africa.
Left ventricular (LV) remodeling and its transitions from compensatory adaptations to LV dysfunction have not been examined in adolescents with perinatally acquired HIV infection (PHIV). We used cardiovascular magnetic resonance (CMR) in a cross-sectional study to characterize PHIV-related progressive LV remodeling in adolescents in South Africa. Adolescents with PHIV on antiretroviral treatment and their HIV uninfected peers completed 3 T CMR examination. We defined LV remodeling by LV mass/volume (M/V) ratio, modelling progressive LV remodeling as increasing M/V ratio. Linear regression models were applied to estimate the correlates of progressive LV remodeling. Overall, 71 adolescents with PHIV [mean age: 15.2 years; 54% male] and 36 HIV uninfected [15.1 years; 42% male] peers were enrolled. Adolescents with PHIV had lower mean LV M/V ratio (0.68 vs. 0.75 g/mL; p = 0.004) than HIV uninfected peers, without LV hypertrophy in either group. Among adolescents with PHIV, increasing M/V ratio was accompanied by increasing interstitial volume [adjusted mean change (AMC) per 0.1 g/mL M/V ratio: 1.75 mL, p < 0.001] with no change in global circumferential strain (GCS) [AMC per 0.1 g/mL M/V ratio: -0.21%, p = 0.48]. However, in HIV uninfected individuals, increasing M/V ratio was accompanied by increasing peak GCS [AMC per 0.1 g/mL M/V ratio: -1.25%, p = 0.039] with no change in interstitial volume (AMC per 0.1 g/mL M/V ratio: 1.16 mL, p = 0.32]. Successfully treated PHIV is associated with less severe LV remodeling in adolescence when compared to HIV uninfected controls. LV remodeling in PHIV is associated with disproportionate expansion of the non-contractile interstitium not accompanied by improved GCS.


Gap junctions between lateral spinal motoneurons in the rat.
Ultrastructural examination reveals gap junctional plaques between motoneurons in the dorsolateral nucleus (DLN), an androgen-sensitive motor nucleus in the lumbar spinal cord of rats. This nucleus contains motoneurons innervating the ischiocavernosus muscle and urethral sphincter. Gap junctional plaques were found along the somatic and proximal dendritic membranes of DLN motoneurons, and between membranes of bundled dendrites in the neuropil of this nucleus. Castration and androgen treatment had no significant effect on the size or frequency of gap junctions.


Complementary Proteomic and Biochemical Analysis of Peptidases in Lobster Gastric Juice Uncovers the Functional Role of Individual Enzymes in Food Digestion.
Crustaceans are a diverse group, distributed in widely variable environmental conditions for which they show an equally extensive range of biochemical adaptations. Some digestive enzymes have been studied by purification/characterization approaches. However, global analysis is crucial to understand how digestive enzymes interplay. Here, we present the first proteomic analysis of the digestive fluid from a crustacean (Homarus americanus) and identify glycosidases and peptidases as the most abundant classes of hydrolytic enzymes. The digestion pathway of complex carbohydrates was predicted by comparing the lobster enzymes to similar enzymes from other crustaceans. A novel and unbiased substrate profiling approach was used to uncover the global proteolytic specificity of gastric juice and determine the contribution of cysteine and aspartic acid peptidases. These enzymes were separated by gel electrophoresis and their individual substrate specificities uncovered from the resulting gel bands. This new technique is called zymoMSP. Each cysteine peptidase cleaves a set of unique peptide bonds and the S2 pocket determines their substrate specificity. Finally, affinity chromatography was used to enrich for a digestive cathepsin D1 to compare its substrate specificity and cold-adapted enzymatic properties to mammalian enzymes. We conclude that the H. americanus digestive peptidases may have useful therapeutic applications, due to their cold-adaptation properties and ability to hydrolyze collagen.


Flow karyotypes and chromosomal DNA contents of genus Triticum species and rye (Secale cereale).
The flow cytometry and chromosome imaging method were jointly used for analyzing genome content and chromosomal DNA content of hexaploid wheat (AABBDD), hexaploid triticale (AABBRR), tetraploid wheat (AABB), and AA, BB, DD genome donors and RR genome rye. Their genome sizes were 34.4 pg, 40.9 pg, 26.2 pg, 12.1 pg, 13.7 pg, 10.5 pg, and 16.9 pg, respectively. The 2C nuclear DNA content of BB genome donor with 13.7 pg was the highest value among the other genome donors, AA or DD. The genome content of tetraploid wheat, unlike hexaploid wheat or hexaploid triticale, was larger than the sum of the genomes of AA and BB genome donors. The DNA content of each chromosome ranged from 1.22 pg in DD genome donor to 2.61 pg in rye. Each chromosome peak was divided into three to four groups. Only one chromosome was included in the highest chromosomal DNA peak in hexaploid wheat, tetraploid wheat, DD genome donor and rye but two chromosomes in AA, BB genome donors, and hexaploid triticale. Correlation between 2C nuclear DNA content and chromosome density volume was the highest value compared with the other chromosomal parameters of chromosome area, or chromosome length.


Hormone-sensitive particulate cAMP phosphodiesterase activity in 3T3-L1 adipocytes. Regulation of responsiveness by dexamethasone.
Confluent 3T3-L1 fibroblasts incubated for 72 h with methylisobutylxanthine, dexamethasone, and insulin differentiate and acquire phenotypic characteristics of mature adipocytes, including hormone-sensitive cAMP phosphodiesterase activity located in a particulate fraction of homogenates. About 10 days after initiating differentiation, a maximally effective concentration of insulin (100 pM) increased particulate cAMP phosphodiesterase activity 40 to 60% in 8 min; activation persisted for at least 30 min in the presence of insulin. Incubation of adipocytes for 6-8 min with agents that increased cAMP, e.g. 1 microM epinephrine, 0.1 microM isoproterenol, corticotropin (2 mu units/ml), or thyroid-stimulating hormone (15 ng/ml), also increased particulate phosphodiesterase activity 40-60%. Changes in phosphodiesterase activity produced by epinephrine tended to lag behind changes in cAMP. Insulin, epinephrine, and corticotropin increased Vmax, not Km (0.5 microM), for cAMP. Particulate phosphodiesterase activity, solubilized with detergent, eluted in a single peak from DEAE-Bio-Gel. Insulin and epinephrine increased the activity eluted in this peak. Neither insulin nor lipolytic hormones increased activity in soluble fractions from differentiated cells or particulate or soluble fractions from undifferentiated cells. Incubation of adipocytes for 48 h with 1 microM dexamethasone prevented insulin-induced activation of the particulate phosphodiesterase and did not alter basal activity. After incubation for 72 h with 0.1 microM dexamethasone, insulin and epinephrine activation were abolished. These effects of dexamethasone on hormonal regulation of particulate phosphodiesterase activity could account for some of the so-called permissive effects of glucocorticoids on cAMP-mediated processes as well as the "anti-insulin" effects of glucocorticoids.


