A Generic Assay to Detect Aberrant ARSB Splicing and mRNA Degradation for the Molecular Diagnosis of MPS VI.
Identification and characterization of disease-associated variants in monogenic disorders is an important aspect of diagnosis, genetic counseling, prediction of disease severity, and development of therapy. However, the effects of disease-associated variants on pre-mRNA splicing and mRNA degradation are difficult to predict and often missed. Here we present a generic assay for unbiased identification and quantification of arylsulfatase B (ARSB) mRNA for molecular diagnosis of patients with mucopolysaccharidosis VI (MPS VI). We found that healthy control individuals have inefficient ARSB splicing because of natural skipping of exon 5 and inclusion of two pseudoexons in introns 5 and 6. Analyses of 12 MPS VI patients with 10 different genotypes resulted in identification of a 151-bp intron inclusion caused by the c.1142+2T>C variant and detection of low ARSB expression from alleles with the c.629A>G variant. A special case showed skipping of exon 4 and low ARSB expression. Although no disease-associated DNA variant could be identified in this patient, the molecular diagnosis could be made based on RNA. These results highlight the relevance of RNA-based analyses to establish a molecular diagnosis of MPS VI. We speculate that inefficient natural splicing of ARSB may be a target for therapy based on promotion of canonical splicing.


Double-balloon ERCP in patients who have undergone Roux-en-Y surgery: a case series.
ERCP is technically challenging in patients who have had a long-limb Roux-en-Y surgical procedure. The recent introduction of the double-balloon endoscope permits the examination of a much longer segment of the small bowel compared with a standard endoscope and may be used to perform ERCP in these patients. To report successful use of double-balloon ERCP in patients who have had a Roux-en-Y surgical procedure. Fourteen patients with a history of either Roux-en-Y gastric bypass weight-reduction surgery or Roux-en-Y pancreatobiliary surgery required diagnostic and/or therapeutic pancreatobiliary intervention. Case report. Double-balloon ERCP. Fourteen patients underwent a total of 20 ERCPs with the double-balloon endoscopy system. The ampulla was successfully reached in 85% of total cases (100% of patients who have had Roux-en-Y weight reduction surgery), with adequate cannulation of either the biliary or pancreatic duct in 80% (88% of patients for weight reduction). Therapeutic intervention, including stone removal, pancreatobiliary-duct dilation, sphincterotomy, stent placement, and removal of previously placed stents, was performed successfully in 6 cases. The mean age was 47 years old. The mean (+/- standard deviation) total duration of the procedure was 99 +/- 48 minutes. There were no immediate or short-term complications. The double-balloon endoscopy system permits diagnostic and therapeutic ERCP in patients who have had long-limb surgical procedures. Our experience demonstrated that this procedure is well tolerated, safe, and has a high success rate.


Fine structure of experimental canine gastric carcinoma, with special reference to signet ring cells.
Fine structure of poorly differentiated adenocarcinoma selectively induced in the canine stomach by a low concentration of N-ethyl-N'-nitro-N-nitrosoguanidine was studied with special reference to the so-called signet ring cells. These cells were electron microscopically classified into two groups: 1) mucin-containing cells and 2) intracellular microcyst cells. The cytoplasm of mucin-containing cells was packed with fused large and low electrondense secretory granules. In another type of signet ring cells, there were intracellular microcysts with microvilli on the internal surface. A comparison of canine and human signet ring cells revealed a close resemblance in the fine structure. Canine gastric carcinoma should be a good model for human gastric carcinoma therapeutics.


The role of TGF-β in bone metastasis: novel therapeutic perspectives.
The skeleton is a preferred site for cancer metastasis. These bone metastases cause dysregulated bone remodeling and the associated morbidity of fractures, pain, hypercalcemia and catastrophic nerve compression syndromes. Transforming growth factor-β (TGF-β) is stored in mineralized bone matrix, and released and activated by osteoclastic bone resorption. Once activated, TGF-β stimulates nearby metastatic tumor cells within the bone microenvironment to secrete factors that further drive osteolytic destruction of the bone. Therefore, TGF-β and its signaling constitute a critical component driving the feed-forward vicious cycle of cancer growth in bone. Moreover, additional pro-tumorigenic activities attributed to TGF-β include activation of epithelial-to-mesenchymal transition, increased tumor cell invasion, enhanced angiogenesis and various immunomodulatory properties. Blocking the TGF-β signaling pathway to interrupt this vicious cycle and manipulate the bone microenvironment offers a promising area for therapeutic intervention to decrease skeletal metastasis and normalize bone homeostatic mechanisms. In this review, preclinical and clinical data are evaluated for the potential use of TGF-β pathway inhibitors in clinical practice to treat bone metastases and its associated comorbidities.


Low levels of pancreatic elastase 1 in stools of preterm infants.
The amount of faecal pancreatic enzyme elastase 1 was significantly lower in 42 preterm newborns than in 12 full term babies at day 2 (89 (3-539) v 354 (52-600) microg/g, p<0.0007) and day 5 (164 (3-600) v 600 (158-600) microg/g, p<0.05) and correlated positively with total nutrient intake during the first week of life in preterm infants. This should probably be taken into account during early feeding.


Contrasting thermal sensitivity of spontaneously active A- and C-fibers in experimental nerve-end neuromas.
Injured afferent A- and C-fibers ending in experimental neuromas in the rat sciatic nerve generate a substantial spontaneous discharge. We show that for individual axons the rate and percent incidence of spontaneous discharge are sensitive to neuroma temperature. Within the range of 14-43 degrees C, firing rate of all of the myelinated fibers examined increased as temperature rose, and decreased as temperature fell. For fibers with a tonic rhythmic discharge pattern, Q10 averaged 1.64 at 34-42 degrees C. Some fibers that were initially silent began to fire as the neuroma was warmed, and some fibers active at baseline temperature fell silent when the neuroma was cooled. Unmyelinated fibers behaved quite differently, showing either no response to temperature changes (44% of fibers sampled), or an increase in discharge rate upon cooling (56%). These effects are probably not secondary to vascular changes, but rather reflect thermal sensitivity of the ectopic neuroma impulse generator sites. This thermal sensitivity may account for the aggravation of phantom limb pain and other neuralgias during cold weather (i.e., post-traumatic cold intolerance).


Trapidil-mediated inhibition of CNS remyelination results from reduced numbers and impaired differentiation of oligodendrocytes.
In a previous study, we described the inhibitory effects of the growth factor-antagonist, trapidil, on spontaneously occurring oligodendrocyte remyelination in the rat spinal cord following lysolecithin-induced demyelination [30]. The objective of the present study was to further investigate the mechanisms of trapidil-mediated impairment of remyelination and thus obtain greater insight into the steps at which growth factors may be involved in remyelination. To this end, an ultrastructural analysis of the cellular composition of lesions from control and trapidil-treated animals was undertaken. Demyelination was created in the dorsal funiculus of 6-week-old female rats by the injection of 1.0 microliter of 1% lysolecithin. The animals received daily intraperitoneal injections of trapidil (80 mg/kg) or saline for 21 days, beginning on the day of lesion induction. Quantitative electron microscopic examination of lesions from both groups of animals showed that trapidil-treated lesions had reduced numbers of oligodendrocytes (P = 0.02) with a higher relative proportion of immature phenotypes, but increased numbers of microglia (P = 0.0009) and dystrophic axons (P0.02). In addition, the numbers of myelin lamellae around remyelinated axons were fewer in trapidil-treated animals. These results suggest that trapidil-mediated impairment of CNS remyelination is due to a blockage of growth factor-mediated proliferation and/or recruitment of remyelinating cells. Furthermore, the presence of oligodendrocytes with a more immature phenotype and the decreased thickness of the myelin sheaths of remyelination in the trapidil-treated animals indicate an impairment of growth factor-mediated differentiation.


Asymmetry of congruency effects in spatial Stroop tasks can be eliminated.
Three experiments examined whether asymmetry in interference can be eliminated in spatial Stroop tasks. In Experiment 1, responding to arrows or location words written in Chinese and to their locations created spatial Stroop effects of similar sizes. In Experiment 2, responding to a location word embedded in an outline drawing of arrow did not yield a spatial Stroop effect, but responding to the arrow's direction did yield an effect. In Experiment 3, responding to a location word flanked by an arrow and to the arrow rather than the word produced similar sizes of spatial Stroop effects. These results show that asymmetry in spatial Stroop interference can be eliminated in some situations. Although aspects of the results are consistent with predictions of translation and dimensional overlap models, they are in closest agreement overall with an account in terms of the relative strengths of the relevant and irrelevant stimulus-response associations.


A Novel Nomogram for the Preoperative Prediction of Edmondson-Steiner Grade III-IV in Hepatocellular Carcinoma Patients.
Edmondson-Steiner (E-S) grade is a pathological indicator of the degree of hepatocellular carcinoma (HCC) differentiation, and E-S grade III-IV is a poor prognostic factor for HCC patients. Predicting poorly differentiated HCC has essential significance for clinical decision-making. Although some studies have developed predictive models based on magnetic resonance imaging (MRI) and radiomics, radiomic features that require specific software for analysis are impractical for clinical work. This study aims to develop a novel and user-friendly nomogram model to predict E-S grade III-IV. Medical data on patients meeting the inclusion criteria were obtained from the Nanjing Drum Tower Hospital HCC database (January 2020 to December 2022). Univariate analysis was used to screen for risk factors associated with E-S grade III-IV. A novel nomogram was established based on the subsequent multivariate logistic regression analysis. The performance of the established model was evaluated through diagnostic ability, calibration, and clinical benefits. Overall, 240 HCC patients were included in this study. Among them, 103 were highly differentiated (E-S grade I-II) HCC and 137 were poorly differentiated (E-S grade III-IV) HCC. A nomogram model that integrated alpha-fetoprotein (AFP), des-γ-carboxy prothrombin (DCP), hepatitis B virus surface antigen (HBsAg), hepatitis C virus antibodies (HCVAb), aspartate aminotransferase to lymphocyte ratio index (ALRI), and macrovascular invasion was established. The novel model had a good diagnostic performance with an area under the curve (AUC) value of 0.763. Meanwhile, the model had a diagnostic accuracy of 72.5%, a sensitivity of 78.1%, and a specificity of 65.1%. The calibration curve showed good calibration of the nomogram model (mean absolute error = 0.043), and the decision curve analysis (DCA) demonstrated that the clinical benefit was provided. Our developed nomogram model could successfully predict E-S grade III-IV in HCC patients, which may be helpful in clinical decision-making.