Atypical Cause of Sepsis from Bilateral Iliopsoas Abscesses Seeded from Self-mutilation: A Case Report.
An iliopsoas abscess (IPA) is an abscess located adjacent to the iliopsoas and iliacus muscles. Although rare, their variable clinical presentations often lead to a delay in diagnosis. We present a case of sepsis secondary to multiple IPAs that was missed despite multiple healthcare encounters. The patient had no classical risk factors for an IPA, and the abscesses were found to be seeded via hematogenous spread from self-inflicted cutting. This case illustrates the importance of obtaining a complete history, including psychiatric screen, and performing a thorough examination when evaluating patients with low back pain to rule out overlooked sources of bacteremia.


Long-term (>10-year) clinical follow-up after young embolic stroke/TIA of undetermined source.
To date, the pathophysiology of first-ever and recurrent stroke/TIA still remains unclear in young patients with embolic stroke/TIA of undetermined source (ESUS). Clinical studies with long-term follow-up in young ESUS patients are necessary to investigate the underlying pathophysiology of first-ever and recurrent stroke/TIA in this patient population, in particular the role of new-onset atrial fibrillation. Our aim was to study the long-term (>10-year) clinical outcome of young patients (<50 years) with ESUS. This cohort study included all patients aged ≤ 50 years who underwent transoesophageal echocardiography for diagnostic work-up of ESUS during 1996-2008 from one tertiary center. All patients were contacted by telephone between September-November 2018 to update clinical information from medical records. The clinical outcomes of this study were incidence rates of all-cause and cardiovascular mortality, recurrent stroke/TIA, new-onset clinical AF, and ischemic vascular events. In total, 108 patients (57% female, mean age 40 ± 7.2 years [range 19-50 years], n = 72 stroke) were included. Across clinical follow-up (median 13[IQR 10-16] years), 24 patients died (n = 14 cardiovascular). The 15-year incidence rate of recurrent stroke/TIA was 15% (incidence rate = 1.09[95%CI 0.54-1.65]/100 patient-years) and a 5.5% incidence of new-onset clinical AF (incidence rate = 0.44[95%CI 0.09-0.79]/100 patient-years) following ESUS. The incidence of recurrent stroke/TIA is relatively high during long-term clinical follow-up of young patients with ESUS. In contrast, new-onset clinical AF is relatively low and therefore may not play an important part in the pathophysiology of first-ever and recurrent stroke/TIA of these patients.


Macular pigment and the colour-specificity of visual evoked potentials.
A modified Moreland anomaloscope was used to examine two subjects, one with dense macular pigment, the other with relatively light pigmentation. Chromatic visual evoked potentials (VEPs) were elicited from these two subjects using coarse, isoluminant gratings of different sizes. Colour-specificity was verified by comparing chromatic onset VEPs (reflecting sustained activity) with chromatic reversal VEPs (reflecting mainly transient activity) and also by Fourier analysis (colour-specific responses are dominated by the fundamental, transient activity by the second harmonic). Achromatic (transient-type) intrusions, produced by large blue-green gratings could be related to the extent of subject-specific macular pigmentation.


Metachronous bilateral ovarian teratoma: a germ-line familial disorder and review of surgical management options.
Germ cell tumors in females are uncommon, and bilateral metachronous ovarian teratoma is even exceptional, with sporadic cases described in the literature. We report on a girl in whom a metachronous ovarian teratoma occurred 6 years after the first. The simultaneous onset of germ-line anomalies in other members of the family supports the existence of genetic or environmental factors conferring susceptibility to germ cell lesions. The case here illustrated reminds the issue of the appropriate follow-up of these patients and of their families.


Involuntary Celibacy: A Review of Incel Ideology and Experiences with Dating, Rejection, and Associated Mental Health and Emotional Sequelae.
Incels (involuntary celibates) have recently garnered media attention for seemingly random attacks of violence. Much attention has centered around the misogynistic and violent discourse that has taken place in online incel forums as well as manifestos written by incels who have perpetrated deadly attacks. Such work overlooks the experiences and issues faced by incels themselves, the majority of which have not engaged in any violent behavior. A small number of studies have recruited incels. Results from these studies highlight the nuanced nature of the incel identity. It is also apparent that incels suffer from high levels of romantic rejection and a greater degree of depressive and anxious symptoms, insecure attachment, fear of being single, and loneliness. Incels report significant issues pertaining to their mental, social, and relational well-being and may seek support from forums that often feature misogynistic and violent content.


The effects of release and depletion of endogenous noradrenaline on the transmission of impulses in the mouse vas deferens.
1 The effects of endogenous noradrenaline released by tyramine and the influence of depletion of the tissue noradrenaline with reserpine and/or alpha-methyl-p-tyrosine on the twitch responses of the field-stimulated mouse vas deferens have been studied.2 Tyramine (10-40 muM) inhibited the twitch responses to field stimulation and failed to produce a contraction. The inhibition decreased as the rate of stimulation increased.3 The inhibition produced by tyramine was antagonized by cocaine (10 muM) and by yohimbine (10 nM), which indicated that it was produced by released noradrenaline acting on presynaptic alpha-adrenoceptors.4 Depletion of the tissue noradrenaline by 39% by blockade of the synthesis of noradrenaline with alpha-methyl-p-tyrosine, was without effect on the twitch response but it reduced the inhibitory effect of tyramine.5 Depletion of the tissue noradrenaline by 96.5% with reserpine alone and by 99.4%, with a combination of reserpine and alpha-methyl-p-tyrosine, reduced the twitch responses by approximately 66% and virtually abolished the inhibition produced by tyramine. It also increased the rate of decline of the responses when the tissue was continuously stimulated. The remaining twitch was not antagonized by phenoxybenzamine (15 muM).6 Residual twitches were bigger in tissues depleted by 99.4% than in those depleted by only 96.5%. This difference was eliminated in the presence of yohimbine (128 nM).7 It is concluded that inhibition of the twitch responses by tyramine is produced by stimulation of presynaptic alpha-adrenoceptors and that the twitch response is associated with stimulation of the sympathetic neurone, but that it is not mediated by postsynaptic alpha-adrenoceptors.


The Contribution of Gestational Diabetes to Diabetes Risk Among First Nations and Non-First Nations Women in Saskatchewan: Results From the DIP: ORRIIGENSS Project.
Our aim in this study was to determine the risk for diabetes mellitus (DM) among Saskatchewan First Nations (FN) and non-FN women with prior gestational DM (GDM). Using Ministry of Health administrative databases, we conducted a retrospective cohort study of DM risk by GDM occurrence among FN and non-FN women giving birth from 1980 to 2009 and followed to March 31, 2013. We determined frequencies and odds ratios (ORs) of DM in women with/without prior GDM after stratifying by FN status, while adjusting for other DM determinants. Survival curves of women until DM diagnosis were obtained by prior GDM occurrence and stratified by ethnicity and total parity. De-identified data were obtained for 202,588 women. Of those who developed DM, 2,074 of 10,114 (20.5%) had previously experienced GDM (811 of 3,128 [25.9%]) FN and 1,263 of 6,986 [18.1%] non-FN). Cumulative survival of women with prior GDM until DM was higher for FN than for non-FN women (82% vs 46%), but prior GDM was a stronger predictor of DM within the non-FN cohort (prior GDM vs no GDM: OR, 9.64 for non-FN; OR, 7.05 for FN). Finally, higher total parity interacted with prior GDM to increase DM risk in both groups. With prior GDM and parity ≥3, 93% of FN and 57% of non-FN women subsequently developed DM. GDM is a leading determinant of T2DM among FN and non-FN women, amplified by higher parity. This contributes to earlier onset diabetes, affecting subsequent pregnancies and increasing risk for chronic diabetic complications. It may also factor into higher type 2 DM rates observed in FN women compared with men.


Lack of topoisomerase copy number changes in patients with de novo and relapsed diffuse large B-cell lymphoma.
Topoisomerase (TOP) gene copy number changes may predict response to treatment with TOP-targeting drugs in cancer treatment. This was first described in patients with breast cancer and is currently being investigated in other malignant diseases. TOP-targeting drugs may induce TOP gene copy number changes at relapse, with possible implications for relapse therapy efficacy. TOP gene alterations in lymphoma are poorly investigated. In this study, TOP1 and TOP2A gene alterations were investigated in patients with de novo diffuse large B-cell lymphoma (DLBCL) (n = 33) and relapsed DLBCL treated with chemotherapy regimens including TOP2-targeting drugs (n = 16). No TOP1 or TOP2A copy number changes were found. Polysomy of chromosomes 20 and 17 was seen in 3 of 25 patients (12%) and 2 of 32 patients (6%) with de novo DLBCL. Among relapsed patients, chromosome polysomy was more frequently observed in 5 of 13 patients (38%) and 4 of 16 patients (25%) harboring chromosome 20 and 17 polysomy, respectively; however, these differences only tended to be significant (p = 0.09 and p = 0.09, respectively). The results suggest that TOP gene copy number changes are very infrequent in DLBCL and not likely induced by TOP2-targeting drugs. Increased polyploidy of chromosomes 17 and 20 among patients with relapsed DLBCL may reflect genetic compensation in the tumor cells after TOP2 inhibition, but is more likely due to the increased genetic instability often seen in progressed cancers. Therefore, it is unlikely that TOP1 and TOP2A gene alterations can be used as predictive markers for response to treatment with TOP2-targeting drugs in patients with DLBCL.


Paradoxical reaction to antituberculosis therapy after 6 months of treatment for pulmonary tuberculosis: A case report.
Paradoxical reactions (PRs) to antituberculosis (anti-TB) drugs during treatment are well known phenomena, but a PR presenting as a new pulmonary lesion after completion of treatment is extremely rare, and little is known about the management of such cases. A 44-year-old man was diagnosed with pulmonary TB. His sputum cultures became negative 45 days after the initiation of standard anti-TB treatment. Upon the patient's completion of 6 months of anti-TB therapy, computed tomography revealed a new irregularly shaped mass in the lower left pulmonary lobe. A transbronchial lung biopsy (TBLB) revealed caseous necrosis and granulomatosis surrounded by epithelioid and multinucleated giant cells. Cultures of both the TBLB specimen and bronchoalveolar lavage fluid remained negative for TB. The CT shadow disappeared 6 months later without further administration of anti-TB drugs. Careful observation without therapy may be sufficient for a patient treated for TB who develops a PR upon completion of treatment, if the patient has achieved a bacteriological remission.


A novel biopesticide PONNEEM to control human vector mosquitoes Anopheles stephensi L. and Culex quinquefasciatus Say.
Organophosphate pesticides are widely used in vector mosquito management and agricultural pest management. These chemicals enter into natural water bodies and soil and cause hazards to the environment. The objective of this study was to prepare a natural pesticide which will not harm the environment and yet control vector mosquitoes. PONNEEM, a novel biopesticide, patented and prepared from the oils of Azadirachta indica and Pongamia glabra, was tested against Anopheles stephensi and Culex quinquefasciatus. One hundred percent larvicidal and ovicidal activities were observed at 0.1-ppm concentration of PONNEEM against the two mosquito species under laboratory and sunlight conditions up to 12 months from the date of manufacture. Very high oviposition reduction of 26.46 and 32.16 % is also recorded. Reductions in α-esterase level (0.0818 ± 0.340 and 0.2188 ± 0.003), β-esterase level (0.0866 ± 0.026 and 0.0398 ± 0.010 μg naphthol produced/min/mg larval protein), glutathione S-transferase enzyme (14.2571 ± 0.51 and 15.3326 ± 0.51 μmol/min/mg larval protein) and total protein levels (0.0390 ± 0.008 and 0.1975 ± 0.029 mg/individual larva in treated groups of A. stephensi and C. quinquefasciatus at 0.1-ppm concentration, respectively. The non-target organisms such as Gambusia affinis and Diplonychus indicus were not affected. Biopesticides are good alternatives to synthetic pesticides. PONNEEM can be effectively used for the management of human vector mosquitoes. Since it has a biodegradable nature and does not alter the environmental condition of water and soil.


Short-term effects of the Sander bite-jumping appliance on the pharyngeal airways in subjects with skeletal Class II malocclusion: A retrospective case-control study.
Skeletal Class II subjects present often a retruded mandible that might increase the probability of breathing disorders. To evaluate the effects of functional treatment by means of the Sander bite-jumping appliance (BJA) on the upper airways of growing subjects. Thirty-four subjects (21 males, 13 females; mean age 11.1 ± 1.2 years) with skeletal Class II malocclusion due to mandibular retrusion were treated with the Sander BJA (BJA group). The control group consisted of thirty-four untreated subjects with skeletal Class II malocclusion (25 males, 9 females; 10.4 ± 1.2 years; CTR group). The effect of the Sander BJA appliance on the airway dimensions was evaluated by comparing lateral cephalograms recorded before (T0) and after (T1) the period of observation/treatment. Within- and between-group comparisons were statistically evaluated according to the distribution of the data (P < .05). The oropharyngeal region (ph_pph) increased in the BJA group (1.8 ± 3.2; P = .001) but the difference respect to the CTR group was not statistically significant (0.73 ± 4.0; P = .314; P = .077). The tongue height (th) increased in the BJA group (2.8 ± 3.7; P < .001), and this increase was statistically significant respect to the CTR group (0.6 ± 4.7; P = .461; P = .038). In the BJA, the Ans_Pns_P angle showed a statistically significant decrease (-3.8 ± 7.8; P = .007) compared to the CTR group (-0.1 ± 5.4; P = .705; P = .026). The airway dimensions increased for both control subjects and Class II patients treated with Sander BJA due to physiological growth. The Sander BJA induced a statistically significant change in the tongue and soft palate position, but the clinically relevant of these changes is questionable.