Age of onset of drug use and its association with DSM-IV drug abuse and dependence: results from the National Longitudinal Alcohol Epidemiologic Survey.
The purpose of this study was to examine the relationship between early onset drug use and the development of lifetime DSM-IV drug abuse and dependence using a representative sample of the U.S. population. Prevalences of lifetime drug abuse and dependence were estimated for each year of age of onset of drug use from ages 13 and younger to 21 and older for the overall sample of drug users by race and gender. Linear logistic analyses were conducted to assess the relationship between age of drug use onset and lifetime drug use disorders controlling for important covariates. The major finding of this study was that early onset drug use is a significant predictor of the subsequent development of drug abuse over the life course. Early onset drug use was also a significant predictor of the subsequent development of lifetime alcohol dependence among males, females, and nonblacks, but not among blacks. After adjusting for important model covariates, the likelihood of lifetime drug abuse and dependence among the total sample of lifetime drug users was reduced by 4% and 5% with each year drug use onset was delayed. Implications of these findings are discussed in terms of the importance of collecting national data on drug use, abuse and dependence and the need for further research and its integration with prevention efforts.


A novel regulatory pathway in granulosa cells, the LH/human chorionic gonadotropin-microRNA-125a-3p-Fyn pathway, is required for ovulation.
Granulosa cells support the developing oocytes and serve as transducers of the ovulatory stimulus induced by LH surge. Fyn kinase is expressed in granulosa cells, though its role in these cells has not been studied. In human embryonic kidney 293T cells, microRNA (miR)-125a-3p down-regulates Fyn expression, causing a decrease in cells' migratory ability. Our aim was to explore the role of miR-125a-3p and Fyn in granulosa cells toward ovulation, focusing on migration as a possible mechanism. We demonstrate expression of miR-125a-3p and Fyn in mouse mural granulosa cells of preovulatory follicles and miR-125a-3p-induced down-regulation of Fyn expression in a granulosa cell line (rat). Administration of human chorionic gonadotropin (hCG; LH analog) caused a 75% decrease in the in vivo miR-125a-3p:Fyn mRNA ratio, followed by a 2-fold increased migratory ability of mural granulosa cells. In the hCG-treated granulosa cell line, miR-125a-3p expression was decreased, followed by Fyn up-regulation and phosphorylation of focal adhesion kinase and paxillin, enabling cell migration. An in vivo interference with miR-125a-3p:Fyn mRNA ratio in granulosa cells by intrabursal injections of Fyn small interfering RNA or miR-125a-3p mimic caused a 33 or 55% decrease in the number of ovulated oocytes, respectively. These observations reveal a new regulatory pathway in mural granulosa cells under the regulation of LH/hCG. Modulation of cell migration may account for the significance of the LH/hCG-miR-125a-3p-Fyn pathway to ovulation.


Robotic Surgery of the Kidney, Bladder, and Prostate.
Robot-assisted surgery offers the advantages of a minimally invasive approach with greater technical ease and a shorter learning curve than pure laparoscopy. Fueled by the success of the robot-assisted laparoscopic prostatectomy, urologists are increasingly using the robotic platform for other advanced operations involving the kidney, ureters, bladder, and prostate. Robotic surgery has been shown to be safe and effective, with good perioperative, functional, and oncologic outcomes. Although cost continues to be a major concern regarding the use of robotic technology, improved efficiency and reduced hospital stays associated with the minimally invasive approach are allowing for better cost-effectiveness.


Simultaneous boron (B) removal and electricity generation from domestic wastewater using duckweed-based wastewater treatment reactors coupled with microbial fuel cell.
Boron removal from water environment is a critical issue for scientific spotlight because its removal from wastewater is difficult and costly with conventional treatment method. Herein, an innovative, cost effective and attractive method which depends on duckweed-based wastewater treatment systems coupled with microbial fuel cell reactor (DWWT-MFC) was investigated for B-polluted domestic wastewater treatment and simultaneous electricity generation for the first time in an eco-technological study. Lemna gibba L. was selected as a model duckweed species, and different reactors were also designed to identify which mechanisms are dominant for B removal in a DWWT-MFC reactor matrix. DWWT-MFC reactor achieved 71% B removal in experiment period, and the plant effect on B removal mechanisms in the reactor matrix was recorded as 37.7 ± 4.92% (F = 2.543, p < 0.05). However, supplementary aeration and microbial effects on B removal were determined as negligible. Average maximum voltage output was found as 1.47 V, and maximum power density was 34.8 mW/m2 at a current density of 43.9 mA/m2 with supplementary aeration. Moreover, DWWT-MFC reactor achieved 84%, 81% and 76% of COD, NH4+ and PO43- removal efficiencies, respectively. Moreover, L. gibba grew well in the anode chamber of DWWT-MFC with an average biomass yield of 218 ± 43 g/m2 and a total chlorophyll (a+b) concentration of 30.2 mg g-1, which indicates that anolyte environment was not toxic for L. gibba growth. Consequently, it can be suggested that environmental experts may use DWWT-MFC as an efficient removal method to treat B from domestic wastewater and to produce bioelectricity.


Effect of cephalosporins on fascial healing after celiotomy.
A 7 day course of either cefonicid or cefazolin significantly reduced mean wound breaking weight after midline celiotomy in Sprague-Dawley rats compared with control animals. This detrimental effect was not seen when each drug was administered as a single preoperative dose. Even a 3 day course of cefonicid was associated with a significant reduction in the weight required to disrupt a healing abdominal closure. An increased incidence of incisional hernias was also noted among animals treated for 7 days with cefonicid or cefazolin. Shorter antibiotic regimens were not associated with an increased frequency of incisional herniation.


Changes in the midgut diverticula epithelial cells of the European cave spider, Meta menardi, under controlled winter starvation.
The European cave spider, Meta menardi, is among the most common troglophile species inhabiting the cave entrance zone in Europe, where prey is scarce in winter. Spiders feed only if prey is available; otherwise, they are subjected to long-term winter starvation. We carried out a four-month winter starvation of M. menardi under controlled conditions to analyze ultrastructural changes in the midgut diverticula epithelial cells at the beginning, in the middle and at the end of the starvation period. We used light microscopy, TEM and quantified reserve lipids and glycogen. The midgut diverticula epithelium consisted of secretory cells, digestive cells and adipocytes. During starvation, gradual vacuolization of some digestive cells, and some necrotic digestive cells and adipocytes appeared. Autophagic structures, autophagosomes, autolysosomes and residual bodies were found in all three cell types. Spherites and the energy-reserve compounds were gradually exploited, until in some spherites only the membrane remained. Comparison between spring, autumn and winter starvation reveals that, during the growth period, M. menardi accumulate reserve compounds in spherites and protein granules, and energy-supplying lipids and glycogen, like many epigean, overwintering arthropods. In M. menardi, otherwise active all over the year, this is an adaptive response to the potential absence of prey in winter.


Substitutions at auxiliary operator O3 enhance repression by nitrate-responsive regulator NarL at synthetic lac control regions in Escherichia coli K-12.
We constructed monocopy lac operon control regions in which the operators O1-lac and O3-lac were replaced by NarL and NarP binding sites from the nirB or napF operon control regions. The results support the hypothesis that DNA-bound dimers of phospho-NarL can participate in higher-order cooperative interactions.


Post-mortem radiology in fetal and neonatal death: the diagnostic value of post-mortem MRI versus autopsy regarding non-cardiac thoracic and abdominal abnormalities.
To compare the diagnostic value and accuracy of post-mortem magnetic resonance imaging (PMMRI) and autopsy for non-cardiac thoracic and abdominal abnormalities in fetal death. This single-institution retrospective study included all consecutive cases of fetal and perinatal death between January 2015 and December 2021 for which PMMRI followed by autopsy was conducted. These cases comprised fetuses at >18 weeks of gestation and preterm and term neonates who lived for <24 h. All PMMRI and autopsy reports were re-assessed and scored for seven non-cardiac thoracic and 52 abdominal abnormalities, and concordance between autopsy and PMMRI findings was determined as the primary outcome. Eighty cases were included in this study. Fetal loss was caused by termination of pregnancy in 80% of cases. Further, the mean gestational age was 166 days (23 weeks and 5 days, range 126-283 days). The concordance between PMMRI and autopsy for non-cardiac thoracic and abdominal abnormalities was 83.1% (95% confidence interval [CI] 71.3-83.3) and 76.3% (95% CI 65.8-84.2%), respectively, with a substantial and moderate strength of agreement (Cohen's kappa = 0.63 and 0.51 respectively). PMMRI exhibited good overall diagnostic value for non-cardiac thoracic and abdominal abnormalities, specifically large structural abnormalities. PMMRI may offer parents and physicians a valuable addition to autopsy for the detection of non-cardiac thoracic and abdominal abnormalities, or even an alternative option when parents do not consent to autopsy.


Opioid receptor stimulation does not affect cellular hypoxia-induced gene responses mediated by hypoxia-inducible factor 1 in cultured cell lines.
Hypoxia induces a series of adaptive physiological responses including gene inductions. Hypoxia-inducible factor 1 (HIF-1) is a master transcription factor that regulates hypoxia-induced gene expression to maintain homeostasis in the living body. Opioids are potent analgesic agents that are widely used in clinical practice. Therefore, we investigated the effect of opioids on HIF-1 activity. SH-SY5Y human neuronal cells, which express opioid receptors intrinsically, were cultured under 1% or 20% O2 conditions with or without treatment by DAGO, DPDPE, or U-50488, which are the selective agonists of mu-, delta-, and kappa-opioid receptors, respectively. Expression of subunits of HIF-1, HIF-1alpha, and HIF-1beta were examined by Western blot using specific antibodies. Expression of the HIF-1-dependent gene were investigated by reporter assay. None of the selective agonists of opioid receptors tested affected HIF-1 activation by hypoxia. Therefore, it is suggested that opioid receptor-mediated signals do not affect HIF-1-dependent cellular hypoxia-induced gene responses.