Choosing Castration: A Thematic Analysis of the Perceived Pros and Cons of Genital Injuries and Ablation by Men Who Voluntarily Sought Castration.
Some men elect castration voluntarily without any clear medical reason. Here we aim to document their perception of genital ablation and injuries to better understand their motivations for castration. Participants completed an online survey with open-ended questions related to their perspectives on castration, genital ablation, and genital injuries. Thematic analyses were performed on the responses to these questions. Responses were obtained from 208 male castrated individuals (51.9 ± 16.0 years old). Among these, 154 were physically castrated, 36 chemically castrated, and 18 nullified (had testicles and penis removed). The majority learned about castration from media (55.8%) or animal castration (23.4%). The circumstances when they first wanted to be castrated varied greatly. Most (46.3%) wished to achieve an idealized self motivated by gender dysphoria, body integrity dysphoria, or wanting to be conspicuously non-sexual. The top themes we identified related to the respondents' perceptions of the pros of genital ablation were physical appearance, psychological benefit (i.e., a "eunuch calm"), and being non-sexual. Conversely, themes related to the cons they saw in having no genitals ranged from no disadvantages to loss of sexual/reproductive capability. Some perceived performing genital injury as a step toward ultimate castration or nullification. The respondents similarly varied in whether they saw any loss in having non-functional testicles. Perceptions in this regard appeared to differ depending on whether the respondents were taking supplemental androgens post-castration. Motivations for castration vary greatly between individuals. Clinicians need to understand men's diverse perceptions on castration in order to provide appropriate care for individuals with strong castration desire.


Silver development in microscopy and bioanalysis: a new versatile formulation for modern needs.
Potentially, silver development could unify most modern demands for clean, accurately localized marker amplification in microscopy and bioanalysis. However, the existing technology leaves room for improvement in developer design. A new formulation has been devised which, by using principles of silver chelation, avoids problems of self-nucleation and catalysis by light. It is made, just before use, by mixing together equal amounts of stock solutions containing high molarity, Tris-buffered silver nitrate and alcoholic, buffered pyrogallol. The two stocks are easily prepared and have very long shelf-lives. The developer is light insensitive for up to an hour at room temperature, so that development can proceed under ambient light conditions and at the neutral pH most suited to biological systems. The powerful reducer in the suggested formulation should allow the detection of low concentrations of marker signal in a wide range of applications.


Identifying the shared metabolic objectives of glycerol bioconversion in Klebsiella pneumoniae under different culture conditions.
This paper addresses the problem of identifying the shared metabolic objectives of glycerol bioconversion in Klebsiella pneumoniae for production of 1,3-propanediol (1,3-PD) under different culture conditions. To achieve this goal, we propose a multi-level programming model. This model includes three optimization problems, where the constraint region of the first level problem is implicitly determined by the other two optimization problems. The optimized objectives of the first and second level problems are to minimize the set of fluxes that are of major importance to glycerol metabolism and the difference between the observed fluxes and those computed by the model, respectively. The third level problem in the proposed multi-level programming simultaneously solves a set of flux balance analysis (FBA) models. A method is proposed to solve efficiently the presented multi-level programming problem. In this method, we first transform the proposed multi-level problem into a bi-level problem by applying the dual theory of linear programming to the FBA models of the third level. Next, the optimal solution of the above bi-level problem is obtained by iteratively solving a sequence of mixed integer programming problems. Optimization results reveal that the proposed method can identify the shared metabolic objectives of glycerol bioconversion in Klebsiella pneumoniae under three groups of experimental data.


Preliminary characterization of in vitro synthesized hypothalamic precursor ACTH/beta-endorphin-like material.
We have previously shown that cultured bovine pituitary or hypothalamic cells incorporate 3H-labeled amino acids into high molecular weight glycoproteins containing the antigenic determinants of both corticotropin (ACTH) and beta-endorphin. We now report resolution of this 3H-labeled ACTH/beta-endorphin-like material into two predominant size classes upon SDS polyacrylamide-gel electrophoresis with apparent molecular weights (Mr) of 41 500 +/- 1600 and 36 000 +/- 1100. Isoelectric focusing revealed these components to be basic proteins with apparent isoelectric points greater than 8.5. Overnight trypsinization generated a 3H-labeled fragment comigrating with synthetic beta-lipotropin (61-69) [beta-endorphin (1-9)] upon paper electrophoresis which was immunoprecipitable with antibody directed against the corresponding synthetic fragment. Limited trypsinization of bovine immunoreactive ACTH/beta-endorphin extracted from freshly obtained bovine hypothalamic and anterior pituitary tissue, generated fragments which possessed ACTH bioreactivity. Both bovine pituitary and hypothalamic derived material are similar with respect to these foregoing physiochemical parameters and appear to be larger than the reported forms in mouse pituitary.


Transfer in care and diabetes distress in young adults with type 1 diabetes mellitus.
Diabetes distress (DD) is a serious problem in many people with diabetes and is associated with unfavorable clinical and psychosocial outcomes in children and adults. Little is known about DD in young adults (YAs) with type 1 diabetes mellitus (T1DM) who transferred to adult care. This study aimed to explore the differences between YAs with/without DD regarding transfer experiences, self-management and health-related quality of life (HRQoL). Cross-sectional online questionnaire completed by YAs with T1DM after transfer. DD was measured with the short-form Problem Areas in Diabetes scale. Descriptive analyses were followed by t-tests and χ2 tests to explore differences between the groups with/without DD. Effect sizes were calculated. Of 164 respondents with mean age 22.7 (±1.56) years, 60.7% was female. The total sample scored low on DD (6.52±4.67; range: 0-17), but 57 (34.8%) had a score ≥8, indicating DD. YAs with DD felt less ready to transfer to adult care than those without DD and scored lower on alliance between pediatric and adult care and reception in adult care. They also reported poorer self-management skills and lower HRQoL in all domains of functioning. More than one-third YAs experienced DD after transfer; this was associated with less favorable transition, self-management and psychosocial outcomes. Transfer in care seems to be a source of DD. Systematic screening on DD and attention for YAs' worries is recommended in both pediatric and adult care.


Nematodirus battus: permeability changes, calcium binding, and phosphorylation of the eggshell during hatching.
Eggshells of Nematodirus battus leaked trehalose 4 hr after being stimulated to hatch, and became permeable to trypan blue at their poles; 80% of eggs were stained blue 24 hr later. Exogenous application of ruthenium red significantly inhibited chill- and sodium fluoride-stimulated hatching, 50% hatch inhibition occurring in 44.67 +/- 2.2 and 8.5 +/- 1.5 microM, respectively. Lanthanum chloride, however, was not as inhibitory as ruthenium red on fluoride-stimulated hatching, 50% occurring at 31.60 +/- 1.25 microM. A Scatchard plot of the competitive binding of ruthenium red to eggshells demonstrated a high-affinity binding site for calcium, KCa' = 1.92 microM and a second, low-affinity site, KCa" = 1169.60 microM. Ruthenium red binding was significantly reduced by several enzymes, e.g., EGTA-buffered trypsin reduced binding by 73%. Radioiodinated concanavalin A also bound competitively to the eggshells in the presence of alpha-D-glucosyl-alpha-D-glucopyranoside and alpha-methyl-D-mannopyranoside. Eggshells incorporated phosphorus-32 from ATP after chilling or on exposure to sodium fluoride; gel filtration of solubilized homogenates of these samples showed that two proteins were radiolabelled with molecular weights of 38 X 10(3) and 8 X 10(3) Da, respectively. This phosphorylation was inhibited by N-ethylmaleimide, which also prevented hatching.


Large hepatocellular carcinomas: treatment with transarterial chemoembolization alone or in combination with percutaneous cryoablation.
To evaluate the safety and efficacy of transarterial chemoembolization (TACE) combined with cryoablation (TACE-cryoablation) in large (main tumor ≥5 cm in diameter) hepatocellular carcinomas (HCCs). From January 2010 to December 2015, 56 lesions in 56 patients were treated with combination therapy via a single TACE session followed by one to three percutaneous cryoablation sessions twice a week (TACE-cryoablation group). A total of 54 lesions in 54 patients were treated with TACE alone for two to six sessions once a month (TACE group). The decision between TACE and TACE cryoablation was based on patient choice. Outcomes of patients in two groups were compared according to the largest tumor diameter (subgroup): Group A (5 cm ≤ tumor <10 cm), Group B (10 cm ≤ tumor <15 cm), and Group C (tumor ≥15 cm). The mean number of cryoablation sessions per patient was 2.3 (range: 1-6). Within Group B, TACE-cryoablation significantly improved survival compared with TACE alone (11.0 vs 6.0 months; p = .008). This was also seen in Group C (8.0 vs 5.0 months; p = .001). However, no significant difference was noted in Group A (17.0 vs 13.0 months; p = .674). The complications related to TACE were comparable between the two groups. Two adverse events of grade 3 - 4 related to cryoablation occurred in two patients (3.6%). The independent prognostic factors for survival included: TACE cryoablation, AFP level, main tumor size and extrahepatic metastasis. TACE-cryoablation may improve overall survival in patients with HCC who presented with a tumor diameter ≥10 cm, with minimal complications, when compared with TACE alone.


Wavelet subband discriminator for efficient unsupervised chest X-ray image restoration.
Chest X-ray (CXR) images are commonly used to show the internal structure of the human body without invasive intervention. The quality of CXR is an important factor as it affects the accuracy of a clinical diagnosis. Unfortunately, it is difficult to always get good quality CXR scans due to noises and scatters. Recently, wavelet directional CycleGAN (WavCycleGAN) has shown promising results in image restoration tasks by removing noise and artifacts without sacrificing high-frequency components of the input image. Unfortunately, WavCycleGAN directly reconstructs wavelet directional images that require a wavelet transform in both the training and test phases, resulting in additional processing steps and unnatural artifacts originating from the wavelet domain image. In addition, WavCycleGAN can only process artifact-related subbands, so it is difficult to apply WavCycleGAN when different levels of artifacts are present in all subbands. To address this, here we present a novel unsupervised CXR image restoration scheme with similar or even better artifact removal performance than WavCycleGAN in spite of wavelet transform being only applied in the training phase. We introduce a novel wavelet subband discriminator which can be combined with CycleGAN or switchable CycleGAN, where wavelet transform is applied only in the training phase for discriminators to match the distribution of wavelet subband components. In our framework, the image restoration network can be still applied in the image domain to prevent unnatural artifacts of the wavelet domain image with the help of the image-domain cycle-consistency loss. In addition, using wavelet subband discriminator makes it possible to remove artifacts in all subbands by utilizing frequency-specific wavelet subband discriminators. Through extensive experiments for noise and scatter removal in CXRs, we confirm that our method provides competitive performance compared to existing approaches without additional processing steps in the test phase. Furthermore, we show that our wavelet subband discriminator combined with the switchable CycleGAN can provide the flexibility by generating different levels of artifact removal. The proposed wavelet subband discriminator can be combined with the existing CycleGAN or switchable CycleGAN structures to construct an efficient unsupervised CXR image reconstruction. The advantage of our wavelet subband discriminator-based CXR image restoration is that, unlike traditional WavCycleGAN, it does not require any additional processing steps in the testing phase and does not generate unnatural artifacts originating from the wavelet domain image. We believe that our wavelet subband discriminator can be applied to various CXR image applications.


Ventral onlay buccal mucosa urethroplasty: a 10-year experience.
To report our experience, and to evaluate the long-term outcomes and complication profiles of ventral onlay buccal mucosal graft urethroplasty (BMU) after prior urological intervention. We retrospectively reviewed 114 consecutive patients between February 2001 and April 2009 who underwent buccal mucosal graft urethroplasty for recurrent anterior urethral stricture disease. Seven patients were excluded for incomplete data. The remaining 107 patients comprised the study cohort. The mean follow-up time was 39.3 months (range 6.6-127.3 months). All patients had prior urological attempts at operative management. The mean stricture length was 3.14 cm (range 1.0-8.0 cm). Indications for buccal mucosal graft urethroplasty included: lichen sclerosis (2.8%), iatrogenic (24.3%), infection (4.7%) and perineal trauma/straddle injury (20.6%). Of these patients, 78 had bulbo-membranous stricture disease, 20 had penile involvement and nine were multifocal strictures. The average number of prior urological procedures was 2.83 (range 1-9). The overall graft failure rate was 6.5%. Importantly, the re-operation rate was 20.6%, primarily for stricture recurrence (18), meatal stenosis (3) and urethral diverticulum. The mean time to complication was 10.8 months. Ventral onlay buccal mucosal graft urethroplasty offers satisfactory results in the setting of recurrent and complicated urethral stricture disease. Graft failures and complications generally occur within the first year after surgery. Bulbar strictures enjoy greater graft patency and lower complication rates than other stricture locations. In particular, guarded expectations should be given for stricture length >4 cm and multifocal disease.


Continuous operation of a fluidized bed reactor for the removal of estrogens by immobilized laccase on Eupergit supports.
The feasibility of the operation of a fluidized bed reactor for the removal of estrogens by immobilized laccase was investigated in order to improve the degradation yields and enzyme stability previously obtained with packed bed reactors. High removal levels (between 76 and 90%) and significantly prolonged stability of the biocatalyst over 16 days were attained. In parallel, a decrease up to 90% in the estrogenic activity of the effluent was measured. Thus, the technology presented seems a promising tool to increase the applicability of laccases in bioremediation processes.


Transgenic rescue of phenotypic deficits in a mouse model of alternating hemiplegia of childhood.
Missense mutations in ATP1A3 encoding Na(+),K(+)-ATPase α3 are the primary cause of alternating hemiplegia of childhood (AHC). Most ATP1A3 mutations in AHC lie within a cluster in or near transmembrane α-helix TM6, including I810N that is also found in the Myshkin mouse model of AHC. These mutations all substantially reduce Na(+),K(+)-ATPase α3 activity. Herein, we show that Myshkin mice carrying a wild-type Atp1a3 transgene that confers a 16 % increase in brain-specific total Na(+),K(+)-ATPase activity show significant phenotypic improvements compared with non-transgenic Myshkin mice. Interventions to increase the activity of wild-type Na(+),K(+)-ATPase α3 in AHC patients should be investigated further.