Epithelial-to-mesenchymal transition in FHC-silenced cells: the role of CXCR4/CXCL12 axis.
Ferritin plays a central role in the intracellular iron metabolism; the molecule is a nanocage of 24 subunits of the heavy and light types. The heavy subunit (FHC) is provided of a ferroxidase activity and thus performs the key transformation of iron in a non-toxic form. Recently, it has been shown that FHC is also involved in additional not iron-related critical pathways including, among the others, p53 regulation, modulation of oncomiRNAs expression and chemokine signalling. Epithelial to mesenchymal transition (EMT) is a cellular mechanism by which the cell acquires a fibroblast-like phenotype along with a decreased adhesion and augmented motility. In this work we have focused our attention on the role of the FHC on EMT induction in the human cell lines MCF-7 and H460 to elucidate the underlying molecular mechanisms. Targeted silencing of the FHC was performed by lentiviral-driven shRNA strategy. Reconstitution of the FHC gene product was obtained by full length FHC cDNA transfection with Lipofectamine 2000. MTT and cell count assays were used to evaluate cell viability and proliferation; cell migration capability was assayed by the wound-healing assay and transwell strategy. Quantification of the CXCR4 surface expression was performed by flow cytometry. Experimental data indicated that FHC-silenced MCF-7 and H460 cells (MCF-7shFHC, H460shFHC) acquire a mesenchymal phenotype, accompanied by a significant enhancement of their migratory and proliferative capacity. This shift is coupled to an increase in ROS production and by an activation of the CXCR4/CXCL12 signalling pathway. We present experimental data indicating that the cytosolic increase in ROS levels is responsible for the enhanced proliferation of FHC-silenced cells, while the higher migration rate is attributable to a dysregulation of the CXCR4/CXCL12 axis. Our findings indicate that induction of EMT, increased migration and survival depend, in MCF-7 and H460 cells, on the release of FHC control on two pathways, namely the iron/ROS metabolism and CXCR4/CXCL12 axis. Besides constituting a further confirmation of the multifunctional nature of FHC, this data also suggest that the analysis of FHC amount/function might be an important additional tool to predict tumor aggressiveness.


Neuromuscular complications of severe COVID-19 in paediatric patients: Medium-term follow-up.
Neuromuscular complications in paediatric patients with severe coronavirus disease 2019 (COVID-19) are poorly characterised. However, adult patients with severe COVID-19 reportedly present with frequent neuromuscular complications that mainly include critical illness polyneuropathy (CIP), critical illness myopathy (CIM), and focal neuropathies. We examined the records of all paediatric patients with severe COVID-19 who were mechanically ventilated and experienced neuromuscular complications from our single tertiary centre between March 2020 and August 2021. During this period, 4/36 (11%) patients admitted to the paediatric ICU who were mechanically ventilated experienced neuromuscular complications (one CIM, two focal neuropathies, and one CIP associated with plexopathy). In three of them, the gamma genetic variant of SARS-CoV-2 was identified. At the 4-5 month follow-up, three of our patients exhibited slight clinical improvement. We conclude that paediatric patients with severe COVID-19 may present neuromuscular complications similar to adults (11%), and their medium-term prognosis seems unfavourable.


Redefining and capturing pre-analytic deficiencies in an anatomical pathology laboratory: a quality improvement initiative.
Pre-analytical deficiencies (PADs) are a major source of errors in anatomical pathology, accounting for about 70% of laboratory deficiencies. These can lead to incorrect diagnoses, delayed treatments, and increased healthcare costs. As part of a quality improvement initiative, we retrospectively identified and characterized 237 PADs documented over a 1-year period in a tertiary care academic center. The most common PADs were errors in specimen procurement (56%), handling of samples within the lab (16%), accessioning (10%), incomplete requisitions (9%), and transportation-related issues (7%). Strategies were then devised to mitigate these errors. Categorization of pre- and intra-laboratory PADs was refined into eight categories (collection, requisition, specimen container, transportation, receiving, accessioning, preparation, and communications) in the laboratory information system. Mandatory PAD documentation was implemented for accessioning staff. Post-implementation, prospective analysis identified that the most common PADs were related to surgical requisitions (75%). Among these, missing ordering physician's signature was the most common, accounting for 67.7% of requisition-related PADs and 50.8% of all PADs. Other common PADs included incomplete information of specimens, clinical information, patient information, physician information, source location, collection time, incorrect requisition forms, and illegible handwritten information. This study highlights the importance of identifying and addressing PADs in the anatomical pathology laboratory setting as well as the potential benefits of implementing standardized documentation and quality improvement processes to address these deficiencies.


In vitro gene expression for the localization of antigenic determinants: application to the E. coli tryptophan synthase beta 2 subunit.
Gene expression of the beta subunit of E. coli tryptophan synthase in an E. coli cell-free transcription-translation system proceeds by pauses and produces a discrete but quite continuous pattern of nascent chains starting from the N terminus and ranging in size up to the 44 kDa end product corresponding to the completed beta chains. Using specific immunoadsorption of [35S]Met radiolabelled nascent chains by different monoclonal antibodies directed against the beta 2 subunit of E. coli tryptophan synthase, the size of the smallest N-terminal fragment reacting with each antibody has been determined by SDS electrophoretic analysis of the immunoadsorbed polypeptides. The immunoadsorption assay is performed in solution under conditions avoiding the usual drawbacks of solid phase immunoassay. This approach, in combination with the results obtained with a DNA fragment library permitted us to localize the antigenic determinants recognized by the monoclonal antibodies. The proposed method could help to localize rapidly the C-terminal boundary of an epitope, before starting systematic and precise mapping by other approaches. Moreover, the method described may be of general interest for the rapid production of a large set of C-terminal truncated polypeptides for studies of antigen-antibody recognition.


Assessment of extent of breast cancer: comparison between digital breast tomosynthesis and full-field digital mammography.
To compare the accuracy of digital breast tomosynthesis (DBT) and full-field digital mammography (FFDM) in preoperative assessment of local extent of breast cancer. Lesion sizes of breast cancers on DBT and FFDM images were independently evaluated by breast radiologists. Each lesion was flagged as either mis-sized or not depending on whether the assessment of size at imaging was within 1 cm of the lesion size at surgery. Additional analyses were made by mammographic parenchymal density and by lesion size, using 2 cm as the boundary to separate the two subgroups. Statistical comparisons were performed using a repeated measures linear model on the percent mis-sized. P-values < 0.05 were considered statistically significant. The dataset included 173 malignant breast lesions (mean size 23.8 mm, 43% of lesions were ≤2 cm in size) in 169 patients, two-thirds of which had heterogeneously or extremely dense breasts. Overall, the percentage of lesions mis-sized at DBT was significantly lower than at FFDM (19% versus 29%, p = 0.003). There was significantly less mis-sizing at DBT in both heterogeneously dense breasts (11.1% difference between DBT and FFDM, p = 0.016) and extremely dense breasts (15.8% difference, p = 0.024). DBT also had significantly less mis-sizing than FFDM in the subgroup of lesions that were ≤2 cm in size (14.7% difference, p = 0.005). DBT was significantly superior to FFDM for the evaluation of lesion size overall, and specifically for small lesions and for lesions in dense breasts. The superiority of DBT versus FFDM increased with parenchymal density.


Expression of GDF-9, BMP-15 and their receptors in mammalian ovary follicles.
The synergetic process of folliculogenesis is mainly regulated by GDF-9 and BMP-15 as well as their receptors, such as BMPR2, TβR1 and BMPR1B. Expressions of these factors and the receptors are significant different among species. This study was designed to detect expression of GDF-9, BMP-15 and their receptors in mouse, porcine and human healthy follicles by immunohistochemistry. Three ages of human ovary were studied according to ovarian developmental schedule, i.e. gestational week (GW) 16, puberty (14 year-old) and adult (40 year-old). The results showed that both GDF-9 and BMP-15 were detectable in oocytes from primary follicles onward, besides, BMP-15 also presented in granulosa cells (GCs) and follicular follicle of mature follicles in mouse. However, they were maintained in oocytes and GCs from primordial to mature follicles in porcine except that GDF-9 was undetectable in GCs of mature follicles. For human ovary, GDF-9 presented in oocytes of primordial follicles in all samples, whereas BMP-15 was only observed in primordial follicle of adult ovary. Receptors, BMPR2, TβR1 and BMPR1B were found in oocytes and GCs of all follicles in mouse and porcine. In human, they were stained in oocytes from primordial follices but BMPR1B was not expressed in pubertal primordial follicles. Furthermore, we found that GDF-9, BMP-15 and three receptors distributed in adult corpus lutea. Collectively, our studies suggested that GDF-9, BMP-15 and their receptors might correlate with primordial follicular recruitment in pig and human. Positive expression of the receptors (BMPR2, TβR1 and BMPR1B)in primordial follicles of mouse ovaries indicated that these receptors might interact with others ligands besides GDF-9 and BMP-15 to regulate primordial follicular activity in mouse. Moreover, presence of GDF-9 in oocytes and BMP-15 in oocytes and GCs of mature follicles from mice and porcine elucidated coordinated roles of GDF-9 and BMP-15 in cumulus oophorus expansion. Additionally, expression of these factors in adult human corpus lutea suggested they play roles in corpus luteum activity.


IL-25 exacerbates autoimmune aortitis in IL-1 receptor antagonist-deficient mice.
IL-25, a member of the IL-17 family of cytokines, is known to enhance type 2 immune responses, but suppress type 3 (IL-17A)-mediated immune responses. Mice deficient in IL-1 receptor antagonist (Il1rn-/- mice) have excessive IL-1 signaling, resulting in spontaneous development of IL-1-, TNF- and IL-17A-dependent aortitis. We found that expression of II25 mRNA was increased in the aortae of Il1rn-/- mice, suggesting that IL-25 may suppress development of IL-1-, TNF- and IL-17A-dependent aortitis in Il1rn-/- mice by inhibiting type 3-mediated immune responses. However, we unexpectedly found that Il25-/-Il1rn-/- mice showed attenuated development of aortitis, accompanied by reduced accumulation of inflammatory cells such as dendritic cells, macrophages and neutrophils and reduced mRNA expression of Il17a and Tnfa-but not Il4 or Il13-in local lesions compared with Il1rn-/- mice. Tissue-, but not immune cell-, derived IL-25 was crucial for development of aortitis. IL-25 enhanced IL-1β and TNF production by IL-25 receptor-expressing dendritic cells and macrophages, respectively, at inflammatory sites of aortae of Il1rn-/- mice, contributing to exacerbation of development of IL-1-, TNF- and IL-17A-dependent aortitis in those mice. Our findings suggest that neutralization of IL-25 may be a potential therapeutic target for aortitis.


Critical analysis of urethroplasty for male anterior urethral stricture: a single-center experience.
Urethroplasty is the most effective treatment method for anterior urethral strictures, which constitute an important health concern for males. This study aims to investigate factors that may predict treatment failure following urethroplasty for anterior urethral strictures and evaluate outcomes of different urethroplasty techniques. This retrospective study utilized univariate and multivariate analyses to identify factors affecting treatment success following different urethroplasty techniques performed by a single surgeon on male patients with anterior urethral strictures. Surgical outcomes of different urethroplasty techniques were evaluated individually. Urethroplasty procedures performed on 244 patients with a mean age of 54 years and a mean stricture length of 4.7 cm produced a success rate of 79.1%. Multivariate analyses identified stricture length and number of previous internal urethrotomy procedures as factors predicting urethroplasty success (HR 1.265, 95% CI 1.129-1.416, p < 0.001 and HR 1.188, 95% CI 1.038-1.361, p = 0.013, respectively). The eight surgical techniques used by the surgeon produced success rates that varied between 50% and 86.2%. Urethroplasty can offer satisfactory outcomes for anterior urethral strictures. Accordingly, a longer stricture length and greater number of previous internal urethrotomy procedures were associated with greater risk for urethroplasty failure. Moreover, urethroplasty techniques must be selected based on multifactorial evaluation and performed at experienced centers.


A chemically inert drug can stimulate T cells in vitro by their T cell receptor in non-sensitised individuals.
Drugs can interact with T cell receptors (TCR) after binding to peptide-MHC structures. This binding may involve the formation of a stable, covalent bond between a chemically reactive drug and MHC or the peptide embedded within. Alternatively, if the drug is chemically inert, the binding may be non-covalent and readily reversible. Both types of drug presentation account for a substantial number of adverse side effects to drugs. Presently no tests are available to predict the ability of chemically inert drugs to stimulate an immune response. Here we present data on the successful induction of a primary T cell immune response in vitro against a chemically inert drug using blood from healthy individuals, previously not exposed to the drug. Blood lymphocytes were stimulated by the chemically inert drug sulfamethoxazole and the protein-reactive drug-metabolite sulfamethoxazole-nitroso in the presence of IL-2. 9/10 individuals reacted in response to sulfamethoxazole-nitroso, but only three reacted to the chemically inert compound sulfamethoxazole. Drug reactive T cells could be detected after 14-35 days of cell culture by drug-specific proliferation or cytotoxicity, which was MHC-restricted. These cells were CD4, CD8 positive or CD4/CD8 double positive and T cell clones generated secreted Th0 type cytokines. Drug interaction lead to down-regulation of specific TCR. These data confirm the ability of chemically inert drugs to stimulate certain T cells by their TCR and may provide the opportunity to screen new drugs for their ability to interact with TCRs.


Age-dependent effects on sensory axonal excitability in normal mice.
Serial recordings were performed to measure sensory excitability in peripheral nerves and elucidate age-dependent changes in neuronal ion currents in the peripheral sensory nervous system. The threshold tracking technique was used to measure multiple excitability indices in the tail sensory nerves of five normal male mice at four time points (6, 10, 14, and 19 weeks of age). A separate group of four mice was also measured at 43 weeks and at 60 weeks of age. Maturation was accompanied by an increase in early hyperpolarization and superexcitability at 10 weeks. At 60 weeks, the hyperpolarizing electrotonus shifted downward, while superexcitability became greater and subexcitability (double stimuli) decreased. Computer modeling showed that the most notable age-related interval changes in excitability parameters were Barrett-Barrett, H, and slow K(+) conductances. Understanding age-related changes in the excitability of sensory axons may provide a platform for understanding age-dependent sensory symptoms and developing age-specific channel-targeting therapies.


The building blocks of a 'Liveable Neighbourhood': Identifying the key performance indicators for walking of an operational planning policy in Perth, Western Australia.
Planning policy makers are requesting clearer guidance on the key design features required to build neighbourhoods that promote active living. Using a backwards stepwise elimination procedure (logistic regression with generalised estimating equations adjusting for demographic characteristics, self-selection factors, stage of construction and scale of development) this study identified specific design features (n=16) from an operational planning policy ("Liveable Neighbourhoods") that showed the strongest associations with walking behaviours (measured using the Neighbourhood Physical Activity Questionnaire). The interacting effects of design features on walking behaviours were also investigated. The urban design features identified were grouped into the "building blocks of a Liveable Neighbourhood", reflecting the scale, importance and sequencing of the design and implementation phases required to create walkable, pedestrian friendly developments.


Effects of oligomeric proanthocyanidins on quality of boar semen during liquid preservation at 17 °C.
The use of antioxidants is an important aspect in the preservation of boar semen. Oligomeric proanthocyanidins (OPC) are an effective natural antioxidants that scavenges free radicals and reactive oxygen species (ROS). This study was designed to investigate the antioxidative effect of OPC on boar semen quality during liquid preservation at 17 °C. The effects of different concentrations of OPC on sperm quality variables, antioxidant effects and fertility were analyzed in this experiment. Semen collected from six Guanzhong-Black boars and was diluted with Beltsville thawing solution (BTS). During the process of liquid preservation at 17 °C, the variables assessed were measured and analyzed every 24 h. The addition of OPC improved boar sperm motility, acrosome integrity, membrane integrity, and mitochondrial membrane potential as compared with that of the control group (P < 0.05). Meanwhile, malondialdehyde content (MDA) and ROS content was less after adding OPC, thereby improving the total antioxidant capacity (T-AOC) (P < 0.05). Different concentrations of OPC have different degrees of protective effects on boar semen quality. The results indicate that 50 μg/mL of OPC was the optimum concentration, and that the conception rate, litter size, and survival rate increased at this concentration as compared with that of the control group (P < 0.05). In summary, the addition of OPC to BTS diluents can improve the quality of boar semen at 17 °C during liquid preservation. Further research is needed to understand the mechanism by which OPC provides protection to boar semen during preservation in vitro.


The 26S proteasome system degrades the ERM transcription factor and regulates its transcription-enhancing activity.
ERM is a member of the ETS transcription factor family. High levels of the corresponding mRNA are detected in a variety of human breast cancer cell lines, as well as in aggressive human breast tumors. As ERM protein is almost undetectable in these cells, high degradation of this transcription factor has been postulated. Here we have investigated whether ERM degradation might depend on the proteasome pathway. We show that endogenous and ectopically expressed ERM protein is short-lived protein and undergoes proteasome-dependent degradation. Deletion mutagenesis studies indicate that the 61 C-terminal amino acids of ERM are critical for its proteolysis and serve as a degradation signal. Although ERM conjugates with ubiquitin, this post-translational modification does not depend on the C-terminal domain. We have used an Ets-responsive ICAM-1 reporter plasmid to show that the ubiquitin-proteasome pathway can affect transcriptional function of ERM. Thus, ERM is subject to degradation via the 26S proteasome pathway, and this pathway probably plays an important role in regulating ERM transcriptional activity.


Genomic structure of the proopiomelanocortin gene and expression during acute low-water stress in channel catfish.
Proopiomelanocortin (POMC) is an important gene involved in the stress response of the hypothalamic-pituitary-adrenal axis. It is a precursor of several peptide hormones including adrenocorticotropic hormone, melanocyte stimulating hormones, and beta-endorphin. Our study aims to determine genomic structure and expression of POMC gene during temporal stress in channel catfish (Ictalurus punctatus). The catfish POMC gene consisting of three exons and two introns has a similar structural organization to that of other species. The catfish and mammalian POMC promoters do not exhibit regions of conservation except that of one TATA box. Genomic Southern blot analysis indicated POMC is present as a single copy gene in the catfish genome. Real-time PCR allowed us to monitor temporal expression of the POMC mRNA in catfish pituitary during low-water stress. Plasma cortisol concentrations were also measured as an indicator of stress. Within 15 min after the onset of low-water stress, POMC mRNA expression was elevated 1.87-fold above the control value. The POMC mRNA level had declined after 30 min (1.29-fold) and 1h (1.1-fold) at which time stress was removed. After 1h recovery, a significant increase in the POMC mRNA expression was detected (2.44-fold, P<0.05) followed by a decline 2h later (1.52-fold) when the experiment was terminated. Plasma cortisol levels in stressed fish were significantly above the cortisol levels in control fish during stress application (t=15 min, t=30 min, and t=1h, P<0.05), which then returned to normal during recovery. We conclude that POMC and cortisol are both involved in the low-water stress response during which cortisol may serve as a negative regulator of POMC expression in catfish.


Structural and mechanistic basis of the interaction between a pharmacological chaperone and human phenylalanine hydroxylase.
Not without a chaperone: Pharmacological chaperones are designed to bind and ideally stabilise their target protein. Here, we elucidate the molecular mechanism of a potential pharmacological chaperone to treat phenylketonuria. The crystal structure of human phenylalanine hydroxylase with compound IV may help in the rational design of more efficient compounds to treat this disease.


Studying the human immunome: the complexity of comprehensive leukocyte immunophenotyping.
A comprehensive study of the cellular components of the immune system requires both deep and broad immunophenotyping of numerous cell populations in an efficient and practical manner. In this chapter, we describe the technical aspects of studying the human immunome using high-dimensional (15 color) fluorescence-based immunophenotyping. We focus on the technical aspects of polychromatic flow cytometry and the initial stages in developing a panel for comprehensive leukocyte immunophenotyping (CLIP). We also briefly discuss how this panel is being used and the challenges of encyclopedic analysis of these rich data sets.


Bumblebee visits to Impatiens spp.: pattern and efficiency.
Three Japanese species of Impatiens, which secrete nectar continuously in long spurs, were visited by Bombus diversus workers consecutively throughout the day. B. diversus workers showed characteristic patterns of behavior in flower use, flower choice, and patch departure. (1) Bumblebees stayed longer on a flower which had been unvisited for a while than on a flower which had been visited recently. (2) Bumblebees preferred visiting flowers which had been unvisited for a while to visiting those which had been visited recently, and to visiting those which had been unvisited for a long period. (3) Bumblebees had a higher probability of leaving a patch after they had stayed on a flower for a short period than after they had stayed for a longer period. The bumblebees appeared to perceive both remotely and proximately chemical cues deposited by other foraging individuals, which indicated nectar rewards in a flower, and thus obtained a higher nectar intake than the mean amount of nectar left in a flower.


Effects of epidermal growth factor and insulin-like growth factor I on the levels of mRNA encoding aromatase cytochrome P-450 of human ovarian granulosa cells.
The effects of growth factors to regulate the activity of aromatase, as well as the synthesis of aromatase cytochrome P-450 (P-450AROM) have been studied in human ovarian granulosa cells obtained from women undergoing oocyte retrieval. Insulin-like growth factor I (IGF-I) increased aromatase activity as well as the synthesis of P-450AROM, in a concentration-dependent fashion. The levels of hybridizable mRNA species encoding cytochrome P-450AROM were also increased with IGF-I treatment. By contrast, epidermal growth factor (EGF) had no effect on these parameters when added alone, but markedly inhibited the action of follicle-stimulating hormone (FSH) to stimulate aromatase activity, and the synthesis of cytochrome P-450AROM, as well as its ability to increase the levels of mRNA encoding the enzyme. It is concluded that these growth factors have opposite effects on aromatase activity, and that these actions reflect, in part, changes in the synthesis of cytochrome P-450AROM, which in turn are the consequence of changes in the levels of mRNA encoding this enzyme.