Ventricular defibrillation in canines with chronic infarction, and effects of lidocaine and procainamide.
Prior studies in dogs with normal hearts have demonstrated that lidocaine increases but procainamide does not change the energy required for successful defibrillation. Because many postinfarct patients receiving implantable cardioverter defibrillator devices require adjunctive antiarrhythmic therapy, we have studied the effects of lidocaine and procainamide on the relationship between delivered voltage and defibrillation success in mongrel dogs 21 +/- 3 days following ligation of the left anterior descending and first diagonal coronary arteries. Internal defibrillation testing using a patch-patch electrode configuration was performed before and during the administration of saline controls (n = 10), lidocaine (n = 10) and procainamide (n = 10). The mean infarct size as determined by staining with tetrazolium was 13.4% +/- 8.3% of right and left ventricles, and did not differ significantly between groups. The 50% effective defibrillation (ED50) voltage increased with infusions of saline (16% +/- 15%), lidocaine (40% +/- 22%), and procainamide (13% +/- 15%) and the ED50 energy increased 41% +/- 44%, 104% +/- 62%, and 35% +/- 36%, respectively. However, the increase in ED50 voltages and energies were significantly greater in animals receiving lidocaine compared to those receiving either saline control or procainamide (P < 0.01). There were trends toward change of hemodynamic parameters in all animals following baseline defibrillation testing; stroke volume declined 21% +/- 16%; and mean pulmonary artery and aortic pressure increased by 22% +/- 25% and 11% +/- 15%, respectively. In conclusion, unlike our previous studies in dogs with normal hearts, in this model hemodynamic deterioration occurred with repeated fibrillation and defibrillation, and defibrillation voltage requirements increased in the control series. Taking into consideration the increase in defibrillation voltage requirements over the duration of the experiments, lidocaine increases and procainamide does not change ED50; thus, their effects are similar in normal and infarcted canine hearts.


Antioxidant flavonoids from Epimedium wushanense.
Two new flavonoids, wushanicaritin (1) and wushankaempferol (2), along with 24 known flavonoids were isolated from the whole herb of Epimedium wushanense T.S. Ying (Berberidaceae). On the basis of NMR and ESI-MS spectroscopic analysis, structures of compounds 1 and 2 were elucidated as 8-γ-hydroxy-γ,γ-dimethylpropyl-3,5,7-trihydroxy-4'- methoxyflavone and kaempferol 3-O-α-l-[2,3-di-O-β-D-(6-E-p-coumaroyl) glucopyranosyl]-rhamnopyranosyl-7-O-α-L-rhamnopyranoside, respectively. DPPH radical scavenging activity tests indicated that 1 (IC(50) 35.3 μM) exhibited antioxidant activity comparable to Vitamin C (IC(50) 32.0 μM), while 2 (IC(50) 443.7 μM) showed weak activity.


Characterization of an efficient CRISPR-iCas9 system in Yarrowia lipolytica for the biosynthesis of carotenoids.
The application of clustered regularly interspaced short palindromic repeats-Cas (CRISPR-Cas9) technology in the genetic modification of Yarrowia lipolytica is challenged by low efficiency and low throughput. Here, a highly efficient CRISPR-iCas9 (with D147Y and P411T mutants) genetic manipulation tool was established for Y. lipolytica, which was further utilized to integrate carotene synthetic key genes and significantly improve the target product yield. First, CRISPR-iCas9 could shorten the time of genetic modification and enable the rapid knockout of nonsense suppressors. iCas9 can lead to more than 98% knockout efficiency for NHEJ-mediated repair after optimal target disruption of a single gene, 100% knockout efficiency for a single gene-guided version, and more than 80% knockout efficiency for multiple genes simultaneously in Y. lipolytica. Subsequently, this technology allowed for rapid one-step integration of large fragments (up to 9902-bp) of genes into chromosomes. Finally, YL-ABTG and YL-ABTG2Z were further constructed through CRISPR-iCas9 integration of key genes in a one-step process, resulting in a maximum β-carotene and zeaxanthin content of 3.12 mg/g and 2.33 mg/g dry cell weight, respectively. Therefore, CRISPR-iCas9 technology provides a feasible approach to genetic modification for efficient biosynthesis of biological compounds in Y. lipolytica. KEY POINTS: • iCas9 with D147Y and P411T mutants improved the CRISPR efficiency in Y. lipolytica. • CRISPR-iCas9 achieved efficient gene knockout and integration in Y. lipolytica. • CRISPR-iCas9 rapidly modified Y. lipolytica for carotenoid bioproduction.


The inhibition effect of natural food supplement active ingredients on TP63 carcinoma cell.
In pancreatic cancer, the activities of inhibitory agents were investigated using docking, since the inhibition of TP63, which plays an important role in the spread of cancer with metastasis, in preventing the proliferation and proliferation of this type of cancer. It has been shown that the active ingredients in some plants used as traditional medicines have an inhibitory effect on this cancer type in preventing growth, reproduction and spread. These computational results guide experimental studies, preventing time and item loss; It is an important study in terms of choosing and using the right active substances.


Piriformospora indica colonization increases the growth, development, and herbivory resistance of sweet potato (Ipomoea batatas L.).
Piriformospora indica symbiosis promoted the growth and photosynthesis, and simultaneously enhanced the resistance against insect herbivory by regulating sporamin-dependent defense in sweet potato. Piriformospora indica (P. indica), a versatile endophytic fungus, promotes the growth and confers resistance against multiple stresses by root colonization in plant hosts. In this study, the effects of P. indica colonization on the growth, physiological change, and herbivore resistance of leaf-vegetable sweet potato cultivar were investigated. P. indica symbiosis significantly improved the biomass in both above- and under-ground parts of sweet potato plants. In comparison with the non-colonized plants, the content of photosynthetic pigments and the efficiency of photosynthesis were increased in P. indica-colonized sweet potato plants. Further investigation showed that the activity of catalase was enhanced in both leaves and roots of sweet potato plants after colonization, but ascorbate peroxidase, peroxidase, and superoxide dismutase were not enhanced. Furthermore, the interaction between P. indica and sweet potato plants also showed the biological function in jasmonic acid (JA)-mediated defense. The plants colonized by P. indica had greatly increased JA accumulation and defense gene expressions, including IbNAC1, IbbHLH3, IbpreproHypSys, and sporamin, leading to elevated trypsin inhibitory activity, which was consistent with a reduced Spodoptera litura performance when larvae fed on the leaves of P. indica-colonized sweet potato plants. The root symbiosis of P. indica is helpful for the plant promoting growth and development and has a strong function as resistance inducers against herbivore attack in sweet potato cultivation by regulating sporamin-dependent defense.


The British Pharmacological Society--winter meeting. Neuropharmacology. 6-8 January 1999, Brighton, UK.
The conference comprised a series of symposia on the general theme of aging, together with the usual disparate mixture of oral and poster sessions, covering the breadth of modern pharmacology. The major sponsor was Servier and the host institution was King's College, London. This report will encompass the major new neuropharmacological aspects of the meeting.


[Effect of immunostimulatory sequence on the expression of EOTAXIN of experimental allergic rhinitis].
To investigate the effect of CpG DNA applied intraperitoneally on the expression of EOTAXIN of allergic rhinitis model. CpG DNA was injected intraperitoneally to rat at the same time of the basic intraperitoneal immunized. The symptoms, expression of eosinophil (EOS) in nasal cavity and expression of EOTAXIN in the nasal mucosa were measured. After the CpG DNA was applied, the symptom scores, eosinophil (EOS) in nasal cavity and expression of EOTAXIN were weaken. There was great reverse effect of CpG DNA on the foundation of rat model of experimental allergic rhinitis, suppression of EOTAXIN may be one of the mechanisms.