Validation of the Spanish Version of the Quality of Dying and Death Questionnaire (QODD-ESP) in a Home-Based Cancer Palliative Care Program and Development of the QODD-ESP-12.
Improving quality of death (QOD) is a key goal in palliative care (PC). To our knowledge, no instruments to measure QOD have been validated in Spanish. The goals of this study were to validate the Spanish version of the quality of dying and death (QODD) questionnaire and to develop and validate a shortened version of this instrument by phone interview. We enrolled caregivers (CGs) of consecutive deceased cancer patients who participated in a single PC clinic. CGs were contacted by phone between 4 and 12 weeks after patients' death and completed the Spanish QODD (QODD-ESP). A question assessing quality of life during last week of life was included. A 12-item QODD (QODD-ESP-12) was developed. Reliability, convergent validity, and construct validity were estimated for both versions. About 150 (50%) of 302 CGs completed the QODD-ESP. Patient's mean age (SD) was 67 (14); 71 (47%) were females, and 131 (87%) died at home. CGs' mean age (SD) was 51 (13); 128 (85%) were females. Mean QODD-ESP score was 69 (range 35-96). Kaiser-Meyer-Olkin measure of sampling adequacy was 0.322, not supporting the use of factorial analysis to assess the existence of an underlying construct. Mean QODD-ESP-12 score was 69 (range 31-97). Correlation with last week quality of life was 0.306 (P < 0.01). Confirmatory factorial analysis of QODD-ESP-12 showed that data fitted well Downey's four factors; Chi-square test = 6.32 (degrees of freedom = 60), P = 0.394 comparative fit index = 0.988; Tucker-Lewis Index = 0.987, and root mean square error of approximation = 0.016 (95% CI 0-0.052). QODD-ESP-12 is a reliable and valid instrument with good psychometric properties and can be used to assess QOD in a Spanish-speaking cancer PC population by phone interview.


Effects of epidural anesthesia on catecholamines, renin activity, and vasopressin changes induced by tilt in elderly men.
Mean arterial pressure, heart rate, plasma catecholamines, renin activity, and vasopressin changes induced by a 30-degree head-up tilt were studied before and during epidural anesthesia with bupivacaine in eight elderly patients (ages 58-82 yr). The tilt performed before epidural anesthesia did not modify mean arterial pressure, heart rate, plasma catecholamines, renin activity, and vasopressin at 5 and 15 min. During epidural anesthesia, the superior level of analgesia ranged from T4 to T10. Epidural anesthesia induced significant (P less than 0.05) decreases from control values in mean arterial pressure and plasma norepinephrine (from 85 +/- 6 to 67 +/- 8 mmHg and from 600 +/- 108 to 307 +/- 77 pg/ml, respectively, mean +/- SEM) without significant changes in heart rate, plasma epinephrine, renin activity, and vasopressin. However 5 and 15 min after tilt, significant decreases from pretilt values were measured in mean arterial pressure (from 67 +/- 8 to 57 +/- 6 and 55 +/- 6 mmHg, respectively) and in heart rate (from 70 +/- 8 to 63 +/- 7 and 62 +/- 7 beats/min). Simultaneously, an increase in plasma vasopressin (from 14.8 +/- 5.5 to 36.2 +/- 10.3 and 40.0 +/- 10.5 pg/ml) was recorded, whereas plasma norepinephrine and epinephrine remained unchanged. Posttilt plasma renin activity values at 5 and 15 min were increased significantly when compared with the preepidural values (2,752 +/- 1,168, 2,410 +/- 1,214 and 713 +/- 190 pg X ml-1 X h-1, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)


Schistosoma mansoni: praziquantel-induced changes to the female reproductive system.
The long-term in vivo effects of a single subcurative dose (200 mg/kg body wt of mouse) of praziquantel on the ultrastructure of the female reproductive system of Schistosoma mansoni were investigated. Morphological changes in the structure of both the vitelline gland and the ovary were apparent within 24 hr post-treatment, and lead to a partial or complete regression of both organ systems. Associated with this regression was a cessation of egg production. In surviving, paired females, irrespective of the initial severity of the drug-induced damage, both the vitelline gland and the ovary completely redeveloped and lead eventually to a resumption of egg production. In contrast, in unpaired, previously mature females the reproductive system also regressed but did not redevelop. In these cases, although the initial changes in the reproductive system were the result of drug action, the long-term regressive changes were due to discontinued male stimulation.


The use of the Scar Cosmesis Assessment and rating scale to evaluate the cosmetic outcomes of totally thoracoscopic cardiac surgery.
Conventional median sternotomy is widely used in cardiac surgery, while thoracoscopic cardiac surgery, which is considered to have aesthetic advantages, is being performed increasingly more often in China because patients' requests for minimally invasive procedures yielding aesthetically pleasing results have significantly increased. Few studies have been conducted to assess surgical scars after cardiac surgery. Compared to the median sternotomy approach, multiple-incision totally thoracoscopic cardiac surgery requires smaller but numerous and scattered incisions. In addition to two working ports on the upper and lower margins of the right breast, an inguinal incision and an axillary incision are made. Therefore, does totally thoracoscopic cardiac surgery truly have aesthetic advantages? This study has the following objectives: (a) to compare median sternotomy cardiac surgery and total thoracoscopic cardiac surgery in terms of the long-term cosmetic outcomes of post-operative scars and (b) to evaluate the effectiveness of the Scar Cosmesis Assessment and Rating scale in combination with the numeric rating scale in the assessment of surgical scars after cardiac surgery. Consecutive patients who visited our institution from January 2019 to May 2019 for cardiac surgery via median sternotomy or the totally thoracoscopic approach and followed up for at least one year were included. Inter-rater reliability, internal consistency and convergent validity were evaluated for the Scar Cosmesis Assessment and Rating scale and the numeric rating scale. Clinical characteristics and the scores of the two scales were compared between the two groups using Student's t test or the Mann-Whitney U test. Thirty-one patients underwent cardiac surgery via the totally thoracoscopic approach, and 42 patients underwent cardiac surgery via the median sternotomy approach. No significant differences were found in the demographic or clinical data between the two groups. The validity and reliability of the two scales were satisfactory. For the Scar Cosmesis Assessment and Rating scale, the median sternotomy group scored statistically significantly higher than did the totally thoracoscopic group on the "overall impression" and "patient question" subscales (P < 0.05). The overall scores of the Scar Cosmesis Assessment and Rating scale and numeric rating scale were statistically significantly different (P < 0.05). The Scar Cosmesis Assessment and Rating scale in combination with the numeric rating scale is an effective tool for the assessment of scar aesthetics after cardiac surgery. Surgical scars of totally thoracoscopic cardiac surgery can yield desirable cosmetic outcomes in Chinese individuals, especially in susceptible individuals with a high risk of keloid and hypertrophic scars. Patients with appropriate indications can undergo cardiac surgery with the totally thoracoscopic approach and exhibit a satisfactory scar appearance.


Efficacy of transcutaneous posterior tibial nerve stimulation in children with functional voiding disorders.
To assess the efficacy of transcutaneous posterior tibial nerve stimulation (TPTNS) on functional voiding disorder (FVD) and investigate the utility of urine biomarkers (UBs: nerve growth factor, transforming growth factor-beta 1, and tissue inhibitor of metalloproteinases 2) in diagnosis and follow-up. A total of 44 children were included to this randomized controlled trial prospectively. After randomization, 20 of 30 children with storage phase dysfunction those were unresponsive or noncompliant to medical treatment received TPTNS treatment (test group) and 10 children underwent TPTNS with no current (sham group) for 12 weeks. Fourteen healthy children constituted the nonsymptomatic group. UB levels, dysfunctional voiding and incontinence scoring system (DVISS), voiding diary, and quality of life (QoL) scores were assessed before and after treatment in the treatment groups. QoL scores, overall and day-time DVISS scores were significantly decreased in both sham and test groups (p < 0.05). In addition to these findings, the frequency of incontinence and urgency episodes were also significantly reduced (p < 0.05) in the TPTNS treatment group. This effect in the test group was still valid 2 years after intervention. There was no significant difference in UBs measurements between treatment and nonsymptomatic groups and between pretreatment and posttreatment measurements of test and sham groups. TPTNS is an efficient minimally invasive treatment in children with FVD who do not respond to medical treatment. TPTNS provides a significant improvement on episodes of frequency, episodes of incontinence, overall and day-time DVISS scores, and QoL scores. The effectiveness of treatment continues even at the end of the second year of intervention. UBs were not found to be predictive in terms of diagnosis and evaluating the treatment response.


Evaluating the refractive index, thickness and porosity of ultrathin cellulose nanocrystal films with different polymorphs by SPR technique.
It is of crucial importance to know the quality of ultrathin films deposited on surface plasmon resonance (SPR) sensors prior to adsorption experiments. In this study, the optical properties of ultrathin cellulose nanocrystal films with various polymorphs (cellulose I, cellulose II and the hybrid of cellulose I/II), which deposited on gold surface of SPR sensors, were determined by a two-medium SPR technique and the influences of the second medium were assessed as well. The measured refractive index for ultrathin cellulose nanocrystal films with polymorphs of cellulose I, I/II and II was 1.453, 1.462 and 1.464, respectively, with a low margin of error about 0.2%. The porosity of according CNC films on SPR sensors was assessed to be 20.8%, 19.0%, and 18.5%. The measured film thickness for all deposited cellulose nanocrystal films was in the range of 25-35 nm, with a margin of error about 5%, accorded well with that examined by quartz crystal microbalance. The results showed that SPR surveys combined with Winspall analysis allow for simultaneous determination of the thickness, refractive index and the derived porosity, and provide a facile in situ quality control for the modified SPR sensors prior to adsorption experiments.


Generation of solar spicules and subsequent atmospheric heating.
Spicules are rapidly evolving fine-scale jets of magnetized plasma in the solar chromosphere. It remains unclear how these prevalent jets originate from the solar surface and what role they play in heating the solar atmosphere. Using the Goode Solar Telescope at the Big Bear Solar Observatory, we observed spicules emerging within minutes of the appearance of opposite-polarity magnetic flux around dominant-polarity magnetic field concentrations. Data from the Solar Dynamics Observatory showed subsequent heating of the adjacent corona. The dynamic interaction of magnetic fields (likely due to magnetic reconnection) in the partially ionized lower solar atmosphere appears to generate these spicules and heat the upper solar atmosphere.


Treatment of small cerebral gliomas with CT-aided stereotaxic curietherapy.
In cases of stereotaxic irradiation of small tumors, CT makes possible a precise determination of tumor volume and dosage and permits exact positioning of the therapy probe. Iridium 192 wires are interstitially implanted in the tumor (mostly gliomas Grades I-III) after biopsy. The results with 98 CT-controlled patients (from a series of 213 implantations in midline tumors) have provided new information about the method of treating small cerebral gliomas.


Mutations obstructing ATP's emplacement in KIF2A nucleotide-binding pocket causes parenchymal malformations, motor developmental delay, with intellectual disability.
KIF2A-related tubulinopathy (MIM: #615411) is a very rare disorder that was clinically characterized as microcephaly, epilepsy, motor developmental disorder (MDD), and various malformations of cortical development, but intellectual disability (ID) or global developmental delay (GDD) was rarely reported in the patients. Quad whole-exome sequencing (WES) was performed on the proband, the older brother, and their parents. Sanger sequencing was used to verify the candidate gene variant. The proband, a 23-month-old boy, was previously diagnosed with GDD, and his brother, aged nine years, had ID; both were born to a healthy couple. Quad-WES detected a novel heterozygous KIF2A variant, c.1318G>A (p.G440R), in both the brothers but not in the parents. In-silico analysis revealed that the variants G440R and G318R (which were previously reported in the only reported patient with GDD) lead to markedly enlarged side chains and hinder ATP's emplacement in the NBD pocket. The type of KIF2A variants that sterically hinder ATP emplacing in KIF2A NBD pocket may be associated with the intellectual disability phenotype; however, further studies are needed. Findings in this case also suggest a rare parental germline mosaicism of KIF2A G440R.


The influence of bone mineral density and bisphosphonate therapy on the determinants of oral health and changes on dental panoramic radiographs in postmenopausal women.
The aim of this study was to assess the determinants of oral health including the number of decayed, missing, and filled teeth (DMFT) and periodontal indices in postmenopausal women with osteoporosis, osteoporosis treated with bisphosphonate therapy, and control group and to examine the correlation between dental panoramic indices (Mental Index-MI, Mandibular Cortical Index-MCI) and bone mineral density in these three groups of patients. The presented non-interventional study involved 120 postmenopausal women: women with osteoporosis (O) (n = 45), women with osteoporosis treated with bisphosphonates (OBP) (n = 45), and control group (C) (n = 30). DMFT, plaque, gingival and papilla bleeding index, pocket depth, clinical attachment loss, and the presence of periodontitis were evaluated for each patient. MI and MCI of all participants were measured on a dental panoramic radiograph. Group OBP showed significantly higher gingival, bleeding index and deeper pocket depth than C and/or O group. No significant differences were found in MI (p = .303) or MCI (p = .06) in all the examined groups. Also, there were no significant differences between the three groups in the presence of periodontitis as well as in the DMFT index. BP therapy could have a negative influence on periodontal health. Further, MI and MCI are not precise diagnostic tools for diagnosing low BMD in postmenopausal women. BP therapy could have a negative influence on the determinants of oral health in postmenopausal women with osteoporosis.


Poleward heat transport by the atmospheric heat engine.
The atmospheric heat transport on Earth from the Equator to the poles is largely carried out by the mid-latitude storms. However, there is no satisfactory theory to describe this fundamental feature of the Earth's climate. Previous studies have characterized the poleward heat transport as a diffusion by eddies of specified horizontal length and velocity scales, but there is little agreement as to what those scales should be. Here we propose instead to regard the baroclinic zone--the zone of strong temperature gradients and active eddies--as a heat engine which generates eddy kinetic energy by transporting heat from a warmer to a colder region. This view leads to a new velocity scale, which we have tested along with previously proposed length and velocity scales, using numerical climate simulations in which the eddy properties have been varied by changing forcing and boundary conditions. The experiments show that the eddy velocity varies in accordance with the new scale, while the size of the eddies varies with the well-known Rhines beta-scale. Our results not only give new insight into atmospheric eddy heat transport, but also allow simple estimates of the intensities of mid-latitude storms, which have hitherto only been possible with expensive general circulation models.


Design of a surrogate for evaluation of methods to predict bone bending stiffness.
The high incidence of osteoporosis and related fractures demands for the use and development of methods capable of detecting changes in bone mechanical properties. The most common clinical and laboratory methods used to detect changes in bone mechanical properties, such as stiffness, strength, or flexural rigidity, include: mechanical testing, medical imaging, medical image-based analytical calculations, and medical image-based finite element analysis. However, the innate complexity of bone makes validation of the results from each method difficult. The current study presents the design, fabrication, and functional testing of a bi-material and computed tomography scan compatible bone-surrogate which provides consistent reproducible mechanical properties for methodological evaluation of experimental, analytical, and computational bone bending stiffness prediction methods.


Variability and Comprehensiveness of North American Online Available Physical Therapy Protocols Following Hip Arthroscopy for Femoroacetabular Impingement and Labral Repair.
To assess comprehensiveness and variability of postoperative physical therapy protocols published online following hip arthroscopy for femoroacetabular impingement (FAI) and/or labral repair. Surgeons were identified by the International Society for Hip Arthroscopy "Find a Surgeon" feature in North America (http://www.isha.net/members/, search August 10, 2016). Exclusion criteria included nonsurgeons and protocols for conditions other than hip arthroscopy for FAI and/or labral tear. Protocols were identified by review of surgeons' personal and departmental websites and evaluated for postoperative restrictions, rehabilitation components, and the time points for ending restrictions and initiating activities. Of 111 surgeons available online, 31 (27.9%) had postoperative hip arthroscopy physical therapy protocols available online. Bracing was used in 54.8% (17/31) of protocols for median 2-week duration (range, 1-6 weeks). Most protocols specified the initial postoperative weight-bearing status (29/31, 93.5%), most frequently partial weight-bearing with 20 pounds foot flat (20/29, 69.0%). The duration of weight-bearing restriction was median 3 weeks (range, 2-6) for FAI and median 6 weeks (range, 3-8) for microfracture. The majority of protocols specified initial range of motion limitations (26/31, 83.9%) for median 3 weeks (range, 1.5-12). There was substantial variation in the rehabilitation activities and time points for initiating activities. Time to return to running was specified by 20/31 (64.5%) protocols at median 12 weeks (range, 6-19), and return to sport timing was specified by 13/31 (41.9%) protocols at median 15.5 weeks (range, 9-23). There is considerable variability in postoperative physical therapy protocols available online following hip arthroscopy for FAI, including postoperative restrictions, rehabilitation activities, and time points for activities. This information offers residents, fellows, and established hip arthroscopists a centralized comparison of publicly available physical therapy protocols following hip arthroscopy. Practicing arthroscopists might find this analysis useful to compare various therapy strategies to their own recommendations. The variability we report can also provide inspiration for future efficacy research toward a more standard rehabilitation.


Lysis of spontaneous murine breast tumors by human interleukin 2-stimulated syngeneic T-lymphocytes.
In vitro and in vivo cytotoxic assay systems show that primary explants of spontaneous mammary tumors from CD8F1 mice are susceptible to lysis by interleukin 2-stimulated, syngeneic T-lymphocytes.


Isolation and characterization of a gene for a ryanodine receptor/calcium release channel in Drosophila melanogaster.
The nucleotide sequence of a 25.7 kilobase Drosophila melanogaster genomic DNA segment containing a gene for a ryanodine receptor/calcium release channel homologue has been determined. Computer analysis and partial cDNA cloning revealed 26 exons comprising the protein-coding sequence in this gene. The predicted protein is homologous in amino acid sequence and shares characteristic structural features with the mammalian ryanodine receptors. In blot hybridization analysis, a approximately 16 kilobase RNA species was identified abundantly in a 6-12 h embryo as the transcript from this gene. In situ hybridization to polytene chromosomes indicated that this gene locates at band position 44F on the second chromosome.


Prevalence and length of recovery of pusher syndrome based on cerebral hemispheric lesion side in patients with acute stroke.
The aim of this study was to determine if side of cerebral hemisphere lesion affects the prevalence and time course of pushing behavior (PB) after stroke. A total of 1660 patients with acute stroke were investigated. PB was assessed using the standardized Scale for Contraversive Pushing. Risk ratios were used to evaluate the differences in the prevalence of PB between right cerebral hemisphere-damaged (RCD) and left cerebral hemisphere-damaged (LCD) patients. The differences in the time course among 35 (27 RCD and 8 LCD) patients were evaluated by analyzing Scale for Contraversive Pushing scores with the Kaplan-Meier method using a log-rank test. PB was observed in 156 (9.4%) patients. The prevalence of PB was significantly higher in RCD (97 of 556 [17.4%]) than in LCD (57 of 599 [9.5%]) patients; risk ratio was 1.83 (95% CI, 1.35-2.49). The log-rank test indicated that RCD patients exhibited a significantly slower recovery than LCD patients (P=0.027). The number of RCD patients who exhibited PB was higher than that of LCD patients. The duration of recovery from PB was longer in RCD patients than in LCD patients.


The defect seen in the phosphatidylinositol hydrolysis pathway in HIV-infected lymphocytes and lymphoblastoid cells is due to inhibition of the inositol 1,4,5-trisphosphate 1,3,4,5-tetrakisphosphate 5-phosphomonoesterase.
Lymphocytes infected in vivo with HIV or lymphoblastoid cells exposed in vitro to either HIV or its envelope glycoprotein (gp120) show a defect in inositol polyphosphate-mediated signal transduction together with an associated abnormality in intracellular calcium regulation. The defect in patients reverses after treatment with the anti-retroviral agent zidovudine (AZT). We present evidence that the defect is at the level of the Ins (1,3,4,5)P4 5-phosphomonoesterase (PME) in these cells and that, though elevation of the intracellular ATP level partially down-regulates the activity of this enzyme, such changes alone are unable to account for the complete inhibition seen in HIV-infected cells.


Novel properties of recombinant Sso7d-Taq DNA polymerase purified using aqueous two-phase extraction: Utilities of the enzyme in viral diagnosis.
Using Sso7d from Sulfolobus solfataricus as the DNA binding protein fused to Taq DNA polymerase at its amino terminus, we report the hyper-expression and a novel purification methodology of Sso7d-Taq polymerase (S-Taq) using aqueous two-phase extraction system followed by Ni-affinity chromatography. The utility of such a fusion enzyme in carrying out PCR of human genes from whole blood directly and in detecting hepatitis B virus from clinical samples is demonstrated in this article. We present data on the enhanced thermo-stability of S-Taq DNA polymerase over Taq DNA polymerase and also provide evidence of its higher stability with detergents in comparison to Taq polymerase. The purified S-Taq protein showed acceptable limits of host genomic DNA levels without the use of DNases and other DNA precipitating agents and shows promising potential for use in PCR based diagnostics, in-situ PCR's and forensic science.