Cyanide reduction by nitrogenase in intact cells of Rhodopseudomonas gelatinose Molisch.
Intact cells of Rhodopseudomonas gelatinosa, grown in N-free medium, reduce cyanide to methane and ammonia at a rate 5--8 times lower than acetylene. Cyanide reduction in whole cells is distinctly more sensitive to cyanide inhibition compared with enzyme preparations. These results are discussed in view of the exploitation of nitrogen-fixing photobacteria in the anaerobic detoxification of cyanide-containing wastewaters.


High level production of active streptokinase in Pichia pastoris fed-batch culture.
Streptokinase is a biological macromolecule involved in dissolution of fibrin blood clot and favourably used in various clinical applications. This protein is poorly expressed in soluble form due to its toxic effects on host physiology. The extracellular expression of recombinant streptokinase (SK) with and without 6xHis tag was obtained by cloning its gene under the α-mating factor signal sequence and alcohol inducible AOX1 promoter. Host-vector combinations were optimized to select a hyper producer. From shake flask optimization studies, a maximum expression of 582 mg/L of rSK (non-tagged) and 538 mg/L of rSK-His (His-tagged) protein was obtained when cells were induced at OD600 of 20. The high cell density fermentation increased the volumetric product concentration of rSK-His to a level of 4.25 g/L with a 7.9 folds increase from shake flask results. The specific product yield (YP/X) was 49.75 mg/g DCW along with a high volumetric productivity of 57.43 mg/L/h. The protein was predicted to have 15.43% α-helix and 26.43% β-sheet with tryptophan emission maxima of around 347 nm. The highest specific activity of rSK-His was 64,903 IU/mg with 1.48 folds purification whereas specific activity of rSK was 55,240 IU/mg with 1.22 folds purification.


Efficacy of diclazuril in the prevention and cure of intestinal and hepatic coccidiosis in rabbits.
The efficacy of diclazuril against intestinal and hepatic coccidiosis was studied in artificially infected rabbits. Prophylaxis against intestinal coccidiosis was evaluated using a mixed infection of Eimeria intestinalis, Eimeria magna and Eimeria perforans. Continuous medication in the feed at 1 p.p.m. was 100% effective in reducing oocyst output and faecal scores, and weight gain and feed efficiency were normal. Hepatic coccidiosis induced by Eimeria stiedai was prevented at 0.5 and 1 p.p.m. as shown by negative oocyst counts, normal liver weight, absence of liver lesions, and normal body-weight gain and feed efficiency. Medication at 1 p.p.m. for 7 consecutive days during the prepatent phase of hepatic coccidiosis resulted in large reductions in oocyst counts and lesion scores with a normal liver weight and growth performance. Diclazuril at 1 p.p.m. in the feed prevented both intestinal and hepatic coccidiosis in rabbits and can be advocated for safe mass medication.


Two new species of Leptus Latreille, 1796 (Trombidiformes: Erythraeidae) from the Canary Islands, parasitising Curculionidae (Insecta: Coleoptera), with new metrical data for some Leptus spp.
Leptus (Leptus) grancanaricus n. sp. and L. (L.) machadoi n. sp. are described from Gran Canaria and Fuerteventura (Canary Islands). They were collected from new hosts for the genus Leptus: Herpisticus guanarteme Machado and Laparocerus maxorata Machado (Coleoptera: Curculionidae). New metrical data for Leptus (Leptus) andae, L. (L.) akkus, L. (L.) hammameticus, L. (L.) horiacus and L. (L.) tammuzi based on examination on the type-material are provided.


The Musculoskeletal Physiotherapy of Australia position on pre-manipulative testing for the cervical spine.
The MPA undertook a survey of its members in 1997 to determine their compliance with and opinion of the APA Protocol for Pre-Manipulative Testing of the Cervical Spine (Magarey et al 2000a, Magarey et al submitted-a). As a result of that survey and a comprehensive literature review, the MPA developed a new set of guidelines for premanipulative procedures for the cervical spine (Magarey et al 2000b, Magarey et al submitted-b).


Malignant fibrous histiocytoma: outcome of tumours in the head and neck compared with those in the trunk and extremities.
Malignant fibrous histiocytoma is one of the commonest soft tissue sarcomas in adults, affecting, in order of frequency, the extremities, trunk and head and neck. We treated 131 patients with malignant fibrous histiocytoma by radical, wide, or marginal resection. Their mean age was 43 years, and there were 54 in the head and neck and 77 in the trunk and extremities. The extent of clearance of the tumour, local recurrence, and 5-year survival were studied in these two groups. In the head and neck group, local recurrences developed in 86% after marginal resection, 66% after wide resection and 27% after radical resection. The comparative figures in the trunk and extremities group were 75, 71 and 18%, respectively. The overall 5-year survival was 48% in the head and neck group and 77% in the trunk and extremities group (p=0.03). Repeat operations for recurrences of tumour offered a 'cure rate' of 23% in the head and neck group and 61% in the trunk and extremities group. Inadequate resection of the sarcoma in the head and neck was associated with a high incidence of local recurrence and a poor prognosis. Therefore, we suggest that the initial operation for sarcoma in the head and neck should be as radical as possible to reduce the chance of local recurrence and to improve the outcome.


Test-retest reproducibility of elbow goniometric measurements in a rigid double-blinded protocol: intervals for distinguishing between measurement error and clinical change.
Increasingly, goniometry of elbow motion is used for qualification of research results. Expression of reliability is in parameters not suitable for comparison of results. We modified Bland and Altman's method, resulting in the smallest detectable differences (SDDs). Two raters measured elbow excursions in 42 individuals (144 ratings per test person) with an electronic digital inclinometer in a classical test-retest crossover study design. The SDDs were 0 +/- 4.2 degrees for active extension; 0 +/- 8.2 degrees for active flexion, both without upper arm fixation; 0 +/- 6.3 degrees for active extension; 0 +/- 5.7 degrees for active flexion; 0 +/- 7.4 degrees for passive flexion with upper arm fixation; 0 +/- 10.1 degrees for active flexion with upper arm retroflexion; and 0 +/- 8.5 degrees and 0 +/- 10.8 degrees for active and passive range of motion. Differences smaller than these SDDs found in clinical or research settings are attributable to measurement error and do not indicate improvement.


Impact of a family-centred clinical care programme on short-term outcomes of very low-birth weight infants.
We evaluated the effects of a family-centred clinical care pathway and case management programme on short-term clinical outcome in a cohort of very low-birth weight (VLBW) infants. The programme, named NeoPAss, was developed at the Department of Neonatology Children's hospital Passau in 2013. Short-term outcomes of infants were compared to matched controls from the Bavarian neonatology surveillance database before (n = 111; 2008-2012) and after implementation (n = 170; 2014-2017). After implementation the rate of late-onset sepsis was significantly lower (2.5% vs. 10.7%, p = 0.005) and the length of stay was significantly shorter (VLBW 28 to 31 weeks' gestational age (GA) 47.5 vs. 53.1 days, p = 0.047; <28 weeks' GA 79.4 vs. 91.9 days, p = 0.007) in the intervention group compared to controls. Infants were discharged with significantly lower weight (mean 2351 vs. 2539 g, p = 0.013). There was no statistically significant difference in the rate of intraventricular haemorrhage (3.7% vs. 8.2%), necrotizing enterocolitis (0.6% vs. 1.9%) and bronchopulmonary dysplasia (0% vs. 6.9%). Our data confirm that of other studies demonstrating a beneficial effect of family-centred care programmes and provides evidence that structured parental involvement is not associated with increased risk of infection in a VLBW cohort.