[Contractile function of the heart and its reactivity to noradrenaline in the process of the bodies adaptation to moderate altitudes].
In acute experiments on rats adapted for 4 weeks to the altitude of 2100 m it was found that at the early period of altitude adaptation the left ventricular systolic pressure and the maximal rates of pressure rise and fall considerably exceeded the values of control tests conducted in Moscow. Later on the parameters of the cardiac contractile function stabilized gradually. In 4 weeks the heart of adapted animals showed a greater increment of the contractility parameters in response to noradrenaline injection and a higher stability of the contractile function at very high heart rates.


The prevalence and outcome of effusive constrictive pericarditis: a systematic review of the literature.
There is sparse information on the epidemiology of effusive constrictive pericarditis (ECP). The objective of this article was to review and summarise the literature on the prevalence and outcome of ECP, and identify gaps for further research. The prevalence of ECP ranged from 2.4 to 14.8%, with a weighted average of 4.5% [95% confidence interval (CI) 2.2-7.5%]. Sixty-five per cent (95% CI: 43-82%) of patients required pericardiectomy regardless of the aetiology. The combined death rate across the studies was 22% (95(CI: 4-50%). The prevalence of ECP is low in non-tuberculous pericarditis, while pericardiectomy rates are high and mortality is variable. In this review, of 10 patients identified with tuberculous ECP, only one presumed case had a definite diagnosis of ECP. Appropriate studies are needed to determine the epidemiology of ECP in tuberculous pericarditis, which is one of the leading causes of pericardial disease in the world.


Percutaneous Portal Vein Embolization Using a Simplified Sheathless 18-Gauge Trocar Needle Approach: Review of Efficacy and Safety.
Portal vein (PV) embolization (PVE) is traditionally performed via a PV sheath with selective embolization of PV branches. Here, the efficacy and safety of PVE with the use of only an 18-gauge needle is reported. Consecutive patients who underwent PVE from 2009 through 2017 were retrospectively reviewed. Forty-five patients (mean age, 60 y ± 7.6; 38 men) underwent 45 PVE procedures. Hepatocellular carcinoma, cholangiocarcinoma, and metastases accounted for 26 (58%), 13 (29%), and 6 (13%) patients, respectively. PVE was performed by puncturing a branch of right PV with an 18-gauge needle under US guidance. Via the same needle, direct portography was performed, followed by PVE with an N-butyl cyanoacrylate/Lipiodol mixture. Percentage increase of future liver remnant (FLR) volume and increase in ratio of FLR to total liver volume were estimated as measures of efficacy. Complications were reported according to Society of Interventional Radiology classification. Fluoroscopy time, procedure time, and dose-area product (DAP) were recorded. Technical success rate was 100%. The median DAP, fluoroscopy time, and procedure time were 74,387 mGy·cm2 (IQR, 90,349 mGy·cm2), 3.5 min (IQR, 2.10 min), and 24 min (IQR, 10.5 min). Among the 23 patients with complete CT volumetry data, mean increase in the ratio of FLR to total liver volume and percentage increase of FLR volume were 12.5% ± 7.7 and 50% ± 33, respectively. There were 3 minor complications (asymptomatic nonocclusive emboli in FLR) and 3 major complications (1 hepatic vein emboli, 1 subphrenic collection, and 1 hepatic infarct). PVE via a sheathless 18-gauge needle approach is feasible, with satisfactory FLR hypertrophy.


Crystal structure of the N-terminal domain of E. coli Lon protease.
We report here the first crystal structure of the N-terminal domain of an A-type Lon protease. Lon proteases are ubiquitous, multidomain, ATP-dependent enzymes with both highly specific and non-specific protein binding, unfolding, and degrading activities. We expressed and purified a stable, monomeric 119-amino acid N-terminal subdomain of the Escherichia coli A-type Lon protease and determined its crystal structure at 2.03 A (Protein Data Bank [PDB] code 2ANE). The structure was solved in two crystal forms, yielding 14 independent views. The domain exhibits a unique fold consisting primarily of three twisted beta-sheets and a single long alpha-helix. Analysis of recent PDB depositions identified a similar fold in BPP1347 (PDB code 1ZBO), a 203-amino acid protein of unknown function from Bordetella parapertussis, crystallized as part of a structural genomics effort. BPP1347 shares sequence homology with Lon N-domains and with a family of other independently expressed proteins of unknown functions. We postulate that, as is the case in Lon proteases, this structural domain represents a general protein and polypeptide interaction domain.


[Transduction of oncogene by retrovirus: structural alternation of a foreign DNA inserted into retrovirus].
Frequent modification such as a point mutation or a deletion of a DNA inserted into retrovirus DNA vector is observed. Activation of onco gene in retroviral genome might be caused by such mechanisms.


Adverse childhood experiences (ACEs) and medically traumatic events (TEs) in adolescents and young adults (AYAs) with cancer: a report from the Promoting Resilience in Stress Management (PRISM) randomized controlled trial.
In adolescents and young adults (AYAs) with cancer, we examined (1) the distribution and type of traumatic events (TEs) experienced prior to baseline assessment and (2) how a resilience intervention, Promoting Resilience in Stress Management (PRISM), impacted changes in patient-reported outcomes (PROs) for AYAs with and without TEs. AYAs (12-25 years) within 1-10 weeks of diagnosis of new malignancy or ever diagnosed with advanced cancer were enrolled and randomly assigned to usual care (UC) with or without PRISM. To assess TEs, we screened medical records for traditionally defined adverse childhood experiences (ACEs) and medical traumatic events. Age-validated PROs assessed resilience, benefit-finding, hope, generic health-related quality of life (QoL), cancer-specific QoL, depression, and anxiety at enrollment and 6 months later. We calculated effect sizes (Cohen's d) for PRISM vs. UC effect on PRO score change at 6 months for 1+ TEs and 0 TE groups. Ninety-two AYAs enrolled and completed baseline surveys (44-UC, 48-PRISM; N = 74 at 6 months, 38-UC, 36-PRISM); 60% experienced 1+ TEs. PROs at baseline were similar across groups. PRISM's effect on score change was greater (Cohen's d ≥ 0.5) for the 1+ TE group on domains of benefit-finding and hope; and similar (d < 0.5) on domains of resilience, depression, anxiety, and both generic and cancer-specific QoL. In AYAs with cancer, TEs occurred at similar rates as the general population. PRISM may be particularly helpful for improving benefit-finding and hope for those who have experienced TEs.


A cantilevered swinglock removable partial denture design for the treatment of the partial mandibulectomy patient.
The use of the swinglock denture concept in the treatment of patients who have undergone mandibulectomy is discussed. The rationale for its use in cases with and without reconstruction is also discussed by reference to three treated cases.


Induction of the Adaptive Response in Mice Exposed to He-Ne Laser and X-Ray Radiation.
We studied the dose-dependent induction of in vivo adaptive response in the bone marrow and blood of mice exposed to low-intensity radiation of He-Ne laser (633 nm) and X-ray radiation by the severity of cytogenetic injury and intensity of ROS production, respectively. Induction of the adaptive response in mice preexposed to He-Ne laser and X-ray radiation depended on the adaptive dose and the interval between the adaptive and main doses and correlated with changes in ROS generation. The adaptive response after exposure to low-intensity ionizing and non-ionizing radiation was observed in the same dose range, which attests to similar mechanisms of its induction.


Stem Cell Enriched Fat Grafts versus Autologous Fat Grafts in Reconstructive Surgery: Systematic Review and Meta-Analysis.
To compare the outcomes of stem cell-enrichment fat grafting (SCEFG) versus autologous fat grafting (AFG) for reconstructive purposes. A systematic review and meta-analysis was performed as per the preferred reporting items for systematic reviews and meta-analyses. Guidelines and a search of electronic information was conducted to identify all Randomised Controlled Trials (RCTs), case-control studies and cohort studies comparing the outcomes of SCEFG versus AFG. Volume retention, fat necrosis, cancer recurrence, redness and swelling, infection, and cysts were primary outcome measures. Secondary outcome measures included patient satisfaction post-surgery, scar assessment, operation time and number of fat grafting sessions. Fixed and random effects modelling were used for the analysis. 16 studies enrolling 686 subjects were selected. Significant differences between the SCEFG and AFG groups were seen in mean volume retention (standardised mean difference = 3.00, P < 0.0001) and the incidence of redness and swelling (Odds Ratio [OR] = 441, P = 0.003). No significant difference between the two groups in terms of fat necrosis (OR = 2.23, P = 0.26), cancer recurrence (OR = 1.39, P = 0.58), infection (OR = 0.30, P = 0.48) and cysts (OR = 0.88, P = 0.91). For secondary outcomes, both cohorts had similar results in patient satisfaction, scar assessment and number of fat grafting sessions. Operation time was longer for the intervention group. SCEFG offers better outcomes when compared to AFG for reconstructive surgery as it improves the mean volume retention and does not worsen patient satisfaction and surgical complications except for self-limiting redness and swelling. Further clinical trials are recommended to support this argument and validate the use of SCEFG in clinical practice. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .


Comparison of pre- and intrapartum screening of group B streptococci and adherence to screening guidelines: a cohort study.
Implementation of guidelines for group B streptococcal (GBS) prepartum screening (PS) rarely has been prospectively evaluated. To assess PS at 35-37 weeks of gestation and compare its predictive value to that of an intrapartum screening (IS) within 7 days of delivery, a surveillance cohort study was conducted at a tertiary care center in Freiburg, Germany, during 2011-2012. Study participants included 937 pregnant women who had intrapartum cultures taken for vaginal and rectal GBS colonization. Colonization status was compared to PS, and intrapartum antibiotic prophylaxis (IAP) rates calculated. The neonates were tested for GBS transmission via cultures from their throats and external ear canals. While 67.5% (633/937) of study participants had a PS, only 22.7% (144/633) underwent a fully guideline-compatible PS. However, maternal GBS colonization rates were similar when comparing PS (18.5% [117/633]) versus IS (17.0% [133/784]). The positive predictive value of a positive PS result for GBS positivity at delivery was 77.2 %. Women with a positive PS received IAP in 89.3% of cases (75/84). The capsular serotype distribution pattern of colonizing GBS strains has not changed in comparison to our 2003-2004 study--one with a similar study design. Improved strategies for adoption of prepartum GBS screening are needed. • The prediction of prepartum GBS screening for intrapartum colonization status has not been well studied. • Longitudinal studies of GBS screening are needed for screening program evaluations and vaccine development. What is New: • The rate of GBS screening has improved over 10 years, and intrapartum GBS colonization prediction was accurate. • Serotype distribution was stable and suggests the potential long-term efficacy of GBS vaccines.