Co-existence of two types of [Ca2+]i-inducing protease-activated receptors (PAR-1 and PAR-2) in rat astrocytes and C6 glioma cells.
In the nervous system serine proteases, like thrombin, are involved in developmental and repair processes, but serve also as extracellular signalling molecules, acting via protease-activated receptors. Cellular responses of glial cells to thrombin are transduced by proteolytic activation of the G protein-coupled thrombin receptor. A second member of the protease-activated receptor family, protease-activated receptor-2, is activated by trypsin. We assessed whether glial cells express protease-activated receptor-2 together with the thrombin receptor. By reverse transcriptase polymerase chain reaction and Ca2+ imaging studies we demonstrate that rat astrocytes and C6 glioma cells functionally express protease-activated receptor-2. Short-term stimulation of the glial cells with thrombin, thrombin receptor agonist peptide, trypsin and protease-activated receptor-2 activating peptide dose-dependently induced a transient rise of [Ca2+]i. In astrocytes omission of extracellular Ca2+ attenuated the amplitude of the [Ca2+]i transient induced by protease-activated receptor-stimulation. The decrease was strongest for the trypsin-evoked response and a reduction comparable in size (40%) was observed by pre-treatment with pertussis toxin. In astrocytes concentration-effect curves reveal that (i) the proteases had a higher potency than the respective receptor-activating peptides to induce a Ca2+ response, (ii) proteolytic activation of the receptors by thrombin or trypsin resulted in a double-sigmoidal concentration-effect curve, whereas non-proteolytic activation by receptor activating peptides resulted in a sigmoidal concentration dependence, and (iii) trypsin evoked a significantly greater Ca2+ response than thrombin. Preceding stimulation with trypsin nearly abolished the subsequent response to thrombin, whereas the trypsin-evoked Ca2+ transient was only slightly attenuated after a prior challenge with thrombin. This is the first study to show that neural cells (glial cells) functionally express both thrombin receptor and protease-activated receptor-2 coupled to the mobilization of intracellular calcium. Since calcium is the premier second messenger mediating adaptive changes within the CNS, these findings emphasize an important physiological function of serine proteases in mammalian brain.


Transcriptome analysis of Xanthomonas fragariae in strawberry leaves.
Xanthomonas fragariae is a quarantine bacterial pathogen that causes angular leaf spot on strawberry. The aim of our study was to analyse the mechanism of interaction of this bacterium with its host plant at the transcriptome level. For this purpose, mRNAs of X. fragariae growing in Wilbrink's medium and from infected strawberry cv. Elsanta plants were isolated and sequenced using the Illumina MiSeq platform. The expression profiles of the bacteria in Wilbrink's medium and in planta were very diverse. Of the 3939 CDSs recorded, 1995 had significantly different expression in planta (966 and 1029 genes were down- and upregulated, respectively). Among the genes showing increased expression in planta, those with eggNOG/COG (evolutionary genealogy of genes: Non-supervised Orthologous Groups/Cluster of Orthologous Groups) categories associated with bacterial cell motility, signal transduction, transport and metabolism of inorganic ions and carbohydrates and transcription were overrepresented. Among the genes with the most increased expression in planta, genes primarily associated with flagella synthesis and chemotaxis were found. It is also interesting to note that out of the 31 genes localized on a plasmid, 16 were expressed differently in planta, which may indicate their potential role in plant-pathogen interactions. Many genes with differentiated expression that were localized on chromosome and plasmid encode proteins of unknown function.


Genetic variability in white-tailed deer.
Genetic variability at 36 loci was studied in white-tailed deer (Odocoileus virginianus) populations, on the Savannah River Plant (SRP) in South Carolina. Mean multilocus heterozygosity (H), percentage of polymorphic loci (P) and average number of alleles per locus (A) for white-tailed deer were calculated and compared with values for white-tailed deer from several locations (SRP deer: H = 9.9 per cent, P0.05 = 30.6 per cent, A = 1.89; white-tailed deer overall: H = 10.4 per cent, P0.05 = 32.3 per cent, A = 1.94). Frequency distributions for single locus heterozygosity values (h) and the number of alleles per locus for white-tailed deer were found to be significantly different from those of mammals in general. Analysis of single-locus data based on quaternary structure and functional groups of proteins failed to demonstrate expected differences as predicted from the literature. White-tailed deer have a high level of heterozygosity, but they do not exhibit many of the life history and environmental characteristics associated with high heterozygosity in other animals.


Acute infantile gastroenteritis caused by rotavirus in Japan.
We had an outbreak of acute infantile gastroenteritis accompanied by milky-white stool (called Hakuri in Japanese) during the winters of 1976 and 1977. Stool specimens collected from 72 cases of Hakuri were studied by negative-staining electron microscopy. Rotavirus was detected with a very high frequency (89%). Rotavirus obtained from one of the patients was isolated and passaged in cultures of primary human embryonic kidney cells. Viral antigens could be detected in the cytoplasm of the cells by indirect immuno-fluorescence. The fluorescence-positive cells increased in number with repeated passage. Serum anti-viral activities in 11 patients were titrated by indirect immuno-fluorescence, using the cells infected with the passaged rotavirus. All 11 patients developed IgM responses in the convalescent phase. However, in 4 of the 11 patients, no IgG responses were detected even 2--3 weeks after the onset of illness. The reinfection which has occasionally be seen in our country may be related to these poor IgG responses.


Clinical aspects of urea cycle dysfunction and altered brain energy metabolism on modulation of glutamate receptors and transporters in acute and chronic hyperammonemia.
In living organisms, nitrogen arise primarily as ammonia (NH3) and ammonium (NH4(+)), which is a main component of the nucleic acid pool and proteins. Although nitrogen is essential for growth and maintenance in animals, but when the nitrogenous compounds exceeds the normal range which can quickly lead to toxicity and death. Urea cycle is the common pathway for the disposal of excess nitrogen through urea biosynthesis. Hyperammonemia is a consistent finding in many neurological disorders including congenital urea cycle disorders, reye's syndrome and acute liver failure leads to deleterious effects. Hyperammonemia and liver failure results in glutamatergic neurotransmission which contributes to the alteration in the function of the glutamate-nitric oxide-cGMP pathway, modulates the important cerebral process. Even though ammonia is essential for normal functioning of the central nervous system (CNS), in particular high concentrations of ammonia exposure to the brain leads to the alterations of glutamate transport by the transporters. Several glutamate transporters have been recognized in the central nervous system and each has a unique physiological property and distribution. The loss of glutamate transporter activity in brain during acute liver failure and hyperammonemia is allied with increased extracellular brain glutamate concentrations which may be conscientious for the cerebral edema and ultimately cell death.