Occupational and ambient radiation exposures from Lu-177 DOTATATE during targeted therapy.
Lutetium-177 (DOTATATE) (177Lu; T1/2 6.7 days), a labelled β- and Auger-electron emitter, is widely used in treatment of neuroendocrine tumours. During performance of the procedure, staff and other patients can potentially receive significant doses in interception of the gamma emissions [113 keV (6.4%) and 208 keV (11%)] that are associated with the particle decays. While radiation protection and safety assessment are required in seeking to ensure practices comply with international guidelines, only limited published studies are available. The objectives of present study are to evaluate patient and occupational exposures, measuring ambient doses and estimating the radiation risk. The results, obtained from studies carried out in Riyadh over an 11 month period, at King Faisal Specialist Hospital and Research Center, concerned a total of 33 177Lu therapy patients. Patient exposures were estimated using a calibrated Victoreen 451P survey meter (Fluke Biomedical), for separations of 30 cm, 100 cm and 300 cm, also behind a bed shield that was used during hospitalization of the therapy patients. Occupational and ambient doses were also measured through use of calibrated thermoluminescent dosimeters and an automatic TLD reader (Harshaw 6600). The mean and range of administered activity (in MBq)) was 7115.2 ± 917.2 (4329-7955). The ambient dose at corridors outside of therapy isolation rooms was 1.2 mSv over the 11 month period, that at the nursing station was below the limit of detection and annual occupational doses were below the annual dose limit of 20 mSv. Special concern needs to be paid to comforters (carers) and family members during the early stage of radioisotope administration.


[Physiotherapy, exercise and strength training and physical therapies in the treatment of fibromyalgia syndrome].
A guideline for the treatment and diagnostic procedures for fibromyalgia syndrome (FMS) was developed in cooperation with 10 German medical and psychological associations and 2 patient self-help groups. A systematic literature search including all controlled studies evaluating physiotherapy, exercise and strength training as well as physical therapies was performed in the Cochrane Collaboration Reviews (1993-12/2006), Medline (1980-12/2006), PsychInfo (1966-12/2006) and Scopus (1980-12/ 2006). Levels of evidence were assigned according to the classification system of the Oxford Centre for Evidence-Based Medicine. Grading of the strengths of recommendations was done according to the German program for disease management guidelines. Standardized procedures to reach a consensus on recommendations were used. Aerobic exercise training is strongly recommended (grade A) and the temporary use of whole body hyperthermia, balneotherapy and spa therapy is recommended (grade B). The significance which can be assigned to most of the studies on the various procedures for therapy is restricted due to short study duration (mean 6-12 weeks) and small sample sizes.


Rapid Synthesis of Cryo-ET Data for Training Deep Learning Models.
Deep learning excels at cryo-tomographic image restoration and segmentation tasks but is hindered by a lack of training data. Here we introduce cryo-TomoSim (CTS), a MATLAB-based software package that builds coarse-grained models of macromolecular complexes embedded in vitreous ice and then simulates transmitted electron tilt series for tomographic reconstruction. We then demonstrate the effectiveness of these simulated datasets in training different deep learning models for use on real cryotomographic reconstructions. Computer-generated ground truth datasets provide the means for training models with voxel-level precision, allowing for unprecedented denoising and precise molecular segmentation of datasets. By modeling phenomena such as a three-dimensional contrast transfer function, probabilistic detection events, and radiation-induced damage, the simulated cryo-electron tomograms can cover a large range of imaging content and conditions to optimize training sets. When paired with small amounts of training data from real tomograms, networks become incredibly accurate at segmenting in situ macromolecular assemblies across a wide range of biological contexts.


A quantitative morphological study of the bovine vaginal epithelium during the oestrous cycle.
Samples were collected post mortem from 28 cycling parous cows. At all stages of the oestrous cycle, the cranial vaginal epithelium was thinner (P less than 0.001), had fewer cell layers (P less than 0.001) and proportionally more superficial columnar cells (P less than 0.001) than more caudal regions. At all stages of the cycle there were more intraepithelial lymphocytes caudally than in more cranial regions (P less than 0.001). At oestrus the proportion of superficial columnar cells was maximal, having increased from the late dioestrus proportions of 1, 29 and 40 per cent of all superficial cells to 15, 54 and 83 per cent in caudal, middle and cranial regions, respectively (P less than 0.02). However, the epithelial thickness in all regions of the vagina was greatest at metoestrus (P less than 0.02). At this stage, the number of cell layers was greater than at any other stage (P less than 0.05).


Actinomycin synthetases. Multifunctional enzymes responsible for the synthesis of the peptide chains of actinomycin.
Two enzymes were purified from actinomycin-synthesizing Streptomyces chrysomallus which could be identified as peptide synthetases involved in the biosynthesis of actinomycin. Actinomycin synthetase II activates the first two amino acids of the peptide chains of the peptide lactone antibiotic, threonine and valine (or isoleucine), as thioesters via their corresponding adenylates. It is a single polypeptide chain of Mr 225,000. Similarly, actinomycin synthetase III activates proline, glycine, and valine (the remaining three amino acids in the antibiotic) as thioesters and is a single polypeptide chain of about Mr 280,000. It also carries the methyltransferase function(s) for N-methylation of thioesterified glycine and valine. In addition, it catalyzes the formation of cyclo(sarcosyl-N-methyl-L-valine) from glycine, L-valine, and S-adenosyl-L-methionine at the expense of ATP. Although the cell-free synthesis of the peptide lactone was not as yet accomplished, the data provide evidence that together with the 4-methyl-3-hydroxyanthranilic acid-activating enzyme (now designated as actinomycin synthetase I) all amino acid-activating protein components of the actinomycin-synthesizing enzyme complex are identified.


Administration of myelin basic protein-coupled spleen cells prevents experimental allergic encephalitis.
Intravenous administration of mouse myelin basic protein covalently coupled with chromic chloride to syngeneic spleen cells (MBP-SC) prevents the subsequent induction of experimental allergic encephalitis (EAE). Whereas 1 in 28 mice receiving MBP-SC developed EAE after immunization with mouse spinal cord homogenate (MSCH) and adjuvants, 16 out of 25 mice receiving ovalbumin-coupled spleen cells (OA-SC) had EAE following encephalitogenic challenge. The effect of administration of antigen-coupled spleen cells on in vitro proliferation responses is shown.


Polymeric microspheres as drug carriers.
The use of polymeric microsphere systems (including polypeptides) as vehicles for delivering drugs by a variety of routes is considered with particular reference to parenteral administration. Microsphere formulation is discussed with emphasis on the role of surface properties and how these can determine the fate of administered particles. Studies on the use of adsorbed block copolymers that can alter processes of protein uptake onto particle surfaces (opsonization) as well as the interaction of particles with macrophages (cell-particle adhesion) are described. Data are presented which show that by the selection of the appropriate coating material it is possible to direct particles within the body to sites such as the lung, liver, bone marrow, or to retain materials within the systemic circulation.


A low-cost easily implementable physiotherapy intervention clinically improves gait implying better adaptation to lower limb prosthesis: a randomized clinical trial.
Lower limb amputation highly impacts the lives of individuals. The inability to walk due to difficulties in adapting to wearing prosthesis can potentially result in physical degeneration and comorbidity in this population. In this randomized clinical trial study, we investigated if a low-cost and easily implementable physiotherapy intervention was effective in improving gait performance and adaptation to lower limb prosthesis in individuals with an amputation. A total of 26 individuals participated in the study, 16 with lower limb amputation and 10 without amputation. Participants with amputation were further divided in intervention and control groups. The intervention group underwent a rehabilitation protocol aimed at strengthening muscles and improving prosthesis adaptation. Muscle strengthening targeted the hip segment, prioritizing the abdominal muscles, hip flexors, extensors, adductors and abductors, followed by cicatricial mobilization and weight-bearing on the stump for desensitization. Assessment and measures were performed across the kinetic and kinematic parameters of gait. In the comparison between pre-and post-intervention, a significant increase in gait speed (0.68-2.98, 95% CI, 1.83, effect size ES) and cadence (0.56-2.69, 95% CI, 1.63, ES) was found between groups and time points. Step (0.73-3.11, 95% CI, 1.92, ES) and stride length (0.62-2.84, 95% CI, 1.73) increased between pre- and post-intervention, while in the control group both variables remained smaller. The intervention group decreased stance phase as a percentage of gait cycle between pre- and post-intervention (- 1.33-0.62, 95% CI, - 36, ES), while it increased in the control group. Improvement in a combination of important gait parameters indicates that the intervention protocol promoted the adaptation to prosthesis and the functional independence of individuals with lower limb amputation. It is recommended that the participants continue receiving follow-up assessments and rehabilitation interventions.


Linking DNA Damage and Hormone Signaling Pathways in Cancer.
DNA damage response and repair (DDR) is a tightly controlled process that serves as a barrier to tumorigenesis. Consequently, DDR is frequently altered in human malignancy, and can be exploited for therapeutic gain either through molecularly targeted therapies or as a consequence of therapeutic agents that induce genotoxic stress. In select tumor types, steroid hormones and cognate receptors serve as major drivers of tumor development/progression, and as such are frequently targets of therapeutic intervention. Recent evidence suggests that the existence of crosstalk mechanisms linking the DDR machinery and hormone signaling pathways cooperate to influence both cancer progression and therapeutic response. These underlying mechanisms and their implications for cancer management will be discussed.


Pathophysiology of multiple sclerosis damage and repair: Linking cerebral hypoperfusion to the development of irreversible tissue loss in multiple sclerosis using magnetic resonance imaging.
Reduced cerebral perfusion has been observed in multiple sclerosis (MS) and may contribute to tissue loss both acutely and chronically. Here, we test the hypothesis that hypoperfusion occurs in MS and relates to the presence of irreversible tissue damage. In 91 patients with relapsing MS and 26 healthy controls (HC), gray matter (GM) cerebral blood flow (CBF) was assessed using pulsed arterial spin labeling. GM volume, T1 hypointense and T2 hyperintense lesion volumes (T1LV and T2LV, respectively), and the proportion of T2-hyperintense lesion volume that appears hypointense on T1-weighted magnetic resonance imaging (T1LV/T2LV) were quantified. GM CBF and GM volume were evaluated globally, as well as regionally, using an atlas-based approach. Global GM CBF was lower in patients (56.9 ± 12.3 mL/100 g/min) than in HC (67.7 ± 10.0 mL/100 g/min; p < 0.001), a difference that was widespread across brain regions. Although total GM volume was comparable between groups, significant reductions were observed in a subset of subcortical structures. GM CBF negatively correlated with T1LV (r = -0.43, p = 0.0002) and T1LV/T2LV (r = -0.37, p = 0.0004), but not with T2LV. GM hypoperfusion occurs in MS and is associated with irreversible white matter damage, thus suggesting that cerebral hypoperfusion may actively contribute and possibly precede neurodegeneration by hampering tissue repair abilities in MS.